Nitric oxide donor-type tetravalent platinum prodrug, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a nitric oxide (NO) donor-type tetravalent platinum prodrug of structural formula (I), a preparation method therefor, and a use thereof. The drug is selectively activated in a tumor cell, such that tetravalent platinum in the structure is reduced into cisplatin. The cisplatin acts as a bioorthogonal reaction catalyst to catalyze an NO donor fragment to release NO, and the latter and the cisplatin synergistically act to exert anti-tumor proliferation and anti-tumor metastasis activities. In contrast, the compound does not have the process in a normal cell, thereby achieving relatively high safety. In addition, the compound in the present invention has a unique carbamate long-carbon-chain structure. This structure can facilitate the binding of the prodrug to albumin, improving the cycling stability and pharmacokinetic characteristics of the drug, and achieving superior in-vivo anti-tumor proliferation and in-vivo anti-tumor metastasis activities.

Description

A nitric oxide donor tetravalent platinum prodrug and its preparation method and application Technical Field

[0001] The present invention relates to a medicine, a preparation method and an application thereof, and in particular to a nitric oxide donor type tetravalent platinum prodrug, a preparation method and an application thereof. Background Art

[0002] The incidence and mortality rates of malignant tumors are increasing worldwide. Chemotherapy is the standard treatment for cancer, and platinum (Pt) drugs are widely used for various tumors, accounting for approximately half of the available treatment options for cancer patients. Platinum derivatives, including cisplatin (DDP) and carboplatin, have garnered attention in the treatment of various cancers. However, despite their continued use, platinum drugs have numerous drawbacks, such as poor targeting.

[0003] Bioorthogonal chemistry

[0004] Bioorthogonal chemical reactions have the following characteristics: 1) reliability, selectivity, and orthogonality to other functions; 2) modularity and wide applicability; and 3) high yield. The most widely used bioorthogonal metal catalysts currently include gold, copper, and palladium. Since platinum and palladium are in the same main group of the periodic table, studies have shown that cisplatin can be used as a bioorthogonal catalyst to catalyze bond cleavage reactions. However, due to its strong cytotoxicity, cisplatin cannot be administered directly, so prodrug modification of cisplatin is required. The tetravalent platinum prodrug can be selectively reduced to cisplatin under the action of tumor cell reducing mediators. Cisplatin cross-links with DNA in tumor cells and also plays a bioorthogonal chemical catalytic role, releasing NO.

[0005] Quadrivalent platinum prodrugs

[0006] Tetravalent platinum prodrugs are a new class of molecules that may improve the pharmacological properties of divalent platinum anti-tumor drugs. 6 Compared with planar 5d 8 Platinum(II) complexes are kinetically more inert, a difference that allows tetravalent platinum compounds to be delivered as prodrugs with fewer side effects before reaching the target tumor site. Compared to Pt(II), the introduction of axial substituents in Pt(IV) can improve the molecule's pharmacokinetics, bioactivity, and targeting ability by modulating its reduction potential and lipophilicity. When tetravalent platinum complexes are reduced, two electrons are transferred to divalent platinum, which has antitumor activity, accompanied by the release of two axial ligands.

[0007] NO-donating drugs

[0008] NO donor drugs generally refer to prodrugs formed by NO donors and related drugs or certain active compounds through various linking groups. A variety of structural types of NO donors have been discovered, such as nitrosothiols, nitrates, NO-metal complexes (nitroprusside), furazan N-oxides, azodiolates, etc. Among them, azodiolates have obvious advantages in the selective and targeted release of NO. On the one hand, azodiolates are extremely unstable under physiological conditions and can automatically release 1 to 2 molecules of NO, with a half-life ranging from a few seconds to several hours. On the other hand, the O of azodiolates 2 The O connected to the nitrogen onium ion is called O 1 , the O connected to the olefinic nitrogen atom is O 2 ) after alkylation, it can be converted into a prodrug that is stable under physiological conditions; through certain specific enzyme recognition in the body or under the action of special physiological environment, O 2 Protective groups are converted into unstable azodicarbonium diolate anions, thereby achieving selective and targeted release of NO. 2 Protect new strategies.

[0009] Patent 202011077271.9 synthesizes an integrated prodrug based on bioorthogonal chemistry, which contains an organic compound of a tetravalent platinum complex and an azodicarbonium dialkoxide fragment. The compound is based on a tetravalent platinum complex and a nitric oxide donor molecule. Through a chemical coupling method, the nitric oxide donor molecule and the tetravalent platinum complex are connected by a succinate bond. As a whole molecule, it has anti-tumor activity. However, its circulation stability and pharmacokinetic properties need to be improved.

[0010] Objectives of the invention: The first objective of the present invention is to provide a nitric oxide donor-type tetravalent platinum prodrug with improved circulation stability and pharmacokinetic properties; the second objective of the present invention is to provide a method for preparing the nitric oxide donor-type tetravalent platinum prodrug; the third objective of the present invention is the application of the nitric oxide donor-type tetravalent platinum prodrug.

[0011] The nitric oxide donor tetravalent platinum prodrug of the present invention has the structural formula:

[0012] Wherein, R1 is piperazinyl or N-methylethanolamine, is cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum, R2 is selected from C4~C 18 .

[0013] The nitric oxide donor-type tetravalent platinum prodrug comprises a carbamate carbon chain, a tetravalent platinum complex and an azodicarbonium dialkoxide fragment. Based on the carbamate carbon chain, the tetravalent platinum complex and the nitric oxide donor molecule, the NO donor molecule and the tetravalent platinum carbamate carbon chain complex are connected by a chemical coupling method using a succinyl group to form a whole molecule.

[0014] Preferably, the R2 is selected from C6~C 12 .

[0015] Preferably, R2 is dodecyl.

[0016] Preferably, the It is cis-diaminedichlorodihydroxyplatinum.

[0017] Preferably, the R1 is piperazinyl.

[0018] Preferably, the structural formula of the nitric oxide donor type tetravalent platinum prodrug is:

[0019] The method for preparing the nitric oxide donor-type tetravalent platinum prodrug, when R1 is a piperazine group, comprises the following steps:

[0020] (1) Compound I4 undergoes an amide condensation reaction with succinic anhydride to obtain compound I5;

[0021] (2) Compound I5 undergoes esterification reaction with N-hydroxysuccinimide to obtain compound I6;

[0022] (3) Compound I6 undergoes an ester exchange reaction with cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum to obtain Compound I7;

[0023] (4) Compound I7 reacts with R2-N=C=O to produce compound I8;

[0024] The synthetic route is as follows:

[0025] Wherein, R2 is selected from C4~C 18 .

[0026] Synthesis of Compound I5: Dissolve Compound I4 in dichloromethane, add triethylamine as an acid-binding agent, and stir at 20-25°C. Then, add succinic anhydride. Stir and react at 20-25°C for 12-18 hours. Concentrate the reaction solution and perform column chromatography to obtain a yellow solid product I5. The molar ratio of Compound I4 to succinic anhydride is 1:1-1.5. The column chromatography stationary phase is silica gel, and the mobile phase is dichloromethane and methanol.

[0027] Synthesis of Compound I6: Compound I5 was placed in a single-necked reaction flask, and N-hydroxysuccinimide (NHS) and dimethyl carbonate (DCC) were added. Dichloromethane was then added for dissolution. The reaction mixture was stirred at 20-25°C for 2-4 hours. The filtrate was collected by filtration, concentrated to remove dichloromethane, and column chromatography was performed to obtain the desired product I6. The molar ratio of Compound I5 to NHS was 1:1-1.5. The column chromatography stationary phase was silica gel, and the mobile phase was dichloromethane and methanol.

[0028] Synthesis of Compound I7: Compound I6 was added to a single-necked reaction flask, along with cis- or trans-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum, and DMSO. The reaction mixture was stirred at 75-80°C in the dark for 4-5 hours. The yellow DMSO solution of I7 was then filtered and used directly in the next reaction. The molar ratio of Compound I6 to cis- or trans-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum was 1:1-1.5.

[0029] Synthesis of Compound Ⅰ8: Tetraalkyl isocyanate, octaalkyl isocyanate, dodecyl isocyanate, or octadecyl isocyanate was added dropwise to the yellow DMSO solution of Ⅰ7 and stirred at 20-25°C in the dark for 4-5 hours. After the reaction, saturated sodium chloride aqueous solution was added to the reaction solution, extracted with dichloromethane, and the organic layer was concentrated and column chromatography was performed to obtain the target product Ⅰ. 8a-e The stationary phase of the column chromatography is silica gel, and the mobile phase is dichloromethane and methanol. The molar ratio of the compound I7 to the alkyl isocyanate is 1:1 to 1.5.

[0030] The synthetic route of I4 is as follows:

[0031] Synthesis of Compound I2: A methanol solution of N-boc-piperazine and sodium methoxide was mixed and added to a polytetrafluoroethylene container. Tetrahydrofuran and anhydrous ether were then added. After the reaction system was displaced with nitrogen, nitric oxide (NO) gas was introduced to a pressure of 0.4-0.8 MPa. The reaction was carried out in a sealed container at 20-25°C for 40-48 hours. After the reaction was completed, the unreacted excess NO gas was released, and the pressure was reduced to atmospheric pressure. The container was opened and the reaction solution was poured into anhydrous ether to precipitate a large amount of white solid. The solid was filtered, and the filter cake was washed with ether and dried. The white product was collected as Compound I2.

[0032] Synthesis of Compound I3: Compound I2, pentadecacrown pentaether, and DMF were added to a 100 mL two-necked glass reaction flask. The flask was placed in an ice bath under N2 protection. Propylene bromide was then slowly added dropwise to the flask. The reaction was continued in an ice bath, and the reaction solution was then moved to 20°C-25°C for further reaction. After the reaction, DMF was first evaporated, and the residue was subjected to column chromatography to obtain a yellow solid, I3. The column chromatography stationary phase was silica gel, and the mobile phase was dichloromethane and methanol.

[0033] Synthesis of Compound I4: Dissolve Compound I3 in dichloromethane, stir at 20°C-25°C, and add saturated sodium bicarbonate solution until the pH is 7.5-8.0. After the reaction, wash with saturated sodium chloride solution, collect the organic layer, dry it, and concentrate it to obtain Compound I4.

[0034] The method for preparing the nitric oxide donor type tetravalent platinum prodrug, when R1 is N-methylethanolamine, comprises the following steps:

[0035] (1) Compound II3 undergoes an amide condensation reaction with succinic anhydride to obtain compound II4;

[0036] (2) Compound II4 undergoes esterification reaction with N-hydroxysuccinimide to obtain compound II5;

[0037] (3) Compound II5 undergoes an ester exchange reaction with cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum to obtain compound II6;

[0038] (4) Compound II6 undergoes an amination reaction with R2-N=C=O to obtain compound II7;

[0039] The synthetic route is as follows:

[0040] Wherein, R2 is selected from C4~C 18 .

[0041] Synthesis of Compound II4: Dissolve Compound II3 in anhydrous tetrahydrofuran, add 4-dimethylaminopyridine (DMAP), and stir at 20°C-25°C for 1-2 hours. Then, add succinic anhydride, and reflux the reaction mixture for 10-12 hours. For post-processing, filter the reaction mixture, concentrate the filtrate, add water, extract with dichloromethane, and concentrate the organic layer to obtain Compound II4. The molar ratio of Compound II3 to succinic anhydride is 1:1-1.5.

[0042] Synthesis of Compound II5: Compound II4 was placed in a single-necked reaction flask, and NHS and DCC were added. Dichloromethane was then added for dissolution. The reaction mixture was stirred at 20°C to 25°C for 2-4 hours. The filtrate was then filtered and concentrated to remove the dichloromethane. The residue was sanded and subjected to column chromatography to obtain the desired product II5. The column chromatography stationary phase was silica gel, and the mobile phases were dichloromethane and methanol. The molar ratio of Compound II4 to NHS was 1:1-1.5.

[0043] Synthesis of Compound II6: Compound II5 was added to a single-necked reaction flask, along with cis- or trans-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum and DMSO. The reaction mixture was stirred at 75-80°C in the dark for 4-5 hours. After post-treatment, the yellow DMSO solution of II6 was filtered and used directly in the next reaction. The molar ratio of Compound II5 to cis- or trans-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum was 1:1-1.5.

[0044] Synthesis of Compound II7: Tetraalkyl isocyanate, octaalkyl isocyanate, dodecyl isocyanate, or octadecyl isocyanate was dropwise added to a yellow DMSO solution of II6. The mixture was stirred at 20-25°C in the dark for 4-5 hours. Saturated aqueous sodium chloride solution was then added to the reaction mixture, and the mixture was extracted with dichloromethane. The dichloromethane was removed by concentration, and the target product II7 was obtained by column chromatography. The column chromatography was performed using silica gel as the stationary phase and dichloromethane and methanol as the mobile phase.

[0045] The synthetic route of compound II 3 is as follows:

[0046] Synthesis of Compound II2: N-methyl-2-hydroxyethylamine and a methanol solution of sodium methoxide were mixed and added to a polytetrafluoroethylene container. Anhydrous ether was added, and after displacing the reaction system with nitrogen, nitric oxide (NO) gas was introduced to a pressure of 0.4-0.8 MPa. The reaction was carried out in a sealed container at room temperature. After post-treatment, the unreacted excess NO gas was released and the pressure was reduced to atmospheric pressure. After opening the container, the reaction solution was poured into anhydrous ether to precipitate a large amount of white solid. The filter cake was filtered, washed with ether, and dried in a vacuum drying oven at room temperature. The white product was collected as Compound II2. The resulting Compound II2 was directly used in the next reaction without purification.

[0047] Synthesis of Compound II3: Compound II2 and DMF were added to a two-necked glass reaction flask. The flask was placed in an ice bath under N2 protection. Propylene bromide was then slowly added dropwise to the flask. The reaction was continued in an ice bath, and the reaction solution was then brought to room temperature for further reaction. Post-processing involved first removing the DMF by rotary evaporation, followed by column chromatography to obtain a colorless oil, II3. The column chromatography stationary phase was silica gel, and the mobile phase was dichloromethane and methanol.

[0048] The preparation method of the nitric oxide donor-type tetravalent platinum prodrug is described, wherein the compound I6 or II5 undergoes an ester exchange reaction with cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum, the solvent used is anhydrous DMSO, and the reaction conditions are dark, 75-80°C.

[0049] A pharmaceutical composition of the present invention contains the nitric oxide donor type tetravalent platinum prodrug and a pharmaceutically acceptable carrier.

[0050] Application of the nitric oxide donor type tetravalent platinum prodrug or its solvate in the prevention or treatment of anti-tumor drugs.

[0051] Mechanism of invention: The present invention integrates the two fragments of NO donor azodiol salt and tetravalent platinum complex, and adds a long alkyl chain part to the molecule to synthesize a new bioorthogonal autocatalytic NO donor / Pt(IV) prodrug. This drug can help the prodrug bind to serum albumin, so that the prodrug is bound to the surface of albumin, preventing it from being degraded by reducing substances in the circulation system, and improving the circulation stability and pharmacokinetic properties of the compounds involved in the aforementioned patent (ZL202011077271.9), thereby having better in vivo anti-tumor proliferation and in vivo anti-tumor metastasis activity. The prodrug can be selectively activated in tumor cells, and the tetravalent platinum in its structure is reduced to cisplatin. On the one hand, it cross-links with DNA in tumor cells, thereby exerting an anti-tumor effect; on the other hand, cisplatin acts as a bioorthogonal catalyst to catalyze O 2 The protected azodicarbonium diol salt undergoes a bioorthogonal bond cleavage reaction, resulting in the specific release of NO within tumor cells. The released NO can lead to the S-nitrosylation of the metal transporter antioxidant 1 copper chaperone (Atox1) and the copper ion transporting ATPase α peptide (P-type ATPase, ATP7a), resulting in reduced activity. This further inhibits the Cu loading of lysyl oxidase (LOX), increases the retention of Pt in cells, and exerts a synergistic anti-tumor effect. In normal cells, the compound does not undergo this process, thus having a better safety profile. Furthermore, the compounds of the present invention have a unique long carbon chain structure that helps the prodrug bind to albumin, improving the circulatory stability and pharmacokinetic properties of the tetravalent platinum prodrug, and further exhibiting excellent anti-tumor proliferation and anti-tumor metastasis activity in vivo.

[0052] Beneficial effects: Compared with the prior art, the present invention has the following advantages: The present invention synthesizes for the first time a new compound I with a unique carbamate carbon long chain structure 8a-e , II 7a and II 7b Surface plasmon resonance (SPR) biosensor experiments first demonstrated that 8c Binding ability to human serum albumin. This ability to bind to albumin makes the prodrug of the present invention have higher circulation stability and better pharmacokinetic properties than the compound in patent ZL202011077271.9, thereby making the compound contained in the present invention have better in vivo anti-tumor proliferation and anti-tumor metastasis activity. Anti-tumor mechanism studies have shown that compound I8c It preferentially enters tumor cells, where tetravalent platinum is reduced to cisplatin. At the same time, cisplatin catalyzes the compound to release NO, which leads to the S-nitrosylation of Atox1 and ATP7a, further inhibiting the Cu loading of lysyl oxidase LOX and increasing the retention of Pt in cells, thus achieving a synergistic anti-tumor effect of NO and platinum. Compared with the currently existing bioorthogonal prodrugs, compound I 8c It can avoid separate administration, enhance the targeting of the compound, and reduce the toxicity of the catalyst. Its long-chain carbamate carbon structure and the ability to bind to human serum albumin greatly improve the binding of compound I. 8c drugability. Description of the drawings:

[0053] FIG1A is a graph showing the determination of compound 11 (an analogue of the compound in ZL202011077271.9, not containing a long alkyl chain structure) by HPLC. 8a ,Ⅰ 8b ,Ⅰ 8c ,Ⅰ 8d , Ⅱ7 in rat plasma for 2 hours; Figure 1B ~ C is Ⅰ 8c The results of the determination of half-life in rat plasma are shown in FIG;

[0054] Figure 2 is compound Ⅰ 8a ,Ⅰ 8b ,Ⅰ 8c ,Ⅰ 8d , Ⅱ 7a , Cisplatin (DDP) at a concentration of 10 μM test results on the proliferation inhibition of different tumor cells;

[0055] Figure 3A~B is Ⅰ 8c The results of the SPR experiment on the binding constant KD of compound 11 and human serum albumin are shown in Figure 3C-D. 8c Computer docking results of binding with human serum albumin;

[0056] Figure 4A shows the same concentration of Ⅰ 8c After incubation of MCF-7 cells and MDA-MB-231 cells with cisplatin and cisplatin for 24 h, the Pt uptake of the cells was detected by ICP-MS; FIG4B is a graph showing the Pt uptake after incubation for 48 h using the same method;

[0057] Figure 5 shows the same concentration of Ⅰ 8c The results of intracellular NO release after incubation with MCF-7 cells, MCF-7 / DDP cells, and MDA-MB-231 cells;

[0058] FIG6A is a diagram of compound I 8cTranswell assay was performed to verify the ability of compound I to inhibit the migration of MDA-MB-231 cells; FIG6B is a graph showing the effect of compound I on the migration of MDA-MB-231 cells. 8c The results of a scratch test to verify its ability to inhibit the migration of MDA-MB-231 cells;

[0059] Figure 7 is Ⅰ 8c Figures show the experimental results of the efficacy evaluation of the drug against MDA-MB-231 xenograft tumors in mice; Figure 7A shows the results of tumor volume measurement and calculation every other day; Figure 7B shows the results of mouse body weight measurement every other day; Figure 7C shows the results of mouse tumor weighing; and Figure 7D shows an image of the mouse tumor.

[0060] Figure 8 is Ⅰ 8c Figure 8A shows the results of an in vivo anti-MDA-MB-231-luc lung metastasis experiment in mice; Figure 8A shows three representative small animal fluorescence imaging images; Figure 8B shows the statistical calculation of the fluorescence intensity in the lungs of mice in each group;

[0061] Figure 9A is a diagram of the biotin-switch method used to verify Ⅰ 8c The experimental results show that the released NO causes the increased nitrosylation of Atox1 and ATP7a in MDA-MB-231 cells; FIG9B is a grayscale statistical graph of the bands. DETAILED DESCRIPTION

[0062] The present invention will be further described below with reference to specific embodiments.

[0063] Example 1

[0064] The nitric oxide donor type tetravalent platinum prodrug of the present invention, Ⅰ 8a The preparation method comprises the following steps:

[0065] (1) Synthesis of Compound I2:

[0066] N-boc-piperazine (I1, 10 g) and 12.18 g of a methanol solution of sodium methoxide (5.4 mol / L) were mixed and added to a polytetrafluoroethylene container. 20 ml of tetrahydrofuran and 120 mL of anhydrous ether were added. After the reaction system was replaced with nitrogen, nitric oxide (NO) gas was introduced to a pressure of 0.4-0.8 MPa. The reaction was carried out in a sealed container at room temperature for 48 hours. After the reaction was completed, the unreacted excess NO gas was released to reduce the pressure to atmospheric pressure. The container was opened and the reaction solution was poured into 1 L of anhydrous ether to precipitate a large amount of white solid. The solid was filtered, and the filter cake was washed three times with ether. The solid was then dried in a vacuum oven for 2 hours. The white product was collected as compound I2.

[0067] (2) Synthesis of Compound I3:

[0068] 4 g of compound I2 (14.9 mmol), pentadecacrown pentaether (0.149 mmol), and 25 mL of DMF were added to a 100 mL two-necked glass reaction flask. The flask was placed in an ice bath under N2 protection. Then, 2.22 mL of propargyl bromide (29.8 mmol) was slowly added dropwise to the flask. The reaction was continued in an ice bath for 0.5 h, then brought to room temperature and continued for 12 h. After the reaction, DMF was first removed by rotary evaporation, and the residue was subjected to column chromatography to obtain a yellow solid I3. The column chromatography stationary phase was silica gel, and the mobile phase was a 20:1 ratio of dichloromethane to methanol.

[0069] (3) Synthesis of Compound I4:

[0070] Dissolve 1 g (3.52 mmol) of compound I3 in 10 mL of dichloromethane. Add 3 mL of dichloromethane and stir at room temperature for 2 h. Then add saturated sodium bicarbonate solution until the pH reaches 8.0. After the reaction, wash three times with saturated sodium chloride solution. Collect the organic layer, dry it, and concentrate it to obtain compound I4.

[0071] (4) Synthesis of Compound I5:

[0072] 1.1 g of compound Ⅰ4 (5.97 mmol) was dissolved in 40 mL of dichloromethane, and 1.2 g of triethylamine (11.9 mmol) was added. The mixture was stirred at room temperature for 15 minutes, followed by the addition of 1.2 g of succinic anhydride (11.94 mmol). The mixture was stirred overnight at room temperature. The reaction solution was concentrated and column chromatography was performed to obtain a yellow solid product Ⅰ5. The column chromatography stationary phase was silica gel, and the mobile phase was a 10:1 ratio of dichloromethane to methanol.

[0073] (5) Synthesis of Compound I6:

[0074] To a single-necked reaction flask, 100 mg of compound Ⅰ5 (0.175 mmol) was added, along with 21.9 mg of NHS (0.19 mmol) and 39.2 mg of DCC (0.19 mmol). Dissolved in 3 mL of dichloromethane, the reaction mixture was stirred at room temperature for 30 minutes. The filtrate was collected by filtration, concentrated to remove the dichloromethane, and then purified by column chromatography to yield the desired product Ⅰ6. The column chromatography stationary phase was silica gel, and the mobile phase was a 10:1 ratio of dichloromethane to methanol.

[0075] (6) Synthesis of Compound Ⅰ7

[0076] Place 86.5 mg of compound I6 (0.23 mmol) in a single-necked reaction flask, add 76.8 mg of cis-diaminedichlorodihydroxyplatinum (cisplatin), and add 2 mL of DMSO. Stir the reaction mixture at 80°C in the dark for 5 h. Filter to obtain a yellow DMSO solution of I7, which can be used directly in the next reaction.

[0077] (7) Compound Ⅰ 8a Synthesis

[0078] Tetraalkyl isocyanate (0.368 mmol) was added dropwise to the yellow DMSO solution of Ⅰ7 and stirred at room temperature in the dark for 5 h. After the reaction, saturated sodium chloride aqueous solution was added to the reaction solution, and the mixture was extracted three times with dichloromethane. The organic layer was concentrated and the target product Ⅰ was obtained by column chromatography. 8a .

[0079] Compound Ⅰ 8a : Pale yellow solid, 43 mg. 1 H NMR(300MHz,Chloroform-d)δ6.18(s,6H),5.73–5.51(m,1H),4.80(s,2H),3.72(d,J=22.5Hz,4H),3.60–3.40(m,4H),3.21– 2.89(m,2H),2.68(s,2H),2.60(d,J=11.6Hz,2H),2.24(s,1H),1.48–1.35(m,2H),1.33–1.26(m,2H),0.88(t,J=7.1Hz,3H). 13 C NMR(75MHz,DMSO-d6)δ180.03,170.39,163.86,79.00,78.46,60.58,50.63,50.38,43.20,31.95,30.83,28.49,19.54,13.76.HRMS(ESI)calculated for C 16 H 31 Cl2N7O7Pt,[M+H] + :699.13880; found:699.13859.ppm error 0.3.

[0080] Example 2

[0081] The nitric oxide donor type tetravalent platinum prodrug of the present invention, Ⅰ 8b The preparation method comprises the following steps:

[0082] Steps (1) to (7) are the same as in Example 1, except that octaalkyl isocyanate is used instead of tetraalkyl isocyanate in step (7).

[0083] Compound Ⅰ 8b : Pale yellow solid, 58 mg. 1 H NMR(300MHz,Chloroform-d)δ6.16(s,6H),5.58(s,1H),4.79(s,2H),3.74(s,2H),3.67(s,2H),3.53(s,2H),3.48(s,2H),3.04(s ,2H),2.79–2.63(m,2H),2.62–2.53(m,2H),2.25(d,J=35.0Hz,1H),1.49–1.35(m,2H),1.31–1.20(m,10H),0.86(t,J=6.5Hz,3H). 13 C NMR(75MHz,DMSO-d6)δ180.03,170.38,162.28,78.97,78.44,60.58,50.62,50.38,43.1 9,31.24,30.76,29.82,28.82,28.69,28.49,26.43,22.07,13.92.HRMS(ESI)calculated for C 20 H 39 Cl2N7O7Pt,[M+H] + :755.20140; found:755.19994.ppm error 1.9.

[0084] Example 3

[0085] The nitric oxide donor type tetravalent platinum prodrug of the present invention, Ⅰ 8c The preparation method comprises the following steps:

[0086] Steps (1) to (7) are the same as those in Example 1, except that dodecyl isocyanate is used instead of tetraalkyl isocyanate in step (7).

[0087] Compound Ⅰ 8c : Pale yellow solid, 69 mg. 1H NMR(300MHz,Chloroform-d)δ6.17(s,6H),5.58(s,1H),4.80(d,J=2.2Hz,2H),3.75(s,2H),3.67(s,2H),3.53(s,2H),3.48(s,2H ),3.03(s,2H),2.81–2.65(m,2H),2.61–2.55(m,2H),2.25(s,1H),1.54–1.34(m,2H),1.31–1.19(m,18H),0.86(t,J=6.5Hz,3H). 13 C NMR(75MHz,Chloroform-d)δ182.75,171.82,163.82,77.24,76.72,61.19,50.92,50.80,43 .95,31.92,30.03,29.73,29.68,29.54,29.37,27.10,22.68,14.10.HRMS(ESI)calculated for C 24 H 47 Cl2N7O7Pt,[M+H] + :811.26400; found:811.26082.ppm error 3.9.

[0088] Example 4

[0089] The nitric oxide donor type tetravalent platinum prodrug of the present invention, Ⅰ 8d The preparation method comprises the following steps:

[0090] Steps (1) to (7) are the same as in Example 1, except that octadecyl isocyanate is used instead of tetraalkyl isocyanate in step (7).

[0091] Compound Ⅰ 8d : Pale yellow solid, 63 mg. 1 H NMR(300MHz,Chloroform-d)δ6.15(s,6H),5.54(s,1H),4.80(d,J=2.5Hz,2H),3.75(s,2H),3.67(t,J=3.9Hz,2H),3.54(s,2H),3.49 (s,2H),3.04(s,2H),2.68(s,2H),2.60(t,J=2.3Hz,2H),2.17(s,1H),1.50–1.35(m,2H),1.30–1.21(m,30H),0.86(t,J=6.4Hz,3H). 13C NMR(75MHz,Chloroform-d)δ182.73,171.82,77.23,76.61,61.16,50.87,43.92,40.56 ,31.92,30.01,29.75,29.67,29.52,29.36,27.08,22.68,14.10.HRMS(ESI)calculated for C 30 H 59 Cl2N7O7Pt,[M+H] + :895.35590; found:895.35242.ppm error 3.8.

[0092] Example 5

[0093] The nitric oxide donor type tetravalent platinum prodrug of the present invention, Ⅰ 8e The preparation method comprises the following steps:

[0094] Steps (1) to (7) are the same as those in Example 1, except that in step (6), trans-diaminedichlorodihydroxyplatinum (trans-platinum) is used instead of cis-diaminedichlorodihydroxyplatinum; and in step (7), dodecyl isocyanate is used instead of tetraalkyl isocyanate.

[0095] Compound Ⅰ 8e : Pale yellow solid, 39 mg. 1 H NMR(300MHz,Chloroform-d)δ6.12(s,6H),5.68(s,1H),4.80(s,2H),4.26(t,J=5.0Hz,2H),3.64(t,J=4.9,4.0Hz,2 H),3.12–2.95(m,5H),2.65(s,3H),2.54(s,2H),1.49–1.34(m,2H),1.24–1.18(m,18H),0.84(t,J=6.5,5.6Hz,3H). 13 C NMR(75MHz,Chloroform-d)δ181.63,173.38,163.89,77.88,76.97,61.59,61.26,52.69,42.36,41.36 ,31.97,31.05,30.50,30.12,29.79,29.73,29.61,29.41,27.19,22.71,14.11.HRMS(ESI)calculated for C 23 H 46 Cl2N6O8Pt,[M+H] +:800.24802; found:800.24657.ppm error 1.8.

[0096] Example 6

[0097] The nitric oxide donor type tetravalent platinum prodrug of the present invention, II 7a The preparation method comprises the following steps:

[0098] (1) Synthesis of compound II2:

[0099] N-methyl-2-hydroxyethylamine (II1, 20 mL) and 50.79 mL of a methanol solution of sodium methoxide (5.4 mol / L) were mixed and added to a polytetrafluoroethylene container. 200 mL of anhydrous ether was added. After replacing the reaction system with nitrogen, nitric oxide (NO) gas was introduced to a pressure of 0.4-0.8 MPa. The reaction was carried out in a sealed container at room temperature for 24 hours. After post-treatment, the unreacted excess NO gas was released and the pressure was reduced to atmospheric pressure. After opening the container, the reaction solution was poured into 1 L of anhydrous ether to precipitate a large amount of white solid. The filter cake was washed three times with ether and dried in a vacuum drying oven at room temperature for 2 hours. The white product was collected as Compound II2. The obtained Compound II2 was directly used in the next reaction without purification.

[0100] (2) Synthesis of compound II3:

[0101] 1 g of compound II2 and 5 mL of DMF were added to a 100 mL two-necked glass reaction flask. The flask was placed in an ice bath under N2 protection. 757.8 mg of propargyl bromide was then slowly added dropwise to the flask. The reaction was continued in an ice bath for 0.5 h, then brought to room temperature and continued for 12 h. Post-processing began by removing the DMF by rotary evaporation. Column chromatography afforded compound II3 as a colorless oil. The column chromatography stationary phase was silica gel, and the mobile phase was a 20:1 ratio of dichloromethane to methanol.

[0102] (3) Synthesis of compound II4:

[0103] Dissolve 700 mg of compound II3 in 10 mL of anhydrous tetrahydrofuran, add 122 mg of DMAP, and stir at room temperature for 15 minutes. Then, add 608 mg of succinic anhydride. Reflux the reaction mixture overnight. For post-treatment, filter the reaction mixture, concentrate the filtrate, add water, and extract five times with dichloromethane. Concentrate the organic layer to obtain compound II4.

[0104] (4) Synthesis of Compound II5:

[0105] 48 mg of compound II4 (0.175 mmol) was placed in a single-necked reaction flask, followed by the addition of 21.9 mg of NHS (0.19 mmol) and 39.2 mg of DCC (0.19 mmol). Dissolved in 3 mL of dichloromethane, the reaction mixture was stirred at room temperature for 30 minutes. The filtrate was then filtered and concentrated to remove the dichloromethane. The residue was sanded and subjected to column chromatography to yield the desired product II5. The column chromatography stationary phase was silica gel, and the mobile phase was a 50:1 ratio of dichloromethane to methanol.

[0106] (5) Synthesis of Compound II6:

[0107] 85 mg of compound II5 was placed in a single-necked reaction flask, along with 76.8 mg of cis-diaminedichlorodihydroxyplatinum and 2 mL of DMSO. The reaction mixture was stirred at 80°C in the dark for 5 h. After post-treatment, the yellow DMSO solution of II6 was filtered and used directly in the next reaction.

[0108] (6) Compound II 7a Synthesis of:

[0109] To a yellow DMSO solution of II6 (0.184 mmol) was added dodecyl isocyanate (0.368 mmol) dropwise and stirred at room temperature in the dark for 5 h. Saturated sodium chloride aqueous solution was then added to the reaction solution, extracted three times with dichloromethane, concentrated to remove dichloromethane, and column chromatography was performed to obtain the target product II. 7a The stationary phase of the column chromatography was silica gel, and the mobile phase was dichloromethane and methanol in a volume ratio of 50:1.

[0110] Compound II 7a : Pale yellow solid, 38 mg. 1 H NMR(300MHz,Chloroform-d)δ6.12(s,6H),5.68(s,1H),4.80(s,2H),4.26(t,J=5.0Hz,2H),3.64(t,J=4.9,4.0Hz,2 H),3.12–2.95(m,5H),2.65(s,3H),2.54(s,2H),1.49–1.34(m,2H),1.24–1.18(m,18H),0.84(t,J=6.5,5.6Hz,3H). 13C NMR(75MHz,Chloroform-d)δ181.63,173.38,163.89,77.88,76.97,61.59,61.26,52.69,4 2.36,41.36,31.97,31.05,30.50,30.12,29.79,29.73,29.61,29.41,27.19,22.71,14.11.

[0111] Example 7

[0112] The nitric oxide donor type tetravalent platinum prodrug of the present invention, II 7b The preparation method comprises the following steps:

[0113] Steps (1) to (6) are the same as those in Example 5, except that in step (5), trans-diaminedichlorodihydroxyplatinum is used instead of cis-diaminedichlorodihydroxyplatinum; and in step (7), dodecyl isocyanate is used instead of tetraalkyl isocyanate.

[0114] Compound II 7b : Pale yellow solid, 38 mg. 1 H NMR(500MHz,Chloroform-d)δ6.82,6.72,6.45,4.52,4.25,3.79,3.76,3.72,3.69,3.10, 3.01,2.86,2.70,2.64,1.47,1.34,1.28,1.27,1.25,1.25,1.25,1.23,1.22,1.22,0.89.

[0115] Performance Testing

[0116] (1) Drug stability test in rat plasma

[0117] Compound 11 (an analogue of the compound in ZL202011077271.9, not containing a long alkyl chain structure), Ⅰ 8a ,Ⅰ 8b ,Ⅰ 8c ,Ⅰ 8d , Ⅱ 7a Stability in rat plasma, and Ⅰ 8c Half-life determination in rat plasma.

[0118] HPLC (Innovai ODS-2 column 5 μm, The stability of the compound in rat plasma was determined by incubating the compound (0.5 mM) in rat plasma at 37°C and recording the HPLC spectrum after 2 hours. 8c, spectra were recorded at 0, 12, 24, 48, and 72 hours to determine the half-life.

[0119] The test results are shown in Figure 1. Figure 1A is a HPLC determination of compound 11 (an analogue of the compound in ZL202011077271.9, not containing a long alkyl chain structure), I 8a ,Ⅰ 8b ,Ⅰ 8c ,Ⅰ 8d , Ⅱ7 in rat plasma for 2 hours, compounds with long alkyl chains showed significant improvement in stability. Figure 1B-C is Ⅰ 8c The results of the determination of half-life in rat plasma show that Ⅰ 8c The half-life is approximately 30 hours.

[0120] (2) Test of drug proliferation inhibition activity on different tumor cells

[0121] Determination of compound Ⅰ 8a ,Ⅰ 8b ,Ⅰ 8c ,Ⅰ 8d , Ⅱ7, the proliferation inhibitory activity of cisplatin (DDP) on different tumor cells at a concentration of 10μM.

[0122] MTT assay for compound I 8a ,Ⅰ 8b ,Ⅰ 8c ,Ⅰ 8d , Ⅱ 7a The inhibitory activity of cisplatin (DDP) at a concentration of 10 μM on the proliferation of different tumor cells, including triple-negative breast cancer cells MDA-MB-231, MDA-MB-468, breast cancer cells MCF-7, MCF7 / DDP, non-small cell lung cancer cells A549, A549 / DDP, ovarian cancer cells A2780, and colon cancer cells HCT116.

[0123] The test results are shown in Figure 2. All compounds showed certain anti-tumor activity against different tumor cells. 8b ,Ⅰ 8c ,Ⅰ 8d The activities of Ⅱ and Ⅱ7 were significantly better than those of cisplatin.

[0124] According to the above preliminary screening results, select Ⅰ 8c IC values ​​of MDA-MB-231, MCF-7, A549 / DDP, MCF-7 / DDP, and MCF-10A cells were further determined. 50 values, and cisplatin (DDP) and 11 were selected as controls.

[0125] The test results are shown in Table 1. In the tested cells, compound I 8c The activity was significantly better than that of cisplatin and 11, indicating that Ⅰ 8c It has better anti-tumor activity.

[0126] Table 1.IC 50 (μM) of 2, 10c and DDP against MDA-MB-231, MCF-7, A549 / DDP, MCF-7 / DDP and MCF-10A cells. a a Cells were treated with the indicated compounds for 72h, and the cell viability and IC 50 values ​​were determined by MTT assay.Data were expressed as the mean±SD from three individual experiments. b FI values, fold increase, calculated as IC 50 (DDP) / ID 50 (I 8c ). c ND: not determined.

[0127] (3) Drug binding test to human serum albumin

[0128] SPR assay I 8c The binding constant (KD) of compound 11 to human serum albumin, and Ⅰ 8c In silico docking results of binding to human serum albumin.

[0129] A Biacore T200 instrument (GE Healthcare) was used with PBS-P running buffer (10 mM phosphate buffer containing 2.7 mM KCl, 137 mM NaCl, and 0.05% surfactant P20) at 25°C. Human serum albumin was immobilized on a sensor CM5 chip using a standard amine coupling procedure (10 mM sodium acetate, pH 5.5). The compounds under investigation were serially diluted and then passed over the sensor chip at a flow rate of 30 μL / min for 120 seconds during the contact phase, followed by a 120-second injection of buffer during the dissociation phase. KD values ​​were calculated using Biacore T200 Evaluation Software, version 1.0 (GE Healthcare).

[0130] The test results are shown in Figure 3A. 8c The KD value for human serum albumin was 15.35 μM. In sharp contrast, compound 11 (Figure 3B), an analog of the compound in patent ZL202011077271.9, which does not contain a long alkyl chain moiety, had a KD value that exceeded the detection limit and could not be determined, which emphasizes the key role of the long alkyl chain in human serum albumin binding affinity.

[0131] (4)Ⅰ 8c , Pt uptake of DDP in MCF-7 cells and MDA-MB-231 cells.

[0132] The same concentration (1 μmol / L) of Ⅰ 8c MCF-7 cells and MDA-MB-231 cells were incubated with cisplatin for 24 h (as shown in FIG4A ) and 48 h (as shown in FIG4B ), and then the Pt uptake of different cells was observed by ICP-MS. The test results are shown in FIG4 .

[0133] As shown in Figure 4, compound I 8c The uptake of Pt is much higher than that of cisplatin. Part of the reason is that compound I 8c The lipid solubility and stability of Ⅰ are much higher than those of cisplatin. 8c The released NO causes the nitrosation of metal transporters Atox1 and ATP7a and reduces their activity, thereby reducing the efflux of platinum by metal transporters in tumor cells.

[0134] (5) Compound Ⅰ 8c Intracellular NO release.

[0135] Detection of compound Ⅰ using DAF-FM DA fluorescent probe 8c The test results of NO release in different cells (flow cytometry) are shown in FIG5 .

[0136] From Figure 5, we can see that Ⅰ 8c The NO release in tumor cells MDA-MB-231 and MCF-7 / DDP was greater than that in normal mammary epithelial cells MCF-10A, indicating that compound I 8c It selectively degrades and catalyzes the release of NO in tumor cells and has good biocompatibility with normal cells.

[0137] (6) Compound Ⅰ 8c Effects on the migration ability of MDA-MB-231 cells

[0138] Next, we studied compound I 8c To prevent the antiproliferative activity of the compound from having a significant impact on the migration ability of MDA-MB-231 cells, first, the IC values ​​of the compound on tumor cells were calculated according to the above method. 50 The calculation method of the value (MTT method) was used to first calculate the IC value of the compound. 10 The value is 84.477nM.

[0139] Secondly, the transwell migration assay demonstrated that compound I 8c It can significantly inhibit the migration ability of MDA-MB-231 cells. The transwell migration assay measures the migration ability of cells by counting the number of migrated cells. The greater the number of cells, the stronger the migration ability. As shown in Figure 6A, the experimental results show that compound I 8c It can significantly inhibit the migration ability of MDA-MB-231 cells.

[0140] Then we continued to use 85nM I 8c The scratch test was performed, and the results are shown in Figure 6B. 8c The degree of scratch closure of the group was lower than that of the control group, indicating that Ⅰ 8c Significantly inhibited the migration ability of MDA-MB-231 cells.

[0141] (7) Compound Ⅰ 8c Metabolic properties in the body

[0142] To further evaluate compound I 8c The metabolic properties of compound I were determined in vivo. 8c The pharmacokinetic (PK) properties of the original drug and total platinum in rats were studied. Three male SD rats were selected and intravenously injected with Ⅰ 8c Blood was collected from the fundus venous plexus before administration and at 5 minutes, 15 minutes, 30 minutes, 60 minutes, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after administration (5 mg / kg). The upper plasma layer was centrifuged and stored at -20°C until further use. The test results are shown in Table 2.

[0143] Table 2.PK Parameters of I 8c (iv 5mg / kg). a a Values ​​are the average of three determinations.

[0144] From Table 2, we can see that Ⅰ 8c The half-life in vivo is 0.29h, T max is 0.08h, and its AUC (0-t) The total platinum half-life in rats is 23.58h, T max is 0.08h, and its AUC (0-t) It is 14720.28ug / L*h.

[0145] (8) Compound Ⅰ 8c Acute toxicity

[0146] Compound I 8c Acute toxicity test, observe compound Ⅰ 8c Acute toxicity is to determine the safe range of drug use. A preliminary experiment was first conducted, which showed that the test drug had certain toxicity. Intravenous injection of 70 mg / kg of the drug caused the death of 4 / 4 mice, while intravenous injection of 30 mg / kg caused the death of 0 / 4 mice. Based on the experimental results of the preliminary experiment, we selected the following doses of 63 mg / ml, 56.7 mg / ml, 51.03 mg / ml, 45.93 mg / ml, and 41.34 mg / ml for LD50. 50 Experiment: The drug was administered intravenously once at the above-mentioned dose, and the poisoning symptoms and death of mice in each group were recorded. The dead animals were autopsied.

[0147] The experimental results are as follows:

[0148] Abnormal reaction: After intravenous injection of the drug into mice, the animals in each dose group became listless, paralyzed and eventually died. The higher the dose, the more pronounced the symptoms and the shorter the time to death. The results of this experiment showed that compound I 8c It is toxic to mice to a certain extent and can cause death in mice after administration at higher doses. 8c LD after intravenous administration 50 The value is 53.2507 (48.3854~58.6053) mg / kg.

[0149] (9) Compound Ⅰ 8c Evaluation of drug efficacy in MDA-MB-231 mouse transplanted tumors

[0150] The efficacy of MDA-MB-231 cell transplanted tumors in mice was tested. MDA-MB-231 cells were inoculated into the second pair of mammary pads on the right side of the mice and the tumors were established and grew to 100 mm. 3 The mice were randomly divided into groups. 8c treatment group, a negative control group that only received the vehicle, and a DDP group. 8c (5, 2.5 and 1.25 mg / kg, intravenous injection, once every three days), and the DDP group was 5 mg / kg, intravenous injection, once every three days.

[0151] The results are shown in Figure 7. Figure 7A shows the results of measuring and calculating the tumor volume every other day; Figure 7B shows the results of measuring the mouse body weight every other day; Figure 7C shows the results of weighing the mouse tumor; and Figure 7D shows a picture of the mouse tumor. 8c The growth of tumor cells was inhibited in a dose-dependent manner, with Ⅰ 8c The tumor inhibition rate of the 5mg / kg group was 71.08%, while that of the 5mg / kg cisplatin group was only 58.51%. 8c The in vivo pharmacodynamic activity of Ⅰ is significantly better than that of cisplatin. 8c The effects of high, medium and low dose groups on the body weight of mice were lower than those of the cisplatin group, indicating that compound I 8c It has a better safety profile than cisplatin.

[0152] (10) Compound Ⅰ 8c In vivo anti-MDA-MB-231-luc lung metastasis

[0153] Pair I 8c The anti-MDA-MB-231 cell lung metastasis activity was tested in mice. Six-week-old female BALB / c mice were injected with MDA-MB-231 / luc cells (5×10 7 cells). One week later, the animals were randomly divided into three groups, with six mice in each group. 8c The treatment group received 2.5 mg / kg of DDP via the tail vein every three days. The DDP-treated group also received 2.5 mg / kg of DDP via the tail vein every three days. The negative control group received only the vehicle at the same frequency. For imaging, mice were anesthetized with 2% inhaled isoflurane and intraperitoneally injected with D-luciferin. Signals were acquired 10 minutes later using an IVIS Lumina imaging system. The test results are shown in Figure 8; Figure 8A shows three representative small animal fluorescence imaging images; Figure 8B shows a statistical graph of the quantitative fluorescence intensity in the lungs of mice in each group.

[0154] As shown in Figure 8, compared with the control group and DDP group, 8cThe treated mice showed significantly lower fluorescence intensity on day 12, indicating that Ⅰ 8c Treatment significantly inhibited the lung metastasis of MDA-MB-231 in vivo.

[0155] (11)Ⅰ 8c Test for changes in Atox1 and ATP7a nitrosation levels in tumor cells

[0156] Compound I 8c After incubation with MDA-MB-231 cells, the nitrosation levels of Atox1 and ATP7a were detected using the Biotin-Switch method.

[0157] The test results are shown in Figure 9, where Figure 9A is the result of using the Biotin-Switch method to verify the 8c The experimental results show that the released NO causes the increase of Atox1 and ATP7a nitrosylation in MDA-MB-231 cells; Figure 9B is the grayscale statistical graph of the bands. 8c The released NO significantly increased the S-nitrosation of Atox1 and ATP7a. Increased nitrosation of these two transporters could lead to reduced metal binding activity and enhanced retention of platinum in cancer cells, thereby enhancing the integration of prodrug I 8c It has more effective anti-tumor effect.

Claims

1. A nitric oxide-donating tetravalent platinum prodrug, characterized in that, Its structural formula is: Among them, R1 is a piperazinyl group or an N-methylethanolamine group, It is cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum, and R2 is selected from C4-C 18 .

2. The nitric oxide-donating tetravalent platinum prodrug according to claim 1, wherein The R2 is selected from C6-C 12 .

3. The nitric oxide-donating tetravalent platinum prodrug according to claim 1, wherein R2 is dodecyl.

4. The nitric oxide-donating tetravalent platinum prodrug according to claim 1, wherein The said is cis-diamine-dichloro-dihydroxy platinum.

5. The nitric oxide-donating tetravalent platinum prodrug according to claim 1, wherein R1 is piperazinyl.

6. A method for preparing a nitric oxide donor type tetravalent platinum prodrug according to claim 1, characterized in that, When R1 is piperazinyl, it includes the following steps: (1) Compound I4 undergoes an amide condensation reaction with succinic anhydride to obtain compound I5; (2) Compound I5 undergoes an esterification reaction with N-hydroxysuccinimide to obtain compound I6; (3) Compound I6 undergoes a transesterification reaction with cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum to obtain compound I7; (4) Compound I7 undergoes an amination reaction with R2-N=C=O to obtain compound I8; The synthetic route is as follows: Among them, R2 is selected from C4-C 18 .

7. A method for preparing the nitric oxide-donating tetravalent platinum prodrug according to claim 1, characterized in that, When R1 is N-methylethanolamine group, it includes the following steps: (1) Compound II3 undergoes an amide condensation reaction with succinic anhydride to obtain compound II4; (2) Compound II4 undergoes an esterification reaction with N-hydroxysuccinimide to obtain compound II5; (3) Compound II5 undergoes a transesterification reaction with cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum to obtain compound II6; (4) Compound II6 undergoes an amination reaction with R2-N=C=O to obtain compound II7; The synthetic route is as follows: Among them, R2 is selected from C4-C 18 .

8. The preparation method of the nitric oxide-donating tetravalent platinum prodrug according to claim 6 or 7, characterized in that, In the transesterification reaction of the compound I6 or II5 with cis-diaminedichlorodihydroxyplatinum or trans-diaminedichlorodihydroxyplatinum, the solvent used is anhydrous DMSO, the reaction condition is light avoidance, and the reaction temperature is 75-80 °C.

9. A pharmaceutical composition, characterized in that, Containing the nitric oxide-donating tetravalent platinum prodrug according to any one of claims 1-5 and a pharmaceutically acceptable carrier.

10. Use of the nitric oxide-donating tetravalent platinum prodrug according to any one of claims 1-5 or its solvate in the prevention or treatment of anti-tumor drugs.

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