Method for identifying compounds useful for treating and/or preventing disease-associated bone loss

A method for identifying a compound which inhibits the activation of RAC GTPase by DOCK5 protein comprising the steps of:

• coexpressing at least the DHR2 domain of DOCK5 and the RAC proteins in a cell, wherein said at least the DHR2 domain of DOCK5 protein induces the conversion of inactive RAC, which inactive RAC is bound to GDP, to active RAC, which active RAC is bound to GTP,
• contacting or not said cell with said compound,
• determining the conversion of inactive RAC to active RAC in the presence or absence of said compound, and
• selecting the compound inhibiting the conversion of inactive RAC to active RAC.

A method for identifying a compound which inhibits the activation of RAC1/2 GTPase by inhibiting the binding of ELMO1 to the SH3 domain of DOCK5 comprising the steps of:

• a) contacting a test compound with the ELMO1 protein or a derivative thereof;
• b) determining the possible binding of said test compound to the ELMO1 protein or the derivative thereof;
• and optionally
• c) selecting the compound inhibiting the conversion of inactive RAC1/2 to active RAC1/2.

A method for identifying a compound which inhibits the activation of RAC1/2 GTPase by inhibiting the binding of ELMO1 to the SH3 domain of DOCK5 comprising the steps of:

• a) contacting a test compound with the ELMO1 protein or the derivative thereof and a polypeptide comprising at least the SH3 domain of DOCK5 or the derivative thereof;
• b) measuring the binding between said ELMO1 protein and said polypeptide in the presence or in the absence of said compound;
• and optionally
• c) selecting the compound inhibiting the conversion of inactive RAC1/2 to active RAC1/2.

Therapeutic method for treating and/or preventing bone loss diseases, comprising the administration of a therapeutically effective quantity of a compound selected from the group consisting of:

• 4-[5-(4-bromophenyl)-3-(4-nitrophenyl)-4,5-dihydro-1H-pyrazol-1-yl]-4-oxobutanoic acid;
• 2,2,2-trichloro-N-(1,1-dioxido-2,3-dihydro-3-thienyl)-N-(4-methylphenyl)acetamide;
• 3-(3-chlorophenyl)-7-methyl-4-methylene-3,4-dihydro-2(1H)-quinazolinone;
• 3-[4-(3-bromobenzylidene)-3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl]benzoic acid;
• N-2,1,3-benzothiadiazol-4-yl-5-bromo-2-furamide;
• 1-acetyl-4-(2-chloro-4-nitrophenyl)-2-methylpiperazine;
• 3-(3-methoxybenzylidene)-5-(4-methylphenyl)-2(3H)-furanone;
• 3-[5-(3,4-dichlorophenyl)-2-furyl]acrylic acid;
• (2-chloro-4-{[5-(2-chlorophenyl)-6-(ethoxycarbonyl)-7-methyl-3-oxo-5H-[1,3]thiazolo[3,2-a]pyrimidin-2(3H)-ylidene]methyl}-6-methoxyphenoxy)acetic acid;
• 4-{[4-(diphenylmethyl)-1-piperazinyl]sulfonyl}-2,1,3-benzothiadiazole;
• 4-[4-phenyl-5-(2-thienyl)-1H-imidazol-2-yl]-1,2-benzenediol;
• N-(3,4-dimethoxyphenyl)-4-[methyl(phenylsulfonyl)amino]benzamide;
• 1-[(2-hydroxyphenyl)carbonothioyl]-3-phenyl-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-ol;
• 2-methoxyethyl 4-[(4-tert-butylbenzoyl)amino]benzoate;
• N-(2,3-dichlorophenyl)-3-(5-methyl-2-furyl)acrylamide;
• N-(4-fluorophenyl)-3-[3-(trifluoromethyl)phenyl]acrylamide;
• 3-(2-furylmethyl)-2-(2-hydroxyphenyl)-2,3-dihydro-4(1H)-quinazolinone;
• N-(4-ethoxyphenyl)-2-{[5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl]thio}acetamide;
• 5-(4-nitrobenzylidene)-2-thioxo-3-[3-(trifluoromethyl)phenyl]-1,3-thiazolidin-4-one;
• (3,5-dichlorophenyl)[(phenylsulfonyl)carbonyl]amine;
• N-(2-bromophenyl)-3-(5-methyl-2-furyl)acrylamide;
• 2-(2-chlorophenoxy)-N-[2-chloro-5-(trifluoromethyl)phenyl]acetamide;
• N-[4-(4-acetyl-1-piperazinyl)phenyl]propanamide;
• 8-[(dimethylamino)methyl]-9-hydroxy-2-methyl-4H-pyrido[1,2-a]pyrimidin-4-one;
• 4-tert-butyl-N-[1-{[(2-methoxyphenyl)amino]carbonyl}-2-(2-thienyl)vinyl]benzamide;
• 2-chloro-N-(3-chloro-4-methoxyphenyl)benzamide;
• 2,6-di-tert-butyl-4-(2,3-dihydro-1H-perimidin-2-yl)phenol;
• 3-benzyl-2-(2,6-dichlorophenyl)-2,3-dihydro-4(1H)-quinazolinone;
• 1-(3,4-dichlorobenzyl)-1H-indole-3-carbaldehyde;
• N-[5-(1-adamantyl)-1,3,4-thiadiazol-2-yl]-N′-phenylurea;
• N-(3,4-dichlorophenyl)-N′-{5-[(4-methylphenoxy)methyl]-1,3,4-thiadiazol-2-yl}urea;
• N-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-(1-naphthyloxy)acetamide;
• N-[4-(4-acetyl-1-piperazinyl)phenyl]-4-ethoxy-3-nitrobenzamide;
• N-(2-chlorophenyl)-3-(4-fluorophenyl)acrylamide;
• 1-[(dimethyl-lambda˜4˜-sulfanylidene)amino]-2-methoxy-4-nitrobenzene;
• 5-benzylidene-1-(2-chlorophenyl)-2,4,6(1H,3H,5H)-pyrimidinetrione;
• 4-ethyl-5,6-dimethyl-2-phenylpyrimidine;
• 2-(3-chlorobenzylidene)-1H-indene-1,3(2H)-dione;
• 5-{5-[(3-methyl-5-oxo-1-phenyl-1,5-dihydro-4H-pyrazol-4-ylidene)methyl]-2-furyl}-1H-isoindole-1,3(2H)-dione;
• N-(2,5-dimethylphenyl)-3-(4-methoxyphenyl)acrylamide;
• 2-({2-[(4-nitrophenyl)amino]ethyl}amino)ethanol;
• N-(3-methoxyphenyl)-4-propoxybenzamide;
• 2-(4-hydroxyphenyl)-3-phenyl-2,3-dihydro-4(1H)-quinazolinone;
• 4-methyl-1-(2-nitrobenzoyl)piperidine;
• 2-hydroxy-N′-[(2-methylphenyl)sulfonyl]benzohydrazide;
• 4-(1,3-benzothiazol-2-yl)butanoic acid;
• 4-(3-methylbenzylidene)-1-phenyl-3,5-pyrazolidinedione;
• 4-(2,4-dichlorophenoxy)-N-(2-ethoxyphenyl)butanamide;
• N-(2-methoxyphenyl)-N′-(phenylsulfonyl)benzenecarboximidamide;
• N-[2-(2-chloro-5-iodophenyl)-1,3-benzoxazol-5-yl]-2-methylpropanamide;
• 5-(4-butoxyphenyl)-3-cyclohexyl-1,2,4-oxadiazole;
• N-(3,4-dichlorophenyl)-N′-4H-1,2,4-triazol-4-yl urea;
• 6-chloro-4-phenyl-3-[3-(3,4,5-trimethoxyphenyl)acryloyl]-2(1H)-quinolinone;
• 6-bromo-4-phenyl-3-[3-(3,4,5-trimethoxyphenyl)acryloyl]-2(1H)-quinolinone;
• and
• N-(1H-1,2,3-benzotriazol-1-ylmethyl)-4-nitro-1,2,5-oxadiazol-3-amine to a subject in need thereof.

Therapeutic method for treating and/or preventing bone loss diseases, comprising the administration of a therapeutically effective quantity of a pharmaceutical composition comprising at least one compound selected from the group consisting of:

• 4-[5-(4-bromophenyl)-3-(4-nitrophenyl)-4,5-dihydro-1H-pyrazol-1-yl]-4-oxobutanoic acid;
• 2,2,2-trichloro-N-(1,1-dioxido-2,3-dihydro-3-thienyl)-N-(4-methylphenyl)acetamide;
• 3-(3-chlorophenyl)-7-methyl-4-methylene-3,4-dihydro-2(1H)-quinazolinone;
• 3-[4-(3-bromobenzylidene)-3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl]benzoic acid;
• N-2,1,3-benzothiadiazol-4-yl-5-bromo-2-furamide;
• 1-acetyl-4-(2-chloro-4-nitrophenyl)-2-methylpiperazine;
• 3-(3-methoxybenzylidene)-5-(4-methylphenyl)-2(3H)-furanone;
• 3-[5-(3,4-dichlorophenyl)-2-furyl]acrylic acid;
• (2-chloro-4-{[5-(2-chlorophenyl)-6-(ethoxycarbonyl)-7-methyl-3-oxo-5H-[1,3]thiazolo[3,2-a]pyrimidin-2(3H)-ylidene]methyl}-6-methoxyphenoxy)acetic acid;
• 4-{[4-(diphenylmethyl)-1-piperazinyl]sulfonyl}-2,1,3-benzothiadiazole;
• 4-[4-phenyl-5-(2-thienyl)-1H-imidazol-2-yl]-1,2-benzenediol;
• N-(3,4-dimethoxyphenyl)-4-[methyl(phenylsulfonyl)amino]benzamide;
• 1-[(2-hydroxyphenyl)carbonothioyl]-3-phenyl-5-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-ol;
• 2-methoxyethyl 4-[(4-tert-butylbenzoyl)amino]benzoate;
• N-(2,3-dichlorophenyl)-3-(5-methyl-2-furyl)acrylamide;
• N-(4-fluorophenyl)-3-[3-(trifluoromethyl)phenyl]acrylamide;
• 3-(2-furylmethyl)-2-(2-hydroxyphenyl)-2,3-dihydro-4(1H)-quinazolinone;
• N-(4-ethoxyphenyl)-2-{[5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl]thio}acetamide;
• 5-(4-nitrobenzylidene)-2-thioxo-3-[3-(trifluoromethyl)phenyl]-1,3-thiazolidin-4-one;
• (3,5-dichlorophenyl) [(phenylsulfonyl)carbonyl]amine;
• N-(2-bromophenyl)-3-(5-methyl-2-furyl)acrylamide;
• 2-(2-chlorophenoxy)-N-[2-chloro-5-(trifluoromethyl)phenyl]acetamide;
• N-[4-(4-acetyl-1-piperazinyl)phenyl]propanamide;
• 8-[(dimethylamino)methyl]-9-hydroxy-2-methyl-4H-pyrido[1,2-a]pyrimidin-4-one;
• 4-tert-butyl-N-[1-{[(2-methoxyphenyl)amino]carbonyl}-2-(2-thienyl)vinyl]benzamide;
• 2-chloro-N-(3-chloro-4-methoxyphenyl)benzamide;
• 2,6-di-tert-butyl-4-(2,3-dihydro-1H-perimidin-2-yl)phenol;
• 3-benzyl-2-(2,6-dichlorophenyl)-2,3-dihydro-4(1H)-quinazolinone;
• 1-(3,4-dichlorobenzyl)-1H-indole-3-carbaldehyde;
• N-[5-(1-adamantyl)-1,3,4-thiadiazol-2-yl]-N′-phenylurea;
• N-(3,4-dichlorophenyl)-N′-{5-[(4-methylphenoxy)methyl]-1,3,4-thiadiazol-2-yl}urea;
• N-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-(1-naphthyloxy)acetamide;
• N-[4-(4-acetyl-1-piperazinyl)phenyl]-4-ethoxy-3-nitrobenzamide;
• N-(2-chlorophenyl)-3-(4-fluorophenyl)acrylamide;
• 1-[(dimethyl-lambda˜4˜-sulfanylidene)amino]-2-methoxy-4-nitrobenzene;
• 5-benzylidene-1-(2-chlorophenyl)-2,4,6(1H,3H,5H)-pyrimidinetrione;
• 4-ethyl-5,6-dimethyl-2-phenylpyrimidine;
• 2-(3-chlorobenzylidene)-1H-indene-1,3(2H)-dione;
• 5-{5-[(3-methyl-5-oxo-1-phenyl-1,5-dihydro-4H-pyrazol-4-ylidene)methyl]-2-furyl}-1H-isoindole-1,3(2H)-dione;
• N-(2,5-dimethylphenyl)-3-(4-methoxyphenyl)acrylamide;
• 2-({2-[(4-nitrophenyl)amino]ethyl}amino)ethanol;
• N-(3-methoxyphenyl)-4-propoxybenzamide;
• 2-(4-hydroxyphenyl)-3-phenyl-2,3-dihydro-4(1H)-quinazolinone;
• 4-methyl-1-(2-nitrobenzoyl)piperidine;
• 2-hydroxy-N′-[(2-methylphenyl)sulfonyl]benzohydrazide;
• 4-(1,3-benzothiazol-2-yl)butanoic acid;
• 4-(3-methylbenzylidene)-1-phenyl-3,5-pyrazolidinedione;
• 4-(2,4-dichlorophenoxy)-N-(2-ethoxyphenyl)butanamide;
• N-(2-methoxyphenyl)-N′-(phenylsulfonyl)benzenecarboximidamide;
• N-[2-(2-chloro-5-iodophenyl)-1,3-benzoxazol-5-yl]-2-methylpropanamide;
• 5-(4-butoxyphenyl)-3-cyclohexyl-1,2,4-oxadiazole;
• N-(3,4-dichlorophenyl)-N′-4H-1,2,4-triazol-4-yl urea;
• 6-chloro-4-phenyl-3-[3-(3,4,5-trimethoxyphenyl)acryloyl]-2(1H)-quinolinone;
• 6-bromo-4-phenyl-3-[3-(3,4,5-trimethoxyphenyl)acryloyl]-2(1H)-quinolinone;
• and
• N-(1H-1,2,3-benzotriazol-1-ylmethyl)-4-nitro-1,2,5-oxadiazol-3-amine
• and, optionally, a pharmaceutically acceptable support to a subject in need thereof.