Application of five-membered sulfur-containing heterocyclic nucleoside lead compound in anti-Zika virus active medicine

Abstract

The invention discloses application of a five-membered sulfur-containing heterocyclic nucleoside lead compound in anti-Zika virus active drugs, and belongs to the technical field of medicinal chemistry. A series of five-membered sulfur-containing heterocyclic ring nucleoside compounds are synthesized through a Sulfa-Michael / Aldol cascade reaction or a Michael addition reaction. Results show that in an anti-Zika virus test, the five-membered sulfur-containing heterocyclic nucleoside lead compounds 39b, 22r and 22n have relatively good anti-Zika virus activity, and a new structural unit is provided for anti-virus candidate drug molecules.

Description

Technical Field

[0001] The invention relates to application of a five-membered sulfur-containing heterocyclic nucleoside lead compound in an anti-Zika virus active drug, and belongs to the field of medicinal chemistry. Background Art

[0002] Viruses seriously threaten human life and health and are one of the main causes of human death. The research and development of antiviral drugs has always been one of the important directions in drug development. Artificially synthesized nucleoside compounds are similar in structure to natural nucleoside compounds. After entering the human body, they can inhibit the replication of viruses or tumor cells, achieving the purpose of antiviral or anti-tumor.

[0003] People pay more and more attention to the research and development of nucleoside anti-tumor and antiviral drugs. Five-membered sulfur heterocyclic nucleoside compounds are a unique class of compounds among nucleoside compounds, with unique physical and chemical properties, and have become the focus of nucleoside drug research and development.

[0004] Currently, five-membered sulfur heterocyclic nucleoside compounds are an important class of antiviral drugs on the market, but there are relatively few studies on the antiviral activity of five-membered sulfur heterocyclic nucleoside compounds. Innovative research on five-membered sulfur heterocyclic nucleoside antiviral drugs can not only break the international monopoly on nucleoside drugs, but also has important scientific significance and potential application prospects. Summary of the invention

[0005] In order to overcome the above technical defects, the present invention synthesizes a series of five-membered sulfur heterocyclic nucleoside compounds through reactions such as Sulfa-Michael / Aldol tandem reaction, provides nucleoside lead compounds with more functional structures, and discloses five-membered sulfur heterocyclic nucleoside lead compounds with anti-Zika virus activity.

[0006] The five-membered sulfur heterocyclic nucleoside lead compound of the present invention includes its racemate and chiral compound, and its general structural formula is as follows:

[0007]

[0008] Where: R 1 is selected from hydrogen, halogen, amino, C1-C20 alkyl, C1-C20 alkoxy; R 2 is selected from hydrogen, halogen, C1-C20 alkyl, C1-C20 alkoxy, C1-20 alkylthio, C2-C20 alkynylthio, halogenated C1-C20 alkylthio, benzylthio, phenylthio, substituted phenylthio, di(C1-C8 alkyl)amine, X=CH 2 , O, S, NH, N-(C1-C4)alkyl; n=0-5.

[0009] Furthermore, under the preferred conditions, R1 is selected from hydrogen, halogen, amino, C1-C20 alkyl, C1-C20 alkoxy; R 2 is selected from hydrogen, halogen, C1-C20 alkyl, C1-C20 alkoxy, C1-20 alkylthio, C2-C20 alkynylthio, halogenated C1-C20 alkylthio, benzylthio, phenylthio, substituted phenylthio, di(C1-C8 alkyl)amine, X=CH 2 , O, S, NH, N-(C1-C4)alkyl; n = 0-5; R 3 =C1-C8 alkyl, halogen, trifluoromethyl, benzyl, substituted phenylmethyl, morpholinyl; wherein the substituent in the substituted phenyl is halogen or C1-C8 alkyl; R is selected from ethyl.

[0010] Furthermore, under the preferred conditions, R 1 is selected from hydrogen, halogen, amino; R 2 is selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-6 alkylthio, C2-C4 alkynylthio, halogenated C1-C4 alkylthio, benzylthio, phenylthio, substituted phenylthio, di(C1-C2 alkyl)amine, X=CH 2 , O, S, NH, N-(C1-C4)alkyl; n = 1-3; R 3 =C1-C4 alkyl, halogen, trifluoromethyl, benzyl, substituted phenylmethyl, morpholinyl; wherein the substituent in the substituted phenyl is halogen or C1-C4 alkyl; R is selected from ethyl.

[0011] Further, under the most preferred conditions, the structure of the five-membered sulfur heterocyclic nucleoside lead compound is

[0012]

[0013] The present invention also provides the use of the five-membered sulfur heterocyclic nucleoside lead compound and a pharmaceutically acceptable salt thereof in the preparation of antiviral drugs.

[0014] Furthermore, in the above technical solution, the antiviral agent is anti-Zika virus.

[0015] Furthermore, in the above technical solution, the pharmaceutically acceptable salt includes a salt formed by an acyclic nucleoside compound and an organic acid or an inorganic acid.

[0016] Furthermore, in the above technical solution, the organic acid is selected from one or more of malic acid, lactic acid, camphorsulfonic acid, citric acid, fumaric acid or oxalic acid, and the organic acid is selected from one or more of phosphoric acid, hydrohalic acid, sulfuric acid or nitric acid.

[0017] The present invention further provides a pharmaceutical composition containing the above-mentioned general structural formula or specific compound.

[0018] Beneficial effects of the invention:

[0019] 1. The raw materials for the preparation of the five-membered sulfur heterocyclic nucleoside lead compound are simple and easy to obtain, and have broad application prospects.

[0020] 2. The products were tested for antiviral activity, and it was found that compounds 39b, 22r and 22n had good anti-Zika virus activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a flow chart of the research and development method in Example 3;

[0022] Figure 2 The anti-Zika virus activity graph of 39b in Example 3;

[0023] Figure 3 The anti-Zika virus activity graph of 22r in Example 3;

[0024] Figure 4 This is a graph showing the anti-Zika virus activity of 22n in Example 3. DETAILED DESCRIPTION

[0025] Example 1

[0026]

[0027] Typical operation of the first step: In a round-bottom flask, purine compound A (10 mmol), triphenylphosphine (0.0521 g, 0.2 mL), sodium acetate (0.1641 g, 2 mmol), acetic acid (0.3 mL, 2 mmol) and ethyl propiolate (1.21 mL, 12 mmol) were added to toluene (100 mL). Under nitrogen protection, the mixture was heated and stirred at 110 ° C for 12 h, and dichloromethane was added to extract the reaction mixture 3 times; the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by silica gel (PE / EA=2 / 1) column chromatography to obtain product B.

[0028] Typical operation of the second step: α-heterocyclic acrylate B (0.05 mmol) and 1,4-disulfane 2,5-diol (0.03 mmol) were added to a round-bottom flask. Then, 20 mmol% triethylamine was added and stirred at room temperature for 4 hours. The reaction product was detected by thin layer chromatography, and the solvent was removed by silica gel (PE / EA=2 / 1) column chromatography to obtain products a and b.

[0029] Structural characterization of representative compounds:

[0030] 1.2.1.(±)-Ethyl-4-hydroxy-3-(9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(1a,1b)

[0031] 1a:white solids,41.2%yield. 1 H NMR(600MHz,CDCl 3 )δ9.18(s,1H),8.95(s,1H),8.42(s,1H),5.42(d,J=42.6Hz,2H),4.24-4.16(m,2H),3.96(d,J=12.6Hz,1H),3.59(d,J=12Hz,1H),3.19(dd,J=4.8Hz,J=7.2Hz,1H),2.86(dd,J=4.8Hz,J=11.4Hz,1H),1.08(t,J=6.6Hz,3H). 13 C NMR(150MHz,CDCl 3 )δ169.3,152.4,151.5,149.6,144.0,134.3,78.9,74.0,63.3,35.5,35.4,13.9.ESI-HRMS(m / z)calcd C 12 H 15 N 4 O 3 S(M+H) + ,295.0859;found,295.0856.HPLC purity:97.7%.1b:white solids,52.4%yield. 1 H NMR(600MHz,CDCl 3 )δ9.06(s,1H),8.83(s,1H),8.51(s,1H),5.49(t,J=6.0 Hz,1H),4.22-4.15(m,2H),3.74(dd,J=12 Hz,J=30.0Hz,2H),3.34(q,J=5.4 Hz,1H),2.89(q,J=6,1H),1.12(t,J=7.2 Hz,3H), 13 CNMR(150 MHz,CDCl 3 )δ168.1,152.1,151.7,148.9,145.0,133.7,76.3,73.8,63.1,34.7,34.0,13.8;ESI-HRMS(m / z)calcd C 12 H 14 N4 O 3 SNa(M+Na) + ,317.0679;found,317.0678.HPLC purity:98.0%.

[0032] 1.2.5.(±)-Ethyl-4-hydroxy-3-(6-(piperidin-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(5a,5b)

[0033] 5a:white solid,41.7%yield. 1 H NMR(400 MHz,CDCl 3 )δ8.24(s,1H),7.90(s,1H),5.25(t,J=5.6 Hz,1H),4.28-4.15(m,6H),3.87(d,J=12.0 Hz,1H),3.53(d,J=12.0Hz,1H),3.25(dd,J=6.0,J=11.2 Hz,1H),2.94(dd,J=11.2,J=5.2 Hz,1H),1.76-1.69(m,6H),1.15(t,J=7.2 Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ169.8,154.1,151.9,150.9,136.8,119.8,79.7,74.0,63.0,35.9,35.1,26.3,24.9,14.1.ESI-HRMS(m / z)calcdC 17 H 24 N 5 O 3 S(M+H) + ,378.1594;found,378.1597.HPLC purity:98.7%.5b:white solid,51.1%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.19(s,1H),7.99(s,1H),5.39(t,J=6.6,1H),4.24-4.18(m,6H),3.72(d,J=12,1H),3.44(d,J=12,1H),3.21(q,J=6,1H),2.80(q,J=7.2,1H),1.74-1.67(m,6H),1.18(t,J=6.6,3H). 13C NMR(150 MHz,CDCl 3 )δ168.6,154.1,151.9,150.7,137.5,119.6,73.8,63.0,35.2,33.4,26.3,24.8,14.0.ESI-HRMS(m / z)calcdC 17 H 23 N 5 O 3 SNa(M+Na) + ,400.1414;found,400.1410.HPLC purity:95.8%.

[0034] 1.2.9.(±)-Ethyl-4-hydroxy-3-(6-morpholino-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(9a,9b)

[0035] 9a:white solid,24.4%yield. 1 H NMR(400 MHz,CDCl 3 )δ8.27(s,1H),7.94(s,1H),5.26(t,J=5.6 Hz,1H),4.31(s,4H),4.26-4.15(m,2H),3.88(d,J=12.4 Hz,1H),3.83(d,J=4.0 Hz,4H),3.53(d,J=12.0 Hz,1H),3.23(q,J=6.0 Hz,1H),2.92(dd,J=5.2 Hz,J=11.6 Hz,1H),1.15(t,J=6.8 Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ169.7,154.1,151.9,151.1,137.3.120.0,79.6,74.0,67.1,63.0,35.8,35.2,14.1.ESI-HRMS(m / z)calcdC 16 H 22 N 5 O 4 S(M+H) + ,380.1387;found,380.1390.HPLC purity:97.9%.9b:white solid,45.4%yield. 1 H NMR(400 MHz,CDCl 3)δ8.23(s,1H),8.02(s,1H),5.94(s,1H),5.40(t,J=6.4 Hz,1H),4.29(s,4H),4.21(q,J=7.2 Hz,2H),3.81(t,J=4.8 Hz,4H),3.73(d,J=11.6 Hz,1H),3.46(d,J=11.6 Hz,1H),3.23(dd,J=6.0 Hz,J=10.8 Hz,1H),2.80(dd,J=7.2 Hz,J=10.8 Hz,1H),1.18(t,J=6.8Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ168.5,154.1,151.8,150.9,138.1,119.8,77.3,73.8,67.1,63.0,35.2,33.4,14.0.ESI-HRMS(m / z)calcd C 16 H 21 N 5 O 4 SNa(M+Na) + ,402.1206;found,402.1196.HPLC purity:99.4%.

[0036] 1.2.12.(±)-Ethyl-3-(6-(4-fluoropiperidin-1-yl)-9H-purin-9-yl)-4-hydroxytetrahydrothiophene-3-carboxylate(12a,12b)

[0037] 12a:white solid,42.3%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.21(s,1H),7.90(s,1H),6.48(s,1H),5.21(t,J=6 Hz,1H),4.93-4.84(m,1H),4.46(s,2H)4.19-4.13(m,4H),3.83(d,J=12 Hz,1H),3.50(d,J=12.6 Hz,1H),3.18(q,J=6 Hz,1H),2.89(dd,J=4.8Hz,J=11.4 Hz,1H),1.99-1.90(m,4H),1.10(t,J=7.2 Hz,3H). 13 C NMR(150 MHz,CDCl 3)δ169.5,153.8,151.8,150.9,137.0,119.7,88.6,87.5,79.4,73.9,62.8,42.3,35.7,35.0,31.4(Jc-F=19.7),13.9. 19 F NMR(376MHz,CDCl 3 )δ-181.58.ESI-HRMS(m / z)calcdC 17 H 22 FN 5 O 3 SNa(M+Na) + ,396.1500;found,396.1506.HPLC purity:98.9%.12b:whitesolid,53.4%yield. 1 H NMR(400 MHz,CDCl 3 )δ8.24(s,1H),8.03(s,1H),6.10(s,1H),5.41(t.J=6.0 Hz,1H),5.03-4.86(m,1H),4,54(s,2H),4.22(q,J=7.2 Hz,4H),3.75(d,J=12Hz,1H),3.44(d,J=11.6 Hz,1H),3.23(dd,J=10.8 Hz,J=6.0 Hz,1H),2.81(dd,J=10.8Hz,J=7.2 Hz,1H),2.05-1.95(m,4H),1.20(t,J=7.2 Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ168.6,154.1,151.9,150.9,137.9,119.8,87.3,73.8,63.1,35.3,33.3,31.6(d,J C-F =20.2Hz),14.0. 19 F NMR(376MHz,CDCl 3 )δ-181.68.ESI-HRMS(m / z)calcd C 17 H 22 FN 5 O 3 SNa(M+Na) + ,396.1500;found,396.1508.HPLC purity:97.2%.

[0038] 1.2.15.(±)-Ethyl-4-hydroxy-3-(6-(4-morpholinopiperidin-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(15a,15b)

[0039] 15a:white solid,37.7%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.22(s,1H),8.02(s,1H),5.50(s,1H),5.40(t,J=9.6 Hz,1H),4.22(q,J=10.8 Hz,2H),3.76(s,1H),3.72(t,J=6.6 Hz,4H),3.44(d,J=18 Hz,1H),3.22(dd,J=9 Hz,J=16.2 Hz,1H),3.13(s,2H),2.80(dd,J=11.4 Hz,J=16.2 Hz,1H),2.59-2.52(m,5H),2.01(d,J=19.2 Hz,2H),1.62-1.52(m,4H),1.19(t,J=10.8 Hz,3H). 13 C NMR(150 MHz,CDCl 3 )δ169.6,154.0,150.8,137.8,119.8,73.8,67.4,63.1,62.2,49.9,35.3,33.3,28.6,14.0.ESI-HRMS(m / z)calcdC 21 H 31 N 6 O 4 S(M+H) + ,463.2111;found,463.2113.HPLC purity:97.9%.15b:white solid,38.6%yield. 1 H NMR(600 MHz,CDCl 3)δ8.25(s,1H),7.92(s,1H),6.55(s,1H)5.49(s,1H),5.25(t,J=8.4 Hz,1H),4.26-4.17(m,2H),3.88(d,J=18.6 Hz,1H),3.73(t,J=6.6 Hz,4H),3.53(d,J=18 Hz,1H),3.24(dd,J=9 Hz,J=16.8 Hz,1H),3.12(s,2H),2.93(q J=7.8 Hz,1H),2.60-2.53(s,5H),2.02(d,J=18 Hz,2H),1.63-1.52(m,4H),1.15(t,J=10.8Hz,3H). 13 CNMR(100 MHz,CDCl 3 )δ169.8,153.9,151.9,151.0,137.0,119.9,79.6,74.0,67.3,63.0,62.2,49.9,35.9,35.2,29.8,28.6,14.1.ESI-HRMS(m / z)calcd C 21 H 31 N 6 O 4 S(M+H) + ,463.2111;found,463.2120.HPLC purity:95.9%.

[0040] 1.2.18.(±)-Ethyl-4-hydroxy-3-(6-(4-methylpiperazin-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(18a,18b)

[0041] 18a:white solid,29.5%yield. 1 H NMR(600 MHz,CDCl 3)δ8.23(s,1H),8.02(s,1H),5.40(t,J=6.6 Hz,1H),4.35(s,2H),4.24-4.20(m,2H),3.74(d,J=11.4 Hz,1H),3.44(d,J=11.4 Hz,1H),3.23(dd,J=6 Hz,J=10.8 Hz,1H),2.80(dd,J=7.2Hz,J=10.8Hz,1H),2.55(t,J=5.4 Hz,4H),2.35(s,3H)1.77(s,2H),1.19(t,J=7.2 Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ168.6,154.1,151.9,150.9,137.9,119.8,73.8,63.1,55.2,46.2,35.3,33.4,14.0.ESI-HRMS(m / z)calcd C 17 H 24 ClN 6 O 3 SNa(M+Na) + ,393.1703;found,393.1701.HPLC purity:98.1%.18b:white solid,41.6%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.26(s,1H),7.93(s,1H),5.26(t,J=6 Hz,1H),4.34(s,2H),4.25-4.18(m,2H),3.88(d,J=12 Hz,1H),3.54(d,J=12 Hz,1H),3.24(q,J=5.4 Hz,1H),2.93(q,J=5.4 Hz,1H),2.56(s,4H),2.36(s,3H),1.75(s,2H),1.15(t,J=7.2 Hz,3H). 13 C NMR(150 MHz,CDCl 3 )δ169.8,154.1,151.9,151.0,137.1,119.9,79.6,74.0,63.0,55.2,46.3,35.9,35.2,14.1.ESI-HRMS(m / z)calcdC 17 H 24 ClN 6 O 3 SNa(M+Na) +,393.1703;found,393.1699.HPLCpurity:95.6%.

[0042] 1.2.22.(±)-Ethyl-3-(6-(ethylthio)-9H-purin-9-yl)-4-hydroxytetrahydrothiophene-3-carboxylate(22a,22b)

[0043] 22a:yellow solid,21.5%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.58(s,1H),8.28(s,1H),5.44(q,J=6 Hz,1H),5.12(d,J=5.4 Hz,1H),4.18-4.13(m,2H),3.73(d,J=12Hz,1H),3.65(d,J=11.4 Hz,1H),3.34-3.28(m,3H),2.86(q,J=5.4 Hz,1H),1.41(t,J=7.8Hz,3H),1.12(t,J=7.2 Hz,3H). 13 C NMR(150 MHz,CDCl 3 )δ168.2,162.3,151.5,148.4,142.3,130.9,76.5,74.0,63.0,34.8,33.9,23.4,14.7,13.9.ESI-HRMS(m / z)calcdC 14 H 18 N 4 O 3 S 2 Na(M+Na) + ,377.0713;found,377.0697.HPLC purity:99.8%.22b:yellowsolid,22.6%yield. 1 H NMR(600 MHz,CDCl 3)δ8.63(s,1H),8,28(s,1H),5.48(q,J=6 Hz,1H),4.81(d,J=5.4 Hz,1H),4.23-4.19(m,2H),3.77(d,J=12 Hz,1H),3.55(d,J=12 Hz,1H),3.38(q,J=7.2 Hz,2H),3.28(q,J=6 Hz,1H),2.85(dd,J=6.6 Hz,J=11.4 Hz,1H),1.45(t,J=7.2 Hz,3H),1.17(t,J=7.2 Hz,3H). 13 C NMR(150 MHz,CDCl 3 )δ168.2,162.9,151.5,148.5,142.2,131.3,74.0,63.2,35.1,33.6,23.5,14.8,14.0.ESI-HRMS(m / z)calcdC 14 H 18 N 4 O 3 S 2 Na(M+Na) + ,377.0713;found,377.0704.HPLC purity:99.3%.

[0044] 1.2.23.(±)-Ethyl-4-hydroxy-3-(6-(isopropylthio)-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(23a,23b)

[0045] 23a:white solid,51.2%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.63(s,1H),8.27(s,1H),5.48(s,1H),4.81(s,1H)4.37-4.33(m,1H),4.24-4.20(m,2H)3.78(d,J=12 Hz,1H),3.53(d,J=12 Hz,1H),3.28(q,J=6 Hz,1H),2.85(dd,J=6.6 Hz,J=10.8 Hz,1H),1.50(d,J=7.2 Hz,6H),1.18(t,J=7.2 Hz,3H). 13 C NMR(150 MHz,CDCl 3)δ168.3,163.2,151.5,148.5,142.1,131.2,74.0,63.3,35.1,34.7,33.6,23.4,23.3,14.0.ESI-HRMS(m / z)calcd C 15 H 20 N 4 O 3 S 2 Na(M+Na) + ,391.0869;found,391.0871.HPLC purity:99.8%.23b:whitesolid,22.4%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.61(s,1H),8.20(s,1H),5.76(s,1H),5.30(s,1H),4.34-4.29(m,1H),4.19-4.15(m,2H)3.91(d,J=12.6 Hz,1H),3.56(d,J=12.6Hz,1H),3.16(dd,J=4.8 Hz,J=11.4 Hz,1H),2.86(dd,J=5.4 Hz,J=12 Hz,1H),1.46(d,J=6.0 Hz,6H),1.09(t,J=6.6 Hz,3H). 13 C NMR(150 MHz,CDCl 3 )δ169.2,162.8,151.4,148.4,141.2,131.2,74.0,63.1,35.5,35.2,34.6,23.3,23.2,13.9.ESI-HRMS(m / z)calcd C 15 H 20 N 4 O 3 S 2 Na(M+Na) + ,391.0869;found,391.0868.HPLC purity:95.1%.

[0046] 1.2.24.(±)-Ethyl-4-hydroxy-3-(6-(prop-2-yn-1-ylthio)-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(24a,24b)

[0047] 24a:white solid,41.2%yield. 1H NMR(600 MHz,CDCl 3 )δ8.68(s,1H),8.32(s,1H),5.46(s,1H),4.65(s,1H),4.23-4.18(m,2H),4.16(d,J=2.4 Hz,2H),3.76(d,J=12Hz,1H),3.60(d,J=12 Hz,1H),3.29(q,J=6 Hz,1H),2.85(q,J=6 Hz,1H),2.22(t,J=2.4 Hz,1H),1.16(t,J=7.2 Hz,3H). 13 C NMR(150 MHz,CDCl 3 )δ168.1,160.2,151.6,148.9,142.6,131.0,79.3,74.0,71.3,63.2,35.0,33.7,17.3,14.0.ESI-HRMS(m / z)calcdC 15 H 16 N 4 O 3 S 2 Na(M+Na) + ,387.0556;found,387.0550.HPLCpurity:98.6%.24b:white solid,49.7%yield. 1 H NMR(600 MHz,CDCl 3 )δ8.71(s,1H),8.27(s,1H),5.54(s,1H),5.33(s,1H),4.23-4.19(m,4H),3.93(d,J=12.6 Hz,1H),3.57(d,J=12.6 Hz,1H),3.19(dd,J=4.8Hz,J=12 Hz,1H),2.83(dd,J=4.8Hz,J=11.4 Hz,1H),2.23(t,J=2.4 Hz,1H),1.12(t,J=7.2 Hz,3H). 13 C NMR(150MHz,CDCl 3 )δ169.4,160.3,151.6,148.7,141.7,131.4,79.3,79.1,74.1,71.3,63.3,35.6,35.4,17.3,14.0.ESI-HRMS(m / z)calcd C 15 H 16 N 4 O 3 S 2 Na(M+Na)+ ,387.0556;found,387.0566.HPLC purity:97.2%.

[0048] 1.2.31.(±)-Ethyl-3-(6-((3-chloropropyl)thio)-9H-purin-9-yl)-4-hydroxytetrahydro thiophene-3-carboxylate(31a,31b)

[0049] 31a:white solid,42.4%yield. 1 H NMR(400 MHz,CDCl 3 )δ8.59(s,1H),8.31(s,1H),5.44(s,1H),5.01(s,1H),4.20-4.13(m,2H),3.75-3.63(m,4H),3.47(t,J=6.8Hz,2H),3.29(q,J=5.6 Hz,1H),2.85(dd,J=6.4 Hz,J=11.2 Hz,1H),2.26-2.20(m,2H),1.13(t,J=7.2 Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ168.1,161.4,151.5,148.6,142.5,131.0,73.9,63.1,43.5,34.8,33.9,32.2,26.0,13.9.ESI-HRMS(m / z)calcdC 15 H 19 ClN 4 O 3 S 2 Na(M+Na) + ,425.0479;found,425.0470.HPLC purity:96.4%.31b:whitesolid,21.7%yield. 1 H NMR(400 MHz,CDCl 3)δ8.62(s,1H),8.24(s,1H),5.67(s,1H),5.31(s,1H),4.176(q,J=7.2 Hz,2H),3.92(d,J=12 Hz,1H),3.68(t,J=6.4 Hz,2H),3.56(d,J=12.4 Hz,1H),3.50(t,J=6.8 Hz,2H),3.16(dd,J=5.2 Hz,J=11.6 Hz,1H),2.86(dd,J=4.8 Hz,J=11.6 Hz,1H),2.28-2.21(m,2H),1.094(t,J=7.2 Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ169.2,161.7,151.4,148.4,141.5,131.4,79.0,74.1,63.1,43.5,35.5,35.3,32.2,26.0,13.9.ESI-HRMS(m / z)calcd C 15 H 19 ClN 4 O 3 S 2 Na(M+Na) + ,425.0479;found,425.0474.HPLC purity:95.2%.

[0050] 1.2.36.(±)-Ethyl-3-(2-fluoro-6-(propylthio)-9H-purin-9-yl)-4-hydroxytetrahydrothiophene-3-carboxylate(36a,36b)

[0051] 36a:white solid,38.5%yield. 1 H NMR(400 MHz,CDCl 3 )δ8.31,(s,1H)5.44,(t,J=6 Hz,1H),4.24(q,J=13.6 Hz,2H),3.72(d,J=11.6 Hz,1H),3.61(d,J=11.6 Hz,1H),3.34-3.29(m,2H),2.85(dd,J=11.2,J=6 Hz,1H),1.86-1.77(m,2H),1.19(t,J=6.8 Hz,3H)1.08(t,J=7.6 Hz,3H). 13 C NMR(100 MHz,CDCl 3)δ169.3,166.0(d,J C-F =16.5 Hz),159.7,158.3,152.2(d,J C-F =17.6 Hz),145.2(d,J C-F =3.2 Hz),76.4,75.7,36.0,31.9,23.9,14.1,13.6. 19 F NMR(376MHz,CDCl 3 )δ-49.78.ESI-HRMS(m / z)calcd C 15 H 19 FN 4 O 3 S 2 Na(M+Na) + ,409.0775;found,409.0775.HPLCpurity:99.4%.36b:white solid,41.5%yield. 1 HNMR(600 MHz,CDCl 3 )δ8.27(s,1H),5.29(t,J=4.2 Hz,1H),4.25-4.19(m,2H)3.92(d,J=12.6 Hz,1H)3.54(d,J=12.6 Hz,1H),3.36-3.29(m,2H),3.11(dd,J=4.2 Hz,J=12 Hz,1H),3.81(dd,J=4.8 Hz,J=12 Hz,1H),1.84-1.78(m,2H),1.14(t,J=7.2 Hz,3H),1.07(t,J=7.2 Hz,3H). 13 C NMR(100 MHz,CDCl 3 )δ169.1,166.1(Jc-F=16.5 Hz),157.6(d,Jc-F=215.0 Hz),150.1(Jc-F=17.3 Hz),141.4(Jc-F=3.2 Hz),129.8(Jc-F=4.2 Hz),78.5,73.8,63.4,35.4(Jc-F=45 Hz),31.3,22.7,14.0,13.5. 19 F NMR(376 MHz,CDCl 3 )δ-49.51.ESI-HRMS(m / z)calcd C 15 H 19 FN 4 O 3 S 2 Na(M+Na)+ ,409.0775;found,409.0769.HPLCpurity:99.4%.

[0052] 1.2.39.(±)-Ethy-3-(6-chloro-2-fluoro-9H-purin-9-yl)-4-hydroxytetrahydrothiophene-3-carboxylate(39a,39b)

[0053] 39a:brown solid,21.5%yield. 1 H NMR(600MHz,CDCl 3 )δ8.61(s,1H),5.40(t,J=6.6Hz,1H),4.28-4.21(m,2H),3.72(dd,J=12.0Hz,J=29.4Hz,2H),3.59(s,1H),3.32(q,J=6.0Hz,1H),2.82(dd,J=11.4Hz,J=6.6Hz,1H),1.18(t,J=7.2Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ167.6,145.2,76.6,73.8,63.6,53.6,34.7,33.7(dd,J=100.6),14.0. 19 F NMR(376MHz,CDCl 3 )δ-48.89.ESI-HRMS(m / z)calcd C 12 H 12 ClFN 4 O 3 SNa(M+Na) + ,369.0195;found,369.0187.HPLC purity:97.5%.39b:brown solid,32.5%yield. 1 H NMR(400MHz,CDCl 3 )δ8.51(s,1H),5.30(s,2H),4.55(s,1H),4.29-4.21(m,2H),3.97(d,J=12.8Hz,1H),3.56(d,J=12.4Hz,1H),3.12(dd,J=3.2Hz,J=11.6Hz,1H),2.78(dd,J=4.4Hz,J=11.6Hz,1H),1.14(t,J=7.2Hz,3H). 13 C NMR(100MHz,CDCl3 )δ168.7,158.0,155.8,153.8,144.2(d,J C-F =3.6),130.6,78.3,74.1,63.8,53.6,35.6,35.1,14.0. 19 F NMR(376MHz,CDCl 3 )δ-48.89.ESI-HRMS(m / z)calcd C 12 H 12 ClFN 4 O 3 SNa(M+Na) + ,369.0195;found,366.0192.HPLCpurity:99.0%.

[0054] 1.2.42.(±)-Ethy-3-(2-chloro-6-morpholino-9H-purin-9-yl)-4-hydroxytetrahydrothioph ene-3-carboxylate(42a,42b)

[0055] 42a:white solid,18.4%yield. 1 HNMR(400MHz,CDCl 3 )δ8.06(s,1H),5.42(q,J=6Hz,1H),4.33(dJ=5.2Hz,3H),4.34-4.19(m,6H),3.82(t,J=5.2Hz,4H),3.70(d,J=12.0Hz,1H),3.46(d,J=11.6Hz,1H)3.26(dd,J=6.0Hz,J=10.8Hz,1H),2.81(dd,J=6.8Hz,J=11.2Hz,1H),1.23(t,J=6.8Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ168.3,154.2,153.7,152.4,138.1,118.6,76.6,73.6,67.1,63.3,35.3,33.7,14.0.ESI-HRMS(m / z)calcdC 16 H 20 ClN 5 O 4 SNa(M+Na) +, 436.0817; found, 436.0824. HPLC purity: 97.6%. 42b: white solid, 53.2% yield. 1 H NMR (400 MHz, CDCl 3 )δ7.98(s,1H),5.49(s,1H),5.26(t,J=4.4Hz,1H),4.32-4.17(m,6H),3.88(d,J=12.4Hz,1H),3.83(t,J=4.8Hz,4H) ,3.49(d,J=12.4Hz,1H),3.17(dd,J=4.8Hz,J=11.6Hz,1H),2.85(dd,J=4.4Hz,J=11.6Hz,1H)1.18(t,J=7.2Hz,3H). 13 C NMR (100 MHz, CDCl 3 )δ169.5,154.1,153.6,152.2,137.4,118.8,79.1,73.8,67.1,63.3,35.7,35.5,14.1.ESI-HRMS(m / z)calcd C 16 H 20 C1N 5 O 4 SNa(M+Na) + ,436.0817; found,436.0817.HPLC purity:96.9%.

[0056] Embodiment 2:

[0057]

[0058]

[0059] Representative experimental operation: In a reaction bottle, 1,4-dithio-2,5-diol 18 (20 mmol, 3.0448 g), (ethoxycarbonylmethylene)triphenylphosphine (24 mmol, 8.3609 g) and dichloromethane (100 mL) were added in sequence, and the reaction was stirred at room temperature for 4 h. The reaction was completed by TLC detection, and the mixture was concentrated and purified to obtain compound 19 (colorless oil).

[0060] When there are different substituents at the 6-position of purine, the synthesis method is similar. Take 6-chloropurine as an example: under nitrogen protection, 6-chloropurine (20 mmol, 3.0814 g), triphenylphosphine (0.4 mmol, 0.0142 g), sodium acetate (4 mmol, 0.3281 g) and toluene (150 mL) were added in a round-bottom flask in sequence. After stirring evenly, glacial acetic acid (4 mmol, 0.6 mL) and ethyl propiolate (24 mmol, 2.42 mL) were added. The reaction was stirred at room temperature for 12 h. The reaction was completed by TLC detection. The mixture was extracted, dried, concentrated, and separated and purified to obtain compound 21 (white solid).

[0061] Synthesis of five-membered sulfur-containing heterocyclic nucleoside compounds 22 and 23 series: When the purine 6-position carries different substituents, the method for synthesizing five-membered sulfur-containing heterocyclic nucleoside compounds at the 9-position is similar. Take the purine 6-position as Cl and the 9-position as ethyl propiolate as an example: under nitrogen protection, compound 19 (0.2mmol, 50.4mg), sodium hydride (0.6mmol, 60.0mg) and dichloromethane (2mL) were added to the reaction bottle in sequence. After stirring evenly, compound 21 (0.3mmol, 46.9mg) was added. The reaction was stirred at room temperature for 2h. The reaction was completed by TLC detection, and the reaction was quenched with ammonium chloride aqueous solution. The mixture was extracted, dried and concentrated. After separation and purification, compounds 22 and 23 were obtained, both of which were white solids. In addition, compounds 22 and 23 are racemic. Characterization data of representative compounds are as follows:

[0062] (Rac)ethyl-3-(6-chloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothiophen e-3-carboxylate(22a,23a).white solid.73.4mg,92%yield.mp:87.8-89.2℃.22a: 1 HNMR (600MHz, CDCl 3 )δ8.71(s,1H),8.36(s,1H),4.20-4.14(m,4H),3.93-3.85(m,3H),3.15-3.11(m,1H),2.94-2.90(m ,1H),2.81(dd,J=16.8,3.6Hz,1H),2.58-2.52(m,1H),1.27(t,J=7.2Hz,3H),1.10(t,J=7.2Hz,3H); 13 C NMR (150 MHz, CDCl 3)δ171.0,167.5,151.9,151.8,151.6,142.9,132.1,75.0,63.2,61.4,45.6,37.0,34.7,34.6,14.2,13.9;ESI-HRMS(m / z)calcd C 16 H 19 ClN 4 NaO 4 S(M+Na + ),421.0708;found,421.0703.23a: 1 H NMR(600MHz,CDCl 3 )δ8.78(s,1H),8.69(s,1H),4.32-4.25(m,1H),4.24-4.17(m,1H),4.14-4.04(m,2H),3.73-3.68(m,1H),3.66-3.60(m,1H),3.50(d,J=12.6Hz,1H),3.44(dd,J=10.8,6.6Hz,1H),3.13(dd,J=16.2,2.4Hz,1H),2.47(t,J=10.8Hz,1H),1.60-1.54(m,1H),1.19(t,J=7.2Hz,3H),1.12(t,J=7.2Hz,3H); 13 C NMR(150MHz,CDCl 3 )δ171.1,167.5,152.1,151.8,144.0,131.4,73.4,63.2,61.1,46.9,40.2,34.6,33.5,29.8,14.2,13.9;ESI-HRMS(m / z)calcd C 16 H 19 ClN 4 NaO 4 S(M+Na + ),421.0708;found,421.0707.

[0063] (Rac)ethyl-3-(6-(diethylamino)-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydro-othiophene-3-carboxylate(22e,23e).Colorless oil.78.4mg,90%yield.22e: 1 HNMR(600MHz,CDCl 3)δ8.25(s,1H),7.93(s,1H),4.24-4.11(m,5H),4.06-3.84(m,4H),3.80-3.72(m,2H),3.18(dd,J=11.4,6.6Hz,1H),2.91-2.87(m,1H),2.84-2.80(m,1H),2.54-2.48(m,1H),1.30-1.23(m,9H),1.13(t,J=7.2Hz,3H); 13 C NM R(150MHz,CDCl 3 )δ171.4,168.4,153.9,152.5,150.9,135.3,120.0,73.8,62.7,61.2,45.6,37.4,34.8,34.4,14.3,14.0;ESI-HRMS(m / z)calcd C 20 H 30 N 5 O 4 S(M+H + ),436.2013;found,436.2006.23e: 1 H NMR(600MHz,CDCl 3 )δ8.36(s,1H),8.22(s,1H),4.33-4.25(m,1H),4.22-4.16(m,1H),4.12-3.70(m,6H),3.64-3.54(m,2H),3.47(d,J=12.6Hz,1H),3.38(dd,J=10.8,6.6Hz,1H),3.22(dd,J=16.8,2.4Hz,1H),2.51(t,J=10.8Hz,1H),1.63(dd,J=16.8,11.4Hz,1H),1.30(t,J=7.2Hz,6H),1.21-1.18(m,3H),1.14(t,J=7.2Hz,3H); 13 CNMR(150MHz,CDCl 3 )δ171.8,168.6,153.9,152.4,151.1,136.7,119.2,72.3,62.7,60.8,46.8,40.2,34.7,33.8,14.3,14.0;ESI-HRMS(m / z)calcd C 20 H 30 N 5 O 4 S(M+H + ),436.2013;found,436.2019.

[0064] (Rac)ethyl-4-(2-ethoxy-2-oxoethyl)-3-(6-methoxy-9H-purin-9-yl)tetrahydrothioph-ene-3-carboxylate(22f,23f).Colorless oil.66.3 mg,84%yield.22f: 1 H NMR(600MHz,CDCl 3 )δ8.48(s,1H),8.13(s,1H),4.20-4.11(m,7H),3.88-3.82(m,3H),3.14(dd,J=11.4,6.0 Hz,1H),2.89(dd,J=11.4,5.4 Hz,1H),2.80(dd,J=16.8,3.6 Hz,1H),2.54(dd,J=16.8,10.2 Hz,1H),1.25(t,J=7.2 Hz,3H),1.07(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.2,168.0,161.3,152.2,152.0,139.7,122.1,74.5,62.9,61.3,54.3,45.6,37.2,34.7,34.5,14.2,13.9;ESI-HRMS(m / z)calcd C 17 H 22 N 4 NaO 5 S(M+Na + ),417.1203;found,417.1202.23f: 1 H NMR(600 MHz,CDCl 3 )δ8.55(s,1H),8.47(s,1H),4.32-4.16(m,5H),4.11-4.03(m,2H),3.69-3.58(m,2H),3.50(d,J=12.6 Hz,1H),3.40(dd,J=11.4,6.6 Hz,1H),3.15(dd,J=16.8,3.0 Hz,1H),2.48(t,J=10.8 Hz,1H),1.57(dd,J=16.2,11.4 Hz,1H),1.19(t,J=7.2 Hz,3H),1.11(t,J=7.2 Hz,3H); 13 C NMR(150MHz,CDCl 3)δ171.4,168.1,161.4,152.4,152.0,140.8,121.3,72.9,63.0,61.0,54.4,46.8,40.1,34.6,33.6,14.3,13.9;ESI-HRMS(m / z)calcd C 17 H 22 N 4 NaO 5 S(M+Na + ),417.1203;found,417.1203.

[0065] (Rac)ethyl-4-(2-ethoxy-2-oxoethyl)-3-(6-(pyrrolidin-1-yl)-9H-purin-9-yl)tetrahyd-rothiophene-3-carboxylate(22i,23i).Colorless oil.77.2 mg,89%yield.22i: 1 H NMR(600 MHz,CDCl 3 )δ8.29(s,1H),7.93(s,1H),4.22-4.13(m,6H),3.88(d,J=12.6Hz,1H),3.82-3.70(m,4H),3.18(dd,J=11.4,6.6 Hz,1H),2.90(dd,J=11.4,6 Hz,1H),2.84-2.79(m,1H),2.53(dd,J=16.8,10.8 Hz,1H),2.10-1.95(m,4H),1.28-1.24(m,3H),1.11(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.4,168.4,153.2,152.8,150.5,135.9,120.7,73.9,62.7,61.3,45.6,37.4,34.8,34.4,14.3,14.0;ESI-HRMS(m / z)calcdC 20 H 28 N 5 O 4 S(M+H + ),434.1857;found,434.1855.23i: 1 HNMR(400 MHz,CDCl 3)δ8.35(s,1H),8.26(s,1H),4.32-4.00(m,6H),3.82-3.52(m,4H),3.48(d,J=12.8 Hz,1H),3.42-3.35(m,1H),3.20(dd,J=16.4,2.4 Hz,1H),2.53-2.45(m,1H),2.13-1.95(m,4H),1.66-1.56(m,1H),1.19(t,J=7.2 Hz,3H),1.12(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.7,168.5,153.2,152.6,150.7,137.2,119.9,72.3,62.7,60.8,46.8,40.2,34.7,33.8,14.3,14.0;ESI-HRMS(m / z)calcd C 20 H 28 N 5 O 4 S(M+H + ),434.1857;found,434.1859.

[0066] (Rac)ethyl-4-(2-ethoxy-2-oxoethyl)-3-(6-(piperidin-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3-carboxylate(22j,23j).Colorless oil.75.2 mg,84%yield.22j: 1 H NMR(400 MHz,CDCl 3 )δ8.25(s,1H),7.93(s,1H),4.26-4.12(m,8H),3.89(d,J=18 Hz,1H),3.81-3.72(m,2H),3.18(dd,J=16.8,9.0 Hz,1H),2.93-2.79(m,2H),2.52(dd,J=25.2,15.6 Hz,1H),1.78-1.65(m,6H),1.26(t,J=10.8 Hz,3H),1.11(t,J=10.8Hz,3H); 13 C NMR(150 MHz,CDCl 3)δ171.4,168.4,154.0,152.4,151.1,135.1,120.2,73.8,62.7,61.3,45.6,37.5,34.8,34.4,26.2,24.9,14.3,14.0;ESI-HRMS(m / z)calcdC 21 H 30 N 5 O 4 S(M+H + ),448.2013;found,448.2007.23j: 1 H NMR(600 MHz,CDCl 3 )δ8.36(s,1H),8.22(s,1H),4.31-4.03(m,8H),3.64-3.54(m,2H),3.47(d,J=12.6 Hz,1H),3.38(dd,J=11.4,6.6 Hz,1H),3.23-3.19(m,1H),2.49(t,J=10.8 Hz,1H),1.77-1.68(m,6H),1.62-1.58(m,1H),1.19(t,J=7.2 Hz,3H),1.12(t,J=7.1Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.7,168.5,154.0,152.3,151.3,136.4,119.4,72.3,62.7,60.8,46.8,40.2,34.7,33.8,26.3,24.9,14.3,14.0;ESI-HRMS(m / z)calcd C 21 H 30 N 5 O 4 S(M+H + ),448.2013;found,448.2015.

[0067] (Rac)ethyl-4-(2-ethoxy-2-oxoethyl)-3-(6-morpholino-9H-purin-9-yl)tetrahydrothiophene-3-caroxylate(22k,23k).Colorless oil.81.8 mg,91%yield.22k: 1 H NMR(600MHz,CDCl 3)δ8.28(s,1H),7.95(s,1H),4.34-4.12(m,7H),3.90-3.74(m,8H),3.17(dd,J=11.4,6.0 Hz,1H),2.90(dd,J=11.4,5.4 Hz,1H),2.81(dd,J=16.8,3.0 Hz,1H),2.55-2.49(m,1H),1.28-1.23(m,3H),1.11(t,J=7.2 Hz,3H); 13 C NMR(150MHz,CDCl 3 )δ171.3,168.3,154.0,152.3,151.3,135.8,120.4,74.0,67.1,62.7,61.3,45.5,37.4,34.8,34.5,14.3,14.0;ESI-HRMS(m / z)calcd C 20 H 27 N 5 NaO 5 S(M+Na + ),472.1625;found,472.1618.23k: 1 H NMR(600 MHz,CDCl 3 )δ8.39(s,1H),8.26(s,1H),4.36-4.17(m,6H),4.12-4.02(m,2H),3.85(t,J=4.8 Hz,4H),3.66-3.54(m,2H),3.47(d,J=12.6 Hz,1H),3.39(dd,J=11.4,6.6 Hz,1H),3.23-3.19(m,1H),2.48(t,J=10.8 Hz,1H),1.60(dd,J=16.2,11.4 Hz,1H),1.20(t,J=7.2 Hz,3H),1.13(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.6,168.4,154.0,152.2,151.5,137.1,119.6,72.4,67.2,62.8,60.9,46.8,40.2,34.7,33.7,14.3,13.9;ESI-HRMS(m / z)calcd C 20 H 28 N 5 O 5 S(M+H + ),450.1806;found,450.1803.

[0068] (Rac)ethyl-3-(2-chloro-6-(pyrrolidin-1-yl)-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)-tetrahydrothiophene-3-carboxylate(22m,23m).Colorlessoil.81.4 mg,87%yield.22m: 1 H NMR(600 MHz,CDCl 3 )δ7.86(s,1H),4.23-4.06(m,2H),4.16-4.07(m,4H),3.86(d,J=12.0 Hz,1H),3.72-3.66(m,4H),3.19-3.15(m,1H),2.89-2.85(m,1H),2.75(dd,J=16.8,3.0 Hz,1H),2.48(dd,J=16.8,10.2 Hz,1H),2.04(q,J=6.6Hz,2H),1.94(q,J=6.6 Hz,2H),1.24(t,J=7.2 Hz,3H),1.15(t,J=7.2 Hz,3H); 13 CNMR(150 MHz,CDCl 3 )δ171.2,168.0,153.9,153.4,151.5,136.2,119.5,73.9,62.8,61.2,49.0,47.8,45.6,37.5,34.8,34.3,26.3,24.2,14.3,14.0;ESI-HRMS(m / z)calcdC 20 H 27 ClN 5 O 4 S(M+H + ),468.1467;found,468.1461.23m: 1 H NMR(600 MHz,CDCl 3)δ8.33(s,1H),4.31-4.22(m,2H),4.18-4.04(m,4H),3.73(t,J=6.6 Hz,2H),3.60(d,J=12.6 Hz,1H),3.56-3.50(m,1H),3.43(d,J=12.6 Hz,1H),3.34(dd,J=10.8,6.6 Hz,1H),3.18(dd,J=16.2,2.4 Hz,1H),2.47-2.42(m,1H),2.10-2.04(m,2H),1.99-1.93(m,2H),1.58(dd,J=16.8,11.4 Hz,1H),1.22-1.16(m,6H); 13 CNMR(150 MHz,CDCl 3 )δ171.6,168.0,153.9,153.5,151.6,137.5,118.7,72.5,63.0,60.9,49.1,47.9,46.9,40.2,34.6,33.6,26.4,24.3,14.3,14.0;ESI-HRMS(m / z)calcdC 20 H 27 ClN 5 O 4 S(M+H + ),468.1467;found,468.1466.

[0069] (Rac)ethyl-3-(2-chloro-6-(piperidin-1-yl)-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)-tetrahydrothiophene-3-carboxylate(22n,23n).Colorless oil.85.8 mg,89%yield.22n: 1 H NMR(600 MHz,CDCl 3 )δ7.88(s,1H),4.52-4.38(m,2H),4.26-4.12(m,6H),3.90(d,J=12.0 Hz,1H),3.74-3.65(m,2H),3.19(dd,J=11.4,6.0 Hz,1H),2.89(dd,J=11.4,6.0 Hz,1H),2.81-2.76(m,1H),2.50(dd,J=16.8,10.2 Hz,1H),1.76-1.65(m,6H),1.27(t,J=7.2 Hz,3H),1.19(t,J=7.2 Hz,3H);13 C NMR(150 MHz,CDCl 3 )δ171.3,168.1,154.0,153.8,152.3,135.4,119.1,74.0,62.9,61.4,45.6,37.7,34.9,34.4,26.2,24.7,14.3,14.1;ESI-HRMS(m / z)calcd C 21 H 28 ClN 5 NaO 4 S(M+Na + ),504.1443;found,504.1449.23n: 1 H NMR(600 MHz,CDCl 3 )δ8.35(s,1H),4.35-4.05(m,7H),3.61(d,J=12.6Hz,1H),3.57-3.50(m,1H),3.43(d,J=12.6 Hz,1H),3.38-3.33(m,1H),3.23-3.18(m,1H),2.49-2.44(m,1H),1.77-1.68(m,6H),1.63-1.56(m,2H),1.24-1.18(m,6H); 13 C NMR(150MHz,CDCl 3 )δ171.6,168.0,154.0,153.7,152.4,136.7,118.2,72.5,63.1,60.9,46.9,40.3,34.7,33.7,24.7,14.3,14.0;ESI-HRMS(m / z)calcd C 21 H 28 ClN 5 NaO 4 S(M+Na + ),504.1443;found,504.1446.

[0070] (Rac)ethyl-3-(2,6-dichloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothiophene-3-carboxylate(22o,23o).Colorless oil.37.3 mg,43%yield.22o: 1 H NMR(400MHz,CDCl 3)δ8.33(s,1H),4.27-4.15(m,4H),3.88-3.81(m,3H),3.15(dd,J=11.6,6.4 Hz,1H),2.94(dd,J=11.6,5.6 Hz,1H),2.80(dd,J=16.8,3.6 Hz,1H),2.94(dd,J=16.8,9.6 Hz,1H),1.29(t,J=7.2 Hz,3H),1.84(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.0,167.3,153.3,153.0,152.4,143.6,131.3,75.4,63.5,61.6,45.9,37.3,34.9,34.7,14.3,14.1;ESI-HRMS(m / z)calcd C 16 H 18 Cl 2 N 4 NaO 4 S(M+Na + ),455.0318;found,455.0314.23o: 1 H NMR(600MHz,CDCl 3 )δ8.77(s,1H),4.3(q,J=7.2 Hz,2H),4.09(q,J=7.2 Hz,2H),3.70(d,J=13.2 Hz,1H),3.65-3.59(m,1H),3.49-3.39(m,2H),3.09(dd,J=16.2,2.4 Hz,1H),2.47(t,J=10.8 Hz,1H),1.63-1.53(m,1H),1.21(dd,J=16.2,7.2 Hz,6H); 13 C NMR(150 MHz,CDCl 3 )δ170.9,167.1,153.3,153.0,152.6,144.6,130.5,73.7,63.6,61.2,46.9,40.2,34.5,33.4,14.3,14.0;ESI-HRMS(m / z)calcdC 16 H 18 Cl 2 N 4 NaO 4 S(M+Na + ),455.0318;found,455.0323.

[0071] (Rac)methyl-3-(6-chloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothioph-ene-3-carboxylate(22p,23p).Colorless oil.57.7 mg,75%yield.22p: 1 H NMR(600MHz,CDCl 3 )δ8.72(s,1H),8.36(s,1H),4.18(q,J=7.2 Hz,2H),3.94-3.86(m,3H),3.71(s,3H),3.13(dd,J=11.4,6.0 Hz,1H),2.93(dd,J=11.4,5.4 Hz,1H),2.81(dd,J=16.8,3.6 Hz,1H),2.59-2.54(m,1H),1.30-1.26(m,3H); 13 C NMR(150MHz,CDCl 3 )δ171.1,168.2,152.0,151.8,142.8,132.3,75.1,61.5,53.8,45.8,37.1,34.9,34.7,29.8,14.3;ESI-HRMS(m / z)calcd C 15 H 17 ClN 4 NaO 4 S(M+Na + ),407.0551;found,407.0551.23p: 1 H NMR(600 MHz,CDCl 3 )δ8.80(s,1H),8.71(s,1H),4.12-4.04(m,2H),3.75(s,3H),3.71(d,J=13.2 Hz,1H),3.66-3.60(m,1H),3.50(d,J=12.6 Hz,1H),3.46(dd,J=11.4,6.6 Hz,1H),3.15-3.11(m,1H),2.50-2.45(m,1H),1.59-1.52(m,1H),1.20(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl 3)δ171.0,168.1,152.2,152.0,143.8,131.5,73.3,61.1,53.7,47.0,40.2,34.5,33.4,29.8,14.3;ESI-HRMS(m / z)calcd C 15 H 17 ClN 4 NaO 4 S(M+Na + ),407.0551;found,407.0557.

[0072] (Rac)isopropyl-3-(6-chloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothiophene-3-carboxylate(22q,23q).Colorless oil.74.3 mg,90%yield.22q: 1 H NMR(600 MHz,CDCl 3 )δ8.69(s,1H),8.35(s,1H),5.06-5.01(m,1H),4.18-4.12(m,2H),3.89-3.84(m,3H),3.13(dd,J=11.4,6.0 Hz,1H),2.92(dd,J=11.4,5.4 Hz,1H),2.80(dd,J=16.8,3.0 Hz,1H),2.54(dd,J=16.8,10.2 Hz,1H),1.26(t,J=7.2 Hz,3H),1.12(d,J=6.6 Hz,3H),1.03(d,J=6.6 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.1,166.9,152.0151.8,151.6,142.9,132.1,75.1,71.5,61.4,45.6,37.1,34.7,34.5,21.5,21.4,14.3;ESI-HRMS(m / z)calcd C 17 H 21 ClN 4 NaO 4 S(M+Na + ),435.0864;found,435.0858.23q: 1 H NMR(600 MHz,CDCl 3)δ8.77(s,1H),8.69(s,1H),5.18-5.11(m,1H),4.13-4.03(m,2H),3.70-3.60(m,2H),3.49(d,J=13.2 Hz,1H),3.43(dd,J=10.8,6.6Hz,1H),3.15(dd,J=16.2,2.4 Hz,1H),2.49-2.44(m,1H),1.61-1.55(m,1H),1.23-1.17(m,6H),1.03(d,J=6.6 Hz,3H); 13 C NMR(150 MHz,CDCl 3 )δ171.1,166.9,152.1,151.8,144.1,131.4,73.6,71.4,61.1,46.8,40.2,34.6,33.5,21.7,21.4,14.3;ESI-HRMS(m / z)calcdC 17 H 21 ClN 4 NaO 4 S(M+Na + ),435.0864;found,435.0871.

[0073] (Rac)ethyl-3-(6-chloro-2-fluoro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydr-othiophene-3-carboxylate(22r,23r).Colorless oil.35.0 mg,42%yield.22r: 1 H NMR(400 MHz,CDCl 3 )δ8.33(s,1H),4.27-4.15(m,4H),4.82-4.90(m,3H),3.14(dd,J=6.4,3.6 Hz,1H),2.94(dd,J=5.2,11.6 Hz,1H),2.80(dd,J=9.6,16.8Hz,1H),2.56(dd,J=9.6,16.8 Hz,1H),1.32-1.27(m,3H),1.21-1.16(m,3H); 13 C NMR(100MHz,CDCl 3 )δ171.0,167.3,161.3,156.9(d,J C-F=22.0 Hz),143.6,75.4,63.5,61.6,45.9,37.1,34.9,34.7,29.9,14.3,14.1; 19 F NMR(565 MHz,CDCl 3 )δ-48.32;ESI-HRMS(m / z)calcdC 16 H 18 ClFN 4 NaO 4 S(M+Na + ),439.0614;found,439.0607.23r: 1 H NMR(600 MHz,CDCl 3 )δ8.76(s,1H),4.35-4.23(m,2H),4.12-4.07(m,2H),3.71(d,J=12.6 Hz,1H),3.65-3.59(m,1H),3.50-3.41(m,2H),3.11-3.07(m,1H),2.48(t,J=10.8 Hz,1H),1.63(dd,J=11.4,16.2Hz,1H),1.17-1.23(m,6H); 13 C NMR(150MHz,CDCl 3 )δ170.9,167.0,156.9(d,J C-F =220.5Hz),153.6(d,J C-F =18 Hz),144.6,73.8,63.5,61.2,46.9,40.1,34.5,33.4,29.8,14.3,14.0; 19 F NMR(565 MHz,CDCl 3 )δ-48.52.ESI-HRMS(m / z)calcd C 16 H 18 ClFN 4 NaO 4 S(M+Na + ),439.0614;found,439.0612.

[0074] (Rac)ethyl-4-(2-ethoxy-2-oxoethyl)-3-(9H-purin-9-yl)tetrahydrothiophene-3-carb-oxylate(22s,23s).Compounds 22s(39.9 mg,0.1 mmol),Pd / C(10 wt%),and DCM(1.0 mL)were added to a reaction tube under H 2 (1 atm)atroom temperature.Whencompound 22s was completely consumed,the residue wasconcentrated and purifiedby flash column chromatography on silica gel(PE / EA=2 / 1)to give the target compound 22s as a colorless oil in 23.7 mg,65%yield.23s was prepared analogously to22s.Colorless oil.22.9 mg,63%yield.22s: 1 H NMR(400 MHz,CDCl 3 )δ9.18(s,1H),8.95(s,1H),8.34(s,1H),4.22-4.12(m,3H),3.96-3.86(m,3H),3.19-3.13(m,1H),2.93(dd,J=5.2,11.6 Hz,1H),2.86-2.79(m,1H),2.57(dd,J=10.0,16.8 Hz,1H),1.76(s,1H),1.27(t,J=7.2 Hz,3H),1.09(t,J=7.2 Hz,3H); 13 C NMR(100MHz,CDCl 3 )δ171.2,167.9,152.6,149.1,143.0,74.6,63.1,61.4,45.7,37.1,34.8,34.6,29.8,14.3,14.0,1.14;ESI-HRMS(m / z)calcd C 16 H 20 N 4 NaO 4 S(M+Na +),387.1097;found,387.1091.23s: 1 H NMR(400 MHz,CDCl 3 )δ9.07(s,1H),8.84(s,1H),8.32(s,1H),4.12-4.00(m,4H),3.90-3.76(m,3H),3.12-3.04(m,1H),2.84(dd,J=5.2,16.8 Hz,1H),2.77-2.70(m,1H),2.48(dd,J=2.8,16.8 Hz,1H),1.16(q,J=7.2Hz,3H),1.02-0.94(m,3H); 13 C NMR(100 MHz,CDCl 3 )δ171.0,167.6,151.5,148.9,142.8,134.3,74.4,62.8,61.1,45.4,36.9,34.6,34.3,14.1,13.7;ESI-HRMS(m / z)calcd C 16 H 20 N 4 NaO 4 S(M+Na + ),387.1097;found,387.1090.

[0075] (Rac)ethyl-3-(2-chloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothiophene-3-carboxylate(22t,23t).Compounds 22o(43.3 mg,0.1 mmol),Pd / C(10 wt%),andDCM(1.0 mL)were added to a reaction tube under H 2(1 atm)atroom temperature.When compound 22o was completely consumed,the residue wasconcentrated andpurified by flash column chromatography on silica gel(PE / EA=2:1)to give thetarget compound 22t as a colorless oil in 14.8 mg,37%yield.23t,Colorless oil.12.7mg,32%yield.22t: 1 H NMR(400 MHz,CDCl 3 )δ9.03(s,1H),8.37(s,1H),4.26-4.14(m,4H),3.91-3.82(m,2H),3.20-3.12(m,1H),2.97-2.90(m,1H),2.84-2.76(m,1H),2.60-2.50(m,1H),1.63(s,1H),1.28(t,J=6.8 Hz,3H)1.17(t,J=7.2Hz,3H); 13 C NMR(100 MHz,CDCl 3 )δ171.1,167.6,154.4,150.6,75.0,63.3,61.5,45.8,37.3,34.9,34.6,29.8.14.3,14.0;ESI-HRMS(m / z)calcd C 16 H 19 ClN 4 NaO 4 S(M+Na + ),421.0708;found,421.0708.23t: 1 H NMR(400 MHz,CDCl 3 )δ9.00(s,1H),8.76(s,1H),4.25-4.34(m,2H),4.09(q,J=7.2,2H),3.73-3.58(m,2H),3.52-3.38(m,2H),3.16-3.08(m,1H),2.49(q,J=10.8 Hz,1H),1.59(q,J=11.2,16.0 Hz,1H),11.23-1.15(m,6H); 13 C NMR(100MHz,CDCl 3)δ171.1,167.4,154.4,153.4,150.8,144.7,132.8,73.2,63.4,61.2,47.0,40.2,34.6,33.5,14.3,14.0;ESI-HRMS(m / z)calcdC 16 H 19 ClN 4 NaO 4 S(M+Na + ),421.0708;found,421.0702.

[0076] (Rac)ethyl-3-(2,6-dichloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothio-phene-3-carboxylate 1-oxide(22u).Compounds 22a(86.4 mg,0.2mmol),m-chloroperbenzoic acid(34.5 mg,0.2 mmol)and DCM(2 ml)were added to areaction tubeand stirred at room temperature.When compound 22a was completelyconsumed,theresidue was concentrated and purified by flash columnchromatography on silica gel(PE / EA=2 / 1)to give the target compound 22u as acolorless oil in 51.2 mg,57%yield.2u: 1 H NMR(400 MHz,CDCl 3 )δ8.46(s,1H),4.72-4.58(m,2H),4.35-4.12(m,4H),3.54-3.47(m,1H),3.22(dd,J=2.4,14.8 Hz,1H),3.02-2.90(m,2H),3.39(dd,J=10.4,16.4 Hz,1H),1.32(t,J=7.2 Hz,3H),1.16(t,J=7.2 Hz,3H); 13 CNMR(100 MHz,CDCl 3)δ177.7,169.8,167.9,152.7,142.6,64.2,63.5,62.1,42.1,34.3,14.3,14.1;ESI-HRMS(m / z)calcd C 16 H 18 Cl 2 N 4 NaO 5 S(M+Na + ),471.0267;found,471.0258.

[0077] (Rac)ethyl-3-(2,6-dichloro-9H-purin-9-yl)-4-(2-ethoxy-2-oxoethyl)tetrahydrothiophene-3-carboxylate 1,1-dioxide(22v).Compounds 22a(86.4 mg,0.2mmol),m-chloroperbenzoic acid(69.0 mg,0.4 mmol)and DCM(2 ml)were added to areactiontube and stirred at room temperature.When compound 22a was completelyconsumed,the residue was concentrated and purified by flash columnchromatography on silicagel(PE / EA=2:1)to give the target compound 22u as awhite solid in 63.3mg,68%yield.mp:118.7-120.1℃. 1 H NMR(400 MHz,CDCl 3 )δ8.36(s,1H),4.52(d,J=14.4 Hz,1H),4.40-4.05(m,5H),3.75-3.67(m,2H),3.36-3.28(m,1H),2.94-2.87(m,1H),2.44(q,J=10.4,16.4 Hz,1H),1.31-1.19(m,6H); 13 C NMR(100 MHz,CDCl 3)δ169.7,166.7,153.5,153.2,152.8,142.4,131.2,67.9,64.7,62.3,59.5,54.1,38.2,34.1,14.2,14.1; ESI-HRMS(m / z)calcd C 16 H 18 Cl 2 N 4 NaO 6 S(M+Na + ),487.0216;found,487.0206.

[0078] Example 3

[0079] 3.1 Experimental Materials

[0080] 1. Virus plasmid pZIKV-NLuc, which is a vector of plasmid pACYC177, contains the Zika virus genome sequence and the Nanoluciferase (NLuc) reporter gene. After the plasmid is transfected into Vero cells, the supernatant is harvested and infected into Vero cells to obtain the recombinant virus ZIKV-NLuc.

[0081] 2. Cells: African green monkey kidney cells (Vero), culture conditions: DMEM + 10% FBS + 1% Pen-Strep.

[0082] 3. Main reagents

[0083] DMEM (Gibco, cat:C11995500BT);

[0084] Fetal bovine serum FBS (Gibco, cat: 10270-106);

[0085] Double antibody Pen-Strep (10,000U / mL) (M&C gene biotechnology, cat: G2723M3);

[0086] Anti-Zika virus positive drug: Ribavirin (Solarbio, cat: IR0090);

[0087] Gaussia-Lumi TM Gaussia luciferase reporter gene detection kit (Beyotime, cat: RG072S)

[0088] 3.2 Research Methods (Research Methods Process Figure 1 )

[0089] 1. Preliminary screening of compounds to inhibit ZIKV-NLuc luciferase activity

[0090] Vero cells were collected at 1.5×10 4 cells / well were seeded in 96-well plates and cultured for 12 h;

[0091] When the cell confluence reaches 90-95%, discard the original culture medium, add the compound to be tested, the final concentration of the compound is 25 μM, and set up 3 replicate wells;

[0092] Incubate at 37°C for 4 h, discard the supernatant, and add ZIKV-NLuc virus dilution (MOI = 0.01);

[0093] Incubate at 37°C for 2 h, wash away free viruses, and add 100 μL of the test compound to a final concentration of 25 μM;

[0094] Incubate at 37°C for 48 h, add 100 μL of Gaussia-Lumi TM Detection reagent, detection of Nano luciferase (NLuc) by multi-function microplate reader

[0095] The experiment set up three control groups, namely, the positive drug group: adding virus and Ribavirin with a final concentration of 100 μM; the virus infection group: adding only virus without compound; the normal cell group: adding neither virus nor drug.

[0096] 3.3 Test results:

[0097] Ribavirin, a positive anti-Zika virus drug, can significantly inhibit the proliferation of Zika virus at a concentration of 100 μM. Among the series of nucleoside compounds tested, compounds 39b (LZ-39b), 22r (LZ-22r) and 22n (LZ-22n) can inhibit the replication of Zika virus on Vero cells at a concentration of 25 μM.

[0098] Three compounds with typical inhibitory effects at 25 μM concentration

[0099]

[0100]

[0101] from Figure 2 It can be seen that (±)-39b compound EC 50 =6.14μM,CC 50 =129.446μM. The selectivity coefficient of the two is SI = CC 50 / EC 50 =21.08.

[0102] from Figure 3 It can be seen that (±)-22r compound EC 50 =18.486μM,CC50 =141.864μM. The selectivity coefficient of the two is SI = CC 50 / EC 50 =7.674.

[0103] from Figure 4 It can be seen that (±)-22n compound EC 50 =28.67μM,CC 50 =316.0813μM. The selectivity coefficient of the two is SI = CC 50 / EC 50 =11.02.

[0104] Antiviral activity experiments found that compounds 39b, 22r and 22n of the present invention have good anti-Zika virus activity and have good prospects in the preparation of drugs for the treatment of Zika disease and related symptoms.

[0105] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. A five-membered sulfur heterocyclic nucleoside lead compound, including its racemate and chiral compound, characterized in that: The general structure is as follows: Where: R 1 is selected from hydrogen, halogen, amino, C1-C20 alkyl, C1-C20 alkoxy; R 2 is selected from hydrogen, halogen, C1-C20 alkyl, C1-C20 alkoxy, C1-20 alkylthio, C2-C20 alkynylthio, halogenated C1-C20 alkylthio, benzylthio, phenylthio, substituted phenylthio, di(C1-C8 alkyl)amine, X = CH2, O, S, NH, N-(C1-C4)alkyl; n = 0-5; R 3 =C1-C8 alkyl, halogen, trifluoromethyl, benzyl, substituted phenylmethyl, morpholinyl; wherein the substituent in the substituted phenyl is halogen or C1-C8 alkyl; R is selected from hydrogen, C1-C20 alkyl.

2. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 1, characterized in that: R 1 is selected from hydrogen, halogen, amino; R 2 is selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-6 alkylthio, C2-C4 alkynylthio, halogenated C1-C4 alkylthio, benzylthio, phenylthio, substituted phenylthio, di(C1-C2 alkyl)amine, X = CH2, O, S, NH, N-(C1-C4)alkyl; n = 1-3; R 3 =C1-C4 alkyl, halogen, trifluoromethyl, benzyl, substituted phenylmethyl, morpholinyl; wherein the substituent in the substituted phenyl is halogen or C1-C4 alkyl.

3. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 2, characterized in that: R is selected from ethyl.

4. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 3, characterized in that: The lead compound structure is 5. Use of the five-membered sulfur heterocyclic nucleoside lead compound and a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 in the preparation of antiviral drugs.

6. The use of the five-membered sulfur heterocyclic nucleoside lead compound and a pharmaceutically acceptable salt thereof in the preparation of an antiviral drug according to claim 5, characterized in that: Antiviral is against Zika virus.

7. The use of the five-membered sulfur heterocyclic nucleoside lead compound and a pharmaceutically acceptable salt thereof in the preparation of an antiviral drug according to claim 5, characterized in that: The pharmaceutically acceptable salts include salts formed between acyclic nucleoside compounds and organic or inorganic acids.

8. Use of the five-membered sulfur heterocyclic nucleoside lead compound and a pharmaceutically acceptable salt thereof in the preparation of an antiviral drug according to claim 7, characterized in that: The organic acid is selected from one or more of malic acid, lactic acid, camphorsulfonic acid, citric acid, fumaric acid or oxalic acid, and the organic acid is selected from one or more of phosphoric acid, hydrohalic acid, sulfuric acid or nitric acid.

9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4.

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