DIMETHYLSULFOXIMINE DERIVATIVE

Abstract

Disclosed in the present invention is a series of dimethylsulfoximine derivatives, and specifically disclosed are a compound represented by formula (II) and a pharmaceutically acceptable salt thereof.

##STR00001##

Claims

1. A compound represented by formula (II) or a pharmaceutically acceptable salt thereof, ##STR00053## wherein, X is selected from O and NR.sub.b; R.sub.1 and R.sub.4 are each independently selected from H, C.sub.1-3 alkyl, phenyl and 5- to 6-membered heteroaryl, and the C.sub.1-3 alkyl, phenyl and 5- to 6-membered heteroaryl are optionally substituted with 1, 2 or 3 R.sub.a; R.sub.2 and R.sub.3 are each independently selected from H, NH.sub.2, halogen and C.sub.1-3 alkyl; or R.sub.1, R.sub.2 together with the carbon atoms to which they are attached to form C.sub.4-5 cycloalkyl and C.sub.4-5 cycloalkenyl; or R.sub.3, R.sub.4 together with the carbon atoms to which they are attached to form C.sub.4-5 cycloalkyl and C.sub.4-5 cycloalkenyl; R.sub.5 is selected from H, F, Cl, D and CN; R.sub.a is each independently selected from H, C.sub.1-3 alkoxy and CN; R.sub.b is selected from H, CN and C.sub.1-3 alkyl; ring A is selected from 5-membered heteroaryl; the 5- to 6-membered heteroaryl and 5-membered heteroaryl contain 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from —NH—, —O—, —S— and N.

2. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the compound has a structure represented by formula (II-1) or formula (II-2): ##STR00054## wherein, ring A, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are as defined in claim 1.

3. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the compound has a structure represented by formula (I-a) or formula (II-a): ##STR00055## wherein, ring A, R.sub.a and R.sub.5 are as defined in claim 1.

4. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the compound has a structure represented by formula (I-b) or formula (II-b): ##STR00056## wherein, ring A and R.sub.5 are as defined in claim 1.

5. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the compound has a structure represented by formula (I-c) or formula (II-c): ##STR00057## wherein, ring A, R.sub.a and R.sub.5 are as defined in claim 1.

6. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the compound has a structure represented by formula (III): ##STR00058## wherein, T.sub.1 is selected from N and CH; X and R.sub.5 are as defined in claim 1.

7. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.a is selected from H, OCH.sub.3 and CN.

8. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.1 is selected from ##STR00059##

9. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.2 is selected from H.

10. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.3 is selected from H.

11. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.4 is selected from ##STR00060##

12. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.1, R.sub.2 together with the carbon atoms to which they are attached to form ##STR00061##

13. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.3, R.sub.4 together with the carbon atoms to which they are attached to form ##STR00062##

14. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the structural moiety ##STR00063## is selected from ##STR00064##

15. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, ring A is selected from thienyl and thiazolyl.

16. The compound or the pharmaceutically acceptable salt thereof according to claim 15, wherein, ring A is selected from ##STR00065##

17. A compound represented by the following formula or a pharmaceutically acceptable salt thereof, selected from: ##STR00066## ##STR00067##

18. The compound or the pharmaceutically acceptable salt thereof according to claim 17, wherein the compound is selected from, ##STR00068##

19. A method of inhibiting NLRP3 in a subject in need thereof, comprising: administering an effective amount of the compound or the pharmaceutically acceptable salt thereof according to claim 1 to the subject.

20. A method for treating inflammation in a subject in need thereof, comprising: administering an effective amount of the compound or the pharmaceutically acceptable salt thereof according to claim 1 to the subject.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0114] FIG. 1: Results of the inhibition experiment of the inflammatory cytokine IL-6 in APLV.

[0115] FIG. 2: Results of the inhibition experiment of the inflammatory cytokine IL-1β in APLV.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0116] The present disclosure is described in detail by the embodiments below, but it does not mean that there are any adverse restrictions on the present disclosure. The present disclosure has been described in detail herein, and its specific embodiments have also been disclosed; for one skilled in the art, it is obvious to make various modifications and improvements to the embodiments of the present disclosure without departing from the spirit and scope of the present disclosure.

Embodiment 1

[0117] ##STR00042##

[0118] Step 1: Compound 1-1 (3.2 g, 12.2 mmol) and triethylamine (1.9 g, 18.4 mmol) were dissolved in dichloromethane (100 mL), then compound 1-2 (2.66 g, 13.46 mmol) was added, and the reaction was stirred at 25° C. for 2 hours. The reaction mixture was concentrated after the reaction was completed, and then the crude product was separated by column chromatography (petroleum ether:ethyl acetate=3:1) to obtain compound 1-3. MS ESI calculated for C.sub.18H.sub.16BrNO.sub.2S.sub.2 [M+H, M+H+2].sup.+ 422, 424, found 422, 424.

[0119] Step 2: Compound 1-3 (2.8 g, 22.6 mmol) was dissolved in dioxane (30 mL), and then 2-(di-tert-butylphosphino)biphenyl (282.6 mg, 947.1 μmol), compound 1-4 (661.6 mg, 7.1 mmol), sodium tert-butoxide (682.6 mg, 7.1 mmol) and tris(dibenzylideneacetone)dipalladium (433.6 mg, 473.5 μmol) were added, stirred at 80° C. for 1 hour. The reaction mixture was cooled and filtered after the reaction was completed, and the filtrate was concentrated, and then the crude product was purified by column chromatography (petroleum ether:ethyl acetate=3:1) to obtain compound 1-5. MS ESI calculated for C.sub.20H.sub.22N.sub.2O.sub.3S.sub.3 [M+H].sup.+ 435, found 435.

[0120] Step 3: Compound 1-5 (1.0 g, 2.3 mmol) was dissolved in dichloromethane (10 mL), and sulfuric acid (concentration of 98%, 3.5 mL) was added dropwise at 0° C., and the reaction mixture was stirred at 25° C. for 1 hour, and then poured into ice water (50 mL), then extracted with dichloromethane (150 mL). The organic phase was concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain compound 1-6. MS ESI calculated for C.sub.6H.sub.10N.sub.2O.sub.3S.sub.3 [M+H].sup.+ 255, found 255.

[0121] Step 4: Sodium hydride (30.3 mg, 758.3 μmol, purity of 60%) was added to a solution of compound 1-6 (150.0 mg, 589.7 μmol) in tetrahydrofuran (10 mL), stirred at 0° C. for 10 min, and then a solution of compound 1-7 (123.3 mg, 619.2 μmol) in tetrahydrofuran (10 mL) was added to the system and stirred at 25° C. for 0.5 hours. The reaction was quenched with dilute hydrochloric acid (1 mol/L, 1 mL) after the reaction was completed, then extracted with ethyl acetate (50 mL*2). The organic phase was concentrated, and the crude product was purified by column chromatography (ethyl acetate:ethanol=10:1) to obtain compound 1. MS ESI calculated for C.sub.19H.sub.23N.sub.3O.sub.4S.sub.3 [M+H].sup.+ 454, found 454. .sup.1H NMR (400 MHz, CD.sub.3CN) δ ppm 8.42 (brs, 1H), 8.19 (s, 1H), 8.09 (s, 1H), 7.01 (d, J=4.0 Hz, 1H), 6.38 (d, J=4.0 Hz, 1H), 3.23 (s, 6H), 2.87 (brt, J=7.4 Hz, 4H), 2.66 (brt, J=7.4 Hz, 4H), 2.00-2.07 (m, 4H).

Embodiment 2

[0122] ##STR00043##

[0123] Step 1: Compound 2-1 (500.0 mg, 2.3 mmol) and compound 2-2 (357.0 mg, 2.3 mmol) were dissolved in dioxane (40 mL)/water (8 mL), and then [1,1′-bis(diphenylphosphino)ferrocene] dichloropalladium(II) dichloromethane complex (191.1 mg, 234.0 μmol) and potassium carbonate (646.8 mg, 4.7 mmol) were added, and the reaction was stirred at 100° C. for 2 hours, then cooled to room temperature, extracted with water (50 mL) and ethyl acetate (150 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1) to obtain compound 2-3. MS ESI calculated for C.sub.15H.sub.18N.sub.2O [M+H].sup.+ 243, found 243.

[0124] Step 2: Compound 2-3 (95.0 mg, 392.1 mmol) was dissolved in tetrahydrofuran (10 mL), and then triphosgene (50.0 mg, 168.6 μmol) and triethylamine (119.0 mg, 1.2 mmol) were added at 25° C., and the reaction was stirred at 25° C. for 0.5 hours. After the reaction was completed, the reaction mixture was filtered to obtain a solution of compound 2-4 in tetrahydrofuran, which was directly used in the next step. MS ESI calculated for C.sub.16H.sub.16N.sub.2O.sub.2 [M+H].sup.+ 269, found 269.

[0125] Step 3: Sodium hydride (39.3 mg, 982.9 μmol, purity of 60%) was added to a solution of compound 1-6 (100.0 mg, 393.2 μmol) in tetrahydrofuran (10 mL), stirred at 0° C. for 10 min, and then a solution of compound 2-4 (105.49 mg, 393.16 μmol) in tetrahydrofuran (10 mL) was added to the system and stirred at 25° C. for 0.5 hours. The reaction was quenched with dilute hydrochloric acid (1 mol/L, 1 mL) after the reaction was completed, then extracted with ethyl acetate (50 mL*2). The organic phase was concentrated, and the crude product was separated by preparative high performance liquid chromatography (chromatographic column: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile %: 15%-40%, 9.5 min) to obtain compound 2. MS ESI calculated for C.sub.22H.sub.26N.sub.4O.sub.5S.sub.3 [M+H].sup.+ 523, found 523. .sup.1H NMR (400 MHz, CD.sub.3OD) δ ppm 8.04 (d, J=5.3 Hz, 1H), 7.41-7.43 (m, 2H), 7.17-7.19 (m, 1H), 6.85-6.87 (m, 2H), 6.75 (s, 1H), 6.40 (d, J=4.3 Hz, 1H), 3.94 (s, 3H), 3.13 (s, 6H), 3.04-3.13 (m, 1H), 1.21 (d, J=6.8 Hz, 6H).

Embodiment 3

[0126] ##STR00044## ##STR00045##

[0127] Step 1: Compound 3-1 (9.0 g, 67.1 mmol) was dissolved in dichloromethane (50 mL), and then trifluoromethanesulfonic anhydride (37.8 g, 134.1 mmol) and pyridine (15.9 g, 201.23 mmol) were slowly added at 0° C., and the reaction was stirred at 25° C. for 1 hour, then quenched with water (50 mL) and extracted with dichloromethane (100 mL). The organic phase was concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=20:1) to obtain compound 3-2.

[0128] Step 2: Compound 3-2 (10.0 g, 37.5 mmol) and tert-butyl carbamate (8.8 g, 75.1 mmol) were dissolved in dioxane (150 mL), and then cesium carbonate (24.48 g, 75.12 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (4.3 g, 7.5 mmol) and tris(dibenzylideneacetone)dipalladium (3.4 g, 3.7 mmol) were added under the protection of nitrogen, and the reaction was stirred at 80° C. for 1 hour. The reaction was quenched with water (50 mL) after the reaction was completed, extracted with ethyl acetate (150 mL). The organic phase was concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=5:1) to obtain compound 3-3. MS ESI calculated for C.sub.14H.sub.19NO.sub.2 [M+H].sup.+ 234, found 234.

[0129] Step 3: Compound 3-3 (6.0 g, 25.7 mmol) was dissolved in dichloromethane (50 mL), and trifluoroacetic acid (17.6 g, 154.3 mmol) was added dropwise at 25° C., and the reaction was stirred at 25° C. for 1 hour, then quenched with saturated sodium bicarbonate (200 mL), extracted with dichloromethane (200 mL). The organic phase was concentrated to obtain compound 3-4, which was directly used in the next step. MS ESI calculated for C.sub.9H.sub.11N [M+H].sup.+ 134, found 134.

[0130] Step 4: Compound 3-4 (2.6 g, 19.5 mmol) and triethylamine (2.6 g, 25.4 mmol) were dissolved in dichloromethane (30 mL), then acetic anhydride (2.3 g, 22.5 mmol) was added dropwise, and the reaction was stirred at 25° C. for 16 hours. The reaction was quenched with water (50 mL) after the reaction was completed, then extracted with dichloromethane (150 mL). The organic phase was concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=2:1) to obtain compound 3-5. MS ESI calculated for C.sub.11H.sub.13NO [M+H].sup.+ 176, found 176.

[0131] Step 5: Compound 3-5 (2.9 g, 16.5 mmol) was dissolved in tetrahydrofuran (50 mL), and then p-toluenesulfonic acid (1.6 g, 9.1 mmol) and palladium acetate (185.7 mg, 827.5 μmol) were added, and N-bromosuccinimide (3.2 g, 18.2 mmol) was added after the reaction was stirred at 20° C. for 0.5 hours, and the reaction was continued to stir at 20° C. for 2 hours. The reaction was quenched with water (50 mL) after the reaction was completed, extracted with ethyl acetate (150 mL). After the organic phase was dried over anhydrous sodium sulfate, the organic phase was concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=2:1) to obtain compound 3-6. MS ESI calculated for C.sub.11H.sub.12BrNO [M, M+2].sup.+ 254, 256, found 254, 256.

[0132] Step 6: Compound 3-6 (2.3 g, 9.1 mmol) was dissolved in ethanol (20 mL) and concentrated hydrochloric acid (7 mL, concentration of 37%), and the reaction was stirred at 80° C. for 12 hours. The reaction was quenched with saturated sodium bicarbonate (200 mL) after the reaction was completed, extracted with ethyl acetate (200 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was separated by column chromatography (petroleum ether:ethyl acetate=2:1) to obtain compound 3-7. MS ESI calculated for C.sub.9H.sub.10BrN [M, M+2].sup.+ 212, 214, found 212, 214.

[0133] Step 7: Compound 3-7 (200.0 mg, 943.0 μmol) and compound 2-2 (158.6 mg, 1.0 mmol) were dissolved in dioxane (16 mL)/water (4 mL), and then potassium carbonate (325.8 mg, 2.3 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (69.0 mg, 94.3 μmol) were added, and the reaction was stirred at 80° C. for 2 hours under the protection of nitrogen and then concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain compound 3-8. MS ESI calculated for C.sub.15H.sub.16N.sub.2O [M+H].sup.+ 241, found 241.

[0134] Step 8: Compound 3-8 (103.7 mg, 431.8 μmol) was dissolved in tetrahydrofuran (10 mL), and then triphosgene (55.1 mg, 185.6 μmol) and triethylamine (131.2 mg, 1.3 mmol) were added at 25° C., and the reaction was stirred at 25° C. for 0.5 hours. A solution of compound 3-9 in tetrahydrofuran was obtained by filtration after the reaction was completed, which was directly used in the next step. MS ESI calculated for C.sub.16H.sub.14N.sub.2O.sub.2 [M+H].sup.+ 267, found 267.

[0135] Step 9: Sodium hydride (39.3 mg, 982.9 mol, purity of 60%) was added to a solution of compound 1-6 (100.0 mg, 393.2 μmol) in tetrahydrofuran (10 mL), stirred at 0° C. for 10 min, and then a solution of compound 3-9 (104.7 mg, 393.2 μmol) in tetrahydrofuran (10 mL) was added to the system and stirred at 25° C. for 0.5 hours. The reaction was quenched with dilute hydrochloric acid (1 mol/L, 1 mL) after the reaction was completed, then extracted with ethyl acetate (50 mL×2). The organic phase was concentrated, and the crude product was separated by preparative high performance liquid chromatography (chromatographic column: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile %: 15%-40%, 9.5 min) to obtain compound 3. MS ESI calculated for C.sub.22H.sub.24N.sub.4O.sub.5S.sub.3 [M+H].sup.+ 521, found 521. .sup.1H NMR (400 MHz, CD.sub.3OD) δ ppm 8.07 (d, J=5.3 Hz, 1H), 7.40-7.44 (m, 2H), 7.12-7.22 (m, 1H), 6.84 (d, J=4.3 Hz, 1H), 6.76 (s, 1H), 6.40 (d, J=4.3 Hz, 1H), 3.93 (s, 3H), 3.13 (s, 6H), 3.00 (t, J=7.4 Hz, 2H), 2.80-2.83 (m, 2H), 2.12 (t, J=7.4 Hz, 2H).

Embodiment 4

[0136] ##STR00046## ##STR00047##

[0137] Step 1: Dibenzylamine (4.0 g, 20.1 mmol) and triethylamine (2.3 g, 22.9 mmol) were added slowly to a solution of compound 4-1 (5.0 g, 19.1 mmol) in dichloromethane (25 mL) in turn at 0° C., and the reaction was stirred at 25° C. for 12 hours. The reaction mixture was added to water (25 mL), extracted with dichloromethane (50 mL*3), and the organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and purified by column chromatography (petroleum ether: ethyl acetate=20:1-1:1) to obtain compound 4-2, which was directly used in the next step. MS ESI calculated for C.sub.18H.sub.16BrNO.sub.2S.sub.2 [M+H; M+H+2].sup.+ 422; 424, found 422; 424.

[0138] Step 2: Compound 4-2 (3.4 g, 8.1 mmol) was dissolved in 1,4-dioxane (40 mL), and then compound 1-4 (824.8 mg, 8.9 mmol), tris(dibenzylideneacetone)dipalladium (737.2 mg, 805.0 mol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (931.5 mg, 1.6 mmol) and cesium carbonate (5.3 g, 16.1 mmol) were added in turn, and the system was replaced with nitrogen for three times. The reaction was stirred at 110° C. for 12 hours and then cooled to 25° C. and filtered, and the filtrate was concentrated and purified by column chromatography (petroleum ether:ethyl acetate=10:1-0:1) to obtain compound 4-3, which was directly used in the next step. MS ESI calculated for C.sub.20H.sub.22N.sub.2O.sub.3S.sub.3 [M+H].sup.+ 435, found 435.

[0139] Step 3: Concentrated sulfuric acid (5.4 g, 54.0 mmol, concentration of 98%) was added to a solution of compound 4-3 (2.4 g, 5.4 mmol) in dichloromethane (20 mL) at 0° C., and the reaction was stirred at 20° C. for 2 hours. About 30 g of ice was added to the reaction mixture, and the pH was adjusted to 5-6 with sodium hydroxide solid and then the mixture was extracted with a mixed solution of dichloromethane:methanol=10:1 (30 mL*3). The organic phase was dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (dichloromethane:methanol=30:1-10:1) to obtain compound 4-4, which was directly used in the next step. MS ESI calculated for C.sub.6H.sub.10N.sub.2O.sub.3S.sub.3 [M+H].sup.+ 255, found 255.

[0140] Step 4: A solution of compound 4-4 (1.0 g, 3.9 mmol) in tetrahydrofuran (20 mL) was cooled to 0° C., and sodium hydride (346.0 mg, 8.7 mmol, purity of 60%) was added, and then stirred for 0.5 hours. tert-Butyldimethylsilyl chloride (711.1 mg, 4.7 mmol) was added and warmed up to 25° C. and stirred for 12 hours. The reaction was cooled to 0° C., quenched with saturated aqueous ammonium chloride solution (10 mL), extracted with ethyl acetate (20 mL*3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, purified by column chromatography (petroleum ether:ethyl acetate=3:1-0:1) to compound 4-5, which was directly used in the next step. MS ESI calculated for C.sub.12H.sub.24N.sub.2O.sub.3S.sub.3Si [M+H].sup.+ 369, found 369.

[0141] Step 5: Dichlorotriphenylphosphorane (2.0 g, 6.1 mmol) was dissolved in chloroform (20 mL), and triethylamine (1.1 g, 10.8 mmol) was added at 0° C., stirred for 15 min, then compound 4-5 (1.0 g, 2.7 mmol) was added, stirred at 0° C. for 0.5 hours, and then added to a saturated ammonia tetrahydrofuran solution (20 mL) which was precooled to −40° C., naturally warmed up to 25° C. and stirred for 12 hours. After the reaction was completed, the mixture was concentrated directly, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Welch Xtimate C18 250*70 mm: 10 μm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile %: 30%-57%, 30 min) to obtain compound 4-6, which was directly used in the next step. MS ESI calculated for C.sub.12H.sub.25N.sub.3O.sub.2S.sub.3Si [M+H].sup.+ 368, found 368.

[0142] Step 6: Compound 4-6 (150.0 mg, 408.0 μmol) was dissolved in tetrahydrofuran (10 mL), and sodium hydride (32.6 mg, 816.1 mol, purity of 60%) was added at 0° C. and stirred for 0.5 hours, then a solution of compound 1-7 (81.3 mg, 408.0 μmol) in tetrahydrofuran (10 mL) was added and the reaction was continued to stir at 25° C. for 1 hour. The reaction mixture was cooled to 0° C. and quenched with water (2 mL) to obtain a solution of compound 4-7, which was directly used in the next step. MS ESI calculated for C.sub.25H.sub.38N.sub.4O.sub.3S.sub.3Si [M+H].sup.+ 567, found 567.

[0143] Step 7: Dilute hydrochloric acid (1 mol/L, 10 mL) was added to the solution of compound 4-7 and stirred at 25° C. for 0.5 hours. The mixture was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Welch Xtimate C18 250*70 mm #10 μm; mobile phase: [water (10 mM ammonium bicarbonate)-acetonitrile]; acetonitrile %: 27%-47%, 25 min) to obtain compound 4. MS ESI calculated for C.sub.19H.sub.24N.sub.4O.sub.3S.sub.3[M+H].sup.+ 453, found 453.

[0144] Step 8: Compound 4 (100 mg) was separated by preparative supercritical fluid chromatography (chromatographic column: Chiralcel OD-3 100 mm*4.6 mm I.D., 3 μm); mobile phase: [A: carbon dioxide, B: methanol (0.05% diethylamine)], gradient: B: 5%-40%, 4 min; B %: 40%, 2.5 min; B %: 5%, 1.5 min) to obtain compound 4a (retention time was 4.68 min) and compound 4b (retention time was 5.24 min).

[0145] Compound 4a, .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.26 (s, 1H), 7.47 (s, 2H), 7.29 (d, J=4.0 Hz, 1H), 6.86 (s, 1H), 6.28 (d, J=4.0 Hz, 1H), 3.29 (s, 6H), 2.78 (t, J=7.4 Hz, 4H), 2.69 (t, J=7.4 Hz, 4H), 1.93 (t, J=7.4 Hz, 4H). MS ESI calculated for C.sub.19H.sub.24N.sub.4O.sub.3S.sub.3 [M+H].sup.+ 453, found 453.

[0146] Compound 4b, .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.30 (s, 1H), 7.49 (s, 2H), 7.30 (d, J=4.0 Hz, 1H), 6.88 (s, 1H), 6.30 (d, J=4.0 Hz, 1H), 3.32 (s, 6H), 2.79 (t, J=7.4 Hz, 4H), 2.71 (t, J=7.4 Hz, 4H), 1.94 (t, J=7.4 Hz, 4H). MS ESI calculated for C.sub.19H.sub.24N.sub.4O.sub.3S.sub.3 [M+H].sup.+ 453, found 453.

Embodiment 5

[0147] ##STR00048##

[0148] Step 1: Sodium hydride (43.5 mg, 1.1 mmol, purity of 60%) was added to a solution of compound 4-6 (100.0 mg, 272.0 μmol) in tetrahydrofuran (10.0 mL), stirred at 25° C. for 0.5 hours, and then compound 3-9 (72.4 mg, 272.0 μmol) was added to the system and continued to stir for 1 hour to obtain a reaction mixture of compound 5-1, which was directly used in the next step. MS ESI calculated for C.sub.28H.sub.39N.sub.5O.sub.4S.sub.3Si [M+H].sup.+ 634, found 634.

[0149] Step 2: Concentrated hydrochloric acid (5.0 mL, concentration of 37%) was added dropwise to the reaction mixture of compound 5-1 at 0° C. and stirred for 10 min, extracted with ethyl acetate (30 mL) after the reaction was completed. The organic phase was dried over anhydrous sodium sulfate. The obtained crude product was separated by column chromatography (dichloromethane:methanol=10:1) to obtain compound 5. MS ESI calculated for C.sub.22H.sub.25N.sub.5O.sub.4S.sub.3 [M+H].sup.+ 520, found 520.

[0150] Step 3: Compound 5 (90 mg) was separated by preparative chromatography (chromatographic column: Cellulose-2 100 mm*4.6 mm I.D., 3 μm; mobile phase A: carbon dioxide; B: [0.05% diethylamine-methanol]; gradient: B %: 50%-50%, 25 min) to obtain compound 5a (retention time was 2.58 min) and compound 5b (retention time was 3.82 min).

[0151] Compound 5a, .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ ppm 8.26 (brs, 1H), 8.12 (d, J=5.3 Hz, 1H), 7.42 (brs, 2H), 7.15-7.25 (m, 1H), 7.06-7.14 (m, 2H), 6.95 (brd, J=4.8 Hz, 1H), 6.76 (s, 1H), 6.25 (d, J=4.0 Hz, 1H), 3.88 (s, 3H), 3.32 (s, 6H), 2.92 (t, J=7.4 Hz, 2H), 2.79 (brs, 2H), 1.96-2.03 (m, 2H). MS ESI calculated for C.sub.22H.sub.25N.sub.5O.sub.4S.sub.3 [M+H].sup.+ 520, found 520.

[0152] Compound 5b, .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ ppm 8.26 (brs, 1H), 8.12 (d, J=5.3 Hz, 1H), 7.42 (brs, 2H), 7.15-7.21 (m, 1H), 7.06-7.16 (m, 2H), 6.95 (brd, J=4.8 Hz, 1H), 6.76 (s, 1H), 6.25 (d, J=4.0 Hz, 1H), 3.88 (s, 3H), 3.32 (s, 6H), 2.92 (brt, J=7.4 Hz, 2H), 2.79 (brs, 2H), 1.98-2.03 (m, 2H). MS ESI calculated for C.sub.22H.sub.25N.sub.5O.sub.4S.sub.3 [M+H].sup.+ 520, found 520.

Embodiment 6

[0153] ##STR00049##

[0154] Step 1: Potassium carbonate (5.1 g, 37.0 mmol) was added to a solution of benzyl mercaptan (1.5 g, 12.3 mmol) in N,N-dimethylformamide (30 mL), stirred at 25° C. for 5 min, and then compound 6-1 (3.0 g, 12.4 mmol) was added. The reaction was warmed up to 100° C. and continued to stir for 5 hours, then cooled to 25° C., quenched with water (60 mL), extracted with ethyl acetate (60 mL*3). The combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 6-2, which was directly used in the next step. .sup.1H NMR (400 MHz, CDCl.sub.3): 7.51 (s, 1H), 7.20-7.33 (m, 5H), 4.35 (s, 2H). MS ESI calculated for C.sub.10H.sub.8BrNS.sub.2 [M+H; M+H+2].sup.+ 286; 288, found 286; 288.

[0155] Step 2: Compound 6-2 (1.0 g, 3.5 mmol), acetic acid (10 mL), water (5 mL) and 1,3-dichloro-5,5-dimethylhydantoin (2.8 g, 14.0 mmol) were added to a pre-dried reaction flask and the reaction was stirred at 40° C. for 1.5 hours. The reaction mixture was quenched with water (20 mL) after the reaction was completed, extracted with dichloromethane (20 mL*3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Petroleum ether (1 mL) and ethyl acetate (1 mL) were added to the residue, stirred for 10 minutes and then filtered, and the filtrate was concentrated under reduced pressure to obtain compound 6-3, which was immediately used in the next step.

[0156] Step 3: Compound 6-3 (800.0 mg, 3.1 mmol) was dissolved in 1,2-dichloroethane (10 mL), and dibenzylamine (2.4 g, 12.2 mmol) was added. The reaction was stirred at 80° C. for 12 hours and then cooled to 25° C., quenched by adding water (40 mL) to the reaction mixture, extracted with ethyl acetate (40 mL*3), and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol=20:1) to obtain compound 6-4. MS ESI calculated for C.sub.17H.sub.15BrN.sub.2O.sub.2S.sub.2 [M+H; M+H+2].sup.+ 423; 425, found 423; 425.

[0157] Step 4: Compound 6-4 (390.0 mg, 992.1 mol), 1,4-dioxane (10 mL), compound 1-4 (138.6 mg, 1.5 mmol) and cesium carbonate (969.7 mg, 2.9 mmol) were added to a pre-dried reaction flask, and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (114.8 mg, 198.4 μmol) and tris(dibenzylideneacetone)dipalladium (90.8 mg, 99.2 μmol) were added finally. The reaction was stirred at 110° C. for 2 hours under the protection of nitrogen and then cooled to 25° C., quenched by adding water (20 mL) to the reaction mixture, extracted with ethyl acetate (20 mL*3), and the combined organic phases were washed with saturated brine (20 mL*3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol=10:1) to obtain compound 6-5. MS ESI calculated for C.sub.19H.sub.21N.sub.3O.sub.3S.sub.3 [M+H].sup.+ 436, found 436.

[0158] Step 5: Compound 6-5 (150 mg, 344.3 μmol) was dissolved in dichloromethane (1 mL), and concentrated sulfuric acid (1 mL, concentration of 98%) was added. The reaction was stirred at 25° C. for 0.5 hours. The reaction mixture was slowly poured into ice water (5 mL) after the reaction was completed, and the pH was adjusted to 4-5 with 2 mol/L sodium hydroxide solution, and concentrated under reduced pressure to obtain a residue, and the residue was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound 6-6. MS ESI calculated for C.sub.5H.sub.9N.sub.3O.sub.3S.sub.3 [M+H].sup.+ 256, found 256.

[0159] Step 6: Compound 6-6 (220.0 mg, 86.1 μmol) was dissolved in tetrahydrofuran (1 mL), and sodium hydride (10.3 mg, 258.4 mol, purity of 60%) was added at 0° C. and stirred for 0.5 hours, then compound 1-7 (20.6 mg, 103.3 μmol) was added. The reaction was warmed up to 25° C. and continued to stir for 0.5 hours, quenched with water (0.5 mL) and concentrated under reduced pressure, and the residue was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to obtain compound 6. .sup.1H NMR (400 MHz, CD.sub.3OD) δ ppm 7.10 (brs, 1H), 6.89 (s, 1H), 3.27 (s, 6H), 2.82 (t, J=7.1 Hz, 4H), 2.69-2.76 (m, 4H), 1.96-2.03 (m, 4H); MS ESI calculated for C.sub.18H.sub.22N.sub.4O.sub.4S.sub.3 [M+H].sup.+ 455, found 455.

Embodiment 7

[0160] ##STR00050##

[0161] Step 1: Compound 6-6 (200.0 mg, 783.3 μmol) was dissolved in tetrahydrofuran (20 mL), and sodium hydride (78.3 mg, 1.9 mmol, purity of 60%) was added at 0° C. and stirred for 0.5 hours, then tert-butyldimethylsilyl chloride (141.6 mg, 939.9 μmol) was added and stirred at 25° C. for 1 hour, quenched with saturated ammonium chloride solution (5 mL) after the reaction was completed, and extracted with ethyl acetate (30 mL*2). The combined organic phases were dried over anhydrous sodium sulfate, concentrated and the residue was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound 7-1. MS ESI calculated for C.sub.11H.sub.23N.sub.3O.sub.3S.sub.3Si [M+H].sup.+ 370, found 370.

[0162] Step 2: Triethylamine (260.7 mg, 2.5 mmol) was added dropwise to a solution of dichlorotriphenylphosphorane (429.3 mg, 1.3 mmol) in chloroform (10 mL) at 25° C., stirred for 10 min and then cooled to 0° C. A solution of compound 7-1 (190.0 mg, 515.5 μmol) in chloroform (3 mL) was added and the reaction was continued to stir at 0° C. for 0.5 hours. Ammonia was introduced into the system for 15 min, then the reaction was warmed up to 25° C. and stirred for 1 hour. After the reaction was completed, the mixture was concentrated to obtain compound 7-2, which was directly used in the next step. MS ESI calculated for C.sub.11H.sub.24N.sub.4O.sub.2S.sub.3Si [M+H].sup.+ 369, found 369.

[0163] Step 3: Sodium hydride (78.13 mg, 1.95 mmol, purity of 60%) was added to a solution of compound 7-2 (180.0 mg, 488.32 μmol) in tetrahydrofuran (20 mL), stirred at 25° C. for 0.5 hours, and then compound 1-7 (97.3 mg, 488.3 μmol) was added to the system and continued to stir for 1 hour. After the reaction was completed, the reaction mixture of compound 7-3 was directly used in the next step. MS ESI calculated for C.sub.24H.sub.37N.sub.5O.sub.3S.sub.3Si [M+H].sup.+ 568, found 568.

[0164] Step 4: Concentrated hydrochloric acid (5.0 mL, concentration of 37%) was added dropwise to the reaction mixture of compound 7-3 at 0° C. and stirred for 10 min, extracted with ethyl acetate (30 mL) after the reaction was completed, and the organic phase was dried over anhydrous sodium sulfate. The obtained crude product was separated by column chromatography (dichloromethane:methanol=10:1) to obtain compound 7. MS ESI calculated for C.sub.18H.sub.23N.sub.5O.sub.3S.sub.3 [M+H].sup.+ 454, found 454.

[0165] Step 5: Compound 7 (20 mg) was separated by preparative supercritical fluid chromatography (chromatographic column: Chiralpak AS-3 150 mm*4.6 mm I.D., 3 μm); mobile phase: [A: carbon dioxide, B: ethanol (0.05% diethylamine)]; gradient: B %: 5%-40%, 5 min; B %: 40%, 2.5 min; B %: 5%, 2.5 min) to obtain compound 7a (retention time was 5.53 min) and 7b (retention time was 6.15 min).

[0166] Compound 7a: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ ppm 8.41 (brs, 1H), 7.73 (brs, 2H), 7.23 (s, 1H), 6.87 (s, 1H), 3.35 (s, 6H), 2.78 (brt, J=7.3 Hz, 4H), 2.67 (brs, 4H), 1.86-1.97 (m, 4H). MS ESI calculated for C.sub.18H.sub.23N.sub.5O.sub.3S.sub.3 [M+H].sup.+ 454, found 454.

[0167] Compound 7b: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ ppm 8.42 (brs, 1H), 7.73 (brs, 2H), 7.23 (s, 1H), 6.87 (s, 1H), 3.35 (brs, 6H), 2.78 (brt, J=7.0 Hz, 4H), 2.68 (brs, 4H), 1.93 (brt, J=7.3 Hz, 4H). MS ESI calculated for C.sub.18H.sub.23N.sub.5O.sub.3S.sub.3 [M+H].sup.+ 454, found 454.

Embodiment 8

[0168] ##STR00051## ##STR00052##

[0169] Step 1: Compound 2-1 (2.0 g, 9.3 mmol) and compound 8-1 (2.2 g, 9.3 mmol) were dissolved in dioxane (40 mL)/water (8 mL), and then [1,1′-bis(diphenylphosphino)ferrocene] dichloropalladium(II) dichloromethane complex (762.8 mg, 934.1 μmol) and potassium carbonate (2.6 g, 18.6 mmol) were added, and the reaction was stirred at 100° C. for 2 hours then cooled to 25° C., extracted with water (50 mL) and ethyl acetate (150 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate=2:1) to obtain compound 8-2. MS ESI calculated for C.sub.15H.sub.15N.sub.3 [M+H].sup.+ 238, found 238.

[0170] Step 2: Compound 8-2 (1.0 g, 4.2 mmol) was dissolved in tetrahydrofuran (60 mL), and then triphosgene (537.7 mg, 1.8 mmol) and triethylamine (1.3 g, 12.6 mmol) were added at 25° C., and the reaction was stirred at 25° C. for 0.5 hours. After the reaction was completed, the reaction mixture was filtered to obtain a reaction mixture of compound 8-3 in tetrahydrofuran, which was directly used in the next step. MS ESI calculated for C.sub.16H.sub.13N.sub.3O [M+H].sup.+ 264, found 264.

[0171] Step 3: Sodium hydride (43.5 mg, 1.1 mmol, purity of 60%) was added to the reaction mixture of compound 4-6 (100.0 mg, 272.0 μmol) in tetrahydrofuran (10.0 mL) at 25° C. and stirred for 0.5 hours, then compound 8-3 (71.1 mg, 272.0 μmol) was added to the system and continued to stir for 1 hour. After the reaction was completed, the reaction mixture of compound 8-4 was directly used in the next step. MS ESI calculated for C.sub.28H.sub.36N.sub.6O.sub.3S.sub.3Si [M+H].sup.+ 629, found 629.

[0172] Step 4: Concentrated hydrochloric acid (5 mL, concentration of 37%) was added dropwise to the reaction mixture of compound 8-4 at 0° C. and stirred for 10 min, extracted with ethyl acetate (30 mL) after the reaction was completed. The organic phase was dried over anhydrous sodium sulfate, filtered, and the crude product was concentrated and separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound 8. .sup.1H NMR (400 MHz, CD.sub.3OD) δ ppm 8.67 (d, J=5.5 Hz, 1H), 7.90 (brs, 1H), 7.87-8.01 (m, 1H), 7.62-7.74 (m, 1H), 7.23-7.33 (m, 2H), 7.17-7.21 (m, 1H), 6.38 (d, J=4.5 Hz, 1H), 3.34-3.40 (s, 6H), 2.91-3.06 (m, 4H), 2.12 (brt, J=7.0 Hz, 2H). MS ESI calculated for C.sub.22H.sub.22N.sub.6O.sub.3S.sub.3 [M+H].sup.+ 515, found 515.

Biological Test Data

Experimental Embodiment 1: IC.SUB.50 .Experiment for the Detection of NLRP3 Antagonist Using THP-1 Cells

[0173] The chemical names and structural formulas of the compounds of the present disclosure for experimental use are shown in the preparation embodiments for each compound.

[0174] 1. Experimental principle: In this experiment, the human monocytic cell line THP1 was used to study the inhibitory activity (IC.sub.50) of NLRP3 antagonists on the secretion of IL-1β. The monocytic cell line THP1 was differentiated into mature macrophages using PMA (phorbol 12-myristate 13-acetate), and then the cells were stimulated using LPS (lipopolysaccharide), an agonist of Toll-like receptor TLR4, to activate the transcriptional activity of inflammasome NLRP3 and the expression of IL-1β precursor pro-IL-1β. At this time, the NLRP3 antagonist was added, and then ATP was added to further mature and activate NLRP3, and activate downstream caspase-1. Pro-IL-1β could be enzyme-cleaved by the activated caspase-1 to mature IL-1β that could be secreted. The NLRP3 antagonist could effectively inhibit the maturation and activation of NLRP3 induced by ATP, as well as the activation of downstream caspase-1, thereby inhibiting the maturation and secretion of IL-1β.

[0175] 2. Experimental Materials:

[0176] 2.1 Reagents are Shown in Table 1:

TABLE-US-00001 TABLE 1 Item number or Storage Name Supplier serial number condition PMA Sigma 79346 −20° C. LPS InvivoGen tlrl-eblps −20° C. ATP — — −20° C. 1640 medium Gibco 22400-089  4° C. FBS HyClone SV30087.03 −80° C. Penicillin-streptomycin HyClone SV30010  4° C. β-Mercaptoethanol Sigma M3148 Room temperature NEAA non-essential Gibco 1140-050  4° C. amino acids Human soluble BD 558265 Room protein kit temperature Human IL-1β Flex Set BD 558279 Room temperature 96-well flat-bottom Corning 3599 Room plate temperature 96-well U-bottom Corning 3799 Room plate temperature

[0177] 2.2 Instruments are Shown in Table 2:

TABLE-US-00002 TABLE 2 Name Supplier Item number or serial number Flow Cytometer BD LSRFortessa

[0178] 2.3 Experimental Steps:

[0179] (1) The density of THP1 cells was adjusted to 5*10.sup.5 cells/mL, then PMA was added, and the final concentration was adjusted to 100 ng/mL, and the cells were inoculated into a 96-well flat-bottom plate with 200 μL/well, stimulated overnight at 37° C. and 5% CO.sub.2 (<16 hours if possible).

[0180] (2) The next day, the supernatant was discarded, and then carefully washed twice with Dulbecco's phosphate buffer (200 μL/time).

[0181] (3) The cells were stimulated with LPS, and the final concentration of LPS was 100 ng/mL, added to a 96-well plate with 200 μL/well, and cultured at 37° C. and 5% CO.sub.2 for 3 hours.

[0182] (4) The test compounds were added to the wells and the screening concentrations were respectively: 5 μM, 1 μM, 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM and 0.064 nM. The cells were incubated at 37° C. and 5% CO.sub.2 incubator for 1 hour.

[0183] (5) ATP was added to each well at a final concentration of 5 mM, and incubated overnight (>18 hours) at 37° C. and 5% CO.sub.2.

[0184] (6) On the third day, 5 μL of the supernatant was taken out, diluted 10 times, and the content of IL-1β in the supernatant was detected by CBA.

[0185] 3. Experimental Results:

[0186] The activity results of compounds are shown in Table 3.

TABLE-US-00003 TABLE 3 Results of NLRP3 antagonist inhibitory activity for compounds IL-1β inhibitory activity in THP-1 cell Compound IC.sub.50 (nM) 1 36.5 2 40.0 3 10.0 4a 33.9 5a 7.4 7a 9.8 8 27.8 — —

[0187] Experimental conclusion: The compounds of the present disclosure exhibits good NLRP3 inhibitory activity.

Experimental Embodiment 2: Pharmacokinetic Evaluation of Compounds

[0188] Experimental objective: To test the pharmacokinetics of compounds in mice

[0189] Experimental materials: C57BL/6J mice (male, 6-8 weeks old)

[0190] Experimental operation: The clear solution obtained after the test compound was dissolved was administered to female C57BL/6J mice (overnight fasting, 6-8 weeks old) via tail intravenous injection and intragastric administration (vehicle: 10% DMSO/10% solutol/80% water). After administration of test compound or control compound, blood was collected from the mandibular vein and centrifuged to obtain plasma at 0.0833, 0.25, 0.5, 1, 2, 4, 8 and 24 h for the intravenous injection group (IV) and at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h for the intragastric administration group (PO). The plasma concentration was determined by LC-MS/MS, and relevant pharmacokinetic parameters were calculated by the non-compartmental model linear logarithmic trapezoidal method using pharmacokinetic software WinNonlin™ Version 6.3. Meaning of each parameter: T.sub.1/2: half-life; C.sub.max: peak concentration; AUC.sub.0-inf: area under the plasma concentration-time curve from time 0 extrapolated to infinite time; F: bioavailability, Vd: apparent volume of distribution, Cl: clearance rate, T.sub.max: time to peak. The test results are shown in Table 4:

TABLE-US-00004 TABLE 4 Results of pharmacokinetic test of compound 7a Parameter Compound 7a IV T.sub.1/2 (h) 2.10 3 mg/kg Vd (L/kg) 0.25 Cl (mL/min/kg) 1.68 AUC.sub.0-inf (nM*h) 65725 PO: C.sub.max (nM) 101913 20 mg/kg T.sub.max (h) 0.5 AUC.sub.0-inf (nM*h) 296204 F (%) 67.6

[0191] Conclusion: The compounds of the present disclosure have good oral bioavailability, high exposure, and good pharmacodynamic properties in vivo.

Experimental Embodiment 3: Evaluation of the Therapeutic Effect of the Compound on the MSU-Induced Air Pouch Acute Gout Model in C57BL/6 Mice

[0192] Air Pouch of mouse is a cystic space similar to the human synovial membrane, and the injection of monosodium urate crystals (MSU) into the air pouch will cause an acute inflammatory response similar to human gout. The inflammatory cytokines IL-6 and IL-1β in the Air Pouch Lavage Fluid flushing fluid (APLV) were analyzed, and MCC950 was used as a control compound, and the efficacy of the compound of the present disclosure on the MSU-induced air pouch gout model in male C57BL/6 mice was tested.

[0193] Experimental objective: To evaluate the effect of the compound of the present disclosure on treating acute gout with Air Pouch gout model in mice.

[0194] Experimental animals: C57BL/6 mice, male, 7-8 weeks old, Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0195] Experimental Design:

[0196] As shown in FIG. 1, experimental healthy mice were numbered and grouped, and sterile air was injected into the back of the mice on the first day (Day 1) and on the fourth day (Day 4) to generate air pouch. On the seventh day, the drug was administered first, and the MSU crystal solution was injected into the air pouch after 1 hour, and the Air Pouch Lavage Fluid (APLV) was collected after 7 hours and analyzed. Grouping and administration scheme are shown in Table 5.

TABLE-US-00005 TABLE 5 Grouping and administration scheme Admini- Number stration of Im- dosage and Group animals munogen Test drug pathway Vehicle 1 5 None Navie — — 2 8 MSU Vehicle — — (3 mg) 3 8 MSU MCC950 50 mg/kg; 10% DMSO/10% (3 mg) po solutol/ 80% water 4 8 MSU Embodi- 50 mg/kg; 10% DMSO/10% (3 mg) ment po solutol/ 7a 80% water 5 8 MSU Embodi- 15 mg/kg; 10% DMSO/10% (3 mg) ment po, solutol/ 7a 80% water 6 8 MSU Embodi- 5 mg/kg; 10% DMSO/10% (3 mg) ment po solutol/ 7a 80% water 7 8 MSU Dex. 10 mg/kg; Physiological (3 mg) ip saline Note: Navie: healthy control group; Vehicle: vehicle control group; MCC950: reference compound; Dex.: dexamethasone; po: oral administration; ip: intraperitoneal injection.

[0197] Experimental Methods and Steps:

[0198] 1.1 Preparation of MSU

[0199] 1 g of uric acid was dissolved in 0.2 L of boiling water containing 6 mL of 1 N sodium hydroxide; after the pH was adjusted to 7.4, the solution was gradually cooled at room temperature and then left overnight at 4° C. MSU crystals were recovered by centrifugation and evaporated to dryness, dispensed into individual vials (3 mg), and sterilized by autoclaving.

[0200] 1.2 Grouping, Administration and Detection of IL-6 and IL-1β

[0201] Healthy C57BL/6 mice were numbered and grouped in the experiment, and 5 mL of sterile air was subcutaneously injected into the back of the mice on the same day of grouping (Day 1) and on the fourth day (Day 4) to generate an air pouch. On the seventh day (Day 7), each group of mice was given vehicle or test sample, and after 1 hour the suspension of MSU crystals (saline, 3 mg/mL) was injected into the air pouch. Air Pouch Lavage Fluid (APLV) would be collected after 6 hours, and ELISA kits would be used to test the levels of IL-6 and IL-1β in APLV. Results were expressed as average value±SEM. Statistical analysis was performed by a method of analysis of variance (ANOVA), followed by Dunnett test, and differences were considered significant when p<0.05.

[0202] Experimental Results

[0203] Compared with healthy control group, an acute inflammatory response in the air pouch of mice was induced by MSU injection, manifested by significantly increased concentrations of inflammatory cytokines IL-6 and IL-1β in APLV. The levels of IL-6 and IL-1β in APLV decreased rapidly after treatment with compound MCC950, compound 7a and dexamethasone. Among them, compound 7a was better than dexamethasone (10 mg/kg dose) in reducing IL-6 at high, medium and low doses, and better than MCC950 (50 mg/kg dose) in reducing IL-6 at the dose of 15 mg/kg and 50 mg/kg. Compound 7a had a significant effect on reducing IL-1β, and the effect on reducing IL-1β at the dose of 15 mg/kg and 50 mg/kg was significantly better than that of MCC950 (50 mg/kg dose), and with extremely low IL-1β levels, reaching the same effect as dexamethasone (10 mg/kg). Results of the inhibition experiment of the inflammatory cytokine IL-6 in APLV are shown in FIG. 1, and results of the inhibition experiment of the inflammatory cytokine IL-1β in APLV are shown in FIG. 2, and p represents significant difference, *: p<0.05; **: p<0.01; ***: p<0.001.

[0204] Conclusion: The compounds of the present disclosure have a good therapeutic effect on the MSU-induced Air Pouch gout model in C57BL/6 mice, and have the potential to treat gout and other diseases related to inflammatory cytokines.