CN107669638B - PEG-PCL-PEG triblock copolymer modified madecassoside liposome and application thereof - Google Patents

PEG-PCL-PEG triblock copolymer modified madecassoside liposome and application thereof Download PDF

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CN107669638B
CN107669638B CN201710995253.0A CN201710995253A CN107669638B CN 107669638 B CN107669638 B CN 107669638B CN 201710995253 A CN201710995253 A CN 201710995253A CN 107669638 B CN107669638 B CN 107669638B
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刘美凤
董礁礁
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South China University of Technology SCUT
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Abstract

The invention discloses a PEG-PCL-PEG triblock copolymer modified madecassoside liposome and application thereof, belonging to the fields of pharmaceutical preparations, skin care products and daily chemicals. The liposome comprises PEG-PCL-PEG triblock polymer, madecassoside, cholesterol and lipid components; the madecassoside is encapsulated in liposomes. The invention uses PEG-PCL-PEG triblock polymer to modify MA liposome, can improve the bonding effect of the drug and the skin, particularly can change the drug into paste when being applied to the skin from liquid, plays the role of a drug storage, prolongs the action time of the MA and the skin, has long-term slow release and enhances the treatment effect. The method has simple preparation process, is easy to amplify, and has wide application prospect for future large-scale medicine production and potential medical application thereof.

Description

PEG-PCL-PEG triblock copolymer modified madecassoside liposome and application thereof
Technical Field
The invention belongs to the fields of pharmaceutical preparations, skin care products and daily chemicals, and particularly relates to a PEG-PCL-PEG triblock copolymer modified madecassoside liposome and application thereof.
Background
Madecassoside (MA) is a triterpene saponin in centella asiatica extract, and has good water solubility, structure containing trisaccharide chain, and large molecular weight. The traditional Chinese medicine composition is used as a main component of a medicament widely applied to treating skin wounds such as surgical wounds, scalds and burns in clinic, and has various pharmacological effects of inhibiting bacteria, diminishing inflammation, promoting fibroblast regeneration and the like. However, because they are polar macromolecular substances, they are not easy to penetrate the epidermis of the skin to reach the affected part, so that the drug effect and application thereof are limited.
Liposomes (liposomes) are vesicles that spontaneously form in water from phospholipids and can encapsulate drugs. The medicine carrier is usually used as a medicine carrier because the preparation process is simple, and the medicine carrier has the characteristics of no toxicity, no immunogenicity, degradability, slow release, targeting and the like in a human body, and the medicine is delivered into the interior of cells or tissue gaps by utilizing the characteristic of high structural similarity of the medicine carrier and cell membranes, so that the medicine carrier can be used for the transgenic technology or the medicine preparation.
The skin acts as a first barrier to protect the body and also provides a barrier to the effective permeation of drugs. The liposome is selected as a carrier to wrap the medicine, and the medicine is targeted to the skin, so that the capability of the medicine penetrating through the skin is enhanced, and the effective healing of skin wounds is promoted.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention mainly aims to provide the madecassoside liposome modified by the PEG-PCL-PEG triblock copolymer.
In order to better exert the effect of promoting wound healing of MA liposome, the invention provides a technology for preparing modified liposome with better skin adhesiveness and scald treatment effect by an emulsion method aiming at the characteristic of easy flowing during skin administration.
The PECE modified liposome adopts an emulsion method to encapsulate MA in the liposome. The liposome has long-acting slow release effect, and is beneficial to playing the roles of bacteriostasis, inflammation diminishing and fibroblast regeneration promoting; can effectively penetrate epidermis, so that more medicinal components stay in dermis, bioavailability is improved, and wound healing is promoted. Moreover, the addition of PECE can enhance the adhesion between the liposome and the skin and better play the role of long-acting and slow-release.
The invention also aims to provide a preparation method of the PEG-PCL-PEG triblock copolymer modified madecassoside liposome.
The invention also aims to provide the application of the PEG-PCL-PEG triblock copolymer modified madecassoside liposome.
The purpose of the invention is realized by the following technical scheme:
a PEG-PCL-PEG triblock copolymer modified madecassoside liposome, which comprises PECE (PEG-PCL-PEG triblock copolymer), Madecassoside (MA), cholesterol and lipid component; the madecassoside is encapsulated in a liposome; the liposome prepared by the invention can be converted into gel state or paste state by liquid when being applied to skin, has good skin adhesion and scald treatment effect, and is very suitable for skin administration.
The lipid component is phospholipid including natural phospholipid or synthetic phospholipid, specifically including egg yolk lecithin, soybean lecithin, HSPC (hydrogenated soybean lecithin), etc.
The preparation method of the PEG-PCL-PEG triblock copolymer modified madecassoside liposome comprises the following steps:
(1) weighing PEG-PCL-PEG triblock polymer, lipid component, cholesterol and a small amount of antioxidant, and dissolving in a certain amount of organic solvent to obtain organic phase solution;
(2) adding the aqueous phase solution containing the madecassoside into the organic phase solution obtained in the step (1), and stirring to form a W/O emulsion, or adding the W/O emulsion into a second aqueous phase, and stirring to form a W/O/W multiple emulsion;
(3) evaporating the W/O emulsion or W/O/W multiple emulsion in the step (2) under reduced pressure to remove the organic solvent, then drying in vacuum to remove the residual organic solvent in the emulsion, and filtering the membrane to obtain the PEG-PCL-PEG triblock copolymer modified madecassoside liposome; storing at 4 deg.C for use.
Preferably, the lipid component in step (1) is a phospholipid, preferably egg yolk lecithin, soybean lecithin, HSPC (hydrogenated soybean lecithin), etc.; the mass ratio of the PEG-PCL-PEG triblock polymer to the lipid component is (0.5-1): 1, preferably (0.8-1): 1, more preferably 1: 1; the antioxidant is vitamin C, BHA (butyl hydroxy anisol) and vitamin E acetate (V)EAcetate esters); the organic solvent is preferably diethyl ether, dichloromethane, chloroform, etc.
The mass ratio of the lipid component to the cholesterol is 5: 1.
preferably, the concentration of cholesterol in the organic phase solution in the step (1) is 10 to 50mg/mL, preferably 10 mg/mL.
Preferably, the concentration of the antioxidant in the organic phase solution in the step (1) is 5 mL/L;
preferably, the water phase or the second water phase in the step (2) is an aqueous solution containing madecassoside or a PBS solution, and the concentration is 1-160 mg/mL, preferably 20 mg/mL; the stirring time is 1-30 min, preferably 1-10 min, and more preferably 5 min; the stirring speed is 100-1000 rpm, preferably 400-1000 rpm, more preferably 500 rpm; the second stirring time of the multiple emulsion method is 1-30 min, preferably 3-5 min, and more preferably 5 min; the stirring speed is 100-1000 rpm, preferably 500-800 rpm, and more preferably 500 rpm.
Preferably, the pore diameter of the filter membrane in the step (3) is 0.22-1 μm, preferably 0.45 μm; the madecassoside liposome prepared by W/O emulsion is preferably oily membrane filter, and the MA liposome prepared by W/O/W multiple emulsion is preferably aqueous membrane filter, more preferably polycarbonate membrane filter.
The preparation method of the PEG-PCL-PEG triblock copolymer in the step (1) comprises the following steps:
adding caprolactone (epsilon-CL) and polyethylene glycol monomethyl ether (MPEG), under the action of a catalyst, breaking ester bonds in the epsilon-CL, reacting with hydroxyl in the MPEG to generate new ester bonds, namely PEG-PCL fragments (first step reaction), then adding hexamethylene diisocyanate (HMDI), continuously reacting isocyanate groups in the HMDI with the hydroxyl generated in the PEG-PCL to generate amide bonds (second step reaction), purifying to obtain a PEG-PCL-PEG triblock Polymer (PECE), and drying in vacuum to constant weight for later use; the polymer is a biodegradable temperature sensitive material.
First step reaction raw material ratio n(MPEG)：n(ε-CL)Is 1: (1 to 15), preferably 1: (5-15), more preferably 1: 10; the reaction conditions are that the reaction is carried out for 4-15 h at 100-200 ℃, and preferably for 12h at 120 ℃;
the ratio of raw materials in the second step n(ε-CL)：n(HMDI)The ratio of (1-15): 1, preferably 10: 1; reaction conditionsReacting for 3-9 h at 50-100 ℃, preferably for 6h at 80 ℃;
under the optimized condition, the first step reaction product PEG-PCL and the second step reaction product PEG-PCL-PEG are respectively subjected to infrared spectrum characterization.
The catalyst is stannous octoate.
The PEG-PCL-PEG triblock copolymer modified madecassoside liposome is applied to the technical field of pharmaceutical preparations or skin care cosmetics.
Preferably, the application of the madecassoside liposome in preparing a transdermal or mucosal absorption pharmaceutical preparation for treating skin scald, skin cut, mucosal injury or scar;
preferably, the madecassoside liposome is applied to the preparation of skin care cosmetics for scar repair, deep skin damage or anti-aging, anti-oxidation or whitening and beautifying.
The mechanism of the invention is as follows:
the PEG-PCL-PEG triblock copolymer with lower molecular weight can generate the conversion between a solution state and a gel state after the aqueous solution in a specific concentration range is stimulated by external environment (such as temperature, PH, pressure and the like), and has wide application prospect in the in-situ gel field of drug controlled release, cell embedding, tissue repair and the like. The MA liposome modified by PEG-PCL-PEG shows good skin adhesiveness and scald treatment effect in a rat secondary scald test.
The liposome drug-loaded system has good biocompatibility with skin tissues, can be used for carrying drugs to permeate the epidermis and delivering the drugs to the part to be treated by the dermis, and can be used as an external preparation to be applied to the skin, however, the liposome has high fluidity, so that the application is limited. Therefore, aiming at the defects that a medicinal solution or a liposome preparation thereof has stronger fluidity, the medicament is easy to lose, and the medicament is difficult to stay on the surface of the skin for a long time, and the like, the PEG-PCL-PEG (PECE) triblock copolymer temperature-sensitive material is synthesized, the (PECE) is uniformly added in the process of preparing the Madecassoside (MA) liposome by adopting an emulsion method, the MA liposome modified by the PECE is obtained by optimizing the synthesis condition of the PECE and the proportion of the PECE and the lipid, has proper phase transition temperature, is in a gel state or a semisolid paste state at the body surface temperature, is prepared into the PEG-PCL-PEG modified madecassoside liposome with good skin adhesiveness and scald treatment effect, and the optimal preparation process is optimized.
Compared with the prior art, the invention has the following advantages and effects:
(1) the invention takes an emulsifier method as a basis, and skillfully applies the temperature-sensitive material PECE to the preparation process of the MA liposome, thereby improving the problems of easy flowing of the MA liposome, inconvenient medicine application and poor skin adhesion; the PECE modified MA liposome is in a gel state or a paste state, is convenient for skin administration, and has a treatment effect obviously higher than that of the MA liposome in a rat scald experiment.
(2) The phase transition temperature of the PECE modified MA liposome prepared by the invention is 18-32 ℃ when the liquid state is converted into the gel state or the paste state, and the characteristic of the invention obviously solves the problems that the MA solution or the conventional MA liposome is easy to lose when being applied to the skin due to strong fluidity.
(3) Compared with a common skin local application preparation, the PECE modified MA liposome transdermal drug delivery system has long-acting slow release property, and after the MA liposome penetrates through the horny layer, drug depots can be formed in the epidermis and the dermis, so that the drugs are slowly released, and the treatment effect on a wound lesion part can be directly and durably realized.
(4) The invention modifies the MA liposome by PECE, which can improve the bonding effect of the drug and the skin, especially the drug can be changed into paste by liquid when being applied to the skin, which plays the role of drug storage, prolongs the action time of MA and the skin, and enhances the treatment effect. The method has simple preparation process, is easy to amplify, and has wide application prospect for future large-scale medicine production and potential medical application thereof.
Drawings
FIG. 1 is an infrared spectrum of the first step reaction product PEG-PCL synthesized in example 2.
FIG. 2 is an infrared spectrum of the second reaction product PEG-PCL-PEG synthesized in example 2.
Fig. 3 is a graph showing the distribution of particle size of PECE-modified MA liposomes prepared in example 2.
FIG. 4 is a cold field scanning electron micrograph of PECE-modified MA liposomes prepared in example 2.
FIG. 5 is a graph comparing the PECE-modified MA liposomes prepared in example 2 at 4 ℃ and 32 ℃.
Fig. 6 shows the therapeutic effect of PECE-modified MA liposomes prepared in example 2 in scald experiments.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto. It is expressly intended that all such similar substitutes and modifications which would be obvious to those skilled in the art are deemed to be included within the invention.
The main reagents used in the embodiment of the invention are as follows:
polyethylene glycol monomethyl ether (MPEG, Mn 550, TCL, analytical grade)
Caprolactone (ε -CL, Macklin, analytical purity)
Stannous octoate (Sn (Oct)2Saen chemical technology Shanghai Co., Ltd.)
Hexamethylene diisocyanate (HMDI, Macklin, analytical pure)
Example 1
(1) 5.05g of MPEG and 10.96g of ε -CL (n) were weighed out(MPEG)：n(ε-CL)1:10) into a round bottom flask, 0.08g of stannous octoate (Sn (Oct)2) After 12h of reaction at 120 ℃ 1.66g of HMDI (n) were added(ε-CL)：n(HMDI)The ratio of 10:1) is reacted for 6h at 80 ℃, after purification, PEG-PCL-PEG triblock Polymer (PECE) is obtained, and vacuum drying is carried out until the weight is constant for later use.
(2) Weighing PECE 0.50g, yolk lecithin 0.50g, cholesterol 0.10g, and VEDissolving 0.05mL of acetic ester in 10mL of dichloromethane to prepare an organic phase solution;
(3) dissolving 20mg/mL madecassoside solution in the organic phase solution prepared in the step (2), and stirring for 5min at 500r/min to form W/O emulsion;
(4) transferring the W/O emulsion in the step (3) into an eggplant-shaped bottle, and removing the organic solvent by reduced pressure evaporation through a rotary evaporator; drying the eggplant-shaped bottle in a vacuum drying oven, removing the residual organic solvent in the emulsion, performing ultrasonic treatment for 60s, filtering with 0.45 μm oily filter membrane to obtain PEG-PCL-PEG triblock copolymer modified madecassoside liposome solution, and storing at 4 deg.C.
Example 2
(1) 5.05g of MPEG and 10.96g of ε -CL (n) were weighed out(MPEG)：n(ε-CL)1:10) into a round bottom flask, 0.08g of stannous octoate (Sn (Oct)2) After 12h of reaction at 120 ℃ 1.66g of HMDI (n) were added(ε-CL)：n(HMDI)The ratio of 10:1) is reacted for 6h at 80 ℃, and after purification, the mixture is dried in vacuum to constant weight for later use.
And respectively carrying out infrared spectrum characterization on the PEG-PCL as the first step reaction product and the PEG-PCL-PEG as the second step reaction product. As a result, as shown in FIGS. 1 and 2, it is understood from FIG. 1 that a strong carbonyl absorption peak appears in the 1700 to 1750 wavelength band, which means that an ester bond is formed by the reaction between ε -CL and MPEG; as can be seen from FIG. 2, no absorption peak appears between 2250 and 2270, which indicates that HMDI completely participates in the reaction to generate PEG-PCL-PEG, and a characteristic absorption peak of amide bond appears, further proving the synthesis of PEG-PCL-PEG.
(2) Weighing PECE 0.50g, yolk lecithin 0.50g, cholesterol 0.10g, and VEDissolving 0.05mL of acetic ester in 10mL of dichloromethane to prepare an organic phase solution;
(3) dissolving 20mg/mL MA solution in the organic phase solution prepared in the step (2), and stirring at 500r/min for 5min to form W/O emulsion; then adding the W/O emulsion into 20mg/mL MA PBS solution, and stirring for 5min at 500r/min for the second time to form W/O/W multiple emulsion;
(4) transferring the W/O/W multiple emulsion in the step (3) into an eggplant-shaped bottle, and removing the organic solvent by reduced pressure evaporation through a rotary evaporator; drying the eggplant-shaped bottle in a vacuum drying oven, removing the residual organic solvent in the emulsion, performing ultrasonic treatment for 60s, filtering with a 0.45-micrometer aqueous filter membrane to obtain PEG-PCL-PEG triblock copolymer modified madecassoside liposome solution, and storing at 4 deg.C.
(5) Carrying out particle size distribution test on the sample prepared in the step (4) by using a Malvern particle size analyzer, observing the apparent morphology by using a cold field Scanning Electron Microscope (SEM) and exploring the phase transition temperature;
the distribution graph of the sample particle size is shown in FIG. 3, and it can be seen that the PECE-modified MA liposomes have relatively uniform particle sizes, and the particle sizes are about 200 nm.
The cold field scanning electron micrograph of the sample is shown in fig. 4, and it can be seen from the micrograph that the PECE-modified MA liposomes have dispersed spherical shapes.
A comparison of the conditions of the samples at 4 ℃ and 32 ℃ is shown in FIG. 5, which shows that PECE-modified MA liposomes are in a fluid state at 4 ℃ and in a semi-pasty solid state at 32 ℃.
(6) 8 SD rats are taken and raised in a single cage, are fed with water freely, and are lighted for 12 hours and dark for 12 hours at room temperature. After all rats were anesthetized by intraperitoneal injection with sodium pentobarbital (40mg/kg), the dorsal rat hairs were mechanically shaved off, and the remaining rat hairs were treated with 8% Na2The S solution is removed. After 24h, a secondary scald model was created and randomly divided into PBS solution group, PECE drug-loaded group, MA liposome group and PECE modified MA liposome group, each group consisting of 2 rats, each rat having 6 wounds on the back. Rats in each group were dosed 2 times a day and wound healing was observed and recorded. The therapeutic effect of PECE-modified MA liposome in scald experiments is shown in fig. 6, and it can be known from the figure that the cure index of PECE-modified MA liposome is higher than that of MA liposome in the whole wound healing period, and the cure effect is better, and the PECE-modified MA liposome is a PECE drug-carrying agent and a PBS solution group.
Example 3
(1) Weighing 2.14g MPEG and 6.67g ε -CL (n)(MPEG)：n(ε-CL)1:15) into a round bottom flask, 0.05g of stannous octoate (Sn (Oct)2) After 15h of reaction at 100 ℃ 0.65g of HMDI (n) was added(ε-CL)：n(HMDI)The ratio of 15:1) is reacted for 9h at 50 ℃, and after purification, the mixture is dried in vacuum to constant weight for later use.
(2) Weighing 0.40g PECE, 0.50g hydrogenated soybean lecithin (HSPC), 0.10g cholesterol and 0.05mL BHA, and dissolving in 10mL chloroform to obtain organic phase solution;
(3) dissolving 160mg/mL MA solution in the organic phase solution prepared in the step (2), and stirring at 800r/min for 3min to form W/O emulsion; adding the W/O emulsion into 160mg/mL MA water solution, and stirring for 3min at 800r/min for the second time to form W/O/W multiple emulsion;
(4) transferring the W/O/W multiple emulsion in the step (3) into an eggplant-shaped bottle, and removing the organic solvent by reduced pressure evaporation through a rotary evaporator; drying the eggplant-shaped bottle in a vacuum drying oven, removing the residual organic solvent in the emulsion, performing ultrasonic treatment for 60s, filtering with a 0.45-micrometer aqueous filter membrane to obtain PEG-PCL-PEG triblock copolymer modified madecassoside liposome solution, and storing at 4 deg.C.
Example 4
(1) 3.46g of MPEG and 3.59g of ε -CL were weighed into a round bottom flask (n)(MPEG)：n(ε-CL)1:5), 0.04g stannous octoate (Sn (Oct)2) After 12h of reaction at 120 ℃ 5.29g of HMDI (n) were added(ε-CL)：n(HMDI)The ratio of 1:1) is reacted for 8h at 60 ℃, and after purification, the mixture is dried in vacuum to constant weight for later use.
(2) Weighing PECE 0.40g, yolk lecithin 0.50g, cholesterol 0.10g, and VEDissolving 0.05mL of acetic ester in 10mL of dichloromethane to prepare an organic phase solution;
(3) dissolving 160mg/mL madecassoside solution in the organic phase solution prepared in the step (2), and stirring at 400r/min for 10min to form a W/O emulsion;
(4) transferring the W/O emulsion in the step (3) into an eggplant-shaped bottle, and removing the organic solvent by reduced pressure evaporation through a rotary evaporator; drying the eggplant-shaped bottle in a vacuum drying oven, removing the residual organic solvent in the emulsion, performing ultrasonic treatment for 60s, filtering with 0.45 μm oily filter membrane to obtain PEG-PCL-PEG triblock copolymer modified madecassoside liposome solution, and storing at 4 deg.C.
Example 5
(1) Weighing 2.68g MPEG and 4.45g ε -CL (n)(MPEG)：n(ε-CL)1:8) into a round bottom flask, 0.04g stannous octoate (Sn (Oct)2) After 8h of reaction at 180 ℃ 1.64g of HMDI (n) were added(ε-CL)：n(HMDI)The ratio of 4:1) is reacted for 3h at 100 ℃, and after purification, the mixture is dried in vacuum to constant weight for later use.
(2) Weighing 0.40g of PECE, 0.50g of hydrogenated soybean lecithin (HSPC), 0.10g of cholesterol and 0.05mL of vitamin C, and dissolving in 10mL of diethyl ether to prepare an organic phase solution;
(3) dissolving 5mg/mL madecassoside solution in the organic phase solution prepared in the step (2), and stirring at 500r/min for 5min to form W/O emulsion; then adding the W/O emulsion into 5mg/mL MA PBS solution, and stirring for 5min at 500r/min for the second time to form W/O/W multiple emulsion;
(4) transferring the W/O/W multiple emulsion in the step (3) into an eggplant-shaped bottle, and removing the organic solvent by reduced pressure evaporation through a rotary evaporator; drying the eggplant-shaped bottle in a vacuum drying oven, removing the residual organic solvent in the emulsion, performing ultrasonic treatment for 60s, filtering with a 0.45-micrometer aqueous filter membrane to obtain PEG-PCL-PEG triblock copolymer modified madecassoside liposome solution, and storing at 4 deg.C.
Example 6
(1) Weigh 2.52g MPEG and 6.27g ε -CL (n)(MPEG)：n(ε-CL)1:12) into a round bottom flask, 0.05g of stannous octoate (Sn (Oct)2) After 4h of reaction at 200 ℃ 0.65g of HMDI (n) was added(ε-CL)：n(HMDI)The ratio of 14:1) is reacted for 9h at 50 ℃, and after purification, the mixture is dried in vacuum to constant weight for standby.
(2) Weighing 0.50g of PECE, 0.50g of soybean lecithin, 0.10g of cholesterol and 0.05mL of vitamin C, and dissolving in 10mL of trichloromethane to prepare an organic phase solution;
(3) dissolving 40mg/mL madecassoside solution in the organic phase solution prepared in the step (2), and stirring for 1min at 1000r/min to form a W/O emulsion;
(4) transferring the W/O emulsion in the step (3) into an eggplant-shaped bottle, and removing the organic solvent by reduced pressure evaporation through a rotary evaporator; drying the eggplant-shaped bottle in a vacuum drying oven, removing the residual organic solvent in the emulsion, performing ultrasonic treatment for 60s, filtering with 0.45 μm oily filter membrane to obtain PEG-PCL-PEG triblock copolymer modified madecassoside liposome solution, and storing at 4 deg.C.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (10)

1. A PEG-PCL-PEG triblock copolymer modified madecassoside liposome is characterized in that the liposome comprises a PEG-PCL-PEG triblock copolymer, madecassoside, cholesterol and lipid components; the madecassoside is encapsulated in a liposome;
the lipid component is phospholipid;
the preparation method of the PEG-PCL-PEG triblock copolymer comprises the following steps:
adding caprolactone and polyethylene glycol monomethyl ether, under the action of a catalyst, breaking ester bonds in epsilon-CL, reacting with hydroxyl in MPEG to generate new ester bonds, namely PEG-PCL fragments, namely, reacting in the first step, then adding hexamethylene diisocyanate, continuously reacting isocyanate groups in the ester bonds with the hydroxyl generated in the PEG-PCL to generate amide bonds, namely, reacting in the second step, purifying to obtain a PEG-PCL-PEG triblock polymer, and drying in vacuum to constant weight for later use;
the preparation method of the PEG-PCL-PEG triblock copolymer modified madecassoside liposome comprises the following steps:
(1) weighing PEG-PCL-PEG triblock polymer, lipid component, cholesterol and a small amount of antioxidant, and dissolving in a certain amount of organic solvent to obtain organic phase solution;
(2) adding the aqueous phase solution containing the madecassoside into the organic phase solution obtained in the step (1), and stirring to form a W/O emulsion, or adding the W/O emulsion into a second aqueous phase, and stirring to form a W/O/W multiple emulsion;
(3) and (3) evaporating the W/O emulsion or W/O/W multiple emulsion in the step (2) under reduced pressure to remove the organic solvent, then carrying out vacuum drying to remove the residual organic solvent in the emulsion, and filtering the membrane to obtain the PEG-PCL-PEG triblock copolymer modified madecassoside liposome.
2. The method for preparing PEG-PCL-PEG triblock copolymer modified madecassoside liposome of claim 1, characterized by comprising the following steps:
(1) weighing PEG-PCL-PEG triblock polymer, lipid component, cholesterol and a small amount of antioxidant, and dissolving in a certain amount of organic solvent to obtain organic phase solution;
(2) adding the aqueous phase solution containing the madecassoside into the organic phase solution obtained in the step (1), and stirring to form a W/O emulsion, or adding the W/O emulsion into a second aqueous phase, and stirring to form a W/O/W multiple emulsion;
(3) and (3) evaporating the W/O emulsion or W/O/W multiple emulsion in the step (2) under reduced pressure to remove the organic solvent, then carrying out vacuum drying to remove the residual organic solvent in the emulsion, and filtering the membrane to obtain the PEG-PCL-PEG triblock copolymer modified madecassoside liposome.
3. The method for preparing the PEG-PCL-PEG triblock copolymer modified madecassoside liposome according to claim 2, wherein the method comprises the following steps:
in the step (1), the mass ratio of the PEG-PCL-PEG triblock polymer to the lipid component is (0.5-1): 1;
the mass ratio of the lipid component to the cholesterol is 5: 1;
the concentration of cholesterol in the organic phase solution in the step (1) is 10-50 mg/mL.
4. The method for preparing the PEG-PCL-PEG triblock copolymer modified madecassoside liposome according to claim 2, wherein the method comprises the following steps:
the antioxidant is vitamin C, BHA and vitamin E acetate;
the organic solvent is diethyl ether, dichloromethane or trichloromethane.
5. The method for preparing the PEG-PCL-PEG triblock copolymer modified madecassoside liposome according to claim 2, wherein the method comprises the following steps:
in the step (2), the water phase or the second water phase is an aqueous solution or a PBS solution containing madecassoside, and the concentration is 1-160 mg/mL; stirring for 1-30 min; stirring at a speed of 100-1000 rpm; stirring for the second time for 1-30 min by a multiple emulsion method; the stirring speed is 100-1000 rpm.
6. The method for preparing the PEG-PCL-PEG triblock copolymer modified madecassoside liposome according to claim 2, wherein the method comprises the following steps:
in the step (3), the aperture of the filter membrane is 0.22-1 μm; the madecassoside liposome prepared by W/O emulsion is applied to an oily filter membrane, and the MA liposome prepared by W/O/W multiple emulsion is applied to an aqueous filter membrane.
7. The method for preparing the PEG-PCL-PEG triblock copolymer modified madecassoside liposome according to claim 2, wherein the method comprises the following steps:
the preparation method of the PEG-PCL-PEG triblock copolymer in the step (1) comprises the following steps:
adding caprolactone and polyethylene glycol monomethyl ether, under the action of a catalyst, breaking ester bonds in epsilon-CL, reacting with hydroxyl in MPEG to generate new ester bonds, namely PEG-PCL fragments, namely, reacting in the first step, then adding hexamethylene diisocyanate, continuously reacting isocyanate groups in the ethylene glycol-CL with the hydroxyl generated in the PEG-PCL to generate amide bonds, namely, reacting in the second step, purifying to obtain a PEG-PCL-PEG triblock polymer, and drying in vacuum to constant weight for later use.
8. The method for preparing the PEG-PCL-PEG triblock copolymer modified madecassoside liposome according to claim 7, wherein the method comprises the following steps:
first step reaction raw material ratio n(MPEG)：n(ε-CL)Is 1: (1-15); the reaction conditions are that the reaction is carried out for 4 to 15 hours at the temperature of 100 to 200 ℃;
the ratio of raw materials in the second step n(ε-CL)：n(HMDI)The ratio of (1-15): 1; the reaction conditions are 50-100 ℃ for 3-9 h.
9. The PEG-PCL-PEG triblock copolymer modified madecassoside liposome of claim 1, which is applied to the technical field of pharmaceutical preparations or skin care cosmetics.
10. Use according to claim 9, characterized in that:
the application of the madecassoside liposome in preparing a transdermal or mucosal absorption pharmaceutical preparation for treating skin scald, skin cut, mucosal injury or scar;
the application of the madecassoside liposome in preparing skin care cosmetics for scar repair, deep skin damage or anti-aging, anti-oxidation or whitening and beautifying.
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CN1430952A (en) * 2003-01-30 2003-07-23 上海家化联合股份有限公司 Asiaticoside liposome and its use
WO2013075846A1 (en) * 2011-11-23 2013-05-30 Pls Design Gmbh Pharmaceutical composition for treatment of allergic reactions
CN103893122A (en) * 2014-03-28 2014-07-02 华南理工大学 Madecassoside lipidosome and preparation method and application thereof
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CN1430952A (en) * 2003-01-30 2003-07-23 上海家化联合股份有限公司 Asiaticoside liposome and its use
WO2013075846A1 (en) * 2011-11-23 2013-05-30 Pls Design Gmbh Pharmaceutical composition for treatment of allergic reactions
EP2918262A1 (en) * 2014-03-10 2015-09-16 PLS-Design GmbH Induction of antigen-specific tolerance by peripheral phagocytosis
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