Five-membered sulfur-containing heterocyclic nucleoside lead compound and application thereof in resisting cervical cancer and pancreatic cancer activity

Abstract

The invention discloses a five-membered sulfur-containing heterocyclic nucleoside lead compound and application thereof in resisting cervical cancer, pancreatic cancer and other activities, and belongs to the technical field of medicinal chemistry. A series of five-membered sulfur-containing heterocyclic ring nucleoside compounds are synthesized through a two-step Michael addition reaction, and the five-membered sulfur-containing heterocyclic ring nucleoside compounds comprise a racemate, (3S, 4R) and (3R, 4S): # imgabs0 # (* is a chiral center). Results show that in in-vitro cervical cancer (HeLa), pancreatic cancer (PANC-1) and other cell tests, the anti-tumor activity of the racemate is close to that of the enantiomer configuration. Wherein the chiral five-membered sulfur-containing heterocyclic nucleoside lead compound (3R, 4S)-15ab shows relatively strong proliferation inhibition capacity in a cervical cancer cell line, and IC50 (half maximal inhibitory concentration) reaches 2.78 mu M and is obviously superior to that of control drugs 5-fluorouracil and cis-platinum (IC50 is 9.75 mu M and 7.99 mu M respectively).

Description

Technical Field

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

[0002] The research and development of anti-tumor drugs has always been one of the important directions in drug development. Synthetic 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 anti-virus 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 anti-tumor activity of five-membered sulfur heterocyclic nucleoside compounds. More and more evidence shows that viral diseases are closely related to the occurrence of tumors. Innovative research on chiral five-membered sulfur heterocyclic nucleoside anti-tumor 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 provide nucleoside lead compounds with more functional structures, the present invention discloses a five-membered sulfur heterocyclic nucleoside lead compound with anti-pancreatic cancer activity. In terms of synthesis method: a series of five-membered sulfur heterocyclic nucleoside compounds are synthesized by two-step Michael addition reaction and other reactions, and this type of structure includes three structural types: racemic, 3S, 4R-configuration and 3R, 4S-configuration.

[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 di-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, for the 3S, 4R-configuration and the 3R, 4S-configuration, chiral HPLC is used to separate the racemic compound.

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

[0014] Furthermore, in the above technical solution, the anti-cervical cancer activity is anti-HeLa activity; and 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 and the like, wherein the active ingredient comprises the 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; and 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 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 separation was used.

[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 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 thioheterocyclic nucleoside lead compounds for the design of drugs with anti-cervical cancer activity, on which further structural modifications and transformations 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] Embodiment 1:

[0029] Representative experimental operation: In a reaction bottle, 1,4-dithio-2,5-diol I (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 II (colorless oil).

[0030] When the 6-position of purine is a different substituent, the synthesis method is the same. 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 IV (white solid).

[0031]

[0032] Synthesis of five-membered sulfur-containing heterocyclic nucleoside compounds series a and series b: 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, taking the purine 6-position as Cl and the 9-position as ethyl propiolate as an example: under nitrogen protection, compound II (0.2mmol, 50.4mg), sodium hydride (0.6mmol, 60.0mg) and dichloromethane (2mL) were added to the reaction bottle in sequence, stirred evenly, and then compound IV (0.3mmol, 46.9mg) was added, stirred at room temperature for 2h, and the reaction was completed by TLC detection, quenched with ammonium chloride aqueous solution, extracted, dried and concentrated, and separated and purified to obtain compounds 1a and 1b, both of which were white solids. In addition, the a-series and b-series compounds are racemic.

[0033] The 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] Embodiment 2:

[0052] For 3S, 4R-configuration and 3R, 4S-configuration, chiral HPLC separation was used. Shimadzu LC-20ADCP-HPLC-08 was used in combination with chiralcel OZ-H (0.46cm ID×25cm L) and MeOH (100%) to separate 15aa and 15ab; flow rate 1.0mL / min, UV detection at 214nm. Enantiomers 15aa (ee>98%), 15ab (ee>98%). The peak with shorter retention time in HPLC trace is 15aa, and the longer peak is 15ab; the specific corresponding structure was confirmed by single crystal X-ray diffraction.

[0053] Embodiment 3:

[0054] The present invention adopts 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 on tumor cells. 3 cells. The test compound or positive control 5-FU was diluted to 100mmol / L with cell-grade dimethyl sulfoxide for later use. The cells in the culture flask were digested and centrifuged, and the cells were counted and spread on a 96-well plate after mixing. The test compound was diluted with culture medium and mixed thoroughly. The drug and HeLa cells were incubated together for 48h, and 110μL culture medium (containing 10μL CCK8) was added to each well for incubation. The OD value was recorded at 450nm and analyzed with 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 anti-tumor activity of the synthesized five-membered sulfur heterocyclic nucleoside compounds in vitro and in vivo, selected the best compound, and the overall structure-activity relationship. The experiment selected compound 15a with good anti-tumor activity; and synthesized two configuration compounds corresponding to 15a, 3S, 4R-configuration and 3R, 4S-configuration, and studied their anti-tumor activity by 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 none of them 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, and the IC 50 The IC values ​​of 5-fluorouracil (5-FU) and cisplatin on HeLa cells were 2.80 μM. 50 The chiral structures of the chiral five-membered sulfur heterocyclic nucleosides with 3S,4R and 3R,4S configurations were comparable to the antitumor activity of the racemic isomer 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 hepatoma cells; g PANC-1 human pancreatic cancer cells.

[0067] The above embodiments describe 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 by the above embodiments, and the above embodiments and the description only illustrate the principles of the present invention. Without departing from the scope of the principles of the present invention, the present invention may have various changes and improvements, and any structural changes of the five-membered sulfur heterocyclic nucleoside, these changes and improvements all fall within the scope of protection of the present invention.

Claims

1. A five-membered sulfur heterocyclic nucleoside lead compound, characterized in that: The general structure is as follows: Where: * represents chiral center, each general structure includes racemic, 3S, 4R-configuration and 3R, 4S-configuration; 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.

2. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 1, characterized in that: R 1 is hydrogen, halogen, C1-C4 alkoxy, benzyl, amino, C1-C4 alkylamino, or di-C1-C4 alkylamino; R 2 is hydrogen, chlorine; R 3 It is ethyl acetate, methyl acetate, and isopropyl acetate.

3. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 2, characterized in that: R 1 , R 2 All are chlorine, R 3 It is ethyl acetate.

4. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 3, characterized in that: The lead compound is of 3S, 4R-configuration or 3R, 4S-configuration.

5. The five-membered sulfur heterocyclic nucleoside lead compound according to claim 4, characterized in that: For the 3S,4R-configuration and 3R,4S-configuration, the racemic compound was separated by chiral HPLC.

6. Use of the five-membered sulfur heterocyclic nucleoside lead compound according to any one of claims 1 to 4 in the preparation of active drugs against cervical cancer and pancreatic cancer.

7. The use of the chiral five-membered sulfur heterocyclic nucleoside lead compound according to claim 6 in the preparation of active drugs for treating cervical cancer, pancreatic cancer, etc., characterized in that: The anti-cervical cancer activity is anti-HeLa activity; the anti-pancreatic cancer activity is anti-PANC-1 activity.

8. An anti-pancreatic cancer active drug, characterized in that: The active ingredients thereof include the compound according to any one of claims 1 to 4 and its corresponding pharmaceutically acceptable salt.

9. The anti-pancreatic cancer active drug according to claim 8, characterized in that: The pharmaceutically acceptable salts include salts formed between acyclic nucleoside compounds and organic or inorganic acids.

10. The anti-pancreatic cancer active drug according to claim 9, 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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