Sustained-release injection containing deslorelin and method for producing the same

Abstract

The present invention relates to a sustained-release injectable formulation comprising biodegradable polymeric microspheres containing deslorelin as an active ingredient, and a method for producing the same. The sustained-release microsphere injectable formulation containing deslorelin according to the present invention has good administration properties, and with a single administration, the effective concentration of deslorelin drug can be maintained in the blood of an animal for six months without sudden transient release.

Description

Technical Field

[0001] [Cross - reference to Related Applications] This application claims the benefit of priority based on Korean Patent Application No. 10 - 2018 - 0163416, filed on December 17, 2018, and all contents disclosed in the document of the Korean Patent Application are incorporated herein by reference.

[0002] The present invention relates to a biodegradable microsphere injection containing deslorelin and a method for producing the same.

[0003] The present invention relates to a sustained - release preparation and a veterinary preparation of deslorelin, which is an agent that acts on the peptide gonadotropin - releasing hormone. The use of the preparation includes, in particular, the prevention of reproductive functions in dogs and cats and the treatment of other diseases or conditions where suppression of prostate and breast cancer and testosterone or estradiol levels is beneficial.

Background Art

[0004] Promiscuous breeding of pet animals is a global problem, and it is particularly difficult to control the breeding of dogs and cats in developing countries. Surgical castration aims to regulate the reproduction of these animals. However, surgical operations carry risks, no matter how small. Many pet owners do not prefer to surgically deform their animals.

[0005] Therefore, peptide gonadotropin - releasing hormone has often been used for the purpose of suppressing reproductive functions and other processes that are affected by sex hormone values.

[0006] For a non-surgical neutering agent for companion animals to succeed industrially, it must be effective in all animals and the period of sex hormone suppression must be predictable. Such a period of sex hormone suppression must last for six months. That is, an efficient drug delivery system in which the peptide drug is released at a rate and volume that can predictably and stably block sex hormone secretion in animals for six months, and at a rate such that the proportion of the drug regulated for more than six months is released, must be provided. The reversibility of the inhibitory effect is an additional favorable advantage compared to surgical procedures.

[0007] Peptech in Australia has developed a lipid implant containing deslorelin (trade name: Suprelorin; distributor: Virbac in France) and sells it in 6-month and 12-month dosage forms for chemical castration of male dogs. However, Suprelorin has the disadvantage that a thick injection needle must be used when injecting animals in implant dosage form (6-month dosage form: 12 gauge, 12-month dosage form: 11 gauge). Also, it is known that the effect of the drug appears late initially and the drug duration is not constant. For these reasons, it has not been approved for neutering female dogs and cats.

Summary of the Invention

Problems to be Solved by the Invention

[0008] Therefore, there is a need to develop a sustained-release injection preparation of deslorelin that has a sufficient initial drug release amount when administered to companion animals such as dogs and cats, and in which the drug is released stably and constantly for a certain period, for example, for more than six months, and the drug release is completed after a certain period.

[0009] The inventors have developed a sustained-release preparation containing deslorelin as an active agent that exhibits an early onset of the effect of the initial drug when administered to animals and suppresses reproductive function over a predictable period. The dosage form is also a reversible neutralizing agent that restores reproductive function after the end of the drug release period following administration. The deslorelin preparation according to the present invention is useful for the treatment of a range of hormone-dependent diseases and conditions in animals, particularly companion animals such as dogs and cats. The preparation according to the present invention reduces the need for frequent subcutaneous injections or implant insertions by administering deslorelin once every six months, and provides an improved treatment method for hormone-dependent diseases and conditions in animals.

[0010] The present invention was devised to solve the problems of conventional surgical neutering and deslorelin implant preparations as described above, and aims to provide deslorelin sustained-release microspheres that have a sufficient initial release amount of the deslorelin drug, exhibit stable drug release characteristics for six months or more, and release 85% or more of the drug administered over six months, and a method for producing the same.

Means for Solving the Problems

[0011] To achieve the above object, the present invention uses two or more polymers selected from the group consisting of poly(lactide-co-glycolide) and polylactide polymers having a ratio of lactide to glycolide of 50:50 to 100:0 and an intrinsic viscosity of 0.16 to 1.2 dL / g, and provides a deslorelin sustained-release microsphere injection in which the deslorelin content is 5 to 25% by weight based on the total weight of the microspheres and a method for producing the same.

[0012] The present invention also provides a deslorelin sustained-release microsphere injection in which the average particle size of the deslorelin sustained-release microspheres is 10 μm to 100 μm, having a uniform particle size and good administrability, and a method for producing the same.

[0013] Hereinafter, the present invention will be described in detail.

[0014] The sustained-release microspheres of desloratadine of the present invention are manufactured using two or more polymers selected from the group consisting of poly(lactide-co-glycolide) and polylactide polymers, where the ratio of lactide to glycolide is from 50:50 to 100:0 and the intrinsic viscosity is 0.16 to 1.2 dL / g.

[0015] For the microspheres of desloratadine of the present invention, it is preferable to use two or more polymers selected from the group consisting of poly(lactide-co-glycolide) and polylactide polymers, where the ratio of lactide to glycolide is from 50:50 to 100:0, as the release regulator. The intrinsic viscosity of the poly(lactide-co-glycolide) or polylactide used in the present invention refers to the value measured at 25°C in chloroform at a concentration of 0.1% (w / v) using an Ubbelohde viscometer. When the intrinsic viscosity of the poly(lactide-co-glycolide) or polylactide is less than 0.16 dL / g, the molecular weight of the polymer is insufficient and it is difficult to exhibit the sustained-release effect of the desloratadine drug. When the intrinsic viscosity exceeds 1.2 dL / g, the effect that the release of the desloratadine drug is excessively delayed may appear. Also, when using a polymer with a high intrinsic viscosity, there is a problem that an excessive amount of the manufacturing solvent must be used due to the high viscosity of the polymer during microsphere production, and it is difficult to manufacture reproducible microspheres. Examples of commercially available polymers having the above-described characteristics include RG502H, RG503H, RG504H, RG502, RG503, RG504, RG653H, RG752H, RG752S, RG755S, RG750S, R202H, R203H, R205H, R202S, R203S, R205S, R206S, and R207S of the Resomer series of Evonik, and PDL 02A, PDL 02, PDL 04, PDL 05, PDLG 7502A, PDLG 7502, PDLG 7507, PDLG 5002A, PDLG 5002, PDLG 5004A, and PDLG 5004 of Corbion.

[0016] The content of deslorelin in the sustained-release microspheres containing deslorelin according to the present invention is preferably 5% to 25% by weight based on the total weight of the deslorelin microspheres. More preferably, it is 10% to 20% by weight. When the content of deslorelin in the microspheres is less than 5% by weight, the amount of polymer used is excessively large, resulting in a low bioavailability of deslorelin. When the content is higher than 25% by weight, there is a problem that the initial release of deslorelin increases, which is not preferable.

[0017] The microspheres containing deslorelin according to the present invention preferably have a uniform particle size distribution with an average particle size of 10 μm to 100 μm. The term "average particle size" used in the present invention means the median diameter, which is the particle size corresponding to 50% of the volume% in the particle size distribution curve, and is denoted as D50 or D(v,0.5).

[0018] When the average particle size of the microspheres containing deslorelin is less than 10 μm, the release of the deslorelin drug from the microspheres is excessively fast, which is not preferable. When the average particle size exceeds 100 μm, the injection needle becomes excessively thick during administration to animals, and there may be pain during injection and leakage of the drug at the injection site after injection, which is not preferable.

[0019] The microspheres containing desloratadine of the present invention preferably have a uniform particle distribution. Microspheres containing desloratadine with a uniform particle distribution have a smaller deviation during injection and can be administered in a more accurate amount compared to non-uniform microspheres. The size distribution or Span value of the microspheres containing desloratadine of the present invention is preferably 1.2 or less. More preferably, the size distribution is preferably 1.0 or less. The terms "size distribution" or "Span value" used in the present invention, as an index representing the uniformity of the particle size of the microspheres, mean the value obtained by the formula size distribution (Span value) = (Dv0.9 - Dv0.1) / Dv0.5. Here, Dv0.1 is the particle size corresponding to 10% of the volume% in the particle size distribution curve of the microspheres, Dv0.5 is the particle size corresponding to 50% of the volume% in the particle size distribution curve of the microspheres, and Dv0.9 is the particle size corresponding to 90% of the volume% in the particle size distribution curve of the microspheres.

[0020] When the sustained-release microspheres containing desloratadine of the present invention are administered to an animal by subcutaneous or intramuscular injection and no further release is desired after a certain period has elapsed, there is no way to recover them other than by surgical operation. Therefore, it is preferable that most of the drug is released within 6 months with the sustained-release microspheres containing desloratadine of the present invention. Accordingly, the sustained-release microspheres containing desloratadine of the present invention, although such a release pattern is not particularly limited, preferably release 85% or more of the drug within 6 months after being administered in vivo.

[0021] In a specific embodiment, the sustained-release microspheres containing desloratadine according to the present invention preferably release 10% or more of the drug within 1 month after administration and 85% or more within 6 months, more preferably 15% or more within 1 month after administration, 40% - 80% within 3 months, and 85% or more within 6 months, and most preferably 15% - 70% of the drug is released within 1 month after administration of the microspheres, 40% - 80% of the drug is released within 3 months, and 90% or more of the drug is released within 6 months.

[0022] Hereinafter, the manufacturing method of the sustained-release microsphere injection containing deslorelin of the present invention will be specifically described.

[0023] The sustained-release microsphere injection containing deslorelin according to the present invention is manufactured, for example, using the "solvent extraction and evaporation method", but the manufacturing method is not limited thereto.

[0024] As a specific example of the manufacturing method of the sustained-release microsphere containing deslorelin according to the present invention, such a manufacturing method comprises: (a) dissolving deslorelin and two or more polymers selected from the group consisting of poly(lactide-co-glycolide) and polylactide polymers in a mixed organic solvent to obtain As the dispersed phase deslorelin-polymer solution the liquid a manufacturing step; (b) adding the deslorelin-polymer solution produced in the step (a) to As the continuous phase an aqueous solution containing a surfactant in the phase to produce a dispersed phase in an emulsion state; (c) extracting and evaporating the organic solvent from the dispersed phase of the emulsion produced in the step (b) into the continuous phase to form microspheres; and (d) recovering the microspheres from the continuous phase of the step (c) to produce deslorelin microspheres. among them The manufacturing method includes the steps of: (a) dissolving deslorelin and two or more polymers selected from the group consisting of poly(lactide-co-glycolide) and polylactide polymers in a mixed organic solvent to obtain a deslorelin-polymer solution; (b) adding the deslorelin-polymer solution to an aqueous solution containing a surfactant to produce a dispersed phase in an emulsion state; (c) extracting and evaporating the organic solvent from the dispersed phase of the emulsion into the continuous phase to form microspheres; and (d) recovering the microspheres from the continuous phase to produce deslorelin microspheres.

[0025] For the sustained-release microspheres containing deslorelin according to the present invention, it is preferable to use two or more poly(lactide-co-glycolide) or polylactide polymers in order to achieve sufficient initial drug release for the early drug effect and drug release at a constant concentration for 6 months.

[0026] For the sustained-release microspheres containing deslorelin according to the present invention, by simultaneously dissolving deslorelin and two or more polymers selected from the group consisting of poly(lactide-co-glycolide) and polylactide polymers in a mixed organic solvent in the step (a), deslorelin sustained-release microspheres containing two or more polymers can be produced.

[0027] In addition, the sustained-release microspheres containing deslorelin according to the present invention can be produced by mixing two or more different microspheres containing deslorelin, which are produced through the above steps (a) to (d), using two or more different poly(lactide-co-glycolide) or polylactide polymers.

[0028] As a specific embodiment, a method for producing two or more different microspheres containing deslorelin by selecting two or more different polymers from the group consisting of poly(lactide-co-glycolide) and polylactide and using these may include the following steps.

[0029] Producing two or more different preliminary microspheres using two or more different polymers selected from the group consisting of poly(lactide-co-glycolide) or polylactide polymers and deslorelin; and mixing the two or more different preliminary microspheres.

[0030] At this time, the step of producing the two or more different preliminary microspheres may include the following steps: (a) dissolving deslorelin and poly(lactide-co-glycolide) or polylactide polymers in a mixed organic solvent to produce a deslorelin-polymer solution; (b) adding an aqueous solution containing a surfactant to the deslorelin-polymer solution produced in step (a) to produce a dispersed phase in an emulsion state; (c) extracting and evaporating the organic solvent from the dispersed phase of the emulsion produced in step (b) to form microspheres; and (d) recovering the microspheres from the continuous phase of step (c) to produce preliminary microspheres including the step of producing microspheres. As the dispersed phase deslorelin-polymer solution the liquid produced; (b) adding the deslorelin-polymer solution produced in step (a) to an aqueous solution containing a surfactant to produce a dispersed phase in an emulsion state; (c) extracting and evaporating the organic solvent from the dispersed phase of the emulsion produced in step (b) to form microspheres; and (d) recovering the microspheres from the continuous phase of step (c) to produce preliminary microspheres including the step of producing microspheres. As the continuous phase aqueous solution containing a surfactant in the phase to produce a dispersed phase in an emulsion state; (c) extracting and evaporating the organic solvent from the dispersed phase of the emulsion produced in step (b) to form microspheres; and (d) recovering the microspheres from the continuous phase of step (c) to produce preliminary microspheres including the step of producing microspheres. among them to form microspheres; and (d) recovering the microspheres from the continuous phase of step (c) to produce preliminary microspheres including the step of producing microspheres.

[0031] In a specific embodiment, the step of producing the two or more different preliminary microspheres may be performed sequentially or simultaneously.

[0032] In the step (a), the intrinsic viscosity of poly (lactide-co-glycolide) or polylactide is preferably in the range of 0.16 to 1.2 dL / g.

[0033] In the step (a), for the mixed solvent used to dissolve two or more polymers selected from the group consisting of deslorelin, poly (lactide-co-glycolide), and polylactide, it is preferable to use at least 50% (v / v) or more of a solvent having a property of being immiscible with water. By utilizing the property of the organic solvent being immiscible with water, in the subsequent step (b), the dispersed phase can be homogeneously mixed with the continuous phase containing a surfactant to form an emulsion. The type of such a mixed solvent for dissolving deslorelin and poly (lactide-co-glycolide) or polylactide is not particularly limited, but preferably a mixed solvent of two or more solvents selected from the group consisting of dichloromethane, chloroform, ethyl acetate, methyl ethyl ketone, acetone, acetonitrile, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, acetic acid, methyl alcohol, ethyl alcohol, propyl alcohol, and benzyl alcohol, and more preferably a mixed solvent of one solvent selected from dichloromethane and ethyl acetate and one or more solvents selected from dimethyl sulfoxide, N-methylpyrrolidone, and methyl alcohol can be used.

[0034] The method for homogeneously mixing the deslorelin-polymer solution and the continuous phase containing a surfactant in the step (b) is not particularly limited, but can be carried out by using a high-speed stirrer, an in-line mixer, a membrane emulsion method, a microfluidics emulsion method, etc. When forming an emulsion by using a high-speed stirrer or an in-line mixer, it is difficult to obtain a uniform emulsion, so it is preferable to additionally perform a sieving process or the like between the subsequent step (c) and step (d). When using the membrane emulsion method and the microfluidics emulsion method, an emulsion with a uniform size can be obtained, and it is more preferable because an additional sieving process or the like is not required between the subsequent step (c) and step (d).

[0035] The type of surfactant used in the step (b) is not particularly limited, and any surfactant can be used as long as it helps the deslorelin - polymer solution to form a stable dispersed phase of droplets in the continuous phase. The surfactant is preferably selected from the group consisting of methyl cellulose, polyvinyl pyrrolidone, carboxymethyl cellulose, lecithin, gelatin, polyvinyl alcohol, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene castor oil derivatives, and mixtures thereof, and most preferably polyvinyl alcohol can be used.

[0036] In the step (b), the content of the surfactant in the continuous phase containing the surfactant can be 0.01 w / v% to 20 w / v%, preferably 0.1 w / v% to 5 w / v% based on the total volume of the continuous phase containing the surfactant. When the content of the surfactant is less than 0.01 w / v%, a dispersed phase or emulsion in droplet form is not formed in the continuous phase. When the content of the surfactant exceeds 20 w / v%, it is difficult to remove the surfactant after fine particles are formed in the continuous phase by the excessive surfactant.

[0037] The continuous phase used in the step (b) can be water, and water containing one or more selected from the group consisting of methyl alcohol, ethyl alcohol, propyl alcohol, and ethyl acetate can be used to adjust the extraction rate of the organic solvent from the dispersed phase in the emulsion state.

[0038] In the step (c), when an emulsion containing a dispersed phase in droplet form and a continuous phase containing a surfactant is maintained or stirred at a temperature below the boiling point of the organic solvent for a certain period of time, for example, between 2 hours and 48 hours, the organic solvent can be extracted from the deslorelin - polymer solution in droplet form as the dispersed phase into the continuous phase. A part of the organic solvent extracted into the continuous phase can be evaporated from the surface. By extracting and evaporating the organic solvent from the deslorelin - polymer solution in droplet form, the dispersed phase in droplet form can be solidified to form microspheres.

[0039] In order to additionally and efficiently remove the organic solvent in step (c), heat can be applied to the continuous phase for a certain period of time.

[0040] In step (d), the method for recovering the microspheres containing deslorelin can be carried out using various known techniques, for example, methods such as filtration or centrifugation can be used.

[0041] Between step (c) and step (d), the remaining surfactant can be removed by filtration and washing, and then filtered again to recover the microspheres.

[0042] The washing step for removing the remaining surfactant can be carried out using normal water, and the washing step can be repeated several times.

[0043] Also, as described above, when an emulsion is formed using a high-speed stirrer and an in-line mixer in step (b), uniform microspheres can be obtained by additionally using a sieving process between step (c) and step (d). The sieving process can be carried out using known techniques, and sieves with different sizes can be used to separate microspheres of small and large particles to obtain microspheres of a uniform size.

[0044] In the production method of the present invention, after step (d) or after the filtration and washing steps, the obtained microspheres can be dried using a normal drying method to finally obtain dried microspheres.

[0045] By the production method of the present invention, a sustained-release microsphere injection containing deslorelin with uniform particles having good dosing ability and capable of maintaining an effective concentration in the animal's blood for 6 months without a rapid temporary release upon a single administration can be produced.

Effects of the Invention

[0046] The deslorelin sustained-release microsphere injection according to the present invention has good dosing ability and can maintain the effective concentration of deslorelin in the animal's blood for 6 months or more with a single administration.

Brief Description of Drawings

[0047]

Figure 1

Modes for Carrying Out the Invention

[0048] Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited by the following examples.

[0049] [Comparative Example] (Comparative Example 1.) Preparation 1 of a sustained-release microsphere injection preparation containing deslorelin produced using a single polymer The dispersed phase was prepared by mixing 9 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, and 1 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 32.14 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.59 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1 wt% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, washed repeatedly several times with distilled water to remove the residual polyvinyl alcohol, and freeze-dried to obtain microspheres containing deslorelin.

[0050] (Comparative Example 1-1.) Preparation 2 of a sustained-release microsphere injection preparation containing deslorelin produced using a single polymer The dispersed phase was prepared by mixing 8.5 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0051] (Comparative Example 1-2.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 3 The dispersed phase was prepared by mixing 2.4 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, and 0.6 of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 8.57 g of dichloromethane (manufacturer: J.T Baker, USA) and 2.56 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the dispersed phase at the same time to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by freeze-drying to obtain microspheres containing deslorelin.

[0052] (Comparative Examples 1 - 3.) Preparation of a sustained-release microsphere injection preparation containing deslorelin prepared using a single polymer 4 The dispersed phase was prepared by mixing 7.0 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, and 3.0 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 25.0 g of dichloromethane (manufacturer: J.T Baker, USA) and 7.46 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for over 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while simultaneously injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by freeze-drying to obtain microspheres containing deslorelin.

[0053] (Comparative Example 2.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 5 The dispersed phase was prepared by mixing 8.5 g of Resomer R202H (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0054] (Comparative Example 3.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 6 The dispersed phase was prepared by mixing 8.5 g of Resomer R205S (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0055] (Comparative Example 4.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 7 The dispersed phase was prepared by mixing 9.0 g of Resomer RG752H (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.0 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 32.14 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.59 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). It was connected to an emulsifying device equipped with the above-mentioned dispersed phase and a porous membrane (pore size: 20 μm), and while injecting, the prepared dispersed phase was injected to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0056] (Comparative Example 5.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 8 The dispersed phase was prepared by mixing 2.55 g of Resomer RG753H (manufacturer: Evonik, Germany), a biocompatible polymer, and 0.45 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 9.11 g of dichloromethane (manufacturer: J.T Baker, USA) and 2.72 ml of methyl alcohol (manufacturer: Tedia Company, USA). The dispersed phase was stirred for over 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 30 μm) and injected while simultaneously injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was complete, the temperature of the microsphere suspension was adjusted to 25°C and repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0057] (Comparative Example 6.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 9 The dispersed phase was prepared by mixing 8.5 g of Resomer RG653H (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while the dispersed phase was being injected to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0058] (Comparative Example 6-1.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 10 The dispersed phase was prepared by mixing 4.5 g of Resomer RG653H (manufacturer: Evonik, Germany), a biocompatible polymer, and 0.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 16.07 g of dichloromethane (manufacturer: J.T Baker, USA) and 4.8 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0059] (Comparative Example 7.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using a single polymer 11 The dispersed phase was prepared by mixing 8.5 g of Resomer RG503H (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0060] (Comparative Example 8.) Preparation of a sustained-release microsphere injection preparation containing deslorelin prepared using a single polymer 12 The dispersed phase was prepared by mixing 8.5 g of Resomer RG858S (manufacturer: Evonik, Germany), a biocompatible polymer, and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 60.71 g of dichloromethane (manufacturer: J.T Baker, USA) and 18.12 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for over 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by freeze-drying to obtain microspheres containing deslorelin.

[0061] (Comparative Example 9.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two kinds of polymers 1 Two kinds of polymers were used. In order to produce a sustained-release microsphere injection preparation having an average particle size of 100 μm or more using a high-speed stirrer, a deslorelin-containing sustained-release microsphere injection preparation was produced by the following method. The dispersed phase was prepared by mixing 1.91 g of Resomer R203H (manufacturer: Evonik, Germany), 0.64 g of Resomer RG752H (manufacturer: Evonik, Germany), and 0.45 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 9.11 g of dichloromethane (manufacturer: J.T Baker, USA) and 2.72 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for 30 minutes or more until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was added to the continuous phase and stirred at 1000 RPM using a high-speed stirrer to form an emulsion, which was stirred at 40 °C for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0062] (Comparative Example 10.) Preparation of a deslorelin-containing sustained-release microsphere injection preparation produced using two kinds of polymers 2 To produce a sustained-release microsphere injection preparation with an average particle size within 100 μm using two kinds of polymers, a deslorelin-containing sustained-release microsphere injection preparation was produced by the following method. To produce the sustained-release microsphere injection preparation with an average particle size within 100 μm, the RPM of the high-speed stirrer was relatively increased and produced as follows. The dispersed phase was prepared by mixing 1.91 g of Resomer R203H (manufacturer: Evonik, Germany), 0.64 g of Resomer RG752H (manufacturer: Evonik, Germany), and 0.45 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 9.11 g of dichloromethane (manufacturer: J.T Baker, USA) and 2.72 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved and then used. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was added to the continuous phase and stirred at 2000 rpm using a high-speed stirrer to form an emulsion, which was stirred at 40°C for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, washed repeatedly several times with distilled water to remove the residual polyvinyl alcohol, and freeze-dried to obtain microspheres containing deslorelin.

[0063] (Comparative Example 11.) Preparation of a deslorelin-containing sustained-release microsphere injection preparation produced using two kinds of polymers 3 Two polymers are used. The drug input into the production of the formulation is used at 40% by weight based on the total solid content. To produce a deslorelin-containing sustained-release microsphere injection formulation with a high final deslorelin content of 27.3% by weight in the sustained-release microsphere injection formulation, the sustained-release microsphere injection formulation was produced by the following method. The dispersed phase was prepared by mixing 1.35 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, 0.45 g of Resomer RG752H (manufacturer: Evonik, Germany), and 1.2 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 6.43 g of dichloromethane (manufacturer: J.T Baker, USA) and 1.92 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved and then used. The continuous phase used was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was injected while being connected to an emulsifying device equipped with a porous membrane (pore size: 50 μm) to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0064] (Comparative Example 12.) Preparation of a deslorelin-containing sustained-release microsphere injection formulation produced using two polymers 4 Two kinds of polymers are used, and the drug input into the production of the preparation is used at 5% by weight based on the total solid content. To produce a sustained-release microsphere injection preparation containing deslorelin with a low final content of 4.2% by weight, the sustained-release microsphere injection preparation was produced by the following method. The dispersed phase was prepared by mixing 2.14 g of Resomer R203H (manufacturer: Evonik, Germany), 0.71 g of Resomer RG752H (manufacturer: Evonik, Germany), and 0.15 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 10.18 g of dichloromethane (manufacturer: J.T Baker, USA) and 3.04 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved and then used. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0065] (Comparative Example 13.) Preparation of a deslorelin-containing sustained-release microsphere injection preparation produced using two kinds of polymers 5 Two kinds of polymers were used. In order to produce a sustained-release microsphere injection preparation containing desloratadine with the ratio of PLA to PLGA (PLA:PLGA) being 1:3, the sustained-release microsphere injection preparation was produced by the following method. The dispersed phase was prepared by mixing 1.06 g of Resomer R203H (manufacturer: Evonik, Germany), 3.19 g of Resomer RG752H (manufacturer: Evonik, Germany), and 0.75 g of desloratadine acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 15.18 g of dichloromethane (manufacturer: J.T Baker, USA) and 4.53 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing desloratadine.

Example

[0066] In the example, when producing a sustained-release microsphere injection preparation containing desloratadine, two or more kinds of polymers were used, or two or more dosage forms produced using a single polymer were mixed.

[0067] (Example 1.) Preparation 1 of a sustained-release microsphere injection preparation containing desloratadine produced using two or more kinds of polymers The dispersed phase was prepared by mixing 6.375 g of Resomer R203H (manufacturer: Evonik, Germany), 2.125 g of Resormer RG858S (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 37.95 g of dichloromethane (manufacturer: J.T Baker, USA) and 11.33 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while the above-mentioned dispersed phase was injected to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0068] (Example 2.) Preparation of a sustained-release microsphere injection preparation containing deslorelin prepared using two or more polymers 2 The dispersed phase was prepared by mixing 7.65 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, 0.85 g of Resormer RG752H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was injected while being connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0069] (Example 3.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 3 The dispersed phase was prepared by mixing 6.375 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG753H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0070] (Example 4.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 4 The dispersed phase was prepared by mixing 4.25 g of Resomer R203H (manufactured by Evonik, Germany), 4.25 g of Resormer RG752H (manufactured by Evonik, Germany), and 1.5 g of deslorelin acetate (manufactured by Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufactured by J.T Baker, USA) and 9.06 ml of methyl alcohol (manufactured by TEDIA Company, USA). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected. At the same time, the prepared dispersed phase was injected to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0071] (Example 5.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 5 The dispersed phase was prepared by mixing 6.375 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG653H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0072] (Example 6.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 6 The dispersed phase was prepared by mixing 6.375 g of Resomer R203H (manufacturer: Evonik, Germany), 2.125 g of Resormer RG503H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 14.73 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 50 μm) and injected, and at the same time, the prepared dispersed phase was injected to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0073] (Example 7.) Preparation of a sustained-release microsphere injection preparation containing deslorelin prepared using two or more polymers 7 The dispersed phase was prepared by mixing 6.375 g of Resomer R202H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG858S (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 37.95 g of dichloromethane (manufacturer: J.T Baker, USA) and 11.33 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while simultaneously injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by freeze-drying to obtain microspheres containing deslorelin.

[0074] (Example 8.) Preparation of a sustained-release microsphere injection formulation containing deslorelin manufactured using two or more polymers 8 The dispersed phase was prepared by mixing 6.375 g of Resomer R202H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG752H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of methyl alcohol (manufacturer: TEDIA Company, USA). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected, and at the same time, the prepared dispersed phase was injected to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0075] (Example 9.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 9 The dispersed phase was prepared by mixing 6.375 g of Resomer R202H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG653H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 20 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0076] (Example 9-1.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 9-1 The dispersed phase was prepared by mixing 2.775 g of Resomer R202H (manufacturer: Evonik, Germany), a biocompatible polymer, 1.85 g of Resormer RG653H (manufacturer: Evonik, Germany), and 0.375 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 16.52 g of dichloromethane (manufacturer: J.T Baker, USA) and 4.93 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase used was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 5 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by lyophilization to obtain microspheres containing deslorelin.

[0077] (Example 9-2.) Preparation of a sustained-release microsphere injection preparation containing deslorelin prepared using two or more polymers 9-2 The dispersed phase was prepared by mixing 2.25 g of Resomer R202H (manufacturer: Evonik, Germany), 1.5 g of Resormer RG653H (manufacturer: Evonik, Germany), and 1.25 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 13.39 g of dichloromethane (manufacturer: J.T Baker, USA) and 4.00 ml of dimethyl sulfoxide (manufacturer: Samchun Pure Chemical, South Korea). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase used was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 5 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40 °C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by freeze-drying to obtain microspheres containing deslorelin.

[0078] (Example 10.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 10 The dispersed phase was prepared by mixing 6.375 g of Resomer R202H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG503H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The above-mentioned dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 40 μm) and injected while injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, and then freeze-dried to obtain microspheres containing deslorelin.

[0079] (Example 11.) Preparation of a sustained-release microsphere injection preparation containing deslorelin manufactured using two or more polymers 11 The dispersed phase was prepared by mixing 4.25 g of Resomer R203H (manufacturer: Evonik, Germany), a biocompatible polymer, 2.125 g of Resormer RG752H (manufacturer: Evonik, Germany), 2.125 g of Resormer RG503H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 30.36 g of dichloromethane (manufacturer: J.T Baker, USA) and 9.06 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). The prepared dispersed phase was connected to an emulsifying device equipped with a porous membrane (pore size: 40 μm) and injected while simultaneously injecting the prepared dispersed phase to form an emulsion. This emulsion was stirred at 40°C and 200 RPM for 3 hours to remove the organic solvent. After the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25°C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol, followed by freeze-drying to obtain microspheres containing deslorelin.

[0080] (Example 12 - 22.) Preparation of a formulation containing two or more types of microspheres containing different polymers The sustained-release microsphere formulation containing deslorelin prepared in the above comparative example was mixed by polymer type and ratio as shown in Table 1 below to obtain Examples 12 - 22.

[0081]

Table 1

[0082] (Example 23.) Preparation of a sustained-release microsphere injection formulation containing deslorelin prepared using two or more polymers 12 The dispersed phase was prepared by mixing 4.25 g of Resomer R207S (manufacturer: Evonik, Germany), a biocompatible polymer, 4.25 g of Resormer RG752H (manufacturer: Evonik, Germany), and 1.5 g of deslorelin acetate (manufacturer: Chengdu Kaijie Biopharm Co., Ltd, China) with 42.5 g of dichloromethane (manufacturer: J.T Baker, USA) and 12.69 ml of N-methylpyrrolidone (manufacturer: JUNSEI, Japan). The dispersed phase was stirred for more than 30 minutes until completely dissolved before use. The continuous phase was an aqueous solution of 1% polyvinyl alcohol (viscosity: 4.8 - 5.8 mPa·s). After adding the above-mentioned dispersed phase to the continuous phase, it was stirred at 2000 RPM using a high-speed stirrer to form an emulsion, which was then stirred at 40 °C for 3 hours to remove the organic solvent. When the removal of the organic solvent was completed, the temperature of the microsphere suspension was adjusted to 25 °C, and it was repeatedly washed several times with distilled water to remove the residual polyvinyl alcohol. After removing the particles larger than 150 μm and smaller than 25 μm using 100-mesh (150 μm) and 500-mesh (25 μm) sieves, it was freeze-dried to obtain microspheres containing deslorelin.

[0083] [Experimental Example] (Experimental Example 1.) Measurement of deslorelin content in microspheres To measure the encapsulation amount of deslorelin in the microspheres prepared in Comparative Examples 1 - 13 and Examples 1 - 11, 2 mg of the microspheres were completely dissolved in acetonitrile and then diluted with the mobile phase. 20 μL of the diluted solution was injected into HPLC and measured at a detection wavelength of 280 nm. The column used for this measurement was Gemini C18 5 μm, 4.6 x 150 mm, and the mobile phase was (A) 0.1% aqueous TFA solution and (B) 0.1% TFA acetonitrile used in Gradient mode with (B) 20 - 50%. The deslorelin content in the microspheres prepared in the comparative examples is as shown in Table 2 below.

[0084]

Table 2

[0085] In Comparative Examples 1-8, a dosage form was manufactured using a single polymer. At this time, the types of polymers and the target drug content were varied during manufacturing. Based on the results shown in Table 2, it was confirmed that there were differences in the drug encapsulation rate depending on the type of polymer. Generally, when the end group of the polymer used for manufacturing the dosage form was an ester group, it was confirmed that the drug encapsulation rate tended to be relatively low. Comparative Examples 9-10 are examples where a preparation was manufactured using a high-speed stirrer, and it was confirmed that even when the manufacturing method was different, there was no significant effect on the drug content. Also, Comparative Examples 11-12 are preparations manufactured with different amounts of deslorelin relative to the total solid content introduced during microsphere production. It was confirmed that the final drug content varied significantly depending on the drug input amount, and it was confirmed that the higher the drug input ratio, the lower the encapsulation rate. Comparative Example 13 is an example where a preparation was manufactured using two types of polymers, with the mixing ratio being PLA:PLGA = 1:3, and it was confirmed that there was no difference in the drug encapsulation rate depending on the polymer composition ratio.

[0086] The content of deslorelin in the microspheres of Examples 1-23 is shown in Table 3 below.

[0087]

Table 3

[0088] (Experimental Example 2.) Morphological Analysis of Deslorelin-Containing Sustained-Release Microsphere Injection Preparation by Electron Microscope Scanning electron microscope observation was carried out to analyze the morphological characteristics of the microspheres. After the microspheres were platinum-coated using an ION-COATER (COXEM, Korea), the morphological characteristics of the microspheres were observed at an acceleration voltage of 15 kV using a scanning electron microscope (COXEM EM-30, Korea), and the results are shown in Figure 1.

[0089] Figure 1 is a photograph of the deslorelin-containing sustained-release microsphere injection preparation manufactured in Example 3 observed with an electron microscope. According to Figure 1, it was confirmed that microspheres with very uniform particle sizes were manufactured.

[0090] (Experimental Example 3.) Particle Size Analysis of Deslorelin-Containing Sustained-Release Microsphere Injection Formulation Using Laser Diffraction Method To quantitatively measure the average particle size, distribution, and uniformity of the microspheres, tests were conducted using the laser diffraction method. The microspheres produced in the comparative example and the examples were mixed with ultrapure water containing a surfactant using a vortex mixer for 20 seconds, and then placed in an ultrasonic generator for dispersion. This solution was placed in a particle size analyzer (Microtrac Bluewave, Japan) to measure the particle size.

[0091] The span value, as an index of particle size magnitude uniformity, was determined by the following formula 1.

[0092] [Formula 1] Span Value = (Dv0.9 - Dv0.1) / Dv0.5

[0093] The particle size measurement results of the deslorelin-containing sustained-release formulation produced in the comparative example are shown in Table 4 below.

[0094] [Table 4]

[0095] The particle size measurement results of the deslorelin-containing sustained-release microsphere formulation produced in the examples are shown in Table 5 below.

[0096] [Table 5]

[0097] (Experimental Example 4.) Injection Ability Test of Deslorelin-Containing Sustained-Release Microsphere Injection Formulation This experiment was conducted to examine the preferred average particle size of the microspheres by measuring the microsphere administration recovery rate. The specific experimental procedure is as follows.

[0098] 60 mg of the microspheres of the comparative examples and examples described in Table 6 below were each weighed, placed in a 1.5 mL vial, and mixed with 0.8 mL of distilled water. After collecting as much of the microsphere dispersion as possible using a 1 mL syringe equipped with a 25G injection needle, the 1.5 mL vial was dried and the weight of the non-recovered microspheres was measured. The recovery rate was calculated by dividing the weight of the microspheres excluding the measured weight of the non-recovered microspheres by the initial amount of microspheres used, 200 mg.

[0099]

Table 6

[0100] According to the above results, in Examples 18 and 19 and Comparative Examples 8, 10, and 13, where the particle size was 100 μm or less and the span value was 1.2 or less, high recovery rates of 84.2% (w / w) to 94.6% (w / w) were shown in the recovery rate experiment using a syringe equipped with a 25G injection needle. However, it was confirmed that even when having a low span value as in Comparative Example 9, if the average particle size exceeded 100 μm, the microspheres blocked the injection needle and could not be recovered smoothly. Also, even when the average particle size was 100 μm or less as in Comparative Example 6, when the span value was 1.2 or more, the particle size of the microspheres was not uniform, and thus the recovery rate of the microspheres was very low in the 50% (w / w) range.

[0101] As a result, it was confirmed that both the particle size and the span value affected the injectability, and particularly when the particle size was 100 μm or less and the span value was 1.2 or less, it had better injectability than microspheres that were not such.

[0102] (Experimental Example 5.) Single-dose subcutaneous administration pharmacokinetics study using Sprague-Dawley Rats To evaluate the pharmacokinetics of the deslorelin-containing sustained-release microsphere injection preparation, the blood deslorelin concentration was measured in rats over time after dosing. The deslorelin dosage volume was prepared by measuring microspheres to be 4.7 mg / head, dispersing them in 0.3 mL of suspension, and then subcutaneously injecting them into SD rats. 0.5 mL of blood was collected at pre-planned time intervals, and the deslorelin and testosterone concentrations in the blood were measured using LC-MS / MS.

[0103] The test results of the pharmacokinetics in rats of the dosage forms manufactured in the comparative examples and the examples were converted into the cumulative release rate with respect to AUC (area under the curve) and summarized in Table 7 below.

[0104]

Table 7

[0105] From the above results, in the case of the biodegradable polymer microsphere dosage form containing deslorelin manufactured with a single polymer, it was confirmed that the drug release in the dosage form manufactured using poly-lactide-co-glycolide was excessively short. For the dosage form manufactured using polylactide, although the period for maintaining the blood drug concentration relatively increased, it was confirmed that not only was there a period during which almost no drug was released after the initial release within the first month after administration, but also the bioavailability was significantly low.

[0106] However, in the case of the dosage form manufactured according to the examples of the present invention to overcome such problems, it was confirmed that the drug was continuously released for 6 months or more after administration. Although the release pattern of such a drug is not particularly limited, preferably 10% or more of the drug is released within 1 month after administration, 85% or more is released at 6 months, more preferably 15% or more is released within 1 month after administration, 40% to 80% is released at 3 months, 85% or more is released at 6 months, and most preferably 15% to 70% of the drug is released within 1 month after administration of the microspheres, 40% to 80% is released at 3 months, and 90% or more of the drug is released at 6 months.

Claims

1. Deslorelin at 5 to 25% by weight based on the weight of the total microparticles, and containing poly(lactide-co-glycolide) and polylactide polymers, wherein the average particle size of the microparticles is 10 to 100 μm and the span value is 1.2 or less, the intrinsic viscosity of the poly(lactide-co-glycolide) or polylactide is 0.16 to 1.2 dL / g, when the sustained-release microparticles containing deslorelin are administered subcutaneously or intramuscularly to an animal, 85% or more of the deslorelin in the microparticles is released over 6 months, the sustained-release microparticles containing deslorelin are characterized in that they are produced using a mixed solvent of (i) dichloromethane and (ii) one solvent selected from the group consisting of dimethyl sulfoxide, N-methylpyrrolidone, and methyl alcohol.

2. The sustained-release microparticles containing deslorelin according to claim 1, wherein the poly(lactide-co-glycolide) and polylactide polymers of the sustained-release microparticles containing deslorelin are contained in one kind of microparticles or independently contained in two or more kinds of microparticles respectively.

3. The sustained-release microparticles containing deslorelin according to claim 1 or 2, wherein the recovery rate at the time of injection of the microparticles is 80 to 100% when measured using a 1 mL syringe equipped with a 25G injection needle.

4. The sustained-release microparticles containing deslorelin according to claim 1, wherein 10% or more of the drug is released within 1 month after administration and 85% or more is released at 6 months.

5. (a) Dissolving deslorelin, poly(lactide-co-glycolide), and polylactide polymers in a mixed organic solvent to produce a deslorelin-polymer solution as a dispersion phase; (b) Adding the deslorelin-polymer solution produced in step (a) to an aqueous solution phase containing a surfactant as a continuous phase to produce an emulsion; (c) Extracting and evaporating the organic solvent from the dispersion phase of the emulsion produced in step (b) into the continuous phase to form microparticles; and (d) Recovering the microparticles from the continuous phase of step (c) to produce deslorelin microparticles, wherein the mixed organic solvent in step (a) is a mixed solvent of (i) dichloromethane and (ii) one solvent selected from the group consisting of dimethyl sulfoxide, N-methylpyrrolidone, and methyl alcohol. The sustained-release microspheres containing deslorelin have an average particle size of 10 to 100 μm and a span value of 1.2 or less, characterized in that the intrinsic viscosity of the poly(lactide-co-glycolide) or polylactide is 0.16 to 1.2 dL / g, A method for producing sustained-release microspheres containing deslorelin, wherein when the produced sustained-release microspheres containing deslorelin are administered by subcutaneous or intramuscular injection to an animal, 85% or more of the deslorelin in the microspheres is released over 6 months. **Claim 6**: A step of producing preliminary microspheres using poly(lactide-co-glycolide) and deslorelin, and a step of producing preliminary microspheres using a polylactide polymer and deslorelin; and a step of mixing two or more different preliminary microspheres, At this time, the step of producing the above-mentioned preliminary microspheres includes the following steps, a method for producing sustained-release microspheres containing deslorelin: (a) A step of dissolving deslorelin and a poly(lactide-co-glycolide) or polylactide polymer in a mixed organic solvent to produce a deslorelin-polymer solution as a dispersed phase; (b) A step of adding the deslorelin-polymer solution produced in the step (a) to an aqueous solution phase containing a surfactant as a continuous phase to produce a dispersed phase in an emulsion state; (c) A step of extracting and evaporating the organic solvent from the dispersed phase of the emulsion produced in the step (b) into the continuous phase to form microspheres; and (d) A step of recovering the microspheres from the continuous phase of the step (c) to produce microspheres, The mixed organic solvent in the step (a) is a mixed solvent of (i) dichloromethane and (ii) one solvent selected from the group consisting of dimethyl sulfoxide, N-methylpyrrolidone, and methyl alcohol, The sustained-release microspheres containing deslorelin have an average particle size of 10 to 100 μm and a span value of 1.2 or less, characterized in that the intrinsic viscosity of the poly(lactide-co-glycolide) or polylactide is 0.16 to 1.2 dL / g, A step of producing preliminary microspheres, wherein when the produced sustained-release microspheres containing deslorelin are administered by subcutaneous or intramuscular injection to an animal, 85% or more of the deslorelin in the microspheres is released over 6 months. **Claim 7** The production method according to claim 5 or 6, further including a sieving step between the step (c) and the step (d). **Claim 8** The surfactant in the step (b) is at least one selected from the group consisting of methyl cellulose, polyvinyl pyrrolidone, carboxymethyl cellulose, lecithin, gelatin, polyvinyl alcohol, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene castor oil derivative, and mixtures thereof. The manufacturing method according to claim 5 or 6.

9. The continuous phase in the step (b) is water or a mixed solvent of water and at least one selected from the group consisting of methyl alcohol, ethyl alcohol, propyl alcohol, and ethyl acetate. The manufacturing method according to claim 5 or 6.

10. The recovery rate during injection of the sustained-release microspheres containing deslorelin produced is 80 to 100% when measured using a 1 mL syringe equipped with a 25G injection needle. The manufacturing method according to claim 5 or 6.

11. The sustained-release microspheres containing deslorelin produced release 10% or more of the drug within 1 month and 85% or more within 6 months after administration. The manufacturing method according to claim 5 or 6.

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