Aspirin-dithiocarbamate hybrid as well as preparation method and application thereof

Abstract

The invention relates to the technical field of aspirin medicinal chemistry, in particular to an aspirin-dithiocarbamate hybrid and a preparation method and application thereof.The preparation method comprises the steps that aspirin aspirin (ASP) serves as a parent body, acylating chlorination, dithiocarbamate forming and N-acylation continuous three-step reaction are carried out, in the reaction process, N-acylation reaction is carried out, and N-acylation reaction is carried out to obtain the aspirin-dithiocarbamate hybrid. A plurality of novel aspirin-dithiocarbamate hybrids (3a-l) can be efficiently synthesized only through one-time silica gel column chromatography separation and purification, and the preparation steps are simple; the toxicity of the prepared aspirin-dithiocarbamate hybrid to human lung cancer cells A549 is remarkably improved, is about 8 times of that of parent ASP, and is close to that of an anti-cancer drug irinotecan; the prepared aspirin-dithiocarbamate hybrid 3f has a certain inhibition effect on various cancer cells, and can inhibit the growth of human lung cancer A549 cells by inducing the cycle of the human lung cancer A549 cells to stop at a G0 / G1 phase.

Description

Technical Field

[0001] The invention relates to the technical field of aspirin pharmaceutical chemistry, and in particular to an aspirin-dithiocarbamate hybrid and a preparation method and application thereof. Background Art

[0002] Data released by the International Agency for Research on Cancer (IARC) under the World Health Organization showed that there will be approximately 20 million new cancer cases worldwide in 2022. Among them, lung cancer, female breast cancer and colorectal cancer accounted for the top three number of new cancer cases respectively. At present, with the deepening of cancer treatment, a large number of patients have developed drug resistance. This is because existing chemotherapy drugs or targeted drugs directly inhibit key links such as the proliferation signal pathways and DNA synthesis of cancer cells, thereby intervening in the core proliferation mechanism of cancer cells. Cancer cells are prone to develop drug resistance through gene mutations that change the target structure and activate bypass signals. Therefore, it is necessary to develop new anti-cancer drugs that are highly selective for tumor cells and do not produce drug resistance.

[0003] Aspirin is a commonly used non-steroidal anti-inflammatory drug, generally used to relieve pain, reduce fever and resist platelet aggregation. In recent years, aspirin has also been found to have certain anti-tumor activity. Its principle is to reduce the synthesis of inflammatory mediators prostaglandins by inhibiting the activity of cyclooxygenase, thereby inhibiting the proliferation and metastasis of cancer cells to a certain extent. This process does not interfere with the core proliferation mechanism of cancer cells, so it will not cause cancer cells to develop drug resistance like chemotherapy drugs or targeted drugs. However, aspirin has limited cytotoxicity to cancer cells. At this stage, it can only play an auxiliary therapeutic role in the treatment of cancer and cannot become a new anti-cancer drug. Summary of the invention

[0004] In view of this, it is necessary to provide an aspirin-dithiocarbamate hybrid and a preparation method and application thereof, by introducing active fragments with higher adaptability and better activity into aspirin to enhance the cytotoxicity of aspirin to cancer cells, so that it can become a new anticancer drug.

[0005] In a first aspect, the present invention provides an aspirin-dithiocarbamate hybrid, wherein the aspirin-dithiocarbamate hybrid has the following general structural formula:

[0006]

[0007] Wherein, R is selected from aromatic groups, hydrocarbon groups or ester groups.

[0008] Preferably, the R is selected from one of the following groups:

[0009]

[0010] In a second aspect, the present invention provides a method for preparing an aspirin-dithiocarbamate hybrid, which is used to prepare the aspirin-dithiocarbamate hybrid as described in the first aspect, comprising the following steps:

[0011] S1, aspirin and oxalyl chloride are subjected to chlorination reaction under the catalysis of a catalyst to obtain o-acetylsalicylic acid chloride 1;

[0012] S2, piperazine, carbon disulfide and halogenated hydrocarbon react chemically under the action of the first acid binding agent to obtain dithiocarbamate 2;

[0013] S3, o-acetylsalicylic acid chloride 1 and dithiocarbamate 2 are subjected to N-acylation reaction under the action of a second acid binding agent, and then separated and purified to obtain the target product aspirin-dithiocarbamate hybrid 3a-1. The synthetic route is as follows:

[0014]

[0015]

[0016] The definition of R is the same as that in the first aspect.

[0017] Preferably, the catalyst in step S1 is N,N-dimethylformamide.

[0018] Preferably, the first acid binding agent in step S2 and the second acid binding agent in step S3 are both tertiary amines.

[0019] In a third aspect, the present invention provides an application of the aspirin-dithiocarbamate hybrid as described in the first aspect, comprising:

[0020] A pharmaceutically acceptable salt of an aspirin-dithiocarbamate hybrid as described above.

[0021] A pharmaceutical composition of the aspirin-dithiocarbamate hybrid or a pharmaceutically acceptable salt thereof as described above.

[0022] The pharmaceutical preparation of aspirin-dithiocarbamate hybrid or a pharmaceutically acceptable salt thereof as described above further comprises at least one pharmaceutically acceptable excipient or carrier.

[0023] Use of the aspirin-dithiocarbamate hybrid or its pharmaceutically acceptable salt or its pharmaceutical composition or its pharmaceutical preparation as described above in the preparation of anticancer drugs.

[0024] Preferably, the cancers that can be treated by the anticancer drug include: lung cancer, liver cancer, colorectal cancer, cervical cancer, gastric cancer and primary glioblastoma.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] 1. Aspirin-dithiocarbamate hybrid (IC 50 =26.01-132.13 μM) were more cytotoxic than the parent aspirin (IC 50 =202.31 μM), among which aspirin-dithiocarbamate hybrid 3b (IC 50 =26.01 μM) and aspirin-dithiocarbamate hybrid 3c (IC 50 =26.54 μM) has the strongest cytotoxicity against human lung cancer cell A549, which is about the same as the parent aspirin (IC 50 =202.31μM), 8 times that of the anticancer drug irinotecan (IRT, IC 50 =12.36 μM), therefore, the aspirin-dithiocarbamate hybrid prepared by the present invention can effectively treat lung cancer;

[0027] 2. The preparation method of aspirin-dithiocarbamate hybrid provided by the present invention can use aspirin (ASP) as a matrix, and undergo three consecutive steps of chlorination, dithiocarbamate formation, and N-acylation. During the reaction process, only one separation and purification is performed to efficiently synthesize a number of novel aspirin-dithiocarbamate hybrids, and the preparation steps are simple;

[0028] 3. The aspirin-dithiocarbamate hybrid 3f prepared by the present invention showed good cytotoxicity to human liver cancer cells HepG2, human colorectal cancer cells HCT-116, human colorectal cancer cells SW-620, human cervical cancer cells Hela, human gastric cancer cells HGC-27 and human primary glioblastoma cells U87 (HepG2, HCT-116, SW-620, Hela, HGC-27, U87; IC 50 =48.28-142.10 μM), therefore, the aspirin-dithiocarbamate hybrid prepared by the present invention has a certain inhibitory effect on various cancer cells;

[0029] 4. Aspirin-dithiocarbamate hybrid 3f has the highest selectivity for human lung cancer A549 cells (SI LO2 / A549 =6), higher than ASP(SI LO2 / A549 =1) and IRT(SI LO2 / A549=1), and at the same time, flow cytometry detection revealed that aspirin-dithiocarbamate hybrid 3f could inhibit the growth of human lung cancer A549 cells by inducing cell cycle arrest at the G0 / G1 phase, indicating that the aspirin-dithiocarbamate hybrid 3f prepared by the present invention has great potential in the treatment of lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 is the aspirin-dithiocarbamate hybrid 3a of the present application 1 HNMR spectrum.

[0031] Figure 2 is the aspirin-dithiocarbamate hybrid 3a of the present application 13 CNMR spectrum.

[0032] Figure 3 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3a of the present application.

[0033] Figure 4 is the aspirin-dithiocarbamate hybrid 3b of the present application 1 HNMR spectrum.

[0034] Figure 5 is the aspirin-dithiocarbamate hybrid 3b of the present application 13 CNMR spectrum.

[0035] Figure 6 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3b of the present application.

[0036] Figure 7 is the aspirin-dithiocarbamate hybrid 3c of the present application 1 HNMR spectrum.

[0037] Figure 8 is the aspirin-dithiocarbamate hybrid 3c of the present application 13 CNMR spectrum.

[0038] Fig. 9 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3c of the present application.

[0039] Fig.10 is the aspirin-dithiocarbamate hybrid 3d of the present application 1 HNMR spectrum.

[0040] Fig.11 is the aspirin-dithiocarbamate hybrid 3d of the present application13 CNMR spectrum.

[0041] Fig.12 is the aspirin-dithiocarbamate hybrid 3d of the present application 19 FNMR spectrum.

[0042] Fig.13 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3d of the present application.

[0043] Fig.14 is the aspirin-dithiocarbamate hybrid 3e of the present application 1 HNMR spectrum.

[0044] Fig.15 is the aspirin-dithiocarbamate hybrid 3e of the present application 13 CNMR spectrum.

[0045] Fig.16 is the aspirin-dithiocarbamate hybrid 3e of the present application 19 FNMR spectrum.

[0046] Fig.17 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3e of the present application.

[0047] Fig.18 is the aspirin-dithiocarbamate hybrid 3f of the present application 1 HNMR spectrum.

[0048] Fig.19 is the aspirin-dithiocarbamate hybrid 3f of the present application 13 CNMR spectrum.

[0049] Fig. 20 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3f of the present application.

[0050] Fig.21 The aspirin-dithiocarbamate hybrid of the present application is 3g 1 HNMR spectrum.

[0051] Fig. 22 The aspirin-dithiocarbamate hybrid of the present application is 3g 13 CNMR spectrum.

[0052] Fig.23 The aspirin-dithiocarbamate hybrid of the present application is 3g 19 FNMR spectrum.

[0053] Fig.24 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3g of the present application.

[0054] Fig.25 is the aspirin-dithiocarbamate hybrid 3h of the present application 1 HNMR spectrum.

[0055] Fig.26 is the aspirin-dithiocarbamate hybrid 3h of the present application 13 CNMR spectrum.

[0056] Fig. 27 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3h of the present application.

[0057] Fig.28 is the aspirin-dithiocarbamate hybrid 3i of the present application 1 HNMR spectrum.

[0058] Fig.29 is the aspirin-dithiocarbamate hybrid 3i of the present application 13 CNMR spectrum.

[0059] Fig.30 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3i of the present application.

[0060] Fig.31 is the aspirin-dithiocarbamate hybrid 3j of the present application 1 HNMR spectrum.

[0061] Fig.32 is the aspirin-dithiocarbamate hybrid 3j of the present application 13 CNMR spectrum.

[0062] Fig.33 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3j of the present application.

[0063] Fig.34 is the aspirin-dithiocarbamate hybrid 3k of the present application 1 HNMR spectrum.

[0064] Fig.35 is the aspirin-dithiocarbamate hybrid 3k of the present application 13 CNMR spectrum.

[0065] Fig.36 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3k of the present application.

[0066] Fig.37 is the aspirin-dithiocarbamate hybrid 3l of the present application 1 HNMR spectrum.

[0067] Fig.38 is the aspirin-dithiocarbamate hybrid 3l of the present application 13 CNMR spectrum.

[0068] Fig.39 It is the HRMS spectrum of aspirin-dithiocarbamate hybrid 31 of the present application.

[0069] Fig.40 It is a graph showing the cell inhibition rate of the aspirin-dithiocarbamate hybrid and ASP of the present application on human lung cancer cells A549 and human normal liver cells LO2.

[0070] Fig.41 This is a graph showing the apoptotic effect of the aspirin-dithiocarbamate hybrid 3f of the present application on human lung cancer cells A549.

[0071] Fig.42 This is a diagram showing the inhibitory effect of the aspirin-dithiocarbamate hybrid 3f of the present application on human lung cancer cells A549.

[0072] Fig.43 This is a molecular docking model diagram of aspirin-dithiocarbamate hybrid 3f of the present application. DETAILED DESCRIPTION

[0073] The technical solutions and technical effects of the embodiments of the present invention are further described in detail below in conjunction with the accompanying drawings of the present invention.

[0074] In a first aspect, the present invention provides an aspirin-dithiocarbamate hybrid, and the general structural formula of the aspirin-dithiocarbamate hybrid is as follows:

[0075]

[0076] Wherein, R is selected from aromatic groups, hydrocarbon groups or ester groups.

[0077] Furthermore, R is selected from one of the following groups:

[0078]

[0079] The aspirin-dithiocarbamate hybrid (IC 50 =26.01-132.13 μM) were more cytotoxic than the parent aspirin (IC 50=202.31 μM), among which aspirin-dithiocarbamate hybrid 3b (IC 50 =26.01 μM) and aspirin-dithiocarbamate hybrid 3c (IC 50 =26.54 μM) has the strongest cytotoxicity against human lung cancer cell A549, which is about the same as the parent aspirin (IC 50 =202.31μM), 8 times that of the anticancer drug irinotecan (IRT, IC 50 =12.36 μM), therefore, the aspirin-dithiocarbamate hybrid prepared by the present invention can effectively treat lung cancer;

[0080] In a second aspect, the present invention provides a method for preparing an aspirin-dithiocarbamate hybrid, which is used to prepare the aspirin-dithiocarbamate hybrid as described in the first aspect, comprising the following steps:

[0081] S1, aspirin and oxalyl chloride are subjected to chlorination reaction under the catalysis of a catalyst to obtain o-acetylsalicylic acid chloride 1;

[0082] S2, piperazine, carbon disulfide and halogenated hydrocarbon react chemically under the action of the first acid binding agent to obtain dithiocarbamate 2;

[0083] S3, o-acetylsalicylic acid chloride 1 and dithiocarbamate 2 are subjected to N-acylation reaction under the action of a second acid binding agent, and then separated and purified to obtain the target product aspirin-dithiocarbamate hybrid 3a-1. The synthetic route is as follows:

[0084]

[0085] The definition of R is the same as that in the first aspect.

[0086] Furthermore, the catalyst in step S1 is N,N-dimethylformamide; the temperature required for the reaction in step S1 is 10° C. to 40° C.; and the time required for the reaction in step S1 is 0.5 h to 3 h.

[0087] Furthermore, the first acid binding agent in step S2 and the second acid binding agent in step S3 are both tertiary amines, for example, trimethylamine, triethylamine and tripropylamine; the temperature required for the reaction in step S2 is 10° C. to 40° C.; and the time required for the reaction in step S2 is 0.5 h to 3 h.

[0088] Furthermore, the temperature required for the reaction in step S3 is -5°C to 5°C; the time required for the reaction in step S3 is 0.5h to 2h.

[0089] The preparation method of aspirin-dithiocarbamate hybrid provided by the present invention can use aspirin (ASP) as a matrix, and carry out three consecutive steps of acyl chlorination, dithiocarbamate formation, and N-acylation. During the reaction process, only one separation and purification is performed to efficiently synthesize a plurality of novel aspirin-dithiocarbamate hybrids, and the preparation steps are simple.

[0090] In a third aspect, the present invention provides an application of the aspirin-dithiocarbamate hybrid according to the first aspect, comprising:

[0091] A pharmaceutically acceptable salt of an aspirin-dithiocarbamate hybrid as described above.

[0092] A pharmaceutical composition of the aspirin-dithiocarbamate hybrid or a pharmaceutically acceptable salt thereof as described above.

[0093] The pharmaceutical preparation of aspirin-dithiocarbamate hybrid or a pharmaceutically acceptable salt thereof as described above further comprises at least one pharmaceutically acceptable excipient or carrier; the pharmaceutical preparation comprises tablets, powders, capsules, granules or injections.

[0094] Use of the aspirin-dithiocarbamate hybrid or its pharmaceutically acceptable salt or its pharmaceutical composition or its pharmaceutical preparation as described above in the preparation of anticancer drugs.

[0095] Furthermore, cancers that can be treated by anticancer drugs include: lung cancer, liver cancer, colorectal cancer, cervical cancer, gastric cancer and primary glioblastoma.

[0096] Example 1, the general synthesis method of aspirin-dithiocarbamate hybrids is as follows:

[0097] Experimental Materials: Commercial reagents were purchased from Adamas and Anergy and used as received unless otherwise stated. Dichloromethane was redistilled dichloromethane (DCM). Thin layer chromatography (TLC) was performed on silica GF254 plates. IR spectra were recorded on a FTIR-8400S spectrometer in the form of KBr pellets. The wavelengths were obtained using a Bruker Avance III 400 MHz spectrometer. 1 HNMR and 13 C NMR spectra. Chemical shifts (δ) are in ppm with respect to tetramethylsilane. The residual solvent signal was used as 1 HNMR and 13 CNMR spectroscopy reference (CDCl3:δ H =7.26ppm,δ C=77.16ppm). The high resolution mass spectrometry (HRMS) of the target compound was tested on a Thermo Fisher LTQ Orbitrap XL. The biological activity test was conducted by the Green Pharmaceutical Collaborative Innovation Center of Zhejiang University of Technology.

[0098] Synthesis process: S101, at 25°C, aspirin (0.6 g, 3.0 mmol), DCM (10 mL), N,N-dimethylformamide (0.6 mmol / six drops), and oxalyl chloride (1.3 mL, 15.0 mmol) were added to a 50 mL flask in sequence, and the mixture was stirred for 1.5 h and concentrated under reduced pressure to obtain a crude product 1;

[0099] S201, at 25°C, piperazine (0.4 g, 4.5 mmol), DCM (10 mL), triethylamine (0.9 mL, 6.0 mmol) were added to a 50 mL flask in sequence, and a solution of CS2 (0.2 mL, 3.3 mmol) dissolved in DCM (10 mL) was slowly dripped therein, stirred for 1 h, halogenated hydrocarbon RX (3.3 mmol) was added, and the reaction was detected by TLC (reaction time was about 1.5 h), 50 mL of water was added to the reaction solution, washed with saturated brine (30 mL x 3), extracted with DCM (30 mL x 3), the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to remove excess solvent to obtain a crude product 2;

[0100] S301, at 0°C, take a 50mL flask and add crude product 2, DCM (10mL) and triethylamine (0.9mL, 6.0mmol) in sequence, then slowly drop crude product 1 in DCM (10mL) solution. Stir at room temperature, and monitor the reaction by TLC (stirring time is about 1h). Add 30mL of water to the reaction solution, wash with saturated brine (30mLⅹ3), extract with DCM (30mLⅹ3), dry the organic phase with anhydrous Na2SO4, filter, concentrate to remove excess solvent, and separate and purify the residue by silica gel column chromatography to obtain the target product aspirin-dithiocarbamate hybrid 3;

[0101] The eluent used in the silica gel column chromatography is a mixture of methanol and dichloromethane in a ratio of 1:80. The R group in the halogenated hydrocarbon RX is

[0102] The target products 3 obtained from the halogenated hydrocarbon RX of the above structural formula are numbered 3a, 3b, 3c, 3d, 3e, 3f, 3g, 3h, 3i, 3j, 3k and 3l in sequence; the target product 3 obtained by the above synthesis method is aspirin-dithiocarbamate hybrid (3a-l).

[0103] Please refer to Figures 1 to 39 ,pass1 H NMR, 13 C NMR, 19 The product prepared in Example 1 was characterized by means of F NMR, IR and HRMS.

[0104] Compound 3a: 2-acetylphenyl 4-((benzylthio)carbonothioyl)piperazine-1-carboxylate(3a). Yellow solid, yield 35% for three steps, mp192.7-193.8℃; 1 HNMR(400MHz, CDCl3)δ7.46-7.41(m,1H),7.38(d,J=7.3Hz,2H),7.33-7.26(m,5H),7.1 5(d,J=8.2Hz,1H),4.57(s,2H),4.34-3.74(m,6H),3.46(t,J=5.4Hz,2H),2.26(s,3H); 13 CNMR(101MHz,CDCl3)δ197.13,169.05,167.03,147.37,135.73,130.95,129.41,1 28.67,128.52,127.69,126.09,123.10,46.31,42.14,41.25,21.06; IR(KBr)ν / cm -1 :2925,1766,1633,1417,1193,995,746,700; HRMS(ESI)calcd for C 21 H 23 N2O3S2[M+H] + 415.1145, found 415.1146.

[0105] Compound 3b: 2-acetylphenyl 4-(((4-methylbenzyl)thio)carbonothioyl)piperazine-1-carboxylate(3b). Yellow solid, yield 34% for three steps, mp187.6-187.9℃; 11H NMR (400 MHz, CDCl3) δ 7.45 (dt, J = 8.7, 4.6 Hz, 1H), 7.31 - 7.27 (m, 3H), 7.14 (m, J = 14.4, 7.9 Hz, 4H), 4.53 (s, 2H), 3.85 (s, 6H), 3.48 (t, J = 5.3 Hz, 2H), 2.33 (s, 3H), 2.27 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 197.54, 169.11, 167.17, 147.42, 137.49, 132.36, 131.00, 129.31, 128.46, 127.64, 126.06, 123.11, 46.44, 42.09, 41.30, 21.19, 21.03; IR (KBr) ν / cm -1 : 2917, 1766, 1637, 1604, 1415, 1280, 1189, 993, 750; HRMS (ESI) calcd for C 23 H 25 N2O3S2 [M + H] + 429.1301, found 429.1295.

[0106] Compound 3c: 2-acetylphenyl 4-(((4-cyanobenzyl)thio)carbonothioyl)piperazine-1-carboxylate (3c). Light yellow solid, yield 34% for three steps, m.p. 176.7 - 179.6 °C; 1 1H NMR (400 MHz, CDCl3) δ 7.59 (d, J = 8.0 Hz, 2H), 7.49 (d, J = 8.0 Hz, 2H), 7.44 (dt, J = 8.0, 3.8 Hz, 1H), 7.28 (d, J = 4.6 Hz, 2H), 7.16 (d, J = 8.1 Hz, 1H), 4.64 (s, 2H), 4.31 - 3.83 (m, 6H), 3.51 (d, J = 13.1 Hz, 2H), 2.27 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.16, 169.13, 167.28, 147.57, 142.27, 132.40, 131.18, 130.14, 128.47, 127.69, 126.15, 123.23, 118.80, 111.42, 46.52, 41.34, 41.03, 21.14; IR (KBr) ν / cm -1: 2917, 2227, 1160, 1637, 1411, 1193, 993, 748; HRMS(ESI) calcd for C 22 H 22 N3O3S2 [M + H] + 440.1097, found 440.1096.

[0107] Compound 3d: 2-acetylphenyl 4-(((4-(trifluoromethyl)benzyl)thio)carbonothioyl)piperazine-1-carboxylate (3d). Yellow solid, yield 34%, for three steps, m.p. 198.7 - 199.8 °C; 1 1H NMR (400 MHz, CDCl3) δ 7.56 (d, J = 7.9 Hz, 2H), 7.50 (d, J = 8.1 Hz, 2H), 7.47 - 7.42 (m, 1H), 7.30 - 7.26 (m, 2H), 7.16 (d, J = 8.2 Hz, 1H), 4.64 (s, 2H), 3.85 (s, 6H), 3.49 (s, 2H), 2.26 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.38, 169.09, 167.10, 147.42, 140.56, 130.99, 129.67, 129.42, 128.45, 127.63, 126.05, 125.46, 123.09, 122.75, 46.34, 41.22, 40.99, 20.94; 19 19F NMR (376 MHz, CDCl3) δ -106.12; IR(KBr) ν / cm -1 : 2925, 1766, 1641, 1457, 1342, 1389, 1187, 1114, 1064, 993, 752; HRMS(ESI) calcd for C 22 H 22 F3N2O3S2 [M + H] + 483.1018, found 483.1013.

[0108] Compound 3e: 2-acetylphenyl 4-(((3,5-bis(trifluoromethyl)benzyl)thio)carbonothioyl)piperazine-1-carboxylate (3e). Yellow solid, yield 37% for three steps, m.p. 214.5 - 215.6 °C; 1 H NMR (400 MHz, CDCl3) δ 7.87 (s, 2H), 7.78 (s, 1H), 7.45 (s, 1H), 7.30 (s, 2H), 7.16 (s, 1H), 4.72 (t, J = 4.8 Hz, 2H), 4.10 (d, J = 164.5 Hz, 6H), 3.52 (s, 2H), 2.27 (t, J = 4.6 Hz, 3H); 13 C NMR (101 MHz, CDCl3) δ 195.54, 169.20, 167.20, 147.51, 139.56, 131.54, 129.57, 128.44, 127.66, 126.09, 124.60, 123.15, 121.89, 121.48, 46.44, 41.25, 40.25, 21.00; 19 F NMR (376 MHz, CDCl3) δ -105.93; IR (KBr) ν / cm -1 : 2917, 1764, 1643, 1421, 1371, 1276, 1122, 997, 752; HRMS (ESI) calcd for C 23 H 21 F6N2O3S2 [M + H] + 551.0892, found 551.0890.

[0109] Compound 3f: 2-acetylphenyl 4-(((4-nitrobenzyl)thio)carbonothioyl)piperazine-1-carboxylate (3f), Yellow solid, yield 34% for three steps, m.p. 157.2 - 158.9 °C; 1HNMR(400MHz,CDCl3)δ8.15(d,J=8.3Hz,2H),7.55(d,J=8.3Hz,2H),7.45(dt,J=8.6,4.5Hz,1H),7.28(d,J=4.6Hz,2H),7.15(d,J=8.2Hz,1H),4.69(s,2H),4.44-3.72(m,6H),3.50(d,J=5.7Hz,2H),2.26(s,3H); 13 C NMR(101MHz,CDCl3)δ195.67,168.90,166.92,147.20,146.93,144.36,130.86,129.99,128.25,127.46,126.16,123.47,122.95,46.15,45.55,40.27,20.78;IR(KBr)ν / cm -1 :3000,1764,1637,1517,1413,1342,1191,993,748;HRMS(ESI)calcd for C 21 H 22 N3O5S2[M+H] + 460.0995,found 460.0992.

[0110] Compound 3g: 2-acetylphenyl 4-(((4-fluorobenzyl)thio)carbonothioyl)piperazine-1-carboxylate(3g). Yellow solid, yield 36% for three steps, m.p. 198.7 - 199.3℃; 1 H NMR(400MHz,CDCl3)δ7.46 - 7.40(m,1H),7.34(dd,J = 8.3,5.4Hz,2H),7.28(d,J = 4.5Hz,2H),7.15(d,J = 8.2Hz,1H),6.98(t,J = 8.5Hz,2H),4.54(s,2H),4.46 - 3.64(m,6H),3.46(t,J = 5.1Hz,2H),2.25(s,3H); 13 C NMR(101MHz,CDCl3)δ196.73,169.63,168.97,160.76,147.22,132.12,131.52,130.86,128.20,127.51,125.90,122.93,115.21,46.21,41.00,20.78; 1919F NMR (376 MHz, CDCl3) δ -103.37; IR (KBr) ν / cm -1 : 2929, 1762, 1639, 1508, 1411, 1199, 1191, 1157, 991, 748; HRMS (ESI) calcd for C 21 H 22 FN2O3S2 [M + H] + 433.1051, found 433.1046.

[0111] Compound 3h: 2-acetylphenyl 4-((((6-cyanopyridin-3-yl)methyl)thio)carbonothioyl)piperazine-1-carboxylate (3h). Yellow solid, yield 36% for three steps, m.p. 198.3 - 198.7 °C; 1 1H NMR (400 MHz, CDCl3) δ 8.73 (s, 1H), 7.89 (d, J = 6.7 Hz, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.46 (dt, J = 4.8 Hz, 1H), 7.28 (t, J = 4.1 Hz, 2H), 7.16 (d, J = 8.2 Hz, 1H), 4.67 (s, 2H), 3.93 (d, J = 59.8 Hz, 6H), 3.51 (s, 2H), 2.27 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 195.16, 168.12, 167.28, 151.92, 147.57, 137.71, 132.55, 131.20, 130.28, 128.16, 127.67, 126.14, 123.21, 117.25, 41.28, 38.91, 37.76, 21.14; IR (KBr) ν / cm -1 : 2919, 1760, 1637, 1463, 1189, 991, 746; HRMS (ESI) calcd for C 21 H 21 N4O3S2 [M + H] + 441.1050, found 441.1049.

[0112] Compound 3i: 2-acetylphenyl 4-((allylthio)carbonothioyl)piperazine-1-carboxylate (3i). Yellow solid, yield 38% for three steps, m.p. 210.9 - 211.3 °C; 1 1H NMR (400 MHz, CDCl3) δ 7.45 (dt, J = 8.8, 4.6 Hz, 1H), 7.28 (d, J = 4.6 Hz, 2H), 7.16 (d, J = 8.2 Hz, 1H), 5.96 - 5.87 (m, 1H), 5.31 (d, J = 17.0 Hz, 1H), 5.17 (d, J = 10.0 Hz, 1H), 3.93 (dd, J = 64.5, 6.5 Hz, 8H), 3.48 (t, J = 5.4 Hz, 2H), 2.26 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 197.00, 168.96, 167.01, 147.27, 132.16, 130.87, 128.37, 127.55, 125.97, 122.99, 118.88, 46.28, 41.16, 40.20, 20.89; IR (KBr) ν / cm -1 : 2919, 2360, 1760, 1227, 1415, 1274, 1187, 997, 748; HRMS (ESI) calcd for C 17 18 21 H2N2O3S2 [M + H]+ + 365.0988, found 365.0987.

[0113] Compound 3j: 2-acetylphenyl 4-(((2-methoxy-2-oxoethyl)thio)carbonothioyl)piperazine-1-carboxylate (3j). Light yellow solid, yield 39% for three steps, m.p. 105.1 - 106.7 °C; 1 1H NMR (400 MHz, CDCl3) δ 7.44 (dt, J = 8.7, 4.5 Hz, 1H), 7.28 (d, J = 4.5 Hz, 2H), 7.15 (d, J = 8.2 Hz, 1H), 4.17 (s, 8H), 3.75 (s, 3H), 3.49 (t, J = 5.4 Hz, 2H), 2.26 (s, 3H); 1313C NMR (101 MHz, CDCl3) δ 195.52, 168.88, 168.65, 166.92, 147.14, 130.83, 128.19, 127.47, 125.93, 120.57, 51.82, 46.07, 45.79, 38.55, 20.84; IR (KBr) ν / cm -1 : 3004, 2354, 1751, 1635, 1423, 1276, 997, 750; HRMS (ESI) calcd for C 17 H 21 N2O5S2 [M+H] + 397.0886, found 397.0882.

[0114] Compound 3k: 2-acetylphenyl 4-(((2-ethoxy-2-oxoethyl)thio)carbonothioyl)piperazine-1-carboxylate (3k). Light yellow solid, yield 39% for three steps, m.p. 109.3 - 109.5 °C; 1 1H NMR (400 MHz, CDCl3) δ 7.49 - 7.42 (m, 1H), 7.29 (d, J = 7.2 Hz, 2H), 7.16 (d, J = 8.1 Hz, 1H), 4.34 - 3.86 (m, 10H), 3.49 (t, J = 5.3 Hz, 2H), 2.26 (s, 3H), 1.29 (t, J = 7.2 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 194.80, 168.29, 167.47, 146.58, 130.15, 127.82, 127.10, 125.34, 122.30, 61.05, 45.39, 40.42, 38.26, 20.20, 13.52; IR (KBr) ν / cm -1 : 2987, 1733, 1629, 1461, 1419, 1145, 995; HRMS (ESI) calcd for C 18 H 23 N2O5S2 [M+H] + 411.1043, found 411.1039.

[0115] Compound 3l: 2-acetylphenyl 4-(((2-(benzyloxy)-2-oxoethyl)thio)carbonothioyl)piperazine-1-carboxylate (3l). Yellow solid, yield 39% for three steps, mp191.7-192.4℃; 1 HNMR (400MHz, CDCl3) δ7.45-7.28(m,8H),7.15(t,J=9.9Hz,1H),5.17(s,2H),4.30-3.76(m,9H),3.54-3.37(m,1H),2.24(s,3H); 13 C NMR (101MHz, CDCl3) δ195.56,169.01,168.18,167.01,147.32,135.31,130.93,128. 50,128.32,127.57,125.98,123.02,67.52,46.22,41.09,38.90,20.92;IR(KBr)ν / cm -1 :2931,1737,1635,1341,1280,1187,1149,995; HRMS(ESI)calcd for C 23 H 25 N2O5S2[M+H] + 473.1199,found,473.1198.

[0116] Example 2, Cytotoxicity Experiment of Aspirin-Dithiocarbamate Hybrid 3

[0117] Experimental materials: Irinotecan (IRT), an anticancer drug with a similar structure to aspirin-dithiocarbamate hybrid 3, was selected as a positive control. The cytotoxicity of aspirin-dithiocarbamate hybrids 3a-l on human lung cancer cells A549 and human normal liver cells LO2 was tested by MTT assay.

[0118] Experimental method: Human lung cancer cells A549 or human normal liver cells LO2 (5×10 3, 100 μL) were seeded in a 96-well plate in triplicate, and after culturing at 37°C for 24 h, the suspension was replaced with fresh medium containing different doses of aspirin-dithiocarbamate hybrids 3a-l, ASP, and IRT (0.01 μM, 0.1 μM, 1 μM, 10 μM, and 100 μM), and the same volume of dimethyl sulfoxide (DMSO) was added to the drug-free medium negative control wells and solvent control wells; then 10 μL of DMSO was added to each well. 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) (5 mg / mL, phosphate buffered saline (PBS)) was added, and the culture medium was removed after 4 h of incubation. The MTT formazan precipitate was then dissolved in 100 μL DMSO and mechanically shaken for 10 min. The plate reader (FlexStation3, Molecular Devices) was used to read the data at a wavelength of 570 nm. At the same time, the IC values ​​of the hybrids 3a-l were calculated using the graphpad software. 50 The results are shown in Table 1 below:

[0119] Table 1

[0120]

[0121]

[0122] Analysis of the data in the table shows that aspirin-dithiocarbamate hybrids 3a-l (IC 50 =26.01-55.75 μM) had a significantly higher cytotoxicity against human lung cancer cell A549 than the parent ASP (IC 50 =202.31 μM), aspirin-dithiocarbamate hybrid 3k (IC 50 =132.13 μM). When R is 4-methylbenzyl (3b, IC 50 =26.01 μM) and 4-cyanobenzyl (3c, IC 50 =26.54μM) was the most cytotoxic, which was about the parent ASP (IC 50 =202.31 μM), 8 times higher than the positive control IRT (IC 50 =12.36 μM). Aspirin-dithiocarbamate hybrid 3f (SI=6) has the highest selectivity for human lung cancer cells (LO2 / A549), which is higher than ASP (SI=1) and IRT (SI=1).

[0123] Calculate the cell inhibition rate and plot it as Fig.40 The line graph and bar graph shown, wherein cell inhibition rate = [A570 (negative control well) - A570 (drug administration well)] / A570 (negative control well) x 100%.

[0124] from Fig.40 The data in the figure show that as the concentration of aspirin-dithiocarbamate hybrids 3a-l gradually increases, the inhibition rate of human lung cancer cell A549 gradually increases. When the concentration is 100 μM, the inhibition rates of aspirin-dithiocarbamate hybrids 3b, 3c and 3f on human lung cancer cell A549 are as high as 92.06%, 78.32% and 79.92%, respectively, while the inhibition rate of the parent ASP on human lung cancer cell A549 is only 49.38%. It can be seen that the introduction of dithiocarbamate into aspirin can significantly enhance the cytotoxicity of the parent aspirin on human lung cancer cell A549.

[0125] Example 3, Cytotoxicity Experiment of Aspirin-Dithiocarbamate Hybrid 3f on Different Cancer Cells

[0126] The in vitro cytotoxicity of aspirin-dithiocarbamate hybrid 3f against six other human cancer cells, including human liver cancer cells HepG2, human colorectal cancer cells HCT-116, human colorectal cancer cells SW-620, human cervical cancer cells Hela, human gastric cancer cells HGC-27 and human primary glioblastoma cells U87, was studied. The results are shown in Table 2 below:

[0127] Table 2

[0128]

[0129] Analysis of the data in Table 2 shows that aspirin-dithiocarbamate hybrid 3f has a significant inhibitory effect on the above six human cancer cells (IC 50 =48.28-142.10μM) have good cytotoxicity and have certain therapeutic effects on various cancer cells; at the same time, aspirin-dithiocarbamate hybrid 3f has a certain therapeutic effect on human lung cancer cells A549 (IC 50 =31.08 μM) was higher than the above six human cancer cells, demonstrating that aspirin-dithiocarbamate hybrid 3f has greater potential in the treatment of lung cancer.

[0130] Example 4: Experiment on aspirin-dithiocarbamate hybrid 3f inducing apoptosis in human lung cancer A549 cells

[0131] Experimental method: Human lung cancer A549 cells (5×10 4, 1000 μL) were inoculated in triplicate in 12-well plates, and after culturing at 37°C for 24 h, the old culture medium was replaced with fresh culture medium containing different concentrations of aspirin-dithiocarbamate hybrid 3f (20, 80 and 160 μM), and at the same time, the same volume of dimethyl sulfoxide was added to the drug-free medium negative control well and the solvent control well; then cultured at 37°C for 48 h, and after the culture medium was dried, 100 μL of preheated 0.25% trypsin without ethylenediaminetetraacetic acid (EDTA) was added to each well; after trypsinization, the treated cells were stained using the AnnexinV-FITC / PI cell apoptosis detection kit according to the manufacturer's instructions; then incubated in the dark at room temperature for 5-15 minutes, and apoptotic cells were analyzed by V-FITC / PI flow cytometer, and the results are shown in Figure 2. Fig.41 shown.

[0132] right Fig.41 Analysis of Figure A shows that with the increase in the concentration of aspirin-dithiocarbamate hybrid 3f, the number of early and late apoptotic cells gradually increased, while the number of normal cells gradually decreased; Fig.41 Figure B shows that when the concentration of aspirin-dithiocarbamate hybrid 3f is 20, 80 and 160 μM, the total apoptosis rate of human lung cancer A549 cells is 35.54%, 39.58% and 60.70%, respectively, while the control group is only 6.61%. The above results show that aspirin-dithiocarbamate hybrid 3f induces apoptosis of human lung cancer A549 cells in a concentration-dependent manner.

[0133] Example 5, Experiment on the inhibition of growth of human lung cancer A549 cells by aspirin-dithiocarbamate hybrid 3f

[0134] Experimental method: Human lung cancer A549 cells (2.5×10 5 , 2500 μL) were inoculated in triplicate in 6-well plates, and after culturing at 37°C for 24 h, the old medium was replaced with fresh medium containing different concentrations of aspirin-dithiocarbamate hybrid 3f (5, 10 and 20 μM), and at the same time, the same volume of dimethyl sulfoxide was added to the drug-free medium negative control well and the solvent control well; then the cells were cultured at 37°C for 24 h, and after the medium was dried, 200 μL of 0.25% trypsin was added to each well; after trypsin treatment, the cells were fixed with 70% ethanol at 4°C overnight, and then stained using the PI cell cycle analysis kit according to the manufacturer's instructions; finally, the cells were cultured in the dark at 37°C for 30 min, and the cell cycle was immediately analyzed using a V-FITC / PI flow cytometer, and the results were as shown in FIG. Fig.42 shown.

[0135] right Fig.42The analysis showed that as the concentration of aspirin-dithiocarbamate hybrid 3f gradually increased, the number of human lung cancer A549 cells with cell cycle arrest gradually increased. When the concentration of aspirin-dithiocarbamate hybrid 3f was 5, 10 and 20 μM, human lung cancer A549 cells gradually arrested in the G0 / G1 phase. The above results indicate that aspirin-dithiocarbamate hybrid 3f can induce human lung cancer A549 cell cycle arrest in the G0 / G1 phase, thereby inhibiting the growth of human lung cancer A549 cells.

[0136] Example 6, Analysis of Binding Properties of Aspirin-Dithiocarbamate Hybrid 3f to Receptor HER2

[0137] Experimental materials: The molecules were drawn using ChemDraw 20.0 software and minimized using MOE (Molecular Operating Environment) software. The receptor HER2 (PDBID: 3PP0) was downloaded from the Protein Data Bank (https: / / www.rcsb.org) and prepared using MOE software. The pocket in HER2 was set in the solvent environment.

[0138] Experimental methods: The molecules were docked using the Molecular Operating Environment (MOE). After setting the method (placement: triangle matcher, refinement: rigid receptor), score (placement: LondondG, refinement: GBVI / WSAdG) and pose (placement: 300, refinement: 5), the docking was performed by MOE software. The docking conformation with the lowest binding energy was selected from the 300 conformations as the representative binding model of the corresponding compound. Fig.43 shown.

[0139] right Fig.43 The analysis showed that aspirin-dithiocarbamate hybrid 3f was well wrapped by the active pocket of HER2 receptor. Furthermore, the O atoms of the carbonyl and ester groups in aspirin-dithiocarbamate hybrid 3f formed hydrogen bond interactions with Lys753 and Glu770 residues, while the O atom of the nitro group formed hydrogen bond interactions with Gly727 residues. These interactions may be the reason for the enhanced activity of aspirin-dithiocarbamate hybrid 3f on HER2 receptor, thus supporting aspirin-dithiocarbamate hybrid 3f as an ideal drug for the treatment of lung cancer.

[0140] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. An aspirin-dithiocarbamate hybrid, characterized in that The general structural formula of the aspirin-dithiocarbamate hybrid is as follows: Wherein, R is selected from aromatic groups, hydrocarbon groups or ester groups.

2. The aspirin-dithiocarbamate hybrid according to claim 1, characterized in that: The R is selected from one of the following groups:

3. A method for preparing an aspirin-dithiocarbamate hybrid, for preparing the aspirin-dithiocarbamate hybrid as claimed in claim 1, characterized in that: The following steps are involved: S1, aspirin and oxalyl chloride are subjected to chlorination reaction under the catalysis of a catalyst to obtain o-acetylsalicylic acid chloride 1; S2, piperazine, carbon disulfide and halogenated hydrocarbon react chemically under the action of the first acid binding agent to obtain dithiocarbamate 2; S3, o-acetylsalicylic acid chloride 1 and dithiocarbamate 2 are subjected to N-acylation reaction under the action of a second acid binding agent, and then separated and purified to obtain the target product aspirin-dithiocarbamate hybrid 3a-1. The synthetic route is as follows: Wherein, the definition of R is the same as that of claim 1 or 2.

4. The method for preparing aspirin-dithiocarbamate hybrids according to claim 3, characterized in that: The catalyst in step S1 is N,N-dimethylformamide.

5. The method for preparing aspirin-dithiocarbamate hybrids according to claim 3, characterized in that: The first acid binding agent in step S2 and the second acid binding agent in step S3 are both tertiary amines.

6. A pharmaceutically acceptable salt of aspirin-dithiocarbamate hybrid according to claim 1 or 2; Or a pharmaceutically acceptable salt of aspirin-dithiocarbamate hybrid obtained by the preparation method of aspirin-dithiocarbamate hybrid according to any one of claims 3 to 5.

7. A pharmaceutical composition comprising the aspirin-dithiocarbamate hybrid according to claim 1 or 2; Or a pharmaceutical composition containing the aspirin-dithiocarbamate hybrid obtained by the preparation method of the aspirin-dithiocarbamate hybrid according to any one of claims 3 to 5; Or a pharmaceutical composition containing a pharmaceutically acceptable salt of the aspirin-dithiocarbamate hybrid according to claim 6.

8. A pharmaceutical preparation containing the aspirin-dithiocarbamate hybrid according to claim 1 or 2; Or a pharmaceutical preparation containing the aspirin-dithiocarbamate hybrid obtained by the preparation method of the aspirin-dithiocarbamate hybrid according to any one of claims 3 to 5; Or a pharmaceutical preparation containing a pharmaceutically acceptable salt of the aspirin-dithiocarbamate hybrid according to claim 6, characterized in that It also includes at least one pharmaceutically acceptable excipient or carrier.

9. Use of the aspirin-dithiocarbamate hybrid according to claim 1 or 2 in the preparation of anticancer drugs; Or use of the aspirin-dithiocarbamate hybrid obtained by the method for preparing the aspirin-dithiocarbamate hybrid according to any one of claims 3 to 5 in the preparation of anticancer drugs; Or use of a pharmaceutically acceptable salt of aspirin-dithiocarbamate hybrid as claimed in claim 6 in the preparation of anticancer drugs; Or use of the pharmaceutical composition according to claim 7 in the preparation of anticancer drugs; Or use of the pharmaceutical preparation as claimed in claim 8 in the preparation of anticancer drugs.

10. Use of the aspirin-dithiocarbamate hybrid according to claim 9 in the preparation of anticancer drugs, characterized in that: The cancers that can be treated by the anticancer drug include: lung cancer, liver cancer, colorectal cancer, cervical cancer, gastric cancer and primary glioblastoma.

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