Optically active 2-hydroxy tetrahydrothienopyridine derivatives, preparation method and use in manufacture of medicament thereof

Abstract

Optically active 2-hydroxytetrahydrothienopyridine derivatives represented by Formula I and pharmaceutically acceptable salts, preparation method and use in the manufacture of a medicament thereof are disclosed. The pharmacodynamic experiment results show that the present compounds of Formula I are useful for inhibiting platelet aggregation. The pharmacokinetic experiment results show that the present compound of Formula I can be converted in vivo into pharmacologically active metabolites and are therefore useful for inhibiting platelet aggregation. Therefore, the present compounds are useful for the manufacture of a medicament for preventing or treating thrombosis and embolism related diseases.

Claims

1. A method for preparing an optically active 2-hydroxytetrahydro thienopyridine derivative, wherein said derivative is (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate of formula (I) or a pharmaceutically acceptable salt thereof, comprising the following steps: (1) reacting an optically active compound of Formula II ##STR00036## with a compound of Formula III ##STR00037## or a salt thereof in the presence of a base, to obtain an optically active compound of Formula IV ##STR00038## or a salt thereof, wherein R.sup.7 is a C.sub.1-6 alkyl, trifluoromethyl, pentafluoroethyl, heptafluoropropyl, phenyl, or a Z-substituted phenyl, in which Z is a C.sub.1-3 alkyl, halo, cyano, nitro, or trifluoromethyl, and is at position 2, 3 or 4 of the phenyl ring; and (2) reacting the compound of Formula IV or the salt thereof with a compound of Formula V ##STR00039## or a compound of Formula VI ##STR00040## in the presence of a base, to obtain the compound of Formula I, ##STR00041##

2. A method for preparing an optically active 2-hydroxytetrahydro thienopyridine derivative, wherein said derivative is (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate of formula (I) or a pharmaceutically acceptable salt thereof, comprising the following step: reacting an optically active compound of Formula II ##STR00042## with a compound of Formula VII ##STR00043## or a salt thereof in the presence of a base, to obtain the compound of Formula I, ##STR00044## wherein R.sup.7 is a C.sub.1-6 alkyl, trifluoromethyl, pentafluoroethyl, heptafluoropropyl, phenyl, or a Z-substituted phenyl, in which Z is a C.sub.1-3 alkyl, halo, cyano, nitro, or trifluoromethyl, and is at position 2, 3 or 4 of the phenyl ring.

3. The method according to claim 1, wherein in the step (1), a reaction solvent is used and is one or more selected from benzene, toluene, chloroform, n-hexane, cyclohexane, dichloromethane, 1,2-dichloroethane, methyl t-butyl ether, carbon tetrachloride, ethyl acetate, propyl acetate, butyl acetate, methanol, ethanol, acetone, tetrahydrofuran, diethyl ether, acetonitrile, N,N-dimethyl formamide, or dimethyl sulfoxide; the base used is selected from triethylamine, diisopropyl ethylamine, 1,8-diazabicyclo[5,4,0]undec-7-ene, potassium carbonate, sodium carbonate, potassium bicarbonate, or sodium bicarbonate; the reaction temperature is from 20 C. to 100 C.; and the salt of the compound of Formula III is selected from hydrochloride, p-toluenesulfonate, acetate, sulfate, phosphate, trifluoromethane sulfonate, oxalate, methanesulfonate, benzenesulfonate, or hydrobromide; and wherein in the step (2), a reaction solvent is used and is one or more selected from benzene, toluene, chloroform, n-hexane, cyclohexane, dichloromethane, 1,2-dichloroethane, methyl t-butyl ether, carbon tetrachloride, ethyl acetate, propyl acetate, butyl acetate, methanol, ethanol, acetone, tetrahydrofuran, diethyl ether, acetonitrile, N,N-dimethyl formamide, or dimethyl sulfoxide; the base used is selected from triethylamine, sodium hydride, potassium hydride, 1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine, diisopropylethylamine, lithium diisopropylamide, potassium carbonate, sodium carbonate, potassium bicarbonate, sodium bicarbonate, potassium t-butoxide, or sodium t-butoxide; and the reaction temperature is from 20 C. to 100 C.

4. The method according to claim 3, wherein in the step (1), said reaction solvent is one or more selected from N,N-dimethyl formamide, tetrahydrofuran, acetonitrile, or dichloromethane; and said reaction temperature is from 10 C. to 60 C.

5. The method according to claim 3, wherein in the step (2), said reaction solvent is one or more selected from tetrahydrofuran, acetonitrile, or N,N-dimethyl formamide; and said reaction temperature is from 0 C. to 50 C.

6. The method according to claim 2, wherein a reaction solvent is used and is one or more selected from benzene, toluene, chloroform, n-hexane, cyclohexane, dichloromethane, 1,2-dichloroethane, methyl t-butyl ether, carbon tetrachloride, ethyl acetate, propyl acetate, butyl acetate, methanol, ethanol, acetone, tetrahydrofuran, diethyl ether, acetonitrile, N,N-dimethyl formamide, or dimethyl sulfoxide; the base used is selected from triethylamine, diisopropyl ethylamine, 1,8-diazabicyclo[5,4,0]undec-7-ene, potassium carbonate, sodium carbonate, potassium bicarbonate, or sodium bicarbonate; the reaction temperature is from 20 C. to 100 C.; and the salt of the compound of Formula VII is selected from hydrochloride, p-toluenesulfonate, acetate, sulfate, phosphate, trifluoromethane sulfonate, oxalate, methanesulfonate, benzenesulfonate, or hydrobromide.

7. The method according to claim 6, wherein said solvent is one or more selected from N,N-dimethyl formamide, tetrahydrofuran, acetonitrile, or dichloromethane; and the reaction temperature is from 10 C. to 60 C.

8. A method for preparing an optically active 2-hydroxytetrahydro thienopyridine derivative, wherein said derivative is (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate of formula (I) or a pharmaceutically acceptable salt thereof, comprising the following step: reacting the optically active compound of Formula IV ##STR00045## or the salt thereof with a compound of Formula V ##STR00046## or a compound of Formula VI ##STR00047## in the presence of a base, to obtain the compound of Formula I, ##STR00048##

9. The method according to claim 8, wherein a reaction solvent is used and is one or more selected from benzene, toluene, chloroform, n-hexane, cyclohexane, dichloromethane, 1,2-dichloroethane, methyl t-butyl ether, carbon tetrachloride, ethyl acetate, propyl acetate, butyl acetate, methanol, ethanol, acetone, tetrahydrofuran, diethyl ether, acetonitrile, N,N-dimethyl formamide, or dimethyl sulfoxide; the base used is selected from triethylamine, sodium hydride, potassium hydride, 1,8-diazabicyclo[5,4,0]undec-7-ene, pyridine, diisopropylethylamine, lithium diisopropylamide, potassium carbonate, sodium carbonate, potassium bicarbonate, sodium bicarbonate, potassium t-butoxide, or sodium t-butoxide; and the reaction temperature is from 20 C. to 100 C.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows .sup.1H NMR spectra of Compound I-2.

(2) FIG. 2 shows chiral HPLC analysis of Compound I-2.

(3) FIG. 3 shows chiral HPLC analysis of Compound I-2 (an enantiomer of I-2).

(4) FIG. 4 shows chiral HPLC analysis of Compound I-2 (a racemic mixture of I-2).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

(5) The contents of the present invention will be described in detail by means of examples. The examples are provided herein for illustrative purposes only, and are not intended to limit the scope of the present invention.

Example 1

(R)-Methyl o-chloromandelate

(6) ##STR00007##

(7) (R)-o-chloromandelic acid (5.6 g) was dissolved in 23.1 ml of methanol, and then a catalytic amount of concentrated sulfuric acid (0.12 ml) was added, and heated to reflux for 2 hrs. After cooling, methanol was evaporated under reduced pressure, and the residue was dissolved in dichloromethane, and washed sequentially with 10% aqueous potassium carbonate solution and water. The dichloromethane solution was dried, and evaporated to dryness, to give 5.79 g of (R)-methyl o-chloromandelate as a clear colorless oil. Yield 96%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 3.52 (d, 1H, J=4.8 Hz), 3.78 (s, 3H), 5.57 (d, 1H, J=4.5 Hz), 7.26-7.31 (m, 2H), 7.37-7.41 (m, 2H); ESI-MS m/z 222.9 [M+Na].sup.+.

Example 2

(R)-Methyl 2-(2-chlorophenyl)-2-(4-nitrophenylsulfonyloxy)-acetate (II-1)

(8) ##STR00008##

(9) (R)-Methyl o-chloromandelate (98.4 g, 0.49 mol, ee=99%) was dissolved in 500 ml of anhydrous dichloromethane, and then 91 ml of triethylamine (0.65 mol) and a catalytic amount of DMAP were added. 120 g (0.54 mol) of p-nitrophenylsulfonyl chloride was dissolved in 500 ml of anhydrous dichloromethane, which was added dropwise into the reaction solution at 0 C., and reacted for 4-5 hrs at 0 C. Water (500 ml) was added to the reaction solution and the layers were separated. The aqueous phase was extracted with dichloromethane (150 ml3), and the organic phases were combined and dried, and dichloromethane was evaporated under reduced pressure to afford 206.5 g of a crude product as a dark red oil, which was recrystallized in methanol to obtain 154.5 g of a solid product (II-1). Yield 82%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 3.57 (s, 3H), 6.39 (s, 1H), 7.21-7.39 (m, 4H), 8.07 (d, 2H, J=8.9 Hz), 8.30 (d, 2H, J=8.9 Hz); ESI-MS ink 408.0 [M+Na].sup.+.

Example 3

(2S)-Methyl 2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-2-(2-chlorophenyl)-acetate (IV-1)

(10) ##STR00009##

(11) 58.1 g (0.15 mol) of (R)-methyl 2-(2-chlorophenyl)-2-(4-nitrophenylsulfonyloxy)-acetate (II-1), 32.3 g (0.17 mol) of 5,6,7,7a-tetrahydrothieno[3,2-c]pyridin-2(4H)-one hydrochloride (III-1), and 37.8 g (0.38 mol) of potassium bicarbonate were added to 500 ml of acetonitrile. The reaction was stirred under a nitrogen atmosphere at room temperature for 26 hrs. The reaction solution was allowed to stand and the insoluble material was filtered off, to obtain a dark red mother liquor. The solvent was evaporated under reduced pressure, and 35.4 g of an oil product was obtained after purification by flash column chromatography (petroleum ether:ethyl acetate=4:1). Yield 70%. Recrystallization from ethanol afforded 18.1 g of a pure product (IV-1) as a white solid. mp: 146-148 C., ee=97.5%, [].sub.D.sup.19=+114.0 (c 0.5, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.79-1.93 (m, 1H), 2.30-2.40 (m, 1H), 2.56-2.70 (m, 1H), 3.00-3.27 (m, 2H), 3.72 (s, 3H), 3.79-3.93 (m, 1H), 4.12-4.19 (m, 1H), 4.89 (d, 1H, J=5.6 Hz), 6.00 (d, 1H, J=5.2 Hz), 7.26-7.50 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 33.9, 34.0, 49.0, 49.7, 51.1, 51.6, 52.2, 52.4, 67.3, 76.6, 77.0, 77.4, 126.6, 126.8, 127.2, 129.8, 130.1, 132.7, 134.8, 167.2, 167.4, 170.8, 198.6; ESI-MS m/z 338.1 [M+H].sup.+; HRMS Calcd for C.sub.16H.sub.17NO.sub.3SCl [M+H].sup.+ m/z 338.0618, found 338.0626.

Example 4

(S)-Methyl 2-(2-benzoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-1)

(12) ##STR00010##

(13) (2S)-Methyl 2-(2-chlorophenyl)-2-(2-oxo-5,6,7,7a-tetrahydrothieno[3,2-c]pyridinyl)acetate (IV-1) (113 mg) was dissolved in acetonitrile (10 ml), 0.10 ml of triethylamine was added, and 151 mg of benzoic anhydride was added dropwise at 0 C., and then the mixture was warmed to room temperature and reacted for 2 hrs. The reaction solution was poured into water (30 ml), the aqueous phase was extracted with ethyl acetate (50 ml3), and the organic phase was washed with saturated aqueous sodium bicarbonate solution and saturated saline, dried over anhydrous sodium sulfate, and evaporated, to obtain a crude product, which was subjected to flash column chromatography (petroleum ether:ethyl acetate=40:3), to obtain (S)-methyl 2-(2-benzoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-1) (77 mg). Yield 52%, mp: 84-86 C., ee=93.5% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 90% n-hexane/10% isopropanol/0.1% diethylamine; flow rate: 0.5 ml/min; and detection wavelength: UV 254 nm), [].sub.D.sup.20=+34.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.82-2.93 (m, 4H), 3.57-3.68 (m, 2H), 3.73 (s, 3H), 4.95 (s, 1H), 6.42 (s, 1H), 7.26-8.17 (m, 9H); .sup.13C-NMR (75 MHz CDCl.sub.3) 25.0, 48.2, 50.4, 52.2, 67.8, 112.1, 125.9, 127.2, 128.5, 128.6, 129.5, 129.8, 130.0, 130.2, 133.9, 134.7, 149.9, 163.5; ESI-MS m/z 442.1 [M+H].sup.+; HRMS Calcd for C.sub.23H.sub.21NO.sub.4SCl [M+H].sup.+ m/z 442.0891, found 442.0880.

Example 5

(S)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-2)

(14) ##STR00011##

(15) Following the method described in Example 4, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-5, 6, 7, 7a-tetrahydrothieno[3,2-c]pyridinyl)acetate (IV-1) (6.5 g) was reacted with acetic anhydride (3.6 ml), to prepare (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-2) (6.8 g). Yield 93%. Recrystallization from ethanol afforded a white solid, mp: 73-75 C., ee=98.9% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 92% n-hexane/8% tetrahydrofuran/0.1% diethylamine; flow rate: 0.5 ml/min; and detection wavelength: UV 254 nm), [].sub.D.sup.23=+45.00 (c=1.0, CH.sub.3OH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.26 (s, 3H), 2.65-2.90 (m, 4H), 3.47-3.69 (m, 2H), 3.72 (s, 3H), 4.92 (s, 1H), 6.26 (s, 1H), 7.24-7.70 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 20.2, 24.5, 47.6, 49.8, 51.6, 67.3, 111.5, 125.3, 126.6, 128.8, 128.9, 129.3, 129.4, 133.3, 134.2, 149.1, 167.2, 170.7; ESI-MS m/z 380.0 [M+H].sup.+; HRMS Calcd for C.sub.18H.sub.19NO.sub.4SCl [M+H].sup.+ m/z 380.0723, found 380.0737.

Example 6

(R)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl-acetate (I-2)

(16) ##STR00012##

(17) Following the method described in Example 4, (2R)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (prepared following Examples I-3) was reacted with acetic anhydride, to prep are (R)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-2), ee=98.2% (chiral HPLC analysis conditions were the same as those in Example 5), [].sub.D.sup.23=44.00 (c=1.0, CH.sub.3OH).

Example 7

(S)-Methyl 2-(2-propanoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-3)

(18) Following the method described in Example 4, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with propionic anhydride (0.27 ml), to prepare (S)-methyl 2-(2-propanoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-3) (267 mg). Yield 66%, ee=96.5% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20=+36.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.23 (t, 3H, J=7.4 Hz), 2.55 (q, 2H, J=7.7 Hz), 2.76-2.78 (m, 2H), 2.87-2.88 (m, 2H), 3.53 (d, 1H, J=14.2 Hz), 3.65 (d, 1H, J=13.6 Hz), 3.72 (s, 3H), 4.91 (s, 1H), 6.26 (s, 1H), 7.26-7.69 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 8.8, 21.1, 25.0, 27.4, 48.2, 50.3, 52.2, 67.8, 106.2, 111.7, 125.6, 127.2, 129.1, 129.5, 129.8, 130.0, 123.7, 149.8, 171.2; ESI-MS m/z 394.1 [M+H].sup.+; HRMS Calcd for C.sub.19H.sub.21NO.sub.4SCl [M+H].sup.+ m/z 394.0883, found 394.0880.

Example 8

(S)-methyl 2-(2-butanoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-4)

(19) ##STR00013##

(20) Following the method described in Example 4, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (86 mg) was reacted with butyric anhydride (90 l), to prepare (S)-methyl 2-(2-butanoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H-yl)-2-(2-chlorophenyl)-acetate (I-4) (51 mg). Yield 49%, ee 96.3% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20=+32.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.00 (t, 3H, J=5.2 Hz), 1.74 (q, 2H, J=5.2 Hz), 2.47-2.52 (m, 2H), 2.76-2.78 (m, 2H), 2.86-2.89 (m, 2H), 3.53 (d, 1H, J=14.3 Hz), 3.65 (d, 1H, J=14.2 Hz), 3.72 (s, 3H), 4.90 (s, 1H), 6.25 (s, 1H), 7.24-7.69 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 13.5, 18.2, 25.0, 35.8, 48.2, 50.3, 52.1, 67.9, 111.8, 125.7, 127.1, 129.2, 129.4, 129.8, 130.0, 133.8, 134.7, 149.7, 170.4, 171.2; ESI-MS m/z 408.1 [M+H].sup.+; HRMS Calcd for C.sub.20H.sub.23NO.sub.4SCl [M+H].sup.+ m/z 408.1035, found 408.1036.

Example 9

(S)-methyl 2-(2-(2-acetoxybenzoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-5)

(21) ##STR00014##

(22) 338 mg of (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) was dissolved in 15 ml of tetrahydrofuran, 0.83 ml of triethylamine was added dropwise and stirred for 10 minutes, and then 600 mg of acetylsalicylic chloride was added and stirred at room temperature for 24 hrs. The reaction solution was poured into saturated aqueous NaHCO.sub.3 solution (20 ml), extracted with ethyl acetate (50 ml3), concentrated, evaporated, and subjected to flash column chromatography (petroleum ether:ethyl acetate=5:1), to obtain (S)-methyl 2-(2-(2-acetoxybenzoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-5) (280 mg). Yield: 56%, ee=96.1% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 85% n-hexane/15% tetrahydrofuran/0.1% diethylamine; flow rate: 0.5 ml/min; and detection wavelength: UV 254 nm), [].sub.D.sup.20=+14.00 (c 0.50, CHCl.sub.3); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.34 (s, 3H), 2.79-2.81 (m, 2H), 2.90-2.93 (m, 2H), 3.54-3.66 (m, 2H), 3.72 (s, 3H), 4.93 (s, 1H), 6.37 (s, 1H), 7.14-7.36 (m, 4H), 7.39-8.15 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 20.93, 24.93, 48.04, 50.12, 52.12, 67.65, 112.61, 117.34, 118.97, 121.66, 124.09, 126.14, 127.12, 129.35, 129.45, 129.79, 129.92, 130.71, 132.12, 133.54, 134.71, 135.73, 149.32, 151.12, 161.32, 169.47, 171.17; ESI-MS m/z 500 [M+H].sup.+, 522 [M+Na].sup.+; HRMS Calcd for C.sub.25H.sub.23NO.sub.6SCl [M+H].sup.+ m/z 500.0931, found 500.0935.

Example 10

(S)-methyl 2-(2-nicotinoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-6)

(23) ##STR00015##

(24) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with nicotinoyl chloride (512 mg), to prepare (S)-methyl 2-(2-nicotinoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-6) (173 mg). Yield: 39%, mp: 92-94 C., ee=97.7% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 50% n-hexane/50% isopropanol/0.1% diethylamine; flow rate: 0.8 ml/min; and detection wavelength: UV 254 nm), [].sub.D.sup.20=+34.00 (c 0.50, MeOH); .sup.1H-NMR (500 MHz, CDCl.sub.3) 2.80-2.82 (m, 2H), 2.91-2.93 (m, 2H), 3.58 (d, 1H, J=14.3 Hz), 3.69 (d, 1H, J=14.3 Hz), 3.71 (s, 3H), 4.93 (s, 1H), 6.45 (s, 1H), 7.25-7.45 (m, 4H), 7.69 (m, 1H), 8.38-8.41 (m, 1H), 8.83-8.84 (m, 1H), 9.34 (m, 1H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 24.93, 48.05, 50.24, 52.11, 67.73, 112.43, 123.47, 124.66, 126.33, 127.11, 129.32, 129.43, 129.78, 129.89, 133.59, 134.68, 137.54, 149.18, 151.19, 154.07, 162.17, 171.16; ESI-MS m/z 443.1 [M+H].sup.+, 465.1 [M+Na].sup.+; HRMS Calcd for C.sub.22H.sub.20N.sub.2O.sub.4SCl [M+H].sup.+ m/z 443.0839, found 443.0832.

Example 11

(S)-methyl 2-(2-pivaloyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-7)

(25) Preparation Method (1)

(26) ##STR00016##

(27) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3.2-c pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with pivaloyl chloride (738 l), to prepare (S)-methyl 2-(2-pivaloyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-7) (360 mg). Yield: 85%, mp: 105-107 C., ee=99.1% (chiral HPLC analysis conditions were the same as those in Example 5), [].sub.D.sup.20=+38.00. (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.30 (s, 9H), 2.77-2.79 (m, 2H), 2.87-2.88 (m, 2H), 3.53 (d, 1H, J=14.2 Hz), 3.65 (d, 1H, J=14.2 Hz), 3.72 (s, 3H), 4.90 (s, 1H), 6.26 (s, 1H), 7.23-7.69 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 24.93, 26.95, 39.09, 48.12, 50.29, 52.09, 67.77, 111.39, 125.53, 127.10, 129.01, 129.38, 129.76, 129.92, 133.73, 134.68, 150.08, 171.20, 175.17; ESI-MS m/z 422.2 [M+H].sup.+; HRMS Calcd for C.sub.21H.sub.25NO.sub.4SCl [M+H].sup.+ m/z 422.1198, found 422.1193.

(28) Preparation Method (2)

(29) ##STR00017##

(30) 58.1 mg (0.15 mmol) of (R)-methyl 2-(2-chlorophenyl)-2-(4-nitrophenylsulfonyloxy)-acetate (II-1), 47 mg (0.17 mmol) of 4,5,6,7-tetrahydrothieno[3.2-c]pyridin-2-yl pivalate hydrochloride (VII-1) (prepared following the method described in U.S. Pat. No. 5,190,938) and 38 mg (0.38 mmol) of potassium bicarbonate were added to 5 ml of acetonitrile. The reaction was stirred under a nitrogen atmosphere at room temperature overnight. After the reaction solution was allowed to stand, the insoluble material was filtered off, and the solvent was evaporated under reduced pressure. After flash column chromatography (petroleum ether:ethyl acetate=4:1), (S)-methyl 2-(2-pivaloyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H-yl)-2-(2-chlorophenyl)-acetate (I-7) (43 mg) was obtained. Yield 69%.

Example 12

(S)-Methyl 2-(2-(2,2-dimethylbutanoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-8)

(31) ##STR00018##

(32) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7, 7a-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with 2,2-dimethylbutanoyl chloride (824 l), to prepare (S)-methyl 2-(2-(2,2-dimethylbutanoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-8) (326 mg). Yield: 75%, mp: 98-100 C., ee=99.5% (chiral HPLC analysis conditions were the same as those in Example 5), [].sub.D.sup.20=+36.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 0.88 (t, 3H, J=7.4 Hz), 1.26 (s, 6H), 1.64 (q, 2H, J=7.3 Hz), 2.76-2.78 (m, 2H), 2.87-2.88 (m, 2H), 3.53 (d, 1H, J=14.3 Hz), 3.65 (d, 1H, J=14.3 Hz), 3.71 (s, 3H), 4.90 (s, 1H), 6.25 (s, 1H), 7.22-7.69 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 9.2, 24.47, 24.99, 33.32, 43.11, 48.16, 50.31, 52.11, 67.82, 111.49, 125.59, 127.11, 129.10, 129.40, 129.78, 129.95, 133.78, 134.70, 150.03, 171.24, 174.73; ESI-MS m/z 436.2 [M+H].sup.+; HRMS Calcd for C.sub.22H.sub.27NO.sub.4SCl [M+H].sup.+, m/z 436.1352, found 436.1349.

Example 13

(S)-methyl 2-(2-cinnamoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-9)

(33) ##STR00019##

(34) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7, 7a-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with cinnamoyl chloride (1.0 g), to prepare (S)-methyl 2-(2-cinnamoyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chloro phenyl)-acetate (I-9) (162 mg). Yield: 35%, mp: 122-124 C., ee=98.7% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 90% n-hexane/10% tetrahydrofuran/0.1% diethylamine; flow rate: 0.5 ml/min; and detection wavelength: UV 254 nm), [].sub.D.sup.20=+14.00 (c 0.50, CHCl.sub.3); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.79-2.81 (m, 2H), 2.89-2.90 (m, 2H), 3.51 (d, 1H, J=18.3 Hz), 3.63 (d, 1H, J=20.3 Hz), 3.72 (s, 3H), 4.92 (s, 1H), 6.35 (s, 1H), 6.56 (d, 1H, J=15.9 Hz), 7.24-7.32 (m, 2H), 7.39-7.42 (m, 4H), 7.55-7.58 (m, 2H), 7.69-7.71 (m, 1H), 7.85 (d, 1H, J=15.9 Hz); .sup.13C-NMR (75 MHz, CDCl.sub.3) 25.01, 48.17, 50.36, 52.15, 67.83, 111.78, 116.04, 125.84, 127.15, 128.36, 128.66, 128.99, 129.44, 129.81, 130.90, 133.98, 134.72, 147.39, 163.69, 171.20; ESI-MS m/z 468.2 [M+H].sup.+, 490.2 [M+Na].sup.+; HRMS Calcd for C.sub.25H.sub.23NO.sub.4SCl [M+H].sup.+ m/z 468.1032, found 468.1036.

Example 14

(S)-methyl 2-(2-(4-nitrobenzoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-10)

(35) ##STR00020##

(36) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7, 7a-dihydrothieno[3.2-c]pyridine-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with p-nitrobenzoyl chloride (1.13 g), to prepare (S)-methyl 2-(2-(4-nitrobenzoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-10) (125 mg). Yield: 26%, mp: 100-102 C., ee=100% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 50% n-hexane/50% isopropanol/0.1% diethylamine; flow rate: 0.5 ml/min; and detection wavelength: UV 254 nm), [].sub.D.sup.20=+30.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.82-2.84 (m, 2H), 2.91-2.95 (m, 2H), 3.59 (d, 1H, J=14.3 Hz), 3.69 (d, 1H, J=14.3 Hz), 3.73 (s, 3H), 4.95 (s, 1H), 6.47 (s, 1H), 7.26-7.70 (m, 4H), 8.18 (s, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 25.14, 29.76, 48.17, 49.76, 50.38, 51.15, 51.69, 52.28, 67.37, 67.86, 112.66, 123.86, 126.68, 127.26, 129.53, 129.85, 130.00, 131.39, 133.69, 133.98, 134.84, 149.27, 151.08, 161.75, 171.35; ESI-MS m/z 487.0 [M+H].sup.+; HRMS Calcd for C.sub.23H.sub.20N.sub.2O.sub.6SCl [M+H].sup.+ m/z 487.0736, found 487.0731.

Example 15

(S)-Methyl 2-(2-(4-methoxybenzoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-11)

(37) ##STR00021##

(38) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)acetate (IV-1) (338 mg) was reacted with p-methoxybenzoyl chloride (1.02 g), to prepare (S)-methyl 2-(2-(4-methoxybenzoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-11) (142 mg). Yield: 30%, ee=96.9% (chiral HPLC analysis conditions were the same as those in Example 14), [].sub.D.sup.20=+26.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.79-2.90 (m, 4 H), 3.59-3.66 (m, 2H), 3.71 (s, 3H), 3.85 (s, 3H), 4.92 (s, 1H), 6.38 (s, 1H), 6.92-6.95 (m, 2H), 7.26-7.68 (m, 4H), 8.06-8.09 (m, 2H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 25.05, 48.15, 50.36, 52.09, 55.49, 67.85, 111.86, 113.62, 114.14, 120.66, 121.24, 125.79, 127.13, 129.43, 129.79, 129.95, 132.31, 132.77, 133.78, 134.69, 149.95, 163.17, 164.58, 171.22; ESI-MS m/z 472.2 [M+H].sup.+, 494.2 [M+Na].sup.+; HRMS Calcd for C.sub.24H.sub.23NO.sub.5SCl [M+H].sup.+ m/z 472.0993, found 472.0985.

Example 16

(S)-Methyl 2-(2-phenylacetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-12)

(39) ##STR00022##

(40) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (338 mg) was reacted with phenylacetyl chloride (796 l), to prepare (S)-methyl 2-(2-phenylacetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-12) (210 mg). Yield: 46%, ee=93.5% (chiral HPLC analysis conditions were the same as those in Example 4), [].sup.20=+14.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.75-2.77 (m, 2H), 2.86-2.88 (m, 2H), 3.52 (d, 1H, J=14.3 Hz), 3.63 (d, 1H, J=14.3 Hz), 3.71 (s, 3H), 3.82 (s, 2H), 4.89 (s, 1H), 6.26 (s, 1H), 7.23-7.41 (m, 8H), 7.64-7.68 (m, 1H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 24.98, 29.69, 40.87, 48.13, 50.29, 52.13, 67.80, 111.90, 125.81, 127.13, 127.45, 128.72, 129.14, 129.26, 129.43, 129.80, 129.92, 132.75, 133.70, 134.71, 149.62, 168.30, 171.25; ESI-MS m/z 456.2 [M+H].sup.+, 478.2 [M+Na].sup.+; HRMS Calcd for C.sub.24H.sub.23NO.sub.4SCl [M+H].sup.+ m/z 456.1041, found 456.1036.

Example 17

(S)-Methyl 2-(2-(phenoxyacetoxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-13)

(41) ##STR00023##

(42) Following the method described in Example 9, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (169 mg) was reacted with phenoxyacetyl chloride (456 mg), to prepare (S)-methyl 2-(2-(phenoxyacetoxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-13) (85 mg). Yield: 36%, mp: 104-106 C., ee=89% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20=+32.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.77-2.78 (m, 2H), 2.86-2.90 (m, 2H), 3.53 (d, 1H, J=14.3 Hz), 3.65 (d, 1H, J=14.3 Hz), 3.71 (s, 3H), 4.82 (s, 2H), 4.90 (s, 1H), 6.32 (s, 1H), 6.92-6.95 (m, 2H), 6.99-7.04 (m, 1H), 7.24-7.42 (m, 5H), 7.65-7.68 (m, 1H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 24.95, 48.07, 50.23, 52.15, 65.14, 67.72, 112.41, 114.80, 122.13, 126.14, 127.15, 129.29, 129.48, 129.65, 129.83, 129.93, 133.58, 134.73, 148.79, 157.56, 165.88, 171.17; ESI-MS m/z 472.2 [M+H].sup.+, 494.2 [M+Na].sup.+; HRMS Calcd for C.sub.24H.sub.23NO.sub.5SCl [M+H].sup.+ m/z 472.0993, found 472.0985.

Example 18

(S)-Methyl 2-(2-(ethoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-14)

(43) ##STR00024##

(44) (2S)-Methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (85 mg) was dissolved in 10 ml of anhydrous tetrahydrofuran, and 0.141 ml of triethylamine was added at room temperature and stirred for 10 min. Then, 0.167 ml of ethyl chloroformate was added dropwise at 0 C. After the addition, the reaction solution became a light yellow cloudy solution from a dark red clear solution. The ice bath was removed and the reaction was warmed to 10 C. for 1 hr. Then, 0.07 ml of triethylamine and 0.1 ml of ethyl chloroformate were additionally added in an ice bath at 0 C. The ice bath was removed and the reaction was warmed to 10 C. for 1 hr, at which the raw materials substantially disappeared. Water (20 ml) was added, the reaction solution was extracted with ethyl acetate (30 ml3) and dried over anhydrous sodium sulfate, and the solvent was evaporated to give a crude product, which was subjected to flash column chromatography (petroleum ether:ethyl acetate=40:3) to obtain (S)-methyl 2-(2-(ethoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-14) (77 mg). Yield 75%, mp: 42-44 C., ee=97.3% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20=+40.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.37 (t, 3H, J=7.2 Hz), 2.76-2.78 (m, 2H), 2.86-2.90 (m, 2H), 3.52 (d, 1H, J=14.2 Hz), 3.64 (d, 1H, J=14.3 Hz), 3.72 (s, 3H), 4.32 (q, 2H, J=7.1, 14.3 Hz), 4.90 (s, 1H), 6.30 (s, 1H), 7.24-7.68 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 14.4, 25.4, 48.4, 50.5, 52.4, 65.7, 68.1, 112.7, 126.2, 127.4, 129.7, 129.8, 130.1, 130.2, 134.0, 135.0, 152.9, 157.3, 171.5; ESI-MS m/z 410.1 [M+H].sup.+, 432.1 [M+Na].sup.+; HRMS Calcd for C.sub.19H.sub.21NO.sub.5SCl [M+H].sup.+ m/z 410.0836, found 410.0829.

Example 19

(S)-Methyl 2-(2-(isobutoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-15)

(45) ##STR00025##

(46) Following the method described in Example 18, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (85 mg) was reacted with isobutyl chloroformate (0.229 ml), to prepare (S)-methyl 2-(2-(isobutoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-15) (75 mg). Yield 69%, ee=95.5% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20=+16.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 0.98 (d, 6H, J=6.7 Hz), 2.01-2.05 (m, 1H), 2.76-2.78 (m, 2H), 2.86-2.90 (m, 2H), 3.52 (d, 1H, J=14.3 Hz), 3.64 (d, 1H, J=14.4 Hz), 3.72 (s, 3H), 4.25 (d, 2H, J=6.6 Hz), 4.90 (s, 1H), 6.29 (s, 1H), 7.26-7.42 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 19.1, 25.4, 28.0, 48.4, 50.5, 52.4, 68.1, 75.6, 112.6, 126.1, 127.4, 129.7, 129.8, 130.1, 130.2, 134.0, 135.0, 150.8, 153.0, 171.5; ESI-MS m/z 460.3 [M+Na].sup.+; HRMS Calcd for C.sub.21H.sub.25NO.sub.5SCl [M+H].sup.+ m/z 438.1150, found 438.1142.

Example 20

(S)-Methyl 2-(2-(isopropoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-16)

(47) ##STR00026##

(48) Following the method described in Example 18, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7, 7a-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (85 mg) was reacted with isopropyl chloroformate (0.23 ml), to prepare (S)-methyl 2-(2-(isobutoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-15) (100 mg). Yield: 94%, ee=97.5% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20=+34.00 (c 0.50, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.36 (d, 6H, J=6.2 Hz), 2.77-2.78 (m, 2H), 2.86-2.89 (m, 2H), 3.52 (d, 1H, J=14.2 Hz), 3.64 (d, 1H, J=14.3 Hz), 3.72 (s, 3H), 4.90 (s, 1H), 4.93-5.01 (m, 1H), 6.30 (s, 1H), 7.24-7.68 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 21.6, 25.4, 48.1, 50.3, 52.1, 67.8, 73.9, 112.2, 125.7, 127.1, 129.4, 129.8, 129.9, 130.3, 133.7, 134.7, 150.5, 152.0, 171.2; ESI-MS m/z 424.1 [M+H].sup.+; HRMS Calcd for C.sub.20H.sub.23NO.sub.5SCl [M+H].sup.+ m/z 424.0989, found 424.0985.

Example 21

(S)-Methyl 2-(2-(benzyloxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-17)

(49) ##STR00027##

(50) Following the method described in Example 18, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (85 mg) was reacted with benzyl chloroformate (0.25 ml), to prepare (S)-methyl 2-(2-(benzyloxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-17) (97 mg). Yield 82%, ee=93.7% (chiral HPLC analysis conditions were the same as those in example 4), [].sub.D.sup.20=+12.00 (c 0.50, CHCl.sub.3); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.76-2.78 (m, 2H), 2.86-2.90 (m, 2H), 3.52 (d, 1H, J=14.4 Hz), 3.63 (d, 1H, J=14.4 Hz), 3.72 (s, 3H), 4.90 (s, 1H), 5.26 (s, 2H), 6.30 (s, 1H), 7.24-7.42 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 25.1, 48.1, 50.3, 52.1, 67.8, 70.9, 112.5, 126.0, 127.0, 127.1, 128.6, 128.7, 128.9, 129.4, 129.5, 129.8, 129.9, 133.7, 134.3, 134.7, 150.4, 152.6, 171.2; ESI-MS m/z 472.1 [M+H].sup.+, 494.1 [M+Na].sup.+; HRMS Calcd for C.sub.24H.sub.23NO.sub.5SCl [M+H].sup.+ m/z 472.0996, found 472.0985.

Example 22

(S)-Methyl 2-(2-(N,N-dimethylcarbamoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-18)

(51) ##STR00028##

(52) In an ice bath, sodium hydride (20 mg, 60%) were added to 6 ml of anhydrous N, N-dimethyl formamide, to which 169 mg of (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridine-5(2H,4H,6H)-yl)-acetate (IV-1) was then added. The reaction was naturally warmed to room temperature, stirred for 1 hr, and then cooled to 0 C. 200 l of N,N-dimethylcarbamoyl chloride was slowly added dropwise. The reaction was naturally warmed to room temperature, and stirred for 3 hrs, and at this time, TLC showed that the raw materials substantially disappeared. The reaction solution was poured into water (30 ml), the aqueous phase was extracted with ethyl acetate (50 ml3), and the organic phase was dried to obtain a crude product, which was subjected to flash column chromatography (petroleum ether:ethyl acetate=40:3), to obtain (S)-methyl 2-(2-(N,N-dimethylcarbamoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-18) (95 mg). Yield: 45%, mp 96-98 C., ee=93.5% (chiral HPLC analysis conditions were the same as those in Example 10), [].sub.D.sup.20=+34 (c 0.50, MeOH); .sup.1H-NMR (500 MHz, CDCl.sub.3) 2.73-2.76 (m, 2H), 2.85-2.89 (m, 2H), 2.99 (s, 3H), 3.03 (s, 3H), 3.52 (d, 1H, J=14.3 Hz), 3.63 (d, 1H, J=14.3 Hz), 3.72 (s, 3H), 4.91 (s, 1H), 6.19 (s, 1H), 7.26-7.69 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 25.03, 36.37, 36.88, 48.18, 50.40, 52.12, 67.87, 111.48, 125.38, 127.12, 129.09, 129.37, 129.76, 129.95, 130.56, 133.80, 134.34, 134.68, 151.04, 171.27; ESI-MS m/z 409.2 [M+H].sup.+; HRMS Calcd for C.sub.19H.sub.22N.sub.2O.sub.4SCl [M+H].sup.+ m/z 409.0992, found 409.0989.

Example 23

(S)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-fluorophenyl)-acetate (I-19)

(53) ##STR00029##

(54) Following the method described in Example 4, (2S)-methyl 2-(2-fluorophenyl)-2-(2-oxo-7,7a-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-2) (100 mg) was reacted with acetic anhydride (63 l), to prepare (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-fluorophenyl)-acetate (I-19) (98.0 mg). Yield 86%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.26 (s, 3H), 2.47-2.97 (m, 4H), 3.64 (s, 2H), 3.74 (s, 3H), 4.80 (s, 1H), 6.27 (s, 1H), 7.07-7.59 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 20.7, 24.9, 48.0, 50.0, 52.2, 64.3, 111.9, 115.5, 115.8, 124.4, 124.5, 125.6, 129.0, 130.0, 130.1, 130.2, 130.3, 149.6, 159.3, 162.6, 167.7, 171.0; ESI-MS m/z 364.1 [M+H].sup.+; HRMS Calcd for C.sub.18H.sub.19NO.sub.4SF [M+H].sup.+ m/z 364.1019, found 364.1029.

Example 24

(S)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(4-chlorophenyl)-acetate (I-20)

(55) ##STR00030##

(56) Following the method described in Example 4, (2S)-methyl 2-(4-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-3) (100 mg) was reacted with acetic anhydride (63 l), to prepare (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(4-chlorophenyl-acetate (I-20) (110 mg). Yield 97%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.27 (s, 3H), 2.65-2.78 (m, 4H), 3.53 (s, 2H), 3.73 (s, 3H), 4.30 (s, 1H), 6.25 (s, 1H), 7.33-7.46 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 20.7, 24.7, 48.1, 50.4, 52.2, 71.9, 111.9, 125.7, 128.9, 130.1, 134.3, 134.4, 149.6, 167.7, 171.4; ESI-MS m/z 380.1 [M+H].sup.+; HRMS Calcd for C.sub.18H.sub.19NO.sub.4SCl [M+H].sup.+ m/z 380.0723, found 380.0735.

Example 25

(S)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-phenyl-acetate (I-21)

(57) ##STR00031##

(58) Following the method described in Example 4, (2S)-methyl 2-phenyl-2-(2-oxo-7,7-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-4) (100 mg) was reacted with acetic anhydride (63 l), to prepare (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-phenyl-acetate (I-21) (108 mg). Yield 93%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.26 (s, 3H), 2.47-2.91 (m, 4H), 3.54 (s, 2H), 3.72 (s, 3H), 4.32 (s, 1H), 6.25 (s, 1H), 7.25-7.69 (m, 5H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 20.7, 24.4, 48.1, 50.3, 52.2, 72.5, 111.9, 128.7, 128.9, 167.7; ESI-MS m/z 346.1 [M+H].sup.+; HRMS Calcd for C.sub.18H.sub.20NO.sub.4S [M+H].sup.+ m/z 346.1113, found 346.1125.

Example 26

(S)-Methyl 2-(2-(pyrrolidine-1-carbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-22)

(59) ##STR00032##

(60) Following the method described in Example 22, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (84 mg) was reacted with pyrrolidine-1-carbonyl chloride (70 l), to prepare (S)-methyl 2-(2-(pyrrolidine-1-carbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-22) (50 mg). Yield: 46%, ee=80.1% (chiral HPLC analysis conditions: Chiralpak IC 4.6 mm250 mm; column temperature: 25 C.; mobile phase: 50% n-hexane/50% isopropanol/0.1% diethylamine; flow rate: 0.5 ml/min; detection wavelength: UV 254 rnn), [].sub.D.sup.22=+19.0 (c 1.0, MeOH); .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.87-2.04 (m, 4H), 2.62-2.75 (m, 2H), 2.77-2.89 (m, 2H), 3.44-3.54 (m, 5H), 3.61-3.76 (m, 4H), 4.89 (s, 1H), 6.20 (s, 1H), 7.22-7.40 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 24.9, 25.1, 25.7, 29.7, 46.3, 46.6, 48.2, 50.4, 52.1, 67.9, 111.5, 125.2, 127.1, 128.6, 129.1, 129.3, 129.7, 130.0, 133.9, 134.7, 151.0, 151.8, 171.3; ESI-MS m/z 435.1 [M+H].sup.+; HRMS Calcd for C.sub.2IH.sub.24N.sub.2O.sub.4SCl [M+H].sup.+ m/z 435.1145, found 435.1148.

Example 27

(S)-Methyl 2-(2-(methoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-23)

(61) ##STR00033##

(62) Following the method described in Example 18, (2S)-methyl 2-(2-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (108.5 mg) was reacted with 0.1 ml of methyl chloroformate, to prepare (S)-methyl 2-(2-(methoxycarbonyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-23) (86 mg). Yield: 68%, ee=97.0% (chiral HPLC analysis conditions were the same as those in Example 4), [].sub.D.sup.20+24.00 (c 0.50, CHCl.sub.3); .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.75-2.78 (m, 2H), 2.83-2.90 (m, 2H), 3.47-3.68 (m, 2H), 3.72 (s, 3H), 3.90 (s, 3H), 4.90 (s, 1H), 6.29 (s, 1H), 7.23-7.68 (m, 4H); .sup.13C-NMR (75 MHz, CDCl.sub.3) 25.1, 48.1, 50.3, 52.1, 55.8, 67.8, 112.6, 126.0, 127.1, 129.4, 129.6, 129.8, 129.9, 133.7, 134.7, 150.4, 153.3, 171.2; ESI-MS m/z 396.1 [M+H].sup.+. HRMS Calcd for C.sub.18H.sub.19NO.sub.5SCl [M+H].sup.+ m/z 396.0672, found 396.0675.

Example 28

(R,S)-Methyl 2-(2-(N,N-dimethylcarbamoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-18)

(63) ##STR00034##

(64) Following the method described in Example 22, methyl 2-(R,S)-2-(2-chlorophenyl)-2-(2-oxo-7,7-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-5) (169 mg) was reacted with N,N-dimethylcarbamoyl chloride (200 l), to prepare (R,S)-methyl 2-(2-(N,N-dimethylcarbamoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-18) (185 mg). Yield: 87%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.73-2.76 (m, 2H), 2.85-2.90 (m, 2H), 2.98 (s, 3H), 3.02 (s, 3H), 3.52 (d, 1H, J=14.3 Hz), 3.62 (d, 1H, J=14.3 Hz), 3.72 (s, 3H), 4.91 (s, 1H), 6.19 (s, 1H), 7.26-7.69 (m, 4H); ESI-MS m/z 409 [M+H].sup.+.

Example 29

(R,S)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-2)

(65) ##STR00035##

(66) Following the method described in Example 4, methyl 2-(R,S)-2-(2-chlorophenyl-7,7-dihydrothieno[3.2-c]pyridin-5(2H,4H,6H)-yl)-acetate (IV-1) (650 mg) was reacted with acetic anhydride (1 ml), to prepare (R,S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-2) (670 mg). Yield 92%. .sup.1H-NMR (300 MHz, CDCl.sub.3) 2.26 (s, 3H), 2.76 (d, 2H, J=5.4 Hz), 2.90 (d, 2H, J=5.0 Hz), 3.55 (d, 1H, J=14.2 Hz), 3.64 (d, 1H, J=14.2 Hz), 3.72 (s, 3H), 4.90 (s, 1H), 6.26 (s, 1H), 7.25-7.68 (m, 4H); ESI-MS m/z 380.0 [M+H].sup.+.

Example 30

(S)-Methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate Hydrochloride (I-2 Hydrochloride)

(67) 103 mg of (S)-methyl 2-(2-acetoxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-2) was dissolved in 6 ml of diethyl ether, and stirred in an ice-salt bath at 10 C. A hydrogen chloride saturated ethanol solution (0.2 ml) was added slowly dropwise, till the system reached about pH 2, at which a white solid was precipitated immediately. It was stirred for 5 min, allowed to stand, quickly filtered under a nitrogen atmosphere, and washed with a suitable amount of diethyl ether. The resulting solid was dried in vacuum to obtain the I-2 hydrochloride (101 mg, white solid). Yield: 90%, and melting range: 100-120 C.

Example 31

(S)-methyl 2-(2-pivaloyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate Hydrochloride (I-7 Hydrochloride)

(68) 65 mg of (S)-methyl 2-(2-pivaloyloxy-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-7) was dissolved in 4 ml of diethyl ether, and stirred in an ice-salt bath at 10 C. A hydrogen chloride saturated ethanol solution (0.2 ml) was added slowly dropwise, till the system reached about pH 2, at which a white solid was precipitated immediately. It was stirred for 5 min, allowed to stand, quickly filtered under a nitrogen atmosphere, and washed with a suitable amount of diethyl ether. The resulting solid was dried in vacuum to obtain the I-7 hydrochloride (65 mg, granular solid). Yield: 92%, melting point: 135-137 C.; .sup.1H-NMR (300 MHz, CDCl.sub.3) 1.33 (s, 9H), 3.08 (s, 2H), 3.44-3.51 (m, 2H), 3.82 (s, 4H), 4.34 (s, 1H), 5.59 (s, 1H), 6.39 (s, 1H), 7.44-7.50 (m, 3H), 8.36 (d, 1H, J=7.2 Hz).

Example 32

(S)-methyl 2-(2-(2,2-dimethylbutanoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate Hydrochloride (I-8 Hydrochloride)

(69) 63 mg of (S)-methyl 2-(2-(2,2-dimethylbutanoyloxy)-6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)-2-(2-chlorophenyl)-acetate (I-8) was dissolved in 4 ml of diethyl ether, and stirred in an ice-salt bath at 10 C. A hydrogen chloride saturated ethanol solution (0.2 ml) was added slowly dropwise, till the system reached about pH 2, at which a white solid was precipitated immediately. It was stirred for 5 min, then allowed to stand, quickly filtered under a nitrogen atmosphere, and washed with a suitable amount of diethyl ether. The resulting solid was dried in vacuum to obtain the I-8 hydrochloride (60 mg, white granular solid). Yield: 88%, melting point: 133-135 C.; .sup.1H-NMR (300 MHz, CDCl.sub.3) 0.91 (t, 3H, J=7.6 Hz), 1.29 (s, 6H), 1.69 (q, 2H, J=7.4 Hz), 3.08 (s, 1H), 3.48 (m, 2H), 3.83 (s, 4H), 4.35 (s, 1H), 5.59 (s, 1H), 6.37 (s, 1H), 7.44-7.50 (m, 3H), 8.36 (d, 1H, J=6.84 Hz).

Example 33

(70) Anti-Platelet Aggregation Activity Test

(71) Agents and preparations: clopidogrel sulfate was used as a positive control. The positive control and the test compounds (prepared in the above examples) were formulated in 0.5% CMC-Na (carboxymethylcellulose sodium) into suspensions for administration to animals.

(72) Animals: male SD rats, weighed about 250 g, and supplied by Shanghai Super-B&K Laboratory Animal Corp. Ltd. Animal certificate No.: 2008001605451, and license No.: SCXK (Hu); SCXK (Hu) 2008-0016.

(73) Instruments: a centrifuge (80-2 bench top low-speed centrifuge), an automatic platelet aggregation tester (STELLEXLG-PAPER-I platelet coagulation/aggregation analyzer), and others.

(74) Methods: the pharmacological activity test of the compounds of the present invention for platelet aggregation was performed following Born's turbidimetric method (Nature, 1962, 194(4832): 927). An agglomeration-promoting substance, adenosine diphosphate (ADP), was added to a platelet rich plasma (PRP), and stirred to allow platelets to aggregate. The aggregation of platelets resulted in the change of optical density, which could be detected by a spectrometer. This experiment can be used to evaluate the platelet aggregation effect induced by the test compounds administrated in-vivo or in-vitro.

(75) Anti-platelet aggregation activity test: the male SD rates weighed about 250 g were given with clopidogrel sulfate and the test agents (homogeneous suspensions in 0.5% CMC-Na at a concentration of 1 mg/ml) by oral gavage at a dose of 10 mg/kg or 3 mg/kg, and the blank control was given with the same volume of 0.5% CMC-Na by oral gavage. After 2 hrs, blood samples were collected from eye sockets of the rats using 3.8% sodium citrate as anticoagulant at a ratio of whole blood to anticoagulant of 9:1, and centrifuged at 1000 rpm for 7 min to prepare a platelet rich plasma (PRP). The PRP was adjusted with a platelet poor plasma (PPP) to maintain the platelet counts at 210.sup.6/ml. The PRP was taken into a test cup, and incubated at 37 C. for 10 min. Light transmission was adjusted to 0% with PRP and to 100% with PPP for each measurement. By using ADP (with a final concentration of 5 M) as an inducer, the platelet aggregation percentages were measured through the turbidimetric method using a platelet aggregation tester, and statistically compared by t-test. The platelet aggregation inhibition was calculated by the equation: platelet aggregation inhibition (%)=[1(aggregation percent in dosed tube/aggregation percent in control tube)]100%.

(76) Results: the platelet aggregation inhibition after oral administration of the test compounds to the rats is determined through the turbidimetric method, and some test results are shown in Table 1. The results show that most of the test compounds exhibit a more potent anti-platelet aggregation activity than clopidogrel, and the compounds of (S)-configuration (e.g. I-2, Example 5) exhibit a more potent platelet aggregation inhibition than the corresponding enantiomers of (R)-configuration (e.g. I-2, Example 6) and racemic mixtures (e.g. I-2, Example 29).

(77) TABLE-US-00001 TABLE 1 Platelet aggregation inhibition after oral administration of test compounds to rats Aggregation Dosage Animals inhibition Test compounds (mg/kg) (n) (%) Blank control 6 Clopidogrel sulfate 10 6 79.6 3 6 15.4 I-1 (Example 4) 3 6 41.1 I-2 (Example 5) 10 6 83.5 3 6 69.9 I-2 (Example 6) 10 6 55.3 3 6 27.1 I-2 (Example 29) 10 6 78.5 3 6 43.92 I-3 (Example 7) 3 6 38.4 I-4 (Example 8) 3 6 46.2 I-5 (Example 9) 3 6 55.1 I-6 (Example 10) 3 6 31.3 I-7 (Example 11) 3 6 41.9 I-8 (Example 12) 3 6 35.7 I-9 (Example 13) 3 6 30.3 I-10 (Example 14) 3 6 36.8 I-11 (Example 15) 3 6 13.6 I-12 (Example 16) 3 6 18.3 I-13 (Example 17) 3 6 10.1 I-14 (Example 18) 3 6 62.9 I-15 (Example 19) 3 6 45.3 I-16 (Example 20) 3 6 52.7 I-18 (Example 22) 3 6 48.2 I-19 (Example 23) 3 6 63.5 I-20 (Example 24) 3 6 17.6 I-21 (Example 25) 3 6 9.1 I-22 (Example 26) 3 6 20.3 I-18 (Example 28) 3 6 29.8

Example 34

(78) Researches on Pharmacokinetics and Bioavailability of Compound I-2 in Rats

(79) Research background: it is reported (Thromb Haemost, 2000, 84: 891; Drug Metab Rev 2005, 37 (Suppl 2): 99) that oxidative metabolism of clopidogrel in vivo by the liver P450 enzyme system first produces the metabolic intermediate (2S)-methyl 2-(2-oxo-7,7a-dihydrothieno[3,2-c]pyridin-5(2H,4H,6H)-yl)-2-(2-chlorophenyl)-acetate (thiolactone IV-1, see Example 3 for preparation thereof), and then Compound IV-1 is further rapidly metabolized into the pharmacologically active metabolite. Therefore, Compound IV-1 is useful as an indicator for the production of the active metabolite of clopidogrel (Drug Metab Disp 2002, 30: 1288).

(80) Experimental purposes: (1) the plasma concentration-time curves of Compound I-2, clopidogrel, and metabolite IV-1 thereof after the rats are respectively given with Compound I-2 and clopidogrel by oral gavage are investigated, so as to estimate corresponding pharmacokinetic parameters, evaluate the pharmacokinetic characteristics of Compound I-2 and clopidogrel in rats, determine the conversion of Compound I-2 into the metabolite IV-1 and the conversion degree, and compare the respective conversion degrees of Compound I-2 and clopidogrel to the metabolite IV-1; and (2) the plasma concentration-time curves of Compound IV-1 after administration to the rats via intravenous injection are investigated, so as to estimate corresponding pharmacokinetic parameters, and evaluate the absolute bioavailability of the metabolite IV-1 produced through respective metabolism of Compound I-2 and clopidogrel in the rats.

(81) Methods: male SD rats weighed 210-230 g were divided into 3 groups at random, Compound I-2 group, clopidogrel sulfate group, and Compound IV-1 group. The rats were fasted overnight, but allowed free access to water. After 10 h, (1) the Compound I-2 group was administrated with Compound I-2 by oral gavage in a volume of 8 ml/kg at a concentration of 1.14 mg/ml at a dose of 24 mol/kg, and blood samples were collected from retro-orbital plexus in rats before dosing (0 h) and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hrs post-dose; (2) the clopidogrel sulfate group was administrated with clopidogrel sulfate by oral gavage in a volume of 8 ml/kg at a concentration of 1.26 mg/ml at a dose of 24 mol/kg, and blood samples were collected from retro-orbital plexus in rats before dosing (0 h) and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hrs post-dose; and (3) the Compound IV-1 group was administrated with Compound IV-1 via intravenous injection in a volume of 5 ml/kg at a concentration of 0.54 mg/ml at a dose of 8 mol/kg, and blood samples were collected from retro-orbital plexus in rats before dosing (0 h) and at 0.083, 0.167, 0.5, 1, 2, 4, 6, 8, and 24 hrs post-dose. Plasmas were isolated after treatment with an anticoagulant and a stabilizer, the plasma samples were processed, and the concentrations of Compound I-2, clopidogrel, and Compound IV-1 in plasmas were measured by LC-MS/MS. Chromatography and MS conditions are shown in Tables 2 and 3.

(82) TABLE-US-00002 TABLE 2 Chromatography conditions for samples and internal standard (diazepam) Chromatography ACE 3.0*50 mm, 5 m, C18, P/NO: ACE-121-0503 column Phase A: Phase B: mobile Time water + 0.1% acetonitrile + 0.1% phase (min) formic acid formic acid 0-0.5 75% 25% 0.5-1.5 75%.fwdarw.5% 25%.fwdarw.95% 1.5-3.0 5% 95% 3.0-3.2 5%.fwdarw.75% 95%.fwdarw.25% 3.2-4.0 75% 25% Flow rate 0.5 ml/min Injector 10 C. Column 35 C. Injection Total valve time Position 2.0 B 3.2 A Retention Compound I-2: 2.66 min; Clopidogrel: time 2.70 min; Compound IV-1: 2.60 min; (min) and diazepam: 2.50 min

(83) TABLE-US-00003 TABLE 3 MS conditions for samples and internal standard (diazepam) Compound Sample IV-1 Clopidogrel Compound Diazepam Ion source ESI Polarity Positive Scan mode Q1, Q3, MRM Nebulizer 40 psi gas Heater gas 40 psi Curtain gas 20 psi Spray voltage 5000 V.sup. Declustering 56 V 53 V 60 v 105 v Collision gas 36 ev 23 ev 23 ev 42 ev Temperature 500 Ion pair m/z 338.2/154.9 322.0/212.0 380.0/212.0 285.3/154.0

(84) Results: the plasma concentration-time data after administration of the test agents to the SD rats are shown in Tables 4, 5, and 6 (note: NA is not available). The results show that (1) after 3 SD rats are dosed with 24 mol/kg Compound I-2 by oral gavage, the Compound IV-1 generated by metabolism has a elimination half-life of t.sub.1/2=2.191.68 h, an area AUC.sub.0-t under the plasma concentration-time curve of 197124 g.Math.h/L, an area AUC.sub.0-, under the plasma concentration-time curve of 211119 g.Math.h/L, a peak time of T.sub.max=1.170.764 h, a peak concentration of C.sub.max=67.242.3 g/L, and an absolute bioavailability of 24.6%; (2) after 3 SD rats are dosed with 24 mol/kg clopidogrel sulfate by oral gavage, the Compound IV-1 generated by metabolism has an elimination half-life of =2.480.466 h, an area AUC.sub.0-t under the plasma concentration-time curve of 29.011.5 g.Math.h/L, an area AUC.sub.0-, under the plasma concentration-time curve of 32.210.9 g.Math.h/L, a peak time of T.sub.max=0.5830.382 h, a peak concentration of C.sub.max=6.933.36 g/L, and an absolute bioavailability of 3.63%; and (3) after 3 SD rats are dosed with 8 mol/kg Compound IV-1 via intravenous injection, the Compound IV-1 has an elimination half-life of t.sub.1/2=1.060.364 h, an area AUC.sub.0-t under the plasma concentration-time curve of 26637.6 g.Math.h/L, an area AUC.sub.0- under the plasma concentration-time curve of 26838.3 g.Math.h/L, a peak time of T.sub.max=0.0830 h, and a peak concentration of C.sub.max=671128 g/L.

(85) TABLE-US-00004 TABLE 4 Concentration-time data (ng/ml) of Compound IV-1 in plasma after administration of Compound I-2 to SD rats by oral gavage Animal Time (h) No. 0 0.25 0.5 1 2 4 6 8 24 1# 0 35.5 53.0 65.4 115 32.4 8.59 5.74 ND 10# NA 19.4 29.2 34.7 19.6 11.8 4.25 1.56 ND 11# NA 22.5 51.9 37.7 16.6 12.6 18.9 6.12 ND Mean 25.8 44.7 45.9 50.4 18.9 10.6 4.47 SD 8.54 13.4 16.9 56.0 11.7 7.53 2.53

(86) TABLE-US-00005 TABLE 5 Concentration-time data (ng/ml) of Compound IV-1 in plasma after administration of clopidogrel sulfate to SD rats by oral gavage Animal Time (h) No. 0 0.25 0.5 1 2 4 6 8 24 5# 0 8.99 10.3 10.8 7.57 4.70 3.15 0.895 ND 12# NA 4.73 4.46 4.38 4.27 2.48 1.35 0.887 ND 13# NA 4.18 5.27 5.09 3.21 2.69 2.87 0.924 ND Mean 5.97 6.68 6.76 5.02 3.29 2.46 0.902 SD 2.63 3.16 3.52 2.27 1.23 0.969 0.0195

(87) TABLE-US-00006 TABLE 6 Concentration-time data (ng/ml) of Compound IV-1 in plasma after intravenous injection of Compound IV-1 to SD rats Animal Time (h) No. 0 0.083 0.167 0.5 1 2 4 6 8 24 7# 0 696 317 108 51.0 10.9 3.94 1.29 ND ND 8# 0 785 417 129 49.3 9.06 2.87 0.977 ND ND 9# 0 533 311 107 37.3 9.74 2.02 1.06 ND ND Mean 0 671 348 115 45.9 9.90 2.94 1.11 SD 0 128 59.5 12.4 7.47 0.930 0.962 0.162

(88) The estimated pharmacokinetic parameters after the SD rats are dosed with the test agents are shown in Tables 7, 8, and 9. (Note: in Tables 7, 8, and 9, it is assumed that Compound I-2 or clopidogrel can be totally converted into Compound IV-1 in the calculation of clearance CL.sub.tot and apparent volume of distribution V.sub.z of Compound IV-1, and thus the dosage in the calculation formula is that of Compound I-2 or clopidogrel equivalent to Compound IV-1)

(89) TABLE-US-00007 TABLE 7 Pharmacokinetic parameters of metabolite (Compound IV-1) after administration of Compound I-2 to SD rats by oral gavage Equivalent of C.sub.max T.sub.max t.sub.1/2 CL.sub.tot V.sub.2 AUC.sub.0-t AUC.sub.0- 8.1 mg/kg (g/L) (h) (h) (L/h/kg) (L/kg) (g .Math. h/L) (g .Math. h/L) 1# 115 2.00 1.07 23.7 36.6 338 342 10# 34.7 1.00 1.37 75.0 148 105 108 11# 51.9 0.500 4.12 44.0 261 147 184 Mean 67.2 1.17 2.19 47.6 149 197 211 SD 42.3 0.764 1.68 25.8 112 124 119

(90) TABLE-US-00008 TABLE 8 Pharmacokinetic parameters of metabolite (Compound IV-1) after administration of clopidogrel sulfate to SD rats by oral gavage Equivalent of C.sub.max T.sub.max t.sub.1/2 CL.sub.tot V.sub.2 AUC.sub.0-t AUC.sub.0- 8.1 mg/kg (g/L) (h) (h) (L/h/kg) (L/kg) (g .Math. h/L) (g .Math. h/L) 5# 10.8 1.00 1.95 181 510 42.2 44.7 12# 4.73 0.250 2.65 331 1267 21.1 24.5 13# 5.27 0.500 2.83 294 1199 23.7 27.6 Mean 6.93 0.583 2.48 269 992 29.0 32.2 SD 3.36 0.382 0.466 77.9 419 11.5 10.9

(91) TABLE-US-00009 TABLE 9 Pharmacokinetic parameters of Compound IV-1 after intravenous injection of Compound IV-1 to SD rats Compound C.sub.max T.sub.max t.sub.1/2 CL.sub.tot V.sub.2 AUC.sub.0-t AUC.sub.0- 2.7 mg/kg (g/L) (h) (h) (L/h/kg (L/kg) (g .Math. h/L (g .Math. h/L 7# 696 0.0830 1.30 10.0 18.8 266 269 8# 785 0.0830 1.25 8.84 15.9 304 306 9# 533 0.0830 0.643 11.8 10.9 229 229 Mean 671 0.0830 1.06 10.2 15.2 266 268 SD 128 0 0.364 1.49 3.98 37.6 38.3

(92) Conclusions: (1) after being dosed to the rats by oral gavage, Compound I-2 can be converted into the main intermediate metabolite (Compound IV-1) of clopidogrel; and (2) by oral gavage, the degree of conversion of Compound I-2 into the metabolite IV-1 is more than 5 times higher than that of clopidogrel sulfate, and the absolute bioavailability of the metabolite IV-1 produced by metabolism of Compound I-2 is also more than 5 times higher than that of the metabolite IV-1 produced by metabolism of clopidogrel sulfate.

(93) The above research results suggest that the bioavailability of the intermediate metabolite of Compound I-2 is significantly higher than that of clopidogrel, such that the production of the active metabolite thereof is also significantly higher than that of clopidogrel, and thus it is expected that the risk of side effects such as bleeding of the anti-platelet aggregation agent can be reduced while rapid onset of action and high efficacy are achieved, by significantly lowering the dosage of the agent.

Example 35

(94) Tablet

(95) Compound I-2 (50 g) prepared in Example 5, hydroxypropyl methylcellulose E (150 g), starch (200 g), a suitable amount of Povidone K30, and magnesium stearate (1 g) were mixed, granulated, and tableted.