Tetrahydrothiopyranopyrimidine derivatives as anti-HIV agent

Abstract

The invention relates the tetrahydrothiopyranopyrimidine derivatives, the preparation, and the use thereof. The invention contained tetrahydrothiopyranopyrimidine derivatives, and pharmaceutical acceptable salt and prodrug with the formula I or II. Also described here are preparation of tetrahydrothiopyranopyrimidine derivatives, pharmaceutical compositions comprising these compounds as therapy and prevention for HIV.

Claims

1. A compound of tetrahydrothiopyranopyrimidine, or pharmaceutically acceptable salt thereof, the compound having a general formula I or II shown as following: ##STR00041## wherein A is S(O).sub.n or CH.sub.2; B is S(O).sub.n or CH.sub.2, while either A or B is S(O).sub.n in the compound; n=0, 1 or 2; X is O or NH; R.sup.1, R.sup.2 and R.sup.3 are independently selected from a group consisting of H, halo, CN, CF.sub.3, NH.sub.2, OH, C.sub.1-6alkyl, C.sub.1-6alkyloxy, C.sub.2-6alkenyl, trifluoromethyl, amino, hydroxy, cyanovinyl, cyanoethyl and cyclopropyl; Ar is selected from a group consisting of an optionally substituted benzyl ring, an optionally substituted benzene ring, an optionally substituted naphthalene ring, a substituted 6-membered heterocyclic ring, a substituted 5-membered heterocyclic ring, a substituted 6-membered and 5-membered heterocyclic ring, a substituted 6- and 6-membered heterocyclic ring, a substituted five-membered and five-membered heterocyclic ring, a substituted benzo five-membered heterocyclic ring and a substituted benzo-six-membered heterocyclic ring.

2. The compound of claim 1, wherein R.sup.1 and R.sup.3 are CH.sub.3, R.sup.2 is selected from a group consisting of CN, Me and CHCHCN.

3. The compound of claim 1, wherein the Ar has a general formula (a) or (b) shown as following: ##STR00042## wherein R.sup.4 is selected from a group consisting of H, CN, Me, COR.sup.5, COOR.sup.5, CONH.sub.2, CONHR.sup.5, SO.sub.2R.sup.5, SO.sub.2NH.sub.2, SO.sub.2NHR.sup.5, NO.sub.2, NH.sub.2, NHR.sup.5, NHCOR.sup.5 and NHSO.sub.2R.sup.5; R.sup.5 is selected from a group consisting of C.sub.1-10alkyl, C.sub.1-10cyclicalkyl, C.sub.1-10haloalkyl, C.sub.1-10alkenyl and C.sub.1-10aromatic alkyl.

4. The compound of claim 3, wherein the R.sup.5 is Me or CH.sub.3CO.

5. The compound of claim 3, wherein the compound have a formula (IA-1), (IA-2), (IB-1), (IB-2), (IIA-1), (IIA-2), (IIB-1), (IIB-2), (IIC-1), (IIC-2), (IID-1) or (IID-2) shown as following: ##STR00043## ##STR00044## ##STR00045## wherein, R.sup.1, R.sup.2, R.sup.3, R.sup.4 are independently selected from a group consisting of H, halo, CN, CF.sub.3, NH.sub.2, OH, C.sub.1-6alkyl, C.sub.1-6 alkyloxy, C.sub.2-6alkenyl, trifluoromethyl, amino, hydroxy, cyanovinyl, cyanoethyl and cyclopropyl.

6. The compound of claim 5, the compound is selected from a group consisting of ##STR00046## ##STR00047## ##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052##

7. The compound of claim 1, wherein the compound is prepared by the following steps: methyl mercaptoacetate (A) reacting with methyl 4-chlorobutanoate (B) to give methyl 4-((2-methoxy-2-oxoethyl) thio) butyrate (C) in sodium methoxide; the intermediate C being converted to oxotetrahydropyranyl ester (D) by Dieckmann condensation under sodium methoxide; the intermediate D being cyclized with methyl isothiourea into 2-methylmercaptothiopyranopyrimidine-2-ol (E) in potassium hydroxide; the intermediate E being hydrolyzed into tetrahydrothiopyrano pyrimidine-2,4-diol (F) in acetic acid; the intermediate F being base chlorinated with N, N-dimethylaniline into 2,4-dichloro tetrahydrothiopyranopyrimidine (G) in phosphorus oxychloride; the intermediate G being reacted by substituting phenol or aniline to give an intermediate H under alkaline condition; the intermediate H being reacted with the corresponding substituted aniline to obtain a final compound; or the intermediate H being reacted with 1-boc-4-aminopiperidine, deprotected with boc, then reacted with corresponding benzyl chloride or benzyl bromide to obtain the final compound; above reactions shown as the ##STR00053## ##STR00054## following: wherein reagents and conditions are (i) sodium methoxide, potassium iodide, methanol, reflux; (ii) sodium methoxide, toluene, 105 C.; (iii) methylisothiourea sulfate, potassium hydroxide, (V) phosphorus oxychloride, N, N-dimethylaniline at 90 C.; (vi) potassium carbonate, N, N-dimethylformamide, substituted phenol or aniline; 1-Boc-4-aminopiperidine, N, N-diisopropylethylamine, N-methylpyrrolidone at 120-130 C.; b) trifluoroacetic acid in dichloromethane at room temperature; c) (Viii) 1-Substituted-4-aminopiperidine, N, N-diisopropylethylamine, N-methylpyrrolidone, 130 C.; (ix) 4-aminobenzonitrile, 2,2-bis-(diphenylphosphino)-1,1-binaphthyl, tris (dibenzylideneacetone) dipalladium, dioxane; wherein, the 2,4,6-trisubstituted phenol/aniline is selected from a group consisting of 2,4,6-trimethylphenol, 4-hydroxy-3,5-dimethylbenzonitrile, (E)-3-(4-hydroxy-3,5-dimethylphenyl)acrylonitrile, 2,4,6-trimethylaniline, 4-amino-3,5-dimethylbenzonitrile, and (E)-3-(4-amino-3,5-dimethylphenyl)acrylonitrile; wherein, the substituted benzyl chloride/bromide is selected from a group consisting of 1-chloro-2-(chloromethyl)benzene, 1-chloro-3-(chloromethyl)benzene, 1-chloro-4-(chloromethyl)benzene, 1-bromo-2-(bromomethyl)benzene, 1-bromo-3-(bromomethyl)benzene, 1-bromo-4-(bromomethyl)benzene, 1-(chloromethyl)-2-fluorobenzene, 1-(chloromethyl)-3-fluorobenzene, 1-(chloromethyl)-4-fluorobenzene, 1-(bromomethyl)-2,4-difluorobenzene, 1-(bromomethyl)-3,4-difluorobenzene, 2-(chloromethyl)benzonitrile, 3-(chloromethyl)benzonitrile, 4-(chloromethyl)benzonitrile, 1-(cholomethyl)-2-nitrobenzene, 1-(chloromethyl)-3-nitrobenzene, 1-(chloromethyl)-4-nitrobenzene, 1-(chloromethyl)-2-methoxybenzene, 1-(chloromethyl)-3-methoxybenzene, 1-(chloromethyl)-4-methoxybenzene, 1-(bromomethyl)-4-(methylsulfonyl)benzene, 4-(bromomethyl)benzenesulfonamide, 3-(bromomethyl)benzenesulfonamide, 2-(bromomethyl)benzamide, N-(4-(bromomethyl)phenyl)formamide, ethyl 4-(bromomethyl)benzoate, 4-(bromomethyl)benzamide, 3-(bromomethyl)benzamide and N-(4-(bromomethyl)phenyl)methanesulfonamide; wherein A is S(O).sub.n or CH.sub.2; B is S(O).sub.n or CH.sub.2, while either A or B is S(O).sub.n in one compound; n=0, 1 or 2; X is O or NH; R.sup.1, R.sup.2 and R.sup.3 are independently selected from a group consisting of H, halo, CN, CF.sub.3, NH.sub.2, OH, C.sub.1-6alkyl, C.sub.1-6alkyloxy, C.sub.2-6alkenyl, trifluoromethyl, amino, hydroxy, cyanovinyl, cyanoethyl and cyclopropyl.

8. A method for preventing HIV infection or treating HIV-infected patient comprising a step of administrating to a subject in need a therapeutically effective amount of the compound of claim 1 or its pharmaceutically acceptable salt.

9. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 1 or its pharmaceutically acceptable salt and one or more pharmaceutical acceptable carrier or excipient.

Description

EXAMPLES

(1) Selected examples are listed as follows, the invention includes these compounds disclosed herein but not confined to them.

Example 1: Preparation of Intermediate 7

(2) ##STR00016##

Step A: methyl 4-((2-methoxy-2-oxoethyl)thio)butanoate (3)

(3) To a solution of sodium methoxide (2.75 g, 51 mmol) in MeOH (30 mL) was added methyl-2-mercaptoacetate (1, 7.85 mL, 87.8 mmol) and the reaction mixture was stirred at room temperature for 0.5 h. Potassium iodide (50 mg) and methyl-4-chlorobutanoate (2, 6.2 mL, 101 mmol) were added to the reaction mixture, which was stirred at 65 C. for 20 h. After cooling, the reaction mixture was filtered and the resultant filtrate was concentrated under reduced pressure. CH.sub.2Cl.sub.2 (100 mL) was added to the residue and the mixture was extracted with H.sub.2O (320 mL). The combined organic layer was washed with brine, dried over Na.sub.2SO.sub.4, and concentrated in vacuo to give methyl-4-((2-methoxy-2-oxoethyl)thio)butanoate (3) as a pale yellow oil. This compound was used for the next reaction without further purification. .sup.1H NMR (600 MHz, CDCl.sub.3) : 3.75 (s, 3H), 3.66 (s, 3H), 3.21 (s, 2H), 2.77 (t, J=7.2 Hz, 2H), 2.66 (t, J=7.2 Hz, 2H), 1.93 (p, J=7.2 Hz, 2H). ESI-MS: m/z 207.3 (M+1).sup.+, 224.4 (M+18).sup.+. C.sub.8H.sub.14O.sub.4S (206.06).

Step B: methyl 3-oxotetrahydro-2H-thiopyran-2-carboxylate (4)

(4) To a solution 3 (5.00 g, 24.3 mmol) in anhydrous toluene (40 mL) was added sodium methoxide (1.43 g, 26.5 mmol) and heated at 105 C. for 3 h. After cooling to room temperature, 12N HCl (5 mL) was added. The mixture was extracted with ethyl acetate (330 mL). The combined organic layers were washed with brine, then dried over Na.sub.2SO.sub.4, the filtrate was concentrated under reduced pressure and purified by silica-gel column chromatography to afford methyl-3-oxotetrahydro-2H-thiopyran-2-carboxylate (4) as a yellow oil (1.93 g, 45.8%). .sup.1H NMR (CDCl.sub.3) : 12.18 (1H, s), 2.79-2.83 (2H, m), 2.42 (2H, t, J=6.6 Hz), 2.10-2.17 (2H, m). ESI-MS: m/z 175.3 (M+1).sup.+, 192.5 (M+18).sup.+. C.sub.7H.sub.10O.sub.3S (174.04).

Step C: 2-(methylthio)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-ol (5)

(5) To a solution of KOH (0.57 g, 10.76 mmol) in MeOH (30 mL) were added 4 (1.25 g, 7.17 mmol) and S-methyisothiourea sulfate (1.00 g, 3.58 mmol). After stirring at room temperature for 16 h, the reaction mixture was poured into ice water (40 mL) and AcOH (2.0 mL) was added. The resultant precipitate was collected by filtration and washed with H.sub.2O to give 2-(methylthio)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-4-ol (5) as a white solid. This compound was used for the next reaction without further purification. mp 234-236 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 12.70 (s, 1H), 2.94-2.84 (m, 2H), 2.61 (t, J=6.3 Hz, 2H), 2.45 (s, 3H), 2.08-1.95 (m, 2H). ESI-MS: m/z 215.4 (M+1).sup.+, 237.3 (M+23).sup.+. C.sub.8H.sub.10N.sub.2OS.sub.2 (214.02).

Step D: 7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine-2,4-diol (6)

(6) To a solution of 2-(methylthio)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-ol 5 (1.05 g) in H.sub.2O (10 mL) was added acetic acid (20 mL) and the reaction mixture was stirred at 110 C. for 3 days. After cooling, the resultant precipitate was collected by filtration to give 7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine-2,4-diol (6, 0.5 g, 37.7% two steps) as a colorless crystal. mp>300 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 11.17 (s, 1H), 10.82 (s, 1H), 2.84 (d, J=6.0 Hz, 2H), 2.43 (t, J=6.3 Hz, 2H), 2.04-1.91 (m, 2H). ESI-MS: m/z 185.0 (M+1).sup.+, 207.2 (M+23).sup.+. C.sub.7H.sub.8N.sub.2O.sub.2S (184.03).

Step E: 2,4-dichloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine (7)

(7) A solution of 6 (0.42 g, 2.3 mmol) and N,N-dimethylaniline (70 L) in phosphoryl chloride (2.0 mL) was stirred at 90 C. for 18 h. After cooling, the reaction mixture was poured into ice water (50 mL) and the mixture was extracted with CH.sub.2Cl.sub.2 (30 mL3). The combined organic layer was washed with brine, dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was chromatographed to give 2,4-dichloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine (7) as a pale gray solid (0.35 g, 69.4%). mp 105-108 C. .sup.1H NMR (CDCl.sub.3) : 3.09-3.13 (2H, m), 2.99 (2H, t, J=6.3 Hz), 2.21-2.28 (2H, m). ESI-MS: m/z 221.3 (M+1).sup.+. C.sub.7H.sub.6Cl.sub.2N.sub.2S (219.96).

Example 2: Preparation of Intermediate 8

(8) ##STR00017##

4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (8a)

(9) To a solution of 7 (0.5 g, 2.26 mmol) in DMF were added K.sub.2CO.sub.3 (0.38 g, 2.71 mmol) and 2,4,6-trimethylphenol (0.40 g, 2.71 mmol), and stirred at room temperature overnight, the mixture was filtered and evaporated under reduce pressure. CH.sub.2Cl.sub.2 was added to the residue and the mixture was extracted with H.sub.2O. The organic layer was then washed with brine, dried over Na.sub.2SO.sub.4, concentrated under reduced pressure and recrystallized from ethyl acetate and petroleum ether to give 4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (8a, 0.54 g, 72.5%) as a pale grey solid. mp 254-255 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 7.73 (s, 2H), 3.16 (d, J=5.6 Hz, 2H), 2.92 (t, J=6.2 Hz, 2H), 2.23-2.13 (m, 2H), 2.07 (s, 6H). ESI-MS: m/z 332.4 (M+1).sup.+. C.sub.16H.sub.14ClN.sub.3OS (331.05).

2-chloro-4-(mesityloxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine (8b)

(10) Synthesized in a similar procedure with intermediate 7 using 2,4,6-trimethylphenol as starting material.

(11) Yield 72.5%, mp 176-179 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 7.05 (s, 2H), 3.04-2.93 (m, 2H), 2.66 (t, J=6.1 Hz, 2H), 2.24 (s, 3H), 2.09 (dt, J=12.1, 6.1 Hz, 2H), 1.98 (s, 6H). ESI-MS: m/z 321.4 (M+1).sup.+. C.sub.16H.sub.17ClN.sub.2OS (320.08).

(E)-3-(4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylphenyl)acrylonitrile (8c)

(12) Synthesized in a similar procedure with intermediate 7 using (E)-3-(4-hydroxy-3,5-dimethylphenyl)acrylonitrile as starting material.

(13) Yield 86.1%, mp 240-242 C. .sup.1H NMR (400 MHz, CDCl.sub.3) : 7.34 (d, J=16.7 Hz, 1H), 7.19 (s, 2H), 5.82 (d, J=16.6 Hz, 1H), 3.17-3.05 (m, 2H), 2.97 (t, J=6.3 Hz, 2H), 2.30 (p, J=6.2 Hz, 2H), 2.13 (s, 6H). ESI-MS: m/z 358.3 (M+1).sup.+. C.sub.18H.sub.16ClN.sub.3OS (357.07).

Example 3: Preparation of IA-1

(14) ##STR00018##

4-((2-((4-cyanophenyl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IA-1-1)

(15) ##STR00019##

(16) To a solution of 8a (0.50 g, 1.5 mmol) and 4-aminobenzonitrile (0.36 g, 3 mmol) in 1,4-dioxane ((20 mL), BINAP (93 mg, 0.15 mmol) and Pd.sub.2(dba).sub.3 (140 mg, 0.15 mmol) were added and stirred for 10 min; Cs.sub.2CO.sub.3 (0.98 g, 3 mmol) was added and the mixture was heated to 100 C. under N.sub.2 atmosphere overnight. Residue was filtered and evaporated under reduced pressure. Ethyl acetate (100 mL) was added and extracted with H.sub.2O (320 mL), organic layer was washed with brine, dried with Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude material was purified by column chromatography over silica gel (200-300 mesh) with MeOH/CH.sub.2Cl.sub.2 as an eluent to give 4-((2-((4-cyanophenyl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl) oxy)-3,5-dimethylbenzonitrile (IA-1-1) as a white solid (0.48 g, 77.0%). mp 253-254 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 9.98 (s, 1H), 7.78 (s, 2H), 7.45 (s, 4H), 3.19-3.05 (m, 2H), 2.88 (t, J=6.2 Hz, 2H), 2.26-2.16 (m, 2H), 2.11 (s, 6H). .sup.13C NMR (100 MHz, DMSO) : 163.63, 163.07, 154.68, 154.12, 145.20, 133.10, 133.02, 132.98, 119.92, 119.00, 118.05, 109.09, 104.50, 102.39, 31.74, 26.34, 22.96, 16.12. ESI-MS: m/z 414.5 (M+1).sup.+, 431.5 (M+18).sup.+. C.sub.23H.sub.19N.sub.5OS (413.13).

(E)-4-((4-(4-(2-cyanovinyl)-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-yl)amino)benzonitrile (IA-1-2)

(17) ##STR00020##

(18) Synthesized in a similar procedure with example 3 using (E)-3-(4-((2-chloro-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylphenyl)acrylonitrile as starting material.

(19) Yield 64.3%, mp 278-280 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 9.95 (s, 1H), 7.68 (d, J=16.7 Hz, 1H), 7.54 (s, 2H), 7.48 (d, J=8.6 Hz, 2H), 7.39 (d, J=8.8 Hz, 2H), 6.48 (d, J=16.7 Hz, 1H), 3.19-3.06 (m, 2H), 2.87 (t, J=6.2 Hz, 2H), 2.20 (p, J=6.0 Hz, 2H), 2.08 (s, 6H). .sup.13C NMR (100 MHz, DMSO) : 163.47, 163.30, 154.76, 152.47, 150.45, 145.29, 132.97, 131.93, 131.63, 128.64, 119.95, 119.33, 118.06, 104.49, 102.26, 96.93, 31.71, 26.33, 22.98, 16.40. ESI-MS: m/z 440.6 (M+1).sup.+, 457.6 (M+18).sup.+. C.sub.25H.sub.21N.sub.5OS (439.15).

4-((2-((4-cyanophenyl)amino)-5-oxido-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IA-1-3)

(20) ##STR00021##

(21) Compound IA-1-1 (200 mg, 0.48 mmol) was dissolved in CH.sub.2Cl.sub.2 (5 mL) and cooled to 78 C., a mixture of mCPBA (107 mg, 0.57 mmol) in CH.sub.2Cl.sub.2 (3 mL) was added and stirred for 1 h. Residue was heated to room temperature and NaHSO.sub.3 (30 mL) was added, extracted with ethyl acetate (20 mL3). Combined organic layer and washed with brine, dried with Na.sub.2SO.sub.4 and concentrated under reduced pressure. Recrystallized from MeOH/CHCl.sub.3 to give compound IA-1-3 (150 mg, 72.2%) as a white solid. mp 247-249 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 10.52 (s, 1H), 7.80 (d, J=4.6 Hz, 2H), 7.50 (s, 4H), 3.32-3.19 (m, 1H), 3.09-2.79 (m, 3H), 2.61-2.51 (m, 2H), 2.15 (d, J=14.7 Hz, 6H). .sup.13C NMR (100 MHz, DMSO) : 169.09, 167.61, 158.94, 153.63, 144.12, 133.16, 133.08, 119.60, 119.31, 118.95, 111.11, 109.44, 104.19, 60.21, 45.19, 32.08, 16.22, 12.89. ESI-MS: m/z 447.5 (M+18).sup.+, 452.3 (M+23).sup.+. C.sub.23H.sub.19N.sub.5O.sub.2S (429.13).

4-((2-((4-cyanophenyl)amino)-5,5-dioxido-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IA-1-4)

(22) ##STR00022##

(23) Compound IA-1-1 (100 mg, 0.24 mmol) was dissolved in CH.sub.2Cl.sub.2 (10 mL) and a mixture of mCPBA (125 mg, 0.73 mmol) was added at room temperature and stirred for 2 h. CH.sub.2Cl.sub.2 (50 mL) was added and washed with NaHSO.sub.3 (30 mL) and H.sub.2O (20 mL2). Organic layer was washed with brine, dried with Na.sub.2SO.sub.4 and concentrated under reduced pressure. Recrystallized from MeOH/CHCl.sub.3 to give compound IA-1-4 (67 mg, 62.2%) as a colorless crystal. mp 263-265 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 10.62 (s, 1H), 7.81 (s, 2H), 7.64-7.18 (m, 4H), 3.69-3.55 (m, 2H), 3.02 (t, J=6.0 Hz, 2H), 2.34 (d, J=5.1 Hz, 2H), 2.16 (s, 6H). .sup.13C NMR (100 MHz, DMSO) : 169.89, 164.70, 158.49, 153.31, 143.82, 133.19, 133.08, 132.92, 119.52, 119.44, 118.92, 112.61, 109.52, 104.48, 52.49, 32.08, 18.98, 16.08. ESI-MS: m/z 463.5 (M+18).sup.+, 468.4 (M+23).sup.+. C.sub.23H.sub.19N.sub.5O.sub.3S (445.12).

Example 4. Preparation of Intermediate 9

3,5-dimethyl-4-((2-(piperidin-4-ylamino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (9a)

(24) ##STR00023##

(25) To a solution of 8a (2.00 g, 6.03 mmol) and N-Boc-4-piperidineamine (1.5 equiv) in NMP (10 mL), DIPEA (1.2 mL) was added and stirred at 100-120 C. for 4-6 h. After cooling, ethyl acetate was added to the reaction mixture. The ethyl acetate layer was washed with H.sub.2O, dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The residue was added TFA/CH.sub.2Cl.sub.2 (1:1) and stirred at room temperature for 4 hours. Saturated aqueous sodium bicarbonate was added to the reaction mixture to neutralize the residues. Ethyl acetate was added and organic layer was separated, washed with H.sub.2O, dried over sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography over silica gel (200-300) with CH.sub.2Cl.sub.2/MeOH as an eluent to yield 3,5-dimethyl-4-((2-(piperidin-4-ylamino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (9a) as a white powder (1.5 g, 62.9%). mp>250 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 9.00 (s, 2H), 7.67 (s, 2H), 7.02 (s, 1H), 3.14 (d, J=12.5 Hz, 2H), 3.07-2.99 (m, 2H), 2.71 (t, J=6.3 Hz, 4H), 2.18-2.09 (m, 2H), 2.07 (s, 6H), 1.81 (s, 2H), 1.54 (s, 2H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) : 163.27, 157.76, 154.08, 132.96, 132.77, 119.08, 108.60, 46.02, 41.99, 31.86, 28.05, 26.31, 23.39, 16.18. ESI-MS: m/z 396.4 (M+1).sup.+. C.sub.21H.sub.25N.sub.5OS (395.18).

4-(mesityloxy)-N-(piperidin-4-yl)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-amine (9b)

(26) Synthesized in a similar procedure with intermediate 9a using 8b as starting material. Yield 28.7%, mp>290 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 8.91 (s, 2H), 6.90 (s, 3H), 3.15 (d, J=12.6 Hz, 2H), 3.07-2.96 (m, 2H), 2.69 (t, J=6.3 Hz, 4H), 2.24 (s, 3H), 2.12 (p, J=5.9 Hz, 2H), 1.98 (s, 6H), 1.82 (s, 2H), 1.54 (s, 2H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) : 164.00, 157.90, 147.95, 134.59, 130.15, 129.29, 46.06, 42.15, 31.84, 28.14, 26.30, 23.45, 20.81, 16.44. ESI-MS: m/z 385.5 (M+1).sup.+. C.sub.21H.sub.28N.sub.4OS (384.20).

(E)-3-(3,5-dimethyl-4-((2-(piperidin-4-ylamino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)phenyl)acrylonitrile (9c)

(27) Synthesized in a similar procedure with intermediate 9a using 8c as starting material. Yield 54.2%, mp>280 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 8.85 (s, 2H), 7.59 (d, J=16.7 Hz, 1H), 7.43 (s, 2H), 7.30 (s, 2H), 6.66 (s, 1H), 6.40 (d, J=16.7 Hz, 1H), 3.43 (s, 2H), 3.08-2.95 (m, 2H), 2.68 (t, J=6.2 Hz, 4H), 2.15-2.09 (m, 2H), 2.06 (s, 1H), 2.03 (s, 6H), 1.61 (s, 4H), 1.32 (s, 2H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) : 163.65, 158.05, 152.59, 150.58, 131.62, 131.45, 126.05, 119.42, 96.51, 62.02, 52.67, 49.07, 31.89, 26.34, 23.50, 16.46. ESI-MS: m/z 422.5 (M+1).sup.+. C.sub.23H.sub.27N.sub.5OS (421.19).

Example 5. Preparation of Compounds IIA-1

(28) ##STR00024##

4-((4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-yl)amino)piperidin-1-yl)methyl)benzenesulfonamide (IIA-1-1)

(29) ##STR00025##

(30) To a solution of 9a (0.20 g, 0.51 mmol) in DMF (10 mL), K.sub.2CO.sub.3 (0.19 g, 0.76 mmol) and 4-(bromomethyl)benzenesulfonamide (0.11 g, 0.76 mmol) were added and stirred overnight. The reaction mixture was concentrated under reduced pressure. Ethyl acetate was added to the reaction mixture. The ethyl acetate layer was washed with H.sub.2O, dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude material was purified by column chromatography over silica gel (200-300 mesh) with CH.sub.2Cl.sub.2/MeOH as an eluent and washed with MeOH to afford the corresponding compounds as white powder (0.24 g, 84.1%). mp 193-195 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.77 (d, J=8.3 Hz, 2H), 7.66 (s, 2H), 7.45 (d, J=8.0 Hz, 2H), 7.30 (s, 2H), 6.88 (s, 1H), 3.45 (s, 2H), 3.13-2.95 (m, 2H), 2.91-2.57 (m, 4H), 2.16-2.08 (m, 2H), 2.06 (s, 6H), 1.99-0.81 (m, 6H). .sup.13C NMR (100 MHz, DMSO) 163.25, 157.97, 154.22, 143.35, 143.13, 133.02, 132.66, 129.41, 126.04, 119.10, 108.56, 61.97, 52.61, 49.07, 31.86, 31.65, 26.34, 23.48, 16.15. ESI-MS: m/z 565.5 (M+1).sup.+. C.sub.28H.sub.32N.sub.6O.sub.3S.sub.2 (564.20).

(31) Synthesized of IIA-1-2-IIA-1-13 in a similar procedure to IIA-1-1 using the appropriate starting material.

4-((4-((4-(mesityloxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-yl)amino) piperidin-1-yl)methyl)benzenesulfonamide (IIA-1-2)

(32) ##STR00026##

(33) Starting with 9b (0.25 g, 0.65 mmol) and 4-(bromomethyl)benzenesulfonamide (0.20 mg, 0.78 mmol) to afford IIA-1-2 (157 mg, 43.6%) as a white powder. mp 218-220 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.77 (d, J=8.3 Hz, 2H), 7.44 (d, J=8.0 Hz, 2H), 7.30 (s, 2H), 6.88 (s, 2H), 6.60 (s, 1H), 3.45 (s, 2H), 3.08-2.91 (m, 2H), 2.79-2.57 (m, 4H), 2.23 (s, 3H), 2.10 (dt, J=11.8, 5.4 Hz, 2H), 1.97 (s, 6H), 1.72-1.47 (m, 2H), 1.46-1.10 (m, 2H). .sup.13C NMR (100 MHz, DMSO) 163.97, 162.36, 158.15, 148.02, 143.36, 143.13, 134.49, 130.15, 129.42, 129.22, 126.03, 61.99, 52.65, 49.07, 31.86, 31.74, 26.33, 23.54, 20.80, 16.43. ESI-MS: m/z 554.5 (M+1).sup.+. C.sub.28H.sub.35N.sub.5O.sub.3S.sub.2 (553.22).

(E)-4-((4-((4-(4-(2-cyanovinyl)-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-yl)amino)piperidin-1-yl)methyl)benzenesulfonamide (IIA-1-3)

(34) ##STR00027##

(35) Starting with 9c (0.20 g, 0.47 mmol) and 4-(bromomethyl)benzenesulfonamide (0.14 g, 0.57 mmol) to afford IIA-1-3 (0.18 g, 64.2%) as a white powder. mp 237-239 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.77 (d, J=8.3 Hz, 2H), 7.59 (d, J=16.7 Hz, 1H), 7.43 (s, 4H), 7.30 (s, 2H), 6.66 (s, 1H), 6.40 (d, J=16.7 Hz, 1H), 3.43 (s, 2H), 3.08-2.95 (m, 2H), 2.68 (t, J=6.2 Hz, 4H), 2.15-2.09 (m, 2H), 2.06 (s, 1H), 2.03 (s, 6H), 1.61 (s, 4H), 1.32 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.65, 158.05, 152.59, 150.58, 143.41, 143.14, 131.62, 131.45, 129.43, 128.33, 126.05, 119.42, 96.51, 62.02, 52.67, 49.07, 31.89, 31.68, 26.34, 23.50, 16.46. ESI-MS: m/z 591.2257 (M+1).sup.+. C.sub.30H.sub.34N.sub.6O.sub.3S.sub.2 (590.2134).

3-((4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-2-yl)amino)piperidin-1-yl)methyl)benzenesulfonamide (IIA-1-4)

(36) ##STR00028##

(37) Starting with 9a (0.20 g, 0.51 mmol) and 3-(bromomethyl)benzenesulfonamide (0.15 g, 0.61 mmol) to afford IIA-1-4 (0.21 g, 73.5%) as a white solid. mp 170-172 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.74 (s, 1H), 7.71 (d, J=7.4 Hz, 1H), 7.67 (s, 2H), 7.51 (t, J=7.5 Hz, 1H), 7.47 (d, J=7.5 Hz, 1H), 7.36 (s, 2H), 6.80 (s, 1H), 3.46 (s, 2H), 3.02 (dd, J=6.7, 4.1 Hz, 2H), 2.69 (t, J=6.1 Hz, 4H), 2.19-2.09 (m, 2H), 2.08 (s, 1H), 2.06 (s, 6H), 1.66 (s, 4H), 1.32 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.25, 157.84, 154.22, 144.59, 144.09, 140.31, 133.01, 132.74, 132.47, 129.26, 125.92, 124.68, 119.13, 108.55, 62.05, 52.62, 49.07, 31.82, 31.65, 26.30, 23.45, 16.18. ESI-MS: m/z 565.2045 (M+1).sup.+. C.sub.28H.sub.32N.sub.6O.sub.3S.sub.2 (564.1977).

4-((2-((1-(4-cyanobenzyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IIA-1-5)

(38) ##STR00029##

(39) Starting with 9a (0.20 g, 0.51 mmol) and 4-(chloromethyl)benzonitrile (92 mg, 0.61 mmol) to afford IIA-1-5 (0.21 g, 81.3%) as a white solid. mp 184-186 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.77 (d, J=8.2 Hz, 2H), 7.66 (s, 2H), 7.47 (d, J=8.1 Hz, 2H), 6.89 (br, 1H), 3.48 (s, 2H), 3.12-2.95 (m, 2H), 2.69 (t, J=6.3 Hz, 5H), 2.19-2.02 (m, 8H), 1.63 (s, 4H), 1.32 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.26, 158.00, 154.22, 145.26, 133.01, 132.66, 132.58, 129.87, 119.38, 119.10, 110.08, 108.56, 61.93, 52.59, 48.66, 31.84, 31.66, 26.33, 23.47, 16.16. ESI-MS: m/z 511.6 (M+1).sup.+. C.sub.29H.sub.30N.sub.6OS (510.22).

4-((2-((1-(3-cyanobenzyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IIA-1-6)

(40) ##STR00030##

(41) Starting with 9a (0.20 g, 0.51 mmol) and 4-(bromomethyl)benzonitrile (119 mg, 0.61 mmol) to afford IIA-1-6 (0.21 g, 81.3%) as a white solid. mp 164-166 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.72 (d, J=7.6 Hz, 1H), 7.68 (s, 1H), 7.65 (s, 2H), 7.62 (d, J=7.8 Hz, 1H), 7.53 (t, J=7.7 Hz, 1H), 6.87 (s, 1H), 3.45 (s, 2H), 3.09-2.94 (m, 2H), 2.69 (t, J=6.3 Hz, 4H), 2.19-2.09 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.63 (s, 4H), 1.31 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.26, 157.96, 154.22, 140.90, 134.02, 133.02, 132.68, 132.44, 131.22, 129.89, 119.34, 119.10, 111.65, 108.56, 61.46, 55.38, 52.49, 31.86, 31.64, 31.15, 26.33, 23.47, 21.52, 16.16. ESI-MS: m/z 511.2335 (M+1).sup.+. C.sub.29H.sub.30N.sub.6OS (510.2202).

4-((2-((1-(2-cyanobenzyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IIA-1-7)

(42) ##STR00031##

(43) Starting with 9a (0.20 g, 0.51 mmol) and 4-(chloromethyl)benzonitrile (92 mg, 0.61 mmol) to afford IIA-1-7 (0.20 g, 77.5%) as a white crystal. mp 206-208 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.79 (d, J=7.6 Hz, 1H), 7.70-7.61 (m, 3H), 7.52 (d, J=7.7 Hz, 1H), 7.46 (t, J=7.6 Hz, 1H), 6.86 (s, 1H), 3.56 (s, 2H), 3.10-2.94 (m, 2H), 2.69 (t, J=6.2 Hz, 4H), 2.17-2.09 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.59 (s, 4H), 1.30 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.26, 158.00, 154.22, 142.82, 133.42, 133.41, 133.02, 132.69, 130.48, 128.36, 119.08, 118.11, 112.53, 108.57, 60.36, 60.22, 52.54, 31.88, 31.58, 31.19, 26.33, 23.47, 16.17, 14.56. ESI-MS: m/z 511.3 (M+1).sup.+. C.sub.29H.sub.30N.sub.6OS (510.22).

4-((4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-yl)amino)piperidin-1-yl)methyl)benzamide (IIA-1-8)

(44) ##STR00032##

(45) Starting with 9a (0.20 g, 0.51 mmol) and 4-(chloromethyl)benzamide (103 mg, 0.61 mmol) to afford IIA-1-8 (0.20 g, 74.8%) as a white crystal. mp 261-263 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.91 (s, 1H), 7.81 (d, J=8.1 Hz, 2H), 7.66 (s, 2H), 7.33 (d, J=7.9 Hz, 2H), 7.30 (s, 1H), 6.86 (br, 1H), 3.43 (s, 2H), 3.08-2.94 (m, 2H), 2.69 (t, J=6.2 Hz, 4H), 2.17-2.10 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.60 (s, 4H), 1.31 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 168.25, 163.26, 158.04, 154.23, 142.60, 142.52, 133.45, 133.02, 132.69, 128.88, 127.88, 119.11, 108.56, 79.65, 62.19, 52.63, 31.84, 31.65, 31.16, 26.33, 23.47, 16.16. ESI-MS: m/z 529.5 (M+1).sup.+, 551.6 (M+23).sup.+. C.sub.29H.sub.32N.sub.6O.sub.2S (528.23).

3-((4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-2-yl)amino)piperidin-1-yl)methyl)benzamide (IIA-1-9)

(46) ##STR00033##

(47) Starting with 9a (0.20 g, 0.51 mmol) and 3-(chloromethyl)benzamide (103 mg, 0.61 mmol) to afford IIA-1-9 (0.19 g, 71.1%) as a white solid. mp 245-247 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.95 (s, 1H), 7.76 (d, J=11.6 Hz, 2H), 7.66 (s, 2H), 7.39 (d, J=7.3 Hz, 2H), 7.33 (s, 1H), 6.86 (s, 1H), 3.43 (s, 2H), 3.02 (s, 2H), 2.77-2.58 (m, 4H), 2.19-2.09 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.63 (s, 4H), 1.32 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 168.43, 163.26, 157.96, 154.22, 139.22, 134.68, 133.02, 132.68, 132.03, 128.45, 126.44, 119.10, 108.56, 62.44, 52.60, 48.81, 31.87, 31.65, 31.15, 26.33, 23.47, 16.16. ESI-MS: m/z 529.4 (M+1).sup.+, 551.6 (M+23).sup.+. C.sub.29H.sub.32N.sub.6O.sub.2S (528.23).

ethyl 4-((4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-2-yl)amino)piperidin-1-yl)methyl)benzoate (IIA-1-10)

(48) ##STR00034##

(49) Starting with 9a (0.2 g, 0.51 mmol) and ethyl 4-(chloromethyl)benzoate (0.15 g, 0.61 mmol) to afford IIA-1-10 (0.16 g, 56.7%) as a white solid. mp 181-183 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.91 (d, J=8.1 Hz, 2H), 7.65 (s, 2H), 7.41 (d, J=8.1 Hz, 2H), 6.87 (br, 1H), 4.30 (q, J=7.1 Hz, 2H), 3.46 (s, 2H), 3.09-2.97 (m, 2H), 2.69 (t, J=6.2 Hz, 4H), 2.16-2.09 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.60 (s, 4H), 1.31 (t, J=7.1 Hz, 5H). .sup.13C NMR (100 MHz, DMSO) 166.11, 163.26, 157.99, 154.24, 144.87, 141.04, 133.02, 132.74, 129.52, 129.29, 128.98, 119.10, 108.56, 62.16, 61.04, 56.50, 52.62, 31.88, 31.65, 31.16, 26.33, 23.47, 16.15, 14.66. ESI-MS: m/z 558.6 (M+1).sup.+. C.sub.31H.sub.35N.sub.5O.sub.3S (557.25).

3,5-dimethyl-4-((2-((1-(4-(methylsulfonyl)benzyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (IIA-1-11)

(50) ##STR00035##

(51) Starting with 9a (0.20 g, 0.51 mmol) and 1-(chloromethyl)-4-(methylsulfonyl)benzene (0.15 g, 0.61 mmol) to afford IIA-1-11 (0.19 g, 66.7%) as a white solid. mp 265-267 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.87 (d, J=8.3 Hz, 2H), 7.66 (s, 2H), 7.54 (d, J=8.2 Hz, 2H), 6.89 (br, 1H), 3.50 (s, 2H), 3.20 (s, 3H), 3.10-2.95 (m, 2H), 2.69 (t, J=6.2 Hz, 4H), 2.11 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.64 (s, 4H), 1.32 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.26, 158.00, 157.98, 154.22, 145.46, 139.81, 133.02, 132.69, 129.77, 127.39, 119.12, 108.56, 61.89, 52.65, 48.68, 44.07, 31.86, 31.66, 26.33, 23.47, 16.16. ESI-MS: m/z 564.5 (M+1).sup.+, 586.5 (M+23).sup.+. C.sub.29H.sub.33N.sub.5O.sub.3S.sub.2 (563.20).

3,5-dimethyl-4-((2-((1-(4-nitrobenzyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)benzonitrile (IIA-1-12)

(52) ##STR00036##

(53) Starting with 9a (0.20 g, 0.51 mmol) and 1-(chloromethyl)-4-nitrobenzene (0.13 g, 0.61 mmol) to afford IIA-1-12 (0.14 g, 52.2%) as a white solid. mp 190-192 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.18 (d, J=8.7 Hz, 2H), 7.66 (s, 2H), 7.55 (d, J=8.6 Hz, 2H), 6.88 (s, 1H), 3.53 (s, 2H), 3.11-2.92 (m, 2H), 2.69 (t, J=6.2 Hz, 4H), 2.19-2.10 (m, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.59 (s, 4H), 1.33 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.26, 157.99, 154.23, 147.51, 146.97, 133.02, 132.67, 130.03, 123.80, 119.10, 108.56, 61.63, 52.61, 31.84, 31.66, 31.15, 26.33, 23.47, 19.03, 16.15. ESI-MS: m/z 531.5 (M+1).sup.+. C.sub.28H.sub.30N.sub.6O.sub.3S (530.21).

Example 6. Preparation of 4-((2-((1-(4-aminobenzyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IIA-1-13)

(54) ##STR00037##

(55) To a solution of Fe powder (0.56 g, 10 mmol) and NH.sub.4Cl (55 mg, 1 mmol) in H.sub.2O (5 mL) was added AcOH (0.12 mL), the mixture was heated to 50 C. for 15 min. A solution of IIA-1-12 (0.53 g, 1.0 mmol) in DMF (5 mL) was added and stirred for another 15 min. After cooling to the room temperature, pH was adjusted to >9 with NaHCO.sub.3, and residue was filtered over diatomite. H.sub.2O (50 mL) was added to the filtrate and extracted with ethyl acetate (20 mL3) and the combined organic layer was washed with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The residue was purified by silica gel column chromatography using MeOH/CH.sub.2Cl.sub.2 as an eluent to afford IIA-1-13 (0.31 g, 63.6%) as a white solid. mp 169-171 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.66 (s, 2H), 6.87 (d, J=8.1 Hz, 2H), 6.49 (d, J=8.2 Hz, 2H), 4.94 (s, 2H), 3.17 (s, 2H), 3.08-2.95 (m, 2H), 2.68 (t, J=6.1 Hz, 4H), 2.12 (d, J=5.3 Hz, 2H), 2.09 (s, 1H), 2.05 (s, 6H), 1.48 (s, 4H), 1.23 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.24, 158.03, 154.22, 147.95, 133.01, 132.65, 130.16, 125.66, 123.82, 119.11, 114.01, 108.55, 62.55, 52.39, 31.65, 26.31, 23.46, 21.24, 19.03, 16.16, 14.56. ESI-MS: m/z 501.5 (M+1).sup.+. C.sub.28H.sub.32N.sub.6OS (500.24).

Example 7. Preparation of N-(4-((4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidin-2-yl)amino)piperidin-1-yl)methyl)phenyl) methanesulfonamide (IIA-1-14)

(56) ##STR00038##

(57) To a solution of IIA-1-13 (100 mg, 0.2 mmol) and Et.sub.3N (61 mg, 0.6 mmol) in CH.sub.2Cl.sub.2 in an ice bath was added a solution of methanesulfonyl chloride (69 mg, 0.6 mmol) and stirred for 4 h. H.sub.2O (15 mL) was added to the solution and pH was adjusted to >7 with NaHCO.sub.3, then extracted with ethyl acetate (15 mL3) and the combined organic layer was washed with brine, dried over Na.sub.2SO.sub.4 and evaporated to dryness. The crude product was recrystallized from MeOH/CHCl.sub.3 to afford IIA-1-14 (20 mg, 17.3%) as white solid. mp 153-157 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.68 (s, 1H), 7.66 (s, 2H), 7.21 (d, J=8.3 Hz, 2H), 7.14 (d, J=8.4 Hz, 2H), 6.70 (br, 1H), 3.32 (s, 2H), 3.07-2.99 (m, 2H), 2.96 (s, 3H), 2.69 (t, J=6.3 Hz, 4H), 2.11 (t, J=5.7 Hz, 2H), 2.09 (s, 1H), 2.06 (s, 6H), 1.55 (s, 4H), 1.23 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.25, 137.49, 134.60, 133.01, 132.90, 132.77, 132.70, 130.12, 120.24, 119.13, 108.54, 100.98, 62.08, 52.56, 39.62, 36.25, 31.81, 31.63, 31.17, 26.30, 23.45, 16.16. ESI-MS: m/z 579.2250 (M+1).sup.+. C.sub.29H.sub.34N.sub.6O.sub.3S.sub.2 (578.2134).

Example 8. Preparation of IIC-1

4-((2-((1-(3-cyanophenyl)piperidin-4-yl)amino)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-4-yl)oxy)-3,5-dimethylbenzonitrile (IIC-1-1)

(58) ##STR00039##

(59) To a solution of 8a (0.20 g, 0.6 mmol) and 3-(4-aminopiperidin-1-yl)benzonitrile (145 mg, 0.72 mmol) in NMP (5 mL), DIPEA (120 L) was added, the mixture was then heated to 100 C. overnight. The mixture was cooled to room temperature; H.sub.2O was added and stirred for 30 min. The product was filtered to recrystallized from MeOH/CHCl.sub.3 to give the white solid (129 mg, 43.1%). mp 186-187 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.65 (s, 2H), 7.34 (dd, J=8.5, 7.4, 1H), 7.26 (s, 1H), 7.21 (d, J=8.5 Hz, 1H), 7.09 (d, J=7.4 Hz, 1H), 6.82 (br, 1H), 3.67 (s, 3H), 3.11-2.94 (m, 2H), 2.70 (t, J=6.0 Hz, 3H), 2.12 (t, J=5.7 Hz, 2H), 2.08 (s, 1H), 2.07 (s, 6H), 1.69 (s, 2H), 1.35 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 163.26, 157.91, 154.17, 151.25, 133.01, 132.73, 130.64, 121.53, 120.26, 119.84, 119.12, 118.11, 112.37, 108.58, 56.52, 47.19, 31.85, 31.16, 30.82, 26.31, 23.42, 18.98, 16.18. ESI-MS: m/z 497.2133 (M+1).sup.+. C.sub.28H.sub.28N.sub.6OS (496.2045).

3-(4-((4-(4-cyano-2,6-dimethylphenoxy)-7,8-dihydro-6H-thiopyrano[3,2-d] pyrimidin-2-yl)amino)piperidin-1-yl)benzamide (IIC-1-2)

(60) ##STR00040##

(61) Synthesized of IIC-1-2 in a similar procedure with IIC-1-1 using the 3-(4-aminopiperidin-1-yl)benzamide as starting material. Yield 35.1%, mp 232-234 C. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.91 (s, 1H), 7.68 (s, 2H), 7.38 (s, 1H), 7.29 (s, 1H), 7.25 (s, 1H), 7.23 (s, 1H), 7.03 (d, J=4.1 Hz, 2H), 6.73 (s, 1H), 3.65 (s, 2H), 3.03 (d, J=5.4 Hz, 2H), 2.72 (t, J=6.3 Hz, 3H), 2.13 (t, J=5.7 Hz, 2H), 2.09 (s, 1H), 2.09 (s, 6H), 1.72 (s, 2H), 1.42 (s, 2H). .sup.13C NMR (100 MHz, DMSO) 168.71, 163.27, 157.95, 154.18, 151.18, 135.40, 133.01, 132.75, 129.25, 119.11, 118.66, 117.98, 115.09, 108.58, 56.50, 48.06, 31.88, 31.16, 26.32, 23.46, 19.04, 16.21. ESI-MS: m/z 515.2226 (M+1).sup.+. C.sub.28H.sub.30N.sub.6O.sub.2S (514.2151).

Example 9. In Vitro Anti-HIV Activity of Compounds

(62) Selected compounds were screened for inhibitory activity against HIV-1 using MTT method as describe previously by Christophe. Pannecouque et al. Nat. Protoc. 3 (2008) 427-434, and Rudi Pauwels et al. J. Virol. Methods 20 (1988) 309-321. Briefly, MT-4 cells infected with HIV-1 can only survive for 5 to 7 days without any treatment, but when HIV-1 inhibitors were added, they can protect MT-4 cell from cytopathic. Serial solution of compounds was added to MT-4 cells after infected with HIV-1, MTT method was used to detect the survival rate after culture for 5 to 7 days. EC.sub.50 value was defined as compound concentration required to achieve 50% protection of MT-4 cells against HIV-1-induced cytopathic effect. In vitro anti-HIV-1 assay was supported by Rega Institute for Medical Research.

(63) 1. Materials:

(64) (1) MT-4 cells infected with HIV-1 viral strains (IIIB, L100I, K103N, Y181C, Y188L, E138K, F227L/V106A, Y181C/K103N) were provided by Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium. (2) MTT and formazan: sigma Chemical Co. (3) Preparation of compounds: Stock solutions (10final concentration) of test compounds is diluted with double distilled water for 5 folds and 5 concentrations of one compound are prepared. (4) Reference drugs: Nevirapine (NVP), Efavirenz (EFV), Etravirine (ETV), Rilpivirine (RPV) and Azidothymidine (AZT). (5) Method (MTT method): Serial five-fold dilutions of test compounds were added to cultured MT-4 cells infected with HIV-1, after 5 to 7 days, MTT was added and cultured for a few hours. Medium was removed and lysate was added followed by formazan, OD value was determined in 690 nm and 540 nm by microplate reader, and EC.sub.50 value was calculated.
2. Method

(65) The MTT method was described briefly as follows: 96-well plastic microtiter trays were filled with 100 L of complete medium. Subsequently, serial of tested compounds was added (25 L) to two series of triplicate wells so as to allow simultaneous evaluation of their effects on HIV- and mock infected cells. 50 L of 110.sup.4 cells/mL MT-4 cells were added. After cultured for 5 days at 37 C. in humidified atmosphere in the presence of 5% CO.sub.2, MTT was added and cultured for another 2 h, then medium was removed and 100 L isopropanol solution was added to lyse the cells. Formazan crystals were added and vibrated platform shaker for 10 min to solubilize the formazan crystals. Absorbances at 690 nm and 540 nm were read by using spectrophotometrically. EC.sub.50 was defined as the concentration achieving 50% protection from the cytopathic effect of the virus in infected cells.

(66) 2. Biological Data for Selected Compounds

(67) Selected compounds prepared as described above were assayed in MT-4 cells infected with wide-type III.sub.B, L100I, K103N, Y181C, Y188L, E138K or double mutant RES056 (Y181C/K103N) and F227L/V106A HIV-1 strains. Nevirapine, Efavirenz, Etravirine, Rilpivirine and Azidothymidine were selected as reference drugs. The results were listed below as EC.sub.50 value (nM)

(68) TABLE-US-00001 EC.sub.50 (nM) Compds wt L100I K103N Y181C Y188L E138K F227L/V106A Y181C/K103N IA-1-1 A B A B B B B B IA-1-2 A A A B B B B B IA-1-3 A C B B B B C C IA-1-4 A B A B B B B B IIA-1-1 A B A B A A B B IIA-1-2 A B A B B B B C IIA-1-3 A A A A B B A B IIA-1-4 A C B B B B B C IIA-1-5 A B B B B B C C IIA-1-6 A C B B B B C C IIA-1-7 A C B B C B C C IIA-1-8 A B A B B B C B IIA-1-9 A B B B B B C C IIA-1-10 B C B C C C C C IIA-1-11 A B A B B B B B IIA-1-12 A C B C B B C C IIA-1-13 A B A B B B C C IIA-1-14 A B A A B A C C IIC-1-1 B C B C C C C C IIC-1-2 A B A A A B C B NVP C C C C C C C C EFV A B B A B A B C ETV A A A B B B B B RPV A A A A B A B B AZT A A A A A A A B Table legend: EC.sub.50: Concentration of compound required to achieve 50% protection of MT-4 cells against HIV-1-induced cytopathicity. A is <10 nM; B is 10-100 nM; C is >100 nM.
3. Conclusion

(69) As indicated from the anti-HIV results, the invented tetrahydrothiopyranopyrimidine derivatives possessed a novel scaffold and excellent antiviral activities. Inhibitory activity of these compounds was less than 1 M against both wide-type and mutant HIV-1, among which IA-1-4 demonstrated the best activity. Activity against wide-type HIV-1 of IA-1-4 was 30 folds better than NVP and comparable with second generation NNRTIs RPV. When it came to mutant HIV-1 strains, IIA-1-3 showed the best performance and indicated comparable activity against K103N and Y188L with ETV, and 2-fold more potent against L100I, Y181C and double mutant strains (F227L/V106A and Y181C/K103N) than ETV. Therefore, tetrahydrothiopyranopyrimidine derivatives were potent NNRTIs and can be further developed as anti-HIV agents.