text:
A method for obtaining compounds having selected properties for a particular application by forming base modules having at least two structural diversity elements from the reaction of a first compound having at least one structural diversity element and a first reactive group, with a second compound having at least one structural diversity element and a second reactive group, wherein the first and second groups combine by an addition reaction; producing a first array of molecules by varying at least one of the structural diversity elements of the compounds when producing the base modules; and screening the array to determine a first suitable compound for the particular application. The base modules are preferably formed form oxazolone- and aminimide-derived compounds. If desired, the method can be repeated by producing a second array of molecules through the formation of base modules having structural diversity elements that are modified from those of the first suitable compound; and screening the second array of molecules to determine a second suitable compound for the particular application. The second array producing and screening steps can be repeated as often as necessary to achieve an optimum compound for the particular application.
1. A method of identifying a molecule having selected properties for a particular application which comprises the steps of:
a) forming a base module having at least two structural diversity elements by reacting a first compound having at least one structural diversity element and a first reactive group with a second compound having at least one structural diversity element and a second reactive group which is different from said first reactive group, wherein the first and second reactive groups combine by a solution phase addition reaction;
b) producing a first array of at least two different base modules by repeating step a) at least one time while varying at least one of the structural diversity elements of the first or second compounds to produce at least one additional base module having at least two structural diversity elements, at least one of which differs from the structural diversity elements of the base module produced in step a); and
c) simultaneously screening the first array of base modules in accordance with a standard determined by said particular application to identify a first suitable compound for the particular application.
2. The method of
simultaneously screening the second array to determine a second suitable molecule for the particular application.
3. The method of
and a second reactive group, wherein the first and second groups combine by a solution phase addition reaction, and wherein the structural diversity elements are modified from those of the first suitable molecule.
4. The method of
5. The method of
6. The method of
7. The method of
8. The method of
9. The method of
an amino acid derivative;
a nucleotide derivative;
a carbohydrate derivative;
an organic moiety of an alkyl, cycloakyl, aryl, aralkyl or alkaryl group or a substituted or heterocyclic derivative thereof, or of a naturally occurring or synthetic organic structural motif, optionally containing a reporter element, an electrophilic group, a nucleophilic group or a polymerizable group; or
a macromolecular component.
10. The method of
11. The method of
12. The method of
13. The method of
This application is a continuation of U.S. patent application Ser. No. 08/177,497, filed Jan. 5, 1994, now abandoned, the contents of which are hereby incorporated by reference in their entirety.
The present invention relates to a method for the modular development of aminimide- and oxazolone-derived synthetic organic molecules, posessing selected properties for a particular application. This method involves 1.) the synthesis of an array generated from modules of aminimide-forming, oxzolone, oxazolone-forming and/or oxazolone-derived molecules containing a chosen set of substituent groups which confer structural diversity and/or the reaction of these modules with other appropriate reactive groups to produce an array of molecules posessing a chosen set of diverse structural moieties; 2.) the screening of these molecules for the desired set of properties in a target application. The iterative application of this method enables molecules to be produced, having an optimum balance of properties for the particular application.
The discovery of new molecules has traditionally focused in two broad areas, biologically active molecules, which are used as drugs for the treatment of life-threatening diseases, and new materials, which are used in commercial, and especially, in high technological applications. In both areas, the strategy used to discover new molecules has involved two basic operations: (i) a more or less random choice of a molecular candidate, prepared either via chemical synthesis or isolated from natural sources, and (ii) the testing of the molecular candidate for the property or properties of interest. This discovery cycle is repeated indefinitely until a molecule possessing the desirable property "lead molecule", is located. This "lead molecule" discovery process has been inherently ad-hoc in nature and is time-consuming, laborious, unpredictable and costly.
Once a candidate "lead" molecule has been located, the synthetic chemist must subsequently find ways to synthesize structural variants of this lead molecule to optimize its properties in the desired application. In the case where the "lead" molecule is a synthetized organic species or a natural product, the chemist is usually limited to certain structural themes and synthetic reaction schemes. These are dictated largely by the structural composition of the "lead" molecule and by the requirements of the specific application. For example, in cases where the "lead" posesses a functionally important aromatic ring, various electrophillic and nucleophillic substitutions are typically be carried out on the ring to produce variants. Each such case must be approached as a specific independent design and synthesis problem, starting each time from the beginning, because of the lack of availability of an appropriate chemistry to simply alter the structure of the lead compound to produce the variant.
Recently, some attempts have been made to modularize certain synthetic organic reaction schemes to facillitate modification and transformation of a base compound (see, for example, Proc. Natl. Acad. Sci. USA, 90, 6909, 1933). However, the molecules which can be produced by such attempts are extremely limited in their achievable diversity and are still bounded by factors dictated by the choice of specific structural themes. In the case where the "lead molecule" is a naturally occuring biological molecule, such as a peptide, a protein, an oligonucleotide or a carbohydrate, simple synthetic point-modifications to the lead molecule to produce variants are quite difficult to achieve.
A brief account of the strategies and tactics used in the discovery of new molecules is described below. The emphasis is on biologically interesting molecules; however, the technical problems encountered in the discovery of biologically active molecules as outlined here are also illustrative of the problems encountered in the discovery of molecules which can serve as building blocks for the development of new tools and materials for a variety of high technological applications. Furthermore, as discussed below, these problems are also illustrative of the problems encountered in the development of fabricated structures and materials for high technological applications.
Drug Design
Modern theories of biological activity state that biological activities, and therefore physiological states, are the result of molecular recognition events. For example, nucleotides can form complementary base pairs so that complementary single-stranded molecules hybridize resulting in double- or triple-helical structures that appear to be involved in regulation of gene expression. In another example, a biologically active molecule, referred to as a ligand, binds with another molecule, usually a macromolecule referred to as ligand-acceptor (e.g., a receptor or an enzyme), and this binding elicits a chain of molecular events which ultimately gives rise to a physiological state, e.g., normal cell growth and differentiation, abnormal cell growth leading to carcinogenesis, blood-pressure regulation, nerve-impulse-generation and -propagation, etc. The binding between ligand and ligand-acceptor is geometrically characteristic and extraordinarily specific, involving appropriate three-dimensional structural arrangements and chemical interactions.
A currently favored strategy for the development of agents which can be used to treat diseases involves the discovery of forms of ligands of biological receptors, enzymes, or related macromolecules, which mimic such ligands and either boost, i.e., agonize, or suppress, i.e., antagonize, the activity of the ligand. The discovery of such desirable ligand forms has traditionally been carried out either by random screening of molecules (produced through chemical synthesis or isolated from natural sources), or by using a so-called "rational" approach involving identification of a lead-structure, usually the structure of the native ligand, and optimization of its properties through numerous cycles of structural redesign and biological testing. Since most useful drugs have been discovered not through the "rational" approach but through the screening of randomly chosen compounds, a hybrid approach to drug discovery has recently emerged which is based on the use of combinatorial chemistry to construct huge libraries of randomly-built chemical structures which are screened for specific biological activities. (S. Brenner and R. A. Lerner, 1992, Proc. Natl. Acad. Sci. USA 89:53, 81)
Most lead-structures which have been used in the "rational" drug design approach are native polypeptide ligands of receptors or enzymes. The majority of polypeptide ligands, especially the small ones, are relatively unstable in physiological fluids, due to the tendency of the peptide bond to undergo facile hydrolysis in acidic media or in the presence of peptidases. Thus, such ligands are decisively inferior in a pharmacokinetic sense to nonpeptidic compounds, and are not favored as drugs. An additional limitation of small peptides as drugs is their low affinity for ligand acceptors. This phenomenon is in sharp contrast to the affinity demonstrated by large, folded polypeptides, e.g., proteins, for specific acceptors, e.g., receptors or enzymes, which can be in the subnanomolar range. For peptides to become effective drugs, they must be transformed into nonpeptidic organic structures, i.e., peptide mimetics, which bind tightly, preferably in the nanomolar range, and can withstand the chemical and biochemical rigors of coexistence with biological tissues and fluids.
Despite numerous incremental advances in the art of peptidomimetic design, no general solution to the problem of converting a polypeptide-ligand structure to a peptidomimetic has been defined. At present, "rational" peptidomimetic design is done on an ad hoc basis. Using numerous redesign-synthesis-screening cycles, peptidic ligands belonging to a certain biochemical class have been converted by groups of organic chemists and pharmacologists to specific peptidomimetics; however, in the majority of cases results in one biochemical area, e.g., peptidase inhibitor design using the enzyme substrate as a lead, cannot be transferred for use in another area, e.g., tyrosine-kinase inhibitor design using the kinase substrate as a lead.
In many cases, the peptidomimetics that result from a peptide structural lead using the "rational" approach comprise unnatural alpha-amino acids. Many of these mimetics exhibit several of the troublesome features of native peptides (which also comprise alpha-amino acids) and are, thus, not favored for use as drugs. Recently, fundamental research on the use of nonpeptidic scaffolds, such as steroidal or sugar structures, to anchor specific receptor-binding groups in fixed geometric relationships have been described (see for example Hirschmann, R. et al., 1992 J. Am. Chem. Soc., 114:9699–9701; Hirschmann, R. et al., 1992 J. Am. Chem. Soc., 114:9217–9218); however, the success of this approach remains to be seen.
In an attempt to accelerate the identification of lead-structures, and also the identification of useful drug candidates through screening of randomly chosen compounds, researchers have developed automated methods for the generation of large combinatorial libraries of peptides and certain types of peptide mimetics, called "peptoids", which are screened for a desirable biological activity. For example, the method of H. M. Geysen, (1984 Proc. Natl. Acad. Sci. USA 81:3998) employs a modification of Merrifield peptide synthesis, wherein the C-terminal amino acid residues of the peptides to be synthesized are linked to solid-support particles shaped as polyethylene pins; these pins are treated individually or collectively in sequence to introduce additional amino-acid residues forming the desired peptides. The peptides are then screened for activity without removing them from the pins. Houghton, (1985, Proc. Natl. Acad. Sci. USA 82:5131; and U.S. Pat. No. 4,631,211) utilizes individual polyethylene bags ("tea bags") containing C-terminal amino acids bound to a solid support. These are mixed and coupled with the requisite amino acids using solid phase synthesis techniques. The peptides produced are then recovered and tested individually. Fodor et al., (1991, Science 251:767) described light-directed, spatially addressable parallel-peptide synthesis on a silicon wafer to generate large arrays of addressable peptides that can be directly tested for binding to biological targets. These workers have also developed recombinant DNA/genetic engineering methods for expressing huge peptide libraries on the surface of phages (Cwirla et al., 1990, Proc. Natl. Acad. Sci. USA 87:6378).
In another combinatorial approach, V. D. Huebner and D. V. Santi (U.S. Pat. No. 5,182,366) utilized functionalized polystyrene beads divided into portions each of which was acylated with a desired amino acid; the bead portions were mixed together, then divided into portions each of which was re-subjected to acylation with a second desirable amino acid producing dipeptides, using the techniques of solid phase peptide synthesis. By using this synthetic scheme, exponentially increasing numbers of peptides were produced in uniform which were then separately screened for a biological activity of interest.
Zuckerman et al., (1992, Int. J. Peptide Protein Res. 91:1) also have developed similar methods for the synthesis of peptide libraries and applied these methods to the automation of a modular synthetic chemistry for the production of libraries of N-alkyl glycine peptide derivatives, called "peptoids", which are screened for activity against a variety of biochemical targets. (See also, Symon et al., 1992, Proc. Natl. Acad. Sci. USA 89:9367). Encoded combinatorial chemical syntheses have been described recently (S. Brenner and R. A. Lerner, 1992, Proc. Natl. Acad. Sci. USA 89:5381).
The focus of these structural diversity activities on peptide synthesis chemistry is a direct result of the fact that the ability to generate structural diversity requires, as its starting point, the access to practical stepwise sequential synthesis chemistries which allow the incorporation of varied structural elements with orthagonal reactivities. To-date, these have only been worked out for the Merrifield synthesis of peptides and the Carruthers synthesis of oligonucleotides. Thus, there remains a need for an improved method for the structure-directed generation and screening of organic compounds to determine which may be suitable in a particular application.
The invention relates to a method for obtaining compounds having selected properties for a particular application by forming base modules having at least two structural diversity elements from the reaction of a first compound having at least one structural diversity element and a first reactive group, with a second compound having at least one structural diversity element and a second reactive group, wherein the first and second groups combine by an addition reaction; producing a first array of molecules by varying at least one of the structural diversity elements of the compounds when producing the base modules; and screening the array to determine a first suitable compound for the particular application.
If desired, the method can be repeated by producing a second array of molecules through the formation of base modules having structural diversity elements that are modified from those of the first suitable compound; and screening the second array of molecules to determine a second suitable compound for the particular application. The second array can be produced by forming base modules having at least two structural diversity elements in the same manner as the first array, except that the structural diversity elements are modified from those of the first suitable compound. The second array producing and screening steps can be repeated as often as necessary to achieve an optimum compound for the particular application.
Preferably, the first compound is produced by forming an oxazolone compound having at least one structural diversity element attached thereto and reacting it with a nucleophile or carbonyl compound which contains at least one structural diversity element to form a base module having one of the following structures:
Alternatively, it is also preferred to provide the first compound as an aminimide-forming compound having at least one structural diversity element attached thereto and to react it with an oxazolone or ether compound which contains at least one structural diversity element to form a base module having one of the following structures:
Advantageously, the first and second structural diversity elements can be one of the following:
an amino acid derivative of the form (AA)n;
a nucleotide derivative of the form (NUCL)n;
a carbohydrate derivative of the form (CH)n;
an organic moiety of an alkyl, cycloalkyl, aryl, aralkyl or alkaryl group or a substituted or heterocyclic derivative thereof, or of a naturally occurring or synthetic organic structural motif, optionally containing a reporter element, an electrophilic group, a nucleophilic group or a polymerizable group; or
a macromolecular component.
If desired, at least one of the first and second compounds can be provided with two or more structural diversity elements, two of which can form a ring structure. The invention relates to a method for obtaining compounds having selected properties for a particular application by producing a first structurally diverse array of molecules having at least two orthogonal reactivity elements wherein a first orthogonal reactivity element is held constant for each molecule and a second orthogonal reactivity element is varied; screening the array to determine a first suitable compound for the intended application; and modifying the first suitable compound to form a second structurally diverse array of molecules. Preferably, the first suitable compound has at least two orthogonal reactivity elements, so that the method further includes modifying the first suitable compound by holding a first orthogonal reactivity element constant while varying the second orthogonal reactivity element to produce the second structurally diverse array; and screening the second structurally diverse array of molecules to determine a second suitable compound for the intended application. The method further comprises repeating the modifying and screening steps as often as necessary to achieve the optimum compound for the intended application.
The first structurally diverse array of molecules is advantageously produced by reacting either an oxazolone or aminimide compound, or combinations thereof, with first and second components which provide the orthogonal reactivity elements. It is useful for the first structurally diverse array of molecules to have one of the specific structures disclosed herein. These structures may include components such as an amino acid derivative, a nucleotide derivative, a carbohydrate derivative, an organic structural motif, a reporter element, a polymerizable organic moeity, or a macromolecular component.
This method is useful for a wide variety of applications, including the development of new biopharmaceutical agents, new monomeric species for the modular construction of separations tools, including chiral selectors, industrial detergents and additives and for the development of modular chemical intermediates for the production new materials and polymers. Specifically, the method relates to the selection of molecular modules containing appropriate structural diversity elements, the connecting of these modules together via facile high-yield addition reactions which produce discrete highly pure molecules in less than milligram quantities, in a manner such that the properties of these molecules are determined by the contributions of the individual building modules. The molecular modules of the invention may be chiral, and can be used to synthesize new compounds, structures and materials which are able to recognize biological receptors, enzymes, genetic materials, and other chiral molecules, and are thus of great interest in the fields of biopharmaceuticals, separation industrial and materials science.
The present invention is able to generate a number of different molecules for screening purposes by first forming a base module that contains at least two structural diversity elements attached thereto. These modules are formed by reacting first and second compounds, each of which has at least one structural diversity element and a reactive group. The reactive groups of the first and second compounds are such that they react with each other to form the base module by an additional reaction. By fixing one of the positions and structures of the structural diversity elements and by varying at least one of the others, an array of different molecules is easily generated. These molecules can then be screened to determine which are suitable for a particular application or target use. Once a suitable compound is identified, it can be selected for generating a further array of molecules. This is done by modifying the particular structural diversity elements that are found to be suitable, or by combining the chosen structural diversity element with an expanded or different set of second compounds or elements. This process can be repeated as often as necessary to develop the optimum compound for the particular use.
The particular base module chosen for use in accordance with the present invention is not critical and can be any one of a wide variety of structures. It has been found, however, that two particular structures which are known in the art are highly useful as such base modules, these known compounds being the oxazolones and aminamides. Thus, it is preferred to utilize compounds which are aminamide forming, oxazolone forming, oxazolone or oxazolone-derived molecules for use as the base module. Depending upon the specific structure selected, these base modules can have between two and six structural diversity elements. The specific chemistry of these molecules, as well as an identification of the structural diversity elements and reactivity groups, follows.
Oxazolones, or azlactones, are structures of the general formula:
Oxazolones may posess two substituents at the 4-position. When these substituents are not equivalent, the carbon atom at the 4-position is asymmetric and two non-superimposable oxazolone structures (azlactones) result:
Chiral oxazolones possessing a single 4-position non hydrogen substituent (also known as 5(4H)-oxazolones), derived from (chiral) natural amino acid derivatives, including activated acylamino acyl structures, have been prepared and isolated in the pure, crystalline state (Bodansky, M.; Klausner, Y. S.; Ondetti, M. A. in "Peptide Synthesis", Second Edition, John Wiley & Sons, New York, 1976, p. 14 and references cited therein). The facile, base-catalyzed racemization of several of these oxazolones has been studied in connection with investigations of the serious racemization problem confronting peptide synthesis (see Kemp, D. S. in "The Peptides, Analysis, Synthesis, and Biology", Vol. 1, Gross, E. & Meienhofer, J. editors, 1979, p. 315).
Racemization during peptide synthesis becomes very extensive when the desired peptide is produced by aminolysis of activated peptidyl carboxyl, as in the case of peptide chain extension from the amino terminus, e.g. I–VI shown below (see Atherton, E.; Sheppard, R. C. "Solid Phase Peptide Synthesis, A Practical Approach", IRL Press at Oxford University Press, 1989, pages 11 and 12). An extensively studied mechanism describing this racemization involves conversion of the activated acyl derivative (II) to an oxazolone (III) followed by facile base-catalyzed racemization of the oxazolone via a resonance-stabilized intermediate (IV) and aminolysis of the racemic oxazolone (V) producing racemic peptide products (VI).
Extensive research on the trapping of oxazolones III (or of their activated acyl precursors II) to give acylating agents which undergo little or no racemization upon aminolysis has been carried out, and successes in this area (such as the use of N-hydroxybenzotriazole) have greatly advanced the art of peptide synthesis (Kemp, D. S. in "The Peptides, Analysis, Synthesis, and Biology", Vol. 1, Gross, E. & Meienhofer, J. editors, 1979, p. 315).
Thus, attempts to deal with the racemization problem in peptide synthesis have involved suppressing or avoiding the formation of oxazolone intermediates altogether.
Oxazolones having at least one hydrogen substituent at the 4-position can also undergo a variety of rearrangements and side-reactions (cf., 23 Tetrahedron 3363 (1967)), which may interfere with other desired transformations. This is illustrated for the case of the oxazolone formed from the cyclization of N-acryloyl glycine:
Oxazolones containing two non-hydrogen substituents at the four position are structurally precluded from undergoing these racemizations and side-reactions. These disubstituted oxazolones may be obtained chirally pure and may be subjected to the transformations which are the subject of this invention with retention of the chirality at this position.
When the substituent at the 2-position is capable of undergoing addition reactions, these may be carried out with retention of the chirality at the 4-position to produce new oxazolones. This is shown for the Michael-type addition to an alkenyl oxazolone as follows:
Oxazolones may be prepared from the appropriate amino acid using any of a number of standard acylation and cyclization techniques well-known to those skilled in the art, e.g.:
These oxazolones may be isolated in the pure state or may be generated in-situ from the acyl amino acid by treatment, for example, with equivalent amounts of triethyl amine and ethyl chloroformate in benzene. Following the evolution of carbon monoxide and the removal of the triethyl ammonium chloride formed by filtration. the solution of the oxazolone may be utilized directly for subsequent transformations.
Reactions of Oxazolones
Ring-opening Addition
Oxazolones may be subjected to ring opening reactions with a variety of nucleophiles, as shown below:
In the structure above, Y represents an oxygen, sulfur, or nitrogen atom. R1 and R2 differ from one another and taken alone each signifies one of the following: alkyl including carbocyclic and substituted forms thereof; aryl, aralkyl, alkaryl, and substituted or heterocyclic versions thereof.
The above ring-opening reaction can be carried out either in an organic solvent such as methylene chloride, ethyl acetate, dimethyl formamide (DMF) or in water at room or higher temperatures, in the presence or absence of acids, such as carboxylic, other proton or Lewis-acids, or bases, such as tertiary amines or hydroxides, serving as catalysts.
This reaction may be used to generate an array of adducts, posessing combinations of the structural diversity elements A and C, as shown:
In addition, by appropriate selection of the R and R′ groups, two additional diversity elements can be provided in those positions. Thus, the compound shown can have from two to four structural diversity elements attached to the base module as desired.
Carbonyl Addition
When both substituents in the 4-position are hydrogen, i.e., the oxazolone is formed from cyclization of an acyl glycine, the ring may undergo a high yield condensation addition reaction with aldehyde or ketone-containing structural groups at the 4-position. This reaction may be used to generate an array of adducts, posessing combinations of the structural diversity elements A and B, as shown:
Again, as noted above, the R group can be selected to be a diversity element to provide an additional structural diversity group on the oxazolone molecule.
Combination of the Two Reactions
The resulting adduct may subsequently undergo a high yield ring-opening addition reactrion with a wide variety of SH, NH and OH containing nucleophiles. This reaction sequence may, thus, be used to generate an array of adducts, posessing combinations of the structural diversity elements A, B and C, as shown
Again, as noted above, the R group can be selected to be a diversity element to provide an additional structural diversity group on the oxazolone molecule.
This is illustrated for the case of the in-situ generation of the oxazolone from hippuric acid, followed by removal of the triethylammonium chloride by filtration, the addition of benzaldehyde to form the unsaturated adduct and the ring opening addition of benzylamine to give the tris-phenyl substituted adduct shown:
The ability of these various reactions to be carried out in a stepwise sequential manner using modules chosen in a structure-directed manner allows the production of structurally directed thematic diversity libraries, having structural elements systematically varied around a basic motif.
Aminimides are zwitterionic structures described by the resonance hybrid of the two energetically comparable Lewis structures shown below:
The tetrasubstituted nitrogen of the aminimide group can be asymetric rendering aminimides chiral as shown by the two enantiomers below:
As a result of the polarity of their structures, but lack of net charge, simple aminimides are freely soluble in both water and (especially) organic solvents.
Dilute aqueous solutions of aminimides are neutral and of very low conductivity; the conjugate acids of simple aminimides are weakly acidic, pKa of ca. 4.5. A striking property of aminimides is their hydrolytic stability, under acidic, basic, or enzymatic conditions. For example, boiling trimethyl amine benzamide in 6 N NaOH for 24 hrs leaves the aminimide unchanged. Upon thermolytic treatment, at temperatures exceeding 180� C., aminimides decompose to give isocyanates as follows.
Aminimides can be synthesized in a variety of different ways. The compounds of the present invention can be synthesized by many routes. It is well known in the art of organic synthesis that many different synthetic protocols can be used to prepare a given compound. Different routes can involve more or less expensive reagents, easier or more difficult separation or purification procedures, straightforward or cumbersome scale-up, and higher or lower yield. The skilled synthetic organic chemist knows well how to balance the competing characteristics of competing strategies. Thus, the compounds of the present invention are not limited by the choice of synthetic strategy and any synthetic strategy that yields the compounds described above can be used.
Aminimides via Alkylation of N,N-Disubstituted Hydrazides
Alkylation of a hydrazide followed by neutralization with a base produces an aminimide
Acylation of a suitable trialkyl hydrazinium salt by an acyl derivative or isocyanate in the presence of a strong base in a suitable organic solvent, e.g. dioxane, ether, acetonitrile, etc. produces good yields of aminimides.
A very useful and versatile synthesis of aminimides involves the one-pot reaction of an epoxide, an asymetrically disubstituted hydrazine, and an ester in a hydroxylic solvent, usually water or an alcohol, which is allowed to proceed usually at room temperature over several hours to several days.
The rates for the above reaction increase with increasing electrophilicity of the ester component. Generally, a mixture of 0.1 mol of each of the reactants in 50–100 ml of an appropriate solvent is stirred for the required period at room temperature (the reaction may be monitored by thin layer chromatography). At the end of this period, the solvent is removed in vacuo to give the crude product.
Any of the various R groups illustrated in all of these aminimide and aminimide-forming structures may be selected to be structural diversity elements.
The ability of these various reactions to be carried out using modules chosen in a structure-directed manner allows the production of structurally directed thematic diversity libraries, having structural elements systematically varied around a basic motif.
Further details on the reaction possibilities for the oxazolone and aminimide compounds can be found in two PCT applications PCT/US93/12591 and PCT/US93/12612, each filed on Dec. 28, 1993, and entitled Modular Design And Synthesis of Oxazolone-Derived Molecules and Modular Design And Synthesis Of Aminimide-Derived Molecules, respectively. The content of each of those applications is expressly incorporated herein by reference thereto to the extent necessary to understand the metes and bounds of this invention.
Mixed Aminimide-Oxazolones
A particularly useful embodiment of the invention is the synthesis of mixed aminimide-oxazolone molecules, as shown below. This scenario allows the incorporation of multiple structural diversity elements as shown:
Again, as noted above, the R and methyl groups can be replaced with additional structural diversity elements so that a total of six can be provided on the mixed aminamide-oxazolone base module.
Structural Diversity Elements
Any of a wide variety of structural diversity elements can be used. These elements would include:
1) Amino acid derivatives of the form (AA)N, which would include, for example, natural and synthetic amino acid residues (N=1) including all of the naturally occuring alpha amino acids, especially alanine, arginine, asparagnine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine; the naturally occuring disubstituted amino acids, such as amino isobutyric acid, and isovaline, etc.; a variety of synthetic amino acid residues, including alpha-disubstituted variants, species with olefinic substitution at the alpha position, species having derivatives, variants or mimetics of the naturally occuring side chains; N-Substituted glycine residues; natural and synthetic species known to functionally mimic amino acid residues, such as statine, bestatin, etc. Peptides (N=2–30) constructed from the amino acids listed above, such as angiotensinogen and its family of physiologically important angiotensin hydrolysis products, as well as derivatives, variants and mimetics made from various combinations and permutations of all the natural and synthetic residues listed above. Polypeptides (N=31–70), such as big endothelin, pancreastatin, human growth hormone releasing factor and human pancreatic polypeptide. Proteins (N>70) including structural proteins such as collagen, functional proteins such as hemoglobin, regulatory proteins such as the dopamine and thiombin receptors.
2) Nucleotide derivatives of the form (NUCL)N, which includes natural and synthetic nucleotides (N=1) such as adenosine, thymine, guanidine, uridine, cystosine, derivatives of these and a variety of variants and mimetics of the purine ring, the sugar ring, the phosphate linkage and combinations of some or all of these. Nucleotide probes (N=2–25) and oligonucleotides (N>25) including all of the various possible homo and heterosynthetic combinations and permutations of the naturally occuring nucleotides, derivatives and variants containing synthetic purine or pyrimidine species or mimics of these, various sugar ring mimetics, and a wide variety of alternate backbone analogues including but not limited to phosphodiester, phosphorothionate, phosphorodithionate, phosphoramidate, alkyl phosphotriester, sulfamate, 3′-thioformacetal, methylene(methylimino), 3-N-carbamate, morpholino carbamate and peptide nucleic acid analogues.
3) Carbohydrate derivatives of the form (CH)n. This would include natural physiologically active carbohydrates such as including related compounds such as glucose, galactose, sialic acids, beta-D-glucosylamine and nojorimycin which are both inhibitors of glucosidase, pseudo sugars, such as 5a-carba-2-D-galactopyranose, which is known to inhibit the growth of Klebsiella pneumonia (n=1), synthetic carbohydrate residues and derivatives of these (n=1) and all of the complex oligomeric permutations of these as found in nature, including high mannose oligosaccharides, the known antibiotic streptomycin (n>1).
4) A naturally occurring or synthetic organic structural motif. This term is defined as meaning an organic molecule having a specific structure that has biological activity, such as having a complementary structure to an enzyme, for instance. This term includes any of the well known base structures of pharmaceutical compounds including pharmacophores or metabolites thereof. These include beta-lactams, such as pennicillin, known to inhibit bacterial cell wall biosynthesis; dibenzazepines, known to bind to CNS receptors, used as antidepressants; polyketide macrolides, known to bind to bacterial ribosymes, etc. These structural motifs are generally known to have specific desirable binding properties to ligand acceptors.
5) A reporter element such as a natural or synthetic dye or a residue capable of photographic amplification which possesses reactive groups which may be synthetically incorporated into the oxazolone structure or reaction scheme and may be attached through the groups without adversely interfering with the reporting functionality of the group. Preferred reactive groups are amino, thio, hydroxy, carboxylic acid, carboxylic acid ester, particularly methyl ester, acid chloride, isocyanate alkyl halides, aryl halides and oxirane groups.
6) An organic moiety containing a polymerizable group such as a double bond or other functionalities capable of undergoing condensation polymerization or copolymerization. Suitable groups include vinyl groups, oxirane groups, carboxylic acids, acid chlorides, esters, amides, lactones and lactams. Other organic moiety such as those defined for R and R′ may also be used.
7) A macromolecular component, such as a macromolecular surface or structures which may be attached to the oxazolone modules via the various reactive groups outlined above in a manner where the binding of the attached species to a ligand-receptor molecule is not adversely affected and the interactive activity of the attached functionality is determined or limited by the macromolecule. This includes porous and non-porous inorganic macromolecular components, such as, for example, silica, alumina, zirconia, titania and the like, as commonly used for various applications, such as normal and reverse phase chromatographic separations, water purification, pigments for paints, etc.; porous and non-porous organic macromolecular components, including synthetic components such as styrene-divinyl benzene beads, various methacrylate beads, PVA beads, and the like, commonly used for protein purification, water softening and a variety of other applications, natural components such as native and functionalized celluloses, such as, for example, agarose and chitin, sheet and hollow fiber membranes made from nylon, polyether sulfone or any of the materials mentioned above. The molecular weight of these macromolecules may range from about 1000 Daltons to as high as possible. They may take the form of nanoparticles (dp=100–1000 Angstroms), latex particles (dp=1000–5000 Angstroms), porous or non-porous beads (dp=0.5–1000 microns), membranes, gels, macroscopic surfaces or functionalized or coated versions or composites of these.
8) A structural moiety selected from the group including cyano, nitro, halogen, oxygen, hydroxy, alkoxy, thio, straight or branched chain alkyl, carbocyclic aryl and substituted or heterocyclic derivatives thereof, wherein R and R′ may be different in adjacent n units and have a selected stereochemical arrangement about the carbon atom to which they are attached;
As used herein, the phrase linear chain or branched chained alkyl groups means any substituted or unsubstituted acyclic carbon-containing compounds, including alkanes, alkenes and alkynes. Alkyl groups having up to 30 carbon atoms are preferred. Examples of alkyl groups include lower alkyl, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl or tert-butyl; upper alkyl, for example, cotyl, nonyl, decyl, and the like; lower alkylene, for example, ethylene, propylene, propyldiene, butylene, butyldiene; upper alkenyl such as 1-decene, 1-nonene, 2,6-dimethyl-5-octenyl, 6-ethyl-5-octenyl or heptenyl, and the like; alkynyl such as 1-ethynyl, 2-butynyl, 1-pentynyl and the like. The ordinary skilled artisan is familiar with numerous linear and branched alkyl groups, which are within the scope of the present invention.
In addition, such alkyl group may also contain various substituents in which one or more hydrogen atoms has been replaced by a functional group. Functional groups include but are not limited to hydroxyl, amino, carboxyl, amide, ester, ether, and halogen (fluorine, chlorine, bromine and iodine), to mention but a few. Specific substituted alkyl groups can be, for example, alkoxy such as methoxy, ethoxy, butoxy, pentoxy and the like, polyhydroxy such as 1,2-dihydroxypropyl, 1,4-dihydroxy-1-butyl, and the like; methylamino, ethylamino, dimethylamino, diethylamino, triethylamino, cyclopentylamino, benzylamino, dibenzylamino, and the like; propanoic, butanoic or pentanoic acid groups, and the like; formamido, acetamido, butanamido, and the like, methoxycarbonyl, ethoxycarbonyl or the like, chloroformyl, bromoformyl, 1,1-chloroethyl, bromo ethyl, and the like, or dimethyl or diethyl ether groups or the like.
As used herein, substituted and unsubstituted carbocyclic groups of up to about 20 carbon atoms means cyclic carbon-containing compounds, including but not limited to cyclopentyl, cyclohexyl, cycloheptyl, admantyl, and the like. such cyclic groups may also contain various substituents in which one or more hydrogen atoms has been replaced by a functional group. Such functional groups include those described above, and lower alkyl groups as described above. The cyclic groups of the invention may further comprise a heteroatom. For example, in a specific embodiment, R2 is cycohexanol.
As used herein, substituted and unsubstituted aryl groups means a hydrocarbon ring bearing a system of conjugated double bonds, usually comprising an even number of 6 or more (pi) electrons. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anisyl, toluyl, xylenyl and the like. According to the present invention, aryl also includes aryloxy, aralkyl, aralkyloxy and heteroaryl groups, e.g., pyrimidine, morpholine, piperazine, piperidine, benzoic acid, toluene or thiophene and the like. These aryl groups may also be substituted with any number of a variety of functional groups. In addition to the functional groups described above in connection with substituted alkyl groups and carbocylic groups, functional groups on the aryl groups can be nitro groups.
As mentioned above, these structural moieties can also be any combination of alkyl, carbocyclic or aryl groups, for example, 1-cyclohexylpropyl, benzylcyclohexylmethyl, 2-cyclohexyl-propyl, 2,2-methylcyclohexylpropyl, 2,2methylphenylpropyl, 2,2-methylphenylbutyl, and the like.
Orthogonal Reactivities
A key element of the present method is the presence of at least two compounds, each having a reactive group capable of forming an addition compound with the other and carrying at least one of the structural diversity groups. These compounds are used to form the aminimide and the oxazolone base modules. These compounds may take the form of either A.) multiple reactive groups which are capable of being "turned on" independently of each other or B.) groups with multiple states with differing reactivities which may be addressed or brought into being at different times or under different conditions in a reaction sequence. It is highly desirable, although not absolutely necessary, that each individual reaction be a high-yielding addition reaction without possible interfering side-reactions so that isolation and purification steps are not necesary, or, at least, are held to a minimum.
Specifically preferred reactive groups to generate the aminimide and oxazolone structures and the resulting base modules are listed below in tables 1, 2 and 3. The bonds in the structures in these figures represent potential points of attachment for the attachment of the structural diversity elements to the first and second compounds and to the base modules.
This example describes the generation of a matrix of 16 molecules around the following aryl-heterocycle-alicyclic amine structural theme.
The 2-phenyl and 2-(2-naphthyl)-5-oxazolones (produced by reacting the lithium salt of glucine with the aryl acid chlorides, followed by cyclization with ethyl chloroformate at 0 C) were reacted with 2-furfural, 3-fufural, 2-thiophenal and 3-thiophenyl to produce the oxazolones functionalized at the 4-position. This was followed by subsequent ring-opening addition of 4-(3-aminopropylmorpholine and 1-(3-aminopropyl)-2-pipicoline to form the adducts shown. The reactions were carried out in individual vials such that each vial contained one pure final compound as follows:
1.) equimolar quantities of the oxazolone and the aldehyde dissolved in dry benzene (25 ml/gm reactants) were heated to 75 C for 15 minutes; 2.) the reaction mixture was cooled to 10 C and the amine was added dropwise with stirring; 3.) the mixture was re-heated to 75 C for 20 minutes and 4.) the solvent was removed in vacuo to give the crude solid product.
The following example outlines the generation of a matrix of 16 molecules around the basic structural theme of a hydroxy-proline transition state mimetic inhibitor for proteases:
Structural Theme:
These 16 materials were isolated in essentially quantitative yield on removal of the reaction solvent by evaporation and purified samples were obtained as crystalline solids after recrystallization from ethyl acetate and characterized by 1H-NMR, FTIR and other analytical techniques. The set of molecules where X=CH2 was tested as competitive inhibitiors of the enzyme chymotrypsin in a standard assay using a BTEE substrate. The results found for Ki were 200 uM for R1=Ph, R2=Me; 130 uM for R1=Me, R2=Ph; 500 uM for R1=Ph, R2=Ph; and R1=Me, R2=Me was found to not be an inhibitor. These results indicate a preference of the enzyme in this assay for one phenyl and one methyl, with the phenyl being preferred in the R1 position. Based on these results, a second array was synthesized using phenyl groups in this position having a variety of different substituent groups for further testing against the enzyme.
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text:
Popular among both children and adults, the line started with basic frogs, dolphins and bears. Over time, it grew to include seasonal animals and celebrity lookalikes, like the Princess Diana bear introduced in 1997 after the royal's death.
Bamsey said the craze-inducing creatures fit the mold of a fad but said the shorter life cycle is part of its offering.
"Creating and selling 'fads' can be a wise business choice if the category/product lends itself to that," he said. "It's all about generating interest and intent to buy. And some products are unlikely to be sustainable over time."
Since Ty Inc. is a low-profile, private company, sales figures are difficult to come by. But while the company still sells the plush toys, it no longer grabs headlines and inspires buying frenzies the way they used to. However, now that a Beanie Baby fashioned after the President Obama's dog Bo is in the works, maybe they're poised for a comeback.
The Macarena
A favorite at weddings and bar mitzvahs once upon a time, Los del Rio's Macarena is No. 5 on the Billboard Hot 100 All-Time Top Songs list
The popular song -- and unforgettable accompanying line dance -- peaked during the summer of 1996. But after the song's success, the band fell out of the mainstream, winning the top spot on VH1's top one-hit wonders list in 2002.
In 2004, the social networking Web site was a media industry darling, making it on to Time magazine's list of the 50 Coolest Web sites that year. But those days are long gone.
It's still popular in some parts of the world, analysts said, but its popularity in the United States has plummeted as other sites like Facebook and Twitter have taken its place.
"Friendster, the original social network, exploded then imploded. Facebook worked out the errors made during that wave and took it all to another level altogether," said PSFK's Fawkes.
The site was launched in 2003, giving users a place to connect with existing friends, friends of friends, their friends and more. By June 2005, the site claimed more than 16 million worldwide subscribers.
According to its Web site, Friendster now has about 100 million members worldwide, but it isn't even listed among the top 10 social networking sites. According to Compete, Friendster was the 18th most-visited social networking site in January 2009, with about 7 million visits. Facebook, ranked No. 1, had about 1.2 billion visits in the same month.
Ring Pops
"The quintessential fad," said Ira Matathia of Faith Popcorn's BrainReserve, about the pacifier-like ring pops.
Manufactured by the Topps Company, the wearable sweets were introduced in 1977, but they really became popularized in the '80s. The edible jewelry is tough to find these days, though it still makes appearances on the fingers of stars like Lindsay Lohan, Fergie and others.
Garbage Pail Kids
Another Topps creation, released in 1985, Garbage Pail Kids were once the trading card of choice for kids and adolescents.
Meant to parody the Cabbage Patch Kids dolls, each card featured a character that had suffered some kind of abnormality. The cards' success led to the production of a live-action movie and animated television series that briefly aired in Europe.
The fervor has died down, but the cards are still around. Its Web site gives visitors the option to build and raiser their own kids.

subset: commoncrawl


text:
It looks like you're new here. If you want to get involved, click one of these buttons!
We make various files available for public download from the GSA FTP server, such as the GATK resource bundle and presentation slides. We also maintain a public upload feature for processing bug reports from users.
There are two logins to choose from depending on whether you want to upload or download something:
location: ftp.broadinstitute.org
username: gsapubftp-anonymous
password: <blank>
location: ftp.broadinstitute.org
username: gsapubftp
password: 5WvQWSfi
If you use your browser as FTP client, make sure to include the login information in the address, otherwise you will access the general Broad Institute FTP instead of our team FTP. This should work as a direct link (for downloading only):
Geraldine Van der Auwera, PhD
Questions and comments up to August 2014 have been moved to an archival thread here:
Geraldine Van der Auwera, PhD
Hi there. Are you planning to upgrade the Broad Bundle to work with the GRCh38 genome and if so, do you know when the datasets will be available.
Best wishes,
Chris Penkett
Hi Chris,
Have a look at this thread which may answer your questions: http://gatkforums.broadinstitute.org/discussion/3843/plans-to-update-the-gatk-bundle#latest
I wasn't able to login ftp://email@example.com/bundle. It always gave me login incorrect error message. Please help. Thanks.
@yqye This looks like a temporary issue with the FTP server. I'll check with the server admin. Meanwhile, try connecting again later. Sorry for the inconvenience.
Geraldine Van der Auwera, PhD
works, is there a way to navigate to the bundle from there?
@naymikm I don't understand your question. Do you have a problem accessing the bundle contents?
Geraldine Van der Auwera, PhD
Sorry the formatting turned out weird.. I can't access the ftp server with the login credentials. user:anonymous pass: blank> works. Can I access the bundle through the ftp from there?
No, that will take you to the general Broad FTP server, not our group's. The link above worked for me when I tested it just now, does that not work for you?
Geraldine Van der Auwera, PhD
Hey it works now! It wasn't before... Thanks!
Hello, I am trying to follow this link ftp://email@example.com/bundle but cannot access the bundle. Getting the following error message: "This webpage is not available".
If the direct link is not working for you, please try using an FTP client (such as FileZilla).
Geraldine Van der Auwera, PhD


text:
Motta picked by Falcons in 7th round of NFL Draft
Thompson, Pryor signed as free agents
VERO BEACH — As the NFL Draft kept creeping closer to the finish, the anxiety kept rising for Zeke Motta.
But once Atlanta Falcons general manager Thomas Dimitroff called to let Motta know the team was going to select him, those feelings turned to pure joy.
Motta, a Vero Beach High School graduate and Notre Dame strong safety, was taken by the Falcons with the 38th pick of the seventh round of the draft on Saturday. Motta was the lone Treasure Coast player selected in the draft.
Lonnie Pryor, an Okeechobee graduate and Florida State fullback, signed with the Jacksonville Jaguars as an undrafted free agent Saturday night. Jason Thompson, a South Fork graduate and New Haven wide receiver, signed with the Washington Redskins.
"There are no words to describe how amazing this feels," Motta said. "It's awesome to know you've gotten drafted to the NFL — it's a big feat, that's for sure. Despite the round, it didn't matter. I'm so thankful and blessed to have this opportunity.
"We had plenty of feelings with teams coming and going. The anxiety buildup was to the max. (Seeing Atlanta had three picks left in the round), we thought maybe they would be the one last hope and that was them. It's a dream come true."
Motta was an honorable mention All-American as a senior at Notre Dame, but slipped in the draft due to concerns over his speed. The Falcons and Carolina Panthers were two of the teams to show the most interest in Motta prior to the draft.
Motta said he watched the entire draft and just waited to finally hear his name called.
During the seventh round of the draft, teams started calling Motta to see if he would be willing to sign with them as a free agent, if he went undrafted. He was on the phone with the Panthers when he saw a call from Georgia coming in.
"It was the GM and he was asking me if I was ready to be a Falcon. I was like, 'Are you kidding me? Of course I was!' It was a long day, but I am really enjoying it right now," Motta said. "It's so exciting. I'm going to be a 'Dirty Bird' in Atlanta."
Thompson played just one year of football at South Fork and never got a chance to play Division I football, but he will get his shot at the NFL.
Thompson said the Redskins' coaching staff was adamant about his chances of making the team and he liked the thought of playing with Washington quarterback Robert Griffin III.
"I was hoping maybe (I would be picked in) the sixth or seventh round, but knowing I was from a D-II school, I was just hoping for the best," Thompson said. "I wanted to get a foot in the door and I'm pretty happy right now."
Pryor, who also can play running back, also went undrafted, despite being one of the top fullbacks available in the draft and running the fastest 40-yard dash time of any fullback at the NFL Scouting Combine.
Pryor announced he was signing with Jacksonville on Twitter. He did not return calls seeking comment.
"Heading to the Jacksonville Jaguars as a Free Agent," Pryor tweeted. "Thankful for this opportunity and just getting started."
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text: Perfect reception, perfect location, helpful staff and also polite and nice to me and my family! Perfect room service when my parents couldn't leave the room because we spent the whole day walking in the city! They gave us a day card to walk for free with public transportation!!! Helped us how to get to places we wanted to visit,... More


text:
With high demand for office space in San Francisco these days — thanks largely to the latest technology bubble [1], Mayor Ed Lee's economic development focus [2], and its amplification by the San Francisco Chronicle [3] — Hearst Corp., which owns both the paper and the Chronicle Building, seems to be more focused on property management than journalism these days.
Following up on blogs that broke the story, Chronicle Technology Columnist James Temple today reported [4] that Yahoo is negotiating with Hearst to move its headquarters into the Chronicle Building at 5th and Mission streets. What Temple didn't say -- and what sources at the Chronicle confirmed to the Guardian, despite the fact that it hasn't yet been announced to Chronicle staff -- is that the third floor newsroom will soon be relocated while the space undergoes a renovation.
It's not clear whether the two pieces of news are related, and we're still waiting for a response to our questions on the subject from Chronicle Editor Ward Bushee. But it certainly seems true that Hearst and the Chronicle are doing everything they can to profit from the commercial real estate market that they have helped to heat up while operating a newspaper that has struggled to become profitable in recent years [5].
Valued at more than $30 million and covering nearly a full city block in the heart of the city, the Chronicle Building has been steadily taken over by outside companies in recent years, many of them technology corporations such as Square, the online payment company. The newsroom that used to occupy the second and third floors has already been squeezed onto the third, and now even that space is getting an overhaul.
Meanwhile, Hearst has been working with Forest City [6] and Strada Investment Group on a plan to redevelop the property [7], reportedly replacing the old Hearst [8] headquarters and other buildings that share the block with an office and residential tower and trying to win historic landmark status [9] for the Chronicle Building itself.
Chronicle staffers tell the Guardian that they were surprised to hear about the newsroom relocation last week and they don't have many details, except that they will remain in the building. And given how valuable it has become, they say they're just happy to not be totally squeezed out by the tech boom.
[1] http://www.sfbg.com/2012/02/14/bubble-back
[2] http://www.sfbg.com/politics/2013/04/19/ten-more-ron-conways-would-destroy-san-francisco
[3] http://www.sfbg.com/politics/2011/05/19/chronicle-pushes-fake-campaign-draft-ed-lee
[4] http://www.sfchronicle.com/technology/article/Yahoo-zeroing-in-on-space-at-Chronicle-Building-4694287.php?t=8055fa6036a85878bf
[5] http://www.sfbg.com/2009/03/04/chronicle-death-watch?page=0,0
[6] http://www.sfgate.com/news/article/Hearst-plans-major-development-around-Chronicle-2289133.php
[7] http://www.sfbg.com/politics/2011/11/18/chron-owner-exploits-its-most-valuable-asset
[8] https://www.baycitizen.org/news/business/chronicle-buildings-transformation/
[9] http://www.sfbg.com/politics/2011/02/07/richard-johns-closer-developers-preservationists


text:
The location and orientation of a barcode block is determined by first producing an image of the document that excludes the barcode block and producing a second image wherein the spaces in the barcode block have been deleted thereby providing a solid rectangular block. The two images are subtracted to reject large background clutter while displaying with certainty the full image of the barcode block. Thereafter, down-sampling occurs to remove small clutter and other non-barcode objects while locating the centroid of each barcode block. The image containing the centroid is up-scaled and used to calculate the position of the barcode block in the original document for use by a barcode reader.
1. A method of determining the location of a barcode block having a predetermined size and shape on a document having extraneous printed matter thereon comprising:
receiving a digital image of a scanned document as an input signal;
reducing the input signal to produce a first down-sampled image that is over-sampled;
reducing the input signal to produce a second down-sampled image that is under-sampled;
deleting barcode image pixels from the under-sampled image to produce a barcode free image;
subtracting the barcode free image from the over-sampled image to produce an image definitely showing the barcode block location but not showing large sized background clutter that is common to the barcode free image and the over-sampled image; and
down-sampling the barcode block location containing image and thereafter removing small sized background clutter thereby to produce an image showing the barcode block location which is substantially free of background clutter.
2. The method of claim 1 wherein the over-sampled and undersampled images are respectively produced by:
forming one sub-pixel from four pixels from the original image with said one sub-pixel being a 1 if any of the said four original image pixels are a 1; and
forming one sub-pixel from four pixels from the original image with said one sub-pixel being a 1 only if all of said four original pixels are a 1.
3. The method of claim 1 further including the step of modifying the over-sampled image to connect all bars in a barcode into a single rectangular area prior to the subtracting step.
4. A method of determining the location of all barcode blocks having a predetermined size and shape on a background area that is scanned to produce an image of the area in a data format comprising the steps of:
reducing input image data into two down-sampled images, one being under-sampled and the other being over-sampled;
modifying the two down-sampled images so that an image of each barcode block is positively provided in a third image and positively not present in a fourth image;
subtracting the fourth image from the third image to produce a fifth image that contains the barcode block and unwanted small clutter but is free of clutter present in both the third and fourth images; and
thereafter removing the unwanted small clutter from the fifth image to provide a sixth image containing only the barcode block.
5. A method of claim 4 wherein the removal of unwanted small clutter from the fifth image includes the step of providing a disk having a radius of a predetermined number of pixels that is effective to eliminate images of objects forming small clutter that are smaller than the barcode block.
6. A method of detecting the location of a barcode having a predetermined size and polygonal shape on a background containing printed matter for subsequent reading of the barcode comprising:
reducing by a first reduction factor input image data of the background and barcode to produce a first over-sampled image and a second under-sampled image;
removing all pixels that form the barcode by morphological erosion from the under-sampled image to produce a modified under-sampled image;
connecting all bars in a barcode into a single area having said polygonal shape by morphological dilation from the over-sampled image to produce a modified over-sampled image; and
subtracting said modified under-sampled image from the modified over-sampled image to produce a new image that contains the barcode shape at its position on the background, said new image being free of large size clutter that is part of both the under-sampled image and the over-sampled image.
7. The method of claim 6 wherein the morphological erosion comprises use of a 5�5 pixel structuring element to remove all pixels that may have been part of a barcode and the morphological dilation comprises use of a 5�5 pixel structuring element that is effective to connect all bars in a barcode into a single rectangular area.
8. The method of claim 7 comprising the further steps of:
reducing by a second reduction factor said new barcode containing image to reduce the processing time for subsequent steps including removal of small, size clutter and analyzing data involving connected pixels of an object.
9. The method of claim 8 wherein the connected pixel object analysis comprises:
detecting sets of pixels in said new barcode containing image that are all connected by a path of pixels that are within one pixel of each other;
determining the direction and location in said image of a centroid of each object including said barcode image;
rejecting objects that do not have a barcode shape; and
up-scaling the new barcode containing image to provide location of the centroid in a full resolution image and enable calculation of the position of the corners of the full resolution barcode image.
10. A method of reducing clutter from a document image containing printed matter including at least one barcode having dark bars and blank spaces in a configuration having a predetermined shape and area comprising:
generating a pixel based image of said at least one barcode in a data generated pixel based image of the document in which printed matter appears as objects and all bars of the barcode appear as an object comprised of a solid bar without spaces and having said predetermined shape;
forming a table of said objects in said pixel based image including at least one barcode and clutter, each of said objects having an area with the object area being defined by the number of pixels in the area; and
eliminating from the pixel based image all objects which have a number of pixels that differs by more than a predetermined amount from the number of pixels in the area of a genuine barcode image.
11. The method of claim 10 further comprising eliminating from the pixel based image all remaining objects having a linear dimension which exceeds a dimension of a genuine barcode image.
12. The method of claim 10 wherein the pixel based image containing a barcode in which all bars appear as a solid bar without spaces is generated by:
reducing input image data from an original document into two down-sampled images, one being under-sampled and the other being over-sampled;
modifying the two down-sampled images so that an image of the barcode block is positively provided in a third image and positively not present in a fourth image;
subtracting the fourth image from the third image to produce a fifth image that contains the barcode block and unwanted small clutter but is free of clutter present in both the third and fourth images; and
thereafter removing the unwanted small clutter from the fifth image to provide a sixth image including the barcode and objects not removed as large clutter or small clutter.
13. A method of claim 12 wherein the removal of unwanted small clutter from the fifth image includes the step of utilizing a disk having a radius of a predetermined number of pixels that is effective to eliminate images of objects that are smaller than the barcode block.
This invention relates to barcode detection systems, and more particularly, to a method of accurately determining the location of one or more barcodes on a document.
When a barcode symbol is included within a document containing other information, there are two problems. The first is how to identify the barcode location. The second problem relates to reading the thus located barcode data. The present invention addresses only the first problem, and is mainly concerned with how to distinguish the barcode from other markings such as printed matter on the document.
One prior technique for locating and reading a barcode is disclosed in Sarna et al, U.S. Pat. No. 4,873,426 where scanning is in a direction generally perpendicular to the orientation of the code bars. Such scanning is on a row-by-row basis to identify rows above and below an identified row that matches certain stored parameters. The row most likely to contain a barcode is read. Only a single barcode can be identified on a document page.
Another prior technique for determining barcode location is disclosed in commonly owned co-pending Lee et al, U.S. Patent Application Ser. No. 07/289,662 filed Dec. 22, 1988, now U.S. Pat. No. 4,948,955. The techniques there disclosed locate the barcode at any position or direction using an initial down-sampling to a coarse resolution thereby to reduce the time required for location determination at a sacrifice of image resolution.
It is an object of the present invention to provide a novel process which has improved resolution and can be executed in a short time period to provide an improved capability of locating one or more barcodes on a background. The method involves modifying any barcode image to eliminate all spaces between the bars and removing large clutter in an image that has been down-sampled by a relatively small amount. Thereafter, the image is further downsampled to speed up the process of small clutter removal and the determination of a centroid location of each barcode image. The centroid data is up-scaled and used to identify all actual barcode locations for use in a barcode reader.
In producing a barcode image free of spaces, the present invention uses two particularly down-sampled images, one being over-sampled and the other being under-sampled. Both images are modified so that in the under-sampled image, all pixels of the barcode are deleted whereas in the modification to the over sampled image, all spaces in the barcode are deleted to form a solid rectangle. Subtracting the two images to delete all common areas reliably leaves a solid barcode image while removing large clutter. Small clutter that remains and further clean-up can be quickly accomplished after the second down-sampling.
The foregoing features and other objects of the invention will become more fully apparent from the claims and from the description when read in conjunction with the appended drawings.
FIG. 1 is a view of a document having a randomly positioned barcode;
FIG. 2 is a block diagram of a system for determining the location of a barcode block;
FIG. 3 is a view of a down-sized image that has been undersampled;
FIG. 4 is a view of a down-sized image which has been oversampled;
FIG. 5 is a view of the image of FIG. 3 that has been modified to remove 11 pixels representing the barcode;
FIG. 6 is a view of the image of FIG. 4 that has been modified to remove all spaces in the barcode so that the barcode appears as a solid rectangle;
FIG. 7 is a view resulting from the subtraction of the FIG. 5 image from the FIG. 6 image thereby to show the solid barcode rectangle its actual position on the document; and
FIG. 8 is a view where the small clutter has been removed.
The invention will be described in connection with a document that may be a page 10 as illustrated in FIG. 1 having a size of 8.5"�14" and a barcode 12. However, the invention is applicable to locating the position of each barcode, in the event more than one barcode is present, on a background of any size that can be viewed with appropriate equipment that converts a visible image into a data format using pixels.
The barcode 12 consists of bars separated by spaces and represents a block of uniform area within a known shape, usually a polygonal shape that is customarily rectangular. The location of the barcode block 12 on a document 10 is variable and its major axis may have a random orientation with respect to marginal edges of the document 10.
The document 10 usually has printed matter which may include logos and other blocks of printing that are at least as large as the barcode block 12 which is referred to herein as a large clutter. Other areas of printed matter may include small print in lines of thin texts, border lines or signatures, which is referred to herein as small clutter. These documents are commonly used, for example, in the transport industry and receiving stations have barcode readers that are used for converting the barcode data into appropriate signals that are used in a data processing system.
One problem which introduces delay into the processing of these documents relates to finding the barcode 12 that is on the document. Scanners conventionally have a resolution of 200 pixels per inch. The barcode 12 may be in any location or orientation on the document and the background content of the document is arbitrary.
No feature of the barcode block, which frequently is on a label having an adhesive on its back surface, may be used to determine its location other than its size and shape of the rectangular area containing the bars and spaces. The location accuracy must be adequate to allow barcode reading by a suitable barcode reader such as that disclosed in a commonly owned copending application Ser. No. 07/289,663 filed Dec. 22, 1988, now abandoned.
A first step in the process according to the invention is to produce an image of the block containing the barcode at its location on the document. This image should be free of background noise that would generate false readings from the barcode reader.
A preferred procedure according to one feature of the present invention involves producing down-sampled images of the document by reducing the number of pixels from 200 per inch to, for example, 50 new pixels per inch which would be a reduction by a factor of four. Each new pixel is referred to as a sub-pixel.
In FIG. 2, this reduction or down-sizing is illustrated as accruing in a first stage 20 where the image is under-sampled. Sub-pixels in the output image are equal to one only if all four of the pixels in the input image signal on line 24 are one. FIG. 3 illustrates a down-sized image which is under-sampled and is available on line 24. It should be noted that barcode lines are of reduced clarity.
At a second stage 26 which is in parallel with the first stage 20, the down-sized image is over-sampled. Sub-pixels in the output image are equal to one if any of the four pixels in the input image signal on line 22 is one. FIG. 4 illustrates a down-sized image over-sampled and is available on line 28. Here spaces in the barcode are partially lost.
To reliably guarantee that each barcode block that is on the document is preserved, the under-sampled image is first subjected to a process such as morphological erosion in block 30 of FIG. 2 such that each pixel in the barcode of FIG. 3 is rejected. This means that a barcode free image is produced as illustrated in FIG. 5. Large clutter objects 32 and 34 remain which is advantageous. To effect this morphological erosion, a 5�5 pixel structuring element may be done on the under-sampled FIG. 3 image to remove all pixels that may have been part of the barcode 12.
The next step at stage 36 of FIG. 2 is to operate on the FIG. 4 image to remove all spaces in the barcode image so that the barcode is presented as a solid rectangle as shown in FIG. 6. Clearly, the large clutter objects remain in the FIG. 6 image and small clutter 38 from the FIG. 4 image is retained. The small clutter 38 is not retained in FIG. 5. To effect the morphological dilation, a 5�5 pixel structuring element may be used.
Thereafter, the image of FIG. 7 is obtained by subtracting the FIG. 5 image from the FIG. 6 image as indicated in block 40 of FIG. 2. The large clutter objects 32 and 34 are rejected. The barcode block 12 of FIG. 6 is retained along with the small clutter regions 38 of FIG. 4 that were connected in the dilation step which form the image of FIG. 6 as indicated by block 42 of FIG. 2.
An advantageous feature of the present invention is that at this point a second image down-sampling to one quarter size may be effected by image data reduction stage 44 so that a document that is 1/16th of the original document size can be used. This reduces the time required to execute succeeding morphological operations since the execution time is linearly proportional to image size. However, down-sizing to 1/16th of the original document size before obtaining a solid barcode block image has been found to give unreliable results.
Small clutter rejection as indicated by block 46 is next carried out as by use of a morphological opening with a disk of radius 9. Such clutter objects 38 are too small to be barcodes.
Clutter objects sometimes remain and additional steps that can be taken include making an area test, a line-run length test and an extreme boundary test. In addition, the centroid of a barcode image is needed to identify its orientation and aid in calculating the extreme corner positions.
At stage 48 of FIG. 2, the connected areas of each remaining object image are analyzed to determine the centroid, an area, a bounding box, a major axis moment, a minor axis moment, and an angle with respect to the horizontal axis. This information is used to distinguish real barcode rectangles from clutter objects. The expected values and tolerances for the following test are determined empirically.
Run-length encoding involves connected components which are sets of pixels in the image that are all connected by a path of pixels that are within one pixel of each other. A table of objects is generated by scanning the image one line at a time from the first to the last line in the image. For each line, the algorithm locates pixels connected in a line (run-lengths). The run-lengths for a line are compared to the run-lengths for the previous line to up-date object information. The run-data (start pixel and length) are used to calculate geometrical properties of the object.
An area test eliminates objects from the image that do not fall within a small percentage of the expected area of a genuine barcode image. The area of an object is equal to the number of pixels in the connected area of the object. This value is determined by adding the lengths of all runs that are part of the object and deviations from the predetermined number in the barcode block indicate clutter. This test typically eliminates objects resulting from a large area of small clutter in the document such as a paragraph of very fine print.
The angle between a major axis of each remaining object and the horizontal axis is calculated from the second moment of the object about its major axis. Using this angle and the dimensions of the barcode, a calculation is made of the extreme x and Y values a valid barcode object should attain. Testing the extreme values of an object will eliminate objects that have the correct area but the wrong shape. At this point in the process, few if any clutter objects remain.
A shape disk may be used which compares the ratio of moments about the major and minor axis within accepted limits based on the rectangular shape of the barcode. This is an even more restrictive shape test in that it is sensitive to objects that are not rectangles, but which otherwise might pass all previous tests.
Finally, the centroid of the barcode that has earlier been found during the connected component analyses is up-scaled by 16 as indicated at image enlargement stage 50 of FIG. 2. By this step, the centroid is obtained in the full resolution image. The centroid and the angle of its major axis with respect to the horizontal axis are then used to calculate the position of the four corners of the full resolution barcode image. These parameters are passed to a barcode reading program such as that described in application Ser. No. 07/289,663 identified above.
In summary, the present invention may be characterized by providing a barcode image in which all spaces are obliterated to provide a solid mark in contrast with the background. This mark is located and oriented in its actual position. Larger clutter is rejected when the solid mark is formed. Thereafter, a further down-sizing can be used to reduce data processing time and small clutter rejected thereby leaving objects that are about the size of the barcode. Object area comparisons with the known barcode size allows removal of some additional clutter and the step of determining centroids of the remaining objects allows rejection of improperly shaped objects and identification of the orientation and corner points of the barcode image which is up-sized to its original image size.
While the present invention has been described with reference to a particular system, other variations and modifications will occur to those skilled in the art. Such variations and modifications as fall within the scope of the claims or equivalents thereof are intended to be covered thereby.
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2.6 miles | 3804 Peachtree Ave., Ste. 110, Wilmington, NC 28403
Attorney at Law
0.7 miles | 310 North Front Street, Suite 200, Wilmington, NC 28401
Our attorneys practice in a variety of areas of the law. We strive to exceed your expectations for legal services in an more...
Our attorneys practice in a variety of areas of the law. We strive to exceed your expectations for legal services in an ethical, expeditious and cost effective manner. less...
0.4 miles | 401 Chestnut Street, Suite G, Wilmington, NC 28401
Rice Law focuses on what matters
Serving Wilmington, NC 28401
A highly rated Law Firm practicing Child Support law.
Serving New Hanover County, North Carolina
0.3 miles | 406 Market Street, Wilmington, NC 28402
A highly rated Law Firm established in 1983 practicing Child Support law. Accepts credit cards.
Child support is a court-ordered payment a non-custodial parent must make to the custodial parent to cover part of the expenses of raising the child. This includes housing and utilities, food, clothing, education expenses, and other costs. Find child support legal articles and contact a Wilmington, North Carolina child support law firm from the list of results on this page provided by Lawyers.com.


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A scary ghost creature in the Mushroom Kingdom. Mario faces these a lot. These pearly-white ghosts will become invisible if anyone faces them. They usually cannot be disposed of. There are also other Boos, like the Big Boo, King Boo, and more.
"I thought I heard a Boo, but I turned around and saw nothing."
ayon kay SeaCaptainDrake ika-06 ng Disyembre, 2003
This word is used in a replacement of any cuss word.
Id boo that !!!
My left ankle hurts like a freaking boo !!!
Ill will boo you up if you dont shut up !!!
My left ankle hurts like a freaking boo !!!
Ill will boo you up if you dont shut up !!!
ayon kay londonengland112 ika-07 ng Pebrero, 2010
Basically it means like forget you.
*1. Someone keeps staring at you, you say "Boo Bitch! Whatchu lokkin at?!?!?!"
*2. Girl 1: Hey can you hang out today???
Girl 2: No sorry...
Girl 1: Boo, you whore!!! You suck.
*2. Girl 1: Hey can you hang out today???
Girl 2: No sorry...
Girl 1: Boo, you whore!!! You suck.
ayon kay lol.it.babe=) ika-03 ng Hunyo, 2009
An ancient mating ritual involving the death of the male.
Bob: Come Charlie! Lets do some boo.
Charlie: I'll see you on the flipside...
Charlie: I'll see you on the flipside...
ayon kay Oilhater ika-14 ng Hunyo, 2008
a term used in prison referring to someone as your bitch but the term has been taken out by the black man to the urban streets as your loved one
black man- "yea thats right mother fucker you take it cuz you always gonna be my boo"
ayon kay nick zanko ika-15 ng Agosto, 2006
The name of the Peterson's pet cat (until dad let it go stray right round near the animal auction).
Oh no, Boo peed on the red couches again!
ayon kay Danny Peterson ika-26 ng Enero, 2004
something you say when someone is stareing right at you and you look at them and they are still looking at you, you say boo in a wat are you looking at voice and they stop looking at you!!!
girl#1:(looks at a gurl and she looking at her) boo wat you looking at lil ugly
girl#2:rolls her eyes and stops looking
girl#2:rolls her eyes and stops looking
ayon kay *gloria* ika-22 ng Abril, 2008


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Adobe Photoshop Lightroom 4 hits the Mac App Store for $150
Along with the $80 Photoshop Elements, Premiere Elements and Revel, Adobe now offers its popular pro/sumer photo management software Lightroom 4 in the Mac App Store for $150. Though it generally gets better reviews than Apple's own $80 Aperture, it is almost double the price and can often be found offline for less. Adobe fans […]
Adobe touts half-million Photoshop CS6 Beta downloads, previews Illustrator CS6
On the Photoshop Blog, Adobe today said it passed 500,000 downloads with its free Photoshop CS6 Beta software. As soon as it was announced, Photoshop CS6 beta was a worldwide Trending Topic Our sneak peek video playlist on YouTube has received nearly 3 million views The Photoshop CS6 beta videos have already received half a million views We rounded up the reviews […]
Photoshop CS6 Review roundup and free beta download
We reported this week that Adobe was "wrapping up CS6 development and that a public beta was launching soon". The review embargo has just lifted and the beta is out. Here are some reviews from around the Web: Time's Harry McCracken says it is a little magical. Bjango: Improvements! Gizmodo: Best Release in recent history CNet's […]
Adobe announces Photoshop Lightroom 4 availability, upgrades from $79
Photoshop maker Adobe, Inc. today announced in a blog post general availability of its Aperture competitor Lightroom. A free beta of Photoshop Lightroom 4 was posted two months ago and today the software is available for education customers on both Mac OS X and Windows platforms. The company's Vice President of Products and Creative Media […]
Adobe Photoshop Touch hits iPad 2 for US$9.99 (Update: Back up!)
Note – Photoshop Touch just started hitting AU/NZ. Update: It was pulled, but it is back ($9.99 at iTunes) Photoshop maker Adobe Systems, Inc., released a long-expected iPad companion aptly named "Photoshop Touch." The Android version demonstrated at Maxx earlier this year and released shortly afterwards. The first in a series of six touch-optimized apps (the other […]
Adobe demos content-aware move in Photoshop CS6
Software-maker Adobe posted an interesting video on its Photoshop channel through YouTube. The clip showcases content-aware move, which is one of the new features in the upcoming Photoshop CS6 update. Similar to content-aware fill in Photoshop CS5 that replaces the missing pixels behind removed image elements, the new feature lets you define a region and […]
Adobe details Creative Cloud: $49.99 membership fee, 20GB storage, CS6, Lightroom 4, Muse, Edge, Photoshop touch apps
Adobe just revealed (via The Verge) pricing information and more details on its new cloud-based offering called Creative Cloud. Announced last October along new touch apps for tablets, Creative Cloud is billed as a service that "radically redefines the content creation process" by offering a central web-based repository to store, share and view content across devices. Prices start […]
Apple updates Final Cut Pro X with multicam editing, broadcast monitoring, Photoshop support and more (Motion and Compressor, too)
Update: Apple has published a white paper entitled "Final Cut Pro X for Final Cut Pro 7 Editors" to detail the differences between the two apps and smooth the transition for professional users. Apple updated its Final Cut Pro X video editing software this morning with some much-needed improvements. It is a significant update, because Final Cut Pro X […]
Adobe Photoshop Elements 10 and Premiere Elements 10 hit Mac App Store
Two short months after Adobe released Photoshop Elements 10 and Premiere Elements 10 in boxed form for $99/each or $149 combo, the apps have now arrived in the Mac App Store, priced at $79.99/each. Photoshop Elements 10 (App store) and Premiere Elements 10(App Store) became available overnight with the same functionality improvements that the box versions produced, including: New Facebook features allow […]
Adobe Photoshop Lightroom: $150
From the Toys Section: . Amazon offers Adobe Photoshop Lightroom 3 for Windows and Mac, model no. 65064073, for $149.99. With free shipping, that's tied with our October mention of a downloaded version and the lowest total price we could find by $130. This photo editing software is designed for use with RAW files. Adobe charges $89 to download […]
Photoshop "unblur" leaves MAX audience gasping for air
Damn impressive. I'm with the audience at 1:18 and again later when Kevin Lynch's picture is unblurred. I just hope I kept all of those amazing but flawed pictures that I can now unblur. via TNW Update: A better version is over at Adobe.
Adobe unveils Adobe Creative Cloud, Photoshop Touch and five other tablet apps
Update: We go hands on with new Adobe Touch Apps! Adobe Photoshop Touch and five other new iPad apps from Adobe will land on the App Store soon, coming to Android devices in November. Adobe today made a series of announcements at their MAX conference which runs October 1-5. The company doubled down on its […]
First demo with Photoshop Touch #MAX2011
Update: Photoshop Touch is now available for the iPad I am hands on here at Adobe Max where they are demo-ing Photoshop touch. They said the Android version was further along(!!). Adobe Collage Touch and others below:
Adobe releases Photoshop Elements 10 & Adobe Premiere Elements 10
Adobe today quietly released Photoshop Elements 10 and Premiere Elements 10, bringing with them new Facebook and YouTube integration, video editing and burring features, object-based search, and new color correcting and text curving and flowing effects. New Facebook features allow you to auto analyze your images to identify people and tag them based on your […]
Adobe makes its first foray into the Mac App Store with Photoshop Elements
Macworld reports that Adobe is making its first application available on the Mac App Store. Adobe Photoshop Elements 9 Editor is being offered for $79.99 which is less than the $99 boxed version (which is often discounted– for instance it is $60 currently after a $20 discount at Amazon). Mac App Store version of the application costs $80 and includes […]
Adobe Photoshop Lightroom 3: $120
From the 9to5toys.com: Today only, Amazon.com offers the Adobe Photoshop Lightroom 3 for Windows and Mac, model no. 65064073, for $119.99 with free shipping. That's $30 under our November mention and the lowest total price we've seen. (It's also a current price low by $60.) This software offers a variety of photo-editing tools.
Photoshop iPad apps finally hit the App Store
Adobe, which last week updated Photoshop to work with its freshly announced iPad apps, has finally released said programs for iOS devices. Adobe Eazel, Adobe Nav and Adobe Color Lava, iPad companion apps for Photoshop, are now available for download on the App Store. Adobe Eazel is a finger-paint program that sells for five bucks. The […]
Adobe updates Photoshop to work with new iPad apps Adobe Nav, Adobe Color Lava and Adobe Eazel
Adobe has just updated Photoshop to version 12.0.4 this morning. It includes the promised support for Adobe's new iPad apps designed to work with desktop Photoshop: Adobe Nav, Adobe Color Lava and Adobe Eazel. Said apps are in the process of begin added to the App Store and are not live yet on all regional App […]
Adobe Photoshop Elements 9: $40
From the Toys Section: Today only, Amazon.com offers Adobe Photoshop Elements 9 for Windows and Mac, model no. 65089532, for $59.99. The $20 mail-in rebate drops it to $39.99. With free shipping, that's tied with last month's mention, the lowest total price we could find by $20, and within a buck of the best deal we've ever seen on […]
Tidbits: Adobe TV now iOS-friendly, Photoshop Remote for iPad demoed
Hot off the presses, new tidbits hit us following the launch of Adobe Creative Suite 5.5 and new iPad apps from Adobe, both announced this morning. TUAW reports that Adobe's video training resources now support iOS-friendly streaming to iPhones, iPods and iPads. Creative pros can now learn new tricks as they commute on train, go […]


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HOPKINTON, Mass., Nov. 21 /PRNewswire-FirstCall/ -- Alseres Pharmaceuticals, Inc. (Nasdaq: ALSE), today announced that it has entered into a securities purchase agreement with Robert Gipson for the private placement of 543,478 shares of common stock of the Company, together with warrants to purchase 543,478 additional shares of common stock at an exercise price of $1.84 per share, for a total purchase price of $1.84 per share. The financing resulted in total gross proceeds of $1 million. The Company intends to use the net proceeds from this private placement to fund its research and development programs and otherwise for general corporate purposes.
(Logo: http://www.newscom.com/cgi-bin/prnh/20080813/NEW056LOGO )
The securities offered and sold by Alseres in this private placement have not been registered under the Securities Act of 1933, as amended (the "Securities Act"), or state securities laws and may not be offered or sold in the United States absent registration with the Securities and Exchange Commission ("SEC") or an applicable exemption from registration requirements. Alseres has agreed to file a registration statement with the SEC covering resale of the shares of common stock and the shares of common stock issuable upon exercise of the warrants issued in the private placement.
This notice is issued pursuant to Rule 135c under the Securities Act and shall not constitute an offer to sell or the solicitation of an offer to buy, nor shall there be any sale of these securities in any state in which such offer, solicitation or sale would be unlawful prior to the registration or qualification under the securities laws of any such state.
About Alseres Pharmaceuticals, Inc.
Alseres Pharmaceutical
|SOURCE Alseres Pharmaceuticals, Inc.
Copyright©2008 PR Newswire.
All rights reserved


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Browse blinds and canopies in Lincolnshire featuring photos, videos, vouchers and blinds and canopies reviews to help you choose the right local blinds and canopies for you.
Why not list your blinds and canopies business on Lincolnshire Echo?
Address: 9b Laughton Road, Blyton, Gainsborough, DN21 3LG
Address: , , BOSTON, PE20 3QF
Address: The Old School Station Road East, , GRANTHAM, NG31 6HX
Address: Unit 3 Edlington Court, , GAINSBOROUGH, DN21 1LT
Address: Nettleham House, East Street, LINCOLN, LN2 2SL
Address: 13 Collingham Road, , Lincoln, LN6 9LP
Address: 14 Royds Crescent, , WORKSOP, S80 3HF
Address: 12 Cleveland Avenue, North Hykeham, Lincoln, LN6 9TH
Address: 52 Saddlers Way, Fishtoft, Boston, PE21 0BB


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As he recounts and interprets the lessons of the Gospel, Girzone shows that compassion for our fellow human beings is the most profound expression of God's love and wisdom. His thoughtful, incisive meditations will help readers turn away from thoughts of retribution or punishment, and will inspire them to embrace God's message of help and healing.
©2009 Joseph F. Girzone; (P)2009 Brilliance Audio, Inc.
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Set of 100 paper linen Caspari napkins. These air-laid paper napkins have the look of linen with a thicker, plusher, more luxe feel. Available in beverage/cocktail size or luncheon size. You select imprint color, lettering style and positioning for your personalization.
Caspari is committed to producing products using environmentally sound raw materials. Beautifully packaged and ready for gift giving. Font size is determined by the typesetter depending on length of copy.