Resveratrol-succinyl glycan-tetravalent platinum nano-drug as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medical preparations, and particularly relates to a resveratrol-succinyl glycan-tetravalent platinum nano-drug as well as a preparation method and application of the resveratrol-succinyl glycan-tetravalent platinum nano-drug. In the resveratrol-succinyl glycan-tetravalent platinum nano-drug provided by the invention, the succinyl glycan (RIC) can exert the anti-tumor activity by improving the immune function of a body; resveratrol (RES) can be used for directly acting on related targets and the like to exert anti-tumor activity by inhibiting proliferation of tumor cells, inducing apoptosis of the tumor cells, inhibiting invasion and metastasis of the tumor cells and regulating the immune function of a body. According to the invention, the succinylglycan-tetravalent platinum conjugate is used for wrapping an anti-cancer drug micromolecule RES, so that the anti-cancer curative effect of the drug is improved, the problems of toxic and side effects of platinum drugs, drug resistance of tumor cells and the like are solved, and a new idea is provided for construction and application of platinum nano-drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of medical preparations, and specifically relates to a resveratrol@succinoglycan-quadrivalent platinum nanomedicine and a preparation method and application thereof. Background Art

[0002] Platinum drugs are a class of metal complex drugs containing heavy metal platinum. They are one of the common tumor chemotherapy drugs and are widely used in the treatment of various malignant tumors such as lung cancer, bladder cancer, ovarian cancer, cervical cancer, esophageal cancer, gastric cancer, colorectal cancer, and head and neck tumors. However, the toxic side effects of platinum drugs are a major challenge in their clinical application. These side effects include nephrotoxicity, ototoxicity, neurotoxicity, gastrointestinal toxicity, and bone marrow suppression.

[0003] Nano drug systems can selectively recognize receptors that are highly expressed in tumor cells, thereby improving efficacy and reducing toxicity. They have received attention and research from relevant researchers in cancer treatment and have become an effective way to detoxify drugs and treat tumors. Nano drug carriers can use their own functional group characteristics to load drugs and form a relatively stable drug delivery system. Therefore, in order to better exert the anti-cancer effect of platinum drugs and reduce toxic side effects, it is of great significance to study new platinum nano drugs. Summary of the invention

[0004] The purpose of the present invention is to provide a resveratrol@succinoglycan-tetravalent platinum nanomedicine and a preparation method and application thereof. The resveratrol@succinoglycan-tetravalent platinum nanomedicine provided by the present invention has high inhibition on cancer cells and low toxic and side effects.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The present invention provides a resveratrol@succinoglycan-tetravalent platinum nanomedicine, comprising resveratrol and a succinoglycan-tetravalent platinum conjugate layer encapsulating the resveratrol;

[0007] The particle size of the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 500-800 nm;

[0008] The components of the succinoglycan-tetravalent platinum conjugate layer are a conjugate of a tetravalent platinum organic dianhydride complex and succinoglycan.

[0009] Preferably, the mass percentage of resveratrol in the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 2-80%.

[0010] Preferably, the mass percentage of platinum element in the resveratrol@succinoglycan-tetravalent platinum nanomedicine is 2-20%.

[0011] Preferably, the tetravalent platinum organic dianhydride complex is a tetravalent platinum-succinic anhydride complex.

[0012] The present invention also provides a method for preparing the resveratrol@succinoglycan-quadrivalent platinum nanomedicine described in the above technical solution, comprising the following steps:

[0013] The tetravalent platinum organic dianhydride complex, succinoglycan, a coupling catalyst and an organic-water mixed solvent are mixed to carry out an esterification reaction to obtain a succinoglycan-tetravalent platinum coupling product;

[0014] Mixing the succinoglycan-quadrivalent platinum conjugate, resveratrol and an organic solvent to obtain a drug-containing organic phase;

[0015] The drug-containing organic phase and water are mixed to obtain the resveratrol@succinoglycan-tetravalent platinum nanomedicine.

[0016] Preferably, the mass ratio of the tetravalent platinum organic dianhydride complex to succinopolysaccharide is 1:1-5.

[0017] Preferably, the coupling catalyst comprises 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0018] Preferably, the temperature of the esterification reaction is 15 to 50° C., and the reaction time is 10 to 36 hours.

[0019] Preferably, the mass ratio of the succinoglycan-quadrivalent platinum conjugate to resveratrol is 10:1-8.

[0020] The present invention also provides the use of the resveratrol@succinoglycan-tetravalent platinum nanomedicine described in the above technical solution or the resveratrol@succinoglycan-tetravalent platinum nanomedicine obtained by the preparation method described in the above technical solution in the preparation of anti-tumor preparations.

[0021] The present invention provides a resveratrol@succinoglycan-quaternary platinum nanomedicine, comprising resveratrol and a succinoglycan-quaternary platinum conjugate layer encapsulating the resveratrol; the particle size of the resveratrol@succinoglycan-quaternary platinum nanomedicine is 500-800nm; the composition of the succinoglycan-quaternary platinum conjugate layer is a conjugate of a tetravalent platinum organic dianhydride complex and succinoglycan; the composition of the succinoglycan-quaternary platinum conjugate layer is a conjugate of a tetravalent platinum organic dianhydride complex and succinoglycan. In the nanomedicine, succinoglycan (RIC) is a type of extracellular polysaccharide secreted by Agrobacterium and rhizobium, belongs to a natural active polysaccharide, has a certain anti-tumor effect, and RIC can exert anti-tumor activity by improving the body's immune function. Resveratrol (RES) is a natural monomer compound that exists in a variety of plants. Resveratrol can exert anti-tumor activity by inhibiting the proliferation of tumor cells, inducing tumor cell apoptosis, inhibiting tumor cell invasion and metastasis, regulating the body's immune function, and directly acting on related targets. The present invention uses succinoglycan as a carrier of nanomedicine, and uses succinoglycan-tetravalent platinum conjugate to encapsulate the anticancer drug small molecule RES, thereby improving the anti-cancer efficacy of the drug, while overcoming the toxic and side effects of platinum drugs and the drug resistance of tumor cells. The determination of the IC50 of cells shows that resveratrol@succinoglycan-tetravalent platinum nanomedicine has a high inhibitory effect on cancer cells. The combined action of various substances in the nanomedicine can significantly increase the toxicity of platinum drugs to HepG2 and LO2 cells, providing a new idea for the construction and application of platinum nanomedicines. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0023] Figure 1 This is a roadmap for constructing the RES@RIC-Pt(IV) nanomedicine in the present invention;

[0024] Figure 2 The UV / visible absorption spectra of RIC, Pt(IV) obtained in Example 1, and RIC-Pt(IV) conjugate;

[0025] Figure 3 The infrared spectra of cisplatin (CDDP), cisplatin oxide (CDDP-OH), tetravalent platinum-succinic anhydride complex (CDDP-OH-SA / Pt(IV)) obtained in Example 1 and RIC-Pt(IV) conjugate;

[0026] Figure 4 1 is the hydrogen nuclear magnetic resonance spectrum of CDDP-OH and Pt(IV) obtained in Example 1;

[0027] Figure 5 The H NMR spectra of RIC, Pt(IV) obtained in Example 1 and RIC-Pt(IV) conjugate;

[0028] Figure 6 Thermogravimetric analysis diagrams of RIC, Pt(IV) obtained in Example 1, and RIC-Pt(IV) conjugate;

[0029] Figure 7 It is the curve diagram of the standard solution of metal platinum;

[0030] Figure 8 is the standard curve of resveratrol;

[0031] Fig. 9 is the SEM image of RIC;

[0032] Fig.10 This is the SEM image of RIC-Pt(IV) obtained in Example 6;

[0033] Fig.11 This is the SEM image of RES@RIC-Pt(IV) obtained in Example 6;

[0034] Fig.12 TEM image of RIC;

[0035] Fig.13 TEM image of RIC-Pt(IV) obtained in Example 1;

[0036] Fig.14 TEM image of RES@RIC-Pt(IV) obtained in Example 1;

[0037] Fig.15 This is the cell survival rate data of HepG2 and LO2 cells after 48 hours;

[0038] Fig.16 IC values ​​of RES, CDDP, RIC-Pt(IV) obtained in Examples 1 and 6, RES1@RIC-Pt(IV), and RES2@RIC-Pt(IV) for HepG2 cells are shown in Table 1. 50 Value data graph;

[0039] Fig.17 IC values ​​of RES, CDDP, RIC-Pt(IV) obtained in Examples 1 and 6, and RES@RIC-Pt(IV) for LO2 cells 50 Value data graph. DETAILED DESCRIPTION

[0040] The present invention provides a resveratrol@succinoglycan-tetravalent platinum nanomedicine, comprising resveratrol and a succinoglycan-tetravalent platinum conjugate layer encapsulating the resveratrol;

[0041] The particle size of the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 500-800 nm;

[0042] The components of the succinoglycan-tetravalent platinum conjugate layer are a conjugate of a tetravalent platinum organic dianhydride complex and succinoglycan.

[0043] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0044] In the present invention, the particle size of the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 500-800 nm, and in a specific embodiment, it can be 600 nm or 700 nm.

[0045] In the present invention, the mass percentage of resveratrol in the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 2-80%, and in a specific embodiment, it can be 3.3% or 20.95%.

[0046] In the present invention, the mass percentage of platinum element in the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 2-20%, and in a specific embodiment, it can be 5% or 10%.

[0047] In the present invention, the components of the succinopolysaccharide-tetravalent platinum conjugate layer are a conjugate of a tetravalent platinum organic dianhydride complex and succinopolysaccharide; and the tetravalent platinum organic dianhydride complex is a tetravalent platinum-succinic anhydride complex.

[0048] In the present invention, the preparation method of the tetravalent platinum-succinic anhydride complex comprises the following steps:

[0049] Cisplatin and hydrogen peroxide solution are mixed and axially oxidized to obtain cisplatin oxide;

[0050] In the present invention, the cisplatin is a cisplatin suspension; the solvent of the cisplatin suspension is water, and the concentration is 0.01-0.05 g / mL, and in a specific embodiment, it can be 0.02 g / mL, 0.033 g / mL or 0.04 g / mL; the concentration of the hydrogen peroxide solution is 10-30%, and in a specific embodiment, it can be 20% or 30%; the volume ratio of the cisplatin suspension to the hydrogen peroxide solution is 1-4:5, and in a specific embodiment, it can be 3:5; the mixing can be dropwise adding the hydrogen peroxide solution to the cisplatin suspension; The temperature of the axial oxidation is 25 to 80°C, and in a specific embodiment, it can be 40°C or 60°C; the reaction time is 2 to 6 hours, and in a specific embodiment, it can be 4 hours; the axial oxidation also includes cooling, a first solid-liquid separation, washing, a second solid-liquid separation and drying in sequence; the cooling temperature is room temperature, and the time is 8 to 12 hours; the first solid-liquid separation is filtration, and the washing reagent is ice water; the second solid-liquid separation is centrifugation; the drying is vacuum drying, and the temperature is 50 to 70°C, and in a specific embodiment, it can be 60°C.

[0051] After the cisplatin oxide is obtained, the cisplatin oxide, succinic anhydride and an organic solvent are mixed to carry out a coordination reaction to obtain the tetravalent platinum-succinic anhydride complex.

[0052] In the present invention, the molar ratio of cisplatin oxide to succinic anhydride is 1:0.2-1, and in a specific embodiment, it can be 1:1; the organic solvent is dimethyl sulfoxide; the mass ratio of cisplatin oxide to the volume ratio of the organic solvent is 0.1-0.6 g:16 mL, and in a specific embodiment, it can be 0.2 g:16 mL; the mixing is: adding succinic anhydride to the organic solvent suspension of cisplatin oxide; the temperature of the coordination reaction is 25-80°C, and in a specific embodiment, it can be 40°C or 60°C; the reaction time is 10-36h, and in a specific embodiment, it can be 24h; after the coordination reaction, it also includes a first drying, recrystallization and a second drying carried out in sequence; the first drying is vacuum drying, and the temperature is 50-70°C, and in a specific embodiment, it can be 60°C; the recrystallization is: dispersed in methanol and precipitated in ice ether; the second drying is vacuum drying, and the temperature is 40-70°C, and in a specific embodiment, it can be 55°C.

[0053] The present invention also provides a method for preparing the resveratrol@succinoglycan-quadrivalent platinum nanomedicine described in the above technical solution, comprising the following steps:

[0054] The tetravalent platinum organic dianhydride complex, succinoglycan, a coupling catalyst and an organic-water mixed solvent are mixed to carry out an esterification reaction to obtain a succinoglycan-tetravalent platinum coupling product;

[0055] Mixing the succinoglycan-quadrivalent platinum conjugate, resveratrol and an organic solvent to obtain a drug-containing organic phase;

[0056] The drug-containing organic phase and water are mixed to obtain the resveratrol@succinoglycan-tetravalent platinum nanomedicine.

[0057] The invention mixes a tetravalent platinum organic dianhydride complex, succinopolysaccharide, a coupling catalyst and an organic-water mixed solvent, and performs an esterification reaction to obtain a succinopolysaccharide-tetravalent platinum coupling compound.

[0058] In the present invention, the mass ratio of the tetravalent platinum organic dianhydride complex to succinoglycan is 217:50-200, and in a specific embodiment, it can be 217:150.

[0059] In the present invention, the coupling catalyst includes 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; the molar ratio of the 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:5-12, and in a specific embodiment, it can be 1:10; the molar ratio of the tetravalent platinum complex and the catalyst is 1:5-15, and in a specific embodiment, it can be 1:11.

[0060] In the present invention, the organic solvent in the organic-water mixed solvent is dimethyl sulfoxide; the volume ratio of the organic solvent to water is 2:0.5-1, and in a specific embodiment, it can be 2:0.8 or 2:1; the dosage ratio of the tetravalent platinum complex and the organic-water mixed solvent is 0.5-2mmol:30mL, and in a specific embodiment, it can be 1mmol:30mL.

[0061] In the present invention, the temperature of the esterification reaction is 15-50° C., and in a specific embodiment, it can be 25° C.; the time is 10-36 hours, and in a specific embodiment, it can be 24 hours; the esterification reaction is carried out under stirring conditions.

[0062] In the present invention, the esterification reaction further includes filtration and freeze-drying performed in sequence; the filtration is dialysis; the molecular weight cutoff of the dialysis is 2000-8000, and in a specific embodiment, it can be 3500.

[0063] After obtaining the succinoglycan-tetravalent platinum conjugate, the present invention mixes the succinoglycan-tetravalent platinum conjugate, resveratrol and an organic solvent to obtain a drug-containing organic phase.

[0064] In the present invention, the mass ratio of the succinoglycan-quadrivalent platinum conjugate to resveratrol is 10:1-8, and in a specific embodiment, it can be 10:5.

[0065] In the present invention, the organic solvent is dimethyl sulfoxide; the dosage ratio of the succinoglycan-quadrivalent platinum conjugate to the organic solvent is 1 g:50 mL.

[0066] In the present invention, the mixing is stirring; the stirring temperature is room temperature, and the stirring time is 10 to 60 minutes, and in a specific embodiment, it can be 30 minutes.

[0067] After obtaining the drug-containing organic phase, the present invention mixes the drug-containing organic phase with water to obtain the resveratrol@succinoglycan-tetravalent platinum nanomedicine.

[0068] In the present invention, the volume ratio of the drug-containing organic phase to water is 1:10-15, and in a specific embodiment, it can be 1:12.

[0069] In the present invention, the mixing is: dropping the drug-containing organic phase into water and stirring; the dropping rate is 1 mL / 2 min to 8 min, and in a specific embodiment, it can be 1 mL / 6 min; the stirring time is 0.5 to 3 h, and in a specific embodiment, it can be 2 h. In the present invention, the self-assembly ability of succinoglycan in aqueous solution is used to encapsulate resveratrol to form a nearly spherical morphological structure.

[0070] In the present invention, the mixing further includes filtering and freeze-drying performed in sequence; the filtration includes dialysis and injection filter filtration performed in sequence; the molecular weight cutoff of the dialysis is 2000-8000, and in a specific embodiment, it can be 3500; the dialysis reagent is distilled water; the dialysis time is 12-72h, and in a specific embodiment, it can be 36h; the pore size of the injection filter is 0.22-0.45μm, and in a specific embodiment, it can be 0.45μm.

[0071] In the preparation method provided by the present invention, the RIC-Pt (IV) conjugate can be prepared under mild experimental conditions, and the natural polysaccharide nano drug system encapsulating the small molecule RES of the anticancer drug is prepared by adjusting the dosage of RES, and the process is simple and convenient. Experiments have found that the effect of the RES@RIC-Pt (IV) nano drug system on HepG2 and LO2 is concentration-dependent to a certain extent. As the concentration of the drug increases, the effect of the drug on the cells becomes greater, and the combined action of the nano drugs can significantly increase the toxicity of the platinum drug to HepG2 and LO2 cells, which provides a new idea for using natural polysaccharides as drug carrier materials, assembling resveratrol and platinum drugs, and constructing a combined drug system.

[0072] The present invention also provides the use of the resveratrol@succinoglycan-tetravalent platinum nanomedicine described in the above technical solution or the resveratrol@succinoglycan-tetravalent platinum nanomedicine obtained by the preparation method described in the above technical solution in the preparation of anti-tumor preparations.

[0073] In the present invention, the anti-tumor preparation can be an anti-HepG2 cell preparation or an anti-LO2 cell preparation; the present invention does not have any special limitation on the application process of the resveratrol@succinoglycan-tetravalent platinum nanomedicine in the preparation of anti-tumor preparations, and the method familiar to those skilled in the art can be used.

[0074] In order to further illustrate the present invention, the resveratrol@succinoglycan-quadrivalent platinum nanomedicine provided by the present invention and its preparation method and application are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0075] The main reagents and instruments used in the embodiments and test examples of the present invention are shown in Tables 1 and 2.

[0076] Table 1 Main reagents for the experiments of the embodiments and test examples of the present invention

[0077]

[0078] Table 2 Main instruments and equipment for the embodiments and test examples of the present invention

[0079]

[0080]

[0081] The present invention provides a method for preparing resveratrol@succinoglycan-quadrivalent platinum nanomedicine. The construction route of the nanomedicine is as follows: Figure 1 The specific steps are shown in the following embodiments.

[0082] Example 1

[0083] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0084] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 25°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0085] 50 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0086] Example 2

[0087] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 50°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0088] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 25°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0089] 50 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0090] Example 3

[0091] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0092] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 35°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0093] 50 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0094] Example 4

[0095] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0096] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 45°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0097] 50 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0098] Example 5

[0099] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0100] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted at 55°C with magnetic stirring for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0101] 50 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0102] Example 6

[0103] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0104] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 25°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0105] 10 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0106] Example 7

[0107] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0108] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 25°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0109] 20 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0110] Example 8

[0111] Hydrogen peroxide (30%, 20mL) was added dropwise to a suspension of cisplatin (0.4g, 1.33mmol) in water (12mL) at a rate of 1mL / 6min. After reacting at 60°C for 4h, the bright yellow solution was cooled at room temperature overnight to produce yellow crystals. The crystals were filtered, washed with ice water, and centrifuged in sequence, and then vacuum dried at 60°C. Succinic anhydride (0.06g, 0.6mmol) was added to a suspension of the above sample (0.2g, 0.6mmol) in DMSO (16mL), and stirred at 60°C for 24h to obtain a bright yellow solution. The solution was dried at 60°C in a vacuum drying oven, then dispersed with 3mL of methanol, precipitated into 15mL of ice ether, and allowed to stand for 5 minutes to obtain a light yellow solid, which was vacuum dried at 55°C to obtain a tetravalent platinum-succinic anhydride complex (Pt(IV)).

[0112] Succinoglycan (150 mg) and Pt(IV) (217 mg, 0.5 mmol) were added to a round-bottom flask, DMAP (61 mg, 0.5 mmol) and EDC·HCl (958.5 mg, 5 mmol) were added to a mixed solution of DMSO (10 mL) and pure water (5 mL), added to the flask, and the four drugs were completely dissolved by ultrasound, and then reacted with magnetic stirring at 25°C for 24 hours. Then, the reaction solution was dialyzed (MWCO=3500) for 72 hours to remove DMSO and unreacted Pt(IV) complex, and finally freeze-dried at -55°C to obtain RIC-Pt(IV) conjugate.

[0113] 30 mg of RES and RIC-Pt(IV) conjugate (100 mg) were dissolved in DMSO (5 mL) and stirred at room temperature for 30 min. The mixed solution was added dropwise to deionized water (60 mL) within 30 min and continued to stir for 2 h. The mixed solution was then placed in a dialysis bag (MWCO = 3500) and dialyzed with distilled water for 36 h to remove DMSO. The unloaded RES was filtered with a 0.45 μm injection filter, and the resulting clear solution was freeze-dried at -55 °C to obtain RES@RIC-Pt(IV) nanomedicine.

[0114] Test Example 1

[0115] UV / Visible Spectrophotometer (UV-Vis) Testing

[0116] RIC, Pt(IV) obtained in Example 1 and RIC-Pt(IV) nanoparticles were dissolved in deionized water to prepare a sample solution with a mass concentration of 0.5 mg / mL. The samples were tested using a LAMBDA25 UV / Vis spectrophotometer. The wavelength range was set to scan at 200-500 nm, and 4 mL of reference solvent pure water was accurately transferred into two quartz cuvettes. Then, the sample was automatically zeroed. Then, one of the quartz cuvettes was taken out and the test solution corresponding to the reference solvent was added thereto for measurement. The test results are shown in Figure 2 .

[0117] Depend on Figure 2 It can be seen that the UV / visible absorption peaks of RIC, Pt(IV) and RIC-Pt(IV) are located between 250-300nm. The absorption peak of RIC polysaccharide aqueous solution is weak; the absorption peak of Pt(IV) DMSO solution is located near 260nm, and the peak is strong; the absorption peak of RIC-Pt(IV) aqueous solution is located near 275nm, which is red-shifted compared with the absorption peak of Pt(IV) solution, proving that substituents are introduced and the polarity of the solution changes due to the occurrence of esterification reaction.

[0118] Test Example 2

[0119] Infrared spectroscopy (FT-IR) test

[0120] The infrared spectra of RIC, Pt(IV) obtained in Example 1 and RIC-Pt(IV) samples were tested by potassium bromide tableting method. The test method is as follows: the dried sample is placed in an agate mortar, 100-200 times the mass of the sample is added with KBr, ground into fine powder, dried for 5 minutes, and pressed into thin slices in a mold after cooling. The sample is tested by Fourier transform infrared-Raman spectrometer, and the test wave number range is 4000-500cm -1 The test results are shown in Figure 3 .

[0121] Depend on Figure 3 It can be seen that the cis- and trans-[Pt(NH3)2Cl2(OH)2] obtained by oxidation of cisplatin are at 3464 cm -1 There is a strong -OH stretching vibration peak at 1075cm -1 There is a strong Pt-OH bending vibration peak at 542 cm -1 The moderately strong stretching vibration peak of Pt-N(O) at 542cm -1 The weakened absorption peak at 1636 cm -1 The strong peak of -PtOOC-C- and 1715cm -1 The moderate intensity peak of CO-OH at 3420 cm-1 clearly indicated the successful synthesis of CDDP-OH-SA; the peak of succinoglycan at 3420 cm-1 clearly indicated the successful synthesis of CDDP-OH-SA. -1 There is a strong -OH stretching vibration peak at 1700 cm -1 The strong characteristic peak of -COO- at the position proves that the hydroxyl group of RIC and the carboxyl group of Pt(IV) have undergone esterification reaction.

[0122] Test Example 3

[0123] H NMR ( 1 H NMR) test

[0124] Under the conditions of 25°C and 400 MHz, deuterated dimethyl sulfoxide or deuterated water was selected as the solvent, and the CDDP-OH, RIC, Pt(IV) obtained in Example 1 and RIC-Pt(IV) samples were scanned by a nuclear magnetic resonance spectrometer. 1 HNMR spectrum. Test results are shown in Figures 4-5 .

[0125] CDDP-OH and Pt(IV) were dissolved in DMSO-d6 and tested 1 HNMR spectrum Figure 4 .Depend on Figure 4It can be seen from the spectrum of CDDP-OH that the chemical shift near 7.13ppm belongs to the amino group, and the chemical shift at 4ppm-5ppm belongs to the axial hydroxyl group of cisplatin. From the spectrum of Pt(IV), it can be seen that the triplet peak near 5.90ppm is assigned to the NH3 ligand of Pt(IV), the chemical shift at 2.46ppm belongs to the methylene proton, and the ratio of the -NH3 to -CH2- peak areas is equal to the ratio of the corresponding number of hydrogen atoms (3:2), proving the successful synthesis of Pt(IV).

[0126] RIC polysaccharide was dissolved in deuterated water, Pt(IV) and RIC-Pt(IV) conjugate were dissolved in DMSO-d6, and the tested 1 HNMR spectrum Figure 5 .Depend on Figure 5 It can be seen that the chemical shift of 1-1.5ppm belongs to the methylene group in the RIC polysaccharide structure, and the chemical shift of 6.2-7ppm belongs to the NH3 ligand of Pt(IV). The chemical shift has changed and the ratio of the -NH3 to -CH2- peak areas is approximately equal to the ratio of the corresponding number of hydrogen atoms (3:2), which proves the synthesis of RIC-Pt(IV) conjugate.

[0127] Test Example 4

[0128] Thermogravimetric (TGA) testing

[0129] The thermal stability of polysaccharides and tetravalent platinum was studied using a thermogravimetric analyzer. Accurately weigh 4 to 8 mg of RIC, Pt(IV) obtained in Example 1, and RIC-Pt(IV) samples, put them into an alumina crucible and cover the crucible with a cap, and test them on a thermogravimetric analyzer, setting the gas atmosphere to nitrogen, the temperature range to 25 to 800°C, and the heating rate to 10°C / min. The test results are shown in Figure 6 .

[0130] Depend on Figure 6It can be seen that Pt(IV) has little mass loss in the stage of 25-200℃, which may be caused by the weight loss of bound water in the Pt(IV) sample; in the range of 210-300℃, the mass loss rate is the fastest, which may be due to the heat loss caused by the carboxyl or hydroxyl groups or depolymerization in the Pt(IV) sample; at 300-800℃, the mass loss rate slows down and finally stabilizes, and only metal platinum should be left in the sample. RIC polysaccharide has less heat loss in the stage of 25-130℃, which may be caused by the weight loss of bound water in the polysaccharide sample; at 260-340℃, the heat loss amplitude is the largest, which is caused by the thermal decomposition weight loss caused by the dehydration or depolymerization of the sugar ring of the polysaccharide sample itself. Compared with RIC polysaccharide, RIC-Pt(IV) sample has less heat loss in the stage of 25-120℃, which may be caused by the volatilization of bound water in the sample; at 260-320℃, the heat loss amplitude is faster, which is due to the decomposition of some carbon structures of the polysaccharide sugar ring in the conjugate. When Pt(IV) is modified onto the carbon skeleton of RIC, the heat loss rate in the high temperature stage (300-800°C) slows down, the spatial structure becomes more stable, and the thermal stability is improved.

[0131] Test Example 5

[0132] Determination of platinum content by inductively coupled plasma optical emission spectrometry (ICP-OES)

[0133] The 100 mg / L platinum standard solution was diluted with 10% hydrochloric acid to obtain platinum standard solutions with concentrations of 2 mg / L, 4 mg / L, 6 mg / L, 8 mg / L, and 10 mg / L, respectively. The standard curve of platinum was determined by ICP-OES. See the standard curve for details. Figure 7 .

[0134] The RIC-Pt(IV) obtained in Example 1 was weighed on a weighing balance to obtain two 5 mg samples, each of which was moistened with 1 mL of ultrapure water, and 2 mL of aqua regia was added to carry out nitration in an oil bath at 125°C. The process was repeated once or more until the cotton-like sample was basically completely dissolved. The solution was filtered with a 0.45 μm disposable syringe filter, and the clarified solution after filtration was transferred to a 10.00 mL brown volumetric flask. After constant volume, the Pt content in the RIC-Pt(IV) conjugate was determined by ICP-OES.

[0135] according to Figure 7 The metal Pt content in the RIC-Pt(IV) conjugate was determined by inductively coupled plasma emission spectrometry to be 58.41 mg / L and 58.68 mg / L, and the average value of the two was 58.55 mg / L.

[0136] Test Example 6

[0137] Resveratrol content test

[0138] 5 mg of resveratrol was dissolved in a mixed solution of 10 mL of methanol and pure water (4.5 mL of methanol, 5.5 mL of pure water) to obtain a 0.5 mg / mL resveratrol solution, which was then diluted to different concentrations of RES solutions (2, 4, 6, and 8 μg / mL). A standard curve was drawn with the mass concentration of resveratrol as the horizontal axis and the absorbance as the vertical axis. The standard curve is shown in Figure 8 .

[0139] The resveratrol content in the nanomedicine was determined. The method is as follows: 5 mg of RES@RIC-Pt(IV) obtained in Example 1 and Example 6 were respectively dissolved in 10 mL of deionized water, stirred and sonicated to dissolve. Then, the absorbance was measured at 305 nm, and the resveratrol content in the sample was calculated. Example 6 and Example 1 correspond to RES1@RIC-Pt(IV) and RES2@RIC-Pt(IV).

[0140] Substitute the absorbance of the tested RES@RIC-Pt(IV) sample into Figure 8 In the resveratrol standard curve, the resveratrol concentration in the sample was calculated. Through the formula conversion, it was calculated that the resveratrol content in the RES1@RIC-Pt(IV) sample was 3.3%, the drug loading efficiency was 10.25%, and the resveratrol content in the RES2@RIC-Pt(IV) sample was 20.95%, and the drug loading efficiency was 55.71%.

[0141] Test Example 7

[0142] Scanning Electron Microscope (SEM) Testing

[0143] The surface morphology of RIC, Pt(IV) obtained in Example 6 and RIC-Pt(IV) freeze-dried samples was observed using a scanning electron microscope (KYKY-EM6200 / 6900) with an accelerating voltage of 15 kV. The sample solution after the reaction was freeze-dried in a freeze dryer (LGJ-10P) at -55°C for 48 hours to obtain a cotton-like sample. During the test, tweezers were used to clamp the sample and stick it on a conductive tape, and an ear bulb was used to blow away the debris around the sample, and then the microscopic morphology of the sample was observed under a KYKY-EM6200 / 6900 electron microscope. The test results are shown in Figures 9-11 .

[0144] Depend on Figures 9-11 It can be seen that the surface of RIC polysaccharide is relatively smooth, the surface fibers are dense, and a small amount of cotton-like flocs are observed after magnification; after being modified by tetravalent platinum, the fiber structure on the surface of the polysaccharide is destroyed, changing from cotton-like to thin strips with clearer strip boundaries; after wrapping resveratrol, the three-dimensional spatial structure is further destroyed, the structure becomes unstable from entanglement, the surface morphology becomes looser, and the strip boundaries become blurred.

[0145] Test Case 8

[0146] Transmission Electron Microscope (TEM) Test

[0147] The morphology of RIC-Pt(IV) nanospheres and RES@RIC-Pt(IV) vesicles obtained in Example 1 was observed using a JEOL JEM-1400plus transmission electron microscope from Malvern Panalytical, the Netherlands, at an accelerating voltage of 100 kV. A drop of the sample to be tested (0.5 mg / mL) was placed on a 400-mesh carbon-coated copper grid, negatively stained with 2% phosphotungstic acid, and dried at room temperature. The test results are shown in Figures 12-14 , denoted as RES1@RIC-Pt(IV) in the figure.

[0148] Depend on Figures 12-14 It can be seen that the morphology of the RIC-Pt(IV) conjugate in aqueous solution is approximately spherical or ellipsoidal, with a size of 200-500nm, and a small amount of 500-800nm. The TEM image of RES1@RIC-Pt(IV) shows that the sample in aqueous solution is approximately spherical or ellipsoidal, with a size larger than RIC-Pt(IV), mainly 500-800nm. As the amount of resveratrol added increases, the boundaries of the spherical morphology become clearer and the size increases further. The amount of resveratrol encapsulated inside increases, the encapsulation becomes tighter, the internal space structure becomes more three-dimensional, and the drug loading capacity and efficiency are enhanced.

[0149] Test Example 9

[0150] Cell culture and cytotoxic activity assay

[0151] This test uses HepG2 (hepatoblastoma cell line) and LO2 (contaminated Hela-derived cell line) and uses the MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-2,5-diphenyltetrazolium bromide] method for testing. The MTT method is a method for detecting the number of living cells. The specific steps of the MTT cell proliferation and cytotoxicity test are as follows:

[0152] HepG2 and LO2 cells were cultured in DMEM medium supplemented with 10% (v / v) fetal bovine serum. The culture atmosphere was 95% air (containing 5% CO2) and 37°C. 100 μL of cell-containing culture medium (about 6000-8000 cells per well) was added to each well of the 96-well culture plate. After 24 hours of adherent growth, 100 μL of sample solution with a concentration gradient of 0-256 μM was added to each well. The cells were cultured for 48 hours after drug addition, and then 50 μL of MTT solution (0.5 mg / mL) was added to each well. After standing for 4 hours, a blue-purple precipitate was produced. The supernatant was discarded, and 150 μL of DMSO was added to dissolve the precipitate. The absorbance of each well at 570 nm was measured on a multifunctional microplate reader. The percentage of surviving cells was calculated relative to the control group without drug addition, and each group of experiments was measured in parallel three times. The formula for evaluating the efficacy of the sample on cancer cells is as follows:

[0153] Viable cell (%) = (OD sample -OD blank ) / (OD control -OD blank )×100%

[0154] Among them, OD sample is the OD value of the added sample, OD control is the OD value without adding sample, OD blank is the blank OD value without cells.

[0155] Cell viability curves of HepG2 and LO2 after 48 h, and IC values ​​of RES, CDDP, Pt(IV) obtained in Example 6, 1, RIC-Pt(IV), RES1@RIC-Pt(IV), and RES2@RIC-Pt(IV) 50 Value Figures 15 to 17 .

[0156] Depend on Figures 15 to 17 It can be seen that the effects of the six drugs on the two types of cells show a certain degree of concentration dependence. As the drug concentration increases, the effect of the drug on the cells becomes greater. In addition, the combined effect of the drugs RES1@RIC-Pt(IV) and RES2@RIC-Pt(IV) on the cells is greater than that of RES, CDDP, Pt(IV), and RIC-Pt(IV) alone. Therefore, the combined effect of RES1@RIC-Pt(IV) and RES2@RIC-Pt(IV) nanomedicines can significantly increase the toxicity of platinum drugs to HepG2 and LO2 cells.

[0157] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A resveratrol@succinoglycan-quadrivalent platinum nanomedicine, characterized in that: The invention comprises resveratrol and a succinoglycan-quadrivalent platinum conjugate layer encapsulating the resveratrol; The particle size of the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 500-800 nm; The components of the succinoglycan-tetravalent platinum conjugate layer are a conjugate of a tetravalent platinum organic dianhydride complex and succinoglycan.

2. The resveratrol@succinoglycan-quadrivalent platinum nanomedicine according to claim 1, characterized in that: The mass percentage of resveratrol in the resveratrol@succinoglycan-quadrivalent platinum nanomedicine is 2-80%.

3. The resveratrol@succinoglycan-quadrivalent platinum nanomedicine according to claim 1, characterized in that: The mass percentage of platinum element in the resveratrol@succinoglycan-tetravalent platinum nanomedicine is 2-20%.

4. The resveratrol@succinoglycan-quadrivalent platinum nanomedicine according to claim 1, characterized in that: The tetravalent platinum organic dianhydride complex is a tetravalent platinum-succinic anhydride complex.

5. The method for preparing the resveratrol@succinoglycan-quadrivalent platinum nanomedicine according to any one of claims 1 to 4, characterized in that: The following steps are involved: The tetravalent platinum organic dianhydride complex, succinoglycan, a coupling catalyst and an organic-water mixed solvent are mixed to carry out an esterification reaction to obtain a succinoglycan-tetravalent platinum coupling product; Mixing the succinoglycan-quadrivalent platinum conjugate, resveratrol and an organic solvent to obtain a drug-containing organic phase; The drug-containing organic phase and water are mixed to obtain the resveratrol@succinoglycan-tetravalent platinum nanomedicine.

6. The preparation method according to claim 5, characterized in that: The mass ratio of the tetravalent platinum organic dianhydride complex to succinopolysaccharide is 1:1-5.

7. The preparation method according to claim 5, characterized in that: The coupling catalyst includes 4-dimethylaminopyridine and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

8. The preparation method according to claim 5, characterized in that: The temperature of the esterification reaction is 15-50° C., and the reaction time is 10-36 hours.

9. The preparation method according to claim 5, characterized in that: The mass ratio of the succinoglycan-quadrivalent platinum conjugate to resveratrol is 10:1-8.

10. Use of the resveratrol@succinoglycan-tetravalent platinum nanodrug according to any one of claims 1 to 4 or the resveratrol@succinoglycan-tetravalent platinum nanodrug obtained by the preparation method according to any one of claims 5 to 9 in the preparation of anti-tumor preparations.

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