CN114224841B - WGX-50 liposome, preparation method and application - Google Patents

WGX-50 liposome, preparation method and application Download PDF

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CN114224841B
CN114224841B CN202210074080.XA CN202210074080A CN114224841B CN 114224841 B CN114224841 B CN 114224841B CN 202210074080 A CN202210074080 A CN 202210074080A CN 114224841 B CN114224841 B CN 114224841B
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wgx
liposome
carrying
30min
phospholipids
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CN114224841A (en
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琚建伟
张守兵
丁保安
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Shanghai Tongyan Industrial Co ltd
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Abstract

The invention discloses a WGX-50 liposome, a preparation method and application thereof, wherein the WGX-50 liposome comprises the following components: WGX-50, phospholipids, phytosterols, lipid structure modifiers, stabilizing additives and deionized water; the preparation method comprises the following steps: dispersing WGX-50, phospholipids and phytosterols in an organic solvent; rotary evaporation and vacuum drying, removing the organic solvent and forming a dried film; adding a stabilizing additive and deionized water into the dried film to carry out hydration treatment and micronization treatment; adding the aqueous solution of the lipid structure modifier into the solution obtained by the micronization treatment, mixing and stirring to obtain the WGX-50 liposome. According to the invention, the WGX-50 is wrapped in a liposome form, so that the water solubility of the WGX-50 can be effectively improved, and the application potential of the WGX-50 is improved; meanwhile, the stability of the prepared WGX-50 lipid is obviously improved through the combined use of phytosterols, lipid structure modifiers and stability additives.

Description

WGX-50 liposome, preparation method and application
Technical Field
The invention relates to the technical fields of cosmetics, health-care products and medicines, in particular to a WGX-50 liposome, a preparation method and application thereof.
Background
Zanthoxylum bungeanum Maxim WGX-50 is a natural ingredient existing in natural seasoning Zanthoxylum bungeanum Maxim, has multiple cosmetic functions of potential moisture preservation, oxidation resistance and the like, and can be used for developing various cosmetics. The research on WGX-50 also has a specific gene regulation effect, can effectively inhibit the aggregation of Abeta amyloid in human brain (the aggregation of Abeta amyloid in brain is considered as an important pathological feature of Alzheimer's disease) and the expression of lipid apoptosis protein, and reduce apoptosis, thereby effectively protecting nerve cells, and has potential application in preparing anti-Alzheimer's disease drugs. In 2010 Wei Dongqing et al, 3, 4-dimethoxy benzaldehyde, nitromethane and cinnamic acid are taken as raw materials, and the three-step reaction of condensation, reduction and condensation amidation is adopted to realize the artificial synthesis of the WGX-50, so that the defects of low content of extracted target molecules in pepper, high extraction cost and the like are overcome, and a solid foundation is laid for the application of the WGX-50 in a plurality of aspects; however, WGX-50 has disadvantages of poor water solubility and low solubility in application, and further improvement is needed.
The liposome is a double-molecular-layer closed vesicle which is formed by taking phospholipid as a main material, and is discovered and proposed by British scientist Bangham in 1961, and the structural characteristics of the double-molecular-layer closed vesicle can be used for loading fat-soluble and water-soluble medicines, and meanwhile, the liposome has the advantages of biocompatibility, biodegradability, non-toxicity, non-immunogenicity and the like due to the structure similar to a biological membrane. Since the beginning of the 70 th century, liposomes became a drug carrier, and because of their rapid development as new dosage forms of drugs, nearly hundreds of liposome formulations have been marketed. With the intensive research, the application potential of the liposome in cosmetics is gradually developed, and besides the transdermal rate of active substances is increased, the liposome cosmetics can also effectively protect active ingredients unstable in light, heat and oxidation, and improve the activity and stability.
Although the prior art has conducted intensive studies on liposome technology of poorly soluble substances, the research on encapsulation of WGX-50 is relatively small; in addition, the WGX-50 liposome prepared by the prior method has the characteristics of poor water solubility and poor fat solubility, and the prepared liposome is easy to have the problems of aggregation, coarseness, crystallization, delamination and the like.
Disclosure of Invention
In order to solve the problems of poor water solubility and poor applicability of WGX-50 in the prior art, the liposome prepared by adopting a conventional method also has the problems of easy aggregation, easy crystallization and the like, the invention provides the WGX-50 liposome, a preparation method and application thereof.
A first object of the present invention is to provide a WGX-50 liposome comprising: WGX-50, phospholipids, phytosterols, lipid structure modifiers, stabilizing additives and deionized water.
As a further improvement of the present invention, the WGX-50 has the structural formula:
Figure BDA0003483300510000021
as a further improvement of the present invention, the phospholipids include one or more of hydrogenated lecithin, soybean lecithin, hydroxylated lecithin, lysolecithin, hydrogenated phosphatidylcholine, phosphatidylcholine and hydrogenated lysolecithin.
As a further improvement of the present invention, the phospholipids are hydrogenated lecithin and/or soybean lecithin.
As a further improvement of the invention, the phytosterols include one or more of phytosterol oleate, phytosterol/octyldodecanol lauroyl glutamate and phytosterol macadamia oleate.
As a further improvement of the present invention, the lipid structure modifier includes one or more of chitin, chitosan, carboxymethyl chitosan, chitosan.
As a further improvement of the present invention, the stabilizing additive comprises one or more of ethoxydiglycol, bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate, polyglycerol-10 eicosadioate/tetradecanedioate.
As a further development of the invention, the stabilizing additive is ethoxydiglycol and/or bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate.
As a further improvement of the present invention, the phospholipids: phytosterols: WGX-50: stabilizing additive: the mass percentage of the lipid structure modifier is 8-20: 1 to 6:1 to 10:2 to 8:5 to 15.
A second object of the present invention is to provide a method for preparing WGX-50 liposome, comprising:
s1, dispersing WGX-50, phospholipids and phytosterols in an organic solvent according to a preset proportion; wherein the organic solvent comprises one of ethanol, ethyl acetate and chloroform;
s2, removing the organic solvent by rotary evaporation at a preset temperature;
s3, vacuum drying at room temperature, and removing residual organic solvent to form a dried film;
s4, adding a stabilizing additive and deionized water into the dried film to carry out hydration treatment;
s5, carrying out micronization treatment on the hydrated product by using an ultrasonic cell disruptor;
s6, adding the aqueous solution of the lipid structure modifier into the solution obtained by the micronization treatment, mixing and stirring to obtain the WGX-50 liposome.
The third object of the present invention is to provide an application of the WGX-50 liposome in preparing cosmetics, health products and medicines.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the WGX-50 is wrapped in a liposome form, so that the water solubility of the WGX-50 can be effectively improved, and the application potential of the WGX-50 is improved; meanwhile, the stability of the prepared WGX-50 lipid can be obviously improved through the combined use of phytosterols, lipid structure modifiers and stability additives.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The present invention provides a WGX-50 liposome comprising: WGX-50, phospholipids, phytosterols, lipid structure modifiers, stabilizing additives and deionized water;
wherein, the liquid crystal display device comprises a liquid crystal display device,
the structural formula of the WGX-50 of the invention is as follows:
Figure BDA0003483300510000031
the phospholipids of the present invention include one or more of hydrogenated lecithin, soybean lecithin, hydroxylated lecithin, lysolecithin, hydrogenated phosphatidylcholine, phosphatidylcholine and hydrogenated lysolecithin, preferably the above-mentioned single phospholipids, and further preferably hydrogenated lecithin or soybean lecithin.
The plant sterols of the present invention include one or more of plant sterol oleate, plant sterol/octyldodecanol lauroyl glutamate and plant sterol macadamia nut oleate, preferably a single plant sterol as described above.
The lipid structure modifier of the present invention includes one or more of chitin, chitosan, carboxymethyl chitosan, and preferably the above-mentioned single lipid structure modifier.
The stabilizing additive of the present invention comprises one or more of ethoxydiglycol, bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate, polyglycerol-10 eicosadioate/tetradecanedioate, preferably the above-mentioned single stabilizing additive, further preferably ethoxydiglycol or bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate.
Phospholipids of the present invention: phytosterols: WGX-50: stabilizing additive: the mass percentage of the lipid structure modifier is 8-20: 1 to 6:1 to 10:2 to 8:5 to 15.
The invention provides a preparation method of WGX-50 liposome, comprising the following steps:
s1, dissolving:
dispersing WGX-50, phospholipids and phytosterols in an organic solvent according to the preset proportion, and carrying out ultrasonic dissolution;
wherein the organic solvent is one of ethanol, ethyl acetate and chloroform, and the ultrasonic dissolution time is 1-4 min, preferably 2min;
s2, evaporating:
at a preset temperature, slowly rotary evaporating to remove the organic solvent;
wherein the evaporation temperature is 30-50 ℃, preferably 40 ℃; the revolution of the rotary evaporator is 60-100 rpm, preferably 80rpm;
s3, vacuum drying:
vacuum drying at room temperature to further remove residual small amount of organic solvent and form a dried film;
wherein the vacuum drying time is 20-40 min, preferably 30min;
s4, hydration treatment:
adding a stabilizing additive and deionized water into the dry film according to a preset proportion, and carrying out rotary hydration to obtain liposome suspension;
wherein the treatment time of the rotary hydration is 20-40 min, preferably 30min;
s5, micronizing:
transferring the liposome suspension to a cell pulverizer for ultrasonic micronization treatment;
wherein the ultrasonic treatment time is 5-15 min, preferably 10min;
s6, lipid structure modification:
adding the water solution of the lipid structure modifier into the clarified solution obtained by the micronization treatment, and mixing and stirring to obtain WGX-50 liposome;
wherein, the mixing and stirring time is 20-40 min, preferably 30min.
The WGX-50 liposome prepared by the invention can be used for preparing cosmetics, health care products and medicines.
Example 1
Accurately weighing 80mg of soybean lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 20mg of bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylic acid ester, carrying out rotary hydration for 30min to obtain liposome suspension, carrying out ultrasonic treatment under a cell pulverizer for 10min, adding 10mL of aqueous solution containing 50mg of chitin into the obtained clear solution, mixing and stirring for 30min, and obtaining uniform and transparent WGX-50 liposome solution.
Example 2
Accurately weighing 80mg of soybean lecithin, 10mg of phytosterol/octyl dodecanol lauroyl glutamate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 20mg of bis-diethoxy diglycol cyclohexane 1, 4-dicarboxylic acid ester, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of chitin into the obtained clear solution, mixing and stirring for 30min, and obtaining uniform and transparent WGX-50 liposome solution.
Example 3
Accurately weighing 80mg of soybean lecithin, 10mg of phytosterol macadamia nut oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a rotary evaporator at 40 ℃, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 20mg of bis-diethoxy diglycol cyclohexane 1, 4-dicarboxylate, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of chitin into the obtained clear solution, mixing and stirring for 30min to obtain uniform and transparent WGX-50 liposome solution.
Example 4
Accurately weighing 80mg of hydrogenated lecithin, 4mg of phytosterol oleate and 4mg of WGX-50 in a 50mL flask, adding 20mL of chloroform, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the chloroform, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 20mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to the cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Example 5
Accurately weighing 80mg of soybean lecithin, 20mg of phytosterol oleate and 6mg of WGX-50 in a 50mL flask, adding 20mL of ethyl acetate, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the ethyl acetate, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 20mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining the uniform WGX-50 liposome solution.
Example 6
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining uniform and transparent WGX-50 liposome solution.
Example 7
Accurately weighing 100mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of polyglycerol-10 eicosadioate/tetradecanedioate, carrying out rotary hydration for 30min to obtain liposome suspension, carrying out ultrasonic treatment under a cell pulverizer for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained solution, mixing and stirring for 30min to obtain uniform WGX-50 liposome solution.
Example 8
Accurately weighing 100mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of polyglycerol-10 eicosadioate/tetradecanedioate, carrying out rotary hydration for 30min to obtain liposome suspension, carrying out ultrasonic treatment under a cell pulverizer for 10min, adding 10mL of aqueous solution containing 50mg of chitosamine into the obtained solution, mixing and stirring for 30min, and obtaining the uniform WGX-50 liposome solution.
Example 9
Accurately weighing 120mg of hydrogenated lecithin, 40mg of phytosterol oleate and 10mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of polyglycerol-10 eicosadioate/tetradecanedioate, carrying out rotary hydration for 30min to obtain liposome suspension, carrying out ultrasonic treatment under a cell pulverizer for 10min, adding 10mL of aqueous solution containing 50mg of chitosan into the obtained solution, mixing and stirring for 30min, and obtaining the uniform WGX-50 liposome solution.
Comparative example 1
Accurately weighing 80mg of soybean lecithin and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a rotary evaporator at 40 ℃, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the flask, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, moving to the position under a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained solution, mixing and stirring for 30min, and obtaining transparent WGX-50 liposome solution.
Comparative example 2
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water, carrying out rotary hydration for 30min to obtain liposome suspension, moving to the position under a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained solution, mixing and stirring for 30min, and obtaining the WGX-50 liposome solution.
Comparative example 3
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 20mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, and carrying out ultrasonic treatment under a cell pulverizer for 10min to obtain the WGX-50 liposome solution.
Comparative example 4
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the bottle wall, carrying out vacuum drying at room temperature for 30min, adding 20mL of deionized water, carrying out rotary hydration for 30min to obtain liposome suspension, and carrying out ultrasonic treatment under a cell pulverizer for 10min to obtain the WGX-50 liposome solution.
Comparative example 5
Accurately weighing 80mg of hydrogenated lecithin, 10mg of cholesterol and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to the cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 6
Accurately weighing 80mg of hydrogenated lecithin, 2mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 7
Accurately weighing 80mg of hydrogenated lecithin, 80mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 8
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of polyethylene glycol 400, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to the cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 9
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of Tween 80, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to the cell pulverizer, carrying out ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 10
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 50mg of sodium alginate into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 11
Accurately weighing 80mg of hydrogenated lecithin, 10mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 20mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
Comparative example 12
Accurately weighing 80mg of hydrogenated lecithin, 2mg of phytosterol oleate and 8mg of WGX-50 in a 50mL flask, adding 20mL of absolute ethyl alcohol, carrying out ultrasonic dissolution for 2min, placing under a 40 ℃ rotary evaporator, slowly evaporating at 80rpm to remove the absolute ethyl alcohol, forming a layer of uniform film on the wall of the bottle, carrying out vacuum drying at room temperature for 30min, adding 10mL of deionized water and 40mg of ethoxydiglycol, carrying out rotary hydration for 30min to obtain liposome suspension, transferring to a cell pulverizer for ultrasonic treatment for 10min, adding 10mL of aqueous solution containing 200mg of carboxymethyl chitosan into the obtained clear solution, mixing and stirring for 30min, and obtaining transparent uniform WGX-50 liposome solution.
And (3) verification:
the above 21 parts of liposome (examples 1 to 9 and comparative examples 1 to 12) solutions were each uniformly divided into 3 parts, and placed in a refrigerator at 4℃and an incubator at room temperature and 45℃for 48 hours, and the state of the solutions before and after the placement was observed, and the stability thereof was compared, and the results are shown in Table 1;
TABLE 1
Figure BDA0003483300510000101
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Figure BDA0003483300510000111
As can be seen from Table 1, in the preparation process of the liposome, the phytosterols, the stabilizing additive and the lipid structure modifier are all indispensable, and different use amounts and combination matching have great influence on the transparency and stability of the liposome solution, and the reasonable matching proportion can effectively increase the stability of the WGX-50 liposome.
The factors influencing stability are analyzed as follows:
the polar group in the phytosterol molecule can interact with the hydrophilic polar head in the phospholipid molecular structure, so that the phase transition temperature of the phospholipid and the charge on the surface of the phospholipid bilayer are improved, and the stability of the liposome vesicle is improved; the consumption of the phytosterol is too low, the acting force with phospholipid molecules is insufficient, the phase transition temperature of the phospholipid cannot be effectively improved, and the surface charge of a bilayer is limited; and too high an amount will affect the formation of vesicles of phospholipid bilayer, so that the reasonable ratio of phytosterols to phospholipids is also a key factor affecting the stability of the liposome.
The lipid structure modifier is a polymer, can be wrapped on the surface of the whole phospholipid molecular vesicle through the actions of electrostatic attraction and the like after being dissolved and dissociated in water, increases the surface charge of the phospholipid molecular vesicle, increases the repulsive force among the vesicles, effectively slows down the coalescence among the vesicles, and increases the stability of the liposome; the amount of the lipid structure modifier to be used should be within a proper range for addition, while desired stability is obtained. The use level is low, polymers such as chitin and the like can not wrap all liposome vesicles well and effectively, and the use level is excessive, and long-chain chitin fragments on the surface of the liposome can interact with each other to generate bridging flocculation, so that the stability of the liposome is also influenced.
The stability additives ethoxydiglycol, bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate, polyglycerol-10 eicosadioate/tetradecanedioate can increase the solubility of WGX-50 in vesicles by dipole-dipole interaction, and further improve the stability of the liposomes by blocking coalescence among vesicles due to its adsorption to the surface of the liposome vesicles or steric hindrance by its hydrophobic chain.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (7)

1. A WGX-50 liposome, comprising: WGX-50, phospholipids, phytosterols, lipid structure modifiers, stabilizing additives and deionized water; wherein, the liquid crystal display device comprises a liquid crystal display device,
the lipid structure modifier is one or more of chitin, chitosan, carboxymethyl chitosan and chitosan;
the stabilizing additive is one or more of ethoxydiglycol, bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylic acid ester and polyglycerol-10 eicosadioate/tetradecanedioate;
the phospholipids: phytosterols: WGX-50: stabilizing additive: the mass percentage of the lipid structure modifier is 8-20: 1 to 6:1 to 10:2 to 8:5 to 15.
2. The WGX-50 liposome of claim 1, wherein the phospholipids comprise one or more of hydrogenated lecithin, soy lecithin, hydroxylated lecithin, lysolecithin, hydrogenated phosphatidylcholine, and hydrogenated lysolecithin.
3. The WGX-50 liposome of claim 2, wherein the phospholipids are hydrogenated lecithin and/or soy lecithin.
4. The WGX-50 liposome of claim 1, wherein the phytosterols comprise one or more of phytosterol oleate, phytosterol/octyldodecanol lauroyl glutamate, and phytosterol macadamia oleate.
5. The WGX-50 liposome of claim 1, wherein the stabilizing additive is ethoxydiglycol and/or bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate.
6. A method of preparing WGX-50 liposome according to any one of claims 1 to 5, comprising:
s1, dispersing WGX-50, phospholipids and phytosterols in an organic solvent according to a preset proportion; wherein the organic solvent comprises one of ethanol, ethyl acetate and chloroform;
s2, removing the organic solvent by rotary evaporation at a preset temperature;
s3, vacuum drying at room temperature, and removing residual organic solvent to form a dried film;
s4, adding a stabilizing additive and deionized water into the dried film to carry out hydration treatment;
s5, carrying out micronization treatment on the hydrated product by using an ultrasonic cell disruptor;
s6, adding the aqueous solution of the lipid structure modifier into the solution obtained by the micronization treatment, mixing and stirring to obtain the WGX-50 liposome.
7. Use of a WGX-50 liposome according to any one of claims 1 to 5, in the manufacture of cosmetics, health products and medicaments.
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