Aromatic compound, pharmaceutical composition and use thereof

Abstract

The invention relates to an aromatic compound, pharmaceutical composition comprising the same, and a method for preparing the compound and an intermediate thereof. The invention also relates to use of the compound for the manufacture of a medicament for the prevention or treatment of a PPAR-related disease.

Claims

1. A compound or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein the compound has the structure of formula (I): ##STR00042## wherein: R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are each independently selected from the group consisting of H, halogen, —OH, —SH, cyano, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, halogenated C.sub.1-6 alkyl, C.sub.2-5 alkenyl, —O—[(C.sub.1-6 alkylene)-O].sub.n—(C.sub.1-6 alkyl), —S—(C.sub.1-6 alkyl), —NH.sub.2, —NH—(C.sub.1-6 alkyl), —N(C.sub.1-6 alkyl).sub.2 and 3-10 membered heterocyclyl; or, R.sup.3 and R.sup.4 are connected to form C.sub.3-6 cycloalkyl or 3-10 membered heterocyclyl; X is selected from ethylene, vinylene and C.sub.3-6 cycloalkylene; optionally, the ethylene, vinylene and C.sub.3-6 cycloalkylene are each independently substituted by one or more substituents selected from the group consisting of halogen, C.sub.1-6 alkyl, halogenated C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, —O—(C.sub.1-6 alkyl), 3-10 membered heterocyclyl, C.sub.6-14 aryl and 5-14 membered heteroaryl; Y is selected from a bond, N and C—R.sup.6; W is selected from N and S; R.sup.5 is selected from the group consisting of H, halogen, —OH, —SH, cyano, nitro, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.2-5 alkenyl, —O—[(C.sub.1-6 alkylene)-O].sub.n—(C.sub.1-6 alkyl), —O—(C.sub.3-6 cycloalkyl), —O-(3-6 membered heterocyclyl), —S(O).sub.m—(C.sub.3-6 cycloalkyl), —S(O).sub.m-(3-10 membered heterocyclyl), —NH.sub.2, —NH—(C.sub.1-6 alkyl), —N(C.sub.1-6 alkyl).sub.2, 3-10 membered heterocyclyl, C.sub.6-14 aryl and 5-14 membered heteroaryl; optionally, the C.sub.1-6 alkyl, cycloalkyl, heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl are each independently substituted by one or more substituents selected from the group consisting of halogen, —OH, —OC.sub.1-6 alkyl, halogenated C.sub.1-6 alkyl, —O-halogenated C.sub.1-6 alkyl, —SH, —SC.sub.1-6 alkyl, —NH.sub.2, —NH—(C.sub.1-6 alkyl), and —N(C.sub.1-6 alkyl).sub.2; R.sup.6 is selected from the group consisting of H, halogen, —OH, —SH, cyano, nitro, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.2-5 alkenyl, —S(O).sub.m—(C.sub.1-6 alkyl), —O—[(C.sub.1-6 alkylene)-O].sub.n—(C.sub.1-6 alkyl), —O—(C.sub.3-6 cycloalkyl), —O-(3-6 membered heterocyclyl), —S(O).sub.m—(C.sub.3-6 cycloalkyl), —S(O).sub.m-(3-10 membered heterocyclyl), —NH.sub.2, —NH—(C.sub.1-6 alkyl), —N(C.sub.1-6 alkyl).sub.2, 3-10 membered heterocyclyl, C.sub.6-14 aryl and 5-14 membered heteroaryl; optionally, the C.sub.1-6 alkyl, cycloalkyl, heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl are each independently substituted by one or more substituents selected from the group consisting of halogen, —OH, —OC.sub.1-6 alkyl, halogenated C.sub.1-6 alkyl, —O-halogenated C.sub.1-6 alkyl, —SH, —SC.sub.1-6 alkyl, —NH.sub.2, —NH—(C.sub.1-6 alkyl), and —N(C.sub.1-6 alkyl).sub.2; m is any integer from 0 to 2, and n is any integer from 0 to 10.

2. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are each independently selected from the group consisting of H, halogen, C.sub.1-6 alkyl, halogenated C.sub.1-6 alkyl, and —O—[(C.sub.1-6 alkylene)-O].sub.n—(C.sub.1-6 alkyl), wherein n is any integer from 0 to 10.

3. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein X is selected from ethylene, vinylene and cyclopropylene, which are each independently and optionally substituted by one or two substituents selected from the group consisting of halogen, C.sub.1-6 alkyl, halogenated C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, (C.sub.1-6 alkyl), 3-10 membered heterocyclyl, C.sub.6-14 aryl and 5-14 membered heteroaryl; Y is selected from a bond and C—R.sup.6, wherein R.sup.6 is selected from the group consisting of H, halogen, —OH, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, —S(O).sub.m—(C.sub.1-6 alkyl), —O—[(C.sub.1-6 alkylene)-O].sub.n—(C.sub.1-6 alkyl), —O—(C.sub.3-6 cycloalkyl), —O-(3-6 membered heterocyclyl), —NH.sub.2, —NH—(C.sub.1-6 alkyl), —N(C.sub.1-6 alkyl).sub.2 and 3-6 membered heterocyclyl, wherein the C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl and 3-6 membered heterocyclyl are optionally substituted by one or more substituents selected from the group consisting of halogen, —OH, —OC.sub.1-3 alkyl, —SH, —SC.sub.1-3 alkyl, —NH.sub.2, —NH—(C.sub.1-3 alkyl) and —N(C.sub.1-3 alkyl).sub.2; m is 0, 1 or 2, and n is 0, 1, 2, 3, 4, or 5; W is selected from N and S.

4. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein R.sup.6 is selected from the group consisting of H, F, Cl, —OH, C.sub.1-3 alkyl, cyclopropyl, —S(O).sub.m(C.sub.1-3 alkyl), —O—[(C.sub.1-2 alkylene)-O].sub.n—(C.sub.1-3 alkyl), —O—(C.sub.5-6 cycloalkyl), —O-(5-6 membered heterocyclyl), —NH.sub.2, —NH—(C.sub.1-6 alkyl), —N(C.sub.1-3 alkyl).sub.2 and 5-6 membered heterocyclyl, wherein the C.sub.1-3 alkyl, C.sub.1-6 alkyl, cyclopropyl, C.sub.5-6 cycloalkyl and 5-6 membered heterocyclyl are optionally substituted by 1 to 3 substituents selected from F, Cl, —OH and —OCH.sub.3; m is 0, 1 or 2, and n is 0, 1 or 2.

5. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein the compound has the structure of Formula (II) or Formula (III): ##STR00043##

6. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein the compound has the structure of Formula (II-1) or Formula (III-1): ##STR00044## wherein, R.sup.7 is selected from the group consisting of H, halogen, C.sub.1-6 alkyl, halogenated C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, —O—(C.sub.1-6 alkyl), 3-10 membered heterocyclyl, C.sub.6-14 aryl and 5-14 membered heteroaryl.

7. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are methyl.

8. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 3, wherein X is vinylene.

9. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein R.sup.5 is selected from H and —O—[(C.sub.1-2 alkylene)-O].sub.n—(C.sub.1-3 alkyl), and n is 0, 1 or 2.

10. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein R.sup.6 is selected from the group consisting of H, F, Cl, —OH, C.sub.1-3 alkyl, —SCH.sub.3, —O—[(C.sub.1-2 alkylene)-O].sub.n—(C.sub.1-3 alkyl), —N(C.sub.1-3 alkyl).sub.2 and 5-6 membered heterocycloalkyl, wherein the C.sub.1-3 alkyl and 5-6 membered heterocycloalkyl are optionally substituted with 1-3 substituents selected from F, Cl, —OH and —OCH.sub.3; n is 0, 1 or 2.

11. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 6, wherein R.sup.7 is selected from H, halogen and C.sub.1-6 alkyl.

12. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 6, wherein R.sup.7 is H or methyl.

13. The compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof according to claim 1, wherein the compound is selected from: ##STR00045## ##STR00046## ##STR00047## ##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052##

14. A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of the compound according to claim 1 or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite, prodrug or mixture thereof, and one or more pharmaceutically acceptable carriers.

15. A pharmaceutical composition, comprising a prophylactically or therapeutically effective amount of the compound according to claim 13 or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite, prodrug or mixture thereof, and one or more pharmaceutically acceptable carriers.

16. A kit product, comprising the compound according to claim 1, or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite, prodrug, or mixture thereof, and an optional package insert.

17. A kit product, comprising the pharmaceutical composition according to claim 14, and an optional package insert.

18. A method for preparing the compound according to claim 1, comprising the following steps: ##STR00053## wherein, V represents halogen or C.sub.1-3 alkyl sulfonic ester group optionally substituted by halogen; Z is selected from H, Cl, Br, I and —P(O)(OEt).sub.2.

19. A method for activating PPAR in a cell, comprising the step of contacting the cell with an effective amount of the compound according to claim 1 or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite, prodrug or mixture thereof.

20. A method for treating a PPAR-related disease or condition, comprising administering to a subject having a disease associated with PPAR an effective amount of the compound according to claim 1 or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite, prodrug or mixture thereof.

21. The method according to claim 20, wherein the PPAR is PPAR α and/or PPAR δ.

22. The method according to claim 20, wherein the disease or condition is a liver disease.

23. The method according to claim 20, wherein the disease or condition is liver fibrosis or fatty liver disease.

24. The method according to claim 20, wherein the disease or condition is NAFLD.

25. The method according to claim 20, wherein the disease or condition is SFL or NASH.

26. A method for treating a PPAR-related disease or condition, comprising administering to a subject having a disease associated with PPAR an effective amount of the compound according to claim 13 or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite, prodrug or mixture thereof.

27. The method according to claim 26, wherein the PPAR is PPAR α and/or PPAR δ.

28. The method according to claim 26, wherein the disease or condition is a liver disease.

29. The method according to claim 26, wherein the disease or condition is liver fibrosis or fatty liver disease.

30. The method according to claim 26, wherein the disease or condition is NAFLD.

31. The method according to claim 26, wherein the disease or condition is SFL or NASH.

Description

EXAMPLES

(1) The invention is further illustrated in detail by the following examples and test examples, which are not intended to limit the scope of the invention, and may be modified without departing from the scope of the invention.

(2) Agilent (ESI) mass spectrometer (manufacturer: Agilent, model: Agilent 6120B) is used for MS determination.

(3) Shimadzu LC-8A liquid chromatograph (YMC, ODS, 250×20 mm column) is used for conducting preparative high-performance liquid chromatograph method.

(4) GF 254 (0.4-0.5 nm) silica gel plate manufactured in Yantai is used for thin layer chromatography purification.

(5) The reaction is monitored by thin layer chromatography (TLC) or LC-MS, and the developing system includes but is not limited to dichloromethane and methanol system, n-hexane and ethyl acetate system, petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, or by adding triethylamine.

(6) For the column chromatography, Qingdao Haiyang 200˜300 mesh silica gel is generally used as stationary phase. The eluent system includes, but is not limited to, dichloromethane and methanol system, and n-hexane and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, or by adding a small amount of triethylamine or the like.

(7) Unless otherwise specified in the examples, the reaction temperature is room temperature (20° C. to 30° C.).

(8) The reagents used in the examples are purchased from Acros Organics, Aldrich Chemical Company or Topbiochem LTD. etc.

(9) Abbreviations as used herein have the following meanings:

(10) TABLE-US-00001 Abbreviations Meaning AcCl Acetyl chloride Ac.sub.2O Acetic anhydride AlCl.sub.3 Aluminium trichloride aq. Aqueous solution Boc Tert-butoxycarbonyl CF.sub.3SO.sub.3H Trifluoromethanesulfonic acid DCM Dichloromethane DIPEA Diisopropylethylamine BBr.sub.3 Boron tribromide DMF N,N-dimethylformamide DMP Dess-Martin periodinane DMSO Dimethyl sulfoxide EA Ethyl acetate Et.sub.3N Triethylamine EtOAc Ethyl acetate EtOH Ethanol K.sub.2CO.sub.3 Potassium carbonate TFA Trifluoroacetic acid H.sub.2 Hydrogen HBr Hydrogen bromide HCl Hydrogen chloride H.sub.2O Water LAH Lithium aluminum hydride CH.sub.3MgBr Methyl magnesium bromide LC-MS Liquid chromatography-mass spectrometry LDA Lithium diisopropylamide MeCN Acetonitrile MeOH Methanol MeONa Sodium methoxide NaBH.sub.4 Sodium borohydride NMP N-methyl pyrrolidone Cs.sub.2CO.sub.3 Cesium carbonate Na.sub.2CO.sub.3 Sodium carbonate NaOH Sodium hydroxide KOH Potassium hydroxide Na.sub.2SO.sub.4 Sodium sulfate m-CPBA meta-chloroperoxybenzoic acid Pd/C Palladium carbon NaSMe Sodium methyl mercaptide Py Pyridine SOCl.sub.2 Thionyl chloride rt Room temperature THF Tetrahydrofuran TLC Thin layer

Example 1: Preparation of tert-butyl 2-(4-formyl-2,6-dimethylphenoxy)-2-methylpropionate (Int1)

(11) ##STR00021##

(12) To SM1 3,5-dimethyl-4-hydroxybenzaldehyde (100 g, 0.67 mol) dissolved in DMF (800 ml) was added cesium carbonate (543 g, 1.67 mmol). The system was heated to 100° C. and reacted for 30 min, followed by dropwise addition of tert-butyl 2-bromoisobutyrate (297 g, 1.33 mmol), then reacted at 120° C. for 8 h. No progression of the reaction was monitored by LC-MS. The reaction solution was poured into ice water for separation. The aqueous phase was extracted with ethyl acetate. The organic phases were combined, dried with anhydrous sodium sulfate, and filtered. The filtrate was concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound Int 1 (31 g), with a yield of 16%.

Example 2: Preparation of (E)-2-(4-(3-(benzo [b]thien-2-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM1)

(13) ##STR00022##

Step 1: Preparation of tert-butyl (E)-2-(4-(3-(benzo [b] thien-2-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionate (1-2)

(14) Compound 1-1 (150 mg, 0.74 mmol) and Int 1 (216 mg, 0.74 mmol) were dissolved in ethanol (20 mL). The mixture was cooled in an ice-water bath for 10 min, followed by dropwise addition of 10% NaOH (0.35 mL), and reacted overnight. No progression of the reaction was monitored by LC-MS. Water and ethyl acetate were added to the mixture for extraction. The organic phases were combined, washed once with saturated saline solution, dried with sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the target product 2-2 (160 mg) with a yield of 45%.

Step 2: Preparation of (E)-2-(4-(3-(benzo [b] thien-2-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM1)

(15) Compound 1-2 (140 mg, 0.29 mmol) was dissolved in DCM (4.5 mL), cooled in an ice-water bath for 10 min. To the mixture was added TFA (1.5 mL) dropwise, and then reaction was conducted for 1 h. LC-MS was applied to monitor the completion of the reaction. The reaction solution was concentrated and purified by column chromatography to obtain the target product TM1 (67 mg) with a yield of 54%.

(16) MS m/z (ESI): 395 [M+H].sup.+

(17) 1HNMR (400 MHz, DMSO-d6) δ: 12.97 (s, 1H), 8.75 (s, 1H), 8.12-8.02 (m, 2H), 7.93 (d, J=16.0 Hz, 1H), 7.68 (d, J=16.0 Hz, 1H), 7.62 (s, 2H), 7.59-7.47 (m, 2H), 2.25 (s, 6H), 1.41 (s, 6H).

Example 3: Preparation of (E)-2-(4-(3-(quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM2)

(18) ##STR00023##

(19) TM2 was synthesized via a method similar to that described in step 1 to step 2 of Example 2 with a yield of 38%, except that 2-1 was used in step 1 of Example 3 instead of 1-1 in step 1 of Example 2.

(20) MS m/z (ESI): 390 [M+H].sup.+

(21) 1HNMR (400 MHz, DMSO-d6) δ: 12.95 (s, 1H), 9.48 (d, J=2.0 Hz, 1H), 9.28 (d, J=2.0 Hz, 1H), 8.22 (d, J=4.0 Hz, 1H), 8.14 (d, J=4.0 Hz, 1H), 8.00 (d, J=16.0 Hz, 1H), 7.98-7.92 (m, 1H), 7.81-7.71 (m, 2H), 7.64 (s, 2H), 2.25 (s, 6H), 1.41 (s, 6H).

Example 4: Preparation of (E)-2-(4-(3-(2-chloroquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM3)

(22) ##STR00024##

Step 1: Preparation of 1-(2-chloroquinolin-3-yl) ethanol (3-2)

(23) CH.sub.3MgBr (1M) (196 mL, 196 mmol) was dissolved in THF (125 mL), and cooled in ice water bath for 10 min. Then, to the mixture was added dropwise a solution of 2-chloro-3-formylquinoline (25 g, 130 mmol) in THF (550 mL). The reaction mixture was reacted for 4 h with stirring. No progression of the reaction was monitored by LC-MS. The reaction solution was poured into saturated aqueous solution of ammonium chloride, extracted with ethyl acetate. The organic phases were combined, washed once with saturated saline solution, dried with sodium sulfate, filtered and concentrated to obtain the target product 3-2 (28 g), which was directly used in the next reaction without purification.

Step 2: Preparation of 2-chloro-3-acetylquinoline (3-3)

(24) Compound 3-2 (28 g, 130 mmol) was dissolved in DCM (560 mL), cooled in an ice-water bath for 15 min, then added with DMP (83 g, 196 mmol) in batches. The mixture was reacted for 2 h. LC-MS was applied to monitor the completion of the reaction. The reaction solution was poured into aqueous solution of sodium sulfite, filtered with diatomite. The filtrate was extracted with DCM. The organic phases were combined, washed once with saturated saline solution, dried with sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the target product 3-3 (18.5 g) with a yield of 69%.

Steps 3 and 4: Two-step synthesis of (E)-2-(4-(3-(2-chloroquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM3)

(25) TM3 was synthesized via a method similar to that described in step 1 to step 2 of Example 2 with a total yield of 26%, except that 3-3 is used in step 3 of Example 4 instead of 1-1 in step 1 of Example 2.

(26) MS m/z (ESI): 424 [M+H].sup.+.

(27) 1HNMR (400 MHz, DMSO-d6) δ: 12.96 (s, 1H), 8.71 (s, 1H), 8.15 (d, J=4.0 Hz, 1H), 8.06 (d, J=4.0 Hz, 1H), 7.98-7.92 (m, 1H), 7.80-7.72 (m, 1H), 7.50 (s, 2H), 7.45 (d, J=16.0 Hz, 1H), 7.27 (d, J=16.0 Hz, 1H), 2.18 (s, 6H), 1.37 (s, 6H).

Example 5: Preparation of (E)-2-(4-(3-(2-methoxyquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM4)

(28) ##STR00025##

Step 1: Preparation of 2-methoxy-3-acetylquinoline (4-2)

(29) To compound 4-1 (300 mg, 1.46 mmol) dissolved in methanol (5 mL) was added sodium methoxide (5M) (1.46 mL, 7.30 mmol). When the completion of the reaction was monitored by LC-MS, the reaction solution was poured into ice water. The mixture was adjusted with 3N HCl aqueous solution to pH 2, then extracted with ethyl acetate. The organic phases were combined, washed once with saturated saline solution, dried with sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the target product 4-2 (270 mg) with a yield of 92%.

Steps 2 and 3: Two-step synthesis of (E)-2-(4-(3-(2-methoxyquinoline-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM4)

(30) TM4 was synthesized via a method similar to that described in step 1 to step 2 of Example 2 with a total yield of 25%, except that 4-2 is used in step 2 of Example 5 instead of 1-1 in step 1 of Example 2.

(31) MS m/z (ESI): 420 [M+H].sup.+

(32) 1HNMR (400 MHz, DMSO-d6) δ: 12.96 (s, 1H), 8.72 (s, 1H), 8.02 (d, J=4.0 Hz, 1H), 7.85 (d, J=4.0 Hz, 1H), 7.81-7.74 (m, 1H), 7.55-7.48 (m, 1H), 7.47 (d, J=16.0 Hz, 1H), 7.46 (s, 2H), 7.34 (d, J=16.0 Hz, 1H), 4.05 (s, 3H), 2.19 (s, 6H), 1.38 (s, 6H).

Example 6: Preparation of (E)-2-(4-(3-(2-(2-methoxyethoxy) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM5)

(33) ##STR00026##

(34) TM5 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 20%, except that sodium 2-methoxyethanolate is used in step 1 of Example 6 instead of sodium methoxide compound in step 1 of Example 5.

(35) MS m/z (ESI): 464 [M+H].sup.+

(36) 1HNMR (400 MHz, DMSO-d6) δ: 12.99 (s, 1H), 8.59 (s, 1H), 8.05 (d, J=4.0 Hz, 1H), 7.83 (d, J=4.0 Hz, 1H), 7.80-7.75 (m, 1H), 7.55-7.48 (m, 3H), 7.45 (s, 2H), 4.68-4.62 (m, 2H), 3.77-3.71 (m, 2H), 3.25 (s, 3H), 2.17 (s, 6H), 1.36 (s, 6H).

Example 7: Preparation of (E)-2-(4-(3-(2-methylquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-m ethylpropionic acid (TM6)

(37) ##STR00027##

Step 1: Preparation of 2-methyl-3-acetylquinoline (6-2)

(38) To compound 6-1 (1.0 g, 8.25 mmol) dissolved in water (20 mL) was added acetylacetone (826 mg, 8.25 mmol) and reacted at reflux for 5 h. When the completion of the reaction was monitored by LC-MS, the reaction solution was poured into ice water, then extracted with ethyl acetate. The organic phases were combined, washed once with saturated aqueous salt solution, dried with sodium sulfate, filtered and concentrated, and purified by column chromatography to obtain the target product 6-2 (780 mg) with a yield of 51%.

Steps 2 and 3: Two-step synthesis of (E)-2-(4-(3-(2-methylquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-m ethylpropionic acid (TM6)

(39) TM6 was synthesized via a method similar to that described in step 1 to step 2 of Example 2 with a yield of 18%, except that 6-2 is used in step 2 of Example 7 instead of 1-1 in step 1 of Example 2.

(40) MS m/z (ESI): 404 [M+H].sup.+.

(41) .sup.1HNMR (400 MHz, DMSO-d6) δ: 12.97 (s, 1H), 8.69 (s, 1H), 8.08 (d, J=4.0 Hz, 1H), 8.00 (d, J=4.0 Hz, 1H), 7.88-7.80 (m, 1H), 7.68-7.60 (m, 1H), 7.51 (s, 2H), 7.49 (d, J=16.0 Hz, 1H), 7.43 (d, J=16.0 Hz, 1H), 2.72 (s, 3H), 2.20 (s, 6H), 1.38 (s, 6H).

Example 8: Preparation of (E)-2-(4-(3-(2-trifluoromethylquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM7)

(42) ##STR00028##

(43) TM7 was synthesized via a method similar to that described in step 1 to step 3 of Example 7 with a yield of 21%, except that trifluoroacetylacetone is used in step 1 of Example 8 instead of acetylacetone compound in step 1 of Example 7.

(44) MS m/z (ESI): 458 [M+H].sup.+

(45) 1HNMR (400 MHz, DMSO-d6) δ: 12.97 (s, 1H), 8.86 (s, 1H), 8.28-8.20 (m, 2H), 8.10-8.00 (m, 1H), 7.95-7.85 (m, 1H), 7.48 (s, 2H), 7.44 (d, J=16.0 Hz, 1H), 7.28 (d, J=16.0 Hz, 1H), 2.17 (s, 6H), 1.37 (s, 6H).

Example 9: Preparation of (E)-2-(4-(3-(2-dimethylaminoquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM9)

(46) ##STR00029##

Step 1: Preparation of 2-dimethylamino-3-acetylquinoline (9-2)

(47) To compound 9-1 (516 mg, 2.5 mmol) dissolved in DMF (5 mL) was added dimethylamine hydrochloride (200 mg, 2.5 mmol) and potassium carbonate (1042 mg, 7.5 mmol), then heated at 60° C. for 4 h. The mixture was added with water and ethyl acetate for extraction. The organic phase was washed twice with saturated brine, dried and concentrated, and separated by column chromatography to obtain the desired product 9-2 (452 mg) with a yield of 84%.

Step 2: Preparation of (E)-2-(4-(3-(2-dimethylaminoquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid tert-butyl ester (9-3)

(48) Compound 9-2 (100 mg, 0.47 mmol) and Int1 (136 mg, 0.47 mmol) were dissolved in 10 mL of anhydrous ethanol, to the mixture was added 10% sodium hydroxide solution (0.13 mL, 0.35 mmol) dropwise in an ice bath and reacted for 4 h. After TLC showed the completion of the reaction, the reaction solution was concentrated. Dichloromethane was added to the residue for dissolution. The solution was filtered. the mother liquor was concentrated, and separated by preparation plate to obtain the target product 9-3 (154 mg) with a yield of 53%.

Step 3: Preparation of (E)-2-(4-(3-(2-dimethylaminoquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM9)

(49) To 9-3 (154 mg, 0.31 mmol) dissolved in 5 mL of dichloromethane was added trifluoroacetic acid (1 mL) and stirred at room temperature for 2 h. The reaction mixture was concentrated, and lyophilized with water to obtain the desired product TM9 (109 mg) with a yield of 80%.

(50) MS m/z (ESI): 433 [M+H].sup.+

(51) 1HNMR (400 MHz, DMSO-d6) δ: 8.58 (s, 1H), 7.96-7.94 (m, 1H), 7.87 (s, 1H), 7.79-7.76 (s, 1H), 7.69 (d, J=16.0 Hz, 1H), 7.51 (s, 2H), 7.45-7.42 (s, 1H), 7.33 (d, J=16.0 Hz, 1H), 3.12 (s, 6H), 2.20 (s, 6H), 1.38 (s, 6H).

Example 10: Preparation of (E)-2-(4-(3-(2-(1-pyrrolidinyl) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM8)

(52) ##STR00030##

(53) Compound TM8 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 44%, except that pyrrolidine is used in step 1 of Example 10 instead of sodium methoxide in step 1 of Example 5.

(54) MS m/z (ESI): 459 [M+H].sup.+

(55) 1HNMR (400 MHz, DMSO-d6) δ: 8.71 (s, 1H), 8.07-8.05 (m, 1H), 8.01 (d, J=8.0 Hz, 1H), 7.88-7.84 (m, 1H), 7.71 (d, J=16.0 Hz, 1H), 7.52 (s, 3H), 7.33 (d, J=16.0 Hz, 1H), 3.54 (s, 4H), 2.20 (s, 6H), 1.99 (s, 4H), 1.39 (s, 6H).

Example 11: Preparation of (E)-2-(4-(3-(2-(morpholin-4-yl) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM10)

(56) ##STR00031##

Step 1: Preparation of 2-(morpholin-4-yl)-3-formylquinoline (10-2)

(57) To the compound 10-1 (1.0 g, 5.2 mmol) dissolved in DMF (15 mL) was added subsequently morpholine (1.4 g, 15.7 mmol) and potassium carbonate (3.6 g, 26.1 mmol). After 4 h at 90° C., the mixture was added with water and ethyl acetate for extraction. The organic phase was washed twice with saturated saline solution, dried and concentrated, and separated by column chromatography to obtain the target product 10-2 (750 mg) with a yield of 59.5%.

Step 2: Preparation of 1-(2-(morpholin-4-yl) quinolin-3-yl)-ethanol (10-3)

(58) CH.sub.3MgBr (1M) (4.7 mL, 4.7 mmol) was dissolved in THF (5 mL), and cooled in an ice-water bath for 10 min. To the above mixture was added dropwise 10-2 (750 mg, 3.1 mmol) dissolved in THF (10 mL). The reaction mixture was stirred for 2 h. No progression of the reaction was monitored by LC-MS, then the reaction mixture was poured into saturated aqueous solution of ammonium chloride, extracted with ethyl acetate. The organic phases were combined, washed once with saturated saline solution, dried with sodium sulfate, filtered and concentrated to obtain the target product 10-2 (800 mg crude), which was directly used in the next reaction without purification.

Steps 3, 4 and 5: Three-step synthesis of (E)-2-(4-(3-(2-(morpholin-4-yl) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM10)

(59) Compound TM10 was synthesized via a method similar to that described in step 2 to step 4 of Example 4 with a yield of 25%, except that 10-3 is used in step 3 of Example 11 instead of 3-2 in step 2 of Example 4.

(60) MS m/z (ESI): 475 [M+H].sup.+

(61) 1HNMR (400 MHz, DMSO-d6) δ: 12.97 (s, 1H), 8.40 (s, 1H), 7.93 (d, J=8.0 Hz, 1H), 7.80-7.68 (m, 2H), 7.62 (d, J=16.0 Hz, 1H), 7.50 (s, 2H), 7.45-7.35 (m, 2H), 3.70-3.50 (m, 4H), 3.40-3.30 (m, 4H), 2.20 (s, 6H), 1.38 (s, 6H).

Example 12: Preparation of (E)-2-(4-(3-(2-(S)-(3-hydroxypyrrolidin-1-yl) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM53)

(62) ##STR00032##

(63) Compound TM53 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 39%, except that (S)-3-hydroxypyrrolidine is used in step 1 of Example 12 instead of sodium methoxide compound in step 1 of Example 5.

(64) MS m/z (ESI): 475 [M+H].sup.+

(65) 1HNMR (400 MHz, DMSO-d6) δ: 8.40 (s, 1H), 7.86 (d, J=8.0 Hz, 2H), 7.67-7.65 (m, 2H), 7.60 (d, J=16.0 Hz, 1H), 7.52 (s, 2H), 7.34-7.30 (m, 2H), 4.33 (s, 1H), 3.62 (s, 2H), 3.47 (s, 2H), 3.12 (s, 1H), 2.20 (s, 6H), 1.98-1.86 (m, 2H), 1.38 (s, 6H).

Example 13: Preparation of (E)-2-(4-(3-(2-(R)-(3-hydroxypyrrolidin-1-yl)quinolin-3-yl)-3-oxo-1-propen-1-yl))-2, 6-dimethylphenoxy)-2-methylpropionic acid (TM54)

(66) ##STR00033##

(67) Compound TM54 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 46%, except that (R)-3-hydroxypyrrolidine is used in step 1 of Example 13 instead of sodium methoxide compound in step 1 of Example 5.

(68) MS m/z (ESI): 475 [M+H].sup.+

(69) 1HNMR (400 MHz, DMSO-d6) δ: 8.44 (s, 1H), 7.87 (d, J=8.0 Hz, 1H), 7.72-7.67 (m, 2H), 7.62 (d, J=16.0 Hz, 1H), 7.52 (s, 2H), 7.34-7.30 (m, 2H), 4.34 (s, 1H), 3.64 (s, 2H), 3.50 (s, 2H), 3.14 (s, 1H), 2.20 (s, 6H), 1.98-1.87 (m, 2H), 1.38 (s, 6H).

Example 14: Preparation of (E)-2-(4-(3-(2-(4-hydroxypiperidin-1-yl) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM25)

(70) ##STR00034##

(71) Compound TM25 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 26%, except that 4-hydroxypiperidine hydrochloride is used in step 1 of Example 14 instead of sodium methoxide compound in step 1 of Example 5.

(72) MS m/z (ESI): 489 [M+H].sup.+

(73) 1HNMR (400 MHz, DMSO-d6) δ: 12.92 (s, 1H), 8.33 (s, 1H), 7.89 (d, J=16.0 Hz, 1H), 7.72-7.64 (m, 2H), 7.61 (d, J=16.0 Hz, 1H), 7.48 (s, 2H), 7.38-7.31 (m, 2H), 4.71 (d, J=2.0 Hz, 1H), 3.75-3.60 (m, 3H), 3.15-3.10 (m, 2H), 2.19 (s, 6H), 1.80-1.70 (m, 2H), 1.50-1.30 (m, 8H).

Example 15: Preparation of (E)-2-(4-(3-(2-hydroxyquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM12)

(74) ##STR00035##

(75) Compound TM4 (15 mg, 0.04 mmol) was dissolved in DCM (3 mL), and cooled in an ice-water bath for 10 min. To the mixture was added boron tribromide dissolved in dichloromethane (1 mL), and reacted for 30 min. After no progression of the reaction was monitored by LC-MS, the reaction solution was concentrated and purified by column chromatography to obtain the target product TM12 (5 mg) with a yield of 35%.

(76) MS m/z (ESI): 406 [M+H].sup.+

(77) 1HNMR (400 MHz, DMSO-d6) δ: 12.13 (s, 1H), 8.46 (s, 1H), 7.87 (d, J=4.0 Hz, 1H), 7.69 (d, J=16.0 Hz, 1H), 7.65-7.60 (m, 1H), 7.69 (d, J=16.0 Hz, 1H), 7.44-7.36 (m, 3H), 7.28-7.22 (m, 1H), 2.11 (s, 6H), 1.37 (s, 6H).

Example 16: Preparation of (E)-2-(4-(3-(2-(R)-(2,3-dihydroxypropylamino) quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM11)

(78) ##STR00036##

(79) Compound TM11 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 5%, except that (R)-3-amino-1,2-propanediol is used in step 1 of Example 16 instead of sodium methoxide compound in step 1 of Example 5.

(80) MS m/z (ESI): 479 [M+H].sup.+

(81) 1HNMR (400 MHz, DMSO-d6) δ: 9.20 (s, 1H), 9.05-8.98 (m, 1H), 8.01 (d, J=16.0 Hz, 1H), 7.89 (d, J=4.0 Hz, 1H), 7.72-7.62 (m, 2H), 7.56 (s, 2H), 7.52 (d, J=4.0 Hz, 1H), 7.26 (t, J=7.2 Hz, 1H), 5.10 (d, J=2.0 Hz, 1H), 4.80 (t, J=4.0 Hz, 1H), 3.85-3.65 (m, 3H), 3.35-3.30 (m, 2H), 2.28 (s, 6H), 1.37 (s, 6H).

Example 17: Preparation of (E)-2-(4-(3-(2-methylthioquinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM13)

(82) ##STR00037##

(83) Compound TM13 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 17%, except that, in step 1 of Example 17, sodium methoxide in step 1 of Example 5 is replaced with sodium methyl mercaptide, and methanol is replaced with NMP.

(84) MS m/z (ESI): 436 [M+H].sup.+

(85) 1HNMR (400 MHz, DMSO-d6) δ: 12.97 (s, 1H), 8.97 (s, 1H), 8.09 (d, J=3.6 Hz, 1H), 7.95 (d, J=8.4 Hz, 1H), 7.90-7.82 (m, 1H), 7.68 (d, J=16.0 Hz, 1H), 7.64-7.52 (m, 4H), 2.58 (s, 3H), 2.22 (s, 6H), 1.39 (s, 6H).

Example 18: Preparation of (E)-2-(4-(3-(2-chloro-7-methoxy-quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM29)

(86) ##STR00038##

(87) Compound TM29 was synthesized via a method similar to that described in step 1 to step 4 of Example 4 with a yield of 7%, except that 29-1 is used in step 1 of Example 18 instead of 3-1 in step 1 of Example 4.

(88) MS m/z (ESI): 454 [M+H].sup.+

(89) .sup.1HNMR (400 MHz, DMSO-d6) δ: 8.64 (s, 1H), 8.07 (d, J=9.2 Hz, 1H), 7.50-7.41 (m, 4H), 7.40-7.36 (m, 1H), 7.28 (d, J=16.0 Hz, 1H), 3.97 (s, 3H), 2.22 (s, 6H), 1.33 (s, 6H).

Example 19: Preparation of (E)-2-(4-(3-(2,7-dimethoxy-quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM28)

(90) ##STR00039##

(91) Compound TM28 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 30%, except that 28-1 is used in step 1 of Example 19 instead of 4-1 in step 1 of Example 5.

(92) MS m/z (ESI): 450 [M+H].sup.+

(93) 1HNMR (400 MHz, DMSO-d6) δ: 12.94 (s, 1H), 8.50 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.55-7.35 (m, 4H), 7.25-7.20 (m, 1H), 7.17-7.10 (m, 1H), 4.06 (s, 3H), 3.94 (s, 3H), 2.22 (s, 6H), 1.38 (s, 6H).

Example 20: Preparation of (E)-2-(4-(3-(2-chloro-6-methoxy-quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethylphenoxy)-2-methylpropionic acid (TM27)

(94) ##STR00040##

(95) Compound TM27 was synthesized via a method similar to that described in step 1 to step 4 of Example 4 with a yield of 22%, except that 27-1 is used in step 1 of Example 20 instead of 3-1 in step 1 of Example 4.

(96) MS m/z (ESI): 454 [M+H].sup.+

(97) 1HNMR (400 MHz, DMSO-d6) δ: 12.96 (s, 1H), 8.55 (s, 1H), 7.97 (d, J=8 Hz, 1H), 7.60-7.52 (m, 2H), 7.50 (s, 2H), 7.44 (d, J=16.0 Hz, 1H), 7.27 (d, J=16.0 Hz, 1H), 3.92 (s, 3H), 2.18 (s, 6H), 1.37 (s, 6H).

Example 21: Preparation of (E)-2-(4-(3-(2,6-dimethoxy-quinolin-3-yl)-3-oxo-1-propen-1-yl)-2,6-dimethyl phenoxy)-2-methylpropionic acid (TM26)

(98) ##STR00041##

(99) Compound TM26 was synthesized via a method similar to that described in step 1 to step 3 of Example 5 with a yield of 32%, except that 26-1 is used in step 1 of Example 21 instead of 4-1 in step 1 of Example 5.

(100) MS m/z (ESI): 450 [M+H].sup.+

(101) .sup.1HNMR (400 MHz, DMSO-d6) δ: 12.95 (s, 1H), 8.42 (s, 1H), 7.78 (d, J=8.0 Hz, 1H), 7.50-7.40 (m, 5H), 7.36 (d, J=16.0 Hz, 1H), 4.01 (s, 3H), 3.87 (s, 3H), 2.19 (s, 6H), 1.38 (s, 6H).

Pharmacological Test

Test Example 1: Experiment of the Compounds on Activation of Transient Transfected PPAR α in HEK293 Cells

(102) Reagent:

(103) Plasmid: pcDNA3. 1(+)-GAL4-hPPAR α, customized by Nanjing Kebai Biotechnology Co., Ltd.

(104) Liposome: PGL4.35, customized by Nanjing Kebai Biotechnology Co., Ltd.

(105) Cell: human embryonic kidney cells HEK293, purchased from ATCC.

(106) Transfection reagent: Lipofectamine Reagent 3000 purchased from Invitrogen.

(107) Assay kit: Bright Glo™ Luciferase Assay System purchased from Promega.

(108) Testing Method:

(109) HEK293 cells were incubated in DMEM medium containing 10% of fetal bovine serum at 37° C. in the presence of 5% of CO.sub.2. 3×10.sup.5/ml cells were plated in each well of the 6-well plate. When the cell convergence degree reached 50%-80%, 5 μg of liposome PGL4.35 and 5 μg of expression plasmid pcDNA3.1(+)-GAL4-hPPAR α were added to transfect the cells for 24 h and then the cells were collected. The transfected cells were plated in the 96-well plate and the compounds of the application in different concentrations were added, incubated for 24 h, and Bright Glo™ Luciferase Assay System reagent was added for luciferase assay. In cells transfected with plasmids and treated with compounds, luciferase activity was increased. The induction of luciferase activity indicated that the compound of the application is PPAR α agonist. EC.sub.50 values of the compound of the application on transfected HEK293 cells were calculated by GraphPad software, and the results were shown in Table 1.

(110) TABLE-US-00002 TABLE 1 Activation of transient transfected PPAR α in HEK293 cells Compound EC.sub.50 (nM) Example 2 1211.46 Example 3 1933.28 Example 4 277.91 Example 5 28.32 Example 6 52.81 Example 7 751.92 Example 8 691.69 Example 9 307.29 Example 10 76.95 Example 11 584.93 Example 17 122.57 Example 18 8.62 Example 19 1.33 Example 20 459.85 Example 21 5.40

(111) As indicated in the data in Table 1, the compound of the application has strong agonist activity on PPAR α: EC.sub.50 of the test compound is less than 2 μM.

Test Example 2: Experiment of the Compounds on Activation of Transient Transfected PPAR δ in HEK293 Cells

(112) Reagent:

(113) Plasmid: pcDNA3. 1(+)-GAL4-hPPAR δ, customized by Nanjing Kebai Biotechnology Co., Ltd.

(114) Liposome: PGL4.35, customized by Nanjing Kebai Biotechnology Co., Ltd.

(115) Cell: human embryonic kidney cells HEK293, purchased from ATCC.

(116) Transfection reagent: Lipofectamine Reagent 3000 purchased from Invitrogen.

(117) Assay kit: Bright Glo™ Luciferase Assay System purchased from Promega.

(118) Testing Method:

(119) HEK293 cells were incubated in DMEM medium containing 10% of fetal bovine serum at 37° C. in the presence of 5% of CO.sub.2. 3×10.sup.5/ml cells were plated in each well of the 6-well plate. When the cell convergence degree reached 50%-80%, 5 jag of liposome PGL4.35 and 5 jag of expression plasmid pcDNA3.1(+)-GAL4-hPPAR δ were added to transfect the cells for 24 h and then the cells were collected. Then, the transfected cells were plated in the 96-well plate and the compounds of the application in different concentrations were added, incubated for 24 h, and Bright Glo™ Luciferase Assay System reagent for luciferase assay was added. In cells transfected with plasmids and treated with compounds, luciferase activity was increased. The induction of luciferase activity indicated that the compound of the application is PPAR δ agonist. EC.sub.50 value of the compound of the application on transfected HEK293 cells were calculated by GraphPad software, and the results were shown in Table 2.

(120) TABLE-US-00003 TABLE 2 Activation of transient transfected PPAR δ in HEK293 cells Compound EC.sub.50 (nM) Example 2 6130.95 Example 4 3441.71 Example 5 555.77 Example 6 373.60 Example 9 6699.54 Example 10 2510.76 Example 11 4426.63 Example 18 809.57 Example 19 795.87 Example 21 674.55

(121) As indicated in the data in Table 2, the compound of the application has strong agonist activity on PPAR δ: EC.sub.50 of the test compound is less than 10 μM.

Test Example 3: Cytotoxicity Test of Compound on HepG2 & Hek293 Cells

(122) Reagent:

(123) Cells: human hepatocyte HepG2, purchased from ATCC; Human embryonic kidney cells HEK293: purchased from ATCC;

(124) Assay reagent: CellTiter Glo® Luminescent Cell Viability Assay, purchased from Promega.

(125) Testing Method:

(126) HepG2 and HEK293 cells were incubated in DMEM/F12 medium containing 10% of fetal bovine serum respectively. A proper amount of cells were plated into a 96-well plate, the plate was placed in an incubator overnight, and the culture medium was removed, and replaced with a complete culture medium containing the compound of the application, and incubated for 3 days. On day 4, an assay reagent CellTiter Glo was added to each well, and the relative luminous unit (RLU) of each well was determined by chemiluminescence. CC.sub.50 values of the compound of the application to HepG2 and HEK293 cells were calculated by GraphPad software, and the results were shown in Table 3.

(127) TABLE-US-00004 TABLE 3 Cytotoxic effect of compound on HepG2 & HEK293 cells HepG2-CC.sub.50 HEK293-CC.sub.50 Compound (μM) (μM) Example 2 ~7.27 ~8.14 Example 4 11.33 13.57 Example 5 12.96 7.13 Example 6 >10 >10 Example 7 >10 >10 Example 8 >10 >10

(128) As indicated from the data in Table 3, CC.sub.50 of toxicity of the compound of the invention to HepG2 cells and HEK293 cells was of μM grade. All test compounds were less cytotoxic to HepG2 cells and HEK293 cells.

Test Example 4: In Vitro Safety Test

(129) The effect of the compound on hERG potassium ion channel was tested by Predictor™ hERG fluorescence polarization. The test results were shown in Table 4:

(130) TABLE-US-00005 TABLE 4 hERG test results Compound IC.sub.50 (μM) Example 2 8.24 Example 3 >10 Example 4 >10 Example 5 >10 Example 6 >10 Example 8 >10 Example 9 >10 Example 10 >10 Example 17 >10 Example 18 >10 Example 19 >10 Example 21 >10

(131) As indicated from the results in Table 4, IC.sub.50 of the compound of the invention for hERG was greater than 8 μM. Therefore, the compound of the invention had no obvious inhibitory effect on hERG, nor potential safety hazard causing prolongation of cardiac QT interval.

(132) In addition to those described herein, various modifications of the invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. All references cited in this application (including all patents, patent applications, journal articles, books and any other publications) are incorporated herein by reference in their entirety.