Method for producing polymer microparticles, polymer microparticles, medical compositions containing the same, cosmetic compositions, medical supplies and cosmetic supplies

Abstract

The present invention relates to a method for producing polymeric microparticles that can achieve high crosslinking efficiency and production yield while also achieving excellent mechanical strength and stability, polymeric microparticles, and medical compositions, cosmetic compositions, medical supplies, and cosmetic supplies containing the same.

Description

Technical Field

[0001] [Cross - reference to Related Applications] This application claims the benefit of priority based on Korean Patent Application No. 10 - 2019 - 0120094 filed on September 27, 2019 and Korean Patent Application No. 10 - 2020 - 0124264 filed on September 24, 2020, and all the contents disclosed in the documents of the Korean patent applications are included as part of this specification.

[0002] The present invention relates to a method for producing polymer microparticles, polymer microparticles, a medical composition, a cosmetic composition, medical supplies, and cosmetic supplies that can achieve high cross - linking efficiency and production yield, as well as excellent mechanical strength and stability.

Background Art

[0003] As the fields of biopharmaceuticals and regenerative medicine expand, the requirements for cell mass culture technology capable of efficiently producing cells, tissues, microorganisms, etc. are increasing.

[0004] Adherent cells are cultured using microcarriers in a 3D bioreactor. By placing cells, a culture medium, and microcarriers in the bioreactor and stirring the culture medium to bring the cells and microcarriers into contact, the cells are attached to the surface of the microcarriers and cultured. At this time, the microcarriers used provide a high surface area ratio (surface area / volume) to which cells can attach and grow, so they are suitable for large - scale cell culture. However, when expanding and culturing adherent cells using microcarriers, the process of recovering cells inevitably accompanies the cell detachment process after the culture is completed. The cell detachment process induces cell detachment by using proteolytic enzymes or changing the temperature, but when such a detachment step is added, there are problems such as increased manufacturing costs, reduced economic efficiency, and the induction of cell damage.

[0005] The development of new materials and processes to solve this problem is steadily progressing, and in particular, in the case of cell therapy agents that inject cells into the body, there are efforts to ensure the biocompatibility of the microcarriers used to culture the cells and to eliminate the separation and purification process. In this case, particles that can withstand the stress applied by the fluids surrounding the carrier during the culture process and after injection into the body are required.

[0006] Furthermore, in transdermal drug delivery technology, where microcarriers containing drugs or bioactive substances are loaded onto microneedles for delivery, the microcarriers should be polymers suitable for application to the body and must have sufficient strength to prevent particle deformation during passage through the stratum corneum layer of the skin. Microcarriers that have stably penetrated the skin can then deliver the loaded drug locally or systemically, acting on the necessary lesions.

[0007] Hyaluronic acid, primarily used as a biocompatible substance, is composed of N-acetyl-D-glucosamine and D-glucuronic acid, and is a biopolymer in which the repeating units are linearly linked. It is abundant in the vitreous fluid of the eye, synovial fluid of joints, and rooster combs. Although hyaluronic acid is frequently used as an injectable substance due to its excellent biocompatibility and viscoelasticity, its use is limited because it decomposes easily in vivo or under acidic or alkaline conditions. Furthermore, when applied to microcarriers, it exhibits a surface negative charge within the biological pH range, significantly reducing cell adhesion.

[0008] On the other hand, gelatin is a polymer obtained by hydrolyzing collagen, a biological connective tissue, and is widely used as a scaffold for cell culture. While gelatin's cell adhesion properties can be used to collect and culture cells, its strength is weak and it exhibits phase transition characteristics with temperature. There is research underway to improve its physical properties by introducing functional groups using chemical methods.

[0009] Therefore, there is a need to develop microcarriers or polymer microparticles that are suitable for living organisms while possessing excellent physical properties such as physical strength and stability against heat and enzymes. [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] This invention provides a method for producing polymer microparticles that can achieve high crosslinking efficiency and production yield, as well as excellent mechanical strength and stability.

[0011] Furthermore, the present invention provides polymer microparticles produced by the above-described manufacturing method.

[0012] Furthermore, the present invention provides a medical composition containing the aforementioned polymer microparticles.

[0013] Furthermore, the present invention provides a cosmetic composition containing the aforementioned polymer microparticles.

[0014] Furthermore, the present invention provides a medical product containing the above-mentioned medical composition.

[0015] Furthermore, the present invention provides a beauty product containing the aforementioned beauty composition. [Means for solving the problem]

[0016] This specification provides a method for producing polymer microparticles, comprising the steps of: forming crosslinked particles by subjecting a mixture containing an emulsion of a biocompatible polymer and a first crosslinking agent to a primary crosslinking reaction; and extracting the crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent.

[0017] This specification also provides polymer microparticles comprising a polymer matrix in which biocompatible polymers are crosslinked via a crosslinking agent.

[0018] This specification also provides a medical composition comprising the polymer microparticles and a pharmaceutically active substance contained within the polymer microparticles.

[0019] This specification also provides a cosmetic composition comprising the polymer microparticles and a cosmetic active substance contained within the polymer microparticles.

[0020] This specification also provides a medical product comprising the medical composition.

[0021] This specification also provides a cosmetic product comprising the cosmetic composition.

[0022] Hereinafter, the method for producing polymer microparticles, the polymer microparticles, the medical composition, the cosmetic composition, the medical product, and the cosmetic product according to specific embodiments of the invention will be described in more detail.

[0023] Unless explicitly mentioned in this specification, the technical terms are merely for referring to specific examples and are not intended to limit the present invention.

[0024] As used in this specification, the singular form also includes the plural form unless the context clearly indicates the contrary meaning.

[0025] The meaning of "comprising" as used in this specification is not to exclude the presence or addition of other specific characteristics, regions, integers, steps, operations, elements, and / or components while specifying a particular characteristic, region, integer, step, operation, element, and / or component.

[0026] And terms containing ordinal numbers such as "first" and "second" in this specification are used for the purpose of distinguishing one component from another and are not limited by the ordinal numbers. For example, within the scope of the rights of the present invention, the first component may also be named the second component, and similarly, the second component may also be named the first component.

[0027] In this specification, the (co)polymer means either a polymer or a copolymer. The polymer means a homopolymer consisting of a single repeating unit, and the copolymer means a composite polymer containing two or more repeating units.

[0028] In this specification, the molecular weight of the polymer used follows the measurement method of the manufacturing company, and the method must not deviate from the normal GPC measurement method. For example, in this specification, the weight average molecular weight means the molecular weight measured by light scattering methods or viscosity methods.

[0029] Since the present invention can be subjected to various modifications and can have various forms, specific examples are illustrated and described in detail below. However, this is not intended to limit the present invention to a specific disclosed form, and it should be understood as including all modifications, equivalents, or alternatives included in the above idea and technical scope. [[ID=??]] [[ID=??]]

[0030] [[ID=??]] In this specification, the microparticles mean that the cross-section of the particles is circular or elliptical, and the ratio of the short axis to the long axis (sphericity) of the particles is in the range of 0.7 to 1.0. The lengths of the short axis and the long axis of the particles can be derived by taking an optical photograph of the particles and calculating the average value of any 30 to 100 particles in the optical photograph.

[0031] In this specification, the particle size (Dn) means the particle size at the n volume% point of the cumulative distribution of particle numbers according to the particle size. That is, D50 is the particle size at the 50% point of the cumulative distribution of particle numbers when the particle sizes are cumulatively arranged in ascending order, D90 is the particle size at the 90% point of the cumulative distribution of particle numbers according to the particle size, and D10 is the particle size at the 10% point of the cumulative distribution of particle numbers according to the particle size.

[0032] Note: There seem to be some incorrect tag references ([[ID=??]]) in the original text which might be a formatting error. I've translated the text as accurately as possible based on the provided content.The aforementioned Dn can be measured using the laser diffraction method. Specifically, after dispersing the powder to be measured in a dispersion medium, it is introduced into a commercially available laser diffraction particle size analyzer (Horiba LA-960), and the difference in diffraction patterns due to particle size is measured as the particles pass through the laser beam to calculate the particle size distribution. By calculating the particle diameter at the points where the cumulative distribution of particle number by particle size in the measuring device reaches 10%, 50%, and 90%, D10, D50, and D90 can be measured. More specifically, in this specification, particle size refers to D50.

[0033] In this specification, an emulsion refers to a mixed phase in which one or more immiscible liquids, such as an oil phase or an aqueous phase, are dispersed as fine particles (dispersed phase) in another liquid (dispersion medium). Emulsions are usually classified into macroemulsions, microemulsions, and nanoemulsions depending on the particle size of the dispersed phase.

[0034] The present invention will be described in more detail below. 1. Method for producing polymer microparticles

[0035] According to one embodiment of the invention, a method for producing polymer microparticles is provided, comprising the steps of: forming crosslinked particles by causing a primary crosslinking reaction of a mixture containing a biocompatible polymer in an emulsion state and a first crosslinking agent; and extracting the crosslinked particles and causing a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent.

[0036] The term "emulsion" refers to a water-in-oil emulsion, an oil-in-water emulsion, an oil-in-oil emulsion, or a multiple emulsion. Specifically, the emulsion in the above embodiment refers to a water-in-oil emulsion.

[0037] Conventional polymer microparticles are formed by directly adding particles to an organic solvent phase to carry out a crosslinking reaction. However, the morphology of these particles is greatly affected by the concentration of the polymer and the amount of crosslinking agent. In particular, when the concentration of the polymer is low or the amount of crosslinking agent is low, the morphology of the particles is not maintained, making it difficult to form microparticles.

[0038] Therefore, the inventors completed the invention by experimentally confirming that, as in the method for producing polymer microparticles of the above embodiment, by performing a crosslinking reaction in two stages, the crosslinking density can be maximized with the same amount of crosslinking agent, while at the same time the mechanical strength and stability of the microparticles are significantly improved.

[0039] In particular, experiments confirmed that by performing the secondary crosslinking reaction in solid phase, spherical particles can be produced even with a small amount of crosslinking agent, thereby improving the production yield.

[0040] Specifically, the biocompatible polymer refers to a polymer that can be directly injected into the human body for the effective delivery of pharmaceuticals applied to the human body. Specifically, the biocompatible polymers include hyaluronic acid (HA), carboxymethyl cellulose (CMC), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), chitosan, polylysine, collagen, gelatin, and carboxymethyl chitin. Chitin, fibrin, agarose, pullulan, polylactide, polyglycolide (PGA), polylactide-glycolide copolymer (PLGA), polyanhydride, polyorthoester, polyetherester, polycaprolactone, polyesteramide, poly(butyric acid), poly(valeric acid), polyurethane, polyacrylate, ethylene-vinyl acetate polymer, acrylic-substituted cellulose acetate, non-degradable polyurethane, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinylimidazole), chlorosulfonate polyolefin (chlorosulfonate It may be one or more polymers selected from the group consisting of polyolefins, polyethylene oxide, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), hydroxypropylcellulose (HPC), cyclodextrin, poly(N-isopropylamide) (PNIPAam), poloxamer, and polyacrylic acid copolymers and derivatives thereof.

[0041] More specifically, the biocompatible polymer may be hyaluronic acid (HA), gelatin, or a mixture of hyaluronic acid and gelatin.

[0042] In this specification, hyaluronic acid may mean both hyaluronic acid itself and hyaluronic acid salts. Therefore, an aqueous solution of hyaluronic acid may be a concept that includes an aqueous solution of hyaluronic acid, an aqueous solution of hyaluronic acid salts, and an aqueous solution of a mixture of hyaluronic acid and hyaluronic acid salts. The hyaluronic acid salts may be inorganic salts such as sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, and cobalt hyaluronate, and organic salts such as tetrabutylammonium hyaluronate and mixtures thereof.

[0043] In one embodiment of the invention, the molecular weight of hyaluronic acid is not particularly limited, but it is preferably 500 g / mol or more and 5,000,000 g / mol or less in order to achieve diverse physical properties and biocompatibility.

[0044] In this specification, gelatin means a protein obtained by treating collagen derived from animals with an acid or alkali and subsequently extracting it.

[0045] In one embodiment of the invention, the molecular weight of gelatin is not particularly limited, but it is preferably 100,000 g / mol or more and 5,000,000 g / mol or less in order to achieve diverse physical properties and biocompatibility.

[0046] In one embodiment of the above invention, the first crosslinking agent is not significantly limited in its examples. Specifically, the first crosslinking agent is butanediol diglycidyl ether (1,4-butandiol diglycidyl ether: BDDE), ethylene glycol diglycidyl ether (ethylene glycol diglycidyl ether: EGDGE), hexanediol diglycidyl ether (1,6-hexanediol diglycidyl ether), propylene glycol diglycidyl ether (propylene glycol diglycidyl ether), polypropylene glycol diglycidyl ether (polypropylene glycol diglycidyl ether), polytetramethylene glycol diglycidyl ether (polytetramethylene glycol diglycidyl ether), neopentyl glycol diglycidyl ether (neopentyl glycol diglycidyl ether), polyglycerol polyglycidyl ether (polyglycerol polyglycidyl ether), diglycerol polyglycidyl ether (diglycerol polyglycidyl ether). The first crosslinking agent may include one selected from the group consisting of ether, tri-methylpropane polyglycidyl ether, 1,2-(bis(2,3-epoxypropoxy)ethylene), pentaerythritol polyglycidyl ether, sorbitol polyglycidyl ether, divinyl sulfone, and glutaraldehyde. More specifically, the first crosslinking agent may be 1,4-butandiol diglycidyl ether (BDDE).

[0047] In one embodiment of the invention, the step of forming crosslinked particles by a primary crosslinking reaction of a mixture containing the biocompatible polymer in emulsion state and a first crosslinking agent may include the steps of: adding an aqueous solution in which the biocompatible polymer is dissolved to a hydrophobic solvent to form an emulsion; and adding the first crosslinking agent to a mixed solution containing the emulsion.

[0048] In other words, the primary crosslinking reaction may be a liquid-phase reaction carried out by adding a first crosslinking agent to an emulsion containing the biocompatible polymer.

[0049] Furthermore, in the step of forming crosslinked particles by performing a primary crosslinking reaction on a mixture containing the biocompatible polymer in emulsion state and the first crosslinking agent, the first crosslinking agent may be included in an amount of 30 to 300 parts by weight, 40 to 250 parts by weight, or 50 to 200 parts by weight per 100 parts by weight of the biocompatible polymer. The method for producing polymer microparticles according to one embodiment of the invention, as described above, allows for the production of polymer microparticles that achieve sufficient mechanical strength and degree of sphericity even when the first crosslinking agent is added in an amount of 30 to 300 parts by weight per 100 parts by weight of the biocompatible polymer by performing a secondary crosslinking reaction after the primary crosslinking reaction.

[0050] If the amount of the first crosslinking agent added is less than 30 parts by weight per 100 parts by weight of the biocompatible polymer, the degree of crosslinking is low and particle formation is difficult. If it is added in amounts exceeding 300 parts by weight, technical problems may arise in which an excess amount of unreacted crosslinking agent is not purified and remains in the crosslinked particles.

[0051] On the other hand, according to one embodiment of the invention, in the step of extracting the crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent, the crosslinked particles and the second crosslinking agent undergo the crosslinking reaction in a solid phase.

[0052] In other words, the method for producing polymer microparticles of the present invention may include a secondary crosslinking reaction carried out from an alkaline organic solvent phase to a solid phase. Furthermore, the secondary crosslinking reaction carried out in the solid phase refers to a secondary crosslinking reaction carried out in a more aggregated state after dehydration of the swollen polymer particles. When a secondary crosslinking reaction carried out from an organic solvent phase to a solid phase is included, as in one embodiment of the invention, that is, when a secondary crosslinking reaction carried out in a more aggregated state after dehydration of the swollen polymer particles is included, a superior crosslinking efficiency can be achieved compared to cases where this is not included, and microparticles with high strength can be obtained.

[0053] In one embodiment of the invention, the crosslinked particles are recovered using a mesh sieve with a mesh size of 30 μm or larger. After recovering the crosslinked particles formed by the primary crosslinking reaction in this manner, the particles are redispersed in an organic solvent and a second crosslinking agent is added, allowing the secondary crosslinking reaction to be carried out in a more aggregated solid phase.

[0054] In one embodiment of the above invention, the second crosslinking agent is not significantly limited in its examples. Specifically, the second crosslinking agent is butanediol diglycidyl ether (1,4-butandiol diglycidyl ether: BDDE), ethylene glycol diglycidyl ether (EGDGE), hexanediol diglycidyl ether (1,6-hexanediol diglycidyl ether), propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, diglycerol polyglycidyl ether, glycerol polyglycidyl The second crosslinking agent may include one selected from the group consisting of ether, tri-methylpropane polyglycidyl ether, 1,2-(bis(2,3-epoxypropoxy)ethylene), pentaerythritol polyglycidyl ether, sorbitol polyglycidyl ether, divinyl sulfone, and glutaraldehyde. More specifically, the second crosslinking agent may be 1,4-butandiol diglycidyl ether (BDDE).

[0055] That is, in one embodiment of the invention, the first crosslinking agent and the second crosslinking agent are each independently butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE), hexanediol diglycidyl ether (1,6-hexanediol diglycidyl ether), propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, diglycerol polyglycidyl ether, and glycerol polyglycidyl ether. The crosslinking agent may include one selected from the group consisting of ether, tri-methylpropane polyglycidyl ether, 1,2-(bis(2,3-epoxypropoxy)ethylene), pentaerythritol polyglycidyl ether, sorbitol polyglycidyl ether, divinyl sulfone, and glutaraldehyde, and preferably both the first and second crosslinking agents are 1,4-butandiol diglycidyl ether (BDDE).

[0056] Furthermore, the second crosslinking agent may be included in an amount of 30 to 300 parts by weight, 40 to 250 parts by weight, or 50 to 200 parts by weight per 100 parts by weight of the biocompatible polymer. In one embodiment of the invention, the method for producing polymer microparticles involves performing a secondary crosslinking reaction after the primary crosslinking reaction, as described above, thereby adding 30 to 300 parts by weight of the second crosslinking agent per 100 parts by weight of the biocompatible polymer to produce polymer microparticles that achieve sufficient mechanical strength and degree of spheroidization.

[0057] When the second crosslinking agent is added in an amount of less than 30 parts by weight per 100 parts by weight of the biocompatible polymer, the degree of crosslinking is low and particle formation is difficult. When added in amounts exceeding 300 parts by weight, technical problems may arise in which an excess amount of unreacted crosslinking agent is not removed and remains in the crosslinked particles.

[0058] On the other hand, in the step of extracting the crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent, the organic solvent is ethanol, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylcaprolactam, 2-pyrrolidone, N-ethylpyrrolidone, N-vinylpyrrolidone, dimethyl sulfoxide, tetramethylurea, pyridine, dimethyl sulfone, hexamethyl sulfoxide, gamma-butyrolactone, 3-methoxy-N,N-dimethylpropanamide, 3-ethoxy-N,N-dimethylpropanamide, 3-butoxy-N,N-dimethylpropanamide, 1,3-dimethylimidazolidinone, ethyl amyl ketone, methyl nonyl ketone, methyl ethyl ketone, methyl It may be one selected from the group consisting of isoamyl ketone, methyl isopropyl ketone, cyclohexanone, ethylene carbonate, propylene carbonate, diglyme, 4-hydroxy-4-methyl-2-pentanone, ethylene glycol monomethyl ether, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether, ethylene glycol monoethyl ether acetate, ethylene glycol monopropyl ether, ethylene glycol monopropyl ether acetate, ethylene glycol monoisopropyl ether, ethylene glycol monoisopropyl ether acetate, ethylene glycol monobutyl ether, and ethylene glycol monobutyl ether acetate.

[0059] Furthermore, in the step of extracting the crosslinked particles and carrying out a secondary crosslinking reaction in an organic solvent phase containing the second crosslinking agent, the organic solvent containing the second crosslinking agent may be an alkaline mixed solvent. That is, the secondary crosslinking reaction of the present invention can be carried out in an organic solvent phase in which an alkaline aqueous solution is mixed with ethanol, an organic solvent. Examples of the alkaline solution are not greatly limited, but could be, for example, an aqueous solution of sodium hydroxide.

[0060] The fact that the organic solvent containing the second crosslinking agent is an alkaline mixed solvent allows for a nucleophilic substitution reaction (S N By creating an environment favorable to the reaction, the efficiency of the crosslinking reaction can be increased.

[0061] 2. Molecular microparticles According to one embodiment of the invention, polymer microparticles are provided, comprising a polymer matrix in which biocompatible polymers are crosslinked via a crosslinking agent.

[0062] The inventors conducted research on polymer microparticles and, as described above, experimentally confirmed that by undergoing a primary crosslinking reaction in a swollen emulsion followed by a secondary crosslinking reaction in a dehydrated and aggregated solid phase, the crosslinking density can be maximized with the same amount of crosslinking agent, while simultaneously significantly improving the mechanical strength and stability of the microparticles, thus completing the invention.

[0063] In particular, by carrying out the secondary crosslinking reaction in a more aggregated solid phase during the two-stage crosslinking reaction, it was confirmed through experiments that spherical particles could be produced even with a small amount of crosslinking agent, and the production yield was improved, thus completing the invention.

[0064] Specifically, the polymer matrix may include a first crosslinked region in which a biocompatible polymer is crosslinked via a first crosslinking agent, and a second crosslinked region in which a biocompatible polymer is crosslinked via a second crosslinking agent.

[0065] The first crosslinked region refers to a crosslinked region formed by a primary crosslinking reaction between an emulsion containing a biocompatible polymer and a first crosslinking agent, while the second crosslinked region refers to a crosslinked region formed by a secondary crosslinking reaction between the emulsion containing a biocompatible polymer and the second crosslinking agent, since no primary crosslinking reaction occurs between the emulsion and the first crosslinking agent, and a crosslinked region formed by an additional secondary crosslinking reaction between the first crosslinked region and the second crosslinking agent.

[0066] On the other hand, the polymer microparticles may have an average diameter in distilled water of 1 μm to 650 μm, 100 μm to 600 μm, 200 μm to 550 μm, or 300 μm to 500 μm. However, when the average diameter of the polymer microparticles falls within the range described above, they exhibit superior cell adhesion and culture performance.

[0067] Furthermore, the polymer microparticles may have a degree of sphericity of 0.9 to 1.0, 0.95 to 1.0, 0.96 to 0.99, or 0.97 to 0.99.

[0068] The degree of sphericity can be obtained by taking optical photographs of polymer microparticles and calculating the average value of 30 to 100 arbitrary particles in the optical photographs.

[0069] Furthermore, the polymer microparticles may have a swelling degree of 55 or less, 0.1 to 55 or less, 0.1 to 50 or less, 0.2 to 50 or less, 0.5 to 30 or less, or 0.8 to 18 or less, according to the following formula 1.

number

[0070] If the swelling degree of the polymer microparticles exceeds 55, technical problems may arise such as low crosslinking density, poor mechanical strength, and reduced stability.

[0071] Furthermore, the polymer microparticles may have an average compressive strength of 0.1 mN or more, 0.1 mN to 100 mN, 0.11 mN to 100 mN, 1 mN to 100 mN, 1 mN to 90 mN, or 1 mN to 60 mN when deformed to 50% of the average particle diameter.

[0072] The average compressive strength may be the value obtained by dividing the compressive force when n polymer microparticles are deformed to 50% of their average diameter by n.

[0073] For example, in this specification, the average compressive strength may be the value obtained by dividing the compressive force when the polymer microparticles 30 are deformed to 50% of their average diameter by 30.

[0074] If the average compressive strength of the polymer microparticles is less than 0.1 mN, technical problems may arise where the mechanical strength of the polymer microparticles is poor, and the cell culture capacity is significantly reduced.

[0075] 3. Medical Compositions Another embodiment of the present invention provides a medical composition comprising polymer microparticles of the other embodiment and a pharmaceutically active substance contained within the polymer microparticles. The details relating to the polymer microparticles may include all of the details described above in the other embodiment.

[0076] The pharmaceutically active substance may exist contained within the polymer microparticles.

[0077] The examples of the aforementioned pharmaceutically active substances are not particularly limited, and depending on the application of the polymer microparticles of the above embodiment, any active substance suitable for the application can be applied without limitation.In other words, specific examples of the aforementioned pharmaceutically active substances are not limited to ampetaminil, arecolin, atrophine, bupranolol, buprenorphine, capsaicin, carisoprodol, chlorpromazine, cyclopirox olamine, cocaine, desipramine, dyclonine, epinephrine, ethosuximide, floxetine, hydromorphine, imipramine, lidocaine, methamphetamine, and melproic acid. Drugs selected from the group consisting of (1) methylphenidate, morphine, oxybutynin, nadolol, nicotine, nitroglycerin, pindolol, prilocaine, procaine, propanolol, rivastigmine, scopolamine, selegiline, tulobuterol, valproic acid, donepezil, etc., as well as EPO (Erythropoietin), human growth hormone (hGH), exenatide, GLP-1 (Glucagon-like peptide-1), insulin, CSF (Granulocyte colony-stimulating) This includes peptide or protein-based drugs selected from the group consisting of factors, estrogen, progesterone, paratyloid hormone (PTH), etc., and all pharmacologically proven pharmacologically active substances can be applied without restriction.

[0078] The amount of the aforementioned pharmaceutical active substance added is also not significantly limited, and its content can be used without restriction depending on the application and target. For example, the active substance can be included in amounts of 0.0001 parts by weight or more and 1000 parts by weight or less per 100 parts by weight of the polymer microparticles, without any restrictions on the amount compared to the polymer microparticles.

[0079] 4. Beauty composition According to yet another embodiment of the present invention, a cosmetic composition is provided comprising polymer microparticles of the other embodiment and a cosmetic active substance contained within the polymer microparticles. The details relating to the polymer microparticles may include all of the details described above in the other embodiment.

[0080] The aforementioned cosmetically effective substance can exist in a state contained within the polymer microparticles.

[0081] The examples of the aforementioned cosmetic active substances are not limited, and depending on the application of the polymer microparticles in the embodiment described above, any active substance suitable for that application can be applied without restriction. In other words, specific examples of the aforementioned cosmetic active substances are not limited, and include natural extracts, proteins, vitamins, enzymes, antioxidants, etc., and all substances whose cosmetic effects have been proven can be applied without restriction.

[0082] The amount of the cosmetic active substance added is also not significantly limited, and its content can be used without restriction depending on the application and target. For example, the cosmetic active substance can be included in an amount of 0.0001 parts by weight or more and 1000 parts by weight or less per 100 parts by weight of the polymer microparticles, without any restrictions on the amount compared to the polymer microparticles.

[0083] 5. Medical supplies According to yet another embodiment of the present invention, a medical product comprising the medical composition of the other embodiment is provided. The content relating to the medical composition may include all of the content described above in the other embodiment.

[0084] The examples of the aforementioned medical products are not limited, but are suitable for cases where they must be inserted into the body or maintain their strength for a long period of time in order to realize the characteristics of the present invention. Examples include internal prostheses, implantable drug delivery systems, transdermal patches, wound healing agents, and the like.

[0085] 6.Beauty supplies According to yet another embodiment of the present invention, the present invention provides a beauty product comprising the beauty composition of the other embodiment. The content relating to the beauty composition may include all of the content described above in the other embodiment.

[0086] The examples of the aforementioned beauty products are not limited, but to realize the characteristics of the present invention, examples include beauty creams, lotions, hair gels, masks, etc.

[0087] The structure of the beauty pack is not broadly limited, but may include, for example, a support and a cosmetic active substance delivery layer formed on the support and containing polymer microparticles of the other embodiments. Examples of the support include woven fabric, nonwoven fabric, silicone, polyethylene terephthalate, polyethylene, polypropylene, polyurethane, metal mesh, polyester, and the like. [Effects of the Invention]

[0088] According to the present invention, the present invention can provide a method for producing polymer microparticles that can achieve high crosslinking efficiency and production yield, as well as excellent mechanical strength and stability, and polymer microparticles using the same. [Modes for carrying out the invention]

[0089] The invention will be described in more detail by the following embodiments. However, the following embodiments are for illustrative purposes only, and the content of the invention is not limited by the following embodiments.

[0090] Example 1: Production of polymer microparticles 0.15 g of hyaluronic acid (weight-average molecular weight: 500 kDa, manufacturer: SK Bioland) and 1 g of gelatin (gel strength: 300 g Bloom, manufacturer: Sigma, product name: G2500) were dissolved in distilled water at 3 wt.% and 20 wt.%, respectively, to prepare a solution. These two solutions were then mixed in a 1:1 volume ratio. This mixed solution (11.15 g) was mixed with liquid paraffin solution (40 g) to prepare a mixture containing a water-in-oil (W / O) microemulsion. Subsequently, 1.1 g (1 ml) of 1,4-butanediol diglycidyl ether (BDDE) was added to 51.15 g of the mixture as a crosslinking agent, and the crosslinking reaction was carried out at room temperature for 5 days. After that, crosslinked particles were produced by washing in the order of acetone, dichloromethane, and distilled water. At this time, the cross-linked particles produced were recovered using a mesh sieve with a mesh size of 45 μm.

[0091] A solution was prepared by adding 1.1 g (1 ml) of 1,4-butanediol diglycidyl ether (BDDE) to a mixture of 31.56 g of ethanol and 9.88 g of 0.1 N NaOH aqueous solution. The cross-linked particles were added to this solution and the cross-linking reaction was carried out at room temperature for 3 days to produce polymer microparticles. The produced particles were washed with ethanol and then distilled water, and the produced cross-linked particles were recovered using a sieve with a mesh size of 45 μm. The recovered cross-linked particles were passed through a sieve with a mesh size of 300 μm, and the remaining cross-linked particles were analyzed.

[0092] Example 2: Production of polymer microparticles High-molecular-weight microparticles were produced in the same manner as in Example 1, except that 1 g of gelatin was added instead of hyaluronic acid.

[0093] Comparative Example 1: Production of Polymer Microparticles Hyaluronic acid (weight-average molecular weight: 500 kDa, manufacturer: SK Bioland) 0.15 g and gelatin (gel strength: 300 g Bloom, manufacturer: Sigma, product name: G2500) 1 g were dissolved in distilled water at 3 wt.% and 20 wt.% concentrations, respectively, to prepare a solution. These two solutions were then mixed in a 1:1 volume ratio. This mixed solution (11.15 g) was mixed with liquid paraffin solution (40 g) to prepare a mixture containing a water-in-oil (W / O) microemulsion. Subsequently, 1.1 g (1 ml) of 1,4-butanediol diglycidyl ether (BDDE) was added to 51.15 g of the mixture as a crosslinking agent, and the crosslinking reaction was carried out at room temperature for 5 days.

[0094] Subsequently, 1.1 g (1 ml) of 1,4-butanediol diglycidyl ether (BDDE) was added to the mixture as a crosslinking agent, and the crosslinking reaction was carried out at room temperature for 3 days to produce polymer microparticles. The produced particles were washed in acetone, dichloromethane, and distilled water in that order, and the produced crosslinked particles were recovered using a sieve with a mesh size of 45 μm. The recovered crosslinked particles were passed through a sieve with a mesh size of 300 μm, and the remaining crosslinked particles were analyzed.

[0095] Comparative Example 2: Production of Polymer Microparticles Hyaluronic acid (weight-average molecular weight: 500 kDa, manufacturer: SK Bioland) 0.15 g and gelatin (gel strength: 300 g Bloom, manufacturer: Sigma, product name: G2500) 1 g were dissolved in distilled water at 3 wt.% and 20 wt.% concentrations, respectively, to prepare a solution. These two solutions were then mixed in a 1:1 volume ratio. This mixed solution (11.15 g) was mixed with liquid paraffin solution (40 g) to prepare a mixture containing a water-in-oil (W / O) microemulsion. Subsequently, 1.1 g (1 ml) of 1,4-butanediol diglycidyl ether (BDDE) was added to 51.15 g of the mixture as a crosslinking agent, and the crosslinking reaction was carried out at room temperature for 5 days. The manufactured particles were washed with acetone, dichloromethane, and distilled water in that order, and then the manufactured cross-linked particles were recovered using a sieve with a mesh size of 45 μm.

[0096] The cross-linked particles were added to a mixture of 31.56 g of ethanol and 9.88 g of 0.1 N NaOH aqueous solution, and the cross-linking reaction was carried out at room temperature for 3 days to produce polymer microparticles. The produced particles were washed with ethanol and then distilled water, and the produced cross-linked particles were recovered using a sieve with a mesh size of 45 μm. The recovered cross-linked particles were passed through a sieve with a mesh size of 300 μm, and the remaining cross-linked particles were analyzed.

[0097] Comparative Example 3: Production of Polymer Microparticles Hyaluronic acid (weight-average molecular weight: 500 kDa, manufacturer: SK Bioland) 0.15 g and gelatin (gel strength: 300 g Bloom, manufacturer: Sigma, product name: G2500) 1 g were dissolved in distilled water at 3 wt.% and 20 wt.% concentrations, respectively, to prepare a solution. These two solutions were then mixed in a 1:1 volume ratio. This mixed solution (11.15 g) was mixed with liquid paraffin solution (40 g) to prepare a mixture containing a water-in-oil (W / O) microemulsion. Subsequently, 1.1 g (1 ml) of 1,4-butanediol diglycidyl ether (BDDE) was added to 51.15 g of the mixture as a crosslinking agent, and the crosslinking reaction was carried out at room temperature for 5 days. The manufactured particles were washed in the order of acetone, dichloromethane, and distilled water, and the manufactured cross-linked particles were recovered using a sieve with a mesh size of 45 μm. The recovered cross-linked particles were then passed through a sieve with a mesh size of 300 μm, and the remaining cross-linked particles were analyzed.

[0098] Experimental example: Measurement of the physical properties of polymer microparticles The polymer microparticles produced in the above examples and comparative examples were evaluated for their swelling degree, spheroidization degree, strength, cell culture suitability, and stability using the following methods.

[0099] 1. Average diameter and degree of swelling The average diameter of the polymer microparticles in the above examples and comparative examples was measured, and the degree of swelling was calculated based on this.

[0100] The degree of particle swelling according to the present invention was calculated using the following formula 1.

number

[0101] In this case, a swelling degree value closer to 0 means that no swelling occurs, while a larger value means that the particles swell and become larger.

[0102] The average diameter in distilled water was measured using a laser particle size analyzer (Horiba, Partica LA-960), and the average diameter after drying was calculated from the diameters of any 30 particles in SEM (Hitach, S-4800) images of the dried microparticles.

[0103] 2. Spheroidization degree Optical (Olympus, BX53) photographs were taken of the polymer microparticles of the above examples and comparative examples, and the degree of spheroidization was calculated based on them.

[0104] The degree of sphericity according to the present invention was calculated as the average of the ratio (aspect ratio) of the longest diameter to the shortest diameter of any 30 particles in an optical photograph. In this case, the closer the sphericity value is to 1, the closer it is to a sphere.

[0105] 3.Strength The strength of the polymer microparticles in the above examples and comparative examples was measured using a texture analyzer. Thirty microparticles swollen with distilled water were placed in a single layer in the area below a flat cylindrical probe fitted with a 5N load cell. The initial trigger force was set to 1 mN, and the particles were compressed at a speed of 1 mm / s. The force at which the average diameter of the particles was deformed to 50% of their original diameter was defined as the compressive force.

[0106] The average compressive strength is the value obtained by dividing the compressive force by 30, which is the number of microparticles being measured.

[0107] 4.Cell culture compatibility Cell culture medium was placed in a 6-well plate, and cells were added along with high-molecular-weight microparticles. Cells were then cultured using the plate-rocking method. During this time, the culture medium temperature was maintained at 37°C, and the number of cells cultured with high-molecular-weight microparticles was confirmed after 3 days of incubation.

[0108] At this time, cell culture compatibility was evaluated according to the following criteria. Suitable: When the number of cultured cells is 150% or more of the number of cells introduced. Failure to meet the requirement: The number of cultured cells is less than 150% of the number of cells introduced.

[0109] 5. Stability The sterilization stability of high-molecular-weight microparticles contained in phosphate-buffered saline solution and the particle stability during long-term culture were evaluated using a high-temperature, high-pressure sterilizer (Autoclave) according to the following criteria. Suitable for: When the weight loss rate of dried polymer microparticles before and after use in a high-temperature, high-pressure sterilizer (Autoclave) is 10% or less. Non-compliant: When the weight loss rate of dried polymer microparticles exceeds 10% before and after use of a high-temperature, high-pressure sterilizer (Autoclave).

[0110] [Table 1]

[0111] As shown in Table 1 above, the polymer microparticles of the examples produced by the manufacturing method of the present invention, which includes a step of recovering the particles after primary crosslinking and performing secondary crosslinking, not only showed that the number of cultured cells was 150% or more of the number of cells initially introduced, making them suitable for cell culture, but also showed a weight reduction rate of 10% or less for the dried polymer microparticles before and after high-temperature, high-pressure sterilization (Autoclave) treatment, confirming their suitability for sterilization treatment and long-term culture.

[0112] Furthermore, the polymer microparticles in the examples showed a high degree of sphericity with a degree of sphericity of 0.9 or higher, while simultaneously exhibiting excellent crosslinking with a swelling degree of 0.82 to 17.19. In addition, they demonstrated excellent mechanical properties with an average compressive strength of 1.17 mN or higher, confirming that the particles possessed a high crosslinking density.

[0113] In other words, it was confirmed that the polymer microparticles of the examples produced by the manufacturing method of the present invention, which includes a step of recovering the particles after primary crosslinking and performing secondary crosslinking, achieve excellent crosslinking density and mechanical properties while being suitable for cell culture, sterilization, and long-term culture.

[0114] On the other hand, in Comparative Example 1, where the polymer microparticles were not recovered after primary crosslinking but immediately subjected to secondary crosslinking, the weight loss rate of the dried polymer microparticles before and after high-temperature, high-pressure sterilization (Autoclave) treatment exceeded 10%, indicating that they were not only unsuitable for sterilization and long-term cultivation, but also showed a swelling degree of 56.51, confirming that the degree of crosslinking was inferior compared to the examples of the present application. Furthermore, the average compressive strength was 0.9 mN, indicating inferior mechanical properties, confirming that the particles had a low crosslinking density.

[0115] Furthermore, the polymer microparticles of Comparative Example 2, which were recovered after primary crosslinking and subjected only to a dehydration process without the addition of a secondary crosslinking agent, showed a weight loss rate of over 10% in the dried polymer microparticles before and after high-temperature, high-pressure sterilization (Autoclave) treatment. This indicated that they were not only unsuitable for sterilization and long-term culture, but also showed a swelling degree of 132.43, confirming that the degree of crosslinking was inferior compared to the example. In addition, they exhibited inferior mechanical properties with an average compressive strength of 0.3 mN, confirming that the particles had a low crosslinking density.

[0116] Furthermore, the polymer microparticles of Comparative Example 3, which underwent only primary crosslinking and omitted the secondary crosslinking stage, showed a weight loss rate of over 10% in the dried polymer microparticles before and after high-temperature, high-pressure sterilization (Autoclave) treatment. This indicates that they are not only unsuitable for sterilization and long-term culture, but also showed a swelling degree of 127.79, confirming that the degree of crosslinking was inferior compared to the examples of the present application. In addition, the average compressive strength was 0.23 mN, indicating inferior mechanical properties, confirming that the particles had a low crosslinking density.

Claims

1. A step of forming crosslinked particles by causing a primary crosslinking reaction of a mixture containing an emulsion of a biocompatible polymer and a first crosslinking agent; and A method for producing polymer microparticles, comprising the step of extracting the crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent; The step of forming crosslinked particles by causing a primary crosslinking reaction to a mixture containing the aforementioned emulsion-state biocompatible polymer and a first crosslinking agent is as follows: The step of adding an aqueous solution containing the biocompatible polymer to a hydrophobic solvent to form an emulsion; and The step includes adding a first crosslinking agent to a mixed solution containing the emulsion; The biocompatible polymers mentioned above include hyaluronic acid, carboxymethylcellulose, alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, pullulan, polyglycolide (PGA), polylactide-glycolide copolymer (PLGA), polyanhydride, polyorthoester, polyetherester, polycaprolactone, poly(butyric acid), poly(valeric acid), ethylene-vinyl acetate polymer, and acrylic-substituted cellulose. A method for producing polymer microparticles, wherein the polymer is one or more polymers selected from the group consisting of suacetate, polyvinyl chloride, poly(vinylimidazole), chlorosulfonate polyolefin, polyethylene oxide, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), hydroxypropylcellulose (HPC), cyclodextrin, poly(N-isopropylamide) (PNIPAam), poloxamer, and polyacrylic acid copolymers, and derivatives thereof.

2. In the step of extracting the aforementioned crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent, The method for producing polymer microparticles according to claim 1, wherein the crosslinked particles and the second crosslinking agent undergo a crosslinking reaction in a solid phase.

3. In the step of forming crosslinked particles by subjecting a mixture containing the aforementioned emulsion-state biocompatible polymer and a first crosslinking agent to a primary crosslinking reaction, The method for producing polymer microparticles according to claim 1, wherein the first crosslinking agent is contained in an amount of 30 to 300 parts by weight per 100 parts by weight of the biocompatible polymer.

4. In the step of extracting the aforementioned crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent, The method for producing polymer microparticles according to claim 1, wherein the second crosslinking agent is contained in an amount of 30 to 300 parts by weight per 100 parts by weight of the biocompatible polymer.

5. In the step of extracting the aforementioned crosslinked particles and subjecting them to a secondary crosslinking reaction in an organic solvent phase containing a second crosslinking agent, The method for producing polymer microparticles according to claim 1, wherein the organic solvent containing the second crosslinking agent is an alkaline mixed solvent.

6. The aforementioned organic solvents are ethanol, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylcaprolactam, 2-pyrrolidone, N-ethylpyrrolidone, N-vinylpyrrolidone, dimethyl sulfoxide, tetramethylurea, pyridine, dimethyl sulfone, hexamethyl sulfoxide, gamma-butyrolactone, 3-methoxy-N,N-dimethylpropanamide, 3-ethoxy-N,N-dimethylpropanamide, 3-butoxy-N,N-dimethylpropanamide, 1,3-dimethylimidazolidinone, ethyl amyl ketone, methyl nonyl ketone, methyl ethyl ketone, methyl isoamyl ketone, methyl isopropyl ketone, cyclohexanone, and A method for producing polymer microparticles according to claim 1, wherein the polymer microparticle is one selected from the group consisting of ethylene carbonate, propylene carbonate, diglyme, 4-hydroxy-4-methyl-2-pentanone, ethylene glycol monomethyl ether, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether, ethylene glycol monoethyl ether acetate, ethylene glycol monopropyl ether, ethylene glycol monopropyl ether acetate, ethylene glycol monoisopropyl ether, ethylene glycol monoisopropyl ether acetate, ethylene glycol monobutyl ether, and ethylene glycol monobutyl ether acetate.

7. The method for producing polymer microparticles according to claim 1, wherein the first crosslinking agent and the second crosslinking agent each independently comprise one selected from the group consisting of butanediol diglycidyl ether, ethylene glycol diglycidyl ether, hexanediol diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycol polyglycidyl ether, diglycerol polyglycidyl ether, glycerol polyglycidyl ether, trimethylpropane polyglycidyl ether, 1,2-bis(2,3-epoxypropoxy)ethylene, pentaerythritol polyglycidyl ether, and sorbitol polyglycidyl ether, divinyl sulfone, and glutaraldehyde.

8. It contains a polymer matrix in which biocompatible polymers are crosslinked via a crosslinking agent, The average diameter in distilled water is 1 μm or more and less than 500 μm, The biocompatible polymer is a mixture of hyaluronic acid and gelatin. A first crosslinked region mediated by the first crosslinking agent; and Polymer microparticles containing a second crosslinking region formed by crosslinking a biocompatible polymer via a second crosslinking agent.

9. Polymer microparticles according to claim 8, wherein the degree of swelling according to the following formula 1 is 55 or less: [Math 1]

10. The polymer microparticles according to claim 8, wherein the average compressive strength when deformed to 50% of the average particle diameter is 0.1 mN or more.

11. A medical composition comprising polymer microparticles as described in claim 8 and a pharmaceutically active substance contained within the polymer microparticles.

12. A cosmetic composition comprising polymer microparticles as described in claim 8 and a cosmetic active substance contained within the polymer microparticles.

13. A medical product comprising the medical composition described in claim 11.

14. A beauty product comprising the beauty composition described in claim 12.

Images