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<SOH> SUMMARY OF THE INVENTION <EOH>The inventors have found that notochord enriched media and/or factors derived therefrom, stimulate proteoglycan production in bovine disc chondrocytes. As well, the inventors have verified the expression of several genes important to chondrocyte metabolism that are increased in expression as a result of culture with these factors. These genes have been shown to be active in bovine and human disc chondrocytes. Therefore, the present invention relates to a composition for enhancing the production of proteoglycan in a cell or animal in need thereof comprising notochord enriched media and/or one or more factors derived from notochord enriched media. Preferably the notochord enriched media, and/or one or more factors derived therefrom, are from a nonchondrodystrophic animal, for example, nonchondrodystrophic canines, rabbits or felines, and the cell is a chondrocytic cell, such as a chondrocyte from the disc or articula cartilage. The present invention further involves a method for enhancing proteoglycan production comprising administering to a cell or animal in need of such treatment, an effective amount of a composition comprising notochord enriched media and/or one or more factors derived therefrom. The invention also relates to a use of a composition comprising notochord enriched media and/or one or more factors derived therefrom to enhance the production of proteoglycan in a cell or animal in need thereof, and a use of a composition comprising notochord enriched media and/or one or more factors derived therefrom to prepare a medicament to enhance the production of proteoglycan in a cell or animal in need thereof. Preferably the cell is a chondrocyte and the subject is a mammal, in particular, humans. In an embodiment of the present invention, the cell is an intervertebral chondrocyte, therefore there is provided a method of treating degenerative disc disease comprising administering to a cell or animal in need of such treatment, an effective amount of a composition comprising notochord enriched media and/or one or more factors derived therefrom. The invention also relates to a use of a composition comprising notochord enriched media and/or one or more factors derived therefrom to treat degenerative disc disease in a cell or animal in need thereof, and a use of a composition comprising notochord enriched media and/or one or more factors derived therefrom to prepare a medicament to treat degenerative disc disease in a cell or animal in need thereof. The present invention further relates to a pharmaceutical composition for enhancing the production of proteoglycan comprising notochord enriched media and/or one or more factors derived therefrom and a pharmaceutically acceptable carrier. The present invention further relates to a method of preparing notochord enriched media comprising: (a) separating a nucleus pulposus from an intervertabral disc of a nonchondrodystrophic animal to provide a total nucleus digest; (b) separating notochord cells from the total nucleus digest; and (c) purifying the notochord cells and culturing the notochord cells in a media to provide notochord enriched media. Other features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples while indicating preferred embodiments of the invention are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. |
High frequency integrated circuit (hfic) microsystems assembly and method for fabricating the same |
High frequency integrated circuit (HFIC) microsystems assembly and method for fabricating the same are disclosed. Presented HFIC assembly method has the optimized structure for minimizing the losses in transmitting electronic and electromagnetic energy in interconnects; it optimizes the area used for interconnects and eliminates most hazardous materials from the assembly process making it an environmentally friendly alternative for IC assembly purposes. This versatile assembly process was developed specifically for HFIC packaging, but its versatility expands its usage from monolithic microwave integrated circuit (MMIC) packaging to partial PCB assemblies and due to environmental friendliness potentially replacing other PCB techniques especially in high performance applications. HFIC assembly comprises a first substrate (702, 703) and a second substrate (701) of conductor-on-insulator or similar having high aspect ratio trenches and conductors (705, 706, 707, 708) as well as a chip therebetween. A common ground (707, 708, 710, 710′, 711, 711′) formed by the first and second substrates encompass the chip at least adjacent the HF-signal paths (706). |
1. Large area planar high frequency integrated circuit (HFIC) assembly comprising: a first substrate (702, 703) having at least one recess for a chip (709) having pads and a chip face and at least one planar surface (710, 711′) on the side exposing chip pads and the said substrate being conductive at least at this functional side; a second substrate (701) of conductor-on-insulator having high aspect ratio trenches and conductors (705, 706, 707, 708) comprising planar surfaces (708, 710′), grounding and signal paths for HF- and DC-signals on a functional side arranged to provide connections with chip pads (612), points of peripheral contacts and area for common ground, when assembled in aligned relation to the said planar surfaces (710, 711′, 708, 710′) of each functional side of the first and second substrates, and the said signal paths and grounding each having a conductor facing the functional side of the first substrate, and the said high aspect ratio conductors increasing the air gap (616) at the chip face; common ground areas (710, 710′) between the first and second substrates and these being attached to one substrate and forming shielded cavities (615) for the signal paths (705, 706), and a common ground (707, 708, 710, 710′, 711, 711′) formed by the first and second substrates encompassing the chip at least adjacent to the HF-signal paths (706); connections between each pad (610, 612) of the chip and the respective signal path (603, 611) of the second substrate. 2. The HFIC assembly according to claim 1, further comprising: a metallized semiconductor, e.g., silicon, as the first substrate. 3. The HFIC assembly according to claim 1, further comprising: a metal structure, e.g., Ni, Cu, Ag, Au as the first substrate. 4. The HFIC assembly according to claim 3, further comprising: the first substrate having a core metal and the metallized layer being more conductive than the core metal. 5. The HFIC assembly according to claim 1, further comprising: a molded conductive or metallized non-conductive polymer as the first substrate. 6. The HFIC assembly according to any of claims 1 to 5, further comprising: a metallized semiconductor-on-insulator as the second substrate. 7. The HFIC assembly according to any of claims 1 to 5, further comprising: a metal-on-insulator (MOI), e.g., Ni, Cu, Ag, Au on insulator as the second substrate. 8. The HFIC assembly according to any of claims 1 to 5, further comprising: a freestanding metal as the second substrate. 9. The HFIC assembly according to claim 6 or 7, further comprising: a low dielectric constant material as the insulator. 10. The HFIC assembly according to claim 6 or 7, further comprising: a high dielectric constant material as the insulator. 11. The HFIC assembly according to any of claims 1 to 10, further comprising: the conductors at a height of 20-3000 um. 12. The HFIC assembly according to any of claims 1 to 11, further comprising: an inverted trapped dual mode structure as the HF-signal path, wherein a trapped coplanar waveguide (CPW) is at the chip end and an inverted microstrip line at the opposite end in order to match the pad width of the chip with an essentially greater width of the point of contact at the opposite end. 13. The HFIC assembly according to any of claims 1 to 11, further comprising: an inverted trapped dual mode structure as the HF-signal path, wherein a trapped coplanar waveguide (CPW) is at the chip end and an trapped inverted microstrip line at the opposite end in order to match the pad width of the chip with essentially greater width of the point of contact in the opposite end. 14. The HFIC assembly according to any of claims 1 to 11, further comprising: an inverted trapped structure as the HF-signal path, wherein a trapped coplanar waveguide (CPW) is formed both at the chip end and at the opposite end in order to match the pad width of the chip with an essentially greater width of the point of contact at the opposite end. 15. The HFIC assembly according to any of claims 1 to 14, wherein the HFIC assembly is applicable to at least one of the following embodiments: low frequency circuit, MEMS component, optoelectronic integrated circuit, optical transmission line or optical fiber assembly. 16. The HFIC assembly according to any of claims 1 to 14, further comprising: a case with a plurality of connectors with its periphery and each connector connected to respective point of contact of the second substrate inside the case. 17. The HFIC assembly according to any of claims 1 to 15, wherein: the HFIC assembly forms a subassembly embedded in a printed circuit board (PCB). 18. The HFIC assembly according to any of claims 1 to 15, wherein the HFIC assembly is used as a stand alone PCB board. 19. Fabrication method for a HFIC-assembly having a self supporting integral carrier structure and at least one HFIC-chip comprising: making the first conductive substrate of HFIC assembly having at least one chip recess; making the second substrate of HFIC assembly with a semiconductor-on-insulator (SOI) wafer; installing a HFIC chip on the second substrate; bonding of the first and second substrates. 20. The fabrication method according to claim 19 further comprising: the first conductive substrate is made of a semiconductor wafer, which is first patterned and etched, then coated with a metal layer. 21. The fabrication method according to claim 19 or 20 further comprising: the semiconductor-on-insulator wafer is first patterned and etched to form high aspect ratio conductors having face and side surfaces as well as bottom surfaces between them. 22. The fabrication method according to claim 21 further comprising: at least face and side surfaces are coated with a metal layer. 23. The fabrication method according to claim 22 further comprising: the bottom surfaces are also coated with a metal layer. 24. Fabrication method for a HFIC-assembly having a self supporting integral carrier structure and at least one HFIC-chip comprising: making the first conductive substrate of HFIC assembly having at least one chip recess; making the second substrate of HFIC assembly with a metal-on-insulator carrier; installing a HFIC chip on the second substrate; bonding of the first and second substrates. 25. The fabrication method according to claim 24 further comprising: the first substrate is a metal carrier or a metal carrier on insulator. 26. The fabrication method according to claim 24 further comprising: the insulator is used and metal structures are selectively deposited on the insulator forming high aspect ratio conductors. 27. The fabrication method according to claim 24 further comprising: the sacrifacial carrier is used and metal structures are selectively deposited forming high aspect ratio conductors and a supporting frame after which the sacrifacial carrier is removed and the frame is removed after the assembling of the assembly. 28. The fabrication method according to any of claims 25 to 27 further comprising: the metal structures are polished by chemical-mechanical-polishing (CMP) process. 29. Fabrication method for a HFIC-assembly having a self supporting integral carrier structure and at least one HFIC-chip comprising: making the first conductive substrate of HFIC assembly having at least one chip recess; making the second substrate of HFIC assembly with a polymer-on-insulator carrier; installing a HFIC chip on the second substrate; bonding of the first and second substrates. 30. The fabrication method according to claim 29 further comprising: making the first substrate of HFIC assembly with thermal and mechanical molding of conductive or non-conductive polymers-on-insulator. 31. The fabrication method according to claim 29 or 30 further comprising: making the second substrate of HFIC assembly with thermal and mechanical moldering, as well as etching of conductive and non-conductive polymers-on-insulator. 32. The fabrication method according to claim 30 or 31 further comprising: a metal layer coating is to be deposited on non-conductive polymer. 33. The fabrication method according to any of claims 29 to 31 further comprising: a metal layer coating is used for conductive polymer to improve the conductivity at the surface. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The way microwave circuits, patterned metal traces on various microwave substrates, have commercially been packaged for over 30 years utilizing test fixtures and metal housings have to large extend remained the same. Conventional electronic packaging has served the purpose of protecting electronic circuitry in low frequency applications while the package itself is the main cause of degradation in microwave applications. Today, group IV circuits have reached ULSI era and group III-V ICs LSI/VLSI era. Early monolithic ICs brought about the requirement to package ICs in single chip packages while MCMs become common in early 1990's. Today packaging methods are very diversified and often the infrastructure is complex. Until recently, EMS providers or packaging foundaries have packaged the ICs, while semiconductor industries have been providing the chips fabricated in clean rooms, where handling of PCBs is difficult. Feature sizes on PCBs have now become many orders of magnitude larger than feature sizes on chips. Thereby, conventional partitioning of electronic packaging has presented a clear conflict in further miniaturiazition of HFICs that calls for improved interface between the micro- and macroworlds. Traditionally common ground on HFIC chip had to be connected to the common ground of the substrate and metal housing through via holes, which tend to be large in size. Advangement in manufacturing technologies has reduced series inductance of via holes. Ideally one wants to connect the common ground on chip directly to the common ground of the assembly without via holes. However, this ideal option has not been presented in the market. Also, typically common ground of HFICs and low frequency ICs has been defined at different potential, making the integration of various ICs in one assembly more difficult. High frequency integrated circuit (HFIC) packaging has not yet reached the level where monolithic microwave ICs (MMICs) together with low and medium frequency ICs are integrated in a true 3D manner. Drayton et. al., U.S. Pat. No. 5,913,134, Jun. 15, 1999, discuss how passive MICs are created using Si micromachining. These types of circuits can easily be created by using isotropical silicon wet eching, e.g., KOH, 90 degree angle is not maintained and is clearly indicated by drawings. Drayton et. al. work is not suitable for integration of HFICs due to the fact that ICs typically have a large number of points of contacts and thus the substrate structure must become as compact as possible which is not attainable by Si wet etching. In HFICs, HF-signal is typically taken out from the chip differentially. In addition to ground and transmission lines one has to provide power and additionally, e.g., distribution of control signals, devided power and ground planes. Thus, a new type of HFIC microsystems assembly must represent is a clear extension to the formation of HFIC circuitry on chip. Typical MIC layouts are inherently simple in structure while highly integrated compact MMICs have complex structures. Thereby, Drayton's approach is not applicable. Lacking third signal plane and tightly held, large number of IC pads makes Drayton's approach void in this invention. Problems specifically related to HFIC assembly and generic technology in this field is discussed in the following publication: “High Frequency MultiChip Modules—Materials, Design and Fabrication Techniques”, Tarja A. Juhola, Royal Institute of Technology, May 2000, ISRN KTH/MVT/FR-00/1-SE, ISSN 0348-4467, TRITA-MVT Report 2000:1. |
<SOH> SUMMARY OF THE INVENTION <EOH>It is therefor an object of the present invention to provide high frequency integrated circuit (HFIC) Microsystems assembly and methods for fabricating the same. Fabrication-wise fully planar approach creates an optimal HFIC-assembly that can be manufactured cost-effectively with minimal transmission line lossies. The approach makes further miniaturiazition of ICs possible: it improves interface between micro- and macroworlds by enabling remarkable reduction of chip pads in size. The approach enables large area decoupling effects of power and ground on ICs to be transferred onto the assembly substrate. The invention optimizes the area used for interconnects. The method can easily be integrated with IC-processing and make the assembly design process more robust and reliable. Trapped CPW transmission lines used at the chip end enable integration of both group III-V and group IV based circuits in the same assembly. This invention eliminates most hazardous materials from the assembly making it an environmentally friendly alternative for existing PCB-technologies. This assembly process was aimed specifically for HFIC packaging, but its versatility expands the usage from monolithic microwave integrated circuit (MMIC) packaging to partial PCB assemblies and due to environmental friendliness potentially replacing other PCB techniques especially in high performance applications. In a preferred method of manufacturing the HFIC-assembly, a silicon-on-insulator wafer is used, being suitable for prototyping and small scale production. On the other hand, electroforming in particular, extends the scope of this invention to mass production, being cost effective manufacturing alternative for low frequency systems as well. All partitioned manufacturing stages: IC-fabrication, electronic packaging and PCB-assembly, can be done in an almost particle free clean room to avoid contamination from the outside and a process mismatch of incompatible materials. Due to the fact that ICs oftentimes have order of magnitude larger number of of transistors, and much larger number of I/Os in comparison to passive MICs structures (Drayton, et. al.), calls for special assembly approach, e.g., anisotropical Si micromachining. Only DRIE of the semiconductor will be able to create non-deformed irregular angles seen from the top creating shortest possible signal paths to the edge of the silicon substrate. Fully planar manufacturing enable chip-to-substrate transition without the use of wirebonding or any type of flipchip bumps or similar materials. This invention has the advantage of integral passive component integration, through postprocessing of ICs and/or embedding filters, inductors, etc. within the assembly. Postprocessed ICs are mounted in the same manner as non-postprocessed ICs. Preferably hermetic cover is used due to the fact that moisture absorbtion may be a problem at higher frequencies (in 10's of GHz) but even in a few GHz range. The proposed HFIC assembly allows effective hermetic sealing and minimizes electrical discontinuities and transmission losses. The characteristic features of the HFIC assembly according to the invention are presented in the accompanying claim 1 . The preferred methods of making the same are presented in claims 19 , 24 and 29 . Further objects, features advantages of the invention will become apparent from a consideration of the following description and the appended claims when taken in connection with the acconpanying drawings. |
Broadband communications |
Disclosed is a provisioning tool and method for a communications network, particularly a broadband communications network which is arranged to provision a service for a user based on information available via another user connection; this facilitates provision. In a preferred embodiment, a user device address is obtained from a user IP address. Provisioning and other network tools with additional or alternative advantageous features are disclosed and the provisioning tools disclosed may communicate with a variety of other tools including planning and inventory applications and agents associated with equipment, all of which are also independently provided, enabling provision of a variety of novel services and features dealing with a spectrum of problems arising in a communications environment, particularly a broadband environment. |
1-110. cancelled. 111. A method of provisioning a desired communications service for a user across a network, the method comprising obtaining information for a desired communications service to be provisioned based on information associated with an existing connection for the user and provisioning the desired communications service based on the information obtained. 112. A method according to claim 111, wherein the information for the desired communication service comprises an address for a network access device. 113. A method according to claim 112, wherein the information associated with an existing connection is based on a communications protocol address of a user device. 114. A method according to claim 113 wherein the communications protocol is the Internet Protocol. 115. A method according to claim 111 wherein the network access device is a cable modem and the address is a MAC address. 116. A method according to claim 111 further comprising providing a user interface for communicating user information. 117. A method according to claim 116 wherein the user interface comprises a provisioning interface for receiving a provisioning instruction from a provisioning operator. 118. A method according to claim 112 wherein the device address is obtained using a query expressed in a configuration protocol. 119. A method according to claim 112 wherein the configuration protocol is a Dynamic Host Configuration Protocol and the query is expressed as a Dynamic Host. Configuration Protocol Lease Query Message. 120. A method according to claim 116 wherein the user interface comprises a planning interface, the method comprising receiving user information comprising a desired service or a set of services for a plurality of users. 121. A method according to claim 116 wherein the interface comprises a customer front end interface for receiving user information from a user, preferably wherein the method comprises re-provisioning a service based on user authentication information and/or user information requesting a service. 122. A method according to claim 111 wherein the information obtained is an access device hardware identifier and wherein the method comprises obtaining the hardware address based on an existing IP address of the user and provisioning the communications service based on the hardware address obtained. 123. A method according to claim 111 including checking for available capacity prior to provisioning a service. 124. A method according to claim 111 including reporting provisioning of a service to a capacity management application after provisioning. 125. A method according to claim 111 further comprising receiving data identifying fraudulent use of a service, identifying an access device relevant to said fraudulent use and changing conditions of service provision via the access device. 126. A method according to claim 125 wherein said changing conditions of service provision comprises withdrawal of one or more services via the access device. 127. A method according to claim 111 wherein provisioning a service is effected by executing an activation script defining a workflow sequence and including at least one activation action. 128. A provisioning application having means for provisioning a broadband service to a user's termination device, based on an access device hardware identifier, further comprising by means for obtaining the access device hardware identifier based on a DHCP lease query message submitted to a DHCP server and containing an IP address of the user's termination device. 129. A service provisioning system, for provisioning communications services to user devices across a network, said system comprising means for receiving a user request for provision of a service via the network for use at a user termination device, which user termination device has a network protocol address and is connected to the network via an access device, said service provisioning system further comprising means to formulate a query containing said network protocol address and sending it to a configuration server for the network, means for receiving back from the configuration server a response to the query, said response comprising a hardware address for said access device, and means to provision the service in respect of the hardware address for the access device. 130. A computer readable medium containing instructions for performing a method according to claim 111. |
<SOH> BRIEF DESCRIPTION OF DRAWINGS <EOH>The provisioning of services in a communications network will now be described further, by way of example only, with reference to the accompanying drawings in which: FIG. 1 is a schematic diagram of a network environment for provisioning apparatus in accordance with one example of the system and methods herein described; FIG. 2 is a schematic diagram of message flow in provisioning apparatus carrying out provisioning according to one example of the systems and methods herein described; FIG. 3 is a schematic diagram of an architecture for a service management system using the provisioning apparatus of FIGS. 1 and 2 ; FIG. 4 is a schematic diagram of message flow in provisioning apparatus carrying out provisioning according to a further example of the systems and methods herein described; FIG. 5 is a schematic diagram of an initial set-up of a network environment for provisioning apparatus in accordance with a further example of the system and methods herein described; FIG. 6 is a schematic diagram of a network environment for provisioning apparatus in accordance with the example illustrated in FIG. 5 after a new customer edge router has been added to the network according to an example process; FIG. 7 is a schematic diagram of a network environment for provisioning apparatus in accordance with the example illustrated in FIG. 5 or 6 after a new customer edge router has been added to the network according to a further example process; FIG. 8 is a schematic diagram of a network environment for provisioning apparatus in accordance with the example illustrated in FIG. 5, 6 or 7 after a new customer edge router has been added to the network according to a further example process; FIG. 9 is a schematic diagram of an initial topology of the network environment for provisioning apparatus, before modification has taken place, according to an example of the system and methods herein described; FIG. 10 is a schematic diagram of a final topology of the network environment for provisioning apparatus, after modification has taken place, according to an example of the system and methods herein described; FIG. 11 is a schematic diagram of how the Set-Top Box Manager application may be incorporated into a network environment for provisioning apparatus according to one embodiment of the systems and methods herein described; FIG. 12 is a schematic diagram of an example of a Set-Top Box Manager screen display according to one embodiment of the systems and methods herein described; FIG. 13 is a schematic diagram of an example of a further Set-Top Box Manager screen display according to one embodiment of the systems and methods herein described; FIG. 14 is a schematic diagram of an example of a further Set-Top Box Manager screen display, which may be used to show ‘Audit’ information, according to one embodiment of the systems and methods herein described; FIG. 15 is a schematic diagram of a ‘Connection Window’ screen display according to one embodiment of the systems and methods herein described; FIG. 16 is a schematic diagram of an ‘Install’ Results List Panel screen display according to one embodiment of the systems and methods herein described; FIG. 17 is a schematic diagram of a ‘Forward Path’ Results List Panel screen display according to one embodiment of the systems and methods herein described; FIG. 18 is a schematic diagram of a ‘Reverse Path’ Results List Panel screen display according to one embodiment of the systems and methods herein described; FIG. 19 is a schematic diagram of a ‘Resources’ Results List Panel screen display according to one embodiment of the systems and methods herein described; FIG. 20 is a schematic diagram of an example of a further Set-Top Box Manager screen display, which may be used to show ‘Audit’ information, according to one embodiment of the systems and methods herein described; FIG. 21 is a schematic diagram of a Set-Top Box Manager Top Bar according to one embodiment of the systems and methods herein described; FIG. 22 is a schematic diagram of a one embodiment of the architecture of the Set-Top Box Manager, FIG. 23 is a schematic diagram of a Cable Modem Manager integrated into a distributed system according to one embodiment of the systems and methods herein described; FIG. 24 is a schematic diagram of a technical architecture overview of the Cable Modem Manager according to one embodiment of the systems and methods herein described; FIG. 25 is a schematic diagram of a screen display which may be generated by the Cable Modem Manager to display data according to one embodiment of the systems and methods herein described; FIG. 26 shows a schematic view of where the network management system sits in relation to interfaces for use by users, including customers, service operators and network operators, and the services and network elements being managed; FIG. 27 shows a schematic view of the primary components of the network management system; FIG. 28 shows a more detailed view of components of the network management system together with tools and other systems with which it interacts; FIG. 29 shows a simulator for use in the network management system of FIG. 26 and its connection into a network to be tested; FIG. 30 shows a screen view from a graphical user interface reviewing an alarm by means of the network management system, with access available to a knowledge management system for further analysis; FIG. 31 shows a screen view from a graphical user interface reviewing an alarm in terms of user impact; FIG. 32 shows a screen view from a graphical user interface reviewing an alarm with reference to its network location, with access available to the knowledge management system for further analysis; FIG. 33 shows a screen view from a graphical user interface providing equipment information, with access available to a knowledge management system for further analysis; FIG. 34 shows the options available for a screen view layout as shown in FIG. 33 ; FIG. 35 shows a screen view from a graphical user interface providing an alarm list with access to the knowledge management system for further analysis of selected alarms; FIG. 36 shows a screen view similar to that of FIG. 30 in which the knowledge management system has been accessed for further relevant information; FIG. 37 shows the options available for a screen view layout as shown in FIG. 36 ; FIG. 38 shows a login screen for a user accessing the knowledge management system; FIG. 39 shows a screen view available to a user of the knowledge management system; FIG. 40 shows a further screen view available to a user of the knowledge management system; FIG. 41 shows the options available for a screen view layout as shown in FIG. 39 or 40 ; FIG. 42 shows a network context for the network management system in which a global infrastructure provider controls the end-to-end network; FIG. 43 shows a network context for the network management system in which a service provider uses the network management system to control the service provider's part of the network; FIG. 44 shows a sample Hybrid Fibre-Coax manager deployment according to one embodiment of the systems and methods described herein; FIG. 45 illustrates CMTS router and CMTS card relationships according to one embodiment of the systems and methods described herein; FIG. 46 shows an BFC manager user interface overview for a preferred embodiment of the systems and methods described herein; FIG. 47 illustrates a Knowledge Management System integrated with Imagine Service Emulation Agent modules according to one embodiment of the systems and methods described herein; FIG. 48 illustrates a graphical User Interface structure for a preferred Knowledge Management System according to one embodiment of the systems and methods described herein; FIG. 49 is an XML section of command script for the switch configuration for use with a preferred ISEA according to one embodiment of the systems and methods described herein; FIG. 50 is an XML section of command script for a cable modem configuration for use with a preferred ISEA according to one embodiment of the systems and methods described herein; FIG. 51 is an XML section of command script for a scheduler manager configuration for use with a preferred ISEA according to one embodiment of the systems and methods described herein; FIG. 52 is an XML section of command script for a DHCP protocol configuration for use with a preferred ISEA according to one embodiment of the systems and methods described herein; FIG. 53 is an. XML section of command script for use with a preferred ISEA to configure a scenario named ‘Test’ for one location, containing three services; provisioning, internet access, mail (ISP) according to one embodiment of the systems and methods described herein; FIG. 54 is a continuation of the XML section of command script of FIG. 53 according to one embodiment of the systems and methods described herein; FIG. 55 illustrates scenario organization for an ISEA according to one embodiment of the systems and methods described herein; FIG. 56 shows a preferred hardware architecture for an ISEA according to one embodiment of the systems and methods described herein; FIG. 57 illustrates the six key components comprising a preferred ISEA architecture according to one embodiment of the systems and methods described herein; detailed-description description="Detailed Description" end="lead"? |
Macrolide efflux genetic assembly |
Macrolide resistance associated with macrolide efflux (mef) in Streptococcus pneumoniae has been defined with respect to the genetic structure and dissemination of a novel mefE-containing chromosomal insertion element. The mefEgene is found on the 5′-end of a 5.5 kb or 5.4 kb insertion designated mega (macrolide efflux genetic assembly) found in at least four distinct sites of the pneumococcal genome. The element is transformable and confers macrolide resistance to susceptible S. pneumoniae. The first two open reading frames (ORFs) of the element form an operon composed of mefE and a predicted ATP-binding cassette homologous to msrA. Convergent to this efflux operon are three ORFs with homology to stress response genes of Tn5252. Mega is related to the recently described mefA-containing element Tn1207.1, but lacks genes necessary for transposition and has unique termini and insertion sites. Macrolide resistance due to mega has rapidly increased by clonal expansion of bacteria containing it and horizontally by transformation of previously sensitive bacteria. |
1. An isolated nucleic acid molecule comprising a nucleotide sequence of a macrolide efflux genetic assembly, wherein nucleotide sequence has at least about 70% nucleotide sequence identity with SEQ ID NO:1, and wherein the macrolide efflux genetic assembly determines resistance to macrolide and streptogramin B antibiotics. 2. The isolated nucleic acid molecule of claim 1 wherein the macrolide to which resistance is determined is at least one of erythromycin, clarithromycin and azithromycin. 3. The isolated nucleic acid molecule of claim 1 wherein the streptogramin B to which resistance is determined is at least one of dalfopristin and quinupristin. 4. The isolated nucleic acid molecule of claim 3 wherein the streptogramin B resistance is determined by a nucleotide sequence encoding a protein of the amino acid sequence given in SEQ ID NO:3. 5. The isolated nucleic acid molecule of claim 1 wherein said nucleotide sequence is SEQ ID NO:1. 6. A method of identifying a composition which inhibits efflux of a macrolide and/or a streptogramin B in a bacterium comprising a nucleotide sequence of a macrolide efflux genetic assembly, wherein nucleotide sequence has at least about 70% nucleotide sequence identity with SEQ ID NO:1, and wherein the macrolide efflux genetic assembly determines resistance to macrolide and streptogramin B antibiotics; said method comprising the steps of: (a) contacting a bacterium comprising a nucleotide sequence of a macrolide efflux genetic assembly, wherein nucleotide sequence has at least about 70% nucleotide sequence identity with SEQ ID NO:1, with a test composition and determining whether the test composition inhibits the growth of the bacterium in the absence of a macrolide or a streptogramin B antibiotic; (b) contacting the bacterium with a macrolide or a streptogramin B antibiotic at a subinhibitory concentration; (c) contacting the bacterium with a macrolide or a streptogramin B antibiotic and the test composition and determining the effect on growth of the bacterium, whereby a composition with inhibits efflux of a macrolide or a streptogramin B antibiotic inhibits growth of the bacterium when contacted together with a macrolide or streptogramin B antibiotic at a concentration which is subinhibitory in the absence of the test composition. 7. The method of claim 6 wherein the macrolide to which resistance is determined is at least one of erythromycin, clarithromycin and azithromycin. 8. The method of claim 6 wherein the streptogramin B to which resistance is determined is at least one of dalfopristin and quinupristin. 9. The method of claim 6 wherein said nucleotide sequence is SEQ ID NO:1. 10. The method of claim 6 wherein the bacterium is a gram positive bacterium. 11. The method of claim 10 wherein the bacterium is a species of Streptococcus, Staphylococcus or Enterococcus. 12. A method of predicting resistance to a macrolide antibiotic and/or a streptogramin B antibiotic in a patient infected with a gram positive microorganism, said method comprising the steps of: (a) obtaining a sample comprising nucleic acid from a patient infected with a gram positive microorganism or obtaining a sample comprising nucleic acid from a bacterial sample cultured from a patient infected with a gram positive organism; (b) contacting the sample with a nucleic acid molecule having a sequence of at least 20 contiguous nucleotides derived from the nucleotide sequence of a macrolide efflux genetic assembly, wherein nucleotide sequence has at least about 70% nucleotide sequence identity with SEQ ID NO:1, wherein the nucleic acid molecule has a sequence unique to the macrolide efflux genetic assembly and is not shared with ABC transporter coding seuqences, and wherein the macrolide efflux genetic assembly determines resistance to macrolide and streptogramin B antibiotics; (c) observing binding of the nucleic acid molecule with the sample comprising nucleic acid from the patient or from the bacterial sample cultured from the patient, whereby a prediction of resistance to a macrolide antibiotic and/or streptogramin B antibiotic is made when there is binding to the nucleic acid molecule. 13. The method of claim 12 wherein the macrolide to which resistance is predicted is at least one of erythromycin, clarithromycin and azithromycin. 14. The method of claim 12 wherein the streptogramin B to which resistance is predicted is at least one of dalfopristin and quinupristin. 15. The method of claim 12 wherein said nucleotide sequence of the macrolide efflux genetic assembly is SEQ ID NO:1. 16. The method of claim 12 wherein the gram positive bacterium is a species of Streptococcus, Staphylococcus or Enterococcus. 17. The method of claim 12 wherein the binding observed in step (c) is monitored as a hybridization reaction. 18. The method of claim 12 wherein the binding observed in step (c) results in a product of a polymerase chain reaction. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The field of the present invention includes molecular biology, in particular, with respect to the genetics and molecular mechanism of antibiotic resistance, especially macrolide resistance, in bacteria, and to methods for identifying inhibitors of macrolide resistance, in particular, that resistance to macrolides and/or streptogramins due to efflux systems. Infections caused by Gram positive bacteria including but not limited to Streptococcus pneumoniae remain a devastating and worldwide health problem. The treatment of infections due to S. pneumoniae has become increasingly complicated due to the rapid emergence of resistance to penicillin. While macrolides (including, but not limited to, erythromycin, clarithromycin, azithromycin) are considered alternatives to penicillin for non-meningeal infections, resistance has rapidly emerged to these agents as well. There are two known mechanisms of macrolide resistance in S. pneumoniae , target modification and macrolide efflux. The pneumococcal ermAM gene product, methylates highly conserved adenine residues in the peptidyl transferase center of newly synthesized 23S rRNA [6]. This methylation blocks the binding of macrolides, lincosamides and streptogramins B, thereby conferring the MLS B phenotype of antibiotic resistance [7]. The pneumococcal ermAM gene is associated with conjugative transposons that co-harbor other antimicrobial resistance determinants, e.g., tetM [8]. Until 1996 the only known form of macrolide resistance in pneumococci was due to a methylase (the ermAM gene product) which methylates highly conserved adenine residues in the peptidyl transferase center, domain V, of newly synthesized 23S rRNA. This methylation blocks the binding of macrolides, lincosamides and streptogramin B. The expression of this MLS B phenotype can be inducible or constitutive. There are at least eight classes of erm; ermA, B and C are found on the transposons found in several clinically important pathogens. The pneumococcal ermAM gene is associated with large conjugative and composite transposons, which coharbor other antimicrobial resistance determinants, e.g., tetM and cat. The ermAM gene is very similar to ermB [2, 24]; ermA and C have not been described in S. pneumoniae . Erythromycin MICs for pneumococci containing ermAM are typically =64 μg/ml. In certain other organisms, inactivation is an additional potential mechanism. In 1996, a newly detected macrolide efflux mechanism, mefE, was identified in macrolide-resistant strains of S. pneumoniae lacking an ermAM determinant [9]. Pneumococcal strains containing mefE were reported to express resistance only to 14- and 15-membered macrolides (M phenotype). A 3.7 kb pneumococcal fragment containing mefE when cloned in E. coli was reported to encode a proton motive force-driven transporter sufficient to confer the M phenotype [10]. mefE and the related determinant mefA, originally described in S. pyogenes , are ˜90% identical and have been placed in a single class of macrolide efflux genes [11,12]. Both mefE and mefA are now found in S. pneumoniae [13,14] and are known to be transferable [15]. In Europe, the ermAM determinant is reported to account for recent increases in macrolide resistance of S. pneumoniae while mefA and mefE are found less often [16]. In contrast, most macrolide-resistant pneumococcal strains in North America harbor mefE [13,17-19]. In metropolitan Atlanta, between 1994 and 1999, macrolide resistance of invasive pneumococcal isolates increased from 16.4% to 31.5% [13]. By 1999, mefE was found in 26% of all invasive S. pneumoniae isolated in metropolitan Atlanta [13]. There is a need in the art for an understanding of the mechanisms of antibiotic resistance and the spread thereof, and for methods for identifying inhibitors of resistance mechanisms as well as for identifying compounds which evade the resistance mechanisms to allow for efficacious treatment of infectious diseases of bacterial origin. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides a novel genetic element, i.e., the macrolide efflux genetic assembly (mega), derived originally from erythromycin-resistant Streptococcus pneumoniae . The nucleotide sequence of a specifically exemplified mega element is provided in SEQ ID NO:1, and the amino acid sequences of the proteins encoded by the ORFs therein are given in SEQ ID NO:2-6. SEQ ID NO:2 is the amino acid sequence of a macrolide efflux protein (MefE), and SEQ ID NO:3 is the amino acid sequence of a macrolide and streptogramin B efflux protein (MeI). SEQ ID NOs:4-6 are the amino acid sequences of proteins having significant sequence identity to proteins of conjugative transposons from Gram positive bacteria. The sequences can be used to identify the present of macrolide and or macrolide and streptogramin resistance in a bacterial strain or they can be used in epidemiological studies, for example, to follow the spread of antibiotic resistance. The present invention further provides methods for identifying inhibitors of the MefE and/or MeI efflux proteins. The methods involve determining the growth of a bacterium which produces a MefE and/or a MeI protein. When the bacterium is grown in the presence of a subinhibitory (but non-zero concentration of a macrolide), the bacterium contains a low level of the macrolide in the cytoplasm, without growth inhibition. If there is a compound present in the growth medium which inhibits the efflux protein(s), the concentration within the bacterium rises, and growth is inhibited. Desirably, the macrolide efflux protein is the MefE protein of mega as specifically exemplified herein and the macrolide and streptogramin B efflux protein is the MeI protein of mega as specifically exemplified herein. The mega-containing strain and strains containing derivatives of mega in which either the MefE or the MeI coding sequences have been functionally inactivated (for example, by deleting at least one half of the coding sequence of interest) are tested to determine the subinhibitory concentration of a macrolide and/or a streptogramin. Then test compounds are incorporated in the growth medium in addition to the macrolide and/or streptogramin at varying concentration. A control medium contains the test compounds to verify that those test compounds alone do not inhibit growth of the bacteria. Where the test compound results in inhibition of growth only in the presence of a subinhibitory concentration of a macrolide, there is a conclusion that the test compound causes inhibition of the efflux mechanism. The present invention further provides nucleotide sequences which can be used to predict antibiotic resistance in gram positive infections, for example, those caused by streptococcal, enterococcal or staphylococcal pathogens. Lactococci may also harbor mega and serve as reservoirs of antibiotic resistance which can be transmitted to other Gram positive bacteria, including bacilli, clostridia and various pathogenic microorganisms. The mega-derived mefE nucleotide sequences are characteristic of macrolide resistance (erythromycin, clarithromycin, azithromycin, for example). The presence of mega-derived mel sequences are indicative of resistance to macrolides and streptogramin B antibiotics (including but not limited to dalfopristin and quinupristin). Nucleotides derived in sequence from the 5′ and 3′ termini of mega can be used to determine the presence of the mega element. Bacterial isolates cultured from patient specimens or patient specimens can be used to test for the presence of these characteristic sequences. Nucleic acid molecules, especially single-stranded nucleic acid molecules, which are derived in sequence from SEQ ID NO:1, can be incorporated into PCR assays for related sequences or they can be incorporated into DNA:DNA hybridization assays or RNA:DNA hybridization. Microarray/microchip technology is particularly suited for use in the detection of mega and/or the prediction of antibiotic resistance in a patient infected with a Gram positive microorganism, especially a Gram positive coccus, and as particularly appropriate to such screening and detection, a streptococcal pathogen, e.g., S. pyogenes or S. pneumoniae. As a further aspect of the present invention, the mega element of the present invention can also serve as a “carrier” for the introduction of heterologous DNA into a streptococcal strain, by inserting the DNA of interest into a region of the element not essential for incorporation into the streptococcal genome (or for selecting for its presence). As an alternative to introducing an antibiotic resistance, one can readily delete the mefE and mel sequences and replace them with another selectable marker or replace with a marker for whose expression there is a convenient screen, such as a luciferase coding sequence expressible in the streptococcus or a beta-galactose, beta-glucuronidase, etc. For example, a coding sequence for an antigen of interest operably linked to a promoter expressible in a streptococcal strain can be introduced so that the genetically modified strain can produce the antigen of interest and serve as a vaccine, especially where the streptococcal strain is nonpathogenic or attenuated with respect to virulence while maintaining the ability to colonize a human or animal of interest. |
Isolated human transporter proteins, nucleic acid molecules encoding human transporter proteins, and uses thereof |
The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the transporter peptides of the present invention. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the transporter peptides, and methods of identifying modulators of the transporter peptides. |
1. An isolated peptide consisting of an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids. 2. An isolated peptide comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids. 3. An isolated antibody that selectively binds to a peptide of claim 2. 4. An isolated nucleic acid molecule consisting of a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d). 5. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d). 6. A gene chip comprising a nucleic acid molecule of claim 5. 7. A transgenic non-human animal comprising a nucleic acid molecule of claim 5. 8. A nucleic acid vector comprising a nucleic acid molecule of claim 5. 9. A host cell containing the vector of claim 8. 10. A method for producing any of the peptides of claim 1 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the peptides are expressed from the nucleotide sequence. 11. A method for producing any of the peptides of claim 2 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the peptides are expressed from the nucleotide sequence. 12. A method for detecting the presence of any of the peptides of claim 2 in a sample, said method comprising contacting said sample with a detection agent that specifically allows detection of the presence of the peptide in the sample and then detecting the presence of the peptide. 13. A method for detecting the presence of a nucleic acid molecule of claim 5 in a sample, said method comprising contacting the sample with an oligonucleotide that hybridizes to said nucleic acid molecule under stringent conditions and determining whether the oligonucleotide binds to said nucleic acid molecule in the sample. 14. A method for identifying a modulator of a peptide of claim 2, said method comprising contacting said peptide with an agent and determining if said agent has modulated the function or activity of said peptide. 15. The method of claim 14, wherein said agent is administered to a host cell comprising an expression vector that expresses said peptide. 16. A method for identifying an agent that binds to any of the peptides of claim 2, said method comprising contacting the peptide with an agent and assaying the contacted mixture to determine whether a complex is formed with the agent bound to the peptide. 17. A pharmaceutical composition comprising an agent identified by the method of claim 16 and a pharmaceutically acceptable carrier therefor. 18. A method for treating a disease or condition mediated by a human transporter protein, said method comprising administering to a patient a pharmaceutically effective amount of an agent identified by the method of claim 16. 19. A method for identifying a modulator of the expression of a peptide of claim 2, said method comprising contacting a cell expressing said peptide with an agent, and determining if said agent has modulated the expression of said peptide. 20. An isolated human transporter peptide having an amino acid sequence that shares at least 70% homology with an amino acid sequence shown in SEQ ID NO:2. 21. A peptide according to claim 20 that shares at least 90 percent homology with an amino acid sequence shown in SEQ ID NO:2. 22. An isolated nucleic acid molecule encoding a human transporter peptide, said nucleic acid molecule sharing at least 80 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or 3. 23. A nucleic acid molecule according to claim 22 that shares at least 90 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or 3. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Transporters Transporter proteins regulate many different functions of a cell, including cell proliferation, differentiation, and signaling processes, by regulating the flow of molecules such as ions and macromolecules, into and out of cells. Transporters are found in the plasma membranes of virtually every cell in eukaryotic organisms. Transporters mediate a variety of cellular functions including regulation of membrane potentials and absorption and secretion of molecules and ion across cell membranes. When present in intracellular membranes of the Golgi apparatus and endocytic vesicles, transporters, such as chloride channels, also regulate organelle pH. For a review, see Greger, R. (1988) Annu. Rev. Physiol. 50:111-122. Transporters are generally classified by structure and the type of mode of action. In addition, transporters are sometimes classified by the molecule type that is transported, for example, sugar transporters, chlorine channels, potassium channels, etc. There may be many classes of channels for transporting a single type of molecule (a detailed review of channel types can be found at Alexander, S. P. H. and J. A. Peters: Receptor and transporter nomenclature supplement. Trends Pharmacol. Sci., Elsevier, pp. 65-68 (1997) and http://www-biology.ucsd.edu/˜msaier/transport/titlepape2.html. The following general classification scheme is known in the art and is followed in the present discoveries. Channel-type transporters. Transmembrane channel proteins of this class are ubiquitously found in the membranes of all types of organisms from bacteria to higher eukaryotes. Transport systems of this type catalyze facilitated diffusion (by an energy-independent process) by passage through a transmembrane aqueous pore or channel without evidence for a carrier-mediated mechanism. These channel proteins usually consist largely of a-helical spanners, although b-strands may also be present and may even comprise the channel. However, outer membrane porin-type channel proteins are excluded from this class and are instead included in class 9. Carrier-type transporters. Transport systems are included in this class if they utilize a carrier-mediated process to catalyze uniport (a single species is transported by facilitated diffusion), antiport (two or more species are transported in opposite directions in a tightly coupled process, not coupled to a direct form of energy other than chemiosmotic energy) and/or symport (two or more species are transported together in the same direction in a tightly coupled process, not coupled to a direct form of energy other than chemiosmotic energy). Pyrophosphate bond hydrolysis-driven active transporters. Transport systems are included in this class if they hydrolyze pyrophosphate or the terminal pyrophosphate bond in ATP or another nucleoside triphosphate to drive the active uptake and/or extrusion of a solute or solutes. The transport protein may or may not be transiently phosphorylated, but the substrate is not phosphorylated. PEP-dependent, phosphoryl transfer-driven group translocators. Transport systems of the bacterial phosphoenolpyruvate:sugar phosphotransferase system are included in this class. The product of the reaction, derived from extracellular sugar, is a cytoplasmic sugar-phosphate. Decarboxylation-driven active transporters. Transport systems that drive solute (e.g., ion) uptake or extrusion by decarboxylation of a cytoplasmic substrate are included in this class. Oxidoreduction-driven active transporters. Transport systems that drive transport of a solute (e.g., an ion) energized by the flow of electrons from a reduced substrate to an oxidized substrate are included in this class. Light-driven active transporters. Transport systems that utilize light energy to drive transport of a solute (e.g., an ion) are included in this class. Mechanically-driven active transporters. Transport systems are included in this class if they drive movement of a cell or organelle by allowing the flow of ions (or other solutes) through the membrane down their electrochemical gradients. Outer-membrane porins (of b-structure). These proteins form transmembrane pores or channels that usually allow the energy independent passage of solutes across a membrane. The transmembrane portions of these proteins consist exclusively of b-strands that form a b-barrel. These porin-type proteins are found in the outer membranes of Gram-negative bacteria, mitochondria and eukaryotic plastids. Methyltransferase-driven active transporters. A single characterized protein currently falls into this category, the Na+-transporting methyltetrahydromethanopterin:coenzyme M methyltransferase. Non-ribosome-synthesized channel-forming peptides or peptide-like molecules. These molecules, usually chains of L- and D-amino acids as well as other small molecular building blocks such as lactate, form oligomeric transmembrane ion channels. Voltage may induce channel formation by promoting assembly of the transmembrane channel. These peptides are often made by bacteria and fungi as agents of biological warfare. Non-Proteinaceous Transport Complexes. Ion conducting substances in biological membranes that do not consist of or are not derived from proteins or peptides fall into this category. Functionally characterized transporters for which sequence data are lacking. Transporters of particular physiological significance will be included in this category even though a family assignment cannot be made. Putative transporters in which no family member is an established transporter. Putative transport protein families are grouped under this number and will either be classified elsewhere when the transport function of a member becomes established, or will be eliminated from the TC classification system if the proposed transport function is disproven. These families include a member or members for which a transport function has been suggested, but evidence for such a function is not yet compelling. Auxiliary transport proteins. Proteins that in some way facilitate transport across one or more biological membranes but do not themselves participate directly in transport are included in this class. These proteins always function in conjunction with one or more transport proteins. They may provide a function connected with energy coupling to transport, play a structural role in complex formation or serve a regulatory function. Transporters of unknown classification. Transport protein families of unknown classification are grouped under this number and will be classified elsewhere when the transport process and energy coupling mechanism are characterized. These families include at least one member for which a transport function has been established, but either the mode of transport or the energy coupling mechanism is not known. Ion Channels An important type of transporter is the ion channel. Ion channels regulate many different cell proliferation, differentiation, and signaling processes by regulating the flow of ions into and out of cells. Ion channels are found in the plasma membranes of virtually every cell in eukaryotic organisms. Ion channels mediate a variety of cellular functions including regulation of membrane potentials and absorption and secretion of ion across epithelial membranes. When present in intracellular membranes of the Golgi apparatus and endocytic vesicles, ion channels, such as chloride channels, also regulate organelle pH. For a review, see Greger, R. (1988) Annu. Rev. Physiol. 50:111-122. Ion channels are generally classified by structure and the type of mode of action. For example, extracellular ligand gated channels (ELGs) are comprised of five polypeptide subunits, with each subunit having 4 membrane spanning domains, and are activated by the binding of an extracellular ligand to the channel. In addition, channels are sometimes classified by the ion type that is transported, for example, chlorine channels, potassium channels, etc. There may be many classes of channels for transporting a single type of ion (a detailed review of channel types can be found at Alexander, S. P. H. and J. A. Peters (1997). Receptor and ion channel nomenclature supplement. Trends Pharmacol. Sci., Elsevier, pp. 65-68 and http://www-biology.ucsd.edu/˜msaier/transport/toc.html. There are many types of ion channels based on structure. For example, many ion channels fall within one of the following groups: extracellular ligand-gated channels (ELG), intracellular ligand-gated channels (ILG), inward rectifying channels (INR), intercellular (gap junction) channels, and voltage gated channels (VIC). There are additionally recognized other channel families based on ion-type transported, cellular location and drug sensitivity. Detailed information on each of these, their activity, ligand type, ion type, disease association, drugability, and other information pertinent to the present invention, is well known in the art. Extracellular ligand-gated channels, ELGs, are generally comprised of five polypeptide subunits, Unwin, N. (1993), Cell 72: 31-41; Unwin, N. (1995), Nature 373: 37-43; Hucho, F., et al., (1996) J. Neurochem. 66: 1781-1792; Hucho, F., et al., (1996) Eur. J. Biochem. 239: 539-557; Alexander, S. P. H. and J. A. Peters (1997), Trends Pharmacol. Sci., Elsevier, pp. 4-6; 36-40; 42-44; and Xue, H. (1998) J. Mol. Evol. 47: 323-333. Each subunit has 4 membrane spanning regions: this serves as a means of identifying other members of the ELG family of proteins. ELG bind a ligand and in response modulate the flow of ions. Examples of ELG include most members of the neurotransmitter-receptor family of proteins, e.g., GABAI receptors. Other members of this family of ion channels include glycine receptors, ryandyne receptors, and ligand gated calcium channels. The Voltage-gated Ion Channel (VIC) Superfamily Proteins of the VIC family are ion-selective channel proteins found in a wide range of bacteria, archaea and eukaryotes Hille, B. (1992), Chapter 9: Structure of channel proteins; Chapter 20: Evolution and diversity. In: Ionic Channels of Excitable Membranes, 2nd Ed., Sinaur Assoc. Inc., Pubs., Sunderland, Mass.; Sigworth, F. J. (1993), Quart. Rev. Biophys. 27: 1-40; Salkoff, L. and T. Jegla (1995), Neuron 15: 489-492; Alexander, S. P. H. et al., (1997), Trends Pharmacol. Sci., Elsevier, pp. 76-84; Jan, L. Y. et al., (1997), Annu. Rev. Neurosci. 20: 91-123; Doyle, D. A, et al., (1998) Science 280: 69-77; Terlau, H. and W. Stühmer (1998), Naturwissenschaften 85: 437-444. They are often homo- or heterooligomeric structures with several dissimilar subunits (e.g., a1-a2-d-b Ca 2+ channels, ab 1 b 2 Na + channels or (a) 4 -b K + channels), but the channel and the primary receptor is usually associated with the a (or a1) subunit. Functionally characterized members are specific for K + , Na + or Ca 2+ . The K + channels usually consist of homotetrameric structures with each a-subunit possessing six transmembrane spanners (TMSs). The al and a subunits of the Ca 2+ and Na + channels, respectively, are about four times as large and possess 4 units, each with 6 TMSs separated by a hydrophilic loop, for a total of 24 TMSs. These large channel proteins form heterotetra-unit structures equivalent to the homotetrameric structures of most K + channels. All four units of the Ca 2+ and Na + channels are homologous to the single unit in the homotetrameric K + channels. Ion flux via the eukaryotic channels is generally controlled by the transmembrane electrical potential (hence the designation, voltage-sensitive) although some are controlled by ligand or receptor binding. Several putative K + -selective channel proteins of the VIC family have been identified in prokaryotes. The structure of one of them, the KcsA K + channel of Streptomyces lividans, has been solved to 3.2 Å resolution. The protein possesses four identical subunits, each with two transmembrane helices, arranged in the shape of an inverted teepee or cone. The cone cradles the “selectivity filter” P domain in its outer end. The narrow selectivity filter is only 12 Å long, whereas the remainder of the channel is wider and lined with hydrophobic residues. A large water-filled cavity and helix dipoles stabilize K + in the pore. The selectivity filter has two bound K + ions about 7.5 Å apart from each other. Ion conduction is proposed to result from a balance of electrostatic attractive and repulsive forces. In eukaryotes, each VIC family channel type has several subtypes based on pharmacological and electrophysiological data. Thus, there are five types of Ca 2+ channels (L, N, P, Q and T). There are at least ten types of K + channels, each responding in different ways to different stimuli: voltage-sensitive [Ka, Kv, Kvr, Kvs and Ksr], Ca 2+ -sensitive [BK Ca , IK Ca and SK Ca ] and receptor-coupled [K M and K ACh ]. There are at least six types of Na + channels (I, II, III, μ1, H1 and PN3). Tetrameric channels from both prokaryotic and eukaryotic organisms are known in which each a-subunit possesses 2 TMSs rather than 6, and these two TMSs are homologous to TMSs 5 and 6 of the six TMS unit found in the voltage-sensitive channel proteins. KcsA of S. lividans is an example of such a 2 TMS channel protein. These channels may include the K Na (Na + -activated) and K Vol (cell volume-sensitive) K + channels, as well as distantly related channels such as the Tok1 K + channel of yeast, the TWIK-1 inward rectifier K + channel of the mouse and the TREK-1 K + channel of the mouse. Because of insufficient sequence similarity with proteins of the VIC family, inward rectifier K + IRK channels (ATP-regulated; G-protein-activated) which possess a P domain and two flanking TMSs are placed in a distinct family. However, substantial sequence similarity in the P region suggests that they are homologous. The b, g and d subunits of VIC family members, when present, frequently play regulatory roles in channel activation/deactivation. Animal Inward Rectifier K + Channel (IRK-C) Family IRK channels possess the “minimal channel-forming structure” with only a P domain, characteristic of the channel proteins of the VIC family, and two flanking transmembrane spanners (Shuck, M. E., et al., (1994), J. Biol. Chem. 269: 24261-24270; Ashen, M. D., et al., (1995), Am. J. Physiol. 268: H506-H511; Salkoff, L. and T. Jegla (1995), Neuron 15: 489-492; Aguilar-Bryan, L., et al., (1998), Physiol. Rev. 78: 227-245; Ruknudin, A., et al., (1 998), J. Biol. Chem. 273: 14165-14171). They may exist in the membrane as homo- or heterooligomers. They have a greater tendency to let K + flow into the cell than out. Voltage-dependence may be regulated by external K + , by internal Mg 2+ , by internal ATP and/or by G-proteins. The P domains of IRK channels exhibit limited sequence similarity to those of the VIC family, but this sequence similarity is insufficient to establish homology. Inward rectifiers play a role in setting cellular membrane potentials, and the closing of these channels upon depolarization permits the occurrence of long duration action potentials with a plateau phase. Inward rectifiers lack the intrinsic voltage sensing helices found in VIC family channels. In a few cases, those of Kir1.1a and Kir6.2, for example, direct interaction with a member of the ABC superfamily has been proposed to confer unique functional and regulatory properties to the heteromeric complex, including sensitivity to ATP. The SUR1 sulfonylurea receptor (spQ09428) is the ABC protein that regulates the Kir6.2 channel in response to ATP, and CFTR may regulate Kir1.1a. Mutations in SUR1 are the cause of familial persistent hyperinsulinemic hypoglycemia in infancy (PHHI), an autosomal recessive disorder characterized by unregulated insulin secretion in the pancreas. Eag Potassium Channels The novel human protein, and encoding gene, provided by the present invention is a human ortholog of the rat Eag potassium channel protein, isoform 2 (“Eag2”), which is a member of the ether-a-go-go (Eag) family of voltage-gated potassium channels. The protein sequence of rat Eag2 is 70% identical to that of rat Eag1; another 10% of amino acid differences between Eag2 and Eag1 are conservatively replaced residues (Ludwig et al., Mol Cell Neurosci 2000 July;16(1):59-70). Eag2 voltage-gated potassium channels are significantly activated at voltages near −100 mV. Eag2 expresses outward-rectifying, non-inactivating voltage-dependent potassium currents similar to those of Eag1. However, Eag2 currents begin to activate at subthreshold potentials that are 40-50 mV more negative than the subthreshold potentials required for Eag1 activation. Thus, Eag2 channels activate at strongly negative voltages and do not inactivate, thereby providing continuous outward currents even at the most negative membrane potentials known to exist in neurons (Saganich et al., J Neurosci 1999 Dec. 15;19(24):10789-802). Potassium channels that are open at very negative membrane potentials, such as Eag2 channels, control the subthreshold activities of neurons. For example, such channels are a factor in the resting potential and play a role in regulating the degree of excitability of neurons by modulating the affects of synaptic inputs and the threshold for action potential initiation. Furthermore, such channels can play key roles in modulating cell behavior even when the channels are present at low concentrations because few conductances are active at such voltages (Saganich et al., J Neurosci 1999 Dec. 15; 19(24):10789-802). Rat eag2 mRNA is primarily expressed in neural tissue. Furthermore, expression of Eag2 mRNA in the brain is extremely unique in that expression is concentrated in layer IV neurons of the cerebral cortex, which are the main recipients of the thalamocortical input. This unique cortical expression pattern suggests that Eag2 channels play important roles in cortical processing. Specifically, Eag2 channels are likely to be important for modulating behavioral state-dependent entry of sensory information to the cerebral cortex (Saganich et al., J Neurosci 1999 Dec. 15;19(24):10789-802). Transporter proteins, particularly members of the Eag potassium channel subfamily, are a major target for drug action and development. Accordingly, it is valuable to the field of pharmaceutical development to identify and characterize previously unknown transport proteins. The present invention advances the state of the art by providing previously unidentified human transport proteins. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is based in part on the identification of amino acid sequences of human transporter peptides and proteins that are related to the Eag potassium channel subfamily, as well as allelic variants and other mammalian orthologs thereof. These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate transporter activity in cells and tissues that express the transporter. Experimental data as provided in FIG. 1 indicates expression in humans in the brains of infants. |
Packaging machine for cigarettes |
For the production of (cigarette) packs of different configurations, for example standard pack (13), round-edged pack (14) or octagonal pack (15), subassemblies and elements of the packaging machine are exchanged and/or uncoupled from the drive. For this purpose, a pack-specific operating element is provided with a drive which has a coupling and/or uncouplable gear-mechanism parts. These, in turn, are assigned operable handling elements which allow adjustment for coupling or uncoupling or removing sub-elements. |
1. A packaging machine, in particular for producing cigarette packs, having folding subassemblies and elements for folding or shaping blanks (28) and having conveying elements for transporting packaging material and (partly) finished packs, characterized in that in the case of production changeover, in particular in respect of size and/or configuration of the (cigarette) pack, it is possible for folding subassemblies and/or elements and/or conveying elements to be wholly or partially exchanged or uncoupled from the drive. 2. The packaging machine as claimed in claim 1, characterized in that the correct exchange of folding subassemblies and/or shaping subassemblies and/or conveying elements can be checked by sensors, in particular by contactless initiators (23, 24), which respond to associated contact components on the elements which have been, or can be, exchanged. 3. The packaging machine as claimed in claim 2, characterized in that the sensors, in particular two initiators (23, 24) assigned to each adjustable subassembly which can be uncoupled or exchanged, are connected to a central control means (27), and in that production operation is only allowed when all the elements or subassemblies have been correctly exchanged and/or removed from the drive. 4. The packaging machine as claimed in claim 1, characterized in that a folding turret (17) can be removed as a whole from the drive and replaced by another folding turret (17), in particular a plate-like folding turret (17) which can be rotated about a vertical axis and is mounted in a releasable manner at a top end of an upright shaft (18), there being fitted on the folding turret, preferably on the underside, contact protrusions—contact rings (25, 26)—with which contact can be made by (two) initiators (23, 24) positioned beneath the same. 5. The packaging machine as claimed in claim 1, characterized in that processing elements, in particular shaping tools for producing or preparing round edges, have a gear mechanism (32, 56) for executing operating movements of the tools, and in that the tools, including the respectively associated gear mechanism (32, 56), can be uncoupled from the drive in order to stop operation of the tools. 6. The packaging machine as claimed in claim 5, characterized in that the gear mechanism (32, 56) for executing the movements of the elements, tools or the like is assigned a further gear mechanism, namely an intermediate gear mechanism (34) or a preliminary gear mechanism (58), which is connected in each case to the drive, and in that the drive can be uncoupled from the drive in the region of the intermediate gear mechanism (34) or of the preliminary gear mechanism (58). 7. The packaging machine as claimed in claim 1, characterized in that a drive shaft or actuating shaft (40) is connected to a driven element, in particular to a pivoting lever (41), via a coupling (48, 49) which can be actuated from the outside, and in that the driven element—pivoting lever (41)—can be uncoupled from the drive by virtue of the coupling being released. 8. The packaging machine as claimed in claim 7, characterized in that the driven element, in particular the pivoting lever (41), is mounted on the actuating shaft by way of an adjustable mount, in particular a sleeve (43), and in that the coupling (48, 49) can be actuated by axial displacement of the sleeve (43). 9. The packaging machine as claimed in claim 8, characterized in that the sleeve (43) or the like can be displaced in the axial direction by an adjusting wheel (44) which can be operated from the outside, and in that one coupling part (48) is connected to the sleeve and a corresponding coupling part (49) is connected to the actuating shaft (40). 10. The packaging machine as claimed in claim 5, characterized in that, in order to uncouple a subassembly or element from the drive, a drive wheel, in particular an intermediate wheel (60) designed as a gearwheel, can be disengaged from adjoining gearwheels, in particular disengaged from the gearwheel (59) assigned to the gear mechanism (56), by axial displacement. 11. The packaging machine as claimed in claim 10, characterized in that the gearwheel or intermediate wheel (60) can be adjusted in the axial direction by a tool which can be actuated from the outside, namely an actuating element (64). 12. The packaging machine as claimed in claim 1, characterized in that it is possible to remove elements or tools, in particular scoring rollers (72, 73) for stamping blanks (28) or material webs (71), preferably by virtue of a shaft or spindle (69, 70) which bears the scoring rollers (72, 73) being removed. 13. The packaging machine as claimed in claim 12, characterized in that the spindles (69, 70) which bear the elements, in particular scoring rollers (72, 73), have an uncouplable spindle component (77, 78), the spindle components (77, 78) being mounted, preferably by way of conical coupling ends (81, 82), in corresponding recesses of rotatable carrying components (79, 80), which are mounted in a stationary manner. 14. The packaging machine as claimed in claim 1, characterized in that the elements which can be exchanged or brought to a standstill have elevations (50), thickened portions (85) or the like adjacent to one or more initiators (23, 24), it being possible for one initiator (23) to be activated in one position and for the other initiator (24) to be activated in another position. 15. The packaging machine as claimed in claim 1, characterized in that, once uncoupled from the drive, rotatable elements can be fixed in a predetermined relative position, in particular by a fixing pin (68) entering into bores. |
Datachable plug coupling provided with an additional locking element |
The detachable plug coupling serves to connect liquid lines, particularly fuel lines of motor vehicles. The plug coupling is provides a tubular insertion part (3) with an encircling remaining rib (4) and a coupling housing (1) with a cylindrical accommodating space (2) for the insertion part (3). The plug coupling also has an enlarged head part (6) with a separate retaining element (7), which is provided with elastically opening retaining edges (10). The retaining edges are radially directed into the accommodating space (2) and engage behind the retaining rib (4) after the insertion part (3) has been pushed in. In order to ensure that the retaining rib (4) is correctly snapped in behind the retaining edges (10) after the insertion part (3) has been inserted, an additional locking element (17) is provided, which is guided in a transversally displaceable manner on the housing head (6) between an opening position and a closing position. |
1-5. (canceled) 6. A locking element for a detachable plug coupling for the connection of liquid lines, said detachable plug coupling having a tubular insertion part (3) with a circumferential retaining rib (4); a coupling housing (1) with an accommodating space (2); an enlarged housing head (6) having a separate retaining element (7) that includes elastically opening retaining edges (10) oriented radially into the accommodating space (2), said retaining edges operative to engage behind the retaining rib (4) after the insertion of the insertion part (3) into the accommodating space (2), said locking element comprising: two lateral walls (18) that contact on both sides of the housing head (6); a U-shaped front plate (20) connecting the two lateral walls; a transverse wall (19) on 4s a bottom side of the locking element; and inward pointing edges on a side opposite the front p)late (20), the inward pointing edges operative to be engaged on a head edge (23) of the coupling housing (1) such that the head edge (23) is guided between two end positions in a manner that allows for transverse shifting wherein the front plate (20), when in an open position, presents a first semicircular recess (25) corresponding to the external diameter of the retaining rib (4) and a second semicircular recess (26) according to the tube diameter of the insertion part (3), the second semicircular recess extends from a base of the first semicircular recess (25) in the direction to the transverse wall (19), and wherein on the front plate (20), at each edge of the first semicircular recess (25), before the transition to the second semicircular recess (26), an arc-shaped support bar (27) is formed, extending into the accommodating space (2), where support surfaces (28) of the support bars, in the loaded state, end with an engagement plane of the retaining edges (10), said locking element (17) adapted to be connected to the housing head (6) and operative to be moved transversely to the insertion direction in front of the retaining rib (4) after the correct engagement of the insert part (3) in the retaining element (7). 7. The locking element of claim 6, wherein on the internal side of the front plate (20), slanted position humps (29) are formed on both sides such that in the open position of the locking element (17), the slanted position humps are located in front of an external edge (31) of the housing head (6), and, during the transverse shifting into a closed position, the slanted position humps elastically open to slide over the front side of the housing head (6), and again engage on the external edge (31) of the housing opening. 8. The locking element of claim 6 wherein the retaining edges (10) of the retaining element (7) are formed on supporting bodies (11) that are insertable into corresponding recesses (12) in the housing wall (16) when the retaining element is installed and wherein ends of the support bodies (11) are connected by V-shaped spring bridges (14) connected with pressure plates (15) that project outward through corresponding openings (13) in the housing wall (16), said pressure plates operative to be pressed in from the outside to detach the coupling such that the two lateral walls (18) of the locking element (17) present recesses (32), said recesses (32) operative to release the pressure plates (15) when the locking element is in the open position and cover the pressure plates at least partially when the locking element is in the closed position. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Such a plug coupling with an additional locking element is known, for example, from WO 96/34224. The locking element is here designed as a catch which is attached in a manner so it can be pivoted via a hinge which is formed on the external surface of the coupling housing. A locking tongue is formed on the catch which, during engagement by pivoting of the catch after the engagement of the insertion part, is arranged between a slit trap which reaches into the housing, and the retaining rib in order to prevent the insertion part from being pulled out of the accommodating space of the coupling housing. This locking element, in its constructive design, is unsatisfactory because it requires, for its attachment and to achieve the desired locking effect to the coupling housing, various constructive changes and adaptations. In addition, the manufacture of the hinge connection also requires an additional work step that increases the manufacturing cost of the entire plug coupling. Finally, it is considered a drawback that the catch prior to the pivoting projects transversely upward and thus occupies a large amount of space in the open position. The object of the invention is to design the above described locking element for use with the existing plug coupling housing in such a manner that the modification of the plug coupling to add the locking element is possible with minimal constructive effort and without any changes to the coupling housing and so that the handling properties of the additionally applied locking element in the delivered state are not negatively affected. This objective is substantially solved by the present invention which can be pressed without problem onto the head of the coupling housing in the axial direction and easily shifted into the locking position after the establishment of the coupling connection with the insertion part. In this process, it is ensured by means of the positioning humps on the internal side of the front wall that the locking element is secured both in its opening position and in its locking position. Additionally, the locking element can be used for retrofitting existing coupling housings without a great effort, even if the coupling housings are already in use. |
<SOH> SUMMARY OF THE INVENTION <EOH>The plug coupling of the present invention includes of a tubular insertion part having a circumferential retaining rib which can be connected to the end of one fuel line and a coupling housing is provided for connecting to the end of the other fuel line. The housing is formed with a cylindrical accommodating space and an enlarged housing head with a separate retention element. The retention element is provided with elastically opening retaining edges which are oriented radially into the accommodating space. The retaining edges engage behind the retaining rib after the insertion of the insertion part into the accommodating space. The plug coupling also includes a locking element which can be connected to the head of the coupling housing. The locking element is operative to be moved transversely to the insertion direction in front of the retaining rib after the correct engagement of the insertion part in the retention element. |
Communications operating system and method therefor |
A mobile subscriber unit is provided with a memory (8) containing the cell selection and re-selection functions relating to a plurality of different radio networks (4 to 6) eg. GSM, UMTS, satelite, available in its locality, thus enabling the subscriber unit to identify and camp on to any one of the networks. Once a communications link is established, the subscriber unit requests over-the-air download of the full operating protocol from the chosen network. A self-check procedure is optionally provided to ensure that the downloaded data is fuctioning correctly. The choice of network may be influenced by user preference data held in a memory (10). The invention enables operation of a subscriber unit on a choice of many networks without the need for a large memory for holding the full protocols for all available networks. |
1. A user equipment for enabling a user to access one of a plurality of radio communications networks (3-6), the user equipment including; a detector (1) for detecting at least one of the plurality of radio communications networks (3-6), a first store (8) for storing network identification data and operational variables relevant to each of said plurality of networks, a second store (10) for storing data relevant to user preferences, an identifier (7) for identifying detected networks by comparison of received signal characteristics with identification data stored in the first store (8), a first selector (9) for choosing one of the detected and identified networks by comparison with the data stored in the second store (10), a second selector (11) for selecting operational variables stored in the first store (8) and pertaining to the chosen network, means (12) for setting up a communications channel with the chosen network using the selected operational variables, and means for receiving (2) an enabling program, thereby to enable communications with the chosen network. 2. A user equipment according to claim 1 and further including means (12) for requesting the enabling program from the chosen network. 3. A user equipment according to claim 2 and further including a third store (13) for storing the enabling program. 4. A user equipment according to claim 1 and further including a module (13) containing the enabling program. 5. A user equipment according to claim 1 in which the first store (8) includes cell selection and re-selection functions for each of the plurality of radio communications networks (3-6). 6. A user equipment according to claim 1 and further including means (8) for performing a self-check procedure. 7. A method of enabling a user equipment to access one of a plurality of radio communications networks, the method including the steps of; detecting at least one of said networks, identifying (14) the detected networks, choosing (15,16) one of the detected and identified networks by comparison with stored user data, selecting operational variables pertaining to the chosen network from a network variable settings store, setting up (18) a communications channel with the chosen network using the selected operational variables, and receiving (21) an enabling program, thereby enabling communication between the user equipment and the chosen network. 8. A method according to claim 7 and including the further steps of requesting (20) the enabling program from the chosen network and storing (21) said enabling program within said user equipment. 9. A method according to claim 7 in which the stored user data relates to network performance capabilities. 10. A method according to claim 7 and including the further step of performing a self check procedure (22). |
<SOH> DISCUSSION OF THE BACKGROUND <EOH>Within the past several years, radio frequency (RF) communication systems have progressed from a very limited number of systems, each available for one type of function, to a vast number of potential systems available for providing the same type of basic service. For example, cellular communication systems began in Europe with the Nordic mobile telephone (NMT) system. At the time, this was the only system which provided mobile dial-up telephone operations. Since its introduction, newer analogue systems have been introduced, such as total access communication system (TACS), and more recently, digital systems such as group special mobile (GSM) and the third generation system (3G) often referred to as UMTS (Universal Mobile Telecommunications System) and currently under standardisation. All these systems have several features which distinguish themselves from each other. For example, GSM and UMTS not only operate on different carrier frequencies but also use different multiple access systems. GSM uses a time division multiple access system and UMTS uses a code division multiple access system. Conversely, there are features common to both systems, with both GSM and UMTS being cellular systems whereby a cell is supported by a single base station (in GSM terminology), or node B (in UMTS terminology). Several base stations or node B's are normally controlled by a base station controller or radio network controller, respectively. The multiple access technologies referred to above permit multiple subscribers to each of these systems to access a serving base station simultaneously by means of a portable subscriber unit often referred to as a mobile station or a user equipment. Until recently, one problem for the end user was that to be able to go anywhere in the world and use a subscriber unit, the user needed to carry different types of subscriber unit, since one unit alone would not operate on all of the systems. However, multi-mode and re-configurable user equipments are now becoming commercially available, allowing the user to select any one of several available communication systems, eg GSM or UMTS. Ideally, a re-configurable user equipment should work on a wide range of systems and standards and particularly, should be able to detect and identity alternative communications networks available in its locality. GB-A-2294844 discloses a method for enabling a user equipment to access one of many different networks, by initially accessing a common radio system which is in communication with each of the different networks. The common radio system provides to the user equipment, a list of networks available. The user equipment chooses one of the networks on the list and requests an enabling program from the common radio system so that communications between the user equipment and the chosen network may be set up directly. A limitation of this known method is that it does not allow direct access of one of a plurality of networks by a user equipment, ie. it requires the provision of a common radio system. |
<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the present invention, there is provided a user equipment for enabling a user to access one of a plurality of radio communications networks, the user equipment including; a detector for detecting at least one of the plurality of radio communications networks, a first store for storing network identification data and operational variables relevant to each of said plurality of networks, a second store for storing data relevant to user preferences, an identifier for identifying detected networks by comparision of received signal characteristics with identification data stored in the first store, a first selector for choosing one of the detected and identified networks by comparison with the data stored in the second store, a second selector, for selecting operational variables stored in the first store and pertaining to the chosen network, means for setting up a communications channel with the chosen network using the selected operational variables, and means for receiving an enabling program thereby to enable communications with the chosen network. According to a second aspect of the present invention there is provided a method of enabling a user equipment to access one of a plurality of radio communications networks, the method including the steps of; detecting at least one of said networks, identifying the detected networks, choosing one of the detected and identified networks by comparison with stored user data, selecting of operational variables pertaining to the chosen network from a network variable settings store, setting up a communications channel with the chosen network using the selected operational variables, and receiving an enabling program thereby enabling communications between the user equipment and the chosen network. The enabling program may be requested from the chosen network by the user equipment and subsequently received over the communications channel and downloaded and stored into a dedicated store in the user equipment. Alternatively, the enabling program may be downloaded into the user equipment by means of an appropriate plug-in card, or module, which is connected to the user equipment by the user once the network selection has been made and notified to the user (by means of a visual display, for example). The network identification data may relate to characteristics of the signals received by the detector from the various networks, and may comprise, for example, carrier frequency, multiple access type, pilot signal frequency. The user preferences in the second store may be set, for example, so that a detected UMTS network is always selected in preference to a GSM network. Alternatively, the stored preferences may include a preferred received signal strength, available data rate or quality of service, in which case the first selector chooses a detected and identified network which meets the preferred performance criteria. In this latter example, the identifier is adapted to determine such criteria from the detected signals from each network. The user preferences may be entered into the user equipment by the user with the aid of a key pad. Alternatively, a stored default list of preferences could be used or the user could select options from a pre-stored list. The operational variables stored in the first store are sufficient to enable the user equipment to set up the communications channel with the selected network and preferably comprise the so-called “cell selection and re-selection functions” of each of the plurality of networks. Once the channel has been established, the enabling program, ie., the full protocol for registering with and enabling communications with a chosen network, may be downloaded over the communications channel and stored in the user equipment. Optionally, the user equipment may also perform a self-check for checking that the enabling program has been downloaded successfully and that the user equipment has been re-configured correctly as a result of the download. Advantageously, the invention enables the user equipment to identify and set up a communications link with an available network directly, without going through any common system, and also enables the user equipment to monitor other available networks whilst registered with the chosen network. The invention has the further advantage in that no extra pilot channel is required by each network for imparting network data to a user equipment. Further, the user equipment is not required to permanently store the full enabling program for every available network. Such a requirement would necessitate very large amounts of memory. The enabling program which is downloaded is a reduced protocol rather than a full protocol, therefore, download time is minimised and spectral efficiency is maximised. |
Preparation for the prophylaxis of restenosis |
The invention relates to a preparation for restenosis prevention. The preparations for restenosis prevention known as yet do not reach sufficient active agent concentrations in the affected sections of the vascular walls as higher doses cause undesirable side effects. The present invention is a preparation to which at least one antihyperplastic agent is added that has a distribution ratio between butanol and water ≧0.5. The lipophilic active agent is absorbed by the vascular wall fast and in sufficient quantity. The preparation may be a liquid that can pass through capillaries and may contain a contrast agent so that the active agent is transferred into the vascular wall without any additional effort while the usually required contrast radiograms are taken. The preparation may also be applied to a catheter. |
1. A composition of matter comprising a pharmaceutically effective amount of taxol or a tumor-active analog thereof, solubilized in a pharmaceutically acceptable carrier comprising an oil having a dipole moment of between about 0.5 Debyes and about 2.0 Debyes. 2. The composition of claim 1 wherein said oil comprises an ether lipid as a major component. 3. The composition of claim 1 wherein said oil is a marine oil having an ether lipid as a major component thereof. 4. The composition of claim 1 wherein said oil comprises a terpene hydrocarbon as a major component. 5. The composition of claim 1 which comprises taxol. 6. An emulsion comprising the composition of claim 1. 7. The emulsion of claim 6 that is an oil-in-water emulsion. 8. A composition of matter comprising a pharmaceutically effective amount of taxol solubilized in an oil selected from the group consisting of orange roughy oil, shark liver oil, squalane and squalene. 9. The composition of claim 8 wherein said oil is squalane or squalene. 10. A composition of matter comprising an oil having a dipole moment of between about 0.5 Debyes and about 2.0 Debyes in which taxol or a tumor-active analog thereof is solubilized in said oil at a level greater than or equal to 1 mg/ml. 11. The composition of matter of claim 10 wherein said taxol or tumor-active analog thereof is solubilized in said oil at a level greater than or equal to 5 mg/ml. 12. The composition of matter of claim 11 wherein said oil is squalane or squalene. 13. A method for solubilizing taxol or a tumor-active taxol analog in a pharmaceutically acceptable carrier comprising: (a) solubilizing taxol or said taxol analog in an anhydrous alcohol; (b) mixing the solution of (a) with a sufficient amount of a pharmaceutically acceptable carrier comprising an oil having a dipole moment of between about 0.5 Debyes and about 2.0 Debyes, to solubilize said solution. 14. The method of claim 13 further comprising removing said anhydrous alcohol from the solution of step (b). 15. A pharmaceutical composition comprising the solution prepared by the method of claim 14. 16. The method of claim 14 further comprising preparing an oil-in-water emulsion with the solution from which said anhydrous alcohol has been removed. 17. An emulsion prepared by the method of claim 16. 18. A pharmaceutical dosage form comprising the composition of claim 1. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Taxol is a poorly water soluble alkaloid isolated from several species of Western Yew. Taxol exhibits antimitotic properties and is presently undergoing phase I clinical trials for the treatment of cancers. Taxol has been shown to be active against leukemia, colon, breast, melanoma, sarcomas, and Lewis lung tumor systems. Tarr et al. (1987) Pharm. Res. 4:162-165; Horwitz (1992) TIPS 13:134-136. In vitro studies indicate that concentrations of taxol of 0.1-10.0 μ/ml stabilize microtubules, thus disrupting normal cell division. Rowinsky et al. (1990) J. Natl. Cancer Inst. 82:1247-1259. Taxol is a complex diterpene having a taxane ring system with a four-membered oxetane ring and an ester sidechain at position C-13, as follows: In an attempt to increase taxol's solubility and develop more feasible clinical formulations, investigators have acylated carbons at the 7-position and 10-position of the taxene ring. These efforts have yielded compounds that retain anti-tumor activity. Rowinsky et al. (1990) J. Natl. Cancer Inst. 82:1247-1259. Because of its poor solubility in water and many oils, taxol has been administered in formulations using Cremophors. Cremophors are polyoxyethylated castor oils. The current, most widely used Sigma taxol formulation consists of ethanol:Cremophor EL™:isotonic saline (5:5:90). The drug's solubility in this vehicle does not exceed 0.6 mg/ml and it remains physically stable only for a short time (3 hr). Therefore, in view of the limited solubility, large volumes of these formulations must be infused to obtain a desired total dose of 30 mg. Tarr et al. (1987) Pharm. Res. 4:162-165. A patient is usually required to check into a hospital and endure intravenous infusion for an extended period, such as twenty-four hours. Typically, taxol is administered intravenously in a preparation containing 30 μg/ml over a period of twenty-four hours, followed by a week of rest and another dose. This course of administration is typically repeated two more times. Further, the BASF Cremophor EL™ (polyoxyethylated castor oil) is extremely toxic and in dogs has been shown to produce vasodilation, labored breathing, lethargy, hypotension and death. Rowinsky et al. (1990) J. Natl. Cancer Inst. 82:1247-1259. Anaphylactoid reactions observed in phase I trials of the taxol formulation in Cremophor EL™ were attributed to the rapid administration of the Cremophor. Grem et al. (1987) Cancer Treat. Rep. 71:1179-1184. Hypersensitivity reactions have also been observed using the above formulation; one patient had a fatal reaction. It is unclear whether taxol itself or the Cremophor vehicle is principally responsible for these hypersensitivity reactions. Rowinsky et al. (1990) J. Natl. Cancer Inst. 82:1247-1259. Water-miscible cosolvents have also been used in taxol preparations but require infusion times even longer than the currently used formulation. Drugs formulated with cosolvents may precipitate if infused too fast. Yalkowsky et al. (1977) Drug Intell. Clin. Pharm. 11:417-419. To avoid using the toxic Cremophor and provide a non-precipitating I.V. taxol formulation, Tarr et al. (1987) Pharm. Res. 4:162-165 formulated taxol with Intralipid (trademark of RabiVitrum [formerly Cutter Medical]), a commonly used parenteral emulsion comprising soybean oil, lecithin, egg yolk phospholipids, and glycerol. This vehicle was found unsuitable for pharmaceutic use because of the low solubility of taxol in soybean oil (0.3 mg/ml). Tarr et al. then made taxol emulsion formulations using triacetin, a water-soluble triester. Taxol is soluble in triacetin to a level of about 75 mg/ml. A 50% triacetin emulsion containing 20 mg/ml taxol as well as the emulsifying agents L-alpha-lecithin, pluronic F-86 (BASF), polysorbate 80 (Sigma Chemical Co.), and ethyl oleate was tested as a possible bolus I.V. formulation. Glycerol was added to the triacetin emulsion to slow creaming, but was found to have little effect on emulsion stability. Administration of the 50% triacetin formulation caused toxic reactions including lethargy, ataxia and respiratory depression in animal models, presumably due to the toxicity of the triacetin. The 50% triacetin emulsion showed an intravenous LD50 of 1.2 ml/kg in mice. The triacetin emulsion initially had a 1 μm average diameter particle size which slowly increased and finally exhibited instability, separating into two phases at six months. Vigorous shaking again formed an emulsion having an average droplet size of 2 μm. Triacetin is 1,2,3-triacetylglycerol: As taxol has been determined to be an especially effective anti-cancer agent, formulations which do not contain toxic ingredients and which allow delivery of pharmaceutically relevant dosages in a reasonable period of time, such as orally or by injection, are especially needed. Such formulations have not been previously available. Methods and compositions for solubilizing pharmaceutically relevant dosages of taxol in pharmaceutically acceptable oils are therefore highly desirable objects of this invention. The use of oil-in-water emulsions for delivery of taxol and its tumor-active analogs are needed to avoid problems of precipitation of I.V. solutions at the time of administration to increase bioavailability of orally administered taxol and to prevent gastrointestinal upset. In non-emulsified form, taxol is degraded in the stomach. However, prior efforts to produce pharmaceutically acceptable emulsions containing taxol have failed due to both the relative insolubility of taxol in typical pharmaceutically suitable oils and the requirements for toxic surface-active agents. Pharmaceutically acceptable oils useful in forming oil-in-water emulsions are well-known to the art and include mineral, vegetable, animal and marine oils. However, the low solubility of taxol in many oils, such as safflower, olive and soybean oils (about 0.3-0.6 mg/ml) which have fatty acid glycerides and triglycerides as major components, has prevented the use of such oils in taxol formulations. Marine oils, especially those that comprise ether lipids rather than ester lipids and terpene hydrocarbons, are known to the art and include, among many others, orange roughy, squalane, squalene and shark liver oil. |
<SOH> SUMMARY OF THE INVENTION <EOH>This invention provides compositions of matter comprising a pharmaceutically effective amount of taxol or a tumor-active analog thereof solubilized in a pharmaceutically acceptable carrier comprising an oil having a dipole moment of between about 0.5 Debyes and about 2.0 Debyes, and preferably between about 1.6 and 1.7 Debyes. Oils from deep-water marine organisms are preferred. Oils comprising ether lipids as major components are preferred. Ether lipids can be glycerol ethers, including unsaturated glycerol ethers and polyunsaturated glycerol ethers. Oils comprising terpene based hydrocarbons derived from marine oils, like squalene and squalane, are also preferred for preparation of taxol solutions. The compositions of the present invention show a many-fold increase (up to five hundred times) of oral absorption over the prior art formulation using the EL Cremophor™ (BASF). This invention provides high concentration solutions of taxol or tumor-active taxol analogs in oils having greater than or equal to about 1 mg/ml of taxol dissolved therein. These taxol solutions are useful in the preparation of pharmaceutical emulsions and other dosage forms. Marine oils are preferred taxol solvents, particularly those oils comprising ether lipids including unsaturated and polyunsaturated glycerol ethers. High concentration taxol solutions can also be prepared with oils comprising terpene-based hydrocarbons, like squalane and squalene. Non-toxic derivatives of squalene and squalane that have dipole moments between about 0.5 Debyes and 2.0 Debyes are also useful in preparation of high concentration taxol solutions and pharmaceutical dosage forms for taxol. Solutions of taxol or tumor-active taxol analogs in the pharmaceutically acceptable oils of this invention may be prepared by directly dissolving taxol in the oil or by forming a first solution by dissolving taxol in a preliminary solvent such as an anhydrous alcohol followed by mixing with the oil and evaporation of the solvent. The solutions of taxol or tumor-active analogs of taxol made by the methods of this invention are preferably used to prepare oil-in-water emulsions for pharmaceutical use in anti-tumor therapy. Emulsions are preferred vehicles for taxol for both intravenous and oral administration and can be prepared by any means known to the art using known surfactants and emulsion stabilizing components from the solutions of taxol in oils of this invention. Oil-in-water emulsions containing taxol can also be made using self-emulsifying glasses prepared as disclosed in U.S. Ser. No. 07/830,058 and subsequent related applications. Such glass compositions comprise a water-soluble, nonsurface active matrix compound and an oil containing solubilized taxol from which emulsions can be readily formed by contacting the glass with an aqueous phase. Melt-spinning technology as described in U.S. Pat. Nos. 5,011,532, 5,034,421 and can also be used to prepare colloidal dispersions for pharmaceutical administration of taxol. Emulsions produced from self-emulsifying glasses or colloidal dispersions produced by melt-spinning technology do not require art-recognized surfactants or emulsifying agents. A pharmaceutically effective emulsion containing taxol is produced from a self-emulsifying glass by mixing it with sufficient aqueous phase to form an emulsion. No emulsive mixing or surfactants are necessary. Emulsions and colloidal dispersions containing therapeutically effective amounts of taxol are administered to patients suffering from cancers against which taxol is known to have a therapeutic effect in appropriate oral or intravenous dosages to effect reduction in the disease. detailed-description description="Detailed Description" end="lead"? |
Method and data processing ssytem for timing the duration of a session |
A method of timing the duration of a session during a connection between a user station and a chargeable zone of an Internet site, during which session the user station can access chargeable data and/or services of the site without disconnecting from the Internet, the method comprising: loading a monitoring program into the user station for execution throughout the duration of the session, said monitoring program being configures, while it is being executed, to send information relating to the state of the connection to a time-metering server, said information being renewed periodically throughout the session. |
1-102. cancelled. 103. A method of timing the duration of a session during a connection between a user station and a chargeable zone of an Internet site, during which session the user station can access at least one of chargeable data and chargeable services of the site without disconnecting from the Internet, the method comprising: loading a monitoring program into the user station for execution throughout the duration of the session, said monitoring program being configured, while it is being executed, to send information relating to the state of the connection to a time-metering server, said information being renewed periodically throughout the session. 104. A method according to claim 103, wherein the information relating to the state of the connection is transmitted to the time-metering server via a TCP socket connection. 105. A method according to claim 103, wherein the monitoring program is contained in a hidden frame. 106. A method according to claim 103, wherein the information relating to the state of the connection is renewed in a period that is shorter than or equal to one minute. 107. A method according to claim 103, wherein further comprising determining a duration for the session, as a function at least of information received by the time-metering server due to execution of the monitoring program. 108. A method according to claim 103, wherein the monitoring program is of the Java applet type. 109. A method according to claim 103, wherein the monitoring program executes as soon as it is loaded into the user station. 110. A method according to claim 103, wherein at least one of the user station and the user is identified prior to the monitoring program being loaded into the user station. 111. A method according to claim 103, wherein throughout a session, the user station periodically receives varying codes of temporary validity as generated by the time-metering server. 112. A method according to claim 111, wherein code validity has a duration of not more than three minutes. 113. A method according to claim 111, wherein the codes are generated by the time-metering server at substantially constant time intervals. 114. A method according to claim 111, wherein the number of codes generated during a session is counted, and wherein the duration of the session is determined as a function at least of the number of codes counted. 115. A method according to claim 111, wherein the site periodically receives varying countersign codes of temporary validity associated with the corresponding varying codes sent to the user station, these countersign codes being generated by the time-metering server, and wherein user station access to the at least one of chargeable data and chargeable services is authorized so long as the code and the corresponding countersign code satisfy a predetermined relationship. 116. A method according to claim 115, wherein the predetermined relationship is an equality relationship. 117. A method according to claim 103, wherein a connection duration counter is incremented each time the time-metering server receives information from the monitoring program. 118. A method according to claim 103, wherein the site includes different pages associated with display temporary addresses. 119. A method according to claim 103, wherein the monitoring program is configured to warn the time-metering server in the event of disconnection from the chargeable zone. 120. A method according to claim 103, wherein the time-metering server is arranged to cease timing the session in the event of no information being received from the monitoring program over a predetermined length of time. 121. A method according to claim 103, wherein the site is configured so as to associate at least one charging rate with at least one of chargeable services and chargeable data. 122. A method according to claim 121, wherein the site includes at least one of chargeable services and chargeable data associated with charging rates stored in a database. 123. A method according to claim 122, wherein the database is stored with the site supplier or with the time-metering server. 124. A method according to claim 121, wherein the site includes pages organized in directories associated with respective charging rates. 125. A method according to claim 121, wherein the site includes pages of addresses that are organized as a function of charging rates, and wherein access to the pages is performed via a virtual host. 126. A method according to claim 103, wherein the site includes a proxy type program serving to replace an URL address request from the user station with another URL address. 127. A method according to claim 126, wherein the proxy type program is configured so as to compare the code with the corresponding countersign code and verify that the predetermined relationship is satisfied. 128. A method according to claim 126, wherein the proxy type program is configured to read information contained in a page sent to the user station and to transmit said information to the time-metering server. 129. A method according to claim 128, wherein the user station sends information to the time-metering server relating to reception of the page, and wherein the time-metering server is configured to compare said information with the information transmitted by the site because of execution of the proxy type program. 130. A method according to claim 129, wherein the time-metering server is configured to start timing a session on receiving the information relating to reception of the page by the user station. 131. A method according to claim 129, wherein the page is arranged to transmit information to the monitoring program and wherein the monitoring program is configured to transmit information relating to reception of the page to the time-metering server. 132. A method according to claim 103, wherein the site includes pages containing information representative of a charging rate, in particular information that is in tag form. 133. A method according to claim 129, wherein the information sent by the proxy type program to the time-metering server comprises a link, wherein the page loaded into the user station sends to the time-metering program a request concerning the link, and wherein this request constitutes information relative to reception of the page by the user station and at least one of starts timing a session and indicates a charging rate change. 134. A method according to claim 103, wherein on each occasion a new page is received by the user station, in particular a new HTML page corresponding to at least one of chargeable data and chargeable service, information relating to said page is sent to the time-metering server. 135. A method according to claim 103, wherein a link to access to a page is replaced by a program in javascript type language, this program being configured to execute as soon as the user activates the link and enable the access to the page, this program being also configured so as to send to the monitoring program information relative to the page loading and possibly to the page charging rate. 136. A method according to claim 135, wherein the monitoring program is configured so as to send said information to the time-metering server. 137. A method according to claim 103, wherein content categories are associated with the charging rates in order to enable to exercise control as a function of age or of parental authorization. 138. A method according to claim 103, wherein at least one of the user station and the user is identified, at least by recognizing a login and a password. 139. A method according to claim 138, wherein the login and the password are identical to the login and the password needed to connect to an Internet access supplier. 140. A method according to claim 103, wherein at least one of the user station and the user is identified by recognizing at least one intrinsic characteristic of the user station. 141. A method according to claim 103, in which the user station has an IP address, and wherein, prior to authorizing access to the chargeable zone, said fP address is compared with one or more prerecorded addresses for identifying the user. 142. A method according to claim 103, wherein the user is identified by information coming from Internet access suppliers. 143. A method according to claim 103, wherein the user station is a mobile terminal with a connection linked to an operator of one of the types GPRS, UMTS, G3, CDPD, this terminal comprising an identifying chip including a code PIN or WIM, and wherein, prior to authorizing access to the chargeable zone, said code PIN or WIM is compared with one or more prerecorded codes for identifying the user. 144. A method according to claim 103, wherein the time-metering server is configured for a given user account to aggregate connection durations to distinct sites, said durations possibly being associated with charging rates. 145. A method according to claim 103, wherein the duration of the connection to the chargeable zone is displayed on the user station. 146. A method according to claim 103, wherein the time-metering server is configured to compare the amount corresponding to the cost of various connections to the chargeable zones as accumulated with an upper limit on expenditure entered by the user, and to warn the user in the event the cost exceeds the upper limit on expenditure. 147. A method according to claim 103, wherein the monitoring program is configured to send information to the time-metering server relating to a data rate at which the user station receives chargeable data. 148. A method according to claim 103, wherein the connection durations of a plurality of users to at least one of chargeable services and chargeable data of a site are recorded, and wherein said durations are subjected to statistical processing. 149. A method according to claim 103, wherein the user station is a personal computer with an Internet connection, in particular of the xDSL type. 150. A method according to claim 103, wherein the user station is suitable for communicating over a wireless link with the Internet network. 151. A method according to claim 103, wherein the time-metering server is arranged to issue a bill on a user account associated with a mobile telephone or with a fixed telephone line or with an electricity meter, or with a bank account or with an account associated with a credit card. 152. A method according to claim 103, wherein the time-metering server is configured in such a manner as to enable a user account to be debited for transactions other than for connection time to a chargeable zone of a site. 153. A method according to claim 103, wherein the site includes a chargeable service for allocating bandwidth capacity. 154. A method according to claim 103, wherein the connection to the chargeable zone is initiated by the user station opening an e-mail sent to the user station and including a link to the chargeable zone of the site. 155. A method of enabling a data medium to be used on an appliance, the data medium being configured in such a manner as to prevent at least part of it from being used independently of the appliance being in connection with a chargeable zone of an Internet site, comprising: enabling the appliance to be connected to the chargeable zone of the Internet site; metering the duration of the connection; and enabling the medium to be used throughout the time the appliance is connected to the chargeable zone. 156. A method according to claim 155, wherein the appliance is selected from the following group: a personal computer, a games console, a video projector, a reader of at least one of audio and video disks. 157. A computer program referred to as a monitoring program for use during connection of a user station to a chargeable zone of an Internet site in order to time the duration of a session during which the user station can access to least one of chargeable data and chargeable services of the site without disconnection from the Internet, the program being configured so that once loaded into said user station it executes during a chargeable session, said monitoring program being configured so that on execution it sends information relating to the state of the connection to a time-metering server, said information being renewed periodically during the session. 158. A program according to claim 157, said program being configured to renew the information relating to the state of the connection within a period of time that is less than or equal to one minute. 159. A program according to claim 157, said program being a Java type applet. 160. A program according to claim 157, said program being configured to send information to the time-metering server relating to a data rate at which chargeable data is received by the user station. 161. A program according to claim 157, said program being arranged to receive varying codes of temporary validity issued by a time-metering server. 162. A program according to claim 157, said program being arranged to warn the time-metering server in the event of disconnection from the chargeable zone. 163. A program according to claim 157, said program being configured to identify the user station by recognizing a PIN or WIM code, the user station being a mobile terminal with a connection linked to an operator of one of the types GPRS, UMTS, G3 pr CDPD and comprising and identifying chip including a PIN or WIM code. 164. A signal transmitted over the Internet network, the signal including data readable by a computer and corresponding to the program as defined in claim 55 being downloaded to a user station. 165. A data processing system including: means for accessing a data processing network which users and at least one of chargeable content and chargeable service providers can log onto, and memory means for storing account details of each user, which data processing system enables a user listed by the system and who has been identified to access chargeable data or services of at least one of a content and a service provider without having to specify means of payment directly to said at least one of content and service provider and determines the duration of access by the user to the at least of one content and service provider and possibly the quantity of chargeable data consulted or transferred and possibly the services consumed by the user and debits the account of the user by a corresponding amount, possibly depending on the nature of said data or services. 166. A system according to claim 165, wherein said data processing network is the Internet and is preferably accessed via a high bit rate connection, in particular an x-DSL or satellite connection. 167. A system according to claim 165, said system applying different charge bands according to at least one of: the identity of the at least one of content and service providers that a user can access via said system, and the nature of the contents or services consulted or consumed. 168. A system according to claim 165, wherein it can be accessed by means of an Internet address, possibly without previously downloading a plug-in. 169. A system according to claim 165, wherein the data processing system is adapted to route a user logging onto the data processing system to a particular content or service provider as a function of predetermined criteria. |
Methods of treating alzheimer's disease |
Disclosed are methods for treating Alzheimer's disease, and other diseases, and/or inhibiting beta-secretase enzyme, and/or inhibiting deposition of A beta peptide in a mammal, by use of 3,4-disubstituted piperidinyl compounds of formula (I) wherein the variables R1, R2, R3, R4, Q, W, X, Z, m, and n are defined herein. |
1. A method of treating or preventing Alzheimer's disease in a patient in need of such treatment comprising administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C)C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6CO—O, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 2. A method of treating Alzheimer's disease in a patient in need of such treatment comprising administering to the patient a compound disclosed in claim 1, or a pharmaceutically acceptable salt thereof. 3. A method of treating Alzheimer's disease by modulating the activity of beta amyloid converting enzyme, comprising administering to a patient in need of such treatment a compound disclosed in claim 1, or a pharmaceutically acceptable salt thereof. 4. The method according to claim 1, further comprising the administration of a P-gp inhibitor, or a pharmaceutically acceptable salt thereof. 5. A method of treating a patient who has, or in preventing a patient from getting, a disease or condition selected from the group consisting of Alzheimer's disease, for helping prevent or delay the onset of Alzheimer's disease, for treating patients with mild cognitive impairment (MCI) and preventing or delaying the onset of Alzheimer's disease in those who would progress from MCI to AD, for treating Down's syndrome, for treating humans who have Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch-Type, for treating cerebral amyloid angiopathy and preventing its potential consequences, i.e. single and recurrent lobar hemorrhages, for treating other degenerative dementias, including dementias of mixed vascular and degenerative origin, dementia associated with Parkinson's disease, dementia associated with progressive supranuclear palsy, dementia associated with cortical basal degeneration, or diffuse Lewy body type of Alzheimer's disease and who is in need of such treatment which includes administration of a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 6. The method according to claim 5 wherein the compound of formula (I) is selected from the group consisting of: 4-[2-[7-[(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethyl]-morpholine; (R)-3-[7-[(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-propane-1,2-diol; (S)-3-[7-[(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-propane-1,2-diol; (R)-3-[2-[7-[(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethoxy]-propane-1,2-diol; (S)-3-[2-[7-[(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethoxy]-propane-1,2-diol; 1-[2-[7-[(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethyl]-4-methyl-piperazine; 1-[(3R,4S-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yl-2-naphthalen-2-yl-ethanone; (3R,4S,5 S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-piperidin-5-ol; 3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-[7-[(R)-2,3-dihydroxy-propoxymethyl]-naphthalen-2-ylmethoxy]-piperidine; (3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-[7-[(S)-2,3-dihydroxy-propoxymethyl]-naphthalen-2-ylmethoxy]-piperidine; (3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-[6-[(R)-2,3-dihydroxy-propoxymethyl]-naphthalen-2-ylmethoxy]-piperidine; (3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-[6-[(S)-2,3-dihydroxy-propoxymethyl]-naphthalen-2-ylmethoxy]-piperidine; 4-[(3R,4S,5 S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-butan-1-ol; 3-[(3R,4S,5 S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-propan-1-ol; 1-{2-[(3R,4R,5 S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-ethyl}-4-methyl-piperazine; (3R,4R,5S)-[4-[4(3-benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-ylmethoxy]-ethyl]-morpholine; (3R,4S,5 S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-(4-methoxy-benzyloxy)-piperidin-5-ol; (3R,4s,5S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3,5-bis-(4-methoxy-benzyloxy)-piperidine; (3S,4R,5R)-4-[2-[4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethoxy]-ethyl]-morpholine; (3S,4R,5R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-methoxymethyl-5-(naphthalen-2-ylmethoxy)-piperidine; (3S,4R,5R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl[3-(4-methyl-piperazin-1-yl)-propyl]-carbamate; (3S,4R,5R)-4-[4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethylsulphanyl]-pyridine; 2-(4-cyclohexyl-butoxy)-5-[(3R,4R)-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-piperidin-4-yl]-pyrimidine; (3′R,4′R)-6-(3-cyclohexyl-propoxy)-3′-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridine; (3S,4R,5R)-[4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-yl]-methanol; (3S,4R,5R)-N-[4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl]-N,N′,N′-trimethyl-ethane-1,2-diamine; (3S,4R,5R)-[4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl]-diethyl-amine; 1-[(3R,4S,5S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(2-morpholin-4-yl-ethoxymethyl)-piperidin-3-yl]-2-naphthalen-2-yl-ethanone; (3R,4R)-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-4-[4-[3-(2-methoxy-phenoxy)-propoxy]-phenyl]-piperidine; (3R,4s,5S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3,5-bis-(3,4,5-trimethoxy-benzyloxy)-piperidine; (3R,4R,5S)-4-[4-(3-benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-5-[1,2,4]triazol-1-ylmethyl-piperidine; (3R,4R)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-3-(quinolin-7-ylmethoxy)-piperidine; 2-(7-{(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl}-naphthalen-2-ylmethoxy)-ethanol; 7-{(3R,4R)-4-[4-(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl}-naphthalen-2-ylmethyl)-dimethyl-amine; (3R,4R)-3-(4-benzyloxy-naphthalen-2-ylmethoxy)-4-(4-fluoro-phenyl)-piperidine; (3′R,4′R)-3′-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-6-[3-(2-methoxybenzyloxy)-propoxy]-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridine; (3R,4R)-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-piperidine; (3S,4R,5R)-1-[4-[4-(3-benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl]-imidazolidin-2-one; (3R,4R)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-3-(2-oxo-1,2-dihydro-quinolin-7-ylmethoxy)-piperidine; (3R,4R)-3-(isoquinolin-7-ylmethoxy)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-piperidine; (3R,4R)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-3-(1,2,3,4-tetrahydro-quinolin-7-ylmethoxy)-piperidine; 1-[2-[7-[(3R,4R)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethyl]-4-methyl-piperazine; 1-[2-[7-[(3R,4S,5 S)-5-hydroxy-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-piperdin-3-yloxymethyl]-naphthalen-2-yloxy]-ethyl]-4-methyl-piperazine; (3R,4S,5S)-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-piperidin-5-ol; (3R,4R,5S)-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-4-{4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl}-5-(1H-tetrazol-5-ylmethyl)-piperidine; (3′S,4′S)-3′-(1,4-dimethoxy-naphthalen-2-ylmethyoxy)-4-[S-(2-methoxy-benzyloxy)-propoxy]-1′,2′,3′,4′,5′,6′-hexahydro-[1,4′]bipyridin-2-one; (3RS,4RS)-2-[4(3-Naphthalen-2-ylmethoxy-piperidin-4-yl)-phenoxy]-ethylthiophene-2-carboxylate hydrochloride; (3RS,4RS)-2-[4(3-Naphthalen-2-ylmethoxy-piperidin-4-yl)-phenoxy]-ethyl 2-chloro-benzoate hydrochloride; (3RS,4RS)-2-[4[3[4(2-methoxy-benzyloxy)-naphthalen-2-ylmethoxy]-piperidin-4-yl]-phenoxy]-ethyl benzoate hydrochloride; (3RS,4RS)-4-[4(3-Benzyloxy-propoxy)-phenyl]3-(2,3-dihydro-benzo[1,4]dioxin-6-ylmethoxy)-piperidine; (3RS,4RS)-3-(Naphthalen-2-ylmethoxy)-4-[4(3-phenyl-[1,2,4]oxadiazol-5-ylmethoxy)-phenyl]-piperidine trifluoroacetate; (3RS,4RS)-3-(Naphthalen-2-ylmethoxy)-4-[4(3-phenyl-isoxazol-5-ylmethoxy)-phenyl]-piperidine trifluoroacetate; (3RS,4RS)-3-(Naphthalen-2-ylmethoxy)-4-[4(3-phenylsulphanyl-propyl)-phenyl]-piperidine; (3RS,4RS)-3-[4[4[2(Benzothiazol-2-ylsulphanyl)-ethyl]-phenyl]-piperidin-3-yloxymethyl]-naphthalen-1-ol; (3RS,4RS,5 SR)-3-(4-Benzyloxy-naphthalen-2-ylmethoxy)-4-(4-fluoro-phenyl)-5-propyl-piperidine; (3SR,4RS,5RS)-4-(4-Benzyloxymethyl-phenyl)-3-methoxymethyl-5-(naphthalen-2-ylmethoxy)-piperidine; (SR)- or (RS)-1-[(3RS,4SR)4-(4-fluoro-phenyl)-piperidin-3-yl]-2-naphthalen-2-yl-ethyl benzoate hydrochloride; (1RS,2RS,3RS,5 SR)-2-(4-Benzyloxy-naphthalen-2-ylmethoxy)-3-(4-fluoro-phenyl)-8-aza-bicyclo[3.2.1]octane; (3RS,4RS)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperdine; 4-[2-[7-[(3RS,4RS)-4-[4(3-Benzyloxy-propoxy)-phenyl]piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethyl]-morpholine hydrochloride (1:2); 3-[7-[(3RS,4RS)-4-[4(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-propane-1,2-diol; (RS)- and (SR)-3-[2-[7-[(3RS,4RS)-4-[4(3-benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethoxy]-propane-1,2-diol hydrochloride (1:1); 1-[2-[7-[(3RS,4RS)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl]-naphthalen-2-yloxy]-ethyl-4-methyl-piperazine hydrochloride (1:3); 1-[(3RS,4SR)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-piperidin-3-yl]-2-naphthalen-2-yl-ethanone hydrochloride (1:1); (3RS,4SR,5SR)-4-[4(3-Benzyloxy-propoxy)-phenyl]-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-piperidin-5-ol; 4-[4-(3-benzyloxy-propoxy)-phenyl]-3-[7-[(RS)2,3-dihydroxy-propoxymethyl]naphthalen-2-ylmethoxy]-piperidine; 4-[4-(3-benzyloxy-propoxy)-phenyl]-3-[6-[(RS)-2,3-dihydroxy-propoxymethyl]-naphthalen-2-ylmethoxy]-piperidine; 4-[(3RS,4SR,5SR)-4-[4(3-Benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-butan-1-ol; 3-[(3RS,4SR,5SR)-4-[4(3-Benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-propan-1-ol; 1-{2-[(3RS,4RS,5SR)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-ethyl}-4-methyl-piperazine; 4-{2-[(3RS,4RS,5SR)-4-[4(3-Benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-piperidin-5-yloxy]-ethyl}-morpholine; (3RS,4SR,5SR)-4-[4(3-Benzyloxy-propoxy)-pheny]-l3-(4-methoxy-benzyloxy)-piperidin-5-ol; (3R,4s,5S)-4-[4(3-Benzyloxy-propoxy)-phenyl]-3,5-bis-(4-methoxy-benzyloxy)-piperidine; (3SR,4RS,5RS)-4-[2-[4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethoxy]-ethyl]-morpholine; (3SR,4RS,5RS)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-3-methoxymethyl-5-(naphthalen-2-ylmethoxy)-piperidine; (3SR,4RS,5RS)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl[3 (4-methyl-piperazin-1-yl)-propyl]-carbamate; (3SR,4RS,5RS)-4-[4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethylsulphanyl]-pyridine; 2-(4-Cyclohexyl-butoxy)-5-[(3RS,4RS)-3-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-piperidin-4-yl]-pyrimidine; (3′RS,4′RS)-6-(3-Cyclohexyl-propoxy)-3′-(1,4-dimethoxy-naphthalen-2-ylmethoxy)-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]-bipyridine; (3SR,4RS,5RS)-[4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-yl]-methanol hydrochloride; (3SR,4RS,5RS)-N-[4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl]-N,N′,N′-trimethyl-ethane-1,2-diamine; (3SR,4RS,5RS)-[4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-yl]-methyldiethyl-amine; 1-[(3RS,4SR,5SR)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(2-morpholin-4-yl-ethoxymethyl)-piperidin-3-yl]-2-naphthalen-2-yl-ethanone; (3RS,4RS)-3-(1,4-Dimethoxy-naphthalen-2-ylmethoxy)-4-[4-[3-(2-methoxy-phenoxy)-propoxy]-phenyl]-piperidine; (3R,4s,5S)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-3,5-bis-(3,4,5-trimethoxy-benzyloxy)-piperidine; (3RS,4RS,5SR)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-3-(naphthalen-2-ylmethoxy)-5-[1,2,4]triazol-1-ylmethyl-piperidine hydrochloride; (3RS,4RS)-4-[4-[3-(2-Methoxy-benzyloxy)-propoxy]-phenyl]-3-(quinolin-7-ylmethoxy)-piperidine; 2-(7-{(3RS,4RS)-4-[4(3-Benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymethyl}-naphthalen-2-ylmethoxy)-ethanol; 7-{(3RS,4RS)-4-[4-(3-Benzyloxy-propoxy)-phenyl]-piperidin-3-yloxymthyl}-naphthalen-2-ylmethyl)-dimethyl-amine; (3R,4R)-3-(4-Benzyloxy-naphthalen-2-ylmethoxy)-4-(4-fluoro-phenyl)-piperidine; (3S,4S)-3-(4-Benzyloxy-naphthalen-2-ylmethoxy)-4-(4-fluoro-phenyl)-piperidine; (3′RS,4′RS)-3′-(1,4-Dimethoxy-naphthalen-2-ylmethoxy)-6-[3-(2-methoxybenzyloxy)-propoxy]-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridine; (3RS,4RS)-3-(1,4-Dimethoxy-naphthalen-2-ylmethoxy)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-piperidine; (3SR,4RS,5RS)-1-[4-[4-(3-Benzyloxy-propoxy)-phenyl]-5-(naphthalen-2-ylmethoxy)-piperidin-3-ylmethyl]-imidazolidin-2-one; (3RS,4RS)-4-[4-[3-(2-Methoxy-benzyloxy)-propoxy]-phenyl]-3-(2-oxo-1,2-dihydro-quinolin-7-ylmethoxy)-piperidine; (3RS,4RS)-3-(Isoquinolin-7-ylmethoxy)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-piperidine; (3RS,4RS)-4-[4-[3-(2-Methoxy-benzyloxy)-propoxy]-phenyl]-3-(1,2,3,4-tetrahydro-quinolin-7-ylmethoxy)-piperidine; and (3RS,4SR,5 SR)-3-(1,4-Dimethoxy-naphthalen-2-ylmethoxy)-4-[4-[3-(2-methoxy-benzyloxy)-propoxy]-phenyl]-piperidin-5-ol. 7. A method of treating or preventing Alzheimer's disease in a patient in need of such treatment comprising administering a therapeutically effective amount of a composition comprising one or more pharmaceutically acceptable carriers and a compound of Formula (I) or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) —Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 8. (canceled) 9. A method for inhibiting beta-secretase activity, comprising contacting an effective amount for inhibition of a compound of formula (I), or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11) —, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11) —, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11) —, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 10. A method for inhibiting cleavage of an amyloid precursor protein (APP) isotype at a site in the APP isotype that is susceptible to cleavage, comprising contacting said APP isotype with an effective cleavage inhibitory amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C)C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6, where R6 is as defined above, (J) —NR6SO2, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11) —, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11) —, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11) —, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 11. A method for inhibiting production of amyloid beta peptide (A beta) in a cell, comprising administering to said cell an effective inhibitory amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 12. The method of claim 11, wherein the cell is an animal cell. 13. The method of claim 12, wherein the animal cell is a mammalian cell. 14. The method of claim 13, wherein the mammalian cell is human. 15. A composition comprising beta-secretase complexed with a compound of formula (I), or a pharmaceutically acceptable salt thereof: where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 16. A method for producing a beta-secretase complex comprising the composition of claim 15. 17. A method for inhibiting the production of beta-amyloid plaque in an animal, comprising administering to said animal an effective inhibiting amount of a compound of formula (I): where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a hetero atom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 18. The method of claim 17, wherein said animal is a human. 19. A method for treating or preventing a disease characterized by beta-amyloid deposits on or in the brain, comprising administering to a patient in need of such treatment or prevention an effective therapeutic amount of a compound of formula (I): where R1 is (I) aryl, or (II) heterocycle; where R2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L1-T1-L2-T2-L3-T3-L4-T4-L5-U, where L1, L2, L3, L4 and L5 are independently chosen from: (A) a bond, (B) C1-8-alkylene, (C) C2-8-alkenylene, (D) C2-8-alkynylene, or (E) are absent, where T1, T2, T3, and T4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR6)—, where R6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR5R6)—, where R5 is as defined for R6, and where R6 is as defined above, (E) —CO—, (F) —CR7R8—, where R7 and R8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO2 groups, (G) —O— or —NR6—, where R6 is as defined above, (H) —S(O)0-2—, (I) —SO2NR6—, where R6 is as defined above, (J) —NR6SO2—, where R6 is as defined above, (K) —CONR6—, where R6 is as defined above, (L) —NR6CO—, where R6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR6—, where R6 is as defined above, (Q) —NR6—CO—NR6—, where R6 are the same or different and are as defined above, (R) —NR6—CO—O—, where R6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R3 and R4 together are a bond, or R4a-Z1-X1— where R4a is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R5R6)—N—(CH2)1-3—, where R5 and R6 are as defined above, (N) (R5R6)—N—, where R5 and R6 are as defined above, (O) lower-alkyl-S(O)0-2—, (P) aryl-S(O)0-2—, (Q) heterocyclyl-S(O)0-2—, (R) HO—SO3— or a salt thereof, (S) H2N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R11)—, where R11 is hydrogen or lower-alkyl, (F) —S(O)0-2—, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R11)O, where R11 is as defined above, (K) —N(R11)—CO—O—, where R11 is as defined above, (L) —CO—N(R11)—, where R11 is as defined above, (M) —N(R11)—CO—, where R11 is as defined above, (N) —N(R11)—CO—N(R11)—, where R11 are the same or different and are as defined above, (O) —CH(OR9)—, where R9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X1 is: (A) a bond, (B) —O—, (C) —N—(R11)—, where R11 is as defined above, (D) —S(O)0-2—, (E) —(CH2)1-3—, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R11—, where R11 is as defined above, (V) —CHOR9—, where R9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR10—, where R10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R1; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R11— and either R2 contains a substituent L1-T1-L2-T2-L3-T3-L4-T4-L5-U or R4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R11— when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond. 20. (canceled) 21. (canceled) |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention is the use of known compounds to treat Alzheimer's disease and other similar diseases, and more specifically to compounds that inhibit beta-secretase, an enzyme that cleaves amyloid precursor protein to produce A beta peptide, a major component of the amyloid plaques found in the brains of Alzheimer's sufferers. 2. Background of the Invention Alzheimer's disease (AD) is a progressive degenerative disease of the brain primarily associated with aging. Clinical presentation of AD is characterized by loss of memory, cognition, reasoning, judgment, and orientation. As the disease progresses, motor, sensory, and linguistic abilities are also affected until there is global impairment of multiple cognitive functions. These cognitive losses occur gradually, but typically lead to severe impairment and eventual death in the range of four to twelve years. Alzheimer's disease is characterized by two major pathologic observations in the brain: neurofibrillary tangles and beta amyloid (or neuritic) plaques, comprised predominantly of an aggregate of a peptide fragment know as A beta. Individuals with AD exhibit characteristic beta-amyloid deposits in the brain (beta amyloid plaques) and in cerebral blood vessels (beta amyloid angiopathy) as well as neurofibrillary tangles. Neurofibrillary tangles occur not only in Alzheimer's disease but also in other dementia-inducing disorders. On autopsy, large numbers of these lesions are generally found in areas of the human brain important for memory and cognition. Smaller numbers of these lesions in a more restricted anatomical distribution are found in the brains of most aged humans who do not have clinical AD. Amyloidogenic plaques and vascular amyloid angiopathy also characterize the brains of individuals with Trisomy 21 (Down's Syndrome), Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch-Type (HCHWA-D), and other neurodegenerative disorders. Beta-amyloid is a defining feature of AD, now believed to be a causative precursor or factor in the development of disease. Deposition of A beta in areas of the brain responsible for cognitive activities is a major factor in the development of AD. Beta-amyloid plaques are predominantly composed of amyloid beta peptide (A beta, also sometimes designated betaA4). A beta peptide is derived by proteolysis of the amyloid precursor protein (APP) and is comprised of 39-42 amino acids. Several proteases called secretases are involved in the processing of APP. Cleavage of APP at the N-terminus of the A beta peptide by beta-secretase and at the C-terminus by one or more gamma-secretases constitutes the beta-amyloidogenic pathway, i.e. the pathway by which A beta is formed. Cleavage of APP by alpha-secretase produces alpha-sAPP, a secreted form of APP that does not result in beta-amyloid plaque formation. This alternate pathway precludes the formation of A beta peptide. A description of the proteolytic processing fragments of APP is found, for example, in U.S. Pat. Nos. 5,441,870; 5,721,130; and 5,942,400. An aspartyl protease has been identified as the enzyme responsible for processing of APP at the beta-secretase cleavage site. The beta-secretase enzyme has been disclosed using varied nomenclature, including BACE, Asp, and Memapsin. See, for example, Sindha et al., 1999 , Nature 402:537-554 (p501) and published PCT application WO00/17369. Several lines of evidence indicate that progressive cerebral deposition of beta-amyloid peptide (A beta) plays a seminal role in the pathogenesis of AD and can precede cognitive symptoms by years or decades. See, for example, Selkoe, 1991 , Neuron 6:487. Release of A beta from neuronal cells grown in culture and the presence of A beta in cerebrospinal fluid (CSF) of both normal individuals and AD patients has been demonstrated. See, for example, Seubert et al., 1992 , Nature 359:325-327. It has been proposed that A beta peptide accumulates as a result of APP processing by beta-secretase, thus inhibition of this enzyme's activity is desirable for the treatment of AD. In vivo processing of APP at the beta-secretase cleavage site is thought to be a rate-limiting step in A beta production, and is thus a therapeutic target for the treatment of AD. See for example, Sabbagh, M., et al., 1997 , Alz. Dis. Rev. 3, 1-19. BACE1 knockout mice fail to produce A beta, and present a normal phenotype. When crossed with transgenic mice that over express APP, the progeny show reduced amounts of A beta in brain extracts as compared with control animals (Luo et al., 2001 Nature Neuroscience 4:231-232). This evidence further supports the proposal that inhibition of beta-secretase activity and reduction of A beta in the brain provides a therapeutic method for the treatment of AD and other beta amyloid disorders. At present there are no effective treatments for halting, preventing, or reversing the progression of Alzheimer's disease. Therefore, there is an urgent need for pharmaceutical agents capable of slowing the progression of Alzheimer's disease and/or preventing it in the first place. Compounds that are effective inhibitors of beta-secretase, that inhibit beta-secretase-mediated cleavage of APP, that are effective inhibitors of A beta production, and/or are effective to reduce amyloid beta deposits or plaques, are needed for the treatment and prevention of disease characterized by amyloid beta deposits or plaques, such as AD. |
<SOH> SUMMARY OF INVENTION <EOH>The present invention relates to a method of treating a patient who has, or in preventing a patient from developing, a disease or condition selected from the group consisting of Alzheimer's disease, for helping prevent or delay the onset of Alzheimer's disease, for helping to slow the progression of Alzheimer's disease, for treating patients with mild cognitive impairment (MCI) and preventing or delaying the onset of Alzheimer's disease in those who would progress from MCI to AD, for treating Down's syndrome, for treating humans who have Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch-Type, for treating cerebral amyloid angiopathy and preventing its potential consequences, i.e. single and recurrent lobar hemorrhages, for treating other degenerative dementias, including dementias of mixed vascular and degenerative origin, dementia associated with Parkinson's disease, dementia associated with progressive supranuclear palsy, dementia associated with cortical basal degeneration, or diffuse Lewy body type of Alzheimer's disease and who is in need of such treatment which comprises administration of a therapeutically effective amount of a compound of formula (I): where R 1 is (I) aryl, or (II) heterocycle; where R 2 is (I) phenyl, (II) naphthyl, (III) acenaphthyl, (IV) cyclohexyl, (V) pyridyl, (VI) pyrimidinyl, (VII) pyrazinyl, (VIII) oxo-pyridinyl, (IX) diazinyl, (X) triazolyl, (XI) thienyl, (XII) oxazolyl, (XIII) oxadiazolyl, (XIV) thiazolyl, (XV) pyrrolyl, or (XVI) furyl, optionally substituted by one, two, or three of halogen, hydroxy, cyano, trifluoromethyl, lower-alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, lower-alkoxy-lower-alkyl, cyano-lower-alkyl, carboxy-lower-alkyl, lower-alkanoyloxy-lower-alkyl, lower-alkoxycarbonyloxy-lower-alkyl, lower-alkoxycarbonyl, lower alkoxy groups, lower-alkylenedioxy, or L 1 -T 1 -L 2 -T 2 -L 3 -T 3 -L 4 -T 4 -L 5 -U, where L 1 , L 2 , L 3 , L 4 and L 5 are independently chosen from: (A) a bond, (B) C 1-8 -alkylene, (C) C 2-8 -alkenylene, (D) C 2-8 -alkynylene, or (E) are absent, where T 1 , T 2 , T 3 , and T 4 are independently chosen from: (A) a bond, (B) —CH(OH)—, (C) —CH(OR 6 )—, where R 6 is hydrogen, lower-alkyl, lower-alkenyl, aryl-lower-alkyl, acyl, or together with the heteroatom to which they are attached are a 5- or 6-membered heterocyclic ring which can contain an additional N, O, or S atom or a SO or SO 2 group and the additional N atom if present can be optionally substituted by lower-alkyl, (D) —CH(NR 5 R 6 )—, where R 5 is as defined for R 6 , and where R 6 is as defined above, (E) —CO—, (F) —CR 7 R 8 —, where R 7 and R 8 together with the C atom to which they are attached form a three, four, five, six, or seven membered ring which can contain one or two O or S atoms or SO or SO 2 groups, (G) —O— or —NR 6 —, where R 6 is as defined above, (H) —S(O) 0-2 —, (I) —SO 2 NR 6 —, where R 6 is as defined above, (J) —NR 6 SO 2 —, where R 6 is as defined above, (K) —CONR 6 —, where R 6 is as defined above, (L) —NR 6 CO—, where R 6 is as defined above, (M) —O—CO—, (N) —CO—O—, (O) —O—CO—O—, (P) —O—CO—NR 6 —, where R 6 is as defined above, (Q) —NR 6 —CO—NR 6 —, where R 6 are the same or different and are as defined above, (R) —NR 6 —CO—O—, where R 6 is as defined above, or (S) are absent where the bonds emanating from (B), (D), (E) and (G)-(R) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, and not more than two of (B)-(F), three of (G)-(H) or one of (I)-(R) are present, where U is: (A) hydrogen, (B) lower-alkyl, (C) cycloalkyl, (D) cyano, (E) optionally substituted cycloalkyl, (F) optionally substituted aryl, or (G) optionally substituted heterocyclyl; where R 3 is: (I) hydrogen, (II) hydroxy, (III) lower-alkoxy, or (IV) lower-alkenyloxy; where R 4 is: (I) hydrogen, (II) lower-alkyl, (III) lower-alkenyl, (IV) lower-alkoxy, (V) hydroxy-lower-alkyl, (VI) lower-alkoxy-lower-alkyl, (VII) benzyl, (VIII) oxo, or (IX) where R 3 and R 4 together are a bond, or R 4a -Z 1 -X 1 — where R 4 , is (A) H—, (B) lower-alkyl-, (C) lower-alkenyl-, (D) hydroxy-lower-alkyl-, (E) polyhydroxy-lower-alkyl-, (F) lower-alkyl-O-lower-alkyl-, (G) aryl-, (H) heterocyclyl-, (I) arylalkyl-, (J) heterocyclyloxylalkyl-, (K) aryloxyalkyl-, (L) heterocyclyloxylalkyl-, (M) (R 5 R 6 )—N—(CH 2 ) 1-3 —, where R 1 and R 6 are as defined above, (N) (R 5 R 6 )—N—, where R 5 and R 6 are as defined above, (O) lower-alkyl-S(O) 0-2 —, (P) aryl-S(O) 0-2 —, (Q) heterocyclyl-S(O) 0-2 —, (R) HO—SO 3 — or a salt thereof, (S) H 2 N—C(NH)—NH—, or (T) NC—, where the bonds emanating from (N)-(T) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom, where Z 1 is: (A) a bond, (B) lower-alkylene-, (C) lower-alkenylene-, (D) —O—, (E) —N(R 11 )—, where R 11 is hydrogen or lower-alkyl, (F) —S(O) 0-2 —, (G) —CO—, (H) —O—CO—, (I) —O—CO—O—, (J) —O—CO—N(R 11 )O, where R 11 is as defined above, (K) —N(R 11 )—CO—O—, where R 11 is as defined above, (L) —CO—N(R 11 )—, where R 11 is as defined above, (M) —N(R 11 )—CO—, where R 11 is as defined above, (N) —N(R 11 )—CO—N(R 11 )—, where R 11 are the same or different and are as defined above, (O) —CH(OR 9 )—, where R 9 is hydrogen, lower-alkyl, acyl or arylalkyl, or (P) is absent where the bonds emanating from (D) and (H)-(O) join to a C atom of the adjacent group and this C atom is saturated when the bond emanates from a heteroatom; where X 1 is: (A) a bond, (B) —O—, (C) —N—(R 11 )—, where R 11 is as defined above, (D) —S(O) 0-2 —, (E) —(CH 2 ) 1-3 —, or (F) is absent; where Q is: (I) ethylene, or (II) is absent; where X is: (I) a bond, (II) —O—, (III) —S—, (IV) —CH—R 11 —, where R 11 is as defined above, (V) —CHOR 9 —, where R 9 is as defined above, (VI) —O—CO, (VII) —CO—, or (VIII) C═NOR 10 —, where R 10 is carboxyalkyl, alkoxycarbonylalkyl, alkyl or hydrogen, with the bond emanating from an oxygen or sulfur atom joining to a saturated C atom of group Z or to R 1 ; where W is: (I) —O—, or (II) —S—; where Z is: (I) lower-alkylene, (II) lower-alkenylene, (III) hydroxy-lower-alkylidene, (IV) —O—, (V) —S—, (VI) —O-Alk-, where Alk is a lower alkylene (VII) —S-Alk-, where Alk is as defined above, (VIII) -Alk-O—, where Alk is as defined above, or (IX) -Alk-S, where Alk is as defined above; where n is: (I) one, or (II) zero or one when X is —O—CO; and where m is 0 or 1; with the provisos that (I) X is —CH—R 11 — and either R 2 contains a substituent L 1 -T 1 -L 2 -T 2 -L 3 -T 3 -L 4 -T 4 -L 5 -U or R 4 is a substituent defined above other than hydrogen when Z is —O— or —S—, (II) X is —CH—R 11 — when Z is —O-Alk- or —S-Alk-, and (III) Z is lower-alkenylene, -Alk-O— or -Alk-S— when X is a bond, or a pharmaceutically acceptable salt thereof. detailed-description description="Detailed Description" end="lead"? |
Positioning system |
A system to position at least one vessel or object in an aquatic environment including: each vessel or object having a receiver connected thereto; and at least two transmitters; wherein each transmitter transmits a unique signal and the receiver on each vessel or object receives each unique signal and a calculation means determines the position of the vessel or object based on the received signals. |
1. A system to position at least one vessel or object in an aquatic environment including: each said vessel or object having a respective passive receiver connected thereto; at least two transmitters located below the surface of said aquatic environment and raised above the bottom of said aquatic environment; and a location means to determine the position of each said transmitter; wherein each said transmitter transmits a signal that uniquely identifies each said transmitter and is transmitted independent of a command or interrogation signal, and said receiver on each said vessel or object receives each said signal and a calculation means determines the position of said vessel or object based on the received signals. 2. A system as claimed in claim 1, wherein each said transmitter transmits its signal at predetermined times. 3. A system as claimed in claim 1, wherein said transmitter is a transmitting hydrophone. 4. A system as claimed in claim 1, wherein each said transmitter is attached to a respective housing located at the bottom of the aquatic environment. 5. A system as claimed in claim 4, wherein said transmitter is fixed above the floor of said aquatic environment. 6. A system as claimed in claim 4, wherein said transmitter is attached to a floatation means and is floated above said housing. 7. A system as claimed in claim 6, wherein said location means tracks the position of said flotation means. 8. A system as claimed in claim 6, wherein said location means includes a transmitting hydrophone attached to said a housing, and at least three receiving hydrophones attached to said floatation means. 9. A system as claimed in claim 6, wherein said location means includes a transmitting hydrophone attached to said floatation means, and at least three receiving hydrophones located on said housing. 10. A system as claimed in claim 1, wherein at predetermined intervals one of said transmitters will suspend transmission in order to receive a transmission from another said transmitter to enable clock signals of said transmitters to be calibrated or synchronised. 11. A system as claimed in claim 1, wherein said receiver includes a beam former to minimise interference from any noise emanating from said vessel or object to which said receiver is connected. 12. A system as claimed in claim 1, further including a display means for displaying position information. 13. A system as claimed in claim 4, wherein said housing is connected to an external power source. 14. A system as claimed in claim 1 wherein said transmitters are arranged in a hexagonal, square, diamond, octagonal or triangular shaped grid. 15. A system as claimed in claim 1, wherein said transmitters are spaced apart by a distance equal to or greater than the depth of the aquatic environment. 16. A system as claimed in claim 1, wherein said transmitters are located at least 2 km apart. 17. A system as claimed in claim 4, wherein said housing includes: a main controller for controlling components in said housing; a signal processing means for modifying any signals transmitted or received; a power source to provide power to said housing; and a signal generator to generate said unique signal. 18. A system as claimed in claim 1, wherein said receiver includes: at least one receiving hydrophone for receiving said unique signals; a beam former to minimise interference to signals; and a processing means for controlling components in said receiver. 19. A system as claimed in claim 18, wherein said processing means displays the location on a display unit. 20. A system as claimed in claim 1, further including a stationary reference receiver to assist in the locating and positioning of said object. 21. (canceled) |
<SOH> BACKGROUND OF THE INVENTION <EOH>The positioning and location of vessels or objects in an aquatic environment is a necessary requirement in many industries. However, existing underwater and surface acoustic positioning and navigation systems do have a number of problems, including: 1. Only single vessel/user-positioning systems. That is, existing acoustic tracking systems are single user, and accordingly each user has their own set of underwater beacons. 2. Underwater acoustic pollution, causing interference with ocean based animals (e.g. whales) and other acoustic systems. Existing acoustic tracking systems use ping or chirp signalling similar to the acoustic signals used by ocean based animals. Thus from a system view, biological noise can interfere with tracking systems, and further multiple acoustic tracking systems can interfere with each other. 3. Limited range of coverage—due to the physical properties of deepwater on acoustic signals interfering with the transmitted signal (e.g. ray-bending, signal dispersion, vessel noise, etc.). 4. The need, time and cost to re-deploy the sea-floor components (beacons) for each drilling or construction site. As noted above, existing acoustic tracking systems are single user and cover a relatively small area of approximately one square kilometre. Users are required to demobilise their own underwater infrastructure when operations move outside the tracking boundaries, which is an expensive process. Currently available systems being used in the oil and gas and underwater construction industry do not address the problems listed above and in fact most likely contribute to the underwater acoustic pollution. An example system currently used in the positioning of vessels is the long base line system. Existing Long Base Line (LBL) systems use multiple beacons to form up to a 1 km base line. The user positions itself relative to the base line. The position of the beacons must be determined and the position calibration is performed by sailing around a beacon, and range tracking the beacon relative to the differential global positioning system (DGPS) position. The data is then post-processed to determine the beacon position. This system has been the best-known attempt to date to tackle part of the identified problem. However, deploying numerous sea-floor components (say 80) along the path of an area of operation has still been for single vessel use, has significant deployment overhead and has added to the acoustic pollution problem. Communication using digital spread spectrum (DSS) signalling has also been used in this industry for various applications, however the range and reliability of these systems have so far been unable to satisfy the overall problem. This combination of problems has been difficult to solve due to the known signalling techniques (means for transmitting acoustics) being unable to provide a means for communicating over long distances underwater (>2 km) with the required level of reliability and power consumption. |
<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>With the above objectives in mind, the present invention provides a system to position at least one vessel or object in an aquatic environment including: each vessel or object having a respective passive receiver connected thereto; at least two transmitters located below the surface of said aquatic environment and raised above the bottom of said aquatic environment; and a location means to determine the position of each said transmitter; An example system currently used in the positioning of vessels is the long base line system. Existing Long Base Line (LBL) systems use multiple beacons to form up to a 1 km base line. The user positions itself relative to the base line. The position of the beacons must be determined and the position calibration is performed by sailing around a beacon, and range tracking the beacon relative to the differential global positioning system (DGPS) position. The data is then post-processed to determine the beacon position. This system has been the best-known attempt to date to tackle part of the identified problem. However, deploying numerous sea-floor components (say 80) along the path of an area of operation has still been for single vessel use, has significant deployment overhead and has added to the acoustic pollution problem. Communication using digital spread spectrum (DSS) signalling has also been used in this industry for various applications, however the range and reliability of these systems have so far been unable to satisfy the overall problem. This combination of problems has been difficult to solve due to the known signalling techniques (means for transmitting acoustics) being unable to provide a means for communicating over long distances underwater (>2 km) with the required level of reliability and power consumption. |
Method for treatment of cancer and compositions for use therein |
The present invention provides a method for the treatment of a tumor in a subject. The method comprises administering to the subject a composition comprising a therapeutically effective amount of a compound of Formula (I), wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R2 is selected from H, or substituted or unsubstituted alkyl. R3 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, 5- or 6-membered heterocyclic ring the heteroatom(s) of which are selected from one of more of O, S and/or N, —SR14, —OR15, —SOR16, —SO2R17, —SCN, —C(O)—R18, —OR19, NR20COOR21, where R15 to R21 are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl; or an analogue or metabolite thereof. |
1. A method for the treatment of a tumor in a subject, the method comprising administering to the subject a composition comprising a lipid carrier and a therapeutically effective amount of a compound of Formula I: wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R2 is selected from H, or substituted or unsubstituted alkyl. R3 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, 5- or 6-membered heterocyclic ring the heteroatom(s) of which are selected from one of more of O, S and/or N, —SR14, —OR15, —SOR16, —SO2R17, —SCN, —C(O)—R18, —OR19, NR20COOR21, where R15 to R21 are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl; or an analogue or metabolite thereof. 2. A method as claimed in claim 1 in which the R1 substitution occurs in the 5 or 6 position. 3. A method as claimed in claim 2 in which the R1 substitution occurs in the 5 position. 4. A method as claimed in claim 1 in which the compound is benzimidazole carbamate of Formula II: wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R21 is H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl. 5. A method as claimed in claim 1 in which the compound is a compound of Formula III wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or OR12, COOR13, —NO2 NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R21 is H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl. 6. A method as claimed in claim 1 in which the compound is selected from the group consisting of albendazole, albendazole sulphoxide, mebendazole, flubendazole, triclabendazole, oxfenbendazole, luxabendazole, cambendazole, oxibendazole, parbendazole, thiabendazole or fenbendazole. 7. A method as claimed in claim 1 in which the compound is albendazole or an analogue or metabolite thereof. 8. A method as claimed in claim 1 in which the tumour is hepatoma. 9. A method as claimed in claim 1 in which the tumour is selected from the group consisting of colorectal cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer, renal cancer, sarcoma and secondary metastases thereof. 10. A method as claimed in claim 1 in which the compound is co-administered with methimazole. 11. A method of treatment of a tumour in a subject, the method comprising regionally delivering to the site of the tumour by intra-peritoneal, intra-pleural or intra-arterial administration a composition comprising a therapeutically effective amount of a compound of Formula I: wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7. —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R2 is selected from H, or substituted or unsubstituted alkyl. R3 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, 5- or 6-membered heterocyclic ring the heteroatom(s) of which are selected from one of more of O, S and/or N, —SR14, —OR15, —SOR16, —SO2R17, —SCN, —C(O)—R18, —OR19, NR20COOR21, where R15 to R21, are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl; or an analogue or metabolite thereof. 12. A method as claimed in claim 11 in which the R1 substitution occurs in the 5 or 6 position. 13. A method as claimed in claim 12 in which the R1 substitution occurs in the 5 position. 14. A method as claimed in claim 11 in which the compound is benzimidazole carbamate of Formula II: wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, C(O)—R11 or OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R21 is H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl. 15. A method as claimed in claim 11 in which the compound is a compound of Formula III wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R21 is H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl. 16. A method as claimed in claim 11 in which the compound is selected from the group consisting of albendazole, albendazole sulphoxide, mebendazole, flubendazole, triclabendazole, oxfenbendazole, luxabendazole, cambendazole, oxibendazole, parbendazole, thiabendazole or fenbendazole. 17. A method as claimed in claim 11 in which the compound is albendazole or an analogue or metabolite thereof. 18. A method as claimed in claim 11 in which the tumour is hepatoma. 19. A method as claimed in claim 11 in which the tumour is a secondary cancer in the liver. 20. A method as claimed in claim 18 in which the composition is delivered to the liver is via the intrahepatic artery. 21. A method as claimed in claim 11 in which the tumour is selected from the group consisting of colorectal cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer and renal cancer. 22. A method as claimed in claim 11 in which the composition further comprises a pharmaceutically acceptable carrier. 23. A method as claimed in claim 22 in which the carrier is a lipid. 24. A method as claimed in claim 23 in which the lipid is an oil. 25. A method as claimed in claim 22 in which the carrier is an iodised oil. 26. A method as claimed in claim 25 in which the iodised oil is an iodinated ethyl ester of the poppy seed oil. 27. A method as claimed in claim 11 in which the composition further comprises methimazole. 28. A pharmaceutical composition for use in the treatment of a tumour in a subject, the composition comprising a lipid carrier and an effective amount of a compound of Formula I: wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R2 is selected from H, or substituted or unsubstituted alkyl. R3 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, 5- or 6-membered heterocyclic ring the heteroatom(s) of which are selected from one of more of O, S and/or N, —SR14, —OR15, —SOR16, —SO2R17, —SCN, —C(O)—R18, —OR19, NR20COOR21, where R15 to R21 are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl; or an analogue or metabolite thereof. 29. A composition as claimed in claim 28 in which the R1 substitution occurs in the 5 or 6 position. 30. A composition as claimed in claim 29 in which the R1 substitution occurs in the 5 position. 31. A composition as claimed in claim 28 in which the compound is benzimidazole carbamate of Formula II: wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8, —SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is I to 4; R21 is H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl. 32. A composition as claimed in claim 28 in which the compound is a compound of Formula III wherein R1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR7, —SOR8—SO2R9, —SCN, B′(CH2)nBR10, —C(O)—R11 or —OR12, COOR13, —NO2, NR13aCOOR13b, isothiocyanato, or —CN where R7 to R13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO2 and n is 1 to 4; R21 is H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl. 33. A composition as claimed in claim 28 in which the compound is selected from the group consisting of albendazole, albendazole sulphoxide, mebendazole, flubendazole, triclabendazole, oxfenbendazole, luxabendazole, cambendazole, oxibendazole, parbendazole, thiabendazole or fenbendazole. 34. A composition as claimed in claim 28 in which the compound is albendazole or an analogue or metabolite thereof. 35. A composition as claimed in claim 28 in which the carrier is an oil. 36. A composition as claimed in claim 35 in which the carrier is an iodised oil. 37. A composition as claimed in claim 36 in which the iodised oil is an iodinated ethyl ester of the poppy seed oil. 38. A composition as claimed in claim 28 in which the composition further comprises methimazole. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Hepatocellular carcinoma (HCC; hepatoma) is one of the most common malignancies and a leading cause of death worldwide (1-3). Untreated, HCC typically has a dismal prognosis. Surgical resection remains the mainstay for treatment of HCC and provides the only consistent long-term tumor-free survival (4). However, resection has been limited primarily by low resectibility rates and recurrent disease. Systemic chemotherapy as a primary treatment modality for HCC has limited value because only a small portion of patients will obtain meaningful palliation with the presently available drugs and regimens (2, 4, 5,) and because the toxicity of currently available chemotherapeutic agents often outweighs their limited benefits (6). Furthermore, liver is the most common site for metastases of colorectal carcinoma which in itself is the leading cause of cancerous death in non-smokers in the developed world (7). Albendazole (ABZ; methyl 5-propylthio-1H-benzimidazole-2-yl carbamate) is a benzimidazole carbamate (BZs) anthelmintic developed as a veterinary product in 1975. The BZs are now important broad-spectrum drugs for the control of helminth parasites in mammals. They are effective against lungworms and gastrointestinal nematodes, tapeworms and liver flukes (8). The intrinsic anthelmintic action of benzimidazole compounds on parasite relies on a progressive disruption of basic cell functions as a result of their binding to parasite tubulin and depolymerization of microtubules. However, a number of other mechanisms including disruption of glucose uptake and metabolism have also been described for these compounds (9-11). |
<SOH> SUMMARY OF THE INVENTION <EOH>The present inventors tested BZs and particularly albendazole against a range of liver (HepG2, Hep3B, PLC/PRF/5, SKHEP-1, Hep1-6, HTC, Novikoff) and colorectal cancer (C-170, HT-29 and LOVO) cell lines. The results obtained show potent and dose dependent inhibition of proliferation of these cells by albendazole (and several other BZs). Albendazole was effective against all human and animal cell lines examined, and over a 5 day treatment period, [3H]thymidine incorporation was reduced by over 80% (range 81.6-99.4%) in all these cell lines. Accordingly, in a first aspect, the present invention provides a method for the treatment of a tumor in a subject, the method comprising administering to the subject a composition comprising a therapeutically effective amount of a compound of Formula I: wherein R 1 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, —SR 7 , —SOR 8 , —SO 2 R 9 , —SCN, B′(CH 2 ) n BR 10 , —C(O)—R 11 or —OR 12 , COOR 13 —NO 2 , NR 13a COOR 13b , isothiocyanato, or —CN where R 7 to R 13b are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, B and B′ are independently selected from O, S, S(O) or SO 2 and n is 1 to 4; R 2 is selected from H, or substituted or unsubstituted alkyl. R 3 is selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl, 5- or 6-membered heterocyclic ring the heteroatom(s) of which are selected from one of more of O, S and/or N, —SR 14 , —OR 15 , —SOR 16 , —SO 2 R 17 , —SCN, —C(O)—R 18 , —OR 19 , NR 20 COOR 21 , where R 15 , to R 21 are each independently selected from H, substituted or unsubstituted, straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or arylalkyl; or an analogue or metabolite thereof. |
Electric connector |
An electric connector (2, 2′) having an insulating casing (4) defining a number of cavities for housing respective electric terminals and having axes parallel to a first direction (A) in which the connector (2, 2′) is coupled to a complementary connector (3); a slide (16) fitted to the casing (4) to slide in a second direction (B) perpendicular to the first direction (A), and having cam-type first engaging members (22) for receiving respective second engaging members (23) on the complementary connector (3) to produce a relative coupling movement of the connectors (2, 2′; 3) in the first direction (A) when the slide (16) is moved in the second direction (B) into a fully assembled position with respect to the casing (4); and retaining means (40, 41, 62) for keeping the slide (16) partly connected to the casing (4); the retaining means (40, 41, 62) being selectively deactivated when coupling the connector (2, 2′) to the complementary connector (3). |
1) An electric connector (2,2′) comprising an insulating casing (4) defining a number of cavities for housing respective electric terminals and having axes parallel to a first direction (a) in which said connector (2,2′) is coupled to a complementary connector (3); a slide (16) fitted to said casing (4) to slide in a second direction (b) perpendicular to said first direction (a), and having cam-type first engaging members (22) for receiving respective second engaging members (23) on said complementary connector (3) to produce a relative coupling movement of said connectors (2,2′; 3) in said first direction (a) when said slide (16) is moved in said second direction (b) into a fully assembled position with respect to said casing (4); and releasable constraint means (40,41,62) for keeping said slide (16) partly connected to said casing (4); characterized in that said releasable constraint means comprise retaining means (40, 41,62) for locking said slide (16) to said casing (4); said retaining means (40,41,62) being selectively deactivated when coupling said connector (2,2′) to said complementary connector (3). 2) A connector as claimed in claim 1, characterized by comprising an actuating lever (30) connected movably to said casing (4) to move said slide (16) into said fully assembled position. 3) A connector as claimed in claim 2, characterized in that said lever (30) comprises at least one hinge portion (32) hinging it to said casing (4) about an axis (C) perpendicular to said first and said second direction (A, B); and in that said hinge portion (32) comprises, with reference to said axis (C), an angular coupling portion (35) for engaging said slide (16), and a free angular portion (63) for releasing the slide (16). 4) A connector as claimed in claim 3, characterized in that said coupling portion of said hinge portion (32) of said lever (30) comprises a sector gear (35) meshing with a rack (37) on said slide (16); and in that said free portion (63) of said hinge portion (32) of said lever (30) is movable clear of the profile of said rack (37) on said slide (16). 5) A connector as claimed in claim 2, characterized in that said retaining means comprise releasable one-way stop means (62) acting on said slide (16) to prevent the slide (16) from moving into said fully assembled position, and defined by said lever (30) in a deactivated position. 6) A connector as claimed in claim 5, characterized in that said stop means comprise a stop tooth (62) projecting radially from said hinge portion (32) of said lever (30), and defining a stop surface for said rack (37) on said slide (16) when said lever (30) is in said deactivated position; and in that said free portion (63) of said hinge portion (32) of said lever (30) is defined on opposite sides by said sector gear (35) and said stop tooth (62). 7) A connector as claimed in claim 4, characterized in that said sector gear (35) meshes with said rack (37) as said lever (30) is rotated between a first and a second operating position corresponding respectively to a first partly assembled position of said slide (16) with respect to said casing (4) and to said fully assembled position; and in that said free portion (63) passes clear of said rack (37) as said lever (30) is rotated between said first operating position and said deactivated position. 8) A connector as claimed in claim 7, characterized by comprising rigid connecting means (66,67,78) for rigidly connecting said lever (30) to said casing (4) in said deactivated position; said rigid connecting means (66,67,78) being selectively releasable to move said lever (30) into said first operating position. 9) A connector as claimed in claim 8, characterized in that said rigid connecting means comprise fastening means (66,67,78) for fastening said lever [(30)] to said casing (4). 10) A connector as claimed in claim 1, characterized in that said retaining means comprise elastically flexible one-way locking means (40) which interfere with the sliding movement of said slide (16) into said fully assembled position, and which are set by correct engagement of said first and said second engaging members (22,23) to a deformed, configuration allowing said slide (16) to move into said fully assembled position. 11) A connector as claimed in claim 10, characterized in that said first engaging members comprise a number of cam grooves (22) formed on said slide (16); and in that said locking means (40) extend through at least one of said grooves (22), and are set to said deformed configuration by interacting with a relative one of said second engaging members (23) on said complementary connector (3) engaging said one of said grooves (22). 12) A connector as claimed in claim 11, characterized in that said locking means (40) comprise at least one elastically flexible member (42,71), which is carried by one of said casing (4) and said slide (16), has an interference portion (43,75) interfering with the other of said casing (4) and said slide (16), and is activated by a relative one of said second engaging members (23) on said complementary connector (3) engaging said one of said grooves (22). 13) A connector as claimed in claim 12, characterized in that said elastically flexible member (42) projects from a wall (11) of said casing (4), and comprises an end tooth (43) defining said interference portion, and which engages an inlet portion (25) of said one of said grooves (22), and is released from said inlet portion (25) by interacting with the relative one of said second engaging members (23) engaging said one of said grooves (22). 14) A connector as claimed in claim 12, characterized in that said elastically flexible member (71) projects from said slide (16), cooperates with an interacting portion (72) of said casing (4) to prevent said slide (16) from moving into said fully assembled position, and is releasable from said interacting portion (72) in said deformed configuration. 15) A connector as claimed in claim 14, characterized in that said interacting portion is defined by a through opening (72) formed in said casing (4) and engaged by said elastically flexible member (71). 16) A connector as claimed in claim 15, characterized in that said elastically flexible member (71) comprises an intermediate portion (73) connected to said slide (16) by an elastic hinge (74); a first end portion (75) engaging said opening (72) in said casing (4); and an opposite second end portion (76) which projects through an inlet portion (25) of said one of said grooves (22). 17) A connector as claimed in claim 10, characterized in that said retaining means comprise second releasable one-way locking means (41) exerting on said slide (16) retaining forces opposite those exerted by said stop means (62) and said locking means (40), to prevent withdrawal of the slide (16) from said casing (4). 18) A connector as claimed in claim 17, characterized in that said second locking means (41) can be set to a first and a second operating configuration; said second locking means (41), in said first operating configuration, and said locking means (40) temporarily retaining said slide (16) in said first partly assembled position and said lever (30) in said first operating position; said second locking means (41), in said second operating configuration, and said stop means (62) temporarily retaining said slide (16) in a second partly assembled position with respect to said casing (4) corresponding to said deactivated position of said lever (30); and said first partly assembled position of said slide (16) being a position interposed, in said second direction (B), between said second partly assembled position and said fully assembled position. 19) A connector as claimed in claim 18, characterized in that said second locking means comprise at least one elastic lance (52) carried by one of said casing (4) and said slide (16); and two seats (56,68) formed on the other of said casing (4) and said slide (16), and which are located successively in said second direction (B), and are engaged by said lance (52) to respectively define said first and second partly assembled position of said slide (16). 20) A connector as claimed in claim 1, characterized in that said retaining means (40, 41, 62) define a first and a second partly assembled position of said slide (16) with respect to said casing (4); said first partly assembled position of said slide (16) being a position interposed, in said second direction (B), between said second partly assembled position and said fully assembled position. |
<SOH> BACKGROUND ART <EOH>Connecting units of the above type are known in which the connectors comprise respective insulating casings defining respective numbers of cavities for housing respective connectable male and female electric terminals. Such units normally comprise a lever-and-slide coupling device, which is operated manually when the plug and socket connectors are engaged to couple the connectors with a minimum amount of effort. The lever-and-slide coupling device substantially comprises a slide fitted to slide inside the plug connector casing in a direction perpendicular to the coupling direction of the connectors; and an actuating lever hinged to the plug connector casing and connected to the slide. In a fairly common embodiment, the slide is C-shaped and defined by an end wall perpendicular to the sliding direction, and by two lateral walls extending perpendicularly from respective opposite end edges of the end wall, and which slide along respective lateral walls of the plug connector casing. Each lateral wall of the slide has a number of cam grooves for receiving respective external pins on the socket connector, and for producing a relative engaging movement of the plug and socket connectors in the coupling direction when the slide is moved in the sliding direction. The slide is normally retained, by releasable retaining means, e.g. click-on retaining members, in a preassembly position partly inserted inside the plug connector casing, and is moved into a fully inserted position inside the casing by rotating the actuating lever from a raised to a lowered position about its hinge axis. To function properly, the releasable retaining means must be sized and designed to ensure a given load by which to retain the slide inside the casing. The load, however, may not be sufficient to prevent the slide from being inserted accidentally inside the plug connector casing, in the event the plug connector is knocked, dropped, etc. before being coupled to the complementary connector. In which case, the slide must be reset to the preassembly position before the connectors are coupled, thus complicating assembly of the connecting unit. By way of a solution to the problem, the load exerted by the retaining means could be increased, though this would also mean a corresponding increase in the force required on the actuating lever to couple the connectors, thus impairing the function for which the lever-and-slide coupling device was designed, i.e. to permit coupling of the connectors with a minimum amount of effort. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>Two preferred, non-limiting embodiments of the present invention will be described by way of example with reference to the accompanying drawings, in which: FIG. 1 shows an exploded view in perspective, with parts removed for clarity, of an electric connecting unit defined by an electric plug connector in accordance with the present invention, and by a complementary electric socket connector; FIG. 2 shows a larger-scale view in perspective of a detail of the FIG. 1 electric plug connector; FIG. 3 shows a larger-scale view in perspective of a further detail of the FIG. 1 electric plug connector; FIG. 4 shows a larger-scale view in perspective of a slide of the FIG. 1 plug connector; FIG. 5 shows a larger-scale side view of the FIG. 1 electric plug connector in a different configuration; FIG. 6 shows a larger-scale view in perspective of the FIG. 3 detail of the electric plug connector in the FIG. 5 configuration; FIG. 7 shows a larger-scale section along line VII-VII in FIG. 5 ; FIG. 8 shows a side view of a further embodiment of an electric plug connector in accordance with the present invention; FIG. 9 shows a larger-scale view in perspective of a detail of the FIG. 8 electric plug connector; FIG. 10 shows a cross section in perspective of the FIG. 9 detail. detailed-description description="Detailed Description" end="lead"? |
Streptococcus pneumoniae proteins and nucleic acids |
The invention provides proteins and nucleic acid sequences from Streptococcus pneumoniae, together with a genome sequence. These are useful for the development of vaccines, diagnostics, and antibiotics. |
1. A protein comprising an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130,132,134,136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 690, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, 776, 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, 892, 894, 896, 898, 900, 902, 904, 906, 908, 910, 912, 914, 916, 918, 920, 922, 924, 926, 928, 930, 932, 934, 936, 938, 940, 942, 944, 946, 948, 950, 952, 954, 956, 958, 960, 962, 964, 966, 968, 970, 972, 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1016, 1018, 1020, 1022, 1024, 1026, 1028, 1030, 1032, 1034, 1036, 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1062, 1064, 1066, 1068, 1070, 1072, 1074, 1076, 1078, 1080, 1082, 1084, 1086, 1088, 1090, 1092, 1094, 1096, 1098, 1100, 1102, 1104, 1106, 1108, 1110, 1112, 1114, 1116, 1118, 1120, 1122, 1124, 1126, 1128, 1130, 1132, 1134, 1136, 1138, 1140, 1142, 1144, 1146, 1148, 1150, 1152, 1154, 1156, 1158, 1160, 1162, 1164, 1166, 1168, 1170, 1172, 1174, 1176, 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 1238, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310, 1312, 1314, 1316, 1318, 1320, 1322, 1324, 1326, 1328, 1330, 1332, 1334, 1336, 1338, 1340, 1342, 1344, 1346, 1348, 1350, 1352, 1354, 1356, 1358, 1360, 1362, 1364, 1366, 1368, 1370, 1372, 1374, 1376, 1378, 1380, 1382, 1384, 1386, 1388, 1390, 1392, 1394, 1396, 1398, 1400, 1402, 1404, 1406, 1408, 1410, 1412, 1414, 1416, 1418, 1420, 1422, 1424, 1426, 1428, 1430, 1432, 1434, 1436, 1438, 1440, 1442, 1444, 1446, 1448, 1450, 1452, 1454, 1456, 1458, 1460, 1462, 1464, 1466, 1468, 1470, 1472, 1474, 1476, 1478, 1480; 1482, 1484, 1486, 1488, 1490, 1492, 1494, 1496, 1498, 1500, 1502, 1504, 1506, 1508, 1510, 1512, 1514, 1516, 1518, 1520, 1522, 1524, 1526, 1528, 1530, 1532, 1534, 1536, 1538, 1540, 1542, 1544, 1546, 1548, 1550, 1552, 1554, 1556, 1558, 1560, 1562, 1564, 1566, 1568, 1570, 1572, 1574, 1576, 1578, 1580, 1582, 1584, 1586, 1588, 1590, 1592, 1594, 1596, 1598, 1600, 1602, 1604, 1606, 1608, 1610, 1612, 1614, 1616, 1618, 1620, 1622, 1624, 1626, 1628, 1630, 1632, 1634, 1636, 1638, 1640, 1642, 1644, 1646, 1648, 1650, 1652, 1654, 1656, 1658, 1660, 1662, 1664, 1666, 1668, 1670, 1672, 1674, 1676, 1678, 1680, 1682, 1684, 1686, 1688, 1690, 1692, 1694, 1696, 1698, 1700, 1702, 1704, 1706, 1708, 1710, 1712, 1714, 1716, 1718, 1720, 1722, 1724, 1726, 1728, 1730, 1732, 1734, 1736, 1738, 1740, 1742, 1744, 1746, 1748, 1750, 1752, 1754, 1756, 1758, 1760, 1762, 1764, 1766, 1768, 1770, 1772, 1774, 1776, 1778, 1780, 1782, 1784, 1786, 1788, 1790, 1792, 1794, 1796, 1798, 1800, 1802, 1804, 1806, 1808, 1810, 1812, 1814, 1816, 1818, 1820, 1822, 1824, 1826, 1828, 1830, 1832, 1834, 1836, 1838, 1840, 1842, 1844, 1846, 1848, 1850, 1852, 1854, 1856, 1858, 1860, 1862, 1864, 1866, 1868, 1870, 1872, 1874, 1876, 1878, 1880, 1882, 1884, 1886, 1888, 1890, 1892, 1894, 1896, 1898, 1900, 1902, 1904, 1906, 1908, 1910, 1912, 1914, 1916, 1918, 1920, 1922, 1924, 1926, 1928, 1930, 1932, 1934, 1936, 1938, 1940, 1942, 1944, 1946, 1948, 1950, 1952, 1954, 1956, 1958, 1960, 1962, 1964, 1966, 1968, 1970, 1972, 1974, 1976, 1978, 1980, 1982, 1984, 1986, 1988, 1990, 1992, 1994, 1996, 1998, 2000, 2002, 2004, 2006, 2008, 2010, 2012, 2014, 2016, 2018, 2020, 2022, 2024, 2026, 2028, 2030, 2032, 2034, 2036, 2038, 2040, 2042, 2044, 2046, 2048, 2050, 2052, 2054, 2056, 2058, 2060, 2062, 2064, 2066, 2068, 2070, 2072, 2074, 2076, 2078, 2080, 2082, 2084, 2086, 2088, 2090, 2092, 2094, 2096, 2098, 2100, 2102, 2104, 2106, 2108, 2110, 2112, 2114, 2116, 2118, 2120, 2122, 2124, 2126, 2128, 2130, 2132, 2134, 2136, 2138, 2140, 2142, 2144, 2146, 2148, 2150, 2152, 2154, 2156, 2158, 2160, 2162, 2164, 2166, 2168, 2170, 2172, 2174, 2176, 2178, 2180, 2182, 2184, 2186, 2188, 2190, 2192, 2194, 2196, 2198, 2200, 2202, 2204, 2206, 2208, 2210, 2212, 2214, 2216, 2218, 2220, 2222, 2224, 2226, 2228, 2230, 2232, 2234, 2236, 2238, 2240, 2242, 2244, 2246, 2248, 2250, 2252, 2254, 2256, 2258, 2260, 2262, 2264, 2266, 2268, 2270, 2272, 2274, 2276, 2278, 2280, 2282, 2284, 2286, 2288, 2290, 2292, 2294, 2296, 2298, 2300, 2302, 2304, 2306, 2308, 2310, 2312, 2314, 2316, 2318, 2320, 2322, 2324, 2326, 2328, 2330, 2332, 2334, 2336, 2338, 2340, 2342, 2344, 2346, 2348, 2350, 2352, 2354, 2356, 2358, 2360, 2362, 2364, 2366, 2368, 2370, 2372, 2374, 2376, 2378, 2380, 2382, 2384, 2386, 2388, 2390, 2392, 2394, 2396, 2398, 2400, 2402, 2404, 2406, 2408, 2410, 2412, 2414, 2416, 2418, 2420, 2422, 2424, 2426, 2428, 2430, 2432, 2434, 2436, 2438, 2440, 2442, 2444, 2446, 2448, 2450, 2452, 2454, 2456, 2458, 2460, 2462, 2464, 2466, 2468, 2470, 2472, 2474, 2476, 2478, 2480, 2482, 2484, 2486, 2488, 2490, 2492, 2494, 2496, 2498, 2500, 2502, 2504, 2506, 2508, 2510, 2512, 2514, 2516, 2518, 2520, 2522, 2524, 2526, 2528, 2530, 2532, 2534, 2536, 2538, 2540, 2542, 2544, 2546, 2548, 2550, 2552, 2554, 2556, 2558, 2560, 2562, 2564, 2566, 2568, 2570, 2572, 2574, 2576, 2578, 2580, 2582, 2584, 2586, 2588, 2590, 2592, 2594, 2596, 2598, 2600, 2602, 2604, 2606, 2608, 2610, 2612, 2614, 2616, 2618, 2620, 2622, 2624, 2626, 2628, 2630, 2632, 2634, 2636, 2638, 2640, 2642, 2644, 2646, 2648, 2650, 2652, 2654, 2656, 2658, 2660, 2662, 2664, 2666, 2668, 2670, 2672, 2674, 2676, 2678, 2680, 2682, 2684, 2686, 2688, 2690, 2692, 2694, 2696, 2698, 2700, 2702, 2704, 2706, 2708, 2710, 2712, 2714, 2716, 2718, 2720, 2722, 2724, 2726, 2728, 2730, 2732, 2734, 2736, 2738, 2740, 2742, 2744, 2746, 2748, 2750, 2752, 2754, 2756, 2758, 2760, 2762, 2764, 2766, 2768, 2770, 2772, 2774, 2776, 2778, 2780, 2782, 2784, 2786, 2788, 2790, 2792, 2794, 2796, 2798, 2800, 2802, 2804, 2806, 2808, 2810, 2812, 2814, 2816, 2818, 2820, 2822, 2824, 2826, 2828, 2830, 2832, 2834, 2836, 2838, 2840, 2842, 2844, 2846, 2848, 2850, 2852, 2854, 2856, 2858, 2860, 2862, 2864, 2866, 2868, 2870, 2872, 2874, 2876, 2878, 2880, 2882, 2884, 2886, 2888, 2890, 2892, 2894, 2896, 2898, 2900, 2902, 2904, 2906, 2908, 2910, 2912, 2914, 2916, 2918, 2920, 2922, 2924, 2926, 2928, 2930, 2932, 2934, 2936, 2938, 2940, 2942, 2944, 2946, 2948, 2950, 2952, 2954, 2956, 2958, 2960, 2962, 2964, 2966, 2968, 2970, 2972, 2974, 2976, 2978, 2980, 2982, 2984, 2986, 2988, 2990, 2992, 2994, 2996, 2998, 3000, 3002, 3004, 3006, 3008, 3010, 3012, 3014, 3016, 3018, 3020, 3022, 3024, 3026, 3028, 3030, 3032, 3034, 3036, 3038, 3040, 3042, 3044, 3046, 3048, 3050, 3052, 3054, 3056, 3058, 3060, 3062, 3064, 3066, 3068, 3070, 3072, 3074, 3076, 3078, 3080, 3082, 3084, 3086, 3088, 3090, 3092, 3094, 3096, 3098, 3100, 3102, 3104, 3106, 3108, 3110, 3112, 3114, 3116, 3118, 3120, 3122, 3124, 3126, 3128, 3130, 3132, 3134, 3136, 3138, 3140, 3142, 3144, 3146, 3148, 3150, 3152, 3154, 3156, 3158, 3160, 3162, 3164, 3166, 3168, 3170, 3172, 3174, 3176, 3178, 3180, 3182, 3184, 3186, 3188, 3190, 3192, 3194, 3196, 3198, 3200, 3202, 3204, 3206, 3208, 3210, 3212, 3214, 3216, 3218, 3220, 3222, 3224, 3226, 3228, 3230, 3232, 3234, 3236, 3238, 3240, 3242, 3244, 3246, 3248, 3250, 3252, 3254, 3256, 3258, 3260, 3262, 3264, 3266, 3268, 3270, 3272, 3274, 3276, 3278, 3280, 3282, 3284, 3286, 3288, 3290, 3292, 3294, 3296, 3298, 3300, 3302, 3304, 3306, 3308, 3310, 3312, 3314, 3316, 3318, 3320, 3322, 3324, 3326, 3328, 3330, 3332, 3334, 3336, 3338, 3340, 3342, 3344, 3346, 3348, 3350, 3352, 3354, 3356, 3358, 3360, 3362, 3364, 3366, 3368, 3370, 3372, 3374, 3376, 3378, 3380, 3382, 3384, 3386, 3388, 3390, 3392, 3394, 3396, 3398, 3400, 3402, 3404, 3406, 3408, 3410, 3412, 3414, 3416, 3418, 3420, 3422, 3424, 3426, 3428, 3430, 3432, 3434, 3436, 3438, 3440, 3442, 3444, 3446, 3448, 3450, 3452, 3454, 3456, 3458, 3460, 3462, 3464, 3466, 3468, 3470, 3472, 3474, 3476, 3478, 3480, 3482, 3484, 3486, 3488, 3490, 3492, 3494, 3496, 3498, 3500, 3502, 3504, 3506, 3508, 3510, 3512, 3514, 3516, 3518, 3520, 3522, 3524, 3526, 3528, 3530, 3532, 3534, 3536, 3538, 3540, 3542, 3544, 3546, 3548, 3550, 3552, 3554, 3556, 3558, 3560, 3562, 3564, 3566, 3568, 3570, 3572, 3574, 3576, 3578, 3580, 3582, 3584, 3586, 3588, 3590, 3592, 3594, 3596, 3598, 3600, 3602, 3604, 3606, 3608, 3610, 3612, 3614, 3616, 3618, 3620, 3622, 3624, 3626, 3628, 3630, 3632, 3634, 3636, 3638, 3640, 3642, 3644, 3646, 3648, 3650, 3652, 3654, 3656, 3658, 3660, 3662, 3664, 3666, 3668, 3670, 3672, 3674, 3676, 3678, 3680, 3682, 3684, 3686, 3688, 3690, 3692, 3694, 3696, 3698, 3700, 3702, 3704, 3706, 3708, 3710, 3712, 3714, 3716, 3718, 3720, 3722, 3724, 3726, 3728, 3730, 3732, 3734, 3736, 3738, 3740, 3742, 3744, 3746, 3748, 3750, 3752, 3754, 3756, 3758, 3760, 3762, 3764, 3766, 3768, 3770, 3772, 3774, 3776, 3778, 3780, 3782, 3784, 3786, 3788, 3790, 3792, 3794, 3796, 3798, 3800, 3802, 3804, 3806, 3808, 3810, 3812, 3814, 3816, 3818, 3820, 3822, 3824, 3826, 3828, 3830, 3832, 3834, 3836, 3838, 3840, 3842, 3844, 3846, 3848, 3850, 3852, 3854, 3856, 3858, 3860, 3862, 3864, 3866, 3868, 3870, 3872, 3874, 3876, 3878, 3880, 3882, 3884, 3886, 3888, 3890, 3892, 3894, 3896, 3898, 3900, 3902, 3904, 3906, 3908, 3910, 3912, 3914, 3916, 3918, 3920, 3922, 3924, 3926, 3928, 3930, 3932, 3934, 3936, 3938, 3940, 3942, 3944, 3946, 3948, 3950, 3952, 3954, 3956, 3958, 3960, 3962, 3964, 3966, 3968, 3970, 3972, 3974, 3976, 3978, 3980, 3982, 3984, 3986, 3988, 3990, 3992, 3994, 3996, 3998, 4000, 4002, 4004, 4006, 4008, 4010, 4012, 4014, 4016, 4018, 4020, 4022, 4024, 4026, 4028, 4030, 4032, 4034, 4036, 4038, 4040, 4042, 4044, 4046, 4048, 4050, 4052, 4054, 4056, 4058, 4060, 4062, 4064, 4066, 4068, 4070, 4072, 4074, 4076, 4078, 4080, 4082, 4084, 4086, 4088, 4090, 4092, 4094, 4096, 4098, 4100, 4102, 4104, 4106, 4108, 4110, 4112, 4114, 4116, 4118, 4120, 4122, 4124, 4126, 4128, 4130, 4132, 4134, 4136, 4138, 4140, 4142, 4144, 4146, 4148, 4150, 4152, 4154, 4156, 4158, 4160, 4162, 4164, 4166, 4168, 4170, 4172, 4174, 4176, 4178, 4180, 4182, 4184, 4186, 4188, 4190, 4192, 4194, 4196, 4198, 4200, 4202, 4204, 4206, 4208, 4210, 4212, 4214, 4216, 4218, 4220, 4222, 4224, 4226, 4228, 4230, 4232, 4234, 4236, 4238, 4240, 4242, 4244, 4246, 4248, 4250, 4252, 4254, 4256, 4258, 4260, 4262, 4264, 4266, 4268, 4270, 4272, 4274, 4276; 4278, 4280, 4282, 4284, 4286, 4288, 4290, 4292, 4294, 4296, 4298, 4300, 4302, 4304, 4306, 4308, 4310, 4312, 4314, 4316, 4318, 4320, 4322, 4324, 4326, 4328, 4330, 4332, 4334, 4336, 4338, 4340, 4342, 4344, 4346, 4348, 4350, 4352, 4354, 4356, 4358, 4360, 4362, 4364, 4366, 4368, 4370, 4372, 4374, 4376, 4378, 4380, 4382, 4384, 4386, 4388, 4390, 4392, 4394, 4396, 4398, 4400, 4402, 4404, 4406, 4408, 4410, 4412, 4414, 4416, 4418, 4420, 4422, 4424, 4426, 4428, 4430, 4432, 4434, 4436, 4438, 4440, 4442, 4444, 4446, 4448, 4450, 4452, 4454, 4456, 4458, 4460, 4462, 4464, 4466, 4468, 4470, 4472, 4474, 4476, 4478, 4480, 4482, 4484, 4486, 4488, 4490, 4492, 4494, 4496, 4498, 4500, 4502, 4504, 4506, 4508, 4510, 4512, 4514, 4516, 4518, 4520, 4522, 4524, 4526, 4528, 4530, 4532, 4534, 4536, 4538, 4540, 4542, 4544, 4546, 4548, 4550, 4552, 4554, 4556, 4558, 4560, 4562, 4564, 4566, 4568, 4570, 4572, 4574, 4576, 4578, 4580, 4582, 4584, 4586, 4588, 4590, 4592, 4594, 4596, 4598, 4600, 4602, 4604, 4606, 4608, 4610, 4612, 4614, 4616, 4618, 4620, 4622, 4624, 4626, 4628, 4630, 4632, 4634, 4636, 4638, 4640, 4642, 4644, 4646, 4648, 4650, 4652, 4654, 4656, 4658, 4660, 4662, 4664, 4666, 4668, 4670, 4672, 4674, 4676, 4678, 4680, 4682, 4684, 4686, 4688, 4690, 4692, 4694, 4696, 4698, 4700, 4702, 4704, 4706, 4708, 4710, 4712, 4714, 4716, 4718, 4720, 4722, 4724, 4726, 4728, 4730, 4732, 4734, 4736, 4738, 4740, 4742, 4744, 4746, 4748, 4750, 4752, 4754, 4756, 4758, 4760, 4762, 4764, 4766, 4768, 4770, 4772, 4774, 4776, 4778, 4780, 4782, 4784, 4786, 4788, 4790, 4792, 4794, 4796, 4798, 4800, 4802, 4804, 4806, 4808, 4810, 4812, 4814, 4816, 4818, 4820, 4822, 4824, 4826, 4828, 4830, 4832, 4834, 4836, 4838, 4840, 4842, 4844, 4846, 4848, 4850, 4852, 4854, 4856, 4858, 4860, 4862, 4864, 4866, 4868, 4870, 4872, 4874, 4876, 4878, 4880, 4882, 4884, 4886, 4888, 4890, 4892, 4894, 4896, 4898, 4900, 4902, 4904, 4906, 4908, 4910, 4912, 4914, 4916, 4918, 4920, 4922, 4924, 4926, 4928, 4930, 4932, 4934, 4936, 4938, 4940, 4942, 4944, 4946, 4948, 4950, 4952, 4954, 4956, 4958, 4960, 4962, 4964, 4966, 4968, 4970, 4972, 4974, 4976, 4978. 2. A protein having 50% or greater sequence identity to a protein according to claim 1. 3. A protein comprising a fragment of an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 690, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, 776, 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, 892, 894, 896, 898, 900, 902, 904, 906, 908, 910, 912, 914, 916, 918, 920, 922, 924, 926, 928, 930, 932, 934, 936, 938, 940, 942, 944, 946, 948, 950, 952, 954, 956, 958, 960, 962, 964, 966, 968, 970, 972, 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1016, 1018, 1020, 1022, 1024, 1026, 1028, 1030, 1032, 1034, 1036, 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1062, 1064, 1066, 1068, 1070, 1072, 1074, 1076, 1078, 1080, 1082, 1084, 1086, 1088, 1090, 1092, 1094, 1096, 1098, 1100, 1102, 1104, 1106, 1108, 1110, 1112, 1114, 1116, 1118, 1120, 1122, 1124, 1126, 1128, 1130, 1132, 1134, 1136, 1138, 1140, 1142, 1144, 1146, 1148, 1150, 1152, 1154, 1156, 1158, 1160, 1162, 1164, 1166, 1168, 1170, 1172, 1174, 1176, 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 1238, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262; 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310, 1312, 1314, 1316, 1318, 1320, 1322, 1324, 1326, 1328, 1330, 1332, 1334, 1336, 1338, 1340, 1342, 1344, 1346, 1348, 1350, 1352, 1354, 1356, 1358, 1360, 1362, 1364, 1366, 1368, 1370, 1372, 1374, 1376, 1378, 1380, 1382, 1384, 1386, 1388, 1390, 1392, 1394, 1396, 1398; 1400, 1402, 1404, 1406, 1408, 1410, 1412, 1414, 1416, 1418, 1420, 1422, 1424, 1426, 1428, 1430, 1432, 1434, 1436, 1438, 1440, 1442, 1444, 1446, 1448, 1450, 1452, 1454, 1456, 1458, 1460, 1462, 1464, 1466, 1468, 1470, 1472, 1474, 1476, 1478, 1480, 1482, 1484, 1486, 1488, 1490, 1492, 1494, 1496, 1498, 1500, 1502, 1504, 1506, 1508, 1510, 1512, 1514, 1516, 1518, 1520, 1522, 1524, 1526, 1528, 1530, 1532, 1534, 1536, 1538, 1540, 1542, 1544, 1546, 1548, 1550, 1552, 1554, 1556, 1558, 1560, 1562, 1564, 1566, 1568, 1570, 1572, 1574, 1576, 1578, 1580, 1582, 1584, 1586, 1588, 1590, 1592, 1594, 1596, 1598, 1600, 1602, 1604, 1606, 1608, 1610, 1612, 1614, 1616, 1618, 1620, 1622, 1624, 1626, 1628, 1630, 1632, 1634, 1636, 1638, 1640, 1642, 1644, 1646, 1648, 1650, 1652, 1654, 1656, 1658, 1660, 1662, 1664, 1666, 1668, 1670, 1672, 1674, 1676, 1678, 1680, 1682, 1684, 1686, 1688, 1690, 1692, 1694, 1696, 1698, 1700, 1702, 1704, 1706, 1708, 1710, 1712, 1714, 1716, 1718, 1720, 1722, 1724, 1726, 1728, 1730, 1732, 1734, 1736, 1738, 1740, 1742, 1744, 1746, 1748, 1750, 1752, 1754, 1756, 1758, 1760, 1762, 1764, 1766, 1768, 1770, 1772, 1774, 1776, 1778, 1780, 1782, 1784, 1786, 1788, 1790, 1792, 1794, 1796, 1798, 1800, 1802, 1804, 1806, 1808, 1810, 1812, 1814, 1816, 1818, 1820, 1822, 1824, 1826, 1828, 1830, 1832, 1834, 1836, 1838, 1840, 1842, 1844, 1846, 1848, 1850, 1852, 1854, 1856, 1858, 1860, 1862, 1864, 1866, 1868, 1870, 1872, 1874, 1876, 1878, 1880, 1882, 1884, 1886, 1888, 1890, 1892, 1894, 1896, 1898, 1900, 1902, 1904, 1906, 1908, 1910, 1912, 1914, 1916, 1918, 1920, 1922, 1924, 1926, 1928, 1930, 1932, 1934, 1936, 1938, 1940, 1942, 1944, 1946, 1948, 1950, 1952, 1954, 1956, 1958, 1960, 1962, 1964, 1966, 1968, 1970, 1972, 1974, 1976, 1978, 1980, 1982, 1984, 1986, 1988, 1990, 1992, 1994, 1996, 1998, 2000, 2002, 2004, 2006, 2008, 2010, 2012, 2014, 2016, 2018, 2020, 2022, 2024, 2026, 2028, 2030, 2032, 2034, 2036, 2038, 2040, 2042, 2044, 2046, 2048, 2050, 2052, 2054, 2056, 2058, 2060, 2062, 2064, 2066, 2068, 2070, 2072, 2074, 2076, 2078, 2080, 2082, 2084, 2086, 2088, 2090, 2092, 2094, 2096, 2098, 2100, 2102, 2104, 2106, 2108, 2110, 2112, 2114, 2116, 2118, 2120, 2122, 2124, 2126, 2128, 2130, 2132, 2134, 2136, 2138, 2140, 2142, 2144, 2146, 2148, 2150, 2152, 2154, 2156, 2158, 2160, 2162, 2164, 2166, 2168, 2170, 2172, 2174, 2176, 2178, 2180, 2182, 2184, 2186, 2188, 2190, 2192, 2194, 2196, 2198, 2200, 2202, 2204, 2206, 2208, 2210, 2212, 2214, 2216, 2218, 2220, 2222, 2224, 2226, 2228, 2230, 2232, 2234, 2236, 2238, 2240, 2242, 2244, 2246, 2248, 2250, 2252, 2254, 2256, 2258, 2260, 2262, 2264, 2266, 2268, 2270, 2272, 2274, 2276, 2278, 2280, 2282, 2284, 2286, 2288, 2290, 2292, 2294, 2296, 2298, 2300, 2302, 2304, 2306, 2308, 2310, 2312, 2314, 2316, 2318, 2320, 2322, 2324, 2326, 2328, 2330, 2332, 2334, 2336, 2338, 2340, 2342, 2344, 2346, 2348, 2350, 2352, 2354, 2356, 2358, 2360, 2362, 2364, 2366, 2368, 2370, 2372, 2374, 2376, 2378, 2380, 2382, 2384, 2386, 2388, 2390, 2392, 2394, 2396, 2398, 2400, 2402, 2404, 2406, 2408, 2410, 2412, 2414, 2416, 2418, 2420, 2422, 2424, 2426, 2428, 2430, 2432, 2434, 2436, 2438, 2440, 2442, 2444, 2446, 2448, 2450, 2452, 2454, 2456, 2458, 2460, 2462, 2464, 2466, 2468, 2470, 2472, 2474, 2476, 2478, 2480, 2482, 2484, 2486, 2488, 2490, 2492, 2494, 2496, 2498, 2500, 2502, 2504, 2506, 2508, 2510, 2512, 2514, 2516, 2518, 2520, 2522, 2524, 2526, 2528, 2530, 2532, 2534, 2536, 2538, 2540, 2542, 2544, 2546, 2548, 2550, 2552, 2554, 2556, 2558, 2560, 2562, 2564, 2566, 2568, 2570, 2572, 2574, 2576, 2578, 2580, 2582, 2584, 2586, 2588, 2590, 2592, 2594, 2596, 2598, 2600, 2602, 2604, 2606, 2608, 2610, 2612, 2614, 2616, 2618, 2620, 2622, 2624, 2626, 2628, 2630, 2632, 2634, 2636, 2638, 2640, 2642, 2644, 2646, 2648, 2650, 2652, 2654, 2656, 2658, 2660, 2662, 2664, 2666, 2668, 2670, 2672, 2674, 2676, 2678, 2680, 2682, 2684, 2686, 2688, 2690, 2692, 2694, 2696, 2698, 2700, 2702, 2704, 2706, 2708, 2710, 2712, 2714, 2716, 2718, 2720, 2722, 2724, 2726, 2728, 2730, 2732, 2734, 2736, 2738, 2740, 2742, 2744, 2746, 2748, 2750, 2752, 2754, 2756, 2758, 2760, 2762, 2764, 2766, 2768, 2770, 2772, 2774, 2776, 2778, 2780, 2782, 2784, 2786, 2788, 2790, 2792, 2794, 2796, 2798, 2800, 2802, 2804, 2806, 2808, 2810, 2812, 2814, 2816, 2818, 2820, 2822, 2824, 2826, 2828, 2830, 2832, 2834, 2836, 2838, 2840, 2842, 2844, 2846, 2848, 2850, 2852, 2854, 2856, 2858, 2860, 2862, 2864, 2866, 2868, 2870, 2872, 2874, 2876, 2878, 2880, 2882, 2884, 2886, 2888, 2890, 2892, 2894, 2896, 2898, 2900, 2902, 2904, 2906, 2908, 2910, 2912, 2914, 2916, 2918, 2920, 2922, 2924, 2926, 2928, 2930, 2932, 2934, 2936, 2938, 2940, 2942, 2944, 2946, 2948, 2950, 2952, 2954, 2956, 2958, 2960, 2962, 2964, 2966, 2968, 2970, 2972, 2974, 2976, 2978, 2980, 2982, 2984, 2986, 2988, 2990, 2992, 2994, 2996, 2998, 3000, 3002, 3004, 3006, 3008, 3010, 3012, 3014, 3016, 3018, 3020, 3022, 3024, 3026, 3028, 3030, 3032, 3034, 3036, 3038, 3040, 3042, 3044, 3046, 3048, 3050, 3052, 3054, 3056, 3058, 3060, 3062, 3064, 3066, 3068, 3070, 3072, 3074, 3076, 3078, 3080, 3082, 3084, 3086, 3088, 3090, 3092, 3094, 3096, 3098, 3100, 3102, 3104, 3106, 3108, 3110, 3112, 3114, 3116, 3118, 3120, 3122, 3124, 3126, 3128, 3130, 3132, 3134, 3136, 3138, 3140, 3142, 3144, 3146, 3148, 3150, 3152, 3154, 3156, 3158, 3160, 3162, 3164, 3166, 3168, 3170, 3172, 3174, 3176, 3178, 3180, 3182, 3184, 3186, 3188, 3190, 3192, 3194, 3196, 3198, 3200, 3202, 3204, 3206, 3208, 3210, 3212, 3214, 3216, 3218, 3220, 3222, 3224, 3226, 3228, 3230, 3232, 3234, 3236, 3238, 3240, 3242, 3244, 3246, 3248, 3250, 3252, 3254, 3256, 3258, 3260, 3262, 3264, 3266, 3268, 3270, 3272, 3274, 3276, 3278, 3280, 3282, 3284, 3286, 3288, 3290, 3292, 3294, 3296, 3298, 3300, 3302, 3304, 3306, 3308, 3310, 3312, 3314, 3316, 3318, 3320, 3322, 3324, 3326, 3328, 3330, 3332, 3334, 3336, 3338, 3340, 3342, 3344, 3346, 3348, 3350, 3352, 3354, 3356, 3358, 3360, 3362, 3364, 3366, 3368, 3370, 3372, 3374, 3376, 3378, 3380, 3382, 3384, 3386, 3388, 3390, 3392, 3394, 3396, 3398, 3400, 3402, 3404, 3406, 3408, 3410, 3412, 3414, 3416, 3418, 3420, 3422, 3424, 3426, 3428, 3430, 3432, 3434, 3436, 3438, 3440, 3442, 3444, 3446, 3448, 3450, 3452, 3454, 3456, 3458, 3460, 3462, 3464, 3466, 3468, 3470, 3472, 3474, 3476, 3478, 3480, 3482, 3484, 3486, 3488, 3490, 3492, 3494, 3496, 3498, 3500, 3502, 3504, 3506, 3508, 3510, 3512, 3514, 3516, 3518, 3520, 3522, 3524, 3526, 3528, 3530, 3532, 3534, 3536, 3538, 3540, 3542, 3544, 3546, 3548, 3550, 3552, 3554, 3556, 3558, 3560, 3562, 3564, 3566, 3568, 3570, 3572, 3574, 3576, 3578, 3580, 3582, 3584, 3586, 3588, 3590, 3592, 3594, 3596, 3598, 3600, 3602, 3604, 3606, 3608, 3610, 3612, 3614, 3616, 3618, 3620, 3622, 3624, 3626, 3628, 3630, 3632, 3634, 3636, 3638, 3640, 3642, 3644, 3646, 3648, 3650, 3652, 3654, 3656, 3658, 3660, 3662, 3664, 3666, 3668, 3670, 3672, 3674, 3676, 3678, 3680, 3682, 3684, 3686, 3688, 3690, 3692, 3694, 3696, 3698, 3700, 3702, 3704, 3706, 3708, 3710, 3712, 3714, 3716, 3718, 3720, 3722, 3724, 3726, 3728, 3730, 3732, 3734, 3736, 3738, 3740, 3742, 3744, 3746, 3748, 3750, 3752, 3754, 3756, 3758, 3760, 3762, 3764, 3766, 3768, 3770, 3772, 3774, 3776, 3778, 3780, 3782, 3784, 3786, 3788, 3790, 3792, 3794, 3796, 3798, 3800, 3802, 3804, 3806, 3808, 3810, 3812, 3814, 3816, 3818, 3820, 3822, 3824, 3826, 3828, 3830, 3832, 3834, 3836, 3838, 3840, 3842, 3844, 3846, 3848, 3850, 3852, 3854, 3856, 3858, 3860, 3862, 3864, 3866, 3868, 3870, 3872, 3874, 3876, 3878, 3880, 3882, 3884, 3886, 3888, 3890, 3892, 3894, 3896, 3898, 3900, 3902, 3904, 3906, 3908, 3910, 3912, 3914, 3916, 3918, 3920, 3922, 3924, 3926, 3928, 3930, 3932, 3934, 3936, 3938, 3940, 3942, 3944, 3946, 3948, 3950, 3952, 3954, 3956, 3958, 3960, 3962, 3964, 3966, 3968, 3970, 3972, 3974, 3976, 3978, 3980, 3982, 3984, 3986, 3988, 3990, 3992, 3994, 3996, 3998, 4000, 4002, 4004, 4006, 4008, 4010, 4012, 4014, 4016, 4018, 4020, 4022, 4024, 4026, 4028, 4030, 4032, 4034, 4036, 4038, 4040, 4042, 4044, 4046, 4048, 4050, 4052, 4054, 4056, 4058, 4060, 4062, 4064, 4066, 4068, 4070, 4072, 4074, 4076, 4078, 4080, 4082, 4084, 4086, 4088, 4090, 4092, 4094, 4096, 4098, 4100, 4102, 4104, 4106, 4108, 4110, 4112, 4114, 4116, 4118, 4120, 4122, 4124, 4126, 4128, 4130, 4132, 4134, 4136, 4138, 4140, 4142, 4144, 4146, 4148, 4150, 4152, 4154, 4156, 4158, 4160, 4162, 4164, 4166, 4168, 4170, 4172, 4174, 4176, 4178, 4180, 4182, 4184, 4186, 4188, 4190, 4192, 4194, 4196, 4198, 4200, 4202, 4204, 4206, 4208, 4210, 4212, 4214, 4216, 4218, 4220, 4222, 4224, 4226, 4228, 4230, 4232, 4234, 4236, 4238, 4240, 4242, 4244, 4246, 4248, 4250, 4252, 4254, 4256, 4258, 4260, 4262, 4264, 4266, 4268, 4270, 4272, 4274, 4276, 4278, 4280, 4282, 4284, 4286, 4288, 4290, 4292, 4294, 4296, 4298, 4300, 4302, 4304, 4306, 4308, 4310, 4312, 4314, 4316, 4318, 4320, 4322, 4324, 4326, 4328, 4330, 4332, 4334, 4336, 4338, 4340, 4342, 4344, 4346, 4348, 4350, 4352, 4354, 4356, 4358, 4360, 4362, 4364, 4366, 4368, 4370, 4372, 4374, 4376, 4378, 4380, 4382, 4384, 4386, 4388, 4390, 4392, 4394, 4396, 4398, 4400, 4402, 4404, 4406, 4408, 4410, 4412, 4414, 4416, 4418, 4420, 4422, 4424, 4426, 4428, 4430, 4432, 4434, 4436, 4438, 4440, 4442, 4444, 4446, 4448, 4450, 4452, 4454, 4456, 4458, 4460, 4462, 4464, 4466, 4468, 4470, 4472, 4474, 4476, 4478, 4480, 4482, 4484, 4486, 4488, 4490, 4492, 4494, 4496, 4498, 4500, 4502, 4504, 4506, 4508, 4510, 4512, 4514, 4516, 4518, 4520, 4522, 4524, 4526, 4528, 4530, 4532, 4534, 4536, 4538, 4540, 4542, 4544, 4546, 4548, 4550, 4552, 4554, 4556, 4558, 4560, 4562, 4564, 4566, 4568, 4570, 4572, 4574, 4576, 4578, 4580, 4582, 4584, 4586, 4588, 4590, 4592, 4594, 4596, 4598, 4600, 4602, 4604, 4606, 4608, 4610, 4612, 4614, 4616, 4618, 4620, 4622, 4624, 4626, 4628, 4630, 4632, 4634, 4636, 4638, 4640, 4642, 4644, 4646, 4648, 4650, 4652, 4654, 4656, 4658, 4660, 4662, 4664, 4666, 4668, 4670, 4672, 4674, 4676, 4678, 4680, 4682, 4684, 4686, 4688, 4690, 4692, 4694, 4696, 4698, 4700, 4702, 4704, 4706, 4708, 4710, 4712, 4714, 4716, 4718, 4720; 4722, 4724, 4726, 4728, 4730, 4732, 4734, 4736, 4738, 4740, 4742, 4744, 4746, 4748, 4750, 4752, 4754, 4756, 4758, 4760, 4762, 4764, 4766, 4768, 4770, 4772, 4774, 4776, 4778, 4780, 4782, 4784, 4786, 4788, 4790, 4792, 4794, 4796, 4798, 4800, 4802, 4804, 4806, 4808, 4810, 4812, 4814, 4816, 4818, 4820, 4822, 4824, 4826, 4828, 4830, 4832, 4834, 4836, 4838, 4840, 4842, 4844, 4846, 4848, 4850, 4852, 4854, 4856, 4858, 4860, 4862, 4864, 4866, 4868, 4870, 4872, 4874, 4876, 4878, 4880, 4882, 4884, 4886, 4888, 4890, 4892, 4894, 4896, 4898, 4900 4902, 4904, 4906, 4908, 4910, 4912, 4914, 4916, 4918, 4920, 4922, 4924, 4926, 4928, 4930, 4932, 4934, 4936, 4938, 4940, 4942, 4944, 4946, 4948, 4950, 4952, 4954, 4956, 4958, 4960, 4962, 4964, 4966, 4968, 4970, 4972, 4974, 4976, 4978. 4. An antibody which-binds to a protein according to any one of claims 1 to 3. 5. A nucleic acid molecule which encodes a protein according to any one of claims 1 to 3. 6. A nucleic acid molecule according to claim 5, comprising a nucleotide sequence selected from the group consisting of SEQ IDs 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111,113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581; 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, 679, 681, 683, 685, 687, 689, 691, 693, 695, 697, 699, 701, 703, 705, 707, 709, 711, 713, 715, 717, 719, 721, 723, 725, 727, 729, 731, 733, 735, 737, 739, 741, 743, 745, 747, 749, 751, 753, 755, 757, 759, 761, 763, 765, 767, 769, 771, 773, 775, 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805, 807, 809, 811, 813, 815, 817, 819, 821, 823, 825, 827, 829, 831, 833, 835, 837, 839, 841, 843, 845, 847, 849, 851, 853, 855, 857, 859, 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881, 883, 885, 887, 889, 891, 893, 895, 897, 899, 901, 903, 905, 907, 909, 911, 913, 915, 917, 919, 921, 923, 925, 927, 929, 931, 933, 935, 937, 939, 941, 943, 945, 947, 949, 951, 953, 955, 957, 959, 961, 963, 965, 967, 969, 971, 973, 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 1015, 1017, 1019, 1021, 1023, 1025, 1027, 1029, 1031, 1033, 1035, 1037, 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1063, 1065, 1067, 1069, 1071, 1073, 1075, 1077, 1079, 1081, 1083, 1085, 1087, 1089, 1091, 1093, 1095, 1097, 1099, 1101, 1103, 1105, 1107, 1109, 1111, 1113, 1115, 1117, 1119, 1121, 1123, 1125, 1127, 1129, 1131, 1133, 1135, 1137, 1139, 1141, 1143, 1145, 1147, 1149, 1151, 1153, 1155, 1157, 1159, 1161, 1163, 1165, 1167, 1169, 1171, 1173, 1175, 1177, 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243, 1245, 1247, 1249, 1251, 1253, 1.255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, 1315, 1317, 1319, 1321, 1323, 1325, 1327, 1329, 1331, 1333, 1335, 1337, 1339, 1341, 1343, 1345, 1347, 1349, 1351, 1353, 1355, 1357, 1359, 1361, 1363, 1365, 1367, 1369, 1371, 1373, 1375, 1377, 1379, 1381, 1383, 1385, 1387, 1389, 1391, 1393, 1395, 1397, 1399, 1401, 1403, 1405, 1407, 1409, 1411, 1413, 1415, 1417, 1419, 1421, 1423, 1425, 1427, 1429, 1431, 1433, 1435, 1437, 1439, 1441, 1443, 1445, 1447, 1449, 1451, 1453, 1455, 1457, 1459, 1461, 1463, 1465, 1467, 1469, 1471, 1473, 1475, 1477, 1479, 1481, 1483, 1485, 1487, 1489, 1491, 1493, 1495, 1497, 1499, 1501, 1503, 1505, 1507, 1509, 1511, 1513, 1515, 1517, 1519, 1521, 1523, 1525, 1527, 1529, 1531, 1533, 1535, 1537, 1539, 1541, 1543, 1545, 1547, 1549, 1551, 1553, 1555, 1557, 1559, 1561, 1563, 1565, 1567, 1569, 1571, 1573, 1575, 1577, 1579, 1581, 1583; 1585, 1587, 1589, 1591, 1593, 1595, 1597, 1599, 1601, 1603, 1605, 1607, 1609, 1611, 1613, 1615, 1617, 1619, 1621, 1623, 1625, 1627, 1629, 1631, 1633, 1635, 1637, 1639, 1641, 1643, 1645, 1647, 1649, 1651, 1653, 1655, 1657, 1659, 1661, 1663, 1665, 1667, 1669, 1671, 1673, 1675, 1677, 1679, 1681, 1683, 1685, 1687, 1689, 1691, 1693, 1695, 1697, 1699, 1701, 1703, 1705, 1707, 1709, 1711, 1713, 1715, 1717, 1719, 1721, 1723, 1725, 1727, 1729, 1731, 1733, 1735, 1737, 1739, 1741, 1743, 1745, 1747, 1749, 1751, 1753, 1755, 1757, 1759, 1761, 1763, 1765, 1767, 1769, 1771, 1773, 1775, 1777, 1779, 1781, 1783, 1785, 1787, 1789, 1791, 1793, 1795, 1797, 1799, 1801, 1803, 1805, 1807, 1809, 1811, 1813, 1815, 1817, 1819, 1821, 1823, 1825, 1827, 1829, 1831, 1833, 1835, 1837, 1839, 1841, 1843, 1845, 1847, 1849, 1851, 1853, 1855, 1857, 1859, 1861, 1863, 1865, 1867, 1869, 1871, 1873, 1875, 1877, 1879, 1881, 1883, 1885, 1887, 1889, 1891, 1893, 1895, 1897, 1899, 1901, 1903, 1905, 1907, 1909, 1911, 1913, 1915, 1917, 1919, 1921, 1923, 1925, 1927, 1929, 1931, 1933, 1935, 1937, 1939, 1941, 1943, 1945, 1947, 1949, 1951, 1953, 1955, 1957, 1959, 1961, 1963, 1965, 1967, 1969, 1971, 1973, 1975, 1977, 1979, 1981, 1983, 1985, 1987, 1989, 1991, 1993, 1995, 1997, 1999, 2001, 2003, 2005, 2007, 2009, 2011, 2013, 2015, 2017, 2019, 2021, 2023, 2025, 2027, 2029, 2031, 2033, 2035, 2037, 2039, 2041, 2043, 2045, 2047, 2049, 2051, 2053, 2055, 2057, 2059, 2061, 2063, 2065, 2067, 2069, 2071, 2073, 2075, 2077, 2079, 2081, 2083, 2085, 2087, 2089, 2091, 2093, 2095, 2097, 2099, 2101, 2103, 2105, 2107, 2109, 2111, 2113, 2115, 2117, 2119, 2121, 2123, 2125, 2127, 2129, 2131, 2133, 2135, 2137, 2139, 2141, 2143, 2145, 2147, 2149, 2151, 2153, 2155, 2157, 2159, 2161, 2163, 2165, 2167, 2169, 2171, 2173, 2175, 2177, 2179, 2181, 2183, 2185, 2187, 2189, 2191, 2193, 2195, 2197, 2199, 2201, 2203, 2205, 2207, 2209, 2211, 2213, 2215, 2217, 2219, 2221, 2223, 2225, 2227, 2229, 2231, 2233, 2235, 2237, 2239, 2241, 2243, 2245, 2247, 2249, 2251, 2253, 2255, 2257, 2259, 2261, 2263, 2265, 2267, 2269, 2271, 2273, 2275, 2277, 2279, 2281, 2283, 2285, 2287, 2289, 2291, 2293, 2295, 2297, 2299, 2301, 2303, 2305, 2307, 2309, 2311, 2313, 2315, 2317, 2319, 2321, 2323, 2325, 2327, 2329, 2331, 2333, 2335, 2337, 2339, 2341, 2343, 2345, 2347, 2349, 2351, 2353, 2355, 2357, 2359, 2361, 2363, 2365, 2367, 2369, 2371, 2373, 2375, 2377, 2379, 2381, 2383, 2385, 2387, 2389, 2391, 2393, 2395, 2397, 2399, 2401, 2403, 2405, 2407, 2409, 2411, 2413; 2415, 2417, 2419, 2421, 2423, 2425, 2427, 2429, 2431, 2433, 2435, 2437, 2439, 2441, 2443, 2445, 2447, 2449, 2451, 2453, 2455, 2457, 2459, 2461, 2463, 2465, 2467, 2469, 2471, 2473, 2475, 2477, 2479, 2481, 2483, 2485, 2487, 2489, 2491, 2493, 2495, 2497, 2499, 2501, 2503, 2505, 2507, 2509, 2511, 2513, 2515, 2517, 2519, 2521, 2523, 2525, 2527, 2529, 2531, 2533, 2535, 2537, 2539, 2541, 2543, 2545, 2547, 2549, 2551, 2553, 2555, 2557, 2559, 2561, 2563, 2565, 2567, 2569, 2571, 2573, 2575, 2577, 2579, 2581, 2583, 2585, 2587, 2589, 2591, 2593, 2595, 2597, 2599, 2601, 2603, 2605, 2607, 2609, 2611, 2613, 2615, 2617, 2619, 2621, 2623, 2625, 2627, 2629, 2631, 2633, 2635, 2637, 2639, 2641, 2643, 2645, 2647, 2649, 2651, 2653, 2655, 2657, 2659, 2661, 2663, 2665, 2667, 2669, 2671, 2673, 2675, 2677, 2679, 2681, 2683, 2685, 2687, 2689, 2691, 2693, 2695, 2697, 2699, 2701, 2703, 2705, 2707, 2709, 2711, 2713, 2715, 2717, 2719, 2721, 2723, 2725, 2727, 2729, 2731, 2733, 2735, 2737, 2739, 2741, 2743, 2745, 2747, 2749, 2751, 2753, 2755, 2757, 2759, 2761, 2763, 2765, 2767, 2769, 2771, 2773, 2775, 2777, 2779, 2781, 2783, 2785, 2787, 2789, 2791, 2793, 2795, 2797, 2799, 2801, 2803, 2805, 2807, 2809, 2811, 2813, 2815, 2817, 2819, 2821, 2823, 2825, 2827, 2829, 2831, 2833, 2835, 2837, 2839, 2841, 2843, 2845, 2847, 2849, 2851, 2853, 2855, 2857, 2859, 2861, 2863, 2865, 2867, 2869, 2871, 2873, 2875, 2877, 2879, 2881, 2883, 2885, 2887, 2889, 2891, 2893, 2895, 2897, 2899, 2901, 2903, 2905, 2907, 2909, 2911, 2913, 2915, 2917, 2919, 2921, 2923, 2925, 2927, 2929, 2931, 2933, 2935, 2937, 2939, 2941, 2943, 2945, 2947, 2949, 2951, 2953, 2955, 2957, 2959, 2961, 2963, 2965, 2967, 2969, 2971, 2973, 2975, 2977, 2979, 2981, 2983, 2985, 2987, 2989, 2991, 2993, 2995, 2997, 2999, 3001, 3003, 3005, 3007, 3009, 3011, 3013, 3015, 3017, 3019, 3021, 3023, 3025, 3027, 3029, 3031, 3033, 3035, 3037, 3039, 3041, 3043, 3045, 3047, 3049, 3051, 3053, 3055, 3057, 3059, 3061, 3063, 3065, 3067, 3069, 3071, 3073, 3075, 3077, 3079, 3081, 3083, 3085, 3087, 3089, 3091, 3093, 3095, 3097, 3099, 3101, 3103, 3105, 3107, 3109, 3111, 3113, 3115, 3117, 3119, 3121, 3123, 3125, 3127, 3129, 3131, 3133, 3135, 3137, 3139, 3141, 3143, 3145, 3147, 3149, 3151, 3153, 3155, 3157, 3159, 3161, 3163, 3165, 3167, 3169, 3171, 3173, 3175, 3177, 3179, 3181, 3183, 3185, 3187, 3189, 3191, 3193, 3195, 3197, 3199, 3201, 3203, 3205, 3207, 3209, 3211, 3213, 3215, 3217, 3219, 3221, 3223, 3225, 3227, 3229, 3231, 3233, 3235, 3237, 3239, 3241, 3243, 3245, 3247, 3249, 3251, 3253, 3255, 3257, 3259, 3261, 3263, 3265, 3267, 3269, 3271, 3273, 3275, 3277, 3279, 3281, 3283, 3285, 3287, 3289, 3291, 3293, 3295, 3297, 3299, 3301, 3303, 3305, 3307, 3309, 3311, 3313, 3315, 3317, 3319, 3321, 3323, 3325, 3327, 3329, 3331, 3333, 3335, 3337, 3339, 3341, 3343, 3345, 3347, 3349, 3351, 3353, 3355, 3357, 3359, 3361, 3363, 3365, 3367, 3369, 3371, 3373, 3375, 3377, 3379, 3381, 3383, 3385, 3387, 3389, 3391, 3393, 3395, 3397, 3399, 3401, 3403, 3405, 3407, 3409, 3411, 3413, 3415, 3417, 3419, 3421, 3423, 3425, 3427, 3429, 3431, 3433, 3435, 3437, 3439, 3441, 3443, 3445, 3447, 3449, 3451, 3453, 3455, 3457, 3459, 3461, 3463, 3465, 3467, 3469, 3471, 3473, 3475, 3477, 3479, 3481, 3483, 3485, 3487, 3489, 3491, 3493, 3495, 3497, 3499, 3501, 3503, 3505, 3507, 3509, 3511, 3513, 3515, 3517, 3519, 3521, 3523, 3525, 3527, 3529, 3531, 3533, 3535, 3537, 3539, 3541, 3543, 3545, 3547, 3549, 3551, 3553, 3555, 3557, 3559, 3561, 3563, 3565, 3567, 3569, 3571, 3573, 3575, 3577, 3579, 3581, 3583, 3585, 3587, 3589, 3591, 3593, 3595, 3597, 3599, 3601, 3603, 3605, 3607, 3609, 3611, 3613, 3615, 3617, 3619, 3621, 3623, 3625, 3627, 3629, 3631, 3633, 3635, 3637, 3639, 3641, 3643, 3645, 3647, 3649, 3651, 3653, 3655, 3657, 3659, 3661, 3663, 3665, 3667, 3669, 3671, 3673, 3675, 3677, 3679, 3681, 3683, 3685, 3687, 3689, 3691, 3693, 3695, 3697, 3699, 3701, 3703, 3705, 3707, 3709, 3711, 3713, 3715, 3717, 3719, 3721, 3723, 3725, 3727, 3729, 3731, 3733, 3735, 3737, 3739, 3741, 3743, 3745, 3747, 3749, 3751, 3753, 3755, 3757, 3759, 3761, 3763, 3765, 3767, 3769, 3771, 3773, 3775, 3777, 3779, 3781, 3783, 3785, 3787, 3789, 3791, 3793, 3795, 3797, 3799, 3801, 3803, 3805, 3807, 3809, 3811, 3813, 3815, 3817, 3819, 3821, 3823, 3825, 3827, 3829, 3831, 3833, 3835, 3837, 3839, 3841, 3843, 3845, 3847, 3849, 3851, 3853, 3855, 3857, 3859, 3861, 3863, 3865, 3867, 3869, 3871, 3873, 3875, 3877, 3879, 3881, 3883, 3885, 3887, 3889, 3891, 3893, 3895, 3897, 3899, 3901, 3903, 3905, 3907, 3909, 3911, 3913, 3915, 3917, 3919, 3921, 3923, 3925, 3927, 3929, 3931, 3933, 3935, 3937, 3939, 3941, 3943, 3945, 3947, 3949, 3951, 3953, 3955, 3957, 3959, 3961, 3963, 3965, 3967, 3969, 3971, 3973, 3975, 3977, 3979, 3981, 3983, 3985, 3987, 3989, 3991, 3993, 3995, 3997, 3999, 4001, 4003, 4005, 4007, 4009, 4011, 4013, 4015, 4017, 4019, 4021, 4023, 4025, 4027, 4029, 4031, 4033, 4035, 4037, 4039, 4041, 4043, 4045, 4047, 4049, 4051, 4053, 4055, 4057, 4059, 4061, 4063, 4065, 4067, 4069, 4071, 4073, 4075, 4077, 4079, 4081, 4083, 4085, 4087, 4089, 4091, 4093, 4095, 4097, 4099, 4101, 4103, 4105, 4107, 4109, 4111, 4113, 4115, 4117, 4119, 4121, 4123, 4125, 4127, 4129, 4131, 4133, 4135, 4137, 4139, 4141, 4143, 4145, 4147, 4149, 4151, 4153, 4155, 4157, 4159, 4161, 4163, 4165, 4167, 4169, 4171, 4173, 4175, 4177, 4179, 4181, 4183, 4185, 4187, 4189, 4191, 4193, 4195, 4197, 4199, 4201, 4203, 4205, 4207, 4209, 4211, 4213, 4215, 4217, 4219, 4221, 4223, 4225, 4227, 4229, 4231, 4233, 4235, 4237, 4239, 4241, 4243, 4245, 4247, 4249, 4251, 4253, 4255, 4257, 4259, 4261, 4263, 4265, 4267, 4269, 4271, 4273, 4275, 4277, 4279, 4281, 4283, 4285, 4287, 4289, 4291, 4293, 4295, 4297, 4299, 4301, 4303, 4305, 4307, 4309, 4311, 4313, 4315, 4317, 4319, 4321, 4323, 4325, 4327, 4329, 4331, 4333, 4335, 4337, 4339, 4341, 4343, 4345, 4347, 4349, 4351, 4353, 4355, 4357, 4359, 4361, 4363, 4365, 4367, 4369, 4371, 4373, 4375, 4377, 4379, 4381, 4383, 4385, 4387, 4389, 4391, 4393, 4395, 4397, 4399, 4401, 4403, 4405, 4407, 4409, 4411, 4413, 4415, 4417, 4419, 4421, 4423, 4425, 4427, 4429, 4431, 4433, 4435, 4437, 4439, 4441, 4443, 4445, 4447, 4449, 4451, 4453, 4455, 4457, 4459, 4461, 4463, 4465, 4467, 4469, 4471, 4473, 4475, 4477, 4479, 4481, 4483, 4485, 4487, 4489, 4491, 4493, 4495, 4497, 4499, 4501, 4503, 4505, 4507, 4509, 4511, 4513, 4515, 4517, 4519, 4521, 4523, 4525, 4527, 4529, 4531, 4533, 4535, 4537, 4539, 4541, 4543, 4545, 4547, 4549, 4551, 4553, 4555, 4557, 4559, 4561, 4563, 4565, 4567, 4569, 4571, 4573, 4575, 4577, 4579, 4581, 4583, 4585, 4587, 4589, 4591, 4593, 4595, 4597, 4599, 4601, 4603, 4605, 4607, 4609, 4611, 4613, 4615, 4617, 4619, 4621, 4623, 4625, 4627, 4629, 4631, 4633, 4635, 4637, 4639, 4641, 4643, 4645, 4647, 4649, 4651, 4653, 4655, 4657, 4659, 4661, 4663, 4665, 4667, 4669, 4671, 4673, 4675, 4677, 4679, 4681, 4683, 4685, 4687, 4689, 4691, 4693, 4695, 4697, 4699, 4701, 4703, 4705, 4707, 4709, 4711, 4713, 4715, 4717, 4719, 4721, 4723, 4725, 4727, 4729, 4731, 4733, 4735, 4737, 4739, 4741, 4743, 4745, 4747, 4749, 4751, 4753, 4755, 4757, 4759, 4761, 4763, 4765, 4767, 4769, 4771, 4773, 4775, 4777, 4779, 4781, 4783, 4785, 4787, 4789, 4791, 4793, 4795, 4797, 4799, 4801, 4803, 4805, 4807, 4809, 4811, 4813, 4815, 4817, 4819, 4821, 4823, 4825, 4827, 4829, 4831, 4833, 4835, 4837, 4839, 4841, 4843, 4845, 4847, 4849, 4851, 4853, 4855, 4857, 4859, 4861, 4863, 4865, 4867, 4869, 4871, 4873, 4875, 4877, 4879, 4881, 4883, 4885, 4887, 4889, 4891; 4893, 4895, 4897, 4899, 4901, 4903, 4905, 4907, 4909, 4911, 4913, 4915, 4917, 4919, 4921, 4923, 4925, 4927, 4929, 4931, 4933, 4935, 4937, 4939, 4941, 4943, 4945, 4947, 4949, 4951, 4953, 4955, 4957, 4959, 4961, 4963, 4965, 4967, 4969, 4971, 4973, 4975, 4977. 7. A nucleic acid molecule comprising a fragment of a nucleotide sequence selected from the group consisting of SEQ IDs 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 24.9, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, 679, 681, 683, 685, 687, 689, 691, 693, 695, 697, 699, 701, 703, 705, 707, 709, 711, 713, 715, 717, 719, 721, 723, 725, 727, 729, 731, 733, 735, 737, 739, 741, 743, 745, 747, 749, 751, 753, 755, 757, 759, 761, 763, 765, 767, 769, 771, 773, 775, 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805, 807, 809, 811, 813, 815, 817, 819, 821, 823, 825, 827, 829, 831, 833, 835, 837, 839, 841, 843, 845, 847, 849, 851, 853, 855, 857, 859, 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881, 883, 885, 887, 889, 891, 893, 895, 897, 899, 901, 903, 905, 907, 909, 911, 913, 915, 917, 919, 921, 923, 925, 927, 929, 931, 933, 935, 937, 939, 941, 943, 945, 947, 949, 951, 953, 955, 957, 959, 961, 963, 965, 967, 969, 971, 973, 975, 977, 979, 981, 983, 985, 987, 989, 991, 993, 995, 997, 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 1015, 1017, 1019, 1021, 1023, 1025, 1027, 1029, 1031, 1033, 1035, 1037, 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1063, 1065, 1067, 1069, 1071, 1073, 1075, 1077, 1079, 1081, 1083, 1085, 1087, 1089, 1091, 1093, 1095, 1097, 1099, 1101, 1103, 1105, 1107, 1109, 1111, 1113, 1115, 1117, 1119, 1121, 1123, 1125, 1127, 1129, 1131, 1133, 1135, 1137, 1139, 1141, 1143, 1145, 1147, 1149, 1151, 1153, 1155, 1157, 1159, 1161, 1163, 1165, 1167, 1169, 1171, 1173, 1175, 1177, 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243, 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, 1315, 1317, 1319, 1321, 1323, 1325, 1327, 1329, 1331, 1333, 1335, 1337, 1339, 1341, 1343, 1345, 1347, 1349, 1351, 1353, 1355, 1357, 1359, 1361, 1363, 1365, 1367, 1369, 1371, 1373, 1375, 1377, 1379, 1381, 1383, 1385, 1387, 1389, 1391, 1393, 1395, 1397, 1399, 1401, 1403, 1405, 1407, 1409, 1411, 1413, 1415, 1417, 1419, 1421, 1423, 1425, 1427, 1429, 1431, 1433, 1435, 1437, 1439, 1441, 1443, 1445, 1447, 1449, 1451, 1453, 1455, 1457, 1459, 1461, 1463, 1465, 1467, 1469, 1471, 1473, 1475, 1477, 1479, 1481, 1483, 1485, 1487, 1489, 1491, 1493, 1495, 1497, 1499, 1501, 1503, 1505, 1507, 1509, 1511, 1513, 1515, 1517, 1519, 1521, 1523, 1525, 1527, 1529, 1531, 1533, 1535, 1537, 1539, 1541, 1543, 1545, 1547, 1549, 1551, 1553, 1555, 1557, 1559, 1561, 1563, 1565, 1567, 1569, 1571, 1573, 1575, 1577, 1579, 1581, 1583, 1585, 1587, 1589, 1591, 1593, 1595, 1597, 1599, 1601, 1603, 1605, 1607, 1609, 1611, 1613, 1615, 1617, 1619, 1621, 1623, 1625, 1627, 1629, 1631, 1633, 1635, 1637, 1639, 1641, 1643, 1645, 1647, 1649, 1651, 1653, 1655, 1657, 1659, 1661, 1663, 1665, 1667, 1669, 1671, 1673, 1675, 1677, 1679, 1681, 1683, 1685, 1687, 1689, 1691, 1693, 1695, 1697, 1699, 1701, 1703, 1705, 1707, 1709, 1711, 1713, 1715, 1717, 1719, 1721, 1723, 1725, 1727, 1729, 1731, 1733, 1735, 1737, 1739, 1741, 1743, 1745, 1747, 1749, 1751, 1753, 1755, 1757, 1759, 1761, 1763, 1765, 1767, 1769, 1771, 1773, 1775, 1777, 1779, 1781, 1783, 1785, 1787, 1789, 1791, 1793, 1795, 1797, 1799, 1801, 1803, 1805, 1807, 1809, 1811, 1813, 1815, 1817, 1819, 1821, 1823, 1825, 1827, 1829, 1831, 1833, 1835, 1837, 1839, 1841, 1843, 1845, 1847, 1849, 1851, 1853, 1855, 1857, 1859, 1861, 1863, 1865, 1867, 1869, 1871, 1873, 1875, 1877, 1879, 1881, 1883, 1885, 1887, 1889, 1891, 1893, 1895, 1897, 1899, 1901, 1903, 1905, 1907, 1909, 1911, 1913, 1915, 1917, 1919, 1921, 1923, 1925, 1927, 1929, 1931, 1933, 1935, 1937, 1939, 1941, 1943, 1945, 1947, 1949, 1951, 1953, 1955, 1957, 1959, 1961, 1963, 1965, 1967, 1969, 1971, 1973, 1975, 1977, 1979, 1981, 1983, 1985, 1987, 1989, 1991, 1993, 1995, 1997, 1999, 2001, 2003, 2005, 2007, 2009, 2011, 2013, 2015, 2017, 2019, 2021, 2023, 2025, 2027, 2029, 2031, 2033, 2035, 2037, 2039, 2041, 2043, 2045, 2047, 2049, 2051, 2053, 2055, 2057, 2059, 2061, 2063, 2065, 2067, 2069, 2071, 2073, 2075, 2077, 2079, 2081, 2083, 2085, 2087, 2089, 2091, 2093, 2095, 2097, 2099, 2101, 2103, 2105, 2107, 2109, 2111, 2113, 2115, 2117, 2119, 2121, 2123, 2125, 2127, 2129, 2131, 2133, 2135, 2137, 2139, 2141, 2143, 2145, 2147, 2149, 2151, 2153, 2155, 2157, 2159, 2161, 2163, 2165, 2167, 2169, 2171, 2173, 2175, 2177, 2179, 2181, 2183, 2185, 2187, 2189, 2191, 2193, 2195, 2197, 2199, 2201, 2203, 2205, 2207, 2209, 2211, 2213, 2215, 2217, 2219, 2221, 2223, 2225, 2227, 2229, 2231, 2233, 2235, 2237, 2239, 2241, 2243, 2245, 2247, 2249, 2251, 2253, 2255, 2257, 2259, 2261, 2263, 2265, 2267, 2269, 2271, 2273, 2275, 2277, 2279, 2281, 2283, 2285, 2287, 2289, 2291, 2293, 2295, 2297, 2299, 2301, 2303, 2305, 2307, 2309, 2311, 2313, 2315, 2317, 2319, 2321, 2323, 2325, 2327, 2329, 2331, 2333, 2335, 2337, 2339, 2341, 2343, 2345, 2347, 2349, 2351, 2353, 2355, 2357, 2359, 2361, 2363, 2365, 2367, 2369, 2371, 2373, 2375, 2377, 2379, 2381, 2383, 2385, 2387, 2389, 2391, 2393, 2395, 2397, 2399, 2401, 2403, 2405, 2407, 2409, 2411, 2413, 2415, 2417, 2419, 2421, 2423, 2425, 2427, 2429, 2431, 2433, 2435, 2437, 2439, 2441, 2443, 2445, 2447, 2449, 2451, 2453, 2455, 2457, 2459, 2461, 2463, 2465, 2467, 2469, 2471, 2473, 2475, 2477, 2479, 2481, 2483, 2485, 2487, 2489, 2491, 2493, 2495, 2497, 2499, 2501, 2503, 2505, 2507, 2509, 2511, 2513, 2515, 2517, 2519, 2521, 2523, 2525, 2527, 2529, 2531, 2533, 2535, 2537, 2539, 2541, 2543, 2545, 2547, 2549, 2551, 2553, 2555, 2557, 2559, 2561, 2563, 2565, 2567, 2569, 2571, 2573, 2575, 2577, 2579, 2581, 2583, 2585, 2587, 2589, 2591, 2593, 2595, 2597, 2599, 2601, 2603, 2605, 2607, 2609, 2611, 2613, 2615, 2617, 2619, 2621, 2623, 2625, 2627, 2629, 2631, 2633, 2635, 2637, 2639, 2641, 2643, 2645, 2647, 2649, 2651, 2653, 2655, 2657, 2659, 2661, 2663, 2665, 2667, 2669, 2671, 2673, 2675, 2677, 2679, 2681, 2683, 2685, 2687, 2689, 2691, 2693, 2695, 2697, 2699, 2701, 2703, 2705, 2707, 2709, 2711, 2713, 2715, 2717, 2719, 2721, 2723, 2725, 2727, 2729, 2731, 2733, 2735, 2737, 2739, 2741, 2743, 2745, 2747, 2749, 2751, 2753, 2755, 2757, 2759, 2761, 2763, 2765, 2767, 2769, 2771, 2773, 2775, 2777, 2779, 2781, 2783, 2785, 2787, 2789, 2791, 2793, 2795, 2797, 2799, 2801, 2803, 2805, 2807, 2809, 2811, 2813, 2815, 2817, 2819, 2821, 2823, 2825, 2827, 2829, 2831, 2833, 2835, 2837, 2839, 2841, 2843, 2845, 2847, 2849, 2851, 2853, 2855, 2857, 2859, 2861, 2863, 2865, 2867, 2869, 2871, 2873, 2875, 2877, 2879, 2881, 2883, 2885, 2887, 2889, 2891, 2893, 2895, 2897, 2899, 2901, 2903, 2905, 2907, 2909, 2911, 2913, 2915, 2917, 2919, 2921, 2923, 2925, 2927, 2929, 2931, 2933, 2935, 2937, 2939, 2941, 2943, 2945, 2947, 2949, 2951, 2953, 2955, 2957, 2959, 2961, 2963, 2965, 2967, 2969, 2971, 2973, 2975, 2977, 2979, 2981, 2983, 2985, 2987, 2989, 2991, 2993, 2995, 2997, 2999, 3001, 3003, 3005, 3007, 3009, 3011, 3013, 3015, 3017, 3019, 3021, 3023, 3025, 3027, 3029, 3031, 3033, 3035, 3037, 3039, 3041, 3043, 3045, 3047, 3049, 3051, 3053, 3055, 3057, 3059, 3061, 3063, 3065, 3067, 3069, 3071, 3073, 3075, 3077, 3079, 3081, 3083, 3085, 3087, 3089, 3091, 3093, 3095, 3097, 3099, 3101, 3103, 3105, 3107, 3109, 3111, 3113, 3115, 3117, 3119, 3121, 3123, 3125, 3127, 3129, 3131, 3133, 3135, 3137, 3139, 3141, 3143, 3145, 3147, 3149, 3151, 3153, 3155, 3157, 3159, 3161, 3163, 3165, 3167, 3169, 3171, 3173, 3175, 3177, 3179, 3181, 3183, 3185, 3187, 3189, 3191, 3193, 3195, 3197, 3199, 3201, 3203, 3205, 3207, 3209, 3211, 3213, 3215, 3217, 3219, 3221, 3223, 3225, 3227, 3229, 3231, 3233, 3235, 3237, 3239, 3241, 3243, 3245, 3247, 3249, 3251, 3253, 3255, 3257, 3259, 3261, 3263, 3265, 3267, 3269, 3271, 3273, 3275, 3277, 3279, 3281, 3283, 3285, 3287, 3289, 3291, 3293, 3295, 3297, 3299, 3301, 3303, 3305, 3307, 3309, 3311, 3313, 3315, 3317, 3319, 3321, 3323, 3325, 3327, 3329, 3331, 3333, 3335, 3337, 3339, 3341, 3343, 3345, 3347, 3349, 3351, 3353, 3355, 3357, 3359, 3361, 3363, 3365, 3367, 3369, 3371, 3373, 3375, 3377, 3379, 3381, 3383, 3385, 3387, 3389, 3391, 3393, 3395, 3397, 3399, 3401, 3403, 3405, 3407, 3409, 3411, 3413, 3415, 3417, 3419, 3421, 3423, 3425, 3427, 3429, 3431, 3433, 3435, 3437, 3439, 3441, 3443, 3445, 3447, 3449, 3451, 3453, 3455, 3457, 3459, 3461, 3463, 3465, 3467, 3469, 3471, 3473, 3475, 3477, 3479, 3481, 3483, 3485, 3487, 3489, 3491, 3493, 3495, 3497, 3499, 3501, 3503, 3505, 3507, 3509, 3511, 3513, 3515, 3517, 3519, 3521, 3523, 3525, 3527, 3529, 3531, 3533, 3535, 3537, 3539, 3541, 3543, 3545, 3547, 3549, 3551, 3553, 3555, 3557, 3559, 3561, 3563, 3565, 3567, 3569, 3571, 3573, 3575, 3577, 3579, 3581, 3583, 3585, 3587, 3589, 3591, 3593, 3595, 3597, 3599, 3601, 3603, 3605, 3607, 3609, 3611, 3613, 3615, 3617, 3619, 3621, 3623, 3625, 3627, 3629, 3631, 3633, 3635, 3637, 3639, 3641, 3643, 3645, 3647, 3649, 3651, 3653, 3655, 3657, 3659, 3661, 3663, 3665, 3667, 3669, 3671, 3673, 3675, 3677, 3679, 3681, 3683, 3685, 3687, 3689, 3691, 3693, 3695, 3697, 3699, 3701, 3703, 3705, 3707, 3709, 3711, 3713, 3715, 3717, 3719, 3721, 3723, 3725, 3727, 3729, 3731, 3733, 3735, 3737, 3739, 3741, 3743, 3745, 3747, 3749, 3751, 3753, 3755, 3757, 3759, 3761, 3763, 3765, 3767, 3769, 3771, 3773, 3775, 3777, 3779, 3781, 3783, 3785, 3787, 3789, 3791, 3793, 3795, 3797, 3799, 3801, 3803, 3805, 3807, 3809, 3811, 3813, 3815, 3817, 3819, 3821, 3823, 3825, 3827, 3829, 3831, 3833, 3835, 3837, 3839, 3841, 3843, 3845, 3847, 3849, 3851, 3853, 3855, 3857, 3859, 3861, 3863, 3865, 3867, 3869, 3871, 3873, 3875, 3877, 3879, 3881, 3883, 3885, 3887, 3889, 3891, 3893, 3895, 3897, 3899, 3901, 3903, 3905, 3907, 3909, 3911, 3913, 3915, 3917, 3919, 3921, 3923, 3925, 3927, 3929, 3931, 3933, 3935, 3937, 3939, 3941, 3943, 3945, 3947, 3949, 3951, 3953, 3955, 3957, 3959, 3961, 3963, 3965, 3967, 3969, 3971, 3973, 3975, 3977, 3979, 3981, 3983, 3985, 3987, 3989, 3991, 3993, 3995, 3997, 3999, 4001, 4003, 4005, 4007, 4009, 4011, 4013, 4015, 4017, 4019, 4021, 4023, 4025, 4027, 4029, 4031, 4033, 4035, 4037, 4039, 4041, 4043, 4045, 4047, 4049, 4051, 4053, 4055, 4057, 4059, 4061, 4063, 4065, 4067, 4069, 4071, 4073, 4075, 4077, 4079, 4081, 4083, 4085, 4087, 4089, 4091, 4093, 4095, 4097, 4099, 4101, 4103, 4105, 4107, 4109, 4111, 4113, 4115, 4117, 4119, 4121, 4123, 4125, 4127, 4129, 4131, 4133, 4135, 4137, 4139, 4141, 4143, 4145, 4147, 4149, 4151, 4153, 4155, 4157, 4159, 4161, 4163, 4165, 4167, 4169, 4171, 4173, 4175, 4177, 4179, 4181, 4183, 4185, 4187, 4189, 4191, 4193, 4195, 4197, 4199, 4201, 4203, 4205, 4207, 4209, 4211, 4213, 4215, 4217, 4219, 4221, 4223, 4225, 4227, 4229, 4231, 4233, 4235, 4237, 4239, 4241, 4243, 4245, 4247, 4249, 4251, 4253, 4255, 4257, 4259, 4261, 4263, 4265, 4267, 4269, 4271, 4273, 4275, 4277, 4279, 4281, 4283, 4285, 4287, 4289, 4291, 4293, 4295, 4297, 4299, 4301, 4303, 4305, 4307, 4309, 4311, 4313, 4315, 4317, 4319, 4321, 4323, 4325, 4327, 4329, 4331, 4333, 4335, 4337, 4339, 4341, 4343, 4345, 4347, 4349, 4351, 4353, 4355, 4357, 4559, 4361, 4363, 4365, 4367, 4369, 4371, 4373, 4375, 4377, 4379, 4381, 4383, 4385, 4387, 4389, 4391, 4393, 4395, 4397, 4399, 4401, 4403, 4405, 4407, 4409, 4411, 4413, 4415, 4417, 4419, 4421, 4423, 4425, 4427, 4429, 4431, 4433, 4435, 4437, 4439, 4441, 4443, 4445, 4447, 4449, 4451, 4453, 4455, 4457, 4459, 4461, 4463, 4465, 4467, 4469, 4471, 4473, 4475, 4477, 4479, 4481, 4483, 4485, 4487, 4489, 4491, 4493, 4495, 4497, 4499, 4501, 4503, 4505, 4507, 4509, 4511, 4513, 4515, 4517, 4519, 4521, 4523, 4525, 4527, 4529, 4531, 4533, 4535, 4537, 4539, 4541, 4543, 4545, 4547, 4549, 4551, 4553, 4555, 4557, 4559, 4561, 4563, 4565, 4567, 4569, 4571, 4573, 4575, 4577, 4579, 4581, 4583, 4585, 4587, 4589, 4591, 4593, 4595, 4597, 4599, 4601, 4603, 4605, 4607, 4609, 4611, 4613, 4615, 4617, 4619, 4621, 4623, 4625, 4627, 4629, 4631, 4633, 4635, 4637, 4639, 4641, 4643, 4645, 4647, 4649, 4651, 4653, 4655, 4657, 4659, 4661, 4663, 4665, 4667, 4669, 4671, 4673, 4675, 4677, 4679, 4681, 4683, 4685, 4687, 4689, 4691, 4693, 4695, 4697, 4699, 4701, 4703, 4705, 4707, 4709, 4711, 4713, 4715, 4717, 4719, 4721, 4723, 4725, 4727, 4729, 4731, 4733, 4735, 4737, 4739, 4741, 4743, 4745, 4747, 4749, 4751, 4753, 4755, 4757, 4759, 4761, 4763, 4765; 4767, 4769, 4771, 4773, 4775, 4777, 4779, 4781, 4783, 4785, 4787, 4789, 4791, 4793, 4795, 4797, 4799, 4801, 4803, 4805, 4807, 4809, 4811, 4813, 4815, 4817, 4819, 4821, 4823, 4825, 4827, 4829, 4831, 4833, 4835, 4837, 4839, 4841, 4843, 4845, 4847, 4849, 4851, 4853, 4855, 4857, 4859, 4861, 4863, 4865, 4867, 4869, 4871, 4873, 4875, 4877, 4879, 4881, 4883, 4885, 4887, 4889, 4891, 4893, 4895, 4897, 4899, 4901, 4903, 4905, 4907, 4909, 4911, 4913, 4915, 4917, 4919, 4921, 4923, 4925, 4927, 4929, 4931, 4933, 4935, 4937, 4939, 4941, 4943, 4945, 4947, 4949, 4951, 4953, 4955, 4957, 4959, 4961, 4963, 4965, 4967, 4969, 4971, 4973, 4975, 4977. 8. A nucleic acid molecule comprising a nucleotide sequence complementary to a nucleic acid molecule according to any one of claims 5 to 7. 9. A nucleic acid molecule comprising a nucleotide sequences having 50% or greater sequence identity to a nucleic acid molecule according to any one of claims 5 to 8. 10. A nucleic acid molecule which can hybridise to a nucleic acid molecule according to any one of claims 5 to 9 under high stringency conditions. 11. A composition comprising a protein, a nucleic acid molecule, or an antibody according to any preceding claim. 12. A composition according to claim 11 being a vaccine composition or a diagnostic composition. 13. A composition according to claim 11 or claim 12 for use as a pharmaceutical. 14. The use of a composition according to claim 13 in the manufacture of a medicament for the treatment or prevention of a disease or infection due to streptococcus bacteria, particularly S.pneumoniae. 15. The use of claim 14, wherein the disease is meningitis, pneumonia, sepsis, otitis media or an ear infection. 16. A method of treating a patient, comprising administering to the patient a therapeutically effective amount of a composition according to claim 13. 17. A kit comprising primers for amplifying a target sequence contained within a Streptococcus nucleic acid sequence, the kit comprising a first primer and a second primer, wherein the first primer is substantially complementary to said target sequence and the second primer is substantially complementary to a complement of said target sequence, wherein the parts of said primers which have substantial complementarity define the termini of the target sequence to be amplified, and wherein the first and/or second primer is a nucleic acid according to any one of claims 5 to 10, or a fragment of between 8 and 100 nucleotides of SEQ ID 4979. 18. A kit comprising first and second single-stranded oligonucleotides which allow amplification of a Streptococcus template nucleic acid sequence contained in a single- or double-stranded nucleic acid (or mixture thereof), wherein: (a) the first oligonucleotide comprises a primer sequence which is substantially complementary to said template nucleic acid sequence; (b) the second oligonucleotide comprises a primer sequence which is substantially complementary to the complement of said template nucleic acid sequence; (c) the first oligonucleotide and/or the second oligonucleotide comprise(s) sequence which is not complementary to said template nucleic acid; (d) said primer sequences define the termini of the template sequence to be amplified; and (e) the first and/or second oligonucleotide is a nucleic acid according to any one of claims 5 to 10, or a fragment of between 8 and 100 nucleotides of SEQ ID 4979. 19. A hybrid protein represented by the formula NH2-A-[-X-L-]a, —B—COOH, wherein X is the amino acid sequence of a protein according to claim 1, claim 2 or claim 3, L is an optional linker amino acid sequence, A is an optional N-terminal amino acid sequence, B is an optional C-terminal amino acid sequence, and n is an integer greater than 1. 20. An assay comprising the steps of contacting a test compound with a protein according to any one of claims 1 to 3, and determining whether the test compound binds to said protein. 21. The composition of claim 13 further comprising one or more of the following antigens: a protein antigen from Helicobacter pylori; a protein antigen from N.meningitidis serogroup B; an outer-membrane vesicle (OMV) preparation from N.meningitidis; a saccharide antigen from N.meningitidis serogroup A, C, W 135 and/or Y; a saccharide antigen from Streptococcus pneumoniae; an antigen from hepatitis A virus; an antigen from hepatitis B virus; an antigen from hepatitis C virus; an antigen from Bordetelia pertussis; a diphtheria antigen; a tetanus antigen; a saccharide antigen from Haemophilus influenzae B. an antigen from N.gonorrhoeae; an antigen from Chlamydia pneumoniae; an antigen from Streptococcus agalactiae; an antigen from Streptococcus pyogenes an antigen from Chlamydia trachonzatis an antigen from Porphyromonas gingivalis; polio antigen(s); rabies antigen(s); measles, mumps and/or rubella antigens; influenza antigen(s); an antigen from Moraxella catarrhalis; and/or an antigen from Staphylococcus aureus. 22. A composition comprising two or more proteins of any one of claims 1 to 3. 23. A S.pneumoniae bacterium wherein one or more genes encoding a protein according to claim 1 or claim 2 has been rendered inactive. 24. The S.pneumoniae bacterium of claim 23, wherein the gene(s) is/are deleted. 25. The S.pneumoniae bacterium of claim 24, wherein the deletions are isogenic. |
<SOH> BACKGROUND ART <EOH>Streptococcus pneumoniae is a Gram-positive spherical bacterium. It is the most common cause of acute bacterial meningitis in adults and in children over 5 years of age. It is an object of the invention to provide materials for improving the prevention, detection and treatment of Streptococcus pneumoniae infections. More specifically, it is an object of the invention to provide proteins which can be used in the development of vaccines. Further objects are to provide proteins and nucleic acid which can be used in the diagnosis of S.pneumoniae infection, to provide proteins and nucleic acid which can be used for the detection of S.pneumoniae , to provide nucleic acid which is useful for the expression of S.pneumoniae proteins, and to provide proteins which are useful targets for antibiotic research. |
Isolated human transporter proteins, nucleic acid molecules encoding human transporter proteins, and uses thereof |
The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the transporter peptides of the present invention. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the transporter peptides, and methods of identifying modulators of the transporter peptides. |
1. An isolated peptide consisting of an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids. 2. An isolated peptide comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite stand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids. 3. An isolated antibody that selectively binds to a peptide of claim 2. 4. An isolated nucleic acid molecule consisting of a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d). 5. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d). 6. A gene chip comprising a nucleic acid molecule of claim 5. 7. A transgenic non-human animal comprising a nucleic acid molecule of claim 5. 8. A nucleic acid vector comprising a nucleic acid molecule of claim 5. 9. A host cell containing the vector of claim 8. 10. A method for producing any of the peptides of claim 1 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)(d) into a host cell, and culturing the host cell under conditions in which the peptides are expressed from the nucleotide sequence. 11. A method for producing any of the peptides of claim 2 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the peptides are expressed from the nucleotide sequence. 12. A method for detecting the presence of any of the peptides of claim 2 in a sample, said method comprising contacting said sample with a detection agent that specifically allows detection of the presence of the peptide in the sample and then detecting the presence of the peptide. 13. A method for detecting the presence of a nucleic acid molecule of claim 5 in a sample, said method comprising contacting the sample with an oligonucleotide that hybridizes to said nucleic acid molecule under stringent conditions and determining whether the oligonucleotide binds to said nucleic acid molecule in the sample. 14. A method for identifying a modulator of a peptide of claim 2, said method comprising contacting said peptide with an agent and determining if said agent has modulated the function or activity of said peptide. 15. The method of claim 14, wherein said agent is administered to a host cell comprising an expression vector that expresses said peptide. 16. A method for identifying an agent that binds to any of the peptides of claim 2, said method comprising contacting the peptide with an agent and assaying the contacted mixture to determine whether a complex is formed with the agent bound to the peptide. 17. A pharmaceutical composition comprising an agent identified by the method of claim 16 and a pharmaceutically acceptable carrier therefor. 18. A method for treating a disease or condition mediated by a human transorter protein, said method comprising administering to a patient a pharmaceutically effective amount of an agent identified by the method of claim 16. 19. A method for identifying a modulator of the expression of a peptide of claim 2, said method comprising contacting a cell expressing said peptide with an agent, and determining if said agent has modulated the expression of said peptide. 20. An isolated human transporter peptide having an amino acid sequence that shares at least 70% homology with an amino acid sequence shown in SEQ ID NO:2. 21. A peptide according to claim 20 that shares at least 90 percent homology with an amino acid sequence shown in SEQ ID NO:2. 22. An isolated nucleic acid molecule encoding a human transporter peptide, said nucleic acid molecule sharing at least 80 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or 3. 23. A nucleic acid molecule according to claim 22 that shares at least 90 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or 3. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Transporters Transporter proteins regulate many different functions of a cell, including cell proliferation, differentiation, and signaling processes, by regulating the flow of molecules such as ions and macromolecules, into and out, of cells. Transporters are found in the plasma membranes of virtually every cell in eukaryotic organisms. Transporters mediate a variety of cellular functions including regulation of membrane potentials and absorption and secretion of molecules and ion across cell membranes. When present in intracellular membranes of the Golgi apparatus and endocytic vesicles, transporters, such as chloride channels, also regulate organelle pH. For a review, see Greger, R. (1988) Annu. Rev. Physiol. 50:111-122. Transporters are generally classified by structure and the type of mode of action. In addition, transporters are sometimes classified by the molecule type that is transported, for example, sugar transporters, chlorine channels, potassium channels, etc. There may be many classes of channels for transporting a single type of molecule (a detailed review of channel types can be found at Alexander, S. P. H. and J. A. Peters: Receptor and transporter nomenclature supplement. Trends Pharmacol. Sci., Elsevier, pp. 65-68 (1997) and http://www-biology.ucsd.edu/˜msaier/transport/titlepage2.html. The following general classification scheme is known in the art and is followed in the present discoveries. Channel-type transporters. Transmembrane channel proteins of this class are ubiquitously found in the membranes of all types of organisms from bacteria to higher eukaryotes. Transport systems of this type catalyze facilitated diffusion (by an energy-independent process) by passage through a transmembrane aqueous pore or channel without evidence for a carrier-mediated mechanism. These channel proteins usually consist largely of a-helical spanners, although b-strands may also be present and may even comprise the channel. However, outer membrane porin-type channel proteins are excluded from this class and are instead included in class 9. Carrier-type transporters. Transport systems are included in this class if they utilize a carrier-mediated process to catalyze uniport (a single species is transported by facilitated diffusion), antiport (two or more species are transported in opposite directions in a tightly coupled process, not coupled to a direct form of energy other than chemiosmotic energy) and/or symport (two or more species are transported together in the same direction in a tightly coupled process, not coupled to a direct form of energy other than chemiosmotic energy). Pyrophosphate bond hydrolysis-driven active transporters. Transport systems are included in this class if they hydrolyze pyrophosphate or the terminal pyrophosphate bond in ATP or another nucleoside triphosphate to drive the active uptake and/or extrusion of a solute or solutes. The transport protein may or may not be transiently phosphorylated, but the substrate is not phosphorylated. PEP-dependent, phosphoryl transfer-driven group translocators. Transport systems of the bacterial phosphoenolpyruvate:sugar phosphotransferase system are included in this class. The product of the reaction, derived from extracellular sugar, is a cytoplasmic sugar-phosphate. Decarboxylation-driven active transporters. Transport systems that drive solute (e.g., ion) uptake or extrusion by decarboxylation of a cytoplasmic substrate are included in this class. Oxidoreduction-driven active transporters. Transport systems that drive transport of a solute (e.g., an ion>energized by the flow of electrons from a reduced substrate to an oxidized substrate are included in this class. Light-driven active transporters. Transport systems that utilize light energy to drive transport of a solute (e.g., an ion) are included in this class. Mechanically-driven active transporters. Transport systems are included in this class if they drive movement of a cell or organelle by allowing the flow of ions (or other solutes) through the membrane down their electrochemical gradients. Outer-membrane porins (of b-structure). These proteins form transmembrane pores or channels that usually allow the energy independent passage of solutes across a membrane. The transmembrane portions of these proteins consist exclusively of b-strands that form a b-barrel. These porin-type proteins are found in the outer membranes of Gram-negative bacteria, mitochondria and eukaryotic plastids. Methyltransferase-driven active transporters. A single characterized protein currently falls into this category, the Na+-transporting methyltetrahydromethanopterin:coenzyme M methyltransferase. Non-ribosome-synthesized channel-forming peptides or peptide-like molecules. These molecules, usually chains of L- and D-amino acids as well as other small molecular building blocks such as lactate, form oligomeric transmembrane ion channels. Voltage may induce channel formation by promoting assembly of the transmembrane channel. These peptides are often made by bacteria and fungi as agents of biological warfare. Non-Proteinaceous Transport Complexes. Ion conducting substances in biological membranes that do not consist of or are not derived from proteins or peptides fall into this category. Functionally characterized transporters for which sequence data are lacking. Transporters of particular physiological significance will be included in this category even though a family assignment cannot be made. Putative transporters in which no family member is an established transporter. Putative transport protein families are grouped under this number and will either be classified elsewhere when the transport function of a member becomes established, or will be eliminated from the TC classification system if the proposed transport function is disproven. These families include a member or members for which a transport function has been suggested, but evidence for such a function is not yet compelling. Auxiliary transport proteins. Proteins that in some way facilitate transport across one or more biological membranes but do not themselves participate directly in transport are included in this class. These proteins always function in conjunction with one or more transport proteins. They may provide a function connected with energy coupling to transport, play a structural role in complex formation or serve a regulatory function. Transporters of unknown classification. Transport protein families of unknown classification are grouped under this number and will be classified elsewhere when the transport process and energy coupling mechanism are characterized These families include at least one member for which a transport function has been established, but either the mode of transport or the energy coupling mechanism is not known. Ion Channels An important type of transporter is the ion channel. Ion channels regulate many different cell proliferation, differentiation, and signaling processes by regulating the flow of ions into and out of cells. Ion channels are found in the plasma membranes of virtually every cell in eukaryotic organisms. Ion channels mediate a variety of cellular functions including regulation of membrane potentials and absorption and secretion of ion across epithelial membranes. When present in intracellular membranes of the Golgi apparatus and endocytic vesicles, ion channels, such as chloride channels, also regulate organelle pH. For a review, see Greger, R. (1988) Annu. Rev. Physiol. 50:111-122. Ion channels are generally classified by structure and the type of mode of action. For example, extracellular ligand gated channels (ELGs) are comprised of five polypeptide subunits, with each subunit having 4 membrane spanning domains, and are activated by the binding of an extracellular ligand to the channel. In addition, channels are sometimes classified by the ion type that is transported, for example, chlorine channels, potassium channels, etc. There may be many classes of channels for transporting a single type of ion (a detailed review of channel types can be found at Alexander, S.P.H. and J. A. Peters (1997). Receptor and ion channel nomenclature supplement Trends Pharmacol. Sci., Elsevier, pp. 65-68 and http://www-biology.ucsd.edu/˜msaier/transport/toc.html. There are many types of ion channels based on structure. For example, many ion channels fall within one of the following groups: extracellular ligand-gated channels (ELG), intracellular ligand-gated channels (ILG), inward rectifying channels (INR), intercellular (gap junction) channels, and voltage gated channels (VIC). There are additionally recognized other channel families based on ion-type transported, cellular location and drug sensitivity. Detailed information on each of these, their activity, ligand type, ion type, disease association, drugability, and other information pertinent to the present invention, is well known in the art. Extracellular ligand-gated channels, ELGs, are generally comprised of five polypeptide subunits, Unwin, N. (1993), Cell 72: 31-41; Unwin, N. (1995), Nature 373: 37-43; Hucho, F., et al., (1996) J. Neurochem. 66: 1781-1792; Hucho, F., et al., (1996) Eur. J. Biochem. 239: 539-557; Alexander, S.P.H. and J. A. Peters (1997), Trends Pharmacol. Sci., Elsevier, pp.4-6; 36-40; 42-44; and Xue, H. (1998) J. Mol. Evol. 47: 323-333. Each subunit has 4 membrane spanning regions: this serves as a means of identifying other members of the ELG family of proteins. ELG bind a ligand and in response modulate the flow of ions. Examples of ELG include most members of the neurotransmitter-receptor family of proteins, e.g., GABAI receptors. Other members of this family of ion channels include glycine receptors, ryandyne receptors, and ligand gated calcium channels. The Voltage-Gated Ion Channel (VIC) Superfamily Proteins of the VIC family are ion-selective channel proteins found in a wide range of bacteria, archaea and eukaryotes Hille, B. (1992), Chapter 9: Structure of channel proteins; Chapter 20: Evolution and diversity. In: Ionic Channels of Excitable Membranes, 2nd Ed., Sinaur Assoc. Inc., Pubs., Sunderland, Massachusetts; Sigworth, F. J. (1993), Quart. Rev. Biophys. 27: 1-40; Salkoff, L. and T. Jegla (1995), Neuron 15: 489-492; Alexander, S.P.H. et al., (1997), Trends Pharmacol. Sci., Elsevier, pp. 76-84; Jan, L. Y. et al., (1997), Annu. Rev. Neurosci. 20: 91-123; Doyle, D. A, et al., (1998) Science 280: 69-77; Terlau, H. and W. Stühmer (1998), Naturwissenschaften 85: 437-444. They are often homo- or heterooligomeric structures with several dissimilar subunits (e.g., al-a2-d-b Ca 2+ channels, ab 1 b 2 Na + channels or (a) 4 -b K + channels), but the channel and the primary receptor is usually associated with the a (or a1) subunit. Functionally characterized members are specific for K + , Na + or Ca 2+ . The K + channels usually consist of homotetrameric structures with each a-subunit possessing six transmembrane spanners (TMSs). The al and a subunits of the Ca 2+ and Na + channels, respectively, are about four times as large and possess 4 units, each with 6 TMSs separated by a hydrophilic loop, for a total of 24 TMSs. These large channel proteins form heterotetra-unit structures equivalent to the homotetrameric structures of most K + channels. All four units of the Ca 2+ and Na + channels are homologous to the single unit in the homotetrameric K + channels. Ion flux via the eukaryotic channels is generally controlled by the transmembrane electrical potential (hence the designation, voltage-sensitive) although some are controlled by ligand or receptor binding. Several putative K + -selective channel proteins of the VIC family have been identified in prokaryotes. The structure of one of them, the KcsA K + channel of Streptomyces lividans, has been solved to 3.2 Å resolution. The protein possesses four identical subunits, each with two transmembrane helices, arranged in the shape of an inverted teepee or cone. The cone cradles the “selectivity filter” P domain in its outer end. The narrow selectivity filter is only 12 Å long, whereas the remainder of the channel is wider and lined with hydrophobic residues. A large water-filled cavity and helix dipoles stabilize K + in the pore. The selectivity filter has two bound K + ions about 7.5 Å apart from each other. Ion conduction is proposed to result from a balance of electrostatic attractive and repulsive forces. In eukaryotes, each VIC family channel type has several subtypes based on pharmacological and electrophysiological data Thus, there are five types of Ca 2+ channels (L, N, P, Q and T). There are at least ten types of K + channels, each responding in different ways to different stimuli: voltage-sensitive Ka, Kv, Kvr, Kvs and Ksr], Ca 2+ -sensitive [BK Ca , IK Ca and SK Ca ] and receptor-coupled [K M and K ACh ]. There are at least six types of Na + channels (I, II, III, μl, H1 and PN3). Tetrameric channels from both prokaryotic and eukaryotic organisms are known in which each a-subunit possesses 2 TMSs rather than 6, and these two TMSs are homologous to TMSs 5 and 6 of the six TMS unit found in the voltage-sensitive channel proteins. KcsA of S. lividans is an example of such a 2 TMS channel protein. These channels may include the K Na (Na + -activated) and K Vol (cell volume-sensitive) K + channels, as well as distantly related channels such as the Tok1 K + channel of yeast, the TWIK-1 inward rectifier K + channel of the mouse and the TREK-1 K + channel of the mouse. Because of insufficient sequence similarity with proteins of the VIC family, inward rectifier K + IRK channels (ATP-regulated; G-protein-activated) which possess a P domain and two flanking TMSs are placed in a distinct family. However, substantial sequence similarity in the P region suggests that they are homologous. The b, g and d subunits of VIC family members, when present, frequently play regulatory roles in channel activation/deactivation. The Epithelial Na + Channel (ENaC) Family The ENaC family consists of over twenty-four sequenced proteins (Canessa, C. M., et al., (1994), Nature 367: 463-467, Le, T. and M. H. Saier, Jr. (1996), Mol. Membr. Biol. 13: 149-157; Garty, H. and L. G. Palmer (1997), Physiol. Rev. 77: 359-396; Waldmann, R., et al., (1997), Nature 386: 173-177; Darboux, I., et al., (1998), J. Biol. Chem. 273: 9424-9429; Firsov, D., et al., (1998), EMBO J. 17: 344-352; Horisberger, J.-D. (1998). Curr. Opin. Struc. Biol. 10: 443-449). All are from animals with no recognizable homologues in other eukaryotes or bacteria. The vertebrate ENaC proteins from epithelial cells cluster tightly together on the phylogenetic tree: voltage-insensitive ENaC homologues are also found in the brain. Eleven sequenced C. elegans proteins, including the degenerins, are distantly related to the vertebrate proteins as well as to each other. At least some of these proteins form part of a mechano-transducing complex for touch sensitivity. The homologous Helix aspersa (FMRF-amide)-activated Na + channel is the first peptide neurotransmitter-gated ionotropic receptor to be sequenced. Protein members of this family all exhibit the same apparent topology, each with N— and C-termini on the inside of the cell, two amphipathic transmembrane spanning segments, and a large extracellular loop. The extracellular domains contain numerous highly conserved cysteine residues. They are proposed to serve a receptor function. Mammalian ENaC is important for the maintenance of Na + balance and the regulation of blood pressure. Three homologous ENaC subunits, alpha, beta, and gamma, have been shown to assemble to form the highly Na + -selective channel. The stoichiometry of the three subunits is alpha 2 , beta1, gamma1 in a heterotetrameric architecture. The Glutamate-Gated Ion Channel (GIC) Family of Neurotransmitter Receptors Members of the GIC family are heteropentameric complexes in which each of the 5 subunits is of 800-1000 amino acyl residues in length (Nakanishi, N., et al, (1990), Neuron 5: 569-581; Unwin, N. (1993), Cell 72:31-41; Alexander, S.P.H. and J. A. Peters (1997) Trends Pharmacol. Sci., Elsevier, pp.36-40). These subunits may span the membrane three or five times as putative a-helices with the N-termini (the glutamate-binding domains) localized extracellularly and the C-termini localized cytoplasmically. They may be distantly related to the ligand-gated ion channels, and if so, they may possess substantial b-structure in their transmembrane regions. However, homology between these two families cannot be established on the basis of sequence comparisons alone. The subunits fall into six subfamilies: a, b, g, d, e and z. The GIC channels are divided into three types: (1) a-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)-, (2) kainate- and (3) N-methyl-D-aspartate (NMDA)-selective glutamate receptors. Subunits of the AMPA and kainate classes exhibit 35-40% identity with each other while subunits of the NMDA receptors exhibit 22-24% identity with the former subunits. They possess large N-terminal, extracellular glutamate-binding domains that are homologous to the periplasmic glutamine and glutamate receptors of ABC-type uptake permeases of Gram-negative bacteria. All known members of the GIC family are from animals. The different channel (receptor) types exhibit distinct ion selectivities and conductance properties. The NMDA-selective large conductance channels are highly permeable to monovalent cations and Ca 2+ . The AMPA- and kainate-selective ion channels are permeable primarily to monovalent cations with only low permeability to Ca 2+ . The Chloride Channel (ClC) Family The ClC family is a large family consisting of dozens of sequenced proteins derived from Gram-negative and Gram-positive bacteria, cyanobacteria, archaea, yeast, plants and animals (Steinmeyer, K., et al., (1991), Nature 354: 301-304; Uchida, S., et al., (1993), J. Biol. Chem. 268: 3821-3824; Huang, M.-E., et al., (1994), J. Mol. Biol. 242: 595-598; Kawasaki, M., et al, (1994), Neuron 12: 597-604; Fisher, W. E., et al., (1995)i Genomics. 29:598-606; and Foskett, J. K. (1998), Annu. Rev. Physiol. 60: 689-717). These proteins are essentially ubiquitous, although they are not encoded within genomes of Haemophilus influenzae, Mycoplasma genitalium, and Mycoplasma pneumoniae. Sequenced proteins vary in size from 395 amino acyl residues ( M. jannaschii ) to 988 residues (man). Several organisms contain multiple ClC family paralogues. For example, Synechocystis has two paralogues, one of 451 residues in length and the other of 899 residues. Arabidopsis thaliana has at least four sequenced paralogues, (775-792 residues), humans also have at least five paralogues (820-988 residues), and C. elegans also has at least five (810-950 residues). There are nine known members in mammals, and mutations in three of the corresponding genes cause human diseases. E. coli, Methanococcus jannaschii and Saccharomyces cerevisiae only have one ClC family member each. With the exception of the larger Synechocystis paralogue, all bacterial proteins are small (395-492 residues) while all eukaryotic proteins are larger (687-988 residues). These proteins exhibit 10-12 putative transmembrane a-helical spanners (TMSs) and appear to be present in the membrane as homodimers. While one member of the family, Torpedo ClC-O, has been reported to have two channels, one per subunit, others are believed to have just one. All functionally characterized members of the ClC family transport chloride, some in a voltage-regulated process. These channels serve a variety of physiological functions (cell volume regulation; membrane potential stabilization; signal transduction; transepithelial transport, etc.). Different homologues in humans exhibit differing anion selectivities, i.e., ClC4 and ClC5 share a NO 3 − >Cl − >Br − >I − conductance sequence, while ClC3 has an I − >Cl − selectivity. The ClC4 and ClC5 channels and others exhibit outward rectifying currents with currents only at voltages more positive than +20 mV. Animal Inward Rectifier K + Channel (IRK-C) Family IRK channels possess the “minimal channel-forming structure” with only a P domain, characteristic of the channel proteins of the VIC family, and two flanking transmembrane spanners (Shuck, M. E., et al., (1994), J. Biol. Chem. 269: 24261-24270; Ashen, M. D., et al., (1995), Am. J. Physiol. 268: H506-H511; Salkoff, L. and T. Jegla (1995), Neuron 15: 489-492; Aguilar-Bryan, L., et al., (1998), Physiol. Rev. 78: 227-245; Ruknudin, A., et al., (1998), J. Biol. Chem. 273:14165-14171). They may exist in the membrane as homo- or heterooligomers. They have a greater tendency to let K + flow into the cell than out Voltage-dependence may be regulated by external K + , by internal Mg 2+ , by internal AT? and/or by G-proteins. The P domains of IRK channels exhibit limited sequence similarity to those of the VIC family, but this sequence similarity is insufficient to establish homology. Inward rectifiers play a role in setting cellular membrane potentials, and the closing of these channels upon depolarization permits the occurrence of long duration action potentials with a plateau phase. Inward rectifiers lack the intrinsic voltage sensing helices found in VIC family channels. In a few cases, those of Kir1.1a and Kir6.2, for example, direct interaction with a member of the ABC superfamily has been proposed to confer unique functional and regulatory properties to the heteromeric complex, including sensitivity to ATP. The SUR1 sulfonylurea receptor (spQ09428) is the ABC protein that regulates the Kir6.2 channel in response to ATP, and CFTR may regulate kir1.1a. Mutations in SUR1 are the cause of familial persistent hyperinsulinemic hypoglycemia in infancy (PHHI), an autosomal recessive disorder characterized by unregulated insulin secretion in the pancreas. ATP-Gated Cation Channel (ACC) Family Members of the ACC family (also called P2X receptors) respond to ATP, a functional neurotransmitter released by exocytosis from many types of neurons (North, R. A. (1996), Curr. Opin. Cell Biol. 8: 474-483; Soto, F., M. Garcia-Guzman and W. Stühmer (1997), J. Membr. Biol. 160: 91-100). They have been placed into seven groups (P2X 1 -P2X 7 ) based on their pharmacological properties. These channels, which function at neuron-neuron and neuron-smooth muscle junctions, may play roles in the control of blood pressure and pain sensation. They may also function in lymphocyte and platelet physiology. They are found only in animals. The proteins of the ACC family are quite similar in sequence (>35% identity), but they possess 380-1000 amino acyl residues per subunit with variability in length localized primarily to the C-terminal domains. They possess two transmembrane spanners, one about 30-50 residues from their N-termini, the other near residues 320-340. The extracellular receptor domains between these two spanners (of about 270 residues) are well conserved with numerous conserved glycyl and cysteyl residues. The hydrophilic C-termini vary in length from 25 to 240 residues. They resemble the topologically similar epithelial Na + channel (ENaC) proteins in possessing (a) N— and C-termini localized intracellularly, (b) two putative transmembrane spanners, (c) a large extracellular loop domain, and (d) many conserved extracellular cysteyl residues. ACC family members are, however, not demonstrably homologous with them. ACC channels are probably hetero- or homomultimers and transport small monovalent cations (Me + ). Some also transport Ca 2+ ; a few also transport small metabolites. The Ryanodine-Inositol 1,4,5-triphosphate Receptor Ca 2+ Channel (RIR-CaC) Family Ryanodine (Ry)-sensitive and inositol 1,4,5-triphosphate (IP3)-sensitive Ca 2+ -release channels function in the release of Ca 2+ from intracellular storage sites in animal cells and thereby regulate various Ca 2+ -dependent physiological processes (Hasan, G. et al., (1992) Development 116: 967-975; Michikawa, T., et al., (1994), J. Biol. Chem. 269: 9184-9189; Tunwell, R. E. A., (1996), Biochem. J. 318: 477-487; Lee, A. G. (1996) Biomembranes, Vol. 6, Transmembrane Receptors and Channels (A. G. Lee, ed.), JAI Press, Denver, Colo., pp 291-326; Mikoshiba, K., et al., (1996) J. Biochem. Biomem. 6: 273-289). Ry receptors occur primarily in muscle cell sarcoplasmic reticular (SR) membranes, and IP3 receptors occur primarily in brain cell endoplasmic reticular (ER) membranes where they effect release of Ca 2+ into the cytoplasm upon activation (opening) of the channel. The Ry receptors are activated as a result of the activity of dihydropyridine-sensitive Ca 2+ channels. The latter are members of the voltage-sensitive ion channel (VIC) family. Dihydropyridine-sensitive channels are present in the T-tubular systems of muscle tissues. Ry receptors are homotetrameric complexes with each subunit exhibiting a molecular size of over 500,000 daltons (about 5,000 amino acyl residues). They possess C-terminal domains with six putative transmembrane a-helical spanners (TMSs). Putative pore-forming sequences occur between the fifth and sixth TMSs as suggested for members of the VIC family. The large N-terminal hydrophilic domains and the small C-terminal hydrophilic domains are localized to the cytoplasm. Low resolution 3-dimensional structural data are available. Mammals possess at least three isoforms that probably arose by gene duplication and divergence before divergence of the mammalian species. Homologues are present in humans and Caenorabditis elegans. IP 3 receptors resemble Ry receptors in many respects. (1) They are homotetrameric complexes with each subunit exhibiting a molecular size of over 300,000 daltons (about 2,700 amino acyl residues). (2) They possess C-terminal channel domains that are homologous to those of the Ry receptors. (3) The channel domains possess six putative TMSs and a putative channel lining region between TMSs 5 and 6. (4) Both the large N-terminal domains and the smaller C-terminal tails face the cytoplasm. (5) They possess covalently linked carbohydrate on extracytoplasmic loops of the channel domains. (6) They have three currently recognized isoforms (types 1, 2, and 3) in mammals which are subject to differential regulation and have different tissue distributions. IP 3 receptors possess three domains: N-terminal IP 3 -binding domains, central coupling or regulatory domains and C-terminal channel domains. Channels are activated by IP 3 binding, and like the Ry receptors, the activities of the IP 3 receptor channels are regulated by phosphorylation of the regulatory domains, catalyzed by various protein kinases. They predominate in the endoplasmic reticular membranes of various cell types in the brain but have also been found in the plasma membranes of some nerve cells derived from a variety of tissues. The channel domains of the Ry and IP 3 receptors comprise a coherent family that in spite of apparent structural similarities, do not show appreciable sequence similarity of the proteins of the VIC family. The Ry receptors and the IP 3 receptors cluster separately on the RIR-CaC family tree. They both have homologues in Drosophila . Based on the phylogenetic tree for the family, the family probably evolved in the following sequence: (1) A gene duplication event occurred that gave rise to Ry and IP 3 receptors in invertebrates. (2) Vertebrates evolved from invertebrates. (3) The three isoforms of each receptor arose as a result of two distinct gene duplication events. (4) These isoforms were transmitted to mammals before divergence of the mammalian species. The Organellar Chloride Channel (O-ClC) Family Proteins of the O-ClC family are voltage-sensitive chloride channels found in intracellular membranes but not the plasma membranes of animal cells (Landry, D, et al., (1993), J. Biol. Chem. 268: 14948-14955; Valenzuela, Set al., (1997), J. Biol. Chem. 272: 12575-12582; and Duncan, R. R., et al., (1997), J. Biol. Chem. 272: 23880-23886). They are found in human nuclear membranes, and the bovine protein targets to the microsomes, but not the plasma membrane, when expressed in Xenopus laevis oocytes. These proteins are thought to function in the regulation of the membrane potential and in transepithelial ion absorption and secretion in the kidney. They possess two putative transmembrane a-helical spanners (TMSs) with cytoplasmic N— and C-termini and a large luminal loop that may be glycosylated. The bovine protein is 437 amino acyl residues in length and has the two putative TMSs at positions 223-239 and 367-385. The human nuclear protein is much smaller (241 residues). A C. elegans homologue is 260 residues long. Sodium/Glucose Cotransporters The novel human protein, and encoding gene, provided by the present invention is related to the family of sodium/glucose cotransporters, also known as solute carrier family 5. Specifically, the protein/cDNA of the present invention may be an alternative splice form of a protein provided in WO200078953 (see the amino acid sequence alignment in FIG. 2 ). The protein/cDNA of the present invention differs from the art-known protein of WO200078953 in that some of the exons are translated in a different reading frame that generates novel amino acids in the center of the protein of the present invention. The protein of the present invention also shows sequence similarity to rkST1, a sodium/glucose cotransporter isolated from rabbit kidney. rkST1 shares 50-60% amino acid sequence identity with other sodium/glucose cotransporters and was found to be expressed in the brain as well as the kidney (Hitomi et al, Biochim Biophys Acta Mar. 23, 1994;1 190(2):469-72). Sodium/glucose cotransporters play important roles in numerous transport functions including uncoupled passive sodium transport (i.e., sodium uniport), “down-hill” water transport in the absence of substrate, sodium/substrate cotransport; and sodium/substrate/water cotransport (Wright et al., Acta Physiol Scand Suppl August 1998;643:257-64). Due to their importance in human physiology, particularly in regulating numerous transport processes, novel human sodium/glucose transporter proteins/genes, such as provided by the present invention, are valuable as potential targets for the development of therapeutics to treat diseases/disorders caused or influenced by transport defects. Furthermore, SNPs in sodium/glucose transporter genes, such as provided by the present invention, are valuable markers for the diagnosis, prognosis, prevention, and/or treatment of such diseases/disorders. Using the information provided by the present invention, reagents such as probes/primers for detecting the SNPs or the expression of the protein/gene provided herein may be readily developed and, if desired, incorporated into kit formats such as nucleic acid arrays, primer extension reactions coupled with mass spec detection (for SNP detection), or TaqMan PCR assays (Applied Biosystems, Foster City, Calif.). Transporter proteins, particularly members of the sodium/glucose cotransporter subfamily, are a major target for drug action and development Accordingly, it is valuable to the field of pharmaceutical development to identify and characterize previously unknown transport proteins. The present invention advances the state of the art by providing previously unidentified human transport proteins. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is based in part on the identification of amino acid sequences of human transporter peptides and proteins that are related to the sodium/glucose cotransporter subfamily, as well as allelic variants and other mammalian orthologs thereof. These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate transporter activity in cells and tissues that express the transporter. Experimental data as provided in FIG. 1 indicates expression in humans in parathyroid tumors, kidney, and nervous and brain tissue. |
Human secreted proteins |
The present invention relates to human secreted polypeptides, and isolated nucleic acid molecules encoding said polypeptides, useful for diagnosing and treating diabetes mellitus and/or conditions related to diabetes. Antibodies that bind these polypeptides are also encompassed by the present invention. Also encompassed by the invention are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies. The invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The present invention further encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention. |
1. Use of a polypeptide for the preparation of a diagnostic or pharmaceutical composition for diagnosing or treating diabetes or conditions related to diabetes, wherein said polypeptide comprises an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 2. Use of the polypeptide of claim 1, wherein said wherein said polypeptide comprises a heterologous amino acid sequence. 3. Use of a polypeptide for the preparation of a diagnostic or pharmaceutical composition for diagnosing or treating diabetes or conditions related to diabetes, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 4. Use of the polypeptide of claim 3, wherein said polypeptide comprises a heterologous amino acid sequence. 5. Use of an antibody or fragment thereof for the preparation of a diagnostic or pharmaceutical composition for diagnosing or treating diabetes or conditions related to diabetes, wherein said antibody or fragment thereof binds a polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 6. Use of an antibody or fragment thereof for the preparation of a diagnostic or pharmaceutical composition for diagnosing or treating diabetes or conditions related to diabetes, wherein said antibody or fragment thereof binds a polypeptide selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 7. Use of a nucleic acid molecule for the preparation of a diagnostic or pharmaceutical composition for diagnosing or treating diabetes or conditions related to diabetes, wherein said nucleic acid molecule comprises a polynucleotide sequence at least 95% identical to a sequence selected from the group consisting of: (a) a polynucleotide fragment of SEQ ID NO:X as referenced in Table 1A; (b) a polynucleotide encoding a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polynucleotide encoding a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (e) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (f) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (g) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (h) a polynucleotide encoding a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 8. Use of the nucleic acid molecule of claim 7, wherein said nucleic acid molecule comprises a heterologous polynucleotide sequence. 9. Use of a nucleic acid molecule for the preparation of a diagnostic or pharmaceutical composition for diagnosing or treating diabetes or conditions related to diabetes, wherein said nucleic acid molecule comprises a polynucleotide sequence selected from the group consisting of: (a) a polynucleotide fragment of SEQ ID NO:X as referenced in Table 1A; (b) a polynucleotide encoding a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polynucleotide encoding a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (e) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (f) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (g) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (h) a polynucleotide encoding a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 10. Use of the nucleic acid molecule of claim 9, wherein said nucleic acid molecule comprises a heterologous polynucleotide sequence. 11. Use of an agonist or antagonist for the preparation of a pharmaceutical composition for treating diabetes or conditions related to diabetes, wherein said agonist or antagonist binds a polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f). a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 12. Use of an agonist or antagonist for the preparation of a pharmaceutical composition for treating diabetes or conditions related to diabetes, wherein said agonist or antagonist binds a polypeptide selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 13. A polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 14. The polypeptide of claim 13, wherein said polypeptide comprises a heterologous amino acid sequence. 15. Use of the polypeptide of claim 13 for identifying a binding partner comprising: (a) contacting the polypeptide of claim 13 with a binding partner; and (b) determining whether the binding partner increases or decreases activity of the polypeptide. 16. A polypeptide comprising an amino acid sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 17. The polypeptide of claim 16, wherein said polypeptide comprises a heterologous polypeptide sequence. 18. Use of the polypeptide of claim 16 for identifying a binding partner comprising: (a) contacting the polypeptide of claim 16 with a binding partner; and (b) determining whether the binding partner increases or decreases activity of the polypeptide. 19. An antibody or fragment thereof that binds a polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 20. An antibody or fragment thereof that binds a polypeptide selected from the group consisting of: (a) a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (b) a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (e) a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (f) a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (g) a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 21. A nucleic acid molecule comprising a polynucleotide sequence at least 95% identical to a sequence selected from the group consisting of: (a) a polynucleotide fragment of SEQ ID NO:X as referenced in Table 1A; (b) a polynucleotide encoding a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polynucleotide encoding a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (e) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (f) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (g) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (h) a polynucleotide encoding a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 22. The nucleic acid molecule of claim 21, wherein said nucleic acid molecule comprises a heterologous polynucleotide sequence. 23. A recombinant vector comprising the nucleic acid molecule of claim 21. 24. A recombinant vector comprising the nucleic acid molecule of claim 22. 25. A recombinant host cell comprising the recombinant vector of claim 23. 26. A recombinant host cell comprising the recombinant vector of claim 24. 27. A nucleic acid molecule comprising a polynucleotide sequence selected from the group consisting of: (a) a polynucleotide fragment of SEQ ID NO:X as referenced in Table 1A; (b) a polynucleotide encoding a full length polypeptide of SEQ ID NO:Y or a full length polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (c) a polynucleotide encoding a predicted secreted form of SEQ ID NO:Y or a secreted form of the polypeptide encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (d) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A; (e) a polynucleotide encoding a polypeptide fragment of SEQ ID NO:Y or a polypeptide fragment encoded by the cDNA Clone ID in ATCC Deposit No:Z corresponding to SEQ ID NO:Y as referenced in Table 1A, wherein said fragment has biological activity; (f) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 1B; (g) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y as referenced in Table 2; and (h) a polynucleotide encoding a predicted epitope of SEQ ID NO:Y as referenced in Table 1B. 28. The nucleic acid molecule of claim 27, wherein said nucleic acid molecule comprises a heterologous polynucleotide sequence. 29. A recombinant vector comprising the nucleic acid molecule of claim 27. 30. A recombinant vector comprising the nucleic acid molecule of claim 28. 31. A recombinant host cell comprising the recombinant vector of claim 29. 32. A recombinant host cell comprising the recombinant vector of claim 30. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Over the past few decades, an increasing percentage of the population has become diabetic. Diabetes mellitus is categorized into two types: Type I, known as Insulin-Dependent Diabetes Mellitus (IDDM), or Type II, known as Non-Insulin-Dependent Diabetes Mellitus (NIDDM). IDDM is an autoinmmune disorder in which the insulin-secreting pancreatic beta cells of the islets of Langerhans are destroyed. In these individuals, recombinant insulin therapy is employed to maintain glucose homeostasis and normal energy metabolism. NIDDM, on the other hand, is a polygenic disorder with no one gene responsible for the progression of the disease. In NIDDM, insulin resistance eventually leads to the abolishment of insulin secretion resulting in insulin deficiency. Insulin resistance, at least in part, ensues from a block at the level of glucose uptake and phosphorylation in humans. Diabetics demonstrate a decrease in expression in adipose tissue of insulin-receptor substrate 1 (“IRS1”) (Carvalho et al., FASEB J 13(15):2173-8 (1999)), glucose transporter 4 (“GLUT4”) (Garvey et al., Diabetes 41(4):465-75 (1992)), and the novel abundant protein M gene transcript 1 (“apM1”) (Statnick et al., Int J Exp Diabetes 1(2): 81-8 (2000)), as well as other as of yet unidentified factors. Insulin deficiency in NIDDM leads to failure of normal pancreatic beta-cell function and eventually to pancreatic-beta cell death. Insulin affects fat, muscle, and liver. Insulin is the major regulator of energy metabolism. Malfunctioning of any step(s) in insulin secretion and/or action can lead to many disorders, including for example the dysregulation of oxygen utilization, adipogenesis, glycogenesis, lipogenesis, glucose uptake, protein synthesis, thermogenesis, and maintenance of the basal metabolic rate. This malfunctioning results in diseases and/or disorders that include, but are not limited to, hyperinsulinemia, insulin resistance, insulin deficiency, hyperglycemia, hyperlipidemia, hyperketonemia, and diabetes. Numerous debilitating diabetes-related secondary effects include, but are not limited to, obesity, forms of blindness (cataracts and diabetic retinopathy), limb amputations, kidney failure, fatty liver, coronary artery disease, and neuropathy. Some of the current drugs used to treat insulin resistance and/or diabetes (e.g., insulin secratogogues—sulfonylurea, insulin sensitizers—thiazolidenediones and metformin, and alpha-glucosidase and lipase inhibitors) are inadequate due to the dosage amounts and frequency with which they have to be administered as a result of poor pharmacokinetic properties, the lack of effective control over blood sugar levels, and potential side effects, among other reasons. Diabetes Therapeutic proteins in their native state or when recombinantly produced exhibit a rapid in vivo clearance. Typically, significant amounts of therapeutics are required to be effective during therapy. In addition, small molecules smaller than the 20 kDa range can be readily filtered through the renal tubules (glomerulus) leading to dose-dependent nephrotoxicity. Therefore, there is a need for improvement in treatment (e.g., a need for prolonging the effects of therapeutics of diabetes and/or diabetes related conditions). |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention encompasses human secreted proteinsipolypeptides, and isolated nucleic acid molecules encoding said proteinsipolypeptides, useful for detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating diabetes mellitus and conditions related thereto. Antibodies that bind these polypeptides are also encompassed by the present invention; as are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies. The invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The present invention also encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention. detailed-description description="Detailed Description" end="lead"? |
Human secreted proteins |
The present invention relates to human secreted polypeptides, and isolated nucleic acid molecules encoding said polypeptides, useful for diagnosing and treating cancer and other hyperproliferative diseases and disorders. Antibodies that bind these polypeptides are also encompassed by the present invention. Also encompassed by the invention are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies. The invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The present invention further encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention. |
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