Sulfonamide benzamide derivative and preparation method and application thereof

Abstract

A sulfonamide benzamide derivative has the structure shown in Formula II that can be prepared as an anti-HBV (hepatitis B virus) pharmaceutical composition; it is prepared by sulfonation reaction, acylation reaction and sulfonylation reaction; the sulfonamide benzamide derivative has been shown effective activity for anti-HBV by inhibiting HBV DNA replication in vitro.

Claims

1. A sulfonamide benzamide derivative having the structure shown in general formula II as follows; ##STR00029## wherein R.sub.1-R.sub.4 are each independently selected from hydrogen or halogen; R.sub.5 is selected from the group consisting of ##STR00030##

2. The sulfanilamide derivative according to claim 1, wherein the sulfanilamide derivative is selected from the group consisting of ##STR00031## ##STR00032## ##STR00033##

3. The sulfanilamide derivative according to claim 1, wherein the sulfanilamide derivative is prepared as an anti-hepatitis B drug.

4. The sulfanilamide derivative according to claim 2, wherein the sulfanilamide derivative is prepared as an anti-hepatitis B drug.

Description

SPECIFIC EMBODIMENTS

(1) The following examples facilitate the understanding of the invention, but do not limit the content of the invention, and in the following examples, all the target compounds are numbered as above.

(2) Synthetic Route:

(3) ##STR00007##

(4) Reagents and conditions: (i) chlorosulfonic acid, 0° C., 6-12 h, 140-150° C.; (ii) chlorosulfoxide, N, N-dimethylformamide, 3-5h, 80° C.; (iii) acetonitrile, different types of aniline, 8 h, 60° C.; (iv) dichloromethane, N, N-disopropylethylamine, different types of amines, 8 h, 45° C.

Example 1: Preparation of Compound 2

(5) Take a 50 mL round bottom flask, add chlorosulfonic acid (18 mL, 277.54 mmol) and cool to 0° C. Slowly add 1, 4-benzodioxole-6-carboxylic acid (5.0032 g, 27.75 mmol) at low temperature, bring to room temperature, reflux at 100° C. for 6h. After reaction, cool to room temperature, add to 150 mL chilled water drop by drop, filtered, washed with water and dry to obtain 7.0040 g of brown solid in 90.6% yield.

(6) ##STR00008##

(7) NMR and MS Data of Compound 2

(8) .sup.1H-NMR (400 MHz, DMSO) δ 12.54 (s, 4H), 7.52 (d, J=1.6 Hz, 1H), 7.18 (d, J=2.0 Hz, 1H), 4.31 (d, J=4.0 Hz, 2H), 4.28 (d, J=4.8 Hz, 2H).

(9) .sup.13C-NMR (100 MHz, DMSO) δ166.63, 144.15, 143.44, 140.33, 120.71, 120.59, 117.91, 64.73, 64.12.

(10) ESI-MS: calculated for C.sub.9H.sub.7ClO.sub.6S [M−H].sup.− 277.96519, found 276.95691.

Example 2: Preparation of Compound 3

(11) Take a 25 ml round bottom flask, dissolve intermediate 2 (1 g, 3.58 mmol) in 10 ml sulfoxide chloride, add 2 drops of N, N-dimethylformamide, reflux reaction at 80° C., cool to room temperature after reaction, spin evaporation to obtain 1.17 g of yellow oil product.

Example 3: Preparation of Compound I

(12) Take a 100 mL round bottom flask, dissolve intermediate 3 in 20 mL acetonitrile, add different types of aniline, reflux reaction at 60° C., cool to room temperature after reaction, concentrate, add sample by dry method, fast prepare silica gel column for separation, dichloromethane-n-hexane solvent mixture recrystallization.

(13) ##STR00009##

Example 4: Preparation of Compound 5a

(14) Take a 100 ml round bottom flask, dissolve intermediate 3 (1 g, 3.37 mmol) in 20 mL acetonitrile, add 3-chloro-4-fluoroaniline (0.5886 g, 3.37 mmol), reflux reaction at 60° C., cool to room temperature after reaction, concentrate and add sample by dry method, fast preparation silica gel column separation, dichloromethane-n-hexane solvent mixture recrystallization, 1.0969 g yellow solid was obtained in 91% yield.

(15) ##STR00010##

(16) NMR and MS Data of Compound 5a

(17) .sup.1H-NMR (400 MHz, DMSO) δ 10.32 (s, 1H), 8.14-7.93 (m, 1H), 7.79-7.58 (m, 1H), 7.38 (dt, J=8.8, 1.2 Hz, 2H), 7.17 (d, J=2.0 Hz, 1H), 4.33 (dd, J=27.6, 3.1 Hz, 4H).

(18) .sup.13C-NMR (100 MHz, DMSO) δ 164.24 (d, J=11 Hz), 153.97 (d, J=201 Hz), 143.22, 141.98, 141.18, 136.54 (dd, J=11.3 Hz), 124.48 (d, J=9 Hz), 121.58 (d, J=9 Hz), 120.53 (t, J=8 Hz), 119.16, 117.44, 117.22, 116.94 (d, J=3 Hz), 65.10, 64.26.

(19) ESI-MS: calculated for C.sub.15H.sub.10C.sub.12FNO.sub.5S[M−H].sup.− 404.96408, found 403.95554.

Example 5: Preparation of Compound 5b

(20) Take a 25 mL round bottom flask, dissolve intermediate 5a (720 mg, 1.7 mmol) in 10 ml dichloromethane, add cyclopentylamine (156 μL, 1.7 mmol) N, N-diisopropylethylamine (570 μL, 5.1 mmol), and reflux at 40° C. After combing the organic phases, wash with saturated salt water (20 mL×3), dry over anhydrous sodium sulfate, concentrate, add sample by dry method, separate on a silica gel column by rapid preparative chromatography and recrystallize in a dichloromethane-n-hexane solvent mixture to obtain 0.21 g white solid powder in 27% yield with a melting point of 176.6-178.1° C.

(21) ##STR00011##

(22) NMR and MS Data of Compound 5b

(23) .sup.1H-NMR (400 MHz, DMSO) δ 10.43 (s, 1H), 8.02 (dd, J=6.7, 2.2 Hz, 1H), 7.71-7.53 (m, 3H), 7.48-7.32 (m, 2H), 4.42 (dd, J=30.3, 3.0 Hz, 4H), 2.05-0.93 (m, 9H).

(24) .sup.13C-NMR (100 MHz, DMSO) δ 163.37, 153.93 (d, J=242 Hz), 144.75, 144.22, 136.35 (d, J=3 Hz), 134.24, 125.68, 121.70, 120.65 (d, J=7 Hz), 120.28, 119.77, 119.59, 117.48 (d, J=12 Hz), 65.37, 64.35, 54.94, 32.97, 23.28.

(25) ESI-MS: calculated for C.sub.20H.sub.20ClFN.sub.2O.sub.5S[M−H].sup.− 455.08215, found 454.07655.

Example 6: Preparation of Compound 5c

(26) Take a 25 mL round bottom flask, dissolve compound 5a (300 mg, 0.7 mmol) in 10 mL dichloromethane, add cyclohexylamine (70 μL, 0.7 mmol) N, N-diisopropylethylamine (330 μL, 2.1 mmol), reflux reaction at 40° C. After the reaction cooled to room temperature, remove dichloromethane by spin evaporation, add water (40 mL) and extract by using ethyl acetate three times (20 mL×3), combine the organic phases, wash with saturated salt water (20 mL×3), dry with anhydrous sodium sulfate, concentrate and dichloromethane-n-hexane solvent mixture is recrystallized to obtain 0.18 g white solid powder, yield 54%, melting point 167.8-168.8° C.

(27) ##STR00012##

(28) NMR and MS Data of Compound 5c

(29) .sup.1H-NMR (400 MHz, DMSO) δ 10.43 (s, 1H), 8.02 (dd, J=6.7, 2.0 Hz, 1H), 7.65 (d, J=6.8 Hz, 2H), 7.57 (d, J=1.6 Hz, 1H), 7.50-7.36 (m, 2H), 4.99-3.92 (m, 4H), 2.93 (s, 1H), 1.60 (d, J=7.8 Hz, 4H), 1.37-0.96 (m, 6H).

(30) .sup.13C-NMR (100 MHz, DMSO) δ 163.26, 153.68 (d, J=232 Hz), 144.66, 144.19, 136.36 (d, J=4 Hz), 135.05, 125.64, 120.64 (d, J=7 Hz), 120.00, 119.77, 119.59, 117.60, 117.32 (d, J=14 Hz), 65.35, 64.35, 52.61, 33.68, 25.33, 24.76.

(31) ESI-MS: calculated for C.sub.21H.sub.22ClFN.sub.2O.sub.5S [M−H].sup.− 468.09220, found 467.08401.

Example 7: Preparation of Compound 5d

(32) Same operation as example 5, except that cyclopentylamine was replaced with 4-hydroxypiperidine. White solid powder, yield 50%, melting point 174.6-181.3° C.

(33) ##STR00013##

(34) NMR and MS data of compound 5d

(35) .sup.1H-NMR (400 MHz, DMSO) δ 10.48 (s, 1H), 8.02 (dd, J=6.8, 2.5 Hz, 1H), 7.78-7.58 (m, 1H), 7.55-7.38 (m, 2H), 7.31 (d, J=2.1 Hz, 1H), 4.70 (d, J=3.7 Hz, 1H), 4.43 (dd, J=29.6, 3.0 Hz, 4H), 3.56 (dd, J=7.0, 3.5 Hz, 1H), 3.22-3.00 (m, 2H), 2.78 (t, J=8.1 Hz, 2H), 1.96-1.68 (m, 2H), 1.62-1.29 (m, 2H).

(36) .sup.13C-NMR (100 MHz, DMSO) δ 163.26, 144.92 (d, J=102 Hz), 136.36, 128.06, 126.15, 121.69, 120.95, 120.63 (d, J=7 Hz), 119.77, 119.59, 118.26, 117.61, 117.40, 65.39, 64.36, 64.05, 43.55, 33.28.

(37) ESI-MS: calculated for C.sub.20H.sub.20ClFN.sub.2O.sub.6S[M−H].sup.− 470.07146, found 469.06342.

Example 8: Preparation of Compound 5e

(38) The operation was the same as in example 5, except that cyclopentylamine was replaced with 4-amino-1,2,4-triazole, and the product was a brown solid in 18% yield with a melting point of 252.1-253.2° C.

(39) ##STR00014##

(40) NMR and MS Data of Compound 5e

(41) .sup.1H-NMR (400 MHz, DMSO) δ 10.44 (s, 1H), 8.00 (dd, J=6.8, 2.4 Hz, 1H), 7.66-7.59 (m, 1H), 7.55 (t, J=8.9 Hz, 1H), 7.50 (d, J=2.3 Hz, 1H), 7.47-7.38 (m, 3H), 7.30-6.88 (m, 1H), 4.48 (d, J=41.6 Hz, 4H).

(42) .sup.13C-NMR (100 MHz, DMSO) δ 162.57, 154.44 (d, J=243 Hz), 152.80, 147.18, 144.52, 136.12, (d, J=3 Hz), 133.96, 132.76, 129.80, 126.04, 122.15, 121.82, 120.72 (d, J=7 Hz), 119.79, 118.99, 117.34, 65.72, 65.34.

(43) ESI-MS: calculated for C.sub.17H.sub.13ClFN.sub.5O.sub.5S[M+H].sup.+ 453.03100, found 453.98334.

Example 9: Preparation of Compound 5f

(44) The operation was the same as example 5, except that cyclopentylamine was replaced with imidazole and the product was a white solid with a yield of 22% and a melting point of 220.9-222.8° C.

(45) ##STR00015##

(46) NMR and MS Data of Compound 5f

(47) .sup.1H-NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 8.41 (s, 1H), 7.99 (dd, J=6.8, 2.3 Hz, 1H), 7.88-7.81 (m, 2H), 7.77 (d, J=2.2 Hz, 1H), 7.66-7.56 (m, 1H), 7.43 (t, J=9.1 Hz, 1H), 7.13 (s, 1H), 4.42 (dd, J=28.5, 3.2 Hz, 4H).

(48) .sup.13C-NMR (100 MHz, DMSO) δ 162.76, 153.98 (d, J=242 Hz), 147.28, 144.92, 137.77, 136.21, (d, J=3 Hz), 131.67, 129.27, 127.01, 121.09, 120.55 (d, J=7 Hz), 119.83, 118.79, 117.93, 117.67, 117.45, 65.57, 64.36.

(49) ESI-MS: calculated for C.sub.18H.sub.13ClFN.sub.3O.sub.5S[M−H].sup.−437.02485, found 436.01538.

Example 10: Preparation of Compound 5g

(50) The operation was the same as example 5, except that the cyclopentylamine was replaced with morpholine group and the product was a white solid, yield 78.7%, melting point 249.5-252.2° C.

(51) ##STR00016##

(52) NMR and MS Data of Compound 5g.

(53) .sup.1H-NMR (400 MHz, DMSO) δ 10.39 (s, 1H), 10.14 (s, 1H), 7.99 (dd, J=6.8, 2.3 Hz, 1H), 7.66-7.57 (m, 1H), 7.53 (d, J=2.0 Hz, 1H), 7.42 (t, J=9.1 Hz, 1H), 7.32 (d, J=2.0 Hz, 1H), 7.03 (q, J=8.4 Hz, 4H), 4.38 (dd, J=32.7, 2.9 Hz, 4H), 2.19 (s, 3H).

(54) .sup.13C-NMR (100 MHz, DMSO) δ 163.22, 154.16 (d, J=253 Hz), 145.73, 144.51, 137.77, 136.35 (d, J=2.9 Hz), 126.78, 126.30, 121.69, 121.13, 120.63 (d, J=7.0 Hz), 119.68 (d, J=18.5 Hz), 118.45, 117.60, 117.39, 65.72, 65.4.

(55) ESI-MS: calculated for C.sub.19H.sub.18ClFN.sub.2O.sub.6S[M−H].sup.−456.05581, found 455.04691.

Example 11: Preparation of Compound 5h

(56) The operation was the same as example 5, except that cyclopentylamine was replaced with aniline and the product was a white solid with a yield of 41% and a melting point of 223.0-226.8° C.

(57) ##STR00017##

(58) NMR and MS Data of Compound 5h

(59) .sup.1H-NMR (400 MHz, DMSO) δ 10.41 (t, J=47.3 Hz, 2H), 7.98 (d, J=6.8 Hz, 1H), 7.69-7.49 (m, 2H), 7.45-7.31 (m, 3H), 7.30-7.20 (m, 1H), 7.19-7.09 (m, 1H), 7.03 (t, J=7.3 Hz, 1H), 4.38 (d, J=33.2 Hz, 4H).

(60) .sup.13C-NMR (100 MHz, DMSO) δ 163.04 (d, J=9.9 Hz), 154.53 (d, J=242.1 Hz), 136.25, 131.58, 129.76, 125.86 (d, J=10.5 Hz), 124.54, 122.02, 121.68 (d, J=5.2 Hz), 120.98-120.08, 119.70 (d, J=18.5 Hz), 118.02, 117.50 (dd, J=14.9, 6.8 Hz), 65.41, 64.31.

(61) ESI-MS: calculated for C.sub.21H.sub.16ClFN.sub.2O.sub.5S[M−H].sup.−462.04525, found 461.03491.

Example 12: Preparation of Compound 5i

(62) The operation was the same as example 5, except that cyclopentylamine was replaced with p-methylaniline and the product was a white solid with a yield of 70% and a melting point of 229.1-237.0° C.

(63) ##STR00018##

(64) NMR and MS Data of Compound 5i

(65) .sup.1H-NMR (400 MHz, DMSO) δ 10.39 (s, 1H), 10.14 (s, 1H), 7.99 (dd, J=6.8, 2.3 Hz, 1H), 7.66-7.57 (m, 1H), 7.53 (d, J=2.0 Hz, 1H), 7.42 (t, J=9.1 Hz, 1H), 7.32 (d, J=2.0 Hz, 1H), 7.03 (q, J=8.4 Hz, 4H), 4.38 (dd, J=32.7, 2.9 Hz, 4H), 2.19 (s, 3H).

(66) .sup.13C-NMR (100 MHz, DMSO) δ 163.11, 153.95 (d, J=242 Hz), 145.24, 144.20, 136.25, 135.41, 133.83, 132.29, 130.16, 125.73, 121.68, 120.70, 120.59, 120.51, 119.70 (d, J=18.3 Hz), 117.58 (d, J=18.3 Hz), 117.38, 65.36, 64.30, 20.75.

(67) ESI-MS: calculated for C.sub.22H.sub.18ClFN.sub.2O.sub.5S[M−H].sup.−476.06090, found 475.05130.

Example 13: Preparation of Compound 5j

(68) The product was prepared as in example 5, except that cyclopentylamine was replaced with 4-butylaniline and the product was a white solid with a yield of 75% and a melting point of 168.9-171.9° C.

(69) ##STR00019##

(70) NMR and MS Data of Compound 5j

(71) .sup.1H-NMR (400 MHz, DMSO) δ 10.38 (s, 1H), 10.14 (s, 1H), 7.99 (d, J=4.8 Hz, 1H), 7.64-7.57 (m, 1H), 7.51 (s, 1H), 7.41 (t, J=9.1 Hz, 1H), 7.32 (d, J=1.3 Hz, 1H), 7.05 (dd, J=19.2, 8.3 Hz, 4H), 4.59-4.13 (m, 4H), 2.46 (t, J=7.6 Hz, 2H), 1.58-1.37 (m, 2H), 1.24 (dd, J=14.6, 7.3 Hz, 2H), 0.84 (t, J=7.3 Hz, 3H).

(72) .sup.13C-NMR (101 MHz, DMSO) δ 163.10, 153.94 (d, J=242 Hz), 145.23, 144.20, 138.78, 136.29, 135.59, 132.37, 129.47, 125.73, 121.66, 120.73, 120.6, 120.56, 120.43, 119.69 (d, J=18.4 Hz), 117.5 (d, J=6 Hz), 117.37, 65.36, 64.30, 34.54, 33.51, 22.15, 14.19.

(73) ESI-MS: calculated for C.sub.26H.sub.26ClFN.sub.2O.sub.5S[M−H].sup.−518.10785, found 517.09808.

Example 14: Preparation of Compound 5k

(74) The product was prepared as in example 5, except that cyclopentylamine was replaced with isopropylamine and the product was a white solid with a yield of 66% and a melting point of 170.4-172.3° C.

(75) ##STR00020##

(76) NMR and MS Data of Compound 5k

(77) .sup.1H-NMR (400 MHz, DMSO) δ 10.43 (s, 1H), 8.02 (dd, J=6.8, 2.4 Hz, 1H), 7.76-7.51 (m, 3H), 7.51-7.34 (m, 2H), 4.42 (dd, J=29.8, 3.1 Hz, 4H), 3.23 (dd, J=13.1, 6.6 Hz, 1H), 0.98 (d, J=6.5 Hz, 6H).

(78) .sup.13C-NMR (100 MHz, DMSO) δ 163.37, 154.23 (d, J=252.4 Hz), 144.71, 144.23, 135.37, 134.55, 125.71, 121.69, 120.65 (d, J=7 Hz), 120.12, 119.77, 117.61, 117.39, 111.69, 65.35, 64.36, 46.77, 23.72.

(79) ESI-MS: calculated for C.sub.18H.sub.18ClFN.sub.2O.sub.5S[M−H].sup.−428.06090, found 427.05164.

Example 15: Preparation of Compound 51

(80) The product was prepared as in example 5, except that cyclopentylamine was replaced with 2-ethylhexylamine and the product was a white solid with a yield of 40% and a melting point of 190.9-193.7° C.

(81) ##STR00021##

(82) NMR and MS Data of Compound 51

(83) .sup.1H-NMR (400 MHz, DMSO) δ 10.43 (s, 1H), 8.01 (d, J=5.5 Hz, 1H), 7.76-7.25 (m, 5H), 4.41 (d, J=28.9 Hz, 4H), 2.63 (s, 2H), 1.46-1.04 (m, 9H), 0.96-0.63 (m, 6H).

(84) .sup.13C-NMR (100 MHz, DMSO) δ 163.37, 153.98 (d, J=246 Hz), 144.78, 144.27, 136.30, 133.37, 125.71, 121.71, 120.64, 120.21, 119.71, 119.63, 117.60, 117.43, 65.37, 64.36, 45.81, 30.62, 28.62, 23.74, 22.87, 14.39, 10.99.

(85) ESI-MS: calculated for C.sub.23H.sub.28ClFN.sub.2O.sub.5S[M−H].sup.−498.13915, found 497.12875.

Example 16: Preparation of Compound 5m

(86) The product was prepared as in example 5, except that p-hydroxycyclohexylamine was used instead of cyclopentylamine and the product was a yellow solid with a yield of 58% and a melting point of 201.1-202.4° C.

(87) ##STR00022##

(88) NMR and MS Data of Compound 5m

(89) .sup.1H-NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 8.01 (dd, J=6.8, 2.4 Hz, 1H), 7.62 (m, 3H), 7.41 (dd, J=12.3, 9.2 Hz, 2H), 4.73-3.81 (m, 5H), 3.30 (d, J=9.2 Hz, 1H), 3.13-2.72 (m, 1H), 1.67 (dd, J=32.9, 11.1 Hz, 4H), 1.39-0.87 (m, 4H).

(90) .sup.13C-NMR (100 MHz, DMSO) δ 163.36, 154.24 (d, J=242 Hz), 144.69, 144.21, 136.33 (d, J=3.0 Hz), 134.86, 125.66, 121.71, 120.66 (d, J=7 Hz)), 120.02, 119.78, 119.60, 117.37 (t, J=21 Hz), 68.02, 65.35, 64.34, 52.14, 33.98, 31.44.

(91) ESI-MS: calculated for C.sub.21H.sub.22ClFN.sub.2O.sub.6S[M−H].sup.−484.08711, found 483.07886.

Example 17: Preparation of Compound 5n

(92) The first operation was the same as example 4, replacing 3-chloro-4-fluoroaniline with 3-fluoroaniline, and the subsequent operation was the same as example 16, with the product being a white solid in 40% yield and a melting point of 111.1-120.1° C.

(93) ##STR00023##

(94) NMR and MS Data of Compound 5n

(95) .sup.1H-NMR (400 MHz, DMSO) δ 10.43 (s, 1H) 7.70 (d, J=11.6 Hz, 1H), 7.62 (d, J=7.0 Hz, 1H), 7.56 (d, J=2.0 Hz, 1H), 7.52-7.35 (m, 3H), 6.95 (td, J=8.5, 2.2 Hz, 1H), 4.66-4.14 (m, 5H), 3.33-3.26 (m, 1H), 3.05-2.76 (m, 1H), 1.68 (dd, J=31.9, 11.5 Hz, 4H), 1.30-0.90 (m, 4H).

(96) .sup.13C-NMR (100 MHz, DMSO) δ 163.79, 162.44 (d, J=209 Hz), 144.66, 144.20, 140.81 (d, J=11.0 Hz), 134.85, 130.94 (d, J=9.5 Hz), 125.95, 119.96, 117.17, 116.02, 110.85 (d, J=21 Hz), 106.99 (d, J=27 Hz), 68.02, 65.33, 64.35, 55.37, 52.14, 33.98, 31.44.

(97) ESI-MS: calculated for C.sub.21H.sub.23FN.sub.2O.sub.6S[M−H].sup.−450.12609, found 449.11536.

Example 18: Preparation of Compound 5o

(98) The first operation was the same as example 4, replacing 3-chloro-4-fluoroaniline with p-fluoroaniline, the subsequent operation was the same as example 16, the product was a white solid with a yield of 42% and a melting point of 205.1-207.5° C.

(99) ##STR00024##

(100) NMR and MS Data of Compound 5o

(101) .sup.1H-NMR (400 MHz, DMSO) δ 10.30 (s, 1H), 7.73 (dd, J=8.1, 5.3 Hz, 2H), 7.63 (d, J=6.9 Hz, 1H), 7.55 (s, 1H), 7.38 (s, 1H), 7.21 (t, J=8.7 Hz, 2H), 4.45 (dd, J=26.2, 23.1 Hz, 5H), 3.29 (s, 1H), 2.94-2.80 (m, 1H), 1.67 (dd, J=33.1, 11.4 Hz, 4H), 1.35-0.75 (m, 4H).

(102) .sup.13C-NMR (100 MHz, DMSO) δ 163.07, 159.27 (d, J=333 Hz), 144.62, 144.16, 135.71, 134.76, 126.13, 122.04 (d, J=8 Hz), 119.97, 117.03, 115.99, 115.76, 68.01, 66.29, 64.39, 52.12, 33.98, 31.44.

(103) ESI-MS: calculated for C.sub.21H.sub.23FN.sub.2O.sub.6S[M−H].sup.−450.12609, found 449.11542.

Example 19: Preparation of Compound 5p

(104) The first operation was the same as example 4, replacing 3-chloro-4-fluoroaniline with o-fluoroaniline, and the subsequent operation was the same as example 16. The product was a white solid with a yield of 45% and a melting point of 200.0-200.8° C.

(105) ##STR00025##

(106) NMR and MS data of compound 5p

(107) .sup.1H-NMR (400 MHz, DMSO) δ 10.06 (s, 1H), 8.04 (t, J=8.6 Hz, 1H), 7.79 (d, J=1.6 Hz, 1H), 7.65 (d, J=7.0 Hz, 1H), 7.43 (d, J=1.7 Hz, 1H), 7.38-7.28 (m, 1H), 7.27-7.17 (m, 2H), 4.47 (d, J=51.4 Hz, 5H), 3.31-3.25 (m, 1H), 2.97-2.82 (m, 1H), 1.68 (dd, J=33.1, 11.5 Hz, 4H), 1.13 (ddd, J=35.5, 17.0, 8.7 Hz, 4H).

(108) .sup.13C-NMR (100 MHz, DMSO) δ 162.57, 154.23 (d, J=241 Hz), 145.29, 144.37, 134.93, 126.25 (d, J=11.1 Hz), 125.03, 124.61, 123.87, 120.91, 117.87, 115.97 (d, J=19.2 Hz), 68.03, 65.67, 64.21, 52.17, 34.00, 31.44.

(109) ESI-MS: calculated for C.sub.21H.sub.23FN.sub.2O.sub.6S[M−H].sup.−450.12609, found 449.12653.

Example 20: Preparation of Compound 5q

(110) The first operation was the same as example 4, replacing 3-chloro-4-fluoroaniline with o-fluoroaniline, and the subsequent operation was the same as example 16. The product was a white solid with a yield of 45% and a melting point of 200.0-200.8° C.

(111) ##STR00026##

(112) NMR and MS Data of Compound 5q

(113) .sup.1H-NMR (400 MHz, DMSO) δ 10.59 (s, 1H), 7.63 (d, J=7.0 Hz, 1H), 7.56 (d, J=2.1 Hz, 1H), 7.46 (d, J=7.7 Hz, 2H), 7.41 (d, J=2.1 Hz, 1H), 6.98 (t, J=5.7 Hz, 1H), 4.42 (dd, J=29.4, 3.3 Hz, 5H), 3.31-3.25 (m, 1H), 3.00-2.81 (m, 1H), 1.68 (dd, J=32.7, 12.0 Hz, 4H), 1.26-0.98 (m, 4H).

(114) .sup.13C-NMR (100 MHz, DMSO) δ 163.77, 162.77 (d, J=199 Hz), 144.69, 144.23, 134.92, 125.58, 119.95, 117.36, 103.25, 102.96, 99.53, 68.01, 65.37, 64.36, 52.14, 33.97, 31.43.

(115) ESI-MS: calculated for C.sub.21H.sub.22F.sub.2N.sub.2O.sub.6S[M−H].sup.−468.11666, found 467.10590.

Example 21: Preparation of Compound 5r

(116) The first operation was the same as example 4, replacing 3-chloro-4-fluoroaniline with 3,4,5-trifluoroaniline, and the subsequent operation was the same as example 16, the product was a white solid with a yield of 50% and a melting point of 225.1-228.0° C.

(117) ##STR00027##

(118) NMR and MS data of compound 5r

(119) .sup.1H-NMR (400 MHz, DMSO) δ 10.57 (s, 1H), 7.69-7.60 (m, 3H), 7.56 (d, J=1.9 Hz, 1H), 7.41 (d, J=1.9 Hz, 1H), 4.64-4.13 (m, 5H), 3.30 (d, J=4.2 Hz, 1H), 2.89 (s, 1H), 1.67 (dd, J=34.3, 11.6 Hz, 4H), 1.43-0.91 (m, 4H).

(120) .sup.13C-NMR (100 MHz, DMSO) δ 163.70, 150.65 (d, J=202 Hz), 144.69, 144.24, 134.94, 125.38, 119.95, 117.41, 104.74, 104.50, 68.01, 65.38, 64.36, 52.14, 33.97, 31.43.

(121) ESI-MS: calculated for C.sub.20H.sub.21F.sub.3N.sub.2O.sub.3S [M−H].sup.−486.10724, found 485.09830.

Example 22: Preparation of Compound 5s

(122) The first operation was the same as example 4, replacing 3-chloro-4-fluoroaniline with 3-fluoro-4-methylaniline, and the subsequent operation was the same as example 16, the product was a white solid with a yield of 42% and a melting point of 201.1-202.4° C.

(123) ##STR00028##

(124) NMR and MS Data of Compound 5s

(125) .sup.1H-NMR (400 MHz, DMSO) δ 10.18 (s, 1H), 7.62 (d, J=6.9 Hz, 2H), 7.58-7.50 (m, 2H), 7.38 (d, J=1.7 Hz, 1H), 4.41 (dd, J=31.0, 3.2 Hz, 5H), 3.29 (dd, J=9.0, 5.0 Hz, 1H), 3.00-2.75 (m, 1H), 2.24 (s, 3H), 1.68 (dd, J=31.9, 11.7 Hz, 4H), 1.25-0.94 (m, 5H).

(126) .sup.13C-NMR (100 MHz, DMSO) δ 162.96, 157.43 (d, J=238 Hz), 144.65, 144.13, 135.15, 134.78, 126.07, 124.80 (d, J=18 Hz), 123.29 (d, J=5 Hz), 120.02, 119.53 (d, J=8 Hz), 117.05, 115.57, 115.34, 68.02, 65.32, 64.33, 52.14, 33.99, 31.43, 14.83.

(127) ESI-MS: calculated for C.sub.22H.sub.25FN.sub.2O.sub.6S[M−H].sup.−464.14174, found 463.13527.

Example 23: In Vitro Anti-HBV Cellular Activity Screening Assay of Target Compounds Test Principle

(128) HBV stably transfected hepatocellular carcinoma cell line, HepAD38, was able to secrete HBV viral particles (containing HBsAg, HBeAg and DNA) when the cells were cultured. The amount of HBsAg and HBeAg secreted by the cells and the DNA produced varied under the intervention of anti-HBV target compounds, so the antiviral activity of the sample drug could be calculated by measuring the amount of HBsAg and HBeAg secreted by the cells into the culture supernatant and the HBV DNA produced, with reference to the amount in the unspiked control. Using entecavir as a positive control drug, the concentration of the drug at 50% inhibition of HBV DNA was measured by quantitative PCR at the value of IC.sub.50; the concentration of the sample drug causing 50% cell death was measured by CCK-8 at the value of CC.sub.50, and the “selectivity index” of the compound to be tested was calculated. SI=CC.sub.50/IC.sub.50. (1) Cytotoxicity assay

(129) Each sample was prepared with HepAD38 cell culture medium at a diluted concentration (1 μM) for preliminary activity screening, and a blank control was set up and a positive control drug was added to the 96-well cell culture plate with 3 replicate wells per concentration, and the same concentration was changed every 2 days and a drug-free cell control group was set up for 10 days. Cell viability was measured by CCK-8 assay to determine the toxicity of the drug to HepAD38 cells. Five dilutions (10 μM, 2 μM, 0.4 μM, 0.08 μM and 0.012 μM/L) were prepared in HepAD38 cell culture medium for compounds with low toxicity, and a blank control and entecavir and lead compound 5a were used as positive control drugs. Cell culture plates were added to 96-well plates with 3 replicate wells per concentration, and the same concentration was changed every 2 days and a drug-free cell control group was set up for 10 days. Cell viability was measured by CCK-8 assay to determine the toxicity of the drug on HepAD38 cells.

(130) (2) HBV DNA Activity Inhibition Assay (Quantitative PCR Method)

(131) HepAD38 cells were cultured in 96-well cell culture plate for 24 hours, then 10 μM, 2 μM, 0.4 μM, 0.08 μM and 0.012 μM drug-containing medium was added and the cells were incubated for 10 days (changed every 2 days), the supernatant was collected and the supernatant HBV DNA content was measured by quantitative PCR.

(132) TABLE-US-00001 TABLE 1 Anti-hepatitis B virus activity of the active compounds and the marketed drug entecavir CC.sub.50 EC.sub.50 IC.sub.90 Number ((μM) (μM) (μM) SI 5b >10 0.00252 0.293 >100 5c >10 0.01490 0.430 >100 5k >10 0.00096 0.099 >100 ETV >10 0.000121 0.021 >100

(133) As shown in Table 1, further in vitro evaluation of the anti-HBV activity of initially screened target compounds 5b, 5c and 5k was performed based on the results of the initial screening, and the cytotoxicity of the drugs at different concentrations was determined by CCK-8 assay; the activity of the drugs in inhibiting HBV DNA at different concentrations was determined by quantitative PCR. Five concentration gradients (10 μM, 2 μM, 0.4 μM, 0.08 μM and 0.012 μM) were set for each compound with the listed drug entecavir as positive control, and the half inhibition concentration CC.sub.50, IC.sub.50 and selectivity coefficient SI were calculated, respectively.

(134) The activity results showed that the target compounds 5b, 5c, and 5k all exhibited less cytotoxicity with CC.sub.50>10 μM and 5k showed better anti-HBV DNA replication activity. Its IC.sub.50 was 0.00096 μM, which is not as good as the marketed drug entecavir, but still allows for further modification studies.