Powder preparation for tissue repair and injection composition for tissue repair comprising same

Abstract

The present invention relates to a powder preparation for tissue repair and an injection composition for tissue repair comprising the powder preparation, and more particularly, to a powder preparation for tissue repair and an injection composition for tissue repair comprising the powder preparation, which are characterized in that: the powder preparation is a powder preparation for tissue repair; the present invention relates to a powder formulation comprising a copolymer of a hydrophilic biocompatible polymer and a hydrophobic biocompatible polymer, in which when a reference solution comprising a specific concentration of the powder formulation is diluted according to a series of increased dilution multiples, the concentration of the reference solution is greater than the concentration of the reference solution. The number of polymer particles in the diluted solution is greater than or equal to the number of polymer particles in the reference solution in a specific dilution multiple interval, and then the number of polymer particles shows a decreasing behavior as the dilution multiple increases.

Description

Technical Field

[0001] The present invention relates to a powder preparation for tissue repair treatment and an injectable composition for tissue repair treatment containing the powder preparation. More specifically, the present invention relates to a powder preparation and an injectable composition for tissue repair treatment containing the powder preparation, characterized in that the powder preparation contains a copolymer of a hydrophilic biocompatible polymer and a hydrophobic biocompatible polymer. When a reference solution containing the powder preparation at a specific concentration is diluted according to a series of increasing dilution multiples, the number of polymer particles in the diluted solution is greater than or equal to the number of polymer particles in the reference solution in a specific dilution multiple range, and then as the dilution multiple increases, the number of the polymer particles shows a decreasing behavior. Background Art

[0002] In recent years, many people have been interested in "well-aging", that is, aging gracefully from a young age. It is no exaggeration to say that the current beauty market is an era of anti-aging that focuses on aging slowly, beautifully and healthily. The most common phenomenon of aging is the reduction of plumpness. In particular, if the face lacks plumpness, it will look older and haggard, so people are very interested in fillers that can provide plumpness. Therefore, the filler market is also growing rapidly every year and currently forms a market of more than 2 trillion globally.

[0003] Currently, various substances are used as filler materials. Among them, hyaluronic acid fillers account for more than 90% of the global filler market and have a half-life in the body within 1 to 3 days. Therefore, there are problems of very low persistence and very fast resorption in the body. Therefore, products that extend the resorption period by connecting hyaluronic acid and a cross-linking substance have emerged on the market. However, in the case of such cross-linked products, 1,4-butanediol diglycidylether (BDDE), which is a cross-linking substance, is a toxic carcinogen. Therefore, there are problems of losses such as an increase in process costs according to its removal process, product abandonment due to microbial contamination, and product abandonment due to the detection of residues.

[0004] Therefore, many products for tissue repair using polymers that decompose in vivo have also been developed. As filler preparations using conventional biocompatible polymers, preparations in which water-insoluble polymers are processed into micron-sized particles and then dispersed through a viscous excipient or thickener have been developed and used. For example, a preparation in which poly(lactic acid) (PLA) particles with a diameter of 20 to 50 microns are dispersed in an aqueous solution of carboxymethylcellulose (CMC), or a preparation in which poly(caprolactone) (PCL) particles with a diameter of 20 to 50 microns are dispersed in an aqueous solution of CMC and glycerin has been used. However, such preparations have operational inconveniences such as the needle being blocked by microparticles during injection, and problems such as an uneven tissue repair effect being unable to be produced due to uneven particle dispersion. Summary of the Invention

[0005] Technical Problem to be Solved

[0006] An object of the present invention is to provide a product for tissue repair and a preparation method thereof that have no risk of toxicity, can ensure the functionality, physical properties, and safety of a biomaterial suitable for tissue repair, and show excellent tissue repair effects while being easy to transport, store, and handle.

[0007] Technical Solution

[0008] One aspect of the present invention provides a powder preparation for tissue repair treatment, characterized in that the powder preparation contains a biocompatible copolymer that is a copolymer of a hydrophilic biocompatible polymer and a hydrophobic biocompatible polymer.

[0009] When a reference solution containing the powder preparation at a concentration of 7.5% by weight is diluted according to a series of dilution multiples that increase sequentially,

[0010] in the dilution multiple range of 4 to 8, the number of polymer particles per unit volume in the diluted solution is greater than or equal to the number of polymer particles per unit volume in the reference solution.

[0011] In the dilution multiple range of 16 or more, as the dilution multiple increases, the number of polymer particles per unit volume in the diluted solution decreases.

[0012] Another aspect of the present invention provides an injection composition for tissue repair treatment, the injection composition comprising: the powder preparation for tissue repair treatment of the present invention; and a pharmaceutically acceptable injection carrier.

[0013] Another aspect of the present invention provides a method for preparing an injectable composition for tissue repair treatment, the preparation method comprising: mixing the powder preparation for tissue repair treatment of the present invention and a pharmaceutically acceptable injectable carrier at room temperature.

[0014] Beneficial effects

[0015] The powder preparation for tissue repair treatment according to the present invention is non-toxic, easy to transport, store and handle, forms polymer particles that are easily dispersed and dissolved in an aqueous medium at room temperature, and after being introduced into the body, it is not phagocytosed by macrophages and induces collagen, thereby showing excellent tissue repair effects. Description of the drawings

[0016] Figure 1 Shows the results of measuring the number of particles of Preparation Examples 1 to 4 carried out in Experimental Example 2 of the present invention.

[0017] Figure 2 Shows the results of measuring the number of particles of Comparative Examples 1 to 5 carried out in Experimental Example 2 of the present invention.

[0018] Figure 3 Shows the results of the animal experiment on tissue repair effect carried out in Experimental Example 3 of the present invention. Detailed description of the invention

[0019] Hereinafter, the present invention will be described in more detail.

[0020] The powder preparation for tissue repair treatment of the present invention contains a biocompatible copolymer that is a copolymer of a hydrophilic biocompatible polymer and a hydrophobic biocompatible polymer.

[0021] The powder preparation for tissue repair treatment of the present invention is characterized in that when a reference solution containing the powder preparation at a concentration of 7.5% by weight is diluted according to a series of subsequently increasing dilution factors, (i) in the dilution factor range of 4 to 8, the number of polymer particles per unit volume in the diluted solution is greater than or equal to the number of polymer particles per unit volume in the reference solution, and (ii) in the dilution factor range above 16, as the dilution factor increases, the number of polymer particles per unit volume in the diluted solution decreases.

[0022] In a specific embodiment, the medium for the preparation and dilution of the reference solution may be an aqueous medium, for example, it may be phosphate buffer solution, water for injection, normal saline, distilled water, pure water or glucose solution.

[0023] In a specific embodiment, the series of dilution multiples that increase successively may include dilution multiples of 2, 4, 8, 16, 32, and 64, but are not limited thereto.

[0024] In this specification, the "dilution multiple" refers to the volume multiple of the dilution solution to the reference solution when diluting the reference solution. For example, a "dilution solution with a dilution multiple of 4" refers to a dilution solution prepared by adding a medium equivalent to 3 times the volume of the reference solution to the reference solution so that the final volume is 4 times the volume of the reference solution.

[0025] In this specification, the "dilution multiple range" is determined by the dilution multiple applied when diluting the reference solution. For example, the "dilution multiple range from 4 to 8" includes dilution solutions with a minimum dilution multiple of 4 and a maximum of 8, and the "dilution multiple range above 16" includes dilution solutions with a dilution multiple above 16.

[0026] In this specification, "in certain dilution multiple ranges, as the dilution multiple increases, the number of polymer particles per unit volume in the dilution solution decreases" means that the number of polymer particles per unit volume in a solution with a dilution multiple within this dilution multiple range is greater than the number of polymer particles per unit volume in a solution with a dilution multiple higher than this dilution multiple. In the present invention, in the dilution multiple range above 16, as the dilution multiple increases, the number of polymer particles per unit volume in the dilution solution decreases. Therefore, for example, the number of polymer particles per unit volume in a solution with a dilution multiple of 16 is greater than the number of polymer particles per unit volume in a solution with a dilution multiple of 32.

[0027] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may be characterized in that in the dilution multiple range from 4 to 16, the number of polymer particles per unit volume in the dilution solution is greater than or equal to the number of polymer particles per unit volume in the reference solution.

[0028] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may be characterized in that in the dilution multiple range from 4 to 32, the number of polymer particles per unit volume in the dilution solution is greater than or equal to the number of polymer particles per unit volume in the reference solution.

[0029] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may be characterized in that in the dilution multiple range above 8, as the dilution multiple increases, the number of polymer particles per unit volume in the dilution solution decreases.

[0030] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may be characterized in that in the dilution multiple range above 4, as the dilution multiple increases, the number of polymer particles per unit volume in the dilution solution decreases.

[0031] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may be characterized in that when the number of polymer particles per unit volume in the reference solution is set to 100%, at a dilution multiple of 32, the number of polymer particles per unit volume in the dilution solution is 60% or more (for example, 60% to 99%).

[0032] More specifically, the powder preparation for tissue repair treatment of the present invention may be characterized in that when the number of polymer particles per unit volume in the reference solution is set to 100%, at a dilution multiple of 32, the number of polymer particles per unit volume in the dilution solution can be 65% or more, 70% or more, 75% or more, or 80% or more.

[0033] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may be characterized in that when the number of polymer particles per unit volume in the reference solution is set to 100%, at a dilution multiple of 64, the number of polymer particles per unit volume in the dilution solution is 40% or more (for example, 40% to 59%).

[0034] More specifically, the powder preparation for tissue repair treatment of the present invention may be characterized in that when the number of polymer particles per unit volume in the reference solution is set to 100%, at a dilution multiple of 64, the number of polymer particles per unit volume in the dilution solution can be 45% or more, 50% or more, or 55% or more.

[0035] In a specific embodiment, the number of polymer particles per unit volume in the solution can be measured by flow cytometry (Flow Cytometer, FC). Flow cytometry is a method of measuring the number of particles by counting the number of events scattered into the detection area while irradiating with a laser while flowing a solution containing polymer particles at a specific flow rate for a specific time. For example, it can be performed using an APOGEE device. When using an APOGEE device, the number of particles having an average diameter in the range of 20 to 5000 nm can be measured. That is, according to a specific embodiment, the polymer particles contained in the powder preparation for tissue repair treatment of the present invention can be particles having an average diameter of 20 to 5000 nm.

[0036] In a specific embodiment, the hydrophilic biocompatible polymer may be selected from polyethylene glycol or its derivatives (e.g., alkoxy-polyethylene glycol or hydroxy-polyethylene glycol), polyvinylpyrrolidone, polyvinyl alcohol, polyacrylamide, and combinations thereof. More specifically, it may be selected from polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), and combinations thereof.

[0037] In a specific embodiment, the hydrophobic biocompatible polymer may be a polymer of monomers derived from α-hydroxy acids. More specifically, it may be selected from polylactide, polyglycolide, poly(lactide-glycolide), polytropic acid, polycaprolactone, poly(dioxanone-2-one), polyamino acids, polyorthoesters, polyanhydrides, polycarbonates, and combinations thereof. Even more specifically, it may be selected from polylactide, polyglycolide, poly(lactide-glycolide), and combinations thereof.

[0038] Specifically, the number average molecular weight (Mn1) (unit: g / mol) of the hydrophilic biocompatible polymer according to GPC may be 1000 or more, 2000 or more, 3000 or more, or 4000 or more, and may be 30000 or less, 25000 or less, 20000 or less, 19000 or less, 18000 or less, 17000 or less, 16000 or less, or 15000 or less.

[0039] Specifically, the number average molecular weight (Mn2) (unit: g / mol) of the hydrophobic biocompatible polymer according to GPC may be 500 or more, 1000 or more, 1500 or more, or 2000 or more, and may be 30000 or less, 25000 or less, 20000 or less, 19000 or less, 18000 or less, 17000 or less, 16000 or less, or 15000 or less.

[0040] Specifically, the total number average molecular weight of the biocompatible copolymer according to GPC may be 2000 or more, 5000 or more, 8000 or more, or 10000 or more, and may be 40000 or less, 35000 or less, 30000 or less, or 25000 or less.

[0041] In a specific embodiment, the ratio (Mn1 / Mn2) of the number average molecular weight (Mn1) of the hydrophilic biocompatible polymer to the number average molecular weight (Mn2) of the hydrophobic biocompatible polymer in the biocompatible copolymer may be 2.5 or less.

[0042] The number average molecular weights of the hydrophilic biocompatible polymer and the hydrophobic biocompatible polymer may be measured by, for example, Gel Permeation Chromatography (GPC).

[0043] Gel permeation chromatography (GPC) is a method in which the physical elution mechanism is based on the size of the hydrodynamic volume of the components to be analyzed. Larger components are eluted first, and smaller components are eluted later. Molecules with a large size cannot enter the pores of the porous gel, so they pass through quickly, while small molecules can enter the pores of the gel, stay there, and thus pass through slowly. The relative molecular weights are analyzed in the order of the molecules that pass through the chromatographic column faster.

[0044] More specifically, the Mn1 / Mn2 ratio in the biocompatible copolymer can be 2.5 or less, 2.3 or less, 2.1 or less, 2.0 or less, or 1.8 or less. The lower limit of the Mn1 / Mn2 ratio is not particularly limited. For example, it can be 0.5 or more, 0.6 or more, 0.7 or more, 0.8 or more, or 0.9 or more, but it is not limited thereto.

[0045] In addition to the above biocompatible copolymer, the powder preparation for tissue repair treatment of the present invention may further contain one or more additives that can be commonly used in pharmaceutical powder preparations.

[0046] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may further contain a lyophilization aid (also known as a cryoprotectant).

[0047] Specifically, the lyophilization aid may be one or more selected from lactose, maltose, sucrose, trehalose, mannitol, sorbitol, maltitol, xylitol, and lactitol.

[0048] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention may further contain a local anesthetic.

[0049] Specifically, the local anesthetic may be selected from ambucaine, amolanone, amylocaine, benoxinate, benzocaine, betoxycaine, biphenamine, bupivacaine, butacaine, butamben, butanilicaine, butethamine, butoxycaine, articaine, chloroprocaine, cocaethylene, cocaine, cyclomethycaine, dibucaine, dimethysoquin, dimethocaine, diperodon, dicyclonine, ecgonidine, ecgonine, ethyl chloride, etidocaine, beta-eucaine, euprocin, fenalcomine, formocaine, hexylcaine, hydroxytetracaine, isobutyl p-aminobenzoate, leucinocaine mesylate, levoxadrol, lidocaine, mepivacaine, meprylcaine, metabutoxycaine, methyl chlorideone or more of chloride, myrtecaine, naepaine, octacaine, orthocaine, oxethazaine, parethoxycaine, phenacaine, phenol, piperocaine, piridocaine, polidocanol, pramoxine, prilocaine, procaine, propanocaine, proparacaine, propipocaine, propoxycaine, pseudococaine, pyrrocaine, ropivacaine, salicylalcohol, tetracaine, tolycaine, trimecaine, zolamine, and their salts.

[0050] The powder preparation for tissue repair treatment of the present invention can be a preparation having a powder property by drying through freeze-drying or other drying methods (such as spin drying, etc.).

[0051] In a specific embodiment, the powder preparation for tissue repair treatment of the present invention can be prepared by a method including the following steps: polymerizing the monomers of the above hydrophobic biocompatible polymer in the presence of the above hydrophilic biocompatible polymer to prepare a copolymer of the hydrophilic biocompatible polymer and the hydrophobic biocompatible polymer; and drying the prepared copolymer, but not limited thereto.

[0052] The step of polymerizing the monomers of the hydrophobic biocompatible polymer in the presence of the hydrophilic biocompatible polymer can be carried out according to known methods and conditions.

[0053] Specifically, the drying of the biocompatible copolymer can be carried out by freeze-drying or other drying methods (such as spin drying, etc.), more specifically by freeze-drying, but not necessarily limited thereto. The freeze-drying of the biocompatible copolymer can be carried out in the presence of the above freeze-drying aid, but not necessarily limited thereto.

[0054] The powder preparation for tissue repair treatment of the present invention is easily soluble in an aqueous medium at room temperature to form polymer particles (for example, particles showing an average diameter in the range of 20 nm to 5000 nm). After being introduced into the body, under the influence of the in-vivo environment, the particles self-assemble through the hydrophobic aggregation of the hydrophobic polymer to form large structures, are not phagocytosed by macrophages, and induce collagen, thus showing excellent tissue repair effects.

[0055] Without being limited to theory, the above characteristics of the powder preparation for tissue repair treatment of the present invention (that is, when a reference solution containing the powder preparation at a concentration of 7.5% by weight is diluted according to a series of dilution multiples that increase in sequence, (i) in the dilution multiple range of 4 to 8, the number of polymer particles per unit volume in the diluted solution is greater than or equal to the number of polymer particles per unit volume in the reference solution, (ii) in the dilution multiple range above 8, as the dilution multiple increases, the number of polymer particles per unit volume in the diluted solution decreases) can be considered to be due to the self-assembly characteristics of such polymer particles. That is, the powder preparation for tissue repair treatment of the present invention is considered that when prepared into a solution with a concentration equal to or higher than that of the reference solution, the polymer particles in the solution will self-assemble. As the solution is diluted, in the dilution multiple range of 4 to 8, the self-assembled particle clusters will split, and although diluted, the number of particles per unit volume of the solution will increase or remain unchanged. In the dilution multiple range above 8, when continued dilution is carried out, there are no particle clusters that will split, and as the dilution progresses, the number of particles per unit volume of the solution decreases. On the contrary, if the polymer particles in the solution are not self-assembled, as the dilution progresses, the number of particles per unit volume of the solution continuously decreases.

[0056] In the present invention, the tissue repair effect refers to the effect of restoring the tissue to its original state when necrosis, defects, etc. occur in the skin tissue due to reasons such as trauma, inflammation, aging, etc.

[0057] Therefore, in another aspect of the present invention, there is provided an injection composition for tissue repair treatment, which comprises: the powder preparation for tissue repair treatment of the present invention; and a pharmaceutically acceptable injection carrier.

[0058] The pharmaceutically acceptable injection carrier contained in the injection composition for tissue repair treatment of the present invention can use conventional carriers without limitation. For example, carriers selected from solutions containing hyaluronic acid (hyaluronic acid solution), distilled water for injection, physiological saline, 5% glucose, buffer solutions (such as phosphate buffer solution (PBS)), and combinations thereof can be used, but are not limited thereto.

[0059] In addition to the above components, the injection composition for tissue repair treatment of the present invention may further contain one or more conventional additives that can be used in injectable preparations.

[0060] Another aspect of the present invention provides a method for preparing an injection composition for tissue repair treatment, the method comprising mixing the powder preparation for tissue repair treatment of the present invention and a pharmaceutically acceptable injectable carrier at room temperature.

[0061] In the method for preparing the injection composition for tissue repair treatment of the present invention, "room temperature" means 1 - 30°C, 20 - 30°C, 22 - 28°C, and more specifically means 24 - 26°C (for example, 25°C).

[0062] The powder preparations of existing polymer products for tissue repair must be heated (for example, the temperature must be raised to between the melting point of the polymer and the boiling point of water) to be prepared in the form of an aqueous solution, but the powder preparation for tissue repair treatment of the present invention is also easily soluble in an aqueous medium at room temperature, so an injection composition in the form of an aqueous solution can be easily prepared and used at room temperature.

[0063] The following are examples of the present invention, but the scope of the present invention is not limited thereto.

[0064] [Examples]

[0065] Preparation Example 1: Preparation of Powder Preparation

[0066] To obtain a biocompatible copolymer with a target number average molecular weight (Mn) of 15,000 g / mol, in the presence of methoxypolyethyleneglycol (mPEG) with a number average molecular weight (Mn1) of 9,850 g / mol (measured by GPC) as a hydrophilic biocompatible polymer, D,L-lactide monomers were polymerized in the presence of a catalyst, and then the resulting copolymer was dried to obtain a powder preparation. As a result of GPC measurement, the number average molecular weight (Mn) of the prepared mPEG-poly(D,L-lactide) copolymer was 15,500 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number average molecular weight (Mn1) of mPEG as a hydrophilic polymer to the number average molecular weight (Mn2) of poly(D,L-lactide) as a hydrophobic polymer in the copolymer was 1.74.

[0067] Preparation Example 2: Preparation of Powder Preparation

[0068] A powder preparation was prepared by the same method as in Preparation Example 1, except that the target number-average molecular weight (Mn) of the biocompatible copolymer was set to 17,000 g / mol. As a result of GPC measurement, the number-average molecular weight (Mn) of the prepared mPEG-poly(D,L-lactide) copolymer was 17,200 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number-average molecular weight (Mn1) of mPEG as the hydrophilic polymer to the number-average molecular weight (Mn2) of poly(D,L-lactide) as the hydrophobic polymer in the copolymer was 1.34.

[0069] Preparation Example 3: Preparation of powder preparation

[0070] A powder preparation was prepared by the same method as in Preparation Example 1, except that the target number-average molecular weight (Mn) of the biocompatible copolymer was set to 20,000 g / mol. As a result of GPC measurement, the number-average molecular weight (Mn) of the prepared mPEG-poly(D,L-lactide) copolymer was 20,200 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number-average molecular weight (Mn1) of mPEG as the hydrophilic polymer to the number-average molecular weight (Mn2) of poly(D,L-lactide) as the hydrophobic polymer in the copolymer was 0.95.

[0071] Preparation Example 4: Preparation of powder preparation

[0072] In order to obtain a biocompatible copolymer with a target number-average molecular weight (Mn) of 30,000 g / mol, D,L-lactide monomer was polymerized in the presence of a catalyst in the presence of methoxypolyethylene glycol (mPEG) with a number-average molecular weight (Mn1) of 20,000 g / mol (measured by GPC) as the hydrophilic biocompatible polymer, and then the resulting copolymer was dried to obtain a powder preparation. As a result of GPC measurement, the number-average molecular weight (Mn) of the prepared mPEG-Poly(D,L-lactide) copolymer was 29,550 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number-average molecular weight (Mn1) of mPEG as the hydrophilic polymer to the number-average molecular weight (Mn2) of poly(D,L-lactide) as the hydrophobic polymer in the copolymer was 2.1.

[0073] Comparative Example 1

[0074] A powder preparation obtained by freeze-drying a product (Miracle L, manufactured by Dexlevo) composed of an mPEG-polycaprolactone copolymer was used as Comparative Example 1.

[0075] When the mPEG-polycaprolactone powder preparation of Comparative Example 1 was prepared into an aqueous solution by mixing at room temperature, due to poor dispersion stability, the polymer particles precipitated, making it difficult to analyze the particle size and reducing the reliability of the results. Therefore, water was added to the mPEG-polycaprolactone powder preparation and heated to a temperature of 80 °C, and then mixed to prepare a polymer colloidal aqueous solution for analysis.

[0076] Comparative Example 2: Preparation of powder preparation

[0077] A powder preparation was prepared by the same method as in Preparation Example 1, except that the target number average molecular weight (Mn) of the biocompatible copolymer was set to 12,000 g / mol. As a result of GPC measurement, the number average molecular weight (Mn) of the prepared mPEG-poly(D,L-lactide) copolymer was 12,350 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number average molecular weight (Mn1) of mPEG as the hydrophilic polymer to the number average molecular weight (Mn2) of poly(D,L-lactide) as the hydrophobic polymer in the copolymer was 2.8.

[0078] Comparative Example 3: Preparation of powder preparation

[0079] To obtain a biocompatible copolymer with a target number average molecular weight (Mn) of 25,000 g / mol, D,L-lactide monomers were polymerized in the presence of a catalyst in the presence of methoxy polyethylene glycol (mPEG) with a number average molecular weight (Mn1) of 20,000 g / mol (measured by GPC) as the hydrophilic biocompatible polymer, and then the resulting copolymer was dried to obtain a powder preparation. As a result of GPC measurement, the number average molecular weight (Mn) of the prepared mPEG-poly(D,L-lactide) copolymer was 25,650 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number average molecular weight (Mn1) of mPEG as the hydrophilic polymer to the number average molecular weight (Mn2) of poly(D,L-lactide) as the hydrophobic polymer in the copolymer was 3.5.

[0080] Comparative Example 4: Preparation of powder preparation

[0081] To obtain a biocompatible copolymer with a target number-average molecular weight (Mn) of 23,000 g / mol, in the presence of methoxypolyethylene glycol (mPEG) with a number-average molecular weight (Mn1) of 20,000 g / mol (measured by GPC) as a hydrophilic biocompatible polymer, D,L-lactide monomers were polymerized in the presence of a catalyst, and then the resulting copolymer was dried to obtain a powder formulation. As a result of GPC measurement, the number-average molecular weight (Mn) of the prepared mPEG-poly(D,L-lactide) copolymer was 23,500 g / mol, and the hydrophobic polymer molecular weight was calculated by subtracting the hydrophilic polymer molecular weight from the measured copolymer molecular weight. The ratio (Mn1 / Mn2) of the number-average molecular weight (Mn1) of mPEG as a hydrophilic polymer to the number-average molecular weight (Mn2) of poly(D,L-lactide) as a hydrophobic polymer in the copolymer was 5.7.

[0082] Comparative Example 5: Powder formulation of PCL nanoparticles

[0083] After purchasing polycaprolactone (PCL) nanoparticles with an average particle size of 200 nm (product number: PCL200-5ML-A) from Sigma-Aldrich, they were lyophilized to obtain a powder formulation.

[0084] Experimental Example 1: Polymer molecular weight analysis

[0085] Gel permeation chromatography (GPC) was performed under the conditions shown in the following table to measure the number-average molecular weight (Mn) of the polymers of Preparation Examples 1 to 4 and Comparative Examples 1 to 5, and the results are shown in Table 1 below.

[0086]

[0087] Experimental Example 2: Measurement of the number of polymer particles in a solution diluted with an aqueous medium according to the powder formulation

[0088] The formulations of Preparation Examples 1 to 4 and Comparative Examples 1 to 5 were respectively added to water for injection as an aqueous medium to prepare a solution with a concentration of 7.5% by weight (reference solution), and then the number of particles with an average diameter of 20 to 5000 nm present in the unit volume of the solution was measured according to flow cytometry (FlowCytometer, FC) using an APOGEE device (model name: A60 Micro Plus, manufacturer: APOGEE flow systems). The measurement conditions are described below.

[0089] (1) Equipment conditions and parameter settings

[0090] 1) Laser source: 405 nm (Violet) / 70 mW

[0091] 2) Flow rate: 0.75 μl / minute

[0092] 3) Injection volume: 100 μl

[0093] 4) PMT: SALS: 415 V / MALS: 400 V / LALS: 400 V (STD: Set to approximately 40000 ets / μl)

[0094] 5) Acquisition time: 120 s

[0095] 6) Thresholds: SALS - 5, MALS - 13, LALS - 3

[0096] (2) APOGEE STD measurement

[0097] Standard substance: Cat# 1527, APOGEE Mix 25 ml (PS80, 110, 500 & SI80, 240, 30, 590, 880, 1300 nm)

[0098] Lot: CAL0145

[0099] (3) Measurement and conversion of the number of Events

[0100] Measure the number of events in the same ROD area. If the measurement time is less than 120 seconds, determine the number of events by converting the measurement result to the measurement result for 120 seconds.

[0101] After that, dilute the reference solution with water for injection at dilution factors of 2, 4, 8, 16, 32, and 64. For each diluted solution, measure the number of particles with an average diameter of 20 to 5000 nm present in the unit volume of the solution by the same method.

[0102] The measured or converted number of events is shown as the number of particles in Tables 2 and 3 below.

[0103] In addition, when the number of particles in the reference solution is set to 100%, the number of particles in the diluted solutions is expressed as relative % in Tables 2 and 3 below, Figure 1 and Figure 2 the change in the number of particles in the diluted solutions with increasing dilution factor is expressed as relative % in

[0104]

[0105]

[0106]

[0107]

[0108] Experimental Example 3: Verification of the Efficacy of a Preparation for Tissue Repair through Animal Experiments

[0109] Verify the efficacy of the powder preparation according to the present invention for tissue repair through animal experiments. Animal experiments were conducted using 5-week-old Sprague-Dawley rats (Rat) (purchased from Orient Bio).

[0110] In the animal experiments, saline and the test substance were administered to both sides of each 5-week-old Sprague-Dawley rat. During the experiment, the breeding environment was set to a temperature of 24 ± 2°C, a relative humidity of 50 ± 10%, a light time of 12 hours, and free access to food.

[0111] The saline as the control group, the aqueous solution (10% concentration) of the powder preparations prepared in Preparation Examples 1 to 4 as the test substances dissolved in water for injection, the product of Comparative Example 1 (Miracle L, made into a 20% concentration), and the aqueous solutions (10% concentration) of the powder preparations of Comparative Examples 2 to 5 dissolved in water for injection were each injected at a constant rate of 100 μl, and then the experimental animals were sacrificed after 6 weeks. In order to evaluate the ability of new collagen biosynthesis, the skin tissue at the injection site of the sample ( Figure 3 the part indicated by the arrow in the upper figure) was stained with Masson's Trichrome (MT) and then the formation of collagen in the tissue was observed. The results are shown in Figure 3 ( Figure 3 the lower figure).

[0112] It was confirmed from Figure 3 that, compared with the comparative examples, especially compared with Miracle L, the aqueous solutions of the preparations of the preparation examples showed excellent formation of collagen. In particular, in the case of the preparation of Preparation Example 3, compared with the comparative examples, especially compared with Miracle L, significantly coarser and a large amount of collagen fibers were formed.

[0113] That is, it was confirmed that, compared with the comparative examples, especially compared with Miracle L, the powder preparation according to the present invention has the advantages of being easy to transport, store, and handle, being easily dissolved in an aqueous medium at room temperature, and more excellently inducing the formation of collagen when introduced into the body, thus showing a more excellent tissue repair effect.

Claims

1. A powder preparation for tissue repair treatment, characterized in that, the powder preparation comprises a biocompatible copolymer which is a copolymer of a hydrophilic biocompatible polymer and a hydrophobic biocompatible polymer, when a reference solution containing the powder preparation at a concentration of 7.5% by weight is diluted according to a series of dilution multiples increasing in sequence, in the dilution multiple range of 4 to 8, the number of polymer particles per unit volume in the diluted solution is greater than or equal to the number of polymer particles per unit volume in the reference solution, in the dilution multiple range above 16, as the dilution multiple increases, the number of polymer particles per unit volume in the diluted solution decreases.

2. The powder preparation for tissue repair treatment according to claim 1, wherein, in the dilution multiple range of 4 to 16, the number of polymer particles per unit volume in the diluted solution is greater than or equal to the number of polymer particles per unit volume in the reference solution.

3. The powder preparation for tissue repair treatment according to claim 1, wherein, in the dilution multiple range of 4 to 32, the number of polymer particles per unit volume in the diluted solution is greater than or equal to the number of polymer particles per unit volume in the reference solution.

4. The powder preparation for tissue repair treatment according to claim 1, wherein, in the dilution multiple range above 8, as the dilution multiple increases, the number of polymer particles per unit volume in the diluted solution decreases.

5. The powder preparation for tissue repair treatment according to claim 1, wherein, in the dilution multiple range above 4, as the dilution multiple increases, the number of polymer particles per unit volume in the diluted solution decreases.

6. The powder preparation for tissue repair treatment according to claim 1, wherein, the hydrophilic biocompatible polymer is selected from polyethylene glycol or its derivatives, polyvinylpyrrolidone, polyvinyl alcohol, polyacrylamide, and combinations thereof.

7. The powder preparation for tissue repair treatment according to claim 1, wherein, the hydrophobic biocompatible polymer is selected from polylactide, polyglycolide, poly(lactide - glycolide), polytropic acid, polycaprolactone, polydioxanone - 2 - one, polyamino acids, polyorthoesters, polyanhydrides, polycarbonates, and combinations thereof.

8. The powder preparation for tissue repair treatment according to claim 1, wherein, the hydrophilic biocompatible polymer is selected from polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), and combinations thereof, and the hydrophobic biocompatible polymer is selected from polylactide, polyglycolide, poly(lactide - glycolide), and combinations thereof.

9. The powder preparation for tissue repair treatment according to claim 1, wherein, the number - average molecular weight of the hydrophilic biocompatible polymer determined by gel permeation chromatography is 1000 g / mol to 30000 g / mol.

10. The powder preparation for tissue repair treatment according to claim 1, wherein, the number - average molecular weight of the hydrophobic biocompatible polymer determined by gel permeation chromatography is 500 g / mol to 30000 g / mol.

11. The powder preparation for tissue repair treatment according to claim 1, wherein, the number average molecular weight of the biocompatible copolymer determined by gel permeation chromatography is from 2,000 g / mol to 40,000 g / mol.

12. The powder preparation for tissue repair treatment according to claim 1, wherein, the ratio (Mn1 / Mn2) of the number average molecular weight (Mn1) of the hydrophilic biocompatible polymer to the number average molecular weight (Mn2) of the hydrophobic biocompatible polymer in the biocompatible copolymer is 2.5 or less.

13. The powder preparation for tissue repair treatment according to claim 1, wherein, the powder preparation further comprises a lyophilization aid.

14. The powder preparation for tissue repair treatment according to claim 1, wherein, the powder preparation further comprises a local anesthetic.

15. An injectable composition for tissue repair treatment, comprising: the powder preparation for tissue repair treatment according to any one of claims 1 to 14; and a pharmaceutically acceptable injectable carrier.

16. A method for preparing an injectable composition for tissue repair treatment, comprising mixing the powder preparation for tissue repair treatment according to any one of claims 1 to 14 and a pharmaceutically acceptable injectable carrier at room temperature.

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