Febuxostat tetravalent platinum prodrug, preparation method and preparation

Abstract

The present invention belongs to the field of pharmaceutical technology, and discloses a febuxostat tetravalent platinum prodrug, a preparation method and a preparation. The present invention synthesizes two small molecule tetravalent platinum prodrugs of tetravalent platinum, febuxostat cisplatin and febuxostat oxaliplatin, for the first time, and prepares a febuxostat cisplatin nanoformulation. The nanoformulation solves the shortcomings of cisplatin such as poor water solubility, congenital or acquired drug resistance, and obvious toxic and side effects, and has many advantages such as passive targeting of tumor tissue. It is not only economical and practical, but also provides possibilities for industrial production. By using the idea of ​​"drug repositioning", the anti-tumor effect of febuxostat is further studied, and a combined anti-tumor effect with platinum drugs is achieved. At the same time, a carrier-free preparation method is adopted to avoid the unknown toxicity of the carrier, the preparation has good safety, and the tumor targeting of platinum drugs is achieved by nanoformulation, while enhancing the accumulation of tumor sites and reducing systemic toxic and side effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and in particular relates to a febuxostat tetravalent platinum prodrug, a preparation method and a preparation thereof. Background Art

[0002] Chemotherapy is a common treatment for cancer. In clinical practice, platinum-based drugs are among the most widely used broad-spectrum anticancer agents, including the first-generation platinum compound cisplatin (CDDP), the second-generation platinum compound carboplatin (CBP), and the third-generation platinum compound oxaliplatin (L-OHP). Cisplatin, the first approved platinum drug, has expanded its application from testicular cancer to various solid tumors, including bladder cancer, ovarian cancer, lung cancer, and head and neck cancer. Platinum-based drugs target DNA, affecting its normal replication and transcription, leading to cell apoptosis or necrosis, achieving antitumor effects. While platinum-based drugs offer advantages such as simple structure and robust efficacy, they also suffer from poor water solubility, congenital or acquired drug resistance, and significant toxic side effects, often failing to induce effective apoptosis. Therefore, enhancing the antitumor activity and mitigating their toxic side effects remain hot topics in research.

[0003] Studies have shown that the conversion of divalent platinum (Pt(II)) to tetravalent platinum (Pt(IV)) prodrugs can effectively improve the therapeutic efficacy of platinum-based anticancer agents. After entering tumor cells, the Pt(IV) prodrug decomposes into Pt(II) and an axially active ligand under the action of intracellular reducing substances, and the two synergistically exert antitumor activity. Compared with Pt(II), the Pt(IV) prodrug has a regular octahedral configuration that is more kinetically inert and can reduce toxic side effects. In addition, the axial position of the Pt(IV) prodrug is more susceptible to chemical modification, so it can be given the desired pharmacological effect by introducing different active groups at the axial position.

[0004] The Chinese patent for the prior art: CN112961188B, publication date: June 15, 2021, discloses a tetravalent platinum prodrug benzyldalate platinum, its preparation, preparation method and application. The Chinese patent for the prior art: CN112961190A, publication date: June 15, 2021, discloses a tetravalent platinum veratrate and its preparation method and application. In the above-mentioned prior art, only one preparation method is proposed. Febuxostat (FBX) is a uric acid-lowering drug, and the present invention can selectively inhibit xanthine oxidase. This type of inhibitor is an axial group of tetravalent platinum, which is not recorded in the above-mentioned technical solution.

[0005] The effectiveness of FBX in preventing cell lysis syndrome in patients with malignant tumors has also been recognized. In recent years, the application of FBX in anti-tumor treatment has also been widely studied. Based on the above research, it is planned to "drug repositioning" FBX, that is, to explore new uses for molecules with known pharmacological effects. The carboxyl functional group contained in the chemical structure of FBX is conducive to its further chemical modification and can be used as a modified ligand for Pt(IV) prodrugs. The structural formula of Febuxostat is:

[0006]

[0007] Although Pt(IV) prodrugs can reduce the toxicity of divalent platinum to a certain extent, they still have the disadvantages of small molecule drugs, such as short blood circulation time, poor accumulation ability in tumor sites, and lack of tumor targeting. More and more studies have shown that nanodelivery systems can effectively improve the therapeutic effect of platinum drugs. Due to the complexity of tumors, the gaps between blood vessels in tumor tissues are wide and the structural integrity is poor, which makes nanoparticles of appropriate size (20-200nm) have selective high permeability and retention, namely the EPR effect. Therefore, nanodelivery systems have the following advantages: (1) achieving the delivery of poorly soluble drugs and improving their bioavailability; (2) passively targeting to tumor sites through the EPR effect, reducing systemic toxic side effects; (3) preventing rapid drug degradation and improving stability; (4) promoting drug endocytosis and improving intracellular penetration.

[0008] Through the above analysis, the problems and defects of the existing technology are as follows:

[0009] (1) The tetravalent platinum prodrugs provided in the prior art have poor passive targeting effects on tumor tissues.

[0010] (2) The tetravalent platinum prodrugs and platinum drugs provided in the prior art have poor safety in the process of anti-tumor use. Summary of the Invention

[0011] To overcome the problems existing in the related art, the embodiments disclosed in the present invention provide a febuxostat tetravalent platinum prodrug, a preparation method and a preparation.

[0012] The technical solution is as follows: a febuxostat tetravalent platinum prodrug, wherein the febuxostat tetravalent platinum prodrug includes febuxostat-cisplatin;

[0013] The structural formula of Febuxostat-Cisplatin is:

[0014]

[0015] Another object of the present invention is to provide a method for preparing a febuxostat tetravalent platinum prodrug, comprising the following steps:

[0016] S1. Synthesis of hydroxyplatin: Weigh cisplatin, slowly add hydrogen peroxide, protect from light, and heat under reflux at 40-100°C for 1-24 hours. After the reaction is completed, stand at 0-10°C for crystallization, centrifuge, and wash the solid after centrifugation twice with distilled water, ethanol, and ether, respectively, and dry to obtain a light yellow solid hydroxyplatin;

[0017] S2, Synthesis of Febuxostat tetravalent platinum prodrug: Weigh the hydroxyplatinum, add DMF solvent, then add Febuxostat, triethylamine and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) in sequence, protect from light, protect under nitrogen, stir at 50°C until the reaction solution is clear, add ice water to the reaction solution to precipitate;

[0018] S3, purification of febuxostat platinum-IV using two purification methods;

[0019] Method 1: The precipitate is dissolved in methanol, purified by silica gel column, and gradient eluted with a mixture of dichloromethane and methanol to obtain solid febuxostat tetravalent platinum;

[0020] Method 2: The precipitate is washed with saturated potassium carbonate, distilled water, and methanol respectively to obtain solid febuxostat tetravalent platinum.

[0021] Furthermore, in step S1, the molar ratio of the cisplatin to the hydrogen peroxide is 1:(5-200), and the mass concentration of the hydrogen peroxide is 30%.

[0022] Furthermore, the synthetic chemical reaction formula of hydroxyplatinum in step S1 is:

[0023]

[0024] Furthermore, in step S2, the synthetic chemical reaction formula of febuxostat tetravalent platinum prodrug is:

[0025]

[0026] Furthermore, in step S2, the molar ratio of hydroxyplatinum: febuxostat: triethylamine: TBTU is (1-3):(3-9):(3-9):(3-9); the stirring time at 50° C. is 12-96 hours;

[0027] In step S3, the volume ratio of dichloromethane to methanol in method 1 is 20:1; the saturated potassium carbonate and distilled water in method 2 are each washed twice, and the methanol is washed once.

[0028] Another object of the present invention is to provide a method for preparing a tetravalent platinum prodrug molecule febuxostatin preparation, comprising the following steps:

[0029] (1) dissolving febuxostat-cisplatin in a febuxostat tetravalent platinum prodrug in DMSO to form a febuxostat-cisplatin solution;

[0030] (2) Deionized water was added to a vial equipped with a magnetic rod, and the above-mentioned Febuxostat-cisplatin DMSO solution was slowly added dropwise during stirring. Stirring was continued for 20 minutes. After stirring, the solution was centrifuged and the supernatant was discarded. Water was added to suspend the solution, and the supernatant was discarded repeatedly. The precipitate was completely dispersed by ultrasonication with 1 ml of deionized water to obtain the Febuxostat-cisplatin nanoformulation.

[0031] Furthermore, the concentration of febuxostat-cisplatin is 10 mg / ml.

[0032] Another object of the present invention is to provide a platinum prodrug molecule febuxostatin preparation, which is prepared according to the preparation method of the tetravalent platinum prodrug molecule febuxostatin preparation.

[0033] Another object of the present invention is to provide a febuxostat tetravalent platinum prodrug, wherein the febuxostat tetravalent platinum prodrug further comprises febuxostat-oxaliplatin;

[0034] The structural formula of Febuxostat-Oxaliplatin is:

[0035]

[0036] Combining all the above technical solutions, the advantages and positive effects of the present invention are as follows: the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation provided by the present invention, this small molecule solves the shortcomings of cisplatin such as poor water solubility, congenital or acquired drug resistance, and obvious toxic and side effects.

[0037] The present invention can synthesize the tetravalent platinum prodrug febuxostat-cisplatin through a simple and rapid method, and provides a simple, feasible, environmentally friendly, and relatively high-yield purification method. At the same time, a structurally uniform nanoformulation is formed under the action of self-assembly, which has many advantages such as passive targeting of tumor tissues. It is not only economical and practical, but also provides the possibility of industrial production.

[0038] After intravenous injection, the tetravalent platinum prodrug febuxostat-cisplatin nanoformulation can passively target the tumor site and increase the tumor cells' ability to uptake the drug. The results of cell experiments show that the anti-tumor effect is far superior to cisplatin and its combination drug.

[0039] The quadrivalent platinum prodrug febuxostat-cisplatin and its nanoformulation use the concept of "drug repositioning" to further study the anti-tumor effect of febuxostat and achieve combined anti-tumor effects with platinum drugs. At the same time, the use of carrier-free formulations avoids the unknown toxicity of the carrier, and the preparation has good safety. The tumor targeting of platinum drugs is achieved through nanoformulations, enhancing accumulation in tumor sites while reducing systemic toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present disclosure and, together with the description, serve to explain the principles of the present disclosure;

[0041] Figure 1 1 is a flow chart of a method for preparing a febuxostat tetravalent platinum prodrug provided in an embodiment of the present invention;

[0042] Figure 2 This is the hydrogen nuclear magnetic resonance spectrum of the tetravalent platinum prodrug febuxostat-cisplatin provided in an embodiment of the present invention;

[0043] Figure 3 The hydrogen nuclear magnetic resonance spectrum of the tetravalent platinum prodrug febuxostat-oxaliplatin provided in an embodiment of the present invention;

[0044] Figure 4 is the mass spectrum of the tetravalent platinum prodrug febuxostat-cisplatin provided in an embodiment of the present invention;

[0045] Figure 5 This is the mass spectrum of the tetravalent platinum prodrug febuxostat-oxaliplatin provided by an embodiment of the present invention.

[0046] Figure 6 This is a TEM image of the particle morphology of the tetravalent platinum prodrug febuxostat-cisplatin nanoformulation provided in an embodiment of the present invention;

[0047] Figure 7 This is a particle size distribution diagram of the tetravalent platinum prodrug febuxostat-cisplatin nanoparticle preparation provided by an embodiment of the present invention;

[0048] Figure 8 1 is a zeta potential diagram of a tetravalent platinum prodrug febuxostat-cisplatin nanoformulation provided in an embodiment of the present invention;

[0049] Figure 9 Figure 3: Antitumor effect of the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation on A549 cells provided by the embodiments of the present invention;

[0050] Figure 10 Figure 3: Antitumor effect of the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation on FaDu cells provided by the embodiments of the present invention;

[0051] Figure 11 Figure 3: Anti-tumor effect of the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation on HeLa cells provided by the embodiments of the present invention;

[0052] Figure 12 Figure 3: Antitumor effect of the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation on MDA-MB-231 cells provided by the embodiments of the present invention;

[0053] Figure 13 Figure 3: Antitumor effect of the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation on HCT116 cells provided by the embodiments of the present invention;

[0054] Figure 14 Figure 3 shows the anti-tumor effect of the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation on PLC / PRF / 5 cells provided by an embodiment of the present invention. DETAILED DESCRIPTION

[0055] To make the above-mentioned objects, features, and advantages of the present invention more readily apparent, specific embodiments of the present invention are described in detail below with reference to the accompanying drawings. The following description sets forth numerous specific details to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways than those described herein, and those skilled in the art may make similar modifications without departing from the scope of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0056] Example 1: This embodiment of the present invention provides a febuxostat tetravalent platinum prodrug, the structural formula of which is shown below:

[0057] Febuxostat-cisplatin:

[0058]

[0059] or febuxostat-oxaliplatin:

[0060]

[0061] Example 2, taking cisplatin as an example, describes the preparation method of febuxostat tetravalent platinum prodrug. Figure 1 As shown, the embodiment of the present invention provides a method for preparing a febuxostat tetravalent platinum prodrug, comprising the following steps:

[0062] S1. Synthesis of hydroxyplatin: Weigh cisplatin, slowly add hydrogen peroxide, protect from light, and heat under reflux at 40-100°C for 1-24 hours. After the reaction is completed, stand at 0-10°C for crystallization, centrifuge, and wash the solid after centrifugation twice with distilled water, ethanol, and ether, respectively, and dry to obtain a light yellow solid hydroxyplatin;

[0063] The chemical reaction formula is:

[0064]

[0065] S2, Synthesis of Febuxostat tetravalent platinum prodrug: Weigh the hydroxyplatinum, add DMF solvent, then add Febuxostat, triethylamine and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) in sequence, protect from light, protect under nitrogen, stir at 50°C until the reaction solution is clear, add ice water to the reaction solution to precipitate;

[0066] The chemical reaction formula is:

[0067]

[0068] S3, purification of febuxostat platinum-IV using two purification methods;

[0069] Method 1: The precipitate is dissolved in methanol, purified by silica gel column, and gradient eluted with a mixture of dichloromethane and methanol to obtain solid febuxostat tetravalent platinum;

[0070] Method 2: The precipitate is washed with saturated potassium carbonate, distilled water, and methanol respectively to obtain solid febuxostat tetravalent platinum.

[0071] In an embodiment of the present invention, in step S1, the molar ratio of cisplatin to hydrogen peroxide is 1:(5-200), and the mass concentration of hydrogen peroxide is 30%.

[0072] The temperature of the heating reflux is preferably 70°C, the time is preferably 5 hours, and the temperature of the crystallization is preferably 4°C.

[0073] In an embodiment of the present invention, in step S2, the molar ratio of hydroxyplatinum: febuxostat: triethylamine: TBTU is (1-3): (3-9): (3-9): (3-9); preferably, the molar ratio of hydroxyplatinum: febuxostat: triethylamine: TBTU is 1:3:3:3; and the stirring time at 50° C. is 12-96 hours, preferably 72 hours.

[0074] In an embodiment of the present invention, in step S3, the volume ratio of dichloromethane to methanol in method 1 is 20:1; the saturated potassium carbonate and distilled water in method 2 are each washed twice, and the methanol is washed once.

[0075] Example 3, the present invention provides a method for preparing a tetravalent platinum prodrug molecule febuxostatin preparation, comprising the following steps:

[0076] (1) dissolving febuxostat-cisplatin in DMSO to form a febuxostat-cisplatin solution;

[0077] (2) Add deionized water to a vial equipped with a magnetic rod. Slowly add the above-mentioned febuxostat-cisplatin DMSO solution dropwise during stirring. Continue stirring for 20 minutes until an opalescent effect is observed. After stirring, centrifuge and gently discard the supernatant. Add water to resuspend, repeat centrifugation, and discard the supernatant. Ultrasonicate the precipitate with 1 ml of deionized water to completely disperse it, thereby obtaining the febuxostat-cisplatin nanoformulation.

[0078] As an improvement, the concentration of febuxostat-cisplatin is 10 mg / ml.

[0079] As an improvement, the febuxostat-cisplatin preparation is a nano preparation.

[0080] The following embodiments are provided below taking cisplatin as an example.

[0081] Example 4: 200 mg (0.67 mmol) of cisplatin was accurately weighed into a round-bottom flask using an analytical balance. 13 mL of 30 wt% hydrogen peroxide was slowly added. The round-bottom flask was protected from light with tin foil and refluxed at 40°C for 10 hours. After the reaction was complete, the round-bottom flask was placed in a 0°C refrigerator for crystallization. The mixture was filtered using a Buchner funnel to obtain a solid. The solid was washed twice with distilled water, ethanol, and ether. The solid was dried in a vacuum drying oven overnight to obtain a light yellow solid hydroxyplatinum.

[0082] 100 mg (0.3 mmol) of hydroxyplatinum was accurately weighed into a round-bottom flask using an analytical balance. 2 mL of anhydrous DMF was added. 284.74 mg (0.9 mmol) of febuxostat, 130 μL (0.9 mmol) of triethylamine, and 288 mg (0.9 mmol) of TBTU were accurately weighed and added to the round-bottom flask. The round-bottom flask was protected from light with tin foil, evacuated, and replaced with a nitrogen balloon three times. Stir at 50°C for 24 hours until the reaction solution became clear. Ice water was added to the reaction solution to precipitate the precipitate, which was re-dissolved in methanol and mixed with silica gel. The evaporated solid was purified by silica gel column chromatography using a mixture of dichloromethane and methanol as the eluent (a volume ratio of dichloromethane to methanol of 20:1). The product was collected and dried to obtain febuxostatin as a yellow solid with a yield of 38.1%.

[0083] Example 5. 200 mg (0.67 mmol) of cisplatin was accurately weighed into a round-bottom flask using an analytical balance. 0.35 mL of 30 wt% hydrogen peroxide was slowly added. The round-bottom flask was protected from light with tin foil and refluxed at 100° C. for 1 hour. After the reaction was complete, the round-bottom flask was placed in a 10° C. refrigerator for crystallization. The mixture was filtered using a Buchner funnel to obtain a solid. The solid was washed twice with distilled water, ethanol, and ether. The solid was dried in a vacuum drying oven overnight to obtain a light yellow solid hydroxyplatinum.

[0084] 100 mg (0.3 mmol) of hydroxyplatinum was accurately weighed into a round-bottom flask using an analytical balance. 2 mL of anhydrous DMF was added. 854.23 mg (2.7 mmol) of febuxostat, 390 μL (2.7 mmol) of triethylamine, and 864 mg (2.7 mmol) of TBTU were accurately weighed and added to the round-bottom flask. The round-bottom flask was protected from light with tin foil, evacuated, and replaced with a nitrogen balloon three times. Stir at 50°C for 48 hours until the reaction solution became clear. Ice water was added to the reaction solution to precipitate the precipitate, which was re-dissolved in methanol and mixed with silica gel. The evaporated solid was purified by silica gel column chromatography using a mixture of dichloromethane and methanol as the eluent (a volume ratio of dichloromethane to methanol of 20:1). The product was collected and dried to obtain febuxostatin as a yellow solid with a yield of 35.2%.

[0085] Example 6: 200 mg (0.67 mmol) of cisplatin was accurately weighed into a round-bottom flask using an analytical balance. 3 mL of 30 wt% hydrogen peroxide was slowly added. The round-bottom flask was protected from light with tin foil and refluxed at 70°C for 5 hours. After the reaction was complete, the round-bottom flask was placed in a 4°C refrigerator for crystallization. The mixture was filtered using a Buchner funnel to obtain a solid. The solid was washed twice with distilled water, ethanol, and ether. The solid was dried in a vacuum drying oven overnight to obtain a light yellow solid hydroxyplatinum.

[0086] 100 mg (0.3 mmol) of hydroxyplatinum was accurately weighed on an analytical balance in a round-bottom flask, 2 mL of anhydrous DMF solvent was added, and 284.74 mg (0.9 mmol) of febuxostat, 130 μL (0.9 mmol) of triethylamine, and 288 mg (0.9 mmol) of TBTU were accurately weighed and added to the round-bottom flask. The round-bottom flask was protected from light with tin foil, evacuated, replaced with a nitrogen balloon three times and protected, and stirred at 50 ° C for 72 hours until the reaction solution became clear. Ice water was added to the reaction solution to precipitate, and the precipitate was washed twice with saturated potassium carbonate, distilled water, and methanol, and then dried in vacuo to obtain yellow solid febuxostatin with a yield of 66.2%.

[0087] Example 7, Preparation of tetravalent platinum prodrug febuxostat-cisplatin nanoformulation:

[0088] Dissolve 4 mg of febuxostat-cisplatin in 400 μl of DMSO to form a febuxostat-cisplatin solution. Add 4 ml of deionized water to a vial equipped with a magnetic stirrer and slowly add the febuxostat-cisplatin solution from step 1 dropwise. Continue stirring for 20 minutes. After stirring, centrifuge and discard the supernatant. Add another 4 ml of deionized water, remix, centrifuge, and discard the supernatant. Add 1 ml of ultrapure water and mix thoroughly to obtain the febuxostat-cisplatin nanoformulation.

[0089] The present invention analyzes the tetravalent platinum prodrug Febuxostat-cisplatin nanoformulation: Take a drop of the tetravalent platinum prodrug Febuxostat-cisplatin nanoformulation prepared in Example 7 and drop it on the carbon film copper mesh. The excess liquid is absorbed by filter paper and the aggregate morphology of Febuxostat-cisplatin is observed under a transmission electron microscope after drying. Figure 6 As shown, the results showed that febuxostat-cisplatin could form nanoparticles of uniform size and good dispersion.

[0090] The particle size distribution analysis of the tetravalent platinum prodrug febuxostat-cisplatin nanoparticle preparation in the present embodiment: The tetravalent platinum prodrug febuxostat-cisplatin nanoparticle preparation prepared in Example 7 was taken and the particle size and Zeta potential of the nanoparticle preparation solution were measured using a Nano-ZS90 particle size potential meter at room temperature. The results are shown in FIG. Figure 7 As shown in Figure 2, the average particle size of Febuxostat-Cisplatin nanoparticles is about 142.5nm, with uniform morphology. After intravenous injection, they can be passively targeted to tumor tissues through the EPR effect. Figure 8 The value shown is -35.5mV, which indicates good stability.

[0091] In the embodiment of the present invention, the nuclear magnetic resonance spectroscopy ( 1 H-NMR) to identify the chemical structure of Febuxostat-Cisplatin. Figure 2 shown.

[0092] About 5 mg of febuxostatin prepared in Example 4 was weighed, dissolved in deuterated dimethyl sulfoxide (DMSO-d6) and placed in a nuclear magnetic resonance tube. Its nuclear magnetic resonance 1H spectrum was measured using a 400 MHz nuclear magnetic resonance 1H spectrum with tetramethylsilane as the internal standard, and the chemical shift value (ppm) of the compound was recorded. The results are shown in FIG. Figure 2 As shown, Figure 2 The NMR results can confirm that each peak in the newly synthesized purified molecule can be assigned. 1 The H-NMR spectrum can confirm the successful synthesis of febuxostat-cisplatin.

[0093] In the embodiment of the present invention, the nuclear magnetic resonance carbon hydrogen spectrum ( 1 H-NMR) to identify the chemical structure of Febuxostat-Oxaliplatin. Figure 3 shown.

[0094] About 5 mg of febuxostatin prepared in Example 4 was weighed, dissolved in deuterated dimethyl sulfoxide (DMSO-d6) and placed in a nuclear magnetic resonance tube. Its nuclear magnetic resonance 1H spectrum was measured using a 400 MHz nuclear magnetic resonance 1H spectrum with tetramethylsilane as the internal standard, and the chemical shift value (ppm) of the compound was recorded. The results are shown in FIG. Figure 3 As shown, Figure 3 The NMR results can confirm that each peak in the newly synthesized purified molecule can be assigned. 1The H-NMR spectrum can confirm the successful synthesis of febuxostat-oxaliplatin.

[0095] In the embodiment of the present invention, the chemical structure of febuxostat-cisplatin was identified by mass spectrometry (MS), such as Figure 4 shown.

[0096] Weigh about 1 mg of febuxostatin prepared in Example 4, dissolve it in methanol, and perform mass spectrometry analysis. Figure 4 As shown, Figure 4 The mass spectrometry results show that the molecular ion peak [MH] - The appearance of (939.23) proved that the synthesis of febuxostat-cisplatin was successful.

[0097] In the embodiment of the present invention, Figure 5 As shown, the chemical structure of febuxostat-oxaliplatin was identified by mass spectrometry (MS):

[0098] Weigh about 1 mg of febuxostatin prepared in Example 4, dissolve it in methanol, and perform mass spectrometry analysis. Figure 5 As shown, Figure 5 The mass spectrometry results show that the molecular ion peak [MH] - The emergence of (1025.86.23) proved the successful synthesis of febuxostat-oxaliplatin.

[0099] In the examples of the present invention, the in vitro anti-tumor effect of febuxostatin was analyzed:

[0100] The CCK-8 method was used to perform the assay. Six tumor cell lines were selected: A549 (human non-small cell lung cancer cells), HeLa (human cervical cancer cells), FaDu (human pharyngeal squamous cells), MDA-MB-231 (human breast cancer cells), HCT116 (human colon cancer cells), and PLC / PRF / 5 (human liver cancer cells). Cells in the logarithmic growth phase were collected and the cell suspension concentration was adjusted to 5×10 3 / well were inoculated in a 96-well plate and placed in a 37°C, 5% CO2 incubator for 24 hours until completely adhered. 0-100 μM drugs were prepared using the gradient dilution method, and different concentrations of drugs were added in sequence, with 3 replicates set for each concentration. After incubation in a CO2 incubator for 48 hours, 10% CCK-8 solution was added, protected from light, and incubated for another 1 hour. The absorbance of each well was detected at a wavelength of 450 nm using a microplate reader. All values ​​were expressed as mean ± standard deviation, cell inhibition rate (%) = (ODcontrol-ODtreated) / (ODcontrol-ODblank) × 100%, and statistical analysis was performed using SPSS16.0 and GraphPadPrism5 software to fit the cell growth inhibition curve and calculate the IC 50 The result is as follows Figures 9-14As shown, the results showed that the tetravalent platinum prodrug febuxostat-cisplatin and its nanoformulation showed good inhibitory effects on various tumor cells, among which the nanoformulation had the best effect on MDA-MB-231, and its anti-tumor effect was nearly 7 times higher than that of cisplatin.

[0101] This invention synthesizes for the first time two small-molecule tetravalent platinum drugs, febuxostat-cisplatin and febuxostat-oxaliplatin, as tetravalent platinum prodrugs. It also produces a febuxostat-cisplatin nanoformulation. This nanoformulation overcomes the shortcomings of cisplatin, such as poor water solubility, congenital or acquired drug resistance, and significant toxic side effects. It offers numerous advantages, including passive tumor tissue targeting, making it economical and practical, while also facilitating industrial production. Leveraging the concept of "drug repositioning," further research has been conducted on the anti-tumor effects of febuxostat, enabling its combined anti-tumor activity with platinum drugs. Furthermore, a carrier-free formulation approach avoids the unknown toxicity of the carrier, resulting in a safer formulation. Furthermore, the nanoformulation achieves tumor targeting of the platinum drug, enhancing its accumulation at the tumor site while reducing systemic toxic side effects.

[0102] In the above embodiments, the description of each embodiment has its own focus. For parts that are not described or recorded in detail in a certain embodiment, reference can be made to the relevant description of other embodiments.

[0103] The above description is only a preferred specific implementation method of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made by any technician familiar with this technical field within the technical scope disclosed by the present invention and within the spirit and principles of the present invention should be covered by the scope of protection of the present invention.

Claims

1. A febuxostat tetravalent platinum compound, characterized in that The febuxostat tetravalent platinum compound is febuxostat-cisplatin; The structural formula of Febuxostat-Cisplatin is: 。 2. A method for preparing a febuxostat tetravalent platinum compound, for preparing the febuxostat tetravalent platinum compound according to claim 1, characterized in that: The preparation method comprises the following steps: S1. Synthesis of hydroxyplatinum: Weigh cisplatin, add hydrogen peroxide, and heat under reflux at 40-100°C for 1-24 hours in the dark. After the reaction is completed, stand at 0-10°C for crystallization, centrifuge, and wash the solid after centrifugation twice with distilled water, ethanol, and ether, respectively, and dry to obtain a light yellow solid hydroxyplatinum; S2, Synthesis of a Febuxostat tetravalent platinum compound: Weigh the hydroxyplatinum, add DMF solvent, then sequentially add Febuxostat, triethylamine, and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, protect with nitrogen under light protection, stir at 50°C until the reaction solution is clear, add ice water to the reaction solution to precipitate; S3, purification of febuxostat platinum-IV using two purification methods; Method 1: The precipitate is dissolved in methanol, purified by silica gel column, and gradient eluted with a mixture of dichloromethane and methanol to obtain solid febuxostat tetravalent platinum; Method 2: The precipitate is washed with saturated potassium carbonate, distilled water, and methanol respectively to obtain solid febuxostat tetravalent platinum.

3. The method for preparing the febuxostat tetravalent platinum compound according to claim 2, wherein: In step S1, the molar ratio of the cisplatin to the hydrogen peroxide is 1:(5-200), and the mass concentration of the hydrogen peroxide is 30%.

4. The method for preparing the febuxostat tetravalent platinum compound according to claim 2, wherein In step S1, the synthetic chemical reaction formula of hydroxyplatinum is: 。 5. The method for preparing the febuxostat tetravalent platinum compound according to claim 2, wherein: In step S2, the synthetic chemical reaction formula of the febuxostat tetravalent platinum compound is: 。 6. The method for preparing the febuxostat tetravalent platinum compound according to claim 2, wherein: In step S2, the molar ratio of hydroxyplatinum, febuxostat, triethylamine, and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid is: (1-3): (3-9): (3-9): (3-9), and the stirring time at 50° C. is 12-96 hours; In step S3, the volume ratio of the dichloromethane to the methanol in method 1 is 20:1; The saturated potassium carbonate and distilled water described in method 2 were washed twice each, and methanol was washed once.

7. A method for preparing a febuxostat tetravalent platinum compound molecule febuxostat platinum preparation, characterized in that: The preparation is carried out using the febuxostat tetravalent platinum compound according to claim 1, and the preparation method comprises the following steps: (1) Dissolving febuxostat-cisplatin in DMSO to form a febuxostat-cisplatin DMSO solution; (2) Add deionized water to a vial containing a magnetic rod, slowly add the above-mentioned Febuxostat-cisplatin DMSO solution dropwise during stirring, continue stirring for 20 minutes, centrifuge after stirring, and discard the supernatant; add water to suspend, repeat centrifugation and discard the supernatant, and use 1 ml of deionized water to ultrasonically disperse the precipitate to obtain the Febuxostat-cisplatin nanoformulation.

8. The method for preparing the febuxostat tetravalent platinum compound molecule febuxostat platinum preparation according to claim 7, characterized in that: The concentration of febuxostat-cisplatin is 10 mg / ml.

9. A febuxostat tetravalent platinum compound molecule febuxostat platinum preparation, characterized in that: The method for preparing the febuxostat tetravalent platinum compound molecule febuxostat platinum preparation according to claim 7 is used.

10. The febuxostat tetravalent platinum compound molecule febuxostat platinum preparation according to claim 9, characterized in that: The febuxostatin preparation also includes febuxostat-oxaliplatin; The structural formula of Febuxostat-Oxaliplatin is: 。

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