Amide derivatives as lysophosphatidic acid receptor antagonists

Abstract

The present invention provides compounds of formula (I) and pharmaceutically acceptable salts thereof, ##STR00001##
wherein R.sup.1, X, m, R.sup.2, Y, R.sup.3, Z, n, R.sup.4, A and B are as defined in the specification, processes for their preparation, pharmaceutical compositions containing them and their use in therapy.

Claims

1. The compound 4-((N-Cyclopropylmethyl)-2-fluoro-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid.

2. The compound 4-((N-Cyclopropylmethyl)-2-fluoro-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition comprising the compound according to claim 2 and a pharmaceutically acceptable adjuvant, diluent or carrier.

4. A pharmaceutical composition comprising the compound according to claim 2 and a pharmaceutically acceptable adjuvant, diluent, or carrier, and at least one additional therapeutic agent.

5. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable adjuvant, diluent or carrier.

6. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable adjuvant, diluent, or carrier, and at least one additional therapeutic agent.

7. A pharmaceutical composition comprising a compound selected from: 4-((N-(cyclopropylmethyl)-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid, 4-((N-(cyclopropylmethyl)-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid, and a pharmaceutically acceptable salt of either of the foregoing; and a pharmaceutically acceptable adjuvant, diluent or carrier.

8. The pharmaceutical composition according to claim 7, wherein the compound is 4-((N-(cyclopropylmethyl)-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid or a pharmaceutically acceptable salt thereof.

9. The pharmaceutical composition according to claim 7, wherein the compound is 4-((N-(cyclopropylmethyl)-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid or a pharmaceutically acceptable salt thereof.

10. A pharmaceutical composition comprising a compound selected from: 4-((N-(cyclopropylmethyl)-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid, 4-((N-(cyclopropylmethyl)-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid, and a pharmaceutically acceptable salt of either of the foregoing; and a pharmaceutically acceptable adjuvant, diluent, or carrier, and at least one additional therapeutic agent.

11. The pharmaceutical composition according to claim 10, wherein the compound is 4-((N-(cyclopropylmethyl)-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid or a pharmaceutically acceptable salt thereof.

12. The pharmaceutical composition according to claim 10, wherein the compound is 4-((N-(cyclopropylmethyl)-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid or a pharmaceutically acceptable salt thereof.

Description

A. PREPARATION OF ACID INTERMEDIATES

(1) ##STR00018##

Intermediate A1: 4-(2-Fluorophenoxy)benzoic acid

(2) ##STR00019##

(3) Step (i) Potassium carbonate (31.1 g, 225 mmol) was added to a solution of 4-fluorobenzaldehyde (7.96 mL, 75 mmol) and 2-fluorophenol (9 mL, 98 mmol) in DMF (100 mL). The mixture was heated at 100 C. under nitrogen for 24 h. After cooling to RT, the mixture was partitioned between EtOAc (200 mL) and water (150 mL). The aqueous phase was extracted with EtOAc (200 mL). The combined organic phases were washed with brine (350 mL), dried and concentrated. The residue was purified by silica chromatography (0-20% EtOAc/petrol) to give 4-(2-fluorophenoxy)benzaldehyde (crude, 17.7 g) which was used in the next step (ii) without further purification or characterisation.

(4) Step (ii) 4-(2-Fluorophenoxy)benzaldehyde (crude, 17.7 g) was dissolved in THF (100 mL), tert-butanol (100 mL) and water (50 mL), and the mixture cooled in an ice-water bath. 2-Methyl-2-butene (43.3 mL, 409 mmol), potassium dihydrogen phosphate (27.8 g, 205 mmol) and sodium chlorite (23.12 g, 205 mmol) were added sequentially and the mixture was stirred at RT for 18 h. The mixture was partitioned between water (100 mL) and Et.sub.2O (200 mL), and the aqueous phase extracted with Et.sub.2O (200 mL). The combined organic phases were washed sequentially with saturated sodium thiosulfate solution (100 mL) and brine (100 mL), dried and concentrated. Addition of EtOAc and petrol resulted in a precipitate which was isolated by filtration and air-dried to give the title compound (11.95 g, 63%) as a pale yellow solid.

(5) MS ES.sup.: 231

Intermediate A2: 4-(2,6-Dimethylphenoxy)benzoic acid

(6) ##STR00020##

(7) Step (i) Tripotassium phosphate (9.27 g, 43.7 mmol) and palladium(II) acetate (0.490 g, 2.18 mmol) were added to a solution of ethyl 4-bromobenzoate (5 g, 21.8 mmol) and 2,6-dimethylphenol (3.73 g, 30.6 mmol) in toluene (40 mL). The mixture was degassed for about 5 minutes and purged with nitrogen. A solution of di-tert-butyl(2,4,6-triisopropylbiphenyl-2-yl)phosphine (1.390 g, 3.27 mmol) in toluene (5 mL) was added. The mixture was degassed for a further 2 minutes, purged with nitrogen, and heated under reflux for 1.5 h. The mixture was partitioned between water and EtOAc (50 mL each), and the organic phase dried and concentrated. The residue was purified by silica chromatography (0-20% EtOAc/petrol) to give ethyl 4-(2,6-dimethylphenoxy)benzoate (crude, 2 g) which was used in the next step (ii) without further purification or characterisation.

(8) Step (ii) Ethyl 4-(2,6-dimethylphenoxy)benzoate (crude, 2 g) was dissolved in MeOH (7 mL). NaOH (2 M, 7 mL) was added, and the mixture heated under microwave irradiation at 120 C. for 20 minutes. The mixture was acidified to pH 1 (2 M HCl), diluted with water (50 mL) and extracted with EtOAc (50 mL). The organic phase was dried and concentrated. The residue was dissolved/suspended in EtOAc (50 mL) and the mother liquor concentrated. This process was repeated with Et.sub.2O:EtOAc (4:1, 20 mL) and then Et.sub.2O (20 mL) to give the title compound (crude, 800 mg) which was used in the next step without further purification or characterisation.

Intermediate A3: 4-(2,6-Difluorophenoxy)benzoic acid

(9) ##STR00021##

(10) Step (i) 4-(2,6-Difluorophenoxy)benzaldehyde was prepared as described for 4-(2-fluorophenoxy)benzaldehyde (Intermediate A1, step (i)) from 4-fluorobenzaldehyde and 2,6-difluorophenol, except with 2 eq potassium carbonate and 1.2 eq of the phenol.

(11) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 7.12-7.21 (m, 2H) 7.32-7.50 (m, 3H) 7.90-7.99 (m, 2H) 9.94 (s, 1H)

(12) Step (ii) 4-(2,6-Difluorophenoxy)benzaldehyde (0.500 g, 2.14 mmol) was dissolved in tert-butanol (6 mL) and water (3 mL). 2-Methyl-2-butene (2 M in THF, 5.34 mL, 10.67 mmol), potassium dihydrogen phosphate (0.73 g, 5.34 mmol) and sodium chlorite (0.72 g, 2.14 mmol) were added sequentially and the mixture was stirred at RT for 4 h. The mixture was partitioned between water (30 mL) and Et.sub.2O (30 mL). The aqueous phase was acidified with HCl (2 M, 10 mL) and extracted with Et.sub.2O (30 mL). The combined organic phases were washed sequentially with saturated sodium thiosulfate solution (15 mL) and brine (15 mL), dried and concentrated. The residue was loaded onto an anion exchange cartridge.

(13) After washing with MeCN, the product was eluted with 2 M HCl/MeCN then concentrated to give the title compound (0.367 g, 69%) as a white solid.

(14) MS ES.sup.: 249

Intermediate A4: 4-(2-Fluoro-6-methylphenoxy)benzoic acid

(15) ##STR00022##

(16) Step (i) 4-(2-Fluoro-6-methylphenoxy)benzaldehyde was prepared as described for 4-(2-fluorophenoxy)benzaldehyde (Intermediate A1, step (i)) from 4-fluorobenzaldehyde and 2-fluoro-6-methylphenol, and used in the next step (ii) without further purification or characterisation.

(17) Step (ii) The title compound was prepared as described for 4-(2-fluorophenoxy)benzoic acid (Intermediate A1, step (ii)) from 4-(2-fluoro-6-methylphenoxy)benzaldehyde (step (i) above), with recrystallisation of the product from EtOAc/petrol, and used in the next step without further purification or characterisation.

Intermediate A5: 4-(2-Chloro-6-fluorophenoxy)benzoic acid

(18) ##STR00023##

(19) Step (i) 4-(2-Chloro-6-fluorophenoxy)benzaldehyde was prepared as described for 4-(2-fluorophenoxy)benzaldehyde (Intermediate A1, step (i)) from 4-fluorobenzaldehyde and 2-chloro-6-fluorophenol.

(20) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 7.07-7.17 (m, 2H) 7.40-7.60 (m, 3H) 7.89-7.99 (m, 2H) 9.93 (s, 1H)

(21) Step (ii) The title compound was prepared as described for 4-(2-fluorophenoxy)benzoic acid (Intermediate A1, step (ii)) from 4-(2-chloro-6-fluorophenoxy)benzaldehyde (step (i) above).

(22) MS ES.sup.: 265

Intermediate A6: 2-Fluoro-4-(2-fluorophenoxy)benzoic acid

(23) ##STR00024##

(24) Step (i) 2-Fluoro-4-(2-fluorophenoxy)benzaldehyde was prepared as described for 4-(2-fluorophenoxy)benzaldehyde (Intermediate A1, step (i)) from 2,4-difluorobenzaldehyde and 2-fluorophenol, except with 1 eq of the phenol and heating at 80 C.

(25) MS ES.sup.+: 235

(26) Step (ii) The title compound was prepared as described for 4-(2-fluorophenoxy)benzoic acid (Intermediate A1, step (ii)) from 2-fluoro-4-(2-fluorophenoxy)benzaldehyde (step (i) above).

(27) MS ES.sup.+: 251

Intermediate A7: 4-((2-Fluorobenzyl)oxy)benzoic acid

(28) ##STR00025##

(29) Step (i) Sodium hydride (60% dispersion in mineral oil, 0.177 g, 4.43 mmol) was added to a mixture of 4-fluorobenzaldehyde (0.437 mL, 4.03 mmol) and (2-fluorophenyl)methanol (0.508 g, 4.03 mmol) in DMF (20 mL). The reaction was stirred under nitrogen at room temperature for 18 h. The mixture was diluted with EtOAc and washed with sodium bicarbonate and then brine. The organic phase was dried and concentrated to give 4-((2-fluorobenzyl)oxy)benzaldehyde as a residue which was used in the next step (ii) without further purification or characterisation.

(30) Step (ii) The title compound was prepared as described for 4-(2,6-difluorophenoxy)benzoic acid (Intermediate A3, step (ii)) from 4-((2-fluorobenzyl)oxy)benzaldehyde (step (i) above) and used in the next step without further purification or characterisation.

Intermediate A8: 2-Fluoro-4-(2-methoxyphenoxy)benzoic acid

(31) ##STR00026##

(32) Prepared by a two step process analogous to that described for Intermediate A1 in which, in the first step, 2,4-difluorobenzaldehyde was reacted with 2-methoxyphenol to yield 2-fluoro-4-(2-methoxyphenoxy)benzaldehyde which, without further purification or characterisation, was used in the second oxidation step to yield the titled compound.

(33) The intermediates 4-(2-methylphenoxy)benzoic acid and 4-(2-chlorophenoxy)benzoic acid used in the preparation of several of the final compounds is known in the art and is commercially available from chemical suppliers such as Apollo Scientific.

B. PREPARATION OF 4-(AMINOMETHYL)BENZOATES AND ANALOGUES

(34) ##STR00027##

(35) Intermediates of formula (B) shown in Table 1 below were prepared according to one of the following methods R1 to R6:

(36) Method R1

(37) A solution of the amine (50 mmol) and methyl 4-formylbenzoate (50 mmol) in IMS or EtOH (100 mL) was heated under reflux for 2 h. After cooling, the mixture was concentrated and the residue was dissolved in MeOH (400 mL). The resulting solution was cooled in an ice-water bath and sodium borohydride (40 mmol) was added, typically in several portions over 10 minutes. The mixture was stirred and allowed to warm to RT. After 2 h, the mixture was concentrated. The residue was stirred in water (20 mL). The mixture was acidified HCl (2 M) and extracted with DCM (320 mL). The aqueous phase was basified with solid NaOH and extracted with DCM (350 mL). The latter extracts were dried and concentrated to give the intermediate of formula (B).

(38) Method R2

(39) As for Method R1, except that after concentrating the sodium borohydride reaction, the residue was dissolved/suspended in HCl (2 M) or HCl (4 M in 1,4-dioxane). The resulting precipitate was filtered, washed with water, and dried to give the hydrochloride salt of the intermediate of formula (B).

(40) Method R3

(41) A solution of the amine (5 mmol) and methyl 4-formylbenzoate (5 mmol) was stirred in DCM (20 mL) for 2 h at RT. Sodium triacetoxyborohydride (10 mmol) was added, followed by acetic acid (6 mmol). After stirring for 2 h, the mixture was diluted with DCM (20 mL) and washed with NaOH (2 M, 20 mL) or saturated sodium bicarbonate solution (20 mL). The aqueous phase was extracted with DCM (20 mL), and the combined organic phases dried and concentrated to give the intermediate of formula (B).

(42) Method R4

(43) As for Method R1, except that after concentrating the sodium borohydride reaction and adding water and HCl, the mixture was basified with sodium hydrogencarbonate and extracted with EtOAc. The organic phase was dried and concentrated, and the residue was dissolved/suspended in an organic solvent, typically EtOAc or Et.sub.2O. HCl (4 M in MeOH) was added, and the mixture stirred at RT. The resulting precipitate was filtered, and the filter cake air-dried to give the hydrochloride salt of the intermediate of formula (B).

(44) Method R5

(45) As for Method R1, except that (a) in the imine formation, triethylamine (typically 1 eq) was added, and (b) after concentrating the sodium borohydride reaction, the residue was partitioned between (i) sodium hydrogencarbonate or sodium hydroxide and (ii) EtOAc or DCM. The organic phase was dried and concentrated to give the intermediate of formula (B). In some cases, the residue was dissolved/suspended in an organic solvent, typically EtOAc or Et.sub.2O. HCl (4 M in MeOH) was added, and the mixture stirred at RT. The resulting precipitate was filtered, and the filter cake air-dried to give the hydrochloride salt of the intermediate of formula (B). In some cases, the residue was dissolved/suspended in aqueous HCl (2 M). The resulting precipitate was filtered, washed with water, and dried to give the hydrochloride salt of the intermediate of formula (B).

(46) Method R6

(47) As for Method R1, except that after stirring the sodium borohydride reaction for 1 h, the mixture was quenched with HCl (2 M). The resulting precipitate was isolated by filtration and dried under vacuum to give the hydrochloride salt of the intermediate of formula (B).

(48) TABLE-US-00001 TABLE 1 (B) Inter- mediate R Product Method Data B1 CH.sub.2CH.sub.3 methyl 4- R1, using MS ((ethylamino)methyl)benzoate 2 M ES.sup.+: EtNH.sub.2 in 194 THF (1.8 eq) B2 CH.sub.2CHF.sub.2 methyl 4-(((2,2- R1 MS difluoroethyl)amino)methyl) ES.sup.+: benzoate 230 B3 CH.sub.2CF.sub.3 methyl 4-(((2,2,2- R2, using MS (HCl trifluoroethyl)amino)methyl) 5 eq ES.sup.+: salt) benzoate hydrochloride NaBH.sub.4 248 B4 CH.sub.2iPr methyl 4- R4 n/a (HCl ((isobutylamino)methyl) salt) benzoate hydrochloride B5 (HCl salt) methyl 4- (((cyclopropylmethyl)amino) methyl)benzoate hydrochloride R4 MS ES.sup.+: 220 B6 (HCl salt) 0 methyl 4- (((cyclobutylmethyl)amino) methyl)benzoate hydrochloride R5, from Amine 1* MS ES.sup.+: 234 B7 (HCl salt) methyl 4- ((isopentylamino)methyl) benzoate hydrochloride R4 MS ES.sup.+: 236 B8 (HCl salt) methyl 4-(((2- cyclopropylethyl)amino) methyl)benzoate hydrochloride R4, from Amine 2** MS ES.sup.+: 234 B9 (HCl salt) methyl 4-(((2- cyclobutylethyl)amino)methyl) benzoate hydrochloride R4, from Amine 3*** MS ES.sup.+: 248 B10 (HCl salt) methyl 4- ((benzylamino)methyl) benzoate hydrochloride R4 MS ES.sup.+: 256 B11 (HCl salt) methyl 4-(((2- fluorobenzyl)amino)methyl) benzoate hydrochloride R2 MS ES.sup.+: 274 B12 (HCl salt) methyl 4-(((3- methoxybenzyl)amino)methyl) benzoate hydrochloride R2 MS ES.sup.+: 286 B13 (HCl salt) methyl 4-(((4- methoxybenzyl)amino)methyl) benzoate hydrochloride R2 MS ES.sup.+: 286 B14 methyl 4- ((phenethylamino)methyl) benzoate R3 MS ES.sup.+: 270 B15 (HCl salt) methyl 4-(((3- phenylpropyl)amino)methyl) benzoate hydrochloride R6 MS ES.sup.+: 284 B16 0 methyl 4-((((trans-2- phenylcyclopropyl)methyl) amino)methyl)benzoate R3 MS ES.sup.+: 296 B17 (HCl salt) methyl 4-(((3-(2- fluorophenyl)propyl)amino) methyl)benzoate hydrochloride R5 MS ES.sup.+: 302 B18 (HCl salt) methyl 4-(((3-(3- fluorophenyl)propyl)amino) methyl)benzoate hydrochloride R5 MS ES.sup.+: 302 B19 (HCl salt) methyl 4-(((4- phenylbutyl)amino)methyl) benzoate hydrochloride R4 n/a B19a CH.sub.2CH.sub.2CH.sub.3 methyl 4- R2 n/a (HCl ((propylamino)methyl) salt) benzoate hydrochloride B19b (HCl salt) methyl 4-(((2,2- difluoropropyl)amino)methyl) benzoate R5 MS ES.sup.+: 244 B19c methyl 4-(((3,3,3- trifluoropropyl)amino)methyl R3 MS ES.sup.+: benzoate 262 B19d (HCl methyl 4- ((butylamino)methyl)benzoate R2 n/a salt) hydrochloride

*Amine 1: Cyclobutylmethanamine Hydrochloride

(49) A solution of borane in THF (1 M, 296 mL, 296 mmol) was added to a solution of cyclobutanecarbonitrile (20 g, 247 mmol) in THF (60 mL) under argon, and the mixture heated under reflux overnight. The mixture was cooled in an ice-water bath and MeOH (120 mL) added dropwise while keeping the temperature of the mixture below 20 C. HCl in MeOH (4 M, 300 mL) was added, again keeping the temperature below 20 C. The resulting solution was heated under reflux for 2.5 h. After cooling, the mixture was concentrated. The residue was diluted with MeOH (100 mL) and concentrated, and this process was repeated. Ether was added to the residue, and the mixture stirred for 30 minutes before filtering to give the title compound (25.9 g, 86%) as a white solid.

(50) .sup.1H NMR (300 MHz, DMSO-d.sub.6) ppm 1.70-1.86 (m, 4H) 1.97-2.04 (m, 2H) 2.46-2.61 (m, 1H) 2.75-2.84 (m, 2H) 8.02 (br s, 3H)

**Amine 2: 2-Cyclopropylethanamine

(51) Sulfuric acid (17.25 mL, 324 mmol) was added dropwise to a suspension of lithium aluminium hydride (24.56 g, 647 mmol) in Et.sub.2O (900 mL) in an ice-water bath under argon. The mixture was stirred at RT for 1 h. A solution of 2-cyclopropylacetonitrile (17.5 g, 216 mmol) in Et.sub.2O (100 mL) was added and the mixture heated under reflux for 18 h. The mixture was cooled in an ice-water bath and sodium sulfate decahydrate was added until effervescence ceased. The mixture was filtered and concentrated to give the title compound (crude, 18.6 g) which was used without further purification or characterisation.

***Amine 3: 2-Cyclobutylethanamine

(52) Prepared as described for Amine 2 from 2-cyclobutylacetonitrile and used in the next step without further purification or characterisation.

Intermediate B20: (S)-methyl 4-(((2-hydroxy-3-phenylpropyl)amino)methyl)benzoate

(53) ##STR00047##

(54) Step (i) (S,S)-(+)-N,N-Bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediaminocobalt(II) (0.048 g, 0.080 mmol), THF (0.25 mL) and acetic acid (0.018 mL, 0.318 mmol) were added to 2-benzyloxirane (4.27 g, 31.8 mmol). The mixture was cooled in an ice-water bath and water (0.129 mL, 7.16 mmol) was added in one portion. After stirring at RT overnight, the mixture was distilled under vacuum (2 mmHg, bp 60-80 C.) to give (S)-2-benzyloxirane (crude, 2.74 g) which was used in the next step without further purification or characterisation.

(55) Step (ii) (S)-2-Benzyloxirane (step (i) above) (2.74 g, 20.42 mmol) was added to a solution of methyl 4-(aminomethyl)benzoate hydrochloride (7.41 g, 36.8 mmol) and DIPEA (8.92 mL, 51.1 mmol) in MeOH (300 mL). After stirring for 3 days, the reaction mixture was concentrated. The residue was partitioned between saturated sodium hydrogencarbonate solution (200 mL) and DCM (2200 mL). The aqueous phase was extracted with DCM (200 mL). The combined organic phases were concentrated and purified by silica chromatography (0-10% MeOH/DCM) and then reverse-phase chromatography on C18 silica (eluted with 0-95% MeOH/H.sub.2O with 0.1% NH.sub.4OH) to give the title compound (3.27 g, 53%) as a pale brown gum.

(56) MS ES.sup.+: 300

Intermediate B21: (R)-methyl 4-(((2-hydroxy-3-phenylpropyl)amino)methyl)benzoate

(57) ##STR00048##

(58) Step (i) Prepared as described for (S)-2-benzyloxirane (Intermediate B48, step (i)), except using (R,R)-()-N,N-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediaminocobalt(II) to give (R)-2-benzyloxirane which was used in the next step without further purification or characterisation.

(59) Step (ii) Prepared as described for (S)-methyl 4-(((2-hydroxy-3-phenylpropyl)amino)methyl)benzoate (Intermediate B20, step (ii)) from (R)-2-benzyloxirane (step (i) above) and methyl 4-(aminomethyl)benzoate hydrochloride to give the title compound as a pale brown gum.

(60) MS ES.sup.+: 300

C. PREPARATION OF INTERMEDIATE ESTERS

(61) Intermediates of formula (C) shown in Tables 2 to 4 below were prepared according to one of the following methods E1 to E5:

(62) ##STR00049##

(63) Method E1

(64) Triethylamine (6 mmol), 1-hydroxy-7-azabenzotriazole (2.4 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (2.4 mmol) were added to a solution/suspension of the relevant amine or amine hydrochloride (2 mmol) and 4-phenoxybenzoic acid (2 mmol) in THF (10 mL). The reaction was stirred at RT for 18 h. (In cases where the amine hydrochloride was used, an extra 2 mmol triethylamine was added.) The mixture was partitioned between (i) water or saturated sodium hydrogencarbonate solution (10 mL) and (ii) DCM or EtOAc (10 mL). The aqueous phase was extracted with further DCM or EtOAc, and the combined organic phases dried and concentrated. The residue was purified by silica chromatography (typically 0-100% EtOAc/petrol).

(65) Method E2

(66) DIPEA (2.2 mmol) and HBTU (1.1 mmol) were added to a solution of the relevant acid in DMF (5 mL) and the mixture stirred for 5 minutes at RT. The relevant amine or amine hydrochloride (0.95 mmol) and DBU (1.5 mmol) were added. The mixture was stirred for 1 h at RT. (In cases where the amine hydrochloride was used, an extra 1 mmol DIPEA was added.) The mixture was partitioned between EtOAc (30 mL) and either water or saturated sodium hydrogencarbonate solution (30 mL). In some cases the aqueous phase was further extracted with EtOAc. The combined organic phases were dried and concentrated. The residue was purified by silica chromatography (typically 0-100% EtOAc/petrol).

(67) Method E3

(68) Thionyl chloride or oxalyl chloride (3-12 mmol) was added to a solution of the acid (3 mmol) in DCM (10 mL) in an ice-water bath. In some cases, several drops of DMF were added. The reaction mixture was stirred at RT for 4 h and then concentrated to yield the crude acid chloride, which was dissolved in THF (3 mL) and added to a solution/suspension of the amine hydrochloride (2 mmol) and potassium carbonate (10 mmol) in THF (10 mL). The reaction was heated under reflux for 18 h. After cooling, the mixture was partitioned between water and EtOAc. The organic phase was dried and concentrated and the residue purified by silica chromatography (0-50% EtOAc/petrol).

(69) Method E4

(70) HATU (1.1 mmol) was added to a solution of the acid (1 mmol) and DIPEA (2.2 mmol) in DMF or THF (2 mL). The mixture was stirred at RT and allowed to stand. After 5 minutes, the relevant amine or amine hydrochloride (1.1 mmol) was added. (In some cases, 0.9 mmol of amine or amine hydrochloride was used.) When the amine hydrochloride was used, DBU (1.5 mmol) was added at this stage. After 10 minutes, the mixture was diluted with EtOAc or DCM (30 mL) and washed with either HCl (1 M, 20 mL) or saturated sodium bicarbonate solution (20 mL) or water (20 mL). In some cases, the aqueous phase was further extracted with EtOAc or DCM. The combined organic phases were washed with water (220 mL), dried and concentrated. In some cases further purification was carried out, by silica chromatography (typically 0-100% EtOAc/petrol), reverse-phase chromatography on C18 silica (typically eluting with 5-95% MeCN/water) or preparative HPLC.

(71) ##STR00050##

(72) Method E5

(73) Step (i) Triethylamine (1.351 mL, 9.69 mmol) was added slowly to a mixture of 4-bromobenzoyl chloride (0.851 g, 3.88 mmol) and methyl 4-((phenethylamino)methyl)-benzoate (Intermediate B14) (0.870 g, 3.23 mmol) in DCM (20 mL) at RT under nitrogen and the mixture stirred for 18 h. The mixture was diluted with DCM (30 mL), water (10 mL) and HCl (2 M, 10 mL), and the phases separated. The aqueous phase was extracted with DCM (30 mL) and the combined organic phases washed with water (10 mL), dried and concentrated. The residue was purified by silica chromatography (0-50% EtOAc/petrol) to give methyl 4-((4-bromo-N-phenethylbenzamido)methyl)benzoate (1.21 g, 83%) as a pale orange oil.

(74) MS ES.sup.+: 452, 454

(75) Step (ii) Tripotassium phosphate (2 mmol), palladium(II) acetate (0.1 mmol) and di-tert-butyl(2,4,6-triisopropylbiphenyl-2-yl)phosphine (0.15 mmol) were added to a solution of methyl 4-((4-bromo-N-phenethylbenzamido)methyl)benzoate (step (i) above) (1 mmol) and the relevant phenol (1.2 mmol) in toluene (5 mL). The mixture was degassed for about 5 minutes, purged with nitrogen and heated either thermally or under microwave irradiation at 120 C. for 20 h. The mixture was filtered through diatomaceous earth, washing through with EtOAc. The filtrate was concentrated and purified either by silica chromatography (typically 0-100% EtOAc/petrol) or preparative HPLC.

(76) TABLE-US-00002 TABLE 2 Inter- mediate R Product Method Data C6 CH.sub.2CH.sub.3 methyl 4-((N-ethyl-4-(2- E2 MS ES.sup.+: fluorophenoxy)benzamido) 408 methyl)benzoate C7 CH.sub.2CHF.sub.2 methyl 4-((N-(2,2- E2 MS ES.sup.+: difluoroethyl)-4-(2- 444 fluorophenoxy)benzamido) methyl)benzoate C8 CH.sub.2CF.sub.3 methyl 4-((4-(2- E3 MS ES.sup.+: fluorophenoxy)-N-(2,2,2- 462 trifluoroethyl)benzamido) methyl)benzoate C9 CH.sub.2iPr methyl 4-((4-(2- E1 MS ES.sup.+: fluorophenoxy)-N- 436 isobutylbenzamido)methyl) benzoate C10 methyl 4-((N- (cyclopropylmethyl)-4-(2- fluorophenoxy)benzamido) methyl)benzoate E1 MS ES.sup.+: 434 C11 methyl 4-((N- (cyclobutylmethyl)-4-(2- fluorophenoxy)benzamido) methyl)benzoate E4 MS ES.sup.+: 448 C12 methyl 4-((4-(2- fluorophenoxy)-N- isopentylbenzamido)methyl) benzoate E1 MS ES.sup.+: 450 C13 methyl 4-((N-(2- cyclopropylethyl)-4-(2- fluorophenoxy)benzamido) methyl)benzoate E1 MS ES.sup.+: 448 C14 methyl 4-((N-(2- cyclobutylethyl)-4-(2- fluorophenoxy)benzamido) methyl)benzoate E1 MS ES.sup.+: 462 C15 methyl 4-((N-benzyl-4-(2- fluorophenoxy)benzamido) methyl)benzoate E1 MS ES.sup.+: 470 C16 methyl 4-((N-(2- fluorobenzyl)-4-(2- fluorophenoxy)benzamido) methyl)benzoate E4 MS ES.sup.+: 488 C17 methyl 4-((4-(2- fluorophenoxy)-N-(3- methoxybenzyl)benzamido) methyl)benzoate E4 n/a C18 0 methyl 4-((4-(2- fluorophenoxy)-N-(4- methoxybenzyl)benzamido) methyl)benzoate E4 n/a C19 methyl 4-((4-(2- fluorophenoxy)-N- phenethylbenzamido)methyl) benzoate E4 MS ES.sup.+: 484 C20 methyl 4-((4-(2- fluorophenoxy)-N-(3-(2- fluorophenyl)propyl) benzamido)methyl)benzoate E4 MS ES.sup.+: 516 C21 methyl 4-((4-(2- fluorophenoxy)-N-(3-(3- fluorophenyl)propyl) benzamido)methyl)benzoate E4 MS ES.sup.+: 516 C22 methyl 4-((4-(2- fluorophenoxy)-N-((trans-2- phenylcyclopropyl)methyl) benzamido)methyl)benzoate E4 MS ES.sup.+: 510 C23 (S)-methyl 4-((4-(2- fluorophenxoy)-N-(2- hydroxy-3- phenylpropyl)benzamido) methyl)benzoate E4 MS ES.sup.+: 514 C24 (R)-methyl 4-((4-(2- fluorophenoxy)-N-(2- hydroxy-3- phenylpropyl)benzamido) methyl)benzoate E4 MS ES.sup.+: 514 C25 methyl 4-((4-(2- fluorophenoxy)-N-(4- phenylbutyl)benzamido) methyl)benzoate E2 MS ES.sup.+: 512 C25a CH.sub.2CH.sub.2CH.sub.3 methyl 4-((4-(2- E4 MS ES.sup.+: fluorophenoxy)-N- 422 propylbenzamido)- methyl)benzoate C25b CH.sub.2CH.sub.2CF.sub.3 methyl 4-((4-(2- E4 MS ES+: fluorophenoxy)-N-(3,3,3- n/a trifluoropropyl)benzamido) methyl)benzoate C25c CH.sub.2CF.sub.2CH.sub.3 methyl 4-((N-(2,2- E3 MS ES+: difluoropropyl)-4-(2- n/a fluorophenoxy)benzamido) methyl)benzoate C25d methyl 4-((N-butyl-4-(2- fluorophenoxy)benzamido) methyl)benzoate E4 MS ES+: 436

(77) TABLE-US-00003 TABLE 3 Inter- mediate X R Product Method Data C26 Me 0 methyl 4-((N- (cyclopropylmethyl)-4-(o- tolyloxy)benzamido)methyl) benzoate E4 MS ES.sup.+: 430 C27 Me methyl 4-((N- (cyclobutylmethyl)-4-(o- tolyloxy)benzamido)methyl) benzoate E4 MS ES.sup.+: 444 C28 Me methyl 4-((N-benzyl-4-(o- tolyloxy)benzamido)methyl) benzoate E4 MS ES.sup.+: 466 C29 Me methyl 4-((N-phenethyl-4-(o- tolyloxy)benzamido)methyl) benzoate E5 MS ES.sup.+: 480 C30 CF.sub.3 methyl 4-((N-phenethyl-4-(2- (trifluoromethyl)phenoxy) benzamido)-methyl)benzoate E5 MS ES.sup.+: 534 C31 CN methyl 4-((N-benzyl-4-(-2 cyanophenoxy)benzamido) methyl)-benzoate E4 MS ES.sup.+: 477 C32 OMe CH.sub.2CH.sub.3 methyl 4-((N-ethyl-4-(2- E2 MS ES.sup.+: methoxyphenoxy)benzamido) 420 methyl)-benzoate C33 OMe CH.sub.2CHF.sub.2 methyl 4-((N-(2,2- E2 MS ES.sup.+: difluoroethyl)-4-(2- 456 methoxyphenoxy)benzamido) methyl)-benzoate C34 OMe CH.sub.2iPr methyl 4-((N-isobutyl-4-(2- E4 MS ES.sup.+: methoxyphenoxy)benzamido) 448 methyl)-benzoate C35 OMe methyl 4-((N- (cyclopropylmethyl)-4-(2- methoxyphenoxy)benzamido) methyl)-benzoate E4 MS ES.sup.+: 446 C36 OMe methyl 4-((4-(2- methoxyphenoxy)-N-(3- phenylpropyl)benzamido)- methyl)benzoate E1 MS ES.sup.+: 510 C37 OMe methyl 4-((N-(3-(3- fluorophenyl)propyl)-4-(2- methoxyphenoxy)benzamido) methyl)-benzoate E1 MS ES.sup.+: 528 C38 OEt methyl 4-((N- (cyclopropylmethyl)-4-(2- ethoxyphenoxy)benzamido) methyl)-benzoate E5 MS ES.sup.+: 460 C39 Cl 0 methyl 4-((4-(2- chlorophenoxy)-N- (cyclopropylmethyl) benzamido)methyl)benzoate E4 MS ES.sup.+: 450 C40 Cl methyl 4-((4-(2- chlorophenoxy)-N- phenethylbenzamido)methyl) benzoate E5 MS ES.sup.+: 500 C40a OMe CH.sub.2CF.sub.3 methyl 4-((4-(2- E3 MS ES.sup.+: methoxyphenoxy)-N-(2,2,2- n/a trifluoroethyl)benzamido)- methyl)benzoate C40b OMe CH.sub.2CH.sub.2CH.sub.3 methyl 4-((4-(2- E4 MS ES.sup.+: methoxyphenoxy)-N- n/a propylbenzamido)methyl) benzoate C40c OMe CH.sub.2CF.sub.2CH.sub.3 methyl 4-((N-(2,2- E3 MS ES.sup.+: difluoropropyl)-4-(2- n/a methoxyphenoxy)benzamido) methyl)-benzoate C40d OMe CH.sub.2CH.sub.2CF.sub.3 methyl 4-((N-(3,3,3- E4 MS ES.sup.+: trifluoropropyl)-4-(2- 488 methoxyphenoxy)benzamido)- methyl)benzoate C40e OMe methyl 4-((N-butyl-4-(2- methoxyphenoxy)benzamido) E4 MS ES.sup.+: n/a methyl)-benzoate C40f Me CH.sub.2CF.sub.3 methyl 4-((4-(o-tolyloxy)-N- E3 MS ES+: (2,2,2- n/a trifluoroethyl)benzamido) methyl)-benzoate C40g Cl CH.sub.2CF.sub.3 methyl 4-((4-(2- E3 MS ES+: chlorophenoxy)-N-(2,2,2- n/a trifluoroethyl)benzamido)- methyl)benzoate C40h Me CH.sub.2CH.sub.3 methyl 4-((N-ethyl-4-(o- E4 MS ES+: tolyloxy)benzamido)methyl) n/a benzoate C40i Cl CH.sub.2CH.sub.3 methyl 4-((4-(2- E4 MS ES+: chlorophenoxy)-N- n/a ethylbenzamido)methyl) benzoate C40j Cl CH.sub.2CHF.sub.2 methyl 4-((4-(2- E3 MS ES.sup.+: chlorophenoxy)-N-(2,2- n/a difluoroethyl)benzamido) methyl)-benzoate

(78) TABLE-US-00004 TABLE 4 Inter- mediate R.sup.1 R Product Method Data C41 methyl 4-((N- (cyclopropylmethyl)- 4-(2,6- difluorophenoxy)- benzamido)methyl) benzoate E1 MS ES.sup.+: 452 C42 methyl 4-((N-(2- cyclopropylethyl)-4- (2,6- difluorophenoxy)- benzamido)methyl) benzoate E1 MS ES.sup.+: 466 C43 methyl 4-((N-benzyl- 4-(2,6- difluorophenoxy) benzamido)- methyl)benzoate E1 MS ES.sup.+: 488 C44 0 methyl 4-((N- (cyclopropylmethyl)- 4-(2-fluoro-6- methylphenoxy) benzamido)- methyl)benzoate E4 MS ES.sup.+: 448 C45 methyl 4-((N- (cyclobutylmethyl)- 4-(2-fluoro-6- methylphenoxy)- benzamido)methyl) benzoate E4 MS ES.sup.+: 462 C46 methyl 4-((4-(2- chloro-6- fluorophenoxy)-N- (cylcopropylmethyl) benzamido)- methyl)benzoate E1 MS ES.sup.+: 468 C47 methyl 4-((4-(2- chloro-6- fluorophenoxy)-N- (2-cyclopropylethyl) benzamido)- methyl)benzoate E1 MS ES.sup.+: 482 C48 methyl 4-((N-benzyl- 4-(2-chloro-6- fluorophenoxy) benzamido)methyl)- benzoate E1 MS ES.sup.+: 504 C49 00 01 methyl 4-((4-(2,6- dimethylphenoxy)- N- isopentylbenzamido) methyl)-benzoate E4 MS ES.sup.+: 460 C50 02 03 methyl 4-((N-benzyl- 4-(2,6- dimethylphenoxy) benzamido)- methyl)benzoate E4 MS ES.sup.+: 480 C51 04 05 methyl 4-((4-(2,6- dimethylphenoxy)- N-phenethyl benzamido)methyl)- benzoate E4 MS ES.sup.+: 494 C52 06 07 methyl 4-((4-(3- methoxyphenoxy)- N-phenethyl benzamido)methyl)- benzoate E5 MS ES.sup.+: 496 C53 08 09 methyl 4-((4-(4- methoxyphenoxy)- N-phenethyl benzamido)- methyl)benzoate E5 MS ES.sup.+: 496 C54 0 methyl 4-((4-(3- chlorophenoxy)-N- phenethyl benzamido)methyl)- benzoate E5 MS ES.sup.+: 500 C55 methyl 4-((N- (cyclopropylmethyl)- 4-((2- fluorobenzyl)oxy) benzamido)- methyl)benzoate E4 MS ES.sup.+: 448 C56 methyl 4-((N- (cyclobutylmethyl)- 4-((2- fluorobenzyl)oxy) benzamido)- methyl)benzoate E4 MS ES.sup.+: 462 C57 methyl 4-((N-benzyl- 4-((2- fluorobenzyl)oxy) benzamido)- methyl)benzoate E4 MS ES.sup.+: 484

Intermediate C58: Methyl 4-((4-(2-fluorophenoxy)-N-((3-methoxycyclobutyl)methyl)benzamido)methyl)benzoate (mixture of cis and trans ring isomers)

(79) ##STR00118##

(80) Step (i) Prepared using the HATU coupling procedure described in Method E4 above from 3-methoxycyclobutanecarboxylic acid (500 mg, 3.84 mmol) and (4-bromophenyl)methanamine hydrochloride (940 mg, 4.23 mmol) to give N-(4-bromobenzyl)-3-methoxycyclobutanecarboxamide (mixture of cis and trans ring isomers) (880 mg, 77%).

(81) MS ES.sup.+: 298, 300

(82) Step (ii) Lithium aluminium hydride (1 M solution in THF, 2.62 mL, 2.62 mmol) was added to a solution of N-(4-bromobenzyl)-3-methoxycyclobutanecarboxamide from step (i) above (780 mg, 2.62 mmol) in THF (20 mL) at RT. After stirring for 18 h, the mixture was quenched with saturated sodium sulfate dodecahydrate solution (5 mL), filtered and concentrated. The residue was dissolved in DCM (50 mL) and washed with NaOH (2 M, 50 mL). The organic phase was concentrated and the residue purified by silica chromatography (0-30% EtOAc/petrol) to give N-(4-bromobenzyl)-1-(3-methoxycyclobutyl)methanamine (mixture of cis and trans ring isomers) which was used in the next step without further purification or characterisation.

(83) Step (iii) Prepared using the HATU coupling procedure described in Method E4 above from 4-(2-fluorophenoxy)benzoic acid (Intermediate A1) (310 mg, 1.337 mmol) and N-(4-bromobenzyl)-1-(3-methoxycyclobutyl)methanamine from step (ii) above (380 mg, 1.337 mmol) to give the desired compound, N-(4-bromobenzyl)-4-(2-fluorophenoxy)-N-((3-methoxycyclobutyl)methyl)benzamide (mixture of cis and trans ring isomers), along with the corresponding des-bromo analogue N-benzyl-4-(2-fluorophenoxy)-N-((3-methoxycyclobutyl)methyl)benzamide (also a mixture of cis and trans ring isomers). This material (approximately 1:1 mixture of bromo:des-bromo compound, 684 mg in total) was used in the next step without further purification. MS ES.sup.+: 498, 500

(84) Step (iv) A solution of palladium(II) acetate (27.0 mg, 0.120 mmol), 1,3-bis(diphenylphosphino)propane (49.7 mg, 0.120 mmol), and N-(4-bromobenzyl)-4-(2-fluorophenoxy)-N-((3-methoxycyclobutyl)methyl)benzamide from step (iii) above (300 mg, 0.602 mmol) in MeOH (5 mL) and DMF (3 mL) was degassed and purged with nitrogen. Triethylamine (0.168 mL, 1.204 mmol) was added, the reaction mixture was evacuated and carbon monoxide passed through the solution. After stirring under carbon monoxide at 70 C. for 18 h, the mixture was MeOH-evaporated and the residue dissolved in DCM (20 mL). The organic phase was washed with water (20 mL) and concentrated. The resulting residue was purified by silica chromatography (0-30% EtOAc/petrol) to give the title compound (115 mg, 80% based on starting material being 50% pure).

(85) MS ES.sup.+: 478

Intermediate C59: Methyl 4-((N-(cyclopropylmethyl)-2-fluoro-4-(2-fluorophenoxy)benzamido)methyl)benzoate

(86) ##STR00119##

(87) Prepared using the EDC coupling procedure described in Method E1 above from 2-fluoro-4-(2-fluorophenoxy)benzoic acid (Intermediate A6) and methyl 4-((cyclopropylmethylamino)methyl)benzoate hydrochloride (Intermediate B5).

(88) MS ES.sup.+: 452

Intermediate C60: Methyl 4-((N-benzyl-4-(2-fluorophenoxy)-cyclohexanecarboxamido)methyl)benzoate (ca. 1:1 mixture of cis and trans ring isomers)

(89) ##STR00120##

(90) Step (i) 4-Toluenesulfonyl chloride (18.29 g, 96 mmol) was added in several portions to a solution of ethyl 4-hydroxycyclohexanecarboxylate (ca. 1:1 mixture of cis and trans ring isomers) (29.5 g, 90 mmol) in pyridine (100 mL) in an ice-water bath, and the mixture stirred, allowing to warm to RT. After the solid had dissolved, the mixture was allowed to stand. After 24 h, the mixture was concentrated and the residue partitioned between water and EtOAc (100 mL each). The organic phase was dried and concentrated to give ethyl 4-(tosyloxy)cyclohexanecarboxylate (ca. 1:1 mixture of cis and trans ring isomers) (crude, 29.5 g) as a colourless oil which was used in the next step without further purification.

(91) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.20-1.30 (m, 3H) 1.44-2.08 (m, 8H) 2.21-2.43 (m, 1H) 2.48 (s, 3H) 4.08-4.20 (m, 2H) 4.40-4.50 and 4.70-4.78 (both m, total 1H) 7.28-7.40 (m, 2H) 7.78-7.86 (m, 2H)

(92) Step (ii) 2-Fluorophenol (2.145 mL, 23.25 mmol) was added to a solution/suspension of ethyl 4-(tosyloxy)cyclohexanecarboxylate from step (i) above and cesium carbonate (7.58 g, 23.25 mmol) in DMF (100 mL) and the mixture stirred for 18 h at 80 C. After cooling, the mixture was diluted with EtOAc (200 mL) and washed with saturated sodium hydrogencarbonate solution (100 mL) and brine (100 mL). The organic phase was dried and concentrated, and the residue purified by silica chromatography (0-100% EtOAc/petrol) to give ethyl 4-(2-fluorophenoxy)cyclohexanecarboxylate (ca. 1:1 mixture of cis and trans ring isomers) (crude, 3.41 g) which was used without further purification or characterisation.

(93) Step (iii) Lithium hydroxide (0.307 g, 12.80 mmol) was added to a solution of ethyl 4-(2-fluorophenoxy)cyclohexanecarboxylate from step (ii) above (crude, 3.41 g) in 1,4-dioxane (30 mL) and water (30 mL), and the mixture heated under microwave irradiation at 100 C. for 15 minutes. The mixture was concentrated. The residue was loaded onto an anion exchange cartridge. After washing with MeCN, the product was eluted with 2 M HCl/MeCN and concentrated to give 4-(2-fluorophenoxy)cyclohexanecarboxylic acid (ca. 1:1 mixture of cis and trans ring isomers) (crude, 1.0 g) as a pale yellow solid which was used in the next step without further purification.

(94) MS ES.sup.: 237

(95) Step (iv) The title compound was prepared using the HATU coupling procedure described in Method E4 above, from 4-(2-fluorophenoxy)cyclohexanecarboxylic acid from step (iii) above and methyl 4-((benzylamino)methyl)benzoate hydrochloride (Intermediate B10).

(96) MS ES.sup.+: 476

Intermediate C61: Methyl 4-((trans-N-benzyl-4-(2-methoxyphenoxy)cyclohexanecarboxamido)methyl)benzoate

(97) ##STR00121##

(98) Step (i) A solution of ethyl 4-hydroxycyclohexanecarboxylate (ca. 1:1 mixture of cis and trans ring isomers) (5 g, 29 mmol), 2-methoxyphenol (3.96 g, 31.9 mmol), triphenylphosphine (8.38 g, 31.9 mmol) and trans-diisopropyl diazene-1,2-dicarboxylate (DIAD) (6.21 mL, 31.9 mmol) in THF (150 mL) was stirred at RT. The mixture was diluted with EtOAc (100 mL) and washed with saturated sodium hydrogencarbonate solution (50 mL) and brine (50 mL). The organic phase was dried and concentrated, and the residue purified by silica chromatography (0-100% EtOAc/petrol) to give ethyl 4-(2-methoxyphenoxy)cyclohexanecarboxylate (ca. 1:1 mixture of cis and trans ring isomers) (crude, 4.0 g) which was used without further purification or characterisation.

(99) Step (ii) 4-(2-Methoxyphenoxy)cyclohexanecarboxylic acid was prepared as described for 4-(2-fluorophenoxy)cyclohexanecarboxylic acid (Intermediate C60, step (iii)) using ethyl 4-(2-methoxyphenoxy)cyclohexanecarboxylate from step (i) above.

(100) MS ES.sup.+: 249

(101) Step (iii) The title compound was prepared using the HATU coupling procedure described in Method E4 above from 4-(2-methoxyphenoxy)cyclohexanecarboxylic acid (step (ii) above) and methyl 4-((benzylamino)methyl)benzoate hydrochloride (Intermediate B10).

(102) MS ES.sup.+: 488

Intermediate C62: Methyl 4-((cis-N-(3-(3-fluorophenyl)propyl)-4-(2-methoxyphenoxy)cyclohexanecarboxamido)methyl)benzoate and

Intermediate C63: Methyl 4-((trans-N-(3-(3-fluorophenyl)propyl)-4-(2-methoxyphenoxy)cyclohexanecarboxamido)methyl)benzoate

(103) ##STR00122##

(104) The title compounds were prepared using the HATU coupling procedure described in Method E4 above, from 4-(2-methoxyphenoxy)cyclohexanecarboxylic acid (ca. 1:1 mixture of cis and trans ring isomers) (Intermediate C61, step (ii)) and methyl 4-(((3-(3-fluorophenyl)propyl)amino)methyl)benzoate hydrochloride (Intermediate B18), and separated by preparative HPLC.

(105) MS ES.sup.+: 534 in both cases

Intermediate C64: Methyl 2-fluoro-4-((4-(2-fluorophenoxy)-N-(3-methoxybenzyl benzamido)methyl)benzoate

(106) ##STR00123##

(107) Step (i) Potassium carbonate (3.70 g, 26.7 mmol) was added to a solution of 4-cyano-2-fluorobenzoic acid (4 g, 24.3 mmol) in DMF (350 mL). After stirring at RT for 15 minutes, iodomethane (1.66 mL, 26.7 mmol) was added. The flask was stoppered, and the mixture stirred at 40 C. for 2 h. The mixture was concentrated under reduced pressure and the residue partitioned between DCM (50 mL) and brine (50 mL). The organic phase was passed through silica and concentrated to give methyl 4-cyano-2-fluorobenzoate (4.1 g, 94%).

(108) .sup.1H NMR (300 MHz, CDCl.sub.3) ppm 3.96 (s, 3H) 7.44-7.48 (m, 1H) 7.49-7.53 (m, 1H) 8.01-8.05 (m, 1H)

(109) Step (ii) Raney nickel (2.0 g) was added to a solution of methyl 4-cyano-2-fluorobenzoate (obtained as described in step (i) above) (10.1 g, 61.6 mmol) in acetic acid (200 mL) and water (100 mL). The mixture was stirred at RT under argon at 20 bar. After 18 h the mixture was filtered through diatomaceous earth, washing through with water (1000 mL). The filtrate was extracted with EtOAc (3300 mL). The combined organic phases were dried and concentrated, adding toluene to assist removal of acetic acid, to give methyl 2-fluoro-4-(hydroxymethyl)benzoate (crude, 6.06 g) which was used in the next step without further purification or characterisation.

(110) Step (iii) Methyl 2-fluoro-4-(hydroxymethyl)benzoate (step (ii) above) (1 g, 5.4 mmol) was dissolved in chloroform (40 mL) and THF (5 mL). Manganese(IV) oxide (2.36 g, 27 mmol) was added and the mixture stirred at RT for 2 h and then at 60 C. for 1 h. Further manganese(IV) oxide (2.36 g, 27 mmol) was added and heating was continued at 60 C. After 18 h the mixture was filtered through diatomaceous earth to give methyl 2-fluoro-4-formylbenzoate which was used without further purification.

(111) .sup.1H NMR (300 MHz, CDCl.sub.3) ppm 3.95 (s, 3H) 7.58-7.62 (m, 1H) 7.63-7.67 (m, 1H) 8.08-8.12 (m, 1H) 10.04 (s, 1H)

(112) Step (iv) Methyl 2-fluoro-4-(((3-methoxybenzyl)amino)methyl)benzoate was prepared using the reductive amination procedure described in Method R3 above from (3-methoxyphenyl)methanamine and methyl 2-fluoro-4-formylbenzoate (step (iii) above).

(113) MS ES.sup.+: 304

(114) Step (v) Oxalyl chloride (0.30 mL, 3.51 mmol) was added to a solution of 4-(2-fluorophenoxy)benzoic acid (Intermediate A1) (746 mg, 2.34 mmol) in DCM (10 mL) at RT. DMF (1 drop) was added, and the mixture stirred, allowing to warm to RT. After 2 h, the mixture was concentrated. The residue was dissolved in DCM (5 mL) and added to a solution of methyl 2-fluoro-4-(((3-methoxybenzyl)amino)methyl)benzoate (step (iv) above) (708 mg, 23.4 mmol) and triethylamine (0.49 mL, 3.51 mmol). After 2 h the mixture was partitioned between water (30 mL) and DCM (30 mL). The organic phase was washed sequentially with saturated sodium carbonate solution, citric acid (20% in water) and water, and then dried and concentrated. The residue was purified by silica chromatography (10-30% EtOAc/heptane) to give the title compound which was used in the next step without further purification or characterisation.

Intermediate C65: Methyl 4-((N-(cyclopropylmethyl)-2-fluoro-4-(2-methoxyphenoxy)benzamido)methyl)benzoate

(115) ##STR00124##

(116) Prepared using the HATU coupling procedure described in Method E4 above, from 2-fluoro-4-(2-methoxyphenoxy)benzoic acid (Intermediate A8) and methyl 4-(((cyclopropylmethyl)amino)methyl)benzoate hydrochloride (Intermediate B5). In this case, the acid, amine hydrochloride and DIPEA were combined in DMF first, and HATU added last.

(117) MS ES.sup.+: 464

Intermediate C65a: Methyl 4-((4-bromo-N-(cyclopropylmethyl)benzamido)methyl)benzoate

(118) ##STR00125##

(119) Prepared using a coupling procedure analogous to that described in Method E5 above, from 4-bromobenzoyl chloride and methyl 4-(((cyclopropylmethyl)amino)methyl)benzoate hydrochloride (Intermediate B5).

(120) MS ES.sup.+: 402, 404

Intermediate C65b: Methyl 4-((4-bromo-N-propylbenzamido)methyl)benzoate

(121) ##STR00126##

(122) Prepared using a coupling procedure analogous to that described in Method E5 above, from 4-bromobenzoyl chloride and methyl 4-((propylamino)methyl)benzoate hydrochloride (Intermediate B19a).

(123) MS ES.sup.+: 390, 392

Intermediate C66: Methyl 4-((N-(cyclopropylmethyl)-4-(2-fluoro-6-methoxyphenoxy)benzamido)methyl)benzoate

(124) ##STR00127##

(125) A mixture of methyl 4-((4-bromo-N-(cyclopropylmethyl)benzamido)methyl)benzoate (Intermediate C65a) (300 mg (0.75 mmol), 2-fluoro-6-methoxyphenol (212 mg, 1.49 mmol) and copper(I) oxide (107 mg, 0.75 mmol) in 2,4,6-trimethylpyridine (3 mL) was heated under microwave irradiation at 220 C. for 2 h. Further 2-fluoro-6-methoxyphenol (212 mg, 1.49 mmol) was added, and the mixture heated under microwave irradiation at 220 C. for another 1 h. The mixture was diluted with water and EtOAc (20 mL each) and filtered through diatomaceous earth. The phases were separated. The aqueous phase was acidified to pH 3 by addition of HCl (2 M) and extracted with EtOAc (350 mL). The combined organic phases were dried and concentrated. The residue was purified by silica chromatography (0-60% EtOAc/heptane) to give methyl 4-((N-(cyclopropylmethyl)-4-(2-fluoro-6-methoxyphenoxy)benzamido)methyl)benzoate (90 mg, 26%) as a colourless gum. .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.12-0.31 (m, 2H) 0.38-0.61 (m, 2H) 0.77-1.15 (m, 1H) 2.99-3.50 (m, 2H) 3.90 (s, 3H), 3.79 (s, 3H) 4.59-5.09 (m, 2H) 6.68-7.58 (m, 9H) 7.93-8.10 (m, 2H)

Intermediate C67: Methyl 4-((N-(cyclopropylmethyl)-4-(4-fluoro-2-methoxyphenoxy)benzamido)methyl)benzoate

(126) ##STR00128##

(127) Prepared as described above for Intermediate C66 from methyl 4-((4-bromo-N-(cyclopropylmethyl)benzamido)methyl)benzoate (Intermediate C65a) and 4-fluoro-2-methoxyphenol, except that the mixture was heated under microwave irradiation at 220 C. for 1 h, no additional phenol was required, and slight modifications were made to the workup and purification. In this case the crude reaction mixture was poured into HCl (2 M, 30 mL) and the resulting mixture extracted with EtOAc (350 mL). The combined organic phases were washed with a mixture of NaOH (1 M) and brine (ca. 1:1, total 30 mL), dried and concentrated, and the residue was purified by silica chromatography (20-40% EtOAc/heptane).

(128) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.15-0.30 (m, 2H) 0.40-0.59 (m, 2H) 0.78-1.01 (m, 1H) 2.98-3.45 (m, 2H) 3.78 (s, 3H) 3.91 (s, 3H) 4.64-5.05 (m, 2H) 6.58-7.50 (m, 9H) 7.96-8.10 (m, 2H)

Intermediate C67a: Methyl 4-((4-(4-fluoro-2-methoxyphenoxy)-N-propylbenzamido)methyl)benzoate

(129) ##STR00129##

(130) Prepared as described above for Intermediate C66, from methyl 4-((4-bromo-N-propylbenzamido)methyl)benzoate (Intermediate C65b) and 4-fluoro-2-methoxyphenol, except that the mixture was heated thermally at reflux in air for 18 h, the combined organic phases were washed with HCl (1 M), saturated sodium hydrogencarbonate solution and brine (100 mL each), and the residue was purified by silica chromatography (10-20% EtOAc/heptane).

(131) MS ES.sup.+: 452

Intermediate C67b: Methyl 4-((N-(cyclopropylmethyl)-4-(5-fluoro-2-methoxyphenoxy)benzamido)methyl)benzoate

(132) ##STR00130##

(133) Prepared as described above for Intermediate C66, from methyl 4-((4-bromo-N-(cyclopropylmethyl)benzamido)methyl)benzoate (Intermediate C65a) and 5-fluoro-2-methoxyphenol, except that the combined organic phases were washed with HCl (1 M), NaOH (2 M) and brine (25 mL each).

(134) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.10-0.30 (m, 2H) 0.34-0.62 (m, 2H) 0.75-1.20 (m, 1H) 3.00-3.60 (m, 2H) 3.78 (s, 3H) 3.91 (s, 3H) 4.70-5.00 (m, 2H) 6.70-7.01 (m, 5H) 7.20-7.50 (m, 4H) 7.96-8.06 (m, 2H)

Intermediate C67c: Methyl 4-((4-(5-fluoro-2-methoxyphenoxy)-N-propylbenzamido)methyl)benzoate

(135) ##STR00131##

(136) Prepared as described above for Intermediate C66, from methyl 4-((4-bromo-N-propylbenzamido)methyl)benzoate (Intermediate C65b) and 5-fluoro-2-methoxyphenol, except that the mixture was heated under microwave irradiation at 210 C. for 4 h, the combined organic phases were washed with HCl (1 M), saturated sodium hydrogencarbonate solution and brine (50 mL each), the residue was purified by silica chromatography (20-50% EtOAc/heptane), and the material obtained was used in the next step without further purification or characterisation.

Intermediate C67d: Methyl 4-((N-(cyclopropylmethyl)-4-(3-fluoro-2-methoxyphenoxy)benzamido)methyl)benzoate

(137) ##STR00132##

(138) Prepared as described above for Intermediate C66, from methyl 4-((4-bromo-N-(cyclopropylmethyl)benzamido)methyl)benzoate (Intermediate C65a) and 3-fluoro-2-methoxyphenol (prepared as described in Synthetic Communications, 1985, 15(1), 61-69), except that the mixture was heated under microwave irradiation at 210 C. for 5 h, the combined organic phases were washed with HCl (1 M), saturated sodium hydrogencarbonate solution and brine (100 mL each), and the residue was purified by preparative HPLC.

(139) MS ES.sup.+: 464

Intermediate C67e: Methyl 4-((5-bromo-N-(cyclopropylmethyl)picolinamido)methyl)benzoate

(140) ##STR00133##

(141) Prepared using the coupling procedure described in Method E4 above, from 5-bromopicolinic acid and methyl 4-(((cyclopropylmethyl)amino)methyl)benzoate hydrochloride (Intermediate B5), using 1.3 eq HATU and 6 eq DIPEA.

(142) MS ES.sup.+: 403, 405

Intermediate C68: Methyl 4-((N-(cyclopropylmethyl)-5-(2-methoxyphenoxy)picolinamido)methyl)benzoate

(143) ##STR00134##

(144) Prepared as described above for Intermediate C66, from methyl 4-((5-bromo-N-(cyclopropylmethyl)picolinamido)methyl)benzoate (Intermediate C67e) and 2-methoxyphenol, except that 5 eq phenol and 2.5 eq copper(I) oxide were used, the mixture was heated thermally at reflux in air for 40 h, the aqueous phase was adjusted to pH 7, and the residue was purified by silica chromatography (20-40% EtOAc/heptane).

(145) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.10-0.27 (m, 2H) 0.32-0.57 (m, 2H) 0.85-1.35 (m, 1H) 3.27-3.41 (m, 2H) 3.69-3.85 (m, 3H) 3.89 (s, 3H) 4.89-5.05 (m, 2H) 6.87-7.47 (m, 8H) 7.62-7.71 (m, 1H) 7.90-8.06 (m, 2H)

Intermediate C69: Methyl 4-((N-(cyclopropylmethyl)-5-(2-fluorophenoxy)picolinamido)methyl)benzoate

(146) ##STR00135##

(147) Prepared as described above for Intermediate C68, from methyl 4-((5-bromo-N-(cyclopropylmethyl)picolinamido)methyl)benzoate (Intermediate C67e) and 2-fluorophenol, except that 4 eq phenol was used, the mixture was not filtered through diatomaceous earth, and the residue was purified by silica chromatography (40% EtOAc/heptane).

(148) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.50-0.22 (m, 2H) 0.37-0.52 (m, 2H) 0.95-1.10 (m, 1H) 3.28-3.37 (m, 2H) 3.89 (s, 3H) 4.96 (s, 2H) 7.05-7.40 (m, 7H) 7.64-7.74 (m, 1H) 7.91-8.03 (m, 2H), 8.19-8.34 (m, 1H)

Intermediate C70: 5-(2-Fluorophenoxy)pyrimidine-2-carboxylic acid

(149) ##STR00136##

(150) Step (i) Sodium hydride (60% dispersion in mineral oil, 0.258 g, 6.42 mmol) was added to a solution of 2-fluorophenol (0.492 mL, 5.34 mmol) in pyridine (15 mL) in an ice-water bath. Copper(I) bromide was added, and the mixture heated at 100 C. for 5 h. The mixture was concentrated and the residue triturated with EtOAc and purified by silica chromatography (20% EtOAc/heptane) to give 5-(2-fluorophenoxy)pyrimidine-2-carbonitrile (555 mg, 47%).

(151) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 7.20-7.40 (m, 4H) 8.46 (s, 2H)

(152) Step (ii) 5-(2-Fluorophenoxy)pyrimidine-2-carbonitrile, NaOH (2 M, 10 mL) and IMS (2 mL) were combined and heated at 70 C. for 18 h. After concentration to remove IMS, HCl (2 M) was added until the pH was 3, and the mixture was extracted with DCM (50 mL). The organic phase was dried and concentrated to give 5-(2-fluorophenoxy)pyrimidine-2-carboxylic acid (250 mg, 48%) as a white solid, which was used in the next step without further purification or characterisation.

Intermediate C71: 5-(2-Methoxyphenoxy)pyrimidine-2-carboxylic acid

(153) ##STR00137##

(154) Step (i) Prepared as for step (i) of Intermediate C70 to give 5-(2-methoxyphenoxy)pyrimidine-2-carbonitrile (555 mg, 47%).

(155) MS ES.sup.+: 228

(156) Step (ii) Prepared as for step (ii) of Intermediate C70, from 5-(2-methoxyphenoxy)pyrimidine-2-carbonitrile, to give 5-(2-methoxyphenoxy)pyrimidine-2-carboxylic acid.

(157) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 3.77 (s, 3H) 7.03-7.07 (m, 2H) 7.15-7.19 (m, 1H) 7.28-7.34 (m, 1H) 8.44 (s, 2H)

Intermediate C72: Methyl 4-((N-(cyclopropylmethyl)-5-(2-fluorophenoxy)pyrimidine-2-carboxamido)methyl)benzoate

(158) ##STR00138##

(159) Prepared using the coupling procedure described in Method E4 above, from 5-(2-fluorophenoxy)pyrimidine-2-carboxylic acid (Intermediate C70) and methyl 4-(((cyclopropylmethyl)amino)methyl)benzoate hydrochloride (Intermediate B5).

(160) MS ES.sup.+: 436

Intermediate C73: Methyl 4-((N-(cyclopropylmethyl)-5-(2-methoxyphenoxy)pyrimidine-2-carboxamido)methyl)benzoate

(161) ##STR00139##

(162) Prepared using the coupling procedure described in Method E4 above, from 5-(2-methoxyphenoxy)pyrimidine-2-carboxylic acid (Intermediate C71) and methyl 4-(((cyclopropylmethyl)amino)methyl)benzoate hydrochloride (Intermediate B5).

(163) MS ES.sup.+: 448

Intermediate C74: Methyl 4-((5-(2-methoxyphenoxy)-N-propylpyrimidine-2-carboxamido)methyl)benzoate

(164) ##STR00140##

(165) Prepared using the coupling procedure described in Method E4 above, from 5-(2-methoxyphenoxy)pyrimidine-2-carboxylic acid (Intermediate C71) and methyl 4-((propylamino)methyl)benzoate hydrochloride (Intermediate B19a).

(166) MS ES.sup.+: 436

PREPARATION OF EXAMPLES

(167) Compounds derived from the Intermediates C6 to C74 above were prepared according to Method H1 or H2:

(168) Method H1

(169) Lithium hydroxide or lithium hydroxide monohydrate (5 mmol) was added to a solution of the relevant ester (1 mmol) in water (2 mL) and either THF or 1,4-dioxane (4 mL). In some cases, MeOH was added in addition to THF. The mixture was stirred, typically at RT for 18 h, 50 C. for 4 h or 100 C. for 20 minutes. In some cases the reaction was diluted with NaOH (2 M) and extracted with EtOAc or DCM, and the resulting aqueous phase acidified with HCl (2 M) and extracted with EtOAc. In other cases the crude mixture was partitioned between HCl (2 M) and EtOAc. The combined organic phases from the extraction of the acidic aqueous phase were dried and concentrated. In some cases the residue was loaded onto an anion exchange cartridge. After washing with MeCN, the product was eluted with 1 M HCl/MeCN then concentrated. In some cases the material obtained was purified by reverse-phase chromatography on C18 silica (typically eluted with 5-95% MeOH/H.sub.2O with 0.1% NH.sub.4OH). In some cases the crude product was heated under reflux in HCl (4 M), allowed to cool and filtered. In some cases the final product was recrystallised, typically using one or more of the following: water, EtOH, EtOAc, methyl acetate, methyl tert-butyl ether, pentane, heptane.

(170) Method H2

(171) Sodium hydroxide (2 M, 6 mmol) was added to a solution of the relevant ester (2 mmol) in THF (3 mL). In some cases, MeOH was added instead of or in addition to THF. The mixture was stirred at RT for 18 h or heated under microwave irradiation at 100 C. for 5 minutes. The mixture was partitioned between HCl (1 M) and EtOAc (30 mL each). The organic phases were dried and concentrated. The residue was loaded onto an anion exchange cartridge. After washing with MeCN, the product was eluted with 4 M HCl/1,4-dioxane then concentrated.

Example 1: 4-((N-Ethyl-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid

(172) ##STR00141##

(173) Prepared from the parent methyl ester (Intermediate C6) using Method H1.

(174) .sup.1H NMR (300 MHz, DMSO-d.sub.6) ppm 0.84-1.36 (m, 3H) 3.05-3.64 (m, 2H) 4.67 (br s, 2H) 6.73-7.09 (m, 2H) 7.09-7.61 (m, 8H) 7.74-8.05 (m, 2H) 12.90 (br s, 1H)

(175) MS ES.sup.+: 394

Example 2: 4-((N-(2,2-Difluoroethyl)-4-(2-fluorophenoxy)benzamido)-methyl)benzoic acid

(176) ##STR00142##

(177) Prepared from the parent methyl ester (Intermediate C7) using Method H1.

(178) .sup.1H NMR (300 MHz, DMSO-d.sub.6) ppm 3.57-3.91 (m, 2H) 4.71 (br s, 2H) 5.95-6.52 (m, 1H) 6.82-7.08 (m, 2H) 7.10-7.57 (m, 8H) 7.79-7.98 (m, 2H) 12.93 (br s, 1H)

(179) MS ES.sup.+: 430

Example 3: 4-((4-(2-Fluorophenoxy)-N-(2,2,2-trifluoroethyl)benzamido)-methyl)benzoic acid

(180) ##STR00143##

(181) Prepared from the parent methyl ester (Intermediate C8) using Method H1.

(182) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.14-4.36 (m, 2H) 4.76 (s, 2H) 6.92-7.10 (m, 2H) 7.16-7.37 (m, 5H) 7.39-7.55 (m, 3H) 7.84-7.99 (m, 2H) 12.94 (br s, 1H)

(183) MS ES.sup.+: 448

Example 4: 4-((4-(2-Fluorophenoxy)-N-isobutylbenzamido)methyl)benzoic acid

(184) ##STR00144##

(185) Prepared from the parent methyl ester (Intermediate C9) using Method H1.

(186) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.50-1.08 (m, 6H) 1.77-2.17 (m, 1H) 3.00-3.25 (m, 2H) 4.49-4.86 (m, 2H) 6.88-7.13 (m, 2H) 7.13-7.57 (m, 8H) 7.83-8.04 (m, 2H) 12.69-13.02 (m, 1H)

(187) MS ES.sup.+: 422

Example 5: 4-((N-(Cyclopropylmethyl)-4-(2-fluorophenoxy)benzamido)-methyl)benzoic acid

(188) ##STR00145##

(189) Prepared from the parent methyl ester (Intermediate C10) using Method H1.

(190) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.13-0.26 (m, 2H) 0.30-0.50 (m, 2H) 0.75-1.06 (m, 1H) 2.97-3.22 (m, 2H) 4.79 (br s, 2H) 6.88-7.12 (m, 2H) 7.12-7.55 (m, 8H) 7.86-7.97 (m, 2H) 12.87 (br s, 1H)

(191) MS ES.sup.+: 420

Example 6: 4-((N-(Cyclobutylmethyl)-4-(2-fluorophenoxy)benzamido)-methyl)benzoic acid

(192) ##STR00146##

(193) Prepared from the parent methyl ester (Intermediate C11) using Method H1.

(194) .sup.1H NMR (400 MHz, CD.sub.3OD) ppm 1.37-2.15 (m, 6H) 2.49-2.82 (m, 1H) 3.34-3.58 (m, 2H) 4.60-4.78 (m, 2H) 6.90-7.04 (m, 2H) 7.14-7.29 (m, 5H) 7.33-7.49 (m, 3H) 7.82-8.03 (m, 2H)

(195) MS ES.sup.+: 434

Example 7: 4-((4-(2-Fluorophenoxy)-N-((3-methoxycyclobutyl)methyl)-benzamido)methyl)benzoic acid (mixture of cis and trans ring isomers)

(196) ##STR00147##

(197) Prepared from the parent methyl ester (Intermediate C58) using Method H1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.64-2.30 (m, 4H) 3.04 (br s, 3H) 3.16-3.70 (m, 4H) 4.65 (br s, 2H) 6.85-7.14 (m, 2H) 7.14-7.59 (m, 8H) 7.82-8.00 (m, 2H)

(198) MS ES.sup.+:464

Example 8: 4-((4-(2-Fluorophenoxy)-N-isopentylbenzamido)methyl)benzoic acid

(199) ##STR00148##

(200) Prepared from the parent methyl ester (Intermediate C12) using Method H1.

(201) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.48-1.00 (m, 6H) 1.39 (br s, 3H) 3.21 (br s, 2H) 4.49-4.84 (m, 2H) 7.01 (br s, 2H) 7.17-7.57 (m, 8H) 7.88-7.97 (m, 2H) 12.90 (br s, 1H)

(202) MS ES.sup.+: 436

Example 9: 4-((N-(2-Cyclopropylethyl)-4-(2-fluorophenoxy)benzamido)-methyl)benzoic acid

(203) ##STR00149##

(204) Prepared from the parent methyl ester (Intermediate C13) using Method H1.

(205) .sup.1H NMR (400 MHz, CD.sub.3OD) ppm 0.22-0.17 (m, 2H) 0.26-0.51 (m, 2H) 0.52-0.79 (m, 1H) 1.35-1.64 (m, 2H) 3.35-3.63 (m, 2H) 4.58-4.79 (m, 2H) 6.90-7.08 (m, 2H) 7.13-7.34 (m, 5H) 7.37-7.54 (m, 3H) 8.06-8.22 (m, 2H)

(206) MS ES.sup.+: 434

Example 10: 4-((N-(2-Cyclobutylethyl)-4-(2-fluorophenoxy)benzamido)-methyl)benzoic acid

(207) ##STR00150##

(208) Prepared from the parent methyl ester (Intermediate C14) using Method H1.

(209) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.24-2.30 (m, 9H) 2.88-3.40 (m, 2H) 4.48-4.82 (m, 2H) 7.01 (br s, 2H) 7.19-7.54 (m, 8H) 7.88-7.96 (m, 2H) 12.90 (br s, 1H)

(210) MS ES.sup.+: 448

Example 11: 4-((N-Benzyl-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid

(211) ##STR00151##

(212) Prepared from the parent methyl ester (Intermediate C15) using Method H1.

(213) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.37-4.80 (m, 4H) 6.92-7.06 (m, 2H) 7.11-7.60 (m, 13H) 7.85-7.99 (m, 2H)

(214) MS ES.sup.+: 456

Example 12: 4-((N-(2-Fluorobenzyl)-4-(2-fluorophenoxy)benzamido)-methyl)benzoic acid

(215) ##STR00152##

(216) Prepared from the parent methyl ester (Intermediate C16) using Method H1.

(217) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.45-4.66 (m, 4H) 6.96-7.03 (m, 2H) 7.03-7.45 (m, 10H) 7.45-7.54 (m, 2H) 7.78-7.89 (m, 2H)

(218) MS ES.sup.+: 474

Example 13: 4-((4-(2-Fluorophenoxy)-N-(3-methoxybenzyl)benzamido)-methyl)benzoic acid

(219) ##STR00153##

(220) Prepared from the parent methyl ester (Intermediate C17) using Method H1.

(221) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 3.72 (s, 3H) 4.42-4.71 (m, 4H) 6.55-7.10 (m, 5H) 7.16-7.62 (m, 9H) 7.76-7.99 (m, 2H) 12.92 (br s, 1H)

(222) MS ES.sup.+: 486

Example 14: 4-((4-(2-Fluorophenoxy)-N-(4-methoxybenzyl)benzamido)-methyl)benzoic acid

(223) ##STR00154##

(224) Prepared from the parent methyl ester (Intermediate C18) using Method H1.

(225) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 3.74 (s, 3H) 4.24-4.80 (m, 4H) 6.83-7.65 (m, 14H) 7.84-8.05 (m, 2H) 12.90 (br s, 1H)

(226) MS ES.sup.+: 486

Example 15: 4-((4-(2-Fluorophenoxy)-N-phenethylbenzamido)methyl)benzoic acid

(227) ##STR00155##

(228) Prepared from the parent methyl ester (Intermediate C19) using Method H1.

(229) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 2.40-3.07 (m, 2H) 3.07-4.11 (m, 2H) 4.11-4.95 (m, 2H) 6.55-7.49 (m, 15H) 7.86-8.05 (m, 2H)

(230) MS ES.sup.+: 470

Example 16: 4-((4-(2-Fluorophenoxy)-N-(3-(2-fluorophenyl)propyl)benzamido)methyl)benzoic acid

(231) ##STR00156##

(232) Prepared from the parent methyl ester (Intermediate C20) using Method H1.

(233) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.66-1.98 (m, 2H) 2.17-2.80 (m, 2H) 3.05-3.52 (m, 2H) 4.44-4.90 (m, 2H) 6.83-7.55 (m, 14H) 7.85-8.01 (m, 2H) 12.90 (br s, 1H)

(234) MS ES.sup.+: 502

Example 17: 4-((4-(2-Fluorophenoxy)-N-(3-(3-fluorophenyl)propyl)benzamido)methyl)benzoic acid

(235) ##STR00157##

(236) Prepared from the parent methyl ester (Intermediate C21) using Method H1.

(237) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.70-1.95 (m, 2H) 2.30-2.40 (m, 2H) 3.30-3.40 (m, 2H) 4.74 (br s, 2H) 6.80-7.56 (m, 14H) 7.87-7.99 (m, 2H) 12.88 (br s, 1H)

(238) MS ES.sup.+: 502

Example 18: 4-((4-(2-Fluorophenoxy)-N-((trans-2-phenylcyclopropyl)methyl)-benzamido)methyl)benzoic acid

(239) ##STR00158##

(240) Prepared from the parent methyl ester (Intermediate C22) using Method H1.

(241) .sup.1H NMR (400 MHz, CD.sub.2Cl.sub.2) ppm 0.92 (br s, 2H) 1.20-1.35 (m, 1H) 1.52-1.78 (m, 1H) 3.42 (s, 2H) 4.75-4.99 (m, 2H) 6.86-7.10 (m, 4H) 7.10-7.29 (m, 6H) 7.29-7.50 (m, 5H) 8.00-8.12 (m, 2H)

(242) MS ES.sup.+: 496

Example 19: (S)-4-((4-(2-Fluorophenoxy)-N-(2-hydroxy-3-phenylpropyl)benzamido)methyl)benzoic acid

(243) ##STR00159##

(244) Prepared from the parent methyl ester (Intermediate C23) using Method H1.

(245) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.60-2.76 (m, 1H) 2.95-3.30 (m, 2H) 3.88-4.17 (m, 1H) 4.65 (br s, 2H) 4.85-4.97 (m, 1H) 5.00-5.29 (m, 1H) 6.85-7.33 (m, 12H) 7.33-7.50 (m, 3H) 7.79-7.90 (m, 2H)

(246) MS ES.sup.+: 500

Example 20: (R)-4-((4-(2-Fluorophenoxy)-N-(2-hydroxy-3-phenylpropyl)-benzamido)methyl)benzoic acid

(247) ##STR00160##

(248) Prepared from the parent methyl ester (Intermediate C24) using Method H1.

(249) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.55-2.79 (m, 1H) 2.92-3.40 (m, 2H) 3.83-4.17 (m, 1H) 4.72 (br s, 2H) 4.85-5.05 (m, 1H) 5.06-5.23 (m, 1H) 6.83-7.53 (m, 15H) 7.84-7.97 (m, 2H)

(250) MS ES.sup.+: 500

Example 21: 4-((4-(2-Fluorophenoxy)-N-(4-phenylbutyl)-benzamido)methyl)benzoic acid

(251) ##STR00161##

(252) Prepared from the parent methyl ester (Intermediate C25) using Method H1.

(253) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.29-1.70 (m, 2H) 2.32-2.84 (m, 2H) 3.10-3.55 (m, 2H) 4.62 (br s, 2H) 6.86-7.46 (m, 15H) 7.77-7.95 (m, 2H) 12.57 (br s, 1H)

(254) MS ES.sup.+: 498

Example 22: 4-((N-(Cyclopropylmethyl)-4-(o-tolyloxy)benzamido)methyl)benzoic acid

(255) ##STR00162##

(256) Prepared from the parent methyl ester (Intermediate C26) using Method H1.

(257) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.25-0.25 (m, 2H) 0.25-0.50 (m, 2H) 0.74-1.11 (m, 1H) 2.13 (s, 3H) 2.87-3.47 (m, 2H) 4.78 (br s, 2H) 6.75-7.62 (m, 10H) 7.83-7.98 (m, 2H) 12.87 (br s, 1H)

(258) MS ES.sup.+: 416

Example 23: 4-((N-(Cyclobutylmethyl)-4-(o-tolyloxy)benzamido)methyl)benzoic acid

(259) ##STR00163##

(260) Prepared from the parent methyl ester (Intermediate C27) using Method H1.

(261) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.23-1.99 (m, 7H) 2.15 (s, 3H) 2.40-2.74 (m, 2H) 4.65 (br s, 2H) 6.76-7.64 (m, 10H) 7.87-8.02 (m, 2H) 12.90 (br s, 1H)

(262) MS ES.sup.+: 430

Example 24: 4-((N-Benzyl-4-(o-tolyloxy)benzamido)methyl)benzoic acid

(263) ##STR00164##

(264) Prepared from the parent methyl ester (Intermediate C28) using Method H1.

(265) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.14 (s, 3H) 4.58 (br s, 4H) 6.62-7.04 (m, 3H) 7.07-7.60 (m, 12H) 7.84-8.00 (m, 2H) 12.90 (br s, 1H)

(266) MS ES.sup.+: 452

Example 25: 4-((N-Phenethyl-4-(o-tolyloxy)benzamido)methyl)benzoic acid

(267) ##STR00165##

(268) Prepared from the parent methyl ester (Intermediate C29) using Method H1.

(269) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.15 (br s, 3H) 2.72-2.98 (m, 2H) 3.34-3.59 (m, 2H) 4.43-4.88 (m, 2H) 6.82-7.02 (m, 4H) 7.28 (br s, 11H) 7.88-7.96 (m, 2H)

(270) MS ES.sup.+: 466

Example 26: 4-((N-Phenethyl-4-(2-(trifluoromethyl)phenoxy)benzamido)-methyl)benzoic acid

(271) ##STR00166##

(272) Prepared from the parent methyl ester (Intermediate C30) using Method H1.

(273) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.71-2.99 (m, 2H) 3.34-3.59 (m, 2H) 4.43-4.87 (m, 2H) 6.89-7.42 (m, 12H) 7.43-7.54 (m, 1H) 7.64-7.73 (m, 1H) 7.77-7.85 (m, 1H) 7.88-7.98 (m, 2H)

(274) MS ES.sup.+: 520

Example 27: 4-((N-Benzyl-4-(2-cyanophenoxy)benzamido)methyl)benzoic acid

(275) ##STR00167##

(276) Prepared from the parent methyl ester (Intermediate C31) using Method H1.

(277) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.40-4.76 (m, 4H) 7.00-7.77 (m, 15H) 7.85-8.00 (m, 2H) 12.97 (br s, 1H)

(278) MS ES.sup.+: 463

Example 28: 4-((N-Ethyl-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid

(279) ##STR00168##

(280) Prepared from the parent methyl ester (Intermediate C32) using Method H1.

(281) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.01-1.11 (m, 3H) 3.22-3.33 (m, 2H) 3.73 (s, 3H) 4.68 (br s, 2H) 6.79-6.88 (m, 2H) 6.96-7.03 (m, 1H) 7.07-7.13 (m, 1H) 7.16-7.28 (m, 2H) 7.40 (br s, 4H) 7.89-7.96 (m, 2H) 12.88 (br s, 1H)

(282) MS ES.sup.+: 406

Example 29: 4-((N-(2,2-Difluoroethyl)-4-(2-methoxyphenoxy)benzamido)-methyl)benzoic acid

(283) ##STR00169##

(284) Prepared from the parent methyl ester (Intermediate C33) using Method H1.

(285) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 3.64-3.86 (m, 5H) 4.74 (br s, 2H) 6.07-6.44 (m, 1H) 6.78-6.92 (m, 2H) 6.96-7.03 (m, 1H) 7.07-7.13 (m, 1H) 7.15-7.46 (m, 6H) 7.87-7.97 (m, 2H) 12.93 (br s, 1H)

(286) MS ES.sup.+: 442

Example 30: 4-((N-Isobutyl-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid

(287) ##STR00170##

(288) Prepared from the parent methyl ester (Intermediate C34) using Method H1.

(289) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.58-0.97 (m, 6H) 1.80-2.13 (m, 1H) 3.06-3.19 (m, 2H) 3.72 (s, 3H) 4.68 (s, 2H) 6.77-6.90 (m, 2H) 6.94-7.05 (m, 1H) 7.05-7.13 (m, 1H) 7.16-7.53 (m, 6H) 7.87-7.97 (m, 2H) 12.91 (br s, 1H)

(290) MS ES.sup.+: 434

Example 31: 4-((N-(Cyclopropylmethyl)-4-(2-methoxyphenoxy)benzamido)-methyl)benzoic acid

(291) ##STR00171##

(292) Prepared from the parent methyl ester (Intermediate C35) using Method H1.

(293) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 0.14-0.20 (m, 2H) 0.30-0.44 (m, 2H) 0.81-1.03 (m, 1H) 3.15 (br s, 2H) 3.70 (s, 3H) 4.79 (br s, 2H) 6.78-6.91 (m, 2H) 6.96-7.03 (m, 1H) 7.08-7.14 (m, 1H) 7.16-7.28 (m, 2H) 7.30-7.50 (m, 4H) 7.87-7.96 (m, 2H) 12.88 (s, 1H)

(294) MS ES.sup.+: 432

Example 32: 4-((4-(2-Methoxyphenoxy)-N-(3-phenylpropyl)benzamido)-methyl)benzoic acid

(295) ##STR00172##

(296) Prepared from the parent methyl ester (Intermediate C36) using Method H1.

(297) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.83 (br s, 2H) 2.40 (br s, 2H) 3.22 (br s, 2H) 3.73 (s, 3H) 4.70 (br s, 2H) 6.77-6.82 (m, 2H) 6.95-7.44 (m, 13H) 7.88-7.93 (m, 2H) 12.89 (s, 1H)

(298) MS ES.sup.+: 496

Example 33: 4-((N-(3-(3-Fluorophenyl)propyl)-4-(2-methoxyphenoxy)-benzamido)methyl)benzoic acid

(299) ##STR00173##

(300) Prepared from the parent methyl ester (Intermediate C37) using Method H1.

(301) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.84 (br s, 2H) 2.42 (br s, 2H) 3.21 (br s, 2H) 3.73 (s, 3H) 4.70 (br s, 2H) 6.76-6.82 (m, 2H) 6.83-7.47 (m, 12H) 7.86-7.94 (m, 2H) 12.88 (br s, 1H)

(302) MS ES.sup.+: 514

Example 34: 4-((N-(Cyclopropylmethyl)-4-(2-ethoxyphenoxy)-benzamido)methyl)benzoic acid

(303) ##STR00174##

(304) Prepared from the parent methyl ester (Intermediate C38) using Method H1.

(305) .sup.1H NMR (300 MHz, DMSO-d.sub.6) ppm 0.24-0.60 (m, 4H) 0.69-1.41 (m, 4H) 2.87-3.31 (m, 2H) 3.80-4.15 (m, 2H) 4.76 (br s, 2H) 6.60-7.57 (m, 10H) 7.75-8.00 (m, 2H) 12.86 (br s, 1H)

(306) MS ES.sup.+: 446

Example 35: 4-((4-(2-Chlorophenoxy)-N-(cyclopropylmethyl)benzamido)-methyl)benzoic acid

(307) ##STR00175##

(308) Prepared from the parent methyl ester (Intermediate C39) using Method H1.

(309) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.30-0.23 (m, 2H) 0.23-0.50 (m, 2H) 0.74-1.12 (m, 1H) 2.81-3.41 (m, 2H) 4.79 (br s, 2H) 6.86-7.04 (m, 2H) 7.12-7.66 (m, 8H) 7.84-7.97 (m, 2H) 12.87 (br s, 1H)

(310) MS ES.sup.+: 436

Example 36: 4-((4-(2-Chlorophenoxy)-N-phenethylbenzamido)methyl)benzoic acid

(311) ##STR00176##

(312) Prepared from the parent methyl ester (Intermediate C40) using Method H1.

(313) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.72-2.95 (m, 2H) 3.35-3.60 (m, 2H) 4.41-4.88 (m, 2H) 6.88-7.01 (m, 3H) 7.14-7.54 (m, 11H) 7.58-7.66 (m, 1H) 7.87-7.98 (m, 2H) 12.89 (br s, 1H)

(314) MS ES.sup.+: 486

Example 37: 4-((N-(Cyclopropylmethyl)-4-(2,6-difluorophenoxy)benzamido)-methyl)benzoic acid

(315) ##STR00177##

(316) Prepared from the parent methyl ester (Intermediate C41) using Method H1.

(317) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.00 (br s, 2H) 0.38 (br s, 2H) 0.92 (br s, 1H) 3.12 (br s, 2H) 4.80 (br s, 2H) 7.01 (br s, 2H) 7.24-7.56 (m, 7H) 7.84-7.99 (m, 2H) 12.88 (br s, 1H)

(318) MS ES.sup.+: 438

Example 38: 4-((N-(2-Cyclopropylethyl)-4-(2,6-difluorophenoxy)benzamido)-methyl)benzoic acid

(319) ##STR00178##

(320) Prepared from the parent methyl ester (Intermediate C42) using Method H1.

(321) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.27-0.10 (m, 2H) 0.18-0.75 (m, 3H) 1.25-1.57 (m, 2H) 3.14-3.47 (m, 2H) 4.52-4.81 (m, 2H) 6.87-7.11 (m, 2H) 7.22-7.56 (m, 7H) 7.84-8.00 (m, 2H) 12.88 (br s, 1H)

(322) MS ES.sup.+: 452

Example 39: 4-((N-Benzyl-4-(2,6-difluorophenoxy)benzamido)methyl)benzoic acid

(323) ##STR00179##

(324) Prepared from the parent methyl ester (Intermediate C43) using Method H1.

(325) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.40-4.68 (m, 4H) 6.95-7.04 (m, 2H) 7.11-7.44 (m, 10H) 7.45-7.55 (m, 2H) 7.86-7.94 (m, 2H)

(326) MS ES.sup.+: 474

Example 40: 4-((N-(Cyclopropylmethyl)-4-(2-fluoro-6-methylphenoxy)-benzamido)methyl)benzoic acid

(327) ##STR00180##

(328) Prepared from the parent methyl ester (Intermediate C44) using Method H1.

(329) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.16-0.24 (m, 2H) 0.26-0.48 (m, 2H) 0.77-1.08 (m, 1H) 2.15 (s, 3H) 2.94-3.32 (m, 2H) 4.78 (br s, 2H) 6.88 (br s, 2H) 7.09-7.28 (m, 3H) 7.42 (br s, 4H) 7.85-7.96 (m, 2H) 12.87 (br s, 1H)

(330) MS ES.sup.+: 434

Example 41: 4-((N-(Cyclobutylmethyl)-4-(2-fluoro-6-methylphenoxy)benzamido)methyl)benzoic acid

(331) ##STR00181##

(332) Prepared from the parent methyl ester (Intermediate C45) using Method H1.

(333) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.36-2.01 (m, 6H) 2.16 (s, 3H) 3.22-3.42 (m, 3H) 4.64 (br s, 2H) 6.80-6.95 (m, 2H) 7.17-7.29 (m, 3H) 7.32-7.56 (m, 4H) 7.81-8.02 (m, 2H) 12.88 (br s, 1H)

(334) MS ES.sup.+: 448

Example 42: 4-((4-(2-Chloro-6-fluorophenoxy)-N-(cyclopropylmethyl)-benzamido)methyl)benzoic acid

(335) ##STR00182##

(336) Prepared from the parent methyl ester (Intermediate C46) using Method H1.

(337) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.00 (br s, 2H) 0.38 (br s, 2H) 0.93 (br s, 1H) 3.13 (br s, 2H) 4.80 (br s, 2H) 6.95 (br s, 2H) 7.24-7.60 (m, 7H) 7.86-7.99 (m, 2H) 12.88 (br s, 1H)

(338) MS ES.sup.+: 454

Example 43: 4-((4-(2-Chloro-6-fluorophenoxy)-N-(2-cyclopropylethyl)-benzamido)methyl)benzoic acid

(339) ##STR00183##

(340) Prepared from the parent methyl ester (Intermediate C47) using Method H1.

(341) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.29-0.77 (m, 5H) 1.23-1.57 (m, 2H) 3.15-3.52 (m, 2H) 4.49-4.84 (m, 2H) 6.95 (m, 2H) 7.21-7.57 (m, 7H) 7.89-7.96 (m, 2H) 12.89 (br s, 1H)

(342) MS ES.sup.+: 468

Example 44: 4-((N-Benzyl-4-(2-chloro-6-fluorophenoxy)benzamido)-methyl)benzoic acid

(343) ##STR00184##

(344) Prepared from the parent methyl ester (Intermediate C48) using Method H1.

(345) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.39-4.70 (m, 4H) 6.87-6.99 (m, 2H) 7.08-7.59 (m, 12H) 7.86-7.95 (m, 2H) 12.97 (br s, 1H)

(346) MS ES.sup.+: 490

Example 45: 4-((4-(2,6-Dimethylphenoxy)-N-isopentylbenzamido)methyl)benzoic acid

(347) ##STR00185##

(348) Prepared from the parent methyl ester (Intermediate C49) using Method H2.

(349) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.39-1.61 (m, 9H) 2.06 (s, 6H) 2.75-3.48 (m, 2H) 4.47-4.86 (m, 2H) 6.62-6.89 (m, 2H) 7.05-7.25 (m, 3H) 7.25-7.58 (m, 4H) 7.85-8.02 (m, 2H) 12.89 (br s, 1H)

(350) MS ES.sup.+: 446

Example 46: 4-((N-Benzyl-4-(2,6-dimethylphenoxy)benzamido)methyl)benzoic acid

(351) ##STR00186##

(352) Prepared from the parent methyl ester (Intermediate C50) using Method H2.

(353) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.04 (s, 6H) 4.35-4.79 (m, 4H) 6.68-6.87 (m, 2H) 7.02-7.58 (m, 12H) 7.82-8.02 (m, 2H) 12.89 (br s, 1H)

(354) MS ES.sup.+: 466

Example 47: 4-((4-(2,6-Dimethylphenoxy)-N-phenethylbenzamido)methyl)benzoic

(355) ##STR00187##

(356) Prepared from the parent methyl ester (Intermediate C51) using Method H2.

(357) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.07 (s, 6H) 2.62-3.04 (m, 2H) 3.10-3.70 (m, 2H) 4.35-4.98 (m, 2H) 6.65-7.63 (m, 14H) 7.85-8.03 (m, 2H)

(358) MS ES.sup.+: 480

Example 48: 4-((4-(3-Methoxyphenoxy)-N-phenethylbenzamido)methyl)benzoic acid

(359) ##STR00188##

(360) Prepared from the parent methyl ester (Intermediate C52) using Method H1.

(361) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.72-2.95 (m, 2H) 3.35-3.61 (m, 2H) 3.74 (s, 3H) 4.40-4.88 (m, 2H) 6.54-6.66 (m, 2H) 6.72-6.80 (m, 1H) 6.90-7.11 (m, 3H) 7.13-7.57 (m, 9H) 7.86-8.01 (m, 2H) 12.87 (br s, 1H)

(362) MS ES.sup.+: 482

Example 49: 4-((4-(4-Methoxyphenoxy)-N-phenethylbenzamido)methyl)benzoic acid

(363) ##STR00189##

(364) Prepared from the parent methyl ester (Intermediate C53) using Method H1.

(365) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.70-2.96 (m, 2H) 3.35-3.57 (m, 2H) 3.76 (s, 3H) 4.40-4.87 (m, 2H) 6.85-7.56 (m, 15H) 7.85-8.00 (m, 2H) 12.90 (br s, 1H)

(366) MS ES.sup.+: 482

Example 50: 4-((4-(3-Chlorophenoxy)-N-phenethylbenzamido)methyl)benzoic acid

(367) ##STR00190##

(368) Prepared from the parent methyl ester (Intermediate C54) using Method H1.

(369) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.73-2.98 (m, 2H) 3.36-3.62 (m, 2H) 4.42-4.90 (m, 2H) 6.87-7.57 (m, 15H) 7.93 (br s, 2H) 12.90 (br s, 1H)

(370) MS ES.sup.+: 486

Example 51: 4-((N-(Cyclopropylmethyl)-4-((2-fluorobenzyl)oxy)benzamido)-methyl)benzoic acid

(371) ##STR00191##

(372) Prepared from the parent methyl ester (Intermediate C55) using Method H1.

(373) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.22-0.25 (m, 2H) 0.26-0.47 (m, 2H) 0.77-1.08 (m, 1H) 2.73-3.45 (m, 2H) 4.77 (br s, 2H) 5.15 (s, 2H) 6.98-7.63 (m, 10H) 7.85-7.97 (m, 2H) 12.84 (br s, 1H)

(374) MS ES.sup.+: 434

Example 52: 4-((N-(Cyclobutylmethyl)-4-((2-fluorobenzyl)oxy)benzamido)-methyl)benzoic acid

(375) ##STR00192##

(376) Prepared from the parent methyl ester (Intermediate C56) using Method H1.

(377) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.20-2.20 (m, 7H) 2.26-2.74 (m, 2H) 4.64 (s, 2H) 5.18 (s, 2H) 6.97-7.66 (m, 10H) 7.83-8.07 (m, 2H) 12.81 (br s, 1H)

(378) MS ES.sup.+: 448

Example 53: 4-((N-Benzyl-4-((2-fluorobenzyl)oxy)benzamido)methyl)benzoic

(379) ##STR00193##

(380) Prepared from the parent methyl ester (Intermediate C57) using Method H1.

(381) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 4.56 (br s, 4H) 5.16 (s, 2H) 7.00-7.61 (m, 15H) 7.87-7.99 (m, 2H) 12.92 (br s, 1H)

(382) MS ES.sup.+: 470

Example 54: 4-((N-(Cyclopropylmethyl)-2-fluoro-4-(2-fluorophenoxy)-benzamido)methyl)benzoic acid

(383) ##STR00194##

(384) Prepared from the parent methyl ester (Intermediate C59) using Method H1.

(385) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.06-0.28 (m, 2H) 0.31-0.52 (m, 2H) 0.73-1.15 (m, 1H) 2.99-3.41 (m, 2H) 4.55-4.97 (m, 2H) 6.73-7.07 (m, 2H) 7.25-7.56 (m, 7H) 7.86-8.02 (m, 2H) 12.91 (br s, 1H)

(386) MS ES.sup.+: 438

Example 55: 4-((N-Benzyl-4-(2-fluorophenoxy)cyclohexanecarboxamido)-methyl)benzoic acid (ca. 1:1 mixture of cis and trans ring isomers)

(387) ##STR00195##

(388) Prepared from the parent methyl ester (Intermediate C60) using Method H1.

(389) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.27-2.13 (m, 8H) 2.61-2.86 (m, 1H) 4.00-4.40 (m, 1H) 4.45-4.73 (m, 4H) 6.88-7.00 (m, 1H) 7.05-7.44 (m, 10H) 7.85-8.00 (m, 2H) 12.90 (br s, 1H)

(390) MS ES.sup.+: 462

Example 56: trans-4-((N-Benzyl-4-(2-methoxyphenoxy)cyclohexanecarboxamido)methyl)benzoic acid

(391) ##STR00196##

(392) Prepared from the parent methyl ester (Intermediate C61) using Method H1.

(393) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.10-2.11 (m, 8H) 2.57-2.78 (m, 1H) 3.73 (s, 3H) 4.11-4.25 (m, 1H) 4.46-4.74 (m, 4H) 6.81-7.04 (m, 4H) 7.15-7.44 (m, 7H) 7.86-8.00 (m, 2H) 12.90 (br s, 1H)

(394) MS ES.sup.+: 474

Example 57: cis-4-((N-(3-(3-Fluorophenyl)propyl)-4-(2-methoxyphenoxy)cyclohexanecarboxamido)methyl)benzoic acid

(395) ##STR00197##

(396) Prepared from the parent methyl ester (Intermediate C62) using Method H1.

(397) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.34-1.98 (m, 10H) 2.39-2.66 (m, 3H) 3.15-3.40 (m, 2H) 3.71-3.87 (m, 3H) 4.37-4.51 (m, 1H) 4.52-4.78 (m, 2H) 6.79-7.12 (m, 7H) 7.19-7.39 (m, 3H) 7.82-8.01 (m, 2H) 12.80 (br s, 1H)

(398) MS ES.sup.+: 520

Example 58: trans-4-((N-(3-(3-Fluorophenyl)propyl)-4-(2-methoxyphenoxy)cyclohexanecarboxamido)methyl)benzoic acid

(399) ##STR00198##

(400) Prepared from the parent methyl ester (Intermediate C63) using Method H1.

(401) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 1.19-2.13 (m, 10H) 2.36-2.65 (m, 3H) 3.15-3.49 (m, 2H) 3.67-3.81 (m, 3H) 4.09-4.24 (m, 1H) 4.51-4.78 (m, 2H) 6.79-7.10 (m, 7H) 7.18-7.40 (m, 3H) 7.81-8.00 (m, 2H) 12.86 (br s, 1H)

(402) MS ES.sup.+: 520

Example 59: 2-Fluoro-4-((4-(2-fluorophenoxy)-N-(3-methoxybenzyl)benzamido)methyl)benzoic acid

(403) ##STR00199##

(404) Prepared from the parent methyl ester (Intermediate C64) using Method H1.

(405) .sup.1H NMR (300 MHz, CDCl.sub.3) ppm 3.79 (s, 3H) 4.34-4.91 (m, 4H) 6.57-7.34 (m, 12H) 7.34-7.61 (m, 2H) 7.90-8.07 (m, 1H)

(406) MS ES.sup.+: 504

Example 60: 4-((4-(2-Fluorophenoxy)-N-propylbenzamido)methyl)benzoic acid

(407) ##STR00200##

(408) Prepared from the parent methyl ester (Intermediate C25a) using Method H1.

(409) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.65-1.06 (m, 3H) 1.48-1.79 (m, 2H) 3.11-3.52 (m, 2H), 4.56-4.85 (m, 2H) 6.87-7.52 (m, 10H) 8.06-8.12 (m, 2H)

(410) MS ES.sup.+: 408

Example 61: 4-((4-(2-Methoxyphenoxy)-N-(2,2,2-trifluoroethyl)benzamido)methyl)benzoic acid

(411) ##STR00201##

(412) Prepared from the parent methyl ester (Intermediate C40a) using Method H1.

(413) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 3.79 (s, 3H) 3.84-4.12 (m, 2H) 4.77-4.95 (m, 2H) 6.85-7.47 (m, 10H) 8.03-8.12 (m, 2H)

(414) MS ES.sup.+: 460

Example 62: 4-((4-(2-Methoxyphenoxy)-N-propylbenzamido)methyl)benzoic acid

(415) ##STR00202##

(416) Prepared from the parent methyl ester (Intermediate C40b) using Method H1.

(417) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.65-1.05 (m, 3H) 1.41-1.78 (m, 2H) 3.12-3.52 (m, 2H), 3.80 (s, 3H) 4.55-4.87 (m, 2H) 6.81-7.51 (m, 10H) 8.04-8.12 (m, 2H)

(418) MS ES.sup.+: 420

Example 63: 4-((N-(2,2-Difluoropropyl)-4-(2-methoxyphenoxy)-benzamido)methyl)benzoic acid

(419) ##STR00203##

(420) Prepared from the parent methyl ester (Intermediate C40c) using Method H1.

(421) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.32-1.84 (m, 3H) 3.45-4.02 (m, 5H) 4.65-5.06 (m, 2H) 6.79-7.50 (m, 10H) 7.99-8.15 (m, 2H)

(422) MS ES.sup.+: 456

Example 64: 4-((4-(2-Methoxyphenoxy)-N-(3,3,3-trifluoropropyl)-benzamido)methyl)benzoic acid

(423) ##STR00204##

(424) Prepared from the parent methyl ester (Intermediate C40d) using Method H1.

(425) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 2.35-2.67 (m, 2H) 3.50-3.69 (m, 5H) 4.58-4.81 (m, 2H) 6.80-7.48 (m, 10 OH) 8.06-8.14 (m, 2H)

(426) MS ES.sup.+: 474

Example 65: 4-((N-Butyl-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid

(427) ##STR00205##

(428) Prepared from the parent methyl ester (Intermediate C40e) using Method H1.

(429) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.67-1.01 (m, 3H) 1.01-1.74 (m, 4H) 3.11-3.56 (m, 2H) 4.56-4.90 (m, 2H) 6.80-7.58 (m, 10H) 8.03-8.14 (m, 2H)

(430) MS ES.sup.+: 434

Example 66: 4-((N-(Cyclopropylmethyl)-2-fluoro-4-(2-methoxyphenoxy)benzamido)methyl)benzoic acid

(431) ##STR00206##

(432) Prepared from the parent methyl ester (Intermediate C65) using Method H1.

(433) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.06-0.29 (m, 2H) 0.30-0.53 (m, 2H) 0.78-1.16 (m, 1H) 2.98-3.45 (m, 2H) 3.77 (s, 3H) 4.57-4.98 (m, 2H) 6.59-6.87 (m, 2H) 6.98-7.51 (m, 7H) 7.84-8.00 (m, 2H) 12.90 (br s, 1H)

(434) MS ES.sup.+: 450

Example 67: 4-((N-(Cyclopropylmethyl)-4-(2-fluoro-6-methoxyphenoxy)benzamido)methyl)benzoic acid

(435) ##STR00207##

(436) Prepared from the parent methyl ester (Intermediate C66) using Method H1.

(437) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.10-0.28 (m, 2H) 0.37-0.58 (m, 2H) 0.78-1.15 (m, 1H) 2.98-3.50 (m, 2H) 3.79 (s, 3H) 4.68-5.05 (m, 2H) 6.69-7.50 (m, 9H) 8.02-8.11 (m, 2H)

(438) MS ES.sup.+: 450

Example 68: 4-((N-(Cyclopropylmethyl)-4-(4-fluoro-2-methoxyphenoxy)benzamido)methyl)benzoic acid

(439) ##STR00208##

(440) Prepared from the parent methyl ester (Intermediate C67) using Method H1.

(441) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.09-0.29 (m, 2H) 0.44-0.58 (m, 2H) 0.78-1.17 (m, 1H) 3.01-3.55 (m, 2H) 3.78 (s, 3H) 4.70-5.05 (m, 2H) 6.70-7.52 (m, 9H) 8.03-8.12 (m, 2H)

(442) MS ES.sup.+: 450

Example 69: 4-((N-(Cyclopropylmethyl)-5-(2-methoxyphenoxy)-picolinamido)methyl)benzoic acid

(443) ##STR00209##

(444) Prepared from the parent methyl ester (Intermediate C68) using Method H1.

(445) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.08-0.68 (m, 4H) 0.85-1.33 (m, 1H) 3.16-3.56 (m, 2H) 3.61-4.04 (m, 3H) 4.78-5.19 (m, 2H) 6.80-8.55 (m, 11H)

(446) MS ES.sup.+: 433

Example 70: 4-((4-(o-Tolyloxy)-N-(2,2,2-trifluoroethyl)benzamido)methyl)benzoic acid

(447) ##STR00210##

(448) Prepared from the parent methyl ester (Intermediate C40f) using Method H1.

(449) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 2.18 (s, 3H) 4.02 (br s, 2H) 4.85 (br s, 2H) 6.81-7.58 (m, 10H) 8.00-8.20 (m, 2H)

(450) MS ES.sup.+: 444

Example 71: 4-((4-(2-Fluorophenoxy)-N-(3,3,3-trifluoropropyl)benzamido)methyl)benzoic acid

(451) ##STR00211##

(452) Prepared from the parent methyl ester (Intermediate C25b) using Method H1.

(453) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 2.35-2.65 (m, 2H) 3.53-3.68 (m, 2H) 4.60-4.78 (m, 2H) 6.88-7.45 (m, 10H) 8.06-8.13 (m, 2H)

(454) MS ES.sup.+: 462

Example 72: 4-((4-(2-Chlorophenoxy)-N-(2,2,2-trifluoroethyl)benzamido)methyl)benzoic acid

(455) ##STR00212##

(456) Prepared from the parent methyl ester (Intermediate C40g) using Method H1.

(457) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 3.81-4.18 (m, 2H) 4.84 (br s, 2H) 6.88-7.57 (m, 10H) 8.01-8.16 (m, 2H)

(458) MS ES.sup.+: 464

Example 73: 4-((N-Ethyl-4-(o-tolyloxy)benzamido)methyl)benzoic acid

(459) ##STR00213##

(460) Prepared from the parent methyl ester (Intermediate C40h) using Method H1.

(461) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.92-1.12 (m, 3H) 2.11 (s, 3H) 3.08-3.40 (m, 2H) 4.46-4.78 (m, 2H) 6.74-7.50 (m, 10H) 7.82-7.93 (m, 2H)

(462) MS ES.sup.+: 390

Example 74: 4-((N-(2,2-Difluoropropyl)-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid

(463) ##STR00214##

(464) Prepared from the parent methyl ester (Intermediate C25c) using Method H1.

(465) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 1.24-1.92 (m, 3H) 3.36-4.08 (m, 2H) 4.57-5.13 (m, 2H) 6.79-7.58 (m, 10H) 7.90-8.20 (m, 2H)

(466) MS ES.sup.+: 444

Example 75: 4-((N-Butyl-4-(2-fluorophenoxy)benzamido)methyl)benzoic acid

(467) ##STR00215##

(468) Prepared from the parent methyl ester (Intermediate C25d) using Method H1.

(469) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.66-1.77 (m, 7H) 3.08-3.57 (m, 2H) 4.49-4.93 (m, 2H) 6.81-7.57 (m, 10H) 8.04-8.18 (m, 2H)

(470) MS ES.sup.+: 422

Example 76: 4-((4-(2-Chlorophenoxy)-N-ethylbenzamido)methyl)benzoic acid

(471) ##STR00216##

(472) Prepared from the parent methyl ester (Intermediate C40i) using Method H1.

(473) .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 0.78-1.23 (m, 3H) 2.98-3.65 (m, 2H) 4.40-4.82 (m, 2H) 6.70-7.70 (m, 10 OH) 7.77-8.01 (m, 2H)

(474) MS ES.sup.+: 410

Example 77: 4-((N-(Cyclopropylmethyl)-5-(2-fluorophenoxy)picolinamido)methyl)benzoic acid

(475) ##STR00217##

(476) Prepared from the parent methyl ester (Intermediate C69) using Method H1.

(477) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.06-0.25 (m, 2H) 0.38-0.56 (m, 2H) 0.77-1.33 (m, 1H) 3.27-3.40 (m, 2H) 4.99 (s, 2H) 7.07-8.44 (m, 11H)

(478) MS ES.sup.+: 421

Example 78: 4-((4-(2-Chlorophenoxy)-N-(2,2-difluoroethyl)benzamido)methyl)benzoic acid

(479) ##STR00218##

(480) Prepared from the parent methyl ester (Intermediate C40j) using Method H1.

(481) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 3.59-3.81 (m, 2H) 4.80 (br s, 2H) 5.05-6.40 (m, 1H) 6.81-7.53 (m, 10H) 8.04-8.17 (m, 2H)

(482) MS ES.sup.+: 446

Example 79: 4-((N-(Cyclopropylmethyl)-4-(5-fluoro-2-methoxyphenoxy)benzamido)methyl)benzoic acid

(483) ##STR00219##

(484) Prepared from the parent methyl ester (Intermediate C67b) using Method H1.

(485) .sup.1H NMR (400 MHz, CDCl.sub.3) 0.12-0.32 (m, 2H) 0.41-0.58 (m, 2H) 0.78-1.15 (m, 1H) 3.02-3.52 (m, 2H) 3.78 (s, 3H) 4.68-5.08 (m, 2H) 6.69-7.08 (m, 5H) 7.19-7.58 (m, 4H) 8.04-8.12 (m, 2H)

(486) MS ES.sup.+: 450

Example 80: 4-((4-(4-Fluoro-2-methoxyphenoxy)-N-propylbenzamido)methyl)benzoic acid

(487) ##STR00220##

(488) Prepared from the parent methyl ester (Intermediate C67a) using Method H1.

(489) .sup.1H NMR (300 MHz, CDCl.sub.3) ppm 0.65-1.08 (m, 3H) 1.47-1.74 (m, 2H) 3.10-3.50 (m, 2H), 3.77 (s, 3H) 4.55-4.98 (m, 2H) 6.58-7.58 (m, 9H) 7.90-8.23 (m, 2H)

(490) MS ES.sup.+: 438

Example 81: 4-((4-(5-Fluoro-2-methoxyphenoxy)-N-propylbenzamido)methyl)benzoic acid

(491) ##STR00221##

(492) Prepared from the parent methyl ester (Intermediate C67c) using Method H1.

(493) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.65-1.12 (m, 3H) 1.40-1.84 (m, 2H) 3.03-3.58 (m, 2H), 3.78 (s, 3H) 4.48-5.01 (m, 2H) 6.67-7.12 (m, 5H) 7.20-7.67 (m, 4H) 8.00-8.22 (m, 2H)

(494) MS ES.sup.+: 438

Example 82: 4-((N-(Cyclopropylmethyl)-4-(3-fluoro-2-methoxyphenoxy)benzamido)methyl)benzoic acid

(495) ##STR00222##

(496) Prepared from the parent methyl ester (Intermediate C67d) using Method H1.

(497) .sup.1H NMR (400 MHz, CDCl.sub.3) 0.09-0.27 (m, 2H) 0.42-0.58 (m, 2H) 0.78-1.15 (m, 1H) 3.03-3.55 (m, 2H) 3.86 (s, 3H) 4.70-5.08 (m, 2H) 6.72-6.87 (m, 1H) 6.87-7.08 (m, 4H) 7.22-7.58 (m, 4H) 8.03-8.12 (m, 2H)

(498) MS ES.sup.+: 450

Example 83: 4-((N-(Cyclopropylmethyl)-5-(2-fluorophenoxy)pyrimidine-2-carboxamido)methyl)benzoic acid

(499) ##STR00223##

(500) Prepared from the parent methyl ester (Intermediate C72) using Method H1.

(501) .sup.1H NMR (400 MHz, CDCl.sub.3) 0.06-0.20 (m, 2H) 0.38-0.57 (m, 2H) 0.91-1.14 (m, 1H) 3.01-3.43 (m, 2H) 4.64-5.06 (m, 2H), 7.12-7.33 (m, 4H) 7.40-7.53 (m, 2H) 8.01-8.10 (m, 2H) 8.38-8.57 (m, 2H)

(502) MS ES.sup.+: 422

Example 84: 4-((N-(Cyclopropylmethyl)-5-(2-methoxyphenoxy)pyrimidine-2-carboxamido)methyl)benzoic acid

(503) ##STR00224##

(504) Prepared from the parent methyl ester (Intermediate C73) using Method H1.

(505) .sup.1H NMR (300 MHz, DMSO-d.sub.6) 0.12-0.20 (m, 2H) 0.26-0.42 (m, 2H) 0.76-1.12 (m, 1H) 2.82-3.30 (m, 2H) 3.68-3.76 (m, 3H) 4.35-4.79 (m, 2H), 6.95-7.34 (m, 6H) 7.69-7.82 (m, 2H) 8.39-8.49 (m, 2H)

(506) MS ES.sup.+: 434

Example 85: 4-((5-(2-Methoxyphenoxy)-N-propylpyrimidine-2-carboxamido)methyl)benzoic acid

(507) ##STR00225##

(508) Prepared from the parent methyl ester (Intermediate C74) using Method H1.

(509) .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 0.63-0.99 (m, 3H) 1.48-1.77 (m, 2H) 3.03-3.50 (m, 2H), 3.71-3.86 (m, 3H) 4.49-4.93 (m, 2H) 6.93-7.53 (m, 6H) 7.96-8.14 (m, 2H) 8.28-8.52

(510) MS ES.sup.+: 422

3. Biological Efficacy of Compounds of the Invention

(511) In Vitro LPA5 Receptor Assay

(512) Engagement of the LPA5 receptor by its ligand, oleoyl-L--lysophosphatidic acid (LPA), leads to the release of intracellular stores of calcium into the cytoplasm mediated through a Gq-driven increase in inositol triphosphate (IP3) levels. Gq is a heterotrimeric G protein subunit that activates phospholipase C.

(513) The ability of test compounds to prevent LPA-driven intracellular release of stored calcium from RH7777 cells expressing human LPA5 receptors was assayed by stimulating the cells with LPA in the presence of various test compounds to measure the test compounds' antagonism, i.e. activity in vitro.

(514) Approximately 10,000 cells in normal culture medium (Minimal Essential Media, 10% Foetal Calf Serum) were dispensed per assay well in a black clear bottom poly-D-lysine coated 384 well plate (Corning). Cells were allowed to settle for thirty minutes at room temperature before being incubated overnight at 37 C., 5% CO.sub.2. Sixteen to twenty hours after dispensing, the cells were loaded with a calcium-sensitive fluorescent dye by replacing the culture medium with assay buffer (1 Hanks buffered saline, 25 mM HEPES, 0.1% w/v fatty acid free BSA (bovine serum albumin), pH7.4) containing 1.25 mM probenecid and 1 Calcium 5 Reagent (Molecular Devices). Cells were incubated at 37 C. for one hour to allow for dye uptake.

(515) To test for antagonist activity, test compounds at a final concentration range between 0.32 nM-10 M (diluted in assay buffer) were added to the assay wells and allowed to incubate for fifteen minutes. After incubation with test compounds the assay plate was placed in a FLIPR Tetra system (Molecular Devices) and LPA (diluted in assay buffer) was added to give a final concentration equivalent to its determined EC.sub.90 against LPA5 receptor. Ligand-dependent changes in intracellular calcium levels were determined by measuring changes in fluorescence of the dye over 515-575 nM following excitation over 470-495 nM. Readings from control wells that did not contain test compounds enabled percentage inhibition curves to be plotted using a 4-parameter curve fit algorithm and IC.sub.50 values to be calculated for each compound.

(516) Results

(517) TABLE-US-00005 Example No. Mean IC.sub.50 (nM) Example No. Mean IC.sub.50 (nM) 1 840 2 690 3 43 4 63 5 16 6 24 7 31 8 32 9 15 10 48 11 6 12 9 13 3 14 2 15 120 16 1 17 1 18 2 19 9 20 40 21 11 22 24 23 44 24 7 25 29 26 260 27 200 28 45 29 43 30 12 31 5 32 0.4 33 0.4 34 460 35 7 36 24 37 28 38 140 39 11 40 31 41 89 42 17 43 58 44 11 45 99 46 17 47 11 48 3300 49 4600 50 3600 51 53 52 370 53 58 54 26 55 690 56 180 57 410 58 32 59 6 60 60 61 8 62 9 63 10 64 7 65 4 66 5 67 5 68 10 69 8 70 140 71 83 72 42 73 890 74 98 75 22 76 140 77 74 78 170 79 68 80 45 83 350 84 91 85 120

(518) In Vivo Farnesyl Pyrophosphate-Induced Inflammatory Model

(519) Farnesyl pyrophosphate (FPP) is an endogenous ligand for the LPA5 receptor, with a reported in-vitro EC.sub.50 of 49 nM and 3-fold selectivity over other LPA receptors (Williams et al. (2009), Unique Ligand Selectivity of the GPR92/LPA5 Lysophosphatidate Receptor Indicates Role in Human Platelet Activation, J Biol. Chem., 284(25):17304-17319). The FPP-induced hyperalgesia model is a pharmacodynamic rat model which involves inducing pain by injecting FPP, an agonist of LPA5 receptor, into the plantar surface of the hindpaw. Pre-treatment with a LPA5 receptor antagonist prevents the hyperalgesia induced by FPP.

(520) In adult male rats baseline measurements were taken by assessing the withdrawal threshold to a painful pressure stimulus applied to the hindpaw according to the Randall-Selitto method (Randall and Selitto (1957), A method for measurement of analgesic activity on inflamed tissue, Arch Int Pharmacodyn Ther., 111(4):409-19). The animals were then administered an intraperitoneal (ip) or oral (po) dose of test compound or vehicle 30 to 120 minutes prior to an intra-plantar injection of FPP (100 L at 32 nM) or saline vehicle. Withdrawal responses were reassessed at 30 and 90 minutes after injection of FPP in the ipsilateral paw.

(521) Results

(522) TABLE-US-00006 In vivo efficacy Compound of (minimum effective dose Example No. Route (MED) mg/kg) 3 po 10 4 po 10 4 ip 1 5 po 10 5 ip 0.3 8 po 100 11 po 10 24 po 30 29 po 3 31 po 1 40 po 60

(523) In Vitro LPA1 Receptor Assay

(524) Engagement of the LPA1 receptor by its ligand, oleoyl-L--lysophosphatidic acid (LPA), leads to the release of intracellular stores of calcium into the cytoplasm mediated through a Gq-driven increase in inositol triphosphate (IP3) levels. Gq is a heterotrimeric G protein subunit that activates phospholipase C.

(525) The ability of test compounds to prevent LPA-driven intracellular release of stored calcium from RH7777 cells expressing human LPA1 receptors was assayed by stimulating the cells with LPA in the presence of various test compounds to measure the test compounds' antagonism, i.e. activity in vitro.

(526) Approximately 10,000 cells in normal culture medium (Dulbecco's Modified Eagle Media, 10% Foetal Calf Serum) were dispensed per assay well in a black clear bottom collagen coated 384 well plate (Beckton Dickinson). Cells were allowed to settle for thirty minutes at room temperature before being incubated overnight at 37 C., 5% CO.sub.2. Sixteen to twenty hours after dispensing, the cells were loaded with a calcium-sensitive fluorescent dye by replacing the culture medium with assay buffer (1 Hanks buffered saline, 25 mM HEPES, 0.1% w/v fatty acid free BSA (bovine serum albumin), pH7.4) containing 1.25 mM probenecid and 1 Calcium 5 Reagent (Molecular Devices). Cells were incubated at 37 C. for one hour to allow for dye uptake.

(527) To test for antagonist activity, test compounds at a final concentration range between 0.32 nM-10 M (diluted in assay buffer) were added to the assay wells and allowed to incubate for twenty five minutes. After incubation with test compounds the assay plate was placed in a FLIPR Tetra system (Molecular Devices) and LPA (diluted in assay buffer) was added to give a final concentration equivalent to its determined EC.sub.80 against LPA1 receptor. Ligand-dependent changes in intracellular calcium levels were determined by measuring changes in fluorescence of the dye over 515-575 nM following excitation over 470-495 nM. Readings from control wells that did not contain test compounds enabled percentage inhibition curves to be plotted using a 4-parameter curve fit algorithm and IC.sub.50 values to be calculated for each compound.

(528) TABLE-US-00007 Example No. Mean IC.sub.50 (nM) Example No. Mean IC.sub.50 (nM) 3 35 4 49 5 46 8 46 11 6 13 4 14 2 17 0.4 18 1 19 7 20 12 22 30 24 11 25 14 28 46 29 34 30 12 31 4 32 2 33 0.3 36 24 37 25 38 36 40 160 45 75 46 19 47 22 54 27 58 17 60 33 61 4 62 5 63 7 64 9 65 5 66 7 67 7 68 13 69 12 70 47 71 59 72 29 73 120 74 40 75 31 76 130 77 86 78 390 79 94 80 110 83 540 84 170 85 180

(529) In Vivo Bleomycin Induced Scleroderma Model

(530) Experimental Outline

(531) Adult male C57BL/6 mice, aged 8-10 weeks, were assigned to groups and allowed to acclimatise for a week. On Day 1, the back skin of the animals was shaved. From Day 0, animals were administered with 100 l of a 1 mg/ml solution of bleomycin sulphate in phosphate buffered saline (PBS) by intradermal injection in the shaved skin. Injections were repeated in a one square centimeter area every other day for four weeks. A group of ten animals (Control group) was injected with an identical volume of PBS. Treatments were given according to the administration schedule shown in the table below. Animals were weighed at the beginning of the experiment on Day 0 and then twice weekly throughout the study. At the end of the experiment, on Day 28, animals were culled and samples of back skin were taken for further analysis. All groups were n=10; the vehicle was distilled water and the administration volume was 10 ml/kg for oral gavage (p.o.); administration volume was 25 l for topical application.

(532) TABLE-US-00008 Administration Skin Group Substance Dose Route Frequency sensitisation 1 Vehicle n/a p.o. BID, Day 0-Day PBS, i.d., 28 QAD, Day 0-Day 28 2 Vehicle n/a p.o. BID, Day 0-Day Bleomycin, 28 i.d., QAD, 3 Imatinib 150 mg/kg p.o. BID, Day 0-Day Day 0-Day mesylate 28 28 4 Compound of 30 mg/kg p.o. BID, Day 0-Day Example 5 28 5 Compound of 30 mg/kg p.o. BID, Day 14-Day Example 5 28 6 Protopic 0.1% topical BID, Day 0-Day (trade mark) 28 7 Betamethasone 0.1% topical BID, Day 0-Day 9 SID, Day 10-Day 28

(533) Results

(534) Bodyweights were graphed along with the standard error of the mean (SEM); Skin samples (two punch biopsies per animal, 4 mm in diameter each) samples were hydrolyzed and hydroxyproline content of the samples was determined from analysis in a semi-automated process using a continuous flow analyzer. Autoanalyser readings were calibrated against hydroxyproline standards and collagen content calculated. The bodyweight loss observed in the Example 5 compound-treated groups did not differ from the bodyweight loss observed in the vehicle-treated group. Some weight loss <10% was observed with bleomycin treatment. Bleomycin administration induced a highly significant increase in skin collagen content when compared to the control, saline-injected, group (p<0.01). Imatinib and Betamethasone 0.1% induced a non-significant reduction in skin collagen content when compared to the vehicle-treated group. The Example 5 compound administered from Day 0 and from Day 14 induced a significant reduction in skin collagen content when compared to the vehicle-treated group (p<0.05). Protopic 0.1% induced a highly significant reduction in skin collagen content when compared to the vehicle-treated group (p<0.001). Thus, the compound of Example 5 demonstrated an anti-fibrotic effect in an in vivo bleomycin-induced scleroderma model as measured by collagen skin content.