Patent Application
20060035969
Copending application Ser. No. ______ [Attorney Docket No. 034227-592], filed concurrently herewith and also assigned to the assignee hereof.
1. Technical Field of the Invention
The present invention relates, as novel and useful industrial products, to a novel class of compounds which are modulators of the Peroxisome Proliferator-Activated Receptor (PPAR) type receptors. It also relates to their method of preparation and to their formulation into pharmaceutical compositions for administration in human or veterinary medicine, or alternatively in cosmetic compositions.
2. Description of Background and/or Related and/or Prior Art
The activity of the PPAR-type receptors has been the subject of numerous studies. There may be mentioned, as a guide, the publication entitled “Differential Expression of Peroxisome Proliferator-Activated Receptor Subtypes During the Differentiation of Human Keratinocytes”, Michel Rivier et al., J. Invest. Dermatol., 111, 1998, p. 1116-1121, in which a large number of bibliographic references relating to PPAR-type receptors is listed. There may also be mentioned, as a guide, the dossier entitled “The PPARs: From orphan receptors to Drug Discovery”, Timothy M. Willson, Peter J. Brown, Daniel D. Sternbach, and Brad R. Henke, J. Med. Chem., 2000, Vol. 43, p. 527-550.
The PPAR receptors activate transcription by binding to elements of DNA sequences, called peroxisome proliferator response elements (PPRE), in the form of a heterodimer with the retinoid X receptors (known as RXRs).
Three human PPAR subtypes have been identified and described: PPARα, PPARγ and PPARδ (or NUC1).
PPARα is mainly expressed in the liver while PPARδ is ubiquitous.
It is described in WO 98/32444 that PPARα selective compounds play a role in the barrier function and the differentiation of the stratum corneum.
PPARγ is the most widely studied of the three subtypes. All the references suggest a critical role of the PPARγ receptors in the regulation of differentiation of adipocytes, where it is highly expressed. It also plays a key role in systemic lipid homeostasis.
It has in particular been described in WO 96/33724 that PPARγ-selective compounds, such as prostaglandin-J2 or -D2, are potential active agents for treating obesity and diabetes.
A novel class of PPAR-modulating compounds has now been developed.
Thus, the present invention features novel compounds having the following general formula (I):
in which:
In particular, when the compounds according to the invention are provided in the form of salts, they are salts of an alkali or alkaline-earth metal, zinc salts, or salts of an organic amine.
According to the present invention, the expression “hydroxyl radical” means the —OH radical.
According to the present invention, the expression “alkyl radical” having from 1 to 12 carbon atoms means a hydrogenated or fluorinated, linear or cyclic, optionally branched, radical containing 1 to 12 carbon atoms which may be interrupted by one or more heteroatoms, and preferably the alkyl radicals having from 1 to 12 carbon atoms are methyl, ethyl, isopropyl, butyl, tert-butyl, hexyl, octyl, decyl or cyclohexyl radicals.
The expression “monohydroxyalkyl radical” means a radical having 1 to 6 carbon atoms, and preferably having from 2 to 3 carbon atoms, in particular a 2-hydroxyethyl, 2-hydroxypropyl or 3-hydroxypropyl radical.
The expression “polyhydroxyalkyl radical” means a radical containing from 3 to 6 carbon atoms and from 2 to 5 hydroxyl groups, such as 2,3-dihydroxypropyl, 2,3,4-trihydroxybutyl or 2,3,4,5-tetrahydroxypentyl radicals, or the pentaerythritol residue.
The expression “polyether radical” means a polyether radical having from 1 to 6 carbon atoms interrupted by at least one oxygen atom such as methoxymethoxy, ethoxymethoxy or methoxyethoxymethoxy radicals.
The expression “alkoxy radical having from 1 to 7 carbon atoms” means a radical containing from one to seven carbon atoms such as the methoxy, ethoxy, isopropyloxy, tert-butoxy, hexyloxy, benzyloxy or phenoxy radicals, which may be optionally substituted with an alkyl radical having from 1 to 12 carbon atoms.
The expression “aryl radical” means a phenyl, biphenyl, cinnamyl or naphthyl radical which may be mono- or disubstituted with a halogen atom, a radical CF3, an alkyl radical having from 1 to 12 carbon atoms, an alkoxy radical having from 1 to 7 carbon atoms, a nitro functional group, a polyether radical, an aryl radical, a benzoyl radical, an alkyl ester group, a carboxylic acid, a hydroxyl radical optionally protected by an acetyl or benzoyl group or an amino functional group optionally protected by an acetyl or benzoyl group or optionally substituted with at least one alkyl having from 1 to 12 carbon atoms.
The expression “aralkyl radical” means a benzyl, phenethyl or naphthalen-2-ylmethyl radical which may be mono- or disubstituted with a halogen atom, a radical CF3, an alkyl radical having from 1 to 12 carbon atoms, an alkoxy radical having from 1 to 7 carbon atoms, a nitro functional group, a polyether radical, an aryl radical, a benzoyl radical, an alkyl ester group, a carboxylic acid, a hydroxyl radical optionally protected by an acetyl or benzoyl group or an amino functional group optionally protected by an acetyl or benzoyl group or optionally substituted with at least one alkyl having from 1 to 12 carbon atoms.
The expression “heteroaryl radical” is preferably understood to mean an aryl radical interrupted by one or more heteroatoms, such as the pyridyl, furyl, thienyl, isoxazolyl, oxadiazolyl, oxazolyl, isothiazolyl, quinazolinyl, benzothiadiazolyl, benzimidazole, indolyl or benzofuran radical, optionally substituted with at least one halogen, an alkyl having from 1 to 12 carbon atoms, an alkoxy having from 1 to 7 carbon atoms, an aryl radical, a nitro functional group, a polyether radical, a heteroaryl radical, a benzoyl radical, an alkyl ester group, a carboxylic acid, a hydroxyl optionally protected by an acetyl or benzoyl group or an amino functional group optionally protected by an acetyl or benzoyl group or optionally substituted with at least one alkyl having from 1 to 12 carbon atoms.
The expression “heterocycle” is preferably understood to mean the morpholino, piperidino, piperazino, 2-oxopiperidin-1-yl and 2-oxopyrrolidin-1-yl radicals optionally substituted with at least one alkyl group having from 1 to 12 carbon atoms, an alkoxy having from 1 to 7 carbon atoms, an aryl radical, a nitro functional group, a polyether radical, a heteroaryl radical, a benzoyl radical, an alkyl ester group, a carboxylic acid, a hydroxyl optionally protected by an acetyl or benzoyl group or an amino functional group optionally protected by an acetyl or benzoyl group or optionally substituted with at least one alkyl having from 1 to 12 carbon atoms.
Among the compounds of formula (I) falling within the scope of the present invention, the following compounds may be mentioned in particular (alone or as a mixture):
A general description of the preparation of the compounds of general formula of the appended Figure of Drawing is given below.
The reaction scheme described in Figure of Drawing is a general scheme allowing the production of the compounds according to the invention.
The compounds of general formula (I) may be obtained (Figure of Drawing) by coupling a thiol, an alcohol, an amine or a seleniated derivate (depend on X value) with an aromatic iodinated compound, using a metal catalyst such as nickel or palladium derivatives, in the presence of a hydride donor such as sodium borohydride and if necessary a base. Concerning diaryl amine compounds, the copper or palladium catalyzed amination (Tetrahedron 58, (2002) 2041-2075) of the nitro aniline compound with aryl halogenide may be employed, followed by the reduction of the nitro to the corresponding amino group. Concerning the preparation of diaryl ether coupling of the corresponding alkoxide catalyzed by palladium may be employed. Concerning the preparation of diaryl ketone compounds, palladium catalyzed conversion of halogenoaryl derivatives compound to the corresponding organotin derivatives followed by a palladium catalyzed coupling with acyl chloride derivative may afford the target product. The ketone might be protected in order to avoid problems during reductive amination. The next step is a reductive amination of the preceding amine and of an aldehyde, which may be carried out with isolation of the intermediate imine or otherwise, followed by reduction of the latter by the action of a reducing agent such as NaBH3CN. The alkylated amine obtained can then be subjected to the action of an isocyanate or an isothiocyanate in a solvent such as dichloromethane to give the corresponding urea or thiourea. It can also be further alkylated by reductive amination reaction in the presence of an aldehyde under the same conditions as above. The amide may also be formed by the action of an acid in the presence of a coupling agent such as O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) in the presence of a base such as DIEA or an acyl halide and a base. The derivatives obtained are then saponified by the action, for example, of a base such as NaOH to give the corresponding acids. The sulfated compounds (X: S) oxydated by the action of metachloroperbenzoic acid (MCPBA) in the presence of dichloromethane.
The compounds according to the invention have PPAR-type receptor modulating properties. This activity on the PPARα, δ and γ receptors is measured in a transactivation test and quantified by the dissociation constant Kdapp (apparent), as described in Example 51.
The preferred compounds of the present invention have a dissociation constant of less than or equal to 1,000 nM, and advantageously of less than or equal to 500 nM for at least one of the PPAR subtypes.
The present invention also features medicaments containing the compounds of formula (I) as described above.
The present invention also features formulating the compounds of formula (I) into compositions suited for regulating and/or restoring the metabolism of skin lipids.
The compounds according to the invention are particularly suitable in the fields of the following treatments:
The present invention also features pharmaceutical compositions comprising, formulated into a physiologically acceptable medium, at least one compound of formula (I) as defined above.
The administration (regime or regimen) of the compositions according to the invention may be carried out enterally, parenterally, topically or ocularly. Preferably, the pharmaceutical composition is packaged in a form suitable for application by the topical route.
By the enteral route, the composition may be provided in the form of tablets, gelatin capsules, sugar-coated tablets, syrups, suspensions, solutions, powders, granules, emulsions, suspensions of lipid or polymeric microspheres or nanospheres or vesicles allowing controlled release. By the parenteral route, the composition may be provided in the form of solutions or suspensions for perfusion or injection.
The compounds according to the invention are generally administered at a daily dose of about 0.001 mg/kg to 100 mg/kg of body weight, in 1 to 3 doses.
The compounds are administered by the systemic route at a concentration generally of from 0.001% to 10% by weight, preferably from 0.01% to 1% by weight, relative to the weight of the composition.
By the topical route, the pharmaceutical compositions according to the invention are more particularly suited for the treatment of the skin and the mucous membranes and may be provided in the form of salves, creams, milks, ointments, powders, impregnated pads, syndets, solutions, gels, sprays, mousses, suspensions, lotions, sticks, shampoos or washing bases. They may also be provided in the form of suspensions of lipid or polymeric microspheres or nanospheres or vesicles or of polymeric patches and of hydrogels allowing controlled release. This composition for the topical route may be provided in anhydrous form, in aqueous form or in the form of an emulsion.
The compounds are administered by the topical route at a concentration which is generally from 0.001% to 10% by weight, preferably from 0.01% to 1% by weight, relative to the total weight of the composition.
The compounds of formula (I) according to the invention also find application in the cosmetics field, in particular in body and hair care, and more particularly for regulating and/or restoring skin lipid metabolism.
This invention therefore also features the cosmetic application of a composition comprising, in a physiologically acceptable carrier, at least one of the compounds of formula (I) for body or hair care.
The cosmetic compositions according to the invention containing, in a cosmetically acceptable carrier, at least one compound of formula (I) or one of its optical or geometric isomers or one of its salts, may be provided in particular in the form of a cream, a milk, a lotion, a gel, suspensions of lipid or polymeric microspheres or nanospheres or vesicles, impregnated pads, solutions, sprays, mousses, sticks, soaps, shampoos or washing bases.
The concentration of compound of formula (I) in the cosmetic composition is preferably from 0.001% to 3% by weight, relative to the total weight of the composition.
The pharmaceutical and cosmetic compositions as described above may in addition contain inert additives, or even pharmacodynamically active additives as regards the pharmaceutical compositions, or combinations of these additives, and in particular:
Of course, one skilled in the art will be careful to choose the possible compound(s) to be added to these compositions such that the advantageous properties intrinsically associated with the present invention are not or not substantially impaired by the addition envisaged.
In order to further illustrate the present invention and the advantages thereof, the following specific examples are given, including those relating to the preparation of the compounds (I) as well as the biological activity and particular formulations thereof, it being understood that same are intended only as illustrative and in nowise limitative. In said examples to follow, all parts and percentages are given by weight, unless otherwise indicated.
The products were analyzed by HPLC/Mass. Column: 2.1×5 mm, 3μ, High purity C18 Hypersil.
Mobile phase: A (CH3CN/0.1 v/v HCO2H); B (H2O/0.1 v/v HCO2H),
Waters Alliance 2790 LC Mobile Phase
Solvents
The gradient contains 3 entries which are:
6.5 ml (0.12 mol) of concentrated sulfuric acid are added dropwise over a mixture of 10 g (0.06 mol) of 3-mercaptophenylacetic acid in 200 ml of ethanol. The reaction medium is then heated under reflux for 4 h. 1 ml of concentrated sulfuric acid is added dropwise in order to complete the reaction. The reaction medium is heated for 3 h under reflux and then concentrated in a rotary evaporator under vacuum. Water is added to the residue obtained. The solution is neutralized by adding sodium bicarbonate. The desired product is extracted by adding ethyl ether. The organic phase is washed with water, dried over magnesium sulfate and concentrated in a rotary evaporator. The product is purified by chromatography on a silica column, eluted with dichloromethane. After evaporation of the solvents, 10.15 g (86%) of the expected compound are recovered in the form of a yellow oil.
A solution of ethyl 3-mercaptophenylacetate (2.5 g, 12.7 mmol) in 10 ml of THF is added to a mixture of borohydride polymer supported Amberlite® IRA400 resin (2.5 mmol/g) (Aldrich: 32864-2) (10.1 g, 25.4 mmol), bis(bipyridine)nickel (II) bromide (105.3 mg) (Organometallics 1985, 4, 657-661) and 4-iodoaniline (1.8 g, 8.5 mmol) in ethanol (90 ml). The mixture is stirred under reflux for 3 h. The reaction medium is filtered and the filtrate concentrated in a rotary evaporator under vacuum. The product is purified by chromatography on a silica column (dichloromethane). After evaporation of the solvents, the expected product 524 mg (73%) is isolated in the form of a yellow oil.
1H NMR (CDCl3, 400 MHz): 1.22 (3H, t), 3.50 (2H, s), 3.65 (2H, NH2, s), 4.11 (2H, q), 6.65 (2H, Ar, d), 6.99 (2H, Ar, t), 7.04 (1H, Ar, s), 7.14 (1H, Ar, t), 7.28 (2H, Ar, d).
A mixture of product 1(b) (50 mg, 0.174 mmol), sodium hydroxide (50 mg), water (50 μl) and ethanol (50 μl) in THF (2.5 ml) is stirred for 24 h at room temperature. The reaction medium is acidified by adding a 2N hydrochloric acid solution and extracted with ethyl ether. The organic phase is washed with water, dried over magnesium sulfate and concentrated in a rotary evaporator under vacuum. The product is obtained in the form of a yellow solid (44 mg), 98%.
In a manner similar to Example 1(b), by reacting ethyl 3-mercaptophenylacetate (3 g, 15.3 mmol), 10 ml of THF, borohydride polymer supported Amberlite® IRA400 resin (2.5 mmol/g) (Aldrich: 32864-2) (12.24 g, 30.6 mmol), bis(bipyridine)nickel (II) bromide (127 mg) (Organometallics 1985, 4, 657-661) and 3-iodoaniline (2.2 g, 10.2 mmol), 2.38 g (54%) of the expected derivative are obtained in the form of a yellow oil.
1H NMR (CDCl3, 400 MHz): 1.22 (3H, t), 3.54 (4H, s), 4.12 (2H, q), 6.50 (1H, Ar, d), 6.653 (1H, Ar, s), 6.71 (1H, Ar, d), 7.05 (1H, Ar, t), 7.10 to 7.20 (1H, Ar, m), 7.22 (2H, Ar, d) 7.27 (1H, Ar, s).
In a manner similar to Example 1(c), by reacting ethyl [3-(3-aminophenylphenylsulfanyl)phenyl]acetate (50 mg, 0.174 mmol), sodium hydroxide (50 mg), water (50 μl) and ethanol (50 μl) in THF (2.5 ml), 39 mg (87%) of the expected derivative are obtained in the form of a yellow solid.
A solution of phenylacetaldehyde (163 mg, 1.357 mmol) and acetic acid (1 ml) is added to a solution of ethyl [3-(3-aminophenylsulfanyl)phenyl]-acetate (Example 2(a)) (390, 1.36 mmol) in 15 ml of DMF. 170.54 mg of sodium cyanoborohydride (2.71 mmol) are added and the mixture is stirred for 12 h at room temperature. After extracting with ethyl ether, the organic phase is washed with water, dried over magnesium sulfate and concentrated in a rotary evaporator under vacuum. The product is purified by chromatography on a silica column (dichloromethane 7/heptane 3). After evaporation of the solvents, the expected compound 170 mg (34%) is isolated in the form of a yellow oil.
1H NMR (CDCl3, 400 MHz): 1.21 (3H, t), 2.85 (2H, t), 3.32 (2H, t), 3.53 (2H, s), 4.09 (2H, q), 6.46 (1H, Ar, d), 6.60 (1H, Ar, s), 6.66 (1H, Ar, d), 7.06 to 7.29 (10H, Ar, m).
A mixture of ethyl [3-(3-phenethylaminophenylsulfanyl)phenyl]acetate (47 mg), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml) is stirred at room temperature for 4 days. After acidifying with 5 ml of a 1N hydrochloric acid solution, extracting with ethyl ether, the organic phase is washed with water, dried over magnesium sulfate and concentrated in a rotary evaporator under vacuum. After evaporation of the solvent, the expected compound 40 mg (92%) is isolated in the form of a yellow oil.
In a manner similar to Example 3(a), by reacting 3-phenylpropionaldehyde (182 mg, 1.357 mmol), acetic acid (1 ml), ethyl [3-(3-aminophenylsulfanyl)phenyl]acetate (Example 2(a)) (390, 1.36 mmol) in 15 ml of DMF, and 170.54 mg of sodium cyanoborohydride (2.71 mmol), 364 mg (66%) of the expected derivative are obtained in the form of a colorless oil.
1H NMR (CDCl3, 400 MHz): 1.19 (3H, t), 1.18 to 1.88 (2H, m), 2.64 (2H, t), 3.03 (2H, t), 3.50 (2H, s), 4.09 (2H, q), 6.39 (1H, Ar, d), 6.54 (1H, Ar, s), 6.64 (1H, Ar, d), 7.04 to 7.26 (10H, Ar, m).
In a manner similar to Example 3(b), by reacting ethyl {3-[3-(3-phenylpropylamino)phenylsulfanyl]phenyl}acetate (25 mg, 0.062 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 17 mg (73%) of the expected derivative are obtained in the form of a yellow oil.
In a manner similar to Example 3(a), by reacting heptaldehyde (155 mg, 1.357 mmol), acetic acid (1 ml), ethyl [3-(3-aminophenylsulfanyl)phenyl]acetate (Example 2(a)) (390, 1.36 mmol) in 15 ml of DMF, and 170.54 mg of sodium cyanoborohydride (2.71 mmol), 217 mg (42%) of the expected derivative are obtained in the form of a colorless oil.
1H NMR (CDCl3, 400 MHz): 0.88 (3H, t), 1.22 (3H, t), 1.27 to 1.31 (8H, m), 1.52 to 1.59 (2H, m), 3.03 (2H, t), 3.44 (1H, NH, s), 3.54 (2H, s), 4.12 (2H, q), 6.45 (1H, Ar, d), 6.58 (1H, Ar, s), 6.65 (1H, Ar, d), 7.07 (1H, Ar, t), 7.11 (1H, Ar, t), 7.21 (2H, Ar, d), 7.26 (1H, Ar, s).
In a manner similar to Example 3(b), by reacting ethyl [3-(3-heptylaminophenylsulfanyl)phenyl]acetate (40 mg, 0.10 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 34 mg (93%) of the expected derivative are obtained in the form of a yellow oil.
In a manner similar to Example 3(a), by reacting butyraldehyde (97.9 mg, 1.357 mmol), acetic acid (1 ml), ethyl [3-(3-aminophenylsulfanyl)phenyl]acetate (Example 2(a)) (390, 1.36 mmol) in 15 ml of DMF, and 170.54 mg of sodium cyanoborohydride (2.71 mmol), 319 mg (69%) of the expected derivative are obtained in the form of a colorless oil.
1H NMR (CDCl3, 400 MHz): 0.91 (3H, t), 1.21 (3H, t), 1.32 to 1.42 (2H, m), 1.49 to 1.56 (2H, m), 3.02 (2H, t), 3.52 (3H, s), 4.11 (2H, q), 6.44 (1H, Ar, d), 6.57 (1H, Ar, s), 6.64 (1H, Ar, d), 7.06 (1H, Ar, t), 7.11 (1H, Ar, t), 7.18 (2H, Ar, d), 7.26 (1H, Ar, s).
In a manner similar to Example 3(b), by reacting ethyl [3-(3-butylaminophenylsulfanyl)phenyl]acetate (30 mg, 0.087 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 19 mg (89%) of the expected derivative are obtained in the form of a yellow oil.
In a manner similar to Example 3(a), by reacting heptaldehyde (160.5 mg, 1.15 mmol), acetic acid (1 ml), ethyl [3-(4-aminophenylsulfanyl)phenyl]acetate (Example 1b) (330 mg, 1.15 mmol) in 15 ml of DMF, and 144.5 mg of sodium cyanoborohydride (2.3 mmol) and adding 160.5 μl of heptaldehyde after stirring for 4 h, 343 mg (77%) of the expected derivative are obtained in the form of a colorless oil.
1H NMR (CDCl3, 400 MHz): 0.88 (3H, t), 1.22 (3H, t), 1.28 to 1.41 (6H, m), 1.58 to 1.63 (2H, m), 3.10 (2H, t), 3.50 (2H, s), 4.11 (2H, q), 6.58 (2H, Ar, d), 6.98 (2H, Ar, t), 7.03 (1H, Ar, s), 7.13 (1H, Ar, t), 7.32 (2H, Ar, d).
In a manner similar to Example 3(b), by reacting ethyl [3-(4-heptylaminophenylsulfanyl)phenyl]acetate (45 mg, 0.12 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 30 mg (72%) of the expected derivative are obtained in the form of a yellow oil.
In a manner similar to Example 3(a), by reacting butyraldehyde (103.6 mg, 1.15 mmol), acetic acid (1 ml), ethyl [3-(4-aminophenylsulfanyl)phenyl]acetate (Example 1b) (330 mg, 1.15 mmol) in 15 ml of DMF, and 144.5 mg of sodium cyanoborohydride (2.71 mmol), 223 mg (66%) of the expected derivative are obtained in the form of a colorless oil.
1H NMR (CDCl3, 400 MHz): 0.95 (3H, t), 1.21 (3H, t), 1.37 to 1.46 (2H, m), 1.55 to 1.63 (2H, m), 3.10 (2H, t), 3.49 (2H, s), 3.76 (1H, NH, s), 4.105 (2H, q), 6.56 (2H, Ar, d), 6.95 (2H, Ar, t), 7.03 (1H, Ar, s), 7.12 (1H, Ar, t), 7.30 (2H, Ar, d).
In a manner similar to Example 3(b), by reacting ethyl [3-(4-butylaminophenylsulfanyl)phenyl]acetate (20 mg, 0.06 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 19 mg (100%) of the expected derivative are obtained in the form of a yellow oil.
In a manner similar to Example 3(a), by reacting phenylacetaldehyde (115 mg, 1.15 mmol), acetic acid (1 ml), ethyl [3-(4-aminophenylsulfanyl)phenyl]acetate (Example 1b) (330 mg, 1.15 mmol) in 15 ml of DMF, and 120.15 mg of sodium cyanoborohydride (2.3 mmol) and adding 33.5 μl of phenylacetaldehyde, after stirring for 4 h, 503 mg of the expected derivative and of a residual amine are obtained. The mixture is solubilized in 15 ml of DMF, and 400 mg of PS-benzaldehyde resin, acetic acid (1 ml) and sodium cyanoborohydride (58 mg) are added. The solution is stirred for 48 h at room temperature. After extracting with ethyl ether, the organic phase is washed with water, dried over magnesium sulfate and concentrated in a rotary evaporator under vacuum. The product is purified by filtration on a silica column (dichloromethane 8/heptane 2). After evaporation of the solvents, the expected compound 276 mg (34%) is isolated in the form of a yellow oil.
1H NMR (CDCl3, 400 MHz): 1.18 (3H, t), 2.86 (2H, t), 3.55 (2H, t), 3.47 (2H, s), 4.08 (2H, q), 6.53 (2H, Ar, d), 6.96 (2H, Ar, t), 7.03 (1H, Ar, s), 7.11 (1H, Ar, t), 7.17 to 7.22 (3H, Ar, m), 7.39 (4H, Ar, t).
In a manner similar to Example 3(b), by reacting ethyl [3-(4-phenethylaminophenylsulfanyl)phenyl]acetate (24 mg, 0.06 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 18 mg (81%) of the expected derivative are obtained in the form of a yellow oil.
A suspension of molecular sieve (1 g), 3-phenylpropionaldehyde (280 mg, 2.1 mmol), in THF (5 ml) is added to a solution of ethyl [3-(4-aminophenylsulfanyl)phenyl]acetate (Example 1b) (500 mg, 1.74 mmol) in 20 ml of ethanol and 10 ml of THF. The mixture is stirred for 24 h at room temperature. After filtering, 98.7 mg of sodium borohydride (2.71 mmol) are added to the filtrate. After stirring for 24 h and extracting with ethyl ether, the organic phase is washed with water, dried over magnesium sulfate and concentrated in a rotary evaporator under vacuum. The product is purified by chromatography on a silica column (dichloromethane 7/heptane 3). After evaporation of the solvents, the expected compound is isolated in the form of a yellow oil.
1H NMR (CDCl3, 400 MHz): 1.21 (3H, t), 1.91 to 1.98 (2H, m), 2.72 (2H, t), 3.14 (2H, t), 3.49 (2H, s), 4.06 to 4.13 (2H, m), 6.527 (2H, Ar, d), 7.00 (2H, Ar, t), 7.03 (1H, Ar, s), 7.13 (1H, Ar, t), 7.18 to 7.29 (3H, Ar, m), 7.27 to 7.31 (4H, Ar, m).
In a manner similar to Example 3(b), by reacting ethyl {3-[4-(3-phenylpropylamino)phenylsulfanyl]phenyl}acetate (30 mg, 0.07 mmol), a 35% sodium hydroxide solution (100 μl) and 50 μl of ethanol in THF (2 ml), 22 mg (96%) of the expected derivative are obtained in the form of a yellow oil.
Examples 11 to 20 were obtained by parallel chemistry. The reactions of a starting amine and a starting isocyanate are performed in several reactors simultaneously according to the operating protocol described below.
Operating Protocol:
0.123 mmol (50 mg) of ethyl {3-[4-(3-phenylpropylamino)phenylsulfanyl]phenyl}acetate (compound of Example 10a) is introduced into each 5 ml reactor. 2 ml of dichloromethane are added. Next, 0.247 mmol of isocyanate (see Table 2) are added. The reactors are stirred for 7 h at room temperature. 0.247 mmol of isocyanates is added if the starting amine has not completely disappeared (TLC monitoring). In this case, the stirring is continued for 12 h at room temperature.
The reaction media are concentrated to dryness for 2 h at 40° C. in a centrifugal evaporator under vacuum. The products are purified by filtration on silica cartridges (6 ml), 1: DCM, 2: DCM 80/AcOEt 20, and then concentrated to dryness, 2 h at 40° C. in a centrifugal evaporator (see Table 3 for the quantities obtained).
Operating Protocol:
0.128 mmol (50 mg) of ethyl [3-(4-phenethylaminophenylsulfanyl)phenyl]acetate (compound of Example 9a) is introduced into each 5 ml reactor. 2 ml of dichloromethane is added. Next, 0.255 mmol of isocyanate (see Table 4) is added. The reactors are stirred for 7 h at room temperature. 0.255 mmol of isocyanates is added if the starting amine has not completely disappeared (TLC monitoring). In this case, the stirring is continued for 12 h at room temperature.
The reaction media are concentrated to dryness for 2 h at 40° C. in a centrifugal evaporator under vacuum. The products are purified by filtration on silica cartridges (6 ml), 1: DCM, 2: DCM 80/AcOEt 20, and then concentrated to dryness, 2 h at 40° C. in a centrifugal evaporator (see Table 5 for the quantities obtained).
The esters obtained above (Examples 1a to 20a of Tables 3 and 5) are solubilized in 2 ml of THF. 100 μl of ethanol are then introduced. 100 μl of a sodium hydroxide solution at 35% are then added. The mixture is stirred at room temperature for 48 hours. The progress of the reaction is monitored by thin-layer chromatography (DCM 80/AcOEt 20). After extracting with ether, acidifying with a 1N hydrochloric acid solution, the organic phase is washed twice with water, dried over magnesium sulfate and concentrated to dryness in a centrifugal evaporator under vacuum. The products are purified by filtration on silica cartridges (6 ml) if necessary, and then concentrated to dryness for 2 h at 40° C. in a centrifugal evaporator under vacuum. The final products are analyzed by mass-coupled HPLC (Table 6).
Examples 21 to 50 were obtained by parallel chemistry. The reactions of a starting amine and a starting isocyanate are performed in several reactors simultaneously according to the operating protocol described below.
Operating Protocol:
0.077 mmol of amine (see Table 7) is introduced into each 5 ml reactor. 2 ml of dichloromethane are added. Next, 0.153 mmol of isocyanate (see Table 8) is added. The reactors are stirred for 7 h at room temperature. 0.062 mmol of isocyanates is added if the starting amine has not completely disappeared (TLC monitoring). In this case, the stirring is continued for 12 h at room temperature.
The reaction media are concentrated to dryness for 2 h at 40° C. in a centrifugal evaporator under vacuum. The products are purified by filtration on silica cartridges (6 ml), 1: DCM, 2: DCM 80/AcOEt 20, and then concentrated to dryness, 2 h at 40° C. in a centrifugal evaporator.
Each of the esters obtained above is solubilized in 2 ml of THF. 100 μl of ethanol are then introduced. 100 μl of a sodium hydroxide solution at 35% are then added. The mixture is stirred at room temperature for 48 h. The progress of the reaction is monitored by thin-layer chromatography (DCM 80/AcOEt 20). After extracting with ether, acidifying with a 1N hydrochloric acid solution, the organic phase is washed twice with water, dried over magnesium sulfate and concentrated to dryness in a centrifugal evaporator under vacuum. The products are purified by filtration on silica cartridges (6 ml) if necessary, and then concentrated to dryness for 2 h at 40° C. in a centrifugal evaporator under vacuum. The final products are analyzed by mass-coupled HPLC (Table 9).
Compounds 21a to 50a are the esters corresponding to the acids 21b to 50b before the saponification reaction.
Amidation:
23 mg (52.3 μmol) of {3-[4-(1-butyl-3-cyclohexylureido)phenylsulfanyl]phenyl}acetic acid obtained above in Example 34b are dissolved in 0.5 ml of DMF. 76 mg of carbodiimide supported on polystyrene (105 μmol) and 30 mg (78.4 μmol) of HATU (N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate) are introduced into the solution. The mixture is stirred for 15 minutes at room temperature. A solution of 52.3 μmol of the desired amine in 0.5 ml of dichloromethane is then added.
The reaction medium is stirred for 4 h at room temperature and then filtered and concentrated to dryness in a centrifugal evaporator under vacuum. The products are purified by filtration on silica cartridges (2 g) (eluent: heptane/AcOEt 1/1), and then concentrated to dryness at 40° C. in a centrifugal evaporator under vacuum. The final products are analyzed by mass-coupled HPLC.
A mixture of 239 mg (697 μmol) of ethyl [3-(4-butylaminophenylsulfanyl)phenyl]acetate (Example 8a), triethylamine (211 mg, 2.09 mmol) and cyclohexyl isocyanate (174 mg, 1.39 mmol) in 2 ml of dichloromethane is stirred for 8 h at room temperature. 174 mg of cyclohexyl isocyanate and 211 mg of triethylamine are added to the mixture and the stirring is continued for 14 h at room temperature. 0.3 ml of cyclohexyl isocyanate is added and the mixture is heated at 40° C. for 3 h. The reaction medium is concentrated to dryness in a centrifugal evaporator under vacuum. The expected product is purified by filtration on silica cartridges (8 g) (eluent: heptane/AcOEt 20/5), and then concentrated to dryness at 40° C. in a centrifugal evaporator under vacuum. The final product is obtained in the form of a colorless oil and analyzed by mass-coupled HPLC. HPLC (% total of the surface area): 100%, ES MASS (M+H+): 469.2.
1H NMR (CDCl3, 400 MHz): 0.99 (3H, t), 1.25 to 1.33 (9H, m), 1.45 to 1.47 (2H, m), 1.45 to 1.63 (3H, m), 1.85 (2H, m), 3.62 (2H, s), 3.64 (2H, q), 4.05 (1H, d) 4.15 (2H, q), 7.11 (2H, Ar, d), 7.28 to 7.30 (3H, Ar, m), 7.33 (2H, Ar, d), 7.40 (1H, Ar, s).
The activation of receptors with an agonist (activator) in HeLN cells leads to the expression of a reporter gene, luciferase, which, in the presence of a substrate, generates light. The modulation of the receptors is measured as quantity of luminescence produced after incubating the cells in the presence of a reference agonist. The ligands will displace the agonist from its site. The measurement of the activity is performed by quantification of the light produced. This measurement makes it possible to determine the modulatory activity of the compounds according to the invention by determining the constant which is the affinity of the molecule for the receptor. Since this value can fluctuate according to the basal activity and the expression of the receptor, it is called apparent Kd (KdApp in nM).
To determine this constant, “cross curves” for the product to be tested against a reference agonist are produced in a 96-well plate: 10 concentrations of the test product plus a concentration 0 are placed in a line, and 7 concentrations of the agonist plus one concentration 0 are placed in a column. This is 88 measurement points for 1 product and 1 receptor. The 8 remaining wells are used for repeatability controls.
In each well, the cells are in contact with a concentration of the product to be tested and a concentration of the reference agonist, 2-(4-{2-[3-(2,4-difluorophenyl)-1-heptylureido]ethyl}phenylsulfanyl)-2-methylpropionic acid for PPARα, {2-methyl-4-[4-methyl-2-(4-trifluoromethylphenyl)thiazol-5-ylmethylsulfanyl]phenoxy}acetic acid for PPARδ and 5-{4-[2-(methylpyridin-2-ylamino)ethoxy]benzyl}thiazolidine-2,4-dione for PPARγ. Measurements are also carried out for the controls total agonist with the same products.
The HeLN cell lines used are stable transfectants containing the plasmids ERE-βGlob-Luc-SV-Neo (reporter gene) and PPAR (α, δ, γ) Gal-hPPAR. These cells are inoculated into 96-well plates in an amount of 10 000 cells per well in 100 μl of DMEM medium free of phenol red and supplemented with 10% lipid-free calf serum. The plates are then incubated at 37° C., 7% CO2 for 16 hours.
The various dilutions of the test products and of the reference ligand are added in an amount of 5 μl per well. The plates are then incubated for 18 hours at 37° C., 7% CO2. The culture medium is removed by turning over and 100 μl of a 1:1 PBS/Luciferin mixture are added to each well. After 5 minutes, the plates are read by the luminescence reader.
These cross curves make it possible to determine the AC50 values (concentrations at which 50% activation is observed) for the reference ligand at various concentrations of test product. These AC50 values are used to calculate the Schild regression by plotting a straight line corresponding to the Schild equation (“quantitation in receptor pharmacology” Terry P. Kenakin, Receptors and Channels 2001, 7, 371-385) which leads to Kd app values being obtained (in nM).
Transactivation Results:
n.a. means not active
Various specific formulations based on the compounds according to the invention are illustrated in this example.
Each patent, patent application, publication and literature article/report cited or indicated herein is hereby expressly incorporated by reference.
While the invention has been described in terms of various specific and preferred embodiments, the skilled artisan will appreciate that various modifications, substitutions, omissions, and changes may be made without departing from the spirit thereof. Accordingly, it is intended that the scope of the present invention be limited solely by the scope of the following claims, including equivalents thereof.
| Number | Date | Country | Kind |
|---|---|---|---|
| FR 03/50024 | Feb 2003 | FR | national |
This application claims priority under 35 U.S.C. § 119 of FR 03/50024, filed Feb. 12, 2003, and of provisional application Ser. No. 60/452,939, filed Mar. 10, 2003, and is a continuation of PCT/EP 2004/002199, filed Feb. 10, 2004 and designating the United States (published in the English language on Aug. 26, 2004 as WO 2004/072022 A1), each hereby expressly incorporated by reference and each assigned to the assignee hereof.
| Number | Date | Country | |
|---|---|---|---|
| 60452939 | Mar 2003 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/EP04/02199 | Feb 2004 | US |
| Child | 11202059 | Aug 2005 | US |
