Nanoparticle containing ginkgo leaf extract and rivaroxaban, preparation method and application thereof

Abstract

The invention provides nano-particles containing ginkgo biloba extract and rivaroxaban and a preparation method and application of the nano-particles, the composition comprises the ginkgo biloba extract, the rivaroxaban, sodium alginate and transfer eggs, the ginkgo biloba total flavonol glycosides in the ginkgo biloba extract are larger than or equal to 35%, and the total lactones in the ginkgo biloba extract are larger than or equal to 12%. The prepared nano-particles containing the ginkgo leaf extract and the rivaroxaban are good in stability, high in disintegration speed and excellent in gastrointestinal tract release characteristic, the bioavailability of the ginkgo leaf extract and the rivaroxaban is greatly improved, and the medication enthusiasm of a patient is improved; and the formula and the process are simple, industrial production is easy, and high popularization and application value is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical manufacturing, and in particular to nanoparticles containing ginkgo leaf extract and rivaroxaban, a preparation method and application thereof. Background Art

[0002] Rivaroxaban is an oral anticoagulant, usually provided in tablet form. Depending on the specifications and dosage forms, it is used to treat and prevent deep vein thrombosis and prevent pulmonary embolism, and stroke prevention in patients with atrial fibrillation. Clinically, for certain indications, such as the treatment of deep vein thrombosis (DVT), patients may need to take rivaroxaban twice a day (15 mg each time) for the first three weeks, and then change to once a day (20 mg each time). The frequency of taking is adjusted from multiple times a day to once a day to make it easier for patients to take and reduce the possibility of forgetting.

[0003] The main effects of Ginkgo biloba extract include benefiting the heart and lungs, removing dampness and stopping diarrhea. It is often used in the clinic to treat hyperlipidemia and cardiovascular and cerebrovascular diseases, such as myocardial ischemia, arrhythmia, cerebral thrombosis, etc. It has been recommended in many expert guidelines and consensus, including the "Guidelines for the Prevention and Treatment of Cerebrovascular Diseases in China" and the "MIMS Guidelines for the Use of Cardiovascular Diseases 2018 / 2019". In general, the dosage of Ginkgo biloba leaf is 2 tablets each time, 3 times a day. It is recommended to take it with or after meals to reduce gastrointestinal discomfort.

[0004] At present, the combination of ginkgo leaf extract and rivaroxaban is often considered in the treatment and prevention of cardiovascular diseases in clinic. However, since the frequency of use of the two drugs is different, patients often forget to take the medicine, which is very unfavorable to the cure of the disease. Therefore, considering the development of a new compound preparation to solve the problem of different frequency of use can greatly improve the compliance of patients with medication.

[0005] The solubility and permeability of drugs have an important impact on the pharmacological effects of drugs in the body. Rivaroxaban has low solubility and high permeability. Poor solubility directly affects its dissolution in the body, and ultimately affects its bioavailability. Ginkgo biloba extract is mainly flavonol glycosides and terpenoids, but ordinary preparations often have poor water solubility, precipitation during long-term storage, and darkening of color. In addition, when preparing drug preparations, drug stability also needs to be considered, including performance at different pH values.

[0006] In summary, the stability and functional properties of rivaroxaban and ginkgo leaf extract in preparation are affected by many factors. It is difficult to prepare a composite preparation with good stability and high solubility to prolong the medication time and improve patient compliance. Summary of the invention

[0007] In view of the shortcomings of the prior art, the present invention has found through research that after preparing ginkgo leaf extract and rivaroxaban into medicinal nanoparticles and then preparing them into conventional preparations, the dissolution and stability of the active ingredients are greatly improved, and the frequency of drug administration can be improved, solving the problem of patients forgetting to take their medicines.

[0008] The following is the technical solution adopted by the present invention.

[0009] One of the purposes of the present invention is to provide a medicinal nanoparticle containing ginkgo leaf extract and rivaroxaban, characterized in that the medicinal nanoparticle comprises ginkgo leaf extract, rivaroxaban, sodium alginate and ovotransferrin, wherein the ginkgo leaf extract is extracted from ginkgo leaves by heating with ethanol, and contains ≥60% of total ginkgo flavonol glycosides and ≥30% of total ginkgo lactones, and the weight ratio of the ginkgo leaf extract to rivaroxaban is 1:0.5-0.8.

[0010] The medicinal nanoparticles include the following raw materials in parts by weight:

[0011]

[0012] In a preferred embodiment, the medicinal nanoparticles include the following raw materials in parts by weight:

[0013]

[0014] Another object of the present invention is to provide a method for preparing the pharmaceutical nanoparticles comprising the following steps:

[0015] (1) dissolving ovotransferrin in pure water, controlling the temperature and stirring to obtain a protein aqueous solution;

[0016] (2) dissolving rivaroxaban in a small amount of dimethyl sulfoxide, mixing with the ginkgo leaf extract and the protein aqueous solution of step (1) and dissolving in an ethanol aqueous solution under stirring;

[0017] (3) slowly adding sodium alginate into the mixed solution of step (2) and stirring;

[0018] (4) removing the ethanol in step (3), freeze-drying, and obtaining composite nanoparticles.

[0019] In one embodiment, the mass concentration of the ovotransferrin aqueous solution in step (1) is 1.0-2.5% (w / w).

[0020] In one embodiment, the stirring temperature in step (1) is 0-8°C, and the stirring time is 4-15h.

[0021] In one embodiment, the volume concentration of the ethanol aqueous solution in step (2) is 70-90%, and the volume ratio of the ovotransferrin aqueous solution in step (1) to the ethanol in step (2) is 10:(1-3).

[0022] In one embodiment, step (3) comprises a pH adjustment step, specifically adjusting the pH to 5.0-6.5.

[0023] In the step (4), the temperature of the ethanol rotary evaporation is 50-60° C., and the vacuum degree is 0.06-0.1 MPa.

[0024] The freeze-drying condition of step (4) is -30°C, and the freeze-drying time is 24-72h.

[0025] The third object of the present invention is to provide a pharmaceutical preparation comprising the above-mentioned medicinal nanoparticles, wherein the preparation is an oral preparation, specifically one of granules, tablets, capsules, and mixtures.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] (1) Combining Ginkgo biloba extract with rivaroxaban to improve patient compliance;

[0028] (2) The prepared nanoparticles have better stability and solubility, greatly improving the bioavailability of the drug and making the drug efficacy better;

[0029] (3) The preparation method is simple, easy to replicate, low-cost, and has great industrialization potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The particle size and PDI value of the nanoparticles in Examples 1-5 after drying.

[0031] Figure 2 The particle size and PDI value of the nanoparticles in Examples 1-5 after drying.

[0032] Figure 3 is the nanoparticle embedding rate of Examples 1-5.

[0033] Figure 4 The pH stability test results of the nanoparticles of Examples 1-5 are shown. DETAILED DESCRIPTION

[0034] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0035] Example 1 Preparation of Nanoparticles

[0036] Formula:

[0037]

[0038] Preparation method:

[0039] (1) dissolving ovotransferrin in pure water and stirring at 5° C. for 15 h to obtain an ovotransferrin aqueous solution with a mass concentration of 2.5% (w / w);

[0040] (2) dissolving rivaroxaban in a small amount of dimethyl sulfoxide, mixing with ginkgo leaf extract and the protein aqueous solution of step (1) and dissolving in a 70% ethanol aqueous solution, wherein the volume ratio of the ovotransferrin aqueous solution in step (1) to the ethanol in step (2) is 10:2;

[0041] (3) slowly adding sodium alginate to the mixed solution of step (2), while adding acetic acid to adjust the pH to 6.0 while stirring;

[0042] (4) removing the ethanol in step (3), freeze-drying for 24 h, rotary evaporation temperature of 55° C., and vacuum degree of 0.06-0.08 MPa to obtain composite nanoparticles.

[0043] Example 2 Preparation of Nanoparticles

[0044] Formula:

[0045]

[0046] Preparation method:

[0047] (1) dissolving ovotransferrin in pure water and stirring at 8° C. for 4 h to obtain an ovotransferrin aqueous solution with a mass concentration of 1.0% (w / w);

[0048] (2) dissolving rivaroxaban in a small amount of dimethyl sulfoxide, mixing with ginkgo leaf extract and the protein aqueous solution of step (1) and dissolving in a 90% ethanol aqueous solution, wherein the volume ratio of the ovotransferrin aqueous solution in step (1) to the ethanol in step (2) is 10:1;

[0049] (3) slowly adding sodium alginate to the mixed solution of step (2), while adding acetic acid to adjust the pH to 5.0 while stirring;

[0050] (4) removing the ethanol in step (3), freeze-drying for 24 h, rotary evaporation at a temperature of 50° C., and vacuum degree of 0.10 MPa to obtain composite nanoparticles.

[0051] Example 3 Preparation of Nanoparticles

[0052] Formula:

[0053]

[0054] Preparation method:

[0055] (1) dissolving ovotransferrin in pure water and stirring at 0° C. for 15 h to obtain an ovotransferrin aqueous solution with a mass concentration of 1.0% (w / w);

[0056] (2) dissolving rivaroxaban in a small amount of dimethyl sulfoxide, mixing with ginkgo leaf extract and the protein aqueous solution of step (1) and dissolving in a 90% ethanol aqueous solution, wherein the volume ratio of the ovotransferrin aqueous solution in step (1) to the ethanol in step (2) is 10:1;

[0057] (3) slowly adding sodium alginate to the mixed solution of step (2), while adding acetic acid to adjust the pH to 5.5 while stirring;

[0058] (4) removing the ethanol in step (3), freeze-drying for 48 h, rotary evaporation at 60° C., and vacuum at 0.06-0.08 MPa to obtain composite nanoparticles.

[0059] Example 4 Preparation of Nanoparticles

[0060] Formula:

[0061]

[0062] The preparation method is the same as that of Example 1.

[0063] Example 5 Preparation of Nanoparticles

[0064] Formula:

[0065]

[0066] The preparation method is the same as that of Example 1.

[0067] Example 6 Preparation of Nanoparticles

[0068] Formula:

[0069]

[0070] The preparation method is the same as that of Example 1.

[0071] Example 7 Preparation of Nanoparticles

[0072] Formula:

[0073]

[0074] The preparation method is the same as that of Example 1.

[0075] Comparative Example 1 Preparation of Nanoparticles

[0076] Formula:

[0077]

[0078]

[0079] The ovotransferrin was replaced with soy protein isolate, and the preparation method was the same as in Example 1.

[0080] Comparative Example 2 Preparation of Nanoparticles

[0081] Formula:

[0082] Ginkgo biloba extract 100g

[0083] Rivaroxaban 250g

[0084] Ovotransferrin 50g

[0085] Preparation method:

[0086] (1) dissolving ovotransferrin in pure water and stirring at 0° C. for 15 h to obtain an ovotransferrin aqueous solution with a mass concentration of 1.0% (w / w);

[0087] (2) dissolving rivaroxaban in a small amount of dimethyl sulfoxide, mixing with ginkgo leaf extract and the protein aqueous solution of step (1) and dissolving in a 90% ethanol aqueous solution, wherein the volume ratio of the ovotransferrin aqueous solution in step (1) to the ethanol in step (2) is 10:1;

[0088] (3) Slowly adding Tween 60 to the mixed solution of step (2), while adding acetic acid to adjust the pH to 5.5 while stirring;

[0089] (4) removing the ethanol in step (3), freeze-drying for 48 h, rotary evaporation at a temperature of 50° C., and vacuum degree of 0.06-0.08 MPa to obtain composite nanoparticles.

[0090] Verification Example 1: Study on embedding efficiency, solubility and dispersibility

[0091] The embedding efficiency of the policosanol nanoformulations prepared in Examples 1-5 and Comparative Example 1 was calculated, and the particle size, Zeta potential and PDI were measured.

[0092] The embedding rate calculation formula is as follows:

[0093] Embedding rate = (1-surface policosan amount / total policosan amount) × 100%

[0094] The results are as follows Figure 1-3 As shown. Figure 1 , Figure 2 These are the values ​​after drying and after re-dissolution, Figure 3It can be seen from the results in the figure that compared with Comparative Example 1, the nanoformulations prepared in Examples 1-5 have good embedding rates, solubility and dispersibility (PDI), and the differences are significant (P<0.05).

[0095] Verification Example 2: pH Stability Study

[0096] The nanoparticles prepared in Example 1 were dispersed in an appropriate amount of water, and the pH values ​​were adjusted to 3, 4, 5, 6, 7, 8, and 9 with 0.1 M NaOH or HCl, respectively. The particle size, PDI, and Zeta potential value were tested after 24 hours.

[0097] The results are as follows Figure 4 The particle size of the nanoparticles changes differently at different pH values. It can be seen from the results in the figure that compared with Comparative Example 1, the nanoparticles prepared in Examples 1-5 have good pH stability, and the difference is significant (P < 0.05).

[0098] Verification Example 3: Pharmacokinetic Study in Rats

[0099] 1.1 Experimental Animals:

[0100] Select healthy SD rats: female, 8 weeks old, weight 180±20g, and feed them for one week.

[0101] 1.2 Drugs and preparations:

[0102] Commercially available Xarelto, nanoformulation of Example 2.

[0103] 1.3 Administration

[0104] The rats in the Xarelto group were fasted but not watered for 12 hours before administration. Example 2: The dose of Xarelto to rats was 0.5 mg / kg. After administration, the rats were fed a normal diet.

[0105] 1.4 Detection

[0106] 12 hours after administration, blood was collected from the orbital venous plexus of rats, centrifuged at a relative centrifugal force of 4000 rcf for 10 minutes to separate the plasma for testing. The bioequivalence of bioavailability (AUC) and maximum blood drug concentration (Cmax) was calculated under fasting administration.

[0107]

[0108] The results in Table 1 show that the preparation of the present invention can achieve similar AUC values ​​as the commercially available Xarelto, and the blood drug concentration is released steadily.

Claims

1. A medicinal nanoparticle containing ginkgo biloba extract and rivaroxaban, characterized in that: The medicinal nanoparticles contain ginkgo leaf extract, rivaroxaban, sodium alginate and ovotransferrin, wherein the ginkgo leaf extract is extracted from ginkgo leaves by heating with ethanol, and contains ≥60% of total ginkgo flavonol glycosides and ≥30% of total ginkgo lactones, and the weight ratio of the ginkgo leaf extract to rivaroxaban is 1:0.5-0.

8.

2. The pharmaceutical nanoparticle according to claim 1, characterized in that: The medicinal nanoparticles include the following raw materials in parts by weight:

3. The pharmaceutical nanoparticles according to claim 1, characterized in that: The preparation method of the medicinal nanoparticles comprises the following steps: (1) dissolving ovotransferrin in pure water, controlling the temperature and stirring to obtain a protein aqueous solution; (2) dissolving rivaroxaban in a small amount of dimethyl sulfoxide, mixing with the ginkgo leaf extract and the protein aqueous solution of step (1) and dissolving in an ethanol aqueous solution under stirring; (3) slowly adding sodium alginate into the mixed solution of step (2) and stirring; (4) removing the ethanol in step (3), freeze-drying, and obtaining composite nanoparticles.

4. The method for preparing the medicinal nanoparticles according to claim 3, characterized in that: The mass concentration of the ovotransferrin aqueous solution in step (1) is 1.0-2.5% (w / w).

5. The method for preparing the medicinal nanoparticles according to claim 3, characterized in that: The stirring temperature in step (1) is 0-8°C and the stirring time is 4-15h.

6. The method for preparing the medicinal nanoparticles according to claim 3, characterized in that: The volume concentration of the ethanol aqueous solution in step (2) is 70-90%, and the volume ratio of the ovotransferrin aqueous solution in step (1) to the ethanol in step (2) is 10:(1-3).

7. The method for preparing the medicinal nanoparticles according to claim 3, characterized in that: The step (3) includes a pH adjustment step, specifically adjusting the pH to 5.0-6.

5.

8. The method for preparing medicinal nanoparticles according to claim 3, wherein the temperature of the ethanol rotary evaporation in step (4) is 50-60°C and the vacuum degree is 0.06-0.1 MPa.

9. The method for preparing medicinal nanoparticles according to claim 3, wherein the freeze-drying condition in step (4) is -30°C and the freeze-drying time is 24-72 hours.

10. A pharmaceutical preparation comprising the pharmaceutical nanoparticles according to claims 1-9, wherein the preparation is an oral preparation, specifically one of granules, tablets, capsules, and mixtures.

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