Composition for treating gastric cancer as well as preparation method and application thereof

Abstract

The invention provides a composition for treating gastric cancer as well as a preparation method and application thereof, and belongs to the technical field of medicines. The preparation method comprises the following steps: preparing composite nanoparticles with boron nitride deposited on gold nanocages, modifying the surfaces of the composite nanoparticles with dendrobium officinale polysaccharides, loading garlic saponin / dioscin / paclitaxel eutectic crystals to obtain a compound, and carrying out self-assembly reaction on extracted HGC-27 human gastric cancer cell membrane protein and the compound to obtain the composition for treating gastric cancer. The composition for treating gastric cancer, prepared by the invention, has the characteristics of good biocompatibility and biological safety, targeted drug delivery, small side effect, obviously improved drug effect, slow release characteristic, improved patient compliance, good treatment effect on gastric cancer, and wide application prospect.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a composition for treating gastric cancer, and a preparation method and application thereof. Background Art

[0002] Gastric cancer (GC) is one of the most common cancers and seriously endangers the health of residents. Reducing the morbidity and mortality of gastric cancer is increasingly becoming a major public health issue that we must face.

[0003] The occurrence of gastric cancer is the result of the combined effects of internal and external factors such as genetics and the environment. In terms of genetic factors, family history is an important risk factor for the onset of gastric cancer. In recent years, the application of molecular biology technology has established the relationship between gastric cancer and family history. In terms of environmental factors, bad dietary habits, smoking, and Helicobacter pylori are all risk factors for inducing gastric cancer. Due to the low rate of early diagnosis, most patients are already in the middle and late stages of the disease when diagnosed. Traditional cytotoxic chemotherapy drugs are reliable for malignant tumors, but due to serious adverse reactions such as gastrointestinal reactions and bone marrow suppression, their clinical application is limited and the quality of life of patients is affected. Molecular targeted therapy is a new treatment method that has emerged in the treatment of hematological tumors and solid tumors in recent years. With the gradual deepening of research on the molecular biological mechanisms of gastric cancer occurrence, development and metastasis, this treatment method has also been gradually applied to the clinical practice of gastric cancer treatment.

[0004] Cancer treatment is extremely difficult, and it is still a technical problem that the medical community is working hard to solve. Modern medicine uses surgery, chemotherapy, and radiotherapy as treatment methods. Among them, surgical treatment is prone to cause complications such as bleeding and perforation, and radiotherapy is prone to cause complications such as radiation myelopathy. In short, the treatment methods of surgery and radiotherapy have large side effects, a very high probability of recurrence and metastasis, and a low survival rate. In addition, there are many drugs for treating cancer, but the effect is poor, mainly manifested in low efficacy or large toxic side effects, and it is difficult to extend the patient's survival period. The reason is that many drugs do not fundamentally solve the problem of cancer. Summary of the invention

[0005] The purpose of the present invention is to provide a composition for treating gastric cancer, a preparation method and an application thereof, which has good biocompatibility and biosafety, has the characteristics of targeted drug delivery, has small side effects, significantly improved drug efficacy, has sustained-release properties, improves patient compliance, has a good therapeutic effect on gastric cancer, and has broad application prospects.

[0006] The technical solution of the present invention is achieved in this way: The invention provides a method for preparing a composition for treating gastric cancer. The method comprises the following steps: preparing composite nanoparticles with boron nitride deposited on gold nanocages, modifying the surface of the composite nanoparticles with dendrobium officinale polysaccharide, loading garlic saponin / dioscin / paclitaxel cocrystals to obtain a composite, and then subjecting the extracted HGC-27 human gastric cancer cell membrane protein to a self-assembly reaction with the composite to obtain a composition for treating gastric cancer.

[0007] As a further improvement of the present invention, the following steps are included: S1. Preparation of boron nitride / gold nanoparticles: adding gold nanocages to a solvent, adding sodium borohydride and ammonium formate, heating and stirring to react, removing the solvent under reduced pressure, washing the solid, drying, and calcining to obtain boron nitride / gold nanoparticles; S2. Preparation of drug co-crystals: Wash garlic and yam, dry, crush, add to a deep eutectic solvent, heat and stir to extract, add dichloromethane to precipitate, wash the solid, dry, and dissolve in ethanol to obtain solution A; dissolve paclitaxel in a mixed solvent to obtain solution B, mix solution A and solution B evenly, evaporate the solvent, precipitate crystals, filter, wash, and dry to obtain drug co-crystals; S3. Preparation of Dendrobium officinale polysaccharide: Wash the Dendrobium officinale, dry it, crush it, add it to water, heat it to boiling for extraction, filter it, add ethanol to the filtrate for precipitation, wash the precipitate, and dry it to obtain Dendrobium officinale polysaccharide; S4. Hydrophilic modification: mixing boron nitride / gold nanoparticles and Dendrobium officinale polysaccharide and ball milling to prepare modified boron nitride / gold nanoparticles; S5. Drug loading: ball-mill the drug co-crystal, add it to water, add the modified boron nitride / gold nanoparticles, stir and mix evenly, centrifuge, wash, and dry to obtain drug-loaded nanoparticles; S6. Targeted modification: HGC-27 human gastric cancer cells in the logarithmic growth phase were digested and collected, membrane proteins were extracted using a membrane protein extraction kit, a membrane protein solution was prepared, drug-loaded nanoparticles were added, self-assembled by ultrasonic treatment, the reaction was stirred, and freeze-dried to obtain a composition for treating gastric cancer.

[0008] As a further improvement of the present invention, the mass ratio of the gold nanocage, sodium borohydride and ammonium formate in step S1 is 10:3-5:10-12, the temperature of the heating and stirring reaction is 45-55°C, the time is 1-2h, and the calcination temperature is 1000-1200°C, and the time is 3-5h.

[0009] As a further improvement of the present invention, the mass ratio of garlic, yam and deep eutectic solvent in step S2 is 5-7:3-5:50-70, the temperature of the heating and stirring extraction is 60-70°C, and the time is 0.5-1.5h; the mass ratio of the solid and paclitaxel is 3-5:0.5-1, and the mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol in a volume ratio of 3-5:3-5; the deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 1-3:1, heating to 70-80°C, and cooling to room temperature.

[0010] As a further improvement of the present invention, the solid-liquid ratio of Dendrobium officinale and water in step S3 is 1:5-10 g / mL, and the extraction time is 2-4 h.

[0011] As a further improvement of the present invention, in step S4, the mass ratio of boron nitride / gold nanoparticles to Dendrobium officinale polysaccharide is 7-10:3-4, and the ball milling time is 1-2h.

[0012] As a further improvement of the present invention, the ball milling time in step S5 is 0.5-1h, and the mass ratio of the drug cocrystal to modified boron nitride / gold nanoparticles is 3-5:10.

[0013] As a further improvement of the present invention, the mass ratio of the membrane protein solution and the drug-loaded nanoparticles in step S6 is 100:15-22, the concentration of the membrane protein solution is 5-10 mg / mL, the power of the ultrasonic self-assembly is 500-1000 W, the time is 10-20 min, and the stirring reaction is 8-10 h.

[0014] The present invention further protects a composition for treating gastric cancer prepared by the above preparation method.

[0015] The present invention further protects the use of the above-mentioned composition for treating gastric cancer in preparing a drug for treating and / or preventing gastric cancer.

[0016] The present invention has the following beneficial effects: The invention uses gold nano cages as carriers, loads sodium borohydride and ammonium formate, reacts to generate ammonia borane products, further calcines and cracks, and obtains boron nitride / gold nanoparticles. The gold nano cages have good biocompatibility, improved cell adhesion, and good biosafety. At the same time, the presence of the gold component enables the prepared composition to fix membrane proteins on the surface of the composite material through thiol groups. At the same time, the gold component can also form a gold-DNA complex with the DNA sequence of the cancer cell by means of the thiol group, so that the drug molecules are inserted into the cancer cell DNA. Under the good near-infrared light response characteristics of the nano gold, under the stimulation of the near-infrared light, the temperature of the gold surface will increase, the hydrogen bonds of the DNA chain will open and release the drugs into the cells, and the drug concentration in the tumor site will be increased to a certain extent, avoiding the drug resistance problem of direct use of chemotherapy, and improving the chemotherapy effect.

[0017] However, the hydrophilicity of boron nitride materials is poor, which limits their application. The present invention blends the prepared Dendrobium officinale polysaccharide with boron nitride / gold nanoparticles for ball milling, so that the boron nitride / gold nanoparticles are modified by van der Waals forces such as hydrogen bonds with the polysaccharide, greatly enhancing the hydrophilicity of the boron nitride / gold nanoparticles, making it easier to release drugs, and at the same time, under the modification of the boron nitride / gold nanoparticles by the Dendrobium officinale polysaccharide, the Dendrobium officinale polysaccharide can polarize THP-1 macrophages from M2 type to M1 type, downregulate the expression of signal transduction and transcription activator factor 6 / peroxisome proliferator-activated body receptor-γ pathway and type 1 arginase and transglutaminase 2 protein, inhibit the JAGGED1 / NOTCH1 signaling pathway, promote gastric cancer cell apoptosis and inhibit migration, thereby further improving the anti-tumor performance of the material.

[0018] The present invention heats garlic and yam through deep eutectic solvent extraction, with high extraction rate, and obtains garlic saponin and dioscin through organic solvent precipitation. Dioscin is a natural flavonoid active ingredient, which inhibits M2 macrophage polarization through TRAF2 / NF-κB pathway, and plays a role in inhibiting gastric cancer invasion and growth; garlic saponin downregulates TGF-β protein expression through targeted downregulation, upregulates Smad4 protein expression, improves the mutation state of Smad4, plays its role in regulating transport, combines with activated receptor-regulated Smads protein, produces a oligomeric complex, plays a major role in the TGF-B / Smads signal transduction process, can inhibit proto-oncogene expression, reduce the formation of tumor cells, thereby improving gastric mucosal pathological changes, and preventing the occurrence and development of gastric precancerous lesions. The two have a synergistic effect. Paclitaxel is a highly active anticancer chemotherapy drug, but it has certain blood and neurotoxicity, and its water solubility is extremely poor, which greatly limits its application. If paclitaxel is used in large quantities, it will cause great harm to the human body. However, in the traditional treatment scheme, the effect of a small amount of paclitaxel is poor and cannot play a good anti-cancer effect. The present invention prepares a garlic saponin / dioscin / paclitaxel cocrystal, and the natural saponin and paclitaxel are compounded to form a cocrystal, which greatly improves the water solubility of the product on the one hand, and on the other hand, under the synergistic anti-cancer effect of garlic saponin and dioscin, the amount of paclitaxel used is greatly reduced, thereby effectively reducing the occurrence of side effects and improving patient compliance. Moreover, the formed cocrystal structure also reduces the occurrence of some side effects to a certain extent.

[0019] The role of the tumor microenvironment in tumorigenesis is increasingly being explored. During the growth of gastric cancer, mesenchymal stem cells are recruited into the tumor microenvironment of gastric cancer, and they change the tumor microenvironment and promote tumor growth by secreting multiple factors. The cell membrane-wrapped nanocomplex not only has good biocompatibility, but also has the advantages of long in vivo circulation time, targeting, immune escape, controlled drug release, immune activation, etc. The present invention separates and obtains gastric cancer membrane proteins, and loads them on the surface of drug-loaded nanoparticles, effectively avoiding their clearance by the immune system, prolonging their drug action time, avoiding drug circulation to other organs, reducing the side effects of drugs, and can achieve targeted delivery of drugs to tumor tissues through orderly targeting mediated by membrane proteins and tumor cell membrane proteins, thereby improving drug efficacy.

[0020] The composition for treating gastric cancer prepared by the present invention has good biocompatibility and biosafety, has the characteristics of targeted drug delivery, has small side effects, significantly improved drug efficacy, has sustained-release properties, improves patient compliance, has a good therapeutic effect on gastric cancer, and has broad application prospects. DETAILED DESCRIPTION

[0021] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0022] Membrane protein extraction kit, brand: Solarbio, product number EX1110.

[0023] HGC-27 human gastric cancer cells were purchased from Wuhan Sun Biotechnology Co., Ltd.

[0024] Gold nanocages, with an average particle size of 100 nm, were purchased from Nanjing Dongna Biotechnology Co., Ltd. Example 1

[0025] This embodiment provides a method for preparing a composition for treating gastric cancer, comprising the following steps: S1. Preparation of boron nitride / gold nanoparticles: 10 g of gold nanocages were added to 250 mL of 1,4-dioxane, 3 g of sodium borohydride and 10 g of ammonium formate, heated to 45°C, stirred for reaction for 1 h, 1,4-dioxane was removed under reduced pressure, the solid was washed, dried, and calcined at 1000°C for 3 h to obtain boron nitride / gold nanoparticles; S2. Preparation of drug cocrystals: 5 g of garlic and 3 g of yam were washed, dried, crushed, added to 50 g of deep eutectic solvent, heated to 60 ° C, stirred and extracted for 0.5 h, added to 100 mL of dichloromethane for precipitation, the precipitate was washed and dried to obtain a solid, 0.6 g of the solid was dissolved in 50 mL of ethanol to obtain solution A; 0.1 g of paclitaxel was dissolved in 20 mL of a mixed solvent to obtain solution B, solution A and solution B were mixed, the solvent was evaporated for 2 days, crystals were precipitated, filtered, washed, and dried to obtain drug cocrystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 3:5; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 1:1, heating to 70° C., and cooling to room temperature; S3. Preparation of Dendrobium officinale polysaccharide: 10 g of Dendrobium officinale was washed, dried, crushed, added to 50 mL of water, heated to boiling and extracted for 2 h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 90 wt %, precipitated for 1 h, the precipitate was washed and dried to obtain Dendrobium officinale polysaccharide; S4. Hydrophilic modification: 7 g of boron nitride / gold nanoparticles and 3 g of Dendrobium officinale polysaccharide were mixed and ball-milled for 1 h to obtain modified boron nitride / gold nanoparticles; S5. Drug loading: 3 g of drug cocrystals were ball-milled for 0.5 h, added to 100 mL of water, 10 g of modified boron nitride / gold nanoparticles were added, stirred and mixed for 1 h, centrifuged, washed, and dried to obtain drug-loaded nanoparticles; S6. Targeted modification: HGC-27 human gastric cancer cells in the logarithmic growth phase were digested and collected, membrane proteins were extracted using a membrane protein extraction kit, and a 5 mg / mL membrane protein solution was prepared. 15 g of drug-loaded nanoparticles were added to 100 mL of the membrane protein solution, and self-assembly was performed at 500 W ultrasonic treatment for 10 min. The reaction was stirred for 8 h, and freeze-dried to obtain a composition for the treatment of gastric cancer. Example 2

[0026] This embodiment provides a method for preparing a composition for treating gastric cancer, comprising the following steps: S1. Preparation of boron nitride / gold nanoparticles: 10 g of gold nanocages were added to 250 mL of 1,4-dioxane, 5 g of sodium borohydride and 12 g of ammonium formate, heated to 55°C, stirred for reaction for 2 h, 1,4-dioxane was removed under reduced pressure, the solid was washed, dried, and calcined at 1200°C for 5 h to obtain boron nitride / gold nanoparticles; S2. Preparation of drug cocrystals: 7g of garlic and 5g of yam were washed, dried, crushed, added to 70g of deep eutectic solvent, heated to 70°C, stirred and extracted for 1.5h, 100mL of dichloromethane was added for precipitation, the precipitate was washed, dried to obtain a solid, 1g of the solid was dissolved in 50mL of ethanol to obtain solution A; 0.2g of paclitaxel was dissolved in 20mL of a mixed solvent to obtain solution B, solution A and solution B were mixed, the solvent was evaporated for 2d, crystals were precipitated, filtered, washed, and dried to obtain drug cocrystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 5:3; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 3:1, heating to 80° C., and cooling to room temperature; S3. Preparation of Dendrobium officinale polysaccharide: 10g of Dendrobium officinale was washed, dried, crushed, added to 100mL of water, heated to boiling and extracted for 4h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 90wt%, precipitated for 1h, the precipitate was washed and dried to obtain Dendrobium officinale polysaccharide; S4. Hydrophilic modification: 10 g of boron nitride / gold nanoparticles and 4 g of Dendrobium officinale polysaccharide were mixed and ball-milled for 2 h to obtain modified boron nitride / gold nanoparticles; S5. Drug loading: 5 g of drug cocrystals were ball-milled for 1 h, added to 100 mL of water, 10 g of modified boron nitride / gold nanoparticles were added, stirred and mixed for 1 h, centrifuged, washed, and dried to obtain drug-loaded nanoparticles; S6. Targeted modification: HGC-27 human gastric cancer cells in the logarithmic growth phase were digested and collected, membrane proteins were extracted using a membrane protein extraction kit, and a 10 mg / mL membrane protein solution was prepared. 22 g of drug-loaded nanoparticles were added to 100 mL of the membrane protein solution, and self-assembly was performed at 1000 W ultrasonic treatment for 20 min. The reaction was stirred for 10 h, and freeze-dried to obtain a composition for the treatment of gastric cancer. Example 3

[0027] This embodiment provides a method for preparing a composition for treating gastric cancer, comprising the following steps: S1. Preparation of boron nitride / gold nanoparticles: 10 g of gold nanocages were added to 250 mL of 1,4-dioxane, 4 g of sodium borohydride and 11 g of ammonium formate, heated to 50°C, stirred for reaction for 1.5 h, 1,4-dioxane was removed under reduced pressure, the solid was washed, dried, and calcined at 1100°C for 4 h to obtain boron nitride / gold nanoparticles; S2. Preparation of drug cocrystals: 6 g of garlic and 4 g of yam were washed, dried, crushed, added to 60 g of deep eutectic solvent, heated to 65 ° C, stirred and extracted for 1 h, added with 100 mL of dichloromethane for precipitation, the precipitate was washed and dried to obtain a solid, 0.8 g of the solid was dissolved in 50 mL of ethanol to obtain solution A; 0.15 g of paclitaxel was dissolved in 20 mL of a mixed solvent to obtain solution B, solution A and solution B were mixed, the solvent was evaporated for 2 days, crystals were precipitated, filtered, washed, and dried to obtain drug cocrystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 1:1; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 2:1, heating to 75° C., and cooling to room temperature; S3. Preparation of Dendrobium officinale polysaccharide: 10 g of Dendrobium officinale was washed, dried, crushed, added to 75 mL of water, heated to boiling and extracted for 3 h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 90 wt %, precipitated for 1 h, the precipitate was washed and dried to obtain Dendrobium officinale polysaccharide; S4. Hydrophilic modification: 8.5 g of boron nitride / gold nanoparticles and 3.5 g of Dendrobium officinale polysaccharide were mixed and ball-milled for 1.5 h to obtain modified boron nitride / gold nanoparticles; S5. Drug loading: 4 g of drug cocrystals were ball-milled for 1 h, added to 100 mL of water, 10 g of modified boron nitride / gold nanoparticles were added, stirred and mixed for 1 h, centrifuged, washed, and dried to obtain drug-loaded nanoparticles; S6. Targeted modification: HGC-27 human gastric cancer cells in the logarithmic growth phase were digested and collected, membrane proteins were extracted using a membrane protein extraction kit, and a 7 mg / mL membrane protein solution was prepared. 19 g of drug-loaded nanoparticles were added to 100 mL of the membrane protein solution, and self-assembled by 700 W ultrasonic treatment for 15 min. The reaction was stirred for 9 h, and freeze-dried to obtain a composition for the treatment of gastric cancer.

[0028] Comparative Example 1 Compared with Example 3, the difference is that no gold nanocages are added in step S1.

[0029] The details are as follows: S1. Preparation of boron nitride nanoparticles: 4 g of sodium borohydride and 11 g of ammonium formate were added to 250 mL of 1,4-dioxane, heated to 50°C, stirred for reaction for 1.5 h, 1,4-dioxane was removed under reduced pressure, the solid was washed, dried, and calcined at 1100°C for 4 h to obtain boron nitride nanoparticles.

[0030] Comparative Example 2 Compared with Example 3, the difference is that garlic is not added in step S2.

[0031] The details are as follows: S2. Preparation of drug co-crystals: 10 g of Dioscorea was washed, dried, crushed, added to 60 g of deep eutectic solvent, heated to 65 ° C, stirred and extracted for 1 h, 100 mL of dichloromethane was added for precipitation, the precipitate was washed, dried to obtain a solid, 0.8 g of the solid was dissolved in 50 mL of ethanol to obtain solution A; 0.15 g of paclitaxel was dissolved in 20 mL of a mixed solvent to obtain solution B, solution A and solution B were mixed, the solvent was evaporated for 2 days, crystals were precipitated, filtered, washed, and dried to obtain drug co-crystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 1:1; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 2:1, heating to 75° C., and cooling to room temperature.

[0032] Comparative Example 3 Compared with Example 3, the difference is that Dioscorea zingiberensis is not added in step S2.

[0033] The details are as follows: S2. Preparation of drug cocrystals: Wash, dry, and crush 10 g of garlic, add it to 60 g of deep eutectic solvent, heat it to 65 ° C, stir and extract for 1 h, add 100 mL of dichloromethane to precipitate, wash the precipitate, and dry it to obtain a solid. Dissolve 0.8 g of the solid in 50 mL of ethanol to obtain solution A; dissolve 0.15 g of paclitaxel in 20 mL of a mixed solvent to obtain solution B, mix solution A and solution B, evaporate the solvent for 2 days, precipitate crystals, filter, wash, and dry to obtain drug cocrystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 1:1; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 2:1, heating to 75° C., and cooling to room temperature.

[0034] Comparative Example 4 Compared with Example 3, the difference is that in step S2, the solid and paclitaxel are simply mixed.

[0035] The details are as follows: S2. Preparation of drug cocrystals: 6 g of garlic and 4 g of yam were washed, dried, crushed, added to 60 g of deep eutectic solvent, heated to 65 ° C, stirred and extracted for 1 h, added with 100 mL of dichloromethane for precipitation, the precipitate was washed and dried to obtain a solid, 0.8 g of the solid and 0.15 g of paclitaxel were mixed evenly to obtain a mixture; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 1:1; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 2:1, heating to 75° C., and cooling to room temperature.

[0036] Comparative Example 5 Compared with Example 3, the difference is that steps S3 and S4 are not performed.

[0037] The details are as follows: S1. Preparation of boron nitride / gold nanoparticles: 10 g of gold nanocages were added to 250 mL of 1,4-dioxane, 4 g of sodium borohydride and 11 g of ammonium formate, heated to 50°C, stirred for reaction for 1.5 h, 1,4-dioxane was removed under reduced pressure, the solid was washed, dried, and calcined at 1100°C for 4 h to obtain boron nitride / gold nanoparticles; S2. Preparation of drug cocrystals: 6 g of garlic and 4 g of yam were washed, dried, crushed, added to 60 g of deep eutectic solvent, heated to 65 ° C, stirred and extracted for 1 h, added with 100 mL of dichloromethane for precipitation, the precipitate was washed and dried to obtain a solid, 0.8 g of the solid was dissolved in 50 mL of ethanol to obtain solution A; 0.15 g of paclitaxel was dissolved in 20 mL of a mixed solvent to obtain solution B, solution A and solution B were mixed, the solvent was evaporated for 2 days, crystals were precipitated, filtered, washed, and dried to obtain drug cocrystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 1:1; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 2:1, heating to 75° C., and cooling to room temperature; S3. Drug loading: 4 g of drug cocrystals were ball-milled for 1 h, added to 100 mL of water, 10 g of boron nitride / gold nanoparticles were added, stirred and mixed for 1 h, centrifuged, washed, and dried to obtain drug-loaded nanoparticles; S4. Targeted modification: HGC-27 human gastric cancer cells in the logarithmic growth phase were digested and collected, membrane proteins were extracted using a membrane protein extraction kit, and a 7 mg / mL membrane protein solution was prepared. 19 g of drug-loaded nanoparticles were added to 100 mL of the membrane protein solution, and self-assembly was performed at 700 W ultrasonic treatment for 15 min. The reaction was stirred for 9 h, and freeze-dried to obtain a composition for the treatment of gastric cancer.

[0038] Comparative Example 6 Compared with Example 3, the difference is that in step S5, the drug cocrystal is replaced by an equal amount of paclitaxel.

[0039] The details are as follows: S5. Drug loading: 4 g of paclitaxel was added to 100 mL of water, and 10 g of modified boron nitride / gold nanoparticles were added. The mixture was stirred for 1 h, centrifuged, washed, and dried to obtain drug-loaded nanoparticles.

[0040] Comparative Example 7 Compared with the embodiment 3, the difference is that step S6 is not performed.

[0041] The details are as follows: S1. Preparation of boron nitride / gold nanoparticles: 10 g of gold nanocages were added to 250 mL of 1,4-dioxane, 4 g of sodium borohydride and 11 g of ammonium formate, heated to 50°C, stirred for reaction for 1.5 h, 1,4-dioxane was removed under reduced pressure, the solid was washed, dried, and calcined at 1100°C for 4 h to obtain boron nitride / gold nanoparticles; S2. Preparation of drug cocrystals: 6 g of garlic and 4 g of yam were washed, dried, crushed, added to 60 g of deep eutectic solvent, heated to 65 ° C, stirred and extracted for 1 h, added with 100 mL of dichloromethane for precipitation, the precipitate was washed and dried to obtain a solid, 0.8 g of the solid was dissolved in 50 mL of ethanol to obtain solution A; 0.15 g of paclitaxel was dissolved in 20 mL of a mixed solvent to obtain solution B, solution A and solution B were mixed, the solvent was evaporated for 2 days, crystals were precipitated, filtered, washed, and dried to obtain drug cocrystals; The mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 1:1; The deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 2:1, heating to 75° C., and cooling to room temperature; S3. Preparation of Dendrobium officinale polysaccharide: 10 g of Dendrobium officinale was washed, dried, crushed, added to 75 mL of water, heated to boiling and extracted for 3 h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 90 wt %, precipitated for 1 h, the precipitate was washed and dried to obtain Dendrobium officinale polysaccharide; S4. Hydrophilic modification: 8.5 g of boron nitride / gold nanoparticles and 3.5 g of Dendrobium officinale polysaccharide were mixed and ball-milled for 1.5 h to obtain modified boron nitride / gold nanoparticles; S5. Drug loading: 4 g of drug cocrystals were ball-milled for 1 h, added into 100 mL of water, 10 g of modified boron nitride / gold nanoparticles were added, stirred and mixed for 1 h, centrifuged, washed, and dried to obtain drug-loaded nanoparticles, which are the composition for treating gastric cancer.

[0042] Test Example 1 The composition for treating gastric cancer prepared in Example 1-3 or Comparative Example 1-7 was prepared into a 150 μg / mL suspension using a PBS solution with a pH of 7 as a sample solution.

[0043] THLE-3 cells (liver epithelial cells) and HGC-27 cells (human gastric cancer cells) were placed in DMEM culture medium and cultured at 37°C, CO2 5v / v% for 12 h. Subsequently, THLE-3 cells and HGC-27 cells were seeded in 96-well plates, and the cell density was controlled to be (1-2)×10 5 Each well was cultured for 24 hours, and then 200 μL of sample solution was added to treat the cells for 24 hours, and near-infrared light (808 nm) was used to irradiate the tumor for 5 minutes. Deionized water was used as a blank control, and finally, the relative survival rate of the cells relative to the control group was determined according to the operating procedures of the MTT kit. The results are shown in Table 1.

[0044] Table 1

[0045] It can be seen from the above table that the compositions for treating gastric cancer prepared in Examples 1-3 of the present invention have lower cytotoxicity, but a high killing rate for gastric cancer cells.

[0046] Test Example 2 Wistar rats were selected, SPF grade, half male and half female, with a body weight of 55-70 g. Human gastric cancer cell SNU-1 cells (ATCC® CRL-5971, USA) in the logarithmic growth phase were prepared at a concentration of 5×10 6 / ml of cell solution. Subsequently, the rat's forelimb (left side) was subcutaneously injected (injection volume of 0.8mL). In this way, cells can proliferate and form tumors in subcutaneous tissue. On the 7th day after the modeling process, if the presence of the tumor can be detected by touch, it can be confirmed that the tumor model has been successfully established. The rats with successful modeling were randomly divided into a model group, a positive drug group, an embodiment 1-3 group and a comparative example 1-7 group, with 10 rats in each group. The embodiment 1-3 group and the comparative example 1-7 group were given 20mg / kg of the corresponding composition for treating gastric cancer, the positive drug group was given 1mg / kg of cisplatin, and the blank group and the model group were given an equal amount of normal saline. After the administration was completed, the tumor was irradiated with near-infrared light (808nm) for 5min. After 28d of intervention, the width and length of the tumor were accurately measured using a vernier caliper, and the volume of the tumor was calculated according to the formula volume = 0.5236×〔(width×length) / 2〕, and the tumor inhibition rate was calculated. Tumor growth inhibition rate (%) = [1-T (tumor volume of treatment group) / C (tumor volume of model group)] × 100%. The results are shown in Table 2.

[0047] Table 2

[0048] It can be seen from the above table that the compositions for treating gastric cancer prepared in Examples 1-3 of the present invention have good tumor inhibition effects.

[0049] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a composition for treating gastric cancer, characterized in that: Composite nanoparticles with boron nitride deposited on gold nanocages were prepared, the surface of which was modified with Dendrobium officinale polysaccharide and loaded with garlic saponin / dioscin / paclitaxel cocrystals to obtain a complex, and then the extracted HGC-27 human gastric cancer cell membrane protein was self-assembled with the complex to obtain a composition for treating gastric cancer.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: S1. Preparation of boron nitride / gold nanoparticles: adding gold nanocages to a solvent, adding sodium borohydride and ammonium formate, heating and stirring to react, removing the solvent under reduced pressure, washing the solid, drying, and calcining to obtain boron nitride / gold nanoparticles; S2. Preparation of drug co-crystals: Wash garlic and yam, dry, crush, add to a deep eutectic solvent, heat and stir to extract, add dichloromethane to precipitate, wash the precipitate, dry, dissolve the solid in ethanol to obtain solution A; dissolve paclitaxel in a mixed solvent to obtain solution B, mix solution A and solution B evenly, evaporate the solvent, precipitate crystals, filter, wash, and dry to obtain drug co-crystals; S3. Preparation of Dendrobium officinale polysaccharide: Wash the Dendrobium officinale, dry it, crush it, add it to water, heat it to boiling for extraction, filter it, add ethanol to the filtrate for precipitation, wash the precipitate, and dry it to obtain Dendrobium officinale polysaccharide; S4. Hydrophilic modification: mixing boron nitride / gold nanoparticles and Dendrobium officinale polysaccharide and ball milling to prepare modified boron nitride / gold nanoparticles; S5. Drug loading: ball-mill the drug co-crystal, add it to water, add the modified boron nitride / gold nanoparticles, stir and mix evenly, centrifuge, wash, and dry to obtain drug-loaded nanoparticles; S6. Targeted modification: HGC-27 human gastric cancer cells in the logarithmic growth phase were digested and collected, membrane proteins were extracted using a membrane protein extraction kit, a membrane protein solution was prepared, drug-loaded nanoparticles were added, self-assembled by ultrasonic treatment, the reaction was stirred, and freeze-dried to obtain a composition for treating gastric cancer.

3. The preparation method according to claim 2, characterized in that: In step S1, the mass ratio of the gold nanocage, sodium borohydride and ammonium formate is 10:3-5:10-12, the temperature of the heating and stirring reaction is 45-55°C, the time is 1-2h, and the temperature of the calcination is 1000-1200°C, the time is 3-5h.

4. The preparation method according to claim 2, characterized in that: In step S2, the mass ratio of garlic, yam and deep eutectic solvent is 5-7:3-5:50-70, the temperature of the heating and stirring extraction is 60-70°C, and the time is 0.5-1.5h; the mass ratio of the solid and paclitaxel is 3-5:0.5-1, and the mixed solvent is a mixed solution of polyoxyethylene castor oil and anhydrous ethanol, with a volume ratio of 3-5:3-5; the deep eutectic solvent is a liquid formed by mixing glucose and choline chloride in a molar ratio of 1-3:1, heating to 70-80°C, and cooling to room temperature.

5. The preparation method according to claim 2, characterized in that: In step S3, the solid-to-liquid ratio of Dendrobium officinale and water is 1:5-10 g / mL, and the extraction time is 2-4 h.

6. The preparation method according to claim 2, characterized in that: In step S4, the mass ratio of boron nitride / gold nanoparticles to Dendrobium officinale polysaccharide is 7-10:3-4, and the ball milling time is 1-2h.

7. The preparation method according to claim 2, characterized in that: The ball milling time in step S5 is 0.5-1h, and the mass ratio of the drug cocrystal to modified boron nitride / gold nanoparticles is 3-5:

10.

8. The preparation method according to claim 2, characterized in that: In step S6, the mass ratio of the membrane protein solution to the drug-loaded nanoparticles is 100:15-22, the concentration of the membrane protein solution is 5-10 mg / mL, the power of the ultrasonic self-assembly is 500-1000 W, the time is 10-20 min, and the stirring reaction time is 8-10 h.

9. A composition for treating gastric cancer prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the composition for treating gastric cancer according to claim 9 in preparing a drug for treating and / or preventing gastric cancer.

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