Silybin mixed micelle injection as well as preparation method and application thereof

Abstract

The invention provides a silibinin mixed micelle injection as well as a preparation method and application thereof, and belongs to the technical field of pharmaceutical preparations. The cholate / lecithin mixed micelle system constructed by adopting specific cholic acid and phospholipid has good stability, can be well compatible with the silibinin so as to well encapsulate the silibinin, can obviously improve the solubility of the silibinin, has higher safety under the condition of ensuring the stability of the silibinin, and can be used for preparing a medicine for treating the silibinin. Therefore, the bioavailability of the silibinin is greatly improved. According to the invention, a shearing dispersion method is adopted to prepare the mixed micelles, and the mixed micelles with high drug loading capacity, clarification and good stability for silibinin are formed by destroying and shearing the raw and auxiliary materials in a molecular level, so that the mixed micelles have better physical and chemical stability, the drug quality is easier to control, the process is more simplified, the clinical medication is safer and the cost is lower; and the accessibility of the medicine is improved.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical preparations, and in particular to a silybin mixed micelle injection, a preparation method and an application thereof. Background Art

[0002] Most patients with liver disease also have gallbladder disease. The medication cycle is long, it is difficult to be completely cured, and there are many cases of recurrence. Silibinin (SL) is a flavonoid compound extracted and refined from the seeds of the Asteraceae plant, Silybum marianum. 25 H 22 O 10 ), whose chemical structure is as follows:

[0003]

[0004] It is reported that silybin and its isomers have a stabilizing effect on liver cell membranes, and can prevent or avoid the loss of dissolved cell components (such as transaminase). Silybin can limit the penetration of certain hepatotoxic substances (such as α-amanitin) into the cell interior, enhance protein synthesis ability, enhance the repair and regeneration ability of liver cells, and is widely used in the treatment of acute and chronic hepatitis, early cirrhosis, liver poisoning and other diseases. However, silybin and its isomers have poor water solubility, low oral bioavailability (standard extracts containing 70-80% silybin can only absorb 20-30% from the gastrointestinal tract of animals and humans), and are metabolized quickly, so frequent administration is required to ensure that the blood drug concentration is within the effective range, and frequent administration is prone to accumulation poisoning. At the same time, the injection of silybin compounds has not yet appeared on the market, because its ordinary injection needs to add a large amount of auxiliary materials such as cosolvents or solubilizers, which will inevitably increase its toxicity and irritation, and increase the risk of clinical medication.

[0005] Chinese patent CN101439018A discloses an intravenous injection of silybin compound emulsion and its preparation method. Although the emulsion has the following advantages as a drug carrier: the safety of soybean oil as the oil phase and lecithin as the emulsifier to the body has been confirmed; soybean oil and lecithin as injection excipients have been included in the pharmacopoeias of my country and many other countries in the world, the dispersion of droplets in the emulsion is very large, and the bioavailability is high; the droplets in the emulsion can play a role in controlling diffusion, which can make the drug sustained-release and prolong the half-life; it can improve the targeting of the drug to the target tissue, especially with strong liver passive targeting, and can achieve high efficiency and low toxicity. However, there are two key problems to be solved when using emulsion as a drug carrier: 1) Whether the drug can be prepared into an emulsion depends on the physicochemical properties of the drug itself and its clinical indications. Not all raw materials are suitable for preparation into emulsions, and no prescription and preparation process of any drug-loaded emulsion can be directly obtained by applying existing technologies. 2) Whether the drug can achieve the expected effect after being prepared into an emulsion cannot be simply inferred, and experiments must be conducted to verify it. Summary of the invention

[0006] The purpose of the present invention is to provide a silibinin mixed micelle injection and a preparation method and application thereof, which can improve the solubility and oral bioavailability of silibinin.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a silybin mixed micelle injection, the preparation raw materials of which include silybin, phospholipid, bile acid, sodium hydroxide and water for injection;

[0009] The phospholipids include soybean lecithin and / or synthetic phospholipids;

[0010] The bile acid comprises one or two of glycocholic acid, deoxycholic acid and hyocholic acid;

[0011] The mass ratio of the phospholipid to silybin is 3 to 15:1;

[0012] The mass ratio of bile acid to silybin is 2 to 12:1;

[0013] The mass ratio of sodium hydroxide to silybin is 0.1 to 1:1;

[0014] The silybin content in the silybin mixed micelle injection is ≥98wt%;

[0015] The preparation method of the silybin mixed micelle injection comprises the following steps:

[0016] Under nitrogen protection, sodium hydroxide, water for injection and bile acid are mixed to carry out a neutralization reaction to obtain a sodium cholate solution;

[0017] The sodium cholate solution is mixed with phospholipids for emulsification to obtain a mixed micelle solution;

[0018] The mixed micelle solution is mixed with silybin and subjected to shear dispersion to obtain a silybin mixed micelle solution;

[0019] The pH value of the silybin mixed micelle solution is adjusted to 5-8 by using a pH regulator, and then sterilized to obtain a silybin mixed micelle injection.

[0020] Preferably, the mass ratio of bile acid to silybin is 5 to 10:1.

[0021] Preferably, the mass ratio of the phospholipid to silybin is 5 to 12:1.

[0022] Preferably, the mass ratio of sodium hydroxide to silybin is 0.2-0.8:1.

[0023] The present invention provides a method for preparing the silybin mixed micelle injection described in the above technical solution, comprising the following steps:

[0024] Under nitrogen protection, sodium hydroxide, water for injection and bile acid are mixed to carry out a neutralization reaction to obtain a sodium cholate solution;

[0025] The sodium cholate solution is mixed with phospholipids for emulsification to obtain a mixed micelle solution;

[0026] The mixed micelle solution is mixed with silybin and subjected to shear dispersion to obtain a silybin mixed micelle solution;

[0027] The pH value of the silybin mixed micelle solution is adjusted to 5-8, and sterilized to obtain a silybin mixed micelle injection.

[0028] Preferably, the neutralization reaction is carried out at a temperature of 50-60° C. and for 10 minutes.

[0029] Preferably, the emulsification temperature is 60-70° C., the time is 60 min, and the emulsification is carried out under high-speed shearing conditions, and the rotation speed of the high-speed shearing is 8000 rpm.

[0030] Preferably, the shear dispersion has a rotation speed of 8000 rpm, a temperature of 60-70° C., and a time of 90 min.

[0031] Preferably, the pH adjuster is a hydrochloric acid solution.

[0032] The present invention provides the use of the silibinin mixed micelle injection described in the above technical solution or the silibinin mixed micelle injection prepared by the preparation method described in the above technical solution in preparing silibinin drugs.

[0033] The silybin mixed micelle injection provided by the present invention adopts a bile salt / lecithin mixed micelle system constructed by specific bile acid and phospholipids, and the solubilization function is enhanced after the two are compounded, and the system has good stability, and the system can be well compatible with silybin so as to well encapsulate the silybin, and the solubility of silybin can be significantly improved. The system has higher safety under the condition of ensuring the stability of silybin, and thus the bioavailability of silybin is greatly improved.

[0034] The silybin mixed micelle injection of the present invention is a clear liquid with a light transmittance of not less than 85%, does not use highly toxic cosolvents such as polysorbate-80, and can effectively reduce adverse reactions of the drug in clinical use.

[0035] The present invention adopts a shear dispersion method to prepare mixed micelles, and forms mixed micelles with high drug loading, clarity and good stability for silybin by destroying and shearing the raw materials and auxiliary materials at the molecular level, so that the injection has better physical and chemical stability, makes the drug quality easier to control, makes clinical drug use safer, reduces the cost, and improves the accessibility of the drug.

[0036] The preparation method of the invention has simple process, is suitable for industrial production, can reduce costs, improve the accessibility of drugs, and has guiding significance for industrial application. DETAILED DESCRIPTION

[0037] The present invention provides a silybin mixed micelle injection, the preparation raw materials of which include silybin, phospholipid, bile acid, sodium hydroxide and water for injection;

[0038] The phospholipids include soybean lecithin and / or synthetic phospholipids;

[0039] The bile acid comprises one or two of glycocholic acid, deoxycholic acid and hyocholic acid;

[0040] The mass ratio of the phospholipid to silybin is 3 to 15:1;

[0041] The mass ratio of bile acid to silybin is 2 to 12:1;

[0042] The mass ratio of sodium hydroxide to silybin is 0.1 to 1:1;

[0043] The silybin content in the silybin mixed micelle injection is ≥98wt%;

[0044] The preparation method of the silybin mixed micelle injection comprises the following steps:

[0045] Under nitrogen protection, sodium hydroxide, water for injection and bile acid are mixed to carry out a neutralization reaction to obtain a sodium cholate solution;

[0046] The sodium cholate solution is mixed with phospholipids for emulsification to obtain a mixed micelle solution;

[0047] The mixed micelle solution is mixed with silybin and subjected to shear dispersion to obtain a silybin mixed micelle solution;

[0048] The pH value of the silybin mixed micelle solution is adjusted to 5-8 by using a pH regulator, and then sterilized to obtain a silybin mixed micelle injection.

[0049] In the present invention, the bile acid includes one or two of glycocholic acid, deoxycholic acid and hyocholic acid, preferably glycocholic acid; when the bile acid is two of the above bile acids, the present invention has no special limitation on the ratio of the two bile acids, and any ratio is acceptable.

[0050] In the present invention, the mass ratio of bile acid to silybin is 2 to 12:1, preferably 5 to 10:1, and more preferably 6 to 8:1.

[0051] In the present invention, the phospholipids include soybean lecithin and / or synthetic phospholipids, preferably soybean lecithin; when the phospholipids are soybean lecithin and synthetic phospholipids, the present invention has no particular limitation on the ratio of soybean lecithin and synthetic phospholipids, and any ratio is acceptable. In the present invention, the synthetic phospholipids are preferably DSPC (hydrogenated soybean lecithin).

[0052] In the present invention, the mass ratio of the phospholipid to silybin is 3 to 15:1, preferably 5 to 12:1, and more preferably 8 to 10:1.

[0053] In the present invention, the mass ratio of sodium hydroxide to silybin is 0.1 to 1:1, preferably 0.2 to 0.8:1, and more preferably 0.25 to 0.5:1.

[0054] In the present invention, the amount of the water for injection is preferably such that the content of silibinin in the silibinin mixed micelle injection is ≥98wt%, and the drug loading (the concentration of silibinin in the injection) of the silibinin mixed micelle injection is 1-10 mg / mL, more preferably 5 mg / mL.

[0055] The silybin mixed micelle injection of the present invention is a light yellow clear liquid, indicating that silybin can be 100% encapsulated.

[0056] The present invention provides a method for preparing the silybin mixed micelle injection described in the above technical solution, comprising the following steps:

[0057] Under nitrogen protection, sodium hydroxide, water for injection and bile acid are mixed to carry out a neutralization reaction to obtain a sodium cholate solution;

[0058] The sodium cholate solution is mixed with phospholipids for emulsification to obtain a mixed micelle solution;

[0059] The mixed micelle solution is mixed with silybin and subjected to shear dispersion to obtain a silybin mixed micelle solution;

[0060] The pH value of the silybin mixed micelle solution is adjusted to 5-8 by using a pH regulator, and then sterilized to obtain a silybin mixed micelle injection.

[0061] All preparation steps of the silybin mixed micelle injection of the present invention are carried out under nitrogen protection conditions.

[0062] In the present invention, sodium hydroxide is preferably added to water for injection, heated to a neutralization reaction temperature under stirring conditions, cholic acid is added after dissolution, and neutralization is performed under stirring conditions to obtain a sodium cholate solution. The present invention has no particular limitation on the stirring, and the reaction can be smoothly carried out according to a process well known in the art.

[0063] In the present invention, the neutralization reaction temperature is preferably 50-60°C, more preferably 50-55°C; and the time is preferably 10 minutes.

[0064] The present invention preferably heats to an emulsification temperature under high-speed shear conditions, adds phospholipids to a sodium cholate solution for emulsification; the emulsification temperature is preferably 60-70°C, more preferably 60-65°C; the time is preferably 60 min, the emulsification is carried out under high-speed shear conditions, and the high-speed shear rotation speed is preferably 8000 rpm; the emulsification is preferably carried out in a high-shear dispersing emulsifier.

[0065] The present invention preferably adds silybin to the mixed micelle solution under high-speed shearing conditions, heating to a shear dispersion temperature; the shear dispersion speed is preferably 8000 rpm, the temperature is preferably 60-70° C., and the time is preferably 90 min.

[0066] In the present invention, the pH regulator is preferably a hydrochloric acid solution, and the mass concentration of the hydrochloric acid solution is preferably 3wt%; the pH value of the silybin mixed micelle solution is preferably adjusted to 5.5-7.5, more preferably 6.5-7.0.

[0067] After adjusting the pH value to 5-8, the present invention preferably uses 0.22 μm filter membrane (PES) for filtration, nitrogen filling, controlling the residual oxygen content in the headspace of the bottle to ≤5%, and sterilization to obtain silybin mixed micelle injection.

[0068] The present invention provides the use of the silibinin mixed micelle injection described in the above technical solution or the silibinin mixed micelle injection prepared by the preparation method described in the above technical solution in the preparation of silibinin drugs. The present invention has no special limitation on the application method of the silibinin mixed micelle injection, and the application can be carried out according to methods well known in the art.

[0069] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0070] Example 1

[0071] The silybin mixed micelle injection provided in this embodiment is prepared from the following ingredients:

[0072] Silybin 10.0g, glycocholic acid 60.0g, soybean lecithin 80.0g, sodium hydroxide 5.0g, water for injection 845g;

[0073] The preparation process is carried out under nitrogen protection:

[0074] Weigh 5.0 g of sodium hydroxide and add it to 845 g of water for injection. Heat to 50°C under magnetic stirring. After the sodium hydroxide is dissolved, add 60.0 g of glycocholic acid and continue stirring for 10 min until it is completely dissolved to form a sodium glycocholate solution.

[0075] Turn on the high shear dispersing emulsifier, heat to 60°C under high-speed shear conditions of 8000 rpm, add 80.0 g of phospholipids into the glycocholate solution, and evenly disperse for 60 minutes until dissolved to form a mixed micelle solution;

[0076] Continue to add 10.0 g of silybin to the mixed micelle solution under high-speed shearing conditions of 8000 rpm and maintain the temperature at 60°C, and fully disperse for 90 minutes until dissolved to form a silybin mixed micelle solution;

[0077] After adjusting the pH value of the silybin mixed micelle solution to 5.5 with a 3wt% hydrochloric acid solution, the obtained mixed micelles were filtered through a 0.22μm filter membrane (PES), filled with nitrogen, and the residual oxygen content in the headspace of the bottle was controlled to be ≤5%, and sterilized to obtain a silybin mixed micelle injection with a drug loading of 10mg / mL and a silybin content of 99.0wt%.

[0078] Example 2

[0079] The silybin mixed micelle injection provided in this embodiment is prepared from the following ingredients:

[0080] Silybin 1.0g, glycocholic acid 6.0g, soybean lecithin 10.0g, sodium hydroxide 0.5g, water for injection 982.5g;

[0081] The preparation process is carried out under nitrogen protection:

[0082] Weigh 0.5 g of sodium hydroxide and add it to 982.5 g of water for injection. Heat to 50°C under magnetic stirring. After the sodium hydroxide is dissolved, add 6.0 g of glycocholic acid and continue stirring for 10 min until it is completely dissolved to form a glycocholic acid solution.

[0083] Turn on the high shear dispersing emulsifier, heat to 60°C under high-speed shear conditions of 8000 rpm, add 10.0 g of phospholipids into the glycocholate solution, and evenly disperse for 60 minutes until dissolved to form a mixed micelle solution;

[0084] Continue to add 1.0 g of silybin to the mixed micelle solution under high-speed shearing conditions of 8000 rpm and maintain the temperature at 60°C, and fully disperse for 90 minutes until dissolved to form a silybin mixed micelle solution;

[0085] After adjusting the pH of the silybin mixed micelle solution to 6.5 with a 3wt% hydrochloric acid solution, the obtained mixed micelles were filtered through a 0.22μm filter membrane (PES), filled with nitrogen, and the residual oxygen content in the headspace of the bottle was controlled to be ≤5%, and sterilized to obtain a silybin mixed micelle injection with a drug loading of 1mg / mL.

[0086] Example 3

[0087] Silybin mixed micellar injection is made of the following ingredients:

[0088] Silybin 5.0g, glycocholic acid 40.0g, soybean lecithin 50.0g, sodium hydroxide 2.0g, water for injection 903.0g;

[0089] The preparation process is carried out under nitrogen protection:

[0090] Weigh 2.0 g of sodium hydroxide and add it to 903.0 g of water for injection. Heat to 50°C under magnetic stirring. After the sodium hydroxide is dissolved, add 40.0 g of glycocholic acid and continue stirring for 10 min until it is completely dissolved to form a glycocholic acid solution.

[0091] Turn on the high shear dispersing emulsifier, heat to 60°C under high-speed shear conditions of 8000 rpm, add 50.0 g of soybean lecithin to the glycocholate solution, and evenly disperse for 60 minutes until dissolved to form a mixed micelle solution;

[0092] Continue to add 5.0 g of silybin to the mixed micelle solution under high-speed shearing conditions of 8000 rpm and maintain the temperature at 60°C, and fully disperse for 90 minutes until dissolved to form a silybin mixed micelle solution;

[0093] After adjusting the pH of the silybin mixed micelle solution to 7.5 with a 3wt% hydrochloric acid solution, the obtained mixed micelles were filtered through a 0.22μm filter membrane (PES), filled with nitrogen, and the residual oxygen content in the headspace of the bottle was controlled to be ≤5%, and sterilized to obtain a silybin mixed micelle injection with a drug loading of 5mg / mL.

[0094] Test Case

[0095] 1) According to the general rule 0401 of the 2020 edition of the "Chinese Pharmacopoeia", the transmittance of the silybin mixed micelle injection prepared in Examples 1 to 3 was determined by Shimadzu UV2600 ultraviolet-visible spectrophotometer. The results showed that the injection in Example 1 was an amber clear liquid with a transmittance of 86.5%; the injection in Example 2 was a light amber clear liquid with a transmittance of 92.5%; the injection in Example 3 was an amber clear liquid with a transmittance of 89.2%.

[0096] 2) Stability test: The drug stability of the silybin mixed micelle injection provided by the present invention (the sample prepared in Example 1) was investigated by an accelerated test method. The prepared silybin preparation was placed in a stability test box at a temperature of 40°C ± 2°C, and samples were taken once at 0 months, 1 month, 2 months, 3 months and 6 months, and the properties, transmittance, pH value and content of the test sample were checked regularly. The results are shown in Table 1.

[0097] Table 1 Accelerated stability test results of silybin mixed micelle injection in Example 1

[0098] time Characteristics Light transmittance pH content 0 month Light yellow clear liquid, no crystallization or precipitation 86.5％ 5.5 99.0wt% January Light yellow clear liquid, no crystallization or precipitation 86.1％ 5.6 100.2wt% February Light yellow clear liquid, no crystallization or precipitation 87.3％ 5.5 99.6wt% March Light yellow clear liquid, no crystallization or precipitation 85.1％ 5.6 98.9wt% June Light yellow clear liquid, no crystallization or precipitation 86.7％ 5.4 99.8wt%

[0099] The results in Table 1 show that under the temperature of 40°C±2°C, the silybin mixed micelle injection provided by the present invention has no crystallization or precipitation in appearance for 6 months, indicating that the micelle system in the injection is well compatible with silybin, the properties are clear, and the transmittance, pH value and content determination results are relatively stable, which meets the drug quality control requirements.

[0100] The silybin mixed micelle injection provided by the invention has good stability at 40° C. for 6 months, and can therefore be stored at room temperature with a shelf life of not less than 1 year.

[0101] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A silybin mixed micelle injection, characterized in that: The preparation raw materials include silybin, phospholipids, bile acid, sodium hydroxide, pH regulator and water for injection; The phospholipids include soybean lecithin and / or synthetic phospholipids; The bile acid comprises one or two of glycocholic acid, deoxycholic acid and hyocholic acid; The mass ratio of the phospholipid to silybin is 3 to 15:1; The mass ratio of bile acid to silybin is 2 to 12:1; The mass ratio of sodium hydroxide to silybin is 0.1 to 1:1; The silybin content in the silybin mixed micelle injection is ≥98wt%; The preparation method of the silybin mixed micelle injection comprises the following steps: Under nitrogen protection, sodium hydroxide, water for injection and bile acid are mixed to carry out a neutralization reaction to obtain a sodium cholate solution; The sodium cholate solution is mixed with phospholipids for emulsification to obtain a mixed micelle solution; The mixed micelle solution is mixed with silybin and subjected to shear dispersion to obtain a silybin mixed micelle solution; The pH value of the silybin mixed micelle solution is adjusted to 5-8 by using a pH regulator, and then sterilized to obtain a silybin mixed micelle injection.

2. The silybin mixed micelle injection according to claim 1, characterized in that: The mass ratio of bile acid to silybin is 5 to 10:

1.

3. The silybin mixed micelle injection according to claim 1, characterized in that: The mass ratio of the phospholipid to silybin is 5 to 12:

1.

4. The silybin mixed micelle injection according to claim 1, characterized in that: The mass ratio of the sodium hydroxide to silybin is 0.2-0.8:

1.

5. The method for preparing the silybin mixed micelle injection according to any one of claims 1 to 4, characterized in that: The following steps are involved: Under nitrogen protection, sodium hydroxide, water for injection and bile acid are mixed to carry out a neutralization reaction to obtain a sodium cholate solution; The sodium cholate solution is mixed with phospholipids for emulsification to obtain a mixed micelle solution; The mixed micelle solution is mixed with silybin and subjected to shear dispersion to obtain a silybin mixed micelle solution; The pH value of the silybin mixed micelle solution is adjusted to 5-8 by using a pH regulator, and then sterilized to obtain a silybin mixed micelle injection.

6. The preparation method according to claim 5, characterized in that: The temperature of the neutralization reaction is 50-60° C. and the time is 10 minutes.

7. The preparation method according to claim 5, characterized in that: The emulsification temperature is 60-70° C., the time is 60 min, and the emulsification is carried out under high-speed shearing conditions, and the rotation speed of the high-speed shearing is 8000 rpm.

8. The preparation method according to claim 5, characterized in that: The shear dispersion has a rotation speed of 8000 rpm, a temperature of 60-70° C., and a time of 90 min.

9. The preparation method according to claim 5, characterized in that: The pH adjuster is a hydrochloric acid solution.

10. Use of the silibinin mixed micelle injection according to any one of claims 1 to 4 or the silibinin mixed micelle injection prepared by the preparation method according to any one of claims 5 to 9 in the preparation of silibinin drugs.

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