Five-membered sulfur-containing heterocyclic nucleoside lead compounds and their applications in anti-cervical cancer and pancreatic cancer activities

Abstract

The present invention discloses a five-membered sulfur-containing heterocyclic nucleoside lead compound and its application in anti-cervical cancer and pancreatic cancer activities, belonging to the field of pharmaceutical chemistry technology. A series of five-membered sulfur-containing heterocyclic nucleoside compounds were synthesized by a two-step Michael addition reaction, including racemate, (3S, 4R) and (3R, 4S): (* is a chiral center). The results showed that in in vitro cervical cancer (HeLa) and pancreatic cancer (PANC-1) cell tests, the anti-tumor activity of the racemate and the enantiomer configuration were similar. Among them: the chiral five-membered sulfur-containing heterocyclic nucleoside lead compound (3R, 4S)-15ab showed a strong ability to inhibit proliferation in cervical cancer cell lines, IC 50 reached 2.78 μM, which was significantly better than the control drugs 5-fluorouracil and cisplatin (IC 50 9.75 μM and 7.99 μM, respectively).

Description

Technical Field

[0001] The present invention relates to the screening of five-membered sulfur heterocyclic nucleoside lead compounds, in particular to the synthesis of five-membered sulfur heterocyclic nucleoside lead compounds and their application in anti-cervical cancer and pancreatic cancer activities, belonging to the technical field of medicinal chemistry. Background Art

[0002] The research and development of anti-tumor drugs has always been a key area of ​​drug development. Synthetic nucleoside compounds are structurally similar to natural nucleoside compounds and can inhibit viral or tumor cell replication after entering the human body, achieving antiviral or anti-tumor effects.

[0003] The research and development of nucleoside anti-tumor and antiviral drugs is gaining increasing attention. Five-membered sulfur heterocyclic nucleosides, a unique class of nucleoside compounds with distinctive physical and chemical properties, have become a key focus of nucleoside drug research and development.

[0004] Currently, five-membered sulfur heterocyclic nucleoside compounds are an important class of marketed antiviral drugs, but relatively little research has examined their anti-tumor activity. Growing evidence suggests a close relationship between viral diseases and tumorigenesis. Innovative research into chiral five-membered sulfur heterocyclic nucleoside anti-tumor drugs could not only break the international monopoly on nucleoside drugs but also hold significant scientific significance and potential application prospects. Summary of the Invention

[0005] To provide nucleoside lead compounds with more functional structures, this invention discloses five-membered sulfur heterocyclic nucleoside lead compounds with anti-pancreatic cancer activity. Synthetic methods: A series of five-membered sulfur heterocyclic nucleoside compounds were synthesized through reactions such as a two-step Michael addition reaction. These structures include racemic, 3S,4R-, and 3R,4S-configurations.

[0006] The chiral five-membered sulfur heterocyclic nucleoside lead compound of the present invention has the following general structural formula:

[0007] Among them, * represents the chiral center, and each general structure includes three structures: racemate, 3S,4R-configuration and 3R,4S-configuration.

[0008] Furthermore, in the above technical solution, R 1 、R 2 Each is independently selected from hydrogen, halogen, C1-C4 alkoxy, benzyl, amino, C1-C4 alkylamino, di-C1-C4 alkylamino; R 3 Select ester group.

[0009] Furthermore, in the above technical solution, R 1is hydrogen, halogen, C1-C4 alkoxy, benzyl, amino, C1-C4 alkylamino, or bis-C1-C4 alkylamino; R 2 is hydrogen, chlorine; R 3 It is ethyl acetate, methyl acetate, and isopropyl acetate.

[0010] Furthermore, in the above technical solution, R 1 、R 2 All are chlorine, R 3 It is ethyl acetate.

[0011] Furthermore, in the above technical solution, the lead compound is of 3S, 4R-configuration or 3R, 4S-configuration.

[0012] Furthermore, in the above technical solution, chiral HPLC is used to separate the racemic compound into 3S, 4R-configuration and 3R, 4S-configuration.

[0013] The second purpose of the present invention is to provide the use of the above-mentioned five-membered sulfur heterocyclic nucleoside lead compound in active drugs for treating cervical cancer, pancreatic cancer, etc.

[0014] Furthermore, in the above technical solution, the anti-cervical cancer activity is anti-HeLa activity; the anti-pancreatic cancer activity is anti-PANC-1 activity.

[0015] The present invention also provides an active drug for treating cervical cancer, pancreatic cancer, etc., wherein the active ingredient includes the above-mentioned chiral five-membered sulfur heterocyclic nucleoside lead compound and a pharmaceutically acceptable salt thereof.

[0016] 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.

[0017] 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 inorganic acid is selected from one or more of phosphoric acid, hydrohalic acid, sulfuric acid or nitric acid.

[0018] Furthermore, in the above technical solution, the anti-cervical cancer activity is anti-HeLa activity; the anti-pancreatic cancer activity is anti-PANC-1 activity.

[0019] The fourth object of the present invention is to provide a method for synthesizing the chiral five-membered sulfur heterocyclic nucleoside compound of the above-mentioned general structural formula, comprising the following steps:

[0020] A: For the racemate, the reaction equation is:

[0021]

[0022] Reaction conditions: (a) Ethyl (triphenylphosphoranylidene) acetate, DCM, 40 ° C, 4h. (b) Methyl propiolate (R 3 =CO2Me), Ethyl propiolate(R 3 =CO2Et),Propiolic acidisopropyl ester(R 3 =CO2i-Pr),PPh3,HOAC,NaOAc,Toluene,110℃,Refluex,N2.(c)II,NaH,DCM,Rt,N2.

[0023] B: For 3S,4R-configuration and 3R,4S-configuration, chiral HPLC was used for separation.

[0024] Among them, R 1 、R 2 Each is independently selected from hydrogen, halogen, C1-C4 alkoxy, benzyl, amino, C1-C4 alkylamino, di-C1-C4 alkylamino; R 3 Select ester group.

[0025] In order to study the antitumor activity of five-membered sulfur heterocyclic nucleosides, a series of five-membered sulfur heterocyclic nucleosides with different substituents at the 2 / 6 position of purine and the 1' position of the sugar ring were synthesized by two-step Michael addition reaction.

[0026] The experimental results showed that the chiral five-membered sulfur-containing heterocyclic nucleoside lead compound 15ab (3R, 4S) showed a strong ability to inhibit the proliferation of cervical cancer cell line (HeLa) in vitro. 50 The IC values ​​of the control drugs 5-fluorouracil (5-FU) and cisplatin on HeLa cells were 2.78 μM. 50 The results provide a basic structural unit for the design of chiral five-membered sulfur heterocyclic nucleoside lead compounds for the design of drugs with anti-cervical cancer activity, on which further structural modifications and improvements can be made.

[0027] Through the synthesis of chiral five-membered sulfur heterocyclic nucleosides and the evaluation of their anti-tumor activity in vitro and in vivo, we obtained anti-tumor lead compounds or preferred drug molecules with better activity, provided reasonable synthesis strategies and methods for five-membered sulfur heterocyclic nucleoside drugs, and laid a good foundation for the development of anti-tumor chiral five-membered sulfur heterocyclic nucleoside drugs with independent intellectual property rights. DETAILED DESCRIPTION

[0028] Example 1:

[0029] Representative experimental operation: In a reaction flask, 1,4-disulfide-2,5-diol I (20 mmol, 3.0448 g), (ethoxycarbonylmethylene)triphenylphosphine (24 mmol, 8.3609 g) and dichloromethane (100 mL) were added in sequence. The reaction was stirred at room temperature for 4 h. The reaction was detected by TLC. The reaction was concentrated and purified to obtain compound II (colorless oil).

[0030] The synthesis method is the same when the 6-position of purine is a different substituent. Taking 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 sequence to a round-bottom flask. 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 product was extracted, dried, concentrated, and separated and purified to obtain compound IV (white solid).

[0031]

[0032] Synthesis of Penta-membered Sulfur-Containing Heterocyclic Nucleoside Compounds Series A and Series B: When the purine 6-position carries different substituents, the synthesis of penta-membered sulfur-containing heterocyclic nucleoside compounds at the 9-position is similar. For example, using Cl at the 6-position and ethyl propiolate at the 9-position as an example: Under nitrogen, compound II (0.2 mmol, 50.4 mg), sodium hydride (0.6 mmol, 60.0 mg), and dichloromethane (2 mL) were added sequentially to a reaction flask. After stirring, compound IV (0.3 mmol, 46.9 mg) was added. The reaction was stirred at room temperature for 2 h. The reaction was completed by TLC. The reaction was quenched with ammonium chloride aqueous solution, extracted with water, dried, and concentrated. After separation and purification, compounds 1a and 1b were obtained as white solids. Both compounds in Series A and Series B were racemic.

[0033] Characterization data of representative compounds are as follows:

[0034] 92%.mp:87.8-89.2℃.1a: 1 H NMR(600MHz, CDCl3)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),2.9 4-2.90(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); 13C NMR(150MHz,CDCl3)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 ClN4NaO4S(M+Na + ),421.0708;found,421.0703.1b: 1 H NMR(600MHz,CDCl3)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,CDCl3)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 ClN4NaO4S(M+Na + ),421.0708;found,421.0707.

[0035] 78%.2a: 1 HNMR(600MHz,CDCl3)δ8.66(s,1H),8.36(s,1H),4.20-4.14(m,4H),3.92-3.85(m,3H),3.12(dd,J=11.4,6.6Hz,1H),2.94-2.90(m,1H),2.81(dd,J=16.8,3.6Hz,1H),2.56(dd,J=16.8,10.2Hz,1H),1.29-1.25(m,3H),1.11(t,J=7.2Hz,3H); 13CNMR(150MHz,CDCl3)δ171.0,167.5,151.7,150.7,143.7,142.7,134.7,75.1,63.2,61.4,45.6,37.0,34.7,34.5,14.2,13.9;ESI-HRMS(m / z)calcd C 16 H 19 BrN4NaO4S(M+Na + ),465.0203;found,465.0200.2b: 1 H NMR(600MHz,CDCl3)δ8.80(s,1H),8.65(s,1H),4.33-4.26(m,1H),4.24-4.18(m,1H),4.12-4.04(m,2H),3.70(d,J=12.6Hz,1H),3.66-3.60(m,1H),3.50(d,J=13.2Hz,1H),3.44(dd,J=11.4,6.6Hz,1H),3.13(dd,J=16.2,3.0Hz,1H),2.47(t,J=10.8Hz,1H),1.62-1.53(m,1H),1.20(t,J=7.2Hz,3H),1.13(t,J=7.2Hz,3H); 13 C NMR(150MHz,CDCl3)δ171.0,167.4,151.7,150.8,143.9,143.8,133.9,73.4,63.2,61.1,46.8,40.1,34.5,33.4,14.2,13.9;ESI-HRMS(m / z)calcd C 16 H 20 BrN4O4S(M+H + ),443.0383;found,443.0390.

[0036] 88%yield.3a: 1H NMR(600MHz,CDCl3)δ8.66(s,1H),8.16(s,1H),4.21-4.14(m,4H),3.92-3.84(m,3H),3.37(t,J=7.2Hz,2H),3.15(dd,J=11.4,6.0Hz,1H),2.92(dd,J=11.4,5.4Hz,1H),2.85(dd,J=16.8,3.0Hz,1H),2.56(dd,J=16.8,10.2Hz,1H),1.86-1.78(m,2H),1.28(t,J=7.2Hz,3H),1.13-1.06(m,6H); 13 C NMR(150 MHz,CDCl3)δ171.2,167.9,162.2,151.8,148.6,140.0,131.8,74.5,62.9,61.3,45.6,37.1,34.7,34.4,30.7,22.9,14.3,14.0,13.5;ESI-HRMS(m / z)calcdC 19 H 26 N4NaO4S2(M+Na + ),461.1288;found,461.1282.3b: 1 H NMR(600 MHz,CDCl3)δ8.62(s,1H),8.58(s,1H),4.31-4.25(m,1H),4.23-4.17(m,1H),4.11-4.04(m,2H),3.68(d,J=13.2 Hz,1H),3.70-3.58(m,1H),3.49(d,J=12.6 Hz,1H),3.43-3.32(m,3H),3.17-3.13(m,1H),2.48(t,J=10.8 Hz,1H),1.87-1.80(m,2H),1.61-1.55(m,1H),1.20(t,J=7.2 Hz,3H),1.14-1.07(m,6H); 13 C NMR(150 MHz,CDCl3)δ171.3,167.9,162.3,151.7,148.7,141.1,130.9,72.8,62.9,60.9,46.8,40.1,34.5,33.5,30.8,22.9,14.2,13.9,13.5;ESI-HRMS(m / z)calcd C 19 H 26 N4NaO4S2(M+Na + ),461.1288;found,461.1285.

[0037] 79%.mp:112.2-113.7℃.4a: 1 H NMR(600 MHz,CDCl3)δ8.27(s,1H),7.93(s,1H),4.21-4.11(m,5H),3.88(d,J=12.6 Hz,1H),3.80-3.73(m,2H),3.51(s,5H),3.17(dd,J=11.4,6.0 Hz,1H),2.91(dd,J=11.4,6.0 Hz,1H),2.82(dd,J=16.8,3.6Hz,1H),2.51(dd,J=16.8,6.6 Hz,1H),1.26(t,J=7.2 Hz,3H),1.09(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)δ171.4,168.4,155.0,152.3,150.9,135.3,120.6,73.9,62.7,61.3,45.5,37.4,34.8,34.4,14.3,14.0;ESI-HRMS(m / z)calcdC 18 H 25 N5NaO4S(M+Na + ),430.1519;found,430.1516.4b: 1 H NMR(400 MHz,CDCl3)δ8.36(s,1H),8.24(s,1H),4.30-4.14(m,2H),4.11-4.01(m,2H),3.65-3.44(m,9H),3.41-3.36(m,1H),3.23-3.17(m,1H),2.49(t,J=10.8 Hz,1H),1.64-1.55(m,1H),1.19(t,J=7.2 Hz,3H),1.11(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)δ171.6,168.5,155.0,152.1,151.0,136.6,119.7,72.3,62.6,60.8,46.7,40.1,34.6,33.7,14.2,13.9;ESI-HRMS(m / z)calcd C 18 H 26 N5O4S(M+H + ),408.1700;found,408.1700.

[0038] 90%.5a:1 H NMR(600 MHz,CDCl3)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=11.4,6.6 Hz,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=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)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)calcdC 20 H 30 N5O4S(M+H + ),436.2013;found,436.2006.5b: 1 H NMR(600 MHz,CDCl3)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=12.6 Hz,1H),3.38(dd,J=10.8,6.6 Hz,1H),3.22(dd,J=16.8,2.4 Hz,1H),2.51(t,J=10.8 Hz,1H),1.63(dd,J=16.8,11.4 Hz,1H),1.30(t,J=7.2 Hz,6H),1.21-1.18(m,3H),1.14(t,J=7.2Hz,3H); 13 C NMR(150 MHz,CDCl3)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 N5O4S(M+H + ),436.2013;found,436.2019.

[0039] 84%.6a: 1H NMR(600 MHz,CDCl3)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=7.2Hz,3H),1.07(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)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)calcdC 17 H 22 N4NaO5S(M+Na + ),417.1203;found,417.1202.6b: 1 H NMR(600 MHz,CDCl3)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=12.6Hz,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=10.8Hz,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(150 MHz,CDCl3)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 N4NaO5S(M+Na + ),417.1203;found,417.1203.

[0040] 83%yield.7a: 1H NMR(600 MHz,CDCl3)δ8.45(s,1H),8.12(s,1H),4.64(q,J=7.2Hz,2H),4.18-4.11(m,4H),3.89-3.82(m,3H),3.17-3.12(m,1H),2.91-2.87(m,1H),2.78(dd,J=16.8,3.0 Hz,1H),2.54(dd,J=16.8,10.2 Hz,1H),1.49(t,J=7.2 Hz,3H),1.27-1.21(m,3H),1.10-1.06(m,3H); 13 C NMR(150 MHz,CDCl3)δ171.2,168.0,161.0,152.2,152.1,139.5,122.0,74.5,63.3,62.9,61.3,45.6,37.1,34.7,34.4,14.6,14.2,13.9;ESI-HRMS(m / z)calcd C 18 H 24 N4NaO5S(M+Na + ),431.1360;found,431.1364.7b: 1 H NMR(600 MHz,CDCl3)δ8.54(s,1H),8.44(s,1H),4.70-4.63(m,2H),4.30-4.24(m,1H),4.23-4.16(m,1H),4.10-4.04(m,2H),3.70-3.58(m,2H),3.50(d,J=12.6 Hz,1H),3.40(dd,J=11.4,6.6 Hz,1H),3.15(dd,J=16.2,2.4Hz,1H),2.48(t,J=10.8 Hz,1H),1.61-1.49(m,4H),1.19(t,J=7.2 Hz,3H),1.11(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)δ171.4,168.1,161.1,152.4,152.0,140.6,121.3,72.9,63.4,63.0,61.0,46.8,40.1,34.6,33.6,14.6,14.3,14.0;ESI-HRMS(m / z)calcd C 18 H 24 N4NaO5S(M+Na + ),431.1360;found,431.1363.

[0041] 87%yield.8a: 1 H NMR(600 MHz,CDCl3)δ8.49(s,1H),8.14(s,1H),7.54-7.51(m,2H),7.37-7.33(m,2H),7.31-7.28(m,1H),5.65(s,2H),4.19-4.12(m,4H),3.90-3.83(m,3H),3.17-3.13(m,1H),2.90(dd,J=11.4,5.4 Hz,1H),2.79(dd,J=16.8,3.0 Hz,1H),2.55(dd,J=16.8,10.2 Hz,1H),1.25(t,J=7.2 Hz,3H),1.09(t,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)δ171.2,167.9,160.7,152.4,151.9,139.7,136.1,128.5,128.4,128.2,122.1,74.5,68.6,62.9,61.3,45.5,37.1,34.6,34.4,14.2,13.9;ESI-HRMS(m / z)calcdC 23 H 27 N4O5S(M+H + ),471.1697;found,471.1695.8b: 1 H NMR(600 MHz,CDCl3)δ8.55(s,1H),8.46(s,1H),7.56-7.53(m,2H),7.39-7.35(m,2H),7.34-7.30(m,1H),5.70(d,J=12.0 Hz,1H),5.67(dd,J=30.6,12.0 Hz,1H),4.31-4.24(m,1H),4.22-4.16(m,1H),4.10-4.04(m,2H),3.68-3.59(m,2H),3.50(d,J=13.2 Hz,1H),3.42-3.38(m,1H),3.17-3.12(m,1H),2.48(t,J=10.8 Hz,1H),1.58(dd,J=16.2,11.4 Hz,1H),1.21-1.17(m,3H),1.11(t,J=7.2Hz,3H); 13C NMR(150 MHz,CDCl3)δ171.4,168.0,160.8,152.6,151.9,140.8,136.1,128.6,128.5,128.3,121.3,72.9,68.7,62.9,60.9,46.8,40.1,34.6,33.6,14.2,13.9;ESI-HRMS(m / z)calcd C 23 H 26 N4NaO5S(M+Na + ),493.1516;found,493.1513.

[0042] 89%.9a: 1 H NMR(600 MHz,CDCl3)δ8.29(s,1H),7.93(s,1H),4.22-4.13(m,6H),3.88(d,J=12.6 Hz,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,CDCl3)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)calcd C 20 H 28 N5O4S(M+H + ),434.1857;found,434.1855.9b: 1 H NMR(400 MHz,CDCl3)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); 13C NMR(150 MHz,CDCl3)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 N5O4S(M+H + ),434.1857;found,434.1859.

[0043] 84%.10a: 1 H NMR(400 MHz,CDCl3)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=10.8 Hz,3H); 13 C NMR(150 MHz,CDCl3)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)calcd C 21 H 30 N5O4S(M+H + ),448.2013;found,448.2007.10b: 1 H NMR(600 MHz,CDCl3)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=7.1 Hz,3H); 13C NMR(150 MHz,CDCl3)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 N5O4S(M+H + ),448.2013;found,448.2015.

[0044] 91%.11a: 1 H NMR(600 MHz,CDCl3)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(150 MHz,CDCl3)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 N5NaO5S(M+Na + ),472.1625;found,472.1618.11b: 1 H NMR(600 MHz,CDCl3)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); 13CNMR(150 MHz,CDCl3)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 N5O5S(M+H + ),450.1806;found,450.1803.

[0045] 65%.12a: 1 H NMR(600 MHz,CDCl3)δ8.98(s,1H),8.78(d,J=7.2 Hz,2H),8.36(s,1H),7.60-7.51(m,3H),4.24-4.14(m,4H),3.97-3.89(m,3H),3.20(dd,J=11.4,6.0 Hz,1H),2.96(dd,J=11.4,6.0 Hz,1H),2.87(dd,J=16.8,3.0 Hz,1H),2.58(dd,J=16.8,10.2 Hz,1H),1.31-1.26(t,J=7.2 Hz,3H),1.13-1.09(m,3H); 13 CNMR(150 MHz,CDCl3)δ171.3,168.0,155.4,152.8,152.3,141.7,135.6,131.5,131.3,129.9,128.8,74.5,63.0,61.4,45.7,37.3,34.8,34.6,29.8,14.3,14.0;ESI-HRMS(m / z)calcd C 22 H 25 N4O4S(M+H + ),441.1591;found,441.1586.12b: 1H NMR(600 MHz,CDCl3)δ8.95(s,1H),8.87-8.71(m,3H),7.60-7.52(m 3H),4.33-4.22(m,2H),4.12-4.03(m,2H),3.74-3.63(m,2H),3.55(d,J=12.6 Hz,1H),3.4(dd,J=11.4,6.6 Hz,1H),3.25-3.21(m,1H),2.53(t,J=10.8 Hz,1H),1.67-1.62(m,1H),1.19(d,J=7.2 Hz,3H),1.13(d,J=7.2 Hz,3H); 13 C NMR(150 MHz,CDCl3)δ171.4,168.1,155.5,152.9,152.3,142.9,135.5,131.4,130.7,129.9,128.9,72.8,63.0,61.0,46.9,40.2,34.7,33.6,29.8,14.3,13.9;ESI-HRMS(m / z)calcdC 22 H 25 N4O4S(M+H + ),441.1591;found,441.1590.

[0046] 87%yield.13a: 1 H NMR(600 MHz,CDCl3)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=6.6 Hz,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 C NMR(150MHz,CDCl3)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)calcd C 20 H 27ClN5O4S(M+H + ),468.1467;found,468.1461.13b: 1 HNMR(600 MHz,CDCl3)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 C NMR(150 MHz,CDCl3)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)calcd C 20 H 27 ClN5O4S(M+H + ),468.1467;found,468.1466.

[0047] 89%.14a: 1 H NMR(600 MHz,CDCl3)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); 13C NMR(150 MHz,CDCl3)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)calcdC 21 H 28 ClN5NaO4S(M+Na + ),504.1443;found,504.1449.14b: 1 H NMR(600 MHz,CDCl3)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,CDCl3)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 ClN5NaO4S(M+Na + ),504.1443;found,504.1446.

[0048] 43%.15a: 1 H NMR(400MHz,CDCl3)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=16.8,3.6Hz,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); 13C NMR(150 MHz,CDCl3)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 Cl2N4NaO4S(M+Na + ),455.0318;found,455.0314.15b: 1 H NMR(600MHz,CDCl3)δ8.77(s,1H),4.3(q,J=7.2Hz,2H),4.09(q,J=7.2Hz,2H),3.70(d,J=13.2Hz,1H),3.65-3.59(m,1H),3.49-3.39(m,2H),3.09(dd,J=16.2,2.4Hz,1H),2.47(t,J=10.8Hz,1H),1.63-1.53(m,1H),1.21(dd,J=16.2,7.2Hz,6H); 13 C NMR(150MHz,CDCl3)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 Cl2N4NaO4S(M+Na + ),455.0318;found,455.0323.

[0049] 75%.16a: 1 H NMR(600MHz,CDCl3)δ8.72(s,1H),8.36(s,1H),4.18(q,J=7.2Hz,2H),3.94-3.86(m,3H),3.71(s,3H),3.13(dd,J=11.4,6.0Hz,1H),2.93(dd,J=11.4,5.4Hz,1H),2.81(dd,J=16.8,3.6Hz,1H),2.59-2.54(m,1H),1.30-1.26(m,3H); 13C NMR(150MHz,CDCl3)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 ClN4NaO4S(M+Na + ),407.0551;found,407.0551.16b: 1 H NMR(600MHz,CDCl3)δ8.80(s,1H),8.71(s,1H),4.12-4.04(m,2H),3.75(s,3H),3.71(d,J=13.2Hz,1H),3.66-3.60(m,1H),3.50(d,J=12.6Hz,1H),3.46(dd,J=11.4,6.6Hz,1H),3.15-3.11(m,1H),2.50-2.45(m,1H),1.59-1.52(m,1H),1.20(t,J=7.2Hz,3H); 13 CNMR(150MHz,CDCl3)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 ClN4NaO4S(M+Na + ),407.0551;found,407.0557.

[0050] 90%.17a: 1 HNMR(600MHz,CDCl3)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=11.4,6.0Hz,1H),2.92(dd,J=11.4,5.4Hz,1H),2.80(dd,J=16.8,3.0Hz,1H),2.54(dd,J=16.8,10.2Hz,1H),1.26(t,J=7.2Hz,3H),1.12(d,J=6.6Hz,3H),1.03(d,J=6.6Hz,3H); 13C NMR (150MHz, CDCl3) 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 ClN4NaO4S(M+Na + ),435.0864; found,435.0858.17b: 1 H NMR (600MHz, CDCl3) 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=13.2Hz,1H),3.43 (dd,J=10.8,6.6Hz,1H),3.15(dd,J=16.2,2.4Hz,1H),2.49-2.44(m,1H),1.61-1.55(m,1H),1.23-1.17(m,6H),1.03(d,J=6.6Hz,3H); 13 CNMR(150MHz, CDCl3)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 ClN4NaO4S(M+Na + ),435.0864;found,435.0871.

[0051] Example 2:

[0052] Chiral HPLC was used to separate the 3S,4R- and 3R,4S-configurations. A Shimadzu LC-20ADCP-HPLC-08 system with chiralcel OZ-H (0.46 cm ID × 25 cm L) and MeOH (100%) was used for separation. The flow rate was 1.0 mL / min, and UV detection was performed at 214 nm. Enantiomers 15aa (ee > 98%) and 15ab (ee > 98%) were identified. The shorter peak in the HPLC trace was 15aa, while the longer peak was 15ab. The structures were confirmed by single crystal X-ray diffraction.

[0053] Example 3:

[0054] The present invention uses the above method to synthesize a series of racemic, R-configuration and S-configuration five-membered sulfur heterocyclic nucleoside compounds, and uses the CCK8 method to test the cell activity of these compounds against tumor cells. 3 Cells were collected. Test compounds or the positive control 5-FU were diluted to 100 mmol / L using cell-grade dimethyl sulfoxide. Cells in the culture flask were digested and centrifuged, mixed, and counted before plating in a 96-well plate. Test compounds were diluted in culture medium and mixed thoroughly. HeLa cells were incubated with the drug for 48 hours. 110 μL of culture medium (containing 10 μL CCK8) was added to each well. OD values ​​were recorded at 450 nm and analyzed using GraphPad Pism 8 software.

[0055] The anti-tumor activity results of the five-membered sulfur heterocyclic nucleoside compounds are as follows:

[0056]

[0057]

[0058]

[0059]

[0060] a The antiproliferative activity of individual compounds against tumor cells was determined by CCK8 assay. Data are the mean of triplicate determinations. b PANC-1: human pancreatic cancer cells.

[0061] Example 4

[0062] The experiment evaluated the antitumor activity of synthesized five-membered sulfur heterocyclic nucleoside compounds in vitro and in vivo, selected the optimal compound, and analyzed the overall structure-activity relationship. Compound 15a, which showed excellent antitumor activity, was selected. Two corresponding compounds, 3S,4R- and 3R,4S-configurations of 15a, were synthesized and their antitumor activity was studied using CCK8.

[0063] Compound 15a was found to have the best anti-tumor activity against pancreatic cancer cells. b HCT-116, c HeLa, d MCF-7, e A549, f HepG-2 and gThe PANC-1 and other cancer cell lines were used to test whether compound 15a has an inhibitory effect on other cancer cells. As shown in the table below, compound 15a and its R and S configurations have certain anti-tumor activity against these cancer cells, but neither has good anti-tumor activity against cervical cancer cells. Among the human liver cancer cells (SK-hep-1), human ovarian cancer cells (SKOV-3), human cervical cancer cells (Hela), and human breast cancer cells (MCF-7), the inhibitory effect on human liver cancer cells (SK-hep-1) is better. The experimental results show that: in the racemic form, compound 15a exhibits a strong ability to inhibit the proliferation of cervical cancer cell line cells (HeLa) in vitro, IC 50 The IC values ​​of the control drugs 5-fluorouracil (5-FU) and cisplatin on HeLa cells were 2.80 μM. 50 The antitumor activities of chiral five-membered sulfur heterocyclic nucleosides with 3S,4R- and 3R,4S-configurations were comparable to those of the racemate 15a.

[0064] Inhibitory effect of compound 15a on HCT-116, HeLa, MCF-7, A549, HepG-2, and PANC-1

[0065]

[0066] a The antiproliferative activity of individual compounds against tumor cells was determined by CCK8 assay. Data are the mean of triplicate determinations. b HCT-116 human colon cancer cells; c HeLa human cervical cancer cells; d MCF-7 human breast cancer cells; e A549 human lung cancer cells; f HepG-2 human hepatocellular carcinoma cells; g PANC-1 human pancreatic cancer cells.

[0067] The above embodiments illustrate the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions merely illustrate the principles of the present invention. Various variations and improvements are possible without departing from the scope of the present invention. Any structural changes to the five-membered sulfur heterocyclic nucleoside are considered to fall within the scope of protection of the present invention.

Claims

1. A five-membered sulfur heterocyclic nucleoside lead compound, characterized in that: The compound structure is as follows: Among them: 15a includes racemate, 3S,4R-configuration and 3R,4S-configuration, 15b is only a racemate; * represents the chiral center.

2. Use of the five-membered sulfur heterocyclic nucleoside lead compound 15a as claimed in claim 1 in the preparation of an anti-cervical cancer active drug.

3. The use of the five-membered sulfur heterocyclic nucleoside lead compound according to claim 2 in the preparation of an anti-cervical cancer active drug, characterized in that: The anti-cervical cancer activity is anti-HeLa activity.

4. Use of the five-membered sulfur heterocyclic nucleoside lead compound according to claim 1 in the preparation of an anti-pancreatic cancer active drug.

5. The use of the five-membered sulfur heterocyclic nucleoside lead compound according to claim 4 in the preparation of an anti-pancreatic cancer active drug, characterized in that: The anti-pancreatic cancer activity is anti-PANC-1 activity.

6. An anti-pancreatic cancer active drug, characterized in that: The active ingredients include the five-membered sulfur heterocyclic nucleoside lead compound according to claim 1 and its corresponding pharmaceutically acceptable salt.

7. The anti-pancreatic cancer active drug according to claim 6, characterized in that: The pharmaceutically acceptable salts include salts formed by the five-membered sulfur heterocyclic nucleoside lead compound and organic or inorganic acids.

8. The anti-pancreatic cancer active 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 inorganic acid is selected from one or more of phosphoric acid, hydrohalic acid, sulfuric acid or nitric acid.

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