Tripterine-platinum (IV) complex with anti-tumor activity as well as preparation method and application of tripterine-platinum (IV) complex

Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a tripterine-platinum (IV) complex with antitumor activity as well as a preparation method and application of the tripterine-platinum (IV) complex. The tripterine-platinum (IV) complex takes tripterine as an axial ligand, and a platinum center skeleton structure is cis-platinum, carboplatin or oxaliplatin. The tetravalent platinum complex can induce para-apoptosis of cancer cells, so that the common problem of drug resistance caused by apoptosis resistance of the cancer cells in tumor chemotherapy is solved. Besides, the tetravalent platinum complex has the characteristics of high efficiency and low toxicity, the in-vitro cytotoxicity of the tetravalent platinum complex to tumor cells such as non-small cell lung cancer A549 cells and cisplatin-resistant strains A549cisR thereof is higher than that of clinically applied metal anti-cancer drug cisplatin, and the tetravalent platinum complex has low toxicity to normal cells MRC-5, so that the tetravalent platinum complex has a good application prospect. Good selectivity and relatively high medicinal value are shown in the aspect of inhibiting the growth of tumor cells.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to a tripterygium wilfordii-platinum (IV) complex with anti-tumor activity and a preparation method and application thereof. Background Art

[0002] Cancer is a malignant disease that threatens human life and health. Chemotherapy is one of the main means of clinical treatment of cancer. At present, platinum anticancer drugs are one of the most widely used chemotherapy drugs in clinical practice. Platinum drugs have good anticancer effects and a broad anticancer spectrum, and are widely used to treat lung cancer, cervical cancer, ovarian cancer and other cancers. However, the drug resistance and toxic side effects of platinum drugs, especially the drug resistance problem, are still the key factors restricting their clinical effects and applications. Therefore, it is urgent to develop new platinum anticancer drugs to solve their drug resistance problem.

[0003] Taking cisplatin as an example, the main resistance mechanisms of platinum-based anticancer drugs are as follows: (1) The stage before cisplatin interacts with DNA: the uptake of cisplatin by tumor cells decreases, the amount of drug pumped out by cancer cells increases, and cisplatin is inactivated by binding to thiol-containing biological molecules; (2) The stage of interaction between cisplatin and DNA: mainly including the increased tolerance of cancer cells to Pt-DNA damage and the enhanced repair ability of cancer cells to Pt-DNA damage; (3) The stage of inducing cell apoptosis after cisplatin causes DNA damage: the downregulation of pro-apoptotic proteins and the upregulation of anti-apoptotic proteins in cancer cells will lead to apoptosis resistance in cancer cells; (4) Other pathways.

[0004] Inducing apoptosis in cancer cells is the main mechanism of action of platinum anticancer drugs to exert their anticancer activity. Cancer cell resistance to apoptosis is one of the main causes of platinum drug resistance. Therefore, synthesizing new platinum complexes to induce non-apoptotic cell death in cancer cells is an effective strategy to overcome platinum resistance. Summary of the invention

[0005] The purpose of the present invention is to provide a tripterygium wilfordii-platinum (IV) complex and a preparation method and application thereof, so as to effectively overcome the drug resistance problem of platinum drugs in tumor chemotherapy and achieve selective killing of cancer cells.

[0006] The present invention adopts the following technical solutions:

[0007] A tripterygium wilfordii-platinum (IV) complex, wherein the tripterygium wilfordii-platinum (IV) complex has a general structural formula as shown in Formula S1:

[0008]

[0009] Among them, in formula S1, R represents One of them.

[0010] The preparation method of the above-mentioned tripterygium wilfordii-platinum (IV) complex comprises the following steps:

[0011] (1) Cisplatin, carboplatin or oxaliplatin reacts with hydrogen peroxide at 50-75° C. to obtain a dihydroxy tetravalent platinum complex; the structural formula of the dihydroxy tetravalent platinum complex is as follows:

[0012]

[0013] (2) a dihydroxy tetravalent platinum complex is subjected to a condensation reaction with Fmoc-beta-alanine in the presence of a condensing agent and an organic base to obtain a tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc; the tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc has the following structural formula:

[0014]

[0015] (3) piperidine is added to the tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc obtained in step (2), respectively, and the reaction is carried out at room temperature with DMF as solvent. After the reaction is completed, the tetravalent platinum complex cisPt(IV)-mNH2, carPt( IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2; the structural formula of the tetravalent platinum complex cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2 is as follows:

[0016]

[0017] (4) The tetravalent platinum complexes cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2 obtained in step (3) are subjected to condensation reaction with the ligand tripterygium wilfordii (Cel) in the presence of a condensing agent and an organic base to obtain tetravalent platinum complexes Pt1, Pt2, Pt4, Pt5, Pt7 and Pt8 in sequence; the structures of the tetravalent platinum complexes Pt1, Pt2, Pt4, Pt5, Pt7 and Pt8 are as follows:

[0018]

[0019]

[0020] (5) The tetravalent platinum complexes Pt1, Pt4, and Pt7 obtained in step (4) are subjected to condensation reaction with 4-formylphenoxyacetic acid (4-FP) in the presence of a condensing agent and an organic base to obtain tetravalent platinum complexes Pt3, Pt6, and Pt9, respectively; the structural formulas of the tetravalent platinum complexes Pt3, Pt6, and Pt9 are as follows:

[0021]

[0022] Furthermore, the condensing agent in steps (2), (4) and (5) is any one of EDCi, DCC, DIC, HATU or HBTU, or a combination of any one of EDCi, DCC, DIC, HATU or HBTU and any one of NHS, HOBt or HOAt; and the organic base is any one of DMF, TEA or DIPEA.

[0023] Furthermore, in step (2), the reaction equivalent ratio of the dihydroxy tetravalent platinum complex to Fmoc-beta-alanine, the condensation agent, and the organic base is 0.2-1:1.2:1.4:0.1.

[0024] Furthermore, the solvent for the condensation reaction in steps (2), (4) and (5) is DMSO, the reaction temperature is 20 to 50° C., the reaction time is 24 to 72 hours, and the product is purified by column chromatography.

[0025] Furthermore, in step (2), the post-treatment method of the obtained product is: filtering to remove insoluble matter, adding an equal amount of DCM to the solution obtained by filtration, mixing, and dripping into ether, standing, and centrifuging to separate the precipitate; the product is further purified by column chromatography, and the eluent is a mixture of dichloromethane (DCM) and methanol in a volume ratio of 6 to 16:1.

[0026] Furthermore, in step (3), the reaction equivalent ratio of the tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc to piperidine is 1:13 to 26.

[0027] Furthermore, the reaction temperature of the reaction in step (3) is 20-40° C., the reaction time is 0.5-3 hours, and after the reaction is completed, the insoluble matter is filtered out, the clear liquid is dripped into ether and allowed to stand; the white precipitate is collected by centrifugation, the precipitate is washed with ether, and vacuum dried.

[0028] Furthermore, in step (4), the reaction equivalent ratio of the tetravalent platinum complex cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2 with Cel, condensation agent and organic base is 0.2-1:1.2:1.4:0.1.

[0029] Furthermore, in step (5), the reaction equivalent ratio of the tetravalent platinum complex Pt1, Pt4 or Pt7 to 4-formylphenoxyacetic acid, condensation agent and organic base is 0.2-1:1.2:1.4:0.1.

[0030] Furthermore, in step (5), the post-treatment method of the obtained product is: filtering to remove insoluble matter, adding an equal amount of DCM to the solution and dripping it into ether, standing, and centrifuging to separate the precipitate; further purifying the product by column chromatography, the eluent is a mixture of dichloromethane (DCM) and methanol, the volume ratio of which is 10 to 16:1.

[0031] The present invention also provides the use of the tripterygium wilfordii-platinum (IV) complex in the preparation of anti-tumor drugs.

[0032] Preferably, the anti-tumor drug uses the tetravalent platinum complex of the present invention as the sole active ingredient or is combined with other anti-tumor active ingredients, and is allowed to include pharmaceutically acceptable excipients.

[0033] Preferably, the tripterygium wilfordii-platinum (IV) complex is one of the following structures:

[0034]

[0035] Preferably, the tumor is human non-small cell lung cancer cell A549 or human non-small cell lung cancer cell cisplatin-resistant strain A549cisR.

[0036] The beneficial effects of the present invention are:

[0037] The present invention provides a tripterine-platinum (IV) complex, wherein the tripterine-platinum (IV) complex has tripterine as an axial ligand, and the platinum center skeleton structure is cisplatin, carboplatin or oxaliplatin. This type of tetravalent platinum complex can induce cancer cells to undergo secondary apoptosis, which is manifested as cytoplasmic vacuolation, mainly endoplasmic reticulum and mitochondrial expansion. By inducing cancer cells to undergo secondary apoptosis, the drug resistance problem caused by cancer cells producing apoptosis resistance in tumor chemotherapy is improved.

[0038] In addition, this tetravalent platinum complex has the characteristics of high efficiency and low toxicity. Not only is its in vitro cytotoxicity to tumor cells such as non-small cell lung cancer A549 cells and its cisplatin-resistant strain A549cisR higher than that of the clinically used anticancer drug cisplatin, but it also has less killing effect on normal cells MRC-5. It shows good selectivity and high medicinal value in inhibiting the growth of tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is the H NMR spectrum of Pt1.

[0040] Figure 2 This is the mass spectrum of Pt1.

[0041] Figure 3 This is the H NMR spectrum of Pt2.

[0042] Figure 4 This is the mass spectrum of Pt2.

[0043] Figure 5 This is the H NMR spectrum of Pt3.

[0044] Figure 6 This is the mass spectrum of Pt3.

[0045] Figure 7 This is the H NMR spectrum of Pt4.

[0046] Figure 8 This is the mass spectrum of Pt4.

[0047] Fig. 9 This is the H NMR spectrum of Pt5.

[0048] Fig.10 This is the mass spectrum of Pt5.

[0049] Fig.11 This is the H NMR spectrum of Pt6.

[0050] Fig.12 This is the mass spectrum of Pt6.

[0051] Fig.13 This is the H NMR spectrum of Pt7.

[0052] Fig.14 This is the mass spectrum of Pt7.

[0053] Fig.15 This is the H NMR spectrum of Pt8.

[0054] Fig.16 This is the mass spectrum of Pt8.

[0055] Fig.17 This is the H NMR spectrum of Pt9.

[0056] Fig.18 This is the mass spectrum of Pt9.

[0057] Fig.19 This is a picture of the complex Pt2 inducing cavitation in cancer cells. DETAILED DESCRIPTION

[0058] In order to more clearly illustrate the purpose, technical solutions and advantages of the embodiments of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and embodiments. The described embodiments are partial embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without paying creativity also belong to the protection scope of the present invention.

[0059] The abbreviations involved in the examples of the present invention include: cisplatin (cDDP), carboplatin, oxaliplatin, celastrol (Cel), carbodiimide hydrochloride (EDCi), 1,3-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), 2-(7-azabenzotriazole)-N,N,N,N'-tetramethyluronium hexafluorophosphate ( HATU), O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU), 1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azobenzotriazole (HOAt), N-hydroxysuccinimide (NHS), N,N-dimethylformamide (DMF), triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), dimethyl sulfoxide (DMSO), dichloromethane (DCM), hydrogen peroxide (H2O2), water (H2O).

[0060] There is no particular limitation on the administration method of the drug of the present invention, and the commonly used method in the art can be used. The drug can be administered by injection or oral administration. The drug of the present invention can be prepared into various dosage forms corresponding to the administration method used, and the frequency of administration is not particularly limited, and the drug can be administered once or multiple times a day.

[0061] Example 1 Synthesis of cisPt(IV), carPt(IV) and oxaPt(IV)

[0062] Synthesis of cisPt(IV):

[0063]

[0064] Take 500.0 mg of cisplatin (cDDP, 1.7 mmol), add 15.0 mL of H2O, then add 2.5 mL of 30% H2O2, stir at 65°C in the dark for 4-8 hours. After the reaction is completed, cool at 4°C overnight, and a yellow precipitate will precipitate. Filter, wash and dry to obtain 430.3 mg of light yellow solid, with a yield of 80.5%.

[0065] Synthesis of carPt(IV):

[0066]

[0067] The preparation method is the same as cisPt(IV), except that cisplatin is replaced by carboplatin (1.3 mmol), and 450.3 mg of white solid is obtained with a yield of 82.5%.

[0068] Synthesis of oxaPt(IV):

[0069]

[0070] The preparation method is the same as cisPt(IV), except that cisplatin is replaced by oxaliplatin (1.3 mmol), and 451.6 mg of white solid is obtained with a yield of 83.2%.

[0071] Example 2 Synthesis of cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc and oxaPt(IV)-diFmoc

[0072] Synthesis of cisPt(IV)-mFmoc:

[0073]

[0074] CisPt(IV) (200.0 mg, 0.6 mmol), Fmoc-beta-alanine (62.0 mg, 0.2 mmol), condensing agent (0.2 mmol), and organic base (0.02 mmol) were added to DMSO solvent and reacted for 24 h at room temperature in the dark. After the reaction, the insoluble matter in the reaction system was filtered out, and an equal amount of DCM was added to the solution. The solution was dropped into ether solution, allowed to stand, and centrifuged to obtain a light yellow precipitate, which was dried to obtain 106.4 mg of a light yellow solid powder with a yield of 84.8%. 1 H NMR (600MHz, DMSO-d6) δ: 7.89 (d, J = 7.4Hz, 2H), 7.69 (d, J = 7.4Hz, 2H), 7.41 (t, J = 7.4Hz, 2H), 7.33 (t, J = 7.4Hz, 2H), 7.12 (t, J = 5.8 Hz,1H),6.09–5.77(m,6H),4.29(d,J=7.0Hz,2H),4.21(t,J=7.0Hz,1H),3.17(q,J=6.8Hz,2H),2.30(t,J=7.0Hz,2H),0.95(s,1H).

[0075] Synthesis of carPt(IV)-Fmoc:

[0076]

[0077] The preparation method is the same as cisPt(IV)-mFmoc, except that cisPt(IV) (200.0 mg, 0.6 mmol) is replaced by carPt(IV) (243.0 mg, 0.6 mmol) to obtain 105.5 mg of white solid powder with a yield of 75.5%. 1 H NMR(600MHz,DMSO-d6)δ:7.88(d,J=7.4Hz,2H),7.68(d,J=7.4Hz,2H),7.41(t, J=7.4Hz,2H),7.33(t,J=7.4Hz,2H),7.16(t,J=5.6Hz,1H),6.05–5.71(m,6H), 4.30(d,J=6.8Hz,2H),4.20(t,J=6.8Hz,1H),3.16(q,J=6.8Hz,2H),2.56(t,J= 8.2Hz, 2H), 2.47 (t, J = 8.2Hz, 2H), 2.31 (t, J = 7.2Hz, 2H), 1.79 (p, J = 7.8Hz, 2H).

[0078] Synthesis of oxaPt(IV)-mFmoc:

[0079]

[0080] The preparation method is the same as that of cisPt(IV)-Fmoc, except that cisPt(IV) (200.0 mg, 0.6 mmol) is replaced by oxaPt(IV) (259.0 mg, 0.6 mmol), and 99.2 mg of white solid powder is obtained with a yield of 68.4%. 1 H NMR(400MHz, DMSO-d6)δ:8.33(t,J=8.6Hz,1H),8.12(d,J=8.6Hz,1H),7.89(d,J=7.4Hz,2H),7 .85(d,J=8.0Hz,1H),7.67(d,J=7.4Hz,2H),7.41(t,J=7.4Hz,2H),7.33(t,J=7.4Hz,2H),7.24( t,J=5.8Hz,1H),7.12(t,J=9.8Hz,1H),4.28(d,J=6.8Hz,2H),4.20(t,J=6.8Hz,1H),3.14(q,J= 7.0Hz,2H),2.34(t,J=6.8Hz,2H),2.05(t,J=6.8Hz,2H),1.54–1.31(m,5H),1.15–1.04(m,3H).

[0081] Synthesis of cisPt(IV)-diFmoc:

[0082]

[0083] cisPt(IV) (36.0 mg, 0.1 mmol), Fmoc-beta-alanine (200.0 mg, 0.6 mmol), condensing agent (0.6 mmol), and organic base (0.01 mmol) were added to DMSO solvent and reacted at 50°C in the dark for 24 h. After the reaction, the insoluble matter in the reaction system was removed by filtration, and an equal amount of DCM was added to the solution and dripped into the ether solution, and the solution was allowed to stand, and a white precipitate was obtained by centrifugation, and 68.0 mg of white solid powder was obtained by drying, with a yield of 73.8%. 1 H NMR(400MHz, DMSO-d6)δ:7.89(d,J=7.4Hz,4H),7.68(d,J=7.4Hz,4H),7.41(t,J=7.4Hz,4H),7.33(t,J=7.4Hz,4H),7.18 (t,J=5.8Hz,2H),6.49(s,6H),4.31(d,J=6.8Hz,4H),4.21(t,J=6.8Hz,2H),3.17(q,J=6.8Hz,4H),2.37(t,J=7.2Hz,4H).

[0084] Synthesis of carPt(IV)-diFmoc:

[0085]

[0086] The preparation method is the same as cisPt(IV)-diFmoc, except that cisPt(IV) (36.0 mg, 0.1 mmol) is replaced by carPt(IV) (43.0 mg, 0.1 mmol) to obtain 67.5 mg of white solid powder with a yield of 68.0%. 1 H NMR(400MHz,DMSO-d6)δ:7.88(d,J=7.4Hz,4H),7.68(d,J=7.4Hz,4H),7.40(t ,J=7.4Hz,4H),7.33(t,J=7.4Hz,4H),7.16(t,J=5.6Hz,2H),6.28(s,6H),4.3 0(d,J=6.8Hz,4H),4.20(t,J=6.8Hz,2H),3.17(q,J=6.8Hz,4H),2.64(t,J=8. 2Hz, 2H), 2.47 (t, J = 8.2Hz, 2H), 2.37 (t, J = 7.2Hz, 4H), 1.79 (p, J = 7.8Hz, 2H). Synthesis of oxaPt(IV)-diFmoc:

[0087]

[0088] The preparation method is the same as that of cisPt(IV)-diFmoc, except that cisPt(IV) (36.0 mg, 0.1 mmol) is replaced by oxaPt(IV) (46.0 mg, 0.1 mmol) to obtain 39.5 mg of white solid powder with a yield of 38.8%. 1 H NMR(400MHz, DMSO-d6)δ:8.34(s,2H),8.04(s,2H),7.89(d,J=7.4Hz,4H),7.67(d,J=7.3Hz,4H),7.41(t,J=7.4Hz,4H),7.35–7.28(m,6H),4 .28(t,J=8.0Hz,4H),4.20(t,J=6.8Hz,2H),3.17(q,J=6.8Hz,4H),2.40(t,J=6.8Hz,4H),2.07–2.22(m,4H),1.45(s,4H),1.10–1.03(m,2H).

[0089] Example 3 Synthesis of cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 and oxaPt(IV)-diNH2

[0090] Synthesis of cisPt(IV)-mNH2:

[0091]

[0092] cisPt(IV)-mFmoc (70.0 mg, 0.1 mmol) and piperidine (127 μL, 1.3 mmol) were added to DMF solvent and reacted for 0.5 to 3 h at room temperature in the dark. After the reaction, the reaction solution was concentrated by rotary evaporation, dripped into ether solution, allowed to stand, centrifuged to obtain a white precipitate, and dried to obtain 35.6 mg of a white solid powder with a yield of 87.9%. 1 H NMR (400MHz, DMSO-d6) δ: 5.97 (s, 6H), 2.66-2.69 (m, 2H), 2.29 (t, J = 6.7Hz, 2H).

[0093] Synthesis of carPt(IV)-mNH2:

[0094]

[0095] The preparation method is the same as that of cisPt(IV)-mNH2, except that cisPt(IV)-mFmoc (70.0 mg, 0.1 mmol) is used to replace carPt(IV)-mFmoc (70.0 mg, 0.1 mmol) to obtain 39.4 mg of white solid powder with a yield of 82.7%. 1 H NMR (400MHz, DMSO-d6) δ: 5.94 (s, 6H), 2.71 (m, 2H), 2.56 (t, J = 6.6Hz, 2H), 2.49 (t, J = 6.6Hz, 2H), 2.29 (t, J = 6.6Hz, 2H), 1.80 (p, J = 6.7Hz, 2H).

[0096] Synthesis of oxaPt(IV)-mNH2:

[0097]

[0098] The preparation method is the same as that of cisPt(IV)-mNH2, except that cisPt(IV)-mFmoc (70.0 mg, 0.1 mmol) is used to replace oxaPt(IV)-mFmoc (72.0 mg, 0.1 mmol), and 41.3 mg of white solid powder is obtained with a yield of 82.2%. 1 H NMR (400MHz, DMSO-d6) δ: 2.73–2.66 (m, 2H), 2.29 (t, J = 6.7Hz, 2H), 2.09 (t, J = 6.8Hz, 2H), 1.57–1.36 (m, 5H), 1.21–1.06 (m, 3H).

[0099] Synthesis of cisPt(IV)-diNH2:

[0100]

[0101] cisPt(IV)-diFmoc (50.0 mg, 0.054 mmol) and piperidine (137 μL, 1.4 mmol) were added to DMF solvent and reacted for 0.5 to 3 h at room temperature in the dark. After the reaction, the reaction solution was concentrated by rotary evaporation, added dropwise to ether solution, allowed to stand, and centrifuged to obtain a white precipitate, which was dried to obtain 18.6 mg of a white solid with a yield of 72.3%. 1 H NMR (400MHz, DMSO-d6) δ: 6.43 (s, 6H), 2.73–2.66 (m, 4H), 2.35 (t, J = 6.6Hz, 4H).

[0102] Synthesis of carPt(IV)-diNH2:

[0103]

[0104] The preparation method is the same as that of cisPt(IV)-diNH2, except that cisPt(IV)-diFmoc (50.0 mg, 0.054 mmol) is used to replace carPt(IV)-diFmoc (53.6 mg, 0.054 mmol), and 20.8 mg of white solid is obtained with a yield of 70.4%. 1 H NMR (400MHz, DMSO-d6) δ: 6.25 (s, 6H), 2.71 (m, 4H), 2.62 (t, J = 6.6Hz, 2H), 2.49 (t, J = 6.6Hz, 2H), 2.29 (t, J = 6.6Hz, 4H), 1.80 (p, J = 6.7Hz, 2H).

[0105] Synthesis of oxaPt(IV)-diNH2:

[0106]

[0107] The preparation method is the same as that of cisPt(IV)-diNH2, except that cisPt(IV)-diFmoc (50.0 mg, 0.054 mmol) is used to replace oxaPt(IV)-diFmoc (55.0 mg, 0.054 mmol), and 20.5 mg of white solid is obtained with a yield of 66.2%. 1 H NMR (400MHz, DMSO-d6) δ: 2.73–2.66 (m, 4H), 2.29 (t, J = 6.7Hz, 4H), 2.09 (t, J = 6.8Hz, 2H), 1.57–1.36 (m, 5H), 1.21–1.06 (m, 3H).

[0108] Example 4 Synthesis of tetravalent platinum complex Pt1

[0109]

[0110] cisPt(IV)-mNH2 (38.8 mg, 0.148 mmol), Cel (200.0 mg, 0.444 mmol), condensing agent (0.148 mmol), and organic base (0.015 mmol) were added to DMSO solvent and reacted for 24 h at room temperature in the dark. After the reaction, the insoluble matter in the reaction system was filtered out, and an equal amount of DCM was added to the clear solution obtained by filtration, and then dripped into ether solution, allowed to stand, and centrifuged to obtain a red precipitate, which was dried to obtain 91.0 mg of a red solid powder with a yield of 73.4%.

[0111] 1 H NMR (400MHz, DMSO-d6) δ: 8.73 (s, 1H), 7.46 (t, J = 5.8Hz, 1H), 7.08 (d, J = 7.0Hz, 1H), 6.41 (s, 1H), 6.3 5(d,J=7.2Hz,1H),6.20–5.59(m,6H),3.24–3.00(m,2H),2.40(d,J=14.8Hz,1H),2.24–2.14(m,3H),2 .09(s,3H),2.07–1.91(m,2H),1.87–1.74(m,1H),1.68–1.48(m,7H),1.47–1.40(m,1H),1.38(s,3H) ,1.29(d,J=16.0Hz,1H),1.21(s,3H),1.07(s,3H),1.03(s,3H),0.87(d,J=15.0Hz,1H),0.53(s,3H)( Figure 1). HRMS (ESI) m / z [MH] - calculated for C 32 H 49 Cl2N3O6Pt:836.2554; Found:836.2494( Figure 2 ).

[0112] Example 5 Synthesis of tetravalent platinum complex Pt2

[0113]

[0114] cisPt(IV)-diNH2 (35.2 mg, 0.074 mmol), Cel (200.0 mg, 0.444 mmol), condensing agent (0.148 mmol), and organic base (0.015 mmol) were added to DMSO solvent and reacted at 50°C in the dark for 24 hours. After the reaction, the insoluble matter in the reaction system was filtered out, and the clear liquid was added with an equal amount of DCM and dripped into the ether solution, allowed to stand, and centrifuged to obtain a red precipitate, which was dried to obtain 64.6 mg of a red solid powder with a yield of 65.0%.

[0115] 1 H NMR(400MHz, DMSO-d6)δ:8.72(s,2H),7.45(s,2H),7.08(d,J=6.8Hz,2H),6.47(s,6H),6.40(s ,2H),6.35(d,J=7.0Hz,2H),3.25–3.01(m,4H),2.40(d,J=14.6Hz,2H),2.29–2.16(m,6H),2.09 (s,6H),2.07–1.72(m,6H),1.68–1.48(m,14H),1.43(d,J=15.6Hz,2H),1.38(s,6H),1.30(d,J= 15.4Hz,2H),1.21(s,6H),1.07(s,6H),1.04(s,6H),0.87(d,J=12.6Hz,2H),0.63–0.43(m,6H)( Figure 3 ). HRMS (ESI) m / z [MH] - calculated for C 64 H 90 Cl2N4O 10 Pt:1339.5599; Found:1339.5481( Figure 4 ).

[0116] Example 6 Synthesis of tetravalent platinum complex Pt3

[0117]

[0118] Pt1 (30.0 mg, 0.036 mmol), 4-formylphenoxyacetic acid (39.0 mg, 0.216 mmol), condensing agent (0.036 mmol), and organic base (0.004 mmol) were added to DMSO solvent and reacted at 50°C in the dark for 24 hours. After the reaction was completed, the insoluble matter in the reaction system was filtered out, and an equal amount of DCM was added to the obtained clear liquid, and then dripped into the ether solution, allowed to stand, and centrifuged to obtain a reddish brown precipitate, which was dried to obtain 28.0 mg of a reddish brown solid powder with a yield of 77.8%.

[0119] 1 H NMR (400MHz, DMSO-d6) δ: 9.86 (s, 1H), 8.74 (s, 1H), 7.82 (d, J = 8.6Hz, 2H), 7.53 (t, J = 5.0Hz, 1H), 7.09 (m, 3H), 6.54(s,6H),6.41(s,1H),6.35(d,J=7.0Hz,1H),4.75(s,2H),3.24–3.00(m,2H),2.40(d,J=14.8Hz,1H),2.30- 2.17(m,3H),2.09(s,3H),2.04–1.91(m,2H),1.88–1.76(m,1H),1.69–1.48(m,7H),1.43(d,J=12.0Hz,1H),1. 38(s,3H),1.29(d,J=13.0Hz,1H),1.21(s,3H),1.07(s,3H),1.04(s,3H),0.87(d,J=11.2Hz,1H),0.53(s,3H)( Figure 5 ). HRMS (ESI) m / z [MH] - calculated for C 41 H 55 Cl2N3O9Pt:998.2874; Found:998.2725( Figure 6 ).

[0120] Example 7 Synthesis of tetravalent platinum complex Pt4

[0121]

[0122] Referring to the synthesis of Pt1, cisPt(IV)-mNH2 (38.8 mg, 0.148 mmol) was replaced with carPt(IV)-mNH2 (45.8 mg, 0.148 mmol), and finally 93.6 mg of red solid powder was obtained with a yield of 69.6%.

[0123] 1 H NMR(400MHz, DMSO-d6)δ:8.73(s,1H),7.52(t,J=5.6Hz,1H),7.07(d,J=7.0Hz,1H),6.41(s,1H),6.34(d, J=7.2Hz,1H),6.12–5.58(m,6H),3.25–2.93(m,2H),2.59–2.49(m,2H),2.46–2.30(m,3H),2.25–2.15(m,3 H),2.09(s,3H),2.06–1.89(m,2H),1.85-1.64(m,3H),1.66–1.47(m,7H),1.43(d,J=13.6Hz,1H),1.37(s ,3H),1.29(d,J=18.3Hz,1H),1.21(s,3H),1.06(s,3H),1.01(s,3H),0.86(d,J=10.0Hz,1H),0.52(s,3H)( Figure 7 )。HRMS(ESI)m / z[M+H] + calculated for C 38 H 55 N3O 10 Pt:909.3611; Found:909.3604( Figure 8 ).

[0124] Example 8 Synthesis of tetravalent platinum complex Pt5

[0125]

[0126] Referring to the synthesis of Pt2, cisPt(IV)-diNH2 (35.2 mg, 0.074 mmol) was replaced with carPt(IV)-diNH2 (40.5 mg, 0.074 mmol), and finally 64.8 mg of red solid powder was obtained with a yield of 62.0%.

[0127] 1H NMR(400MHz,DMSO-d6)δ:8.66(s,2H),7.51(s,2H),7.03(s,2H),6.36(s,2H),6.29(s, 2H),6.19(s,6H),3.19–2.92(m,4H),2.44–2.28(m,6H),2.25–2.12(m,6H),2.05(s,6H) ),2.02–1.84(m,4H),1.80–1.65(m,4H),1.61–1.38(m,16H),1.33(s,6H),1.25(d,J=1 4.6Hz,2H),1.16(s,6H),1.02(s,6H),0.97(s,6H),0.82(d,J=9.2Hz,2H),0.47(s,6H)( Fig. 9 ). HRMS (ESI) m / z [MH] - calculated for C 70 H 96 N4O 14 Pt:1411.6509; Found:1410.6330( Fig.10 ).

[0128] Example 9 Synthesis of tetravalent platinum complex Pt6

[0129]

[0130] Referring to the synthesis of Pt3, Pt1 (35.2 mg, 0.074 mmol) was replaced with Pt4 (40.5 mg, 0.074 mmol), and finally 64.8 mg of reddish brown solid powder was obtained with a yield of 62.0%.

[0131] 1H NMR (400MHz, DMSO-d6) δ: 9.85 (s, 1H), 8.72 (s, 1H), 7.81 (d, J = 5.2Hz, 2H), 7.66 (t, J = 5.2Hz, 1H), 7.08 (t, J = 6.6 Hz,3H),6.40(s,1H),6.38(s,6H),6.34(d,J=7.0Hz,1H),4.72(s,2H),3.24–2.94(m,2H),2.47–2.34(m,5H),2.3 0–2.15(m,3H),2.08(s,3H),2.07–1.88(m,2H),1.87–1.67(m,3H),1.65–1.47(m,7H),1.42(d,J=14.6Hz,1H),1 .37(s,3H),1.28(d,J=12.6Hz,1H),1.20(s,3H),1.06(s,3H),1.01(s,3H),0.85(d,J=13.4Hz,1H),0.51(s,3H)( Fig.11 ). HRMS (ESI) m / z [MH] - calculated for C 47 H 61 N3O 13 Pt:1069.3772; Found:1069.3658( Fig.12 ).

[0132] Example 10 Synthesis of tetravalent platinum complex Pt7

[0133]

[0134] Referring to the synthesis of Pt1, cisPt(IV)-mNH2 (38.8 mg, 0.148 mmol) was replaced with oxaPt(IV)-mNH2 (48.2 mg, 0.148 mmol), and finally 86.8 mg of red solid powder was obtained with a yield of 62.7%.

[0135] 1H NMR(400MHz, DMSO-d6)δ:8.74(s,1H),8.34(s,1H),8.10(s,1H),7.80(s,1H),7.53(t,J=5.6Hz, 1H),7.17–7.03(m,2H),6.41(s,1H),6.35(d,J=7.2Hz,1H),3.13–2.96(m,2H),2.37(d,J=14.2H z,1H),2.30–2.15(m,3H),2.09(s,3H),2.06–1.72(m,7H),1.70–1.40(m,14H),1.38(s,3H),1.3 2(d,J=5.0Hz,1H),1.21(s,3H),1.06(s,3H),1.00(s,3H),0.86(d,J=13.0Hz,1H),0.52(s,3H)( Fig.13 ). HRMS (ESI) m / z [MH] - calculated for C 40 H 57 N3O 10 Pt:933.3611; Found:933.3535( Fig.14 ).

[0136] Example 11 Synthesis of tetravalent platinum complex Pt8

[0137]

[0138] Referring to the synthesis of Pt2, cisPt(IV)-diNH2 (35.2 mg, 0.074 mmol) was replaced with oxaPt(IV)-diNH2 (42.4 mg, 0.074 mmol), and finally 52.4 mg of red solid powder was obtained with a yield of 49.2%. 1H NMR(400MHz,DMSO-d6)δ:8.75(s,2H),8.29(s,2H),8.02(s,2H),7.63(s,2H),7.07(s,2H),6. 40(s,2H),6.34(d,J=6.8Hz,2H),3.18-3.06(m,4H),2.38(d,J=12.8Hz,2H),2.31-2.15(m,6H ),2.08(s,6H),2.05–1.89(m,5H),1.86–1.72(m,5H),1.68–1.40(m,22H),1.37(s,6H),1.29( d,J=3.6Hz,2H),1.20(s,6H),1.06(s,6H),1.00(s,6H),0.86(d,J=14.4Hz,2H),0.51(s,6H)( Fig.15 ). HRMS (ESI) m / z [MH] - calculated for C 72 H 98 N4O 14 Pt:1436.6648; Found:1436.6464( Fig.16 ).

[0139] Example 12 Synthesis of tetravalent platinum complex Pt9

[0140]

[0141] Referring to the synthesis of Pt3, Pt1 (35.2 mg, 0.074 mmol) was replaced with Pt7 (33.6 mg, 0.074 mmol), and finally 25.8 mg of reddish brown solid powder was obtained with a yield of 65.3%. 1H NMR (400MHz, DMSO-d6) δ: 9.86 (s, 1H), 8.72 (s, 1H), 8.37 (d, J = 26.8Hz, 2H), 8.13 (t, J = 7.4Hz, 1H), 7.92 (s, 1H), 7.82 (d ,J=8.6Hz,2H),7.74(t,J=5.0Hz,2H),7.06(d,J=8.4Hz,3H),6.40(s,1H),6.34(d,J=7.2Hz,1H),4.81(s,2H),3.09(m, 1H),2.40(d,J=14.4Hz,1H),2.33–2.25(m,2H),2.18(d,J=7.8Hz,1H),2.08(s,3H),2.07–1.73(m,7H),1.68–1.39(m,1 4H),1.37(s,3H),1.29(d,J=10.8Hz,1H),1.20(s,3H),1.06(s,3H),1.01(s,3H),0.85(d,J=13.2Hz,1H),0.51(s,3H)( Fig.17 ). HRMS (ESI) m / z [MH] - calculated for C 49 H 63 N3O 13 Pt:1095.3929; Found:1095.3796( Fig.18 ).

[0142] Example 13 In vitro antitumor activity test

[0143] The cells were cultured at 1×10 3 ~2×10 3 The cells were inoculated into 96-well culture plates at a density of 100 μg / well and incubated in a 37°C, 5% CO2 incubator for 24 h. Then, medium containing different concentrations of platinum drugs was added and incubated for 72 h. The cell activity was tested using the MTT method and the half inhibitory concentration (IC 50 Values) (Table 1). Among them, A549 is a human non-small cell lung cancer cell line, A549cisR is a drug-resistant strain of human non-small cell lung cancer cells, and MRC-5 is a human embryonic lung fibroblast cell line.

[0144] Table 1 In vitro cytotoxicity of tripterygium wilfordii-platinum (IV) complexes on cancer cells and normal cells [a]

[0145]

[0146]

[0147] Note: [a] The cells were incubated with the drug for 72 h, and the cell viability was measured by MTT assay and the IC of the complex was calculated. 50 [b]RF (resistance factor) is IC 50 (A549cisR) and IC 50 (A549) ratio. [c]SI (selectivity coefficient) is IC 50 (MRC-5) and IC 50 (A549). [d]FI (activity enhancement factor) is IC 50 (Pt(II) complex) and IC 50 (Pt(IV) complex) ratio.

[0148] The experimental results show that the in vitro cytotoxicity of the tetravalent platinum prodrugs Pt1-Pt9 involved in the present invention on cancer cell A549 and its drug-resistant strain A549cisR is superior to that of the corresponding divalent platinum complexes cisplatin (cDDP), carboplatin and oxaliplatin. In addition, the IC values ​​of Pt1-Pt9 on cisplatin-resistant A549cisR are 50 Value (3.97~17.63μmol·L -1 ) were lower than cisplatin (37.60 μmol·L -1 ), indicating that Pt1~Pt9 can overcome cisplatin resistance and effectively kill cisplatin-resistant cells.

[0149] Moreover, the in vitro cytotoxicity of the tetravalent platinum prodrugs Pt1 to Pt9 involved in the present invention on normal cells MRC-5 is lower than the toxicity of the corresponding divalent platinum complexes cisplatin (cDDP), carboplatin and oxaliplatin, and the mixture formed by directly mixing cisplatin (cDDP), carboplatin or oxaliplatin with celastrol, indicating that Pt1 to Pt9 have good cancer cell selectivity.

[0150] Example 14 Taking the tetravalent platinum compound Pt2 as an example, the situation of tetravalent platinum compound inducing cavitation of cancer cells

[0151] The cells were cultured at 2.5 × 10 5 The cells were inoculated into 6-well culture plates at a density of 10 cells / well and incubated in a 37°C, 5% CO2 incubator for 24 h before addition of 10 μmol·L -1 The culture medium containing tetravalent platinum prodrug Pt2 was incubated for 4 h, 8 h, and 12 h, and the morphology of the cells was observed under an inverted microscope and photographed for preservation.

[0152] Massive cytoplasmic vacuolization is the main feature of para-apoptosis. Fig.19 After incubation with Pt2, A549 cells and A549cisR cells showed large-scale cellular vacuolization. After 4 hours of incubation with Pt2, small vacuoles appeared in the cells, which gradually merged into large vacuoles (8 hours of treatment). After 12 hours, some cells became smaller and detached from the culture plate. The above changes in cell morphology are consistent with the morphological characteristics of para-apoptosis. This suggests that Pt2 may kill cancer cells by inducing para-apoptosis in cancer cells.

[0153] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. A tripterygium wilfordii-platinum (IV) complex, characterized in that: The general structure of the tripterygium wilfordii-platinum (IV) complex is shown in Formula S1: Among them, in formula S1, R represents One of them.

2. The method for preparing the tripterygium wilfordii-platinum (IV) complex according to claim 1, characterized in that: The steps include: (1) Cisplatin, carboplatin or oxaliplatin reacts with hydrogen peroxide at 50-75° C. to obtain a dihydroxy tetravalent platinum complex; the structural formula of the dihydroxy tetravalent platinum complex is as follows: (2) a dihydroxy tetravalent platinum complex is subjected to a condensation reaction with Fmoc-beta-alanine in the presence of a condensing agent and an organic base to obtain a tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc; the tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc has the following structural formula: (3) piperidine is added to the tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc obtained in step (2), respectively, and the reaction is carried out at room temperature with DMF as solvent; after the reaction, tetravalent platinum complexes cisPt(IV)-mNH2, carPt( IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2; the structural formula of the tetravalent platinum complex cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2 is as follows: (4) The tetravalent platinum complexes cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2 obtained in step (3) are subjected to condensation reaction with the ligand tripterygium wilfordii in the presence of a condensing agent and an organic base to obtain tetravalent platinum complexes Pt1, Pt2, Pt4, Pt5, Pt7 and Pt8 in sequence; the structures of the tetravalent platinum complexes Pt1, Pt2, Pt4, Pt5, Pt7 and Pt8 are as follows: (5) The tetravalent platinum complexes Pt1, Pt4, and Pt7 obtained in step (4) are condensed with 4-formylphenoxyacetic acid in the presence of a condensing agent and an organic base to obtain tetravalent platinum complexes Pt3, Pt6, and Pt9, respectively; the structures of the tetravalent platinum complexes Pt3, Pt6, and Pt9 are as follows:

3. The preparation method according to claim 2, characterized in that: In steps (2), (4) and (5), the condensing agent is any one of EDCi, DCC, DIC, HATU or HBTU, or a combination of any one of EDCi, DCC, DIC, HATU or HBTU and any one of NHS, HOBt or HOAt; and the organic base is any one of DMF, TEA or DIPEA.

4. The preparation method according to claim 2, characterized in that: In step (2), the reaction equivalent ratio of the dihydroxy tetravalent platinum complex to Fmoc-beta-alanine, the condensation agent, and the organic base is 0.2-1:1.2:1.4:0.

1.

5. The preparation method according to claim 2, characterized in that: In step (3), the reaction equivalent ratio of the tetravalent platinum complex cisPt(IV)-mFmoc, carPt(IV)-mFmoc, oxaPt(IV)-mFmoc, cisPt(IV)-diFmoc, carPt(IV)-diFmoc or oxaPt(IV)-diFmoc to piperidine is 1:13 to 26.

6. The preparation method according to claim 2, characterized in that: In step (4), the reaction equivalent ratio of the tetravalent platinum complex cisPt(IV)-mNH2, carPt(IV)-mNH2, oxaPt(IV)-mNH2, cisPt(IV)-diNH2, carPt(IV)-diNH2 or oxaPt(IV)-diNH2 with celastrol, condensation agent and organic base is 0.2-1:1.2:1.4:0.

1.

7. The preparation method according to claim 2, characterized in that: In step (5), the reaction equivalent ratio of the tetravalent platinum complex Pt1, Pt4 or Pt7 to 4-formylphenoxyacetic acid, condensation agent and organic base is 0.2-1:1.2:1.4:0.

1.

8. Use of the tripterygium wilfordii-platinum (IV) complex according to claim 1 in the preparation of anti-tumor drugs.

9. The use according to claim 8, characterized in that: The tripterygium wilfordii-platinum (IV) complex is one of the following structures:

10. The use according to claim 8, wherein the tumor is human non-small cell lung cancer cell A549 or human non-small cell lung cancer cell cisplatin-resistant strain A549cisR.

Images