SGLTS/DPP4 INHIBITOR AND APPLICATION THEREOF

Abstract

A class of compounds as a SGLT1/SGLT2/DPP4 triple inhibitor, and an application in preparation of a drug serving as the SGLT1/SGLT2/DPP4 triple inhibitor. Compounds represented by formula (I), and isomers and pharmaceutically-acceptable salts thereof are specifically involved.

##STR00001##

Claims

1. A compound of formula (I), an isomer or a pharmaceutically acceptable salt thereof, ##STR00063## wherein, R.sub.1 is selected from C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1, 2 or 3 R.sub.a; R.sub.2 is selected from Cl, Br, I, OH, NH.sub.2 and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1, 2 or 3 R.sub.b; R.sub.3, R.sub.4, R.sub.5, R.sub.6 and R.sub.7 are each independently selected from H, F, Cl, Br, I, OH, NH.sub.2 and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1, 2 or 3 R.sub.c; R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are each independently selected from H, F, Cl, Br, I, OH, NH.sub.2 and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1, 2 or 3 R.sub.d; E.sub.1 is —(CH.sub.2).sub.m—; E.sub.2 is —(CH.sub.2).sub.n—; m is 0, 1 or 2; n is 0, 1 or 2; R.sub.a, R.sub.b, R.sub.c and R.sub.d are each independently selected from F, Cl, Br, I, OH and NH.sub.2.

2. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.1 is selected from CH.sub.3 and Et, and the CH.sub.3 and Et are optionally substituted by 1, 2 or 3 R.sub.a.

3. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 2, wherein R.sub.1 is selected from CH.sub.3.

4. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.2 is selected from Cl, Br, I, OH, NH.sub.2, CH.sub.3, Et and ##STR00064## wherein the CH.sub.3, Et and ##STR00065## are optionally substituted by 1, 2 or 3 R.sub.b.

5. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 4, wherein R.sub.2 is selected from Cl, Br, I, OH, NH.sub.2, CH.sub.3, Et and ##STR00066##

6. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.3, R.sub.4, R.sub.5, R.sub.6 and R.sub.7 are each independently selected from H, F, Cl, Br, I, OH, NH.sub.2, CH.sub.3 and Et, wherein the CH.sub.3 and Et are optionally substituted by 1, 2 or 3 R.sub.c.

7. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 6, wherein R.sub.3, R.sub.4, R.sub.5, R.sub.6 and R.sub.7 are each independently selected from H, F, Cl, Br, I, OH and NH.sub.2.

8. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are each independently selected from H, F, Cl, Br, I, OH, NH.sub.2, CH.sub.3 and Et, wherein the CH.sub.3 and Et are optionally substituted by 1, 2 or 3 R.sub.d.

9. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 8, wherein R.sub.8, R.sub.9, R.sub.10 and R.sub.11 are each independently selected from H, F, Cl, Br, I, OH, NH.sub.2, CH.sub.3 and Et.

10. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein E.sub.1 is —CH.sub.2— or —CH.sub.2—CH.sub.2—.

11. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein E.sub.2 is a single bond or —CH.sub.2—.

12. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from ##STR00067## wherein, R.sub.1 is as defined in claim 1; R.sub.2 is as defined in claim 1; R.sub.3, R.sub.4, R.sub.5, R.sub.6 and R.sub.7 are as defined in claim 1; R.sub.8, R.sub.9, R.sub.10, and R.sub.11 are as defined in claim 1.

13. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 12, which is selected from ##STR00068## wherein, R.sub.1 is as defined in claim 12; R.sub.2 is as defined in claim 12; R.sub.3, R.sub.4, R.sub.5, R.sub.6 and R.sub.7 are as defined in claim 12; R.sub.8, R.sub.9, R.sub.10, and R.sub.11 are as defined in claim 12.

14. The following compound, an isomer or a pharmaceutically acceptable salt thereof, ##STR00069## ##STR00070##

15. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 14, which is selected from ##STR00071##

16. The compound, the isomer or the pharmaceutically acceptable salt thereof according to claim 15, which is selected from ##STR00072## ##STR00073##

17. A method for triple inhibition of SGLT1/SGLT2/DPP4, comprising administering a therapeutically effective amount of the compound or the pharmaceutically acceptable salt thereof according to claim 1 to a subject in need thereof.

Description

DETAILED DESCRIPTION

[0067] The present disclosure will be described in detail through the following examples, but it is not meant to impose any disadvantageous restriction on the present disclosure. The present disclosure has been described in detail herein, and its specific embodiments are also disclosed. For those skilled in the art, it is obvious to make various changes and improvements to the specific embodiment of the invention without departing from the spirit and scope of the invention.

REFERENCE EXAMPLE 1: FRAGMENTS A-1, A-2

[0068] ##STR00024##

Synthetic Route

[0069] ##STR00025##

Step 1: Synthesis of Compound A-1-2

[0070] Compound A-1-1 (25 g, 133.52 mmol, 1 eq) was dissolved in dichloromethane (110 mL), and added with triethylamine (27.02 g, 267.04 mmol, 37.17 mL, 2 eq). The reaction was cooled to 0° C., and methanesulfonyl chloride (15.30 g, 133.52 mmol, 10.33 mL, 1 eq) was added dropwise to the reaction at 0° C. After the addition was completed, the reaction temperature was raised to 15° C. and stirred at 15° C. for 3 hours. After the reaction was completed, the reaction was cooled to 0° C., and added slowly with water (100 mL) at 0° C. to quench the reaction. The mixture was extracted with dichloromethane (100 mL×2), the organic phase was combined and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain compound A-1-2.

Step 2: Synthesis of Compound A-1-4

[0071] A mixture of Compound A-1-2 (6.00 g, 22.61 mmol, 1.00 eq), compound A-1-3 (3.91 g, 22.61 mmol, 1.00 eq), and cesium carbonate (14.73 g, 45.22 mmol, 2.00 eq) in N,N-dimethylformamide (10.00 mL) was heated to 80° C., and stirred at 80° C. for 3 hours. After the reaction was completed, the reaction was added with water (50 mL). The mixture was extracted with ethyl acetate (30 mL×3), and the organic phases were combined and washed with water (30 mL×3) and saturated brine (30 mL) sequentially, dried with anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-1-4. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.41-7.34 (m, 2H), 6.78-6.74 (m, 2H), 4.15-4.12 (m, 1H), 3.64-3.52 (m, 4H), 2.18-2.07 (m, 2H), 1.157 (s, 9H).

Step 3: Synthesis of Compound A-1-5

[0072] Compound A-1-4 (3.00 g, 8.77 mmol, 1.00 eq) was dissolved in a solution of hydrogen chloride in ethyl acetate (10 mL, 4M). The reaction system was reacted at 20° C. for 1 hour. After the reaction was completed, the reaction was diluted with water (30 mL), and the mixture was washed with ethyl acetate (20 mL). The aqueous phase was adjusted to pH=7 with saturated aqueous sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (30 mL). The organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product of compound A-1-5. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.39-7.35 (m, 2H), 6.76-6.74 (m, 2H), 5.30 (m, 1H), 3.31-3.13 (m, 4H), 2.15-2.05 (m, 2H).

Step 4: Synthesis of Compounds A-1-7 and A-1-8

[0073] Compound A-1-5 (2 g, 7.18 mmol, 1 eq), compound A-1-6 (2.58 g, 7.90 mmol, 1.1 eq) were dissolved in dichloromethane (20 mL) and methanol (4 mL). The reaction was added with acetic acid (43.11 mg, 717.95 μmol, 41.06 μL, 0.1 eq) , followed by sodium triacetoxyborohydride (3.04 g, 14.36 mmol, 2 eq). The reaction was stirred at 15° C. for 2 hours. After the reaction was completed, the reaction was concentrated under reduced pressure, and the residue was diluted with water (50 mL) and extracted with dichloromethane (50 mL×2). The organic phases were combined and washed with saturated brine (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=3/1) to obtain compound A-1-7 (Rf=0.5) (.sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.38 (d, J=9.2 Hz, 2 H), 7.34-7.28 (m, 1 H), 7.03-6.87 (m, 2 H), 6.76 (d, J=8.8 Hz, 2 H), 4.93-4.77 (m, 1 H), 4.49-4.28 (m, 2 H), 4.15 (br d, J=12.4 Hz, 1 H), 4.10-3.97 (m, 1 H), 3.59 (d, J=12.0 Hz, 1 H), 3.50-3.35 (m, 1 H), 3.08-2.89 (m, 1 H), 2.76-2.56 (m, 2 H), 2.50 (br s, 1 H), 2.45-2.37 (m, 1 H), 2.36-2.26 (m, 1 H), 2.12-1.94 (m, 1 H), 1.28 (br s, 9 H)) and compound A-1-8 (Rf=0.4) respectively (.sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.37 (d, J=8.8 Hz, 2 H), 7.24-7.17 (m, 1 H), 7.02-6.89 (m, 2 H), 6.74 (d, J=8.8 Hz, 2 H), 4.85-4.74 (m, 1 H), 4.51-4.40 (m, 1 H), 4.31-4.25 (m, 1 H), 4.24-4.18 (m, 1 H), 3.82-3.65 (m, 1 H), 3.45-3.33 (m, 1 H), 3.04-2.83 (m, 3 H), 2.65-2.50 (m, 2 H), 2.50-2.41 (m, 1 H), 2.38-2.23 (m, 1 H), 2.06-1.93 (m, 1 H), 1.26 (br s, 9 H)).

Step 5: Synthesis of Compound A-1

[0074] Compound A-1-8 (15 g, 27.10 mmol, 1 eq), compound A-1-9 (13.77 g, 54.21 mmol, 2 eq), potassium acetate (7.98 g, 81.31 mmol, 3 eq), [1,1-bis(diphenylphosphorus)ferrocene]palladium dichloride (1.98 g, 2.71 mmol, 0.1 eq) were sequentially added to the anhydrous dioxane (20 mL), and reacted at 90° C. for 2 hours under protection of nitrogen. After the reaction was completed, the reaction was concentrated under reduced pressure, the residue was diluted with water (200 mL), and the mixture was extracted with dichloromethane (200 mL×2). The organic phases were combined and washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-1. .sup.1H NMR (400 MHz, CDCl.sub.3) δ7.74 (d, J=8.8 Hz, 2H), 7.24-7.15 (m, 1H), 7.01-6.90 (m, 2H), 6.85 (d, J=8.8 Hz, 2H), 4.93-4.83 (m, 1H), 4.46 (br d, J=9.6 Hz, 1H), 4.27 (br d, J=9.6 Hz, 1H), 4.24-4.18 (m, 1H), 3.81-3.68 (m, 1H), 3.38 (br t, J=10.8 Hz, 1H), 3.05-2.96 (m, 1H), 2.93 (br d, J=3.2 Hz, 2H), 2.63-2.49 (m, 2H), 2.46 (br d, J=11.6 Hz, 1H), 2.39-2.27 (m, 1H), 2.05-1.95 (m, 1H), 1.51 (q, J=12.0 Hz, 1H), 1.34 (s, 12H), 1.26 (s, 9H).

Step 6: Synthesis of Compound A-2

[0075] Compound A-1-7 (3.20 g, 5.78 mmol, 1 eq), compound A-1-9 (2.20 g, 8.67 mmol, 1.5 eq), potassium acetate (1.70 g, 17.35 mmol, 3 eq), [1,1-bis(diphenylphosphino)ferrocene]palladium(II)dichloride dichloromethane adduct (472.19 mg, 578.21 μmol, 0.1 eq) were added to anhydrous dioxane (30 mL) and the mixture was stirred at 90° C. for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was diluted with water (50 mL), and the mixture was extracted with dichloromethane (50 mL×2). The organic phases were combined and washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-2. .sup.1HNMR (400 MHz, CDCl.sub.3) δ 7.74 (d, J=8.4 Hz, 2H), 7.34-7.25 (m, 1H), 7.00-6.90 (m, 2H), 6.86 (d, J=8.4 Hz, 2H), 5.00-4.86 (m, 1H), 4.50-4.35 (m, 2H), 4.18-4.07 (m, 2H), 4.06-9.95 (m, 1H), 3.57 (br d, J=12.0 Hz, 1H), 3.54-3.44 (m, 1H), 3.08-2.93 (m, 1H), 2.72-2.63 (m, 1H), 2.62-2.54 (m, 1H), 2.43-2.36 (m, 1H), 2.35-2.26 (m, 1H), 2.04-1.96 (m, 1H), 1.33 (s, 12H), 1.23 (s, 9H).

REFERENCE EXAMPLE 2: Fragments A-3, A-4

[0076] ##STR00026##

Synthetic Route

[0077] ##STR00027##

Step 1: Synthesis of Compound A-3-2

[0078] Methanesulfonyl chloride (6.61 g, 57.73 mmol, 4.47 mL, 1.25 eq) was added to a mixture of compound A-3-1 (8 g, 46.19 mmol, 1 eq) and triethylamine (9.35 g, 92.37 mmol, 12.86 mL, 2 eq), in dichloromethane (120 mL), and stirred at 20° C. for 16 hours. After the reaction was completed, the mixture was slowly poured into 200 mL of ice water to quench the reaction. After reaction was quenched, the mixture was stirred for 10 minutes. The layers were separated by standing. The organic phase was separated and washed with water (50 mL), sodium hydroxide aqueous solution (1M, 50 mL), saturated brine (50 mL) sequentially, dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain the product compound A-3-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.18 (tt, J=6.7, 4.1 Hz, 1H), 4.30-4.22 (m, 2H), 4.09 (dd, J=4.1, 0.9 Hz, 1H), 4.07 (dd, J=4.1, 0.9 Hz, 1H), 3.05 (s, 3H), 1.38-1.47 (s, 9H).

Step 2: Synthesis of Compound A-3-3

[0079] A mixture of 4-bromophenol (5.78 g, 33.43 mmol, 1.2 eq), A-3-2 (7 g, 27.86 mmol, 1 eq) and cesium carbonate (18.15 g, 55.71 mmol, 2 eq) in N,N-dimethyl formaldehyde (20 mL) was stirred at 80° C. for 16 hours. After the reaction was completed, the reaction was quenched by water (200 mL), and the mixture was extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. Petroleum ether (60 mL) was added to the crude product, stirred at room temperature for 1 hour, and then filtered. The filtered cake was dissolved in dichloromethane and concentrated under reduced pressure to obtain compound A-3-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.39 (d, J=8.5 Hz, 2H), 6.63 (d, J=8.8 Hz, 2H), 4.89-4.79 (m, 1H), 4.29 (dd, J=9.8, 6.3 Hz, 2H), 4.00 (d, J=4.0 Hz, 1H), 3.98 (d, J=4.0 Hz, 1H), 1.40-1.52 (s, 9H).

Step 3: Synthesis of Compound A-3-4

[0080] Compound A-3-3 (11 g, 33.52 mmol, 1 eq) was added to a solution of hydrogen chloride in ethyl acetate (4M, 30 mL, 3.58 eq). The mixture was stirred at 20° C. for 16 hours. After the reaction was completed, the mixture was filtered. The filter cake was washed with ethyl acetate (30 mL) and then dissolved in dichloromethane (40 mL). The dichloromethane solution was concentrated under reduced pressure to obtain the hydrochloride salt of compound A-3-4.

Step 4: Synthesis of Compounds A-3-5 and A-3-6

[0081] A mixture of the hydrochloride salt of compound A-3-4 (4.2 g, 15.88 mmol, 1 eq), compound A-1-5 (7.79 g, 23.81 mmol, 1.5 eq), magnesium sulfate (19.11 g, 158.76 mmol, 10 eq) and triethyl amine (1.61 g, 15.88 mmol, 2.21 mL, 1 eq) in N,N-dimethylacetamide (90 mL) was stirred at 20° C. for 1 hour, sodium cyanoborohydride (3.99 g, 63.50 mmol, 4 eq) was added and the reaction was continuously stirred for 15 hours. After the reaction was completed, the reaction was diluted with water (100 mL), and filtrated. The filter cake was washed with water (20 mL), dissolved with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography to obtain compound A-3-5 (petroleum ether: ethyl acetate=3:1, Rf=0.31) (.sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.44-7.33 (m, 2H), 7.19 (m, 1H), 6.95 (m, 2H), 6.74-6.60 (m, 2H), 4.77 (br t, J=5.6 Hz, 1H), 4.45 (m, 1H), 4.25 (br d, J=10.0 Hz, 1H), 4.07-3.97 (m, 1H), 3.79 (m, 3H), 3.33-3.15 (m, 3H), 2.63 (m, J=10.8 Hz, 1H), 2.30 (br d, J=13.3 Hz, 1H), 1.34-1.21 (s, 9H)) and compound A-3-6 (petroleum ether: ethyl acetate=3:1, Rf=0.46) respectively (.sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.38 (br d, J=9.0 Hz, 3H), 6.94 (m, 2H), 6.78-6.61 (m, 2H), 4.78 (m, 1H), 4.37 (m, 2H), 4.07-3.88 (m, 4H), 3.55 (br d, J=12.8 Hz, 1H), 3.07 (br s, 2H), 2.53 (br s, 1H), 2.17 (br d, J=11.5 Hz, 1H), 1.28 (s, 9H)).

Step 5: Synthesis of Compound A-3

[0082] Compound A-3-5 (200 mg, 370.78 μmol, 1 eq), bis(pinacolato)diboron (235.39 mg, 926.94 μmol, 2.5 eq), [1,1-bis(diphenylphosphino)ferrocene]palladium(II)dichloride dichloromethane adduct (60.56 mg, 74.16 μmol, 0.2 eq) and potassium acetate (109.17 mg, 1.11 mmol, 3 eq) were added to anhydrous dioxane (5 mL) under nitrogen protection, and the mixture was reacted at 80° C. for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was quenched with water (10 mL), and extracted with dichloromethane (5 mL×2). The organic phases were combined and washed with water (5 mL) and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-3. MS: 587.2 [M+1].sup.+.

Step 6: Synthesis of Compound A-4

[0083] Compound A-3-6 (200 mg, 370.78 μmol, 1 eq), bis(pinacolato)diboron (112.99 mg, 444.93 μmol, 1.2 eq), [1,1-bis(diphenylphosphino)ferrocene]palladium(II)dichloride dichloromethane adduct (60.56 mg, 74.16 μmol, 0.2 eq), potassium acetate (72.78 mg, 741.55 μmol, 2 eq) were added to anhydrous dioxane (4 mL) under nitrogen protection, and the mixture was reacted at 80° C. for 16 hours under nitrogen protection. After the reaction was completed, the reaction was diluted with water (10 mL), and extracted with dichloromethane (10 mL×2). The organic phases were combined and washed with water (5 mL) and concentrated under reduced pressure to obtain a crude product. The crude product is purified by column chromatography to obtain compound A-4. MS: 587.1 [M+1].sup.+.

REFERENCE EXAMPLE 3: FRAGMENT A-5

[0084] ##STR00028##

Synthetic Route

[0085] ##STR00029##

Step 1: Synthesis of Compound A-5-2

[0086] A mixture of compound A-5-1 (4 g, 13.67 mmol, 1 eq), compound A-1-6 (6.71 g, 20.51 mmol, 1.5 eq), triethylamine (1.38 g, 13.67 mmol, 1.90 mL, 1 eq), and magnesium sulfate (16.46 g, 136.71 mmol, 10 eq) in N,N-dimethylacetamide (40 mL) was reacted at 20° C. for 1 hour, sodium cyanoborohydride (3.44 g, 54.68 mmol, 4 eq) was added to the reaction, and the reaction was continuously stirred for 15 hour. After the reaction was completed, the reaction was quenched with water (50 mL), and extracted with dichloromethane (20 mL×2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography to obtain the product A-5-2, .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.40-7.22 (m, 2H), 7.21 (m, 1H), 7.02-6.89 (m, 2H), 6.83-6.76 (m, 2H), 4.48 (br d, J=8.8 Hz, 1H), 4.27 (br dd, J=7.3, 3.0 Hz, 2H), 4.24-4.18 (m,1H), 3.70 (br d, J=8.0 Hz, 1H), 3.42 (t, J=10.9 Hz, 1H), 2.93-2.71 (m, 3H), 2.56-2.38 (m, 3H), 2.04-1.93 (m, 2H), 1.80 (m, 4.3 Hz, 2H), 1.70 (m, 1H), 1.50 (q, J=11.9 Hz, 1H), 1.32-1.18 (s, 9H).

Step 2: Synthesis of Compound A-5

[0087] Compound A-5-2 (7 g, 12.34 mmol, 1 eq), bis(pinacolato)diboron (7.83 g, 30.84 mmol, 2.5 eq), potassium acetate (2.42 g, 24.67 mmol, 2 eq), and [1,1-bis(diphenylphosphino)ferrocene]palladium(II)dichloride dichloromethane adduct (2.01 g, 2.47 mmol, 0.2 eq) were added to anhydrous dioxane (40 mL), and the mixture was reacted at 80° C. for 16 hours under the protection of nitrogen. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (20 mL), and washed with water (20 mL×3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain the product A-5 MS: 615.3[M+1].sup.+.

REFERENCE EXAMPLE 4: FRAGMENTS A-6, A-7

[0088] ##STR00030##

Synthetic Route

[0089] ##STR00031##

Step 1: Synthesis of Compound A-6-2

[0090] Compound A-6-1 (10 g, 53.41 mmol, 1 eq) was dissolved in dichloromethane (100 mL), and then triethylamine (10.81 g, 106.82 mmol, 14.87 mL, 2 eq) was added to the solution. Methanesulfonyl chloride (6.12 g, 53.41 mmol, 4.13 mL, 1 eq) was added dropwise to the reaction at 0° C., and the reaction was stirred at 15° C. for 3 hours. After the reaction was completed, the reaction was quenched with water (20 mL) at 0° C., and then extracted with dichloromethane (30 mL×2). The organic phases were combined and washed with saturated brine (20 mL), dried with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain A-6-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.22-5.17 (m, 1H), 3.66-3.39 (m, 4H), 2.99 (s, 3H), 2.27-2.04 (m, 2H), 1.40 (s, 9H).

Step 2: Synthesis of Compound A-6-3

[0091] Compound A-6-2 (6.5 g, 24.50 mmol, 1 eq), 4-bromophenol (5.09 g, 29.40 mmol, 1.2 eq), cesium carbonate (15.96 g, 49.00 mmol, 2 eq) were added to N,N-dimethyl formamide (70 mL). The mixture was stirred at 80° C. for 2 hours. After the reaction was completed, the mixture was added with water (500 mL),then extracted with ethyl acetate (500 mL×2). The organic phases were combined and washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, the filter was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=3:1) to obtain compound A-6-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.37 (br t, J=8.2 Hz, 1H), 7.31-7.26 (m, 2H), 4.82 (br s, 1H), 3.63-3.43 (m, 4H), 2.21-2.05 (m, 2H), 1.47 (br d, J=3.3 Hz, 9H).

Step 3: Synthesis of Compound A-6-4

[0092] A solution of hydrogen chloride in ethyl acetate (4M, 21.92 mL, 2 eq) was added to a solution of compound A-6-3 (15.00 g, 43.83 mmol, 1 eq) in ethyl acetate (100 mL), and the reaction was stirred at 15° C. for 2 hours. After the reaction was completed, the reaction was concentrated under reduced pressure. A mixed solution (petroleum ether: ethyl acetate=5:1, 100 mL) was added to the residue, and stirred for 2 hours. The mixture was filtered, and the filter cake was collected to obtain compound A-6-4. .sup.1H NMR (400 MHz, CD.sub.3OD) δ=7.48-7.44 (m, 2H), 6.98-6.93 (m, 2H), 5.21 (td, J=2.7, 5.8 Hz, 1H), 3.57-3.45 (m, 4H), 2.37 -2.29 (m, 2H).

Step 4: Synthesis of Compound A-6-5 and A-6-6

[0093] Acetic acid (86.23 mg, 1.44 mmol, 82.12 μL, 0.1 eq) was added to the solution of compound A-6-4 (4 g, 14.36 mmol, 1 eq), compound A-1-6 (5.17 g, 15.79 mmol, 1.1 eq) in dichloromethane (50 mL) and methanol (20 mL), followed by sodium triacetoxyborohydride (6.09 g, 28.72 mmol, 2 eq). The reaction was stirred at 15° C. for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The residue was added with water (50 mL), and the mixture was extracted with dichloromethane (50 mL×2). The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=3/1-2/1, containing one thousandth of triethylamine) to obtain compound A-6-5 (Rf=0.47) (.sup.1H NMR (400 MHz, DMSO) δ=7.44 (d, J=8.9 Hz, 2H), 7.21-7.07 (m, 3H), 6.91 (d, J=8.9 Hz, 2H), 6.83 (d, J=9.8 Hz, 1H), 4.94 (br s, 1H), 4.03 (q, J=7.1 Hz, 2H), 3.95-3.82 (m, 1H), 3.46 (br d, J=12.0 Hz, 1H), 3.00 (br dd, J=6.1, 10.0 Hz, 1H), 2.86-2.72 (m, 2H), 2.39-2.28 (m, 2H), 2.07 (br d, J=13.0 Hz, 1H), 1.91-1.82 (m, 1H), 1.77 (br t, J=11.7 Hz, 1H), 1.21-1.17 (m, 9H)) and compound A-6-6 (Rf=0.40) respectively (.sup.1H NMR (400 MHz, DMSO) δ=7.43 (d, J=8.9 Hz, 2H), 7.20-7.12 (m, 3H), 6.95-6.85 (m, 3H), 4.84 (br s, 1H), 4.13-4.04 (m, 1H), 3.67-3.56 (m, 1H), 3.33 (d, J=9.5 Hz, 1H), 3.17 (br t, J=10.6 Hz, 1H), 2.88-2.81 (m, 1H), 2.81-2.72 (m, 1H), 2.81-2.71 (m, 1H), 2.48-2.36 (m, 2H), 2.30-2.20 (m, 1H), 2.13 (br d, J=11.9 Hz, 1H), 1.78-1.69 (m, 1H), 1.53 (q, J=11.9 Hz, 1H), 1.30-1.11 (m, 9H)).

Step 5: Synthesis of Compound A-6

[0094] Compound A-6-5 (4.50 g, 8.13 mmol, 1 eq), bis(pinacolato)diboron (3.10 g, 12.20 mmol, 1.5 eq), potassium acetate (1.60 g, 16.26 mmol, 2 eq), and [1,1-bis(diphenylphosphorus)ferrocene] palladium dichloride (594.96 mg, 813.10 μmol, 0.1 eq) were added to anhydrous dioxane (50 mL) and the mixture was reacted at 90° C. for 2 hours under protection of nitrogen. After the reaction was completed, the reaction was concentrated under reduced pressure and the residue was diluted with water (50 mL), extracted with dichloromethane (50 mL×2). The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (dichloromethane/methanol=20:1) to obtain compound A-6. .sup.1H NMR (400 MHz, CDCl.sub.3) δ=7.71-7.54 (m, 2H), 7.24 (br s, 1H), 6.86 (br s,2H),6.79 (br d, J=8.4 Hz, 2H), 5.23 (s, 1H), 4.86 (br s, 1H), 4.34 (br s, 2H), 4.09 (br d, J=12.1 Hz, 1H), 3.50 (br d, J=12.3 Hz, 1H), 2.84 (br s, 1H), 2.75-2.59 (m, 2H), 2.41 (br s, 1H), 2.36-2.11 (m, 3H), 1.98 (br d, J=7.3 Hz, 1H), 1.26 (s, 9H), 1.23-1.17 (m, 12H).

Step 6: Synthesis of Compound A-7

[0095] Compound A-6-6 (3.40 g, 6.14 mmol, 1 eq), bis(pinacolato)diboron (2.34 g, 9.22 mmol, 1.5 eq), potassium acetate (1.21 g, 12.29 mmol, 2 eq) and [1,1-bis(diphenylphosphino)ferrocene]palladium(II)dichloride dichloromethane adduct (501.70 mg, 614.35 μmol, 0.1 eq) were added to anhydrous dioxane (50 mL), and the mixture was stirred at 90° C. for 2 hours under protection of nitrogen. After the reaction was completed, the reaction was concentrated under reduced pressure. The residue was diluted with water (50 mL), and extracted with dichloromethane (50 mL×2). The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (dichloromethane/methanol=20:1) to obtain compound A-7. .sup.1H NMR (400 MHz, CDCl.sub.3) δ=7.70-7.59 (m, 2H), 7.13 (br s, 1H), 6.87 (br s, 2H), 6.82-6.70 (m, 2H), 5.25-5.20 (m, 1H), 4.79 (br s, 1H), 4.39 (br d, J=8.9 Hz, 1H), 4.17 (br d, J=13.5 Hz, 1H), 3.66 (br d, J=8.3 Hz, 1H), 3.43-3.30 (m, 1H), 2.86 (br d, J=19.4 Hz, 3H), 2.51 (br s, 2H), 2.38 (br d, J=10.9 Hz, 1H), 2.24 (br d, J=6.6 Hz, 1H), 1.95 (br d, J=6.4 Hz, 1H), 1.26 (s, 9H), 1.18 (br s, 12H).

REFERENCE EXAMPLE 5: FRAGMENT A-8, A-9

[0096] ##STR00032##

Synthetic Route

[0097] ##STR00033##

Step 1: Synthesis of Compound A-8-1

[0098] Compound A-1-2 (11.5 g, 43.34 mmol, 1.00 eq), 2-methyl-4-bromophenol (8.11 g, 43.34 mmol, 1.00 eq) and potassium carbonate (11.98 g, 86.69 mmol, 2.00 eq) were added to N,N-dimethylformamide (100.00 mL). The reaction was stirred at 80° C. for 3 hours. After the reaction was completed, the mixture was added with water (500 mL), extracted with ethyl acetate (250 mL×2), and the organic phases were combine and washed with 1M aqueous sodium hydroxide solution (200 mL×2) and saturated brine (200 mL×2) sequentially, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-8-1. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.22-7.01 (m, 2H), 6.67-6.52 (m, 1H), 4.10-3.97 (m, 1H), 3.57-3.35 (m, 3H), 2.97 (s, 1H), 2.16-2.02 (m, 3H), 2.01-1.82 (m, 1H), 2.02-1.76 (m, 1H), 1.40 (s, 9H).

Step 2: Synthesis of Compound A-8-2

[0099] Compound A-8-1 (14.8 g, 33.57 mmol, 1.00 eq) was dissolved in a solution of hydrogen chloride in ethyl acetate (50 mL, 4M), and the reaction was carried out at 15° C. for 3 hours. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was added with 10 mL of petroleum ether and 1 mL of ethyl acetate. The mixture was stirred for 1 hour, and then filtered. The filter cake was collected to obtain compound A-8-2. .sup.1H NMR (400 MHz, DMSO-d6) δ 7.38-7.29 (m, 2H), 6.96 (d, J=8.8 Hz, 1H), 5.15 (s, 1H), 3.90 (br t, J=5.2 Hz, 1H), 3.44 (br s, 2H), 3.24 (br d, J=10.8 Hz, 1H), 2.24-2.17 (m, 1H), 2.15 (s, 3H), 2.14-2.08 (m, 1H).

Step 3: Synthesis of Compounds A-8-3 and A-8-4

[0100] Acetic acid (20.5 mg, 341.76 μmol, 20 μL, 0.1 eq) and sodium triacetoxyborohydride (1.45 g, 6.84 mmol, 2 eq) were added sequentially to a solution of compound A-8-2 (1 g, 3.42 mmol, 1 eq) and compound A-1-6 (1.23 g, 3.76 mmol, 1.1 eq) in dichloromethane (20 mL) and methanol (4 mL). The reaction was stirred at 15° C. for 16 hours. After the reaction was completed, the reaction was concentrated under reduced pressure. The residue was diluted with water (20 mL), and extracted with dichloromethane (15 mL×2). The organic phases were combined and washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (dichloromethane/ethyl acetate=10/1) to obtain compound A-8-3 (Rf=0.1) (.sup.1H NMR (400 MHz, CDCl.sub.3) δ=7.19-7.18 (m, 1H), 7.15 (br dd, J=2.4, 8.4 Hz, 2H), 6.87 (br s, 2H), 6.50 (d, J=8.4 Hz, 1H), 4.70 (br t, J=6.8 Hz, 1H), 4.43-4.33 (m, 1H), 4.22-4.12 (m, 2H), 4.08-4.02 (m, 1H), 3.74-3.65 (m, 1H), 3.06-2.99 (m, 1H), 2.84-2.71 (m, 2H), 2.62-2.50 (m, 2H), 2.38 (br d, J=12.8 Hz, 1H), 2.25-2.17 (m, 1H), 2.11 (s, 3H), 1.67-1.53 (m, 1H), 1.45-1.37 (m, 1H), 1.19 (s, 9H)) and compound A-8-4 (Rf=0.4) (.sup.1H NMR (400 MHz, CDCl.sub.3) δ=7.19 (br d, J=2.4 Hz, 1H), 7.17-7.13 (m, 1H), 7.19-7.12 (m, 1H), 6.89-6.85 (m,2H), 6.51 (d, J=8.8 Hz, 1H), 4.70 (br t, J=6.8 Hz, 1H), 4.44-4.33 (m,1H), 4.24-4.18 (m, 1H), 4.14 (td, J=2.4, 8.8 Hz, 1H), 3.70-3.63 (m, 1H), 3.30 (br t, J=10.8 Hz, 1H), 3.07-3.00 (m, 1H), 2.85-2.71 (m, 2H), 2.55 (br d, J=11.6 Hz, 2H), 2.39 (br d, J=13.8 Hz, 1H), 2.25-2.16 (m, 1H), 2.11 (s, 3H), 1.60 (br s, 1H), 1.43 (br d, J=11.2 Hz, 1H), 1.19 (s, 9H)) respectively.

Step 4: Synthesis of Compound A-8

[0101] Compound A-8-4 (600 mg, 1.06 mmol, 1 eq), bis(pinacolato)diboron (403 mg, 1.59 mmol, 1.5 eq), potassium acetate (156 mg, 1.59 mmol, 1.5 eq) and [1,1-bis(diphenylphosphorus)ferrocene] palladium dichloride (173 mg, 0.211 mmol, 0.2 eq) were added to anhydrous dioxane (10 mL), and the mixture was reacted at 80° C. for 18 hours under protection of nitrogen. After the reaction was completed, the mixture was filtered. The filtrate was diluted with water (15 mL) and then extracted with dichloromethane (20 mL×2). The organic phases were combined and washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-8. .sup.1H NMR (400 MHz, CDCl.sub.3) δ=7.56-7.52 (m, 2H), 7.13 (br s, 1H), 6.87 (br s, 2H), 6.63 (d, J=8.4 Hz, 1H), 4.79 (br t, J=6.8 Hz, 1H), 4.40 (br d, J=9.2 Hz, 1H), 4.25-4.09 (m, 1H), 4.05 (q, J=7.2 Hz, 2H), 4.08-4.02 (m, 1H), 3.29 (br t, J=10.8 Hz, 1H), 3.06-3.00 (m, 1H), 2.83-2.74 (m, 2H), 2.60-2.51 (m, 2H), 2.38 (br d, J=13.3 Hz, 1H), 2.42-2.34 (m, 1H), 2.27-2.20 (m, 1H), 2.13 (s, 3H), 1.26 (s, 12H), 1.19 (s, 9H).

Step 5: Synthesis of Compound A-9

[0102] Compound A-8-3 (300 mg, 528.67 μmol, 1 eq), bis(pinacolato)diboron (201.4 mg, 793.01 μmol, 1.5 eq) were dissolved in anhydrous DMF (5 mL), and potassium acetate (77.8 mg, 793.01 μmol, 1.5 eq), [1,1-bi s(diphenylphosphino)ferrocene]palladium(II)dichloride dichloromethane adduct (86.4 mg, 105.73 umol, 0.2 eq) were added sequentially under protection of nitrogen. The reaction was stirred at 90° C. for 16 hours under protection of nitrogen. After the reaction was completed, the reaction was diluted with water (15 mL), and the mixture was extracted with ethyl acetate (20 mL×2). The organic phases were combined and washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A-9. (MS:615.5[M+1].sup.+)

REFERENCE EXAMPLE 6: FRAGMENT B-1

[0103] ##STR00034##

Synthetic Route

[0104] ##STR00035##

Step 1: Synthesis of Compound B-1-2

[0105] Toluene (1200 mL), compound B-1-1 (200 g, 929.86 mmol, 1 eq), and tetrabutylammonium bromide (20.00 g, 62.04 mmol, 6.67 % eq) were added sequentially to a solution of sodium hydroxide (200.00 g, 5.00 mol, 5.38 eq) in water (400 mL) and the mixture was stirred for 0.5 hour. Allyl bromide (157.49 g, 1.30 mol, 112.49 mL, 1.4 eq) was slowly added to the mixture. The reaction temperature was raised to 45-50° C. and stirred for 15 hours. After the reaction was completed, the reaction was cooled and the mixture was left to stand and layered. The aqueous phase was separated and extracted with toluene (300 mL). The organic phases were combined, washed with saturated brine (300 mL×3), and concentrated under reduced pressure. The residue was concentrated under reduced pressure twice with toluene (200 mL) to obtain compound B-1-2. .sup.1H NMR (CDCl.sub.3) δ 7.52 (d, J=2.0 Hz, 1H), 7.37 (dd, J=2.4, 8.4 Hz, 1H), 7.08 (d, J=8.4 Hz, 1H), 6.05-5.87 (m, 1H), 5.42-5.17 (m, 2H), 4.57-4.43 (m, 2H), 4.06 (d, J=5.6 Hz, 2H), 2.72-2.57 (m, 2H), 1.21 (t, J=7.6 Hz, 3H).

Step 2: Synthesis of Compound B-1-4

[0106] A solution of n-butyllithium (2.5M, 4.31 mL, 1.1 eq) was added to a solution of compound B-1-2 (2.5 g, 9.80 mmol, 1 eq) in anhydrous tetrahydrofuran (40 mL) at −78° C. under protection of nitrogen, and the mixture was stirred for 0.5 hours to obtain solution A. A solution of Tert-butyl magnesium chloride (1M, 12.74)mL, 1.3 eq) was added to a solution of compound B-1-3 (3.21 g, 11.76 mmol, 1.2 eq) in anhydrous tetrahydrofuran (30 mL) at 0° C. under protection of nitrogen, and the mixture was stirred for 0.5 hours to obtain solution B. Solution B was added to solution A at −78° C., and the mixture was stirred and reacted at −78° C. for 0.5 hour. The reaction temperature was raised to 25° C. and stirred for 2 hours. After the reaction was completed, the reaction solution was slowly added to a saturated aqueous solution of ammonium chloride solution (5 mL) to quench the reaction, followed by addition of saturated brine (5 mL). The organic phase was separated after standing. The aqueous phase was extracted with ethyl acetate (10 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was added with n-heptane (10 mL) and stirred for 1 hour and filtered, and the filter cake was collected to obtain compound B-1-4. MS:363.2[M+1].sup.+

Step 3: Synthesis of Compound B-1-5

[0107] Cerium trichloride heptahydrate (3.39 g, 9.11 mmol, 865.45 μL, 1 eq) was added to a solution of compound B-1-4 (3.3 g, 9.11 mmol, 1 eq) in methanol (50 mL). Sodium borohydride (1.38 g, 36.42 mmol, 4 eq) was added to the reaction at 0° C. and stirred for 0.5 hour. The reaction temperature was raised to 25° C. and stirred for 1.5 hours. After the reaction was completed, saturated aqueous solution of ammonium chloride (10 mL) was added to the reaction at 0° C., and the mixture was stirred for 1 hour and concentrated under reduced pressure. The residue was added with ethyl acetate (30 mL), water (3 mL), and anhydrous magnesium sulfate (3 g), and the mixture was stirred for 0.5 hours and filtered. The filtrate was collected, and the filter cake was extracted with ethyl acetate (15 mL×3). The organic phases were combined, washed with water (10 mL×2) and saturated brine (10 mL×2) sequentially, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. 2.8 g of the crude product was dissolved in acetic acid (14 mL) and water (14 mL), and the reaction was heated to 95-100° C. and stirred for 16 hours. After the reaction was completed, the reaction was concentrated under reduced pressure. The residue was added with toluene (10 mL) and concentrated under reduced pressure. After repeating twice, a crude product was obtained. 2.5 g of the crude product was dissolved in pyridine (12 mL), and acetic anhydride (6.29 g, 61.66 mmol, 5.77 mL, 8 eq) was added at 0° C. The reaction was stirred at 10-20° C. for 16 hours. After the reaction was completed, water (5 mL) was added to the reaction to quench the reaction in ice bath. The mixture was concentrated under reduced pressure. The residue was added with 1M aqueous hydrochloric acid (5 mL) to neutralize the remaining pyridine, and extracted with ethyl acetate (30 mL×2). The organic phases were combined and washed with saturated brine (25 mL) and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product. The crude product was purified by column chromatography to obtain compound B-1-5. .sup.1H NMR (CDCl.sub.3) δ 7.33 (s, 1H), 7.30-7.20 (m, 1H), 7.20-7.15 (m, 1H), 6.05-5.90 (m, 1H), 5.40-5.05 (m, 5H), 4.65-4.55 (m, 1H), 4.52 (s, 2H), 4.13 (q, J=7.2 Hz, 1H), 4.05-4.00 (m, 2H), 2.66 (q, J=7.6 Hz, 2H), 2.15-1.95 (m, 9H), 1.83 (d, J=12.4 Hz, 3H), 1.19 (t, J=7.2 Hz, 3H).

Step 4: Synthesis of Compound B-1-6

[0108] Compound B-1-5 (1.4 g, 2.84 mmol, 1 eq), thiourea (757.32 mg, 9.95 mmol, 3.5 eq) were dissolved in dioxane (14 mL), and trimethylsilyl trifluoromethanesulfonate (2.53 g, 11.37 mmol, 2.05 mL, 4 eq) was added slowly to the reaction at 25° C. The reaction was heated to 60° C. and stirred for 1.5 hours. The reaction was cooled to 0° C. Diisopropylethylamine (2.94 g, 22.74 mmol, 3.96 mL, 8 eq) and methyl iodide (2.02 g, 14.21 mmol, 884.80 μL, 5 eq) were added to the reaction. The reaction was heated to 25° C. and stirred for 17 hours. After the reaction was completed, water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (15 mL×2). The organic phases were combined, dried with anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound B-1-6. MS:503.1[M+23].sup.+

Step 5: Synthesis of Compound B-1-7

[0109] A mixture of compound B-1-6 (9.48 g, 19.73 mmol, 1 eq), barbituric acid (5.05 g, 39.45 mmol, 2 eq), and tetrakistriphenylphosphine palladium (2.28 g, 1.97 mmol, 0.1 eq) in ethanol (95 mL) was heated to 65° C. and stirred for 16 hours under the protection of nitrogen. After the reaction was completed, the reaction was quenched by water (100 mL). The mixture was concentrated under reduced pressure. The residue was extracted with ethyl acetate (100 mL×3).

[0110] The organic phases were combined, dried with anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to obtain compound B-1-7. MS: 463.2 [M+23].sup.+

Step 6: Synthesis of Compound B-1

[0111] Phosphorus tribromide (4.40 g, 16.27 mmol, 1.54 mL, 0.5 eq) was added dropwise to a solution of compound B-1-7 (14.33 g, 32.53 mmol, 1 eq) in anhydrous tetrahydrofuran (145 mL) at 0° C. under the protection of nitrogen. The reaction was stirred at 20° C. for 16 hours. After the reaction was completed, 2M potassium carbonate aqueous solution (75 mL) was added dropwise to the mixture to quench the reaction at 0° C., and after stirring for 10 minutes, the mixture was left to stand and layered. The organic phase was separated and the aqueous phase was extracted with ethyl acetate (200 mL). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product. Methyl tert-butyl ether (40 mL) and n-hexane (60 mL) were added to the crude product, and the mixture was stirred at 25° C. for 16 hours and then filtered. The filter cake was washed with a mixed solvent (methyl tert-butyl ether: n-hexane=2:3, 50 mL) and dried to obtain compound B-1 (15 g, 27.10 mmol, 1 eq) .sup.1H NMR (CDCl.sub.3-d) δ 7.32-7.28 (m, 1H), 7.28-7.23 (m, 1H), 7.23-7.18 (m, 1H), 5.41-5.29 (m, 1H), 5.22 (t, J=10.0 Hz, 1H), 5.11 (t, J=10.0 Hz, 1H), 4.59-4.47 (m, 3H), 4.43 (br d, J=9.6 Hz, 1H), 2.75 (q, J=7.2 Hz, 2H), 2.20 (s, 3H), 2.10 (s, 3H), 2.02 (s, 3H), 1.84 (s, 3H), 1.27 (t, J=7.6 Hz, 3H).

[0112] Referring to the synthesis method of steps 1 to 6 in Reference Example 6, the fragments B-2, B-3, and B-4 in Table 1 were synthesized.

TABLE-US-00001 TABLE 1 Reference Examples 2-4 Reference Example Fragment Structure Confirmation 2 B-2 [00036] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.26 − 7.22 (m, 2 H), 7.19 − 7.15 (m, 1 H), 5.34 (q, J = 9.9 Hz, 1 H), 5.22 (t, J = 9.5 Hz, 1 H), 5.10 (t, J = 9.7 Hz, 1 H), 4.55 (d, J = 9.8 Hz, 1 H), 4.53 − 4.45 (m, 2 H), 4.42 (d, J = 9.8 Hz, 1 H), 2.39 (s, 3 H), 2.21 (s, 3 H), 2.10 (s, 3 H), 2.02 (s, 3 H), 1.84 (s, 3 H). 3 B-3 [00037] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.43 − 7.36 (m, 2 H), 7.29 (dd, J = 8.3, 2.0 Hz, 1 H) 5.44 − 5.33 (m, 1 H), 5.28 − 5.17 (m, 1 H), 5.07 (t, J = 9.7 Hz, 1 H), 4.66 − 4.49 (m, 3 H), 4.44 (d, J = 10.0 Hz, 1 H), 2.21 (s, 3 H), 2.11 (s, 3 H), 2.02 (s, 3 H), 1.87 (s, 3 H). 4 B-4 [00038] MS: 540.8 [M + 23].sup.+

EXAMPLE 1: WXD001

[0113] ##STR00039##

Synthetic Route

[0114] ##STR00040##

Step 1: Synthesis of Compound WXD001-1

[0115] Compound A-1 (3 g, 5.96 mmol, 1 eq), compound B-1 (4.29 g, 7.15 mmol, 1.2 eq), sodium carbonate (1.26 g, 11.92 mmol, 2 eq) and tetrakistriphenylphosphonium palladium (1.38 g, 1.19 mmol, 0.2 eq) were suspended in a mixed solvent of toluene (86 mL), ethanol (21.5 mL) and water (21.5 mL). The reaction was stirred at 50° C. for 16 hours under protection of nitrogen. After the reaction was completed, the reaction was concentrated under reduced pressure, and the residue was diluted with dichloromethane (200 mL) and washed with water (100 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. Ethanol (80 mL) was added to the crude product and stirred for 30 minutes and then filtered. The filter cake was washed with ethanol (10 mL×3) and dried to obtain compound WXD001-1.

Step 2: Synthesis of Compound WXD001-2

[0116] Compound WXD001-1 (3.3 g, 3.68 mmol, 1 eq) was dissolved in ethyl acetate (66 mL), and added with a solution of hydrogen chloride in ethyl acetate (4M, 66.00 mL, 71.76 eq). The reaction was stirred at 15° C. for 16 hours. After the reaction was completed, the reaction was concentrated under reduced pressure to obtain compound WXD001-2.

Step 3: Synthesis of Compound WXD001

[0117] Lithium hydroxide monohydrate (1.02 g, 24.39 mmol, 27 eq) and water (6 mL) were added to a mixture of compound WXD001-2 (760 mg, 0.91 mmol, 1 eq), methanol (6 mL) and tetrahydrofuran (3 mL). The mixture was stirred at 25° C. for 16 hours. After the reaction was completed, the reaction was diluted with water (10 mL) and extracted with ethyl acetate (10 mL×3). The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative high performance liquid chromatography to obtain the target compound WXD001. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.21 (s, 2H), 7.17-7.11 (m, 1H), 7.08-6.94 (m, 5H), 6.77 (d, J=8.8 Hz, 2H), 4.87-4.79 (m, 1H), 4.37 (d, J=9.6 Hz, 1H), 4.25-4.09 (m, 3H), 3.95 (s, 2H), 3.68-3.61 (m, 1H), 3.55-3.46 (m, 2H), 3.37 (br t, J=10.0 Hz, 1H), 3.10 (br dd, J=10.0 Hz, J=6.0 Hz, 1H), 2.88 (q, J=7.6 Hz, 1H), 2.75 (br d, J=7.6 Hz, 2H), 2.69-2.51 (m, 4H), 2.40 (br d, J=11.6 Hz, 1H), 2.27 (br dd, J=14.0 Hz, J=7.2 Hz, 1H), 2.18 (s, 3H), 2.07-1.95 (m, 1H), 1.42 (q, J=12.0 Hz, 1H), 1.15 (t, J=7.6 Hz, 3H).

[0118] Referring to the synthesis method of steps 1 to 3 in Example 1, each of Examples 2-12 in Table 2 below was synthesized. The structures in Table 2 also represent possible isomers.

TABLE-US-00002 TABLE 2 Each isomer of Examples 2-12 Ex- Frag- Com- amples mentA Fragment B pound Structure 2 A-2 [00041] WXD002 [00042] 3 A-1 [00043] WXD003 [00044] 4 A-1 [00045] WXD004 [00046] 5 A-3 [00047] WXD005 [00048] 6 A-4 [00049] WXD006 [00050] 7 A-5 [00051] WXD007 [00052] 8 A-6 [00053] WXD008 [00054] 9 A-7 [00055] WXD009 [00056] 10 A-8 [00057] WXD010 [00058] 11 A-9 [00059] WXD011 [00060] 12 A-1 [00061] WXD012 [00062]

[0119] The hydrogen spectrum and mass spectrum data of each example are shown in Table 3.

TABLE-US-00003 TABLE 3 Proton spectrum and mass spectrum data of each example Example Compound .sup.1H NMR MS m/z 1 WXD001 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 671.1 [M + 1].sup.+ 7.21 (s, 2H), 7.17-7.11 (m, 1H), 7.08-6.94 (m, 5H), 6.77 (d, J = 8.8 Hz, 2H), 4.87- 4.79 (m, 1H), 4.37 (d, J = 9.6 Hz, 1H), 4.25-4.09 (m, 3H), 3.95 (s, 2H), 3.68-3.61 (m, 1H), 3.55-3.46 (m, 2H), 3.37 (br t, J = 10.0 Hz, 1H) 3.10 (br dd, J = 10.0 Hz, J = 6.0 Hz, 1H), 2.88 (q, J = 7.6 Hz, 1H), 2.75 (br d, J = 7.6 Hz, 2H), 2.69- 2.51 (m, 4H), 2.40 (br d, J = 11.6 Hz, 1H), 2.27 (br dd, J = 14.0 Hz, J = 7.2 Hz, 1H), 2.18 (s, 3H), 2.07-1.95 (m, 1H), 1.42 (q, J = 12.0 Hz, 1H), 1.15 (t, J = 7.6 Hz, 3H). 2 WXD002 .sup.1H NMR (400 MHz,CD.sub.3OD) δ 671.3 [M + 1].sup.+ 1.09 (br s, 3H), 1.66 (br s, 1H), 2.04 (br s, 1H), 2.14 (br s, 3H), 2.36 (br d, J = 12.55 Hz, 2H), 2.45 (br s, 1H), 2.61 (br d, J = 7.03 Hz, 2H), 2.72 (br s, 1H), 2.86 (br d, J = 10.29 Hz, 1H), 2.93 (br s, 1H), 3.16 (br s, 2H), 3.37-3.50 (m, 3H), 3.62 (br d, J = 11.54 Hz, 1H), 3.96 (br s, 2H), 4.06-4.25 (m, 2H), 4.37 (br t, J = 11.17 Hz, 2H), 6.79 (br d, J = 7.78 Hz, 2H), 7.03-7.23 (m, 7H), 7.50 (br s, 1H). 3 WXD003 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 657.1 [M + 1].sup.+ 7.17 (s, 2H), 7.15-7.11 (m, 1H), 7.08-6.94 (m, 5H), 6.78 (br d, J = 8.0 Hz, 2H), 4.84 (br s, 1H), 4.38 (br d, J = 9.6 Hz, 1H), 4.25-4.12 (m, 3H), 3.92 (s, 2H), 3.70-3.66 (t, J = 8.8 Hz, 1H), 3.51 (dd, J = 15.6 Hz, J = 8.8 Hz, 2H), 3.38 (t, J = 10.8 Hz, 1H), 3.12 (dd, J = 10.0 Hz, J = 6.0 Hz, 1H), 2.93-2.85 (m, 1H), 2.81- 2.73 (m, 2H), 2.72-2.64 (m, 1H), 2.63-2.53 (br s, 1H), 2.40 (br d, J = 11.6 Hz, 1H), 2.33- 2.28 (m, 1H), 2.27 (s, 3H), 2.19 (s, 3H), 2.03-2.00 (m, 1H), 1.44 (q, J = 12.0 Hz, 1H), 4 WXD004 .sup.1H NMR (400 MHz, DMSO-d.sub.6) 677.2 [M + 1].sup.+ δ 8.20 (s, 2H), 7.38 (d, J = 8.4 Hz, 1H), 7.28 (s, 1H), 7.27-7.18 (m, 4H), 7.10 (br d, J = 8.8 Hz, 2H), 6.80 (br d, J = 8.5 Hz, 2H), 4.587-4.76 (m, 1H), 4.34 (br d, J = 9.6 Hz, 1H), 4.19 (br d, J = 9.6 Hz, 1H), 4.09 (br d, J = 9.6 Hz, 1H), 4.06-4.03 (m, 1H), 3.98 (br s, 2H), 3.27 (br d, J = 8.4 Hz, 1H), 3.18 (dt, J = 18.0, 9.2 Hz, 4H), 3.00- 2.88 (m, 3H), 2.82-2.75 (m, 1H), 2.68 (br d, J = 8.0 Hz, 1H), 2.35-2.17 (m, 2H), 2.03 (s, 3H), 1.78-1.69 (m, 1H), 1.39 (q, J = 11.6 Hz, 1H). 5 WXD005 .sup.1H NMR (400 MHz, CD.sub.3OD) δ 663.1 [M + 1].sup.+ 7.36 (d, J = 8.0 Hz, 1H), 7.31- 7.23 (m, 3H), 7.22-7.16 (m, 2H), 7.13 (br d, J = 8.3 Hz, 2H), 6.74 (br d, J = 8.3 Hz, 2H), 4.87-4.80 (m, 1H), 4.51 (br d, J = 8.5 Hz, 1H), 4.39 (d, J = 9.5 Hz, 1H), 4.14 (d, J = 9.5 Hz, 1H), 4.01- 4.11 (m, 3H), 3.87 (m, 2H), 3.42-3.48 (m, 1H), 3.40-3.32 (m, 4H), 3.24 (br t, J = 10.8 Hz, 1H), 2.77 (m, 1H), 2.37 (m, 1H), 2.13 (s, 3H), 1.44 (m, 1H). 6 WXD006 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 663.1 [M + 1].sup.+ 7.64-7.51 (m, 1H), 7.40 (br d, J = 7.5 Hz, 1H), 7.34-7.26(m, 2H), 7.25-7.12 (m, 4H), 6.81 (br d, J = 8.3 Hz, 2H), 4.64 (br d, J = 9.8 Hz, 1H), 4.43 (br d, J = 9.5 Hz, 1H), 4.18 (br d, J = 9.8 Hz, 1H), 4.06 (m, 5H), 3.71 (br d, J = 12.3 Hz, 1H), 3.57 (m, 1H), 3.53-3.46 (m, 1H), 3.44-3.37 (m, 3H), 2.81 (br s, 1H), 2.85-2.76 (m, 1H), 2.32 (br d, J = 11.8 Hz, 1H), 2.17 (s, 3H), 1.99-1.85 (m, 1H). 7 WXD007 .sup.1H NMR (400 MHz, CD.sub.3OD) δ 685.3 [M + 1].sup.+ 7.32-7.25 (m, 1H), 7.24-7.13 (m, 5H), 7.05 (d, J = 8.5 Hz, 2H), 6.83 (d, J = 8.5 Hz, 2H), 4.48 (d, J = 9.8 Hz, 1H), 4.39 (d, J = 9.5 Hz, 2H), 4.26 (br d, J = 8.3 Hz, 1H), 4.13 (d, J = 9.0 Hz, 1H), 3.97 (s, 2H), 3.55 (t, J = 10.9 Hz, 1H), 3.50-3.34 (m, 4H), 2.96 (m, 3H), 2.73-2.56 (m, 4H), 2.49 (br d, J = 12.8 Hz, 1H), 2.14 (s, 3H), 2.02 (m, 2H), 1.82 (m, 2H), 1.72 (q, J = 12.3 Hz, 1H), 1.09 (t, J = 7.5 Hz, 3H). 8 WXD008 .sup.1H NMR (400 MHz, CD.sub.3OD) δ 671.3 [M + 1].sup.+ 7.46 (m, 1H), 7.27-7.13 (m, 5H), 7.07 (d, J = 8.5 Hz, 2H), 6.82 (d, J = 8.5 Hz, 2H), 4.96 (m, 1H), 4.66 (br d, J = 8.8 Hz, 1H), 4.38 (d, J = 9.5 Hz, 1H), 4.25 (br d = J = 13.1 Hz, 1H), 4.12 (d, J = 9.0 Hz, 1H), 3.97 (s, 2H), 3.72 (m, 2H), 3.49-3.32 (m, 3H), 3.18 (m, 1H), 3.05 (br s, 2H), 2.85 (m, 1H), 2.69 (m, 1H), 2.61 (q, J = 7.5 Hz, 2H), 2.51 (br d, J = 10.3 Hz, 1H), 2.40 (dq, J = 13.8, 7.1 Hz, 1H), 2.14 (s, 3H), 2.10 (m, 1H), 1.96 (m, 1H), 1.09 (t, J = 7.5 Hz, 3H). 9 WXD009 .sup.1H NMR CD.sub.3OD) δ 7.34-7.11 671.1 [M + 1].sup.+ (m, 8H), 6.90 (d, J = 8.4 Hz, 2H), 5.21 (br s, 1H), 4.71 (d, J = 10.0 Hz, 1H), 4.50 (br d, J = 6.0 Hz, 1H), 4.41 (d, J = 9.6 Hz, 1H), 4.15 (d, J = 9.2 Hz, 1H), 4.05- 3.95 (m, 2H), 3.85-3.58 (m, 7H), 3.51-3.36 (m, 3H), 2.82 (br d, J = 9.6 Hz, 1H), 2.62 (q, J = 7.6 Hz, 2H), 2.49 (br s, 1H), 2.55- 2.43 (m, 1H), 2.40-2.29 (m, 1H), 2.16 (s, 3H), 1.11 (t, J = 7.6 Hz, 3H). 10 WXD010 .sup.1H NMR (400 MHz, DMSO) δ = 685.4 [M + 1].sup.+ 7.28-7.04 (m, 6), 6.95-6.82 (m, 2H), 6.95-6.81 (m, 1H), 5.30- 5.05 (m, 2H), 4.92-4.73 (m, 2H), 4.32 (br d, J = 9.3 Hz, 1H), 4.12- 3.99 (m, 3H), 3.86 (br s, 2H), 3.27-3.14 (m, 2H), 3.02-2.95 (m, 1H), 2.75 (br d, J = 8.0 Hz, 2), 2.66-2.51 (m, 6H), 2.42 (br s, 2H), 2.26-2.14 (m, 2H), 2.11- 1.98 (m, 6H), 1.76 (br s, 1H), 1.41-1.21 (m, 2H), 1.06 (br t, J = 7.3 Hz, 4H). 11 WXD011 .sup.1H NMR (400 MHz, CD.sub.3OD) 685.5 [M + 1].sup.+ δ = 7.52-7.34 (m, 2H), 7.25- 7.12 (m, 1H), 7.22-7.09 (m, 3H), 6.97-6.85 (m, 2H), 6.76 (d, J = 8.4 Hz, 1H), 4.94 (br d, J = 6.8 Hz, 2H), 4.49 (d, J = 9.6 Hz, 1H), 4.40 (d, J = 9.6 Hz, 1H), 4.23 (br d, J = 12.8 Hz, 1H), 4.16-4.11 (m, 1H), 3.94 (s, 2H), 3.68 (br d, J = 11.6 Hz, 1H), 3.54-3.36 (m, 5H), 3.24 (br dd, J = 6.0, 10.8 Hz, 1H), 3.00 (q, J = 7.2 Hz, 1H), 2.90 (br d, J = 10.4 Hz, 1H), 2.84-2.76 (m, 1H), 2.63 (q, J = 7.6 Hz, 2H), 2.55 (br s, 1H), 2.50-2.32 (m, 2H), 2.16 (s, 6H), 2.14-2.14 (m, 1H), 1.84-1.74 (m, 1H), 1.48 (br d, J = 6.4 Hz, 1H), 1.36-1.19 (m, 1H), 1.12 (t, J = 7.6 Hz, 3H). 12 WXD012 .sup.1H NMR (400 MHz, CD.sub.3OD) δ 685.3 [M + 1].sup.+ 7.29-7.13 (m, 6H) 7.04 (d, J = 8.5 Hz, 2H) 6.78 (d, J = 8.5 Hz, 2H) 4.62 (br s, 1H) 4.46 (d, J = 9.8 Hz, 1H) 4.40 (d, J = 9.5 Hz, 1H) 4.24 (br dd, J = 11.04, 2.8 Hz, 1H) 4.13 (d, J = 9.0 Hz, 1H) 3.99 (s, 2H) 3.52-3.34 (m, 4H) 3.28-3.10 (m, 2H) 3.07-2.91 (m, 3H) 2.75-2.58 (m, 2H) 2.49 (br d, J = 10.0 Hz, 1H) 2.37- 2.26 (m, 1H) 2.15 (s, 3H) 2.06- 1.87 (m, 1H) 1.64 (q, J = 11.8 Hz, 1H) 1.09 (dd, J = 6.78, 3.0 Hz, 6H).

EXAMPLE 1. CELL ACTIVITY TEST IN VITRO

Experimental Steps and Methods

Biological Activity Experiment 1: SGLT1 Glucose Transport Test

1. Experimental Purpose

[0120] The effect of the compound on the glucose transport activity of the SGLT1 transporter was detected through measuring the amount of [.sup.14C]-labeled glucose entering the cells with high expression of Human-SGLT1.

2. Experimental Method

2.1. Cell Preparation

[0121] The cells stably expressing Human-SGLT1 used in the experiment were constructed by Shanghai WuXi AppTec. The SGLT1 cells were plated on a Cytostar-T (PerkinElmer) 96-well cell culture plate, and cultured overnight at 5% CO.sub.2 at 37° C.

2.2. SGLT1 Glucose Transport Test

[0122] 1) Experimental buffer: 10 mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES), 1.2 mM magnesium chloride (MgCl.sub.2), 4.7 mM potassium chloride (KCl), 2.2 mM calcium chloride (CaCl.sub.2) and 120 mM sodium chloride (NaCl).

[0123] 2) The compound was diluted with 100% dimethyl sulfoxide (DMSO) with 1 mM as the starting concentration and 8 points of 5-fold serial dilutions.

[0124] 3) 3 μM [.sup.14C]-labeled methyl α-D-glucopyranoside was prepared with experimental buffer.

[0125] 4) The cells were treated with 49 μL of experimental buffer, 1 μL of the compound which was gradient diluted, and 50 μL of 3 M [.sup.14C] isotope-labeled sugar solution at 37° C. for 2 hours.

[0126] 5) The isotope detector (Micro beta Reader) was used to read.

[0127] 6) The data was calculated by GraphPad Prism 5.0 software: log(inhibitor) vs. response—Variable slope to obtain the IC.sub.50 value of the test compound. The experimental results were shown in Table 3.

Biological Activity Experiment 2: SGLT2 Glucose Transport Test

1. Experimental Purpose

[0128] The effect of the compound on the glucose transport activity of the SGLT2 transporter was detected through measuring the amount of [.sup.14C]-labeled glucose entering the cells with high expression of Human-SGLT2.

2. Experimental Method

2.1. Cell Preparation

[0129] The cells stably expressing Human-SGLT2 used in the experiment were constructed by Shanghai WuXi AppTec. The SGLT2 cells were plated on a 96-well cell culture plate (Greiner) and cultured overnight at 5% CO.sub.2 at 37° C.

2.2. SGLT2 Glucose Transport Test

[0130] 1) Experimental buffer: 10 mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES), 1.2 mM magnesium chloride (MgCl.sub.2), 4.7 mM potassium chloride (KCl), 2.2 mM calcium chloride (CaCl.sub.2) and 120 mM sodium chloride (NaCl).

[0131] 2) Termination buffer: 10 mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES), 1.2 mM magnesium chloride (MgCl.sub.2), 4.7 mM potassium chloride (KCl), 2.2 mM calcium chloride (CaCl.sub.2), 120 mM sodium chloride (NaCl) and 1 μM LX4211.

[0132] 3) The compound was diluted with 100% dimethyl sulfoxide (DMSO) with a starting concentration of 10 μM and 8 points of 5-fold serial dilution.

[0133] 4) 6 μM [.sup.14C]-labeled methyl α-D-glucopyranoside was prepared with experimental buffer.

[0134] 5) The cells were treated with 49 μL of experimental buffer, 1 μL of gradient diluted compound, and 50 μL of 6 μM [.sup.14C] isotope-labeled sugar solution at 37° C. for 2 hours.

[0135] 6) The liquid in the well was aspirated and the cells were rinsed 3 times with termination buffer.

[0136] 7) The cells were lysed with 50 μL of 10% sodium hydroxide solution, the cell lysate was aspirated into the scintillation tube, and 2 mL of scintillation fluid was added.

[0137] 8) An isotope detector (Tricarb) was used to read.

[0138] 9) The data was calculated by GraphPad Prism 5.0 software: log(inhibitor) vs. response—Variable slope to obtain the IC.sub.50 value of the test compound. The experimental results were shown in Table 3.

Biological Activity Experiment 3: rhDPP4 Inhibitor Screening Experiment

1. Experimental Purpose

[0139] The inhibitory activity of the compounds on recombinant human dipeptidyl peptidase 4 (rhDPP4) was evaluated by measuring the median inhibitory concentration (IC.sub.50) value of the compounds. In this experiment, rhDPP4 was used to catalyze a substrate to produce luciferin, and reacted with luciferase to produce light signals, wherein the substrate is the luminescent precursor glycine-proline-aminofluorescein (Gly-Pro-aminoluciferin). The intensity of the light signal is directly proportional to the enzyme activity.

2. Experimental Method

[0140] 1) 250 nL of gradient diluted compound (4 times dilution, 10 detection concentrations) was transfered to a 384-well plate (PerkingElmer-6007299) using a non-contact nano-upgraded acoustic pipetting system (ECHO). The concentration of dimethylsulfoxide (DMSO) concentration was 0.5% in the final reaction system. Blank control wells (containing DMSO, substrate and 10 mM of tris(hydroxymethyl)aminomethane hydrochlorid (Tris-HCl)) and positive control wells (containing DMSO, substrate and rhDPP4) were set.

[0141] 2) The pre-packed frozen buffer containing luciferase was taken out and restored to room temperature, and then added with substrate to prepare a working solution with a substrate concentration of 20 μM. The rhDPP4 was prepared as a 0.2 ng/mL working solution with 10 mM Tris-HCl (pH8.0) aqueous solution.

[0142] 3) 25 μL of working solution containing 20 μM substrate and 25 μL of working solution containing 0.2 ng/mL rhDPP4 were added to the 384-well plate with the compound added already. The plate was centrifuged at 1000 rpm for 30 s. The plate was sealed with aluminum foil sealing film and incubated at room temperature for 1 hour.

[0143] 4) The light signal intensity was detected with a multi-function enzyme label detector EnVision. The raw data was used to calculate the inhibition activity of the compounds on rhDPP4.

[0144] Inhibitory activity %=100-(compound well signal value-blank control well signal value)/(positive control well signal value-blank control well signal value)*100; the inhibition percentage was imported into GraphPad Prism software for data processing to obtain the corresponding dose-effect curve and the IC.sub.50 value of the test compound was obtained. The experimental results were shown in Table 4:

TABLE-US-00004 TABLE 4 In vitro cell viability test results Human-SGLT1 Human-SGLT2 rhDPP4 Compound IC.sub.50 (nM) IC.sub.50 (nM) IC.sub.50 (nM) WXD001 3.46 2.0 34.88 WXD002 1.22 0.76 4217 WXD003 44.05 2.9 24.52 WXD004 137.5 3.9 44.8 WXD005 70.73 0.66 111.7 WXD006 13.63 1.41 NA WXD007 3.37 1.44 510.5 WXD008 8.13 1.21 NA WXD009 25.94 1.90 295.7 WXD010 144.4 21.17 164.5 WXD011 44.93 10.29 NA WXD012 11.16 6.14 317.1 Note: NA means no relevant data.

[0145] Conclusion: The compound of the present disclosure exhibits excellent inhibitory activity on Human-SGLT1, Human-SGLT2 and rhDPP4 in vitro.

EXAMPLE 2: DMPK Study In Vivo

[0146] Experimental purpose: Male C57 mice were used as test animals to determine the blood concentration of the compound and evaluate the pharmacokinetic behavior after a single administration.

[0147] Experimental operation: 6 healthy adult male C57 mice were selected, 3 for the intravenous injection group and 3 for the oral group. The test compound was mixed with an appropriate amount of solvent.sup.# of the intravenous injection group, and the mixture was vortexed and sonicated to prepare 1 mg/mL of a clear solution, which was filtered with a microporous membrane for later use. For oral group, the test compound was mixed with the solvent, and the mixture was vortexed and sonicated to prepare 1 mg/mL of a clear solution. After intravenous administration of 1 mg/kg or oral administration of 10 mg/kg to mice, whole blood was collected for a certain period of time, and plasma was prepared. The drug concentration was analyzed by LC-MS/MS method, and the pharmacokeneties parameters were calculated by Phoenix WinNonlin software (Pharsight, USA).

[0148] The experimental results are shown in Table 5:

TABLE-US-00005 TABLE 5 PK test results of the compounds Oral DNAUC Vd.sub.ss Cl Compound C.sub.max (nM) F % (nM.h/mpk) (L/kg) (mL/min/kg) T.sub.1/2 (h) WXD001 225 18.2 109 17.8 42.0 8.99 WXD004 171 10.2 113 13.2 20.7 13.4 Remarks: Cmax is the maximum concentration; F % is the oral bioavailability; Oral DNAUC = AUC.sub.PO/Dose, AUC.sub.PO is the oral exposure, Dose is the drug dose; Vd.sub.ss is the volume of distribution; Cl is the clearance rate; T.sub.1/2 is the half-life. ″#″: The solvent used in WXD001 was 10% N-methylpyrrolidone/10% polyethylene glycol-15 hydroxystearate/80% H2O;

[0149] The solvent used in WXD004 was 20% polyethylene glycol 400/10% polyethylene glycol-15 hydroxystearate/70% H.sub.2O.

[0150] Conclusion: The compound of the present disclosure showed certain oral exposure and bioavailability in mice.

EXAMPLE 3. DRUG EFFICACY STUDY OF RAT ORAL GLUCOSE TOLERANCE TEST (OGTT) IN VIVO

Experiment Summary

1. Animal

[0151]

TABLE-US-00006 Category SD rat Male Weeks/ ~8 weeks/ Gender Shanghai Animal weight 250 g Supplier Slack animal feed: Common rat and mice feed

2. Experiment Grouping

[0152]

TABLE-US-00007 No. of Compound Frequency of Mode of amimal Group grouping Dosage administration administration per group 1 Solvent 0 Single Gavage 5 control administration group 2 Positive 10 Single Gavage 5 compound mg/kg administration (Soagliflozin) 3 Test 10 Single Gavage 5 compound mg/kg administration

Experiment Process

1. Animal Adaptation and Preparation

[0153] The experimental animals needed to be acclimatized in the animal room for 1 week after arriving at the facility.

2. Fasting and Administration

[0154] The animals were fasted for 18 h in the metabolic cage, and drugs or solvents (2 mL/kg) were given according to the above table, and then 50% glucose solution (2 g/kg, 4 mL/kg) was administrated immediately.

3. Urine and Blood Sugar Test

[0155] Two hours after the animals were given sugar, they resumed eating. The time points of 0 min, 15 min, 30 min, 45 min, 60 min and 120 min were collected for blood glucose determination; urine during 0-24 h was used for urine sugar (mg/200 g) and urine volume test respectively.

4. Data Analysis

[0156] All values will be expressed as average values. Statistical analysis was evaluated using Graphpad Prism 6 one-way analysis of variance Tukey's multiple comparison test. P value of less than 0.05 was considered of statistically significant.

[0157] The experimental results are shown in Table 6:

TABLE-US-00008 TABLE 6 Experimental results of glucose tolerance in rats Solvent Positive control compound Compound group (Soagliflozin) WXD001 OGTT glucose level 1079  917****   670**** AUC.sub.0-2 hr (mol/L × min) Urine sugar level 0.4 2913**** 16.2 (mg/200 g BW) Urine volume 16.8  32.2**** 11.2 (mL/200 g BW) *means p < 0.5, **means p < 0.01, ***means p < 0.001, ****means p < 0.0001 vs. solvent control group. Remarks: 200 g BW is 200 g average weight. Conclusion: Compared with the solvent control group, the compound of the present disclosure can significantly reduce the blood glucose AUC level of the animal within 2 hours; the 24-hour urine glucose excretion level of the animals is lower than that of the positive compound.