HK1222319B - A silybin injection and preparation method thereof - Google Patents

Description

Silybin injection and preparation method thereof

Technical Field

The present invention relates to the field of medical technology, and in particular to a silibinin injection and a preparation method thereof.

Background Art

Silybin is a flavonoid compound extracted from the seed coat of the milk thistle seed, a medicinal plant in the Asteraceae family. Flavonoid compounds have poor water and fat solubility. Its structural formula is as follows:

Silybin has a significant hepatoprotective effect and has varying degrees of protective and therapeutic effects on various types of liver damage caused by liver poisons such as carbon tetrachloride, thioacetamide, muscarine, phalloidin, and crotalurine. It also has a certain inhibitory effect on the increase in alanine aminotransferase caused by carbon tetrachloride. As an ideal liver damage repair drug, silybin is widely used clinically to treat liver diseases such as acute and chronic hepatitis, liver fibrosis, and early cirrhosis. Due to its high efficacy and low toxicity (Yu Lecheng, Gu Changhai. Research Progress on the Pharmacological Effects of Silybin [J]. Chinese Journal of Hospital Pharmacy, 2001, 21(8): 493-494.), silybin has attracted much attention since its discovery.

Currently, the domestically marketed preparations include silybin meglumine tablets (approval number: National Medicine Standard H32026233; manufacturer: Jiangsu Zhongxing Pharmaceutical Co., Ltd.; approval number: National Medicine Standard H33022182; manufacturer: Zhejiang Dongri Pharmaceutical Co., Ltd.) and silybin capsules (trade name: Shuilinjia; approval number: National Medicine Standard H20040299; manufacturer: Tianjin Tasly Pharmaceutical Co., Ltd.). Both preparations are oral preparations. However, due to the poor water and lipid solubility of silybin (Wang Hongchen. Study on the Solubility of Silymarin [J]. Pharmaceutical Bulletin, 1983, 19(12): 23-25.), its oral bioavailability is low and it has a large first-pass effect. It is reported that the absolute bioavailability of silibinin after oral administration is only 0.95% (Wu JW, et al. Analysis of silibinin in rat plasma and bile for hepatobiliary excretion and oral bioavailability application[J]. J Pharm Biomed Anal, 2007, 45(4): 635–641.), which seriously weakens the efficacy of silibinin oral preparations. Therefore, people intend to improve the bioavailability of silibinin to further enhance its efficacy.

Numerous medical researchers have attempted to improve the solubility and oral absorption of silibinin using techniques such as salt formation, solubilization, and solid dispersion, but these efforts have been unsuccessful. Furthermore, Tianjin Tasly Pharmaceutical Co., Ltd. has developed a phospholipid complex of silibinin (generic name: Silybin Capsules; trade name: Shuilinjia; approval number: National Medicine Standard H20040299), which is now commercially available. Studies have shown that its solubility and bioavailability are significantly improved compared to ordinary silybin oral preparations, with bioavailability increased by 3 to 5 times (Filburn CR, et al. Bioavailability of silybin-phosphatidylcholine complex in dogs [J]. J Vet Pharmccol Ther, 2007, 30 (2): 132-138. silybin-phosphatidylcholine complex [M]. Altern Med Rev, ID: Thorne Research, Inc. 2009, 14 (4): 385-390.). However, its bioavailability is still not ideal. Therefore, people are considering developing silybin into an injection to fundamentally solve the problem of low oral bioavailability.

Song Yunmei et al. (Song Yunmei et al. Preparation of Silybin Nanoemulsion and Pharmacokinetics in Rabbits [J]. Journal of China Pharmaceutical University, 2005, 36(5): 427.) developed Silybin Nanoemulsion. Due to the poor solubility of Silybin, a large amount of polyoxyethylene castor oil (Cremophor RH) was added to the formulation. This material has severe allergic reactions and hemolysis, and has poor safety. In addition, since Silybin is soluble in water under alkaline conditions, some researchers have developed Silybin Meglumine Injection. However, Silybin is extremely unstable under alkaline conditions, resulting in poor long-term storage stability of the injection and easy degradation. Therefore, some people have prepared Silybin Meglumine solution into freeze-dried powder injection. Although this can solve the problem of storage stability, the pH value of Silybin Meglumine solution is about 11, which causes large local irritation during injection and many adverse reactions. According to the clinical use of silibinin in recent years, silibinin has been proven to have definite therapeutic effects and extremely low toxicity. In addition, silibinin has extremely wide clinical applications. If a safe, injectable silibinin preparation can be developed to improve its bioavailability and give full play to its pharmacological effects, thereby greatly improving its efficacy, it will surely produce great economic and social benefits.

Cyclodextrin inclusion complexes are compounds that encapsulate drug molecules within the cavity structure of cyclodextrin molecules, thereby solving the solubility problem of poorly water-soluble drugs. However, as we know, traditional cyclodextrin materials, such as β-cyclodextrin and hydroxypropyl-β-cyclodextrin, have significant hemolytic or nephrotoxicity. Chinese patent application CN200410041364.0, entitled "A method for preparing a cyclodextrin inclusion complex of milk thistle extract and its pharmaceutical preparation," and publication number CN159351A, features the core technical feature of dissolving milk thistle extract and cyclodextrin in ethanol or isopropanol, then removing the ethanol or isopropanol by evaporation under reduced pressure, and then adding excipients to prepare an oral preparation or directly packaging it into sterile powder for injection. The types of cyclodextrins described in the claims and the specification are: β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, methyl-β-cyclodextrin, glucosyl-β-cyclodextrin, and sulfonic acid-β-cyclodextrin. These cyclodextrins are still traditional cyclodextrins with large toxic side effects, and it is impossible to obtain preparations that can be used for intravenous injection in the true sense. Therefore, the entire embodiment is mainly based on oral preparations, and no further matching research is conducted on the above-mentioned cyclodextrins. Experiments have shown that none of the cyclodextrin types specified in the patent can be used to prepare a stable silybin injection (see Example 1 of the present invention); Chinese patent application CN02125823.6, entitled "Silymarin injection containing cyclodextrin or its derivatives", and publication number CN1391894A, discloses a core technical feature of the invention by adding silymarin to an aqueous solution containing cyclodextrin, then adjusting the pH of the solution to alkaline with sodium hydroxide to dissolve the silybin, and then adjusting the pH to 6.0-6.5 with hydrochloric acid to obtain a cyclodextrin inclusion complex of silymarin, wherein the type of cyclodextrin is exactly the same as that in the above-mentioned patent (Chinese patent application CN200410041364.0). It was found through experimental verification that silybin is dissolved in the alkaline aqueous solution process of sodium hydroxide, and the solution color gradually deepens and finally becomes a yellow-green solution. By full wavelength scanning, except that the original 287.5nm place has a larger absorption, a larger absorption peak occurs again at 326.0nm place, indicating that sodium hydroxide has a significant destructive effect on the structure of silymarin. Therefore, there are certain problems with the quality of the silymarin cyclodextrin inclusion compound prepared by this patent application, which may affect safety and effectiveness. Meanwhile, the freeze-dried powder injection prepared by this embodiment cannot be fully redissolved after freeze-drying (see Example 33 of the present invention). In addition, the cyclodextrin species recorded in its claims and the specification are still the traditional cyclodextrins with large toxic and side effects, and there is a potential safety hazard of hemolysis or nephrotoxicity.

Sulfobutyl ether-β-cyclodextrin (SBE-β-CD) is an ionized β-cyclodextrin (β-CD) developed by Cydex Corporation in the United States. Compared with traditional cyclodextrin materials, it has the advantages of good water solubility, no nephrotoxicity, and no hemolytic activity. To this end, the present invention, combining the physicochemical properties of silibinin, pioneered the use of sulfobutyl ether-β-cyclodextrin (SBE-β-CD) to address the solubility problem of silibinin. Through extensive experimental research, a sulfobutyl ether-β-cyclodextrin inclusion complex of silibinin was successfully developed. This is a safe and stable silibinin injection that fundamentally addresses the low oral bioavailability of silibinin and provides a new formulation for clinical research and application.

Summary of the Invention

The purpose of the present invention is to develop a safe and stable silibinin injection. In view of the inherent physical and chemical properties of silibinin and the shortcomings of existing preparations, a matching formulation was optimized, thereby successfully developing a silibinin injection.

The present invention provides a silibinin injection, characterized by containing silibinin, sulfobutyl ether-β-cyclodextrin, and an organic solvent for injection. The organic solvent for injection is used to dissolve silibinin. Silibinin is prepared into a molecular solution, which is then mixed with an aqueous solution of sulfobutyl ether-β-cyclodextrin to encapsulate the silibinin molecules within the sulfobutyl ether-β-cyclodextrin molecules, thereby forming a sulfobutyl ether-β-cyclodextrin inclusion complex of silibinin. If the prescription components do not contain an organic solvent for injection, the silibinin inclusion complex cannot be obtained according to the conventional method for preparing a cyclodextrin inclusion complex (directly adding the drug powder to the aqueous cyclodextrin solution and stirring until clear) (see Example 2 of the present invention). Therefore, the combined use of an organic solvent for injection and sulfobutyl ether-β-cyclodextrin in the present invention is one of the necessary conditions for preparing the silibinin inclusion complex and is also a core feature of the present invention.

The silibinin injection provided by the present invention may further contain a cosolvent. The cosolvent, on the one hand, can further increase the inclusion rate of silibinin and, on the other hand, facilitates rapid and effective resolubilization of the sample after lyophilization. The cosolvent enhances the quality of the silibinin injection prepared by the present invention. Therefore, the cosolvent is also an important technical feature of the present invention (see Example 3 of the present invention).

The present invention provides a silibinin injection, which is characterized by being prepared from the following components (weight to volume ratio):

The present invention provides a silibinin injection, which is characterized by being prepared from the following components (weight to volume ratio):

The present invention provides a silibinin injection, which is characterized by being prepared from the following components (weight to volume ratio):

The percentages of the various components of the silibinin injection of the present invention are all weight-to-volume ratios, i.e., the content of the solute contained in a unit volume of liquid, for example, including but not limited to g/ml, kg/L, etc., and can be scaled up or down on a year-on-year basis without changing the ratio.

The silybin injection, wherein the organic solvent for injection is selected from one or more of polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400, polyethylene glycol 600, anhydrous ethanol, and propylene glycol; preferably polyethylene glycol 400.

The cosolvent is selected from one or more of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), distearoylphosphatidylethanolamine-polyethylene glycol 5000 (DSPE-MPEG5000), Tween 80, polyoxyethylene castor oil (EL35), and poloxamer 188; preferably distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000).

The freeze-dried support agent is selected from one or more of mannitol, lactose, dextran 20, dextran 40, dextran 70, sucrose, xylitol, sorbitol, and trehalose; preferably mannitol, lactose, and dextran 40.

The pH adjuster is selected from one or more of citric acid, hydrochloric acid, glacial acetic acid, phosphoric acid, disodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium citrate, trisodium citrate, and sodium acetate; the pH value is adjusted to 2-8, preferably to 2-7; more preferably to 2.5-6.0.

The silibinin injection of the present invention is characterized by being a silibinin injection solution.

The silybin injection of the present invention is characterized by being a silybin lyophilized powder injection.

The silibinin injection of the present invention is a colorless clear liquid in solution form, and a white solid in freeze-dried powder injection form.

The preparation method of the silibinin injection solution of the present invention is characterized by comprising the following steps: weighing a formula amount of silibinin and an organic solvent for injection, optionally containing a cosolvent, mixing, heating and stirring at 40-70 DEG C to dissolve, and obtaining an organic phase; weighing a formula amount of sulfobutyl ether-β-cyclodextrin, placing the mixture in an appropriate amount of water for injection, stirring and dissolving, and obtaining an aqueous phase; mixing the organic phase and the aqueous phase, mixing them evenly, and then diluting the volume to the full amount with water for injection to obtain a drug solution; adjusting the pH value of the drug solution to 2-8 with a pH regulator; adding 0.05%-0.5% of activated carbon for injection with a weight volume percentage, adsorbing for 15-60 minutes, and then filtering the solution through a 1μm titanium rod to remove carbon and a 0.22μm filter element for sterilization; and packaging and sealing to obtain the silibinin injection solution.

The preparation method of the silibinin injection solution of the present invention preferably comprises the following steps: weighing a formula amount of silibinin and an organic solvent for injection, optionally containing a cosolvent, mixing, heating and stirring at 50-60°C to dissolve, and obtaining an organic phase; weighing a formula amount of sulfobutyl ether-β-cyclodextrin, placing the mixture in an appropriate amount of water for injection, stirring and dissolving, and obtaining an aqueous phase; mixing the organic phase with the aqueous phase, mixing them evenly, and then diluting the volume to the full amount with water for injection to obtain a drug solution; adjusting the pH value of the drug solution to 2.5-6.0 with a pH regulator; adding 0.1%-0.3% by weight volume of activated carbon for injection, adsorbing for 30-45 minutes, and then filtering the liquid through a 1μm titanium rod to remove carbon and a 0.22μm filter element for sterilization; and packaging and sealing to obtain the silibinin injection solution.

The preparation method of silybin lyophilized powder for injection of the present invention is characterized by comprising the following steps: weighing a formula amount of silybin and an organic solvent for injection, optionally containing a cosolvent, mixing, heating and stirring at 40-70 DEG C to dissolve, and obtaining an organic phase; weighing a formula amount of sulfobutyl ether-β-cyclodextrin and a lyophilized proppant, placing the mixture in an appropriate amount of water for injection, stirring and dissolving, and obtaining an aqueous phase; mixing the organic phase with the aqueous phase, mixing them evenly, and then diluting the volume to the full amount with water for injection to obtain a drug solution; adjusting the pH value of the drug solution to 2-8 with a pH regulator; adding 0.05%-0.5% of activated carbon for injection in a weight volume percentage, adsorbing for 15-60 minutes, and then filtering the solution through a 1μm titanium rod to remove carbon and a 0.22μm filter element for sterilization; packaging, lyophilizing, and sealing to obtain the silybin lyophilized powder for injection.

The preparation method of the silybin lyophilized powder for injection of the present invention preferably comprises the following steps: weighing a formula amount of silybin and an organic solvent for injection, optionally containing a cosolvent, mixing, heating and stirring at 50-60°C to dissolve, and obtaining an organic phase; weighing a formula amount of sulfobutyl ether-β-cyclodextrin and a lyophilized proppant, placing the mixture in an appropriate amount of water for injection, stirring and dissolving, and obtaining an aqueous phase; mixing the organic phase with the aqueous phase, mixing them evenly, and then diluting the volume to the full amount with water for injection to obtain a drug solution; adjusting the pH value of the drug solution to 2.5-6.0 with a pH regulator; adding 0.1%-0.3% of activated carbon for injection with a weight volume percentage, adsorbing for 30-45 minutes, and then filtering the solution through a 1μm titanium rod to remove carbon and a 0.22μm filter element for sterilization; packaging, lyophilizing, and sealing to obtain the silybin lyophilized powder for injection.

The silibinin injection of the present invention is characterized by combining the inherent physicochemical properties of silibinin to optimize the prescription composition and dosage that match the silibinin, thereby successfully developing a silibinin injection. The silibinin injection has the characteristics of being safe, stable, and having a high drug loading capacity, fundamentally overcoming the shortcoming of low bioavailability of existing oral preparations of silibinin, and is expected to further maximize the pharmacological effects of silibinin, thereby laying a foundation for the clinical application and research of silibinin.

Description of the drawings:

Figure 1: Raw material control solution-UV scanning curve

Figure 2: Raw material control solution - UV scanning absorbance list

Figure 3: UV scanning curve of the test solution of the present invention

Figure 4: UV scanning absorbance table of the test solution of the present invention

Figure 5: Comparison of patented test solution-UV scanning curve

Figure 6: Comparison of patented test solution - UV scanning absorbance list

DETAILED DESCRIPTION

The present invention will now be described in detail with reference to the embodiments, but the implementation of the present invention is not limited thereto.

Example 1: Comparative Study of Cyclodextrin Types

In Chinese patent application CN200410041364.0, the types of cyclodextrins described are: β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, methyl-β-cyclodextrin, glucosyl-β-cyclodextrin and sulfonic acid-β-cyclodextrin. The cyclodextrin selected in the present invention is sulfobutyl ether-β-cyclodextrin, which is the specific inclusion material of the present invention. In the comparative test, silybin injections containing different types of cyclodextrins were prepared using the same prescription composition, prescription dosage and preparation process. The injections were placed at room temperature for one week, and the inclusion stability of the injections was observed over time, that is, the cyclodextrin types were screened using the time for drug precipitation as an evaluation indicator. (1) Experimental plan

20 g of silybin, 1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 200 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase. 50 g of each of the above cyclodextrins was weighed, placed in 1000 ml of water for injection, and stirred to dissolve to obtain aqueous solutions of different types of cyclodextrins as the aqueous phase (when prepared according to this ratio, β-cyclodextrin is not completely soluble in water and is therefore discarded). The organic phase and the aqueous phase were stirred and mixed to form an inclusion solution with a concentration of approximately 2 mg/ml, which was then sterile-filtered using a 0.22 μm filter cartridge. The filtrate was dispensed into 5 ml bottles to obtain silybin injections of six different cyclodextrin types.

The injection was placed at room temperature for one week, and the inclusion stability of the silibinin injection was observed over time, that is, the effects of different cyclodextrin types on the inclusion stability of silibinin were investigated.

(2) Experimental results

Table 1 Results of the stability study of silybin encapsulated by different cyclodextrins

Cyclodextrin types 1 day 2 days 3 days 4 days 5 days 7 days Hydroxypropyl-β-cyclodextrin clear liquid clear liquid Slightly turbid Obvious turbidity Obvious turbidity Precipitation Hydroxyethyl-β-cyclodextrin clear liquid clear liquid clear liquid Slightly turbid Obvious turbidity Precipitation Methyl-β-cyclodextrin clear liquid Slightly turbid Obvious turbidity Precipitation Precipitation Precipitation Glucosyl-β-cyclodextrin clear liquid clear liquid clear liquid Slightly turbid Obvious turbidity Precipitation Sulfonic acid-β-cyclodextrin clear liquid clear liquid clear liquid Slightly turbid Obvious turbidity Obvious turbidity Sulfobutyl ether-β-cyclodextrin clear liquid clear liquid clear liquid clear liquid clear liquid clear liquid

Result analysis:

Silibinin injections prepared using the same preparation process and the same dosage formula exhibit distinct room temperature stability. None of the cyclodextrins described in Chinese patent application CN200410041364.0 can produce stable silibinin inclusion complex solutions, maintaining stability for only 2-4 days. However, silibinin inclusion complex solutions prepared using sulfobutyl ether-β-cyclodextrin show no significant changes for at least a week and are, in fact, quite stable at room temperature. Extensive testing has confirmed that sulfobutyl ether-β-cyclodextrin is the optimal inclusion material for silibinin. Therefore, the present invention's use of sulfobutyl ether-β-cyclodextrin for compatible inclusion complexes with silibinin is the optimal choice.

Example 2 Effect of Organic Solvents for Injection on the Quality of Silybin Injection

(1) The effect of the absence of organic solvents for injection on the quality of silybin injection

100g, 200g, 300g, 400g, and 500g of sulfobutyl ether-β-cyclodextrin were weighed and dissolved in water for injection to 1000ml, respectively, to obtain aqueous solutions containing 10%, 20%, 30%, 40%, and 50% sulfobutyl ether-β-cyclodextrin. Then, 1g of silybin was weighed and added to the aqueous solutions containing different sulfobutyl ether-β-cyclodextrin contents. The solution was stirred and the state of the solution was observed over time. The inclusion of silybin in sulfobutyl ether-β-cyclodextrin in the absence of an organic solvent for injection was assessed using appearance and properties. The results are shown in Table 2.

Table 2 Effect of the absence of organic solvents for injection on the quality of silybin injection

Result analysis:

Silibinin was directly added to a sulfobutyl ether-β-cyclodextrin solution using conventional methods and stirred to dissolve. Results showed that when the sulfobutyl ether-β-cyclodextrin concentration ranged from 10% to 50% and the drug loading was only 1 mg/ml, the solution remained turbid after 24 hours of stirring, showing no signs of dissolution. This suggests that silibinin's unique physicochemical properties preclude its preparation as a cyclodextrin inclusion complex via direct addition.

(2) Effects of organic solvents for injection on the quality of silybin injection

100g, 200g, 300g, 400g, and 500g of sulfobutyl ether-β-cyclodextrin were weighed and dissolved in water for injection to 1000ml, respectively, to obtain aqueous solutions containing 10%, 20%, 30%, 40%, and 50% sulfobutyl ether-β-cyclodextrin. 6g of silybin and 120g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55°C to dissolve, obtaining an organic phase. 21g of each organic phase was injected into the aforementioned series of aqueous solutions containing varying sulfobutyl ether-β-cyclodextrin contents, stirred, and the state of the solution observed over time. Appearance and properties were used as indicators to assess the inclusion of silybin in sulfobutyl ether-β-cyclodextrin in the presence of an injectable organic solvent. The results are shown in Table 3.

Table 3 Effects of organic solvents for injection on the quality of silybin injection

Result analysis:

Silybin is dissolved in an organic solvent for injection, and then a sulfobutyl ether-β-cyclodextrin solution is injected to prepare an inclusion compound. The resulting liquid state has a significantly increased clarity compared to the severe turbidity when the organic solvent for injection is not present. As can be seen, the organic solvent for injection in the prescription can preferably encapsulate silybin in the sulfobutyl ether-β-cyclodextrin molecule, significantly improving the encapsulation effect. Although a very small amount of drug is not encapsulated in the liquid, this problem is solved by adding an appropriate amount of cosolvent (see Example 3). Therefore, the presence of an organic solvent for injection in the prescription is necessary to match the specific physicochemical properties of silybin, and is one of the important technical features of the present invention, otherwise it will be impossible to encapsulate.

Example 3: Effect of cosolvent on the preparation of silybin injection

The cosolvent in the formulation is used, on the one hand, to address the problem of incomplete inclusion complex preparation, namely the phenomenon of individual fine particles suspended in the liquid mentioned in Example 2; on the other hand, the addition of the cosolvent ensures good resolubility of the freeze-dried sample. Therefore, the cosolvent enhances the quality of the silibinin injection prepared by the present invention. A comparative experiment examined the effects of adding and not adding a cosolvent on the quality of silibinin injection, using the same formulation composition and dosage. The cosolvent, distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), was used as an example for illustration.

(1) Preparation method and results of silybin injection without cosolvent

Weigh 2g of silybin and 20g of polyethylene glycol 400, mix them, and heat with stirring at 55°C to dissolve them to obtain an organic phase. Weigh 50g of sulfobutyl ether-β-cyclodextrin and 50g of mannitol, dissolve them in water for injection to 1000ml, and stir to dissolve them to obtain an aqueous phase. The organic and aqueous phases are stirred to obtain an inclusion solution, which is sterile-filtered using a 0.22μm filter cartridge. Intermediate determination is performed by sampling and observing the solution state before and after filtration and the labeled percentage of the drug. The filtrate is dispensed into 5ml bottles, lyophilized, and sealed. After lyophilization, reconstitute the solution with water for injection, observe the solution state under a light detector, and pass the reconstituted solution through a 0.22μm filter membrane. The labeled percentage of the drug in the subsequent filtrate is determined. The results are shown in Table 4.

Table 4 Results of silybin injection without cosolvent

project Solution state Labeled content percentage Before inclusion liquid filtration Basically clear, with a few fine particles suspended 99.76% After filtration of the inclusion liquid Complete clarification 97.07% After freeze-dried reconstitution There are obvious suspended particles and opalescence 95.33%

Result analysis:

Without cosolvent, some drugs cannot be completely encapsulated during the preparation process, sterilization filtration is not smooth, resistance is large, drugs are retained, which is not conducive to batch production, and cannot be completely re-dissolved after freeze-drying.

(2) Preparation of inclusion solution containing cosolvent

2g of silybin, 0.1g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55°C to dissolve to obtain an organic phase. 50g of sulfobutyl ether-β-cyclodextrin and 50g of mannitol were weighed, dissolved in water for injection to 1000ml, and stirred to dissolve to obtain an aqueous phase. The organic and aqueous phases were stirred to obtain an inclusion solution, which was sterile-filtered using a 0.22μm filter cartridge. Intermediate determination was performed by sampling the solution before and after filtration and observing the labeled percentage of the drug. The filtrate was dispensed into 5ml bottles, lyophilized, and sealed. After lyophilization, water for injection was added for reconstitution. The solution was observed under a light detector. The reconstituted solution was filtered through a 0.22μm filter membrane, and the labeled percentage of the drug in the subsequent filtrate was determined. The results are shown in Table 5.

Table 5 Results of silybin injection containing cosolvent

project Solution state Labeled content percentage Before inclusion liquid filtration Complete clarification 100.04% After filtration of the inclusion liquid Complete clarification 99.92% After freeze-dried reconstitution Complete clarification 99.78%

Result analysis:

After adding the cosolvent, the drug was completely encapsulated during the preparation process, sterilization and filtration were smooth, the drug content did not change significantly before and after filtration, and it was completely resolubilized after lyophilization. Therefore, the addition of the cosolvent in the formulation is an important technical feature of the present invention.

Example 4 Silybin injection

2 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 60 g of mannitol were weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 5 Silybin injection

2 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin was weighed, placed in 900 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged and sealed to obtain silybin injection.

Example 6 Silybin injection

10 g of silybin, 10 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 100 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 60° C. to dissolve to obtain an organic phase; 300 g of sulfobutyl ether-β-cyclodextrin, 50 g of xylitol, 50 g of sorbitol, and 100 g of mannitol were weighed, placed in 350 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 8.00 with potassium dihydrogen phosphate and dipotassium hydrogen phosphate; 5 g of activated carbon for injection was added and adsorbed for 15 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 7 Silybin Injection

8 g of silybin, 5 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 50 g of polyethylene glycol 50 were weighed, mixed, and heated with stirring at 70° C. to dissolve to obtain an organic phase; 200 g of sulfobutyl ether-β-cyclodextrin, 50 g of sucrose, and 100 g of mannitol were weighed, placed in 550 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 7.00 with sodium dihydrogen phosphate and disodium hydrogen phosphate; 3 g of activated carbon for injection was added and adsorbed for 20 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 8 Silybin injection

5 g of silybin, 3 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 50° C. to dissolve to obtain an organic phase; 100 g of sulfobutyl ether-β-cyclodextrin, 40 g of trehalose, and 60 g of mannitol were weighed, placed in 700 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 2.00 with hydrochloric acid and citric acid; 2 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 9 Silybin Injection

0.5 g of silybin, 0.01 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 5 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 40° C. to dissolve to obtain an organic phase; 20 g of sulfobutyl ether-β-cyclodextrin was weighed, placed in 950 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 6.00 with phosphoric acid and dipotassium hydrogen phosphate; 0.5 g of activated carbon for injection was added and adsorbed for 60 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged and sealed to obtain silybin injection.

Example 10 Silybin injection

1 g of silybin and 10 g of polyethylene glycol 400 are weighed, mixed, and heated with stirring at 50° C. to dissolve to obtain an organic phase; 30 g of sulfobutyl ether-β-cyclodextrin and 40 g of mannitol are weighed, placed in 900 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly blended, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 2.50 with hydrochloric acid, citric acid, and disodium citrate; 1 g of activated carbon for injection is added and adsorbed for 40 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 11 Silybin injection

1 g of silybin, 0.05 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 10 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 20 g of sulfobutyl ether-β-cyclodextrin, 20 g of lactose, and 10 g of dextran 70 were weighed, placed in 900 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 5.00 with phosphoric acid and sodium dihydrogen phosphate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 12 Silybin injection

1 g of silybin, 0.2 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 10 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 20 g of sulfobutyl ether-β-cyclodextrin, 20 g of lactose, and 10 g of dextran 20 were weighed, placed in 900 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 4.50 with glacial acetic acid and sodium acetate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 13 Silybin injection

1 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of anhydrous ethanol were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 30 g of sulfobutyl ether-β-cyclodextrin and 40 g of mannitol were weighed, placed in 900 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid and trisodium citrate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 14 Silybin Injection

1 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of propylene glycol are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 30 g of sulfobutyl ether-β-cyclodextrin and 40 g of mannitol are weighed, placed in 900 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly blended, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 3.50 with citric acid and trisodium citrate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 15 Silybin injection

2 g of silybin, 1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 40 g of anhydrous ethanol were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 60 g of lactose were weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid and disodium hydrogen phosphate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 16 Silybin Injection

2 g of silybin, 1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 40 g of propylene glycol are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 60 g of lactose are weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly mixed, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 3.50 with citric acid and disodium hydrogen phosphate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 17 Silybin Injection

2 g of silybin, 0.2 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 50 g of polyethylene glycol 400 are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 50 g of sulfobutyl ether-β-cyclodextrin and 60 g of dextran 40 are weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly mixed, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 3.50 with citric acid and trisodium citrate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 18 Silybin Injection

2 g of silybin, 0.4 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 30 g of sulfobutyl ether-β-cyclodextrin, 30 g of dextran 40, and 20 g of lactose were weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.00 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and the solution was then filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 19 Silybin injection

2 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 5000 (DSPE-MPEG5000), and 30 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 60 g of mannitol were weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.00 with citric acid and trisodium citrate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 20 Silybin injection

2 g of silybin, 0.5 g of distearoylphosphatidylethanolamine-polyethylene glycol 5000 (DSPE-MPEG5000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 50 g of mannitol were weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 4.00 with citric acid and trisodium citrate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 21 Silybin injection

2 g of silybin, 10 g of Tween 80, and 20 g of polyethylene glycol 400 are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 70 g of mannitol are weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly blended, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 4.00 with citric acid and trisodium citrate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 22 Silybin injection

2 g of silybin, 10 g of polyoxyethylene castor oil, and 20 g of polyethylene glycol 400 are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 110 g of sulfobutyl ether-β-cyclodextrin is weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly blended, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 4.00 with citric acid and trisodium citrate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged and sealed to obtain silybin injection.

Example 23 Silybin Injection

2 g of silybin, 10 g of poloxamer 188, and 20 g of polyethylene glycol 400 are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin, 20 g of lactose, and 40 g of mannitol are weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly blended, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 5.00 with phosphoric acid and sodium dihydrogen phosphate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 24 Silybin Injection

2 g of silybin, 5 g of Tween 80, and 20 g of polyethylene glycol 400 are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 450 g of sulfobutyl ether-β-cyclodextrin is weighed, placed in 400 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly blended, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 5.00 with phosphoric acid and disodium hydrogen phosphate; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged and sealed to obtain silybin injection.

Example 25 Silybin injection

Weigh 2 g of silybin, 5 g of polyoxyethylene castor oil, 5 g of propylene glycol, and 15 g of anhydrous ethanol, mix them, and heat with stirring at 55° C. to dissolve them to obtain an organic phase; weigh 100 g of sulfobutyl ether-β-cyclodextrin, place it in 850 ml of water for injection, and stir to dissolve it to obtain an aqueous phase; mix the organic phase and the aqueous phase, mix them evenly, and then dilute the volume to 1000 ml with water for injection to obtain a drug solution; adjust the pH value of the drug solution to 3.50 with citric acid; add 1 g of activated carbon for injection and adsorb it for 30 minutes; then filter the solution through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element to sterilize it; package and seal the container to obtain silybin injection.

Example 26 Silybin Injection

2 g of silybin, 0.2 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 200 are weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 60 g of mannitol are weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase are mixed and uniformly mixed, and then the volume is adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution is adjusted to 3.00 with citric acid; 1 g of activated carbon for injection is added and adsorbed for 30 minutes, and then the solution is filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution is packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 27 Silybin Injection

2 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 300 were weighed, mixed, and heated with stirring at 60° C. to dissolve to obtain an organic phase; 40 g of sulfobutyl ether-β-cyclodextrin and 50 g of mannitol were weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid and sodium citrate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 28 Silybin injection

2 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 30 g of polyethylene glycol 600 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 50 g of sulfobutyl ether-β-cyclodextrin and 60 g of mannitol were weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 29 Silybin injection

2 g of silybin, 0.05 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 50 g of sulfobutyl ether-β-cyclodextrin, 30 g of lactose, and 30 g of mannitol were weighed, placed in 850 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid and trisodium citrate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 30 Silybin injection

3 g of silybin, 2 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 60° C. to dissolve to obtain an organic phase; 80 g of sulfobutyl ether-β-cyclodextrin, 40 g of dextran 40, and 60 g of mannitol were weighed, placed in 750 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 50 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 31 Silybin Injection

3 g of silybin, 3 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), 25 g of propylene glycol, and 5 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 50 g of sulfobutyl ether-β-cyclodextrin and 60 g of mannitol were weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 40 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 32 Silybin injection

3 g of silybin, 1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 30 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 60 g of sulfobutyl ether-β-cyclodextrin and 60 g of mannitol were weighed, placed in 800 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid and trisodium citrate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin freeze-dried powder injection.

Example 33 Silybin injection

4 g of silybin, 1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), 10 g of anhydrous ethanol, 10 g of propylene glycol, and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 80 g of sulfobutyl ether-β-cyclodextrin and 100 g of mannitol were weighed, placed in 750 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 6.00 with sodium dihydrogen phosphate and dipotassium hydrogen phosphate; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged, freeze-dried, and sealed to obtain silybin lyophilized powder injection.

Example 34 Silybin Injection

2 g of silybin, 0.1 g of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-MPEG2000), and 20 g of polyethylene glycol 400 were weighed, mixed, and heated with stirring at 55° C. to dissolve to obtain an organic phase; 500 g of sulfobutyl ether-β-cyclodextrin was weighed, placed in 400 ml of water for injection, and stirred to dissolve to obtain an aqueous phase; the organic phase and the aqueous phase were mixed and uniformly mixed, and then the volume was adjusted to 1000 ml with water for injection to obtain a drug solution; the pH value of the drug solution was adjusted to 3.50 with citric acid; 1 g of activated carbon for injection was added and adsorbed for 30 minutes, and then the solution was filtered through a 1 μm titanium rod to remove carbon and a 0.22 μm filter element for sterilization; the solution was packaged and sealed to obtain silybin injection.

Example 35 Investigation of Dilution Stability of Silybin Injection

Silibinin injections prepared in Examples 4, 6, 9, 13, 17, and 32 were prepared, and the lyophilized powder injections were reconstituted with physiological saline to the concentration of the solution before lyophilization. The silibinin injections and the reconstituted lyophilized powder injections were each diluted 10-fold with physiological saline to obtain dilutions. The dilutions were placed at room temperature, sampled over time, and filtered through a 0.45-μm filter membrane. The silibinin content in the subsequent filtrate was measured over time to investigate the dilution stability of the silibinin injections.

The chromatographic conditions were octadecylsilane bonded silica gel as the filler; methanol-water-glacial acetic acid (48:52:1) as the mobile phase; the column temperature was 40°C; and the detection wavelength was 288 nm.

Determination method: Accurately measure 1 ml of diluent and dilute it to 10 ml with mobile phase as the test sample; accurately measure 10 μl and inject it into the liquid chromatograph, and record the peak area; taking the peak area at 0 hours as 100%, calculate the change in peak area percentage over time. The results are shown in Table 6:

Table 6 Dilution stability test results of silybin injection

sample 0 hours 8 hours 12 hours 16 hours 20 hours 24 hours Example 4 100% 100.2% 100.4% 99.8% 99.6% 98.8% Example 6 100% 99.7% 100.1% 99.6% 99.2% 99.5% Example 9 100% 99.9% 99.7% 100.3% 100.0% 99.2% Example 13 100% 99.9% 100.2% 100.1% 99.8% 99.6% Example 17 100% 101.0% 100.3% 100.7% 99.7% 100.1% Example 32 100% 100.5% 99.6% 98.9% 99.7% 99.1%

The results show that the dilution stability of the silibinin injection of the present invention in physiological saline before clinical use is greater than 24 hours, which can fully meet the requirements of clinical use.

Example 36 Verification of the preparation method described in Chinese patent application CN02125823.6

Chinese patent application CN02125823.6, entitled "Silymarin Injection Containing Cyclodextrin or Its Derivatives," and publication number CN1391894A, describes a method for injecting silymarin into a cyclodextrin-containing aqueous solution. The solution's pH is then adjusted to alkaline with sodium hydroxide to dissolve silybin. Following dissolution, the pH is then adjusted to 6.0-6.5 with hydrochloric acid to obtain a cyclodextrin inclusion complex of silymarin. During verification, the silymarin was replaced with silybin, and the method described in Example 1 of the patent was strictly followed.

(1) Preparation method (same as Example 1 of the original patent)

Dissolve 3 g of hydroxypropyl-β-cyclodextrin in 10 ml of water for injection, add 300 mg of silybin, adjust the pH to 10 with 1N NaOH solution, stir to dissolve the silybin, continue stirring for 5 hours, adjust the pH to 6.5 with 1N HCl solution, filter sterilize, divide into packages, freeze-dry, and seal to obtain the silybin freeze-dried powder injection of the comparative patent.

(2) Experimental phenomena and results

①Discoloration

During the process of adjusting the pH to 10 with NaOH solution and stirring to dissolve silibinin, the color of the solution gradually changed from an off-white suspension to a yellow-green and finally to a dark yellow clear solution. However, the preparation method of the present invention did not cause any color change during the entire preparation process.

② Destructive verification of silybin by NaOH

Adjusting the pH to 10 with NaOH solution to dissolve silibinin resulted in a noticeable color change in the resulting solution. This change in color may be due to the strong base, NaOH, damaging the silibinin structure. Therefore, we performed UV scanning on the alkaline solution to examine the appearance of new absorption peaks, thereby determining whether NaOH had damaged silibinin.

Raw material control solution:

Take an appropriate amount of silybin raw material, dissolve and dilute it with methanol to obtain a raw material control solution;

Compare with patented test solution:

Take an appropriate amount of the drug solution adjusted with NaOH solution to dissolve silybin, and dilute it with water to obtain the comparative patent test solution.

The test solution of the present invention:

Take the drug solution under Example 4 of the present invention and dilute it with water to obtain the test solution of the present invention.

The above solutions were scanned in the wavelength range of 190-600nm. The results showed that both the raw material control solution and the test solution of the present invention had maximum absorption at 287.5nm, and almost no absorption at other wavelengths (see Figures 1-4); while the comparative patent test solution had a larger absorption at 287.5nm and an even larger absorption peak at 326.0nm (see Figures 5 and 6). This shows that under the action of NaOH, the structure of silybin has undergone significant changes, while the drug solution prepared by the present invention has stable quality. Therefore, there are certain problems with the quality of the silymarin cyclodextrin inclusion compound prepared in this patent application, which may affect the safety and effectiveness of silybin.

③Cannot be completely reconstituted

The freeze-dried samples were reconstituted with water for injection, 0.9% sodium chloride injection, and 5% glucose injection, respectively. The results showed that all reconstituted solutions were significantly turbid, with the turbidity increasing with standing and precipitation occurring. The silybin injection of the present invention, however, exhibited excellent resolubility after freeze-drying, demonstrating a significant difference.

In summary, Chinese patent application CN02125823.6, entitled "Silymarin injection containing cyclodextrin or its derivatives," and publication number CN1391894A, describes an implementation scheme with poor feasibility. The cyclodextrin used is not the optimal match for silybin (see Example 1 of the present invention), and therefore a safe and stable silybin injection cannot be obtained.

Claims (5)

1. A lyophilized salymarin powder for injection, characterized in that it is prepared from the following components in a weight-volume ratio: The organic solvent for injection is polyethylene glycol 400. The cosolvent is distearylphosphatidylethanolamine-polyethylene glycol 2000. 2. The lyophilized salymarin powder for injection according to claim 1, characterized in that it is prepared from the following components in a weight-volume ratio: 3. The lyophilized samarin powder for injection according to any one of claims 1-2, wherein the lyophilization support is selected from one or more of mannitol, lactose, dextran 20, dextran 40, dextran 70, sucrose, xylitol, sorbitol, and trehalose. 4. The lyophilized samarin powder for injection according to any one of claims 1-2, wherein the pH adjuster is selected from one or more of citric acid, hydrochloric acid, glacial acetic acid, phosphoric acid, disodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium citrate, trisodium citrate, and sodium acetate. 5. The lyophilized silymarin powder for injection according to claim 1, characterized in that the preparation method comprises the following steps: weighing the prescribed amount of silymarin and an organic solvent for injection, adding a co-solvent, mixing, heating and stirring at 40-70°C to dissolve, obtaining an organic phase; weighing the prescribed amount of sulfobutyl ether-β-cyclodextrin and a lyophilization support agent, placing them in an appropriate amount of water for injection, stirring and dissolving, obtaining an aqueous phase; mixing the organic phase and the aqueous phase, mixing evenly, and then adjusting the volume to the total volume with water for injection to obtain a drug solution; adjusting the pH value of the drug solution to 2-7 with a pH adjuster; adding 0.05%-0.5% (by weight/volume) of activated carbon for injection, adsorbing for 15-60 minutes, and then filtering the solution through a 1μm titanium rod to remove carbon and a 0.22μm filter element for sterilization; dispensing, lyophilizing, and sealing to obtain the lyophilized silymarin powder for injection.