Aspirin-dithiocarbamate hybrids, methods of making and use thereof

Abstract

This invention relates to the field of aspirin medicinal chemistry, and particularly to an aspirin-dithiocarbamate hybrid, its preparation method, and its applications. The preparation method uses aspirin (ASP) as the parent compound and involves a three-step reaction: acylation, dithiocarbamate formation, and N-acylation. During this reaction, several novel aspirin-dithiocarbamate hybrids (3a–l) can be efficiently synthesized through a single silica gel column chromatography separation and purification step, making the preparation process simple. The prepared aspirin-dithiocarbamate hybrids significantly enhance the toxicity of human lung cancer cells A549, approximately eight times that of the parent compound ASP, and are comparable to the anticancer drug irinotecan. The prepared aspirin-dithiocarbamate hybrid 3f exhibits inhibitory effects on various cancer cells and can inhibit the growth of human lung cancer A549 cells by inducing cell cycle arrest in the G0 / G1 phase.

Description

Technical Field

[0001] This invention relates to the field of aspirin pharmaceutical chemistry, and more particularly to an aspirin-dithiocarbamate hybrid, its preparation method, and its application. Background Technology

[0002] Data released by the International Agency for Research on Cancer (IARC), a branch of the World Health Organization, shows that approximately 20 million new cancer cases were diagnosed globally in 2022. Lung cancer, breast cancer in women, and colorectal cancer accounted for the top three in terms of the number of new cancer cases. Currently, with the advancement of cancer treatment, a large number of patients have developed drug resistance. This is because existing chemotherapy drugs or targeted drugs directly inhibit key processes such as cancer cell proliferation signaling pathways and DNA synthesis, thereby interfering with the core proliferation mechanism of cancer cells. Cancer cells are prone to developing drug resistance by altering target structures through gene mutations and activating bypass signals. Therefore, it is necessary to develop novel anticancer drugs that exhibit high selectivity for tumor cells and do not induce drug resistance.

[0003] Aspirin is a commonly used nonsteroidal anti-inflammatory drug (NSAID) generally used to relieve pain, reduce fever, and inhibit platelet aggregation. In recent years, aspirin has also been found to have certain anti-tumor activity. Its mechanism is to reduce the synthesis of inflammatory mediators such as 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 does not cause cancer cells to develop drug resistance like chemotherapy drugs or targeted drugs. However, aspirin has limited cytotoxicity to cancer cells and at present, it can only play an adjunctive role in the treatment of cancer and cannot be considered a new type of anti-cancer drug. Summary of the Invention

[0004] In view of this, it is necessary to provide an aspirin-dithiocarbamate hybrid, its preparation method and application, which can enhance the cytotoxicity of aspirin against cancer cells by introducing a highly compatible and active fragment into aspirin, thus enabling it to become a novel anticancer drug.

[0005] In a first aspect, the present invention provides an aspirin-dithiocarbamate hybrid, the general structural formula of which is as follows:

[0006]

[0007] R is selected from aromatic, hydrocarbon, 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, for preparing the aspirin-dithiocarbamate hybrid as described in the first aspect, comprising the following steps:

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

[0012] S2, piperazine, carbon disulfide and haloalkanes 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 undergo N-acylation reaction under the action of a second acid-binding agent, followed by separation and purification to obtain the target product aspirin-dithiocarbamate hybrid 3a-l. The synthetic route is as follows:

[0014]

[0015]

[0016] The definition of R is the same as 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] Thirdly, the present invention provides applications of aspirin-dithiocarbamate hybrids as described in the first aspect, including:

[0020] Pharmaceutically acceptable salts of aspirin-dithiocarbamate hybrids as described above.

[0021] Pharmaceutical compositions of aspirin-dithiocarbamate hybrids or pharmaceutically acceptable salts thereof, as described above.

[0022] Pharmaceutical formulations of aspirin-dithiocarbamate hybrids or pharmaceutically acceptable salts thereof, as described above, also include at least one pharmaceutically acceptable excipient or carrier.

[0023] The use of the aspirin-dithiocarbamate hybrid, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a pharmaceutical preparation thereof, as described above, in the preparation of anticancer drugs.

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

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0026] 1. The aspirin-dithiocarbamate hybrid (IC) prepared by this invention 50 =26.01-132.13 μM) showed superior cytotoxicity against human lung cancer cells A549 compared to parent aspirin (IC50, 132.13 μM). 50 =202.31 μM), of which aspirin-dithiocarbamate hybrid 3b (IC50) 50 =26.01 μM) and aspirin-dithiocarbamate hybrid 3c (IC50) 50 =26.54 μM) showed the strongest cytotoxicity against human lung cancer cells A549, approximately equivalent to that of parent aspirin (IC50). 50 =202.31μM) is 8 times that of the anticancer drug irinotecan (IRT, IC50). 50 Since the concentration of aspirin-dithiocarbamate hybrid prepared in this invention is close to 12.36 μM, it can effectively treat lung cancer.

[0027] 2. The method for preparing aspirin-dithiocarbamate hybrids provided by the present invention can use aspirin (ASP) as a parent compound and proceed through three consecutive steps of acylation, dithiocarbamate formation, and N-acylation. In this reaction process, only one separation and purification is required to efficiently synthesize several novel aspirin-dithiocarbamate hybrids, and the preparation steps are simple.

[0028] 3. The aspirin-dithiocarbamate hybrid 3f prepared in this invention showed good cytotoxicity against human hepatocellular carcinoma cells HepG2, human colorectal carcinoma cells HCT-116, human colorectal carcinoma cells SW-620, human cervical carcinoma cells HeLa, human gastric carcinoma cells HGC-27, and human primary glioblastoma cells U87 (HepG2, HCT-116, SW-620, HeLa, HGC-27, U87; IC50). 50 =48.28-142.10 μM), therefore, the aspirin-dithiocarbamate hybrid prepared in this invention has a certain inhibitory effect on a variety of cancer cells;

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

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

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

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

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

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

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

[0036] Figure 7 This application relates to aspirin-dithiocarbamate hybrid 3c. 1 HNMR spectrum.

[0037] Figure 8 This application relates to aspirin-dithiocarbamate hybrid 3c. 13 CNMR spectrum.

[0038] Figure 9 This is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3c of this application.

[0039] Figure 10 This application relates to the aspirin-dithiocarbamate hybrid 3d. 1 HNMR spectrum.

[0040] Figure 11 This application relates to the aspirin-dithiocarbamate hybrid 3d.13 CNMR spectrum.

[0041] Figure 12 This application relates to the aspirin-dithiocarbamate hybrid 3d. 19 FNMR spectrum.

[0042] Figure 13 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3d of this application.

[0043] Figure 14 This application relates to the aspirin-dithiocarbamate hybrid 3e. 1 HNMR spectrum.

[0044] Figure 15 This application relates to the aspirin-dithiocarbamate hybrid 3e. 13 CNMR spectrum.

[0045] Figure 16 This application relates to the aspirin-dithiocarbamate hybrid 3e. 19 FNMR spectrum.

[0046] Figure 17 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3e of this application.

[0047] Figure 18 This application relates to the aspirin-dithiocarbamate hybrid 3f. 1 HNMR spectrum.

[0048] Figure 19 This application relates to the aspirin-dithiocarbamate hybrid 3f. 13 CNMR spectrum.

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

[0050] Figure 21 This application contains 3g of aspirin-dithiocarbamate hybrid. 1 HNMR spectrum.

[0051] Figure 22 This application contains 3g of aspirin-dithiocarbamate hybrid. 13 CNMR spectrum.

[0052] Figure 23 This application contains 3g of aspirin-dithiocarbamate hybrid. 19 FNMR spectrum.

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

[0054] Figure 25 This application relates to the aspirin-dithiocarbamate hybrid 3h. 1 HNMR spectrum.

[0055] Figure 26 This application relates to the aspirin-dithiocarbamate hybrid 3h. 13 CNMR spectrum.

[0056] Figure 27 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid of this application over 3 hours.

[0057] Figure 28 This application relates to the aspirin-dithiocarbamate hybrid 3i. 1 HNMR spectrum.

[0058] Figure 29 This application relates to the aspirin-dithiocarbamate hybrid 3i. 13 CNMR spectrum.

[0059] Figure 30 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3i of this application.

[0060] Figure 31 This application relates to aspirin-dithiocarbamate hybrid 3j. 1 HNMR spectrum.

[0061] Figure 32 This application relates to aspirin-dithiocarbamate hybrid 3j. 13 CNMR spectrum.

[0062] Figure 33 This is the HRMS spectrum of aspirin-dithiocarbamate hybrid 3j of this application.

[0063] Figure 34 This application relates to the aspirin-dithiocarbamate hybrid 3k. 1 HNMR spectrum.

[0064] Figure 35 This application relates to the aspirin-dithiocarbamate hybrid 3k. 13 CNMR spectrum.

[0065] Figure 36 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3k of this application.

[0066] Figure 37 This application relates to the aspirin-dithiocarbamate hybrid 3l. 1 HNMR spectrum.

[0067] Figure 38 This application relates to the aspirin-dithiocarbamate hybrid 3l. 13 CNMR spectrum.

[0068] Figure 39 This is the HRMS spectrum of the aspirin-dithiocarbamate hybrid 3l of this application.

[0069] Figure 40 This is a graph showing the cell inhibition rates of the aspirin-dithiocarbamate hybrid and ASP of this application on human lung cancer cells A549 and normal human liver cells LO2.

[0070] Figure 41 This is a diagram showing the apoptosis effect of the aspirin-dithiocarbamate hybrid 3f of this application on human lung cancer cells A549.

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

[0072] Figure 43 This is a model diagram of the 3f molecular docking of the aspirin-dithiocarbamate hybrid of this application. Detailed Implementation

[0073] The technical solutions and effects of the embodiments of the present invention will be further described in detail below with reference to the accompanying drawings.

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

[0075]

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

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

[0078]

[0079] The aspirin-dithiocarbamate hybrid (IC) prepared in this invention 50 =26.01-132.13 μM) showed superior cytotoxicity against human lung cancer cells A549 compared to parent aspirin (IC50, 132.13 μM). 50=202.31 μM), of which aspirin-dithiocarbamate hybrid 3b (IC50) 50 =26.01 μM) and aspirin-dithiocarbamate hybrid 3c (IC50) 50 =26.54 μM) showed the strongest cytotoxicity against human lung cancer cells A549, approximately equivalent to that of parent aspirin (IC50). 50 =202.31μM) is 8 times that of the anticancer drug irinotecan (IRT, IC50). 50 Since the concentration of aspirin-dithiocarbamate hybrid prepared in this invention is close to 12.36 μM, it can effectively treat lung cancer.

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

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

[0082] S2, piperazine, carbon disulfide and haloalkanes 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 undergo N-acylation reaction under the action of a second acid-binding agent, followed by separation and purification to obtain the target product aspirin-dithiocarbamate hybrid 3a-l. The synthetic route is as follows:

[0084]

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

[0086] Furthermore, the catalyst in step S1 is N,N-dimethylformamide; the reaction temperature in step S1 is 10℃~40℃; and the reaction time in step S1 is 0.5h~3h.

[0087] Furthermore, the first acid-binding agent in step S2 and the second acid-binding agent in step S3 are both tertiary amines, such as trimethylamine, triethylamine, and tripropylamine; the reaction temperature required in step S2 is 10℃~40℃; and the reaction time required in step S2 is 0.5h~3h.

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

[0089] The method for preparing aspirin-dithiocarbamate hybrids provided by this invention can use aspirin (ASP) as a parent compound and proceed through three consecutive steps of acylation, dithiocarbamate formation, and N-acylation. In this reaction process, only one separation and purification is required to efficiently synthesize several novel aspirin-dithiocarbamate hybrids, and the preparation steps are simple.

[0090] Thirdly, the present invention provides applications of aspirin-dithiocarbamate hybrids as described in the first aspect, including:

[0091] Pharmaceutically acceptable salts of aspirin-dithiocarbamate hybrids as described above.

[0092] Pharmaceutical compositions of aspirin-dithiocarbamate hybrids or pharmaceutically acceptable salts thereof, as described above.

[0093] Pharmaceutical formulations of aspirin-dithiocarbamate hybrids or pharmaceutically acceptable salts thereof, as described above, further include at least one pharmaceutically acceptable excipient or carrier; said pharmaceutical formulation includes tablets, powders, capsules, granules, or injections.

[0094] The use of the aspirin-dithiocarbamate hybrid, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a pharmaceutical preparation thereof, as described above, in the preparation of anticancer drugs.

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

[0096] Example 1: The general method for synthesizing aspirin-dithiocarbamate hybrids is as follows:

[0097] Experimental materials: Commercial reagents were purchased from Adamas and Energie; unless otherwise specified, they were used as received. Redistilled dichloromethane (DCM) was used. Thin-layer chromatography (TLC) was performed on silica GF254 plates. Infrared spectra were recorded on an FTIR-8400S spectrometer using KBr pellets. Results were obtained using a Bruker Avance III 400MHz spectrometer. 1 HNMR and 13 C10 NMR spectroscopy. Chemical shift (δ) is expressed in ppm with tetramethylsilane as a reference. Residual solvent signal is used as... 1 HNMR and 13 Reference for CNMR spectrum (CDCl3:δ) H =7.26ppm,δ C=77.16 ppm). High-resolution mass spectrometry (HRMS) of the target compound was performed on a Thermo Fisher LTQ Orbitrap XL. Bioactivity assays were conducted by the Collaborative Innovation Center for Green Pharmaceuticals, Zhejiang University of Technology.

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

[0099] In S201, at 25℃, piperazine (0.4 g, 4.5 mmol), DCM (10 mL), and triethylamine (0.9 mL, 6.0 mmol) were added sequentially to a 50 mL flask. A solution of CS2 (0.2 mL, 3.3 mmol) dissolved in DCM (10 mL) was slowly added dropwise. The mixture was stirred for 1 h, and then 3.3 mmol of haloalkanes RX was added. The reaction was monitored by TLC (reaction time approximately 1.5 h). 50 mL of water was added to the reaction solution, and the mixture was washed with saturated saline (30 mL x 3). The solution was extracted with DCM (30 mL x 3), and the organic phase was dried over anhydrous Na2SO4. The mixture was filtered and concentrated to remove excess solvent to obtain crude product 2.

[0100] In S301, at 0℃, crude product 2, DCM (10 mL), and triethylamine (0.9 mL, 6.0 mmol) were added sequentially to a 50 mL flask. Then, a 10 mL solution of crude product 1 in DCM was slowly added dropwise. The mixture was stirred at room temperature, and the reaction was monitored by TLC (stirring time approximately 1 h). 30 mL of water was added to the reaction mixture, and the mixture was washed with saturated saline solution (30 mL x 3). The mixture was extracted with DCM (30 mL x 3), and the organic phase was dried over anhydrous Na2SO4. The mixture was filtered, concentrated to remove excess solvent, and the residue was purified by silica gel column chromatography to obtain the target product, aspirin-dithiocarbamate hybrid 3.

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

[0102] The target products 3 obtained from the haloalkanes RX with the above-mentioned structures are numbered sequentially as 3a, 3b, 3c, 3d, 3e, 3f, 3g, 3h, 3i, 3j, 3k and 3l; the target products 3 obtained by the above synthesis method are aspirin-dithiocarbamate hybrids (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 1F 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 1H 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; 11H NMR (400 MHz, CDCl3) δ 8.15 (d, J = 8.3 Hz, 2H), 7.55 (d, J = 8.3 Hz, 2H), 7.45 (dt, J = 8.6, 4.5 Hz, 1H), 7.28 (d, J = 4.6 Hz, 2H), 7.15 (d, J = 8.2 Hz, 1H), 4.69 (s, 2H), 4.44 - 3.72 (m, 6H), 3.50 (d, J = 5.7 Hz, 2H), 2.26 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 195.67, 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 °C; 1 1H NMR (400 MHz, CDCl3) δ 7.46 - 7.40 (m, 1H), 7.34 (dd, J = 8.3, 5.4 Hz, 2H), 7.28 (d, J = 4.5 Hz, 2H), 7.15 (d, J = 8.2 Hz, 1H), 6.98 (t, J = 8.5 Hz, 2H), 4.54 (s, 2H), 4.46 - 3.64 (m, 6H), 3.46 (t, J = 5.1 Hz, 2H), 2.25 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.73, 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 H 21 N2O3S2 [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 test of aspirin-dithiocarbamate hybrid 3

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

[0118] Experimental method: First, human lung cancer cells A549 or human normal liver cells LO2 (5×10⁻⁶) were used. 3100 μL was inoculated into 96-well plates in triplicate. After incubation at 37°C for 24 hours, the suspension was replaced with fresh medium containing different doses of aspirin-dithiocarbamate hybrid 3a-l, ASP, and IRT (0.01 μM, 0.1 μM, 1 μM, 10 μM, and 100 μM). The same volume of dimethyl sulfoxide (DMSO) was added to the negative control wells and the solvent control wells. Then, 10 μL of the solution was added to each well. 3-(4,5-Dimethylthiazolyl-2-yl)2,5-diphenyltetrazolium bromide (MTT) (5 mg / mL, phosphate-buffered saline (PBS)) was cultured for 4 hours, after which the culture medium was removed. The MTT formazan precipitate was then dissolved in 100 μL LDMSO, mechanically shaken for 10 minutes, and immediately read using a microplate reader (FlexStation3, Molecular Devices) at 570 nm. Simultaneously, the IC50 of hybrid 3a-l was calculated using GraphPad software. 50 The values ​​and results are shown in Table 1 below:

[0119] Table 1

[0120]

[0121]

[0122] Analysis of the data in the table reveals that aspirin-dithiocarbamate hybrid 3a-l (IC) 50 =26.01-55.75 μM) showed significantly higher cytotoxicity against human lung cancer cells A549 than that against the parent ASP (IC50) 50 =202.31 μM), aspirin-dithiocarbamate hybrid 3k (IC50) 50 Except for 132.13 μM. When R is 4-methylbenzyl (3b, IC 50 =26.01 μM) and 4-cyanobenzyl (3c, IC) 50 The cytotoxicity was strongest at 26.54 μM, approximately equal to the parent ASP (IC50). 50 =202.31 μM) 8 times, compared with the positive control IRT (IC 50 =12.36 μM). Aspirin-dithiocarbamate hybrid 3f (SI=6) showed the highest selectivity for human lung cancer cells (LO2 / A549), which was higher than ASP (SI=1) and IRT (SI=1).

[0123] Calculate the cell inhibition rate and plot them as follows: Figure 40 The line graph and bar chart shown indicate that the cell inhibition rate = [A570 (negative control well) - A570 (drug administration well)] / A570 (negative control well) × 100%.

[0124] from Figure 40 The data show that as the concentration of aspirin-dithiocarbamate hybrid 3a-l gradually increases, the inhibition rate against human lung cancer cells A549 gradually increases. At a concentration of 100 μM, the inhibition rates of aspirin-dithiocarbamate hybrids 3b, 3c, and 3f against human lung cancer cells A549 are as high as 92.06%, 78.32%, and 79.92%, respectively, while the inhibition rate of the parent aspirin against human lung cancer cells A549 is only 49.38%. Therefore, introducing dithiocarbamate into aspirin can significantly enhance the cytotoxicity of parent aspirin against human lung cancer cells 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 was investigated, including human hepatocellular carcinoma HepG2, human colorectal carcinoma HCT-116, human colorectal carcinoma SW-620, human cervical carcinoma HeLa, human gastric carcinoma HGC-27, and human primary glioblastoma U87. 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 is effective against the above six types of human cancer cells (ICV). 50 =48.28-142.10 μM) all showed good cytotoxicity and had certain therapeutic effects on various cancer cells; at the same time, aspirin-dithiocarbamate hybrid 3f showed good cytotoxicity against human lung cancer cells A549 (IC50). 50 The cytotoxicity of aspirin-dithiocarbamate hybrid 3f (31.08 μM) was higher than that of the six human cancer cells mentioned above, 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: First, human lung cancer A549 cells (5×10⁻⁶) were... 4Three copies (1000 μL each) were seeded into 12-well plates and cultured at 37°C for 24 h. The old medium was then replaced with fresh medium containing different concentrations of aspirin-dithiocarbamate hybrid 3f (20, 80, and 160 μM). Simultaneously, the same volume of dimethyl sulfoxide was added to the negative control wells and the solvent control wells. The cells were then cultured at 37°C for 48 h. After aspirating the medium, 100 μL of pre-warmed 0.25% trypsin without EDTA was added to each well. After trypsinization, the treated cells were stained using the Annexin V-FITC / PI apoptosis detection kit according to the manufacturer's instructions. The cells were then incubated in the dark at room temperature for 5–15 minutes, and apoptotic cells were analyzed using V-FITC / PI flow cytometry. The results are shown below. Figure 41 As shown.

[0132] right Figure 41 Analysis of Figure A shows that with increasing concentrations of aspirin-dithiocarbamate hybrid 3f, the number of early and late apoptotic cells gradually increases, while the number of normal cells gradually decreases; Figure 41 Analysis of Figure B shows that when the concentrations of aspirin-dithiocarbamate hybrid 3f were 20, 80, and 160 μM, the total apoptosis rates of human lung cancer cells A549 were 35.54%, 39.58%, and 60.70%, respectively, while the control group showed only 6.61%. These results indicate that aspirin-dithiocarbamate hybrid 3f induces apoptosis in human lung cancer A549 cells in a concentration-dependent manner.

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

[0134] Experimental method: First, human lung cancer A549 cells (2.5 × 10⁻⁶) were... 5 Three copies (2500 μL each) were seeded into 6-well plates and cultured at 37°C for 24 h. The old medium was then replaced with fresh medium containing different concentrations of aspirin-dithiocarbamate hybrid 3f (5, 10, and 20 μM). Simultaneously, the same volume of dimethyl sulfoxide was added to the negative control wells and solvent control wells. The plates were then cultured at 37°C for another 24 h. After aspirating the medium, 200 μL of 0.25% trypsin was added to each well. Following trypsin treatment, the cells were fixed overnight with 70% ethanol at 4°C. Staining was then performed 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. The results are shown below. Figure 42 As shown.

[0135] right Figure 42Analysis revealed that the number of human lung cancer A549 cells with cell cycle arrest gradually increased with increasing concentrations of aspirin-dithiocarbamate hybrid 3f. When the concentrations of aspirin-dithiocarbamate hybrid 3f were 5, 10, and 20 μM, human lung cancer A549 cells gradually arrested in the G0 / G1 phase. These results indicate that aspirin-dithiocarbamate hybrid 3f can induce cell cycle arrest in the G0 / G1 phase in human lung cancer A549 cells, thereby inhibiting their growth.

[0136] Example 6: Binding characteristics analysis of aspirin-dithiocarbamate hybrid 3f with receptor HER2

[0137] Experimental materials: Molecules were drawn using ChemDraw 20.0 software and minimized using MOE (Molecular Manipulation Environment) software. The receptor HER2 (PDBID: 3PP0) was downloaded from the protein database (https: / / www.rcsb.org) and prepared using MOE software. The pockets in HER2 were set in a solvent environment.

[0138] Experimental Methods: Molecular docking analysis was performed using the Molecular Manipulation Environment (MOE). After setting the method (placement: triangular matcher, refinement: rigid acceptor), score (placement: LondondG, refinement: GBVI / WSAdG), and pose (placement: 300, refinement: 5), docking was performed using MOE software. The docking conformation with the lowest binding energy was selected from 300 conformations as the representative binding model for the corresponding compound. Figure 43 As shown.

[0139] right Figure 43 Analysis revealed that aspirin-dithiocarbamate hybrid 3f is well-encapsulated within the active pocket of the HER2 receptor. Furthermore, the O atoms in the carbonyl and ester groups of aspirin-dithiocarbamate hybrid 3f form hydrogen bonds with Lys753 and Glu770 residues, while the O atom in the nitro group forms a hydrogen bond with Gly727 residue. These interactions may explain the enhanced HER2 receptor activity of aspirin-dithiocarbamate hybrid 3f, thus supporting its potential as an ideal drug for treating lung cancer.

[0140] The above are merely 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 within the protection scope of the present invention.

Claims

1. An aspirin-dithiocarbamate hybrid or a pharmaceutically acceptable salt thereof, characterized in that, The general structural formula of the aspirin-dithiocarbamate hybrid is as follows: ； wherein R is selected , , , , .

2. A process for the preparation of an aspirin-dithiocarbamate hybrid according to claim 1, characterized in that, Includes the following steps: S1, Aspirin and oxalyl chloride are subjected to an acyl chloride reaction under the catalysis of a catalyst to obtain o-acetylsalicylic acid chloride 1; the catalyst is N,N-dimethylformamide; S2, piperazine, carbon disulfide and haloalkanes react chemically under the action of the first acid-binding agent to obtain dithiocarbamate 2; S3, after N-acylation of o-acetylsalicylic acid chloride 1 and dithiocarbamate 2 under the action of a second acid-binding agent, the aspirin-dithiocarbamate hybrid of claim 1 is obtained by separation and purification. The synthetic route is as follows: ； ； ； The definition of R is the same as in claim 1.

3. The method for preparing the aspirin-dithiocarbamate hybrid as described in claim 2, characterized in that, The first acid-binding agent in step S2 and the second acid-binding agent in step S3 are both tertiary amines.

4. A pharmaceutical composition, characterized by, Contains the aspirin-dithiocarbamate hybrid as described in claim 1 or a pharmaceutically acceptable salt thereof.

5. A pharmaceutical preparation, characterized by, It contains the aspirin-dithiocarbamate hybrid as described in claim 1 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient or carrier.

6. The use of the aspirin-dithiocarbamate hybrid of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 4, in the preparation of an anticancer drug; wherein the cancer is selected from lung cancer, liver cancer, colorectal cancer, and cervical cancer.

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