Nanoformations of cisplatin-based anticancer drugs and preparation and application of nanoformations that enhance the efficacy of cisplatin-based anticancer drugs

Abstract

This invention discloses a nano-formulation of a cisplatin-based anticancer drug and the preparation and application of nano-formulations that enhance the efficacy of cisplatin-based anticancer drugs. The preparation includes: dissolving polyglutamic acid and cisplatin in deionized water and stirring to obtain a cisplatin-polyglutamic acid conjugate solution; adding a surfactant solution with opposite charge to obtain a cisplatin-polyglutamic acid nanoparticle dispersion; centrifuging, washing with water, and drying the product to obtain cisplatin-polyglutamic acid nanoparticles. Centrifuging and redispersing the cisplatin-polyglutamic acid nanoparticle dispersion, adding potassium permanganate, stirring, centrifuging, washing with water, and drying to obtain a MnO2-cisplatin-polyglutamic acid nano-formulation, which is the nano-formulation for enhancing the efficacy of cisplatin-based anticancer drugs. The nanoparticles prepared by this invention have a solid spherical structure. This method for preparing platinum-based anticancer drug nano-formulations is efficient, has a high yield, and is low in cost. It can be used to prepare drugs for treating tumors and has great application potential in the fields of drug delivery, controlled release, and tumor treatment.

Description

Technical Field

[0001] This invention pertains to the preparation of pharmaceutical nanoparticles, specifically relating to a nanoparticle formulation of a cisplatin-based anticancer drug and the preparation and application of nanoparticle formulations that enhance the efficacy of cisplatin-based anticancer drugs. Background Technology

[0002] With the development of nanotechnology, combining nanotechnology with pharmaceuticals can yield nanomedicines with specific dosage forms, namely drug-loaded particles or nanomedicine crystals with particle sizes of 1-1000 nm, made from drugs and excipients. During the preparation process, the drug can achieve a stable, dispersed nanoscale state through the action of a small amount of surfactants or polymeric excipients. Drug nanoformulations play a crucial role in improving drug utilization, enhancing safety, and improving therapeutic efficacy. Especially for some therapeutic drugs, such as anticancer drugs, which are very expensive, have strong toxic side effects, are difficult to store and transport, and require repeated administration, nanomedicine formulations offer hope for treatment.

[0003] As one of the most representative platinum-based anticancer drugs in clinical practice, cisplatin and its derivatives have shown certain killing effects on various solid tumors, including ovarian cancer, testicular cancer, lung cancer, esophageal cancer, malignant lymphoma, head and neck squamous cell carcinoma, and thyroid cancer. As one of the most widely used platinum-based anticancer drugs, cisplatin and its derivatives possess strong broad-spectrum anticancer activity, primarily exerting their antitumor effects by damaging tumor cell DNA, inhibiting replication, and inducing apoptosis. However, it has significant toxic side effects, easily causing hepatotoxicity, nephrotoxicity, cardiotoxicity, neurotoxicity, and hematologic toxicity, which greatly limits its further clinical application. Furthermore, the efficacy of platinum-based anticancer drugs is greatly influenced by the tumor microenvironment. The tumor microenvironment has characteristics different from those of normal cells, typically composed of high concentrations of glutathione (GSH) and hydrogen peroxide (H2O2), weak acidity, and hypoxia. In particular, the concentration of GSH in tumor cells is extremely high; however, high concentrations of GSH can bind with Pt to form Pt(GS)2 conjugates, which are rapidly exported from tumor cells. Therefore, the increase in thiol substances leads to cytoplasmic detoxification of platinum-based anticancer drugs in tumor cells, thereby reducing the antitumor sensitivity and efficacy of platinum-based anticancer drugs. In addition, after cisplatin enters the bloodstream, it is bound to thiol-containing proteins such as human serum albumin, thus rendering it ineffective.

[0004] Polyglutamic acid (γ-PGA) is a highly water-soluble, biodegradable, and non-toxic biopolymer. γ-PGA is a unique anionic natural polymer, a homopolymer composed of α-amine and γ-carboxyl groups linked by amide bonds. γ-PGA possesses excellent water solubility, strong adsorption capacity, and biodegradability, with its degradation product being non-toxic glutamic acid. In recent years, increased environmental awareness and the requirements of national sustainable development strategies have driven the research and development of polyglutamic acid industrialization. The molecular weight of γ-PGA and its derivatives ranges from 1000 kDa to 2000 kDa, thus γ-PGA can be widely used in the food industry, cosmetics, drug delivery, medical applications, and hydrogels. Summary of the Invention

[0005] Purpose of the invention: To address the problems existing in the prior art, this invention provides a method for preparing nano-formulations of platinum-based anticancer drugs, represented by cisplatin. The nano-formulations prepared by this invention can effectively solve the problems of strong toxic side effects and the need for repeated administration of cisplatin-based chemotherapy drugs.

[0006] Another objective of this invention is to provide a method for preparing a nano-formulation that enhances the efficacy of cisplatin-based anticancer drugs. The nano-formulation prepared by this invention can be used to improve chemotherapy sensitivity and efficacy.

[0007] The present invention also provides nano-formulations of the aforementioned cisplatin-based anticancer drugs, nano-formulations that enhance the efficacy of cisplatin-based anticancer drugs, and their applications.

[0008] Technical solution: To achieve the above-mentioned objective, the present invention provides a method for preparing a nano-formulation of a cisplatin-based anticancer drug, comprising the following steps:

[0009] (1) Preparation of cisplatin-polyglutamic acid conjugate: Polyglutamic acid and cisplatin were dissolved in deionized water and stirred under light-protected conditions to obtain a cisplatin-polyglutamic acid conjugate dispersion;

[0010] (2) Preparation of cisplatin-polyglutamic acid nanoparticles: Under continuous stirring, a surfactant solution with opposite charge was added to the cisplatin-polyglutamic acid conjugate solution obtained in step (1), and stirring was continued to obtain a cisplatin-polyglutamic acid nanoparticle dispersion.

[0011] (3) Centrifuge to collect the solid precipitate in the dispersion of step (2), then centrifuge, wash with water and dry the product to obtain cisplatin-polyglutamic acid nanoparticles.

[0012] In step (1), the relative molecular mass of polyglutamic acid is 1000 Da-700 kDa.

[0013] In step (1), the concentration ratio of cisplatin to polyglutamic acid is 0.001-100:1.

[0014] In step (2), the surfactant is selected from surfactants with opposite charge to the product of step (1), such as surfactants with positive charge such as quaternary ammonium salts, including octadecyltrimethylammonium bromide or hexadecyltrimethylammonium bromide CTAB, and the concentration of the surfactant is 0.0001 g / mL to 0.1 g / mL.

[0015] Preferably, the surfactant is hexadecyltrimethylammonium bromide (CTAB).

[0016] In step (2), the volume ratio of the cisplatin-polyglutamic acid conjugate dispersion to the surfactant solution is 1-1000:1.

[0017] Preferably, the volume ratio of the cisplatin-polyglutamic acid conjugate solution to the CTAB solution is 20-40:1.

[0018] In steps (1) and (2), the stirring rate is 0-10000 r / min and the reaction temperature is 0-100℃.

[0019] Preferably, in steps (1) and (2), the stirring rate is 0-1500 r / min, and the preferred temperature is 20-40℃.

[0020] Further, step (1) is as follows: 250 mg of polyglutamic acid with a relative molecular mass of 1000 Da is dissolved in 36 mL of deionized water, 140 mg of cisplatin is added to the polyglutamic acid solution, and the mixture is stirred continuously at 1500 r / min for 24 h under light-protected conditions; 1.5 mL of CTAB solution with a concentration of 0.03645 g / mL is quickly added to the obtained cisplatin-polyglutamic acid conjugate solution, and the mixture is stirred continuously at 1500 r / min. The obtained nanoparticles are centrifuged, washed with water, and dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0021] In step (2), the drying method is freeze drying or vacuum drying.

[0022] The present invention relates to a method for preparing nano-formulations that enhance the efficacy of cisplatin-based anticancer drugs, comprising the following steps:

[0023] The prepared cisplatin-polyglutamic acid nanoparticles were centrifuged and redispersed, then potassium permanganate was added, and after stirring, the product was centrifuged, washed with water, and dried to obtain MnO2-cisplatin-polyglutamic acid nano-formulation, which is a nano-formulation to enhance the efficacy of cisplatin-based anticancer drugs.

[0024] Preferably, the mass ratio of polyglutamic acid to potassium permanganate in step (1) is 1-100:1.

[0025] Preferably, freeze drying or vacuum drying is used to obtain the MnO2-cisplatin-polyglutamic acid nanoparticle formulation.

[0026] The present invention relates to a method for preparing nano-formulations of cisplatin-based anticancer drugs, which yields nano-formulations of cisplatin-based anticancer drugs.

[0027] The nano-formulation of the cisplatin-based anticancer drug has a solid spherical structure, and the particle size of the nanoparticles is 2nm-10μm.

[0028] Preferably, the particle size of the cisplatin-polyglutamic acid nanoparticles is 10nm-200nm, and most preferably 50nm-150nm.

[0029] The present invention relates to a method for preparing nano-formulations that enhance the efficacy of cisplatin-based anticancer drugs, which produces nano-formulations that enhance the efficacy of cisplatin-based anticancer drugs.

[0030] The MnO2-cisplatin-polyglutamic acid nanoparticles have a particle size of 2nm-10μm and a MnO2 shell thickness of 0-5μm.

[0031] Preferably, the particle size of the MnO2-cisplatin-polyglutamic acid nanoparticles is 10nm-200nm, most preferably 50nm-150nm, and the thickness of the MnO2 shell is most preferably 0.01nm-50nm.

[0032] The application of the nano-formulation of cisplatin-based anticancer drugs or the nano-formulation that enhances the efficacy of cisplatin-based anticancer drugs described in this invention in the preparation of drugs for treating tumors.

[0033] The nanoparticles prepared by this invention have a solid spherical structure, and the preparation method of this platinum-based anticancer drug nanoparticle formulation is efficient, has a high yield, and is low in cost. This invention first uses polymers such as polyglutamic acid, which can react with chlorine and / or amino functional groups, as a preliminary carrier; then, utilizing the interaction between the chlorine and / or amino functional groups of platinum-based anticancer drugs such as cisplatin and the functional groups of the polymer molecular chain, cisplatin and other platinum-based anticancer drugs are loaded onto polymer molecular chains such as polyglutamic acid; finally, under the action of a surfactant with an opposite charge to the cisplatin-polymer conjugate, the drug-loaded polymer chains spontaneously aggregate into nanoparticles; these nanoparticles have enormous application potential in drug delivery, controlled release, and tumor therapy.

[0034] As a redox reactive substance, manganese dioxide (MnO2) can react with excess GSH and other substances in tumor cells. The reaction mechanism is as follows (1-3): 2GSH + MnO2 + 2H + →GSSG+Mn 2++2H2O(1); MnO2+2H + →Mn 2+ +H2O+1 / 2O2(2);MnO2+H2O2+2H + →Mn 2+ +2H2O+O2(3); Based on these principles, MnO2 is widely used in photodynamic therapy (PDT), magnetic resonance imaging (MRI), etc. The redox reaction between MnO2 and GSH in tumor cells is used to consume GSH to improve the chemosensitivity and efficacy of platinum-based anticancer drugs. This invention utilizes MnO2 to consume GSH to improve the chemosensitivity and efficacy of platinum-based anticancer drugs, and prepares a nano-formulation that enhances the efficacy of cisplatin-based anticancer drugs.

[0035] This invention designs two types of nanoparticles: nano-formulations of cisplatin-based anticancer drugs and nano-formulations that enhance the efficacy of cisplatin-based anticancer drugs. The principle behind the cisplatin-based anticancer drug nano-formulation is as follows: First, a cisplatin-polyglutamic acid conjugate carrying cisplatin is obtained through the coordination of polyglutamic acid and cisplatin. However, the cisplatin in this conjugate remains exposed and will still bind to thiol-containing human serum albumin and other substances upon contact with blood, thus rendering it ineffective. Therefore, it is necessary to isolate cisplatin from plasma. Thus, a cisplatin-based anticancer drug nano-formulation is prepared using a surfactant with an opposite charge to the cisplatin-polyglutamic acid conjugate, based on the principle of electrostatic self-assembly. The nano-formulation that enhances the efficacy of cisplatin-based anticancer drugs is designed to address the phenomenon where cisplatin and other platinum-based anticancer drugs, or the cisplatin-polyglutamic acid conjugate, bind to glutathione and become ineffective after entering tumor cells. The principle utilizes a MnO2 shell to first consume glutathione within tumor cells through a redox reaction, thereby achieving the enhanced drug effect. Polyglutamic acid is a biodegradable material that can be degraded in the body and has no toxic side effects.

[0036] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0037] (1) The materials used in the preparation of cisplatin-polyglutamic acid nanoparticles and MnO2-cisplatin-polyglutamic acid nanoparticles by the present invention are biodegradable materials with good biocompatibility.

[0038] (2) The cisplatin-polyglutamic acid nanoparticles and MnO2-cisplatin-polyglutamic acid nano-formulations described in this invention are nano to micron scale, and have excellent scale advantages.

[0039] (3) The preparation methods of cisplatin-polyglutamic acid nanoparticles and MnO2-cisplatin-polyglutamic acid nano-formulation prepared by the present invention are rapid and simple, and have industrialization and practical application value.

[0040] (4) The cisplatin-polyglutamic acid nanoparticles and MnO2-cisplatin-polyglutamic acid nano-formulations prepared by this invention are not only rich and readily available in raw materials, but also have high yield and low preparation cost, and have strong prospects for industrial production.

[0041] (5) The cisplatin-polyglutamic acid nanoparticles and MnO2-cisplatin-polyglutamic acid nano-preparations prepared by the present invention have stable physicochemical properties and can be stored and transported for a long time.

[0042] (6) The MnO2-cisplatin-polyglutamic acid nano-formulation prepared in this invention improves the killing effect on tumor cells by 2-3 times. Attached Figure Description

[0043] Figure 1 This is a transmission electron microscope image of the cisplatin-polyglutamic acid conjugate prepared in Example 1;

[0044] Figure 2 Transmission electron microscope image of cisplatin-polyglutamic acid nanoparticles prepared in Example 8;

[0045] Figure 3 The UV spectrum of the cisplatin-polyglutamic acid nanoparticles prepared in Example 12 is shown below.

[0046] Figure 4 Fourier transform infrared spectrum of cisplatin-polyglutamic acid nanoparticles prepared in Example 14;

[0047] Figure 5 The UV spectrum of the MnO2-cisplatin-polyglutamic acid nanoparticles prepared in Example 18;

[0048] Figure 6 This is a transmission electron microscope image of the MnO2-cisplatin-polyglutamic acid nanoformulation prepared in Example 18.

[0049] Figure 7 The image shows the cell apoptosis detection results of cisplatin and MnO2-cisplatin-polyglutamic acid nanoparticles obtained in Example 22. Detailed Implementation

[0050] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0051] Unless otherwise specified, all materials and reagents used in the embodiments are commercially available.

[0052] Example 1

[0053] Preparation of cisplatin-polyglutamic acid conjugate

[0054] 250 mg of polyglutamic acid with a relative molecular mass of 1000 Da was dissolved in 36 mL of deionized water. 140 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 1500 r / min for 24 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0055] Transmission electron microscope image of the cisplatin-polyglutamic acid conjugate prepared in Example 1 is shown below. Figure 1 As shown.

[0056] Example 2

[0057] Preparation of cisplatin-polyglutamic acid conjugate

[0058] 250 mg of polyglutamic acid with a relative molecular mass of 70 kDa was dissolved in 36 mL of deionized water. 140 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 1500 r / min for 24 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0059] Example 3

[0060] Preparation of cisplatin-polyglutamic acid conjugate

[0061] 250 mg of polyglutamic acid with a relative molecular mass of 2000 Da was dissolved in 36 mL of deionized water. 140 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 1500 r / min for 24 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0062] Example 4

[0063] Preparation of cisplatin-polyglutamic acid conjugate

[0064] 250 mg of polyglutamic acid with a relative molecular mass of 1000 Da was dissolved in 36 mL of deionized water. 140 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 750 r / min for 24 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0065] Example 5

[0066] Preparation of cisplatin-polyglutamic acid conjugate

[0067] 250 mg of polyglutamic acid with a relative molecular mass of 2000 Da was dissolved in 36 mL of deionized water. 140 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 100 r / min for 24 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0068] Example 6

[0069] Preparation of cisplatin-polyglutamic acid conjugate

[0070] 250 mg of polyglutamic acid with a relative molecular mass of 2000 Da was dissolved in 36 mL of deionized water. 140 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 750 r / min for 48 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0071] Example 7

[0072] Preparation of cisplatin-polyglutamic acid conjugate

[0073] 200 mg of polyglutamic acid with a relative molecular mass of 2000 Da was dissolved in 36 mL of deionized water. 20 mg of cisplatin was added to the polyglutamic acid solution. The mixture was stirred continuously at 10000 r / min for 24 h at 30 °C in the dark. As the reaction proceeded, the solution color gradually changed from yellow to colorless.

[0074] Example 8

[0075] Preparation of cisplatin-polyglutamic acid nanoparticles

[0076] At room temperature, 1.5 mL of 0.03645 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 1, which was continuously stirred at 1500 r / min. The mixture was stirred at room temperature for 10 min at 1500 r / min. The solid precipitate nanoparticles were collected by centrifugation. The obtained nanoparticles were washed with water by centrifugation and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0077] Transmission electron microscopy image of the cisplatin-polyglutamic acid nanoparticles prepared in Example 8 is shown below. Figure 2 As shown in the figure, the cisplatin-polyglutamic acid nanoparticles have a solid spherical structure with an average particle size of approximately 150 nm.

[0078] Example 9

[0079] Preparation of cisplatin-polyglutamic acid nanoparticles

[0080] At room temperature, 5.5 mL of 0.01 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 1, which was continuously stirred at 1500 r / min. The mixture was stirred at 1500 r / min for 10 min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0081] Example 10

[0082] Preparation of cisplatin-polyglutamic acid nanoparticles

[0083] At room temperature, 1.5 mL of CTAB solution with a concentration of 0.03645 g / mL was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 1, which was continuously stirred at a speed of 1500 r / min. The mixture was stirred at a speed of 1500 r / min for 10 min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0084] Example 11

[0085] Preparation of cisplatin-polyglutamic acid nanoparticles

[0086] At room temperature, 0.55 mL of 0.1 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 1, which was continuously stirred at 1500 r / min. The mixture was stirred at 1500 r / min for 10 min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0087] Example 12

[0088] Preparation of cisplatin-polyglutamic acid nanoparticles

[0089] At room temperature, 1.5 mL of CTAB solution with a concentration of 0.03645 g / mL was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 3, which was continuously stirred at a speed of 750 r / min. The mixture was stirred at a speed of 750 r / min for 20 min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0090] Figure 3 The UV spectrum of the cisplatin-polyglutamic acid nanoparticles prepared in Example 12 is shown below.

[0091] Example 13

[0092] Preparation of cisplatin-polyglutamic acid nanoparticles

[0093] At room temperature, 5.47 mL of 0.01 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 5, which was continuously stirred at 750 r / min. The mixture was stirred at 750 r / min for 20 min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0094] Example 14

[0095] Preparation of cisplatin-polyglutamic acid nanoparticles

[0096] At room temperature, 0.55 mL of 0.1 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 7, which was continuously stirred at 750 r / min. The mixture was stirred at 750 r / min for 20 min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0097] Figure 4 The Fourier transform infrared spectrum of the cisplatin-polyglutamic acid nanoparticles prepared in Example 14 is shown.

[0098] Example 15

[0099] Preparation of cisplatin-polyglutamic acid nanoparticles

[0100] At room temperature, 1.5 mL of CTAB solution with a concentration of 0.03645 g / mL was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 4, which was continuously stirred at a speed of 10 r / min. The mixture was stirred at a speed of 10 r / min for 60 min. The resulting nanoparticles were centrifuged, washed with water, and vacuum dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0101] Example 16

[0102] Preparation of cisplatin-polyglutamic acid nanoparticles

[0103] At room temperature, 5.47 mL of 0.01 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 6, which was continuously stirred at 10 r / min. The mixture was then continuously stirred at 10 r / min. The resulting nanoparticles were centrifuged, washed with water, and freeze-dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0104] Example 17

[0105] Preparation of cisplatin-polyglutamic acid nanoparticles

[0106] At room temperature, 0.55 mL of 0.1 g / mL CTAB solution was rapidly added to the cisplatin-polyglutamic acid conjugate dispersion prepared in Example 7, which was continuously stirred at 10 r / min. The mixture was stirred at 10 r / min for 60 min. The resulting nanoparticles were centrifuged, washed with water, and vacuum dried to obtain cisplatin-polyglutamic acid nanoparticles.

[0107] Example 18

[0108] Preparation of MnO2-cisplatin-polyglutamic acid nanoparticles

[0109] All the cisplatin-polyglutamic acid nanoparticles prepared in Example 8 were centrifuged and redispersed in 36 mL of deionized water. 10 mg of KMnO4 was added to the solution, and the mixture was stirred at 1500 r / min for 10 min at room temperature. The reaction system gradually turned brownish-yellow. The obtained product was centrifuged, washed with water, and freeze-dried to obtain MnO2-cisplatin-polyglutamic acid nanoparticles.

[0110] Figure 5 The UV spectrum of the MnO2-cisplatin-polyglutamic acid nanoparticles prepared in Example 18 is shown below; the transmission electron microscope image of the MnO2-cisplatin-polyglutamic acid nanoparticles prepared in Example 18 is shown below. Figure 6 As shown.

[0111] Example 19

[0112] Preparation of MnO2-cisplatin-polyglutamic acid nanoparticles

[0113] All the cisplatin-polyglutamic acid nanoparticles prepared in Example 12 were centrifuged and redispersed in 36 mL of deionized water. 10 mg of KMnO4 was added to the solution, and the mixture was stirred at 1500 r / min for 10 min at room temperature. The reaction system gradually turned brownish-yellow. The obtained product was centrifuged, washed with water, and freeze-dried to obtain MnO2-cisplatin-polyglutamic acid nanoparticles.

[0114] Example 20

[0115] Preparation of MnO2-cisplatin-polyglutamic acid nanoparticles

[0116] All the cisplatin-polyglutamic acid nanoparticles prepared in Example 16 were centrifuged and redispersed in 36 mL of deionized water. 20 mg of KMnO4 was added to the solution, and the mixture was stirred at 1500 r / min for 10 min at room temperature. The reaction system gradually turned brownish-yellow. The obtained product was centrifuged, washed with water, and vacuum heat-dried to obtain MnO2-cisplatin-polyglutamic acid nanoparticles.

[0117] Example 21

[0118] Preparation of MnO2-cisplatin-polyglutamic acid nanoparticles

[0119] All the cisplatin-polyglutamic acid nanoparticles prepared in Example 14 were centrifuged and redispersed in 36 mL of deionized water. 10 mg of KMnO4 was added to the solution, and the mixture was stirred at 1500 r / min for 10 min at room temperature. The reaction system gradually turned brownish-yellow. The obtained product was centrifuged, washed with water, and vacuum heat-dried to obtain MnO2-cisplatin-polyglutamic acid nanoparticles.

[0120] Example 22

[0121] Human alveolar basal epithelial cells A549 in the logarithmic growth phase were collected and seeded in 6-well plates (2 × 10⁻⁶ cells / well). 5 Cells were cultured on wells (cells / well). After cell adhesion, the culture medium was replaced with fresh medium (2 ml). Cisplatin and the MnO2-cisplatin-polyglutamic acid nanoparticles prepared in Example 18 (containing an equal amount of cisplatin) were added separately, calculated based on the final cisplatin concentration, to achieve molar concentrations of 12.5 μM, 25 μM, 50 μM, and 100 μM for cisplatin and the MnO2-cisplatin-polyglutamic acid nanoparticles (containing an equal amount of cisplatin), respectively. After culturing in a 5% CO2 cell culture incubator for 24 hours (37°C), cells were collected and mixed with V-FITC and PI from the apoptosis detection kit for 30 minutes. The apoptosis rate was detected by flow cytometry. Figure 7 As shown, the MnO2-cisplatin-polyglutamic acid nanoparticles prepared in this invention improve the killing effect on tumor cells by 2-3 times.

Claims

1. A method for preparing a nanoformulation of cisplatin-based anticancer drugs, characterized by, The method comprises the following steps: The polyglutamic acid with a relative molecular mass of 1000 Da is dissolved in 36 mL of deionized water, 140 mg of cisplatin is added into the polyglutamic acid solution, and the solution is continuously stirred at a speed of 1500 r / min for 24 h in dark; 1.5 mL of CTAB solution with a concentration of 0.03645 g / mL is quickly added into the obtained cisplatin-polyglutamic acid combination solution, and the solution is continuously stirred at a speed of 1500 r / min, and the obtained nanoparticles are centrifuged, washed with water, and dried to obtain cisplatin-polyglutamic acid nanoparticles; The nano-preparation of the cisplatin anticancer drug is in a solid spherical structure, and the average particle size is about 150 nm.

2. A method for preparing a nano-formulation for enhancing the efficacy of cisplatin-based anticancer drugs, characterized by, The method comprises the following steps: The cisplatin-polyglutamic acid nanoparticles prepared in claim 1 are centrifuged and dispersed into 36 mL of deionized water, 10 mg of KMnO4 is added into the solution, the solution is stirred at a speed of 1500 r / min at room temperature for 10 min, the reaction system is gradually changed into brown yellow, and the obtained product is centrifuged, washed with water, and freeze-dried to obtain MnO2-cisplatin-polyglutamic acid nano-preparation, which is the nano-preparation for enhancing the therapeutic effect of the cisplatin anticancer drug; the thickness of the MnO2 shell layer is 0.01 nm-50 nm.

3. The nano-preparation of the cisplatin anticancer drug prepared by the preparation method of the nano-preparation of the cisplatin anticancer drug in claim 1.

4. The nano-preparation for enhancing the therapeutic effect of the cisplatin anticancer drug prepared by the preparation method of the nano-preparation for enhancing the therapeutic effect of the cisplatin anticancer drug in claim 2.

5. The nano-preparation of the cisplatin anticancer drug in claim 3 or the nano-preparation for enhancing the therapeutic effect of the cisplatin anticancer drug in claim 4 in the preparation of a drug for treating tumors.

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