Capecitabine and baicalin nano polymer micelle and preparation method thereof

Abstract

The invention relates to the technical field of medicines, in particular to a nano polymeric micelle containing capecitabine and baicalin, the nano polymeric micelle has remarkable advantages, capecitabine, baicalin, vitamin E succinate and PLGA-PEG-streptavidin are prepared into the nano polymeric micelle, and the nano polymeric micelle is used for preparing the capecitabine, the baicalin, the vitamin E succinate and the PLGA-PEG-streptavidin. The prepared nanoemulsion has relatively small and uniform particle size, good stability, high drug loading capacity and encapsulation efficiency, and good long-term storage stability, shows a good anti-tumor effect through pharmacodynamic experiments of a mouse rectal cancer model, can significantly inhibit tumor growth, has relatively small liver and kidney toxicity, and is suitable for clinical application. The medicine application values of capecitabine and baicalin are enhanced.

Description

Technical Field

[0001] The present invention belongs to the technical field of preparations, and in particular relates to a composite preparation comprising capecitabine and baicalin and a preparation method thereof. Background Art

[0002] In recent years, the number of patients with colorectal cancer, especially rectal cancer, has increased significantly year by year. Since the early stage of the disease is often hidden, it is already in the late stage when it is discovered, which makes treatment more difficult. Although surgery is the main means of treatment, it has problems such as high recurrence rate and limited treatment effect. Therefore, drug treatment, especially the use of chemotherapy drugs, is particularly important.

[0003] Capecitabine is a commonly used oral anti-tumor drug. Since its launch, it has played an important role in the treatment of breast cancer and colorectal cancer. The existing preparations are mainly oral film-coated tablets, with specifications of 150mg and 500mg, which are characterized by rapid and complete absorption. However, capecitabine is prone to cause side effects such as hand-foot syndrome and bone marrow suppression, and the stability of ordinary tablets is poor. The surface of the tablets is prone to corrosion and disintegration during long-term storage. Capecitabine has strong cohesiveness, and the ester bond and sugar part in its structure are unstable and easily decomposed when exposed to water or alkali, so its long-term storage stability is not high.

[0004] CN104644601A discloses a capecitabine tablet, which is prepared by mixing capecitabine and a disintegrant, granulating and coating the mixture, and then compressing the mixture with a disintegrant, a filler and a lubricant. Compared with the prior art, the tablet has a fast dissolution rate and is basically not affected by humidity.

[0005] CN117442572A discloses a capecitabine solid dispersion tablet, which is obtained by tabletting 250 parts of capecitabine and 125-625 parts of a water-soluble dispersion carrier. The capecitabine solid dispersion tablet reaches a peak blood drug concentration 0.5 hours after oral administration.

[0006] CN114796147A discloses a capecitabine skeleton sustained-release preparation, which is prepared by combining capecitabine with a hydrophilic gel skeleton material to form a sustained-release preparation, which has a good sustained-release effect on capecitabine, can control the slow release of capecitabine, and maintains a stable blood drug concentration for 24 hours.

[0007] However, the above-mentioned new dosage forms all have high requirements on the preparation conditions of the preparations.

[0008] Baicalin is a flavonoid compound with anti-inflammatory, antioxidant and immunomodulatory effects. Studies have shown that baicalin can enhance the body's immune function and reduce the toxicity of chemotherapy drugs. Therefore, the combination of capecitabine and baicalin to prepare a new preparation is expected to provide a new idea for improving the treatment effect of rectal cancer, improving the side effects of capecitabine, and improving the stability of blood drug concentration. Summary of the invention

[0009] In view of the deficiencies in the prior art, the technical problem to be solved by the present invention is to provide a composite preparation of capecitabine and baicalin for treating rectal cancer or breast cancer.

[0010] The technical solution adopted by the present invention to solve the technical problem is: preparing a nano polymer micelle containing capecitabine and baicalin.

[0011] The first object of the present invention is to provide a nano polymer micelle containing capecitabine and baicalin. Specifically, the polymer micelle is prepared from capecitabine, baicalin, vitamin E succinate and PLGA-PEG-streptavidin.

[0012] Furthermore, the weight ratio of each component is as follows:

[0013]

[0014] Furthermore, the weight ratio of each component is as follows:

[0015]

[0016] The second object of the present invention is to provide a method for preparing the nano polymer micelles, specifically:

[0017] (1) mixing baicalin, PLGA-PEG-streptavidin and vitamin E succinate with an appropriate amount of organic solvent to form a mixed solution;

[0018] (2) removing the organic solvent by rotary evaporation of the mixed solution in step (1) to form a thin film;

[0019] (3) adding capecitabine to double distilled water, stirring to dissolve, and filtering to obtain a capecitabine solution;

[0020] (4) adding the film of step (2) into the capecitabine solution of step (3), stirring, filtering, and obtaining nano polymer micelles.

[0021] Preferably, the appropriate amount of organic solvent in step (1) is one of acetonitrile, dichloromethane or dimethyl sulfoxide and acetone.

[0022] Further preferably, the temperature of the rotary evaporation in step (2) is 55-70° C. and the time is 1-1.5 h.

[0023] More preferably, in step (3), filtration is performed using a 0.45 μm microporous filter membrane.

[0024] Further preferably, the specific operation of step (4) is: adding the film of step (2) to the capecitabine solution of step (3), stirring, filtering, and obtaining nano polymer micelles.

[0025] Specifically, the preparation method of the nano polymer micelle is:

[0026] (1) mixing baicalin, PLGA-PEG-streptavidin and vitamin E succinate with an appropriate amount of an organic solvent to form a mixed solution, wherein the organic solvent is selected from acetonitrile, dichloromethane or dimethyl sulfoxide, and acetone;

[0027] (2) removing the organic solvent by rotary evaporation of the mixed solution of step (1) at a temperature of 55 to 70° C. for 1 to 1.5 h to form a thin film;

[0028] (3) adding capecitabine to double distilled water, stirring to dissolve, and filtering with a 0.45 μm microporous filter membrane to obtain a capecitabine solution;

[0029] (4) adding the film of step (2) to the capecitabine solution of step (3), controlling the temperature at 40-60° C. and stirring at not less than 80,000 r / min, rinsing with ultrapure water to remove residual polymer and unencapsulated drug, and filtering through a 0.22 μm microporous filter membrane to obtain nanopolymer micelles.

[0030] The third object of the present invention is to provide the use of the nano polymer micelles, specifically: use of the nano polymer micelles in the preparation of anti-tumor drug preparations, preferably, the tumor is breast cancer or rectal cancer.

[0031] Furthermore, the anti-tumor drug preparation is an oral preparation, preferably one or more of nanoemulsion gel, soft capsule, tablet, capsule.

[0032] The present invention has the following beneficial effects:

[0033] (1) Capecitabine and baicalin were prepared into nanopolymer micelles, which significantly improved the dissolution rate of baicalin;

[0034] (2) The prepared nanoemulsion has a small and uniform particle size and good stability, which enhances the pharmaceutical application value of capecitabine and baicalin. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 is the particle size distribution diagram of the polymeric micelles of Example 2;

[0036] Figure 2 is the particle size distribution diagram of the polymeric micelles of Comparative Example 3;

[0037] Figure 3 The drug loading and encapsulation efficiency results of the capecitabine polymeric micelles prepared in Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, and Comparative Example 3 are shown.

[0038] Figure 4 Tumor volume and mass of mice in the experimental group, control group, and model group (compared with the model group, *P<0.05, **P<0.01);

[0039] Figure 5 Serum ALT and AST levels of mice in the experimental group, control group, and model group (compared with the model group, *P < 0.05, **P < 0.01);

[0040] Figure 6 The creatinine and urea nitrogen levels of mice in the experimental group, control group, and model group (compared with the model group, *P < 0.05, **P < 0.01); DETAILED DESCRIPTION

[0041] The present invention is further described below by way of examples. It should be correctly understood that the examples of the present invention are only used to illustrate the present invention, rather than to limit the present invention, and those skilled in the art should be aware that the examples described herein do not limit the present invention in any way.

[0042] Embodiment 1:

[0043] Group distribution ratio:

[0044] Preparation method:

[0045] (1) mixing baicalin, PLGA-PEG-streptavidin and vitamin E succinate with acetonitrile solvent to form a mixed solution;

[0046] (2) removing the organic solvent by rotary evaporation of the mixed solution of step (1) at a temperature of 60 to 65° C. for 1.5 h to form a thin film;

[0047] (3) adding capecitabine to double distilled water, stirring to dissolve, and filtering with a 0.45 μm microporous filter membrane to obtain a capecitabine solution;

[0048] (4) adding the film of step (2) to the capecitabine solution of step (3), controlling the temperature at 40-50° C. and stirring at 80,000 r / min, rinsing with ultrapure water to remove residual polymer and unencapsulated drugs, and filtering through a 0.22 μm microporous filter membrane to obtain nanopolymer micelles.

[0049] Embodiment 2:

[0050] Group distribution ratio:

[0051] Preparation method:

[0052] (1) mixing baicalin, PLGA-PEG-streptavidin and vitamin E succinate with an appropriate amount of an organic solvent to form a mixed solution, wherein the organic solvent is selected from acetonitrile, dichloromethane or dimethyl sulfoxide, and acetone;

[0053] (2) removing the organic solvent from the mixed solution of step (1) by rotary evaporation at a temperature of 65 to 70° C. for 1 h to form a thin film;

[0054] (3) adding capecitabine to double distilled water, stirring to dissolve, and filtering with a 0.45 μm microporous filter membrane to obtain a capecitabine solution;

[0055] (4) adding the film of step (2) to the capecitabine solution of step (3), controlling the temperature at 40-50° C. and stirring at 100,000 r / min, rinsing with ultrapure water to remove residual polymer and unencapsulated drugs, and filtering through a 0.22 μm microporous filter membrane to obtain nanopolymer micelles.

[0056] Example 3

[0057] Group distribution ratio:

[0058] The preparation method is the same as Example 1.

[0059] Example 4

[0060] Group distribution ratio:

[0061] The preparation method is the same as Example 2.

[0062] Example 5

[0063] Group distribution ratio:

[0064] The preparation method is the same as Example 1.

[0065] Comparative Example 1

[0066] Group distribution ratio:

[0067] (1) dissolving capecitabine, baicalin, polyoxyethylene hydrogenated castor oil and MPEG-2000-DSPE in an appropriate amount of organic solvent dichloromethane;

[0068] (2) placing the above solution in a rotary evaporator to evaporate the organic solvent to form a uniform thin film;

[0069] (3) Add an appropriate amount of water or buffer solution to the film and stir to hydrate it to form polymer micelles. Rinse with ultrapure water to remove residual polymer and unencapsulated drugs, and filter through a 0.22 μm microporous filter membrane to obtain nanopolymer micelles.

[0070] Compared with Example 2

[0071] Group distribution ratio: Capecitabine 100g

[0072] Baicalin 15g

[0073] Polysorbate-80 90g

[0074] Appropriate amount of organic solvent;

[0075] (1) Accurately weigh capecitabine and baicalin, add dichloromethane, and sonicate for 10 to 20 minutes to completely dissolve them;

[0076] (2) removing the solvent under reduced pressure and temperature control to obtain a dry film;

[0077] (3) adding distilled water to redissolve the mixture, ultrasonically hydrating the mixture to form polymeric micelles, washing the mixture with ultrapure water to remove residual polymer and unencapsulated drugs, and filtering the mixture through a 0.22 μm microporous filter membrane to obtain nano-polymer micelles.

[0078] Compared with Example 3

[0079] Group distribution ratio:

[0080] The preparation method is the same as Example 1.

[0081] Verification Example 1:

[0082] 1. Particle size detection:

[0083] Detection of the particle size of the capecitabine polymeric micelles prepared in Detection Example 2 and Comparative Example 3;

[0084] Test results such as Figure 1 , Figure 2 Displayed by Figure 1 , Figure 2 The results show that the particle size distribution of Example 2 is relatively uniform, with the largest distribution between 80-100 nm, while the particle size distribution of Comparative Example 3 is relatively dispersed.

[0085] 2. Calculation of drug loading and encapsulation efficiency:

[0086] The drug loading and encapsulation efficiency of the capecitabine polymeric micelles prepared in Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, and Comparative Example 3 were calculated;

[0087] Test results such as Figure 3 As shown by Figure 3 The results show that the encapsulation efficiency and drug loading of Example 1, Example 2 and Example 3 are significantly higher than those of Comparative Example 1, Comparative Example 2 and Comparative Example 3.

[0088] 3. Stability inspection

[0089] The tablets prepared in Examples 2-4 and Comparative Examples 1-3 were stored at 40°C±2°C and 75%±5% relative humidity for 6 months, and then their dissolution and impurity conditions were tested.

[0090] The dissolution was carried out according to the first method (basket method) of Part IV General Rules 0931 of the 2020 edition of the Chinese Pharmacopoeia, using 900 mL of 0.2% Tween 80 aqueous solution as the dissolution medium, the rotation speed was 100 rpm, the dissolution liquid temperature was (37±5°C), and the dissolution time was 10 minutes.

[0091]

[0092] Verification Example 2: Effect of polymeric micelles on rectal cancer mouse model

[0093] 1 Experimental Materials

[0094] 1.1 Experimental animals

[0095] Healthy BABL / c mice (provided by the Pharmacology Center of Lunan Pharmaceutical Group) weighed 20-22 g and began the experiment after 7 days of adaptive feeding.

[0096] 1.2 Experimental Reagents

[0097] Commercially available capecitabine tablets, the nanomicelles of Example 3 were further prepared according to conventional methods to obtain conventional tablets.

[0098] 2 Test methods

[0099] 2.1 Modeling

[0100] A mouse rectal cancer model was established by the AOM (azomethane) combined with DSS (dextran sulfate sodium) induction method. The specific operation was as follows: the mice were intraperitoneally injected with AOM (10 mg / kg), and one week later, they were given 3% DSS solution to drink freely for 7 days, and then normal drinking water was restored, and repeated for 3 cycles.

[0101] 2.2 Experimental drug administration

[0102] The rats with successful modeling were divided into three groups, namely the experimental group, the control group, and the model group, with 10 rats in each group. The experimental group was given the polymeric micelle preparation, 200 mg / kg of capecitabine, by gavage; the control group was given the commercially available common preparation, 400 mg / kg, by gavage; the model group was given the same volume of normal saline by gavage. Each group was given the drug by gavage for 14 consecutive days, followed by a 7-day rest, and then given the drug again for 14 consecutive days.

[0103] 3. Detection indicators and methods

[0104] 3.1 Tumor growth indicators

[0105] Tumor volume: At the end of the experiment, the tumor was dissected out and its long and short diameters were measured, and the tumor volume was calculated according to the following formula.

[0106] Tumor volume V = 0.5ab 2 , where (a is the major diameter and b is the minor diameter).

[0107] The tumors were removed and weighed, and the tumor inhibition rate was calculated according to the following formula.

[0108] Tumor inhibition rate % = (1-average tumor weight of drug administration group / average tumor weight of model group) × 100%

[0109] 3.2 Histopathological analysis

[0110] Tumor tissue: Tumor tissue was obtained for HE staining to observe the morphology and necrosis of tumor cells.

[0111] 3.3 Hematological indicators

[0112] At the end of the experiment, serum ALT, AST levels, creatinine and urea nitrogen levels were measured to evaluate liver toxicity and renal function.

[0113] 4 Statistical methods

[0114] SPSS 22.0 software was used to perform statistical analysis on the data, and P < 0.05 was considered statistically significant.

[0115] 5 Experimental results

[0116] 5.1 Daily observation of mice

[0117] During the experiment, the amount of exercise, diet, and drinking water of the two groups of mice were similar, but one mouse in the control group died. No abnormal pathological changes were found during the autopsy, and it is speculated that the death may have been caused by the large dose of the drug. At the same time, the weight change trend of the mice was also consistent, and the change was not significant (P>0.05). After the experiment, the tumor was removed for observation. Compared with the control group, the tumor volume and mass of the mice in the experimental group showed a significant decreasing trend (P<0.05). At the same time, according to the formula, the tumor inhibition rates of the experimental group and the control group were calculated to be 81.23% and 74.01%, respectively. The specific results are as follows Figure 4 shown.

[0118] 5.2 Histopathological observation of mice

[0119] HE staining results showed that the tumor cells in both groups of mice were arranged in disorder and the cell nuclei were fragmented, which was different from the model group.

Claims

1. A nanopolymer micelle comprising capecitabine and baicalin, characterized in that: The polymer micelle is prepared from capecitabine, baicalin, vitamin E succinate and PLGA-PEG-streptavidin.

2. The nano polymer micelle according to claim 1, characterized in that The weight ratio of each component is as follows:

3. The nano polymer micelle according to claim 1, characterized in that The preparation method of the nano polymer micelles comprises the following steps: (1) mixing baicalin, PLGA-PEG-streptavidin and vitamin E succinate with an appropriate amount of organic solvent to form a mixed solution; (2) removing the organic solvent by rotary evaporation of the mixed solution in step (1) to form a thin film; (3) adding capecitabine to double distilled water, stirring to dissolve, and filtering to obtain a capecitabine solution; (4) adding the film of step (2) to the capecitabine solution of step (3), stirring, filtering, and obtaining nano polymer micelles.

4. The nano polymer micelle according to claim 3, characterized in that In the step (1), the appropriate amount of organic solvent is one of acetonitrile, dichloromethane or dimethyl sulfoxide and acetone.

5. The nano polymer micelle according to claim 3, characterized in that The temperature of the rotary evaporation in the step (2) is 55 to 70° C. and the time is 1 to 1.5 h.

6. The nano polymer micelle according to claim 3, characterized in that In the step (3), filtration is performed using a 0.45 μm microporous filter membrane.

7. The nano polymer micelle according to claim 3, characterized in that The specific operation of step (4) is: adding the film of step (2) into the capecitabine solution of step (3), stirring, washing with water, filtering, and obtaining nano polymer micelles.

8. The nano polymer micelle according to claim 1, characterized in that The application of the nano polymer micelle in the preparation of anti-tumor drug preparations.

9. The nano polymer micelle according to claim 8, characterized in that The preparation is an oral preparation.

10. The nano polymer micelle according to claim 8, characterized in that The preparation is one or more of nanoemulsion gel, soft capsule, tablet and capsule.

Images