CN110559261A

CN110559261A - Liposome microemulsion containing nano cross-linked hyaluronic acid and preparation method and application thereof

Info

Publication number: CN110559261A
Application number: CN201810574806.XA
Authority: CN
Inventors: 何浩明; 杨一菲; 于一帆
Original assignee: CHANGZHOU INSTITUTE OF MATERIA MEDICA Co Ltd
Current assignee: CHANGZHOU INSTITUTE OF MATERIA MEDICA Co Ltd
Priority date: 2018-06-06
Filing date: 2018-06-06
Publication date: 2019-12-13
Also published as: WO2019233368A1

Abstract

The invention discloses a liposome microemulsion containing hyaluronic acid, a preparation method and an application thereof, wherein the liposome microemulsion consists of the following components in percentage by weight: 0.5 to 1.5 percent of phospholipid, 0.1 to 0.6 percent of cholesterol, 0.05 to 0.5 percent of nano cross-linked hyaluronic acid or sodium salt thereof, 0.1 to 0.6 percent of medicinal surfactant and the balance of water. The liposome solution of the present invention is used as a vehicle for ophthalmic preparations such as eye spray, which delivers eye nutrients to the surface of the eyeball while delivering lipid materials to the eyeball, thereby improving 'dry eye' caused by liposome disorders; and the moisturizing factor nano cross-linked sodium hyaluronate is added into the liposome solution, and the cross-linked sodium hyaluronate nanoparticles can reach the surface of the eyeball through the eyelid to play a role in moisturizing the eyeball.

Description

Liposome microemulsion containing nano cross-linked hyaluronic acid and preparation method and application thereof

Technical Field

The invention relates to a liposome microemulsion containing nano cross-linked hyaluronic acid, a preparation method and application thereof.

background

the primary means currently used to treat dry eye is the instillation of eye drops or artificial "tears", which unfortunately are not always effective. The reason for this is that "dry eye" is not caused by less tears, but rather by rapid loss of lipids due to disturbance of liposomes on the surface of the eyeball.

Dry eye is largely classified into water-deficient dry eye, evaporative dry eye, and mixed dry eye. Aqueous deficient dry eye is caused by insufficient tear secretion. The excessive evaporation type dry eye is caused by excessive tear loss from the exposed surface of the eyeball under the condition that the function of lacrimal secretion is normal, and the causes thereof include exogenous causes (such as ocular surface diseases caused by wearing contact lenses or deficiency of vitamin A), endogenous causes (such as meibomian gland dysfunction and eyelid closure disorder).

For over-evaporated dry eye and mixed dry eye, the effect of dropping eyedrops or artificial tears is not good, and the method for relieving symptoms is to supplement lipid.

As a product for treating dry eye, there is a "liposome eye spray" marketed in Germany under the trade name "tears agar" (A new therapy concept with a liposome eye spray for the treatment of the dry eye "(Germ) [ J ]. Dkgest of the World Latest Medical Information, 2005.). The spray has the advantages of good biocompatibility, no toxicity and no immunogenicity, and the curative effect of the spray is indeed better than that of common eye drops through clinical examination. However, in the prescription of the lipid experience spray, a moisturizing factor hyaluronic acid which is known to be effective for dry eye is lacked; tests prove that when hyaluronic acid is added into the formula, precipitation is generated, so that the whole liposome microemulsion system is unstable, and a commodity cannot be formed.

Chinese patent document CN 106459137a (application No. 201580019868.9) discloses a membrane-attached self-assembled system for treating eye disorders, the preparation method of the liquid crystal drug delivery system is as follows: forming a first solution comprising a lipid component and an alcohol, the first solution being maintained at a first temperature of 40-55 ℃; obtaining a second solution comprising a mucoadhesive hydrophilic polymer and a buffer, the second solution being aqueous and maintained at a second temperature of 5-55 ℃; mixing the first solution and the second solution to form a combined nano/micro-dispersion, the mixing being achieved by a high energy mixing process; microfluidization of the combined nano/micro-dispersion at a third temperature of 40-55 ℃ to form a nano-dispersion; and incubating the nano-dispersion at 2-5 ℃ to form a liquid crystalline drug delivery system. The liquid crystal drug delivery system may be a spray, an injection, or formulated as eye drops.

the liquid crystal drug delivery system may be a high viscosity liquid that is pre-loaded into a syringe. However, according to the patent document, the liquid crystal drug delivery system is not long enough in storage stability, can be maintained for only 3 to 6 months, cannot form a commercial product with a long shelf life, does not contain a moisturizing factor sodium hyaluronate in the components, and is not good in use value.

disclosure of Invention

The invention aims to solve the technical problem of providing a stable liposome solution containing hyaluronic acid with long storage period, a preparation method and application thereof.

the technical scheme for realizing the first purpose of the invention is that the liposome microemulsion consists of the following components in percentage by weight: 0.5 to 1.5 percent of phospholipid, 0.1 to 0.6 percent of cholesterol, 0.05 to 0.5 percent of nano cross-linked hyaluronic acid or sodium salt thereof, 0.1 to 0.6 percent of medicinal surfactant and the balance of water.

Wherein the particle size of the nano cross-linked hyaluronic acid or the sodium salt thereof is 50 nm-250 nm.

The medicinal surfactant is one of tween, span and poloxamer.

The second object of the present invention is achieved by a first preparation method of the liposome microemulsion, comprising the steps of:

Firstly, dissolving weighed phospholipid and cholesterol in ethanol to obtain a lipid solution, and heating and evaporating to remove ethanol to obtain a lipid membrane.

② mixing the nano cross-linked sodium hyaluronate gel with water as dispersant to obtain hydration liquid.

Mixing the lipid membrane with the hydration liquid to obtain a mixed solution; wherein the pharmaceutically acceptable surfactant is added to the lipid solution or the hydration solution before the lipid film is mixed with the hydration solution; preparing the mixed solution into microemulsion to obtain liposome solution containing hyaluronic acid.

the second preparation method of the liposome microemulsion as described above comprises the steps of:

Firstly, dissolving weighed phospholipid and cholesterol in ethanol to obtain a lipid solution.

Mixing the lipid solution with the hydration solution to obtain a mixed solution; wherein the pharmaceutically acceptable surfactant is added to the lipid solution or the aqueous solution prior to mixing the lipid solution with the aqueous solution; heating and evaporating to remove ethanol; and preparing the mixed solution after removing the alcohol into microemulsion to obtain the liposome solution containing the hyaluronic acid.

The technical scheme for realizing the third object of the invention is the application of the liposome microemulsion in preparing the ophthalmic preparation for treating or relieving dry eye syndrome, wherein the liposome solution is an effective component in the ophthalmic preparation and a carrier of an ophthalmic nutritional agent.

The ophthalmic preparation is an eye spray.

The eye spray consists of the following components in percentage by weight: 0.5 to 2.1 percent of phospholipid, 0.1 to 0.6 percent of cholesterol, 0.05 to 0.5 percent of nano cross-linked hyaluronic acid or sodium salt thereof, 0.1 to 0.6 percent of medicinal surfactant, 0.001 to 1 percent of eye nutrient and the balance of water.

Further, the eye nutrient is one or more of taurine, hypromellose, Va, Ve, Vc, lutein, zeaxanthin, beta-carotene, anthocyanin, DHA and vitamin B6.

A first method of preparing an eye spray as described above, comprising the steps of:

Dissolving weighed phospholipid, cholesterol and eye nutrient in ethanol, uniformly mixing, stirring at high speed, coating the eye nutrient with liposome, and heating to evaporate to remove ethanol to obtain lipid membrane.

A second method of preparing an eye spray as described above, comprising the steps of:

firstly, dissolving the weighed phospholipid, cholesterol and the eye nutrient in ethanol, uniformly mixing, stirring at a high speed, and coating the eye nutrient with liposome to obtain a lipid solution.

the invention has the positive effects that:

(1) The liposome solution of the present invention is used as a vehicle for ophthalmic preparations such as eye spray, which delivers eye nutrients to the surface of the eyeball while delivering lipid materials to the eyeball, thereby improving 'dry eye' caused by liposome disorders; in addition, the moisturizing factor nano cross-linked sodium hyaluronate is added into the liposome solution, and the cross-linked sodium hyaluronate nanoparticles can reach the surface of the eyeball through the eyelid to play a role in moisturizing the eyeball; in addition, the nano cross-linked sodium hyaluronate, the liposome microemulsion and ocular nutrients such as Va, Ve and the like wrapped by the microemulsion act together to achieve the effects of moistening and nourishing eyes, so that the whole eyes can achieve the effects of moistening, relieving and improving eyesight.

(2) The eye spray disclosed by the invention has the effect of treating dry eye, wherein Va and Ve added into the eye spray can also increase the elasticity of eye skin; the added cross-linked hyaluronic acid can more effectively smooth fine lines, prevent wrinkles, delay skin aging around eyes, and tighten eye bags, so that the eye skin becomes soft, tender and elastic after long-term use.

(3) In the existing eye spray on the market, if the phospholipid is added, the eye spray does not contain hyaluronic acid, because the solution system is unstable and precipitates can be generated after the hyaluronic acid is added into the phospholipid solution according to actual operation; the invention solves the problem of adding hyaluronic acid into a liposome solution, solves the problem by adding nano cross-linked hyaluronic acid into the liposome solution instead of micromolecular hyaluronic acid (although the micromolecular hyaluronic acid can also be absorbed by skin, the micromolecular hyaluronic acid can be degraded quickly and loses the moisturizing effect), and simultaneously prepares the surfactant, so that the whole liposome solution containing hyaluronic acid has good stability and quality controllability, and the storage time is more than 24 months.

(4) The preparation process of the eye spray can wrap most of the fat-soluble eye nutrient, and release the wrapped nutrient substances when the liposome microspheres enter target positions such as eyeballs, thereby achieving the effects of improving the eye comfort and nourishing the eyeballs.

Drawings

FIG. 1 is a graph of the particle size distribution of the liposome solution of example 1;

FIG. 2 is a zeta potential diagram of the liposome solution of example 1;

FIG. 3 is a graph of the particle size distribution of the liposome solution of example 2;

FIG. 4 is a zeta potential diagram of the liposome solution of example 2;

FIG. 5 is a graph of the particle size distribution of the liposome solution of example 3;

FIG. 6 is a zeta potential diagram of the liposome solution of example 3;

FIG. 7 is a graph of the particle size distribution of the liposome solution of example 4;

FIG. 8 is a zeta potential map of the liposome solution of example 4;

FIG. 9 is a graph of the particle size distribution of the liposome solution of example 5;

FIG. 10 is a zeta potential map of the liposome solution of example 5;

FIG. 11 is a graph of the particle size distribution of the liposome solution of example 6;

FIG. 12 is a zeta potential map of the liposome solution of example 6;

FIG. 13 is a graph of the particle size distribution of the liposome solution of example 7;

FIG. 14 is a zeta potential map of the liposome solution of example 7;

FIG. 15 is a graph of the particle size distribution of the liposome solution of example 8;

FIG. 16 is a zeta potential map of the liposome solution of example 8.

Detailed Description

Example 1 Liposome solution containing hyaluronic acid

The liposome solution containing hyaluronic acid of the embodiment is microemulsion, and consists of the following components in percentage by weight: 1% of phospholipid, 0.2% of cholesterol, 0.1% of nano cross-linked hyaluronic acid or sodium salt thereof, 800.20% of Tween and the balance of water.

the medical surfactant of the embodiment uses tween 80, and the medical surfactant can also be one of span and poloxamer.

The phospholipid is soybean lecithin.

the nano cross-linked hyaluronic acid or the sodium salt raw material thereof is cross-linked sodium hyaluronate gel prepared by a method of Chinese patent literature with the invention name of nano cross-linked sodium hyaluronate gel, preparation method and application thereof, which is filed in 2016 by the applicant, and the average particle size of the cross-linked sodium hyaluronate gel is 100nm to 250 nm.

The method for preparing the hyaluronic acid-containing liposome solution of this example includes the following steps:

dissolving weighed phospholipid, cholesterol and tween 80 in ethanol, shaking uniformly to obtain a lipid solution, distilling under reduced pressure to remove ethanol to obtain a lipid membrane, wherein the temperature is not more than 45 ℃ during distillation under reduced pressure.

② evenly mixing the nano cross-linked sodium hyaluronate gel with water as a dispersant to obtain hydration liquid.

mixing the lipid membrane obtained in the step one with the hydration liquid obtained in the step two to obtain a mixed solution; preparing the mixed solution into microemulsion by adopting a film-ultrasonic dispersion method (or a high-pressure homogenizer), and obtaining the liposome microemulsion solution containing hyaluronic acid.

the microemulsion solution was subjected to particle size detection and potentiometric detection using a Mastersizer model 2000 particle size analyzer from Malvern, uk.

0.1g of the prepared liposome microemulsion solution is sent into a particle size analyzer for detection, the measured particle size distribution is shown in figure 1, and the average particle size of each component in the prepared liposome microemulsion solution is 102.0 nm.

The osmolarity of the liposome solution was measured using an OM815 model freezing point osmolarity detector.

(examples 2 to 8, hyaluronic acid-containing liposome solution)

The compositions of the hyaluronic acid-containing liposome solutions of examples 2 to 8 are shown in table 1.

TABLE 1 composition of hyaluronic acid-containing liposome solution (wt%)

The mean particle diameter, zeta potential and osmotic pressure of the liposome solutions of examples 1 to 8 are shown in Table 2.

The average particle diameter, zeta potential and osmotic pressure were measured by the same apparatus as in example 1.

TABLE 2

from the detection result, the zeta potential absolute value of the liposome solution is high, the system stability is high, and the storage time can reach more than 24 months.

Example 9 preparation of hyaluronic acid-containing liposome solution

The preparation method of this example prepared the hyaluronic acid-containing liposome solution of example 1, including the following steps:

dissolving phospholipid, cholesterol and tween 80 which are weighed according to the weight percentage in ethanol, and shaking up to obtain a lipid solution.

mixing the lipid solution obtained in the step one with the hydration solution obtained in the step two, and stirring uniformly to obtain microemulsion; distilling the microemulsion at 45 deg.C under reduced pressure to remove ethanol; and (3) passing the microemulsion after removing the alcohol through a high-pressure homogenizer to obtain the liposome microemulsion solution containing the hyaluronic acid.

(example 10, eye spray)

The eye spray consists of a liposome solution containing hyaluronic acid and an eye nutrient, and specifically consists of the following components in percentage by weight: 0.5 to 2.1 percent of phospholipid, 0.1 to 0.6 percent of cholesterol, 0.05 to 0.5 percent of nano cross-linked hyaluronic acid or sodium salt thereof, 0.1 to 0.6 percent of medicinal surfactant, 0.001 to 1.05 percent of eye nutrient and the balance of water.

The eye nutritional agent is one or more of taurine, hypromellose, Va, Ve, Vc, lutein, zeaxanthin, beta-carotene, anthocyanin, DHA, and vitamin B6.

The eye spray of the embodiment consists of the liposome solution and the eye nutrient of the embodiment 1, and specifically consists of the following components in percentage by weight: phospholipid 1%, cholesterol 0.2%, nano cross-linked hyaluronic acid or sodium salt 0.1%, tween 800.20%, Va 0.025%, Ve0.002%, and water in balance.

The preparation method of the eye spray of the embodiment comprises the following steps:

dissolving weighed phospholipid, cholesterol, Va, Ve and tween 80 in ethanol, stirring at a high speed, uniformly mixing, coating the eye nutrient Va and Ve with liposome to obtain a lipid solution, distilling under reduced pressure to remove ethanol to obtain a lipid membrane, wherein the temperature is not more than 45 ℃ during distillation under reduced pressure.

Mixing the lipid membrane obtained in the step one with the hydration liquid obtained in the step two, and wrapping the eye nutrient to obtain a mixed solution; preparing the mixed solution into microemulsion by adopting a film-ultrasonic dispersion method (or a high-pressure homogenizer), and obtaining the liposome microemulsion solution containing hyaluronic acid.

(examples 11 to 19, eye spray)

the composition of the eye sprays of examples 11 to 19 is shown in table 3.

TABLE 3 composition of eye spray (wt%)

The balance in table 3 is water.

When the eye spray is prepared, the eye nutrient is added into the corresponding liposome solution and mixed evenly.

The preparation process of the eye spray can wrap most of the fat-soluble eye nutrient, and release the wrapped nutrient substances when the liposome microspheres enter target positions such as eyeballs, thereby achieving the effects of improving the eye comfort and nourishing the eyeballs.

The eye spray disclosed by the invention has the effect of treating dry eye, wherein Va and Ve added into the eye spray can also increase the elasticity of eye skin; the added cross-linked hyaluronic acid can more effectively smooth fine lines, prevent wrinkles, delay skin aging around eyes, and tighten eye bags, so that the eye skin becomes soft, tender and elastic after long-term use.

Claims

1. A liposomal microemulsion, characterized by: the composition comprises the following components in percentage by weight: 0.5 to 1.5 percent of phospholipid, 0.1 to 0.6 percent of cholesterol, 0.05 to 0.6 percent of nano cross-linked hyaluronic acid or sodium salt thereof, 0.1 to 0.6 percent of medicinal surfactant and the balance of water.

2. The liposomal microemulsion of claim 1, wherein: the particle size of the nano cross-linked hyaluronic acid or the sodium salt thereof is 50 nm-250 nm.

3. The liposomal microemulsion of claim 1, wherein: the medicinal surfactant is one of tween, span and poloxamer.

4. A method of preparing the liposomal microemulsion of claim 1, comprising the steps of:

dissolving weighed phospholipid and cholesterol in ethanol to obtain a lipid solution, and heating and evaporating to remove ethanol to obtain a lipid membrane;

Mixing the nano cross-linked sodium hyaluronate gel with water as a dispersing agent to obtain a hydration solution;

mixing the lipid membrane with the hydration liquid to obtain a mixed solution; wherein the pharmaceutically acceptable surfactant is added to the lipid solution or the hydration solution before the lipid film is mixed with the hydration solution;

preparing the mixed solution into microemulsion to obtain liposome solution containing hyaluronic acid.

5. A method of preparing the liposomal microemulsion of claim 1, comprising the steps of:

dissolving weighed phospholipid and cholesterol in ethanol to obtain a lipid solution;

6. Use of the liposomal microemulsion of claim 1 in the preparation of an ophthalmic formulation for treating or relieving dry eye, the liposomal solution being an active ingredient in the ophthalmic formulation and a carrier for an ocular nutritional agent.

7. use of a liposomal microemulsion according to claim 6 for the preparation of an ophthalmic preparation for the treatment or alleviation of dry eye, characterized in that: the ophthalmic preparation is an eye spray.

8. The eye spray according to claim 7, which is characterized by consisting of the following components in percentage by weight: 0.5 to 2.1 percent of phospholipid, 0.1 to 0.6 percent of cholesterol, 0.05 to 0.6 percent of nano cross-linked hyaluronic acid or sodium salt thereof, 0.1 to 0.6 percent of medicinal surfactant, 0.001 to 1.05 percent of eye nutrient and the balance of water.

9. The eye spray according to any one of claims 5 to 7, characterized in that: the eye nutritional agent is one or more of taurine, hypromellose, Va, Ve, Vc, lutein, zeaxanthin, beta-carotene, anthocyanin, DHA, and vitamin B6.

10. A method of preparing the eye spray of claim 7, comprising the steps of:

dissolving weighed phospholipid, cholesterol and eye nutrient in ethanol, uniformly mixing, stirring at a high speed, wrapping the eye nutrient by liposome, and heating and evaporating to remove ethanol to obtain a lipid membrane;

11. a method of preparing the eye spray of claim 7, comprising the steps of:

Dissolving weighed phospholipid, cholesterol and eye nutrient in ethanol, uniformly mixing, stirring at high speed, and coating the eye nutrient with liposome to obtain lipid solution;