Filler composition as well as preparation method and application thereof

Abstract

The invention discloses a filler composition as well as a preparation method and application thereof. The preparation method of the filler composition comprises the following steps: A, preparation of polycaprolactone microspheres: (a) preparation of a water phase: adding a surfactant into purified water; the viscosity of the aqueous phase solution is 2-10cP, (b) preparing an organic phase: adding polycaprolactone into an organic solvent, (c) emulsifying and granulating: uniformly mixing the organic phase and the aqueous phase, emulsifying by using a membrane emulsifier, and preparing microspheres, (d) stirring and volatilizing: stirring to completely volatilize the organic solvent, and (e) collecting the microspheres; and B, preparation of a filler composition: (1) weighing a buffer solution, (2) adding a suspension stabilizer and polynucleotide and / or polydeoxynucleotide after filtration, and (3) adding polycaprolactone microspheres into the solution. The filler composition prepared by the invention can stimulate the growth of collagen, and can effectively avoid the defects of aggregation, pinhole blockage, nodule formation and the like during injection.

Description

[0001] This case is a divisional application. The application number of the parent case is 202311046178.5, the application date is 2023.08.18, and the name of the invention is "A filler composition, its preparation method and its application". Technical Field

[0002] The present invention belongs to the field of medical cosmetic materials, and in particular relates to a filler composition, a preparation method thereof and an application thereof. Background Art

[0003] Hyaluronan (Hyaluronan, Hyaluronic Acid), also known as hyaluronic acid, is a glycosaminoglycan composed of disaccharides (D-glucuronic acid and N-acetylglucosamine) and is one of the main components of the extracellular matrix. Hyaluronic acid is widely present in connective tissue, epithelial tissue and neural tissue. Unlike most glycosaminoglycans, hyaluronic acid does not contain sulfur and is formed in the cell membrane rather than in the GABAergic body. Its relative molecular mass can reach 10 6 Magnitude. Hyaluronic acid is recognized as the best moisturizing ingredient today and is widely used in skin care products and cosmetics. Hyaluronic acid gel developed with hyaluronic acid as the main raw material is injected into the superficial dermis of human skin, which can maintain moisture in the skin, promote the supply of nutrients and the excretion of metabolites; promote the proliferation and differentiation of epidermal cells, and remove oxygen free radicals, thereby preventing skin aging and playing a role in beauty and skin care. It is known that the degradation cycle of sodium hyaluronate is short, and the cross-linked sodium hyaluronate is generally 6-12 months. Its short degradation cycle in the body cannot achieve long-term effects.

[0004] PDRN (polydeoxyribonucleotide) helps standardize cell genes, promotes the increase in the number of fibroblasts, and activates secretion, allowing wounds to heal quickly. By utilizing this characteristic, wounds can be quickly restored and wound healing time can be shortened. It can be applied to eliminate scars or fine wrinkles, prevent skin aging, enhance skin elasticity, and other fields. It is a treatment method that essentially promotes skin regeneration.

[0005] As a raw material for skin fillers, polycaprolactone is mostly used to fill wrinkles and depressions in the skin. Polyester materials have the advantages of good biocompatibility, anticoagulability, non-toxicity and low immunity. They can be hydrolyzed and degraded into low molecular weight fragments in a physiological environment. Low molecular weight fragments can be endocytosed by macrophages and degraded within the cells. During the degradation process, they stimulate the host's immune response, attract a large number of macrophages and other immune cells under the skin, lead to the proliferation of fibroblasts and the differentiation of myofibroblasts, synthesize a large amount of collagen, thereby restoring subcutaneous capacity and repairing wrinkles. Polycaprolactone can be 100% absorbed by the human body. Compared with polylactic acid, after implantation into the skin, polycaprolactone is absorbed more slowly than polylactic acid, can promote the production of collagen, and the effect can last for 1 to 4 years.

[0006] Polyester materials are hydrophobic and tend to aggregate to form large particles in dermal fillers. They only show effects 6 to 8 weeks after injection into the skin, and their treatment satisfaction is lower than that of hyaluronic acid fillers, which show effects immediately after treatment.

[0007] Therefore, it is necessary to develop a new polycaprolactone filling composition to improve the current technical problems of polycaprolactone injections, such as slow onset of effect, strong injection irritation, and instability. Summary of the invention

[0008] In view of the deficiencies of the prior art, the present invention provides a filler composition that promotes cell and tissue regeneration, can not only quickly stimulate the regeneration of collagen, but also exhibits high tissue repair properties, prolongs the maintenance effect, prevents skin aging, and enhances skin elasticity.

[0009] One of the objects of the present invention is to provide a filler composition comprising the following components: 20-50 wt % polycaprolactone microspheres, 0.1-3 wt % polynucleotides and / or polydeoxynucleotides, 1-60 wt % suspension stabilizers, and 10-70 wt % buffer.

[0010] In a preferred technical solution of the present invention, the content of polycaprolactone microspheres in the filler composition is 25-40wt%, preferably 25-35wt%.

[0011] In a preferred technical solution of the present invention, the content of polynucleotide and / or polydeoxynucleotide in the filler composition is 0.3-2wt%, preferably 0.5-1wt%.

[0012] In a preferred technical solution of the present invention, the content of the suspension stabilizer in the filler composition is 2-55wt%, preferably 3-50wt%.

[0013] In a preferred technical solution of the present invention, the filler composition optionally contains glycerol, and the content of glycerol is 0.1-1wt%, preferably 0.2-0.8wt%, and more preferably 0.4-0.7wt%.

[0014] In a preferred technical solution of the present invention, the filler composition optionally contains lidocaine, and the content of lidocaine is 0.1-0.5wt%, preferably 0.2-0.4wt%.

[0015] In a preferred technical solution of the present invention, the pH of the composition is 6-8.

[0016] In the preferred technical solution of the present invention, the suspension stabilizer is selected from any one of polyvinyl alcohol, sodium hyaluronate, cross-linked sodium hyaluronate, sodium carboxymethyl cellulose, methyl cellulose, and hydroxypropyl methyl cellulose, or a combination thereof.

[0017] In the preferred technical solution of the present invention, the buffer is selected from any one of phosphate buffer, citrate buffer, sodium chloride buffer or a combination thereof. Preferably, the buffer is sodium chloride or phosphate buffer.

[0018] Another object of the present invention is to provide a method for preparing a filler composition, the filler composition comprising 20-50 wt% polycaprolactone microspheres, 0.1-3 wt% polynucleotides and / or polydeoxynucleotides, 1-60 wt% suspension stabilizers, and 10-70 wt% buffer. The preparation method comprises the following steps: (1) weighing the buffer and filtering it; (2) adding the suspension stabilizer and the polynucleotides and / or polydeoxynucleotides after filtering, and stirring to dissolve them; and (3) adding the polycaprolactone microspheres to the solution, and stirring to evenly disperse the polycaprolactone.

[0019] In the preferred technical solution of the present invention, glycerol is optionally added in step (1) and then mixed evenly and filtered.

[0020] In a preferred technical solution of the present invention, the content of polycaprolactone microspheres in the filler composition is 25-40wt%, preferably 25-35wt%.

[0021] In a preferred technical solution of the present invention, the content of polynucleotide and / or polydeoxynucleotide in the filler composition is 0.3-2wt%, preferably 0.5-1wt%.

[0022] In a preferred technical solution of the present invention, the content of the suspension stabilizer in the filler composition is 2-55wt%, preferably 3-50wt%.

[0023] In a preferred technical solution of the present invention, the filler composition optionally contains glycerol, and the content of glycerol is 0.1-1wt%, preferably 0.2-0.8wt%, and more preferably 0.4-0.7wt%.

[0024] In a preferred technical solution of the present invention, the filler composition optionally contains lidocaine, and the content of lidocaine is 0.1-0.5wt%, preferably 0.2-0.4wt%.

[0025] In the preferred technical solution of the present invention, the suspension stabilizer is selected from any one of polyvinyl alcohol, sodium hyaluronate, cross-linked sodium hyaluronate, sodium carboxymethyl cellulose, methyl cellulose, and hydroxypropyl methyl cellulose, or a combination thereof.

[0026] In the preferred technical solution of the present invention, the buffer is selected from any one of phosphate buffer, citrate buffer, sodium chloride buffer or a combination thereof. Preferably, the buffer is sodium chloride or phosphate buffer.

[0027] Another object of the present invention is to provide an injectable filler, wherein the filler is polycaprolactone microspheres, the particle size of the microspheres is 20-100 μm, and the polycaprolactone microspheres have a rough surface or a non-smooth surface.

[0028] In a preferred technical solution of the present invention, the rough surface or non-smooth surface of the polycaprolactone microspheres has irregular pore sizes.

[0029] In the preferred technical solution of the present invention, the particle size of the microspheres is preferably 20-70 μm, more preferably 25-50 μm.

[0030] In the preferred technical solution of the present invention, the molecular weight of the polycaprolactone is 5000-30000.

[0031] In the preferred technical solution of the present invention, the molecular weight of the polycaprolactone is 10000-25000.

[0032] In the preferred technical solution of the present invention, the molecular weight of the polycaprolactone is 15000-22000.

[0033] Another object of the present invention is to provide a method for preparing an injection filler, comprising the following steps:

[0034] (1) Preparation of aqueous phase: Add surfactant into purified water, stir and dissolve;

[0035] (2) Preparation of organic phase: Add the injectable filler into the organic solvent and stir until completely dissolved;

[0036] (3) Emulsification granulation: The organic phase and the aqueous phase are mixed evenly, stirred, and emulsified using a membrane emulsifier to prepare microspheres;

[0037] (4) Stirring and volatilizing: Stirring to evaporate all the organic solvent;

[0038] (5) Microsphere collection: Collect the suspension, filter, wash, and dry to obtain injectable filler microspheres.

[0039] In the preferred technical solution of the present invention, the surfactant is any one of polyvinyl alcohol, methyl cellulose, polyoxyethylene, polyoxypropylene, polyethyleneimine, polystyrene, polyoxysilane, hydroxypropyl cellulose or a combination thereof.

[0040] In the preferred technical solution of the present invention, the organic solvent is any one of acetone, ethanol, ethyl acetate, chloroform, and dichloromethane, or a combination thereof.

[0041] In the preferred technical solution of the present invention, the ratio of the surfactant to purified water is 10-50 g / L, preferably 15-40 g / L, and more preferably 20-30 g / L.

[0042] In a preferred technical solution of the present invention, the viscosity of the aqueous solution is 2-10 cP, preferably 4-8 cP.

[0043] In the preferred technical scheme of the present invention, during the preparation of the aqueous phase solution, purified water is added to the reaction kettle, mechanical stirring is turned on, and the rotation power is adjusted to 10-50 Hz, preferably 20-30 Hz; the surfactant is slowly added to the purified water, heated to 60-120° C., stirred for 2 to 6 hours, and cooled to room temperature after the surfactant is completely dissolved.

[0044] In a preferred technical solution of the present invention, the ratio of the injectable filler to the organic solvent is 100-300 g / L, preferably 150-250 g / L.

[0045] In a preferred technical solution of the present invention, during the preparation of the organic phase, the injectable filler is added to the organic solvent and stirred to completely dissolve it.

[0046] In the preferred technical scheme of the present invention, in the emulsification step, the mechanical stirring power is adjusted to 10-50 Hz, the organic phase is slowly added to the aqueous phase, and after stirring for 3-10 minutes, the pressure is adjusted to 0.5-1 bar using a membrane emulsifier, and a 30 μm membrane tube is used for rapid membrane emulsification.

[0047] In the preferred technical scheme of the present invention, in the stirring and volatilization step, the emulsified sample is transferred to a reaction kettle, the mechanical stirring power is maintained at 10 to 50 Hz, nitrogen / compressed air is used to replace the gas in the kettle, a flow rate of 1 to 20 m / s is maintained, and stirring is performed for 12 to 48 hours to completely volatilize the organic solvent.

[0048] In a preferred technical solution of the present invention, in the microsphere collection step, the microspheres are collected by centrifugation, sedimentation, filtration or a combination thereof.

[0049] In the preferred technical solution of the present invention, in the microsphere collection step, after the organic solvent evaporates, the suspension in the reaction kettle is collected, and the solid microspheres are separated using a centrifuge. The centrifuge speed is 2000-5000 rpm, and the centrifugation time is 2-8 min. Preferably, the centrifuge speed is 3000-4000 rpm, and the centrifugation time is 3-6 min.

[0050] In the preferred technical solution of the present invention, in the microsphere collection step, the collected microspheres are taken, washed with purified water and ethanol, collected after centrifugation, and dried to obtain the microspheres.

[0051] In the preferred technical solution of the present invention, the drying is carried out at 30-60°C for 10-30 hours, preferably at 30-40°C for 15-24 hours.

[0052] In the preferred technical solution of the present invention, the drying is air-blast drying or freeze-drying.

[0053] In the preferred technical solution of the present invention, in the cleaning step, purified water is used for cleaning 2-5 times, and ethanol is used for cleaning 1-2 times.

[0054] Another object of the present invention is to provide a filler composition for use as an injectable filler, wherein the filler composition comprises the following components: 20-50wt% polycaprolactone microspheres, 0.1-3wt% polynucleotides and / or polydeoxynucleotides, 1-60wt% suspension stabilizer, and 10-70wt% buffer.

[0055] In a preferred technical solution of the present invention, the filling site of the injection filler is selected from any one of the face, neck, abdomen, chest, buttocks, thighs, calves, upper arms, and lower arms, or a combination thereof, and the preferred injection filling site is the face.

[0056] In a preferred technical solution of the present invention, the injectable filler is used to improve any one or a combination of facial emaciation, fat atrophy, sunken cheeks, sunken eye sockets, and skin wrinkles in patients.

[0057] Another object of the present invention is to provide a combined use of a filler composition, wherein the filler composition is used in combination with any one of other types of injectable fillers, anti-inflammatory agents, anti-allergic agents or a combination thereof; the filler composition comprises the following components: 20-50wt% polycaprolactone microspheres, 0.1-3wt% polynucleotides and / or polydeoxynucleotides, 1-60wt% suspension stabilizers, and 10-70wt% buffer.

[0058] In the preferred technical solution of the present invention, the other types of injectable fillers are selected from any one of polymethyl methacrylate, polyacrylamide, silicone, autologous fat, or a combination thereof.

[0059] In the preferred technical solution of the present invention, the anti-inflammatory agent is selected from any one of steroidal anti-inflammatory agents and non-steroidal anti-inflammatory agents or a combination thereof.

[0060] In the preferred technical solution of the present invention, the steroid anti-inflammatory agent is selected from any one of fluocinolone, hydrocortisone, betamethasone or a combination thereof.

[0061] In the preferred technical scheme of the present invention, the non-steroidal anti-inflammatory agent is selected from any one of aspirin, magnesium salicylate, sodium salicylate, choline magnesium salicylate, diflunisal, disalsalate, ibuprofen, indomethacin, flurbiprofen, phenoxyibuprofen, naproxen, nabumetone, piroxicam, phenylbutazone, diclofenac, fenprofen, ketoprofen, ketorolac, tetraclofenamic acid, sulindac, and tolmetin, or a combination thereof.

[0062] In the preferred technical scheme of the present invention, the antiallergic agent is selected from any one of diphenhydramine, promethazine, chlorpheniramine, sodium cromoglycate, ketotifen, betahistine, montelukast, zafirlukast, salbutamol, calcium gluconate, and adrenal glucocorticoids, or a combination thereof.

[0063] The present invention adopts gel permeation chromatography (GPC) method (Chinese Pharmacopoeia 2020 Edition Part Four General Rules 0514) to detect the molecular weight of the copolymer, as shown below:

[0064] Take an appropriate amount of the test sample solution, weigh it accurately, add chloroform to dissolve and dilute it to make a solution containing about 10 mg per 1 ml, filter it with a 0.22 μm organic filter membrane, and take the filtrate.

[0065] For the reference solution, take appropriate amounts of 5-6 polystyrene reference substances of different molecular weights, weigh them accurately, dissolve them in chloroform and dilute them to make a solution containing about 2 mg of each reference substance per 1 ml, dissolve them and mix them evenly.

[0066] The chromatographic conditions are as follows: a gel column (Styragel HR 4E THF, 7.8×300 mm or a column of equivalent performance); the mobile phase is chloroform; the flow rate is 0.7 ml per minute; the detector is a differential refractive index detector; the column temperature is 35° C.; and the injection volume is 10 μL.

[0067] Determination method: Precisely measure the test solution and reference solution, inject them into the liquid chromatograph, and record the chromatogram. The regression equation of the reference solution and the weight average molecular weight (Mw), number average molecular weight (Mn) and molecular weight distribution (Mw / Mn) of the test sample are calculated by GPC software.

[0068] The present invention adopts an electronic tensile testing machine to measure the pushing force: the use speed is 30 mm / min, the stopping force value is 250.00 N, and the stopping displacement is 15.0 mm.

[0069] The present invention adopts Kinexus Pro + Rheometer Shear Viscosity and Rheological Properties.

[0070] Shear viscosity test: Select rotor model: PU20 L1917 SS, set gap: 0.3mm; after loading the sample, select shear rate scanning program, set temperature to 25℃, and start shear rate to 0.01S -1 , the end shear rate is 100.00 -1 The scanning time is 3 min, and the number of sampling points for each order of magnitude is 20. The shear rate is 1S -1 Shear viscosity at .

[0071] Select the rotor model: PU20 L1917 SS, set the gap: 0.3mm. After loading the sample, select the frequency table (strain control), set the temperature to 25℃, the starting frequency to 10HZ, the ending frequency to 0.1HZ, the shear strain to 0.5%, and the number of sampling points for each order of magnitude is 10, and obtain G' and G" at a frequency of 1HZ.

[0072] Unless otherwise specified, when the present invention relates to the percentage between liquids, the percentage is volume / volume percentage; when the present invention relates to the percentage between liquids and solids, the percentage is volume / weight percentage; when the present invention relates to the percentage between solids and liquids, the percentage is weight / volume percentage; the rest are weight / weight percentages.

[0073] Compared with the prior art, the present invention has the following beneficial technical effects:

[0074] 1. The present invention adds polynucleotides and / or polydeoxynucleotides to the polycaprolactone injection to promote cell and tissue regeneration, show high tissue repair properties, prolong the maintenance effect, prevent skin aging, and enhance skin elasticity. The present invention scientifically screens the content of each component so that the polycaprolactone microspheres can be evenly and stably suspended in the injection solution, and the polycaprolactone microspheres have a uniform particle size distribution, avoiding defects such as aggregation, pinhole clogging, and nodule formation during injection.

[0075] 2. The polycaprolactone microspheres in the present invention have a rough surface or a non-smooth surface. Compared with a smooth surface, the polycaprolactone microspheres with a rough surface and a porous structure have a higher specific surface area, thereby increasing the contact area with cells, improving the adhesion ability and retention time of cells, and improving the affinity of the polycaprolactone microspheres, which can stimulate and accelerate the growth of collagen.

[0076] 3. The present invention scientifically screens the dosage of each component in the process of preparing polycaprolactone microspheres, and obtains regular, rough-surfaced, uniform-size spherical polycaprolactone microparticles by combining a rapid membrane emulsification method on the basis of controlling the viscosity of the aqueous solution. The polycaprolactone microspheres obtained by the preparation method of the present invention have a narrow particle size distribution range and a uniform particle size, thereby improving the yield of injectable polycaprolactone microspheres. BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Figure 1 (a) is a sedimentation trend diagram of Example 5;

[0078] Figure 1(b) is a sedimentation trend diagram of Comparative Example 1. DETAILED DESCRIPTION

[0079] The present invention will be specifically described below in conjunction with embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention and are not intended to limit the essence of the present invention.

[0080] Example 1: Preparation of polycaprolactone microspheres

[0081] ① Preparation of aqueous phase: Add 5L of purified water to a 10L reactor, start mechanical stirring, and adjust the rotation power to 25Hz; slowly add 100g of polyvinyl alcohol, heat to 80℃, stir for 5 hours, and cool to room temperature (viscosity 5cP) after the polyvinyl alcohol is completely dissolved;

[0082] ② Preparation of organic phase: Add 500 mL of dichloromethane to a beaker, weigh 100 g of polycaprolactone with a Mn of 20,000, add it to the beaker, and stir to completely dissolve it;

[0083] ③ Emulsification: adjust the mechanical stirring power to 20 Hz, slowly add the above organic phase into the aqueous phase, stir for 5 min, adjust the pressure to 0.8 bar using a membrane emulsifier, and use a 30 μm membrane tube for rapid membrane emulsification;

[0084] ④ Stirring and volatilizing: transfer the emulsified sample to a 10L reactor, maintain the mechanical stirring power at 20Hz, use nitrogen / compressed air to replace the gas in the reactor, maintain a flow rate of 20m / s, and stir for 24 hours to completely volatilize the organic solvent;

[0085] ⑤ Microsphere collection: After the dichloromethane evaporates, collect the suspension in the reactor, use a centrifuge to separate the solid microspheres (3500rpm, 5min), and wash with purified water and ethanol (wash with purified water 3 times, wash with ethanol once) and centrifuge. Put the sample obtained after centrifugation into a forced air drying oven, adjust the temperature to 40°C, and dry for 20 hours; obtain microspheres with a particle size of 20-80μm.

[0086] Example 2 Preparation of polycaprolactone microspheres

[0087] ① Preparation of aqueous phase: Add 5L of purified water to a 10L reactor, start mechanical stirring, and adjust the rotation power to 25Hz; slowly add 120g of polyvinyl alcohol, heat to 80℃, stir for 6 hours, and cool to room temperature (viscosity 5.8cP) after the polyvinyl alcohol is completely dissolved;

[0088] ② Preparation of organic phase: Add 500 mL of chloroform to a beaker, weigh 110 g of polycaprolactone with a Mn of 22,000 and add it to the beaker, and stir to completely dissolve it;

[0089] ③ Emulsification: Adjust the mechanical stirring power to 25 Hz, slowly add the above organic phase into the aqueous phase, stir for 5 min, adjust the pressure to 1 bar using a membrane emulsifier, and use a 30 μm membrane tube for rapid membrane emulsification;

[0090] ④ Stirring and volatilizing: transfer the emulsified sample to a 10L reactor, maintain the mechanical stirring power at 15-25Hz, use nitrogen / compressed air to replace the gas in the reactor, maintain a flow rate of 15m / s, and stir for 30 hours to completely volatilize the organic solvent;

[0091] ⑤ Microsphere collection: After the chloroform evaporates, collect the suspension in the reactor, use a centrifuge to separate the solid microspheres (3500rpm, 5min), and wash with purified water and ethanol (wash with purified water 3 times, wash with ethanol once) and centrifuge. Put the sample obtained after centrifugation into a forced air drying oven, adjust the temperature to 40°C, and dry for 20 hours; obtain microspheres with a particle size of 20-80μm.

[0092] Example 3 Preparation of polycaprolactone microspheres

[0093] ① Preparation of aqueous phase: Add 5L of purified water to a 10L reactor, start mechanical stirring, and adjust the rotation power to 25Hz; slowly add 80g of polyvinyl alcohol, heat to 80℃, stir for 4 hours, and cool to room temperature (viscosity 4.7cP) after the polyvinyl alcohol is completely dissolved;

[0094] ② Preparation of organic phase: Add 500 mL of dichloromethane to a beaker, weigh 80 g of polycaprolactone with a Mn of 20,000 and add it to the beaker, and stir to completely dissolve it;

[0095] ③ Emulsification: adjust the mechanical stirring power to 20 Hz, slowly add the above organic phase into the aqueous phase, stir for 5 min, adjust the pressure to 0.8 bar using a membrane emulsifier, and use a 30 μm membrane tube for rapid membrane emulsification;

[0096] ④ Stirring and volatilizing: transfer the emulsified sample to a 10L reactor, maintain the mechanical stirring power at 20Hz, use nitrogen / compressed air to replace the gas in the reactor, maintain a flow rate of 20m / s, and stir for 30 hours to completely volatilize the organic solvent;

[0097] ⑤ Microsphere collection: After the dichloromethane evaporates, collect the suspension in the reactor, filter the microspheres, wash them with purified water and ethanol (wash them 3 times with purified water and once with ethanol), and centrifuge them. Put the sample obtained after centrifugation into a forced air drying oven, adjust the temperature to 40°C, and dry it for 20 hours; obtain microspheres with a particle size of 20-80 μm.

[0098] Example 4 Preparation of polycaprolactone microspheres

[0099] ① Preparation of aqueous phase: Add 5L of purified water to a 10L reactor, start mechanical stirring, and adjust the rotation power to 25Hz; slowly add 60g of methylcellulose, heat to 80℃, stir for 4 hours, and cool to room temperature (viscosity 7cP) after the methylcellulose is completely dissolved;

[0100] ② Preparation of organic phase: Add 500 mL of dichloromethane to a beaker, weigh 100 g of polycaprolactone with a Mn of 18,000 and add it to the beaker, and stir to completely dissolve it;

[0101] ③ Emulsification: adjust the mechanical stirring power to 20 Hz, slowly add the above organic phase into the aqueous phase, stir for 5 min, adjust the pressure to 0.8 bar using a membrane emulsifier, and use a 30 μm membrane tube for rapid membrane emulsification;

[0102] ④ Stirring and volatilizing: transfer the emulsified sample to a 10L reactor, maintain the mechanical stirring power at 20Hz, use nitrogen / compressed air to replace the gas in the reactor, maintain a flow rate of 20m / s, and stir for 28 hours to completely volatilize the organic solvent;

[0103] ⑤ Microsphere collection: After the dichloromethane evaporates, collect the suspension in the reactor, use a centrifuge to separate the solid microspheres (3500rpm, 5min), and wash with purified water and ethanol (wash with purified water 3 times, wash with ethanol once) and centrifuge. Put the sample obtained after centrifugation into a forced air drying oven, adjust the temperature to 40°C, and dry for 20 hours; obtain microspheres with a particle size of 20-80μm.

[0104] Example 5 Preparation of polycaprolactone composition preparation

[0105] Weigh 627g of phosphate buffer, add 7g of glycerol, mix well and filter with 0.22μm; add 30g of sodium carboxymethyl cellulose and 6g of polydeoxynucleotide to the filtrate in turn, stir mechanically for 3 hours to completely dissolve them, after sodium carboxymethyl cellulose and polydeoxynucleotide are completely dissolved, add 330g of polycaprolactone microspheres of Example 1 to the solution, stir to evenly disperse the polycaprolactone; after the sample is evenly dispersed, the sample is obtained. The method of the present invention is used to detect the pushing force, viscoelasticity and shear viscosity.

[0106] Average pushing force: 25.630N (27G*12MM needle speed 30mm / min)

[0107] Shear viscosity: 280.4pa·s

[0108] Viscoelasticity: G': 946.7pa G': 713.5pa at 1HZ

[0109] pH: 7.40

[0110] Example 6 Preparation of polycaprolactone composition preparation

[0111] Weigh 164g of phosphate buffer and filter it through 0.22μm; add 450g of cross-linked sodium hyaluronate gel (20mg / g), 50g of non-cross-linked sodium hyaluronate solution (20mg / g) and 6g of polydeoxynucleotide to the filtered filtrate in turn, stir mechanically for 3 hours to completely dissolve them, after the sodium hyaluronate and polydeoxynucleotide are completely dissolved, add 330g of polycaprolactone microspheres of Example 2 to the solution, stir to evenly disperse the polycaprolactone; after the sample is evenly dispersed, the sample is obtained. The method of the present invention is used to detect the pushing force, viscoelasticity, and shear viscosity.

[0112] Average pushing force: 15.684N (27G*12MM needle speed 30mm / min)

[0113] Shear viscosity: 184.2pa·s

[0114] Viscoelasticity: G': 188.25pa G': 43.65pa at 1HZ

[0115] pH: 7.14

[0116] Example 7 Preparation of polycaprolactone composition preparation

[0117] Weigh 158g of phosphate buffer, add 7g of glycerol, mix well and filter with 0.22μm; add 500g of cross-linked sodium hyaluronate gel (20mg / g) and 5g of polydeoxynucleotide to the filtrate in turn, stir mechanically for 3 hours to completely dissolve it, after the cross-linked sodium hyaluronate and polydeoxynucleotide are completely dissolved, add 330g of polycaprolactone microspheres of Example 3 to the solution, stir to evenly disperse the polycaprolactone; after the sample is evenly dispersed, dispense it into a 1ml prefilled syringe. The method of the present invention is used to detect the pushing force, viscoelasticity, and shear viscosity.

[0118] Average pushing force: 17.340N (27G*12MM needle speed 30mm / min)

[0119] Shear viscosity: 112.6pa·s

[0120] Viscoelasticity: G': 232.1pa G': 33.32pa at 1HZ

[0121] pH: 7.18

[0122] Example 8 Preparation of polycaprolactone composition preparation

[0123] Weigh 625g of phosphate buffer, add 7g of glycerol, mix well and filter with 0.22μm; add 30g of sodium carboxymethylcellulose, 5g of polydeoxynucleotide and 3g of lidocaine to the filtrate in turn after filtration, stir mechanically for 3 hours to completely dissolve them, after sodium carboxymethylcellulose and polydeoxynucleotide are completely dissolved, add 330g of polycaprolactone microspheres of Example 4 to the solution, stir to evenly disperse the polycaprolactone; after the sample is evenly dispersed, dispense into 1ml prefilled syringes. The method of the present invention is used to detect pushing force, viscoelasticity and shear viscosity.

[0124] Average pushing force: 24.652N (27G*12MM needle speed 30mm / min)

[0125] Shear viscosity: 304.6pa·s

[0126] Viscoelasticity: G': 1056.2pa G": 845.8pa at 1HZ

[0127] pH: 7.35

[0128] Comparative Example 1 Preparation of polycaprolactone composition preparation

[0129] Polycaprolactone microspheres were prepared using the method of Example 1 of CN106075436B:

[0130] 20 g of PCL particles with an Mn of 10,000 were dissolved in DCM to form a DCM solution containing 10-20 wt% PCL, and the solution was dispersed in 1000 ml of water containing 5% MC, stirred, filtered, washed, and dried to obtain polycaprolactone microspheres with a particle size of 10-180 μm.

[0131] Weigh 633 g of phosphate buffer, add 7 g of glycerol, mix well and filter with 0.22 μm; add 30 g of sodium carboxymethyl cellulose to the filtrate after filtration, stir mechanically for 3 hours to completely dissolve it, after the sodium carboxymethyl cellulose is completely dissolved, add 330 g of polycaprolactone microspheres to the solution, stir to evenly disperse the polycaprolactone; after the sample is evenly dispersed, dispense into 1 ml prefilled syringes.

[0132] Test Example 1

[0133] The polycaprolactone composition preparation prepared in Example 5 and the polycaprolactone composition preparation prepared in Comparative Example 1 were placed to observe the sedimentation and aggregation of the polycaprolactone particles.

[0134] The polycaprolactone composition preparation was evenly divided into three layers: upper, middle and lower. Samples were taken from each layer at 0 day, 1 month, 2 months and 3 months respectively to detect the number of polycaprolactone particles (repeated three parallel samples). The results are shown in Figure 1.

[0135] The results in Figure 1 show that the particles in each layer of Comparative Example 1 are unevenly distributed. As the storage time increases, the particles in the lower layer gradually increase, while the particles in the upper and middle layers gradually decrease. The number of particles in each layer of the polycaprolactone composition prepared in Example 5 did not change significantly within 3 months, and had good dispersion stability.

[0136] The above description is only an example of the implementation mode of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the technical principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing a filler composition, characterized in that: The steps include: A. Preparation of polycaprolactone microspheres (a) Preparation of aqueous phase: Add surfactant to purified water, stir, and dissolve; the viscosity of the aqueous phase solution is 2-10 cP; (b) Preparation of organic phase: Add polycaprolactone to an organic solvent and stir until completely dissolved; (c) Emulsification granulation: The organic phase and the aqueous phase are mixed uniformly, stirred, and emulsified using a membrane emulsifier to prepare microspheres; (d) stirring and volatilizing: stirring to evaporate all the organic solvent; (e) microsphere collection: collecting the suspension, filtering, washing, and drying to obtain polycaprolactone microspheres; B. Preparation of Filler Compositions (1) Weigh the buffer solution; (2) adding a suspension stabilizer and polynucleotides and / or polydeoxynucleotides after filtration and stirring to dissolve them; (3) adding polycaprolactone microspheres into the solution and stirring to uniformly disperse the polycaprolactone microspheres to obtain; The proportions of the components in the composition are as follows: 20-50 wt% polycaprolactone microspheres, 0.1-3 wt% polynucleotide and / or polydeoxynucleotide, 1-60 wt% suspension stabilizer, and 10-70 wt% buffer.

2. The method for preparing the filler composition according to claim 1, wherein: The filler composition satisfies at least one of the following conditions (1) to (10): (1) The content of polycaprolactone microspheres in the filler composition is 25-40 wt%; (2) the content of polynucleotides and / or polydeoxynucleotides in the filler composition is 0.3-2 wt %; (3) The content of the suspension stabilizer in the filler composition is 2-55 wt%; (4) The filler composition contains glycerol, and the content of glycerol is 0.1-1wt%; (5) The filler composition contains lidocaine, and the content of lidocaine is 0.1-0.5 wt%; (6) The suspension stabilizer is selected from any one of polyvinyl alcohol, sodium hyaluronate, cross-linked sodium hyaluronate, sodium carboxymethyl cellulose, methyl cellulose, and hydroxypropyl methyl cellulose, or a combination thereof; (7) The buffer is selected from any one of phosphate buffer and citrate buffer or a combination thereof; (8) The pH of the filler composition is 6-8; (9) The particle size of the polycaprolactone microspheres is 20-70 μm; (10) The molecular weight of the polycaprolactone is 5000-30000.

3. The method for preparing the filler composition according to claim 2, wherein: The filler composition satisfies at least one of the following conditions (1) to (7): (1) The content of polycaprolactone microspheres in the filler composition is 25-35 wt%; (2) the content of polynucleotides and / or polydeoxynucleotides in the filler composition is 0.5-1 wt%; (3) The content of the suspension stabilizer in the filler composition is 3-50 wt%; (4) The filler composition contains glycerol, and the content of glycerol is 0.2-0.8wt%, preferably 0.4-0.7wt%; (5) The filler composition contains lidocaine, and the content of lidocaine is 0.2-0.4 wt%; (6) The particle size of the polycaprolactone microspheres is 25-50 μm; (7) The molecular weight of the polycaprolactone is 10,000-25,000.

4. The method for preparing a filler composition according to any one of claims 1 to 3, characterized in that: The preparation method of the polycaprolactone microspheres satisfies at least one of the following conditions (1) to (16): (1) The molecular weight of the polycaprolactone is 15000-22000; (2) The surfactant is any one of polyvinyl alcohol, methyl cellulose, polyoxyethylene, polyoxypropylene, polyethyleneimine, polystyrene, polyoxysilane, and hydroxypropyl cellulose, or a combination thereof; (3) The organic solvent is any one of acetone, ethanol, ethyl acetate, chloroform, and dichloromethane, or a combination thereof; (4) The ratio of the surfactant to purified water is 10-50 g / L; (5) The viscosity of the aqueous solution is 4-8 cP; (6) During the preparation of the aqueous phase solution, purified water is added to the reactor, mechanical stirring is started, and the rotation power is adjusted to 10-50 Hz; the surfactant is slowly added to the purified water, heated to 60-120° C., stirred for 2-6 hours, and cooled to room temperature after the surfactant is completely dissolved; (7) The ratio of the injectable filler to the organic solvent is 100-300 g / L; (8) During the preparation of the organic phase, the injectable filler is added to the organic solvent and stirred to completely dissolve it; (9) In the emulsification step, the mechanical stirring power is adjusted to 10-50 Hz, the organic phase is slowly added to the aqueous phase, and after stirring for 3-10 minutes, the pressure is adjusted to 0.5-1 bar using a membrane emulsifier, and a 30 μm membrane tube is used for rapid membrane emulsification; (10) In the stirring and volatilization step, the emulsified sample is transferred to a reactor, the mechanical stirring power is maintained at 10-50 Hz, nitrogen / compressed air is used to replace the gas in the reactor, and the flow rate is maintained at 1-20 m / s. The sample is stirred for 12-48 hours to completely volatilize the organic solvent; (11) In the microsphere collection step, the microspheres are collected by centrifugation, sedimentation, filtration, or a combination thereof; (12) In the microsphere collection step, after the organic solvent evaporates, the suspension in the reactor is collected and the solid microspheres are separated using a centrifuge at a speed of 2000-5000 rpm and a centrifugation time of 2-8 min. (13) In the microsphere collection step, the collected microspheres are washed with purified water and ethanol, collected after centrifugation, and dried to obtain; (14) The drying step is performed at 30 to 60° C. for 10 to 30 hours; (15) The drying is air drying or freeze drying; In the cleaning step described in (16), the purified water is used for cleaning 2-5 times, and the ethanol is used for cleaning 1-2 times.

5. The method for preparing the filler composition according to claim 4, wherein: The preparation method of the polycaprolactone microspheres satisfies at least one of the following conditions (1) to (5): (1) The ratio of the surfactant to purified water is 15-40 g / L, preferably 20-30 g / L; (2) During the preparation of the aqueous phase solution, purified water was added to the reactor, mechanical stirring was started, and the rotation power was adjusted to 20-30 Hz; (3) The ratio of the injectable filler to the organic solvent is 150-250 g / L; (4) In the microsphere collection step, after the organic solvent evaporates, the suspension in the reaction vessel is collected and the solid microspheres are separated using a centrifuge at a speed of 3000-4000 rpm and a centrifugation time of 3-6 min; (5) The drying step is performed at 30-40°C for 15-24 hours.

6. The method for preparing a filler composition according to claim 1, characterized in that: In the step (1), glycerol is added, mixed evenly, and filtered.

7. A filler composition, characterized in that The filler composition is prepared by the method for preparing the filler composition according to any one of claims 1 to 6.

8. Use of the filler composition prepared by the preparation method according to any one of claims 1 to 6 as an injectable filler; Preferably, the injection site of the filler is selected from any one of the face, neck, abdomen, chest, buttocks, thighs, calves, upper arms, lower arms, or a combination thereof, and preferably the face. Preferably, the injectable filler is used to improve any one of or a combination of facial emaciation, fat atrophy, sunken cheeks, sunken eye sockets, and skin wrinkles in a patient.

9. A combined use method of the filler composition prepared by the preparation method according to any one of claims 1 to 6, characterized in that: The filler composition is used in combination with any one of other types of injectable fillers, anti-inflammatory agents, anti-allergic agents, or a combination thereof.

10. The combined method according to claim 9, wherein The combined method satisfies at least one of the following conditions (1) to (3): (1) The other types of injectable fillers are selected from any one of polymethyl methacrylate, polyacrylamide, silicone, and autologous fat, or a combination thereof; (2) The anti-inflammatory agent is selected from any one of steroidal anti-inflammatory agents and non-steroidal anti-inflammatory agents or a combination thereof; preferably, the steroidal anti-inflammatory agent is selected from any one of fluocinolone, hydrocortisone, and betamethasone or a combination thereof; preferably, the non-steroidal anti-inflammatory agent is selected from any one of aspirin, magnesium salicylate, sodium salicylate, choline magnesium salicylate, diflunisal, disalsalate, ibuprofen, indomethacin, flurbiprofen, phenoxyibuprofen, naproxen, nabumetone, piroxicam, phenylbutazone, diclofenac, fenprofen, ketoprofen, ketorolac, tetraclofenamic acid, sulindac, and tolmetin or a combination thereof; (3) The antiallergic agent is selected from any one of diphenhydramine, promethazine, chlorpheniramine, sodium cromoglycate, ketotifen, betahistine, montelukast, zafirlukast, salbutamol, calcium gluconate, and adrenal glucocorticoids, or a combination thereof.

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