A nitric oxide donor type tetravalent platinum prodrug, and a preparation method and application thereof

Abstract

The application discloses a kind of structural formula as I nitric oxide donor type tetravalent platinum prodrug and its preparation method and application, the drug is selectively activated in tumor cell, and the tetravalent platinum in its structure is reduced to cisplatin, while cisplatin is used as biological orthogonal reaction catalyst, catalyzes NO donor fragment to release NO, the latter cooperates with cisplatin and plays antitumor proliferation and anti-tumor metastasis activity;And in normal cell, compound does not have the above process, to have good security;And the compound in the application has unique carbamate carbon long chain structure, this structure can help prodrug and albumin are combined, improve the circulation stability and pharmacokinetic characteristics of drug, and have better in vivo antitumor proliferation and in vivo anti-tumor metastasis activity.

Description

Technical Field

[0001] This invention relates to a drug, its preparation method and application, and particularly to a nitric oxide donor-type tetravalent platinum prodrug, its preparation method and application. Background Technology

[0002] Currently, the incidence and mortality rates of malignant tumors are rising globally. Chemotherapy is the standard treatment for cancer, with platinum (Pt) drugs widely used for various tumors, accounting for about half of the available treatment options for cancer patients. Platinum derivatives, including cisplatin (DDP) and carboplatin, are gaining attention in the treatment of various cancers. However, despite their continued use, platinum drugs still have many drawbacks, such as poor targeting.

[0003] Bioorthogonal Chemistry

[0004] Bioorthogonal chemistry possesses several advantages, including: 1) reliability, selectivity, and orthogonality with other functional groups; 2) modularity and wide applicability; and 3) high yield. The most widely used metal catalysts in bioorthogonal chemistry currently include gold, copper, and palladium. Since platinum and palladium are in the same group of the periodic table, studies have shown that cisplatin can serve as a bioorthogonal catalyst for bond-breaking reactions. However, due to its strong cytotoxicity, cisplatin cannot be directly administered, thus requiring prodrug modification. Tetravalent platinum prodrugs can be selectively reduced to cisplatin under the action of tumor cell reducing mediators. Cisplatin then cross-links with DNA within tumor cells, simultaneously exerting a bioorthogonal catalytic effect to release NO.

[0005] Quadrivalent platinum predrug

[0006] Tetravalent platinum prodrugs are a new class of molecules that may improve the pharmacological properties of divalent platinum antitumor drugs. They possess octahedral low-spin 5d... 6 Platinum(IV) complexes compared to 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 into Pt(IV) can improve the molecule's pharmacokinetics, bioactivity, and targeting ability by modulating reduction potential and lipophilicity. When tetravalent platinum complexes are reduced, two electrons are transferred, reducing them to divalent platinum with antitumor activity, accompanied by the release of two axial ligands.

[0007] NO donor type drugs

[0008] NO donor drugs generally refer to prodrugs formed by linking NO donors and related drugs or certain active compounds through various connecting groups. Various structural types of NO donors have been discovered, such as nitrosothiols, nitrate esters, NO-metal complexes (nitroprussides), furazolidone N-oxides, and azodium diolates; among them, azodium diolates have significant advantages in the selective and targeted release of NO. On the one hand, azodium diolates are extremely unstable under physiological conditions, spontaneously releasing 1-2 molecules of NO, with half-lives ranging from seconds to hours. On the other hand, the O2 of azodium diolates... 2 The O atom bonded to the nitrogen ion is called the O atom bond. 1 The O atom bonded to the nitrogen atom in the alkene bond is O. 2 After alkylation, it can be converted into a prodrug that is stable under physiological conditions; through recognition by certain specific enzymes in vivo or under the action of special physiological environments, the O2 is removed. 2 The protecting group transforms into an unstable azotumene glycol anion, thereby achieving selective and targeted NO release. To date, various O2-based ... 2 New protection strategies.

[0009] Patent 202011077271.9 describes the synthesis of an integrative prodrug based on bioorthogonal chemistry, comprising an organic compound containing a tetravalent platinum complex and an azomonium glycol salt fragment. This compound is based on a tetravalent platinum complex and a nitric oxide donor molecule, linked by a succinate bond via chemical coupling, forming a single molecule with antitumor activity. However, its cyclic stability and pharmacokinetic properties require further improvement. Summary of the Invention

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

[0011] The nitric oxide donor-type tetravalent platinum prodrug of this invention has the following structural formula:

[0012]

[0013] Wherein, R1 is piperazine or N-methylethanolamine. R2 is either cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum, and R2 is selected from C4 to C5. 18 .

[0014] The nitric oxide donor-type tetravalent platinum prodrug comprises a carbamate carbon chain, a tetravalent platinum complex, and an azomonium glycol salt 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 linked together by a chemical coupling method using a succinyl group to form a single molecule.

[0015] Preferably, R2 is selected from C6 to C6. 12 .

[0016] Preferably, R2 is dodecyl.

[0017] Preferably, the It is cis-diamine dichlorodihydroxyplatinum.

[0018] Preferably, R1 is piperazine-based.

[0019] Preferably, the structural formula of the nitric oxide donor-type tetravalent platinum prodrug is:

[0020]

[0021] The method for preparing the nitric oxide donor-type tetravalent platinum prodrug, when R1 is piperazine-based, includes the following steps:

[0022] (1) Compound I4 undergoes an amide condensation reaction with succinic anhydride to give compound I5;

[0023] (2) Compound I5 undergoes an esterification reaction with N-hydroxysuccinimide to give compound I6;

[0024] (3) Compound I6 undergoes an ester exchange reaction with cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum to give compound I7;

[0025] (4) Compound I7 undergoes an amination reaction with R2-N=C=O to give compound I8;

[0026] The synthesis route is as follows:

[0027]

[0028] Among them, R2 is selected from C4 to C6. 18 .

[0029] Synthesis of Compound I5: Compound I4 was dissolved in dichloromethane, and triethylamine was added as an acid-binding agent. The mixture was stirred at 20–25°C, and then succinic anhydride was added. The reaction mixture was stirred at 20–25°C for 12–18 h. After concentration, column chromatography was used to obtain a yellow solid product I5. The molar ratio of compound I4 to succinic anhydride was 1:1–1.5. The stationary phase for column chromatography was silica gel, and the mobile phase was dichloromethane and methanol.

[0030] Synthesis of Compound I6: Compound I5 was placed in a single-necked reaction flask, and N-hydroxysuccinimide (NHS), dimethyl carbonate (DCC), and dichloromethane were added to dissolve it. The reaction mixture was stirred at 20–25°C for 2–4 h. The filtrate was collected by filtration, concentrated to remove dichloromethane, and column chromatography was used to obtain the target product I6. The molar ratio of compound I5 to NHS was 1:1–1.5; the stationary phase of the column chromatography was silica gel, and the mobile phase was dichloromethane and methanol.

[0031] Synthesis of Compound I7: Compound I6 was placed in a single-necked reaction flask, and either cis- or trans-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum was added. DMSO was then added, and the reaction mixture was stirred at 75–80°C in the dark for 4–5 hours. The resulting yellow DMSO solution of I7 was then filtered and can be used directly in the next reaction step. The molar ratio of Compound I6 to cis- or trans-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum was 1:1–1.5.

[0032] Synthesis of compound I8: Tetraalkyl isocyanate, octaalkyl isocyanate, dodecyl isocyanate, or octadecyl isocyanate was added dropwise to a yellow DMSO solution of I7, and the mixture was stirred at 20–25°C in the dark for 4–5 h. After the reaction was complete, saturated sodium chloride aqueous solution was added to the reaction mixture, and the solution was extracted with dichloromethane. The organic layer was concentrated, and column chromatography was used 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 compound I7 to alkyl isocyanate is 1:1 to 1.5.

[0033] The synthetic route for I4 is as follows:

[0034]

[0035] 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 diethyl ether were added, and the reaction system was purged with N2. Nitric oxide (NO) gas was then introduced to bring the pressure to 0.4–0.8 MPa. The reaction was carried out at 20–25°C under sealed conditions for 40–48 h. After the reaction was completed, the excess unreacted NO gas was released, and the pressure was reduced to atmospheric pressure. The container was then opened, and the reaction solution was poured into anhydrous diethyl ether, precipitating a large amount of white solid. The solid was filtered, the filter cake was washed with diethyl ether, dried, and the white product was collected as Compound I2.

[0036] Synthesis of Compound I3: Compound I2, pentadecanoate, and DMF were added to a 100 mL two-necked glass reaction flask. The flask was placed in an ice bath under N2 protection. Bromopropyne was then slowly added dropwise to the flask. After the addition was complete, the reaction continued in an ice bath. The reaction mixture was then transferred to 20–25 °C to continue the reaction. After the reaction was complete, DMF was first evaporated, and the residue was purified by column chromatography to obtain a yellow solid, I3. The stationary phase of the column chromatography was silica gel, and the mobile phase was dichloromethane and methanol.

[0037] Synthesis of compound I4: Compound I3 was dissolved in dichloromethane and stirred at 20°C–25°C. Saturated sodium bicarbonate solution was added until the pH reached 7.5–8.0. After the reaction was complete, the mixture was washed with saturated sodium chloride solution, the organic layer was collected, dried, and concentrated to obtain compound I4.

[0038] The method for preparing the nitric oxide donor-type tetravalent platinum prodrug, when R1 is N-methylethanolamine, includes the following steps:

[0039] (1) Compound II3 undergoes an amide condensation reaction with succinic anhydride to give compound II4;

[0040] (2) Compound II4 undergoes an esterification reaction with N-hydroxysuccinimide to give compound II5;

[0041] (3) Compound II5 undergoes an ester exchange reaction with cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum to give compound II6;

[0042] (4) Compound II6 undergoes an amination reaction with R2-N=C=O to give compound II7;

[0043] The synthesis route is as follows:

[0044]

[0045] Among them, R2 is selected from C4 to C6. 18 .

[0046] Synthesis of Compound II4: Compound II3 was dissolved in anhydrous tetrahydrofuran, and 4-dimethylaminopyridine (DMAP) was added. The mixture was stirred at 20℃–25℃ for 1–2 h, followed by the addition of succinic anhydride. The reaction mixture was refluxed for 10–12 h. Post-treatment: The reaction mixture was first filtered, the filtrate was concentrated, water was added, and the mixture was extracted with dichloromethane. The organic layer was concentrated to obtain Compound II4. The molar ratio of Compound II3 to succinic anhydride was 1:1–1.5.

[0047] Synthesis of Compound II5: Compound II4 was placed in a single-necked reaction flask, NHS and DCC were added, and dichloromethane was added to dissolve it. The reaction solution was stirred at 20℃~25℃ for 2~4h. Then, the filtrate was collected by filtration, concentrated to remove dichloromethane, and the residue was prepared into sand. Column chromatography was used to obtain the target product II5. The stationary phase of the column chromatography was silica gel, and the mobile phase was dichloromethane and methanol. The molar ratio of compound II4 to NHS was 1:1~1.5.

[0048] Synthesis of Compound II6: Compound II5 was placed in a single-necked reaction flask, and cis- or trans-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum and DMSO were added. The reaction solution was stirred at 75–80 °C in the dark for 4–5 h. After post-treatment, a yellow DMSO solution of II6 was obtained by filtration and used directly in the next reaction step. The molar ratio of compound II5 to cis- or trans-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum was 1:1–1.5.

[0049] Synthesis of compound II7: Tetraalkyl isocyanate, octaalkyl isocyanate, dodecyl isocyanate, or octadecyl isocyanate was added dropwise to a yellow DMSO solution of II6, and the mixture was stirred at 20–25°C in the dark for 4–5 h. Subsequently, a saturated aqueous sodium chloride solution was added to the reaction mixture, and the solution was extracted with dichloromethane. The dichloromethane was then concentrated to remove it, and column chromatography was used to obtain the target product II7. The stationary phase of the column chromatography was silica gel, and the mobile phase was dichloromethane and methanol.

[0050] The synthetic route for compound II3 is as follows:

[0051]

[0052] Synthesis of Compound II2: A mixture of N-methyl-2-hydroxyethylamine and a methanol solution of sodium methoxide was added to a polytetrafluoroethylene (PTFE) container. Anhydrous diethyl ether was added, and the reaction system was purged with N2. Nitric oxide (NO) gas was then introduced to achieve a pressure of 0.4–0.8 MPa, and the reaction was carried out in a sealed environment at room temperature. Afterward, 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 diethyl ether, precipitating a large amount of white solid. This solid was filtered, the filter cake was washed with diethyl ether, and dried in a vacuum oven at room temperature. The white product collected was compound II2. Compound II2 was used directly in the next reaction without purification.

[0053] 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. Bromopropyne was then slowly added dropwise to the flask. After the addition was complete, the reaction continued in an ice bath. The reaction mixture was then cooled to room temperature to continue the reaction. Post-treatment: DMF was first removed by rotary evaporation, followed by column chromatography to obtain a colorless oily substance, II3. The stationary phase for the column chromatography was silica gel, and the mobile phase was dichloromethane and methanol.

[0054] The method for preparing the nitric oxide donor type tetravalent platinum prodrug, wherein in the transesterification reaction of compound I6 or II5 with cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum, the solvent used is anhydrous DMSO, and the reaction conditions are protected from light and 75-80°C.

[0055] A pharmaceutical composition of the present invention comprises the nitric oxide donor-type tetravalent platinum prodrug and a pharmaceutically acceptable carrier.

[0056] The use of the nitric oxide donor-type tetravalent platinum prodrug or its solvate in the prevention or treatment of antitumor drugs.

[0057] Mechanism of Invention: This invention integrates two fragments—azoium glycol salt of NO donor and a tetravalent platinum complex—and adds a long alkyl chain moiety to the molecule, synthesizing a novel bioorthogonal autocatalytic NO donor / Pt(IV) prodrug. This drug facilitates the binding of the prodrug to serum albumin, allowing it to remain beneath the albumin surface and preventing degradation by reducing substances in the circulatory system. This improves the cyclic stability and pharmacokinetic properties of the compounds involved in the aforementioned patent (ZL202011077271.9), thereby exhibiting superior in vivo antitumor proliferation and antitumor metastasis activity. This prodrug can be selectively activated in tumor cells, where the tetravalent platinum in its structure is reduced to cisplatin. On one hand, cisplatin cross-links with DNA within tumor cells, thereby exerting an antitumor effect; on the other hand, cisplatin acts as a bioorthogonal catalyst, catalyzing the oxidation of NO within tumor cells. 2 The protected azomonium glycol salt undergoes a bioorthogonal bond cleavage reaction, resulting in the specific release of NO within tumor cells. This released NO leads to S-nitrosylation of the metal transporter Antioxidant 1 copper chaperone (Atox1) and the P-type ATPase α-peptide (ATP7a), reducing their activity. This further inhibits the Cu load of lysyl oxidase (LOX), increasing Pt retention in cells and exerting a synergistic antitumor effect. The compound does not exhibit this process in normal cells, thus demonstrating better safety. Furthermore, the compound in this invention possesses a unique long carbon chain structure, which facilitates the binding of the prodrug to albumin, improving the cyclic stability and pharmacokinetic properties of the tetravalent platinum prodrug, and further generating excellent antitumor proliferation and antitumor metastasis activity in vivo.

[0058] Beneficial effects: Compared with the prior art, the present invention has the following advantages: The present invention is the first to synthesize a new compound I with a unique carbamate carbon long chain structure. 8a-e II 7a and II 7bThe surface plasmon resonance (SPR) biosensor was first used to demonstrate that I... 8c The binding affinity to human serum albumin. This albumin-binding affinity gives the prodrug of this invention higher cyclic stability and better pharmacokinetic properties compared to the compound in patent ZL202011077271.9, thereby giving the compound contained in this invention superior in vivo antitumor proliferation and antitumor metastasis activity. Antitumor mechanism studies have shown that compound I... 8c It preferentially enters tumor cells, where tetravalent platinum is reduced to cisplatin. Simultaneously, cisplatin catalyzes the release of NO, which leads to S-nitrosylation of Atox1 and ATP7a, further inhibiting the Cu load of lysyl oxidase LOX and increasing Pt retention in cells, thus exerting a synergistic antitumor effect of NO and platinum. Compared with currently existing bioorthogonal prodrugs, compound I... 8c This avoids separate administration, enhances the compound's targeting, and reduces catalyst toxicity. Its long carbamate carbon chain structure significantly improves the binding affinity of compound I to human serum albumin. 8c Its medicinal properties. Attached image description:

[0059] Figure 1 A is determined by HPLC using compounds 11 (analogs of compounds in ZL202011077271.9, excluding alkyl long-chain structures), and I. 8a Ⅰ 8b Ⅰ 8c Ⅰ 8d Stability data of II7 in rat plasma for 2 hours; Figure 1 B to C are I 8c The results of half-life determination in rat plasma are shown in the figure.

[0060] Figure 2 It is compound I 8a Ⅰ 8b Ⅰ 8c Ⅰ 8d II 7a Figure 1 shows the results of the assay for the inhibitory activity of cisplatin (DDP) at a concentration of 10 μM on the proliferation of different tumor cells.

[0061] Figure 3 A to B are I 8c Results of the assay for the binding constant KD of compound 11 and human serum albumin (SPR). Figure 3 CD is I 8c A computer-generated image showing the binding of serum albumin to human serum albumin.

[0062] Figure 4A is the same concentration of I 8c After incubating MCF-7 cells and MDA-MB-231 cells with cisplatin for 24 h, respectively, the Pt uptake of the cells was detected by ICP-MS. Figure 4 B is a graph showing Pt uptake after 48 hours of incubation using the same method.

[0063] Figure 5 It is the same concentration of I 8c Figure showing the intracellular NO release results after incubation with MCF-7 cells, MCF-7 / DDP cells, and MDA-MB-231 cells;

[0064] Figure 6 A is compound I. 8c Figure showing the experimental results of Transwell assays to verify its ability to inhibit the migration of MDA-MB-231 cells; Figure 6 B is compound I. 8c Figure showing the experimental results of scratch assay to verify its ability to inhibit the migration of MDA-MB-231 cells;

[0065] Figure 7 It is I 8c Figure 1 shows the experimental results evaluating the efficacy of anti-MDA-MB-231 xenograft drugs in mice; among them, Figure 7 A represents the results of tumor volume measurement and calculation every other day; Figure 7 B represents the mouse's weight measured every other day; Figure 7 C represents the tumor weight measurement result in mice; Figure 7 D is an image of a mouse tumor;

[0066] Figure 8 It is I 8c Figure 1 shows the experimental results of anti-MDA-MB-231-luc lung metastasis in mice; among them, Figure 8 A consists of three representative fluorescence imaging images of small animals; Figure 8 B is a statistical graph showing the quantitative calculation of fluorescence intensity in the lungs of mice in each group;

[0067] Figure 9 A is to verify I using the Biotin-Switch method. 8c The experimental results showing that the released NO caused an increase in Atox1 and ATP7a nitrosation in MDA-MB-231 cells; Figure 9 B is its grayscale statistical chart. Detailed Implementation

[0068] The present invention will be further described below with reference to specific embodiments.

[0069] Example 1

[0070] The nitric oxide donor-type tetravalent platinum prodrug of this invention, I 8a The preparation method includes the following steps:

[0071] (1) Synthesis of compound I2:

[0072]

[0073] 10 g of N-boc piperazine (I1) and 12.18 g of a 5.4 mol / L sodium methoxide methanol solution were mixed and added to a polytetrafluoroethylene container. 20 mL of tetrahydrofuran and 120 mL of anhydrous diethyl ether were added, and the reaction system was purged with N2. Nitric oxide (NO) gas was then introduced to bring the pressure to 0.4–0.8 MPa, and the reaction was carried out at room temperature under a sealed environment for 48 h. After the reaction was complete, the excess unreacted NO gas was released, and the pressure was reduced to atmospheric pressure. The container was opened, and the reaction solution was poured into 1 L of anhydrous diethyl ether, precipitating a large amount of white solid. The solid was filtered, and the filter cake was washed three times with diethyl ether. The mixture was then dried in a vacuum drying oven for 2 h, and the white product collected was compound I2.

[0074] (2) Synthesis of compound I3:

[0075]

[0076] 4 g of compound I2 (14.9 mmol), pentadecanoate (0.149 mmol), and 25 mL of DMF were added to a 100 mL two-necked glass flask. The flask was placed in an ice bath under N2 protection. Then, 2.22 mL of bromopropyne (29.8 mmol) was slowly added dropwise to the flask. After the addition was complete, the reaction was continued in an ice bath for 0.5 h, and then the reaction mixture was moved to room temperature and the reaction was continued for 12 h. After the reaction was completed, DMF was first removed by rotary evaporation, and the residue was purified by column chromatography to obtain a yellow solid I3. The stationary phase of the column chromatography was silica gel, and the mobile phase was dichloromethane to methanol in a volume ratio of 20:1.

[0077] (3) Synthesis of compound I4:

[0078]

[0079] 1 g (3.52 mmol) of compound I3 was dissolved in 10 mL of dichloromethane. 3 mL of dichloromethane was added, and the mixture was stirred at room temperature for 2 h. Saturated sodium bicarbonate solution was then added until the pH reached 8.0. After the reaction was complete, the mixture was washed three times with saturated sodium chloride solution. The organic layer was collected, dried, and concentrated to obtain compound I4.

[0080] (4) Synthesis of compound I5:

[0081]

[0082] 1.1 g of compound I4 (5.97 mmol) was dissolved in 40 mL of dichloromethane. 1.2 g of triethylamine (11.9 mmol) was added, and the mixture was stirred at room temperature for 15 min. Then, 1.2 g of succinic anhydride (11.94 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated, and column chromatography was used to obtain a yellow solid product I5. The stationary phase for column chromatography was silica gel, and the mobile phase was dichloromethane to methanol in a volume ratio of 10:1.

[0083] (5) Synthesis of compound I6:

[0084]

[0085] 100 mg of compound I5 (0.175 mmol) was placed in a single-necked reaction flask, and 21.9 mg of NHS (0.19 mmol) and 39.2 mg of DCC (0.19 mmol) were added. 3 mL of dichloromethane was added to dissolve the compound, and the reaction mixture was stirred at room temperature for 30 mins. The filtrate was collected by filtration, concentrated to remove dichloromethane, and column chromatography was performed to obtain the target product I6. The stationary phase for column chromatography was silica gel, and the mobile phase was dichloromethane to methanol in a volume ratio of 10:1.

[0086] (6) Synthesis of compound I7

[0087]

[0088] 86.5 mg (0.23 mmol) of compound I6 was placed in a single-necked reaction flask, and 76.8 mg of cis-diamine dichlorodihydroxyplatinum (cisplatin) was added. 2 mL of DMSO was then added, and the reaction mixture was stirred at 80 °C in the dark for 5 h. The solution was then filtered to obtain a yellow DMSO solution of I7, which can be used directly in the next reaction step.

[0089] (7) Compound I 8a Synthesis

[0090]

[0091] Tetraalkyl isocyanate (0.368 mmol) was added dropwise to a yellow DMSO solution of I7, and the mixture was stirred at room temperature in the dark for 5 h. After the reaction was complete, saturated sodium chloride aqueous solution was added to the reaction mixture, and the solution was extracted three times with dichloromethane. The organic layer was concentrated and column chromatography was used to obtain the target product I. 8a .

[0092] Compound I 8a Pale yellow solid, 43 mg. 1H 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 forC 16 H 31 Cl2N7O7Pt,[M+H] + :699.13880; found:699.13859.ppm error 0.3.

[0093] Example 2

[0094] The nitric oxide donor-type tetravalent platinum prodrug of this invention, I 8b The preparation method includes the following steps:

[0095]

[0096] Steps (1) to (7) are the same as in Example 1, except that octaalkyl isocyanate is used instead of tetraalkyl isocyanate in step (7).

[0097] 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). 13C 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.

[0098] Example 3

[0099] The nitric oxide donor-type tetravalent platinum prodrug of this invention, I 8c The preparation method includes the following steps:

[0100]

[0101] Steps (1) to (7) are the same as in Example 1, except that in step (7), dodecyl isocyanate is used instead of tetraalkyl isocyanate.

[0102] Compound I 8c Pale yellow solid, 69mg. 1 H 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.

[0103] Example 4

[0104] The nitric oxide donor-type tetravalent platinum prodrug of this invention, I 8d The preparation method includes the following steps:

[0105]

[0106] Steps (1) to (7) are the same as in Example 1, except that octadecyl isocyanate is used instead of tetraalkyl isocyanate in step (7).

[0107] Compound I 8d Pale yellow solid, 63mg. 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). 13 C 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.

[0108] Example 5

[0109] The nitric oxide donor-type tetravalent platinum prodrug of this invention, I 8e The preparation method includes the following steps:

[0110]

[0111] Steps (1) to (7) are the same as in Example 1, except that in step (6), trans-diamine dichlorodihydroxyplatinum (transplatinum) is used instead of cis-diamine dichlorodihydroxyplatinum; and in step (7), dodecyl isocyanate is used instead of tetraalkyl isocyanate.

[0112] Compound I 8e Pale yellow solid, 39mg. 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.

[0113] Example 6

[0114] The nitric oxide donor-type tetravalent platinum prodrug of this invention, II 7a The preparation method includes the following steps:

[0115] (1) Synthesis of compound II2:

[0116]

[0117] N-methyl-2-hydroxyethylamine (II1, 20 mL) and 50.79 mL of a 5.4 mol / L sodium methoxide methanol solution were mixed and added to a polytetrafluoroethylene container. 200 mL of anhydrous diethyl ether was added, and the reaction system was purged with N2. Nitric oxide (NO) gas was then introduced to bring the pressure to 0.4-0.8 MPa, and the reaction was carried out in a sealed environment for 24 hours. Afterwards, 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 1 L of anhydrous diethyl ether, precipitating a large amount of white solid. The solid was filtered, and the filter cake was washed three times with diethyl ether. The cake was then dried in a vacuum drying oven at room temperature for 2 hours. The white product collected was compound II2. Compound II2 was used directly in the next reaction without purification.

[0118] (2) Synthesis of compound II3:

[0119]

[0120] 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. Then, 757.8 mg of bromopropyne was slowly added dropwise to the reaction flask. After the addition was complete, the reaction was continued in an ice bath for 0.5 h. The reaction solution was then transferred to room temperature and the reaction was continued for 12 h. Post-treatment involved removing DMF by rotary evaporation, followed by column chromatography to obtain a colorless oily substance, II3. The stationary phase for column chromatography was silica gel, and the mobile phase was dichloromethane to methanol in a volume ratio of 20:1.

[0121] (3) Synthesis of compound II4:

[0122]

[0123] 700 mg of compound II3 was dissolved in 10 mL of anhydrous tetrahydrofuran, and 122 mg of DMAP was added. After stirring at room temperature for 15 mins, 608 mg of succinic anhydride was added, and the reaction solution was refluxed overnight. For post-treatment, the reaction solution was first filtered, the filtrate was concentrated, water was added, and the mixture was extracted five times with dichloromethane. The organic layer was concentrated to obtain compound II4.

[0124] (4) Synthesis of compound II5:

[0125]

[0126] 48 mg of compound II4 (0.175 mmol) was placed in a single-necked reaction flask, and 21.9 mg of NHS (0.19 mmol) and 39.2 mg of DCC (0.19 mmol) were added. 3 mL of dichloromethane was added to dissolve the compound, and the reaction mixture was stirred at room temperature for 30 mins. The filtrate was then collected by filtration, concentrated to remove dichloromethane, and the residue was prepared as a precipitate. Column chromatography was used to obtain the target product II5. The stationary phase for column chromatography was silica gel, and the mobile phase was dichloromethane to methanol in a volume ratio of 50:1.

[0127] (5) Synthesis of compound II6:

[0128]

[0129] 85 mg of compound II5 was placed in a single-necked reaction flask, and 76.8 mg of cis-diamine dichlorodihydroxyplatinum and 2 mL of DMSO were added. The reaction solution was stirred at 80 °C in the dark for 5 h. After post-treatment, the solution was filtered to obtain a yellow DMSO solution of II6, which was used directly in the next step of the reaction.

[0130] (6) Compound II 7a Synthesis:

[0131]

[0132] Dodecyl isocyanate (0.368 mmol) was added dropwise to a yellow DMSO solution of II6 (0.184 mmol), and the mixture was stirred at room temperature in the dark for 5 h. Then, a saturated sodium chloride aqueous solution was added to the reaction mixture, and the solution was extracted three times with dichloromethane. The dichloromethane was then concentrated to remove the dichloromethane, and column chromatography was used to obtain the target product II. 7a The stationary phase for column chromatography was silica gel, and the mobile phase was dichloromethane to methanol in a product ratio of 50:1.

[0133] 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.

[0134] Example 7

[0135] The nitric oxide donor-type tetravalent platinum prodrug of this invention, II 7b The preparation method includes the following steps:

[0136]

[0137] Steps (1) to (6) are the same as in Example 5, except that in step (5), trans-diamine dichlorodihydroxyplatinum is used instead of cis-diamine dichlorodihydroxyplatinum; and in step (7), dodecyl isocyanate is used instead of tetraalkyl isocyanate.

[0138] 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.

[0139] Performance testing

[0140] (1) Stability test of the drug in rat plasma

[0141] Compound 11 (an analogue of compound ZL202011077271.9, excluding alkyl long-chain structures), I 8a Ⅰ 8b Ⅰ 8c Ⅰ 8d II 7a Stability in rat plasma, and I 8c Determination of half-life in rat plasma.

[0142] Using HPLC (Innovai ODS-2 column 5μm), The stability of the compound in rat plasma was determined using an HPLC microscope (1250 × 4.60 mm). The compound (0.5 mM) was incubated in rat plasma at 37 °C, and the HPLC chromatogram was recorded after 2 hours. For I... 8c Spectra were recorded at 0, 12, 24, 48, and 72 hours to determine the half-life.

[0143] Test results are as follows Figure 1 As shown, Figure 1 A is determined by HPLC using compounds 11 (analogs of compounds in ZL202011077271.9, excluding alkyl long-chain structures) and I. 8a I 8b I 8c I 8d Stability data of II7 in rat plasma for 2 hours showed that compounds with long alkyl chains exhibited significantly improved stability. Figure 1 BC is I 8c The results of the half-life determination in rat plasma are shown in the figure. 8c The half-life is approximately 30 hours.

[0144] (2) Test of the inhibitory activity of the drug on the proliferation of different tumor cells

[0145] Determination of compound I 8a I 8b I 8c I 8d The inhibitory activity of cisplatin (DDP) on the proliferation of different tumor cells at a concentration of 10 μM.

[0146] Determination of compound I using the MTT assay 8a I 8b I 8c I 8d II 7a The inhibitory activity of cisplatin (DDP) at a concentration of 10 μM against the proliferation of different tumor cells, including triple-negative breast cancer cells MDA-MB-23l, 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.

[0147] Test results are as follows Figure 2 As shown, all compounds exhibited certain antitumor activity against different tumor cells. Compound I... 8b I 8c I 8d Both II7 and β2-II7 have significantly better activity than cisplatin.

[0148] Based on the above preliminary screening results, select I. 8cFurther determination of IC50 in MDA-MB-231, MCF-7, A549 / DDP, MCF-7 / DDP, and MCF-10A cells. 50 The values ​​were determined, and cisplatin (DDP) and 11 were selected as controls.

[0149] The test results are shown in Table 1. In the tested cells, compound I... 8c The activity of I was significantly better than that of cisplatin and 11, indicating that I 8c It has superior anti-tumor activity.

[0150] 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

[0151]

[0152] a ·Cells ·were ·treated ·with ·the ·indicated ·compounds ·tor ·72 ·h, ·and ·the ·cell ·viability ·and ·IC 50 ·values·were·determined·by·MTT·assay.·Data·were·expressed·as·the·mean·±·SD·from·three·individual·experiments.

[0153] b ·FI·values, ·fold·increase, ·calculated·as·IC 50 ·(DDP)· / ·IC 50 ·(I 8c ).

[0154] c ·ND: ·not·determined.

[0155] (3) Test on the binding of the drug to human serum albumin

[0156] I measured using SPR 8c The binding constant (KD) of compound 11 to human serum albumin, and I 8c Computer-generated results of binding to human serum albumin.

[0157] Using a Biacore T200 instrument (GE Healthcare) and PBS-P running buffer (10 mM phosphate buffer containing 2.7 mM KCl, 137 mM NaCl, and 0.05% surfactant P20), the temperature was 25 °C. Human serum albumin was immobilized onto the CM5 sensor chip using a standard amine coupling program (10 mM sodium acetate (pH 5.5)). The compounds under investigation were serially diluted and then passed through the sensor chip at a flow rate of 30 μL / min for 120 seconds during the contact phase, followed by a 120-second buffer injection during the dissociation phase. KD values ​​were calculated using Biacore T200 evaluation software version 1.0 (GE Healthcare).

[0158] Test results are as follows Figure 3 As shown in A, Ⅰ 8c The KD value of human serum albumin is 15.35 μM. In stark contrast, compound 11 ( Figure 3 B) is a compound analogue of patent ZL202011077271.9, which does not contain the alkyl long chain moiety and whose KD value exceeds the detection limit and cannot be determined. This highlights the key role of the long alkyl chain in the binding affinity of human serum albumin.

[0159] (4) 8c Pt uptake by DDP in MCF-7 cells and MDA-MB-231 cells.

[0160] Using the same concentration (1 μmol / L) of I 8c Incubate cisplatin with MCF-7 cells and MDA-MB-231 cells for 24 hours (e.g.) Figure 4 As shown in A), 48h (as shown in A) Figure 4 (As shown in B), and then the Pt uptake of different cells was observed by ICP-MS. The test results are as follows. Figure 4 As shown.

[0161] Depend on Figure 4 It can be seen that compound I is present in all cells. 8c All of these are much higher than the uptake of cisplatin (Pt). Part of the reason is compound I. 8c Its lipid solubility and stability are much higher than cisplatin. On the other hand, this may be due to I... 8c The released NO caused nitrosation of metal transporters Atox1 and ATP7a and reduced their activity, resulting in a decrease in the efflux of platinum by metal transporters in tumor cells.

[0162] (5) Compound I 8c The intracellular NO release status.

[0163] Detection of compound I using DAF-FM DA fluorescent probe 8c The results of flow cytometry analysis of NO release in different cells are as follows: Figure 5 As shown.

[0164] Depend on Figure 5 We can obtain, Ⅰ 8c NO release in tumor cells MDA-MB-231 and MCF-7 / DDP was greater than that in normal breast epithelial cells MCF-10A, indicating that compound I... 8c It selectively degrades and catalyzes the release of NO in tumor cells, and exhibits good biocompatibility with normal cells.

[0165] (6) Compound I 8c Effects on the migration ability of MDA-MB-231 cells

[0166] Next, we will study compound I. 8c To investigate the effect of the compound's antiproliferative activity on the migration ability of MDA-MB-231 cells, and to prevent this from significantly impacting migration ability, the IC50 of the compound on tumor cells was first calculated, referring to the aforementioned method. 50 The method for calculating the IC50 value (MTT method) first tests and calculates the IC50 of the compound. 10 The value is 84.477 nM.

[0167] Secondly, the transwell migration experiment demonstrated the efficacy of compound I. 8c It can significantly inhibit the migration ability of MDA-MB-231 cells. The transwell migration assay measures cell migration ability by counting the number of migrating cells; a higher cell count indicates stronger migration ability. Figure 6 As shown in Figure A, the experimental results indicate that compound I 8c It can significantly inhibit the migration ability of MDA-MB-231 cells.

[0168] Then we continued using 85nM I 8c The scratch test was conducted, and the results were as follows: Figure 6 As shown in B, compound I 8c The scratch closure degree of I was lower than that of the control group, indicating that 8c It significantly inhibited the migration ability of MDA-MB-231 cells.

[0169] (7) Compound I 8c Metabolic properties in the body

[0170] To further evaluate compound I 8c The metabolic properties of compound I in vivo were determined. 8cThe pharmacokinetic (PK) properties of the active ingredient and total platinum in rats were investigated. Three male SD rats were selected and administered I via intravenous injection. 8c (5 mg / kg) Blood samples were collected from the fundus venous plexus before administration and at 5 min, 15 min, 30 min, 60 min, 2 h, 4 h, 6 h, 8 h, and 24 h after administration. The supernatant plasma was collected by centrifugation and stored at -20℃ for later use. The test results are shown in Table 2.

[0171] Tabie·2.·PK·Parameters·of·I 8c (iv·5·mg / kg). a

[0172]

[0173] a ·Values·are·the·average·of·three·determinations.

[0174] From Table 2, we can see that I 8c The half-life in vivo is 0.29 h, T max Its AUC is 0.08h. (0-t) The concentration was 2210.10 ug / L*h. The total platinum half-life in rats was 23.58 h, and T... max Its AUC is 0.08h. (0-t) It is 14720.28ug / L*h.

[0175] (8) Compound I 8c Acute toxicity

[0176] Compound I was performed 8c Acute toxicity test, observe compound I 8c Acute toxicity was assessed to determine a safe dosage range. Preliminary experiments were conducted, showing that the test drug had some toxicity. Intravenous injection of 70 mg / kg caused death in 4 / 4 mice, while intravenous injection of 30 mg / kg caused death in 0 / 4 mice. Based on the results of the preliminary experiments, we selected the following doses (63 mg / ml, 56.7 mg / ml, 51.03 mg / ml, 45.93 mg / ml, and 41.34 mg / ml) for LD50 testing. 50 Experiment. The drug was administered intravenously once at the above dosage. The symptoms of poisoning and mortality of mice in each group were recorded, and autopsies were performed on the dead animals.

[0177] The experimental results are as follows:

[0178] Abnormal reactions: After intravenous injection of the drug into mice, animals in all dosage groups exhibited lethargy, paralysis, and death. The higher the dosage, the more pronounced the symptoms and the shorter the time to death. These experimental results indicate that compound I... 8c It is somewhat toxic to mice; administration of higher doses can cause death in mice. 8c LD50 after intravenous administration 50 The value was 53.2507 (48.3854~58.6053) mg / kg.

[0179] (9) Compound I 8c Efficacy evaluation of MDA-MB-23l in vivo xenograft tumors in mice

[0180] The efficacy of MDA-MB-231 cells in in vivo xenograft tumors in mice was tested. MDA-MB-231 cells were inoculated into the second pair of mammary pads on the right side of mice, and tumors were established and grown to 100 mm. 3 Mice were randomly divided into three groups (left and right). 8c The treatment group, a negative control group receiving only solvent, and a DDP group were included. The experimental groups received different doses of I... 8c (5, 2.5 and 1.25 mg / kg, intravenously every three days), DDP group: 5 mg / kg, intravenously every three days.

[0181] The results are as follows Figure 7 As shown. Figure 7 A represents the results of tumor volume measurement and calculation every other day; Figure 7 B represents the mouse's weight measured every other day; Figure 7 C represents the tumor weight measurement result in mice; Figure 7 D is an image of a mouse tumor. From the image, we can see that compound I... 8c Dose-dependent inhibition of tumor cell growth, of which I 8c The tumor inhibition rate of the cisplatin group (5 mg / kg) was 71.08%, while the tumor inhibition rate of the cisplatin group (5 mg / kg) was only 58.51%. 8c Its in vivo pharmacodynamic activity was significantly superior to cisplatin. Furthermore, I 8c The high, medium, and low dose groups all had a lower effect on mouse body weight than the cisplatin group, indicating that compound I... 8c It has better safety compared to cisplatin.

[0182] (10) Compound I 8c In vivo anti-MDA-MB-231-luc lung metastasis

[0183] For I 8cThe lung metastasis activity of anti-MDA-MB-231 cells in mice was tested in vivo. Six-week-old female BALB / c mice were administered MDA-MB-231 / luc cells (5 × 10⁻⁶) via tail vein. 7 (cells). One week later, the animals were randomly divided into three groups of six mice each. Compound I 8c The treatment group received a tail vein injection of 2.5 mg / kg every three days. The DDP treatment group also received a tail vein injection of 2.5 mg / kg every three days. The negative control group received the solvent at the same frequency. Mice were anesthetized with 2% inhaled isoflurane and intraperitoneally injected with D-fluorescein for imaging. Signals were acquired using the IVIS Lumina imaging system 10 minutes later. Test results are as follows: Figure 8 As shown; where Figure 8 A consists of three representative fluorescence imaging images of small animals; Figure 8 B is a statistical graph showing the quantitative calculation of fluorescence intensity in the lungs of mice in each group.

[0184] Depend on Figure 8 It can be seen that, compared with the control group and the DDP group, using I 8c The treated mice showed significantly lower fluorescence intensity on day 12, indicating that I 8c The treatment significantly inhibited lung metastases of MDA-MB-231 in vivo.

[0185] (11)Ⅰ 8c Tests to detect changes in intracellular Atox1 and ATP7a nitrosation levels in tumor cells

[0186] Compound I 8c The cells were incubated with MDA-MB-231 cells, and the nitrite levels of Atox1 and ATP7a were then detected using the Biotin-Switch method.

[0187] Test results are as follows Figure 9 As shown, where, Figure 9 A is to verify I using the Biotin-Switch method. 8c The experimental results showing that the released NO caused an increase in Atox1 and ATP7a nitrosation in MDA-MB-231 cells; Figure 9 B is its strip grayscale statistical graph. From the graph, it can be seen that compared to DDP, I... 8c Released NO significantly increased S-nitrosolysis by Atox1 and ATP7a. Increased nitrosolysis by these two transporters led to decreased metal-binding activity and increased platinum retention in cancer cells, thereby enhancing the integration of prodrug I. 8c It has a more effective anti-tumor effect.

Claims

1. A nitric oxide donor-type tetravalent platinum prodrug, characterized in that, Its structural formula is: or ; where R2 is selected from C4~C 18 alkyl.

2. The nitric oxide donor-type tetravalent platinum prodrug according to claim 1, characterized in that, R2 is selected from C6~C 12 alkyl.

3. The nitric oxide donor-type tetravalent platinum prodrug according to claim 1, characterized in that, R2 is a dodecyl group.

4. The nitric oxide donor-type tetravalent platinum prodrug according to claim 1, characterized in that, -(H3N)2Cl2 on Pt is cis.

5. A method for preparing a nitric oxide donor-type tetravalent platinum prodrug according to claim 1, characterized in that, The structural formula of the nitric oxide donor-type tetravalent platinum prodrug is: The steps include: (1) Compound I4 undergoes an amide condensation reaction with succinic anhydride to give compound I5; (2) Compound I5 undergoes an esterification reaction with N-hydroxysuccinimide to give compound I6; (3) Compound I6 undergoes an ester exchange reaction with cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum to give compound I7; (4) Compound I7 undergoes an amination reaction with R2-N=C=O to give compound I8; The synthesis route is as follows: ; where R2 is selected from C4~C 18 alkyl.

6. A method for preparing a nitric oxide donor-type tetravalent platinum prodrug according to claim 1, characterized in that, The structural formula of the nitric oxide donor-type tetravalent platinum prodrug is: The steps include: (1) Compound II3 undergoes an amide condensation reaction with succinic anhydride to give compound II4; (2) Compound II4 undergoes esterification with N-hydroxysuccinimide to give compound II5; (3) Compound II5 undergoes an ester exchange reaction with cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum to give compound II6; (4) Compound II6 undergoes an amination reaction with R2-N=C=O to give compound II7; The synthesis route is as follows: ; where R2 is selected from C4~C 18 alkyl.

7. The method for preparing a nitric oxide donor-type tetravalent platinum prodrug according to claim 5 or 6, characterized in that, In the transesterification reaction of compound I6 or II5 with cis-diamine dichlorodihydroxyplatinum or trans-diamine dichlorodihydroxyplatinum, the solvent used is anhydrous DMSO, the reaction conditions are to avoid light, and the reaction temperature is 75~80℃.

8. A pharmaceutical composition, characterized in that, The product comprises a tetravalent platinum prodrug containing a nitric oxide donor as described in any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

9. The use of the nitric oxide donor-type tetravalent platinum prodrug according to any one of claims 1 to 4 in the preparation of antitumor drugs for prevention or treatment.

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