A compound of the formula (I) is disclosed which is a vitronectin receptor antagonist and is useful in the treatment of osteoporosis:

This invention relates to a pharmaceutically active compound which inhibits the vitronectin receptor and is useful for the treatment of inflammation, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and diseases wherein bone resorption is a factor, such as osteoporosis.
Integrins are a superfamily of cell adhesion receptors, which are transmembrane glycoproteins expressed on a variety of cells. These cell surface adhesion receptors include gpIIb/IIIa (the fibrinogen receptor) and xcex1vxcex23 (the vitronectin receptor). The fibrinogen receptor gpIIb/IIIa is expressed on the platelet surface, and mediates platelet aggregation and the formation of a hemostatic clot at the site of a bleeding wound. Philips, et al., Blood., 1988, 71, 831. The vitronectin receptor xcex1vxcex23 is expressed on a number of cells, including endothelial, smooth muscle, osteoclast, and tumor cells, and, thus, it has a variety of functions. The xcex1vxcex23 receptor expressed on the membrane of osteoclast cells mediates the adhesion of osteoclasts to the bone matrix, a key step in the bone resorption process. Ross, et al., J. Biol. Chem., 1987, 262, 7703. A disease characterized by excessive bone resorption is osteoporosis. The xcex1vxcex23 receptor expressed on human aortic smooth muscle cells mediates their migration into neointima, a process which can lead to restenosis after percutaneous coronary angioplasty. Brown, et al., Cardiovascular Res., 1994, 28, 1815. Additionally, Brooks, et al., Cell, 1994, 79, 1157 has shown that an xcex1vxcex23 antagonist is able to promote tumor regression by inducing apoptosis of angiogenic blood vessels. Thus, agents that block the vitronectin receptor would be useful in treating diseases, such as osteoporosis, restenosis and cancer.
The vitronectin receptor is now known to refer to three different integrins, designated xcex1vxcex21, xcex1vxcex23 and xcex1vxcex25. Horton, et al., Int. J. Exp. Pathol., 1990, 71, 741. xcex1vxcex23 binds fibronectin and vitronectin. xcex1vxcex23 binds a large variety of ligands, including fibrin, fibrinogen, laminin, thrombospondin, vitronectin, von Willebrand""s factor, osteopontin and bone sialoprotein I. xcex1vxcex25 binds vitronectin. The vitronectin receptor xcex1vxcex25 has been shown to be involved in cell adhesion of a variety of cell types, including microvascular endothelial cells, (Davis, et al., J. Cell. Biol., 1993, 51, 206), and its role in angiogenesis has been confirmed. Brooks, et al., Science, 1994, 264, 569. This integrin is expressed on blood vessels in human wound granulation tissue, but not in normal skin.
The vitronectin receptor is known to bind to bone matrix proteins which contain the tri-peptide Arg-Gly-Asp (or RGD) motif. Thus, Horton, et al., Exp. Cell Res. 1991, 195, 368, disclose that RGD-containing peptides and an anti-vitronectin receptor antibody (23C6) inhibit dentine resorption and cell spreading by osteoclasts. In addition, Sato, et al., J. Cell Biol. 1990, 111, 1713 discloses that echistatin, a snake venom peptide which contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits attachment of osteoclasts to bone.
It has now been discovered that a certain compound is a potent inhibitor of the xcex1vxcex23 and xcex1vxcex25 receptors. In particular, it has been discovered that such compound is more potent inhibitors of the vitronectin receptor than the fibrinogen receptor.
This invention comprises a compound of the formula (I) as described hereinafter, which has pharmacological activity for the inhibition of the vitronection receptor and is useful in the treatment of inflammation, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and diseases wherein bone resorption is a factor, such as osteoporosis.
This invention is also a pharmaceutical composition comprising a compound according to formula (I) and a pharmaceutically carrier.
This invention is also a method of treating diseases which are mediated by the vitronectin receptor. In a particular aspect, the compound of this invention is useful for treating atherosclerosis, restenosis, inflammation, cancer and diseases wherein bone resorption is a factor, such as osteoporosis.
This invention comprises a novel compound which is a more potent inhibitor of the vitronectin receptor than the fibrinogen receptor. The novel compound comprises a dibenzocycloheptene core in which a nitrogen-containing substituent is present on one of the aromatic six-membered rings of the dibenzocycloheptene and an aliphatic substituent containing an acidic moiety is present on the seven-membered ring of the dibenzocycloheptene. The dibenzocycloheptene ring system is believed to orient the substituent sidechains on the six and seven membered rings so that they may interact favorably with the vitronectin receptor. It is preferred that about twelve to fourteen intervening covalent bonds via the shortest intramolecular path will exist between the acidic group on the aliphatic substituent of the seven-membered ring of the dibenzocycloheptene and the nitrogen of the nitrogen-containing substituent on one of the aromatic six-membered ring of the dibenzocycloheptene.
This invention comprises a compound of formula (I):
or a pharmaceutically acceptable salt thereof.
The compound of formula (I) inhibits the binding of vitronectin and other RGD-containing peptides to the vitronectin receptor. Inhibition of the vitronectin receptor on osteoclasts inhibits osteoclastic bone resorption and is useful in the treatment of diseases wherein bone resorption is associated with pathology, such as osteoporosis and osteoarthritis.
In another aspect, this invention is a method for stimulating bone formation which comprises administering a compound of formula (I) which causes an increase in osteocalcin release. Increased bone production is a clear benefit in disease states wherein there is a deficiency of mineralized bone mass or remodeling of bone is desired, such as fracture healing and the prevention of bone fractures. Diseases and metabolic disorders which result in loss of bone structure would also benefit from such treatment. For instance, hyperparathyroidism, Paget""s disease, hypercalcemia of malignancy, osteolytic lesions produced by bone metastasis, bone loss due to immobilization or sex hormone deficiency, Behcet""s disease, osteomalacia, hyperostosis and osteopetrosis, could benefit from administering a compound of this invention.
Additionally, since the compound of the instant invention inhibits vitronectin receptors on a number of different types of cells, said compound would be useful in the treatment of inflammatory disorders, such as rheumatoid arthritis and psoriasis, and cardiovascular diseases, such as atherosclerosis and restenosis. The compound of Formula (I) of the present invention may be useful for the treatment or prevention of other diseases including, but not limited to, thromboembolic disorders, asthma, allergies, adult respiratory distress syndrome, graft versus host disease, organ transplant rejection, septic shock, eczema, contact dermatitis, inflammatory bowel disease, and other autoimmune diseases. The compound of the present invention may also be useful for wound healing.
The compound of the present invention is also useful for the treatment, including prevention, of angiogenic disorders. The term angiogenic disorders as used herein includes conditions involving abnormal neovascularization. Where the growth of new blood vessels is the cause of, or contributes to, the pathology associated with a disease, inhibition of angiogenisis will reduce the deleterious effects of the disease. An example of such a disease target is diabetic retinopathy. Where the growth of new blood vessels is required to support growth of a deleterious tissue, inhibition of angiogenisis will reduce the blood supply to the tissue and thereby contribute to reduction in tissue mass based on blood supply requirements. Examples include growth of tumors where neovascularization is a continual requirement in order that the tumor grow and the establishment of solid tumor metastases. Thus, the compound of the present invention inhibit tumor tissue angiogenesis, thereby preventing tumor metastasis and tumor growth.
Thus, according to the methods of the present invention, the inhibition of angiogenesis using the compound of the present invention can ameliorate the symptoms of the disease, and, in some cases, can cure the disease.
Another therapeutic target for the compound of the instant invention is eye diseases chacterized by neovascularization. Such eye diseases include corneal neovascular disorders, such as corneal transplantation, herpetic keratitis, luetic keratitis, pterygium and neovascular pannus associated with contact lens use. Additional eye diseases also include age-related macular degeneration, presumed ocular histoplasmosis, retinopathy of prematurity and neovascular glaucoma.
This invention further provides a method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I) and an antineoplastic agent, such as topotecan and cisplatin.
The novel compound of this invention is (S)-10,11-dihydro-3-[2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid or a pharmaceutically acceptable salt thereof.
According to the present invention, the (S) configuration of the formula (I) compound is preferred.
Also included in this invention are prodrugs of the compound of this invention. Prodrugs are considered to be any covalently bonded carriers which release the active parent drug according to formula (I) in vivo. Thus, in another aspect of this invention are novel prodrugs, which are also intermediates in the preparation of the formula (I) compound, of formula (II):
or a pharmaceutically acceptable salt thereof.
Abbreviations and symbols commonly used in the peptide and chemical arts are used herein to describe the compound of this invention. In general, the amino acid abbreviations follow the IUPAC-IUB Joint Commission on Biochemical Nomenclature as described in Eur. J. Biochem., 158, 9 (1984).
C1-6alkyl as applied herein means an optionally substituted alkyl group of 1 to 6 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, n-pentyl, isopentyl, neopentyl and hexyl and the simple aliphatic isomers thereof.
Any C1-6 alkyl may be optionally substituted with the group Rx, which may be on any carbon atom that results in a stable structure and is available by conventional synthetic techniques. Suitable groups for Rx are C1-4alkyl, ORxe2x80x3, SRxe2x80x3, C1-4alkylsulfonyl, C1-4alkylsulfoxyl, xe2x80x94CN, N(Rxe2x80x3)2, CH2N(Rxe2x80x3)2, xe2x80x94NO2, xe2x80x94CF3, xe2x80x94CO2Rxe2x80x3, xe2x80x94CON(Rxe2x80x3)2, xe2x80x94CORxe2x80x3, xe2x80x94NRxe2x80x3C(O)Rxe2x80x3, F, Cl, Br, I, or CF3S(O)rxe2x80x94, wherein r is 0, 1 or 2 and Rxe2x80x3 is H or C1-6alkyl.
Certain radical groups are abbreviated herein. t-Bu refers to the tertiary butyl radical, Boc refers to the t-butyloxycarbonyl radical, Fmoc refers to the fluorenylmethoxycarbonyl radical, Ph refers to the phenyl radical, Cbz refers to the benzyloxycarbonyl radical, Bn refers to the benzyl radical, Me refers to methyl, Et refers to ethyl, Ac refers to acetyl, Alk refers to C1-4alkyl, Nph refers to 1- or 2-naphthyl and cHex refers to cyclohexyl. Tet refers to 5-tetrazolyl.
Certain reagents are abbreviated herein. DCC refers to dicyclohexylcarbodiimide, DMAP refers to dimethylaminopyridine, DIEA refers to diisopropylethyl amine, EDC refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, hydrochloride. HOBt refers to 1-hydroxybenzotriazole, THP refers to tetrahydrofuiran, DIEA refers to diisopropylethylamine, DEAD refers to diethyl azodicarboxylate, PPh3 refers to triphenylphosphine, DIAD refers to diisopropyl azodicarboxylate, DME refers to dimethoxyethane, DMF refers to dimethylformamide, NBS refers to N-bromosuccinimide, Pd/C refers to a palladium on carbon catalyst, PPA refers to polyphosphoric acid, DPPA refers to diphenylphosphoryl azide, BOP refers to benzotriazol-1-yloxy-tris(dimethyl-amino)phosphonium hexafluorophosphate, HF refers to hydrofluoric acid, TEA refers to triethylamine, TFA refers to trifluoroacetic acid, PCC refers to pyridinium chlorochromate.
Compound of the formula (I) may be prepared by the methods described in Bondinell et al., PCT Publication No. WO 97/01540 (International Application No. PCT/US96/11108), published Jan. 16, 1997, the entire disclosure of which is incorporated herein by reference.
Additionally, the compound of formula (I) is prepared by methods analogous to those described in the schemes that are detailed hereinafter.
a) 10% Pd/C, HOAc; b) SOCl2, toluene; c) AlCl3, CH2Cl2
Scheme I details the preparation of an intermediate useful in the preparation of formula (I) compound.
a) LiN(TMS)2, ethyl bromoacetate; b) Jones reagent, OsO4; c) H2, 10% Pd/C, HOAC; d) C2O2Cl2, DMF; e) AlCl3, CH2Cl2, RT; f) H2, 10% Pd/C, HOAC
Scheme II also details the preparation of an intermediate useful in the preparation of formula (I) compound.
(a) EtOAc/LiHMDS, THF; (b) H2, 10% Pd/C, conc. HCl, AcOH; (c) EtSH, AlCl3, CH2Cl2; (d) 2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-ethanol, diisopropyl azodicarboxylate, (Ph)3P; (e) 1.0 N LiOH, EtOH; HCl.
Scheme III details the preparation of a formula (I) compound. Reaction of III-1 (which is a Scheme I-3 compound) in an aldol-type reaction with the enolate of ethyl acetate, which can be generated from ethyl acetate on exposure to an appropriate amide base, for instance lithium diisopropylamide (LDA) or lithium bis(trimethylsilyl)amide (LiHMDS), gives III-2. Frequently. THF is the solvent of choice for an aldol reaction, although THF in the presence of various additives, for instance HMPA or TMEDA, is often used. Reduction of III-2 to give III-3 (which is a Scheme II-6 compound) can be accomplished by hydrogenolysis over an appropriate catalyst, for example palladium metal on activated carbon. (Pd/C), in an appropriate solvent, such as acetic acid, in the presence of a mineral acid such as HCl. Alternatively, this reduction can be accomplished by treatment of III-2 with triethylsilane in the presence of boron trifluoride etherate by the general method of Orfanopoulos and Smonou (Synth. Commun. 1988, 833). Removal of the methyl ether of III-3 to give III-4 can be accomplished with BBr3 in an inert solvent, for example CH2Cl2, or by reaction with ethanethiol and AlCl3 in an inert solvent, preferably CH2Cl2. Other useful methods for removal of a methyl ether are described in Greene, xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d (published by John Wiley and Sons). Compound 4 of Scheme 3 (III-4) is reacted with 2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-ethanol to afford III-5. The reaction is mediated by the complex formed between diisopropyl azodicarboxylate and triphenylphosphine, and is conducted in an aprotic solvent, for instance THF, CH2Cl2, or DMF. The ethyl ester of III-5 is hydrolyzed using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the intermediate carboxylate salt is acidified with a suitable acid, for instance TFA or HCl, to afford the carboxylic acid III-6. Alternatively, the intermediate carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxylic acid can be prepared by methods well-known to those of skill in the art.
(a) PhOH, Cu, K2CO3; (b) sulfur, morpholine; (c) KOH, H2O, i-PrOH; (d) SOCl2, benzene; (e) AlCl3, CH2Cl2; (f) EtOAc, LiN(TMS)2, TMEDA, THF; (g) Et3SiH, BF3. OEt2, CH2Cl2; (h) H2, Pd/C, EtOH; (i) BBr3, CH2Cl2.
Commercially available 2-fluoro-4-methoxyacetophenone (IV-1) reacts with an alcohol, for example phenol, in the presence of copper metal and a suitable base, for instance K2CO3, to afford the diaryl ether IV-2. On treatment with sulfur and an appropriate primary or secondary amine, preferably morpholine, according to the general method of Harris (J. Med. Chem. 1982, 25, 855), IV-2 is converted to IV-3 in a classical Willgerodt-Kindler reaction. The thioamide thus obtained is hydrolyzed to the corresponding carboxylic acid IV-4 by reaction with an alkali metal hydroxide, suitably KOH, in an aqueous alcoholic solvent, such as aqueous MeOH, EtOH, or i-PrOH. Carboxylic acid IV-4 is converted to the corresponding acid chloride by reaction with either SOCl2 or oxalyl chloride according to conditions well-known to those of skill in the art. Treatment of this acid chloride with an appropriate Friedel-Crafts catalyst, such as AlCl3 or SnCl4, in an inert solvent, such as CH2Cl2 or CS2, provides the cyclic ketone IV-5. Alternatively, acid IV-4 can be converted directly to ketone IV-5 under acidic conditions, for example with polyphosphoric acid. Reaction of IV-5 in an aldolxe2x80x94type reaction with the enolate of ethyl acetate, which can be generated from ethyl acetate on exposure to an appropriate amide base, for instance lithium diisopropylamide (LDA) or lithium bis(trimethylsilyl)amide (LiHMDS), gives IV-6. Frequently, THF is the solvent of choice for an aldol reaction, although THF in the presence of various additives, for instance HMPA or TMEDA, is often used. Reduction of IV-6 to give IV-7 can be accomplished by treatment of IV-6 with triethylsilane in the presence of boron trifluoride etherate by the general method of Orphanopoulos and Smonu (Synth. Commun. 1988, 833). Any olefinic by-products that result from elimination of the alcohol are reduced by hydrogenation over an appropriate catalyst, for example palladium metal on activated carbon (Pd/C), in an appropriate solvent, such as MeOH or EtOH. Alternatively, the reduction of IV-6 to give IV-7 can be accomplished by hydrogenolysis in the presence of a mineral acid such as HCl. Typically, this reaction is catalyzed by Pd/C, and is optimally conducted in acetic acid. Removal of the methyl ether of IV-7 to give IV-8 can be accomplished with BBr3 in an inert solvent, for example CH2Cl2, or by reaction with ethanethiol and AlCl3 in an inert solvent, preferably CH2Cl2. Other useful methods for removal of a methyl ether are described in Greene, xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d (published by John Wiley and Sons). IV-8 is subsequently converted to formula (I) compound following the procedure outlined in Scheme III.
Acid addition salts of the compound are prepared in a standard manner in a suitable solvent from the parent compound and an excess of an acid, such as hydrochloric, hydrobromic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic or methanesulfonic. Certain of the compound form inner salts or zwitterions which may be acceptable. Cationic salts are prepared by treating the parent compound with an excess of an alkaline reagent, such as a hydroxide, carbonate or alkoxide, containing the appropriate cation; or with an appropriate organic amine. Cations such as Li+, Na+, K+, Ca++, Mg++ and NH4+ are specific examples of cations present in pharmaceutically acceptable salts.
This invention also provides a pharmaceutical composition which comprises a compound according to formula (I) and a pharmaceutically acceptable carrier. Accordingly, the compound of formula (I) may be used in the manufacture of a medicament. Pharmaceutical compositions of the compound of formula (I) prepared as hereinbefore described may be formulated as solutions or lyophilized powders for parenteral administration. Powders may be reconstituted by addition of a suitable diluent or other pharmaceutically acceptable carrier prior to use. The liquid formulation may be a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline solution, standard 5% dextrose in water or buffered sodium or ammonium acetate solution. Such formulation is especially suitable for parenteral administration, but may also be used for oral administration or contained in a metered dose inhaler or nebulizer for insufflation. It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxy cellulose, acacia, polyethylene glycol, mannitol, sodium chloride or sodium citrate.
Alternately, these compound may be encapsulated, tableted or prepared in a emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition, or to facilitate preparation of the composition. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline and water. The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage unit. The pharmaceutical preparations are made following the conventional techniques of pharmacy involving milling, mixing, granulating, and compressing, when necessary, for tablet forms; or milling, mixing and filling for hard gelatin capsule forms. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid formulation may be administered directly p.o. or filled into a soft gelatin capsule.
For rectal administration, the compound of this invention may also be combined with excipients such as cocoa butter, glycerin, gelatin or polyethylene glycols and molded into a suppository.
The compound described herein are antagonists of the vitronectin receptor, and are useful for treating diseases wherein the underlying pathology is attributable to ligand or cell which interacts with the vitronectin receptor. For instance, these compound are useful for the treatment of diseases wherein loss of the bone matrix creates pathology. Thus, the instant compound are useful for the treatment of ostoeporosis, hyperparathyroidism, Paget""s disease, hypercalcemia of malignancy, osteolytic lesions produced by bone metastasis, bone loss due to immobilization or sex hormone deficiency. The compound of this invention are also believed to have utility as antitumor, anti-angiogenic, antiinflammatory and anti-metastatic agents, and be useful in the treatment of atherosclerosis and restenosis.
The compound is administered either orally or parenterally to the patient, in a manner such that the concentration of drug is sufficient to inhibit bone resorption, or other such indication. The pharmaceutical composition containing the compound is administered at an oral dose of between about 0.1 to about 50 mg/kg in a manner consistent with the condition of the patient. Preferably the oral dose would be about 0.5 to about 20 mg/kg. For acute therapy, parenteral administration is preferred. An intravenous infusion of the peptide in 5% dextrose in water or normal saline, or a similar formulation with suitable excipients, is most effective, although an intramuscular bolus injection is also useful. Typically, the parenteral dose will be about 0.01 to about 100 mg/kg; preferably between 0.1 and 20 mg/kg. The compound are administered one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg/kg/day. The precise level and method by which the compound are administered is readily determined by one routinely skilled in the art by comparing the blood level of the agent to the concentration required to have a therapeutic effect.
This invention further provides a method for treating osteoporosis or inhibiting bone loss which comprises administering stepwise or in physical combination a compound of formula (I) and other inhibitors of bone resorption, such as bisphosphonates (i.e., allendronate), hormone replacement therapy, anti-estrogens, or calcitonin. In addition, this invention provides a method of treatment using a compound of this invention and an anabolic agent, such as the bone morphogenic protein, iproflavone, useful in the prevention of bone loss and/or to increase bone mass.
Additionally, this invention provides a method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I) and an antineoplastic agent. Compound of the camptothecin analog class, such as topotecan, irinotecan and 9-aminocamptothecin, and platinum coordination complexes, such as cisplatin, ormaplatin and tetraplatin, are well known groups of antineoplastic agents. Compound of the camptothecin analog class are described in U.S. Pat. Nos. 5,004,758, 4,604,463, 4,473,692, 4,545,880, 4,342,776, 4,513,138, 4,399,276, EP Patent Application Publication Nos. 0 418 099 and 0 088 642, Wani, et al., J. Med. Chem., 1986, 29, 2358, Wani, et al., J. Med. Chem., 1980, 23, 554, Wani, et al., J. Med. Chem, 1987, 30, 1774, and Nitta, et al., Proc. 14th International Congr. Chemotherapy., 1985, Anticancer Section 1, 28, the entire disclosure of each which is hereby incorporated by reference. The platinum coordination complex, cisplatin, is available under the name Platinol(copyright) from Bristol Myers-Squibb Corporation. Useful formulations for cisplatin are described in U.S. Pat. Nos. 5,562,925 and 4,310,515, the entire disclosure of each which is hereby incorporated by reference.
In the method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I) and an antineoplastic agent, the platinum coordination compound, for example cisplatin, can be administered using slow intravenous infusion. The preferred carrier is a dextrose/saline solution containing mannitol. The dose schedule of the platinum coordination compound may be on the basis of from about 1 to about 500 mg per square meter (mg/m2) of body surface area per course of treatment. Infusions of the platinum coordiation compound may be given one to two times weekly, and the weekly treatments may be repeated several times. Using a compound of the camptothecin analog class in a parenteral administration, the course of therapy generally employed is from about 0.1 to about 300.0 mg/m2 of body surface area per day for about five consecutive days. Most preferably, the course of therapy employed for topotecan is from about 1.0 to about 2.0 mg/m2 of body surface area per day for about five consecutive days. Preferably, the course of therapy is repeated at least once at about a seven day to about a twenty-eight day interval.
The pharmaceutical composition may be formulated with both the compound of formula (I) and the antineoplastic agent in the same container, but formualtion in different containers is preferred. When both agents are provided in solution form, they can be contained in an infusion/injection system for simultaneous administration or in a tandem arrangement.
For convenient administration of the compound of formula (I) and the antineoplastic agent at the same or different times, a kit is prepared, comprising, in a single container, such as a box, carton or other container, individual bottles, bags, vials or other containers each having an effective amount of the compound of formula (I) for parenteral administration, as described above, and an effective amount of the antineoplastic agent for parenteral administration, as described above. Such kit can comprise, for example, both pharmaceutical agents in separate containers or the same container, optionally as lyophilized plugs, and containers of solutions for reconstitution. A variation of this is to include the solution for reconstitution and the lyophilized plug in two chambers of a single container, which can be caused to admix prior to use. With such an arrangement, the antineoplastic agent and the compound of this invention may be packaged separately, as in two containers, or lyophilized together as a powder and provided in a single container.
When both agents are provided in solution form, they can be contained in an infusion/injection system for simultaneous administration or in a tandem arrangement. For example, the compound of formula (I) may be in an i.v. injectable form, or infusion bag linked in series, via tubing, to the antineoplastic agent in a second infusion bag. Using such a system, a patient can receive an initial bolus-type injection or infusion of the compound of formula (I) followed by an infusion of the antineoplastic agent.
The compound may be tested in one of several biological assays to determine the concentration of compound which is required to have a given pharmacological effect.
Inhibition of Vitronectin Binding
Solid-Phase [3H]-SKandF-107260 Binding to xcex1vxcex23: Human placenta or human platelet xcex1vxcex23 (0.1-0.3 mg/mL) in buffer T (containing 2 mM CaCl2 and 1% octylglucoside) was diluted with buffer T containing 1 mM CaCl2, 1 mM MnCl2, 1 mM MgCl2 (buffer A) and 0.05% NaN3, and then immediately added to 96-well ELISA plates (Corning, New York, N.Y.) at 0.1 mL per well. 0.1-0.2 xcexcg of xcex1vxcex23 was added per well. The plates were incubated overnight at 4xc2x0 C. At the time of the experiment, the wells were washed once with buffer A and were incubated with 0.1 mL of 3.5% bovine serum albumin in the same buffer for 1 hr at room temperature. Following incubation the wells were aspirated completely and washed twice with 0.2 mL buffer A.
Compound were dissolved in 100% DMSO to give a 2 mM stock solution, which was diluted with binding buffer (15 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM CaCl2, 1 mM MnCl2, 1 mM MgCl2) to a final compound concentration of 100 xcexcM. This solution is then diluted to the required final compound concentration. Various concentrations of unlabeled antagonists (0.001-100 xcexcM) were added to the wells in triplicates, followed by the addition of 5.0 nM of [3H]-SKandF-107260 (65-86 Ci/mmol).
The plates were incubated for 1 hr at room temperature. Following incubation the wells were aspirated completely and washed once with 0.2 mL of ice cold buffer A in a well-to-well fashion. The receptors were solubilized with 0.1 mL of 1% SDS and the bound [3H]-SKandF-107260 was determined by liquid scintillation counting with the addition of 3 mL Ready Safe in a Beckman LS Liquid Scintillation Counter, with 40% efficiency. Nonspecific binding of [3H]-SKandF-107260 was determined in the presence of 2 xcexcM SKandF-107260 and was consistently less than 1% of total radioligand input. The IC50 (concentration of the antagonist to inhibit 50% binding of [3H]-SKandF-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from the LUNDON-2 program. The Ki (dissociation constant of the antagonist) was calculated according to the equation: Ki=IC50/(1+L/Kd), where L and Kd were the concentration and the dissociation constant of [3H]-SKandF-107260, respectively.
The compound of the present invention inhibits vitronectin binding to SKandF 107260 at a Ki of about 1.7 nanomolar.
Compound of this invention are also tested for in vitro and in vivo bone resorption in assays standard in the art for evaluating inhibition of bone formation, such as the pit formation assay disclosed in EP 528 587, which may also be performed using human osteoclasts in place of rat osteoclasts, and the ovarectomized rat model, described by Wronski et al., Cells and Materials 1991, Sup. 1, 69-74.
Vascular Smooth Muscle Cell Migration Assay
Rat or human aortic smooth muscle cells were used. The cell migration was monitored in a Transwell cell culture chamber by using a polycarbonate membrane with pores of 8 um (Costar). The lower surface of the filter was coated with vitronectin. Cells were suspended in DMEM supplemented with 0.2% bovine serum albumin at a concentration of 2.5-5.0xc3x97106 cells/mL, and were pretreated with test compound at various concentrations for 20 min at 20xc2x0 C. The solvent alone was used as control. 0.2 mL of the cell suspension was placed in the upper compartment of the chamber. The lower compartment contained 0.6 mL of DMEM supplemented with 0.2% bovine serum albumin. Incubation was carried out at 37xc2x0 C. in an atmosphere of 95% air/5% CO2 for 24 hr. After incubation, the non-migrated cells on the upper surface of the filter were removed by gentle scraping. The filter was then fixed in methanol and stained with 10% Giemsa stain. Migration was measured either by a) counting the number of cells that had migrated to the lower surface of the filter or by b) extracting the stained cells with 10% acetic acid followed by determining the absorbance at 600 nM.
Thyroparathyroidectomized Rat Model
Each experimental group consists of 5-6 adult male Sprague-Dawley rats (250-400 g body weight). The rats are thyroparathyroidectomized (by the vendor, Taconic Farms) 7 days prior to use. All rats receive a replacement dose of thyroxine every 3 days. On receipt of the rats, circulating ionized calcium levels are measured in whole blood immediately after it has been withdrawn by tail venipuncture into heparinized tubes. Rats are included if the ionized Ca level (measured with a Ciba-Corning model 634 calcium pH analyzer) is  less than 1.2 mM/L. Each rat is fitted with an indwelling venous and arterial catheter for the delivery of test material and for blood sampling respectively. The rats are then put on a diet of calcium-free chow and deionized water. Baseline Ca levels are measured and each rat is administered either control vehicle or human parathyroid hormone 1-34 peptide (hPTH1-34, dose 1.25 ug/kg/h in saline/0.1% bovine serum albumin, Bachem, Ca) or a mixture of hPTH1-34 and test material, by continuous intravenous infusion via the venous catheter using an external syringe pump. The calcemic response of each rat is measured at two-hourly intervals during the infusion period of 6-8 hours.
Human Osteoclast Resorption and Adhesion Assays
Pit resorption and adhesion assays have been developed and standardized using normal human osteoclasts derived from osteoclastoma tissue. Assay 1 was developed for the measurement of osteoclast pit volumes by laser confocal microscopy. Assay 2 was developed as a higher throughput screen in which collagen fragments (released during resorption) are measured by competitve ELISA.
Assay 1 (Using Laser Confocal Microscopy)
Aliquots of human osteoclastoma-derived cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37xc2x0 C. and washed xc3x971 in RPMI-1640 medium by centrifugation (1000 rpm, 5 mins at 4xc2x0 C.).
The medium is aspirated and replaced with murine anti-HLA-DR antibody then diluted 1:3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice and mixed frequently.
The cells are washed xc3x972 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mins at 4xc2x0 C.) and the cells are then transferred to a sterile 15 ml centrifuge tube. The number of mononuclear cells are enumerated in an improved Neubauer counting chamber.
Sufficient magnetic beads (5/mononuclear cell), coated with goat anti-mouse IgG (Dynal, Great Neck, N.Y.) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removed by immobilizing the beads on a magnet and is replaced with fresh medium.
The beads are mixed with the cells and the suspension is incubated for 30 mins on ice. The suspension is mixed frequently.
The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube.
Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts. This wash process is repeatedxc3x9710. The bead-coated cells are discarded.
The viable osteoclasts are enumerated in a counting chamber, using fluorescein diacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber.
The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1.7 g/liter of sodium bicarbonate.
3 ml aliquots of the cell suspension (per compound treatment) are decanted into 15 ml centrifuge tubes. The cells are pelleted by centrifugation.
To each tube, 3 ml of the appropriate compound treatment are added (diluted to 50 uM in the EMEM medium). Also included are appropriate vehicle controls, a positive control (anti-vitronectin receptor murine monoclonal antibody [87MEM1] diluted to 100 ug/ml) and an isotype control (IgG2a diluted to 100 ug/ml). The samples are incubated at 37xc2x0 C. for 30 mins.
0.5 ml aliquots of the cells are seeded onto sterile dentine slices in a 48-well plate and incubated at 37xc2x0 C. for 2 hours. Each treatment is screened in quadruplicate.
The slices are washed in six changes of warm PBS (10 ml/well in a 6-well plate) and then placed into fresh medium containing the compound treatment or control samples. The samples are incubated at 37xc2x0 C. for 48 hours.
Tartrate Resistant Acid Phosphatase (TRAP) Procedure (Selective Stain for Cells of the Osteoclast Lineage)
The bone slices containing the attached osteoclasts are washed in phosphate buffered saline and fixed in 2% gluteraldehyde (in 0.2M sodium cacodylate) for 5 mins.
They are then washed in water and are incubated for 4 minutes in TRAP buffer at 37xc2x0 C. (0.5 mg/ml naphthol AS-BI phosphate dissolved in N,N-dimethylfornamide and mixed with 0.25 M citrate buffer (pH 4.5), containing 10 mM sodium tartrate.
Following a wash in cold water the slices are immersed in cold acetate buffer (0.1 M, pH 6.2) containing 1 mg/ml fast red garnet and incubated at 4xc2x0 C. for 4 minutes.
Excess buffer is aspirated, and the slices are air dried following a wash in water.
The TRAP positive osteoclasts (brick red/ purple precipitate) are enumerated by bright-field microscopy and are then removed from the surface of the dentine by sonication.
Pit volumes are determined using the Nikon/Lasertec ILM21W confocal microscope.
Assay 2 (Using an ELISA Readout)
The human osteoclasts are enriched and prepared for compound screening as described in the initial 9 steps of Assay 1. For clarity, these steps are repeated hereinbelow.
Aliquots of human osteoclastoma-derived cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37xc2x0 C. and washed xc3x971 in RPMI-1640 medium by centrifugation (1000 rpm, 5 mins at 4xc2x0 C.).
The medium is aspirated and replaced with murine anti-HLA-DR antibody then diluted 1:3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice and mixed frequently.
The cells are washed xc3x972 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mins at 4xc2x0 C.) and the cells are then transferred to a sterile 15 ml centrifuge tube. The number of mononuclear cells are enumerated in an improved Neubauer counting chamber.
Sufficient magnetic beads (5/mononuclear cell), coated with goat anti-mouse IgG (Dynal, Great Neck, N.Y.) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removed by immobilizing the beads on a magnet and is replaced with fresh medium.
The beads are mixed with the cells and the suspension is incubated for 30 mins on ice. The suspension is mixed frequently.
The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube.
Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts. This wash process is repeated xc3x9710. The bead-coated cells are discarded.
The viable osteoclasts are enumerated in a counting chamber, using fluorescein diacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber.
The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1.7 g/liter of sodium bicarbonate.
In contrast to the method desribed above in Assay 1, the compound are screened at 4 doses to obtain an IC50, as outlined below:
The osteoclast preparations are preincubated for 30 minutes at 37xc2x0 C. with test compound (4 doses) or controls.
They are then seeded onto bovine cortical bone slices in wells of a 48-well tissue culture plate and are incubated for a further 2 hours at 37xc2x0 C.
The bone slices are washed in six changes of warm phosphate buffered saline (PBS), to remove non-adherent cells, and are then returned to wells of a 48 well plate containing fresh compound or controls.
The tissue culture plate is then incubated for 48 hours at 37xc2x0 C.
The supernatants from each well are aspirated into individual tubes and are screened in a competitive ELISA that detects the c-telopeptide of type I collagen which is released during the resorption process. This is a commercially available ELISA (Osteometer, Denmark) that contains a rabbit antibody that specifically reacts with an 8-amino acid sequence (Glu-Lys-Ala-His- Asp-Gly-Gly-Arg) that is present in the carboxy-terminal telopeptide of the a1-chain of type I collagen. The results are expressed as % inhibition of resorption compared to a vehicle control.
Human Osteoclast Adhesion Assay
The human osteoclasts are enriched and prepared for compound screening as described above in the inital 9 steps of Assay 1. For clarity, these steps are repeated hereinbelow.
Aliquots of human osteoclastoma-derived cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37xc2x0 C. and washed xc3x971 in RPMI-1640 medium by centrifugation (1000 rpm, 5 mins at 4xc2x0 C.).
The medium is aspirated and replaced with murine anti-HLA-DR antibody then diluted 1:3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice and mixed frequently.
The cells are washed xc3x972 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mins at 4xc2x0 C.) and the cells are then transferred to a sterile 15 ml centrifuge tube. The number of mononuclear cells are enumerated in an improved Neubauer counting chamber.
Sufficient magnetic beads (5/mononuclear cell), coated with goat anti-mouse IgG (Dynal, Great Neck, N.Y.) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removed by immobilizing the beads on a magnet and is replaced with fresh medium.
The beads are mixed with the cells and the suspension is incubated for 30 mins on ice. The suspension is mixed frequently.
The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube.
Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts, This wash process is repeatedxc3x9710. The bead-coated cells are discarded.
The viable osteoclasts are enumerated in a counting chamber, using fluorescein diacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber.
The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1.7 g/liter of sodium bicarbonate.
Osteoclastoma-derived osteoclasts are preincubated with compound (4 doses) or controls at 37xc2x0 C. for 30 minutes.
The cells are then seeded onto osteopontin-coated slides (human or rat osteopontin, 2.5 ug/ml) and incubated for 2 hours at 37xc2x0 C.
Non adherent cells are removed by washing the slides vigorously in phosphate buffered saline and the cells remaining on the slides are fixed in acetone.
The osteoclasts are stained for tattrate-resistant acid phosphatase (TRAP), a selective marker for cells of this phenotype (see steps 15-17), and are enumerated by light microscopy. The results are expressed as % inhibition of adhesion compared to a vehicle control.
Cell Adhesion Assay
Cells and Cell Culture
Human embryonic kidney cells (HEK293 cells) were obtained from ATCC (Catalog No. CRL 1573). Cells were grown in Earl""s minimal essential medium (EMEM) medium containing Earl""s salts, 10% fetal bovine serum, 1% glutamine and 1% Penicillin-Steptomycin.
Constructs and Transfections
A 3.2 kb EcoRI-KpnI fragment of the xcex1v subunit and a 2.4 kb XbaI-XhoI fragment of the xcex23 subunit were inserted into the EcoRI-EcoRV cloning sites of the pCDN vector (Aiyar et al., 1994 ) which contains a CMV promoter and a G418 selectable marker by blunt end ligation. For stable expression, 80xc3x97106 HEK 293 cells were electrotransformed with xcex1v+xcex23 constructs (20 xcexcg DNA of each subunit) using a Gene Pulser (Hensley et al., 1994 ) and plated in 100 mm plates (5xc3x97105 cells/plate). After 48 hr, the growth medium was supplemented with 450 xcexcg/mL Geneticin (G418 Sulfate, GIBCO-BRL, Bethesda, Md.). The cells were maintained in selection medium until the colonies were large enough to be assayed.
Immunocytochemical Analysis of Transfected Cells
To determine whether the HEK 293 transfectants expressed the vitronectin receptor, the cells were immobilized on glass microscope slides by centrifugation, fixed in acetone for 2 min at room temperature and air dried. Specific reactivity with 23C6, a monoclonal antibody specific for the xcex1vxcex23 complex was demonstrated using a standard indirect immunofluorescence method.
Cell Adhesion Studies
Corning 96-well ELISA plates were precoated overnight at 4xc2x0 C. with 0.1 mL of human vitronectin (0.2 xcexcg/mL in RPMI medium). At the time of the experiment, the plates were washed once with RPMI medium and blocked with 3.5% BSA in RPMI medium for 1 hr at room temperature. Transfected 293 cells were resuspended in RPMI medium, supplemented with 20 mM Hepes, pH 7.4 and 0.1% BSA at a density of 0.5xc3x97106 cells/mL. 0.1 mL of cell suspension was added to each well and incubated for 1 hr at 37xc2x0 C., in the presence or absence of various xcex1vxcex23 antagonists. Following incubation, 0.025 mL of a 10% formaldehyde solution, pH 7.4, was added and the cells were fixed at room temperature for 10 min. The plates were washed 3 times with 0.2 mL of RPMI medium and the adherent cells were stained with 0.1 mL of 0.5% toluidine blue for 20 min at room temperature. Excess stain was removed by extensive washing with deionized water. The toluidine blue incorporated into cells was eluted by the addition of 0.1 mL of 50% ethanol containing 50 mM HCl. Cell adhesion was quantitated at an optical density of 600 nm on a microtiter plate reader (Titertek Multiskan MC, Sterling, Va.).
Solid-Phase xcex1vxcex25 Binding Assay
The vitronectin receptor xcex1vxcex25 was purified from human placenta. Receptor preparation was diluted with 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM CaCl2, 1 mM MnCl2, 1 mM MgCl2 (buffer A) and was immediately added to 96-well ELISA plates at 0.1 ml per well. 0.1-0.2 xcexcg of xcex1vxcex23 was added per well. The plates were incubated overnight at 4xc2x0 C. At the time of the experiment, the wells were washed once with buffer A and were incubated with 0.1 ml of 3.5% bovine serum albumin in the same buffer for 1 hr at room temperature. Following incubation the wells were aspirated completely and washed twice with 0.2 ml buffer A.
In a [3H]-SKandF-107260 competition assay, various concentrations of unlabeled antagonists (0.001-100 xcexcM) were added to the wells, followed by the addition of 5.0 nM of [3H]-SKandF-107260. The plates were incubated for 1 hr at room temperature. Following incubation the wells were aspirated completely and washed once with 0.2 ml of ice cold buffer A in a well-to-well fashion. The receptors were solubilized with 0.1 ml of 1% SDS and the bound [3H]-SKandF-107260 was determined by liquid scintillation counting with the addition of 3 ml Ready Safe in a Beckman LS 6800 Liquid Scintillation Counter, with 40% efficiency. Nonspecific binding of [3H]-SKandF-107260 was determined in the presence of 2 xcexcM SKandF-107260 and was consistently less than 1% of total radioligand input. The IC50 (concentration of the antagonist to inhibit 50% binding of [3H]-SKandF-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from the LUNDON-2 program. The Ki (dissociation constant of the antagonist) was calculated according to Cheng and Prusoff equation: Ki=IC50/(1+L/Kd), where L and Kd were the concentration and the dissociation constant of [3H]-SKandF-107260, respectively.
Purification of GPIIb-IIIa
Ten units of outdated, washed human platelets (obtained from Red Cross) were lyzed by gentle stirring in 3% octylglucoside, 20 mM Tris-HCl, pH 7.4, 140 mM NaCl, 2 mM CaCl2 at 40xc2x0 C. for 2 h. The lysate was centrifuged at 100,000 g for 1 h. The supernatant obtained was applied to a 5 mL lentil lectin sepharose 4B column (E.Y. Labs) preequilibrated with 20 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaCl2, 1% octylglucoside (buffer A). After 2 h incubation, the column was washed with 50 mL cold buffer A. The lectin-retained GPIIb-IIIa was eluted with buffer A containing 10% dextrose. All procedures were performed at 4xc2x0 C. The GPIIb-IIIa obtained was  greater than 95% pure as shown by SDS polyacrylamide gel electrophoresis.
Incorporation of GPIIb-IIIa in Liposomes
A mixture of phosphatidylserine (70%) and phosphatidylcholine (30%) (Avanti Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen. Purified GPIIb-IIIa was diluted to a final concentration of 0.5 mg/mL and mixed with the phospholipids in a protein:phospholipid ratio of 1:3 (w:w). The mixture was resuspended and sonicated in a bath sonicator for 5 min. The mixture was then dialyzed overnight using 12,000-14,000 molecular weight cutoff dialysis tubing against a 1000-fold excess of 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaCl2 (with 2 changes). The GPIIb-IIIa-containing liposomes wee centrifuged at 12,000 g for 15 min and resuspended in the dialysis buffer at a final protein concentration of approximately 1 mg/mL. The liposomes were stored at xe2x88x9270xc2x0 C. until needed.
Competitive Binding to GPIIb-IIIa
The binding to the fibrinogen receptor (GPIIB-IIIa) was assayed by an indirect competitive binding method using [3H]-SKandF-107260 as an RGD-type ligand. The binding assay was performed in a 96-well filtration plate assembly (Millipore Corporation, Bedford, Mass.) using 0.22 um hydrophilic durapore membranes. The wells were precoated with 0.2 mL of 10 xcexcg/mL polylysine (Sigma Chemical Co., St. Louis, Mo.) at room temperature for 1 h to block nonspecific binding. Various concentrations of unlabeled benzazepines were added to the wells in quadruplicate. [3H]-SKandF-107260 was applied to each well at a final concentration of 4.5 nM, followed by the addition of 1 xcexcg of the purified platelet GPIIb-IIIa-containing liposomes. The mixtures were incubated for 1 h at room temperature. The GPIIb-IIIa-bound [3H]-SKandF-107260 was seperated from the unbound by filtration using a Millipore filtration manifold, followed by washing with ice-cold buffer (2 times, each 0.2 mL). Bound radioactivity remaining on the filters was counted in 1.5 mL Ready Solve (Beckman Instruments, Fullerton, Calif.) in a Beckman Liquid Scintillation Counter (Model LS6800), with 40% efficiency. Nonspecific binding was determined in the presence of 2 xcexcM unlabeled SKandF-107260 and was consistently less than 0.14% of the total radioactivity added to the samples. All data points are the mean of quadruplicate determinations.
Competition binding data were analyzed by a nonlinear least-squares curve fitting procedure. This method provides the IC50 of the antagonists (concentration of the antagonist which inhibits specific binding of [3H]-SKandF-107260 by 50% at equilibrium). The IC50 is related to the equilibrium dissociation constant (Ki) of the antagonist based on the Cheng and Prusoff equation: Ki=IC50/(1+L/Kd), where L is the concentration of [3H]-SKandF-107260 used in the competitive binding assay (4.5 nM), and Kd is the dissociation constant of [3H]-SKandF-107260 which is 4.5 nM as determined by Scatchard analysis.
The efficacy of the compound of formula (I) alone or in combination with an antineoplastic agent may be determined using several transplantable mouse tumor models. See U.S. Pat. Nos. 5,004,758 and 5,633,016 for details of these models
The examples which follow are intended in no way to limit the scope of this invention, but are provided to illustrate how to make and use the compound of this invention. Many other embodiments will be readily apparent to those skilled in the art.


An emergency lighting system with central batteries power remote LED fixtures and exit signs, with all components integrated throughout a single or multiple buildings. Status reports are available on demand and the system will generate written records required by the Life Safety Code. A wide range of fixtures optimized for emergency lighting are available for interior and exterior applications and all can be monitored via internet, with report displays available on any digital device. Easy access to batteries, automated testing, high lumen outputs, efficient optics and long lamp life ensure the lowest possible life cycle costs.

1. Technical Field
The disclosure relates to lighting systems with backup power supplies and, more particularly, to emergency lighting systems with centralized backup power, periodic testing systems and methods, and reporting and monitoring systems and methods.
2. Background Information
Emergency lighting products in the USA and other countries are required to be installed in all commercial, industrial and other buildings with public access. Regulations controlling operation and performance of emergency lighting products in the USA are published by the National Fire Protection Association Life Safety Code No. 101, Chapter 7, Means of Egress (LSC 101). Periodic testing of such equipment is required and described in the LSC 101, and is enforced by the Authority Having Jurisdiction (AHJ). The Life Safety Code names four key elements that must be included in every emergency lighting product using batteries to provide stored energy for emergency lighting following loss of main building power:
1. Battery capacity
2. Lamp operation
3. Transfer switch operation
4. Battery charger operation
Testing of all four items is to be performed monthly by simulating a brief loss of main building power, and annually starting 12 months after installation, when emergency operation under battery power for the full rated time is required, commonly 90 minutes but may be longer for certain building occupancies. Results of these tests are to be recorded in permanent written record, to be available for inspection by the AHJ.
Three approved methods for performing the four tests are described in LSC 101, including one describing a computer-based, automated system to minimize the amount of manual labor required, to maintain a history of tests completed, and to produce written test results on demand. Paragraph 126.96.36.199 of LSC 101 contains the following wording: Computer based, self-testing/self diagnostic battery-operated emergency lighting equipment capable of providing a report of the history of tests and failures at all times.
Given the number of fixtures required in typical commercial and industrial buildings, a manual inspection process meeting the requirements for the four tests identified above is labor intensive. With conventional emergency lighting fixtures, monthly and annual tests are accomplished by activation of emergency lighting by pressing a Push to Test (PIT) momentary switch on every fixture. This simulates loss of normal power for as long as the PIT is held down. LSC 101 requires this operation for 30 seconds on every fixture, every month. Results from the test are to be recorded in writing, and the report must be available for inspection by the AHJ at any time. Completion of these tests and reports is an onerous procedure and is rarely accomplished correctly or completely by facility or maintenance managers.
Emergency lighting is required to operate upon loss of normal building power typically supplied by a utility as Alternating Current (AC). Power for emergency lighting must be provided from an independent source, usually batteries, or an electrical generator. The two most common forms of emergency lighting in use today are:
1. Unit equipment, defined as an emergency lighting fixture with a self-contained battery pack, a battery charger and a transfer switch to supply Direct Current (DC) electrical power from the battery for emergency lighting operation upon loss of building power.
2. A central or smaller inverter, defined as a power system that typically includes multiple batteries, a battery charger, a control circuit to convert power supplied by batteries from DC into AC, and a transfer switch to supply this AC power for emergency operation of any standard lighting fixture connected to the inverter.
Both forms rely on stored energy from batteries. When normal power is supplied the batteries are connected to a charger which supplies constant current to ensure they are maintained at full capacity. Upon loss of building power, the transfer switch connects batteries to lamps in the emergency fixtures. In that event, the Life Safety Code stipulates that the amount of battery energy available must be sufficient to operate all emergency lighting lamps for a minimum of 90 minutes, after which the battery terminal voltage must be no less than 87.5% of the rated battery voltage, with the lamps remaining on. This performance requirement is strictly regulated under Underwriters Laboratories (UL) Standard 924 for Emergency Lighting and Power Equipment, which is based on criteria enumerated in LSC 101.
Operating conditions and maintenance problems concerning the two forms are as follows:
Unit Equipment: Batteries most commonly in use today for unit equipment emergency lighting products are Sealed Lead Acid (SLA), Nickel Cadmium (NICAD) or Nickel Metal Hydride (NIMH). Over time, the amount of stored energy in all types of battery will decrease, to the point when a fully charged battery is no longer able to meet the minimum power requirements for emergency operation, and they must then be immediately replaced. The typical average life of SLA batteries is 4 years; that of NICAD or NIMH batteries may be slightly longer. If the ambient temperature around a battery is elevated, as frequently happens in certain building installations, battery life may be less than 4 years.
LSC 101 specifies that all emergency lighting equipment be tested monthly for correct operation of the battery and the three other functions listed above, following a brief battery discharge. Then, every year starting 12 months after installation the fixtures must be tested with a battery discharge of 90 minutes (or more if specified for certain occupancies) at full load. If any of the three functions listed fails, or any battery fails to meet the 87.5% voltage minimum after 90 minutes, the fixture is in violation of the Life Safety Code, must be repaired or replaced to correct the fault, and retested to confirm proper operation. Results from these tests and/or corrections are to be recorded in writing, and the report must be available for inspection by the AHJ at any time. It is therefore important for the facility owner to be vigilant in reviewing the status of emergency lamps and the ability of batteries to maintain the 90 minute discharge.
The problem of review and inspection of unit equipment has been recognized in the industry and products have been developed to perform automatic, self-testing diagnostics of batteries and lamps, typically using a color-coded warning light on the fixture, or in some cases by incorporating a transmitter using WIFI to transfer diagnostic data from multiple fixtures to a central site. While these solutions may help the facility manager to identify equipment with bad lamps or batteries, they are expensive to install and do nothing to reduce the cost of replacing components found to be faulty, which is by far the largest portion of total maintenance expense.
Replacement of batteries and lamps in unit equipment is labor intensive because each fixture with a failed component must be accessed (most often requiring a ladder or a portable lifting system for fixtures in high locations), dismantled, have wiring disconnected, the battery, lamps or other components of the charging mechanism or the transfer switch replaced with new components, followed by reconnection, reassembly and testing. In many products, the battery is part of a package that includes the charger and transfer switch (known as an "emergency ballast'/) which increases the value of the material requiring replacement. In addition, typical batteries or emergency ballasts are manufactured specifically for the fixture and are thus not readily available as a stock item from electrical distributors; they must be carefully identified by part number and purchased direct from the factory or through a specialized supply service.
As described, total cost for routine maintenance of unit equipment for any facility can be substantial, especially in cases where large quantities of fixtures are installed. For example, unit equipment in a typical high school may contain more than one thousand batteries.
Inverters: An inverter may be supplied in a wide range of power capacities, from small models that supply one or a small number of lighting fixture with emergency power, to central power systems that can supply all fixtures throughout a large building. All inverters include a battery charger, a transfer switch and for emergency operation, an electrical circuit to convert DC power supplied from batteries into AC power for the lighting fixtures. The converter circuits are relatively complex and especially in larger machines, expensive. Frequently, multiple batteries are installed in larger machines and connected in series to produce high DC voltages, from 48V to 120V or more. Because of the inherent danger associated with high DC voltages and complex wiring schemes, manufacturers of such equipment usually require installation and setup by factory trained mechanics. Maintenance requiring replacement of batteries is also required to be carried out by factory technicians or electrical contractors with specific experience or training with this type of equipment.
Typical fixtures connected to inverters are designed for general lighting under normal conditions and as such, the lumen output, lamp power and beam spread patters are not optimized for emergency lighting. LSC 101 stipulates that emergency lighting must produce uniform illumination at a specified brightness level from 5 to 50 times lower than general lighting, along a path of egress, therefore an elongated beam spread concentrated along the egress pathway is desirable. However general lighting fixtures are designed to produce lighting over as large an area as possible, and most produce generally circular patterns, not elongated beams. The high power lamps used in these fixtures require far more battery power than lamps designed for emergency lighting. General lighting fixtures with multiple lamps can be configured to operate with a reduced number of lamps for emergency operation, thus saving battery energy, but that cannot be done with single-lamp fixtures.
As a result, general lighting fixtures operate at higher power than is required by the Code, requiring larger, more expensive batteries compared to a central power system for fixtures designed specifically for emergency lighting. Also, more fixtures with circular lighting patterns are required, compared to those with optical means designed to produce an elongated beam spread.
Lamps in use for general lighting may operate an average of 4,380 hours per year (12 hrs/day for 365 days). The most common type of lamps in use today are fluorescent, with an average life of about 15,000 hours or less. Therefore, many lamps will require replacement every 3-4 years and those connected to the emergency circuit will require thorough inspection and regular replacement to maintain code compliance.

Emergency lighting monitoring and reporting system


A multi-band antenna (1) used in an electronic device and formed of a metallic sheet by defining holes therein, including a first radiating portion (30), a second radiating portion (31), a third radiating portion (32), a ground portion (2), and a coaxial transmission line (4). The first radiating portion, the ground portion and the coaxial transmission line cooperatively form a loop antenna operated at a higher frequency band of about 5.15–5.875 GHz. The second radiating portion, the ground portion and the coaxial transmission line cooperatively form a first inverted-F antenna operated at another higher frequency band of about 5.725–5.875 GHz. The third radiating portion, the ground portion and the coaxial transmission line cooperatively form a second inverted-F antenna operated at a lower frequency band of about 2.4–2.5 GHz.

1. Field of the Invention
The present invention relates generally to an antenna, and more particularly to a multi-band antenna for use with an electronic device.
2. Description of the Prior Art
The prosperous development of wireless communication industry brings various products and techniques for multi-band communication such that many new products have the performance for wireless communication so as to meet the consumers' demand. Such products as WLAN cards with antennas on/in them for use with a laptop computer or a personal digital assistance (PDA) are gaining popularity in wireless communication market. These cards benefit from multi-band antennas operated under IEEE 802.11a/b/g standard. In most cases, embedded multi-band antennas are arranged in an electronic device directly, rather than via a WLAN card. Whatever, a multi-band antenna with small size and high performance is essential and critical to achieve the purpose for wireless communication.
A series of dual-band antennas embedded within electronic devices are disclosed in U.S. patent application Publication No. 2003/0222823, including slot-slot antenna, PIFA-PIFA antenna, and PIFA-slot antenna, and so on. Take a general architecture of a PIFA-PIFA antenna for example. The dual-band antenna 700 comprises a first radiating element comprising components 702 and 703, and a second radiating element comprising components 704 and 708. The first and the second radiating elements are connected to a ground element 701. An antenna feed is preferably implemented using a coaxial transmission line 706, wherein an inner conductor 705 of the coaxial transmission line is connected to the first radiating element, and an outer conductor 707 of the coaxial transmission line is connected to the ground element 701. The antenna 700 operates in a lower frequency band of about 2.4 GHz to about 2.5 GHz under IEEE 802.11b/g and a higher frequency band of about 5.15 GHz to about 5.35 GHz under IEEE 802.11a. However, the antenna cannot be used in another frequency band of 5.75–5.825 GHz which is also under IEEE 802.11a standard. Moreover, the lower and the higher frequency bands of the antenna are narrow, which restrains the application of the antenna. Additionally, the second radiating element of the antenna is fed though coupling, rather than by the coaxial transmission line directly, which reversely affects the antenna gain.
Hence, in this art, a multi-band antenna with wide bandwidth to overcome the above-mentioned disadvantages of the prior art will be described in detail in the following embodiments.


A speaker tube structure formed of a semi-cylindrical first side wall portion and a planar second side wall portion, with the second side wall portion including speaker members mounted therewithin, having a tuning port orthogonally directed into the second side wall portion extending into the speaker tube cavity in a spaced adjacency relative to the tube axis. The tuning port is arranged to receive a plurality of inserts to permit tuning of the speaker tube.

1. Field of the Invention
The field of invention relates to speaker enclosure structure, and more particularly pertains to a new and improved tunable speaker tube wherein the same is arranged for effecting efficient reflection of sound within the tube cavity.
2. Description of the Prior Art
Speaker tubes typically of polygonal type configurations loose response within the enclose, wherein the instant invention attempts to overcome deficiencies of the prior art by providing for a semi-cylindrical tube structure arranged to enhance reflection of sound within the tube enclosure.
Prior art speaker tube structure is indicated in U.S. Pat. No. 4,757,548 to Fenner indicating a dome-shaped enclosure in contrast to the cylindrical construction of the instant invention, wherein U.S. Pat. Nos. 3,449,519; 5,025,473; and 4,122,302 are further examples of conventional speaker enclosure structure.


A heat-resistant separator for an electrochemical element in which the thickness of the separator is reduced while maintaining the balance between the short circuit resistance, resistance, electrolyte impregnation performance, and electrolyte retention performance of the separator. A separator for an electrochemical element includes beaten cellulose fibers, wherein the value obtained by dividing the average value for the distance between the center point of a cellulose stem fiber constituting part of the separator and the center point of another cellulose stem fiber nearest to said cellulose stem fiber by the thickness of the separator is 0.80 to 1.35.

Examples of electrochemical elements include aluminum electrolytic capacitors, conductive polymer aluminum solid electrolytic capacitors, conductive polymer hybrid aluminum electrolytic capacitors, electric double layer capacitors, lithium ion capacitors, lithium ion secondary batteries, and lithium primary batteries. Moreover, these electrochemical elements are adopted in number of fields including motor vehicle-related devices, digital devices, renewable energy-related devices such as wind power generation and solar power generation, and communication devices such as smart meter, and the applications thereof are expected to continuously expand in the future.
For example, in motor vehicles, a lithium ion secondary battery or an electric double layer capacitor is used for energy regeneration and an aluminum electrolytic capacitor is used for power supply to various electronic control units. Furthermore, in electric vehicles and hybrid vehicles which have attracted attention in recent years, a lithium ion secondary battery is used as a power source and the like, an electric double layer capacitor is used for energy regeneration and the like, and an aluminum electrolytic capacitor is used for control of a battery and a hybrid system serving as a power source and in an electronic control unit of direct current conversion from an external alternating current power source, and the like. Such electrochemical elements used in motor vehicles are required to exhibit high reliability since defects directly affect human life. In other words, these electrochemical elements are required to withstand vibration of the vehicle body, not to cause short circuit defect even when being used for a long time in a cold region or a high-temperature region, and also to hardly cause deterioration in performance.
In the circuit board used in electronic devices, there is a great demand for thinning and downsizing of electrochemical elements mounted. Moreover, electrochemical elements used in the power source of portable devices are also required to be used for a long time by one time of charge.
For this reason, electrochemical elements such as aluminum electrolytic capacitors mounted on a board are required to have a low height and a small size for the purpose of supplying power to the chips and CPUs and smoothing the alternating current.
In addition, lithium ion secondary batteries, which are often used as a power source of portable devices, are required to have a high capacity so as to be used for a long time while having a low height and to hardly cause deterioration in performance even when being repeatedly charged and discharged.
In association with such expansion of applications and improvement in performance of the devices used, electrochemical elements are required to be improved in performance such as charge and discharge characteristics and output characteristics, to exhibit further reliability, to be downsized, and the like more than ever.
As a separator for an electrochemical element, a cellulose separator, a microporous polyolefin membrane, a separator in which a synthetic fiber nonwoven fabric is coated with an inorganic substance, and the like are used.
As the microporous polyolefin membrane separator, a separator made of polypropylene or polyethylene is used.
As the cellulose separator, natural cellulose fibers or regenerated cellulose fibers are used. As the regenerated cellulose fibers, solvent-spun cellulose fibers are used.
As the separator in which a synthetic fiber nonwoven fabric is coated with an inorganic substance, those in which nonwoven fabrics obtained by forming synthetic fibers such as polyester fibers into sheets by a wet papermaking method are coated with inorganic powders are proposed.
The main role of a separator in an electrochemical element is the separation of a pair of electrodes and the retention of the electrolyte. In order to separate a pair of electrodes, the separator is required to exhibit high shielding properties. Moreover, in order to improve the shielding property, it is required to increase the density of the separator.
In addition, thinning of the separator is also required for the purpose of downsizing (decreasing the diameter and/or decreasing the height) the electrochemical element and increasing the capacity of the electrochemical element.
In order to thin the separator for an electrochemical element, the membrane thickness of microporous polyolefin membrane separator is required to be still more thinned. Those coated with inorganic powders such as aluminum oxide have been proposed for the purpose of preventing a decrease in heat resistance and short circuit resistance to be inevitably caused by thinning of membrane thickness.
Meanwhile, in order to thin cellulose separators, a method is adopted in which the thickness of separator at the time of manufacture (papermaking) is thinned or the separator after papermaking is thinned by a linear pressure applied using a press.
In order to improve the performance of separator, for example, the techniques of Patent Literatures 1 to 5 are disclosed.

Separator for electrochemical element and electrochemical element


A reflect antenna element having a receive antenna section and a transmit antenna section. Each section has a cavity, a conductive element in registration with the cavity, and a ground plane conductor having a slot. A strip conductor has portions thereof disposed over the slots and the ground planes conductor. The strip conductor and underlying ground plane conductor form a microstrip transmission line for coupling energy received by the receive antenna section to the transmit antenna section. The transmit antenna section and receive antenna section are configured to operate with orthogonal polarizations. An amplifier is disposed in circuit with the transmission line.

As is known in the art, reflect array antennas have been used in many applications. One type of reflect array antenna is a microstrip reflect array. The microstrip reflect antenna is essentially a planar array of microstrip patch antennas or dipoles illuminated by a feed. The individual antenna elements scatter the incident field appropriately so that the reflected field has a planar equi-phase front. The concept of a planar reflect array is not new, however, implementations found in the literature use a single antenna element for both transmit and receive. Pozar, et al., in a paper entitled "Design of a Millimeter Wave Microstrip Reflectarrays" published in IEEE Transactions on Antennas and Propagation, Vol. 45, No. 2, February 1997, for example, presented a microstrip reflect array of unique patch antennas, each sized for appropriate phasing, in which the same antenna element receives and transmits. With the exception that each antenna element is unique, the single substrate structure is comprised of rectangular patches on one side and a ground plane on the other. Bialkowski et al. have implemented a microstrip reflect array at X-band using aperture coupled patch antennas as reported in an article entitled "Design, Development, and Testing of X-Band Amplifying Reflectarrays" and published in IEEE Transactions on Antennas and Propogation, Vol. 50, August 2002. Isolation between transmit and receive have proven difficult with this approach since only one antenna is used with orthogonal slots for both transmit and receive. Further, U.S. Pat. No. 6,384,787 describes a flat reflectarray antenna.


A method of forming a thin film of a metal oxide on a substrate by coating the substrate with a solution comprising metal-organic precursors is disclosed. This method is applicable to, e.g., forming thin films of perovskite-phase titanates, zirconates, and/or niobates of divalent metals such as Ba, Sr, Pb and/or Ca. In one embodiment, a first precursor comprises a divalent metal coordinated to one or more organic ligands, and a second precursor comprises a tetravalent metal coordinated to one or more organic ligands are supplied in a common solution. A substrate 14 is coated with this solution (e.g. by spin coating) to form a preliminary thin film 10. Substrate heater 22 preferably heats substrate 14 to a temperature sufficient to react ligands from the first and second precursors in an ester elimination reaction which forms a volatile precursor 16. This reaction leaves an intermediate compound film 12 comprising the divalent metal and the tetravalent metal on the substrate. The substrate may be subsequently annealed to drive off unreacted ligands and/or fully crystallize the intermediate compound film into a perovskite-phase film 18.

Perovskite-phase mixed-metal oxide materials with the general formula ABO.sub.3 may possess one or more interesting and useful electrical properties, due to the existence of polar space groups within the perovskite-phase crystals. For example, perovskites may be ferroelectric materials, which have the ability to permanently retain an electric dipole after orientation in an electric field. Perovskites may also have a pyroelectric behavior, which is exhibited by a high dependence of dielectric constant on crystal temperature. Perovskites are often piezoelectric crystals, which exhibit a charge when mechanically stressed and mechanically deform when an electric field is applied. Finally, many perovskites may be operated in a linear electric field range where they exhibit extremely high dielectric constant (generally greater than 200). As such, perovskites find application, e.g., in DRAMs, capacitors, ferroelectric devices (e.g. FRAMs), infrared detectors, and piezoelectric transducers.
The formation of perovskite-phase mixed-metal oxides as thin films (i.e. less than 5 .mu.m in thickness) on the surface of a semiconductor substrate presents particular problems. To utilize such a film for one or more of the properties listed above, the film must generally be deposited with exacting stoichiometry, high homogeneity and purity, and a controlled grain size, or the desired property may be largely non-existent. Bulk ceramic manufacture of perovskites generally meets these constraints using high temperature (typically over 1000.degree. C.) processes which are generally unacceptable for the manufacture of microelectronic device thin films. Methods of forming some perovskites in thin film form from a solution of metal-organic precursors have been developed, although these typically require annealing temperatures in excess of 600.degree. C. in order to densify, remove impurities from, and/or fully crystallize a film. Perovskite thin film deposition processes which can be used at temperatures generally less than 500.degree. C. to deposit high quality films are desired for the manufacture of microelectronic devices.

Solution routes to metal oxide films through ester elimination reactions


In a target rotational angle determining routine, based on a roll moment and longitudinal acceleration that are calculated, target rotational angles of actuators on a subject side (one of front-wheel and rear-wheel sides) and a counterpart side are obtained using a common map. An actual rotational angle on the counterpart side is read. It is determined whether the absolute value of a difference obtained by subtracting the actual rotational angle supplied from the counterpart side from the target rotational angle on the counterpart side that is obtained by the subject side is equal to or greater than a set angle difference Δθ0. If YES, it is determined that roll stiffness on the counterpart side is insufficient, and the target rotational angle on the subject side is changed so that a roll stiffness distribution ratio between the front-wheel side and the rear-wheel side comes close to a set distribution ratio.

1. Field of the Invention
The invention relates to a suspension system of a vehicle, and more particularly to a suspension system which can control roll stiffness distribution between a front-wheel side and a rear-wheel side.
2. Description of Related Art
Each of Japanese Patent Application Publications No. JP-A-7-186674, No. JP-A-7-32845, and No. JP-A-5-319055 describes a suspension system which can control roll stiffness distribution. For example, the suspension system described in Japanese Patent Application Publication No. JP-A-7-186674 includes a front roll stiffness changing device; a rear roll stiffness changing device; a roll moment distribution estimating device; and a roll stiffness distribution control device. The front roll stiffness changing device changes roll stiffness on the front-wheel side of a vehicle. The rear roll stiffness changing device changes roll stiffness on the rear-wheel side. The roll moment distribution estimating device estimates distribution of a roll moment that is caused in the vehicle between the front-wheel side and the rear-wheel side of the vehicle. The roll stiffness distribution control device controls the front roll stiffness changing device and the rear roll stiffness changing device so as to distribute roll stiffness between the front-wheel side and the rear-wheel side according to the result of estimation made by the roll moment distribution estimating device.
In such a suspension system that can control roll stiffness distribution, if both of actuators of the front roll stiffness changing device and the rear roll stiffness changing device have sufficient driving performance, the roll stiffness distribution of the vehicle can be controlled as intended.
However, at least one of the actuators of the front roll stiffness changing device and the rear roll stiffness changing device may not have sufficient driving performance. For example, there may be the case where size of at least one of the actuators cannot be increased taking into account an installation space, a permissible weight, and the like. Also, there may be the case where the front roll stiffness changing device and rear roll stiffness changing device are the same, and therefore the driving performance of one of the actuators becomes insufficient. Also, the operation amount of the roll stiffness changing device may become insufficient due to insufficient response of the actuator.
For example, when the roll stiff changing device includes a stabilizer bar and an actuator that actively changes the torsion angle thereof, the stabilizer bar may be twisted by a roll moment before the stabilizer bar is twisted by an appropriate angle by the actuator, and the actuator may not be operated because great resistance torque has already been generated. If the speed at which the roll moment increases is low, the actuator can be operated by a predetermined operation amount. However, if the speed at which the roll moment increases is high, the actuator cannot be operated by the predetermined operation amount.
If the operation amount of the front roll stiffness changing device is insufficient, the roll stiffness on the rear-wheel side increases, and the vehicle steer characteristic shifts from a standard steer characteristic, which is a designed steer characteristic, to an oversteer characteristic (the standard steer characteristic is not necessarily a neutral steer characteristic). If the operation amount of the rear roll stiffness changing device is insufficient, the roll stiffness on the front-wheel side increases, and the vehicle steer characteristic shifts to an understeer characteristic.

Vehicle suspension system with roll stiffness distribution control


A headlight of a motor vehicle includes several light emitting devices which are arranged next to each other in at least one direction of the extent of the headlight. When the headlight is switched on, the light emitting devices along at least one direction of the headlight are switched on sequentially.

The present invention relates to a method for operating a headlight of a motor vehicle, wherein the headlight has a plurality of light emitting devices. The present invention also relates to a motor vehicle.
The following discussion of related art is provided to assist the reader in understanding the advantages of the invention, and is not to be construed as an admission that this related art is prior art to this invention.
The headlights for the low-beam headlight of motor vehicles have a predetermined light distribution for illuminating the road. In motor vehicles destined for right-hand traffic, the light distribution is curtailed on the left half of the motor vehicle by about 1%. This distribution is a compromise for the different classes of roads. The driver can activate the high-beam headlight in suitable traffic situations to further enhance the visibility at night. According to the Highway Code, the high-beam headlights must be deactivated in residential areas having regular road illumination. The high-beam headlight is hereby abruptly turned off and on without intermediate steps.
Modern motor vehicles increasingly use headlights with light emitting diodes (LEDs). The light emitting diodes are characterized by a long lifetime, high energy efficiency and enhanced design options. The LED headlights can be divided into several segments, with each segment typically including several light emitting diodes. The LED headlights can be used, for example, to provide a low-beam light function and a high-beam light function.
It would therefore be desirable and advantageous to obviate prior art shortcomings and to provide an improved method for operating a headlight of a motor vehicle wherein the headlight includes a plurality of light emitting devices.

Method for operating a headlight of a motor vehicle


A wheel lift system for efficiently lifting and positioning a wheel and tire onto a wheel hub. The wheel lift system includes a first arm and a second arm pivotally attached to one another, a plurality of caster wheels attached to the lower portion of the arms, and a first support and a second support extending from the arms respectively for supporting a wheel and tire. A bias member is preferably attached to the arms for providing a bias force that assists in elevating the wheel and tire. Alternatively, a bias unit may be attached to the arms for providing a bias force that assists in maintaining elevation of a wheel and tire.

1. Field of the Invention
The present invention relates generally to wheel lift devices and more specifically it relates to a wheel lift system for efficiently lifting and positioning a wheel and tire onto a wheel hub.
2. Description of the Related Art
Wheel lift devices have been in use for years. Conventional wheel lift devices are typically comprised of hydraulic/pneumatic bottle jack lifting system that lifts the wheel in position with respect to a wheel hub. Another method of lifting wheels into a desired position with respect to a wheel hub is by manually positioning the wheel with blocks and related objects. Further methods can be found using winches or electric motors that lift a wheel into position. Conventional wheel lift devices are relatively complex and expensive. In addition, conventional wheel lift devices are relatively cumbersome to operate.
While these devices may be suitable for the particular purpose to which they address, they are not as suitable for efficiently lifting and positioning a wheel and tire onto a wheel hub.
In these respects, the wheel lift system according to the present invention substantially departs from the conventional concepts and designs of the prior art, and in so doing provides an apparatus primarily developed for the purpose of efficiently lifting and positioning a wheel and tire onto a wheel hub.

This patent is subject to a terminal disclaimer.


A method includes obtaining a nonlinear process model modeling a nonlinear process to be controlled. The method also includes obtaining an objective function defining how the process is controlled. The method further includes obtaining a control model defining a dynamic feasible region associated with a controlled variable, where the controlled variable is associated with the process. In addition, the method includes controlling the nonlinear process by solving a control problem that includes the process model, control model, and objective function. The dynamic feasible region defined by the control model could represent a funnel region. The objective function could include terms for minimizing and optimizing adjustments to one or more manipulated variables associated with the process. Controlling the nonlinear process could include performing simultaneous control and optimization, where adjustments to the one or more manipulated variables are chosen to meet the control objectives and possibly to optimize and minimize the adjustments.

Processing facilities are often managed using process control systems. Example processing facilities include manufacturing plants, chemical plants, polymer plants, crude oil refineries, ore processing plants, and paper or pulp manufacturing and processing plants. Among other operations, process control systems typically manage the use of motors, valves, and other industrial equipment in the processing facilities.
In conventional process control systems, controllers are often used to control the operation of the industrial equipment in the processing facilities. The controllers could, for example, monitor the operation of the industrial equipment, provide control signals to the industrial equipment, and generate alarms when malfunctions are detected.
Model predictive control (MPC) technology is one type of control technology that has been developed and used in recent years. This type of control technology typically uses a linear model to represent a system to be controlled. A control algorithm uses the model and sensor data or other input data to determine how the system to be controlled is adjusted.

Apparatus and method for model predictive control (MPC) of a nonlinear process


A test apparatus or system for testing impact induced brain trauma a method of making and a method using the same are provided. The system includes a head model, which includes a skull component, a brain component, and a fluid component. The skull component has a wall defining an interior chamber. The brain component includes a gel material and is disposed within the interior chamber. The fluid component is disposed inside the interior chamber. The system may also include a fluid tank fluidly coupled with the skull component and configured to provide the fluid component into the interior chamber. The head model may further include a layer of porous media disposed between the brain component and the interior wall surface of the skull component. The system may include at least one impact element for providing an impact on the head model. The impact is translational or rotational or both.

Chronic traumatic encephalopathy (CTE) is a progressive degenerative disease resulting from a head trauma and particularly a history of repetitive head trauma. Military personnel may be exposed to blasts and other head impacts, which may lead to development of CTE. Other environments where people may be subjected to head trauma include the health care industry, and industrial environments such as in a factory or construction site. Athletes participating in contact sports such as football, soccer, rugby and boxing incur repetitive head trauma that has been shown to lead to the development of CTE in some individuals. CTE may result from symptomatic concussions as well as sub-concussive head trauma. Many athletes may experience frequency sub-concussive head trauma during participation in a contact sport and never have a symptomatic concussion. These athletes may still develop CTE. The effect of these frequent head impacts is a growing concern.
CTE may result from repetitive damage to axons in the brain, such as shearing caused by high acceleration of the brain tissue. High acceleration is caused by rapid head velocity change, such as that caused by an impact to the head. Axons connect neurons in the brain. Damage to the axons can result in immediate effects and/or delayed effects, such as CTE. Brain injury, such as axonal shearing, may create neurochemical and neurometabolic cascade effects. Even mild trauma to the brain can result in neuronal depolarization, which leads to neuronal discharge and the release of neurotransmitters and increased extra cellular potassium (K+). This may be followed by an increased glucose demand and metabolism (hyperglycolysis) and a resultant relative ischemia from reduced cerebral blood flow. Axonal injury may also result from an influx of extra cellular calcium that reduces cerebral blood flow through vasoconstriction, and the release of oxygen free radicals. These neurochemical and neurometabolic effects from even mild head trauma may result in the development of CTE.

Apparatus for examining brain injury, method of making and method of using the same