Preparation method and application of astragaloside lipidosome

Abstract

The invention belongs to the technical field of biological materials and biological medicines, and discloses a preparation method and application of astragaloside lipidosome. The invention discloses a preparation method of astragaloside lipidosome, which comprises the following steps: mixing phospholipids, a lipophilic additive, an oil-soluble solvent and an astragaloside active ingredient, and carrying out reduced pressure distillation to remove the oil-soluble solvent to obtain an astragaloside lipidosome film; and mixing the astragaloside lipidosome film with a water-based solvent, and carrying out hydration reaction to obtain the astragaloside lipidosome. The conditions of reduced pressure distillation are-1.5 to-0.5 MPa and 40-50 DEG C. The astragaloside lipidosome is prepared by adopting a simple film dispersion method, the stable astragaloside lipidosome is obtained by coating astragaloside with cheap and easily available raw materials such as phospholipids, a lipophilic additive and an oil-soluble solvent, the preparation cost is relatively low, the method is simple, the reaction conditions are mild, and the method is suitable for industrial production. Complicated equipment and process are not needed, and the implementation is easy.

Description

Technical Field

[0001] This invention belongs to the field of biomaterials and biomedicine, specifically relating to a method for preparing astragaloside A liposomes and its application. Background Technology

[0002] Astragaloside A is a natural saponin compound isolated and extracted from Astragalus membranaceus, a plant belonging to the genus Astragalus in the legume family. It is also the main component responsible for the medicinal effects of Astragalus membranaceus. Astragaloside A can regulate signal transduction pathways to exert various pharmacological effects, including anti-inflammatory, anti-fibrotic, anti-oxidative stress, anti-asthmatic, anti-diabetic, anti-tumor, immunomodulatory, and cardioprotective effects. However, astragaloside A has poor solubility in water and low bioavailability, which severely limits its application in the pharmaceutical field. To improve its bioavailability, it is necessary to select suitable carriers to expand its application range. Liposomes have a closed bilayer structure, can load both hydrophilic and lipophilic drugs, and possess nanoscale properties and good biocompatibility. As a drug carrier, they can improve drug stability, improve drug solubility, enhance drug targeting, and promote drug absorption. Preparing astragaloside A into liposomes is expected to overcome its existing defects and improve its medicinal value. Currently, some methods for preparing astragaloside A liposomes have been reported, but these methods suffer from problems such as complex preparation processes, high costs, and poor liposome stability. There is an urgent need for a simple, low-cost, and stable method for preparing astragaloside A liposomes. Summary of the Invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art, and to provide a new method for preparing astragaloside A liposomes that is simple and easy to operate. This method has the advantages of simple preparation process, low cost and good liposome stability.

[0004] The first objective of this invention is to provide a method for preparing astragaloside A liposomes.

[0005] The second aspect of the present invention aims to provide the application of the preparation method of the first aspect of the present invention in the preparation of products that improve drug bioavailability or improve drug sustained-release effect.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The first aspect of the present invention provides a method for preparing astragaloside A liposomes, comprising the following steps: mixing phospholipids, lipophilic additives, oil-soluble solvents and astragaloside A active ingredient, removing the oil-soluble solvent by vacuum distillation to obtain an astragaloside A liposome film; mixing the astragaloside A liposome film with an aqueous solvent and performing a hydration reaction to obtain astragaloside A liposomes; The conditions for vacuum distillation are -1.5 to -0.5 MPa and 40 to 50 °C. In some embodiments of the present invention, the phospholipids are selected from at least one of egg yolk lecithin, soybean lecithin, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, dioleoyl phosphatidylcholine, dipalmitoyl phosphatidic acid, dipalmitoyl phosphatidic acid, dipalmitoyl phosphatidylcholine, and dipalmitoyl phosphatidylglycerol; preferably egg yolk lecithin or soybean lecithin.

[0007] In some embodiments of the present invention, the lipophilic additive is selected from at least one of tocopherol, stearic acid, phosphatidic acid, ascorbic acid, cholesterol, cholesterol hemisuccinate and lanolin extract; preferably cholesterol.

[0008] In some embodiments of the present invention, the oil-soluble solvent is selected from at least one of methanol, ethanol, chloroform, dichloromethane, diethyl ether, propylene glycol, glycerol, isopropanol, dimethyl sulfoxide, and dimethylacetamide; preferably methanol, ethanol, or methanol-chloroform (mixed in equal proportions).

[0009] This invention obtains astragaloside A liposomes with high encapsulation efficiency and good stability by screening phospholipids and lipophilic additives.

[0010] In some embodiments of the present invention, the aqueous solvent is selected from water and / or phosphate buffer solution.

[0011] In some embodiments of the present invention, the active ingredient of astragaloside A includes astragaloside A, salts of astragaloside A, or lipids of astragaloside A.

[0012] In some embodiments of the present invention, the salts of astragaloside A include monovalent metal salts (such as sodium salts, potassium salts, etc.), divalent metal salts (such as magnesium salts, zinc salts, etc.), trivalent metal salts (such as aluminum salts), or mineral salts (such as phosphates, sulfates, etc.).

[0013] In some embodiments of the present invention, the esters of astragaloside A include fatty acid esters such as astragaloside methyl succinate, mineral esters such as phosphate esters, and aromatic esters such as benzoic acid.

[0014] In some embodiments of the present invention, the mass ratio of the phospholipids, lipophilic additives and astragaloside A active ingredient is (0.5~15):(0.1~10):1.

[0015] In some embodiments of the present invention, the mass ratio of the phospholipids, lipophilic additives and astragaloside A active ingredient is (5~15):(1~10):1.

[0016] In some embodiments of the present invention, the mass ratio of the phospholipids, lipophilic additives and astragaloside A active ingredient is (10~15):(1.5~5):1.

[0017] In some embodiments of the present invention, the mass-to-volume ratio (g / mL) of the astragaloside A active ingredient to the oil-soluble solvent is 1:(20~200).

[0018] In some embodiments of the present invention, the mass-to-volume ratio (g / mL) of the astragaloside A active ingredient to the oil-soluble solvent is 1:(50~200).

[0019] In some embodiments of the present invention, the mass-to-volume ratio (g / mL) of the astragaloside A active ingredient to the oil-soluble solvent is 1:(100~200).

[0020] In some embodiments of the present invention, the hydration reaction takes 1 to 6 hours.

[0021] In some embodiments of the present invention, the hydration reaction takes 1 to 5 hours.

[0022] In some embodiments of the present invention, the hydration reaction takes 2 to 4 hours.

[0023] In some embodiments of the present invention, the mass-volume ratio (g / mL) of the astragaloside A active ingredient to the aqueous solvent is 1:(20~200).

[0024] In some embodiments of the present invention, the mass-volume ratio (g / mL) of the astragaloside A active ingredient to the aqueous solvent is 1:(60~200).

[0025] In some embodiments of the present invention, the mass-volume ratio (g / mL) of the astragaloside A active ingredient to the aqueous solvent is 1:(100~200).

[0026] In some embodiments of the present invention, the conditions for vacuum distillation are -1 to -0.9 MPa and 40 to 45 °C.

[0027] This invention obtains astragaloside A liposomes with controllable particle size and a smooth, non-adhesive appearance by controlling the concentrations of various raw materials (phospholipids, lipophilic additives, oil-soluble solvents, etc.) and the active ingredient astragaloside A, as well as process parameters such as reaction conditions. This preparation method can improve the bioavailability of poorly water-soluble drugs and achieve targeted sustained release of drugs, thus expanding the application range of drugs.

[0028] A second aspect of the present invention provides the application of the preparation method of the first aspect of the present invention in the preparation of products that improve drug bioavailability or improve drug sustained-release effect.

[0029] In some embodiments of the present invention, the drug includes drugs with poor water solubility.

[0030] The beneficial effects of this invention are: This invention employs a simple thin-film dispersion method to prepare astragaloside A liposomes. It utilizes readily available and inexpensive raw materials such as phospholipids, lipophilic additives, and oil-soluble solvents to encapsulate astragaloside A, resulting in stable liposomes. The preparation cost is low, the method is simple, the reaction conditions are mild, and it requires no complex equipment or processes, making it easy to implement and conducive to large-scale production, thus generating significant economic benefits. Furthermore, the preparation method provided by this invention can produce astragaloside A liposomes with uniform and controllable particle size and excellent stability. These liposomes exhibit good water solubility, high stability, and a significant sustained-release effect, significantly improving the solubility of astragaloside A. In addition, the preparation method provided by this invention has universality; adding different drugs can yield liposomes encapsulated with different drugs, expanding its applicability. Attached Figure Description

[0031] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 The image shows an electron micrograph of the astragaloside A liposomes prepared in Example 4 of this invention, with a scale bar of 500 nm.

[0032] Figure 2 This is a particle size distribution diagram of the astragaloside A liposomes prepared in Example 4 of the present invention. Detailed Implementation

[0033] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0034] Unless otherwise specified in the examples, the procedures should be performed under standard conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0035] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0036] Example 1 A method for preparing astragaloside A liposomes includes the following steps: Weighing 0.1 g of soybean lecithin, 0.05 g of stearic acid, and 0.01 g of astragaloside A, adding them to 10 mL of methanol and stirring until fully mixed and dissolved to obtain a homogeneous mixture, removing the methanol from the mixture using a rotary evaporator to obtain an astragaloside A liposome film. Then, adding 20 mL of pure water and hydrating for 4 hours to obtain an astragaloside A liposome solution, thus obtaining the astragaloside A liposomes.

[0037] Example 2 A method for preparing astragaloside A liposomes includes the following steps: Weighing 0.1 g of soybean lecithin, 0.05 g of cholesterol, and 0.01 g of astragaloside A, adding them to 20 mL of anhydrous ethanol and stirring until fully dissolved to obtain a homogeneous mixture, removing methanol from the mixture using a rotary evaporator (-0.9 MPa, 45℃) to obtain an astragaloside A liposome film. Then, adding 20 mL of phosphate buffered saline (PBS) at pH 7.4 for hydration for 4 h to obtain an astragaloside A liposome solution, thus obtaining the astragaloside A liposomes.

[0038] Example 3 A method for preparing astragaloside A liposomes includes the following steps: Weighing 0.1 g of egg yolk lecithin, 0.05 g of cholesterol, and 0.01 g of astragaloside A, adding them to 20 mL of a mixed solution of methanol and chloroform (volume ratio 1:1), stirring to mix thoroughly and dissolve to obtain a homogeneous mixture, removing the methanol from the mixture using a rotary evaporator (-0.9 MPa, 45℃) to obtain an astragaloside A liposome film. Then, adding 20 mL of phosphate buffered saline (PBS) at pH 7.4 for hydration for 2 h to obtain an astragaloside A liposome solution.

[0039] Example 4 A method for preparing astragaloside A liposomes includes the following steps: Weighing 0.3 g of soybean lecithin, 0.03 g of cholesterol, and 0.02 g of astragaloside A, adding them to 20 mL of anhydrous ethanol and stirring until fully dissolved to obtain a homogeneous mixture, removing the ethanol from the mixture using a rotary evaporator (-0.9 MPa, 45℃) to obtain an astragaloside A liposome film. Then, adding 20 mL of pure water and hydrating for 4 h yields an astragaloside A liposome solution, thus obtaining the astragaloside A liposomes.

[0040] Effect Example 1. Structural characterization of astragaloside A liposomes The morphology, particle size, and potential value of the astragaloside A liposomes prepared in Examples 1-4 were characterized by transmission electron microscopy, zeta potential, and laser particle size analyzer.

[0041] The results showed that the astragaloside A liposomes prepared in Example 1 had a smooth elliptical bilayer structure with a particle size of 200 nm and a potential value of -6.0 mV; the astragaloside A liposomes prepared in Example 2 had a smooth circular bilayer structure with a particle size of 150 nm and a potential value of -8.6 mV; the astragaloside A liposomes prepared in Example 3 had a smooth circular bilayer structure with a particle size of 150 nm and a potential value of -8.6 mV; and the astragaloside A liposomes prepared in Example 4 had a smooth circular bilayer structure with a particle size of 200 nm and a potential value of -8.6 mV. Figure 1 and Figure 2 ).

[0042] 2. Determination of encapsulation efficiency of astragaloside A liposomes The encapsulation efficiency of astragaloside A in the liposomes prepared in Examples 1-4 was determined by high performance liquid chromatography.

[0043] The results showed that the encapsulation efficiency of the astragaloside A liposomes prepared in Example 1 was 65%; the encapsulation efficiency of the astragaloside A liposomes prepared in Example 2 was 78%; the encapsulation efficiency of the astragaloside A liposomes prepared in Example 3 was 70%; and the encapsulation efficiency of the astragaloside A liposomes prepared in Example 4 was 87%.

[0044] 3. Assay for sustained-release of astragaloside A liposomes The cumulative release of astragaloside A from the liposomes prepared in Examples 1-4 was determined by HPLC within 72 h, and the release rate of astragaloside A was calculated according to the following formula.

[0045] Release rate (%) = [ ] / W 总 × 100%.

[0046] Among them, C n V represents the concentration of astragaloside A in the dialysis medium at a specified sampling time point, V represents the total volume of the dialysis medium, Wtotal is the initial total amount of astragaloside A in the test sample, and Ctotal represents the total concentration of astragaloside A in the dialysis medium at a specified sampling time point. i V represents the concentration of astragaloside A in the dialysis medium at the previous sampling point. i This represents the sampling volume.

[0047] The results showed that the drug release rate of the astragaloside A liposomes prepared in Example 1 was 30% at 72 h; the drug release rate of the astragaloside A liposomes prepared in Example 2 was 50% at 72 h; the drug release rate of the astragaloside A liposomes prepared in Example 3 was 41% at 72 h; and the drug release rate of the astragaloside A liposomes prepared in Example 4 was 58% at 72 h (Table 1).

[0048] Table 1. Release of astragaloside A and drug-loaded liposomes in Example 4

[0049] 4. Determination of storage stability of astragaloside A liposomes The astragaloside A liposomes prepared in Examples 1-4 were stored in a refrigerator at 4°C, and the presence of abnormal phenomena such as aggregation and turbidity during storage was observed.

[0050] The results showed that the astragaloside A liposomes prepared in Example 1 showed no abnormalities after being stored at 4°C for 7 days; the astragaloside A liposomes prepared in Example 2 showed no abnormalities after being stored at 4°C for 20 days; the astragaloside A liposomes prepared in Example 3 showed no abnormalities after being stored at 4°C for 10 days; and the astragaloside A liposomes prepared in Example 4 showed no abnormalities after being stored at 4°C for 10 days.

[0051] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention. Furthermore, unless otherwise specified, the embodiments of the present invention and the features thereof can be combined with each other.

Claims

1. A method for preparing astragaloside A liposomes, comprising the following steps: Phospholipids, lipophilic additives, oil-soluble solvents, and astragaloside active ingredients are mixed, and the oil-soluble solvent is removed by vacuum distillation to obtain astragaloside liposome film; the astragaloside liposome film is mixed with an aqueous solvent and hydrated to obtain astragaloside liposomes. The conditions for vacuum distillation are -1.5 to -0.5 MPa and 40 to 50 °C.

2. The preparation method according to claim 1, characterized in that, The phospholipids are selected from at least one of egg yolk lecithin, soybean lecithin, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, dioleoyl phosphatidylcholine, dipalmitoyl phosphatidic acid, dipalmitoyl phosphatidic acid, dipalmitoyl phosphatidylcholine, and dipalmitoyl phosphatidylglycerol.

3. The preparation method according to claim 1, characterized in that, The lipophilic additive is selected from at least one of tocopherol, stearic acid, phosphatidic acid, ascorbic acid, cholesterol, cholesterol hemisuccinate, and lanolin extract.

4. The preparation method according to any one of claims 1 to 3, characterized in that, The oil-soluble solvent is selected from at least one of methanol, ethanol, chloroform, dichloromethane, diethyl ether, propylene glycol, glycerol, isopropanol, dimethyl sulfoxide, and dimethylacetamide.

5. The preparation method according to claim 4, characterized in that, The aqueous solvent is selected from water and / or phosphate buffer solution.

6. The preparation method according to claim 4, characterized in that, The active ingredient of astragaloside A includes astragaloside A, its salts, or its esters.

7. The preparation method according to any one of claims 1 to 3, characterized in that, The mass ratio of the phospholipids, lipophilic additives and astragaloside A active ingredient is (0.5~15):(0.1~10):

1.

8. The preparation method according to claim 7, characterized in that, The mass-to-volume ratio (g / mL) of the active ingredient astragaloside A to the oil-soluble solvent is 1:(20~200).

9. The preparation method according to claim 7, characterized in that, The hydration reaction takes 1 to 6 hours.

10. The application of the preparation method according to any one of claims 1 to 9 in the preparation of products that improve drug bioavailability or improve drug sustained-release effect.

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