Biodegradable polymer dispersion and its manufacturing method

Abstract

The present invention relates to a biodegradable polymer dispersion containing a lactic acid series polymer, a block copolymer containing a lactic acid series polymer and polyethylene glycol, and high molecular weight hyaluronic acid, and a method for producing the same, which not only allows for effective loading of functional active ingredients, but also exhibits excellent physiological activity when penetrated into the skin, has significantly improved dispersion stability, and can provide excellent delivery effects in tissues.

Description

[Technical Field]

[0001] The present invention relates to a biodegradable polymer dispersion and a method for producing the same. [Background technology]

[0002] Skin consists of the epidermis, dermis, and subcutaneous tissue. The dermis is the layer between the epidermis and subcutaneous tissue, and is the part most directly related to skin aging, containing blood vessels, collagen, elastin fibers, pores, arrector pili muscles, sebaceous glands, sweat glands, various sensory nerves, fibroblasts, and macrophages, all of which are not found in the epidermis. This is because the dermis layer is made up of more than 80% collagen, which is sensitive to UV exposure, making it susceptible to aging caused by external stimuli.

[0003] In order for an active ingredient (active ingredient) to improve skin condition through percutaneous absorption, it must penetrate the skin barrier layer present in the epidermis. However, most active ingredients cannot penetrate the barrier layer and cannot be delivered to the deeper tissues of the skin. Therefore, methods to enhance percutaneous absorption of active ingredients have been considered, such as using chemical absorption enhancers, physically forming micropores in the barrier layer, or delivering active ingredients by iontophoresis.

[0004] On the other hand, pharmaceutical approaches to enhance the penetration and permeation of active ingredients that are difficult to absorb through the skin have been extensively studied, and in particular, research has focused on the development of drug delivery systems such as colloids and nanoparticles or formulation approaches.

[0005] Nanoparticles such as liposomes, cationic polymers, quantum dots, magnetic particles, and gold nanoparticles have been studied as drug delivery systems for active ingredients, and these delivery systems are designed to promote intracellular absorption. Liposomes have the advantage of being easily designed to have targeting capabilities as physical magnetic assemblies, but they have poor colloidal stability and rely primarily on the delivery of active ingredients via diffusion through the skin barrier layer. Furthermore, cationic polymers, quantum dots, gold nanoparticles, and the like have the disadvantages of some cytotoxicity, making effective delivery of active ingredients difficult, and the delivery systems are not biodegradable.

[0006] To solve these problems, research has recently been conducted into systems that deliver or control the release of active ingredients by encapsulating the active ingredient in a carrier using a biodegradable polymer. However, this method is mainly suitable for encapsulating active ingredients with low molecular weights, and when encapsulating active ingredients with high molecular weights, it is difficult to control the release of the active ingredient and there is a problem of excessive release at the initial stage.

[0007] Therefore, there is a need for the development of an efficient dosage form that can suppress excessive initial release of an active ingredient, release the active ingredient continuously, and adjust the release rate of the active ingredient as desired. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Korean Patent Publication No. 10-2019-0095088 (August 19, 2019) Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention aims to provide a biodegradable polymer dispersion that can effectively deliver active ingredients into cells and a method for producing the same. Specifically, the present invention aims to provide a biodegradable polymer dispersion that penetrates the skin, exhibits physiological activity by itself, and is effective in improving skin conditions, and a method for producing the same.

[0010] Another object of the present invention is to provide a pharmaceutical composition containing a biodegradable polymer dispersion that contains an active ingredient inside particles containing a lactic acid-based polymer and can effectively deliver the active ingredient to the deep layers of the skin. [Means for solving the problem]

[0011] In order to solve the above-mentioned technical problems, the present invention can provide a biodegradable polymer dispersion containing a lactic acid series polymer, a block copolymer containing a lactic acid series polymer and polyethylene glycol, and high-molecular-weight hyaluronic acid having a weight-average molecular weight of 500,000 g / mol or more.

[0012] In one embodiment of the present invention, the weight ratio of the hyaluronic acid to the lactic acid series polymer may be 1.5:1 to 5:1.

[0013] In one embodiment of the present invention, the hyaluronic acid may further include one or more of a hyaluronic acid oligomer having a weight-average molecular weight of less than 6,000 g / mol and an ammonium-substituted hyaluronic acid.

[0014] In one embodiment of the present invention, the hyaluronic acid oligomer can be contained in an amount of 5 to 20% by weight based on the total weight of the hyaluronic acid.

[0015] In one embodiment of the present invention, the biodegradable polymer dispersion may have an average particle size of 0.01 to 30 μm.

[0016] In one embodiment of the present invention, the lactic acid series polymer may have a weight average molecular weight of 10,000 to 1,000,000 g / mol.

[0017] In one embodiment of the present invention, the block copolymer containing a lactic acid series polymer and polyethylene glycol may have a weight average molecular weight of 2,000 to 60,000 g / mol.

[0018] In one embodiment of the present invention, the weight ratio of the lactic acid series polymer to the block copolymer containing a lactic acid series polymer and polyethylene glycol may be 10:1 to 1:1.

[0019] In one embodiment of the present invention, the block copolymer containing a lactic acid series polymer and polyethylene glycol may be located on the surface of a particle containing the lactic acid series polymer.

[0020] The present invention can also provide a pharmaceutical composition containing the above-mentioned biodegradable polymer dispersion.

[0021] The present invention can also provide a method for producing a biodegradable polymer dispersion, comprising the steps of: dissolving a block copolymer containing a lactic acid series polymer and polyethylene glycol, and the lactic acid series polymer in a first organic solvent to produce a biodegradable polymer solution; mixing high-molecular-weight hyaluronic acid having a weight-average molecular weight of 500,000 g / mol or more with water to produce a hyaluronic acid aqueous solution; mixing the hyaluronic acid aqueous solution with the biodegradable polymer solution to produce a first dispersion; and removing the first organic solvent from the first dispersion to obtain a first emulsion.

[0022] In one embodiment of the present invention, the organic solvent may be one or a mixture of two or more selected from the group consisting of acetone, ethanol, methylene chloride, chloroform, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, and N,N-dimethylacetamide.

[0023] In one embodiment of the present invention, the hyaluronic acid aqueous solution may further contain a mixture of high molecular weight hyaluronic acid having a weight-average molecular weight of 500,000 g / mol or more, and one or more of hyaluronic acid oligomers and ammonium-substituted hyaluronic acid having a weight-average molecular weight of less than 6,000 g / mol.

[0024] In one embodiment of the present invention, the method may further include the steps of redispersing the first emulsion in a second organic solvent to prepare a second dispersion, and removing the second organic solvent from the second dispersion.

[0025] In one embodiment of the present invention, the biodegradable polymer dispersion may have a ratio of nanoparticles having an average particle size of 100 to 1000 nm of 20% or more. [Effects of the Invention]

[0026] The biodegradable polymer dispersion of the present invention contains lactic acid series polymer particles dispersed on the surface of an aqueous phase containing a hyaluronic acid mixture, in which a block copolymer (PEG-PLA) containing a lactic acid series polymer and polyethylene glycol is located. This has the advantages of having excellent dispersion stability, allowing the particle size of the lactic acid series polymer to be controlled, and allowing the dispersion to penetrate the skin by itself and having physiological activity.

[0027] Furthermore, the biodegradable polymer dispersion according to the present invention contains an active ingredient and can effectively deliver the active ingredient through the skin or other barriers. Specifically, the active ingredient can contain both hydrophilic and hydrophobic active ingredients, which has the advantage of achieving excellent physiological activity and therapeutic or ameliorative effects. Furthermore, by further including the active ingredient inside the lactic acid-based polymer particles of the biodegradable polymer dispersion, the sustained release of the active ingredient can be controlled. [Brief explanation of the drawings]

[0028] [Figure 1]1 is a graph showing the expression levels of dermal fibroblast precursor Lrig1 and papillary dermal fibroblast precursor Blimp1 when adipose-derived stem cells are treated with the biodegradable polymer dispersion according to Example 1 of the present invention and the polymers according to Comparative Examples 1 and 2. [Figure 2] 1 is a graph showing the expression levels of reticular dermis / subcutaneous fibroblast precursor Dlk1 and papillary dermal fibroblast FSP1 when adipose-derived stem cells are treated with a biodegradable polymer dispersion according to Example 1 of the present invention and the polymers according to Comparative Examples 1 and 2. [Figure 3] 1 is a graph showing mRNA expression levels of growth factors bFGF, VEGF, and HGF when adipose-derived stem cells are treated with the biodegradable polymer dispersion according to Example 1 of the present invention and the polymers according to Comparative Examples 1 and 2. [Figure 4] 1 is a graph showing the mRNA expression levels of growth factors TGF-β1, TGF-β2, and TGF-β3 when adipose-derived stem cells are treated with the biodegradable polymer dispersion according to Example 1 of the present invention and the polymers according to Comparative Examples 1 and 2. [Figure 5] 1 is a graph showing the change in moisture content (Δskin moisture) measured after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice in relation to Experimental Example 3 of the present invention. [Figure 6] 1 is a graph showing elasticity values ​​measured after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice in relation to Experimental Example 3 of the present invention. [Figure 7] In relation to Experimental Example 3 of the present invention, this figure shows microscopic images confirming changes in the basement membrane after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice. [Figure 8] 1 is a graph showing changes in the basement membrane after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice, in relation to Experimental Example 3 of the present invention. [Figure 9]In relation to Experimental Example 3 of the present invention, this figure shows microscopic images confirming changes in collagen fibers after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice. [Figure 10] 1 is a graph showing changes in collagen fibers after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice, in relation to Experimental Example 3 of the present invention. [Figure 11] In relation to Experimental Example 3 of the present invention, this figure shows microscopic images confirming changes in elastin fibers after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice. [Figure 12] 1 is a graph showing changes in elastin fibers after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice, in relation to Experimental Example 3 of the present invention. [Figure 13] In relation to Experimental Example 3 of the present invention, this figure shows microscopic images confirming changes in newborn / mature fibers 8 weeks after injection of the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice. [Figure 14] 1 is a graph showing changes in neo- and mature fibers after 8 weeks of injecting the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice, in relation to Experimental Example 3 of the present invention. [Figure 15] 1 is a graph showing changes in the expression of Tropoelastin and EBP factors after 8 weeks from the injection of the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice, in relation to Experimental Example 3 of the present invention. [Figure 16] 1 is a graph showing the expression levels of dermal fibroblast precursor Lrig1 and papillary dermal fibroblast precursor Blimp1 after 8 weeks from the injection of the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice in relation to Experimental Example 3 of the present invention. [Figure 17]1 is a graph showing the expression levels of reticular dermis / subcutaneous fibroblast precursor Dlk1 and papillary dermal fibroblast FSP1 after 8 weeks from injection of the biodegradable polymer dispersion of Example 1 and the polymers of Comparative Examples 1 and 2 into B6 mice in relation to Experimental Example 3 of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0029] The biodegradable polymer dispersion according to the present invention, its production method, and pharmaceutical compositions containing the same will be described in detail below.

[0030] Unless otherwise defined herein, all technical and scientific terms have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

[0031] In the present invention, the terms used in the description are merely for the purpose of effectively describing specific embodiments and are not intended to limit the present invention.

[0032] Furthermore, when describing components of the present invention, terms such as first, second, A, B, (a), (b), etc. may be used. These terms are used to distinguish the components from other components, and do not limit the essence, order, or procedure of the components.

[0033] Furthermore, the units of additives not specifically mentioned in this specification may be % by weight.

[0034] Also, as used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0035] Also, in this specification, the term "comprising" is an open-ended term having the same meaning as expressions such as "comprising," "containing," "having," or "characterized by," and does not exclude further unrecited elements, materials, or steps.

[0036] In the following description, descriptions of known effects and configurations that may obscure the gist of the present invention will be omitted.

[0037] The term "lactic acid series polymer particles" used in the present invention has the same meaning as "particles containing a lactic acid series polymer."

[0038] The term "PEG-PLA block copolymer" used in the present invention is used in a more specific sense of "a block copolymer containing a lactic acid-based polymer and polyethylene glycol."

[0039] The term "dispersion" used in the present invention means "a dispersion formed in an aqueous phase" and is used in a sense that includes "biodegradable polymer dispersions."

[0040] The term "skin regeneration" as used herein means reducing, weakening, delaying, or reversing the signs of skin aging or skin damage. It can also mean improving the cosmetic appearance or cosmetic enhancement of skin. Specific examples include increasing skin brightness, reducing pore size, reducing wrinkles, improving uneven skin tone due to freckles, age spots, etc., and alleviating sagging skin due to tissue loss, etc.

[0041] The present invention can provide a biodegradable polymer dispersion containing a lactic acid series polymer, a block copolymer containing a lactic acid series polymer and polyethylene glycol, and high molecular weight hyaluronic acid having a weight average molecular weight of 500,000 g / mol or more.

[0042] A lactic acid polymer refers to a polymer containing lactic acid as a structural unit, and the lactic acid may be L-lactic acid, D-lactic acid, or a combination thereof. The lactic acid unit may be contained in an amount of 50 mol% or more, specifically 60 mol% or more, and more specifically 70 mol% or more, based on 100 mol% of all monomer components constituting the lactic acid polymer, but is not limited thereto.

[0043] The lactic acid polymer may include polylactic acid (Poly(lactic acid), PLA), polylactic-co-glycolic acid copolymer (Poly(lactic acid), PLGA), etc., and specifically may be polylactic acid (Poly(lactic acid), PLA).

[0044] The lactic acid series polymer may have a weight average molecular weight of 10,000 to 1,000,000 g / mol, specifically 13,000 to 500,000 g / mol, and more specifically 15,000 to 250,000 g / mol.

[0045] The block copolymer containing a lactic acid series polymer and polyethylene glycol may refer to a polymer containing a lactic acid series polymer and a polyethylene glycol unit. Specifically, it may be a double block copolymer of a lactic acid series polymer and polyethylene glycol (PEG-PLA block copolymer). Here, the lactic acid series polymer may be hydrophobic, the polyethylene glycol may be hydrophilic, and the block copolymer may be amphiphilic.

[0046] In the block copolymer containing a lactic acid polymer and polyethylene glycol, the lactic acid polymer may be a polymer of a monomer selected from the group consisting of L-lactic acid, D-lactic acid, and L,D-lactic acid, and may have a weight-average molecular weight of 1,000 to 40,000 g / mol, specifically 1,500 to 30,000 g / mol. The block copolymer may have a weight-average molecular weight of 2,000 to 60,000 g / mol, and the polyethylene glycol in the block copolymer may have a weight-average molecular weight of 1,000 to 20,000 g / mol, specifically 3,000 to 15,000 g / mol, but is not limited thereto. In the block copolymer, the weight ratio of the lactic acid polymer block to the polyethylene glycol block may be 95:5 to 50:50, specifically 90:10 to 70:30, more specifically 90:10 to 80:20.

[0047] The lactic acid polymer and the block copolymer containing the lactic acid polymer and polyethylene glycol may be contained in a weight ratio of 10:1 to 1:1. By containing both the lactic acid polymer and the block copolymer in the above weight range, the particle size controllability and dispersion stability of the lactic acid polymer particles of the present invention can be improved, and the miscibility with the hyaluronic acid mixture described below can be improved, which is more preferable. In addition, by adjusting the weight ratio of the lactic acid polymer and the block copolymer, the size and shape of the dispersion formed in the aqueous phase can be adjusted.

[0048] More specifically, when the weight ratio of the lactic acid series polymer to the block copolymer is 6:1 to 2:1, the dispersion may be prepared into fine particles having a size of 0.01 to 4 μm, 0.1 to 1 μm, or 150 to 500 nm, and the interior of the fine particles may have a spherical shape with the lactic acid series polymer densely packed inside.

[0049] The dispersion formed in an aqueous phase containing the lactic acid series polymer and the block copolymer containing the lactic acid series polymer and polyethylene glycol may be in the form of spherical or ellipsoidal particles, and the particles may have an average particle size of 0.01 to 30 μm, specifically 0.1 to 20 μm.

[0050] In the case of a dispersion using a lactic acid series polymer alone, the average particle size may be micronized depending on the manufacturing process, but the polydispersity is high, resulting in poor reproducibility of the dispersion. In contrast, the biodegradable polymer dispersion of the present invention contains a block copolymer containing a lactic acid series polymer and polyethylene glycol, which has the advantages of being highly reproducible depending on the manufacturing process, being micronized without the application of high energy, and having high particle dispersity.

[0051] Hyaluronic acid may be in the form of a mixture that contains not only high molecular weight hyaluronic acid with a weight-average molecular weight of 500,000 g / mol or more, but also one or more of hyaluronic acid oligomer and ammonium-substituted hyaluronic acid.Specifically, it may be a mixture of high molecular weight hyaluronic acid and hyaluronic acid oligomer, a mixture of high molecular weight hyaluronic acid and ammonium-substituted hyaluronic acid, or a mixture of the above three kinds of hyaluronic acid.

[0052] In the hyaluronic acid mixture, the high molecular weight hyaluronic acid may be a high molecular weight hyaluronic acid having a weight average molecular weight of 500,000 g / mol to 3,000,000 g / mol, specifically 1,200,000 g / mol to 2,200,000 g / mol.The high molecular weight hyaluronic acid may be contained in an amount of 50 to 90 wt%, specifically 65 to 85 wt%, based on the total weight of the hyaluronic acid mixture, but is not limited thereto.

[0053] In the hyaluronic acid mixture, the ammonium-substituted hyaluronic acid may refer to a hyaluronic acid in which some or all of the hydrogen atoms of the hydroxyl groups of the hyaluronic acid are substituted with a group having a quaternary ammonium cation group, and may have a weight-average molecular weight of 300,000 g / mol to 1,000,000 g / mol, specifically 500,000 g / mol or 800,000 g / mol, and may be contained in an amount of 1 to 10 wt % of the total weight of the hyaluronic acid mixture, specifically 3 to 8 wt %, but is not limited thereto.

[0054] In the hyaluronic acid mixture, the hyaluronic acid oligomer may refer to a hydrolyzed form of hyaluronic acid. It may have a weight-average molecular weight of 800 g / mol or more but less than 8,000 g / mol, specifically a low-molecular-weight hyaluronic acid of less than 6,000 g / mol, more specifically 1,000 to 5,000 g / mol or 2,000 to 4,000 g / mol. The hyaluronic acid oligomer may be contained in an amount of 5 to 50 wt%, specifically 5 to 20 wt%, or 10 to 20 wt%, based on the total weight of the hyaluronic acid mixture, and may exhibit a viscosity suitable for skin penetration.

[0055] The weight ratio of the hyaluronic acid mixture to the lactic acid polymer may be 1.5:1 to 20:1, specifically 2:1 to 10:1, and more specifically 2:1 to 8:1. When the weight ratio falls within the above range, dispersion can be performed well during the resuspension process after freeze-drying during preparation of the biodegradable polymer dispersion.

[0056] The use of a mixture of three different types of hyaluronic acid as described above can further improve the dispersion stability of the lactic acid polymer particles in the biodegradable polymer dispersion described below. In particular, the binding of the hyaluronic acid mixture to the lactic acid polymer particles can significantly improve their interaction with the skin barrier layer. Conventional lactic acid polymers are known to be highly crystalline, have low interaction with the lipid layer that constitutes the skin barrier layer, and have significantly low adsorption or permeability to the skin barrier layer. However, when the hyaluronic acid mixture is mixed with the lactic acid polymer particles and the surface of the lactic acid polymer particles is hydrated, the interaction of the lactic acid polymer particles with the skin barrier layer is significantly improved, allowing them to strongly adsorb to or permeate the skin barrier layer. This is preferable because it provides excellent skin penetration when the biodegradable polymer dispersion is applied to the skin, further improving physiological activities such as improving skin cell elasticity, inhibiting aging, and promoting skin cell growth.

[0057] A preferred embodiment of the biodegradable polymer dispersion of the present invention comprises lactic acid-based polymer particles dispersed in a continuous phase containing a hyaluronic acid mixture, with a polylactic acid-polyethylene glycol block copolymer (PLA-PEG diblock copolymer) positioned on the surface of the lactic acid-based polymer particles. More specifically, the hydrophobic polylactic acid portion of the block copolymer may be positioned facing the lactic acid-based polymer particles, and the hydrophilic polyethylene glycol portion may be positioned facing the continuous phase. A hyaluronic acid mixture containing three different types of hyaluronic acid forming the continuous phase allows the hydrophobic portion of the block copolymer to be positioned at a higher density on the particle surface, and the hydrophilic polyethylene glycol portion provides excellent dispersion stability, allowing a large amount of lactic acid-based polymer particles, which are the dispersed phase, to be accommodated within the continuous phase, which is more preferred. This allows the biodegradable polymer dispersion of the present invention to contain active ingredients independently in the continuous and dispersed phases, more preferably.

[0058] The method for producing a biodegradable polymer dispersion according to the present invention will be described in detail below.

[0059] The method for producing a biodegradable polymer dispersion according to the present invention includes the steps of dissolving a block copolymer containing a lactic acid series polymer and polyethylene glycol, and the lactic acid series polymer in a first organic solvent to produce a biodegradable polymer solution, mixing hyaluronic acid and water to produce a hyaluronic acid aqueous solution, mixing the biodegradable polymer solution with the hyaluronic acid aqueous solution to produce a first dispersion, and removing the first organic solvent from the first dispersion to obtain a first emulsion.

[0060] In addition, by further including the steps of redispersing the first emulsion in a second organic solvent to prepare a second dispersion, and removing the second organic solvent from the second dispersion, the effect of further improving dispersion stability during resuspension can be obtained.

[0061] The step of preparing the biodegradable polymer solution may be carried out by dissolving the lactic acid-based polymer and the block copolymer in an organic solvent. Specifically, the organic solvent may be one or a mixture of two or more selected from the group consisting of acetone, ethanol, methylene chloride, chloroform, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, and N,N-dimethylacetamie, and preferably acetone.

[0062] The organic solvent may be contained in an amount of 70% by weight to 95% by weight based on the total weight of the biodegradable polymer solution, but is not limited thereto.

[0063] The hyaluronic acid aqueous solution can be prepared by mixing the above-mentioned high molecular weight hyaluronic acid, two kinds of hyaluronic acid, high molecular weight hyaluronic acid and low molecular weight hyaluronic acid, or three kinds of hyaluronic acid, two kinds of hyaluronic acid and ammonium-substituted hyaluronic acid, with water, so that the water content of the total aqueous solution is 95% by weight to 99.9% by weight, but is not limited thereto.Here, the mixing ratio of the two or three kinds of hyaluronic acid is as described above.

[0064] The step of preparing the first dispersion may involve mixing a biodegradable polymer solution and an aqueous hyaluronic acid solution in a volume ratio of 1:1 to 1:3. Here, the first dispersion may have an aqueous solution that forms a continuous phase, an organic solvent that is extracted onto the continuous phase, and a dispersed phase of the phase-separated polymer. Specifically, the dispersed phase may have a block copolymer containing a lactic acid polymer and polyethylene glycol located on the surface of dispersed lactic acid polymer particles.

[0065] The step of removing the organic solvent is not limited, but specifically, evaporation may be used, and the evaporation may be performed at 20 to 130°C for 1 to 48 hours, but is not limited thereto, and the evaporation conditions can be adjusted depending on the type of solvent used. Furthermore, the evaporation may be performed under reduced pressure.

[0066] After removing the first organic solvent to obtain the first emulsion, the first emulsion can be redispersed in a second organic solvent to prepare a second dispersion. In a preferred embodiment of the present invention, the method further includes a step of freeze-drying the obtained first emulsion to obtain the first dispersion in a solid state, which can then be redispersed in a second organic solvent to prepare a second dispersion. In this case, the resuspension can be performed more stably.

[0067] The biodegradable polymer dispersion of the present invention produced through the above steps may have the lactic acid series polymer particles dispersed on a continuous phase containing the hyaluronic acid mixture, and the lactic acid series polymer particles may have a block copolymer containing a lactic acid series polymer and polyethylene glycol located on the surface.

[0068] The biodegradable polymer dispersion according to one embodiment of the present invention has excellent physiologically active properties, biocompatibility, and high loading properties that allow it to contain a large amount of active ingredients, and is therefore applicable to various fields. Specifically, it can be used in cosmetic compositions, topical skin compositions, pharmaceutical compositions, etc., that contain the biodegradable polymer dispersion.

[0069] For example, a pharmaceutical composition for skin regeneration containing the biodegradable polymer dispersion can be provided. Specifically, the present invention can induce collagen synthesis to provide local skin regeneration effects in soft tissues. More specifically, when injected around a wound in any part of the body, such as the face, neck, chest, buttocks, arms, armpits, hands, legs, or feet, it can promote wound healing, inhibit scar formation, or provide scar treatment effects at the wound site.

[0070] As used herein, "soft tissue" refers to tissue that is not bone but connects, supports, or surrounds other structures and organs of the body. Specifically, injectable soft tissue can include muscles, ligaments, fascia, skin, dermis, fibrous tissue, fat, synovial membrane, muscle, nerves, and blood vessels.

[0071] As an example, a cosmetic composition for wrinkle reduction containing the biodegradable polymer dispersion can be provided. A composition containing the biodegradable polymer dispersion according to the present invention can activate fibroblasts or fibroblast precursors to induce collagen production. Fibroblasts stimulate the production of structural proteins such as collagen and elastin, and the collagen produced by fibroblasts can form fibrous connective tissue that maintains skin elasticity. By injecting the composition according to the present invention into the site of skin injury, it is possible to achieve scar and wrinkle reduction effects.

[0072] For example, a pharmaceutical composition for skin regeneration or wound treatment containing the biodegradable polymer dispersion can be provided. Here, the pharmaceutical composition can contain a pharmaceutically acceptable carrier, if necessary, and can be formulated into a transdermal dosage form such as a solution, suspension, emulsion, lotion, or ointment according to a conventional method.

[0073] The pharmaceutically acceptable carrier may include aqueous diluents or solvents such as phosphate buffered saline, purified water, or sterile water, and may also include non-aqueous diluents or solvents such as propylene glycol or olive oil.

[0074] The pharmaceutical composition may vary depending on the condition and severity of the patient, the form of the drug, and the route and duration of administration, and can be appropriately selected by a person skilled in the art.

[0075] Furthermore, the following three modes are possible: a first mode in which a hydrophilic active ingredient is contained in a hyaluronic acid mixture, which is a hydrophilic continuous phase; a second mode in which a hydrophobic active ingredient is contained inside a hydrophobic lactic acid-based polymer; and a third mode in which an active ingredient is contained inside both the hyaluronic acid mixture, which is a hydrophilic continuous phase, and the hydrophobic lactic acid-based polymer.

[0076] The active ingredient can be any known active ingredient that is physiologically active on the skin or other tissues, and may further contain conventional additives that are not harmful to the human body, such as additional fragrances, vitamins, stabilizers, and antioxidants, within the range that does not impair the physical properties of the biodegradable polymer dispersion of the present invention.

[0077] The biodegradable polymer dispersion of the present invention or a composition containing the biodegradable polymer dispersion can be applied in the form of known internal preparations and external preparations, and preferably, it may be applied as an external preparation such as a cream, ointment, lotion, or gel.

[0078] The present invention can also provide a method for regenerating skin or treating skin scars, which comprises the step of administering a pharmaceutical composition containing the biodegradable polymer dispersion described above.

[0079] The biodegradable polymer dispersion or a composition containing the biodegradable polymer dispersion can be directly applied to the skin, and when the applied area is subjected to plasma treatment, the penetration of the biodegradable polymer dispersion into the skin increases, and as absorption into the skin is promoted, the physiological activity properties can be accelerated, allowing for effective skin improvement in a short period of time, which can be useful for treatment through skin regeneration and is applicable to skin improvement.

[0080] In addition, when the biodegradable polymer dispersion contains an additional active ingredient, the plasma treatment can promote diffusion of the active ingredient into the deeper layers of the skin, thereby further maximizing the skin improvement or treatment effect, which is preferable.

[0081] The biodegradable polymer dispersion or a composition containing the same according to an embodiment of the present invention having the above-described properties can be used as a preferred skin care system when combined with a plasma treatment device.

[0082] The present invention will be described in more detail below with reference to examples and comparative examples, but the following examples and comparative examples are merely illustrative examples for explaining the present invention in more detail, and the present invention is not limited by the following examples and comparative examples.

[0083] [Example 1] A biodegradable polymer solution was prepared by mixing 0.25 g of polylactic acid (EVONIK, RESOMER RR 202S) and 0.125 g of PEG-PLA block copolymer (EVONIK, RESOMER 100 DL mPEG5000) in 25 mL of acetone.

[0084] An aqueous solution was prepared by mixing 9.0 g of hyaluronic acid (molecular weight 1,200,000 g / mol) and 1.0 g of sodium hyaluronate (molecular weight 5,000 g / mol) hydrolyzed to hyaluronic acid oligomer with 1,000 mL of distilled water.

[0085] Next, 25 mL of the prepared polymer solution was gradually mixed with 50 mL of aqueous solution under stirring, and after the mixture was completed, acetone was evaporated under reduced pressure at room temperature to obtain an emulsion, which was then freeze-dried to obtain a dispersion, which was then re-dispersed in an organic solvent, and the organic solvent was evaporated under reduced pressure to prepare a biodegradable polymer dispersion.

[0086] [Example 2] A biodegradable polymer dispersion was produced in the same manner as in Example 1, except that an aqueous hyaluronic acid solution was produced by mixing 9.0 g of hyaluronic acid (molecular weight 1,200,000 g / mol) and 1.0 g of hydroxypropyltriammonium hyaluronic acid (molecular weight 500,000 g / mol), an ammonium-substituted hyaluronic acid, with 1,000 mL of distilled water.

[0087] [Example 3] A biodegradable polymer dispersion was produced in the same manner as in Example 1, except that 10.0 g of hyaluronic acid (molecular weight 1,200,000 g / mol) was mixed with 1,000 mL of distilled water.

[0088] [Example 4] A biodegradable polymer dispersion was prepared in the same manner as in Example 1, except that 0.25 g of polylactate-co-glycolate (manufactured by EVONIK, RESOMER RG 752 S) was used instead of polylactic acid, and 0.125 g of PEG-PLA copolymer was used.

[0089] [Comparative Example 1] A hydrogel filler sample was prepared using 1,4-butanediol diglycidyl ether (BDDE) as a crosslinker, which is a crosslinked hyaluronic acid product containing small particles.

[0090] Comparative Example 2 A biodegradable polymer solution was prepared by dissolving 2.5 g of PLA in 20 mL of ethyl acetate, and then dispersed in 100 mL of water containing 5 g of Tween 80 (polyoxyethylene sorbitan monooleate). The solvent was then evaporated to prepare a biodegradable polymer dispersion.

[0091] Comparative Example 3 A biodegradable polymer dispersion was produced in the same manner as in Example 1, except that 0.125 g of polylactic acid was used in Example 1, and an aqueous solution was produced by mixing 2.125 g of hyaluronic acid (molecular weight 1,200,000 g / mol), 0.125 g of hydroxypropyltriammonium hyaluronic acid (molecular weight 500,000 g / mol) which is an ammonium-substituted hyaluronic acid, and 0.25 g of sodium hyaluronate (molecular weight 5,000 g / mol) hydrolyzed to hyaluronic acid oligomer in 1,000 mL of distilled water.

[0092] Comparative Example 4 A biodegradable polymer dispersion was produced in the same manner as in Example 1, except that an aqueous solution was produced by mixing 2.125 g of hyaluronic acid (molecular weight 1,200,000 g / mol), 0.125 g of hydroxypropyltriammonium hyaluronic acid (molecular weight 500,000 g / mol) as ammonium-substituted hyaluronic acid, and 0.25 g of sodium hyaluronate (molecular weight 5,000 g / mol) hydrolyzed to hyaluronic acid oligomer with 1,000 mL of distilled water.

[0093] Comparative Example 5 A biodegradable polymer dispersion was produced in the same manner as in Example 1, except that 0.125 g of polylactic acid was used in Example 1, and an aqueous solution was produced by mixing 0.7 g of hyaluronic acid (molecular weight 1,200,000 g / mol), 0.04 g of hydroxypropyltriammonium hyaluronic acid (molecular weight 500,000 g / mol) as ammonium-substituted hyaluronic acid, and 0.08 g of sodium hyaluronate (molecular weight 5,000 g / mol) hydrolyzed to hyaluronic acid oligomer in 1,000 mL of distilled water.

[0094] The properties of the examples and comparative examples were evaluated as follows.

[0095] [Experimental Example 1] Preparation of biodegradable polymer dispersion and evaluation of redispersibility The biodegradable polymer dispersions prepared in Examples 1 to 4 and Comparative Examples 3 to 5 were resuspended in distilled water, and the emulsion state was observed. The particle size of the dispersed particles in the biodegradable polymer dispersion was measured, and the nanoparticle % is shown in Table 1 below. Here, nanoparticles were defined as those having an average particle size of 100 to 1000 nm.

[0096] In Examples 1 to 4, the nanoparticle portion was at least about 20%, confirming that nanoparticles were successfully formed, but in the biodegradable polymer dispersions of Comparative Examples 3 to 5, the nanoparticle portion was less than 5%, indicating that nanoparticle formation was not carried out properly.Compared to Comparative Examples 3 to 5, it can be confirmed that the examples according to the present invention showed significantly better nanoparticle formation.

[0097] [Table 1]

[0098] [Experimental Example 2] Evaluation of the efficacy of biodegradable polymer dispersions on stem cells (in vitro) Human adipose-derived mesenchymal stem cells (hAD-MSCs) were cultured in Human MSC Growth Medium (CEFOgro TM )60cm 2 The cultured hAD-MSCs were treated with 120 μl of the samples prepared in Example 1, Comparative Example 1, and Comparative Example 2, and PBS for 48 hours. RNA was extracted from the sample-treated hAD-MSCs using RNAiso Plus (Cat. 9109) reagent (Takara, Japan) according to the prescribed protocol. The extracted RNA was quantified, and then cDNA was synthesized. The cDNA was synthesized using PrimeScript TM The cDNA was synthesized using a 1st strand cDNA Synthesis Kit (Cat. 6110A) (Takara, Japan) according to the protocol. To confirm the mRNA level using the synthesized cDNA, 0.2 μM of the target gene primer and 200 ng of the cDNA mixture were incubated with SYBR TM green and quantitative gene amplification experiments were performed.

[0099] Next, the secretion levels of growth factors and dermal fibroblast precursors in adipose-derived stem cells in the biodegradable polymer dispersions of Example 1 and Comparative Examples 1 and 2 were measured, and the results are shown in FIGS.

[0100] [Experimental Example 3] Evaluation of the elasticity-improving effect of biodegradable polymer dispersion (in vivo) Five 12-month-old male C57BL6J (B6) mice were used as one collagen group, and 100 μl of each of the samples from Example 1, Comparative Examples 1 and 2 was injected into the skin at five locations using a 27G screw needle. After 8 weeks, the mice included in the collagen group were culled.

[0101] 1. Moisture and elasticity measurement Moisture and elasticity were measured at two points before the sample was treated and before the tissue was harvested after 8 weeks. After removing the hair and leaving the skin smooth, five values ​​were measured using a skin analyzer, and three similar values ​​were read and shown in Figures 5 and 6.

[0102] 2. Tissue collection and testing After removing the hair from the mice, the tissue surrounding the treated area was widely excised and visually inspected to see if any sample remained within the skin. The tissue was then cut into several equal pieces, placed in clean tubes, cooled using liquid nitrogen, and stored in an ultra-low temperature refrigerator for fixation. The fixed skin tissue was then processed into paraffin blocks using a tissue processor and tissue embedding device, and cut into 7 μm sections using a milling machine for histological examination.

[0103] (1) Changes in the basement membrane were confirmed by Periodic Acid Schiff (PAS) staining using a Periodic Acid Schiff staining kit (Cat. SSK5020, BBC biochemical). The results are shown in Figures 7 and 8. The basement membrane is located between the epidermal layer and the dermal layer, and PAS-positive signals are expressed in dark red. Example 1 showed a statistically significant increase in PAS-positive signals compared to aged mice (control), but there was no change in Comparative Examples 1 and 2.

[0104] (2) Changes in collagen fibers were confirmed by Masson trichrome (MT) staining using a Trichrome Staining Kit (Modified Masson's) (Cat. TRM-IFU, ScyTek Laboratories). The results are shown in Figures 9 and 10. After MT staining, the blue signal in the dermis layer of the skin tissue represents collagen fibers. Example 1 showed a statistically significant increase in MT-positive signals compared to aged mice (control), but there was no change in Comparative Examples 1 and 2.

[0105] (3) Using an Elastic Staining Kit (Modified Verhoff's) (Cat. ETS-1-IFU, ScyTek Laboratories), changes in elastin fibers were confirmed by Verhoff staining. The results are shown in Figures 11 and 12. After Verhoff staining, the blue signal in the dermis layer of the skin tissue represents elastic fibers. Example 1 showed a statistically significant increase in Verhoff positive signals compared to aged mice (control), but there was no change in Comparative Examples 1 and 2.

[0106] (4) Using a Herovici staining kit (Cat. HSK-IFU, ScyTek Laboratories), changes in neo- and mature collagen fibers were confirmed by Herovici staining. The results are shown in Figures 13 and 14. After Herovici staining, the blue signals in the dermis layer of the skin tissue represent neo- and mature collagen fibers. Example 1 showed a statistically significant increase in Verhoeff-positive signals compared to aged mice (control), but there was no change in Comparative Examples 1 and 2.

[0107] (5) RNA was extracted from 100 mg of tissue stored in an ultra-low temperature refrigerator using RNAiso Plus (Cat. 9109, Takara) reagent. The extracted RNA was quantified and then analyzed using PrimeScript. TMcDNA was synthesized using a 1st strand cDNA Synthesis Kit (Cat. 6110a, Takara). To confirm the mRNA level using the synthesized cDNA, 0.2 μM of target gene primer and 200 ng of cDNA were mixed with SYBR TM Quantitative gene amplification experiments were performed by mixing with IgG1 and IgG2a. Data are expressed as the mean ± standard error of the mean (SEM) of three independent experiments (*, P<0.05).

[0108] As described above, the present invention has been described using specific and limited examples and drawings, but these are provided to facilitate a more general understanding of the present invention, and the present invention is not limited to the above examples. Various modifications and variations can be made from such descriptions by those having ordinary knowledge in the field to which the present invention pertains.

[0109] Therefore, the concept of the present invention should not be limited to the above-described embodiments, and it can be said that not only the scope of the claims described below, but also all modifications that are equivalent to or equivalent to the scope of the claims fall within the scope of the concept of the present invention.

Claims

1. A biodegradable polymer dispersion comprising a lactic acid series polymer, a block copolymer containing a lactic acid series polymer and polyethylene glycol, and high-molecular-weight hyaluronic acid having a weight-average molecular weight of 500,000 g / mol or more.

2. 2. The biodegradable polymer dispersion according to claim 1, wherein the weight ratio of the hyaluronic acid to the lactic acid polymer is 1.5:1 to 5:

1.

3. The biodegradable polymer dispersion according to claim 1, wherein the hyaluronic acid further comprises one or more of a hyaluronic acid oligomer having a weight-average molecular weight of less than 6,000 g / mol and an ammonium-substituted hyaluronic acid.

4. The biodegradable polymer dispersion according to claim 3, wherein the hyaluronic acid oligomer is contained in an amount of 5 to 20% by weight based on the total weight of the hyaluronic acid.

5. 2. The biodegradable polymer dispersion according to claim 1, which is in the form of spherical particles having an average particle size of 0.01 to 30 μm.

6. 2. The biodegradable polymer dispersion according to claim 1, wherein the lactic acid series polymer has a weight average molecular weight of 10,000 to 1,000,000 g / mol.

7. 2. The biodegradable polymer dispersion according to claim 1, wherein the block copolymer containing a lactic acid series polymer and polyethylene glycol has a weight average molecular weight of 2,000 to 60,000 g / mol.

8. 2. The biodegradable polymer dispersion according to claim 1, wherein the weight ratio of the lactic acid series polymer to the block copolymer containing the lactic acid series polymer and polyethylene glycol is 10:1 to 1:

1.

9. The biodegradable polymer dispersion according to claim 1 , wherein the block copolymer containing the lactic acid series polymer and polyethylene glycol is located on the surface of a particle containing the lactic acid series polymer.

10. A pharmaceutical composition comprising the biodegradable polymer dispersion of claim 1.

11. a step of dissolving a block copolymer containing a lactic acid series polymer and polyethylene glycol, and the lactic acid series polymer in a first organic solvent to prepare a biodegradable polymer solution; A step of mixing high molecular weight hyaluronic acid having a weight average molecular weight of 500,000 g / mol or more and water to prepare a hyaluronic acid aqueous solution; mixing the biodegradable polymer solution with a hyaluronic acid aqueous solution to prepare a first dispersion; and removing the first organic solvent from the first dispersion to obtain a first emulsion.

12. The method for producing a biodegradable polymer dispersion according to claim 11, wherein the organic solvent is one or a mixture of two or more selected from the group consisting of acetone, ethanol, methylene chloride, chloroform, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, and N,N-dimethylacetamide.

13. The method for producing a biodegradable polymer dispersion according to claim 11, wherein the hyaluronic acid aqueous solution further contains at least one of a hyaluronic acid oligomer having a weight average molecular weight of less than 6,000 g / mol and an ammonium-substituted hyaluronic acid.

14. redispersing the first emulsion in a second organic solvent to produce a second dispersion; The method for producing a biodegradable polymer dispersion according to claim 11, further comprising the step of removing the second organic solvent from the second dispersion.

15. The method for producing a biodegradable polymer dispersion according to claim 14, wherein the biodegradable polymer dispersion contains nanoparticles with an average particle size of 100 to 1000 nm in an amount of 20% or more.

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