CN116115826A

CN116115826A - Polycaprolactone gel filling material coated with geniposide acid and preparation method thereof

Info

Publication number: CN116115826A
Application number: CN202310205283.2A
Authority: CN
Inventors: 雷建都; 王洪宇
Original assignee: Beijing Forestry University
Current assignee: Beijing Forestry University
Priority date: 2023-03-06
Filing date: 2023-03-06
Publication date: 2023-05-16

Abstract

The invention discloses a polycaprolactone gel coated with geniposide acid, which comprises a filling material and a preparation method thereof and is characterized in that the geniposide acid is coated in polycaprolactone microspheres and dispersed in a gel matrix to obtain gel for injection, and the gel is applied to facial filling. The invention has simple process and good repeatability, and the polycaprolactone filling material prepared by the invention has good facial filling effect and anti-aging effect, and is a novel facial filling material with excellent performance and good application prospect in the field of medical cosmetology.

Description

Polycaprolactone gel filling material coated with geniposide acid and preparation method thereof

Technical Field

The invention relates to a polycaprolactone gel filling material for encapsulating geniposide acid and a preparation method thereof, belonging to the technical field of medical cosmetology.

Background

As the human body ages, facial collagen fibers and elastic fibers gradually decrease, leading to skin relaxation and reduced elasticity, which is one of the most obvious signs of facial aging. The initial method for improving facial aging is to simply remove loose skin by surgery, and although facial relaxation can be effectively improved, there are also some disadvantages such as obvious incision scars, slow recovery, etc. With the continuous development and progress of technology, facial rejuvenation gradually progresses from pure surgical treatment to the application of various methods such as surgery, laser, injection filling and the like. At present, the face injection filling treatment is a minimally invasive treatment with lighter damage to human bodies and shorter recovery time, and can better make up for the face defects caused by aging, so that in the increasing demand of face rejuvenation treatment, the popularity of the face injection filling material in the past 20 years is exponentially increased.

The goal of facial filling therapy is to rebuild the atrophic subcutaneous fat and dermis to restore it to a younger state. The facial filling is mainly used for improving facial soft tissue defects, skin static wrinkles and tissue contours, and the ideal facial filling not only has attractive and durable effects, but also has good biocompatibility and safety.

The initial face pack is an animal-derived collagen pack, but has been rarely applied to face pack in recent years due to the disadvantages of skin hypersensitivity test before treatment and short duration of effect after treatment. Compared with animal collagen, hyaluronic Acid (HA) is commonly called Hyaluronic acid, is a main component of extracellular matrix, is widely existing in human skin, HAs non-immunogenicity, and HAs less allergic reaction. Hyaluronic acid is used as a multifunctional matrix in the organism, and has important physiological functions of regulating and controlling cell proliferation and differentiation, promoting wound healing, resisting oxidation, resisting aging and the like. Of particular importance is its good viscoelasticity and moisture retention, which is currently one of the most commonly used facial filling materials. The metabolism time of uncrosslinked hyaluronic acid in vivo is only about 1 to 2 days, and recent studies have revealed that the particle size and molecular weight can be increased by crosslinking reaction to make it a gel-like polymer which is not easily enzymatically hydrolyzed by endogenous hyaluronidase, thereby extending the filling time in vivo. When crosslinked hyaluronic acid is used as a raw material for skin fillers, repeated treatments every 6 to 12 months are required, although the sustained effect is prolonged to some extent. In addition, hyaluronic acid has no other effects than physical support and water absorption, and cannot meet higher anti-aging requirements.

In recent years, face fillers made of synthetic high molecular polymers have been attracting attention due to their good properties. The synthetic high molecular polymer is very slowly decomposed in the human body, and among them, polycaprolactone (PCL) is a good facial filling material, and studies have shown that Polycaprolactone having a number average molecular weight of 50kDa takes three years to be completely degraded from the body, and thus has a longer duration of facial filling effect than collagen and hyaluronic acid fillers.

The polycaprolactone is a degradable synthetic high polymer material formed by ring-opening polymerization of epsilon-caprolactone under the catalysis of a metal anion complex catalyst, and different molecular weights can be obtained by controlling polymerization conditions, and the basic structural unit of the polycaprolactone is shown as a formula I.

It is insoluble in water, but soluble in various organic solvents, and can be completely degraded into carbon dioxide and water in natural environment, and the degradation products can be absorbed by body or discharged from body, so that it has good biocompatibility and degradability, and can be extensively used in the fields of biomedical engineering, medicine and health, medical cosmetology, etc. Since the 80 s of the 20 th century, polycaprolactone was applied in the biomedical field, and most of the latest applications are to prepare the polycaprolactone into microspheres to be used as facial fillers, and the prepared gel can not only provide facial physical support, but also stimulate skin collagen regeneration, improve facial relaxation and tissue contour, and has better facial rejuvenation efficacy.

Geniposidic acid (GPA) is an iridoid compound extracted and purified from plants such as gardenia, semen plantaginis, eucommia ulmoides and the like, and has good anti-aging property, so that the Geniposidic acid (GPA) is applied to face rejuvenation treatment, and the basic structural unit is shown as a formula II.

Geniposide has not only antioxidant and anti-aging pharmacological effects, but also good biocompatibility and degradability, and has been studied by taking rats as study objects, which proves that geniposide can promote proliferation of rat epidermal cells and synthesis of collagen, and has obvious effect of relieving aging. Although geniposide has remarkable antioxidant and antiaging effects, geniposide has poor stability in human body and is easily decomposed, thereby affecting the pharmacological effect and being limited in application.

Therefore, there is a need to develop a new polycaprolactone gel filler having improved efficacy by solving the in vivo stability problem of geniposide acid by entrapping geniposide acid with polycaprolactone microspheres, and dispersing the microspheres in a gel matrix to make a geniposide acid entrapped polycaprolactone gel for injection.

Disclosure of Invention

The invention aims to provide a polycaprolactone gel filling material coated with geniposide acid and a preparation method thereof, and aims to obtain polycaprolactone gel for injection with good facial filling effect and anti-aging effect.

In order to achieve the above object, according to a first aspect of the present invention, there is provided a method for preparing a polycaprolactone microsphere coated with geniposide acid. The method is realized by the following steps:

dissolving polycaprolactone in an organic solvent I, dissolving geniposide acid in an organic solvent II, and uniformly mixing the two solutions to prepare a single solution serving as an oil phase; preparing an emulsion by mixing the above oil phase with a surfactant-containing dispersion (aqueous phase); extracting the organic solvent from the oil phase to the water phase by stirring the emulsion, evaporating the organic solvent for a certain time to solidify the liquid drops to form microspheres, collecting the microspheres from the water phase, and drying the microspheres in vacuum to obtain the polycaprolactone microspheres coated with geniposide acid.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the number average molecular weight of polycaprolactone is 5-100 kDa.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the organic solvent I for dissolving polycaprolactone is one or a combination of more of dichloromethane, ethyl acetate and cyclohexane.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the organic solvent II for dissolving geniposide acid is one or a combination of more of dimethyl sulfoxide, methanol and ethanol.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the surfactant is one or a combination of more of polyvinylpyrrolidone, carboxymethyl cellulose, polyvinyl alcohol and span.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the content of the selected surfactant in the water phase can be 0.5-5 w/v% of the total volume of the water phase.

In the preparation method of the geniposide-entrapped polycaprolactone microsphere, preferably, the selected aqueous dispersion is one of purified water, normal saline and phosphate buffer. Specifically, the phosphate buffer solution is coated with one or more of the combination of the hydrogen phosphate and the dihydrogen phosphate, and the pH value is 6.0-8.0. Preferably, the selected dibasic phosphate is one or more of disodium hydrogen phosphate, dipotassium hydrogen phosphate and diammonium hydrogen phosphate, and the selected monobasic phosphate is one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate or monoammonium dihydrogen phosphate.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, an emulsion is prepared by mixing an oil phase and a water phase by adopting a method such as an emulsion volatilization method, a phase separation method, a spray drying method, a microfluidic method, a membrane emulsification method and the like.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the stirring speed is selected to be 100-4000 rpm, and the time for evaporating the organic solvent from the water phase by stirring is 0.5-24 h.

In the preparation method of the polycaprolactone microsphere coated with geniposide acid, preferably, the method of collecting the microsphere from the water phase is one or a combination of filtration and centrifugation.

In the preparation method of the polycaprolactone microsphere coated with geniposide, preferably, the content of the geniposide in the polycaprolactone microsphere is 0.1-20 w/w%.

In the preparation method of the geniposide-entrapped polycaprolactone microsphere, preferably, the average granularity of the selected geniposide-entrapped polycaprolactone microsphere is 1-100 μm.

According to a second aspect of the present invention, there is provided a method for preparing a dispersion matrix of polycaprolactone microspheres entrapped geniposide acid, by: and uniformly dispersing the prepared dried polycaprolactone microsphere coated with geniposide acid into a dispersion liquid to obtain a dispersion matrix.

In the preparation method of the dispersing matrix of the polycaprolactone microsphere coated with geniposide acid, preferably, the dispersing liquid is selected as follows: purified water, physiological saline and phosphate buffer. Specifically, it is defined by the dispersion according to the first aspect of the invention.

According to a third aspect of the present invention, there is provided a preparation method of a gel for injection of polycaprolactone entrapping geniposide acid, which is implemented by the following steps: adding solute into the dispersion liquid, mixing to obtain gel matrix, and mixing the prepared dispersion matrix, gel matrix and glycerin to obtain gel for injection.

In the preparation method of the gel for injection of the polycaprolactone coated with geniposide acid, preferably, the solute is selected from one or more of carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, hyaluronic acid and lidocaine.

In the preparation method of the gel for injection of the polycaprolactone coated with geniposide acid, preferably, the dispersion liquid is selected as follows: purified water, physiological saline and phosphate buffer. Specifically, it is defined by the dispersion according to the first aspect of the invention.

According to the invention, through regulating and controlling the process of the polycaprolactone-entrapped geniposide acid, the filling material with good facial tissue repair characteristics is prepared, so that the in-vivo stability of the geniposide acid is improved, good anti-aging characteristics are exerted, the duration of facial filling effect is prolonged, and the filling material has a better facial rejuvenation effect.

Drawings

FIG. 1 is an optical micrograph of a geniposide-entrapped polycaprolactone microsphere;

FIG. 2 is a scanning electron micrograph of geniposide acid-entrapped polycaprolactone microspheres prepared according to example 1-1;

FIG. 3 is a scanning electron micrograph of geniposide acid entrapped polycaprolactone microspheres prepared according to examples 1-2;

fig. 4 is a photograph of the appearance of a geniposide entrapped polycaprolactone emulsion.

Detailed Description

The examples given below illustrate the invention in detail, without limiting it, the scope of which is defined by the claims.

Example 1:

preparing a polycaprolactone microsphere coated with geniposide acid: 10g of polycaprolactone with Mn of 80k is dissolved in 100mL of dichloromethane, 100mg of geniposide acid is dissolved in 10mL of dimethylsulfoxide, and the two solutions are uniformly mixed by ultrasound to prepare a single solution as an oil phase; adding water to 0.2mol/L disodium hydrogen phosphate solution and 0.2mol/L sodium dihydrogen phosphate solution for dilution to prepare phosphate buffer solution with pH of 7.2, and dissolving 10g of polyvinyl alcohol in 1000mL of phosphate buffer solution to serve as a water phase; preparing emulsion by stirring and emulsifying the oil phase and the water phase through an emulsion volatilization method, and stirring at a speed of 200rpm for 5 hours, thereby evaporating and removing the organic solvent and solidifying the liquid drops to form microspheres; repeatedly washing and centrifuging the microsphere suspension with water, and freeze-drying the obtained microsphere to obtain the dried polycaprolactone microsphere coated with geniposide acid.

Preparation of a dispersion matrix of microspheres: 10g of the prepared dried geniposide-entrapped polycaprolactone microsphere was uniformly dispersed into 10mL of the prepared phosphate buffer solution to obtain a dispersion matrix.

Preparation of gel for injection: mixing 1g of carboxymethyl cellulose with 20mL of phosphate buffer solution prepared above to obtain a gel matrix; then, the dispersion matrix prepared above, the gel matrix and 0.5g of glycerin were mixed to obtain a gel for injection.

Example 2:

preparing a polycaprolactone microsphere coated with geniposide acid: 10g of polycaprolactone with Mn of 40k was dissolved in 100mL of dichloromethane, 50mg of geniposide acid was dissolved in 8mL of dimethylsulfoxide, and the two solutions were uniformly mixed by ultrasound to prepare a single solution as an oil phase; adding water to 0.2mol/L disodium hydrogen phosphate solution and 0.2mol/L sodium dihydrogen phosphate solution for dilution to prepare phosphate buffer solution with pH of 7.0, and dissolving 15g of polyvinyl alcohol in 1000mL of phosphate buffer solution to serve as a water phase; preparing emulsion by stirring and emulsifying the oil phase and the water phase through an emulsion volatilization method, and stirring at a speed of 150rpm for 3 hours, thereby evaporating and removing the organic solvent and solidifying the liquid drops to form microspheres; repeatedly washing and centrifuging the microsphere suspension with water, and freeze-drying the obtained microsphere to obtain the dried polycaprolactone microsphere coated with geniposide acid.

Preparation of gel for injection: mixing 1g of carboxymethyl cellulose with 10mL of phosphate buffer solution prepared above to obtain a gel matrix; then, the dispersion matrix prepared above, the gel matrix and 0.5g of glycerin were mixed to obtain a gel for injection.

Example 3:

preparing a polycaprolactone microsphere coated with geniposide acid: 10g of polycaprolactone with Mn of 60k is dissolved in 80mL of ethyl acetate, 150mg of geniposide acid is dissolved in 10mL of methanol, and the two solutions are uniformly mixed by ultrasonic to prepare a single solution as an oil phase; adding water to 0.2mol/L dipotassium hydrogen phosphate solution and 0.2mol/L potassium dihydrogen phosphate solution to dilute to prepare a phosphate buffer solution with pH of 6.8, and dissolving 20g span in 1000mL of the phosphate buffer solution to serve as a water phase; preparing emulsion by stirring and emulsifying the oil phase and the water phase through an emulsion volatilization method, and stirring at a speed of 300rpm for 8 hours, thereby evaporating and removing the organic solvent and solidifying the liquid drops to form microspheres; repeatedly washing and centrifuging the microsphere suspension with water, and freeze-drying the obtained microsphere to obtain the dried polycaprolactone microsphere coated with geniposide acid.

Preparation of gel for injection: mixing 0.5g of sodium carboxymethyl cellulose with 5mL of phosphate buffer solution prepared above to obtain a gel matrix; the dispersion matrix prepared above, the gel matrix and 0.25g of glycerin were then mixed to obtain a gel for injection.

Example 4:

preparing a polycaprolactone microsphere coated with geniposide acid: 10g of polycaprolactone with Mn of 50k is dissolved in 150mL of ethyl acetate, 100mg of geniposide acid is dissolved in 15mL of methanol, and the two solutions are uniformly mixed by ultrasonic to prepare a single solution as an oil phase; adding water to 0.1mol/L disodium hydrogen phosphate solution and 0.1mol/L potassium dihydrogen phosphate solution for dilution to prepare a phosphate buffer solution with pH of 6.5, and dissolving 15g span in 1000mL of the phosphate buffer solution to serve as a water phase; preparing emulsion by stirring and emulsifying the oil phase and the water phase through an emulsion volatilization method, and stirring at 400rpm for 10 hours, thereby evaporating and removing the organic solvent and solidifying the liquid drops to form microspheres; repeatedly washing and centrifuging the microsphere suspension with water, and freeze-drying the obtained microsphere to obtain the dried polycaprolactone microsphere coated with geniposide acid.

Preparation of a dispersion matrix of microspheres: 10g of the prepared dried geniposide-entrapped polycaprolactone microsphere was uniformly dispersed in 15mL of the prepared phosphate buffer solution to obtain a dispersion matrix.

Preparation of gel for injection: mixing 1g of sodium carboxymethyl cellulose with 10mL of phosphate buffer solution prepared above to obtain a gel matrix; then, the dispersion matrix prepared above, the gel matrix and 0.5g of glycerin were mixed to obtain a gel for injection.

Example 5:

preparing a polycaprolactone microsphere coated with geniposide acid: 10g of polycaprolactone with Mn of 80k is dissolved in 100mL of cyclohexane, 200mg of geniposide acid is dissolved in 20mL of methanol, and the two solutions are uniformly mixed by ultrasonic to prepare a single solution as an oil phase; adding water to 0.1mol/L disodium hydrogen phosphate solution and 0.1mol/L potassium dihydrogen phosphate solution to dilute to obtain phosphate buffer solution with pH of 7.4, and dissolving 10g polyvinylpyrrolidone in 800mL phosphate buffer solution to obtain water phase; preparing emulsion by stirring and emulsifying the oil phase and the water phase through an emulsion volatilization method, and stirring at a speed of 200rpm for 15 hours, thereby evaporating and removing the organic solvent and solidifying the liquid drops to form microspheres; repeatedly washing and centrifuging the microsphere suspension with water, and freeze-drying the obtained microsphere to obtain the dried polycaprolactone microsphere coated with geniposide acid.

Preparation of a dispersion matrix of microspheres: 10g of the prepared dried geniposide-entrapped polycaprolactone microsphere was uniformly dispersed in 18mL of the prepared phosphate buffer solution to obtain a dispersion matrix.

Preparation of gel for injection: mixing 2g of sodium carboxymethyl cellulose with 20mL of phosphate buffer solution prepared above to obtain a gel matrix; then, the dispersion matrix prepared above, the gel matrix and 1g of glycerin were mixed to obtain a gel for injection.

Example 6:

preparing a polycaprolactone microsphere coated with geniposide acid: 10g of polycaprolactone with Mn of 100k is dissolved in 120mL of cyclohexane, 150mg of geniposide acid is dissolved in 15mL of methanol, and the two solutions are uniformly mixed by ultrasonic to prepare a single solution as an oil phase; adding water to 0.2mol/L disodium hydrogen phosphate solution and 0.2mol/L potassium dihydrogen phosphate solution to dilute to obtain phosphate buffer solution with pH of 7.0, and dissolving 15g polyvinylpyrrolidone in 1200mL phosphate buffer solution to obtain water phase; preparing emulsion by stirring and emulsifying the oil phase and the water phase through an emulsion volatilization method, and stirring at a speed of 150rpm for 12 hours, thereby evaporating and removing the organic solvent and solidifying the liquid drops to form microspheres; repeatedly washing and centrifuging the microsphere suspension with water, and freeze-drying the obtained microsphere to obtain the dried polycaprolactone microsphere coated with geniposide acid.

Preparation of a dispersion matrix of microspheres: 10g of the prepared dried geniposide-entrapped polycaprolactone microsphere was uniformly dispersed into 20mL of the prepared phosphate buffer solution to obtain a dispersion matrix.

Preparation of gel for injection: mixing 1g of hydroxypropyl methylcellulose with 10mL of phosphate buffer prepared above to obtain a gel matrix; then, the dispersion matrix prepared above, the gel matrix and 0.5g of glycerin were mixed to obtain a gel for injection.

Claims

1. A preparation method of a polycaprolactone gel filling material coated with geniposide acid comprises the following steps:

(1) Adding solute into the dispersion liquid, and mixing to obtain gel matrix;

(2) Mixing the dispersion matrix, the gel matrix and the glycerol to obtain the gel filling material.

2. The method for preparing the geniposide-entrapped polycaprolactone gel filler according to claim 1, wherein the solute is one or more of carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, hyaluronic acid, and lidocaine.

3. The method for preparing the geniposide-entrapped polycaprolactone gel filling material according to claim 1, wherein the dispersion liquid is one of purified water, physiological saline and phosphate buffer.

4. The method for preparing the geniposide-entrapped polycaprolactone gel filler according to claim 1, wherein the dispersion matrix is a uniformly dispersed dispersion of geniposide-entrapped polycaprolactone microspheres.

5. The preparation method of the geniposide-entrapped polycaprolactone microsphere according to claim 4, which is characterized by comprising the following steps:

(1) Dissolving polycaprolactone in an organic solvent I, dissolving geniposide acid in an organic solvent II, and uniformly mixing the two solutions to prepare a single solution serving as an oil phase;

(2) Preparing an emulsion by mixing the oil phase with a surfactant-containing dispersion (aqueous phase) and emulsifying the mixture;

(3) Extracting the organic solvent from the oil phase to the water phase by stirring the emulsion, evaporating the organic solvent for a certain time to solidify the liquid drops to form microspheres, collecting the microspheres from the water phase, and drying the microspheres in vacuum to obtain the polycaprolactone microspheres coated with geniposide acid.

6. The method for preparing the polycaprolactone microsphere coated with geniposide acid according to claim 5, wherein the organic solvent I for dissolving polycaprolactone is one or more of dichloromethane, ethyl acetate and cyclohexane.

7. The method for preparing the polycaprolactone microsphere coated with geniposide acid according to claim 5, wherein the organic solvent II for dissolving polycaprolactone is one or more of dimethyl sulfoxide, methanol and ethanol.