Isoxazole derivative, preparation method therefor, and use thereof

Abstract

The present invention generally relates to an isoxazole derivative, a preparation therefor, and a use thereof. In particular, the present invention provides a farnesoid X receptor (FXR) agonist compound, and a stereoisomer, a tautomer, a polymorph, a solvate (e.g., a hydrate), a pharmaceutically acceptable salt, an ester, a metabolite, and an N-oxide, and the chemically protected forms and prodrugs thereof. The present invention further provides a preparation method for the compound, an intermediate thereof, and a pharmaceutical composition and kit containing the same and used thereof for treating FXR-mediated diseases or conditions.

Claims

1. A compound of general formula (I) or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, ##STR00088## wherein: A is thiazolylene; B is selected from the group consisting of C.sub.6-10 aryl and 5- to 10-membered heteroaryl containing 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O and S; D is ##STR00089## Z is ##STR00090## W is selected from the group consisting of N and CR.sup.d; R.sup.a is selected from the group consisting of hydrogen, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-8 cycloalkyl, 3- to 14-membered heterocyclyl, C.sub.1-6 alkyl-O—, C.sub.1-6 haloalkyl, C.sub.3-8 halocycloalkyl and C.sub.1-6 haloalkyl-O—; R.sup.b, R.sup.c and R.sup.d are each independently selected from the group consisting of hydrogen, halogen, hydroxyl, cyano, —NH.sub.2, nitro, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.1-6 alkyl-O—, C.sub.1-6 haloalkyl-O—, C.sub.3-8 cycloalkyl, C.sub.3-8 halocycloalkyl, C.sub.3-8 cycloalkyl-O— and C.sub.3-8 halocycloalkyl-O—; R.sup.1 and R.sup.2 are each independently selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, —NH.sub.2, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.1-6 hydroxyalkyl, C.sub.1-6 alkyl-O—, C.sub.1-6 haloalkyl-O—, C.sub.3-8 cycloalkyl, C.sub.3-8 halocycloalkyl, C.sub.1-6 alkyl-NH— and (C.sub.1-6 alkyl).sub.2-N—; R.sup.3a, R.sup.3b, R.sup.3c and R.sup.3d are each independently selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, —NH.sub.2, oxo, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.1-6 hydroxyalkyl, C.sub.1-6 alkyl-O—, C.sub.1-6 haloalkyl-O—, C.sub.3-8 cycloalkyl and C.sub.3-8 halocycloalkyl; alternatively, any two of R.sup.3a, R.sup.3b, R.sup.3c and R.sup.3d together form C.sub.1-6 alkylene; m and n are each independently 0, 1, 2, 3 or 4; and the above alkyl, alkylene, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, hydroxyl, oxo, cyano, —NH.sub.2, nitro, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.1-6 alkyl-O—, C.sub.1-6 haloalkyl-O—, C.sub.3-6 cycloalkyl, C.sub.3-6 halocycloalkyl, C.sub.1-6 alkyl-NH—, (C.sub.1-6 alkyl).sub.2-N—, C.sub.1-6 hydroxyalkyl, cyano-C.sub.1-6 alkyl, 3- to 14-membered heterocyclyl, C.sub.6-14 aryl and 5- to 14-membered heteroaryl.

2. The compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, wherein A is selected from the group consisting of ##STR00091## attached to the ring nitrogen atom in general formula (I) at either of the two positions labeled 1 or 2, and attached to group B at the other position.

3. The compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, wherein B is selected from the group consisting of phenyl and 5- to 6-membered heteroaryl containing 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O and S.

4. The compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, wherein the group ##STR00092## in general formula (I) is ##STR00093##

5. The compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, wherein the compound is a compound of general formula (Ia): ##STR00094##

6. The compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, wherein the compound is selected from the group consisting of: ##STR00095## ##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103## ##STR00104##

7. A pharmaceutical composition comprising at least one compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, and one or more pharmaceutically acceptable carriers.

8. The pharmaceutical composition according to claim 7, which is in a form selected from the group consisting of tablet, capsule, lozenge, hard candy, powder, spray, cream, salve, suppository, gel, paste, lotion, ointment, aqueous suspension, injectable solution, elixir, and syrup.

9. A kit, the kit comprising: a) a first container containing at least one compound according to claim 1, or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, as a first therapeutic agent; b) an optional second container containing at least one additional therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising the additional therapeutic agent as a second pharmaceutical composition; and c) an optional package insert.

10. A method for preparing the compound of general formula (I) according to claim 1, wherein the method comprises the following steps: ##STR00105## wherein: Hal.sup.1, Hal.sup.2 and Hal.sup.3, each independently, are same or different halogens; PG.sup.1 is an amino protecting group; PG.sup.2 is a carboxy protecting group; Y is a boronic acid or borate group; the remaining groups are as defined in claim 1; the reaction conditions for each step are as follows: step A: reacting compound IN-1 with compound IN-2 to obtain compound IN-3; step B: removing the PG.sup.1 group in compound IN-3 to obtain compound IN-4; step C-1: reacting compound IN-4 with compound IN-a to obtain compound IN-5; step D-1: reacting compound IN-5 with compound IN-b to obtain compound IN-6; and step E-1: removing the PG.sup.2 group in compound IN-6 to obtain the compound of general formula (I); alternatively, the method comprises the following steps: ##STR00106## wherein each of the groups is as defined above; the reaction conditions for each step are as follows: step A: reacting compound IN-1 with compound IN-2 to obtain compound IN-3; step B: removing the PG.sup.1 group in compound IN-3 to obtain compound IN-4; step C-2: reacting compound IN-4 with compound IN-c to obtain compound IN-6; and step D-2: removing the PG.sup.2 group in compound IN-6 to obtain the compound of general formula (I).

11. A pharmaceutical composition comprising at least one compound according to claim 5 or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, and one or more pharmaceutically acceptable carriers.

12. A pharmaceutical composition comprising at least one compound according to claim 6 or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, and one or more pharmaceutically acceptable carriers.

13. The pharmaceutical composition according to claim 11, which is in a form selected from the group consisting of tablet, capsule, lozenge, hard candy, powder, spray, cream, salve, suppository, gel, paste, lotion, ointment, aqueous suspension, injectable solution, elixir, and syrup.

14. The pharmaceutical composition according to claim 12, which is in a form selected from the group consisting of tablet, capsule, lozenge, hard candy, powder, spray, cream, salve, suppository, gel, paste, lotion, ointment, aqueous suspension, injectable solution, elixir, and syrup.

15. A kit comprising: a) a first container containing at least one compound according to claim 5 or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, as a first therapeutic agent; b) an optional second container containing at least one additional therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising the additional therapeutic agent as a second pharmaceutical composition; and c) an optional package insert.

16. A kit comprising: a) a first container containing at least one compound according to claim 6 or a stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt, ester, or N-oxide thereof, as a first therapeutic agent; b) an optional second container containing at least one additional therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising the additional therapeutic agent as a second pharmaceutical composition; and c) an optional package insert.

Description

EXAMPLES

(1) The present invention is further described with reference to the following examples, which are not provided to limit the scope of the present invention.

(2) The structure of the compound was confirmed by nuclear magnetic resonance spectrum (.sup.1H NMR) or mass spectrum (MS). .sup.1H-NMR chemical shifts (δ) are expressed in parts per million (ppm). Chemical shifts are expressed in 10.sup.−6 (ppm).

(3) MS was measured using an Agilent (ESI) mass spectrometer.

(4) Preparative high-performance liquid chromatography was conducted on Shimadzu preparative high performance liquid chromatograph.

(5) Thin layer chromatography (TLC) was performed with an aluminum plate (20×20 cm) produced by Merck, and thin-layer chromatography separation and purification employed GF 254 (0.4-0.5 mm).

(6) The reaction was monitored by thin layer chromatography (TLC) or LC-MS, and the developing solvent system included dichloromethane and methanol system, n-hexane and ethyl acetate system, or petroleum ether and ethyl acetate system. It was necessary to adjust the volume ratio of the solvents, or to further add triethylamine, etc. according to the polarity of the compound, in order to achieve the separation and purification of the product.

(7) Microwave reactions were conducted using the Biotage® Initiator+ (400 W, RT˜300° C.) microwave reactor.

(8) Silica gel (200˜300 mesh) was normally employed as a stationary phase in column chromatography. The eluent system included dichloromethane and methanol system, and n-hexane and ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compound; and adjustment by adding a minor amount of triethylamine can be performed.

(9) In the following examples, the reaction temperature was room temperature (20° C.˜30° C.), unless otherwise specified.

(10) The reagents used in this application were purchased from Acros Organics, Aldrich Chemical Company or Shanghai Topbiochem LTD. etc.

(11) In the conventional synthetic methods as well as examples and intermediate preparation examples, abbreviations respectively have the following meanings.

(12) TABLE-US-00001 Abbre- Abbre- viation Meaning viation Meaning TLC Thin layer LC-MS liquid chromatograph- chromatography mass spectrometer Boc tert- BINAP (±)-2,2′- butyloxycarbonyl bis(diphenylphosphino)- Pd(dppf)Cl.sub.2 [1,1′- 1,1′-binaphthalene bis(diphenyl- Pd.sub.2(dba).sub.3 tris(dibenzylidene- phosphino)ferrocene] acetone)dipalladium dichloropalladium DME dimethoxyethane DMF N,N-dimethyl- formamide

Intermediate Preparation Examples

Intermediate Preparation Example 1: Preparation of 4-(((1-(4-bromothiazol-2-yl)piperidin-4-yl)oxy)methyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (T1)

(13) ##STR00032##

Step 1: Preparation of tert-butyl 4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidine-1-carboxylate (T1-b)

(14) N-Boc-4-hydroxypiperidine (1.0 g, 4.95 mmol) was dissolved in dry tetrahydrofuran (20 mL), potassium tert-butoxide (0.54 g, 5.61 mmol) was added, the reaction was stirred for 30 minutes, and 18-crown-6 (1.5 g, 5.61 mmol) and 4-(chloromethyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (T1-a) (1.0 g, 3.31 mmol) were added. The reaction was stirred at room temperature overnight until complete reaction of the starting material indicated by TLC. The mixture was added with water and ethyl acetate. The organic phase was washed with water, dried, concentrated, and the residue was purified by column chromatography on silica gel, to afford the title compound of this step (1.2 g, yield: 78%).

Step 2: Preparation of 5-cyclopropyl-3-(2,6-dichlorophenyl)-4-((piperidin-4-yloxy)methyl)isoxazole (T1-c)

(15) Compound (T1-b) (1.1 g, 2.36 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (5 mL) was added. The reaction was stirred at room temperature for 2 hours until complete reaction of the starting material indicated by TLC. The mixture was concentrated, and the residue was poured to ice water, adjusted to a basic pH with saturated sodium carbonate, and extracted with ethyl acetate. The organic phase was washed with water, dried and concentrated to afford the title compound of this step, which was used directly in the next step without purification.

Step 3: Preparation of 4-(((1-(4-bromothiazol-2-yl)piperidin-4-yl)oxy)methyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (T1)

(16) Compound (T1-c) (500 mg, 1.37 mmol) was dissolved in dry DMF (10 mL), and triethylamine (0.4 mL, 3 mmol) and 2,4-dibromothiazole (398 mg, 1.64 mmol) were added. The mixture was stirred at 80° C. overnight until complete reaction of the starting material indicated by TLC. The mixture was added with water and ethyl acetate, the organic phase was washed with water, dried, concentrated, and the residue was purified by column chromatography on silica gel, to afford the title compound (300 mg, yield: 42%).

Intermediate Preparation Example 2: Preparation of methyl 5-(4-bromophenyl)-1-methyl-1H-pyrazole-3-carboxylate (T2-1) and methyl 3-(4-bromophenyl)-1-methyl-1H-pyrazole-5-carboxylate (T2-2)

(17) ##STR00033##

Step 1: Preparation of methyl 4-(4-bromophenyl)-2,4-dioxobutanoate (T2-b)

(18) 1-(4-bromophenyl)ethanone (T2-a) (1.0 g, 5 mmol) was dissolved in dry tetrahydrofuran (20 mL), sodium hydride (0.24 g, 6 mmol) was added, the reaction was stirred for 30 minutes, and dimethyl oxalate (0.65 g, 5.5 mmol) was added. The reaction was stirred at 70° C. overnight, and cooled. The mixture was added with water and ethyl acetate, the organic phase was washed with water, dried, concentrated, and the residue was purified by column chromatography on silica gel, to afford the title compound of this step (1.2 g, yield: 86%).

Step 2: Preparation of methyl 5-(4-bromophenyl)-1-methyl-1H-pyrazole-3-carboxylate (T2-1) and methyl 3-(4-bromophenyl)-1-methyl-1H-pyrazole-5-carboxylate (T2-2)

(19) Compound (T2-b) (1.4 g, 4.93 mmol) was dissolved in ethanol (30 mL), and methylhydrazine (5 mL) was added. The reaction was stirred at 80° C. overnight, until complete reaction of the starting material indicated by TLC. The mixture was cooled, and then added with water and ethyl acetate. The organic phase was washed with water, dried, concentrated, and the residue was purified by column chromatography on silica gel, to afford compound (T2-1) (300 mg, yield: 20%) and compound (T2-2) (250 mg, yield: 17%).

Intermediate Preparation Example 3: Preparation of methyl 5-(3-bromophenyl)-1-methyl-1H-pyrazole-3-carboxylate (T3-1) and methyl 3-(3-bromophenyl)-1-methyl-1H-pyrazole-5-carboxylate (T3-2)

(20) ##STR00034##

(21) Compound (T3-1) (200 mg, yield: 13%) and compound (T3-2) (230 mg, yield: 15%) were synthesized according to the method described in Intermediate preparation example 2, except that compound (T2-a) was replaced with 1-(3-bromophenyl)ethanone (T3-a) in step 1.

Intermediate Preparation Example 4: Preparation of 4-((((1R,3r,5S)-8-(4-bromothiazol-2-yl)-8-azabicyclo[3.2.1]octan-3-yl)oxy)methyl)-5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazole (T4)

(22) ##STR00035## ##STR00036##

Step 1: Preparation of (E)-2-(trifluoromethoxy)benzaldehyde oxime (T4-b)

(23) 2-(trifluoromethoxy)benzaldehyde (T4-a) (150 g, 788.98 mmol) was dissolved in a mixed solvent of ethanol (1000 mL) and water (1000 mL), hydroxylamine hydrochloride (65.79 g, 946.77 mmol) was added under mechanical stirring, a white solid was precipitated, and a 1 M aqueous solution of sodium hydroxide (789 mL, 788.98 mmol) was further added. The reaction was performed at 25° C. for 2 hours. The mixture was added with 1 M hydrochloric acid (2000 mL) to adjust the pH of the system to about 5, and filtered with suction directly. The solid was dried at 50° C. overnight, to afford compound (T4-b) (150 g, yield: 92%).

Step 2: Preparation of (Z)-N-hydroxy-2-(trifluoromethoxy)benzimidoyl chloride (T4-c)

(24) Compound (T4-b) (150 g, 731.23 mmol) was dissolved in DMF (1000 mL), N-chlorosuccinimide (117.17 g, 877.48 mmol) was added at 0° C. under mechanical stirring, and the reaction was performed at 0° C. for 1 hour. The reactants were added with water (2000 mL), and extracted with ethyl acetate (1000 mL×3). The organic layer was dried over anhydrous sodium sulfate (500 g), filtered and concentrated to afford compound (T4-c) (160 g, yield: 91%).

Step 3: Preparation of methyl 5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazole-4-carboxylate (T4-d)

(25) Methyl 3-cyclopropyl-3-oxopropanoate (189.87 g, 1.34 mol) was added to compound (T4-c) (160 g, 667.84 mmol), the reaction was stirred at −5° C., triethylamine (500 mL) was added dropwise, and the reaction was performed at −5° C. overnight. The reaction solution was poured into water (20 L), and mechanically stirred for 30 minutes, to result in a solid. A yellow solid was obtained by suction filtration, and dried at 50° C. overnight, to afford compound (T4-d) (120 g, yield: 55%).

Step 4: Preparation of (5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methanol (T4-e)

(26) Compound (T4-d) (120 g, 366.69 mmol) was dissolved in toluene, the reaction was stirred at −10° C., 2M diisobutylaluminum hydride (550 mL, 1.10 mol) was added dropwise, and the reaction was performed at room temperature overnight. The reaction solution was poured into methanol (1000 mL) with ice, water (3000 mL) was added under mechanical stirring, and suction filtration was performed to obtain a yellow solid. The filtrate was extracted with ethyl acetate (2000 mL×3), dried over anhydrous sodium sulfate (500 g), filtered, and the organic phase was rotary evaporated to dryness to afford compound (T4-e) (100 g, yield: 91%).

Step 5: Preparation of 4-(chloromethyl)-5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazole (T4-f)

(27) Benzotriazole (59.65 g, 500.74 mmol) was dissolved in dichloromethane, the reaction was stirred at −5° C., and thionyl chloride (59.65 g, 501.39 mmol) was added dropwise. After the reaction was stirred at room temperature for half an hour, a dichloromethane solution (500 mL) of compound (T4-e) (100 g, 334.17 mmol) was added, and the reaction was performed at room temperature for 6 hours. The reaction solution was filtered with suction, and the filtrate was rotary evaporated to dryness to afford compound (T4-f) (106 g, yield: 94%).

Step 6: Preparation of tert-butyl (1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octane-8-carboxylate (T4-g)

(28) Compound (T4-f) (74.77 g, 328.94 mmol) was dissolved in tetrahydrofuran (500 mL), followed by addition of 18-crown-6 (118.56 g, 448.55 mmol). The reaction was stirred at 0° C., potassium tert-butoxide (50.33 g, 448.55 mmol) was added, and then the reaction was stirred at room temperature. Tert-butyl (1R,3r,5S)-3-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylate (95 g, 299.03 mmol) was added, and the reaction was performed at room temperature overnight. The reaction solution was rotary evaporated to dryness, the residue was added with ethyl acetate (1500 mL) and water (1500 mL), the organic phase was washed twice with saturated brine (1500 mL), concentrated and purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−5/1), to afford compound (T4-g) (85 g, yield: 56%).

Step 7: Preparation of 4-(((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yloxy)methyl)-5-cyclopropyl-3-(2-(trifluoromethoxy)phenyflisoxazole (T4-h)

(29) Compound (T4-g) (85 g, 167.15 mmol) was dissolved in a solution of dichloromethane (500 mL), followed by addition of a dioxane solution of hydrochloric acid (4 M, 500 mL). The reaction was performed at room temperature overnight. The reaction solution was rotary evaporated to dryness to afford compound (T4-h) (60 g, yield: 81%).

Step 8: Preparation of 4-((((1R,3r,5S)-8-(4-bromothiazol-2-yl)-8-azabicyclo[3.2.1]octan-3-yl)oxy)methyl)-5-cyclopropyl-3-(2-(trifluoromethoxy)phenyflisoxazole (T4)

(30) Compound (T4-h) (7 g, 15.73 mmol) was dissolved in DMF (50 mL), followed by addition of N,N-diisopropylethylamine (10.17 g, 78.67 mmol) and 2,4-dibromothiazole (4.59 g, 18.88 mmol), and the reaction was performed at 100° C. for 12 hours. The reaction solution was added with water (300 mL), extracted with ethyl acetate (1000 mL), dried over anhydrous sodium sulfate (20 g), and filtered. The organic phase, after being concentrated, was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford compound (T4) (2.8 g, yield: 31%).

Example 1: Preparation of 3-(2-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)thiazol-4-yl)benzoic acid (C1)

(31) ##STR00037##

Step 1: Preparation of methyl 3-(2-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)thiazol-4-yflbenzoate (C1-a)

(32) Compound (T1) (100 mg, 0.19 mmol) and (3-(methoxycarbonyl)phenyl)boronic acid (51 mg, 0.28 mmol) were dissolved in DME (2 mL), a 2N aqueous solution of sodium carbonate (0.14 mL) and Pd(dppf)Cl.sub.2 (13.9 mg, 0.019 mmol) were added, and the reaction was performed at 80° C. for 12 hours. The reactants were then added with water, and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the concentrated residue was purified by column chromatography on silica gel to afford the title compound of this step (50 mg, yield: 45%).

(33) MS m/z (ESI): 584.1 [M+H].sup.+.

(34) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.97-7.92 (m, 4H), 7.66-7.60 (m, 2H), 7.55-7.47 (m, 1H), 7.41 (s, 1H), 4.33 (s, 2H), 3.89 (s, 3H), 3.50-3.39 (m, 2H), 3.29-3.17 (m, 2H), 2.39-2.32 (m, 2H), 1.78-1.69 (m, 2H), 1.44-1.37 (m, 1H), 1.25-1.23 (m, 1H), 1.19-1.05 (m, 4H).

Step 2: Preparation of 3-(2-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)thiazol-4-yl)benzoic acid (C1)

(35) Compound (C1-a) (20 mg, 0.034 mmol) was dissolved in methanol (2 mL) and tetrahydrofuran (1 mL), an aqueous solution of sodium hydroxide (2.74 mg, 0.068 mmol) was added, and the reaction was performed at room temperature for 2 hours. The organic solvents were then removed, the remaining aqueous phase was adjusted to acidic pH with 2N hydrochloric acid, and then extracted with ethyl acetate. The organic phase was concentrated and purified by preparative thin layer chromatography to afford the title compound (15 mg, yield: 78.9%).

(36) MS m/z (ESI): 570.2 [M+H].sup.+.

(37) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 12.91 (s, 1H), 7.97-7.92 (m, 4H), 7.66-7.60 (m, 2H), 7.55-7.47 (m, 1H), 7.41 (s, 1H), 4.33 (s, 2H), 3.50-3.39 (m, 2H), 3.29-3.17 (m, 2H), 2.39-2.32 (m, 2H), 1.78-1.69 (m, 2H), 1.44-1.37 (m, 1H), 1.25-1.23 (m, 1H), 1.19-1.05 (m, 4H).

Example 2: Preparation of 4-(2-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)thiazol-4-yl)benzoic acid (C2)

(38) ##STR00038##

Step 1: Preparation of methyl 4-(2-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)thiazol)-4-yl)benzoate (C2-a)

(39) Compound (C2-a) (45 mg, yield: 40.5%) was synthesized according to the method described in step 1 of Example 1, except that compound (3-(methoxycarbonyl)phenyl)boronic acid was replaced with (4-(methoxycarbonyl)phenyl)boronic acid.

(40) MS m/z (ESI): 584.2 [M+H].sup.+.

(41) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 8.39 (s, 1H), 8.05-8.06 (m, 1H), 7.86-7.84 (m, 1H), 7.62-7.60 (m, 2H), 7.54-7.48 (m, 2H), 7.36 (s, 1H), 4.33 (s, 2H), 3.89 (s, 3H), 3.49-3.39 (m, 3H), 3.25-3.19 (m, 2H), 2.39-2.32 (m, 2H), 1.76-1.67 (m, 2H), 1.45-1.35 (m, 1H), 1.16-1.08 (m, 4H).

Step 2: Preparation of 4-(2-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)thiazol-4-yl)benzoic acid (C2)

(42) The title compound (30 mg, yield: 68.5%) was synthesized according to the method described in step 2 of Example 1, except that compound (C1-a) was replaced with compound (C2-a).

(43) MS m/z (ESI): 570.1 [M+H].sup.+.

(44) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 13.02 (s, 1H), 8.39 (s, 1H), 8.05-8.06 (m, 1H), 7.86-7.84 (m, 1H), 7.62-7.60 (m, 2H), 7.54-7.48 (m, 2H), 7.36 (s, 1H), 4.33 (s, 2H), 3.49-3.39 (m, 3H), 3.25-3.19 (m, 2H), 2.39-2.32 (m, 2H), 1.76-1.67 (m, 2H), 1.45-1.35 (m, 1H), 1.16-1.08 (m, 4H).

Example 3

Preparation of 5-(4-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-3-carboxylic acid (C3)

(45) ##STR00039##

Step 1: Preparation of methyl 5-(4-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-3-carboxylate (C3-a)

(46) Compound (T2-1) (98.8 mg, 0.34 mmol) and compound (T1-c) (103 mg, 0.28 mmol) were dissolved in toluene (2 mL), cesium carbonate (136.9 mg, 0.42 mmol), Pd.sub.2(dba).sub.3 (1.61 mg, 0.0028 mmol) and BINAP (1.74 mg, 0.0028 mmol) were added, and the reaction was performed at 80° C. overnight. The reactants were then added with water, and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography on silica gel to afford the title compound of this step (15 mg, yield: 9.3%).

(47) MS m/z (ESI): 581.2 [M+H].sup.+.

(48) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.61-7.58 (m, 4H), 7.53-7.50 (s, 1H), 6.94 (s, 1H), 6.88-6.86 (m, 2H), 4.30 (s, 2H), 4.06 (s, 3H), 3.87 (s, 3H), 3.27-3.23 (m, 3H), 2.84-2.80 (m, 2H), 2.34-2.32 (m, 1H), 1.72-1.68 (m, 2H), 1.35-1.33 (m, 2H), 1.16-1.09 (m, 4H).

Step 2: Preparation of 5-(4-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-3-carboxylic acid (C3)

(49) Compound (C3-a) (15 mg, 0.026 mmol) was dissolved in methanol (1 mL) and tetrahydrofuran (0.5 mL), an aqueous solution of sodium hydroxide (2.07 mg, 0.52 mmol) was added, and the reaction was performed at room temperature for 2 hours. The organic solvents were then removed, the remaining aqueous phase was adjusted to acidic pH with 2 N hydrochloric acid, and then extracted with ethyl acetate. The organic phase was concentrated, and purified on a preparative thin layer chromatography plate to afford the title compound (2 mg, yield: 13.6%).

(50) MS m/z (ESI): 567.1 [M+H].sup.+.

(51) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 12.92 (s, 1H), 7.61-7.58 (m, 4H), 7.53-7.50 (s, 1H), 6.94 (s, 1H), 6.88-6.86 (m, 2H), 4.30 (s, 2H), 4.06 (s, 3H), 3.27-3.23 (m, 3H), 2.84-2.80 (m, 2H), 2.34-2.32 (m, 1H), 1.72-1.68 (m, 2H), 1.35-1.33 (m, 2H), 1.16-1.09 (m, 4H).

Example 4: Preparation of 3-(4-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-5-carboxylic acid (C4)

(52) ##STR00040##

Step 1: Preparation of methyl 3-(4-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-5-carboxylate (C4-a)

(53) The title compound of this step (30 mg, yield: 15.2%) was synthesized according to the method described in step 1 of Example 3, except that compound (T2-1) was replaced with compound (T2-2).

(54) MS m/z (ESI): 581.2 [M+H].sup.+.

(55) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.62-7.59 (m, 4H), 7.54-7.50 (s, 1H), 6.94 (s, 1H), 6.88-6.86 (m, 2H), 4.31 (s, 2H), 4.11 (s, 3H), 3.87 (s, 3H), 3.27-3.23 (m, 3H), 2.84-2.80 (m, 2H), 2.34-2.32 (m, 1H), 1.72-1.68 (m, 2H), 1.35-1.33 (m, 2H), 1.16-1.09 (m, 4H).

Step 2: Preparation of 3-(4-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-5-carboxylic acid (C4)

(56) The title compound (15 mg, yield: 51%) was synthesized according to the method described in step 2 of Example 3, except that compound (C3-a) was replaced with compound (C4-a).

(57) MS m/z (ESI): 567.1 [M+H].sup.+.

(58) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 12.90 (s, 1H), 7.62-7.59 (m, 4H), 7.54-7.50 (s, 1H), 6.94 (s, 1H), 6.88-6.86 (m, 2H), 4.31 (s, 2H), 4.10 (s, 3H), 3.27-3.23 (m, 3H), 2.84-2.80 (m, 2H), 2.34-2.32 (m, 1H), 1.72-1.68 (m, 2H), 1.35-1.33 (m, 2H), 1.16-1.09 (m, 4H).

Example 5: Preparation of 3-(3-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-5-carboxylic acid (C5)

(59) ##STR00041##

Step 1: Preparation of methyl 3-(3-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-5-carboxylate (C5-a)

(60) The title compound (50 mg, yield: 25.4%) was synthesized according to the method described in step 1 of Example 3, except that compound (T2-1) was replaced with compound (T3-2).

(61) MS m/z (ESI): 581.2 [M+H].sup.+.

(62) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.62-7.59 (m, 2H), 7.53-7.51 (s, 1H), 7.30-7.26 (m, 2H), 7.21-7.19 (m, 2H), 6.87-6.85 (m, 1H), 4.31 (s, 2H), 4.11 (s, 3H), 3.87 (s, 3H), 3.61-3.58 (m, 1H), 3.29-3.25 (m, 2H), 2.86-2.80 (m, 2H), 2.37-2.31 (m, 1H), 1.77-1.70 (m, 3H), 1.16-1.09 (m, 5H).

Step 2: Preparation of 3-(3-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-5-carboxylic acid (C5)

(63) The title compound (40 mg, yield: 82.1%) was synthesized according to the method described in step 2 of Example 3, except that compound (C3-a) was replaced with compound (C5-a).

(64) MS m/z (ESI): 567.1 [M+H].sup.+.

(65) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 12.90 (s, 1H), 7.62-7.59 (m, 2H), 7.53-7.51 (s, 1H), 7.30-7.26 (m, 2H), 7.21-7.19 (m, 2H), 6.87-6.85 (m, 1H), 4.31 (s, 2H), 4.11 (s, 3H), 3.61-3.58 (m, 1H), 3.29-3.25 (m, 2H), 2.86-2.80 (m, 2H), 2.37-2.31 (m, 1H), 1.77-1.70 (m, 3H), 1.16-1.09 (m, 5H).

Example 6: Preparation of 5-(3-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-3-carboxylic acid (C6)

(66) ##STR00042##

Step 1: Preparation of methyl 5-(3-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-3-carboxylate (C6-a)

(67) The title compound of this step (10 mg, yield: 5.1%) was synthesized according to the method described in step 1 of Example 3, except that compound (T2-1) was replaced with compound (T3-1).

(68) MS m/z (ESI): 581.2 [M+H].sup.+.

(69) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.63-7.59 (m, 2H), 7.52-7.51 (s, 1H), 7.30-7.26 (m, 2H), 7.21-7.19 (m, 2H), 6.87-6.85 (m, 1H), 4.31 (s, 2H), 4.08 (s, 3H), 3.89 (s, 3H), 3.61-3.58 (m, 1H), 3.29-3.25 (m, 2H), 2.86-2.80 (m, 2H), 2.37-2.31 (m, 1H), 1.77-1.70 (m, 3H), 1.16-1.09 (m, 5H).

Step 2: Preparation of 5-(3-(4-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-3-carboxylic acid (C6)

(70) The title compound (5 mg, yield: 52.1%) was synthesized according to the method described in step 2 of Example 3, except that compound (C3-a) was replaced with compound (C6-a).

(71) MS m/z (ESI): 567.1 [M+H].sup.+.

(72) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 12.89 (s, 1H), 7.63-7.59 (m, 2H), 7.52-7.51 (s, 1H), 7.30-7.26 (m, 2H), 7.21-7.19 (m, 2H), 6.87-6.85 (m, 1H), 4.31 (s, 2H), 4.08 (s, 3H), 3.61-3.58 (m, 1H), 3.29-3.25 (m, 2H), 2.86-2.80 (m, 2H), 2.37-2.31 (m, 1H), 1.77-1.70 (m, 3H), 1.16-1.09 (m, 5H).

Example 7: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-methylbenzoic acid (C37)

(73) ##STR00043## ##STR00044##

Step 1: Preparation of tert-butyl (1R,3r,5S)-3-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octane-8-carboxylate (C37-a)

(74) At room temperature, compound (T1-a) (1.0 g, 3.33 mmol) was dissolved in DMF (20 mL), after complete dissolution, potassium carbonate (0.919 g, 6.66 mmol) and tert-butyl (1R,3r,5S)-3-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylate (0.756 g, 3.33 mmol) were added under stirring, and the reaction was stirred at 65° C. overnight after the addition. A large number of white solids were precipitated in the reaction solution, and filtered. The filtrate was sequentially washed with water (50 mL) and a saturated brine solution (50 mL). The organic phase was dried over anhydrous sodium sulfate, evaporated under reduced pressure to remove the solvent, and the residue was purified by column chromatography on silica gel to afford the title compound of this step (1.4 g).

(75) MS m/z (ESI): 494.4 [M+H].sup.+.

Step 2: Preparation of 4-(((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yloxy)methyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (C37-b)

(76) Compound (C37-a) (1.4 g, 2.84 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (5 mL) was added, and the reaction was performed at 30° C. for 4 hours. The reaction was monitored by LC-MS until complete reaction of the starting material, and was stopped. The reaction solution was quenched by adding a saturated aqueous solution of sodium bicarbonate (20 mL), and extracted with dichloromethane (30 mL×3). The organic phase was dried over anhydrous sodium sulfate, evaporated under reduced pressure to remove the solvent, in order to afford a crude product, which was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=4/1) to afford the title compound of this step (1.01 g).

(77) MS m/z (ESI): 394.3 [M+H].sup.+.

Step 3: Preparation of 4-((((1R,3r,5S)-8-(4-bromothiazol-2-yl)-8-azabicyclo[3.2.1]octan-3-yl)oxy)methyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (C37-c)

(78) At room temperature, compound (C37-b) (1.01 g, 2.57 mmol) was added to DMF (20 mL), then potassium carbonate (710 mg, 5.14 mmol) and 2,4-dibromothiazole (625 mg, 2.57 mmol) were added, and the reaction was performed at 25° C. overnight until complete reaction of the starting material. The reaction solution was added with water (20 mL), and extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with water, dried over anhydrous sodium sulfate, filtered, and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=5/1) to afford the title compound of this step (830 mg).

(79) MS m/z (ESI): 556.3 [M+H].sup.+.

Step 4: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-methylbenzoate (C37-d)

(80) Compound (C37-c) (830 mg, 1.50 mmol) was added to DMF (20 mL), then methyl 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (414 mg, 1.50 mmol), potassium carbonate (414 mg, 5.14 mmol) and Pd(dppf)Cl.sub.2 (163 mg, 0.2 mmol) were added, and the reaction was performed at 90° C. for 12 hours. TLC indicated the reaction was complete. The reaction was extracted with ethyl acetate (30 mL×3), the combined organic phases were washed with water, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to remove the solvent. The crude product was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=5/1) to afford the title compound of this step (120 mg).

(81) MS m/z (ESI): 625.6 [M+H].sup.+.

Step 5: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-methylbenzoic acid (C37)

(82) At room temperature, compound (C37-d) (120 mg, 0.19 mmol) was added to tetrahydrofuran (10 mL), water (2.0 mL) and lithium hydroxide (14 mg, 0.51 mmol) were then added, and the reaction was performed at room temperature for 6 hours. The reaction continued overnight until complete reaction of the starting material. The reaction solution was adjusted to pH 5-6 with 1 N hydrochloric acid, added with water (100 mL), and then extracted with ethyl acetate (30 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to remove the solvent. The crude product was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=2/1) to afford the title compound (20 mg).

(83) MS m/z (ESI): 610.1 [M+H].sup.+.

(84) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ: 7.90-7.95 (m, 2H), 7.70 (d, J=8.0 Hz, 1H), 7.41-7.36 (m, 3H), 6.59 (s, 1H), 5.30 (s, 2H), 4.26 (s, 2H), 4.16 (s, 2H), 3.52-3.50 (m, 1H), 2.52 (s, 3H), 2.11-2.00 (m, 4H), 1.91-1.68 (m, 6H), 1.25-1.11 (m, 2H).

Example 8: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenypisoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)benzoic acid (C41)

(85) ##STR00045##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)benzoate (C41-a)

(86) Compound (T4) (1 g, 1.75 mmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (532.50 mg, 1.93 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (143.16 mg, 195.66 μmol) and potassium carbonate (483.85 mg, 3.50 mmol) at room temperature. Replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration.

(87) The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (1.12 g).

(88) MS m/z (ESI): 626.1 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)benzoic acid (C41)

(89) Compound (C41-a) (390 mg, 625.31 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution of sodium hydroxide (25.01 mg, 625.31 μmol) (2 mL) was added, and the reaction was performed at 25° C. for 4 hours. The reaction solution was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (150 mg).

(90) MS m/z (ESI): 612.1 [M+H].sup.+.

(91) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.96 (s, 1H), 7.68 (dd, J=18.6, 7.7 Hz, 1H), 7.58 (dd, J=13.9, 6.8 Hz, 1H), 7.45 (s, 1H), 4.36 (s, 1H), 4.15 (s, 1H), 2.36 (s, 1H), 2.04 (d, J=15.2 Hz, 1H), 1.82 (s, 1H), 1.70 (d, J=14.9 Hz, 1H), 1.39-1.26 (m, 1H), 1.13 (d, J=22.2 Hz, 1H).

Example 9: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-methylbenzoic acid (C42)

(92) ##STR00046##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-methylbenzoate (C42-a)

(93) Compound (T4) (500 mg, 876.55 mmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (266.25 mg, 964.20 μmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (71.58 mg, 97.83 μmol) and potassium carbonate (241.93 mg, 1.75 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g) followed by concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (400 mg).

(94) MS m/z (ESI): 640.2 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-methylbenzoic acid (C42)

(95) Compound (C42-a) (300 mg, 468.98 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (18.76 mg, 468.98 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by column chromatography on silica gel (eluent:petroleum ether:ethyl acetate=10:1−6:1) to afford the title compound (280 mg).

(96) MS m/z (ESI): 612.1 [M+H].sup.+.

(97) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.84 (s, 1H), 7.80 (d, J=8.1 Hz, 1H), 7.74-7.68 (m, 1H), 7.66 (d, J=6.2 Hz, 1H), 7.58 (dd, J=13.7, 6.4 Hz, 1H), 7.06 (s, 1H), 4.35 (s, 1H), 4.11 (s, 1H), 2.51 (d, J=15.3 Hz, 4H), 2.41-2.32 (m, 1H), 2.04 (d, J=14.7 Hz, 1H), 1.94 (s, 1H), 1.82 (s, 2H), 1.69 (d, J=14.6 Hz, 1H), 1.19-1.14 (m, 2H), 1.10 (m, 2H).

Example 10: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-fluorobenzoic acid (C43)

(98) ##STR00047##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-fluorobenzoate (C43-a)

(99) Compound (T4) (650 mg, 1.14 mmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (248.13 mg, 1.25 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (93.06 mg, 127.18 μmol) and potassium carbonate (314.50 mg, 2.28 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1) to afford the title compound of this step (350 mg).

(100) MS m/z (ESI): 644.1 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-fluorobenzoic acid (C43)

(101) Compound (C43-a) (390 mg, 625.31 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (25.01 mg, 625.31 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (1.01 g).

(102) MS m/z (ESI): 620.1 [M+H].sup.+.

(103) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.71 (t, J=7.8 Hz, 1H), 7.66 (d, J=7.4 Hz, 1H), 7.63-7.54 (m, 2H), 7.50 (d, J=11.7 Hz, 1H), 7.33 (s, 1H), 4.35 (s, 1H), 4.12 (s, 1H), 2.39-2.33 (m, 1H), 2.03 (d, J=14.5 Hz, 1H), 1.80 (s, 2H), 1.69 (d, J=14.7 Hz, 1H), 1.33-1.24 (m, 1H), 1.16 (m, 1H), 1.10 (m, 1H).

Example 11: Preparation of 4-(2-(1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-2-fluorobenzoic acid (C44)

(104) ##STR00048##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-2-fluorobenzoate (C44-a)

(105) Compound (T4) (400 mg, 701.24 μmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (152.70 mg, 771.36 μmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (57.27 mg, 78.26 μmol) and potassium carbonate (193.54 mg, 1.40 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1) to afford the title compound of this step (200 mg).

(106) MS m/z (ESI): 630.1 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-2-fluorobenzoic acid (C44)

(107) Compound (C44-a) (200.00 mg, 310.73 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (12.43 mg, 310.73 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (170 mg).

(108) MS m/z (ESI): 620.1 [M+H].sup.+.

(109) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.85 (d, J=8.1 Hz, 1H), 7.72 (dd, J=17.5, 10.3 Hz, 1H), 7.66 (d, J=7.3 Hz, 1H), 7.58 (dd, J=13.5, 6.3 Hz, 1H), 7.37 (d, J=1.9 Hz, 1H), 4.36 (s, 1H), 4.15 (s, 1H), 2.37 (d, J=4.9 Hz, 1H), 2.03 (d, J=14.7 Hz, 1H), 1.82 (s, 2H), 1.70 (d, J=14.6 Hz, 1H), 1.33-1.24 (m, 1H), 1.15 (dd, J=11.0, 6.1 Hz, 1H), 1.10 (d, J=3.0 Hz, 1H).

Example 12: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-(trifluoromethyl)benzoic acid (C45)

(110) ##STR00049##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-(trifluoromethyl)benzoate (C45-a)

(111) Compound (T4) (800 mg, 1.40 mmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(trifluoromethyl)benzoate (509.26 mg, 1.54 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (114.53 mg, 156.53 μmol) and potassium carbonate (387.08 mg, 2.80 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1) to afford the title compound of this step (350 mg).

(112) MS m/z (ESI): 693.1 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-3-(trifluoromethyl)benzoic acid (C45)

(113) Compound (C45-a) (350.00 mg, 504.57 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (20.18 mg, 504.57 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (320 mg).

(114) MS m/z (ESI): 680.1 [M+H].sup.+.

(115) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.81 (d, J=8.1 Hz, 1H), 7.70-7.61 (m, 1H), 7.55 (dd, J=12.1, 4.8 Hz, 1H), 7.03 (s, 1H), 4.33 (s, 1H), 4.05 (s, 1H), 2.33 (m), 2.05-1.97 (m, 1H), 1.79 (s, 2H), 1.65 (d, J=14.5 Hz, 1H), 1.25 (d, J=9.3 Hz, 1H), 1.13 (m, 1H), 1.08 (m, 1H).

Example 13: Preparation of 4-(2-(1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-2-(trifluoromethyl)benzoic acid (C46)

(116) ##STR00050##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-2-(trifluoromethyl)benzoate (C46-a)

(117) Compound (T4) (0.4 g, 701.24 μmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)benzoate (277.78 mg, 841.48 μmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (57.27 mg, 70.12 μmol) and potassium carbonate (193.83 mg, 1.40 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=15/1−8/1) to afford the title compound of this step (0.36 g).

(118) MS m/z (ESI): 694.1 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)-2-(trifluoromethyl)benzoic acid (C46)

(119) Compound (C46-a) (360.00 mg, 518.99 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (20.71 mg, 518.99 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (45 mg).

(120) MS m/z (ESI): 680.1 [M+H].sup.+.

(121) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 8.21 (s, 1H), 8.16 (d, J=7.3 Hz, 1H), 7.83 (d, J=7.2 Hz, 1H), 7.64 (dd, J=18.9, 11.7 Hz, 1H), 7.57 (d, J=12.8 Hz, 1H), 4.33 (s, 1H), 4.12 (s, 1H), 2.33 (s, 1H), 2.01 (d, J=12.4 Hz, 1H), 1.79 (s, 1H), 1.68 (m, 1H), 1.19-1.01 (m, 1H).

Example 14: Preparation of 4-(2-(1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-2-carboxylic acid (C47)

(122) ##STR00051##

Step 1: Preparation of methyl 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-2-carboxylate (C47-a)

(123) Compound (T4) (0.5 g, 876.55 μmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiophene-2-carboxylate (282.04 mg, 1.05 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (64.43 mg, 78.89 μmol) and potassium carbonate (241.93 mg, 1.75 mmol) at room temperature, replacement with nitrogen was performed for 2-3 times, and the reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (0.14 g).

(124) MS m/z (ESI): 632.1 [M+H].sup.+.

Step 2: Preparation of 4-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-2-carboxylic acid (C47)

(125) Compound (C47-a) (200.00 mg, 316.61 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (12.66 mg, 316.61 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (0.1 g).

(126) MS m/z (ESI): 618.1 [M+H].sup.+.

(127) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 8.21 (s, 1H), 8.16 (d, J=7.3 Hz, 1H), 7.83 (d, J=7.2 Hz, 1H), 7.64 (dd, J=18.9, 11.7 Hz, 1H), 7.57 (d, J=12.8 Hz, 1H), 4.33 (s, 1H), 4.12 (s, 1H), 2.33 (s, 1H), 2.01 (d, J=12.4 Hz, 1H), 1.79 (s, 1H), 1.68 (m, 1H), 1.19-1.01 (m, 1H).

Example 15: Preparation of 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-3-carboxylic acid (C48)

(128) ##STR00052##

Step 1: Preparation of methyl 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-3-carboxylate (C48-a)

(129) Compound (T4) (0.5 g, 876.55 μmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiophene-3-carboxylate (282.04 mg, 1.05 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (64.43 mg, 78.89 μmol) and potassium carbonate (241.93 mg, 1.75 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (0.2 g).

(130) MS m/z (ESI): 632.1 [M+H].sup.+.

Step 2: Preparation of 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-3-carboxylic acid (C48)

(131) Compound (C48-a) (140.00 mg, 221.63 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (8.86 mg, 221.63 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 ml×3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (70 mg).

(132) MS m/z (ESI): 618.1 [M+H]+.

(133) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 8.14 (s, 1H), 7.74-7.68 (m, 1H), 7.66 (d, J=6.7 Hz, 1H), 7.58 (dd, J=13.4, 6.2 Hz, 1H), 7.26 (s, 1H), 4.35 (s, 1H), 4.09 (s, 1H), 2.36 (s, 1H), 2.01 (d, J=15.4 Hz, 1H), 1.80 (s, 2H), 1.69 (d, J=14.8 Hz, 1H), 1.16 (d, J=8.4 Hz, 1H), 1.10 (d, J=2.8 Hz, 1H).

Example 16: Preparation of 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-2-carboxylic acid (C35)

(134) ##STR00053##

Step 1: Preparation of methyl 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-2-carboxylate (C35-a)

(135) Compound (T4) (709.12 mg, 1.24 mmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiophene-2-carboxylate (0.4 g, 1.49 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (90.87 mg, 111.28 μmol) and potassium carbonate (343.11 mg, 2.48 mmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (0.25 g).

(136) MS m/z (ESI): 632.1 [M+H]+.

Step 2: Preparation of 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)thiophene-2-carboxylic acid (C35)

(137) Compound (C35-a) (200.00 mg, 316.61 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (12.66 mg, 316.61 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, and extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (25 mg).

(138) MS m/z (ESI): 618.1 [M+H].sup.+.

(139) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 7.65 (m, 2H), 7.56 (m, 1H), 7.44 (s, 1H), 7.31 (s, 1H), 4.33 (s, 1H), 4.07 (s, 1H), 3.52 (s, 1H), 2.33 (s, 1H), 1.99 (d, J=14.4 Hz, 1H), 1.78 (s, 2H), 1.66 (d, J=14.6 Hz, 1H), 1.13 (m, 1H), 1.08 (s, 1H).

Example 17: Preparation of 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)picolinic acid (C50)

(140) ##STR00054##

Step 1: Preparation of methyl 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)picolinate (C50-a)

(141) Compound (T4) (50 mg, 87.65 μmol) was dissolved in 1,4-dioxane (20 mL), followed by addition of methyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinate (27.67 mg, 105.19 μmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (7.16 mg, 8.77 μmol) and potassium carbonate (12.11 mg, 87.65 μmol) at room temperature, and replacement with nitrogen was performed for 2-3 times. The reaction was performed at 80° C. for 8 hours. The reaction solution was filtered with suction through celite, the filtrate was washed with ethyl acetate (100 mL×3), and the organic phase thus obtained was dried over anhydrous sodium sulfate (20 g), followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound of this step (20 mg).

(142) MS m/z (ESI): 626.1 [M+H].sup.+.

Step 2: Preparation of 5-(2-((1R,3r,5S)-3-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)-8-azabicyclo[3.2.1]octan-8-yl)thiazol-4-yl)picolinic acid (C50)

(143) Compound (C50-a) (200 mg, 316.61 μmol) was dissolved in tetrahydrofuran (5 mL), an aqueous solution (2 mL) of sodium hydroxide (12.66 mg, 316.61 μmol) was added, and the reaction was performed at 25° C. for 4 hours. The reaction was adjusted to pH 4 with 1 M dilute hydrochloric acid, extracted with ethyl acetate (100 mL×3), and the organic phase was dried over anhydrous sodium sulfate, followed by filtration and concentration. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate=10/1−6/1) to afford the title compound (25 mg).

(144) MS m/z (ESI): 618.1 [M+H].sup.+.

(145) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ: 8.88 (s, 1H), 8.20 (dd, J=8.1, 2.1 Hz, 1H), 7.93 (d, J=8.2 Hz, 1H), 7.71-7.61 (m, 1H), 7.55 (t, J=7.5 Hz, 1H), 7.44 (s, 1H), 4.32 (s, 1H), 4.11 (s, 1H), 3.52 (s, 1H), 2.33 (dd, J=10.8, 5.9 Hz, 1H), 2.01 (d, J=13.7 Hz, 1H), 1.79 (s, 2H), 1.67 (d, J=14.5 Hz, 1H), 1.13 (m, 1H), 1.09-1.03 (m, 1H).

(146) The compounds in following Table 1 were prepared according to synthetic methods similar to those described in above Examples 1-17:

(147) TABLE-US-00002 TABLE 1 Com- pound No. Structure Name MS C7 4-(2-(4-((5-cyclopropyl-3-(2- (trifluoromethoxy)phenyl)isoxazol- 4-yl)methoxy)piperidin-1- yl)thiazol-4-yl)benzoic acid MS m/z (ESI): 586 [M + H].sup.+ C8 4-(2-(4-((5-cyclopropyl-3-(2- (trifluoromethyl)phenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)benzoic acid MS m/z (ESI): 570 [M + H].sup.+ C9 4-(2-(4-((5-cyclopropyl-3-(2,6- difluorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)benzoic acid MS m/z (ESI): 538 [M + H].sup.+ C10 4-(2-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2,6-dichlorophenyl)isoxazol-4- yl)methoxy)-8- azabicyclo[3.2.1]octan-8-yl)thiazol- 4-yl)benzoic acid MS m/z (ESI): 596 [M + H].sup.+ C11 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-5- methylthiazol-4-yl)benzoic acid MS m/z (ESI): 584 [M + H].sup.+ C12 0 6-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)nicotinic acid MS m/z (ESI): 571 [M + H].sup.+ C13 5-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)-2-picolinic acid MS m/z (ESI): 571 [M + H].sup.+ C14 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)-3-methylbenzoic acid MS m/z (ESI): 584 [M + H].sup.+ C15 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)-3-fluorobenzoic acid MS m/z (ESI): 588 [M + H].sup.+ C16 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)-2-methylbenzoic acid MS m/z (ESI): 584 [M + H].sup.+ C17 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)-2-fluorobenzoic acid MS m/z (ESI): 588 [M + H].sup.+ C18 3-(4-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2,6-dichlorophenyl)isoxazol-4- yl)methoxy)-8- azabicyclo[3.2.1]octan-8- yl)phenyl)-1-methyl-1H-pyrazole- 5-carboxylic acid MS m/z (ESI): 593 [M + H].sup.+ C19 3-(6-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1- yl)pyridine-3-yl)-1-methyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 568 [M + H].sup.+ C20 3-(5-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1- yl)pyridine-2-yl)-1-methyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 568 [M + H].sup.+ C21 3-(4-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-3- methylphenyl)-1-methyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 581 [M + H].sup.+ C22 0 3-(4-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-3- fluorophenyl)-1-methyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 585 [M + H].sup.+ C23 3-(4-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-2- methylphenyl)-1-methyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 581 [M + H].sup.+ C24 3-(4-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-2- fluorophenyl)-1-methyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 585 [M + H].sup.+ C25 3-(2-chloro-4-(4-((5-cyclopropyl-3- (2,6-dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)phenyl)- 1-methyl-1H-pyrazole-5-carboxylic acid MS m/z (ESI): 601 [M + H].sup.+ C26 4′-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-[1,1′- biphenyl]-4-carboxylic acid MS m/z (ESI): 563 [M + H].sup.+ C27 4′-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-[1,1′- biphenyl]-3-carboxylic acid MS m/z (ESI): 563 [M + H].sup.+ C28 3-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)-1-methyl-1H-pyrazole-5- carboxylic acid MS m/z (ESI): 574 [M + H].sup.+ C29 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)furan-2-carboxylic acid MS m/z (ESI): 560 [M + H].sup.+ C30 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 4-yl)thiophene-2-carboxylic acid MS m/z (ESI): 576 [M + H].sup.+ C31 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)thiazol- 5-yl)benzoic acid MS m/z (ESI): 570 [M + H].sup.+ C32 0 4-(2-(4-((5-cyclopropyl-3-(2,6- dichlorophenyl)isoxazol-4- yl)methoxy)piperidin-1-yl)-5- methylthiazol-4-yl)-3- methylbenzoic acid MS m/z (ESI): 598 [M + H].sup.+ C33 3-(4-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2- (trifluoromethoxy)phenyl)isoxazol- 4-yl)methoxy)-8- azabicyclo[3.2.1]octan-8- yl)phenyl)-1-methyl-1H-pyrazole- 5-carboxylic acid MS m/z (ESI): 609 [M + H].sup.+ C34 5-(4-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2- (trifluoromethoxy)phenyl)isoxazol- 4-yl)methoxy)-8- azabicyclo[3.2.1]octan-8- yl)phenyl)thiophene-2-carboxylic acid MS m/z (ESI): 611 [M + H].sup.+ C36 4-(2-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2,6-dichlorophenyl)isoxazol-4- yl)methoxy)-8- azabicyclo[3.2.1]octan-8-yl)thiazol- 4-yl)thiophene-2-carboxylic acid MS m/z (ESI): 602 [M + H].sup.+ C38 4-(2-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2,6-dichlorophenyl)isoxazol-4- yl)methoxy)-8- azabicyclo[3.2.1]octan-8-yl)thiazol- 4-yl)-3-fluorobenzoic acid MS m/z (ESI): 614 [M + H].sup.+ C39 5-(2-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2,6-dichlorophenyl)isoxazol-4- yl)methoxy)-8- azabicyclo[3.2.1]octan-8-yl)thiazol- 4-yl)thiophene-3-carboxylic acid MS m/z (ESI): 602 [M + H].sup.+ C40 4-(2-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2,6-dichlorophenyl)isoxazol-4- yl)methoxy)-8- azabicyclo[3.2.1]octan-8-yl)thiazol- 4-yl)-2-methylbenzoic acid MS m/z (ESI): 610 [M + H].sup.+ C49 3-(4-((1R,3r,5S)-3-((5-cyclopropyl- 3-(2- (trifluoromethoxy)phenyl)isoxazol- 4-yl)methoxy)-8- azabicyclo[3.2.1]octan-8- yl)phenyl)-1-isopropyl-1H- pyrazole-5-carboxylic acid MS m/z (ESI): 637 [M + H].sup.+

Biological Assay

Experimental Example 1. Bile Acid Receptor FXR Coactivator Binding Assay

1. Test Method

(148) The Invitrogen⋅LanthaScreen™ TR-FRET Farnesoid X Receptor Coactivator Assay kit was employed to determine the activation effect of the compounds on FXR.

(149) After the receptor was incubated with the test compound at various concentrations at room temperature, the fluorescent-labeled coactivator short peptides and terbium-labeled antibody were added, and the FRET signal was detected after the reaction was complete at room temperature. The group without the receptor protein was employed as a blank, the activation effect (EC.sub.50) and maximum activation effect signal value (Emax) of the test compounds on FXR were calculated according to the four-parameter fitting equation:
y=E min+(E max−E min)/(1+(x/EC.sub.50){circumflex over ( )}(−Hillslope))

(150) wherein y is the FRET binding signal, Emax and Emin respectively are the upper and lower asymptotic estimates of the fitted curve, x is the logarithmic concentration of the compound, and Hillslope is the slope of the curve.

(151) In addition, taking chenodeoxycholic acid (i.e., CDCA) as a positive control, the relative activation effect of the compound tested in the present application was calculated according to the following equation:
Relative activation effect (%)=(E max/E max′)×100%

(152) wherein Emax represents the maximum activation effect value of the compound tested in the present application, and Emax′ represents the maximum activation effect value of CDCA, both of them are calculated according to the equation as described above.

2. Test Results

(153) TABLE-US-00003 TABLE 2 EC.sub.50 of the compounds tested in the present application on FXR Compound No. EC.sub.50 (μM) CDCA 4.43 ± 0.84 C1  0.178 ± 0.021 C2  0.012 ± 0.004 C3  0.042 ± 0.007 C4  0.010 ± 0.001 C6  0.206 ± 0.018 C35 0.013 ± 0.003 C37 0.015 ± 0.001 C44 0.022 ± 0.005 C45 0.110 ± 0.021 C46 0.002 ± 0.001 C50 0.031 ± 0.008

(154) According to the data in Table 2, the compounds tested have lower EC.sub.50 values (0.002-0.206 μM), compared with chenodeoxycholic acid (CDCA) (EC.sub.50 value: 4.43 μM), indicating that the compounds of the present invention have a better activation effect on FXR.

(155) TABLE-US-00004 TABLE 3 Relative activation effect of the compounds tested in the present application on FXR Compound No. Relative activation effect CDCA 100% C2  101% C4  102% C35 152% C37 139% C44 137% C45 113% C46 196% C50 177%

(156) According to the data in Table the maximum activation effect values of the compounds tested in the present application are comparable or higher than that of chenodeoxycholic acid (CDCA), indicating that the compounds of the present invention have a good maximum activation effect on FXR.

(157) Taken together, the EC.sub.50 values and relative activation effect data in Tables 2 and 3 indicate that the compounds of the present invention have a good activation effect on FXR.

(158) The remaining compounds of the present invention also have good EC.sub.50 values and relative activation effects, that is, they have good activation activity and maximum activation effect on FXR.

Experimental Example 2. Luciferase Reporter Gene Assay

1. Test Method

(159) Human embryonic kidney cells HEK293 were cultured in a DMEM medium containing 10% FBS. Plasmids were cotransfected to allow the cells to have over expression of FXR and human BSEP luciferase reporter genes. The transfected cells were digested, resuspended, counted, and plated in a multi-well plate. 10 μL of the test compound at various concentrations was added to the multi-well plate to result in final concentrations of 64 μM, 16 μM, 4 μM, 1 μM, 0.25 μM, 0.0625 μM, 0.0156 μM, 0.0039 μM, 0.000975 μM, 0.000244 μM, 0 μM, and the final concentration of DMSO was 0.5%. After the test compound was incubated with the cells for 18 hours, the Brigh-Glo™ detection reagent was added, and the chemical light unit value (RLU) was detected by a multifunctional automatic microplate reader. Taking the signal value of the blank well (without the test compound) as 100%, the relative signal percentage (%) at each concentration of the test compound was calculated. The SigmaPlot 10 software was employed to fit the EC.sub.50 and the maximum activation effect Emax (relative signal percentage) of the test compounds using a four-parameter model.

2. Test Results

(160) The test results are shown in Table 4 below.

(161) TABLE-US-00005 TABLE 4 Compound No. EC.sub.50 (μM) Emax C2  0.02 ± 0.01 248% C4  0.04 ± 0.01 255% C35 0.091 ± 0.038 378% C37 0.006 ± 0.002 411% C41 0.058 ± 0.023 252% C42 0.033 ± 0.018 320% C43 0.037 ± 0.019 343% C44 0.021 ± 0.006 322% C46 0.014 ± 0.005 301% C47 0.019 ± 0.007 370% C48 0.039 ± 0.017 382%

(162) According to the data in Table 4, in the in vitro cell assay, the compounds tested in the present application have EC.sub.50 values between 0.006 μM and 0.091 μM and Emax values greater than 200%. It is shown that the compounds of the present invention have good FXR activation activity in the in vitro cell assay.

(163) The remaining compounds of the present invention also have good EC.sub.50 and Emax values, that is, they have good FXR activation activity in the in vitro cell assay.

Experimental Example 3: Study on Pharmacokinetics (PK) and Liver Tissue Distribution in Rats

(164) The test compound was administered to male SD rats by intravenous (IV) and by gavage (PO), and the pharmacokinetics and liver tissue distribution characteristics of the test compound were evaluated. The doses of the IV and PO administration were respectively 1 mg/kg and 5 mg/kg. The vehicle for IV was 5% DMSO:5% Solutol:90% physiological saline, and the vehicle for PO was 0.5% MC. Blood and liver were collected at various time points after the IV and PO administration. The blood was treated with EDTA. K.sub.2 for anticoagulation, and was centrifuged to obtain a plasma sample; the liver was homogenized and stored at −80° C. The plasma and liver samples were analyzed by LC-MS/MS after being subjected to protein precipitation.

(165) By employing the WinNonlin 6.3 software, the pharmacokinetic parameters were calculated according to a non-compartment model. The results are shown in Tables 5 and 6.

(166) TABLE-US-00006 TABLE 5 Pharmacokinetic parameters of the test compound administered by IV in rats Compound Administration Dosage AUC.sub.last C.sub.max No. route mg/kg h*ng/mL ng/mL C2 IV 1 2409 ± 108 4343 ± 172

(167) According to the data in Table 5, after IV administration of compound C2 of the present invention at a dose of 1 mg/kg, the AUC.sub.last in rats is 2409±108 h*ng/mL and the C.sub.max is 4343±172 ng/mL, indicating that compound C2 of the present invention administered by IV in rats has good drug exposure.

(168) TABLE-US-00007 TABLE 6 Pharmacokinetic parameters of the test compound administered by PO in rats Compound Administration No. route Dosage AUC.sub.last C.sub.max C2 PO 5 mg/kg 1177 ± 304 252 ± 30 h*ng/mL (blood) ng/mL (blood) C2 PO 5 mg/kg 14941 ± 4276 3367 ± 830 h*ng/g (liver) ng/g (liver)

(169) According to the data in Table 6, after PO administration of compound C2 of the present invention at a dose of 5 mg/kg, the AUC.sub.last in the blood and liver of the rats are respectively 1177±304 h*ng/mL and 14941±4276 h*ng/g, and the C.sub.max in the blood and liver of the rats are respectively 252±30 ng/mL and 3367±830 ng/g, indicating that compound C2 of the present invention administered by PO in rats has certain drug exposure and a significant liver enrichment effect.

(170) Taken together, the data in Tables 5 and 6 indicate the compound of Example 2 of the present invention administered by IV in rats has good plasma drug exposure, and the compound administered by PO in rats has certain drug exposure and significant liver enrichment effect.

(171) The remaining compounds of the present invention also have good AUC.sub.last and C.sub.max values, and thus have good pharmacokinetic properties in rats.

(172) When applied as a drug for a disease mediated by FXR, the compound of the present invention exhibits a good effect in terms of drug safety, and shows good drug activity and in vivo metabolic advantages is respect of pharmacodynamics or pharmacokinetics in animals or in vitro.

Experimental Example 4: Liver Microsomal Stability Test

Test Method

(173) The test compound (50 μL) was mixed with the liver microsome (100 μL) of each species, after pre-incubation at 37° C. for 5 minutes, NADPH (50 μL) was added, and the incubation was performed for 0, 30, and 60 minutes. The incubation concentrations of the test compound, NADPH, and liver microsomal enzyme were 1 μM, 1 mM, and 0.5 mg/mL, respectively. Ice-cold acetonitrile (200 μL) was added to stop the reaction, and an appropriate volume of the internal standard was then added. After the vortex and centrifugation process, the supernatant was obtained and detected.

Detection Method

(174) LC-MS/MS, wherein the mass spectrometer was API 5500, and the liquid chromatograph was Shimadzu LC-30AD system. The chromatographic column was Hypersil GOLD C18, 1.9 μm particle size, 50×2.1 mm; mobile phase A was water+0.1% formic acid, phase B was acetonitrile; flow rate was 0.55 mL/min, and column temperature was 40° C. The ion source was an ESI source positive ion mode, and the scanning mode is multiple reaction monitoring (MRM).

(175) By determining the concentrations of the samples at different incubation time, the rate constant was obtained by plotting “Ln (amount of the remained drug %)” versus “incubation time”. The half-life and liver clearance rate of the drug were then calculated, and were used to evaluate the metabolic stability of the drug in liver microsome.

Test Results

(176) TABLE-US-00008 TABLE 7 Liver microsomal stability test Liver microsomal stability of compound C46 CL.sub.int (liver) Species T.sub.1/2 (min) ml/min/kg rat 963 0.96 mouse 150 15.3 dog 259 8.03

(177) Conclusion: according to the experimental data in Table 7, compound C46 of the present invention has a good clearance rate in liver microsome, and the clearance rate in human liver microsome is 14.9 ml/min/kg, indicating good clearance.

(178) In addition to those embodiments described herein, according to the foregoing description, various modifications to the present invention would be apparent to those skilled in the art. Such modifications are intended to fall within the scope of the appended claims. Each reference cited herein (including all patents, patent applications, journal articles, books and any other disclosures) are incorporated herein by reference in its entirety.