WO2017014431A1 - Procédé de préparation de microparticules comprenant un polymère biodégradable - Google Patents
Procédé de préparation de microparticules comprenant un polymère biodégradable Download PDFInfo
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- WO2017014431A1 WO2017014431A1 PCT/KR2016/006209 KR2016006209W WO2017014431A1 WO 2017014431 A1 WO2017014431 A1 WO 2017014431A1 KR 2016006209 W KR2016006209 W KR 2016006209W WO 2017014431 A1 WO2017014431 A1 WO 2017014431A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
- B01J13/06—Making microcapsules or microballoons by phase separation
- B01J13/14—Polymerisation; cross-linking
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
- C08G63/02—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds
- C08G63/06—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds derived from hydroxycarboxylic acids
- C08G63/08—Lactones or lactides
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/12—Powdering or granulating
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L67/00—Compositions of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Compositions of derivatives of such polymers
- C08L67/04—Polyesters derived from hydroxycarboxylic acids, e.g. lactones
Definitions
- the present invention relates to a method for preparing microparticles containing a biodegradable polymer, and more particularly, to a method for preparing microparticles having easy size control, having a constant size distribution, and capable of producing microparticles with high yield.
- Collagen plays a role in maintaining the structure of the tissue, but it does not always maintain a constant shape, but is continuously degraded and reconstituted by an enzyme called collagenase, and reduced by age and photoaging by ultraviolet irradiation.
- This reduction in collagen is known to be closely associated with the formation of wrinkles in the skin (Arthur K. Balin et al., "Aging and the skin", 1989).
- the face part is the most prone to photo-aging in the whole body, and in photo-aged skin, the proliferative activity of fibroblasts, which are the major cells in the dermis, and the synthesis of collagen and the like decrease, and the turnover rate of collagen is also slow. As a result, the skin loses its elasticity, wrinkles occur, and the skin ages.
- products using hyaluronic acid gel has a very good biocompatibility, and has the advantage that there is little biotoxicity as confirmed by various clinical.
- hyaluronic acid has a sustainability problem because reabsorption in vivo occurs very quickly between two weeks and two months.
- products that extend the resorption period in vivo up to 6 months by cross-linking hyaluronic acid and crosslinking material are the mainstream of the market.
- these crosslinked products have also been reported to have problems due to the toxicity of the crosslinking material.
- Sculptra a tissue repair product using biocompatible polymer microparticles that can be degraded in vivo, and this product has polylactic acid as its main raw material. About two years, the effect of tissue repair lasts about two years.
- This product differs in that the hyaluronic acid or collagen product is efficacious due to the hydrated volume of the inserted substance, whereas the inserted polylactic acid forms the patient's own tissue cells and is effective.
- the polylactic acid microspheres or particulates are suspended and injected into carboxymethylcellulose, wherein the microspheres or particulates must be at least 20 ⁇ m in diameter so as not to be attracted to the macrophage phagocytes.
- this product does not show efficacy immediately after infusion and shows efficacy after about 3 months.
- the carboxymethyl cellulose used as a suspending agent takes a long time to hydrate and dissolve in water has a disadvantage that must be dissolved in water before 2 to 24 hours before the procedure, clogging the syringe needle due to the particle size of the microspheres It also has a lot of problems. Thus, without causing any unwanted reactions in the human body, it is possible to avoid aggregation, needle clogging and nodule forming at the time of infusion, and due to spherical microspheres There is a need to develop slow resorbable biodegradable fillers such as flow characteristics.
- Korean Patent No. 10-1517258 relates to a method for preparing microparticles using a solvent extraction-evaporation method including polycaprolactone, and a method for preparing microparticles using the solvent extraction-evaporation method includes:
- the disadvantage is that the size is not uniform and the size distribution is relatively wide. Therefore, in order to obtain the microparticles of the desired size, a separate size purification process is required, and since the microparticles of unnecessary size are also produced, there is a problem in that the final yield falls.
- An object of the present invention is to provide a method for producing microparticles containing a biodegradable polymer.
- the present invention can control the size according to the water injection rate, the water injection angle, the biodegradable polymer solution injection rate and the biodegradable polymer solution concentration, the method of producing a microparticle containing a biodegradable polymer having a constant size distribution. To provide another purpose.
- Another object of the present invention is to provide a spherical microparticle having a spherical microparticle for injection into the skin.
- the present invention comprises the steps of (1) dissolving the biodegradable polymer in an organic solvent to prepare a biodegradable polymer solution; (2) dissolving the surfactant in purified water to prepare an aqueous solution; (3) injecting the biodegradable polymer solution of step (1) in one direction to allow flow; (4) injecting the aqueous phase solution in the flow direction of step (3) to allow the first and second streams to flow, and the intersection of the flow of step (3), the first flow of the aqueous phase solution, and the second flow.
- step (8) relates to a method for producing a microparticle comprising a biodegradable polymer comprising the step of secondary drying the spherical microparticles of step (7) by vacuum or freeze drying.
- the manufacturing method can be largely divided into three flow stems.
- the first flow stem is a flow stem of a biodegradable polymer solution prepared by dissolving the biodegradable polymer in an organic solvent.
- the second flow stem is the flow stem of the aqueous solution prepared by dissolving the surfactant in purified water.
- the third flow stem is a flow stem generated as one flow after the first flow stem and the second flow stem create an intersection point.
- the first flow stem and the third flow stem flow in the same direction, and the second flow stem is more specifically distinguished into the first flow and the second flow of the aqueous phase solution, and flows in the direction of creating an intersection with the first flow.
- the flow rate of the three flow stems and the injection angle of the second flow will affect the size, size distribution and production yield of the resulting microparticles.
- the relationship between the aqueous solution injection rate (mL), the biodegradable polymer solution injection rate (mL) and the biodegradable polymer solution concentration of the present invention is represented by the following equation 1, the following equation 1 is 2.5 It relates to a method for producing a microparticle having a value of from 15 to 15.
- Equation 1 water phase injection rate (mL)
- OF biodegradable polymer solution injection rate (mL)
- OC biodegradable polymer solution concentration (% by weight)
- K proportional constant (proportional factor). If the value of Equation 1 is less than 2.5 or more than 15, the size and size of the microparticles may be due to the low shear stress due to the change of the injection rate of the biodegradable polymer solution, the difference in the shear force applied to the biodegradable polymer solution, and the like. The distribution increased.
- the aqueous solution of step (2) of the present invention has a surfactant concentration of 0.25 to 0.5% by weight.
- Increasing the surfactant concentration of the aqueous phase solution can suppress the aggregation of microparticles in the manufacturing process, and the low interfacial tension produces microparticles with a narrow size distribution.
- the surfactant concentration of the aqueous solution is less than 0.25% by weight, there is a problem that the size distribution of the microparticles is widened.
- a problem of relatively difficult washing and removing of the surfactant in the generated microparticles may occur.
- the biodegradable polymer solution of step (3) of the present invention is to flow at a rate of 50 to 200ul / min.
- the biodegradable polymer solution is allowed to flow at a rate of less than 50 ⁇ l / min, the size of the microparticles may be small, so that they may be absorbed into the body, and the size distribution may be widened. If the flow exceeds 200 ⁇ l / min, the size of the microparticles increases, so that a wound may occur when injected into the skin, and there is a problem that the size distribution is widened.
- the biodegradable polymer solution of step (3) of the present invention has a concentration of 5 to 30% by weight.
- concentration of the biodegradable polymer solution is less than 5% by weight, the concentration of the biodegradable polymer is too low, there is a problem that the production of microparticles is difficult, and when it exceeds 30% by weight there is a problem that it is difficult to produce spherical microparticles .
- the first flow and the second flow of the aqueous phase solution of step (4) is a flow of 30 to 90 degrees to both sides based on the flow of the step (3).
- the flow of the biodegradable polymer solution in step (3) is a flow having a direction from the starting point (a) to the injection into the biodegradable polymer solution to a certain point (b),
- the flow of the solution is a flow to generate the intersection (d) and the flow of the biodegradable polymer solution from the starting point (c, c ') to be injected and flowed.
- the aqueous phase solution of step (4) is to flow at a rate of 100 to 2000ul / min.
- the first and second streams of the aqueous phase solution are allowed to flow at a rate of 100 to 2000 ⁇ l / min, preferably at a rate of 500 to 1000 ⁇ l / min. If the rate of the aqueous solution is less than 500 ⁇ l / min, a problem arises in that the size range of the microparticles is widened. If so, there is a problem that the biodegradation period is too short because it is rapidly decomposed.
- the relatively slow biodegradable polymer solution at the intersection of the biodegradable polymer solution stream and the aqueous phase solution stream is squeezed by the relatively high aqueous phase solution on both sides of the device, so that a small amount of the biodegradable polymer solution is disconnected and surrounded by the aqueous phase solution. do.
- the small amount of biodegradable polymer solution immediately takes the form of spherical microparticles due to the surface tension.
- the step of collecting the spherical microparticles produced in the single stream after the intersection and the intersection of step (5) of the present invention is collected in a receiver solution containing a surfactant.
- the capture in the receiver solution containing the surfactant is to prevent the spherical microparticles from agglomerating with each other.
- step (5) collects spherical microparticles in a negative pressure environment. Negative pressure environment means maintaining a pressure lower than atmospheric pressure. Therefore, when the microparticles are collected in a negative pressure environment, the low pressure acts to pull the flow of the biodegradable polymer solution and the water phase solution, which causes the pump to flow the biodegradable polymer solution and the aqueous solution.
- step (6) of the present invention is to first dry the spherical microparticles collected in step (5) at 20 to 25 °C. If the emulsion in droplet form is held at a temperature below the boiling point of the organic solvent for a period of time, for example 12 to 48 hours, the organic solvent can be extracted from the biodegradable polymer solution in the form of droplets. As the organic solvent is extracted from the biodegradable polymer solution in the form of droplets, it can be solidified to form spherical microparticles.
- step (7) of the present invention is to filter and wash the dried spherical microparticles of step (6), the microparticles in purified water at least once, preferably 1 to 3 times Washing can remove the surfactant and solvent and again filter to obtain washed particulates.
- the washing step to remove remaining surfactant and solvent can be repeated several times.
- step (8) of the present invention is to secondary dry the spherical microparticles of step (7), in the step (8), the drying method is not particularly limited, but preferably In order to minimize the damage of the biodegradable polymer by heat, it may be carried out through vacuum drying or freeze drying.
- the average diameter of the microparticles of the present invention is 10 to 150 mu m, preferably 15 to 90 mu m, more preferably 20 to 70 mu m.
- Micro particles are spherical micro particles, when the diameter is less than 10 ⁇ m injected into the body, there is a problem that the biodegradation period is too short to form enough tissue cells because the biodegradation is too short. If the diameter exceeds 150 ⁇ m, there is a problem that the injection into the skin is not easy.
- the microparticles of the present invention is a method for producing microparticles comprising a biodegradable polymer having a size distribution of 1 or less (span value) according to Equation 2 below:
- the biodegradable polymer of the present invention is polylactic acid (Polylactic acid, PLA), polyglycolic acid (PGA), polylactic acid-glycolic acid copolymer (Poly (lactic-co-glycolic acid) ), PLGA), polycaprolactone (PCL) and derivatives thereof, and at least one selected from the group consisting of polycaprolactone, preferably polycaprolactone, but not limited to the examples.
- the number average molecular weight of the biodegradable polymer is not particularly limited, but is 5,000 to 300,000, preferably 8,000 to 250,000, and more preferably 10,000 to 200,000.
- the organic solvent of the present invention has a boiling point of 120 ° C. or less and is not mixed with water, for example, with dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane and mixtures thereof. At least one selected from the group consisting of, preferably dichloromethane, but is not limited to the examples.
- the surfactant is not particularly limited in kind, and may be used as long as the biodegradable polymer solution can help to form a stable emulsion.
- the method for producing microparticles containing the biodegradable polymer of the present invention can be used to apply the microfluidic method. That is, a microchannel through which the biodegradable polymer solution and the aqueous solution may flow is made in the device, and the biodegradable polymer solution and the aqueous solution are injected through the microchannel to flow.
- the channel is formed using a deep ion reactive etching (DRIE) process on a silicon wafer to etch in a vertical direction to form a valley, and then anodically bond and seal the glass thereon.
- the microchannel is prepared to have a width of 300 ⁇ m and a depth of 150 ⁇ m to 200 ⁇ m. The reason why the DRIE is used in the manufacturing process is that when performing the etching of 50 ⁇ m or more as described above, it is possible to etch the surface shape vertically differently from other wet etching.
- Preparation of microparticles using a solvent extraction-evaporation method as in the prior art has disadvantages in that the size of the microparticles is not uniform and the size distribution is relatively wide. Therefore, in order to obtain the microparticles of the desired size, a separate size purification process is required, and since the microparticles of unnecessary size are also produced, there is a problem in that the final yield falls.
- the phase separation property of the biodegradable polymer solution and the surfactant solution is the intersection and cross point of the biodegradable polymer solution and the aqueous solution Implemented in a single flow thereafter, it is easier to control the size than the conventional solvent extraction-evaporation method, has a high final production yield, and has excellent mechanical strength and shape of microparticles.
- it is easy to control the degree of biodegradation, less penetrability and foreign body feeling.
- the microparticles containing the biodegradable polymer prepared by the production method of the present invention is not particularly limited in use thereof, for example, a skin cosmetic or medical filler that requires in vivo resorption It can be used as, but is not limited to, examples, especially as an injectable subcutaneous or intradermal injectable filler.
- the in vivo reabsorption period of the microparticles containing the biodegradable polymer prepared according to the preparation method of the present invention is not particularly limited, but may be considered to be used as a biodegradable skin cosmetic or medical filler. At this time, it is preferable that the bioabsorbable can be revived within 1 to 3 years.
- the present invention is a method for producing microparticles containing a biodegradable polymer, specifically a method for producing spherical microparticles for injection into the skin. Due to the manufacturing method described above, it is possible to control the size of the microparticles, and to produce microparticles having a constant size distribution. As microparticles containing biodegradable polymers can be produced to have a narrow size range, high production yields can be exhibited. In addition, in consideration of the characteristic that the decomposition rate varies depending on the size of the microparticles, the rate of decomposition of the microparticles in the body can be controlled by adjusting the size of the microparticles to be injected as necessary.
- the molecular weight of the polymer means number average molecular weight (Mn).
- Mn number average molecular weight
- the number average molecular weight means an average molecular weight obtained by averaging the molecular weights of the component molecular species of the polymer compound having a molecular weight distribution by a moisture content or a mole fraction.
- the tissue repair refers to the mechanism of restoring the tissue to its original state when necrosis or defect occurs in the tissue due to trauma or inflammation.
- the biodegradable polymer means a property that the medical material is harmless and easy to adapt to a living body.
- the living organisms are broadly divided into blood, various tissues and organs, and thus are divided into blood compatibility (antithrombogenic) and tissue compatibility.
- the biodegradable polymer means a polymer that can be widely applied.
- the present invention comprises the steps of (1) dissolving the biodegradable polymer in an organic solvent to prepare a biodegradable polymer solution; (2) dissolving the surfactant in purified water to prepare an aqueous solution; (3) injecting the biodegradable polymer solution of step (1) in one direction to allow flow; (4) injecting the aqueous phase solution in the flow direction of step (3) to allow the first and second streams to flow, and the intersection of the flow of step (3), the first flow of the aqueous phase solution, and the second flow.
- step (8) relates to a method for producing a microparticle comprising a biodegradable polymer comprising the step of secondary drying the spherical microparticles of step (7) by vacuum or freeze drying.
- the manufacturing method can be largely divided into three flow stems.
- the first flow stem is a flow stem of a biodegradable polymer solution prepared by dissolving the biodegradable polymer in an organic solvent.
- the second flow stem is the flow stem of the aqueous solution prepared by dissolving the surfactant in purified water.
- the third flow stem is a flow stem generated as one flow after the first flow stem and the second flow stem create an intersection point.
- the first flow stem and the third flow stem flow in the same direction, and the second flow stem is more specifically distinguished into the first flow and the second flow of the aqueous phase solution, and flows in the direction of creating an intersection with the first flow.
- the flow rate of the three flow stems and the injection angle of the second flow will affect the size, size distribution and production yield of the resulting microparticles.
- the relationship between the aqueous solution injection rate (mL), the biodegradable polymer solution injection rate (mL) and the biodegradable polymer solution concentration of the present invention is represented by the following equation 1, the following equation 1 is 2.5 It relates to a method for producing a microparticle having a value of from 15 to 15.
- Equation 1 water phase injection rate (mL)
- OF biodegradable polymer solution injection rate (mL)
- OC biodegradable polymer solution concentration (% by weight)
- K proportional constant (proportional factor). If the value of Equation 1 is less than 2.5 or more than 15, the size and size of the microparticles may be due to the low shear stress due to the change of the injection rate of the biodegradable polymer solution, the difference in the shear force applied to the biodegradable polymer solution, and the like. The distribution increased.
- the aqueous solution of step (2) of the present invention has a surfactant concentration of 0.25 to 0.5% by weight.
- Increasing the surfactant concentration of the aqueous phase solution can suppress the aggregation of microparticles in the manufacturing process, and the low interfacial tension produces microparticles with a narrow size distribution.
- the surfactant concentration of the aqueous solution is less than 0.25% by weight, there is a problem that the size distribution of the microparticles is widened.
- a problem of relatively difficult washing and removing of the surfactant in the generated microparticles may occur.
- the biodegradable polymer solution of step (3) of the present invention is to flow at a rate of 50 to 200ul / min.
- the biodegradable polymer solution is allowed to flow at a rate of less than 50 ⁇ l / min, the size of the microparticles may be small, so that they may be absorbed into the body, and the size distribution may be widened. If the flow exceeds 200 ⁇ l / min, the size of the microparticles increases, so that a wound may occur when injected into the skin, and there is a problem that the size distribution is widened.
- the biodegradable polymer solution of step (3) of the present invention has a concentration of 5 to 30% by weight.
- concentration of the biodegradable polymer solution is less than 5% by weight, the concentration of the biodegradable polymer is too low, there is a problem that the production of microparticles is difficult, and when it exceeds 30% by weight there is a problem that it is difficult to produce spherical microparticles .
- the first flow and the second flow of the aqueous phase solution of step (4) is a flow of 30 to 90 degrees to both sides based on the flow of the step (3).
- the flow of the biodegradable polymer solution in step (3) is a flow having a direction from the starting point (a) to the injection into the biodegradable polymer solution to a certain point (b),
- the flow of the solution is a flow to generate the intersection (d) and the flow of the biodegradable polymer solution from the starting point (c, c ') to be injected and flowed.
- the aqueous phase solution of step (4) is to flow at a rate of 100 to 2000ul / min.
- the first and second streams of the aqueous phase solution are allowed to flow at a rate of 100 to 2000 ⁇ l / min, preferably at a rate of 500 to 1000 ⁇ l / min. If the rate of the aqueous solution is less than 500 ⁇ l / min, a problem arises in that the size range of the microparticles is widened. If so, there is a problem that the biodegradation period is too short because it is rapidly decomposed.
- the relatively slow biodegradable polymer solution at the intersection of the biodegradable polymer solution stream and the aqueous phase solution stream is squeezed by the relatively high aqueous phase solution on both sides of the device, so that a small amount of the biodegradable polymer solution is disconnected and surrounded by the aqueous phase solution. do.
- the small amount of biodegradable polymer solution immediately takes the form of spherical microparticles due to the surface tension.
- the step of collecting the spherical microparticles produced in a single stream after the intersection and the intersection of step (5) of the present invention is collected in a receiver solution containing a surfactant.
- the capture in the receiver solution containing the surfactant is to prevent the spherical microparticles from agglomerating with each other.
- step (5) collects spherical microparticles in a negative pressure environment. Negative pressure environment means maintaining a pressure lower than atmospheric pressure. Therefore, when the microparticles are collected in a negative pressure environment, the low pressure acts to pull the flow of the biodegradable polymer solution and the water phase solution, which causes the pump to flow the biodegradable polymer solution and the aqueous solution.
- step (6) of the present invention is to first dry the spherical microparticles collected in step (5) at 20 to 25 °C. If the emulsion in droplet form is maintained at a temperature below the boiling point of the organic solvent for a period of time, for example 12 to 48 hours, the organic solvent can be extracted from the biodegradable polymer solution in the form of droplets. As the organic solvent is extracted from the biodegradable polymer solution in the form of droplets, it can be solidified to form spherical microparticles.
- step (7) of the present invention is to filter and wash the dried spherical microparticles of step (6), the microparticles in purified water at least once, preferably 1 to 3 times Washing can remove the surfactant and solvent and again filter to obtain washed particulates.
- the washing step to remove remaining surfactant and solvent can be repeated several times.
- step (8) of the present invention is to secondary dry the spherical microparticles of step (7), in the step (8), the drying method is not particularly limited, but preferably In order to minimize the damage of the biodegradable polymer by heat, it may be carried out through vacuum drying or freeze drying.
- the average diameter of the microparticles of the present invention is 10 to 150 mu m, preferably 15 to 90 mu m, more preferably 20 to 70 mu m.
- Micro particles are spherical micro particles, when the diameter is less than 10 ⁇ m injected into the body, there is a problem that the biodegradation period is too short to form enough tissue cells because the biodegradation is too short. If the diameter exceeds 150 ⁇ m, there is a problem that the injection into the skin is not easy.
- the microparticles of the present invention is a method for producing microparticles comprising a biodegradable polymer having a size distribution of 1 or less (span value) according to Equation 2 below:
- the biodegradable polymer of the present invention is polylactic acid (Polylactic acid, PLA), polyglycolic acid (PGA), polylactic acid-glycolic acid copolymer (Poly (lactic-co-glycolic acid) ), PLGA), polycaprolactone (PCL) and derivatives thereof, and at least one selected from the group consisting of polycaprolactone, preferably polycaprolactone, but not limited to the examples.
- the number average molecular weight of the biodegradable polymer is not particularly limited, but is 5,000 to 300,000, preferably 8,000 to 250,000, and more preferably 10,000 to 200,000.
- the organic solvent of the present invention has a boiling point of 120 ° C. or less and is not mixed with water, for example, with dichloromethane, chloroform, chloroethane, dichloroethane, trichloroethane and mixtures thereof. At least one selected from the group consisting of, preferably dichloromethane, but is not limited to the examples.
- the surfactant is not particularly limited in kind, and may be used as long as the biodegradable polymer solution can help to form a stable emulsion.
- the method for producing microparticles containing the biodegradable polymer of the present invention can be used to apply the microfluidic method. That is, a microchannel through which the biodegradable polymer solution and the aqueous solution may flow is made in the device, and the biodegradable polymer solution and the aqueous solution are injected through the microchannel to flow.
- the channel is formed using a deep ion reactive etching (DRIE) process on a silicon wafer to etch in a vertical direction to form a valley, and then anodically bond and seal the glass thereon.
- the microchannel is prepared to have a width of 300 ⁇ m and a depth of 150 ⁇ m to 200 ⁇ m. The reason why the DRIE is used in the manufacturing process is that when performing the etching of 50 ⁇ m or more as described above, it is possible to etch the surface shape vertically differently from other wet etching.
- Preparation of microparticles using a solvent extraction-evaporation method as in the prior art has disadvantages in that the size of the microparticles is not uniform and the size distribution is relatively wide. Therefore, in order to obtain the microparticles of the desired size, a separate size purification process is required, and since the microparticles of unnecessary size are also produced, there is a problem in that the final yield falls.
- the phase separation property of the biodegradable polymer solution and the surfactant solution is the intersection and cross point of the biodegradable polymer solution and the aqueous solution Implemented in a single flow thereafter, it is easier to control the size than the conventional solvent extraction-evaporation method, has a high final production yield, and has excellent mechanical strength and shape of microparticles.
- it is easy to control the degree of biodegradation, less penetrability and foreign body feeling.
- the microparticles containing the biodegradable polymer prepared by the production method of the present invention is not particularly limited in use thereof, for example, a skin cosmetic or medical filler that requires in vivo resorption It can be used as, but is not limited to, examples, especially as an injectable subcutaneous or intradermal injectable filler.
- the in vivo reabsorption period of the microparticles containing the biodegradable polymer prepared according to the preparation method of the present invention is not particularly limited, but may be considered to be used as a biodegradable skin cosmetic or medical filler. At this time, it is preferable that the bioabsorbable can be revived within 1 to 3 years.
- a biodegradable polymer solution 10 mL of a biodegradable polymer solution was prepared by dissolving polycaprolactone (PCL), Mn 45,000, in 15 wt% in a solvent of dichloromethane (boiling point: 39.6 ° C.).
- PCL polycaprolactone
- Mn 45,000 molecular weight of 85,000 to 124,000 was dissolved in purified water at 0.25% by weight to prepare 250 mL of an aqueous solution.
- PVA polyvinyl alcohol
- a dispersed phase in the form of droplets was formed.
- the dispersed phase in the form of droplets was dissolved in purified water at 0.25% by weight of polyvinyl alcohol (PVA), collected in 100 mL of a receiver solution, and left at room temperature (25 ° C.) for about 24 hours to extract a dichloromethane solvent.
- PVA polyvinyl alcohol
- the aqueous solution containing microparticles is filtered and the microparticles are washed to remove residual polyvinyl alcohol and dichloromethane solution. Thereafter, the microparticles were dried to finally prepare a microparticle including a spherical biodegradable polymer.
- Polycaprolactone (Polycaprolactone, PCL) of Mn 45,000 was prepared in the same manner as in Preparation Example 1, except that 10 mL of a biodegradable polymer solution was prepared by dissolving 10 wt% in a dichloromethane (boiling point: 39.6 ° C.) solvent.
- Polycaprolactone (Polycaprolactone, PCL) of Mn 45,000 was prepared in the same manner as in Preparation Example 1, except that 10 mL of a biodegradable polymer solution was prepared by dissolving in a solvent of dichloromethane (boiling point: 39.6 ° C.) at 30% by weight.
- Biodegradable polymer solution was prepared in the same manner as in Preparation Example 1 except that the flow rate was 50 ⁇ l / min.
- Biodegradable polymer solution was prepared in the same manner as in Preparation Example 1 except that the flow at a rate of 200 ⁇ l / min.
- the preparation was carried out in the same manner as in Preparation Example 1, except that the surfactant concentration of the aqueous phase solution was dissolved at 0.5% by weight.
- Polycaprolactone (Polycaprolactone, PCL) of Mn 45,000 was prepared in the same manner as in Preparation Example 1 except that 10 mL of a biodegradable polymer solution was prepared by dissolving 5 wt% in a solvent of dichloromethane (boiling point: 39.6 ° C.).
- Polycaprolactone (Polycaprolactone, PCL) of Mn 45,000 was prepared in the same manner as in Preparation Example 1, except that 10 mL of a biodegradable polymer solution was prepared by dissolving 45 wt% in a solvent of dichloromethane (boiling point: 39.6 ° C).
- Biodegradable polymer solution was prepared in the same manner as in Preparation Example 1 except that the flow rate was 25 ⁇ l / min.
- Biodegradable polymer solution was prepared in the same manner as in Preparation Example 1 except that the flow at a rate of 300 ⁇ l / min.
- the preparation was carried out in the same manner as in Preparation Example 1, except that the surfactant concentration of the aqueous solution was dissolved at 0.1 wt%.
- the preparation was carried out in the same manner as in Preparation Example 1, except that the surfactant concentration of the aqueous solution was prepared by dissolving at 1.0 wt%.
- the biodegradable polymer solution was prepared in the same manner as in Preparation Example 3 except that the flow was flowed at a rate of 200 ⁇ l / min, and the first and second flows of the aqueous phase solution were flowed at a rate of 2000 ⁇ l / min.
- the biodegradable polymer solution was prepared in the same manner as in Preparation Example 3 except that the flow was flowed at a rate of 50 ⁇ l / min, and the first and second flows of the aqueous phase solution were flowed at a rate of 2000 ⁇ l / min.
- a biodegradable polymer solution 10 mL of a biodegradable polymer solution was prepared by dissolving polycaprolactone (PCL), Mn 45,000, in 15 wt% in a solvent of dichloromethane (boiling point: 39.6 ° C.).
- PCL polycaprolactone
- Mn 45,000 10 wt%
- a solvent of dichloromethane solvent of dichloromethane
- Mw 85,000-124,000 polyvinyl alcohol (PVA) was dissolved as 0.25% wt in purified water to prepare 100 mL of an aqueous solution.
- the biodegradable polymer solution was then mixed with the surfactant aqueous phase solution and then stirred rapidly to form a dispersed phase in the form of droplets.
- the dispersed phase in the form of droplets dispersed in the surfactant aqueous phase solution was then stirred for a period of time and the temperature was added to extract the solvent and solidified to form microparticles. Thereafter, the solidified microparticles were filtered and washed with purified water to remove residual surfactant. Subsequently, the washed microparticles are obtained, followed by a drying process to finally form spherical biodegradable microparticles.
- the size and distribution of the microparticles containing the biodegradable polymer were measured by repeating three times for 10 seconds after dispersing the microparticles in a dispersion medium using a Helos particle size analyzer (Sympatec) as a measuring device.
- a Helos particle size analyzer Sympatec
- Biodegradable Polymer Molecular Weight Mn
- Biodegradable polymer solution concentration % by weight
- Biodegradation Polymer Solution Injection Rate ⁇ l / min
- Water phase injection rate ⁇ l / min
- Microparticle size D10 / D50 / D90, um
- Span value Preparation Example 1 45,000 15 100 1,000 25/40/51, 0.65 Comparative Example 1 14,000 15 100 1,000 32/48/75, 0.90 Comparative Example 2 80,000 15 100 1,000 23/31/42, 0.61
- polycaprolactone (Polycaprolactone, PCL) microparticles having a molecular weight of Mn 45,000 were confirmed to have a uniform size and a perfect spherical shape, the bioabsorption period to be developed in the present invention could be secured.
- polycaprolactone (PCL) microparticles having a molecular weight of Mn 14,000 have a wide size distribution and do not show perfect spherical shape due to low molecular weight.
- PCL polycaprolactone
- Biodegradable Polymer Molecular Weight Mn
- Biodegradable polymer solution concentration % by weight
- Biodegradation Polymer Solution Injection Rate ⁇ l / min
- Water phase injection rate ⁇ l / min
- Microparticle size D10 / D50 / D90, um
- Span value Optimization Constant OC
- the microparticles having a concentration of 10 to 30% by weight of the biodegradable polymer solution in Preparation Examples 1 to 3 were uniform in size and were able to confirm the perfect spherical shape.
- the concentration of the biodegradable polymer solution in Comparative Example 3 is 5% by weight, very small sized microparticles are generated due to the low concentration, and the produced microparticles are porous and do not meet the purpose of this study.
- Comparative Example 4 when the concentration of the biodegradable polymer solution was 45% by weight, it was difficult to form spherical microparticles due to the high concentration.
- Biodegradable Polymer Molecular Weight Mn
- Biodegradable polymer solution concentration % by weight
- Biodegradation Polymer Solution Injection Rate ⁇ l / min
- Water phase injection rate ⁇ l / min
- Microparticle size D10 / D50 / D90, um
- Span value Optimization constant OC
- the microparticles of the injection rate of the biodegradable polymer solution in Preparation Examples 1, 4, 5 is 50 to 200 ⁇ l / min is a perfect sphere of the desired size (20 ⁇ 70 ⁇ m) intended in this study I could confirm the form.
- the injection rate of the biodegradable polymer solution in Comparative Example 5 is 25 ⁇ l / min, small microparticles are generated, but the size distribution of the microparticles is increased.
- Comparative Example 6 when the injection rate of the biodegradable polymer solution was 300 ⁇ l / min, the size and size distribution of the microparticles were increased due to the low shear stress.
- Biodegradable polymer solution concentration (% by weight) Biodegradation Polymer Solution Injection Rate ( ⁇ l / min) Water phase injection rate ( ⁇ l / min) Water solution injection angle Microparticle size (D10 / D50 / D90, um) and Span value Optimization constant (OC)
- Preparation Example 1 15 100 1,000 90 25/40/51, 0.65 6.1
- Preparation Example 6 15 100 1,000 60 28/44/59, 0.70 2.7
- Comparative Example 7 15 100 1,000 30 34/43/74, 0.93 0.7 Comparative Example 8 15 100 1,000 150 29/44/71, 0.95 16.9
- Biodegradable polymer solution concentration (% by weight) Biodegradation Polymer Solution Injection Rate ( ⁇ l / min) Water phase injection rate ( ⁇ l / min) Water solution injection angle Microparticle size (D10 / D50 / D90, um) and Span value Optimization constant (OC)
- Preparation Example 1 15 100 1,000 90 25/40/51, 0.65 6.1
- Preparation Example 7 15 100 750 90 27/42/63, 0.86 3.4
- Comparative Example 9 15 100 500 90 34/52/83, 0.94 1.5 Comparative Example 10 15 100 2,000 90 16/29/49, 1.14 24.3
- microparticles having an injection rate of 750 to 1000 ⁇ l / min were confirmed to have a uniform size and a perfect spherical shape.
- the injection rate of the aqueous phase solution in Comparative Example 9 is 500 ⁇ l / min because the shear force applied to the polymer solution is insufficient to form a spherical microparticles, the size of the microparticles are large and does not take the perfect spherical shape.
- the injection rate of the aqueous phase solution in Comparative Example 10 is 2000 ⁇ l / min, small microparticles are generated due to high shear force, but the size distribution was confirmed to be increased.
- Biodegradable polymer solution concentration (% by weight) Biodegradation Polymer Solution Injection Rate ( ⁇ l / min) Water phase injection rate ( ⁇ l / min) Aqueous solution concentration (% by weight) Microparticle size (D10 / D50 / D90, um) and Span value Optimization constant (OC)
- Preparation Example 1 15 100 1,000 0.25 25/40/51, 0.65 6.1
- Preparation Example 8 15 100 1,000 0.5 22/36/46, 0.67 12.2
- Comparative Example 11 15 100 1,000 0.1 29/44/71, 0.95 2.4
- Comparative Example 12 15 100 1,000 One 21/33/41, 0.61 24.3
- the microparticles having a concentration of 0.25 to 0.5% by weight of the aqueous solution could be confirmed to have a uniform size and a perfect spherical shape.
- the concentration of the aqueous solution in Comparative Example 11 is 0.1% by weight
- the surface tension of the microparticles is increased due to the low surfactant concentration, which results in an increase in the size of the generated microparticles.
- the concentration of the aqueous phase solution in Comparative Example 12 is 1.0% by weight, it is possible to suppress the agglomeration of the microparticles during the manufacturing process, and microparticles having a narrow size distribution are generated due to the low interfacial tension.
- the PVA concentration of the aqueous phase solution is 1.0% wt or more, it is difficult to wash and remove PVA from the produced microparticles.
- Biodegradable Polymer Molecular Weight Mn
- Biodegradable polymer solution concentration % by weight
- Biodegradation Polymer Solution Injection Rate ⁇ l / min
- Water phase injection rate ⁇ l / min
- Microparticle size D10 / D50 / D90, um
- Span value Optimization constant OC
- a biodegradable polymer solution was prepared and injected using a device having a microchannel, and the aqueous phase solution was injected at an angle of 90 ° to form microparticles, followed by filtration and Microparticles obtained through washing were prepared by drying.
- the microparticles were prepared from the biodegradable polymer solution of the same concentration using polycaprolactone (PCL) having the same molecular weight as that of Preparation Example 1 by a solvent extraction-evaporation method, which is a general method for preparing microparticles. It was.
- PCL polycaprolactone
- Comparative Example 16 prepared by the solvent extraction-evaporation method unlike the Preparation Example 1, the size of the microparticles was large, and it was confirmed that the size distribution was relatively wide.
- the present invention relates to a method for preparing microparticles containing a biodegradable polymer, and more particularly, to a method for preparing microparticles having easy size control, having a constant size distribution, and capable of producing microparticles with high yield.
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Abstract
La présente invention porte sur un procédé de préparation de microparticules comprenant un polymère biodégradable tel que les microparticules préparées selon la présente invention sont des microparticules sphériques pour l'injection cutanée. La taille des microparticules peut être contrôlée au moyen du procédé de préparation. De plus, les microparticules comprenant un polymère biodégradable peuvent être produites dans une plage de tailles étroite et peuvent ainsi présenter un rendement de production élevé. Considérant la caractéristique dans laquelle un taux de dissolution varie en fonction de la taille des microparticules, le taux de dissolution peut être contrôlé après l'injection des microparticules, préparées selon la présente invention, dans un corps.
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KR10-2015-0101511 | 2015-07-17 | ||
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KR1020160013221A KR101726559B1 (ko) | 2015-07-17 | 2016-02-03 | 생분해성 고분자를 포함하는 마이크로파티클의 제조방법 |
KR10-2016-0013221 | 2016-02-03 |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110681323A (zh) * | 2019-08-26 | 2020-01-14 | 上海摩漾生物科技有限公司 | 具有微拓扑结构的高尔夫球型可降解微球及其制备方法 |
EP3570970A4 (fr) * | 2016-11-14 | 2021-05-05 | Inventage Lab. Inc. | Optimisation de la conception d'appareils et procédés pour la production en masse de microsphères à base de polymère biodégradable monodispersés et de systèmes d'administration de médicament à base de polymère biodégradable |
CN114146020A (zh) * | 2022-02-10 | 2022-03-08 | 中国远大集团有限责任公司 | 一种注射美容产品及其制备方法和应用 |
US20230071710A1 (en) * | 2020-02-03 | 2023-03-09 | Inventage Lab Inc. | Microparticle producing system which comprises carrying fluid, and a controlling method thereof |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009014441A2 (fr) * | 2007-07-26 | 2009-01-29 | Aqtis Ip Bv | Microparticules comprenant plc et leurs utilisations |
KR20110121320A (ko) * | 2010-04-30 | 2011-11-07 | 삼일제약주식회사 | 나노입자, 나노입자의 제조 방법 및 제조 장치 |
JP2012011268A (ja) * | 2010-06-29 | 2012-01-19 | Okayama Univ | 生分解性中空微粒子およびその製造方法 |
KR20140013533A (ko) * | 2012-07-25 | 2014-02-05 | 황재광 | 화장용 브러쉬 제조장치 및 그 제조방법 |
KR20140128553A (ko) * | 2013-04-26 | 2014-11-06 | 공주대학교 산학협력단 | 활성물질을 담지할 수 있는 표면 딤플구조를 갖는 미립자 및 그 제조방법 |
-
2016
- 2016-06-10 WO PCT/KR2016/006209 patent/WO2017014431A1/fr active Application Filing
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009014441A2 (fr) * | 2007-07-26 | 2009-01-29 | Aqtis Ip Bv | Microparticules comprenant plc et leurs utilisations |
KR20110121320A (ko) * | 2010-04-30 | 2011-11-07 | 삼일제약주식회사 | 나노입자, 나노입자의 제조 방법 및 제조 장치 |
JP2012011268A (ja) * | 2010-06-29 | 2012-01-19 | Okayama Univ | 生分解性中空微粒子およびその製造方法 |
KR20140013533A (ko) * | 2012-07-25 | 2014-02-05 | 황재광 | 화장용 브러쉬 제조장치 및 그 제조방법 |
KR20140128553A (ko) * | 2013-04-26 | 2014-11-06 | 공주대학교 산학협력단 | 활성물질을 담지할 수 있는 표면 딤플구조를 갖는 미립자 및 그 제조방법 |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3570970A4 (fr) * | 2016-11-14 | 2021-05-05 | Inventage Lab. Inc. | Optimisation de la conception d'appareils et procédés pour la production en masse de microsphères à base de polymère biodégradable monodispersés et de systèmes d'administration de médicament à base de polymère biodégradable |
CN110681323A (zh) * | 2019-08-26 | 2020-01-14 | 上海摩漾生物科技有限公司 | 具有微拓扑结构的高尔夫球型可降解微球及其制备方法 |
CN110681323B (zh) * | 2019-08-26 | 2021-12-21 | 上海摩漾生物科技有限公司 | 具有微拓扑结构的高尔夫球型可降解微球及其制备方法 |
US20230071710A1 (en) * | 2020-02-03 | 2023-03-09 | Inventage Lab Inc. | Microparticle producing system which comprises carrying fluid, and a controlling method thereof |
US11731100B2 (en) * | 2020-02-03 | 2023-08-22 | Inventage Lab Inc. | Microparticle producing system which comprises carrying fluid, and a controlling method thereof |
CN114146020A (zh) * | 2022-02-10 | 2022-03-08 | 中国远大集团有限责任公司 | 一种注射美容产品及其制备方法和应用 |
CN114146020B (zh) * | 2022-02-10 | 2022-04-29 | 中国远大集团有限责任公司 | 一种注射美容产品及其制备方法和应用 |
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