Method for producing active microparticles according to characteristics of drug, active microparticles produced thereby, and method for producing long-lasting formulation comprising drug using active microparticles

Abstract

The present invention relates to a method for producing active microparticles according to characteristics of a drug, active microparticles produced thereby, and a method for producing a long-lasting formulation comprising a drug using the active microparticles. The active microparticles can bind a drug required to be provided in a long-lasting formulation, can be used as a long-lasting formulation by binding the drug therein, and can be produced as customized active microparticles according to various characteristics of the drug. In addition, a drug required to be provided as a long-lasting formulation can be produced as a long-lasting formulation by using the active microparticles of the present invention without the need for separate equipment for producing the drug as a long-lasting formulation.

Description

Method for producing active microparticles according to the characteristics of a drug, active microparticles produced by the method for producing the same, and method for producing a long-acting formulation comprising a drug using the active microparticles

[0001] The present invention relates to a method for producing active microparticles according to the characteristics of a drug, active microparticles produced by the method for producing the same, and a method for producing a long-acting formulation comprising a drug using the active microparticles.

[0002] The main methods for manufacturing drug delivery microspheres using polymers include solvent evaporation (reference: N. Wakiyama et al., Chem. Pharm. Bull., 30(7), 2621-2628, 1982), solvent extraction (reference: JM Ruiz et al., Int. J. Pharm., 49, 69-77, 1989), phase separation (reference: N. Nihant et al., J. Controlled Release, 35, 117-125, 1995), coacervation (reference: JC Leroux et al., Int. Symp. Control. Rel. Bioact. Mater., Controlled Release Society, Inc., 21 #1118, 1994), salting out (reference: B. Gander et al., J. Microencapsulation, 12(1), 83-97, 1995) and spray drying method (Reference: R. Arshady et al., Polym. Eng. Sci., 30(15), 915-924, 1990). The final properties of microspheres, such as particle size, drug loading ratio, and drug release characteristics, are greatly affected by the manufacturing method. Therefore, an appropriate manufacturing method should be selected by considering not only the properties of the polymer and the drug, but also the properties that the manufactured microspheres must have.

[0003] Using the manufacturing method described above, various drugs requiring long-acting formulations were developed in the form of microparticles using biodegradable polymers. However, in the case of microparticles containing the biodegradable polymer and the drug, separate manufacturing equipment was required to manufacture long-acting formulations using the manufacturing method exemplified above.

[0004] In order to apply various methods for manufacturing drug-containing microspheres using not only the solvent evaporation method, solvent extraction method, phase separation method, and spray drying method, but also the membrane method and microfluidic method, separate manufacturing facilities are required. In the case of existing pharmaceutical companies, the introduction of separate manufacturing facilities was required to develop the long-acting formulation.

[0005] As various drugs become available for production and sale due to patent expiration, development of long-acting formulations is progressing more actively. However, as mentioned above, development of such long-acting formulations requires separate manufacturing facilities, which poses a problem that prevents many companies from developing long-acting formulations.

[0006] Accordingly, there is a need to develop a method for manufacturing drugs in long-acting formulations using existing injection equipment without the need for the introduction of separate manufacturing equipment.

[0007] [Prior Art Literature]

[0008] [Patent Document]

[0009] KR 10-2004-0042152 A1

[0010] The purpose of the present invention is to provide a method for producing active microparticles according to the characteristics of a drug, active microparticles produced by the method for producing the same, and a method for producing a long-acting formulation comprising a drug using the active microparticles.

[0011] Another object of the present invention is to provide an active microparticle capable of binding a drug that needs to be provided as a long-acting formulation, a method for manufacturing a customized active microparticle according to the characteristics of various drugs by binding the drug inside the active microparticle to be used as a long-acting formulation, and an active microparticle manufactured by the method.

[0012] Another object of the present invention is to provide a method for producing a long-acting formulation using the active microparticles of the present invention, without requiring separate equipment for producing a long-acting formulation of a drug that needs to be provided as a long-acting formulation using the active microparticles of the present invention.

[0013] In order to achieve the above-described purpose, the present invention relates to a method for producing active microparticles according to the characteristics of a drug, comprising the steps of: dissolving a biodegradable polymer in an organic solvent to produce an oily solution; dissolving a surfactant in water to produce an aqueous solution; producing an emulsion using the oily solution and the aqueous solution; and freeze-drying the emulsion to produce active microparticles.

[0014] Additionally, the oil solution may additionally contain an activator.

[0015] Additionally, the activator can be prepared as an oily solution by dissolving it in an organic solvent together with a biodegradable polymer.

[0016] Additionally, the activator can be prepared as a solution containing the activator by dissolving it in distilled water, and the first emulsion can be formed by mixing it in an oily solution in which a biodegradable polymer is dissolved in an organic solvent.

[0017] Additionally, the activator may be a channel forming agent and / or a binding activator.

[0018] Additionally, the channel forming agent may be selected from the group consisting of polyvinyl alcohol (PVA), ammonium bicarbonate (ABC), bovine serum albumin (BSA), trehalose, ethanol, pluronic, calcium chloride (CaCl2), surfactants, and mixtures thereof.

[0019] Additionally, the binding activator may be selected from the group consisting of polyethyleneimine (PEI), protamine sulfate, bovine serum albumin (BSA), poly-L-lysine, calcium chloride (CaCl2), and mixtures thereof.

[0020] Another invention for achieving the above-described purpose relates to active microparticles manufactured by the above-described manufacturing method.

[0021] Another invention for achieving the above-described object relates to a method for producing a long-acting formulation containing a drug using active microparticles, comprising the steps of: adding the active microparticles to a buffer solution and stirring to produce a suspension solution containing the active microparticles; and adding a drug to the suspension solution containing the active microparticles and stirring to bind the drug to the active microparticles.

[0022] In addition, after the step of binding a drug to the active microparticles, an additional step of incubating at a temperature higher than the Tg temperature of the biodegradable polymer included in the active microparticles for 1 to 5 hours may be performed.

[0023] Additionally, the condition above the Tg temperature may be 40°C or higher.

[0024] Additionally, by the above incubating step, the pores formed on the outside of the active microparticles can be closed by self-healing.

[0025] The present invention is an active microparticle capable of binding a drug that needs to be provided as a long-acting formulation, and can be used as a long-acting formulation by binding the drug inside, and can be manufactured as a customized active microparticle according to the characteristics of various drugs.

[0026] In addition, by using the active microparticles, a drug that needs to be provided as a long-acting formulation can be manufactured as a long-acting formulation using the active microparticles of the present invention without requiring separate equipment to manufacture the long-acting formulation.

[0027] Figure 1 shows the results of particle size analysis of active fine particles according to one embodiment of the present invention.

[0028] Figure 2 is an SEM analysis photograph of active microparticles according to one embodiment of the present invention.

[0029] Figure 3 shows the results of particle size analysis of active fine particles according to one embodiment of the present invention.

[0030] Figure 4 is an SEM analysis photograph of active microparticles according to one embodiment of the present invention.

[0031] Figure 5 shows the results of particle size analysis of active fine particles according to one embodiment of the present invention.

[0032] Figure 6 is an SEM analysis photograph of active microparticles according to one embodiment of the present invention.

[0033] Figure 7 shows the experimental results for the release of semaglutide from active microparticles bound to semaglutide according to one embodiment of the present invention.

[0034] The present invention relates to a method for producing active microparticles according to the characteristics of a drug, comprising the steps of: dissolving a biodegradable polymer in an organic solvent to produce an oily solution; dissolving a surfactant in water to produce an aqueous solution; producing an emulsion using the oily solution and the aqueous solution; and freeze-drying the emulsion to produce active microparticles.

[0035] Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0036] WeGovi is a drug used for weight management and obesity treatment, and is a leading GLP-1 receptor agonist. WeGovi is administered as a subcutaneous injection once a week, and its effects can last as long as the drug remains in the body. With consistent use, weight loss typically lasts for several weeks to months. However, if the user discontinues use, the weight loss effect may gradually disappear.

[0037] As mentioned above, semaglutide is a GLP-1 (appetite-suppressing hormone) receptor agonist used to treat obesity and manage type 2 diabetes. However, currently commercialized products require users to take the medication once a week, consistently, and this consistent dosing can improve medication adherence. Furthermore, in situations requiring weight loss and blood sugar control, it's crucial to maintain a stable level of medication in the body. Semaglutide maximizes its effectiveness when blood concentrations remain stable. Therefore, users may feel more confident when they believe the medication will have a sustained effect, which can lead to psychological stability and continued motivation for treatment.

[0038] Consequently, in order to increase the effectiveness of treating diabetes or preventing and / or treating obesity by semaglutide, the lower the frequency of administration, the less burden there is on the user, and the more the therapeutic effect can be maintained.

[0039] However, the semaglutide mentioned above is currently only commercialized as a once-weekly product, and although a long-acting formulation that lasts for more than one month is under development, a representative technology for such a long-acting formulation is to manufacture particles using a biodegradable polymer. In the case of the formulation, the biodegradable polymer and semaglutide are evenly distributed within the particle, and as the biodegradable polymer decomposes in the body, semaglutide within the particle can be released into the body, thereby exhibiting a release effect of semaglutide for more than one month.

[0040] In order to manufacture a formulation that releases semaglutide for more than one month as described above, a manufacturing method such as a solvent evaporation method, a membrane method, or a microfluidic method must be used.

[0041] Solvent evaporation is one method for producing microparticles or nanoparticles, and is manufactured through the following steps. First, a polymer solution is prepared. This step involves dissolving a biodegradable polymer (e.g., polylactide (PLA) or polylactic acid-co-glycolic acid (PLGA)) in an organic solvent (e.g., acetone, chloroform, etc.). The polymer concentration and solvent selection are crucial in this step, as they can affect the size and structure of the final particles.

[0042] The second step is the emulsification step. This emulsification step involves mixing the dissolved polymer solution with water or another non-polar solution to facilitate the emulsification process. High-speed stirring or ultrasound can be used to ensure the two solutions are well mixed. This allows the polymer solution to be dispersed into small droplets.

[0043] The third step is the solvent evaporation step. This step involves the evaporation of the organic solvent from the emulsified mixture, causing the polymer to precipitate and form particles. Temperature, pressure, and ventilation rate are crucial in this step, and the evaporation rate can be controlled by these conditions. Once the solvent has completely evaporated, the polymer can aggregate to form particles.

[0044] The fourth step is particle collection and washing. The generated particles are collected through centrifugation or filtering and can be washed to remove any remaining solvent or impurities.

[0045] The fifth step is the drying step. Finally, the particles produced can be completely dried through freeze-drying or vacuum drying. This can increase particle stability and long-term storage potential. To apply the drug delivery system using the solvent evaporation method described above, semaglutide can be dissolved in the polymer solution during preparation.

[0046] In order to manufacture long-lasting micro-particles or nano-particles containing semaglutide using the solvent evaporation method described above, a mixer, an emulsifier, an evaporator, a centrifuge, a washing and drying device, a monitoring and control system, etc. are separately required.

[0047] In addition to the above solvent evaporation method, membrane method or microfluidic method also require separate equipment to carry out the corresponding process, such as solvent evaporation method.

[0048] Accordingly, in order to manufacture a long-acting formulation using semaglutide or a pharmaceutically acceptable salt thereof, a separate manufacturing process is required to carry out a specific process.

[0049] In addition to the aforementioned semaglutide, various drugs require development into long-acting formulations. Examples of drugs that could be developed into long-acting formulations include goserelin, leuprolide, progesterone, liraglutide, tirzepatide, ropivacaine, insulin, donepezil hydrochloride, rivastigmine, octreotide, rosuvastatin, dexmethylphenidate, and tamsulosin hydrochloride. Not limited to the above examples, any drug that requires a single injection to maintain its effect for more than one day, more than one week, or more than one month can be said to be a drug applicable to the present invention without limitation. However, as described above, in order to manufacture a long-acting formulation, a separate manufacturing process is required to carry out a specific process.

[0050] Accordingly, the present invention aims to provide active microparticles capable of producing a long-lasting formulation using only a stirrer, an incubator, and a freeze dryer without requiring any special equipment for a separate manufacturing process.

[0051] The above active microparticles can be combined with various drugs, such as the drugs described above, to form a long-acting formulation.

[0052] In addition, the surface charge, pore diameter, etc. of the above active microparticles can be adjusted according to the type of drug to be bound, thereby facilitating binding, and the type and density of the polymer, particle size, etc. can be adjusted to enable the drug to be continuously released in the body for a desired period of time.

[0053] The active microparticles of the present invention described above may include a step of preparing an oily solution by dissolving a biodegradable polymer in an organic solvent; a step of preparing an aqueous solution by dissolving a surfactant in water; a step of preparing an emulsion using the oily solution and the aqueous solution; and a step of preparing active microparticles by freeze-drying the emulsion.

[0054] The biodegradable polymer may be selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid and combinations thereof, and preferably may be polylactide-co-glycolide (PLGA).

[0055] In addition, the polylactide-co-glycolide may have a monomer ratio of lactide:glycolide of 100:0, 75:35, or 50:50. In addition, the terminal group of the polylactide-co-glycolide may be a carboxyl group or an ester group, and the molecular weight may be selected from a range of 2K to 200K Da.

[0056] The above biodegradable polymer can be selected according to the type of drug as described above, but when the terminal group of the polylactide-co-glycolide is a carboxyl group or an ester group, a chemical bond can occur with the drug through electrostatic attraction.

[0057] The above biodegradable polymer can be dissolved in an organic solvent to prepare an oily solution. The oily solution is at least one selected from the group consisting of dichloromethane, ethyl acetate, benzyl alcohol, acetone, methanol, ethanol, chloroform, chloroethane, dichloroethane, trichloroethane, and mixtures thereof, preferably dichloromethane, but not limited to the example, and any organic solvent capable of dissolving the biodegradable polymer, and not limited to the above examples, can be used as long as it is an organic solvent that can be easily selected by a person skilled in the art.

[0058] The surfactant contained in the above-mentioned aqueous solution may be contained in an amount of 0.1 to 5.0 wt%, 0.2 to 1 wt%, 0.25 to 0.75 wt%, or 0.5 wt%. The remainder is water.

[0059] The above surfactants include methylcellulose, polyvinylpyrrolidone, lecithin, gelatin, polyvinyl alcohol, sorbitan monooleate (e.g., Span 80). TM 80), polyoxyethylene sorbitan fatty acid esters (e.g., Tween 80) TM80) etc.), polyoxyethylene castor oil derivatives, sodium lauryl sulfate, sodium stearate, ester amine, linear diamine, patty amine and mixtures thereof, preferably polyvinyl alcohol, but not limited to the above examples, any surfactant that can be prepared into a completely spherical emulsion can be used.

[0060] The step of preparing an emulsion using the above oily solution and aqueous solution can be prepared by using a membrane, a homogenizer, or a microfluidic method, and is not limited to the above examples. An oily solution containing a biodegradable polymer and an aqueous solution containing a surfactant are used, and any preparation method capable of forming an emulsion within the interface of the oily solution and the aqueous solution is not limited and can be used. In the present invention, the emulsion was prepared using a membrane method.

[0061] In order to prepare an emulsion by the above membrane method, the oil phase solution can be injected at an injection rate of 0.1 to 10.0 mL / min., and the water phase solution can be injected at an injection rate of 10.0 to 150.0 mL / min., each using a pump.

[0062] Forming an emulsion by the above membrane method can be applied equally to all the manufacturing processes described below. The emulsion manufactured by the above membrane method can be solidified to remove the organic solvent and then subjected to a freeze-drying process. To minimize the residual organic solvent, a vacuum drying step is additionally performed. More specifically, a solidification step is performed at 20°C to 30°C for 20 to 30 hours, after which the residual organic solvent is removed and freeze-dried. After the solidification step, vacuum drying can be additionally performed at 35°C to 50°C for 40 to 55 hours to minimize the residual organic solvent.

[0063] The above active fine particles may have an average diameter (D50) of 10 µm to 100 µm, 10 µm to 95 µm, 10 µm to 90 µm, or 10 µm to 85 µm.

[0064] The above active particles have a specific surface area of ​​1 m 2 / g to 15 m 2 / g, 2m 2 / g to 14 m 2 / g. The above specific surface area is the total surface area of ​​the active microparticles, and may vary depending on whether pores are formed, the shape of the pores, the total number of pores, etc. When semaglutide or a pharmaceutically acceptable salt thereof is combined using active microparticles having a specific surface area within the above range, it is possible to manufacture a long-acting formulation that continuously releases semaglutide or a pharmaceutically acceptable salt thereof in the body for more than one month.

[0065] The above-described manufacturing method is for a case where the terminal group of the biodegradable polymer is a carboxyl group or an ester group, and the terminal group of the biodegradable polymer as described above may be for inducing a chemical bond with a drug.

[0066] According to another embodiment of the present invention, the active microparticles may be characterized by controlling the size of internal pores or controlling the charge of the active microparticles in addition to the terminal groups of the biodegradable polymers described above to facilitate binding to a drug.

[0067] Specifically, the conventional long-acting formulation including a biodegradable polymer is manufactured by dissolving the biodegradable polymer and the drug in an organic solvent and then mixing it with an aqueous solution, and the microspheres manufactured by the above manufacturing method are not formed by chemical bonding between the biodegradable polymer and the drug, as the biodegradable polymer and the drug are uniformly mixed. On the other hand, the active microparticles of the present invention are characterized in that the drug is not included in the manufacturing process of the microparticles, and the active microparticles are first manufactured and then bound to the drug using the manufacturing process described below.

[0068] In this regard, the aforementioned drugs have various sizes, and depending on the size of the drug, differences in binding to the active microparticles may occur. To address this issue, the present invention utilizes an activator to control the size of the internal pores when manufacturing the active microparticles.

[0069] Specifically, the activator may be a channel former. The channel former may be selected from the group consisting of polyvinyl alcohol (PVA), ammonium bicarbonate (ABC), bovine serum albumin (BSA), trehalose, ethanol, pluronic, calcium chloride (CaCl2), surfactants, and mixtures thereof, but is not limited to the above examples and any agent capable of forming internal pores of the active microparticles may be used without limitation.

[0070] The above surfactants include methylcellulose, polyvinylpyrrolidone, lecithin, gelatin, polyvinyl alcohol, sorbitan monooleate (e.g., Span 80). TM 80), polyoxyethylene sorbitan fatty acid esters (e.g., Tween 80) TM 80) etc.), polyoxyethylene castor oil derivatives, sodium lauryl sulfate, sodium stearate, ester amine, linear diamine, patty amine and mixtures thereof, but is not limited to the above examples and any one that can be used for pore formation can be used without limitation.

[0071] The above channel forming agent can be included in the production of active microparticles as described above, and can control the internal pore size of the microparticles, etc., thereby facilitating binding to a drug.

[0072] Another activator may include a binding activator. The binding activator may facilitate chemical bonding between the active microparticle and the drug, such as ionic interaction, hydrophobic interaction, π-π interaction, hydrogen bonding, or protein binding.

[0073] The above binding activator may be selected from the group consisting of polyethyleneimine (PEI), protamine sulfate, bovine serum albumin (BSA), poly-L-lysine, calcium chloride (CaCl2), and mixtures thereof.

[0074] The above activator may be included as a channel forming agent and / or a binding activator, and depending on the type of drug, only one type may be selected and included, or both types may be included.

[0075] The above activator can be classified as a water-soluble activator based on its solubility properties. If it is not a water-soluble activator, the activator can be mixed with the biodegradable polymer in the step of preparing an oily solution by dissolving the biodegradable polymer in an organic solvent. The steps for preparing the active microparticles thereafter are identical to those described above.

[0076] On the other hand, in the case of a water-soluble activator, as described above, it has the characteristic of not being dissolved in organic solvents, and thus cannot be directly dissolved in an organic solvent together with a biodegradable polymer. For this reason, in the case of a water-soluble activator, a solution containing the activator can be prepared by dissolving it in distilled water, and then mixing it with an oily solution in which the biodegradable polymer is dissolved in an organic solvent to form a first emulsion. In order to prepare a solution containing the activator, a solution containing the activator can be prepared by mixing 10 mg to 500 mg of the water-soluble activator with 1 ml of distilled water.

[0077] The step of forming a dispersion solution by mixing a solution containing the above-mentioned activator and an oily solution may be performed by mixing the first aqueous solution and the oily solution and stirring them using a homogenizer at 10,000 to 15,000 rpm for 30 to 120 seconds to form a dispersion solution.

[0078] Thereafter, the step of preparing an aqueous solution by dissolving the surfactant in water as described above may be preparing an aqueous solution by dissolving 0.5 wt% of the surfactant in water.

[0079] The emulsion can be formed using a step of preparing an emulsion using the above-described dispersion solution and aqueous solution, a stirring method using the above-described homogenizer, a membrane method, a microfluidic method, etc.

[0080] In the present invention, a membrane having a pore size of 5 μm to 10 μm was used. Through the dispersion solution inlet of the membrane, the dispersion solution was injected at an injection rate of 0.5 ml / min to 2.0 ml / min, and the aqueous solution was injected at an injection rate of 20 ml / min to 40 ml / min, through each connected pump, to form an emulsion.

[0081] The above emulsion can be solidified at 20°C to 30°C for 20 to 30 hours, after which the remaining organic solvent is removed and freeze-dried.

[0082] After the above-mentioned drying step, vacuum drying may be additionally performed at 35°C to 50°C for 40 to 55 hours to minimize residual organic solvent.

[0083] Through the above process, active microparticles can be manufactured. The active microparticles, as described above, may have an average diameter (D50) of 10 µm to 100 µm, 10 µm to 95 µm, 10 µm to 90 µm, or 10 µm to 85 µm. In addition, the specific surface area determined by the N2 adsorption and desorption isotherm at 196°C may be 1 m 2 / g to 15 m 2 / g, 2m 2 / g to 14 m 2 / g. In addition, the total pore volume determined by the N2 adsorption and desorption isotherm at 196℃ is 0.005 cm 3 / g to 0.020 cm 3 / g, 0.006 cm 3 / g to 0.019 cm 3 / g, 0.007 cm 3 / g to 0.019 cm 3 / g, 0.008 cm 3 / g to 0.019 cm 3 / g, 0.009 cm 3 / g to 0.019 cm 3 / g. In addition, the average pore diameter determined by the N2 adsorption and desorption isotherm at 196℃ may be 1.0 nm to 10.0 nm, 1.5 nm to 9.5 nm, 2.0 nm to 9.0 nm, 2.0 nm to 8.5 nm, or 2.0 nm to 8.0 nm. By the above characteristics, it is possible to more easily bind the drug to the internal pores, thereby enabling the drug to be provided in a long-lasting formulation in a bound state by utilizing active microparticles.

[0084] According to another embodiment of the present invention, the active microparticles may be manufactured using the manufacturing method described above. Furthermore, the active microparticles may be provided in a state in which they are bound to a drug, as described below, and the active microparticles bound to the drug may be provided as an injectable composition.

[0085] In the present invention, the composition may include active microparticles containing a therapeutically effective amount of a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In addition, the composition may include a drug selected from the group consisting of goserelin, progesterone, liraglutide, tirzepatide, ropivacaine, insulin, donepezil hydrochloride, rivastigmine, octreotide, rosuvastatin, dexmethylphenidate, tamsulosin hydrochloride, and the like, or a pharmaceutically acceptable salt thereof, rather than a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof. The above "pharmaceutically acceptable carrier" is a substance that can be added to an active ingredient to help formulate or stabilize the preparation, and does not cause significant harmful toxic effects to the patient.

[0086] The above carrier refers to a carrier or diluent that does not irritate the patient and does not inhibit biological activity and properties. In a composition formulated as a liquid solution, acceptable pharmaceutical carriers include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these components. If necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

[0087] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the preparation of extemporaneous sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions are preferably formulated for injection. The compositions may be formulated as solutions, microemulsions, liposomes, or other tailored structures suitable for high drug concentrations. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. In some cases, isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride, may be included in the composition.

[0088] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterile microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the other required ingredients from those enumerated above. For sterile powders for the preparation of sterile injectable solutions, some preparation methods are vacuum drying and freeze-drying (lyophilization), which produce a powder of the active ingredient and any additional desired ingredient from a previously sterile-filtered solution thereof.

[0089] Specifically, the composition may contain formulation materials to modify, maintain, or preserve the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption, or permeation of the composition. Suitable formulation materials include amino acids (e.g., glycine, glutamine, asparagine, arginine, or lysine), antimicrobial agents, antioxidants (e.g., ascorbic acid, sodium sulfite, or sodium bisulfite), buffers (e.g., borates, bicarbonates, Tris-HCl, citrates, phosphates, or other organic acids), bulking agents (e.g., mannitol or glycine), chelating agents (e.g., ethylenediamine tetraacetic acid (EDTA)), complexing agents (e.g., caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin), fillers, monosaccharides, disaccharides, and other carbohydrates (e.g., glucose, mannose, or dextrins), proteins (e.g., serum albumin, gelatin, or immunoglobulins), coloring agents, flavoring agents, and diluents, emulsifiers, hydrophilic agents. Polymers (e.g. polyvinylpyrrolidone), low molecular weight polypeptides, salt-forming counterions (e.g. sodium), preservatives (e.g. benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide), solvents (e.g. glycerin, propylene glycol or polyethylene glycol), sugar alcohols (e.g. mannitol or sorbitol), suspending agents, surfactants or wetting agents (e.g. pluronics; PEG; sorbitan esters; polysorbates, such as polysorbate 20 or polysorbate 80; Triton; tromethamine; lecithin; cholesterol or tyloxapal), stability enhancers (e.g. sucrose or sorbitol), tonicity enhancers (e.g. alkali metal halides; preferably sodium chloride or potassium chloride; or mannitol sorbitol), delivery vehicles, diluents, excipients and / or pharmaceutical adjuvants, including but not limited to.

[0090] In the present invention, the frequency of administration of the composition will vary depending on the pharmacokinetic parameters of the drug-containing active microparticles in the formulation used. Typically, the clinician will administer the composition until the desired effect is achieved. Thus, the composition may be administered as a single dose, as two or more doses spaced apart (which may or may not contain the same amount of drug-containing active microparticles), or as a continuous infusion via an implantable device or catheter. Further refinement of the appropriate dosage is routinely accomplished by those skilled in the art and falls within the scope of their routine practice.

[0091] The unit dosage for humans is 0.01 μg / kg to 100 mg / kg, specifically 1 μg / kg to 30 mg / kg. Although the above amount is the optimal amount, it may vary depending on the disease being treated and the presence or absence of side effects, and the optimal dosage can be determined through routine experiments.

[0092] A method for producing a long-acting formulation containing a drug using active microparticles according to another embodiment of the present invention may include the steps of: adding the active microparticles of the present invention to a buffer solution and stirring to produce a suspension solution containing the active microparticles; and adding a drug to the suspension solution containing the active microparticles and stirring to bind the drug to the active microparticles.

[0093] The above active microparticles are manufactured using a biodegradable polymer as described above, and the manufactured active microparticles are manufactured as porous microparticles using only a biodegradable polymer without any separate drug bound to them.

[0094] The above active microparticles can be placed in a buffer solution and stirred to prepare a suspension solution.

[0095] The above suspension solution typically refers to a state in which fine particles are uniformly dispersed within the solution. The fine particles within the solution may be uniformly dispersed without being dissolved. However, to prepare the above suspension solution, an activator may be additionally included depending on the potential state of the active fine particles.

[0096] When the zeta potential of the above-mentioned active microparticles is negative, the zeta potential can be changed to positive by including an activator.

[0097] As described above, the activator capable of changing the zeta potential of the active microparticles to a positive charge may be selected from the group consisting of protamine sulfate, polyethyleneimine (PEI), bovine serum albumin (BSA), poly-L-lysine, calcium chloride (CaCl2), and mixtures thereof, preferably protamine sulfate, but not limited to the above examples. In order to prepare the suspension solution, the active microparticles and the activator may be included in a weight ratio of 1:0.5 to 1:2.0.

[0098] For preparing the above suspension solution, the buffer solution may be selected from the group consisting of HEPES (4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), Tris (tris(hydroxymethyl)aminomethane) and mixtures thereof, preferably a HEPES buffer solution, but is not limited to the above examples, and any buffer solution that can be prepared as a suspension solution by mixing active microparticles may be used without limitation.

[0099] After preparing a suspension solution as described above, a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof is added and stirred to bind the GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof to the active microparticles.

[0100] Alternatively, a drug selected from among goserelin, leuprolide, progesterone, liraglutide, tirzepatide, ropivacaine, insulin, donepezil hydrochloride, rivastigmine, octreotide, rosuvastatin, dexmethylphenidate, tamsulosin hydrochloride, etc., other than a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof, may be added to the suspension solution and stirred to bind to the active microparticles.

[0101] The drug (API) or a pharmaceutically acceptable salt thereof mixed in the above suspension solution may be included in a weight ratio of 1:5 to 1:15, a weight ratio of 1:6 to 1:14, a weight ratio of 1:7 to 1:13, a weight ratio of 1:8 to 1:12, a weight ratio of 1:8 to 1:11, or a weight ratio of 1:8 to 1:10, relative to the active microparticles, and when mixed within the above range, the drug (API) or a pharmaceutically acceptable salt thereof may be chemically bonded to the active microparticles.

[0102] However, the active microparticles, in which the drug or its pharmaceutically acceptable salt is bound as described above, may have a large number of pores formed on their surface. In this state, as described above, moisture may penetrate the surface pores of the active microparticles, causing rapid decomposition of the microparticles due to hydrolysis. Due to this problem, the drug or its pharmaceutically acceptable salt contained in the active microparticles may be released quickly, resulting in a problem in which the drug or its pharmaceutically acceptable salt may not be released for more than one month.

[0103] To prevent such problems, after the step of binding a drug or a pharmaceutically acceptable salt thereof to the active microparticles, an additional step of incubating the active microparticles at a temperature higher than the Tg temperature of the biodegradable polymer included in the active microparticles for 1 to 5 hours may be performed.

[0104] The above-mentioned Tg temperature condition may be 40°C or higher. The Tg temperature of the biodegradable polymer, when the biodegradable polymer is PLGA, may be more specifically 40°C to 50°C, 40°C to 49°C, 40°C to 48°C, 40°C to 47°C, 40°C to 46°C, 40°C to 45°C, 40°C to 44°C, 40°C to 43°C, or 40°C to 42°C. When incubated for 1 hour to 5 hours, 1 hour to 4 hours, 1 hour to 3 hours, or 1 hour to 2 hours under the above-mentioned temperature conditions, the pores formed on the outside of the active microparticles may be closed by self-healing.

[0105] As described above, when the external pores of the active microparticles are closed, even if the active microparticles are administered into the body, moisture does not penetrate, preventing rapid decomposition.

[0106] As described above, the step of manufacturing active microparticles containing various drugs or pharmaceutically acceptable salts thereof of the present invention is a process for chemically bonding various drugs using already manufactured active microparticles, and does not require a separate device for performing the process.

[0107] Ultimately, it will be possible to manufacture long-acting formulations containing a variety of drugs using existing drug manufacturing facilities.

[0108] This means that it is possible to produce long-acting formulations containing various drugs, overcoming the problem that manufacturing was only possible in places where there was equipment to manufacture specific long-acting formulations.

[0109] Analysis method

[0110] Particle size analysis using a laser particle size analyzer

[0111] 50 mg of fine particles were suspended in purified water (1.0 mL), mixed with a vortex mixer for 20 seconds, and then dispersed in an ultrasonic generator for 1 minute. The microsphere dispersion was analyzed using a Beckman Coulter laser particle size analyzer (LS 13 320 XR particle size analyzer, Beckman Coulter, USA) in a ULM (Universal Liquid Module) in the range of 0.01–3500 μm.

[0112] Morphological analysis using scanning electron microscopy

[0113] 5 mg of microspheres were placed on an aluminum stub with carbon tape attached and coated with platinum. The aluminum stub was mounted in a field-emission scanning electron microscope (FE-SEM, JSM-6700F, JEOL, Japan), and the morphological characteristics of the microspheres were observed at an acceleration voltage of 15 kV. The electron microscope images of the microspheres were confirmed.

[0114] Specific surface area analysis

[0115] 0.25 to 1.0 g of fine particles were vacuum-dried at 40°C for 24 hours. The specific surface area of ​​the dried fine particles was determined by N2 adsorption / desorption isotherm at -196°C using Micromeritics (TriStar II 3020 Version 3.02 / Micromeritics instrument corporation).

[0116] Encapsulation rate analysis

[0117] After completely dissolving the fine particles (5 mg) in ACN, they were diluted with the mobile phase. 50 μL of the diluted solution was injected into HPLC and measured at a detection wavelength appropriate for the characteristics of each drug.

[0118] Encapsulation Efficiency(%)=(weight of the drug in microshpere / weight of feeding drug×100)

[0119] Manufacturing example

[0120] Manufacturing of active microparticles

[0121] Example 1

[0122] 3.0 g of PLGA(100 / 0)-COOH polymer with a molecular weight of 12 KDa was dissolved in 10 g of methylene chloride to prepare an oil phase (DP, Disperse Phase) solution. 0.5% PVA (Polyvinyl alcohol) was dissolved in purified water to prepare an aqueous phase (CP, Continuous Phase) solution. The DP and CP were injected at an injection rate of 1.0 mL / min and 30.0 mL / min, respectively, using a pump so that an emulsion was formed simultaneously with the injection. The emulsion was prepared using a membrane with a pore size of 5 μm. The organic solvent was removed through a solidification step at 25°C for 24 hours, and a freeze-drying process was performed.

[0123] Example 2

[0124] It was manufactured using PLGA(100 / 0)-COOH polymer with a molecular weight of 30 KDa, and the other manufacturing processes are the same as in Example 1.

[0125] Example 3

[0126] It was manufactured using PLGA (50 / 50)-COOH polymer with a molecular weight of 12 KDa, and the other manufacturing processes are the same as in Example 1.

[0127] Example 4

[0128] It was manufactured using PLGA (50 / 50)-COOH polymer with a molecular weight of 30 KDa, and the other manufacturing processes are the same as in Example 1.

[0129] Example 5

[0130] It was manufactured using PLGA (75 / 25)-COOH polymer with a molecular weight of 12 KDa, and the other manufacturing processes are the same as in Example 1.

[0131] Example 6

[0132] It was manufactured using PLGA (75 / 25)-COOH polymer with a molecular weight of 30 KDa, and the other manufacturing processes are the same as in Example 1.

[0133] Example 7

[0134] It was manufactured using PLGA (50 / 50)-COOR polymer with a molecular weight of 12 KDa, and the other manufacturing processes are the same as in Example 1.

[0135] Example 8

[0136] It was manufactured using PLGA (75 / 25)-COOR polymer with a molecular weight of 12 KDa, and the other manufacturing processes are the same as in Example 1.

[0137] The analysis results for the active microparticles manufactured according to Examples 1 to 8 are as shown in Table 1 below:

[0138] No. Polymer type Size (D10 / D50 / D90, ㎛) SEM Image Example 1202HL / G=100 / 0F-COOHMw=10K~15K10.8 / 19.5 / 30.3 Example 2203HL / G=100 / 0F-COOHMw=20K~30K12.1 / 20.7 / 30.6 Example 3502HL / G=50 / 50F-COOHMw=10K~15K9.5 / 17.0 / 26.0 Example 4503HL / G=50 / 50F-COOHMw=20K~30K15.7 / 25.3 / 43.2 Example 5752HL / G=75 / 25F-COOHMw=10K~15K12.1 / 19.7 / 29.0 Example 6753HL / G=75 / 25F-COOHMw=20K~30K19.1 / 28.1 / 42.4 Example 7502SL / G=50 / 50F-COORMw=10K~15K18.5 / 28.2 / 39.8 Example 8752SL / G=75 / 25F-COORMw=10K~15K19.1 / 29.2 / 42.3

[0139] Preparation of active microparticles using channel forming agents

[0140] Example 9

[0141] A 1.0% aqueous solution was prepared by dissolving polyvinyl alcohol (PVA), a water-soluble channel forming agent, in distilled water. 3.0 g of PLGA (75 / 25)-COOH polymer with a molecular weight of 12 KDa was dissolved in 10 g of methylene chloride to prepare an oily solution, and the aqueous solution containing PVA and the oily solution were mixed to prepare a dispersed solution (DP, Disperse Phase). The weight ratio of PVA to the weight of PLGA in the dispersed solution is shown in Table 2 below. A continuous phase solution (CP, Continuous Phase) was prepared by dissolving 0.5% PVA (Polyvinyl alcohol) in purified water. The DP and CP were injected using a pump at an injection rate of 1.0 mL / min and 30.0 mL / min, respectively, so that an emulsion was formed simultaneously with the injection. The emulsion was prepared using a membrane with a pore size of 5 μm. After a 24-hour solidification step at 25°C, the organic solvent was removed and a freeze-drying process was performed.

[0142] Example 10

[0143] ABC, a water-soluble channel forming agent, was dissolved in distilled water to prepare a 3.0% aqueous solution, and the other manufacturing processes were the same as in Example 9.

[0144] Example 11

[0145] BSA, a water-soluble channel forming agent, was dissolved in distilled water to prepare a 30.0% aqueous solution, and the other manufacturing processes were the same as in Example 9.

[0146] Example 12

[0147] Trehalose, a water-soluble channel forming agent, was dissolved in distilled water to prepare a 3.0% aqueous solution, and the other manufacturing processes were the same as in Example 9.

[0148] Example 13

[0149] Ethanol, a water-soluble channel forming agent, was dissolved in distilled water to prepare a 50.0% aqueous solution, and the other manufacturing processes were the same as in Example 9.

[0150] Example 14

[0151] Pluronic 188, a water-soluble channel forming agent, was dissolved in distilled water to prepare a 6.0% aqueous solution, and the other manufacturing processes were the same as in Example 9.

[0152] Example 15

[0153] Surfactant Span 80, a water-soluble channel forming agent, was dissolved in distilled water to prepare a 15.0% aqueous solution, and the other manufacturing processes were the same as in Example 9.

[0154] The analysis results for the active microparticles manufactured by Examples 9 to 15 are as shown in Table 2 below:

[0155] No. Channel forming agent type Weight ratio of channel forming agent to polymer Size (D10 / D50 / D90, ㎛) SEM Image Example 9 PVA 0.33 wt% 16.1 / 26.8 / 46.3 Example 10ABC1 wt% 16.6 / 26.3 / 38.4 Example 11BSA10 wt% 31.4 / 42.4 / 56.2 Example 12Trehalose 1 wt% 13.8 / 22.6 / 33.0 Example 13 Ethanol 16.67 wt% 18.1 / 32.8 / 51.0 Example 14Pluronic2 wt% 12.1 / 19.7 / 29.0 Example 15Surfactant 5 wt% 26.0 / 39.2 / 55.9

[0156] Preparation of active microparticles using binding activators

[0157] Example 16

[0158] PEI, a water-soluble activating agent, was dissolved in distilled water to prepare a 7.5% aqueous solution. 3.0 g of PLGA (75 / 25)-COOH polymer with a molecular weight of 12 KDa was dissolved in 10 g of methylene chloride to prepare an oily solution. The weight ratio of PEI to the weight of PLGA in the oily solution is shown in Table 3 below. 0.5% PVA (polyvinyl alcohol) was dissolved in purified water to prepare a continuous phase (CP) solution. The DP was injected at an injection rate of 1.0 mL / min and the CP was injected at an injection rate of 30.0 mL / min using a pump so that an emulsion was formed simultaneously with the injection. The emulsion was prepared using a membrane with a pore size of 5 μm. The organic solvent was removed through a solidification step at 25°C for 24 hours, and a freeze-drying process was performed.

[0159] Example 17

[0160] BSA, a water-soluble activating agent, was dissolved in distilled water to prepare a 30.0% aqueous solution, and the other manufacturing processes were the same as in Example 16.

[0161] Example 18

[0162] Protamine sulfate, a water-soluble activating agent, was dissolved in distilled water to prepare a 3.0% aqueous solution, and the other manufacturing processes were the same as in Example 16.

[0163] Example 19

[0164] A 3.0% aqueous solution was prepared by dissolving CaCl2, a water-soluble activating agent, in distilled water, and the other manufacturing processes were the same as in Example 16.

[0165] The analysis results for the active microparticles manufactured by Examples 16 to 19 are as shown in Table 3 below:

[0166] No. Activator type Weight ratio of channel forming agent to polymer Size (D10 / D50 / D90, ㎛) SEM Image Example 16 PEI 2.5 wt% 11.7 / 27.1 / 43.6 Example 17BSA10 wt% 32.1 / 46.7 / 73.2 Example 18 Protamine sulfate 1 wt% 34.0 / 50.2 / 72.4 Example 19CaCl21 wt%18.5 / 32.3 / 52.9

[0167] Preparation of active microparticles using channel forming agents and binding activators

[0168] Example 20

[0169] ABC and BSA were dissolved in distilled water to prepare 1.0% aqueous solutions, respectively. 3.0 g of PLGA (75 / 25)-COOH polymer with a molecular weight of 12 KDa was dissolved in 10 g of methylene chloride to prepare an oily solution, and the aqueous solution containing ABC and the aqueous solution containing BSA prepared above were mixed with the oily solution in a weight ratio of 1:1 to prepare a dispersed solution (DP, Disperse Phase). ABC and BSA in the dispersed solution were mixed so that each was 1 wt% based on the weight of the PLGA. 0.5% PVA (Polyvinyl alcohol) was dissolved in purified water to prepare a secondary aqueous phase (CP, Continuous Phase). The DP was injected at an injection rate of 1.0 mL / min. and the CP was injected at an injection rate of 30.0 mL / min. using a pump so that an emulsion was formed simultaneously with the injection. The emulsion was prepared using a membrane with a pore size of 5 μm. After a 24-hour solidification step at 25°C, the organic solvent was removed and the freeze-drying process was performed.

[0170] Example 21

[0171] For the composite active microparticles, PVA and CaCl2 were dissolved in distilled water to prepare 1.0% and 3.0% aqueous solutions, respectively, and the other manufacturing processes were the same as in Example 20.

[0172] Example 22

[0173] For the composite active microparticles, PVA and ABC were dissolved in distilled water to prepare 10.0% and 3.0% aqueous solutions, respectively, and the other manufacturing processes were the same as in Example 20.

[0174] Example 23

[0175] For the composite active microparticles, BSA and CaCl2 were dissolved in distilled water to prepare a 3.0% aqueous solution, and the other manufacturing processes were the same as in Example 20.

[0176] Example 24

[0177] For the composite active microparticles, EtOH and CaCl2 were dissolved in distilled water to prepare 50.0% and 3.0% aqueous solutions, respectively, and the other manufacturing processes were the same as in Example 20.

[0178] Example 25

[0179] For the composite active microparticles, ABC and Span 80 were dissolved in distilled water to prepare 3.0% and 15.0% aqueous solutions, respectively, and the other manufacturing processes were the same as in Example 20.

[0180] Example 26

[0181] For the composite active microparticles, ABC and Tween 20 were dissolved in distilled water to prepare 3.0% and 15.0% aqueous solutions, respectively, and the other manufacturing processes were the same as in Example 20.

[0182] Example 27

[0183] For the composite active microparticles, PEI and CaCl2 were dissolved in distilled water to prepare 7.5% and 3.0% aqueous solutions, respectively, and the other manufacturing processes were the same as in Example 20.

[0184] The analysis results for the active microparticles manufactured by Examples 20 to 27 are as shown in Table 4 below:

[0185] No. Weight ratio of channel forming agent to polymer type Size (D10 / D50 / D90, ㎛) SEM Image Example 20 ABC + BSAABC 1 wt% + BSA 1 wt% 17.2 / 27.9 / 40.6 Example 21PVA + CaCl2PVA 0.33 wt% + CaCl2 1 wt%20.4 / 30.8 / 43.5 Example 22PVA + ABCPVA 3.33 wt% + ABC 1 wt% 21.5 / 31.3 / 43.5 Example 23 BSA + CaCl2 BSA 1 wt% + CaCl2 1 wt% 14.6 / 58.0 / 86.2 Example 24 EtOH + CaCl2 EtOH 16.67 wt% + CaCl2 1 wt% 18.1 / 32.8 / 51.0 Example 25ABC + Span80ABC 1 wt% + Span80 5 wt% 18.9 / 29.9 / 43.8 Example 26ABC + Tween20ABC 1 wt% + Tween20 5 wt% 11.6 / 20.4 / 30.3 Example 27PEI + CaCl2PEI 2.5 wt% + CaCl2 1 wt%50.9 / 82.2 / 129.7

[0186]

[0187] Preparation of sustained-release microparticles containing goserelin

[0188] (1) Manufacturing of active microparticles

[0189] The first phase solution, a 3.0% CaCl2 aqueous solution, was prepared by dissolving CaCl2, a water-soluble channel-forming agent, in distilled water. An oily solution was prepared by dissolving 3.0 g of PLGA 5050 polymer in 10 g of methylene chloride. The first phase solution and the oily solution were mixed to prepare a dispersion solution. A 0.5% PVA aqueous solution was prepared as the second phase solution.

[0190] The above dispersion solution was injected using a pump at an injection rate of 1.0 mL / min., and the second aqueous solution was injected using a pump at an injection rate of 30.0 mL / min., so that an emulsion was formed simultaneously with the injection. The emulsion was formed using a membrane (PHS membrane) with a pore diameter of 5 μm. The formed emulsion went through a solidification step at 25°C for 24 hours, removed the organic solvent, and then proceeded to a freeze-drying process.

[0191] (2) Goserelin loading method

[0192] The above-mentioned active microparticles (270 mg) and goserelin (30 mg) were suspended in HEPES buffer pH 7.4 (3.0 mL). The mixture was stirred with a rotary mixer in an incubator at 37°C for 24 hours. An additional 2-hour incubation was performed at 40-42°C to control the initial drug release. Upon completion of the incubation, the mixture was centrifuged at 7012 g for 5 minutes to remove the supernatant and washed three times with 3.0 mL of fresh purified water. The mixture was lyophilized to obtain microparticles loaded with goserelin.

[0193] The results of particle size analysis of fine particles bound with goserelin using the above-mentioned active fine particles are as shown in Fig. 1. According to the results of particle size analysis, D10 / D50 / D90 were 22.6 ㎛, 34.8 ㎛, and 51.4 ㎛, respectively, and the span value was 0.8, confirming that uniform particles were formed.

[0194] In addition, the SEM image measurement results are as shown in Fig. 2. In addition, the specific surface area determined by N- adsorption / desorption isotherm at -196°C using Micromeritics (TriStar II 3020 Version 3.02 / Micromeritics instrument corporation) is 4.3200 m2 / g, and the pore volume (Pore Volume, P / P0=0.9918) is 0.0091 cm 3 / g, and the pore size (adsorption average pore diameter) is 2.3397 nm.

[0195] Additionally, the encapsulation rate for goserelin was confirmed to be 87.6%.

[0196]

[0197] Preparation of sustained-release microparticles containing semaglutide

[0198] (1) Manufacturing of active microparticles

[0199] An oily solution was prepared by dissolving 3.0 g of PLGA 7525 polymer in 10 g of methylene chloride. Aqueous solutions of ABC and BSA were each prepared by dissolving them in distilled water. A dispersion solution was prepared by mixing the aqueous solution containing ABC and the aqueous solution containing BSA with the oily solution. A 0.5% aqueous PVA solution was used as the aqueous solution.

[0200] The above dispersion solution was injected at an injection rate of 1.0 mL / min., and the above aqueous solution was injected at an injection rate of 30.0 mL / min., respectively, using a pump to form an emulsion. The emulsion was formed using a membrane contact (PHS membrane) with a pore diameter of 5 μm. The formed emulsion went through a solidification step at 25°C for 24 hours, removed the organic solvent, and then proceeded to a freeze-drying process.

[0201] (2) Semaglutide loading method

[0202] The above-mentioned active microparticles (270 mg) and semaglutide (30 mg) were suspended in HEPES buffer pH 7.4 (3.0 mL). The mixture was stirred with a rotary mixer in an incubator at 37°C for 24 hours. An additional 2-hour incubation was performed at 40-42°C to control the initial drug release. Upon completion of the incubation, the mixture was centrifuged at 7012 g for 5 minutes to remove the supernatant and washed three times with 3.0 mL of fresh purified water. The mixture was lyophilized to obtain microparticles loaded with semaglutide.

[0203] The particle size analysis results of the microparticles in which semaglutide is bound using the above-mentioned active microparticles are as shown in Fig. 3. According to the particle size analysis results, D10 / D50 / D90 are 16.3 ㎛, 25.5 ㎛, and 36.6 ㎛, respectively, and the span value is 0.8, confirming that uniform particles are formed.

[0204] In addition, the SEM image measurement results are as shown in Fig. 4. In addition, the specific surface area determined by N- adsorption / desorption isotherm at -196°C using Micromeritics (TriStar II 3020 Version 3.02 / Micromeritics instrument corporation) was 10.0654 m 2 / g, and the pore volume (Pore Volume, P / P0=0.9914) is 0.0185 cm 3 / g, and the pore size (adsorption average pore diameter) is 7.3544 nm.

[0205] Additionally, the encapsulation rate for semaglutide was confirmed to be 77.0%.

[0206]

[0207] Preparation of sustained-release microparticles containing ropivacaine

[0208] (1) Manufacturing of active microparticles

[0209] An oily solution was prepared by dissolving 3.0 g of PLGA 5050 polymer in 10 g of methylene chloride. Aqueous solutions of ABC and BSA were each prepared by dissolving them in distilled water. A dispersion solution was prepared by mixing the aqueous solution containing ABC and the aqueous solution containing BSA with the oily solution. A 0.5% aqueous PVA solution was used as the aqueous solution.

[0210] The above dispersion solution was injected at an injection rate of 1.0 mL / min., and the above aqueous solution was injected at an injection rate of 30.0 mL / min., respectively, using a pump to form an emulsion. The emulsion was formed using a membrane contact (PHS membrane) with a pore diameter of 5 μm. The formed emulsion went through a solidification step at 25°C for 24 hours, removed the organic solvent, and then proceeded to a freeze-drying process.

[0211] (2) Ropivacaine loading method

[0212] The above-mentioned active microparticles (240 mg) and ropivacaine (60 mg) were suspended in HEPES buffer pH 7.4 (3.0 mL). The mixture was stirred with a rotary mixer in an incubator at 37°C for 24 hours. An additional 2-hour incubation was performed at 40-42°C to control the initial drug release. Upon completion of the incubation, the mixture was centrifuged at 7012 g for 5 minutes to remove the supernatant and washed three times with 3.0 mL of fresh purified water. The mixture was lyophilized to obtain microparticles loaded with ropivacaine.

[0213] The results of particle size analysis of fine particles combined with ropivacaine using the above active fine particles are as shown in Fig. 5. According to the results of particle size analysis, D10 / D50 / D90 were 10.8 ㎛, 17.7 ㎛, and 25.5 ㎛, respectively, and the span value was 0.8, confirming that uniform particles were formed.

[0214] In addition, the SEM image measurement results are as shown in Fig. 6. In addition, the specific surface area determined by N- adsorption / desorption isotherm at -196°C using Micromeritics (TriStar II 3020 Version 3.02 / Micromeritics instrument corporation) was 6.2318 m 2 / g, and the pore volume (Pore Volume, P / P0=0.9921) is 0.0113 cm 3 / g, and the pore size (adsorption average pore diameter) is 3.6543 nm.

[0215] Additionally, the encapsulation rate for semaglutide was confirmed to be 59.7%.

[0216]

[0217] Preparation of sustained-release microparticles containing semaglutide 2

[0218] (1) Manufacturing of active microparticles

[0219] BSA and ABC, which are water-soluble channel forming agents, were mixed at a weight ratio of 1:1 (30 mg each) and dissolved in 1 mL of distilled water to prepare a first aqueous solution. 3 g (in the range) of PLGA 7525 polymer was dissolved in an organic solvent (Dimethyl chloride) to prepare an oily solution. The first aqueous solution and the oily solution were mixed using a homogenizer (Polytron, 12,000 rpm, 1 min) to prepare a dispersion solution.

[0220] A second aqueous solution was prepared by dissolving 0.5% PVA (Polyvinyl alcohol) in purified water. The prepared dispersion solution was injected into a membrane with a pore size of 10 μm using a pump at an injection rate of 1.0 mL / min, and the second aqueous solution was injected into a membrane with a pore size of 30.0 mL / min, respectively, to form an emulsion. The emulsion was solidified at 25°C for 24 hours, and the organic solvent was removed and freeze-dried.

[0221] (2) Semaglutide loading method

[0222] Microparticles (270 mg) and the activator, protamine sulfate (270 mg), were suspended in 0.1 mM HEPES buffer, pH 6.4 (1 mL) and stirred with a rotary mixer at 37°C for 24 hours to prepare a suspension solution containing active microparticles. Semaglutide (30 mg) was added to the suspension solution (pH 7.4 (0.75 mL)), suspended, and stirred with a rotary mixer at 37°C for 24 hours. An additional incubation was performed at 40°C to 42°C for 2 hours to control the initial release of the drug. Upon completion of the incubation, the supernatant was removed by centrifugation at 7012 g for 5 minutes, and washed three times with 3.0 mL of purified water. Microparticles containing semaglutide were obtained by lyophilization. The encapsulation rate of the microparticles was confirmed to be 90.2%.

[0223]

[0224] In-vitro drug release analysis of active microparticles

[0225] Active microparticles containing semaglutide were prepared by the method of manufacturing sustained-release microparticles containing semaglutide 2. The release behavior of the drug was confirmed using the Transwell method with 25 mg of the microparticles in 10 mM phosphate buffer (pH 7.4). The concentration of the eluted drug was measured using liquid chromatography (HPLC, Agilent, 1260 Infinity system). The mobile phase was 0.025 M disodium hydrogen phosphate buffer solution, acetonitrile gradient mode, and UV detection wavelength: 215 nm.

[0226] The test results are as shown in Fig. 7. According to Fig. 7, it was confirmed that the release effect of semaglutide was stable for up to 28 days.

[0227] Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

[0228] The present invention relates to a method for producing active microparticles according to the characteristics of a drug, active microparticles produced by the method for producing the same, and a method for producing a long-acting formulation comprising a drug using the active microparticles.

Claims

1. A step of preparing an oily solution by dissolving a biodegradable polymer in an organic solvent; A step of preparing an aqueous solution by dissolving a surfactant in water; A step of preparing an emulsion using the above oil solution and water solution; and A step of preparing active microparticles by freeze-drying the above emulsion. Method for producing active microparticles according to the properties of drugs.

2. In paragraph 1, The above-mentioned oil solution is a method for producing active microparticles according to the characteristics of a drug, which additionally includes an activator.

3. In paragraph 2, A method for producing active microparticles according to the characteristics of a drug, wherein the above activator is dissolved in an organic solvent together with a biodegradable polymer to produce an oily solution.

4. In paragraph 2, A method for producing active microparticles according to the characteristics of a drug, wherein the above-mentioned activator is dissolved in distilled water to produce a solution containing the activator, and a biodegradable polymer is mixed with an oily solution dissolved in an organic solvent to form a first emulsion.

5. In paragraph 2, The above activator is a method for producing active microparticles according to the properties of a drug that is a channel forming agent and / or a binding activator.

6. In paragraph 5, A method for producing active microparticles according to the characteristics of a drug, wherein the channel forming agent is selected from the group consisting of polyvinyl alcohol (PVA), ammonium bicarbonate (ABC), bovine serum albumin (BSA), trehalose, ethanol, pluronic, calcium chloride (CaCl2), surfactants, and mixtures thereof.

7. In paragraph 1, A method for producing active microparticles according to the characteristics of a drug, wherein the above binding activator is selected from the group consisting of polyethyleneimine (PEI), protamine sulfate, bovine serum albumin (BSA), calcium chloride (CaCl2), and mixtures thereof.

8. Active microparticles manufactured by the manufacturing method according to clauses 1 to 7.

9. A step of adding the active microparticles according to Article 8 to a buffer solution and stirring to prepare a suspension solution containing the active microparticles; and A method for producing a long-acting formulation comprising a drug using active microparticles, comprising the step of adding a drug to a suspension solution containing the active microparticles and stirring the drug to bind the drug to the active microparticles.

10. In paragraph 9, After the step of binding the drug to the above active microparticles, A method for producing a long-acting formulation containing a drug using active microparticles, the method further comprising performing an incubation step for 1 to 5 hours at a temperature higher than the Tg temperature of a biodegradable polymer included in the active microparticles.

11. In paragraph 9, The condition above the above Tg temperature is 40℃ or higher. A method for preparing a long-acting dosage form comprising a drug using active microparticles.

12. In paragraph 9, A method for producing a long-acting formulation containing a drug using active microparticles, wherein pores formed on the outside of the active microparticles are closed by self-healing through the above incubating step.

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