Pharmaceutical composition comprising sustained-release microsphere comprising cagrilintide or pharmaceutically acceptable salt thereof and preparation method therefor

Abstract

The present invention relates to a pharmaceutical composition which, by comprising sustained-release microspheres which consist of cagrilintide or a pharmaceutically acceptable salt thereof, an initial-burst inhibitor, and a biodegradable polymer, does not undergo rapid initial burst of a drug, has a high content of the drug relative to the particle size, and has high bioavailability, thus being able to minimize the patient's pain and inflammatory response that may occur when administered to the human body, and thus is effective in the prevention or treatment of diabetes, beta-cell dysfunction, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, and Alzheimer's disease.

Description

Sustained-release microsphere preparation comprising caprylintide or a pharmaceutically acceptable salt thereof and method for preparing the same

[0001] The present invention relates to sustained-release microspheres comprising a high content of caprilintide or a pharmaceutically acceptable salt thereof, sustained-release microspheres simultaneously comprising caprilintide and semaglutide or a pharmaceutically acceptable salt thereof, or a blending of sustained-release microspheres comprising caprilintide or a pharmaceutically acceptable salt thereof and sustained-release microspheres comprising semaglutide or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the microspheres or the combination of microspheres and a method for preparing the sustained-release microspheres.

[0002] Amylin is a 37-amino acid peptide produced by pancreatic beta cells and secreted along with insulin. Amylin plays a crucial role in appetite, glucose management, and gastric emptying, warranting investigation in the treatment of diabetes and obesity. Human amylin is also involved in the formation of cytotoxic amyloids observed in the islets of Langerhans in patients with type 2 diabetes. In 2005, a modified amylin (pramlintide, discovered by Amylin Pharmaceuticals) was approved for use in combination with insulin for the treatment of both type 1 and type 2 diabetes. Pramlintide contains three proline substitutions compared to human amylin. Pramlintide has a short half-life (20–45 minutes) and requires frequent subcutaneous injections three times daily, which necessitates frequent dosing. Pramlintide has consistently induced weight loss in several trials, either alone or in combination with leptin, but has been associated with adverse effects such as nausea and vomiting at the start of treatment.

[0003] Meanwhile, Cagrilintide is a recently developed long-acting amylin analog. It has also been shown to be effective in combination with the GLP-1 agonist semaglutide to achieve sustained weight loss in overweight and obese individuals. Amylin, released along with insulin from pancreatic beta cells, induces satiety through the homeostatic and hedonic regions of the brain. Semaglutide, a GLP-1 receptor agonist, reduces appetite, increases insulin production, and decreases glucagon secretion via the hypothalamic GLP-1 receptor, delaying gastric emptying. These separate but related mechanisms of action of amylin analogs and GLP-1 receptor agonists appear to have an additive effect on appetite reduction. Given the heterogeneity and complex pathogenesis of obesity, combination therapy using multiple pathophysiological targets is a logical approach to augment the weight loss response to pharmacological therapy, and capriglintide, alone and in combination with semaglutide, has shown promising weight loss in clinical trials supporting further development of this regimen for sustained weight management.

[0004] However, for effective single or combined administration of kagrillintide, development of a technology to ensure long-term effective pharmacological effects is still necessary. Accordingly, the present inventors have devised a technology to encapsulate kagrillintide in microspheres made of biodegradable polymers for long-term dissolution. However, in this case, the bioavailability of kagrillintide encapsulated in the microspheres is low or the drug content contained within the microspheres is low, so large doses of microspheres must be administered to exhibit long-term effective pharmacological effects. However, when large doses of microspheres are administered in vivo, subcutaneous injection is difficult, making it difficult for patients to directly administer (self-administer) the drug, and there are also problems such as significantly increased pain and inflammatory reactions at the injection site.

[0005] The present invention has been proposed to solve the above problems, and its purpose is to provide sustained-release microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof, a biodegradable polymer, and an early-release inhibitor, which contain a high content of kagrillintide, and exhibit high bioavailability of kagrillintide and long-term stable drug release characteristics, sustained-release microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof, a biodegradable polymer, and an early-release inhibitor, or a combination of sustained-release microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof, a biodegradable polymer, and an early-release inhibitor, and sustained-release microspheres comprising semaglutide or a pharmaceutically acceptable salt thereof, a biodegradable polymer, and an early-release inhibitor, and a pharmaceutical composition comprising them, and a method for preparing the sustained-release microspheres.

[0006] The present invention is described in detail below.

[0007]

[0008] The term "one or more" in the present invention refers to a number corresponding to one or more. In the present invention, when a certain configuration is one or more, it may preferably be one, two or more, three or more, one to three, or one to two, but is not limited thereto. The term "one or more" may be used interchangeably with the term "one or more" in the present invention.

[0009]

[0010] To achieve the above purpose,

[0011] In one aspect, the present invention provides a pharmaceutical composition comprising kagrillintide sustained-release microspheres, which comprise kagrillintide or a pharmaceutically acceptable salt thereof as an active ingredient, a biodegradable polymer, and an early-release inhibitor, wherein the content of kagrillintide is 8 wt% or more as kagrillintide based on the total weight of the microspheres, and the early-release inhibitor is 10 ppm to 2000 ppm based on the total weight of the microspheres.

[0012] In one aspect, the present invention provides a pharmaceutical composition comprising kagrillintide sustained-release microspheres, which comprises kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof as active ingredients, a biodegradable polymer, and an early-release inhibitor, wherein the content of kagrillintide is 8 wt% or more as kagrillintide based on the total weight of the microspheres, the content of semaglutide is 8 wt% or more as semaglutide based on the total weight of the microspheres, and the early-release inhibitor is 10 ppm to 2000 ppm based on the total weight of the microspheres.

[0013] In one aspect, the present invention provides a pharmaceutical composition comprising a combination of caprilintide sustained-release microspheres comprising caprilintide or a pharmaceutically acceptable salt thereof as an active ingredient, a biodegradable polymer, and an early-release inhibitor, wherein the caprilintide content is 8 wt% or more as caprilintide based on the total weight of microspheres, the semaglutide content is 8 wt% or more as semaglutide based on the total weight of microspheres, and the early-release inhibitor is 10 ppm to 2000 ppm based on the total weight of microspheres, and semaglutide sustained-release microspheres comprising semaglutide or a pharmaceutically acceptable salt thereof as an active ingredient, a biodegradable polymer, and an early-release inhibitor.

[0014]

[0015] In another aspect, the biodegradable polymer is polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA), polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkylcarbonate, polytrimethylenecarbonate (PTMC), polylactide-co-trimethylenecarbonate (PLTMC), A polymer selected from the group consisting of block copolymers of polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo polyaminoacid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphoester polysaccharide and poly(butylene succinate lactide) (PBSLA); simple mixtures of two or more thereof; A copolymer of the above polymer and polyethylene glycol (polyethylenglycol, PEG);And it may be at least one selected from the group consisting of a polymer-sugar complex in which the polymer or copolymer is combined with a sugar.;

[0016] In another aspect, the pharmaceutically acceptable salt of the above-mentioned kagrillintide may be sodium salt, acetate salt, benzoate salt, hydroxynaphthoate salt, napadicylate salt or pamoate salt of kagrillintide.

[0017] In another aspect, the biodegradable polymer may be characterized by having an intrinsic viscosity of 0.16 to 1.7 dL / g.

[0018] In another aspect, the average particle size of the microspheres may be 5 μm to 100 μm.

[0019] In another aspect, the span value of the microsphere may be characterized as being 1.5 or less.

[0020] In another aspect, the weight of the microspheres is 20 to 1000 mg, 20 to 800 mg, 20 to 600 mg, 20 to 400 mg, 20 to 200 mg, 20 to 100 mg, 30 to 1000 mg, 30 to 800 mg, 30 to 600 mg, 30 to 400 mg, 30 to 200 mg, 30 to 100 mg, 40 to 1000 mg, 40 to 800 mg, 40 to 600 mg, 40 to 400 mg, 40 to 200 mg, 40 to 100 mg, 50 to 1000 mg, 50 to 800 mg, 50 to It may be 600 mg, 50 mg to 400 mg, 50 mg to 200 mg, or 50 mg to 100 mg.

[0021] In another aspect, it may further comprise one or more release controlling agents selected from butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, nonadecylic acid, behenic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, napadicylic acid, naphthalene sulfonic acid and pamoic acid.

[0022] In another aspect, the microspheres may comprise 10 to 500 mg / kg of Na.

[0023] In another aspect, the microspheres may comprise 1 to 100 mg / kg of P.

[0024] In another aspect, the present invention provides a method for preparing sustained-release microspheres containing (i) kagrillintide or a pharmaceutically acceptable salt thereof or (ii) kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof; a biodegradable polymer; and an early-release inhibitor as an active ingredient.

[0025] In another aspect, the manufacturing method may use a continuous phase including an early release inhibitor.

[0026] In another aspect, any substance that can be dissolved in the continuous phase during the manufacture of the microspheres and set the pH of the continuous phase to 7.0 or higher can be used as the initial release inhibitor.

[0027] In another aspect, the early-release inhibitor may use at least one substance selected from the group consisting of phosphate salts, hydroxide salts, phosphide salts, phosphite salts, carbonate salts, bicarbonate salts, chromate salts, dichromate salts, oxides, oxalate salts, silicate salts, sulfate salts, sulfide salts, sulfite salts, tartrate salts, tetraborate salts, thiosulfate salts, arsenate salts, arsenite salts, citrate salts, ferricyanide salts, and nitride salts of alkali metals, alkaline earth metals, or ammonium. In another aspect, the early-release inhibitor may be at least one substance selected from the group consisting of disodium phosphate, dipotassium phosphate, and diammonium phosphate.

[0028] In another aspect, a pharmaceutical composition comprising sustained-release microspheres comprising a biodegradable polymer and having a kagrillintide content of at least 8 wt% as kagrillintide based on the total weight of the microspheres and an initial drug release of at most 10%, and a method for preparing the same, are provided.

[0029] A sustained-release pharmaceutical composition comprising kagrillintide, a pharmaceutically acceptable salt thereof, and an early-release inhibitor according to one manufacturing example of the present invention has the effect of minimizing side effects due to early release when administered into the body by suppressing rapid early release despite containing a high drug content compared to particle size, while exhibiting high bioavailability and long-term sustained efficacy of more than one month, and allowing a reduced dosage, thereby minimizing pain in the patient and side effects at the injection site that may occur during administration.

[0030] Figure 1 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 1-1 to 1-7.

[0031] Figure 2 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 2-1 to 2-4.

[0032] Figure 3 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 3-1 to 3-4.

[0033] Figure 4 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 4-1 to 4-3.

[0034] Figure 5 is a scanning electron microscope photograph confirming the morphological characteristics of co-encapsulation microspheres manufactured in Examples 5-1 and 5-6.

[0035] Figure 6 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Comparative Examples 1 and 2.

[0036] The present invention is described in detail below.

[0037] The present invention comprises, as an active ingredient, kagrillintide or a pharmaceutically acceptable salt thereof.

[0038] Cagrilintide is a long-acting amylin analogue that targets the calcitonin receptor, a receptor activator of RAMP 1-3 (RAMP1-3). Cagrilintide is a non-selective agonist designed for once-weekly subcutaneous administration at low pH.

[0039] The structure of kagrillintide is as shown in chemical formula 1 below.

[0040] [Chemical Formula 1]

[0041]

[0042]

[0043] ​Kagrilintide may exist in the form of a salt, particularly a pharmaceutically acceptable salt. Any salt commonly used in the art may be used without limitation. The term "pharmaceutically acceptable salt" of the present invention refers to any and all organic or inorganic addition salts of the compound, which are relatively non-toxic and harmless to patients and have an effective effect, and whose side effects due to the salt do not diminish the beneficial effects of the active ingredient. Specific examples include, but are not limited to, the sodium salt, acetate salt, benzoate salt, hydroxynaphthoate salt, napadisylate salt, or pamoate salt of kagrillintide.

[0044] Kagrilintide, the active ingredient of the present invention, or a pharmaceutically acceptable salt thereof may be in various forms, for example, amorphous or crystalline.

[0045] The pharmaceutical composition according to the present invention has an area under the blood concentration-time curve (AUC) of kagrillitide for up to 24 hours after administration. 0-24hr ) is the area under the total blood concentration-time curve (AUC) total ) may be 20% or less, 10% or less, 5% or less, 0.1 to 20%, 1 to 10%, or 1 to 5%.

[0046] The pharmaceutical composition according to the present invention has an area under the blood concentration-time curve (AUC) of kagrillitide from the day of administration to the day of the dosing interval. 0-QXM ) is the area under the total blood concentration-time curve (AUC) total) in comparison, for humans it may be 70% or less, 65% or less, 60% or less, 20 to 70%, 20 to 65%, 20 to 60%, 25 to 70%, 25 to 65%, 25 to 60%, 30 to 70%, 30 to 65% or 30 to 60%, and for rats it may be 95% or less, 93% or less, 90% or less, 50 to 95%, 50 to 93%, 50 to 90%, 55 to 95%, 55 to 93%, 55 to 90%, 60 to 95%, 60 to 93%, or 60 to 90%.

[0047] In addition, the pharmaceutical composition according to the present invention may be a pharmaceutical composition comprising sustained-release microspheres in which the content of kagrillintide is 12% by weight or more as kagrillintide based on the total weight of the microspheres and the initial release of the drug is less than 10% within 24 hours.

[0048]

[0049] The biodegradable polymers included in the sustained-release microspheres of Kagrilintide included in the pharmaceutical composition according to the present invention are polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA), polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkylcarbonate, polytrimethylenecarbonate (PTMC), A polymer selected from the group consisting of polylactide-co-trimethylenecarbonate (PLTMC), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo polyaminoacid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphoester, polysaccharide and poly(butylene succinate lactide) (PBSLA), the selected polymer A simple mixture of two or more substances,It is selected from the group consisting of a copolymer of the above-mentioned selected polymer and polyethylene glycol (Polyethylenglycol, PEG), and a polymer-sugar complex in which the above-mentioned selected polymer or copolymer is combined with sugar.

[0050] In addition, the biodegradable polymer may be a simple mixture of two or more (i.e., a simple mixture including two or more of the polymers selected from the above), which may include two or more different types of polymers among the non-limiting examples of the polymers, or may be a combination or blend of the same type of polymers, but may be a combination of polymers of the same type having different intrinsic viscosities and / or monomer ratios (e.g., a combination or blend of two or more poly(lactide-co-glycolide) having different intrinsic viscosities), or may be a polymer of the same type having different terminal groups (e.g., a terminal group of an ester or a terminal group of an acid).

[0051] In one specific embodiment, the pharmaceutical composition according to the present invention may include microspheres containing two or more types of the biodegradable polymers described above. In another specific embodiment, the pharmaceutical composition according to the present invention may include two or more types of microspheres each containing one or more types of two or more polymers selected from the biodegradable polymers described above.

[0052] When the biodegradable polymer is polylactide-co-glycolide, the molar ratio of lactide to glycol in the copolymer may be 40:60 to 90:10, 45:55 to 85:15, or 50:50 to 75:25, for example, 45:55, 50:50, 75:25, or 85:15.

[0053] The above biodegradable polymer may be a biodegradable polymer having an intrinsic viscosity of 0.16 to 1.7 dL / g, 0.2 to 1.3 dL / g, or 0.24 to 1.2 dL / g, taking into account factors such as the release characteristics of the drug and the manufacturing process. The above intrinsic viscosity refers to that measured at a concentration of 0.1% (w / v) in chloroform at 25°C using an Ubbelohde viscometer.

[0054] When the intrinsic viscosity of the biodegradable polymer is less than 0.16 dL / g, the molecular weight of the polymer is insufficient, making it difficult to exhibit the sustained-release effect of kagrillintide or a pharmaceutically acceptable salt thereof. When the intrinsic viscosity of the biodegradable polymer exceeds 1.7 dL / g, the release of kagrillintide or a pharmaceutically acceptable salt thereof may be excessively delayed. In addition, when manufacturing microspheres using a polymer with a high intrinsic viscosity, there is a problem that an excessive amount of manufacturing solvent must be used due to the high viscosity of the polymer, and it is difficult to manufacture reproducible microspheres. Examples of commercially available polymers having the above properties include the Resomer® series from Evonik: RG502H, RG503H, RG504H, RG502, RG503, RG504, RG653H, RG752H, RG753H, RG752S, RG755S, RG750S, RG757S, RG858S, R202H, R203H, R205H, R202S, R203S, R205S, R206S, and R207S; PDL 02A, PDL 02, PDL 04, PDL 05, PDLG 7502A, PDLG 7502, PDLG 7507, PDLG 5002A, PDLG 5002; Examples include PDLG 5004A and PDLG 5004.

[0055] The content of the biodegradable polymer in the sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention may be selected as an upper limit of 60 wt% or more, 62 wt% or more, 65 wt% or more, 67 wt% or more, or 70 wt% or more, based on the total weight of the microspheres, and may be selected as a lower limit of 92 wt% or less, 91 wt% or less, 90 wt% or less, 89 wt% or less, or 88 wt% or less, or may be included in a range consisting of a combination of the above upper and lower limits. For example, it may be 60 wt% to 92 wt%, 65 to 90 wt%, or 70 to 88 wt%, but is not limited thereto.

[0056] The content of kagrillintide or a pharmaceutically acceptable salt thereof in the sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention is preferably 8 wt% or more, 12 wt% or more and 14 wt% or more, 37 wt% or less, 35 wt% or less, or 33 wt% or less, 8 wt% to 37 wt%, 8 wt% to 35 wt%, 8 wt% to 33 wt%, 12 wt% to 37 wt%, 12 wt% to 35 wt%, 12 wt% to 33 wt%, 14 wt% to 37 wt%, 14 wt% to 35 wt%, or 14 wt% to 33 wt% as kagrillintide, based on the total weight of the microspheres. If the content of kagrillintide or a pharmaceutically acceptable salt thereof in the microparticles is less than 8 wt% based on kagrillintide, the amount of polymer used compared to the drug may be too high, which may lower the bioavailability of kagrillintide or a pharmaceutically acceptable salt thereof, and if the content is excessively high, the initial release of kagrillintide or a pharmaceutically acceptable salt thereof may be increased, which may be undesirable.

[0057] A pharmaceutical composition comprising the western-type microspheres according to the present invention may contain an early-release inhibitor in the microspheres in an amount less than a certain amount.

[0058] In the present invention, the early-release inhibitor is a substance included in the continuous phase to suppress the rapid release of the active ingredient in the microsphere manufacturing process, and is characterized in that it exists in the microsphere according to the present invention in less than a certain amount. The continuous phase refers to a substance that enables the dispersion phase including the active ingredient such as kagrillintide and a biodegradable polymer to be dispersed when using an emulsion and solvent extraction evaporation method in the method for manufacturing the microspheres, and in the present invention, it may be an aqueous solution including a surfactant, but is not limited thereto. The early-release inhibitor according to the present invention is included in the continuous phase used in this manufacturing method, and can be included in a certain amount in the microsphere according to the present invention after manufacturing accordingly. Matters regarding the continuous phase can be equally applied to matters described in the method for manufacturing the microsphere according to the present invention described below.

[0059] In a specific embodiment, the initial release inhibitor is present in an amount of 10 ppm or more, 20 ppm or more, 30 ppm or more, 40 ppm or more, 50 ppm or more, 100 ppm or more, 150 ppm or more, 200 ppm or more, 250 ppm or more, 200 ppm or more, 300 ppm or more, 350 ppm or more, 400 ppm or more, 450 ppm or more, 500 ppm or more, 550 ppm or more, 600 ppm or more, 650 ppm or more, 700 ppm or more, 750 ppm or more, 800 ppm or more, 850 ppm or more, 900 ppm or more, 950 ppm or more, 1000 ppm or more, 1100 ppm or more, 1200 ppm or more, 1300 ppm or more, based on the total weight of the microparticles. The upper limit may include 1400 ppm or more, or 1500 ppm or more, and the lower limit may include 2000 ppm or less, 1900 ppm or less, 1800 ppm or less, 1700 ppm or less, 1600 ppm or less, 1500 ppm or less, 1400 ppm or less, 1300 ppm or less, 1200 ppm or less, 1100 ppm or less, 1000 ppm or less, 900 ppm or less, 800 ppm or less, 700 ppm or less, 600 ppm or less, or 500 ppm or less, and the range may be comprised of a combination of the above upper and lower limits. For example, the early release inhibitor may be included in an amount of 10 ppm to 2000 ppm, preferably 20 ppm to 1500 ppm, more preferably 20 ppm to 1000 ppm, and most preferably 30 ppm to 500 ppm, relative to the total weight of the microspheres.

[0060]

[0061] In a specific embodiment, the above early release inhibitor may be any substance that, when dissolved in the continuous phase, allows the pH of the continuous phase to be set to 7 or higher.

[0062] The above early release inhibitor may be at least one selected from, but is not limited to, phosphate salts, hydroxide salts, phosphide salts, phosphite salts, carbonate salts, bicarbonate salts, chromate salts, dichromate salts, oxides, oxalate salts, silicate salts, sulfate salts, sulfide salts, sulfite salts, tartrate salts, tetraborate salts, thiosulfate salts, arsenate salts, arsenite salts, citrate salts, ferricyanide salts, and nitride salts of alkali metals, alkaline earth metals, or ammonium.

[0063] In a specific embodiment, the early release inhibitor may be at least one selected from the group consisting of phosphate salts of two or more alkali metals, carbonate salts of one or more alkali metals, and bicarbonate salts of one or more alkali metals, but is not limited thereto.

[0064] In a more specific embodiment, the phosphate salt of the two or more alkali metals means a phosphate salt containing two or more alkali metal ions, for example, disodium phosphate (Na2HPO4) or dipotassium phosphate (K2HPO4), the bicarbonate salt of the one or more alkali metals means a bicarbonate salt containing one or more alkali metal ions, for example, sodium bicarbonate (NaHCO3), and the carbonate salt of the two or more alkali metals means a carbonate salt containing two or more alkali metal ions, for example, sodium carbonate (Na2CO3), and these may be used alone or in combination of two or more.

[0065]

[0066] The microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention have an average particle size of 5 μm to 100 μm, 5 μm to 90 μm, 5 μm to 80 μm, 10 μm to 90 μm, 10 μm to 80 μm, 15 μm to 100 μm, 15 μm to 90 μm, 15 μm to 80 μm, 70 μm to 100 μm, 70 μm to 90 μm, 70 μm to 80 μm, 60 μm to 100 μm, 60 μm to 80 μm, 60 μm to 70 μm, 20 μm to 90 μm, 20 μm to 70 μm, 20 μm to 60 μm, 30 μm to It is preferable to have a uniform particle distribution of 80 μm, 30 μm to 60 μm, 40 μm to 70 μm, 40 μm to 50 μm, 30 μm to 40 μm, 20 μm to 30 μm, 5 μm to 30 μm, 5 μm to 20 μm, 10 μm to 20 μm, or 5 μm to 10 μm. The term “average particle size” used in the present invention refers to a particle size corresponding to 50% of the volume % in a particle size distribution curve, which means median diameter and is expressed as D50 or D(v, 0.5).

[0067] If the average particle size of the microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor is less than 5 μm, the release of the drug kagrillintide or a pharmaceutically acceptable salt thereof from the microspheres may be too rapid, which may be undesirable. If the average particle size exceeds 100 μm, the injection needle may become too thick during human administration, which may be undesirable because it may cause pain during injection or the drug may leak out of the injection site after injection.

[0068] It is preferable that the microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor of the present invention have a uniform particle distribution. Microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor having a uniform particle distribution have a smaller deviation during injection and can be administered in a more accurate amount compared to microspheres having a non-uniform particle distribution. It is preferable that the microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor of the present invention have a span value of 1.5 or less. More preferably, the span value is 1.2 or less. More specifically, the span value may be 1.5 or less, 1.2 or less, 0.1 to 1.5, 0.3 to 1.5, 0.5 to 1.5, 0.1 to 1.0, 0.4 to 1.0, 0.6 to 1.0, 0.2 to 0.8, or 0.4 to 0.8. The term “span value” used in the present invention is an indicator representing the uniformity of the particle size of the microspheres, and means a value obtained by the formula of Span value = (Dv0.9-Dv0.1) / Dv0.5. Here, Dv0.1 means the particle size corresponding to 10% of the volume% in the particle size distribution curve of the microspheres, Dv0.5 means the particle size corresponding to 50% of the volume% in the particle size distribution curve of the microspheres, and Dv0.9 means the particle size corresponding to 90% of the volume% in the particle size distribution curve of the microspheres.

[0069] The sustained-release microspheres of the present invention containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor are administered by an injection route, for example, subcutaneous injection, and are particularly preferred to release kagrillintide for a relatively long period of time because they can be self-administered. Preferably, the sustained-release microspheres in the pharmaceutical composition according to the present invention can release kagrillintide or a pharmaceutically acceptable salt thereof for, but not limited to, 1 month or more, 2 months or more, 3 months or more, 1 month to 2 months, 1 month to 3 months, 1 month to 4 months, 1 month to 5 months, 1 month to 6 months, 2 months to 6 months, 2 months to 5 months, 2 months to 4 months, 2 months to 3 months, 3 months to 5 months, or 3 months to 4 months. In addition, the sustained-release microspheres of the present invention containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor are preferably released within 24 hours when administered in vivo at a release rate (release amount) of kagrillintide or a pharmaceutically acceptable salt thereof with an upper limit of less than 10%, less than 15%, or less than 20%, or at a release rate of 0.001% or more, 0.01% or more, 0.1% or more, or 1% or more, or at a release rate in a range of a combination of the above upper and lower limits, although the release pattern is not particularly limited.

[0070] In addition, in the pharmaceutical composition of the present invention, the total amount (total weight) of the sustained-release microsphere formulation comprising the above-described caprylintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor is 20 to 1000 mg, 20 to 800 mg, 20 to 600 mg, 20 to 400 mg, 20 to 200 mg, 20 to 100 mg, 30 to 1000 mg, 30 to 800 mg, 30 to 600 mg, 30 to 400 mg, 30 to 200 mg, 30 to 100 mg, 40 to 1000 mg, 40 to 800 mg, 40 to 600 mg, 40 to 400 mg, 40 to 200 mg, 40 to It may be 100 mg, 50 mg to 1000 mg, 50 mg to 800 mg, 50 mg to 600 mg, 50 mg to 400 mg, 50 mg to 200 mg, or 50 mg to 100 mg. By including sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor in the composition within the above range, the composition according to the present invention not only minimizes inflammatory reactions at the site of administration, but also has the advantage of enabling self-administration by the patient.

[0071] The microspheres comprising caprylintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor included in the composition of the present invention may further comprise a release-controlling agent. Examples of the material used as the release-controlling agent include, but are not limited to, one or more selected from butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, nonadecylic acid, behenic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, napadicylic acid, naphthalene sulfonic acid, and pamoic acid. Preferably, the release controlling agent may be, but is not limited to, hydroxynaphthoic acid, napadiylic acid or pamoic acid.

[0072]

[0073] The pharmaceutical composition comprising microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention can be formulated into various forms of preparations, and for example, can be a formulation of a known parenteral administration preparation. Accordingly, the pharmaceutical composition according to the present invention can further comprise a thickener, a stabilizer, an isotonic agent, a surfactant, an excipient, and / or a carrier in addition to the kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor. Usable isotonic agents can be water-soluble excipients or sugars such as mannitol, sucrose, sorbitol, trehalose, lactose, sodium chloride, etc., and examples of thickeners include sodium carmellose, sodium carboxymethyl cellulose, povidone, etc. In addition, stabilizers can be used such as sodium monohydrogen phosphate, anhydrous citric acid, sodium hydroxide, sodium chloride, etc.

[0074]

[0075] The pharmaceutical composition according to the present invention may be administered in a therapeutically effective amount of kagrillintide, for example, an amount effective for treating diabetes, specifically type 2 diabetes, beta-cell dysfunction, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, or neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. The therapeutically effective amount of kagrillintide may be assessed by a physician. The pharmaceutical composition according to the present invention comprising kagrillintide may be administered once a month to once a quarter. In some embodiments, the monthly dosage of the composition according to the present invention is, based on kagrilintide, 1 mg to 100 mg, 1 mg to 80 mg, 1 mg to 60 mg, 1 mg to 30 mg, 1 mg to 20 mg, 1 mg to 10 mg, 1 mg to 8 mg, 1 mg to 6 mg, 2 mg to 100 mg, 2 mg to 80 mg, 2 mg to 60 mg, 2 mg to 30 mg, 2 mg to 20 mg, 2 mg to 10 mg, 2 mg to 8 mg, 2 mg to 6 mg, 4 mg to 100 mg, 4 mg to 80 mg, 4 mg to 60 mg, 4 mg to 30 mg, 4 mg to 20 mg, 8 mg to 100 mg, 8 mg to 80 mg, 8 mg to 60 mg, 8 mg to The dosage may be 30 mg, 10 mg to 100 mg, 10 mg to 80 mg, 10 mg to 60 mg, 10 mg to 30 mg, 20 mg to 100 mg, 20 mg to 80 mg, 20 mg to 60 mg, 20 mg to 30 mg. The pharmaceutical composition according to the present invention comprising microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention may be administered parenterally, for example, via subcutaneous injection.The pharmaceutical composition according to the present invention may be composed of a drug part containing microparticles including kagrillitide or a pharmaceutically acceptable salt thereof and an early-release inhibitor, and a solvent part used to suspend the microparticles, and may be in the form of a dual-chamber syringe having the drug part in one chamber and the solvent part in the other chamber, or a prefilled syringe in which the drug part is suspended in the solvent part. When the pharmaceutical composition is composed in the form of a prefilled syringe in which the drug part is suspended in the solvent part, the solvent part used may be an injectable oil including medium-chain oil, mineral oil, etc.

[0076] In a specific embodiment, the sustained-release microspheres containing caprylintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor included in the pharmaceutical composition according to the present invention have a high drug content relative to the content of the microspheres, while suppressing the initial excessive release of the drug that may cause fatal side effects and exhibiting high bioavailability to sufficiently exhibit the efficacy of an amylin analog for a desired period of time, and are useful for the prevention or treatment of diabetes, specifically type 2 diabetes, beta-cell dysfunction, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, or Alzheimer's disease.

[0077] In another aspect, the present invention provides a pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta-cell dysfunction, obesity, non-alcoholic steatohepatitis, Alzheimer's disease or neurodegenerative diseases, comprising sustained-release microspheres containing caprylintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor and semaglutide sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a biodegradable polymer.

[0078] The aforementioned matters may be applied to sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor in the pharmaceutical composition.

[0079] In a specific embodiment, the microspheres comprising semaglutide included in the pharmaceutical composition according to the present invention provide a pharmaceutical composition comprising semaglutide sustained-release microspheres having a semaglutide content of at least 8 wt% as semaglutide based on the total weight of the microspheres.

[0080] The content of semaglutide or a pharmaceutically acceptable salt thereof in the sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof according to the present invention is preferably 8 wt% or more, 12 wt% or more and 14 wt% or more, 37 wt% or less, 35 wt% or less, or 33 wt% or less, 8 wt% to 37 wt%, 8 wt% to 35 wt%, 8 wt% to 33 wt%, 12 wt% to 37 wt%, 12 wt% to 35 wt%, 12 wt% to 33 wt%, 14 wt% to 37 wt%, 14 wt% to 35 wt%, or 14 wt% to 33 wt% as semaglutide based on the total weight of the microspheres.

[0081]

[0082] In another aspect, the present invention provides a pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta-cell dysfunction, obesity, non-alcoholic steatohepatitis, Alzheimer's disease or neurodegenerative diseases, comprising sustained-release microspheres simultaneously containing capriglintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof and an early-release inhibitor.

[0083] In a specific embodiment, the sustained-release microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof, semaglutide or a pharmaceutically acceptable salt thereof, and an early-release inhibitor included in the pharmaceutical composition according to the present invention provide a pharmaceutical composition comprising sustained-release microspheres comprising kagrillintide and semaglutide, wherein the content of kagrillintide and semaglutide is each 4 wt% or more based on the total weight of the microspheres, and the early-release inhibitor is 10 ppm to 2000 ppm based on the total weight of the microspheres.

[0084] The content of each of capgrilintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof in the sustained-release microspheres containing capgrilintide or a pharmaceutically acceptable salt thereof, semaglutide or a pharmaceutically acceptable salt thereof, and an early-release inhibitor according to the present invention is 4 wt% or more, 6 wt% or more, 7 wt% or more, 20 wt% or less, 18.5 wt% or less, 17.5 wt% or less, or 16.5 wt% or less, 4 wt% to 20 wt%, 4 wt% to 18.5 wt%, 4 wt% to 17.5 wt%, or 4 wt% to 16.5 wt%, 6 wt% to 20 wt%, 6 wt% to 18.5 wt%, 6 wt% to 17.5 wt%, based on the total weight of the microspheres. Or, it is preferably 6 wt% to 16.5 wt%, 7 wt% to 20 wt%, 7 wt% to 18.5 wt%, 7 wt% to 17.5 wt% or 7 wt% to 16.5 wt%.

[0085] In addition, the content of the early-release inhibitor in the sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof, semaglutide or a pharmaceutically acceptable salt thereof, and an early-release inhibitor according to the present invention is the same as that for the kagrillintide microspheres described above.

[0086]

[0087] In addition, all matters related to microspheres, such as biodegradable polymers and their contents, can be applied as is to the matters related to Kagrilintide microspheres.

[0088] The maximum efficacy achievable with the GLP-1 receptor agonist semaglutide is limited by tolerability. As the dose is increased, adverse reactions such as nausea and vomiting become increasingly prominent. Amylin receptor agonist therapy is limited by tolerability (and similar adverse reactions such as nausea and vomiting) in much the same way as GLP-1 receptor agonist therapy, and there is hope for extending the action of amylin, suggesting the need for combining the amylin receptor agonist cagrilintide with the GLP-1 receptor agonist semaglutide.

[0089] However, it is known that semaglutide and capgrilintide cannot be formulated in the same pharmaceutical formulation. To ensure both chemical and physical stability, semaglutide must be formulated at a pH of 7-8, for example, approximately pH 7.4. To ensure both chemical and physical stability, capgrilintide must be formulated at a pH of 3.5-4.5, for example, approximately pH 4.0. Because of these significant formulation differences, the two pharmaceutical formulations containing semaglutide and capgrilintide must be stored separately until administration. Therefore, co-administration requires separate administration or mixing. Since capgrilintide is administered via subcutaneous injection, separate administration can cause additional pain and discomfort for patients. Therefore, administering capgrilintide via a single injection, rather than two, would improve treatment compliance and ultimately enhance the desired pharmacological outcome. Additionally, administering capric acid and semaglutide as a single injection rather than separately may affect the efficacy of these active pharmaceutical ingredients.

[0090] In relation to this, there have been attempts to develop dual-chamber devices containing these drugs in the past, but there was a problem that the drugs were not stabilized and particles were generated during storage in the dual-chamber device.

[0091] However, in the case of the present invention, when capillary ligand and semaglutide are simultaneously encapsulated in microspheres or microspheres encapsulating each component are combined, the stability of the drug is secured while the need for separate pH adjustment and buffer selection as mentioned above is eliminated, so that not only the cost problem arising from manufacturing separate formulations but also the inconvenience of having to mix them immediately before administration can be eliminated.

[0092]

[0093] In another aspect, the present invention provides a method for preparing sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof as an active ingredient; a biodegradable polymer; and an early-release inhibitor.

[0094] In another aspect, the present invention provides a method for preparing sustained-release microspheres having significantly suppressed initial release despite containing a high content of kagrillintide or a pharmaceutically acceptable salt thereof.

[0095] Hereinafter, the method for manufacturing the western-type microparticle injection of the present invention will be described in detail.

[0096] The western-type microspheres according to the present invention can be manufactured using, for example, “emulsion solvent extraction and evaporation method,” but the manufacturing method is not limited thereto.

[0097] As one embodiment of a specific manufacturing method, the present invention provides a method for manufacturing a sustained-release microsphere with an O / W emulsion comprising the following steps:

[0098] (a) a step of preparing an oil phase (O phase) in which (i) kagrillintide or a pharmaceutically acceptable salt thereof or (ii) kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof and one or more biodegradable polymers are dissolved in an organic solvent as an active ingredient;

[0099] (b) a step of preparing a continuous phase (W phase) by adding an early release inhibitor to an aqueous solution containing a surfactant, and adding the dispersed phase of step (a) to prepare a dispersed phase in an emulsion state;

[0100] (c) a step of extracting and evaporating an organic solvent from the dispersed phase of the emulsion state prepared in step (b) into a continuous phase (W phase) to form microspheres;

[0101] (d) Step of recovering the microspheres.

[0102] When kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof are used as active ingredients in the above step (a), co-encapsulated microspheres containing kagrillintide and semaglutide simultaneously can be prepared.

[0103]

[0104] In another aspect, the present invention provides a method for preparing a western-type microsphere as a W / O / W emulsion comprising the following steps:

[0105] (a') a step of preparing a water phase (W1 phase) by dissolving (i) kagrillintide or a pharmaceutically acceptable salt thereof or (ii) kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof as an active ingredient in an aqueous solution, and preparing an oil phase (O phase) by dissolving one or more biodegradable polymers in an organic solvent, and preparing a first W1 / O emulsion by mixing the water phase (W1 phase) and the oil phase (O phase) as a dispersion phase;

[0106] (b') A step of preparing a continuous phase (W2 phase) by adding an early release inhibitor to an aqueous solution containing a surfactant, and adding the dispersed phase of step (a') to prepare a dispersed phase in a secondary W1 / O / W2 emulsion state;

[0107] (c') A step of forming microspheres by extracting and / or evaporating an organic solvent from the dispersed phase of the secondary W1 / O / W2 emulsion state of step (b') into a continuous phase (W2 phase);

[0108] (d') Step of recovering the microspheres.

[0109]

[0110] When kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof are used as active ingredients in the above step (a'), co-encapsulated microspheres containing kagrillintide and semaglutide simultaneously can be prepared.

[0111]

[0112] In the case where two types of drugs are included in the preparation of the aqueous phase in the above step (a'), the two types of drugs may be dissolved simultaneously in one aqueous solution to prepare the aqueous phase (W phase), or the two types of drugs may be dissolved in separate aqueous solutions to prepare the aqueous phase a (W1a phase) and aqueous phase b (W1b phase). When the aqueous phase a (W1a phase) and aqueous phase b (W1b phase) are prepared, they may be mixed simultaneously with the oil phase (O phase) or mixed sequentially to prepare the W1 / O emulsion as the dispersed phase.

[0113]

[0114] In another aspect, the pH of the continuous phase used in the above manufacturing methods may be 7 or higher.

[0115] In the manufacture of sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention, the following biodegradable polymers may be used to suppress excessive early release of kagrillintide or a pharmaceutically acceptable salt thereof while containing a high amount of kagrillintide or a pharmaceutically acceptable salt thereof relative to the weight of the microspheres, and to release at a constant concentration for a desired long period of time, for example, for 1 month or more, 3 months or more, 1 month to 2 months, 1 month to 3 months, 1 month to 4 months, 1 month to 5 months, 1 month to 6 months, 2 months to 6 months, 2 months to 5 months, 2 months to 4 months, 2 months to 3 months, 3 months to 5 months, or 3 months to 4 months. Specifically, polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA), polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkylcarbonate, polytrimethylenecarbonate (PTMC), polylactide-co-trimethylenecarbonate (PLTMC), polyhydroxybutyric acid (Polyhydroxybutyric acid) Polyhydroxybutyrate-co-hydroxyvalerate (PHBV),It is preferable to use at least one polymer, preferably at least two polymers, selected from the group consisting of a polymer selected from the group consisting of polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo polyaminoacid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphoester, polysaccharide and poly(butylene succinate lactide) (PBSLA), a simple mixture of two or more of the above-mentioned polymers, a copolymer of the above-mentioned polymer and polyethylenglycol (PEG), and a polymer-sugar complex in which the above-mentioned polymer or copolymer is bound to a sugar. In a specific embodiment, in the manufacturing method according to the present invention, poly(lactide-co-glycolide) and / or polylactide polymer may be used as the biodegradable polymer.

[0116] In the present invention, the two or more different biodegradable polymers may include two or more polymers having different repeating units constituting the polymers, and two or more polymers having different molar ratios of the repeating units when including two or more repeating units. As an example, the microspheres may be a mixture of microspheres including poly-lactide-co-glycolide and microspheres including polylactide polymer, or microspheres including both poly-lactide-co-glycolide and polylactide polymer.

[0117] In addition, as a specific example, when there are two types of biodegradable polymers that are different from each other, the content ratio of these biodegradable polymers may be, but is not limited to, a weight ratio of 0.5:10 to 10:0.5, 0.5:8 to 8:0.5, 1:10 to 10:1, 1:4 to 4:1, 1:3 to 3:1, or 1:2 to 2:1.

[0118] Additionally, unless otherwise defined, the content of the biodegradable polymer described in the microspheres according to the present invention may be applied as is.

[0119] In a more specific embodiment, when polylactide-co-glycolide is used as two or more biodegradable polymers to prepare sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor according to the present invention, at least one biodegradable polymer having an intrinsic viscosity of 0.16 dL / g to 0.45 dL / g may be included.

[0120] For the above polylactide-co-glycolide, the molar ratio of lactide to glycol in the copolymer may be 40:60 to 90:10, 45:55 to 85:15, or 50:50 to 75:25, for example, 45:55, 50:50, 75:25, or 85:15.

[0121] In step (a) or (a') of the above specific manufacturing method, the organic solvent used to dissolve at least one biodegradable polymer is at least one organic solvent. In addition, the organic solvent may be a mixed organic solvent obtained by mixing two or more organic solvents. In a specific embodiment, the mixed solvent may be a mixed solvent of an organic solvent that is miscible with water and an organic solvent that is immiscible with water. In this case, it is preferable to use at least 50% (v / v) or more, 60% (v / v) or more, 50 to 99.9% (v / v) or more, 50 to 90% (v / v) or more, 50 to 80% (v / v) or more, 50 to 70% (v / v) or more, 60 to 90% (v / v) or more, or 60 to 80% (v / v) or more of the organic solvent that is immiscible with water. By utilizing the property of the organic solvent not to be miscible with water, an emulsion can be formed by homogeneously mixing the dispersed phase in the continuous phase containing the surfactant in step (b) or (b') described below. The type of the organic solvent that dissolves one or more biodegradable polymers is not particularly limited, but preferably, a mixed solvent of one 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, more preferably, one solvent selected from dichloromethane and ethyl acetate and one or more organic solvents selected from dimethyl sulfoxide, N-methylpyrrolidone, methyl alcohol, and acetic acid can be used. In one embodiment, a mixture of dichloromethane and acetic acid (glacial acetic acid) may be used as the organic solvent.

[0122] The method for homogeneously mixing the dispersed phase and the continuous phase containing the surfactant in the above step (b) or (b') is not particularly limited, but may be performed using a high-speed stirrer, an inline mixer, a membrane emulsion method, a microfluidic emulsion method, an ultrasonic mixer, or a static mixer, alone or in combination of two or more. When forming an emulsion using a high-speed stirrer, an inline mixer, an ultrasonic mixer, or a static mixer, it is difficult to obtain a uniform emulsion, so it is preferable to additionally perform a sieving process, etc. between steps (c) and (d) described below, or between steps (c') and (d').

[0123] The type of surfactant used in the above step (b) or (b') is not particularly limited, and any surfactant that can help the dispersed phase form a stable droplet dispersed phase within the continuous phase may be used. The surfactant may be selected from the group consisting of polyvinyl alcohol, methylcellulose, polyvinylpyrrolidone, carboxymethylcellulose, lecithin, gelatin, polyoxyethylene sorbitan fatty acid ester, and polyoxyethylene castor oil derivatives, and may be used alone or in combination of two or more.

[0124] In the above step (b) or (b'), the content of the surfactant in the continuous phase containing the surfactant may be 0.01 w / v% to 20 w / v%, preferably 0.03 w / v% to 18 w / v%, 0.05 w / v% to 15 w / v%, 0.07 w / v% to 10 w / v% or 0.1 w / v% to 5 w / v%, based on the total volume of the continuous phase including the surfactant. When the content of the surfactant is less than 0.01 w / v%, a dispersed phase or emulsion in the form of droplets may not be formed in the continuous phase, and when the content of the surfactant exceeds 20 w / v%, after fine particles are formed in the continuous phase due to an excessive amount of surfactant, it may be difficult to remove the surfactant.

[0125] The continuous phase used in the above step (b) or (b') may be an aqueous solution containing a surfactant and an early-release inhibitor. Furthermore, the continuous phase may be added to the aqueous solution containing the surfactant to reach the final concentration of the early-release inhibitor and used. Depending on the use of the early-release inhibitor, the early-release of microparticles encapsulated with a high concentration of the drug can be controlled, and a substance forming a multivalent anion can be used as the early-release inhibitor.

[0126] The type of the above early-release inhibitor is not particularly limited, and any basic salt that can be dissolved in a continuous phase and maintained at a pH of 7.0 or higher can be used. The above early-release inhibitor may be at least one substance selected from among phosphate salts, hydroxide salts, phosphide salts, phosphite salts, carbonate salts, bicarbonate salts, chromate salts, dichromate salts, oxides, oxalate salts, silicate salts, sulfate salts, sulfide salts, sulfite salts, tartrate salts, tetraborate salts, thiosulfate salts, arsenate salts, arsenite salts, citrate salts, ferricyanide salts, and nitride salts of alkali metals, alkaline earth metals, or ammonium.

[0127] In a specific embodiment, the early release inhibitor may be at least one selected from the group consisting of phosphate salts of two or more alkali metals, bicarbonate salts of one or more alkali metals, and carbonate salts of one or more alkali metals, but is not limited thereto.

[0128] In a more specific embodiment, the phosphate salts of the two or more alkali metals are disodium phosphate (Na2HPO4) or dipotassium phosphate (K2HPO4), the bicarbonate salt of the one or more alkali metals is sodium bicarbonate (NaHCO3), and the carbonate salt of the one or more alkali metals is sodium carbonate (Na2CO3). These may be used alone or in combination of two or more.

[0129]

[0130] The final concentration of the early release inhibitor in the above continuous phase may be 0.1 to 5.0 (w / v)%, preferably 0.1 to 4.0 (w / v)%, more preferably 0.2 to 4.0 (w / v)%, still more preferably 0.3 to 3.5 (w / v)%, particularly preferably 0.4 to 3.0 (w / v)%, and particularly more preferably 0.5 to 2.5 (w / v)%.

[0131] If the content of the initial release inhibitor exceeds 5.0 w / v%, the emulsion droplets may burst during the manufacture of microspheres. Conversely, if it is lower than 0.1 w / v%, the initial release of microspheres containing a high content of kagrillintide may not be sufficiently suppressed.

[0132] When the above early release inhibitor is included in the continuous phase during the manufacture of microspheres, the early release from microspheres containing 12 (w / w)% or more of kagrillintide can be reduced preferably to 10% or less, more preferably to 8% or less, and most preferably to 5% or less.

[0133] The continuous phase used in the above step (b) or (b') may further include at least one selected from the group consisting of methyl alcohol, ethyl alcohol, propyl alcohol, and ethyl acetate in order to control the extraction rate of the organic solvent from the dispersed phase in the emulsion state.

[0134] The early release inhibitor may be added at the time of preparing the continuous phase in the above step (b) or (b'), and the early release inhibitor may also be added at any time during the supply of the continuous phase in the above step (c) or (c'), and the method of addition is not limited.

[0135] Additionally, the pH of the continuous phase may be, but is not limited to, 7.0 or more, 7.2 or more, 7.4 or more, 8.0 or more, 8.5 or more, 9.0 or more, 7.0 to 9.0, or 7.0 to 8.5. When the pH of the continuous phase is adjusted within the above range, the bioavailability of microspheres comprising kagrillitide or a pharmaceutically acceptable salt thereof may be further increased.

[0136] In the above step (c) or (c'), when the emulsion including the dispersed phase in the form of droplets and the continuous phase containing the surfactant is maintained or stirred at a temperature below the boiling point of the organic solvent for a certain period of time, for example, 2 to 48 hours, the organic solvent can be extracted as a continuous phase from the dispersed phase, which is the droplet-shaped kagrillintide or a pharmaceutically acceptable salt thereof and the polymer solution. A portion of the organic solvent extracted as a continuous phase can be evaporated from the surface. As the organic solvent is extracted and evaporated from the droplet-shaped kagrillintide or a pharmaceutically acceptable salt thereof and the polymer solution, the droplet-shaped dispersed phase can be solidified to form microspheres.

[0137] In order to further efficiently remove the organic solvent in step (c) or (c'), the temperature of the continuous phase may be heated for a certain period of time. The heating temperature is not limited and can be appropriately adjusted by a person skilled in the art depending on the organic solvent used. For example, when dichloromethane is used as the organic solvent, the heating may be applied so as to maintain the temperature at 30°C or higher, 40°C or higher, 45°C or higher, 30 to 50°C, 40 to 50°C, or 45°C.

[0138] In the above step (d) or (d'), the method for recovering microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor can be performed using various known techniques, for example, methods such as filtration or centrifugation can be used.

[0139] Between the above steps (c) and (d) or between steps (c') and (d'), the remaining surfactant can be removed through filtration and washing, and then filtered again to recover the microspheres.

[0140] The washing step to remove the remaining surfactant can be typically performed using water, and the washing step can be repeated several times.

[0141] Additionally, as described above, a sieving process may be additionally employed between steps (c) and (d) or between steps (c') and (d') to obtain uniform microspheres. The sieving process may be performed using known techniques, and microspheres of uniform size may be obtained by filtering out small and large particles using sieves of different sizes.

[0142] The manufacturing method of the present invention can obtain finally dried microparticles by drying the obtained microparticles using a conventional drying method after the step (d) or step (d') or after the filtering and washing step.

[0143] In addition to the above, all matters including kagrillintide, early-release inhibitor, biodegradable polymer and their contents that are not separately defined may be applied as defined in the above pharmaceutical composition.

[0144] According to the manufacturing method of the present invention, a sustained-release microsphere injection containing kagrillintide or a pharmaceutically acceptable salt thereof, which has high bioavailability and maintains an effective concentration for a desired period of time without abrupt, transient release of the drug, can be manufactured. In addition, a sustained-release microsphere injection containing kagrillintide or a pharmaceutically acceptable salt thereof in uniform particles with good dosing capacity and an early-release inhibitor can be manufactured.

[0145]

[0146] In another aspect, the present invention provides a method for preparing sustained-release microspheres containing kagrilintide or a pharmaceutically acceptable salt thereof, and semaglutide or a pharmaceutically acceptable salt thereof and an early-release inhibitor.

[0147] In another aspect, the present invention provides a method for preparing sustained-release microspheres having significantly suppressed initial release despite containing a high content of caprylintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof.

[0148] A specific description of a method for preparing a sustained-release microsphere injection containing kagrillintide or a pharmaceutically acceptable salt thereof, semaglutide or a pharmaceutically acceptable salt thereof, and an early-release inhibitor of the present invention is the same as the method for preparing sustained-release microspheres containing kagrillintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor described above, except that semaglutide or a pharmaceutically acceptable salt thereof is added as an active ingredient.

[0149] The sustained-release microsphere injection containing kagrillintide or a pharmaceutically acceptable salt thereof, semaglutide or a pharmaceutically acceptable salt thereof, and an early-release inhibitor according to the present invention can be manufactured using, for example, “emulsion solvent extraction and evaporation method,” but the manufacturing method is not limited thereto.

[0150] The sustained-release microspheres according to the present invention may be blended by including two or more types of drug microspheres containing the same drug but differing in at least one of their compositions and manufacturing conditions, or may be blended by including two or more types of drug microspheres containing different drugs. In the case of blending microspheres containing two or more types of drug microspheres, the purpose of blending microspheres may be to optimize the release profile of the drug, control the release period, etc.

[0151] Specifically, the different compositions and manufacturing conditions may be at least one selected from the group consisting of, but are not limited to, the amount of drug used, the type of polymer, the particle size distribution (e.g., average particle size), circularity, the amount of polymer used, the type and amount of dispersed phase solvent, the type and amount of co-solvent, the type and amount of continuous phase, the solidification temperature and time, and the theoretical content of the drug.

[0152] The above microsphere blending may be, for example, mixing at least one drug microsphere having different compositions and manufacturing conditions at a specific ratio. Specifically, when blending microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof and microspheres comprising semaglutide or a pharmaceutically acceptable salt thereof, the mixing ratio (weight ratio or ratio of the number of microspheres) may be appropriately adjusted in consideration of the bioavailability of the active ingredient (i.e., kagrillintide or semaglutide) of each microsphere. For example, the weight ratio or number ratio of microspheres comprising capriglintide or a pharmaceutically acceptable salt thereof to microspheres comprising semaglutide or a pharmaceutically acceptable salt thereof may be mixed at a ratio of 1:100, 1:70, 1:50, 1:30, 1:10, 1:5, 1:2, 1:1, 2:1, 5:1, 10:1, 30:1, 50:1, 70:1 or 100:1.

[0153] As a specific example, the two different drug microspheres may be drug microspheres having different components and composition ratios (hereinafter, drug microspheres having different compositions) and / or drug microspheres having different drug release characteristics, and for example, at least one selected from the group consisting of the type of polymer of the microsphere, the polymer content, the drug content, etc. may be different. The type of polymer of the microspheres may be different in at least one selected from the group consisting of the repeating unit of the polymer, the terminal group of the polymer, the molecular weight of the polymer, the intrinsic degree of the polymer, etc.

[0154] The above microsphere blending can be achieved by separately manufacturing and mixing the microspheres, or by manufacturing them in a single manufacturing process. Specifically, an O / W emulsion method for manufacturing the microsphere blending in a single manufacturing process can be produced by, as an example, including the following steps 1 to 4.

[0155] A step of separately preparing two or more types of dispersed solutions by dissolving biodegradable polymers and drugs having different compositions in organic solvents (step 1);

[0156] A step of forming an emulsion of two or more dispersed phases by injecting two or more types of dispersed phases prepared in the above step 1 into an aqueous solution (continuous phase) containing a surfactant (step 2);

[0157] A step of forming microspheres by extracting and evaporating the organic solvent from the dispersed phase emulsion prepared in step 2 to the continuous phase (step 3); and

[0158] It may include a step of recovering the microspheres (step 4) after the above step 3.

[0159] Specifically, a W / O / W emulsion method for manufacturing microparticle blending in one manufacturing process can be manufactured by including the following steps 1 to 4 as an example.

[0160] A step (step 1) of preparing a water phase (W1 phase) in which a drug is dissolved in an aqueous solution, preparing two or more oil phases (O phase) by dissolving biodegradable polymers having different compositions in organic solvents, and separately preparing a dispersed phase solution by mixing the water phase (W1 phase) and two or more oil phases (O phase);

[0161] A step of forming an emulsion of two or more dispersed phases by injecting two or more dispersed phases prepared in the above step 1 into an aqueous solution (W2 phase, continuous phase) containing a surfactant (step 2);

[0162] A step (step 3) of forming microspheres by extracting and evaporating the organic solvent from the dispersed emulsion prepared in step 2 toward the continuous phase (W2 phase); and

[0163] It may include a step of recovering the microspheres (step 4) after the above step 3.

[0164] [Example]

[0165] Hereinafter, the present invention will be described in more detail with reference to the following manufacturing examples. However, the following manufacturing examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following manufacturing examples.

[0166]

[0167] Examples 1-1 to 1-7 (O / W formulation): Preparation of biodegradable polymer microspheres containing kagrilintide and an early-release inhibitor (difference in drug target loading%)

[0168]

[0169] Kagrilintide (manufacturer: Zhejiang Peptides Biotech Co., Ltd, China) was used as the drug, and biodegradable polymers RG653H and RG753H (manufacturer: Evonik, Germany) were used at a weight ratio of 1:1 and weighed so that the batch size was 0.5 g. The drug and polymer were homogeneously dissolved in a mixed solvent of dichloromethane (DCM) solvent and glacial acetic acid co-solvent to prepare an oil phase (O phase) as the dispersed phase.

[0170] Na2HPO4 (Disodium phosphate) was added as an initial release inhibitor to a 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8–5.8 mPa.s) aqueous solution as a continuous phase (W phase) to a final concentration of 2.0 (w / v)%.

[0171] The continuous phase was connected to an emulsifying device equipped with a 40 μm diameter porous membrane, and the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing biodegradable polymer microdroplets containing kagrillintide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm. The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase was injected, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the temperature was filtered to remove the residual polyvinyl alcohol with triple-distilled water, and freeze-dried.

[0172]

[0173] Example No. T / L % Polymer type Polymer amount (g) Kagrilintide amount (g) DCM amount (g) Glacial acetic acid amount (g) Early release inhibitor and concentration 1-115 RG653H:RG753H = 1:10.425 0.075 5.21.4252 (w / v)% Na2HPO4 1-220 RG653H:RG753H = 1:10.40 0.100 4.9 1.60 92 (w / v)% Na2HPO4 1-325 RG653H:RG753H = 1:10.375 0.125 4.6 1.7272 (w / v)% Na2HPO41-430RG653H:RG753H=1:10.3500.1506.71.7022(w / v)% Na2HPO41-535RG653H:RG753H=1:10.3250.1756.21.9852(w / v)% Na2HPO41-640RG653H:RG753H=1:10.3000.2005.71.8412(w / v)% Na2HPO41-750RG653H:RG753H=1:10.2500.2504.81.7342(w / v)% Na2HPO4

[0174] * T / L: Target Loading

[0175] Examples 2-1 to 2-4 (O / W preparation): Preparation of biodegradable polymer microspheres containing kagrillintide and an early-release inhibitor (differences in the type of early-release inhibitor)

[0176] Kagrilintide (manufacturer: Zhejiang Peptides Biotech Co., Ltd, China) was used as the drug, and biodegradable polymers RG653H and RG753H (manufacturer: Evonik, Germany) were used at a weight ratio of 1:1 and weighed so that the batch size was 0.5 g (target loading 15%). The drug and polymer were homogeneously dissolved in a mixed solvent of dichloromethane (DCM) solvent and glacial acetic acid co-solvent to prepare an oil phase (O phase) as the dispersed phase.

[0177] As a continuous phase (W phase), 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8 to 5.8 mPa.s) aqueous solution was used, and K2HPO4 (Dipotassium phosphate), Na2CO3 (Sodium carbonate), NaHCO3 (Sodium bicarbonate), or (NH4)2HPO4 (Diammonium phosphate) was added as an early release inhibitor to a final concentration of 2.0 (w / v) %.

[0178] After connecting the continuous phase to an emulsifying device equipped with a 40 μm diameter porous membrane, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing biodegradable polymer microdroplets containing kagrillintide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm. The temperature of the preparation vessel was maintained at 25°C, and after the injection of the dispersed phase, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the temperature was filtered to remove the residual polyvinyl alcohol with triple-distilled water, and freeze-dried.

[0179]

[0180] Example No. Polymer type Polymer amount (g) Kagrilintide amount (g) DCM amount (g) Glacial acetic acid amount (g) Early release inhibitor and concentration 2-1RG653H:RG753H=1:10.4250.0755.21.4252(w / v)% K2HPO4 2-2RG653H:RG753H=1:10.4250.0755.21.4252(w / v)% Na2CO3 2-3RG653H:RG753H=1:10.4250.0755.21.4252(w / v)% NaHCO3 2-4RG653H:RG753H=1:10.4250.0755.21.4252(w / v)% (NH4)2HPO4

[0181] Examples 3-1 to 3-4 (O / W preparation): Preparation of biodegradable polymer microspheres containing kagrilintide and an early-release inhibitor (differences in polymer types)

[0182] Kagrilintide (manufacturer: Zhejiang Peptides Biotech Co., Ltd, China) was used as the drug, and Rg503H, RG653H, RG753H, or R203H (manufacturer: Evonik, Germany) was used as the biodegradable polymer, and the batch size was weighed to be 0.5 g (target loading 15%). The drug and polymer were homogeneously dissolved in a mixed solvent of dichloromethane solvent and glacial acetic acid co-solvent, and an oil phase (O phase) was prepared as the dispersed phase.

[0183] Na2HPO4 (Disodium phosphate) was added as an initial release inhibitor to a 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8–5.8 mPa.s) aqueous solution as a continuous phase (W phase) to a final concentration of 2.0 (w / v)%.

[0184] After connecting the continuous phase to an emulsifying device equipped with a 40 μm diameter porous membrane, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing biodegradable polymer microdroplets containing kagrillintide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm. The temperature of the preparation vessel was maintained at 25°C, and after the injection of the dispersed phase, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the temperature was filtered to remove the residual polyvinyl alcohol with triple-distilled water, and freeze-dried.

[0185]

[0186] Example No. Polymer type Polymer amount (g) Kagrilintide amount (g) DCM amount (g) Glacial acetic acid amount (g) Early release inhibitor and concentration 3-1RG503H0.4250.0755.21.4252(w / v)% Na2HPO4 3-2RG653H0.4250.0755.21.4252(w / v)% Na2HPO4 3-3RG753H0.4250.0755.21.4252(w / v)% Na2HPO4 3-4R203H0.4250.0755.21.4252(w / v)% Na2HPO4

[0187] Examples 4-1 to 4-3 (W / O / W method): Preparation of biodegradable polymer microspheres containing kagrilintide and an early-release inhibitor (difference in drug target loading%)

[0188] As a drug, Kagrilintide (manufacturer: Zhejiang Peptides Biotech Co., Ltd, China) was dissolved in 0.665 to 0.770 g of triple-distilled water (DW) to prepare the first aqueous phase (W1 phase). The oil phase (O phase) was prepared by dissolving biodegradable polymers RG653H and RG753H (manufacturer: Evonik, Germany) in a 1:1 weight ratio in 5.2 g of dichloromethane (DCM). The aqueous phase was dispersed in the oil phase using a homogenizer to prepare the first W1 / O emulsion (dispersed phase).

[0189] Na2HPO4 (Disodium phosphate) was added as an initial release inhibitor to a 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8–5.8 mPa.s) aqueous solution as a continuous phase (W2 phase) to a final concentration of 2.0 (w / v)%.

[0190] After connecting the continuous phase to an emulsifying device equipped with a 40 μm diameter porous membrane, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. A secondary W1 / O / W2 emulsion containing biodegradable polymer microdroplets containing kagrillintide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0191] The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase was injected, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the solution was filtered, and the residual polyvinyl alcohol was removed with triple-distilled water, followed by freeze-drying.

[0192]

[0193] Example No. Polymer type Polymer amount (g) Kagrilintide amount (g) DCM amount (g) DW:DCM Weight ratio Initial release inhibitor and concentration 4-1RG653H:RG753H=1:10.4000.1007.61:9.92(w / v)% Na2HPO4 4-2RG653H:RG753H=1:10.3750.1257.11:10.12(w / v)% Na2HPO4 4-3RG653H:RG753H=1:10.3500.1506.71:10.02(w / v)% Na2HPO4

[0194] Examples 5-1 to 5-6 (O / W formulation): Preparation of biodegradable polymer microspheres containing kagrilintide, semaglutide, and an early-release inhibitor (differences in the polymer type of co-encapsulation microspheres, differences in drug target loading %)

[0195] Kagrilintide (manufactured by Zhejiang Peptides Biotech Co., Ltd, China) and semaglutide (manufactured by Chengdu Shengnuo Biopharm Co., Ltd Shengnuo Biopharm Co., Ltd, China) were used as drugs, and the biodegradable polymer was used in a 1:1 weight ratio of RG653H and RG753H (manufactured by Evonik, Germany), or RG63H or RG753H (manufactured by Evonik, Germany) was weighed so that the batch size was 0.5 g. Dichloromethane (DCM) was used as a solvent to prepare the dispersed phase, and glacial acetic acid was used as a cosolvent to homogeneously dissolve it.

[0196] Na2HPO4 (Disodium phosphate) was added as an initial release inhibitor to a 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8-5.8 mPa.s) aqueous solution as a continuous phase (W phase) to a final concentration of 2.0-2.5 (w / v)%.

[0197] After connecting the continuous phase to an emulsifying device equipped with a 40 μm diameter porous membrane, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing biodegradable polymer microdroplets containing kagrillintide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0198] The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase was injected, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the solution was filtered, and the residual polyvinyl alcohol was removed with triple-distilled water, followed by freeze-drying.

[0199]

[0200] Example No. T / L % Polymer type Polymer amount (g) Kagrilintide amount (g) Semaglutide amount (g) DCM amount (g) Glacial acetic acid amount (g) Early release inhibitor and concentration 5-110 / 10RG653H:RG753H=1:10.4000.0500.0504.92.1052(w / v)% Na2HPO45-210 / 10RG653H0.4000.0500.0504.92.4112(w / v)% Na2HPO45-310 / 10RG753H0.4000.0500.0504.92.4112(w / v)% Na2HPO45-412.5 / 12.5RG653H0.3750.06250.06257.12.8002(w / v)% Na2HPO45-512.5 / 12.5RG753H0.3750.06250.06257.12.8002(w / v)% Na2HPO45-615 / 15RG753H0.3500.0750.0756.73.0242.5(w / v)% Na2HPO4

[0201] * T / L:Target Loading (cagrilintide / semaglutide)

[0202] Examples 6-1 to 6-6 (microsphere blending): Preparation of blending of caprylintide microspheres and semaglutide microspheres

[0203] Semaglutide microspheres (A) and caprylintide microspheres (B) were mixed in a mixing ratio (weight ratio) A:B of 2:1 to 1:2, respectively, and the content of each drug in the microsphere blend was confirmed.

[0204] Example Semaglutide microspheres (A) Semaglutide content Kagrilintide microspheres (B) Kagrilintide content Mixing ratio (A:B) 6-1 Manufacturing example 116.0% Example 1-2 17.35% 1:16-2 Manufacturing example 116.0% Example 1-2 17.35% 2:16-3 Manufacturing example 116.0% Example 1-2 17.35% 1:26-4 Manufacturing example 213.3% Example 4-118.3% 1:16-5 Manufacturing example 213.3% Example 4-118.3% 2:16-6 Manufacturing example 213.3% Example 4-118.3% 1:2

[0205] The above semaglutide microspheres were manufactured through Manufacturing Examples 1 and 2 below.

[0206] Manufacturing Example 1 (O / W Manufacturing Method): Manufacturing of Semaglutide-Containing Polymer Microparticles

[0207] Semaglutide freebase (0.450 g) as a drug (manufacturer: Chengdu Shengnuo Biopharm Co., Ltd, China) and biodegradable polymer (RG653H:RG753H = 1:1) (1.60 g) were weighed to obtain a batch size of 2.0 g. Dichloromethane was used as a solvent for preparing the dispersed phase, and glacial acetic acid was used as a cosolvent to prepare an oil phase (O phase) in which the drug and polymer were homogeneously dissolved.

[0208] Na2HPO4 (Disodium phosphate) was added as an initial release inhibitor to a 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8-5.8 mPa.s) aqueous solution as a continuous phase (W phase) to a final concentration of 2.0-2.5 (w / v)%.

[0209] After connecting the continuous phase to an emulsifying device equipped with a 40 μm diameter porous membrane, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing microdroplets of a biodegradable polymer including semaglutide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0210] The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase was injected, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the solution was filtered, and the residual polyvinyl alcohol was removed with triple-distilled water, followed by freeze-drying.

[0211]

[0212] Manufacturing Example 2 (O / W method): Manufacturing of polymer microspheres containing semaglutide

[0213] Semaglutide freebase (0.450 g) as a drug (manufacturer: Chengdu Shengnuo Biopharm Co., Ltd, China) and biodegradable polymer (RG653H:RG753H = 1:1) (2.55 g) were weighed to obtain a batch size of 3.0 g. Dichloromethane was used as a solvent for preparing a dispersed phase, and an oil phase (O phase) in which the drug and polymer were homogeneously dissolved was prepared as a dispersed phase.

[0214] Na2HPO4 (Disodium phosphate) was added as an initial release inhibitor to a 0.1 (w / v) % polyvinyl alcohol (viscosity: 4.8-5.8 mPa.s) aqueous solution as a continuous phase (W phase) to a final concentration of 2.0-2.5 (w / v)%.

[0215] After connecting the continuous phase to an emulsifying device equipped with a 40 μm diameter porous membrane, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing microdroplets of a biodegradable polymer including semaglutide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0216] The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase was injected, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the solution was filtered, and the residual polyvinyl alcohol was removed with triple-distilled water, followed by freeze-drying.

[0217]

[0218] Comparative Example 1 (O / W method): Preparation of biodegradable polymer microspheres containing kagrillintide without initial release inhibitor

[0219] Kagrilintide (0.075 g, manufactured by Zhejiang Peptides Biotech Co., Ltd, China) was used as the drug, and RG653H and RG753H (manufactured by Evonik, Germany) were weighed at 0.425 g in a 1:1 weight ratio for a batch size of 0.5 g. Dichloromethane (DCM) was used as the solvent for preparing the dispersed phase, and glacial acetic acid was used as a cosolvent to homogeneously dissolve it. 2,000 ml of a 0.1 (w / v)% polyvinyl alcohol (viscosity: 4.8–5.8 mPa.s) aqueous solution was used as the continuous phase. The continuous phase was connected to an emulsifying device equipped with a 40 μm diameter porous membrane, and the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion containing biodegradable polymer microdroplets containing kagrilintide was prepared, and the suspension was placed in a preparation container and stirred at a speed of 200 to 300 rpm.

[0220] The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase was injected, the temperature of the suspension was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the solution was filtered, and the residual polyvinyl alcohol was removed with triple-distilled water, followed by freeze-drying.

[0221] Comparative Example No. Polymer Type Polymer Amount Used (g) Kagrilintide Amount Used (g) DMC Amount Used (g) Glacial Acetic Acid Amount Used (g) Comparative Example 1 RG653H:RG753H=1:10.4250.0755.21.433

[0222] Comparative Example 2 (W / O / W method): Preparation of biodegradable polymer microspheres containing kagrillintide without initial release inhibitor

[0223] The first phase (W1) was prepared by dissolving 0.075 g of Kagrilintide (manufacturer: Zhejiang Peptides Biotech Co., Ltd, China) as a drug in 0.675 g of distilled water (DW). The oil phase (O) was prepared by mixing RG653H and RG753H (manufacturer: Evonik, Germany) as biodegradable polymers at a weight ratio of 1:1 to make 0.425 g and dissolving the mixture in 3.7 g of dichloromethane (DCM). The first W1 / O emulsion (dispersed phase) was prepared by dispersing the aqueous phase in the organic phase using a homogenizer. The continuous phase (W2) was prepared by using 2,000 mL of a 0.1 (w / v)% polyvinyl alcohol (viscosity: 4.8–5.8 mPa·s) aqueous solution. After connecting the continuous phase to an emulsifying device equipped with a porous membrane having a diameter of 40 μm, the prepared dispersed phase was injected into the porous membrane together with the continuous phase. A secondary W1 / O / W2 emulsion in which biodegradable polymer microdroplets containing kagrillintide were dispersed was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0224] The temperature of the preparation vessel was maintained at 25°C, and after the dispersion phase injection was completed, the suspension temperature was maintained at 40°C for 3 hours to remove the organic solvent. After cooling to 25°C, the solution was filtered to remove the remaining polyvinyl alcohol with triple-distilled water, and freeze-dried.

[0225] Comparative Example No. Polymer Type Polymer Amount Used (g) Kagrilintide Amount Used (g) DCM Amount Used (g) DW:DCM Weight Ratio Comparative Example 2 RG653H:RG753H=1:10.4250.0753.71:5.48

[0226] Experimental Example 1. Measurement of the amount of caprylintide encapsulated in microparticles.

[0227] In order to measure the drug content of the microspheres manufactured in the above example, 10 mg of microspheres were completely dissolved in 4.0 mL of acetonitrile, and then extracted by adding 6.0 mL of 0.1% (w / w) trifluoroacetic acid aqueous solution. The solution filtered using a 0.45 μm filter was used as a test solution. 20 μL of the test solution was injected into HPLC and measured at a detection wavelength of 214 nm. The column used in this experimental example was ZORBAX 300SB-C18, 5 μm, 4.6 x 150 mm, and the mobile phase was a mixture of acetonitrile containing 0.1% (w / w) trifluoroacetic acid and 0.1% (w / w) trifluoroacetic acid aqueous solution in a ratio of 40:60 (v / v).

[0228] Kagrilintide loading rate (%) Kagrilintide content (wt%) Semaglutide loading rate (%) Semaglutide content (wt%) Example 1-192.10 13.81 N.AN.A Example 1-286.70 17.35 N.AN.A Example 1-374.20 18.56 N.AN.A Example 1-484.85 25.46 N.AN.A Example 1-589.21 31.22 N.AN.A Example 2-1101.97 15.30 N.AN.A Example 2-2103.40 15.51 N.AN.A Example 2-3103.04 15.46 N.AN.A Example 2-466.589.99N.AN.A Example 3-190.9013.64N.AN.A Example 3-295.0014.25N.AN.A Example 3-399.4914.92N.AN.A Example 3-498.4014.76N.AN.A Example 4-191.5518.31N.AN.A Example 4-294.1823.55N.AN.A Example 4-391.1427.34N.AN.A Example 5-192.809.2887.518.75 Example 5-285.228.5285.758.58 Example 5-386.828.6887.028.70Example 5-481.3210.1783.9810.50Example 5-588.4311.0591.5911.45Example 5-677.2811.5984.2612.64Comparative Example 1106.3015.94N.AN.AComparative Example 299.5014.92N.AN.A

[0229] As shown in Table 9,

[0230] Looking at the results of Examples 1-1 to 1-5 with different drug T / L contents in the O / W formulation, it can be confirmed that the encapsulation rate tends to be maintained at 70% or more up to T / L 35% (Example 1-5).

[0231] Looking at the results of Examples 4-1 to 4-3 with different drug T / L contents in the W / O / W formulation, it can be confirmed that the encapsulation rate is maintained at 90% or more up to T / L 30% (Example 4-3).

[0232] Looking at the results of Examples 2-1 to 2-4 using different types of early-release inhibitors in the O / W method, in the case of Example 2-4 using (NH4)2HPO4 (Diammonium phosphate), the encapsulation rate drops sharply to less than 70%, but the remaining three types of early-release inhibitors tend not to affect the encapsulation rate.

[0233] Looking at the results of Examples 3-1 to 3-4 in which the type of polymer was changed in the O / W method, the type of polymer did not have much effect on the encapsulation rate.

[0234] Looking at the results of Co-encapsulation microsphere examples 5-1 to 5-3 using different polymer types in the O / W method, the polymer type did not have a significant effect on the encapsulation rate.

[0235] Looking at the results of Co-encapsulation microsphere examples 5-3, 5-5, and 5-6 with different drug T / L contents in the O / W method, it can be confirmed that as the drug T / L content increases, the encapsulation rate decreases, but it is still maintained at 70% or more.

[0236]

[0237] Experimental Example 2. In vitro measurement of initial drug release from caprylintide microspheres

[0238] To confirm the initial drug release of the microspheres manufactured in the above examples and comparative examples, the following experiment was conducted. 10 mg of microspheres were placed in an HDPE wide-mouth bottle, filled with 50 mL of a release test solution, and stored in a 37°C incubator. After 24 hours, 1 mL of the test solution was taken, centrifuged, and the supernatant obtained was analyzed for the content and release rate of kagrilintide and semaglutide using HPLC under the same analysis conditions as in Experimental Example 1. The release test solution used in this measurement was an aqueous solution containing 0.1% polysorbate 20 at pH 7.4.

[0239] In vitro initial release rate (%, 1-day) Kagrilintide Semaglutide Example 1-10.96NA Example 1-21.54NA Example 1-34.73NA Example 1-43.99NA Example 1-55.81NA Example 1-66.92NA Example 1-77.65NA Example 2-10.30NA Example 2-20.25NA Example 2-30.24NA Example 2-40.51NA Example 3-10.18NA Example 3-20.36NA Example 3-30.42NA Example 3-40.85NA Example 4-13.40NA Example 4-23.44NA Example 5-12.682.82 Example 5-21.991.48 Example 5-32.331.96 Example 5-45.025.12 Example 5-54.995.23 Example 5-67.097.41

[0240] As shown in Table 10, it was confirmed that the initial release of the drug was significantly reduced in microspheres containing only kagrillintide and in co-encapsulated microspheres containing kagrillintide and semaglutide when an initial release inhibitor was included in the continuous phase during the manufacture of microspheres.

[0241]

[0242] Experimental Example 3. Morphological Analysis Using an Electron Microscope

[0243] The morphological characteristics of the microspheres according to the present invention were analyzed using an electron microscope. The experimental procedure is as follows. 5 mg of the microspheres prepared in the examples and comparative examples were placed on an aluminum stub with a carbon tape attached and coated with platinum using an ION-COATER (COXEM, Korea). The aluminum stub was mounted on a scanning electron microscope (COXEM EM-30, Korea), and the morphological characteristics of the microspheres were observed at an acceleration voltage of 10 to kV, and the results are shown in Figures 1 to 6.

[0244]

[0245] Figure 1 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 1-1 to 1-7.

[0246] Figure 2 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 2-1 to 2-4.

[0247] Figure 3 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 3-1 to 3-4.

[0248] Figure 4 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Examples 4-1 to 4-3.

[0249] Figure 5 is a scanning electron microscope photograph confirming the morphological characteristics of co-encapsulation microspheres manufactured in Examples 5-1 and 5-6.

[0250] Figure 6 is a scanning electron microscope photograph confirming the morphological characteristics of microspheres manufactured in Comparative Examples 1 and 2.

[0251]

[0252] As shown in Figures 1 to 6, it was confirmed that all microspheres manufactured by the O / W method had a uniform and smooth surface, and it was confirmed that the microspheres manufactured by the W / O / W method were uniform, but some surfaces were not smooth, but were of good quality.

[0253]

[0254] Experimental Example 4. In-vivo pharmacokinetic evaluation using rats

[0255] In order to evaluate the drug release profile of microspheres according to an embodiment of the present invention, the concentration of kagrilintide in the blood was measured after administration to rats.

[0256] Microspheres were weighed to be 1.2 mg / head (4.0 mg / kg) or 3.6 mg / head (12.0 mg / kg) of kagrillintide, dispersed in 0.5 mL of suspension, and subcutaneously injected into SD rats (Sprague-Dawley rats, 300 g). 0.5 mL of blood was collected at pre-scheduled times, and the blood concentration of kagrillintide was measured using HPLC. Here, in the case of co-encapsulation microspheres (Examples 5-1 and 5-6), the blood concentration of kagrillintide, excluding semaglutide, was analyzed.

[0257]

[0258] Experiment no. G4G5G6G7G18Example no. 1-11-21-35-15-6T / L %(cagrilintide / semaglutide) 15202510 / 1015 / 15Dose(mg / head) 3.63.63.63.61.2AUCnorm(ng*day / mL) / (mg / head) 208.6651.21796.44898.85486.7Cmax(ng / ml) 41.7173.4690.11512.8407.23

[0259] As shown in Table 11, it was confirmed that the bioavailability of the co-encapsulated microspheres of kagrillintide and semaglutide (Examples 5-1 and 5-6) was significantly improved compared to the microspheres of kagrillintide alone (Examples 1-1 to 1-3).

[0260]

[0261] Experimental Example 5. Single-dose subcutaneous pharmacokinetic study using Sprague-Dawley rats

[0262] In order to evaluate the potential of the microspheres of the present invention as a sustained-release therapeutic agent, the concentration of caprylintide in rat blood was measured by the following method.

[0263] Specifically, the dose of kagrillintide was 1.2 mg / head (API raw material solution, Example 5-6) or 3.6 mg / head (Comparative Examples 1 to 2, Examples 1-1 to 1-3, Example 5-1), and microspheres were measured, dispersed in 0.5 mL of suspension, and then subcutaneously injected into SD rats. 0.25-0.5 mL of blood was collected at pre-scheduled times, and the concentration of kagrillintide in the blood was measured using HPLC.

[0264]

[0265]

[0266]

[0267] As shown in Table 12,

[0268] The microspheres of Comparative Examples 1 and 2, which did not use an initial release inhibitor, showed an initial burst phenomenon with a cumulative blood concentration exceeding 10% on the first day, whereas the microspheres of Examples 1-1 to 1-3, Example 5-1, and Example 5-6 all showed a cumulative blood concentration of 10% or less on the first day, and showed an excellent sustained release profile with continuous release for a short period of 56 days and a long period of 84 days.

[0269]

[0270] Experimental Example 6. Measurement of residual early-release inhibitor content in microparticles.

[0271] In order to measure the content of early release inhibitors in the microspheres manufactured in the above example, the residual amounts of sodium (Na) and phosphorus (P) in the microspheres were measured.

[0272] Specifically, 300 mg of microspheres were mixed with 6 mL of a nitric acid solution mixed 1:1 with ultrapure water and 3 mL of hydrogen peroxide, heated to above 100°C, and acid was added until the gas generated during the dissolution process changed from yellow to white. The sample obtained through this was weighed, dissolved in ultrapure water, and injected into an inductively coupled plasma optical emission spectrometer (ICP-OES) (Thermo Scientific Co., iCAP 6300 Duo, UK) and measured at a detection wavelength of 598.5 nm.

[0273] Example 3-4 Example 1-1 Example 5-6 Example 2-1 Early release inhibitor 2 (w / v) % Na2HPO4 2 (w / v) % Na2HPO4 2.5 (w / v) % Na2HPO4 2 (w / v) % K2HPO4 Polymer R20 3 HRG 653 H:RG 753 HRG 753 HRG 653 H:RG 753 H Residual sodium (mg / kg) 399 105 171 - Residual phosphorus (mg / kg) 813 35441

[0274] As shown in Table 13, the content of residual sodium and phosphorus according to the initial release inhibitor included in the continuous phase differs depending on the microparticle manufacturing conditions, and it was confirmed that the content of the initial release inhibitor remaining in the microparticles calculated from this was 50 ppm to 500 ppm based on sodium (Na) and 10 to 100 ppm based on phosphorus (P).

Claims

1. As an active ingredient, (i) kagrillintide or a pharmaceutically acceptable salt thereof, or (ii) kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof; Early-release inhibitors; and Comprising western-type microspheres containing biodegradable polymers, The above (i) Kagrilintide or a pharmaceutically acceptable salt thereof is contained in an amount of 8 wt% or more as Kagrilintide based on the total weight of the microspheres, The above (ii) Kagrilintide or a pharmaceutically acceptable salt thereof is contained in an amount of 4 wt% or more as Kagrilintide based on the total weight of the microspheres, A pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta-cell dysfunction, obesity, or non-alcoholic steatohepatitis, or Alzheimer's disease, wherein the early-release inhibitor is included in an amount of 10 to 2,000 ppm.

2. Sustained-release microspheres comprising kagrillintide or a pharmaceutically acceptable salt thereof, an early-release inhibitor and a biodegradable polymer as active ingredients; and A combination of sustained-release microspheres comprising semaglutide or a pharmaceutically acceptable salt thereof as an active ingredient, an early-release inhibitor; and a biodegradable polymer, The above-mentioned kagrillintide or a pharmaceutically acceptable salt thereof is contained in an amount of 8 wt% or more as kagrillintide based on the total weight of the microspheres, A pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta-cell dysfunction, obesity, or non-alcoholic steatohepatitis, or Alzheimer's disease, wherein the early-release inhibitor is included in an amount of 10 to 2,000 ppm.

3. A pharmaceutical composition according to claim 1 or 2, wherein the release rate of kagrilintide or a pharmaceutically acceptable salt thereof in the microspheres is less than 20% within 24 hours when administered in vivo.

4. A pharmaceutical composition according to claim 1 or 2, wherein the release rate of kagrilintide or a pharmaceutically acceptable salt thereof in the microspheres is less than 15% within 24 hours when administered in vivo.

5. A pharmaceutical composition according to claim 1 or 2, wherein the release rate of kagrilintide or a pharmaceutically acceptable salt thereof in the microspheres is less than 10% within 24 hours when administered in vivo.

6. In the first or second paragraph, the biodegradable polymer Polylactide (PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), a copolymer of lactide and glycol, polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkylcarbonate, polytrimethylenecarbonate (PTMC), polylactide-co-trimethylenecarbonate (PLTMC), A polymer selected from the group consisting of polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo polyaminoacid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphoester, polysaccharide, and poly(butylene succinate lactide) (PBSLA); a copolymer or simple mixture of two or more thereof; a copolymer of the above polymer and polyethylenglycol (PEG);A pharmaceutical composition comprising at least one selected from the group consisting of a polymer-sugar complex in which the polymer or copolymer is combined with a sugar.

7. A pharmaceutical composition according to claim 1 or 2, wherein the early release inhibitor is at least one substance selected from among phosphate salts, hydroxide salts, phosphide salts, phosphite salts, carbonate salts, chromate salts, dichromate salts, oxides, oxalate salts, silicate salts, sulfate salts, sulfide salts, sulfite salts, tartrate salts, tetraborate salts, thiosulfate salts, arsenate salts, arsenite salts, citrate salts, ferricyanide salts, and nitride salts of alkali metals, alkaline earth metals, or ammonium.

8. A pharmaceutical composition according to claim 1 or 2, wherein the average particle size of the microspheres containing the kagrilintide or a pharmaceutically acceptable salt thereof and the early release inhibitor is 5 μm to 100 μm.

9. A pharmaceutical composition according to claim 1 or 2, wherein the weight of the microspheres containing the kagrilintide or a pharmaceutically acceptable salt thereof and an early-release inhibitor is 20 to 1000 mg.

10. A pharmaceutical composition according to claim 1 or 2, wherein the intrinsic viscosity of the biodegradable polymer is 0.16 dL / g to 1.7 dL / g.

11. A pharmaceutical composition according to claim 1 or 2, wherein the microspheres further comprise at least one release-controlling agent selected from the group consisting of butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, nonadecylic acid, behenic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, napadicylic acid, naphthalene sulfonic acid, and pamoic acid.

12. In paragraph 1, A pharmaceutical composition, wherein the semaglutide or a pharmaceutically acceptable salt thereof of the above (ii) is contained in an amount of 4 wt% or more as semaglutide based on the total weight of the microspheres.

13. In paragraph 2, A pharmaceutical composition wherein the semaglutide or a pharmaceutically acceptable salt thereof is contained in an amount of 8 wt% or more as semaglutide based on the total weight of the microspheres. 14.(a) A step of preparing an oil phase (O phase) in which (i) kagrillintide or a pharmaceutically acceptable salt thereof, or (ii) kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof, and one or more biodegradable polymers are dissolved in an organic solvent as an active ingredient; (b) a step of preparing a continuous phase (W phase) by adding an early release inhibitor to an aqueous solution containing a surfactant, and adding the dispersed phase of step (a) to prepare a dispersed phase in an emulsion state; (c) a step of extracting and evaporating an organic solvent from the dispersed phase of the emulsion state prepared in step (b) into a continuous phase (W phase) to form microspheres; and (d) a method for preparing microspheres with an O / W emulsion comprising an active ingredient, an early release inhibitor and a biodegradable polymer, comprising a step of recovering the microspheres. 15.(a') A step of preparing a first water phase (W1 phase) by dissolving (i) kagrillintide or a pharmaceutically acceptable salt thereof or (ii) kagrillintide or a pharmaceutically acceptable salt thereof and semaglutide or a pharmaceutically acceptable salt thereof as an active ingredient in an aqueous solution, and preparing an oil phase (O phase) by dissolving one or more biodegradable polymers in an organic solvent, and preparing a first W1 / O emulsion by mixing the water phase (W1 phase) and the oil phase (O phase) as a dispersion phase; (b') A step of preparing a continuous phase (W2 phase) by adding an early release inhibitor to an aqueous solution containing a surfactant, and adding the dispersed phase of step (a') to prepare a dispersed phase in a secondary W1 / O / W2 emulsion state; (c') a step of forming microspheres by extracting and / or evaporating the organic solvent from the dispersed phase of the second W1 / O / W2 emulsion state of the above step (b') into a continuous phase (W2 phase); and (d') A method for preparing microspheres using a W / O / W emulsion comprising an active ingredient, an early release inhibitor and a biodegradable polymer, comprising a step of recovering microspheres.

16. In any one of claims 14 to 15, the biodegradable polymer is polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA), polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkylcarbonate, polytrimethylenecarbonate (PTMC), polylactide-co-trimethylenecarbonate (PTMC), A polymer selected from the group consisting of polyhydroxybutyric acid (PLTMC), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo polyaminoacid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphoester, polysaccharide and poly(butylene succinate lactide) (PBSLA); a copolymer or simple mixture of two or more thereof;A manufacturing method comprising at least one selected from the group consisting of a copolymer of the above polymer and polyethylene glycol (polyethylenglycol, PEG); and a polymer-sugar complex in which the above polymer or copolymer is combined with sugar.

17. A manufacturing method according to any one of claims 14 to 15, wherein the dispersed phase of step (a) or (a') further comprises at least one release controlling agent selected from the group consisting of butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, nonadecylic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, hydroxynaphthoic acid, napadicylic acid, and pamoic acid.

18. A manufacturing method according to any one of claims 14 to 15, wherein the organic solvent of step (a) or (a') is at least one organic solvent 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.

19. A manufacturing method according to any one of claims 14 to 15, wherein the surfactant of step (b) or (b') is polyvinyl alcohol.

20. A manufacturing method according to any one of claims 14 to 15, wherein the continuous phase of step (b) or (b') further includes at least one selected from the group consisting of methyl alcohol, ethyl alcohol, propyl alcohol, and ethyl acetate.

21. A manufacturing method according to any one of claims 14 to 15, wherein the continuous phase of step (b) is used by adding an early release inhibitor to an aqueous solution containing a surfactant so that the final concentration is 0.1 to 5.0 (w / v)%.

22. A manufacturing method according to any one of claims 14 to 15, wherein the early release inhibitor is at least one selected from the group consisting of phosphate salts of two or more alkali metals, bicarbonate salts of one or more alkali metals, and carbonate salts of one or more alkali metals.

23. A manufacturing method according to any one of claims 14 to 15, wherein the pH of the continuous phase of step (b) or (b') is 7 or higher.

24. A manufacturing method according to any one of claims 14 to 15, wherein the release rate of kagrillintide or a pharmaceutically acceptable salt thereof is less than 15% within 24 hours when the manufactured sustained-release microspheres are administered into a living body.

25. A manufacturing method according to any one of claims 14 to 15, wherein the release rate of kagrillintide or a pharmaceutically acceptable salt thereof is less than 10% within 24 hours when the manufactured sustained-release microspheres are administered into a living body.

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