Oxaliplatin multifunctional liposome and preparation method and application thereof

Abstract

The application discloses a kind of oxaliplatin multifunctional liposome and preparation method and application, the liposome is made of oxaliplatin, calcium peroxide nanoparticle and lecithin and ginsenoside Rg3.The oxaliplatin of the present application is co-loaded with calcium peroxide nanoparticle multifunctional liposome, wherein peroxide nanoparticle improves solid tumor hypoxic microenvironment, improves the antitumor effect of oxaliplatin;Ginsenoside Rg3 replaces cholesterol and plays a targeting role, simultaneously as a good chemotherapy adjuvant, combined with chemotherapy drugs to increase the antitumor effect and reduce systemic toxicity;The IC 50 Value of HCT116 cell of the liposome is 5.37±0.42 μM, and the tumor inhibition rate is 87.55±9.35%, with good anti-colorectal cancer effect.

Description

Technical Field

[0001] The present invention relates to a co-loaded multifunctional liposome, a preparation method and an application thereof, and in particular to a co-loaded multifunctional liposome of oxaliplatin and calcium peroxide nanoparticles, a preparation method and an application thereof. Background Art

[0002] Colorectal cancer (CRC) is a heterogeneous disease across tumor anatomical locations and is a multifactorial disease caused by lifestyle, genetics, and environmental factors. Among them, genetic and environmental factors play an important role in the etiology of colorectal cancer. Generally, colorectal cancer subtypes are defined based on the anatomical location of the tumor in the three segments of the colorectum, and are divided into proximal colon cancer (cecum, ascending colon, hepatic flexure, and transverse colon), distal colon cancer (splenic flexure, descending colon, and sigmoid colon), and rectal cancer.

[0003] Platinum-based drugs are the mainstay of clinical anti-tumor therapy. Oxaliplatin (OXA), a third-generation platinum-based anti-cancer drug developed by Debiopharm in Switzerland, exhibits broad-spectrum anti-tumor activity both in vitro and in vivo, but its therapeutic efficacy is inferior to that of cisplatin and carboplatin, and it is associated with adverse reactions such as neurotoxicity and myelosuppression. Furthermore, oxaliplatin is a first-line chemotherapy drug for colorectal cancer, killing tumor cells by interfering with DNA replication and transcription, leading to DNA damage. However, the clinical application of OXA is limited by the unique microenvironment of solid tumors, such as hypoxia, acidosis, and elevated glutathione levels. Recently, various metal peroxides, including calcium peroxide (CaO2), manganese dioxide (MnO2), magnesium peroxide (MgO2), and zinc peroxide (ZnO2), have been investigated for their potential to improve the hypoxic tumor microenvironment by reacting with H2O in tumor tissue to generate oxygen, thereby enhancing the efficacy of chemotherapy. Among these metal peroxides, CaO2, with its high biocompatibility and efficient oxygen-generating capacity, is the most promising material for modulating tumor hypoxia.

[0004] However, because CaO2 is unstable in water and OXA is systemically toxic when administered directly, a drug carrier system needs to be designed for the delivery of CaO2 and OXA. Liposomes are drug carriers with high biocompatibility, low toxicity, and simple preparation. Cholesterol is a basic component of liposomes and has the ability to stabilize the phospholipid bilayer, but cholesterol may cause hyperlipidemia and allergic reactions. Therefore, it is important to find structurally similar cholesterol substitutes to stabilize lipid membranes. Ginsenoside Rg3 is a steroid compound isolated from ginseng that has a lipophilic steroid structure similar to cholesterol and can be used to replace cholesterol. In addition, Rg3 has the ability to actively target glucose transporter 1 (Glut1) overexpressed on the surface of colorectal cancer cells and assist in chemotherapy, and can replace cholesterol as a membrane stabilizer to prepare multifunctional liposomes. Summary of the Invention

[0005] The purpose of the present invention is to provide a multifunctional liposome co-loaded with oxaliplatin and calcium peroxide nanoparticles, as well as a preparation method and application, which can target glucose transporter 1 on the surface of HCT116 cells, increase the targeting and uptake of liposomes, and at the same time regulate the tumor hypoxic microenvironment, enhance the killing effect of chemotherapy drugs on colorectal cancer cells, have good anti-colorectal cancer effects, and significantly reduce tumor volume.

[0006] In order to achieve the above-mentioned object, the present invention provides a multifunctional liposome co-loaded with oxaliplatin and calcium peroxide nanoparticles. The liposome is composed of oxaliplatin, CaO2, lecithin, and ginsenoside Rg3, wherein the molar ratio of lecithin to ginsenoside Rg3 is 3-5:1; the molar ratio of lecithin to oxaliplatin is 15-20:1, the final concentration of CaO2 is 1-2 mg / mL, and the final concentration of OXA is 1-2 mg / mL.

[0007] The present invention provides a method for preparing multifunctional liposomes co-loaded with oxaliplatin and calcium peroxide nanoparticles, the method comprising the following steps:

[0008] Step 1: dissolving lecithin, ginsenoside Rg3 and CaO2 in a mixed solution of chloroform and anhydrous ethanol and rotary evaporating the mixture at 30-50° C. for 1-3 hours to form a transparent lipid film;

[0009] Step 2: Add oxaliplatin to a glucose solution, place on a shaker at 30-50°C for 1-4 hours, then sonicate at 100W for 10-20 minutes, and extrude the treated liposomes through a membrane with a pore size of 450 nm several times and through a membrane with a pore size of 220 nm several times;

[0010] The multifunctional liposomes co-loaded with oxaliplatin and calcium peroxide nanoparticles and their application of the present invention have the following advantages:

[0011] The multifunctional liposomes co-loaded with oxaliplatin and calcium peroxide nanoparticles, as well as their preparation and application, provide a combination of chemotherapeutic drugs and peroxide nanoparticles, improving the hypoxic microenvironment of solid tumors and enhancing the anti-tumor efficacy of oxaliplatin. The calcium peroxide nanoparticles provided herein have a particle size of less than 25.0 nm and a PDI value of less than 0.2, indicating that the calcium peroxide nanoparticles provided herein are uniformly distributed and exhibit no aggregation.

[0012] The multifunctional liposomes co-loaded with oxaliplatin and calcium peroxide nanoparticles provided by the present invention, as well as their preparation method and application, optimize conventional liposomes, introduce ginsenoside Rg3 to replace cholesterol to reduce the liposome particle size, and improve the targeting of the liposomes. At the same time, ginsenoside Rg3 is a good chemotherapy adjuvant, which can increase the anti-tumor effect and reduce the systemic toxicity of chemotherapy drugs when used in combination with chemotherapy drugs. Comparing the particle sizes of conventional cholesterol liposomes (Lip-OXA / CaO2) and ginsenoside Rg3 liposomes (Rg3-Lip-OXA / CaO2), the results show that the particle size of Lip-OXA / CaO2 is 121.54±5.63nm, and the particle size of Rg3-Lip-OXA / CaO2 is 104.78±3.96nm. In addition, the particle sizes of Lip-OXA / CaO2 and Rg3-Lip-OXA / CaO2 are both less than 0.2, indicating that the liposome particle size provided by the present invention is uniform and no coagulation occurs.

[0013] The multifunctional liposomes (Rg3-Lip-OXA / CaO2) co-loaded with oxaliplatin and calcium peroxide nanoparticles provided by the present invention have significant differences compared with the free drug (OXA), ordinary liposomes loaded with oxaliplatin (Lip-OXA), and ordinary liposomes co-loaded with oxaliplatin and calcium peroxide nanoparticles (Lip-OXA / CaO2). The Rg3-Lip-OXA / CaO2 of the present invention is more cytotoxic to HCT116 cells. Moreover, the Rg3-Lip-OXA / CaO2 of the present invention has the best inhibition rate on HCT116 cell tumors, with a significant reduction in tumor volume and a tumor inhibition rate of more than 80%. In terms of toxicity, the body weight of mice using the Rg3-Lip-OXA / CaO2 of the present invention increased, and the coefficients of major organs did not change significantly, indicating that the Rg3-Lip-OXA / CaO2 of the present invention has low toxicity.

[0014] Therefore, the Rg3-Lip-OXA / CaO2 of the present invention has a good tumor-suppressing effect in vivo and has relatively low toxicity. The preparation provided by the present invention has important theoretical value and good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Transmission electron microscope image of Example 1 of the present invention

[0016] Figure 2 This is the oxygen release curve of Example 1 of the present invention in water

[0017] Figure 3 Transmission electron microscope image of liposomes of Example 2 of the present invention

[0018] Figure 4 The particle size and PDI value changes of Example 2 of the present invention in different media.

[0019] Figure 5 The figures are the drug leakage rates of the liposomes of Example 2 of the present invention and Comparative Examples 1-3 at different temperatures.

[0020] Figure 6 The release curves of the drugs in the liposomes of Example 2 and Comparative Examples 1-3 in PBS are shown.

[0021] Figure 7 These are the fluorescence photos of intracellular uptake after HCT116 cells were incubated with different liposomes for a certain period of time.

[0022] Figure 8 The cell viability curves of the liposomes of Example 2 of the present invention and Comparative Examples 1-3 are shown.

[0023] Figure 9 The figures are the curves showing the changes in tumor volume and the tumor inhibition rates of tumor-bearing nude mice after treatment with the liposomes of Example 2 of the present invention and Comparative Examples 1-3.

[0024] Figure 10 The figure shows the curve of body weight change of tumor-bearing nude mice after treatment with the liposomes of Example 2 of the present invention and Comparative Examples 1-3. DETAILED DESCRIPTION

[0025] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0026] Example 1

[0027] A method for preparing calcium peroxide nanoparticles comprises:

[0028] 2 mL of a 0.2 g / mL CaCl₂ aqueous solution and 2 mL of a 1 mol / L ammonia solution were added to 80 mL of PEG₂O₂. After stirring, 2 mL of a 30% hydrogen peroxide solution was slowly added dropwise to the mixture, and stirring was continued at room temperature for 6 hours. The pH of the solution was adjusted to 11.5 with a 1 mol / L NaOH solution. The precipitate was collected by centrifugation at 9500 g for 10 minutes, washed twice with 0.1 mol / L sodium hydroxide solution, water, and anhydrous ethanol, respectively, to obtain white CaO₂ nanoparticles, which were then stored in anhydrous ethanol.

[0029] Example 2

[0030] A method for preparing multifunctional liposomes co-loaded with oxaliplatin and calcium peroxide nanoparticles, comprising:

[0031] Weigh 36 mg PC (egg yolk lecithin), 9 mg Rg3 (ginsenoside Rg3) and 1 mg CaO2 (calcium peroxide nanoparticles) into 5 mL chloroform and 5 mL anhydrous ethanol mixed solvent.

[0032] The above mixed solution is ultrasonically mixed for 5-10 min, and PC and Rg3 are fully dissolved in the organic solvent. After slow vacuum extraction at a rotation speed of 100-120 rpm, the solvent is rotary evaporated, the water bath temperature is 37°C, and a uniform transparent film is formed after 2 hours. Then 1 mL of oxaliplatin-containing glucose solution (the concentration of oxaliplatin is 1 mg / mL) is added, and the water is hydrated at 37°C for 2 h at a rotation speed of 100 rpm. Then, probe ultrasonic cooling for 3 s under an ice water bath for a total of 10 min, and extruding through a polycarbonate 80 nm membrane three times by a liposome extruder to obtain Rg3-Lip-OXA / CaO2.

[0033] Comparative Example 1

[0034] A preparation method of a single oxaliplatin ordinary liposome Lip-OXA, comprising:

[0035] Weigh 36 mg PC (egg yolk lecithin) and 4.5 mg Chol (cholesterol) into 5 mL chloroform and 5 mL anhydrous ethanol mixed solvent.

[0036] The above mixed solution is ultrasonically mixed for 5-10 min, and PC and Chol are fully dissolved in the organic solvent. After slow vacuum extraction at a rotation speed of 100-120 rpm, the solvent is rotary evaporated, the water bath temperature is 37°C, and a uniform transparent film is formed after 2 hours. Then 1 mL of oxaliplatin-containing glucose solution (the concentration of oxaliplatin is 1 mg / mL) is added, and the water is hydrated at 37°C for 2 h at a rotation speed of 100 rpm. Then, probe ultrasonic cooling for 3 s under an ice water bath for a total of 10 min, and extruding through a polycarbonate 80 nm membrane three times by a liposome extruder to obtain Lip-OXA.

[0037] Comparative Example 2

[0038] A preparation method of a single oxaliplatin ordinary liposome Lip-OXA, comprising:

[0039] Weigh 36 mg PC (egg yolk lecithin) and 9 mg Rg3 (ginsenoside Rg3) into 5 mL chloroform and 5 mL anhydrous ethanol mixed solvent.

[0040] The mixed solution is ultrasonically mixed for 5-10 minutes, PC and Rg3 are fully dissolved in the organic solvent, and then the solvent is rotary evaporated under slow vacuum at a rotation speed of 100-120 rpm. A uniform and transparent film is formed after 2 hours of water bath at 37°C. Then, 1 mL of oxaliplatin-containing glucose solution (the concentration of oxaliplatin is 1 mg / mL) is added, and the mixture is hydrated at 37°C under a rotation speed of 100 rpm for 2 hours. Then, the mixture is cooled for 3 seconds under an ice water bath for 10 minutes in total, and is extruded through a polycarbonate 80 nm membrane three times by using a liposome extruder to obtain Rg3-Lip-OXA.

[0041] Comparative Example 3

[0042] A preparation method of a common liposome Lip-OXA / CaO2 co-loading oxaliplatin and calcium peroxide nanoparticles, comprising:

[0043] 36 mg of PC (egg yolk lecithin), 4.5 mg of Chol (cholesterol), and 1 mg of CaO2 (calcium peroxide nanoparticles) are weighed and dissolved in 5 mL of chloroform and 5 mL of anhydrous ethanol mixed solvent.

[0044] The mixed solution is ultrasonically mixed for 5-10 minutes, PC and Chol are fully dissolved in the organic solvent, and then the solvent is rotary evaporated under slow vacuum at a rotation speed of 100-120 rpm. A uniform and transparent film is formed after 2 hours of water bath at 37°C. Then, 1 mL of oxaliplatin-containing glucose solution (the concentration of oxaliplatin is 1 mg / mL) is added, and the mixture is hydrated at 37°C under a rotation speed of 100 rpm for 2 hours. Then, the mixture is cooled for 3 seconds under an ice water bath for 10 minutes in total, and is extruded through a polycarbonate 80 nm membrane three times by using a liposome extruder to obtain Lip-OXA / CaO2.

[0045] The specific examples and comparative examples of the present application are as follows:

[0046] 1. The settings of the examples and comparative examples are as follows:

[0047] Table 1. Abbreviations of the examples and comparative examples of the present application

[0048]

[0049] 2. Morphological observation of calcium peroxide nanoparticles

[0050] The dried example 1 is uniformly dispersed in a methanol solution, an appropriate amount of sample is dropped on a copper mesh, dried, and then dyed with phosphotungstic acid for half an hour. The morphology of the calcium peroxide particles is observed by using a transmission electron microscope. The results are shown in Figure 1 The calcium peroxide nanoparticles are uniform spherical particles, and are uniformly dispersed without aggregation.

[0051] 3. Determination of particle size and PDI of calcium peroxide nanoparticles

[0052] The diluted Example 1 was taken and the particle size and PDI value (Polydispersity Index) were measured using a Malvern particle size analyzer at 25° C. The results are shown in Table 2.

[0053] Table 2 Particle size and PDI of Example 1 of the present invention (n=3)

[0054]

[0055] The particle size of calcium peroxide nanoparticles is 23.23±1.84 nm, which is less than 25 nm, and the PDI is 0.164±0.017, indicating that the particle size distribution of calcium peroxide nanoparticles is narrow.

[0056] 4. In vitro oxygen release from calcium peroxide nanoparticles

[0057] Take an appropriate amount of Example 1 and add it to PBS solutions of different pH values. Use a dissolved oxygen meter to measure the oxygen release capacity of Example 1 in water. The results are as follows: Figure 2 As shown in Figure 3, calcium peroxide nanoparticles release oxygen more quickly and completely in PBS with a pH of 5.5, indicating that the prepared calcium peroxide nanoparticles can stably and quickly release oxygen in an acidic environment.

[0058] 5. Determination of liposome encapsulation efficiency and drug loading

[0059] The liposomes of Example 2 and Comparative Examples 1-3 were demulsified, and the encapsulation efficiency (EE%) and drug loading (DL%) were determined by HPLC. The results are shown in Table 3.

[0060] Table 3 Encapsulation efficiency and drug loading of liposomes of Comparative Examples 1-3 and Example 2 of the present invention (n=3)

[0061]

[0062] The addition of calcium peroxide nanoparticles to the co-loaded liposomes did not affect the encapsulation efficiency of oxaliplatin. Rg3 as the membrane material had a relatively high encapsulation efficiency and drug loading capacity.

[0063] 6. Morphological observation of Rg3-Lip-OXA / CaO2 liposomes

[0064] Take the freshly prepared Example 2 and dilute it 100 times. Take an appropriate amount of sample and drop it on the copper mesh. After drying, stain it with phosphotungstic acid for half a minute and observe the morphology of liposomes using a transmission electron microscope. Figure 3 As shown, the liposomes are uniformly spherical, with a particle size of about 100-200 nm, and are evenly dispersed without aggregation.

[0065] 7. Determination of liposome particle size, PDI and Zeta potential

[0066] The diluted liposome solutions of Example 2 and Comparative Examples 1-3 were taken and the particle size, potential and PDI (Polydispersity Index) value were measured using a Malvern particle size analyzer at 25° C. The results are shown in Table 4.

[0067] Table 4 Particle size, PDI and Zeta potential of liposomes of Comparative Examples 1-3 and Example 2 of the present invention (n=3)

[0068]

[0069] As can be seen in Table 4, the multifunctional liposomes containing Rg3 have a smaller particle size than conventional liposomes. The PDI values ​​of each liposome are all less than 0.2, indicating uniform liposome distribution. The zeta potential values ​​of the liposomes are all negative, indicating that the liposomes are relatively stable and do not aggregate.

[0070] 8. Stability of Rg3-Lip-OXA / CaO2 liposomes

[0071] The liposomes of Example 2 were placed in three release media and the changes in particle size and PDI values ​​within 48 hours were measured. The three media were PBS solution, serum, and complete cell culture medium (10% FBS + 90% culture medium). PBS solution and serum are the most commonly used release media in in vitro release experiments, while complete cell culture medium is used to simulate the growth environment of human cells. In addition, the liposomes of Example 2 and Comparative Examples 1-3 were placed at 4°C and room temperature, respectively. The drug concentration encapsulated in the liposomes was measured at different time points (0h, 1h, 2h, 4h, 8h, 24h, 48h), recorded as Cn (n is a different time point), and the leakage percentage was substituted into the formula to obtain the leakage percentage, and a time-leakage percentage curve was plotted. The leakage rate was calculated as follows: leakage rate = (1-Cn / C0) × 100%.

[0072] like Figure 4 As shown, A is the particle size change of Rg3-Lip-OXA / CaO2 in Example 2 in different media, and B is the PDI value change of Rg3-Lip-OXA / CaO2 in Example 2 in different media. It can be seen that the particle size and PDI value both increase slightly with time, but are still within the appropriate range, and the PDI value is still less than 0.25, indicating that the liposome has good dispersibility.

[0073] like Figure 5As shown, the leakage rates of OXA of the liposomes of Example 2 of the present invention and Comparative Examples 1-3 at different temperatures (A: 4°C, B: room temperature) show that the leakage rate is low at lower temperatures, indicating that the liposomes are more stable and have a lower leakage rate at 4°C.

[0074] 9. In vitro release rate study

[0075] Liposomes from Example 2 and Comparative Examples 1-3 were placed in a dialysis bag and dialyzed at 37°C using PBS as the dialysis medium. Appropriate amounts were withdrawn at 0, 1, 2, 4, 8, 24, and 48 hours, and the OXA concentration, denoted as Cn (where n represents a different time point), was measured. A release rate curve was plotted with time as the abscissa and release rate as the ordinate. Release rate was calculated as: Release rate = (1-Cn / C0) × 100%.

[0076] like Figure 6 As shown in FIG1 , the release curves of OXA in PBS from the liposomes of Example 2 of the present invention and Comparative Examples 1-3 show that all liposomes exhibit slow release behavior, among which the slow release behavior of Rg3-Lip-OXA / CaO2 is the most obvious.

[0077] 10. In vitro targeting studies

[0078] To observe the uptake of different liposome preparations by HCT116 cells, two liposome preparations loaded with the fluorescent dye Rhodamine B (Rh B) were co-incubated with cells in an in vitro targeting experiment. The cells were then nuclear stained with Hoechst 33342 dye, and the changes in the cell uptake of liposomes were observed under a fluorescence microscope. Hoechst 33342 showed blue fluorescence under a fluorescence microscope, and Rhodamine B showed red fluorescence under a fluorescence microscope. Figure 7 The following are cell fluorescence images of free Rh B, Rh B-loaded liposomes (Lip-Rh B), and Rh B-loaded multifunctional liposomes (Rg3-Lip-Rh B). It can be seen that the fluorescence intensity of free Rh B in the cell is the weakest, while the fluorescence intensity of Lip-Rh B and Rg3-Lip-Rh B in the cell is stronger than that of free Rh B. Among them, the fluorescence intensity of Rg3-Lip-Rh B in the cell is the strongest. This is due to the targeting effect of Rg3 on the glucose transporter on the surface of HCT116 cells. This result shows that the Rg3 targeting fragment can specifically recognize and bind to the glucose transporter on the surface of HCT116 cells, enter the cell through receptor-mediated endocytosis, and increase the uptake of liposomes.

[0079] 11. In vitro cytotoxicity

[0080] HCT116 cells in the logarithmic growth phase (cultured in McCoy's 5A medium containing 10% fetal bovine serum and 1% streptomycin and penicillin mixed solution) were placed in a 25 cm 2 After digestion, the cells were centrifuged at 1000 rpm for 5 min and resuspended in the culture medium. 100 μL of the diluted cell suspension (2×10 3 Cells were inoculated into 96-well cell culture plates (cells / well). Control wells, blank wells, and drug-addition wells were set up at the same time. When the cells in the wells grew to 80%, drugs were added in triplicate, and different concentrations of drugs were added respectively. The cells were incubated for 48 hours. After the incubation, 10 μL of CCK-8 detection solution was added to each well. After incubation for another 1.5-2 hours, the cells were detected at a wavelength of 450 nm using a microplate reader. The relative cell survival rate was calculated, and the IC 50 .

[0081] like Figure 8 As shown, the cell viability curves of the liposomes of Example 2 of the present invention and Comparative Examples 1-3 show the in vitro cytotoxicity of different liposomes to HCT116, among which Rg3-Lip-OXA / CaO2 has the strongest cytotoxicity.

[0082] As shown in Table 5 below, the liposome IC of Example 2 of the present invention and Comparative Examples 1-3 are shown. 50 Among them, Rg3-Lip-OXA / CaO2 showed the strongest tumor cell killing ability and had the lowest IC 50 value.

[0083] Table 5 IC of Comparative Examples 1-3 and Example 2 of the present invention 50 value

[0084]

[0085] 12. In vivo anti-tumor effect and toxicity studies

[0086] Healthy female BALB / c nude mice weighing 18±2g were selected and raised in a clean environment at room temperature of 25°C with 12h day and night alternation. HCT116 cells were massively proliferated, and the cells were collected by centrifugation during the logarithmic growth phase. 0.1mL of cell suspension (5×106 cells / mouse) was subcutaneously injected into the flank of the nude mice. After the tumor volume grew to more than 80mm3, the mice were intravenously injected with the drug through the tail vein, once every four days, for a total of 6 times. The physical signs of the mice were observed, and the body weight and tumor volume of each group of mice were measured. After 30 days, the mice were killed, the tumor tissue was removed and weighed, and the tumor inhibition rate of each group was calculated. The dosage of Example 2 and Comparative Examples 1-3 was 5mg / kg OXA.

[0087] like Figure 9As shown, the curves of tumor volume changes and tumor inhibition rates of tumor-bearing nude mice after treatment with the liposomes of Example 2 of the present invention and Comparative Examples 1-3 are shown. Among them, the model group (Control) was injected with normal saline. It can be seen that the tumor volume of the Rg3-Lip-OXA / CaO2 group was significantly reduced, and the tumor inhibition rate was the highest.

[0088] like Figure 10 As shown, the weight change curves of tumor-bearing nude mice after treatment with the liposomes of Example 2 of the present invention and Comparative Examples 1-3 are shown. It can be seen that the weight changes of mice in each group are not significant, but the weight of mice in the Rg3-Lip-OXA / CaO2 group of the present invention has increased.

Claims

1. An oxaliplatin multifunctional liposome, characterized in that The liposomes are made of oxaliplatin, calcium peroxide nanoparticles, lecithin and ginsenoside Rg3.

2. The oxaliplatin multifunctional liposome according to claim 1, having a particle size of 104.78 ± 3.96 nm, a PDI value of 0.152 ± 0.011, a zeta potential of -29.67 ± 0.59, and an oxaliplatin encapsulation efficiency of 43.96 ± 3.18%.

3. A method for preparing oxaliplatin multifunctional liposomes, characterized in that: Lecithin and ginsenoside Rg3 were dissolved in a mixed solvent of chloroform and anhydrous ethanol and calcium peroxide nanoparticles were added. The mixture was rotary evaporated at 30-50°C for 1-3 hours on a rotary evaporator to form a transparent lipid film. Then, an aqueous solution of oxaliplatin in glucose was added to give a final oxaliplatin concentration of 1-2 mg / mL. The mixture was placed on a shaker at 30-50°C for 1-4 hours and then ultrasonically treated at 100 W for 10-20 minutes. The treated liposomes were extruded several times through a membrane with a pore size of 450 nm and several times through a membrane with a pore size of 220 nm to obtain the liposomes.

4. The method for preparing oxaliplatin multifunctional liposomes according to claim 3, wherein the method for preparing calcium peroxide nanoparticles comprises the following steps: adding 2 mL of 0.1 g / mL CaCl2 aqueous solution to 80 mL of PEG 200 After stirring evenly, 1 mL of 1 mol / L ammonia solution was added. Then, under rapid stirring, 1 mL of 30% hydrogen peroxide solution was slowly added to the mixture at a rate of 3 drops per minute. The mixture was stirred at room temperature for 6 hours. Subsequently, 1 mol / L NaOH solution was added under the action of ultrasound to adjust the pH value of the solution to 11.

5. The precipitate was collected by centrifugation at 9500g for 5 minutes and washed twice with 0.1 mol / L sodium hydroxide solution and ethanol respectively to obtain white calcium peroxide nanoparticles.

5. The method for preparing an oxaliplatin multifunctional liposome according to claim 3, wherein the molar ratio of lecithin to ginsenoside Rg3 is 3-5:1; the molar ratio of lecithin to oxaliplatin is 15-20:1, and the final concentration of CaO2 is 1-2 mg / mL.

6. Use of the multifunctional oxaliplatin liposomes according to claim 1 in the preparation of colorectal cancer drugs, characterized in that: The liposome is used as a medicine for preparing a medicine for treating colorectal cancer.

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