Sustained-release microsphere preparation containing semeglutide or pharmaceutically acceptable salt thereof and preparation method of sustained-release microsphere preparation

Abstract

The present invention relates to a pharmaceutical composition containing sustained release microspheres composed of semeglutide or a pharmacologically acceptable salt thereof, a bioavailability enhancer and a biodegradable polymer, which is designed to avoid rapid initial drug release (initial burst release), contains a high concentration of drug with respect to particle size, has high bioavailability, and can be used in the field of pharmaceutical preparations. And when the composition is applied to a human body, discomfort and inflammatory response of a patient are reduced as much as possible, so that the composition can be used for preventing or treating degenerative nervous system diseases such as diabetes, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis or Alzheimer's disease and Parkinson's disease and can be used for protecting beta cell functions.

Description

Technical Field

[0001] The present disclosure relates to a sustained-release microsphere containing semaglutide or a pharmaceutically acceptable salt thereof with enhanced bioavailability, a pharmaceutical composition comprising the sustained-release microsphere containing a high content of semaglutide, and a method for preparing the sustained-release microsphere. Background Art

[0002] Glucagon-like peptide-1 (GLP-1) is a peptide derived from preproglucagon. Preproglucagon is a 158 amino acid precursor polypeptide that is differentially processed in tissues to form many different proglucagon-derived peptides, including glucagon, glucagon-like peptide-1 (GLP-1), glucagon-like peptide-2 (GLP-2) and oxyntomodulin (OXM). These molecules are involved in a variety of physiological functions, including the regulation of glucose homeostasis, insulin secretion, gastric emptying and intestinal growth and food intake. GLP-1 is produced as a 37 amino acid peptide that corresponds to amino acids 72 to 108 of proglucagon (preproglucagon 92 to 128). GLP-1 (7-36) amide or GLP-1 (7-37) acid is the biologically active form of GLP-1, which exhibits substantially equivalent activity to the GLP-1 receptor. GLP-1 and GLP-1 analogs, which are GLP-1 receptor agonists, have been shown to provide effective glycemic control, for example for the treatment of patients with type 2 diabetes, and additionally provide weight loss effects, preserve beta cell function, and reduce hypertension, hypoglycemia, and / or hyperlipidemia. Bydurepn Bydureon (exenatide), (Semaglutide), (liraglutide), (lixisenatide), (albiglutide) and Some GLP-1 analogs, including Glucosamine (dulaglutide), are either commercially available or under development.

[0003] GLP-1 agonists (such as semaglutide) are peptides that are mainly administered by injection due to the presence of several barriers (such as enzymatic degradation in the gastrointestinal tract and intestinal mucosa, insufficient intestinal mucosal absorption, and first-pass metabolism in the liver). Recently, oral administration agents have also been commercialized, but their bioavailability is much lower than that of injection preparations and significantly higher doses are required. In addition, dosage forms containing semaglutide are formulated to be directly administered by the patient himself (self-administration) for sustainable management of obesity or diabetes, so it is very important to regulate pain, inflammatory reactions, etc. that may occur at the application site.

[0004] On the other hand, it is known to encapsulate semaglutide in microparticles made of biodegradable polymers for long-term release. In this case, due to the low bioavailability of semaglutide encapsulated in the microspheres or the low drug content in the microspheres, a large number of microspheres must be administered to show a long-term effective pharmacological effect. However, a large number of microspheres in the body are difficult to administer subcutaneously, making it difficult for patients to directly administer (self-administer) drugs, which can cause severe pain and extremely high inflammatory reactions at the site of administration. Summary of the invention

[0005] Technical issues

[0006] The present disclosure aims to solve the above-mentioned problems, and the purpose of the present disclosure is to provide: a pharmaceutical composition comprising sustained-release microspheres composed of semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a biodegradable polymer, wherein the semaglutide has high bioavailability and has long-term stable drug release performance; and a pharmaceutical composition comprising sustained-release microspheres composed of semaglutide or a pharmaceutically acceptable salt thereof and a biodegradable polymer, which exhibits stable drug release performance with high drug content and low initial release; and a method for preparing sustained-release microspheres.

[0007] Technical Solution

[0008] The present disclosure is described in detail below.

[0009] In the present disclosure, the term "at least one" refers to a number corresponding to one or more. In the present disclosure, when the quantity of any component 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.

[0010] To achieve the above-mentioned object, in one aspect of the present disclosure, a pharmaceutical composition is provided, which comprises semaglutide sustained-release microspheres containing a biodegradable polymer, wherein the content of semaglutide is at least 8 wt % based on the total weight of the microspheres as semaglutide, and wherein the content of the bioavailability enhancer is 2.5 wt % to 250 wt % based on the weight of semaglutide.

[0011] In another aspect of the present disclosure, a pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, protection of β-cell function, obesity, non-alcoholic fatty hepatitis or neurodegenerative diseases is provided, the pharmaceutical composition comprising sustained-release microspheres composed of semaglutide or a pharmaceutically acceptable salt thereof, an initial release inhibitor and a biodegradable polymer, wherein the content of semaglutide or a pharmaceutically acceptable salt thereof is 8 wt % or more based on the total weight of the microspheres as semaglutide, and the content of the initial release inhibitor is 5 ppm to 2,000 ppm.

[0012] On the other hand, the biodegradable polymer may be at least one selected from the group consisting of polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA) which is a copolymer of lactide and glycolide, polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co-trimethylene carbonate (PLTMC). , polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoesters, polyanhydrides, polyanhydride-co-imides, polypropylene fumarate, pseudo-polyamino acids, polyalkyl cyanoacrylates, polyphosphazenes, polyphosphates, block copolymers of poly(butylene succinate / lactide) (PBSLA); copolymers or simple mixtures of two or more thereof; copolymers of a polymer and polyethylene glycol (PEG); and at least one selected from a polymer-sugar complex in which a polymer or a copolymer is conjugated to a sugar.

[0013] In another aspect, the bioavailability enhancer can be at least one selected from the group consisting of sodium caprate, disodium hydrogen phosphate, choline, meglumine, basic aluminum carbonate, sodium dihydroxyaluminum carbonate, ammonium phosphate, histidine, HEPES, HEPPS, spermine, spermidine, putrescine, methylene blue, proline, sugar, glycerol, surfactant, arginine, glycine, guanidine hydrochloride, urea, sodium chloride, potassium chloride, triethylamine, ethanolamine, triethanolamine, ethylenediamine, poloxamer, benzathine penicillin, e), procaine, lidocaine, bupivacaine, ropivacaine, oxytetracycline, sunitinib, rhizolutin, benzofuran, magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium trisilicate, zinc carbonate, zinc hydroxide, zinc phosphate, aluminum hydroxide, aluminum phosphate, dihydroxyaminoacetic acid aluminum, calcium phosphate, calcium hydroxide, magnesium aluminum hydroxide and benzofuran derivatives.

[0014] In another aspect, the pharmaceutically acceptable salt of semaglutide is the sodium salt, acetate, benzoate, hydroxynaphthoate, naphalene disulfonate or pamoate of semaglutide.

[0015] In another aspect, the intrinsic viscosity of the poly(lactide-co-glycolide), polyglycolide, or polylactide in the biodegradable polymer is from 0.16 dL / g to 1.7 dL / g.

[0016] In another aspect, the microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer may have an average particle size of 5 μm to 100 μm.

[0017] In another aspect, the microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer may have a span value of 1.5 or less.

[0018] In another aspect, the weight of the microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer can be 20 mg to 1,000 mg, 20 mg to 800 mg, 20 mg to 600 mg, 20 mg to 400 mg, 20 mg to 200 mg, 20 mg to 100 mg, 30 mg to 1,000 mg, 30 mg to 800 mg, 30 mg to 600 mg, 30 mg to 400 mg, 30 or 50 mg to 100 mg. In some embodiments, the present invention relates to an oral dosage form of at least 200 mg to 400 mg of the present invention. In some embodiments, the present invention relates to an oral dosage form of at least 200 mg to 400 mg of the present invention.

[0019] On the other hand, one or more release controlling agents can also be included, and the one or more release controlling agents are selected from the group consisting of butyric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, behexadecanoic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, naphthadicylic acid, naphthalenesulfonic acid and pamoic acid.

[0020] In another aspect, the microspheres can contain 1 mg / kg to 100 mg / kg of Na.

[0021] In another aspect, the microspheres can contain 10 mg / kg to 500 mg / kg of P.

[0022] In another aspect of the present disclosure, a method for preparing sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer is provided.

[0023] In another aspect, the method of preparation can employ a continuous phase containing an initial release inhibitory agent.

[0024] On the other hand, when preparing microspheres containing semaglutide, the initial release inhibitor may be a salt dissolved in the continuous phase to maintain the pH of the continuous phase at 7.0 or higher.

[0025] On the other hand, the initial release inhibitor can be at least one substance that forms one or more anions, the substance being selected from alkali metal, alkaline earth metal or ammonium phosphate salt, phosphide salt, carbonate, chromate, dichromate, oxide, oxalate, silicate, sulfate, sulfide salt, sulfite, tartrate, tetraborate, thiosulfate, arsenate, arsenite, citrate, ferrocyanide salt and nitride salt. On the other hand, the initial release inhibitor can be at least one selected from the group consisting of disodium hydrogen phosphate, dipotassium hydrogen phosphate and diammonium hydrogen phosphate.

[0026] In another aspect, provided is a pharmaceutical composition comprising sustained-release microspheres and a method for preparing the same, wherein the sustained-release microspheres contain a biodegradable polymer, wherein the content of semaglutide is 8 wt % or more based on the total weight of the microspheres as semaglutide, and the initial release of the drug is less than 10%.

[0027] Beneficial Effects

[0028] The sustained-release pharmaceutical composition comprising semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the preparation examples disclosed herein can contain a high content of the drug relative to the particle size and has a high bioavailability when administered into the body, thereby reducing the dosage while showing a lasting effect. Therefore, it can minimize the pain and inflammatory response that may occur in patients when administered. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a graph showing changes in blood drug concentration over time after the microspheres containing semaglutide prepared in Preparation Example 12 were administered to rats.

[0030] Figure 2 is a graph showing the initial release of the semaglutide-containing microspheres prepared in Preparation Example 5, Preparation Example 6, Preparation Example 10, and Preparation Example 15.

[0031] Figure 3ais a scanning electron microscope (SEM) photograph showing the cross-sectional surface of microspheres prepared using only 0.1 (w / v)% of PVA in the continuous phase.

[0032] Figure 3b is a scanning electron micrograph showing the cross-sectional surface of microspheres prepared using 0.1 (w / v)% PVA and 2 (w / v)% Na2HPO4 in the continuous phase. DETAILED DESCRIPTION

[0033] Hereinafter, the present disclosure will be described in detail.

[0034] The present disclosure comprises semaglutide or a pharmaceutically acceptable salt thereof as an active ingredient.

[0035] Semaglutide is a GLP-1 receptor agonist 6.26 -{18-[N-(17-carboxy-heptadecanoyl)-L-γ-glutamyl]-10-oxo-3,6,12,15-tetraoxa-9,18-diazaoctadecanoyl}-[8-(2-amino-2-propionic acid), 34-L-arginine] human glucagon-like peptide 1 (7-37), may also be referred to as N-ε26-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8,Arg34]GLP-1-(7-37). The structure of semaglutide is represented by the following chemical formula 1.

[0036] [Chemical formula 1]

[0037]

[0038] Such semaglutide can be prepared as described in Preparation Example 4 of International Patent Publication No. WO2006 / 097537, and commercially available semaglutide can also be used.

[0039] Semaglutide can exist in the form of a salt, in particular a pharmaceutically acceptable salt. As a salt, salts commonly used in the art can be used without restriction. The term "pharmaceutically acceptable salt" as used herein refers to any organic or inorganic addition salt of a compound, the concentration of which shows a relatively non-toxic and harmless effective effect on the patient and the side effects of the salt do not reduce the beneficial effects of the above-mentioned compound. Specific examples include, but are not limited to, sodium salt, acetate, benzoate, hydroxynaphthoate, naphthalene disulfonate or pamoate of Semaglutide.

[0040] Semaglutide or a pharmaceutically acceptable salt thereof as an active ingredient of the present disclosure may be in various forms, for example, amorphous or crystalline.

[0041] In the present disclosure, a "bioavailability enhancer" is a substance that is encapsulated in microparticles together with the active ingredient semaglutide or a pharmaceutically acceptable salt thereof to enhance bioavailability. Specifically, when it is contained in sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and the microspheres are administered into the body, the bioavailability enhancer refers to a substance that increases the area under the blood concentration-time curve (AUC) of semaglutide compared to microspheres that do not contain the "bioavailability enhancer". More specifically, when the sustained-release microspheres are administered to rats, the bioavailability enhancer refers to the area under the blood concentration-time curve (AUC) of semaglutide when 1 mg / kg is administered. 总 / (mg / kg)) shows 600ng*day / mL or more, preferably 1,000ng*day / mL or more, more preferably 1,300ng*day / mL or more.

[0042] In a specific aspect, the bioavailability enhancer can be at least one selected from the group consisting of sodium decanoate, disodium hydrogen phosphate, choline, meglumine, basic aluminum carbonate, sodium dihydroxyaluminum carbonate, ammonium phosphate, histidine, HEPES (hydroxyethylpiperazineethanesulfonic acid) HEPPS (4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid), spermine, spermidine, putrescine, methylene blue, proline, sugars, glycerol, surfactants, arginine, glycine, guanidine hydrochloride, urea, sodium chloride, potassium chloride, triethylamine, ethanolamine, triethanolamine, ethylenediamine, poloxamer, benzathine penicillin, procaine, lidocaine, bupivacaine, ropivacaine, oxytetracycline, sunitinib, rhizobactin, benzofuran, magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium trisilicate, zinc carbonate, zinc hydroxide, zinc phosphate, aluminum hydroxide, aluminum phosphate, dihydroxyaluminum aminoacetate, calcium phosphate, calcium hydroxide, magnesium aluminum hydroxide, and benzofuran derivatives.

[0043] More specifically, the bioavailability enhancer may be at least one selected from the group consisting of sodium decanoate, disodium hydrogen phosphate, choline, histidine, HEPES, glycerol, surfactants such as Tween 80 and Span 80, arginine, poloxamer, benzathine penicillin and bupivacaine, but is not limited thereto.

[0044] More specifically, the bioavailability enhancer may be at least one selected from the group consisting of sodium decanoate, disodium hydrogen phosphate, poloxamer F127, and benzathine penicillin, but is not limited thereto.

[0045] In a specific embodiment, the content of the bioavailability enhancer may be 2.5 wt % to 250 wt %, 2.5 wt % to 200 wt %, 2.7 wt % to 150 wt % or 2.7 wt % to 100 wt % based on the weight of semaglutide, but is not limited thereto.

[0046] The area under the whole blood concentration-time curve (AUC 总 ), the pharmaceutical composition according to the present disclosure has a blood concentration-time area under the curve (AUC) of semaglutide up to 24 hours after administration of 20% or less, 10% or less, 5% or less, 0.1% to 20%, 1% to 10%, or 1% to 5%. 0-24h ).

[0047] The area under the whole blood concentration-time curve (AUC 总 ), the pharmaceutical composition according to the present disclosure may have a blood concentration-time area under the curve (AUC) of semaglutide from administration to the inter-administration day of 95% or less, 90% or less, 85% or less, 0.1% to 95%, 1% to 95%, 10% to 95%, 1% to 90%, 10 to 90%, 20% to 95%, 30% to 95%, 40% to 95%, 50% to 95%, 60% to 95%, 0% to 95%, 80% to 95%, or 70% to 85% of the blood concentration-time area under the curve (AUC) of semaglutide from administration to the inter-administration day 0-QXM ).

[0048] Furthermore, the pharmaceutical composition according to the present disclosure may be a pharmaceutical composition containing sustained-release microspheres, wherein the content of semaglutide is 12 wt % or more as semaglutide based on the total weight of the microspheres, and the initial release of the drug is less than 10% within 24 hours.

[0049] The biodegradable polymer contained in the semaglutide sustained-release microspheres contained in the pharmaceutical composition according to the present disclosure is at least one selected from the group consisting of polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA) which is a copolymer of lactide and glycolide, polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide- Polymers in the group consisting of poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), poly(vinyl alcohol), and poly(vinyl alcohol).

[0050] In one specific aspect, the pharmaceutical composition according to the present disclosure may include microspheres containing two or more types of the above-mentioned biodegradable polymers. In another specific embodiment, the pharmaceutical composition according to the present disclosure may contain two or more types of microspheres, each microsphere containing two or more types of polymers selected from the above-mentioned biodegradable polymers.

[0051] Two or more polymers selected from the group consisting of polylactide-co-glycolide and polylactide polymers have an intrinsic viscosity of 0.16 dL / g to 1.7 dL / g, 0.2 dL / g to 1.3 dL / g, or 0.24 dL / g to 1.2 dL / g, but are not limited thereto.

[0052] The intrinsic viscosity of polylactide-co-glycolide or polylactide used in the present disclosure refers to the intrinsic viscosity measured at a concentration of 0.1% (w / v) in chloroform using an Ubbelohde viscometer at 25°C. If the intrinsic viscosity of polylactide-co-glycolide or polylactide is less than 0.16dL / g, the molecular weight of the polymer is insufficient, and it is difficult to show the sustained-release effect of semaglutide or its pharmaceutically acceptable salt. If the intrinsic viscosity exceeds 1.7dL / g, it may show an effect of excessively delaying the release of semaglutide or its pharmaceutically acceptable salt. In addition, when microspheres are prepared using a polymer with a high intrinsic viscosity, there is a problem in that, due to the high viscosity of the polymer, an excessive amount of preparation solvent must be used, and it is difficult to prepare reproducible microspheres. Examples of commercially available polymers having the above properties include RG502H, RG503H, RG504H, RG502, RG503, RG504, RG653H, RG752H, RG752S, RG755S, RG750S, RG757S, RG858S, R202H, R203H, R205H, R202S, R203S, R205S, R206S and R207S ( series, purchased from Evonik), and PDL 02A, PDL 02, PDL 04, PDL 05, PDLG 7502A, PDLG 7502, PDLG 7507, PDLG5002A, PDLG 5002, PDLG 5004A and PDLG 5004 (purchased from Corbion Purac), etc.

[0053] The content of the biodegradable polymer in the sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the present disclosure may be 60 wt % to 95 wt %, 65 wt % to 93 wt % or 70 wt % to 90 wt % based on the total weight of the microspheres, but is not limited thereto.

[0054] The content of semaglutide or its pharmaceutically acceptable salt in the sustained-release microspheres containing semaglutide or its pharmaceutically acceptable salt and a bioavailability enhancer disclosed herein is preferably 8 wt% or more, 12 wt% or more, 14 wt% or more, 37 wt% or less, 35 wt% or less, or 33 wt% or less based on the total weight of the microspheres (in terms of semaglutide). If the content of semaglutide or its pharmaceutically acceptable salt in the microspheres is less than 8 wt% (in terms of semaglutide), the amount of polymer used is too high relative to the drug, which may reduce the bioavailability of semaglutide or its pharmaceutically acceptable salt. If the content is too high, there is a problem that the initial release of semaglutide or its pharmaceutically acceptable salt may increase, which is not preferred.

[0055] The microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the present disclosure preferably have a uniform particle distribution curve with 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, The term "average particle size" as used herein refers to the median diameter of the particle size corresponding to 50% of the volume % in the particle size distribution curve, and is represented by D50 or D(v,0.5).

[0056] If the average particle size of the microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer is less than 5 μm, semaglutide or a pharmaceutically acceptable salt thereof is released too quickly from the microspheres, which is not preferred. If the average particle size is greater than 100 μm, the injection needle becomes too thick when administered in the human body, which causes pain during injection, or the drug leaks at the injection site after injection, which is not preferred.

[0057] Preferably, the microparticles containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer disclosed herein have a uniform particle size distribution. Compared with microspheres with uneven particle size distribution, microparticles containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer with uniform particle size distribution can be administered with more precise amount and smaller deviation during injection. Preferably, the span value of the microspheres containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer disclosed herein is 1.5 or less. More preferably, the span value is 1.2 or less. More specifically, the span value can 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 span value used herein is an index representing the uniformity of the particle size of the microspheres, and refers to a value determined by the formula span value = (Dv0.9-Dv0.1) / Dv0.5. Here, Dv0.1 represents a particle size corresponding to 10% of the volume % in the particle size distribution curve of the microspheres, Dv0.5 represents a particle size corresponding to 50% of the volume % in the particle size distribution curve of the microspheres, and Dv0.9 represents a particle size corresponding to 90% of the volume % in the particle size distribution curve of the microspheres.

[0058] Since the sustained-release microspheres containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer disclosed in the present invention are administered by injection (e.g., subcutaneous injection) and can be particularly self-administered, it is preferred that semaglutide is released over a relatively long period of time. Preferably, the sustained-release microspheres in the pharmaceutical composition according to the present invention can release semaglutide or its pharmaceutically acceptable salt for 1 month or longer, 2 months or longer, 3 months or longer, 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, but not limited thereto. In addition, the sustained-release microspheres containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer disclosed in the present invention are not particularly limited to the release mode, but when administered in vivo, it is preferred that less than 10%, less than 15% or less than 20% of semaglutide or its pharmaceutically acceptable salt is released within 24 hours.

[0059] In addition, in the pharmaceutical composition of the present disclosure, the total amount of the sustained-release microsphere preparation containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer can be 20 mg to 1,000 mg, 20 mg to 800 mg, 20 mg to 600 mg, 20 mg to 400 mg, 20 mg to 200 mg, 20 mg to 100 mg, 30 mg to 1,000 mg, 30 mg to 800 mg, 30 mg to 600 mg, 30 mg to 4 00mg, 30mg to 200mg, 30mg to 100mg, 40mg to 1,000mg, 40mg to 800mg, 40mg to 600mg, 40mg to 400mg, 40mg to 200mg, 40mg to 100mg, 50mg to 1,000mg, 50mg to 800mg, 50mg to 600mg, 50mg to 400mg, 50mg to 200mg or 50mg to 100mg. Since the sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer in the composition are included within the above range, it has the following advantages: the composition according to the present disclosure not only reduces the inflammatory response at the application site as much as possible, but can also be administered by the patient himself.

[0060] The microspheres containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer contained in the composition of the present disclosure may further include a release control agent. The material used as the release control agent may include, for example, at least one selected from the group consisting of butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, nonanoic acid, capric acid, undecylenic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, 19-decanoic acid, behe acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, naphthalene dicarboxylic acid, naphthalenesulfonic acid and pamoic acid, but is not limited thereto. Preferably, the release control agent may be hydroxynaphthoic acid, naphthalene dicarboxylic acid or pamoic acid, but is not limited thereto.

[0061] According to the pharmaceutical composition of the present disclosure comprising microspheres containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer, various forms of preparations can be formulated, for example, it can be in the form of known parenteral preparations. Therefore, according to the pharmaceutical composition of the present disclosure, in addition to semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer, thickeners, stabilizers, isotonic agents, initial release inhibitors, surfactants, excipients and / or carriers can also be included. Isotonic agents that can be used can be water-soluble excipients or sugars, such as mannitol, sucrose, sorbitol, trehalose, lactose, sodium chloride, etc., and examples of thickeners include sodium carboxymethyl cellulose (carmellose sodium), sodium carboxymethyl cellulose (carboxymethyl cellulose sodium) and povidone. In addition, disodium hydrogen phosphate, anhydrous citric acid, sodium hydroxide, sodium chloride, etc. can be used as buffers.

[0062] The pharmaceutical composition containing sustained-release microspheres according to the present disclosure may contain less than a certain amount of an initial release inhibitor in the microspheres.

[0063] In the present disclosure, the initial release inhibitor is a substance contained in the continuous phase during the preparation of microspheres in order to inhibit the rapid release of the active ingredient, and is characterized in that it is present in less than a certain amount in the microspheres of the present disclosure.

[0064] In a specific embodiment, the content of the initial release inhibitor may be 5 ppm to 2000 ppm, preferably 10 ppm to 1500 ppm, more preferably 20 ppm to 1,000 ppm, most preferably 20 ppm to 500 ppm based on the total weight of the microspheres.

[0065] In a specific embodiment, the initial release inhibitor is a material that can maintain the pH of the continuous phase at 7 or higher when dissolved in the continuous phase, more specifically, 7 or higher, 7.2 or higher, 7.4 or higher, or 8.0 or higher, 8.5 or higher, 9.0 or higher, 7.0 to 9.0, or 7.0 to 8.5, and any material can be used as long as it can form a divalent or higher valent anion in the continuous phase. The initial release inhibitor can be at least one substance selected from alkali metals, alkaline earth metals or ammonium phosphate salts, phosphide salts, carbonates, chromates, dichromates, oxides, oxalates, silicates, sulfates, sulfide salts, sulfites, tartrates, tetraborates, thiosulfates, arsenates, arsenites, citrates, ferrocyanide salts and nitride salts, but is not limited thereto. In a specific embodiment, the initial release inhibitor can be at least one substance selected from disodium hydrogen phosphate, dipotassium hydrogen phosphate and diammonium hydrogen phosphate, but is not limited thereto.

[0066] The pharmaceutical composition according to the present disclosure can be administered in a therapeutically effective amount of semaglutide, for example, in an amount effective for treating diabetes (particularly type 2 diabetes), β-cell function protection, hypertension, hyperlipidemia, obesity, non-alcoholic fatty hepatitis or neurodegenerative diseases (such as Alzheimer's disease and Parkinson's disease). The therapeutically effective amount of semaglutide can be assessed by a doctor. The pharmaceutical composition containing semaglutide disclosed in the present disclosure can be administered once a month to once a quarter. In some embodiments, the monthly dose of the composition according to the present disclosure can be 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 6mg, 4mg to 100mg, 4mg to 80mg, 4mg to 60mg, 4mg to 30mg, 4mg to 20mg, 8mg to 100mg, 8mg to 80mg, 8mg to 60mg, 8mg to 30mg, 10mg to 100mg, 10mg to 80mg, 10mg to 60mg, 10mg to 30mg, 20mg to 100mg, 20mg to 80mg, 20mg to 60mg or 20mg to 30mg. The pharmaceutical composition comprising microspheres according to the present disclosure can be administered parenterally, for example by subcutaneous injection, and the microspheres contain semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the present disclosure. The pharmaceutical composition according to the present disclosure can be composed of a drug portion containing microspheres and a solvent portion for suspending the microspheres, and the microspheres contain semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer. In addition, the pharmaceutical composition may be in the form of a dual-chamber syringe containing a drug portion in one chamber and a solvent portion in another chamber, or in the form of a pre-filled syringe in which the drug portion is suspended in the solvent portion. When the drug portion is configured as a pre-filled syringe in which the drug portion is suspended in the solvent portion, the solvent portion used may be an injectable oil containing a medium-chain oil, mineral oil, or the like.

[0067] In a specific embodiment, the sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer contained in the pharmaceutical composition according to the present disclosure have a high drug content compared to the content of the microspheres, inhibit the initial excessive release of the drug that can cause fatal side effects, and have high bioavailability, showing sufficient efficacy as a GLP-1 agonist within the required time, and can therefore be used to prevent or treat diabetes (especially type 2 diabetes), protection of β-cell function, hypertension, hyperlipidemia, obesity, non-alcoholic fatty hepatitis or neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease.

[0068] In another aspect of the present invention, a method for preparing sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer is provided.

[0069] In another aspect of the present invention, a method for producing sustained-release microspheres having significantly suppressed initial release and containing a high content of semaglutide or a pharmaceutically acceptable salt thereof is provided.

[0070] The preparation method of the sustained-release microsphere injection containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer disclosed in the present invention will be described in detail below.

[0071] The sustained-release microsphere injection formulation containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the present disclosure can be prepared using, for example, a “solvent extraction and evaporation method”, but is not limited thereto.

[0072] In one aspect of the specific preparation method disclosed herein, a method for preparing sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer is provided, comprising the following steps:

[0073] (a) a step of dissolving semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and one or more biodegradable polymers in one or more organic solvents to prepare a solution (dispersed phase) containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer;

[0074] (b) a step of adding the solution containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer prepared in step (a) to an aqueous solution phase (continuous phase) containing a surfactant to prepare an emulsion;

[0075] (c) extracting an organic solvent from the dispersed phase in the emulsion state prepared in step (b) into a continuous phase, and evaporating the organic solvent to form microspheres; and

[0076] (d) recovering microspheres from the continuous phase of step (c) to prepare microspheres containing semaglutide or a pharmaceutically acceptable salt thereof with enhanced bioavailability and a bioavailability enhancer.

[0077] In another aspect, the bioavailability enhancer in step (a) is at least one selected from the group consisting of: poloxamer, benzathine penicillin, procaine, lidocaine, bupivacaine, ropivacaine, oxytetracycline, sunitinib, rhizobactin and benzofuran.

[0078] In one aspect of the present disclosure, there is provided a method for preparing sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer, comprising the following steps:

[0079] (a') dissolving semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer in an aqueous solution to prepare a primary aqueous solution phase, dissolving one or more biodegradable polymers in an organic solvent to prepare an oil phase, and then mixing the primary aqueous solution and the oil phase to prepare a W / O emulsion (primary emulsion) containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer;

[0080] (b') adding the primary emulsion containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer prepared in step (a') to an aqueous solution phase (continuous phase) containing a surfactant to prepare a W / O / W emulsion (secondary emulsion);

[0081] (c') extracting an organic solvent from the oil phase in the secondary emulsion state prepared in step (b') into a continuous phase, and evaporating the organic solvent to form microspheres; and

[0082] (d') recovering the microspheres from the continuous phase of step (c') to prepare microspheres containing semaglutide or a pharmaceutically acceptable salt thereof with enhanced bioavailability and a bioavailability enhancer.

[0083] In another aspect, the bioavailability enhancer in step (a') is at least one selected from the group consisting of sodium caprate, disodium hydrogen phosphate, choline, meglumine, basic aluminum carbonate, sodium dihydroxyaluminum carbonate, ammonium phosphate, histidine, HEPES, HEPPS, spermine, spermidine, putrescine, methylene blue, proline, sugars, glycerol, surfactants, arginine, glycine, guanidine hydrochloride, urea, sodium chloride, potassium chloride, triethyleneamine, ethanolamine, triethanolamine and ethylenediamine.

[0084] In another aspect of the present disclosure, there is provided a method for preparing sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer, comprising the following steps:

[0085] (a”) dissolving semaglutide or a pharmaceutically acceptable salt thereof and one or more biodegradable polymers in an organic solvent, and suspending a bioavailability enhancer in the solution to prepare a suspension (dispersed phase) containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer;

[0086] (b") adding the suspension containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer prepared in step (a") to an aqueous solution phase (continuous phase) containing a surfactant to prepare an emulsion;

[0087] (c") extracting an organic solvent from the dispersed phase in the emulsion state prepared in step (b") into a continuous phase, and evaporating the organic solvent to form microspheres; and

[0088] (d") recovering the microspheres from the continuous phase of step (c") to prepare microspheres containing semaglutide or a pharmaceutically acceptable salt thereof with enhanced bioavailability and a bioavailability enhancer.

[0089] On the other hand, the bioavailability enhancer in step (a") is at least one selected from the group consisting of magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium trisilicate, zinc carbonate, zinc hydroxide, zinc phosphate, aluminum hydroxide, aluminum phosphate, dihydroxyaminoacetic acid aluminum, calcium phosphate, calcium hydroxide, magnesium aluminum hydroxide and benzofuran derivatives.

[0090] On the other hand, the pH of the continuous phase used in the above-mentioned preparation method may be 7 or higher.

[0091] In the preparation of sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the present disclosure, in order to suppress the initial excessive release of semaglutide or a pharmaceutically acceptable salt thereof while containing a high content of semaglutide or a pharmaceutically acceptable salt thereof relative to the weight of the microparticles, having high bioavailability, and a long period of time required for release at a constant concentration, for example, 1 month or longer, 3 months or longer, 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, the following biodegradable polymers may be used, but are not limited thereto. The biodegradable polymer that can be used can be at least one polymer selected from the group consisting of polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA) as a copolymer of lactide and glycolide, polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co- -trimethylene carbonate (PLTMC), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoesters, polyanhydrides, polyanhydride-co-imides, polypropylene fumarate, pseudo-polyamino acids, polyalkyl cyanoacrylates, polyphosphazenes, polyphosphates, poly(butylene succinate / lactide) (PBSLA); simple mixtures of two or more thereof; copolymers of the above polymers with polyethylene glycol (PEG); and polymer-sugar conjugates in which the polymer or copolymer is conjugated to a sugar. In a specific embodiment, in the preparation method according to the present disclosure, poly(lactide-co-glycolide) and / or polylactide polymers can be used as biodegradable polymers.

[0092] In the present disclosure, the two or more different types of biodegradable polymers may include two or more polymers having different repeating units constituting the polymers and two or more polymers having different repeating unit molar ratios when containing two or more repeating units.

[0093] As an example, the microspheres may be a mixture of microspheres containing polylactide-co-glycolide and microspheres containing a polylactide polymer, or may be microspheres containing polylactide-co-glycolide and a polylactide polymer.

[0094] In addition, as a specific example, when two types of biodegradable polymers different from each other are used, the content ratio of these biodegradable polymers can be 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, but is not limited thereto.

[0095] In a more specific embodiment, in order to prepare sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer according to the present disclosure, when polylactide-co-glycolide is used as two or more types of biodegradable polymers, at least one biodegradable polymer having a viscosity of 0.16 dL / g to 0.45 dL / g may be included.

[0096] In step (a), (a') or (a") of the above-mentioned specific preparation method, the organic solvent for dissolving the one or more biodegradable polymers is one or more organic solvents. In addition, as the organic solvent, a mixed organic solvent in which two or more types of organic solvents are mixed can also be used. In a specific embodiment, the mixed solvent can be a mixed solvent of an organic solvent miscible with water and an organic solvent immiscible with water. In this case, the organic solvent immiscible with water is preferably present in an amount of at least 50% (v / v) or more, 60% (v / v) or more. % or more, 50 (v / v)% to 99.9 (v / v)%, 50 (v / v)% to 90 (v / v)%, 50 (v / v)% to 80 (v / v)%, 50 (v / v)% to 70 (v / v)%, 60 (v / v)% to 90 (v / v)% or 60 (v / v)% to 80 (v / v)%. By using the water-immiscible property of the organic solvent, the continuous phase containing the surfactant in the following step (b), (b') or (b") can be uniformly mixed with the dispersed phase to form an emulsion. The type of organic solvent in which one or more of these biodegradable polymers are dissolved 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, methanol, ethanol, propanol and benzyl alcohol, more preferably a solvent selected from dichloromethane and ethyl acetate, and one or more organic solvents selected from dimethyl sulfoxide, N-methylpyrrolidone, methanol and acetic acid can be used.

[0097] The method for uniformly mixing the dispersed phase and the continuous phase containing the surfactant in step (b), (b') or (b") is not particularly limited, but can be carried out using a high-speed stirrer, an in-line mixer, a membrane emulsification method, a microfluidic emulsification method, an ultrasonic mixer, a static mixer, etc. When an emulsion is formed using a high-speed stirrer, an in-line mixer, an ultrasonic mixer or a static mixer, it is difficult to obtain a uniform emulsion, and therefore it is preferred to perform an additional sieving step between steps (c) and (d), between steps (c') and (d'), or between steps (c") and (d"), which will be described below.

[0098] The type of surfactant used in step (b), (b') or (b") is not particularly limited, and any surfactant can be used as long as it can help the dispersed phase form a stable droplet dispersion phase within the continuous phase. Polyvinyl alcohol can be used as the surfactant.

[0099] In step (b), (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 containing the surfactant.

[0100] If the surfactant content 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 if the surfactant content exceeds 20 w / v%, fine particles are formed in the continuous phase due to excess surfactant, and it may be difficult to remove the surfactant.

[0101] The continuous phase used in step (b), (b') or (b") may be water, and in order to control the initial release of the microspheres having a high content of drug encapsulated therein, a substance that forms multivalent anions when dissolved in water may be further included as an initial release inhibitor.

[0102] In step (b), (b') or (b"), the type of the initial release inhibitor is not particularly limited and can be used without limitation as long as it is any basic salt that can be dissolved in the continuous phase and maintain the pH at 7.0 or higher. As the initial release inhibitor, at least one substance selected from alkali metal, alkaline earth metal or ammonium phosphate salts, phosphide salts, carbonates, chromates, dichromates, oxides, oxalates, silicates, sulfates, sulfide salts, sulfites, tartrates, tetraborates, thiosulfates, arsenates, arsenites, citrates, ferrocyanide salts and nitride salts can be used.

[0103] In a specific embodiment, the initial release inhibitor may be at least one substance selected from the group consisting of disodium hydrogen phosphate, dipotassium hydrogen phosphate, and diammonium hydrogen phosphate, but is not limited thereto.

[0104] In step (b), (b') or (b"), the content of the initial release inhibitor contained in the continuous phase used may be 0.05 w / v% to 20 w / v%, preferably 0.1 w / v% to 15 w / v%, more preferably 0.2 w / v% to 12.5 w / v%, and most preferably 0.3 w / v% to 10 w / v%, based on the total volume of the continuous phase containing the initial release inhibitor.

[0105] If the content of the initial release inhibitor exceeds 20 w / v%, emulsion droplet breakage may occur during microsphere preparation. On the contrary, if the content is less than 0.05 w / v%, the initial release of microspheres containing a high content of semaglutide may not be sufficiently inhibited.

[0106] When an initial release inhibitor is included in the continuous phase during preparation of microspheres, the initial release from microspheres containing 12 (w / w)% or more semaglutide can be reduced to preferably 10% or less, more preferably 8% or less, most preferably 5% or less.

[0107] The continuous phase used in step (b), (b') or (b") may be water, and in order to adjust the extraction rate of the organic solvent from the dispersed phase in the emulsion state, water containing a portion of one or more selected from the group consisting of methanol, ethanol, propanol and ethyl acetate may be used.

[0108] In addition, the pH of the continuous phase may be 7.0 or higher, 7.2 or higher, 7.4 or higher, 8.0 or higher, 8.5 or higher, 9.0 or higher, 7.0 to 9.0, or 7.0 to 8.5, but is not limited thereto. When the pH of the continuous phase is adjusted within the above range, the bioavailability of the microspheres containing semaglutide or a pharmaceutically acceptable salt thereof can be further increased.

[0109] In step (c), (c') or (c"), when the emulsion containing a dispersed phase in the form of droplets and a continuous phase containing a surfactant is maintained or stirred at a temperature below the boiling point of the organic solvent for a certain period of time, for example, 2 hours to 48 hours, the organic solvent can be extracted from the semaglutide or its pharmaceutically acceptable salt-bioavailability enhancer-polymer solution in the form of droplets as the dispersed phase into the continuous phase. Part of the organic solvent extracted into the continuous phase can evaporate from the surface of the continuous phase. Since the organic solvent is extracted and evaporated from the semaglutide or its pharmaceutically acceptable salt-bioavailability enhancer-polymer solution in the form of droplets, the dispersed phase in the form of droplets can be solidified to form microspheres.

[0110] In step (c), (c') or (c"), in order to further effectively remove the organic solvent, the temperature of the continuous phase may be heated for a certain time. The heating temperature is not limited and may be appropriately adjusted by a person skilled in the art according to the organic solvent used. For example, when dichloromethane is used as the organic solvent, heat may be applied to maintain the temperature at 30°C or higher, 40°C or higher, 45°C or higher, 30°C to 50°C, 40°C to 50°C, or 45°C.

[0111] In step (d), (d') or (d"), the method of recovering the microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer can be carried out using several known techniques, and for example, methods such as filtration or centrifugation can be used.

[0112] Between steps (c) and (d), between steps (c') and (d'), or between steps (c") and (d"), residual surfactant may be removed by filtering and washing, and then filtering may be performed again to recover the microspheres.

[0113] The washing step for removing residual surfactant may generally be performed using water, and the washing step may be repeated several times.

[0114] In addition, as described above, uniform microspheres can be obtained by additionally using a sieving step between steps (c) and (d) and between steps (c') and (d') or between steps (c") and (d"). The sieving step can be performed using known techniques, and sieving membranes with different sizes can be used to filter microspheres of small and large particles to obtain microspheres of uniform size.

[0115] In the preparation method of the present disclosure, after step (d), step (d') or step (d"), or after filtering and washing steps, the obtained microspheres are dried using a conventional drying method to finally obtain dried microspheres.

[0116] For all substances not separately defined other than those described above, including semaglutide, bioavailability enhancers, biodegradable polymers, and contents thereof, the substances defined in the pharmaceutical composition may be used as they are.

[0117] According to the preparation method disclosed in the present invention, a sustained-release microsphere injection preparation containing semaglutide or a pharmaceutically acceptable salt thereof with high bioavailability can be prepared, wherein the drug is maintained at an effective concentration for a desired period of time without rapid short-term release. In addition, a sustained-release microsphere injection preparation containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer in the form of uniform particles with good applicability can be prepared.

[0118] Example

[0119] The present disclosure will be described in more detail below through preparation examples. However, these preparation examples are only for illustrative purposes, and the scope of the present disclosure is not limited to the preparation examples.

[0120] Preparation Example

[0121] Preparation of biodegradable polymer microspheres containing semaglutide or its pharmaceutically acceptable salt and bioavailability enhancer

[0122] Preparation Example 1 (w / o / w method): containing semaglutide and a bioavailability enhancer dissolved in an internal aqueous phase Preparation of biodegradable polymer microspheres

[0123] 0.15g of semaglutide sodium salt as a drug (manufacturer: Chengdu Kaijie Biopharmaceutical Co., Ltd., China) and 0.33g of poloxamer F127 as a bioavailability enhancer were dissolved in 1.66g of tertiary distilled water to prepare the primary aqueous phase. RG653H and RG753H (manufacturer: Evonik, Germany) as biodegradable polymers were mixed into 1.52g in a weight ratio of 1:1 and dissolved in 20.8g of dichloromethane (manufacturer: JTBaker, USA) to prepare the organic phase. The aqueous phase was dispersed in the organic phase using a homogenizer to prepare a primary emulsion (dispersed phase). As the continuous phase, 2,000mL of 0.1% polyvinyl alcohol aqueous solution (viscosity: 4.8mPa·s to 5.8mPa·s) was used. The continuous phase was connected to an emulsifying device equipped with a porous membrane with a diameter of 40μm, and the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion having dispersed biodegradable polymer droplets containing semaglutide was prepared, and the suspension was placed in a preparation container and stirred at a speed of 200 rpm.

[0124] The temperature of the preparation container was maintained at 25° C., and when the injection of the dispersed phase was completed, the organic solvent was removed while the suspension temperature was maintained at 40° C. for 3 hours. Subsequently, after the temperature was cooled to 25° C., the microspheres were filtered, treated with tertiary distilled water to remove residual polyvinyl alcohol, and freeze-dried.

[0125] Preparation Example 2 (w / o / w method): containing semaglutide and a bioavailability enhancer dissolved in an internal aqueous phase Preparation of biodegradable polymer microspheres containing an initial release inhibitor in a continuous phase

[0126] 0.2g of semaglutide sodium salt as a drug and 0.02g of disodium hydrogen phosphate as a bioavailability enhancer were dissolved in 1.0g of tertiary distilled water to prepare a primary aqueous phase. By dissolving 0.8g of PDL 04A (manufacturer: Corbion, the Netherlands) as a biodegradable polymer in 6.65g of dichloromethane. The aqueous phase was dispersed in the organic phase using a homogenizer to prepare a primary emulsion (dispersed phase). As a continuous phase, 2,000mL of 0.1% polyvinyl alcohol aqueous solution containing 2% (w / w) disodium hydrogen phosphate as an initial release inhibitor was used. The continuous phase was connected to an emulsifying device equipped with a porous membrane with a diameter of 40μm, and the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion with dispersed biodegradable polymer droplets containing semaglutide was prepared, and the suspension was placed in a preparation container and stirred at a speed of 200rpm.

[0127] The temperature of the preparation vessel was maintained at 25° C., and when the injection of the dispersed phase was completed, the organic solvent was removed while the suspension temperature was maintained at 40° C. for 3 hours. Subsequently, the temperature was cooled to 25° C., the microspheres were filtered, treated with tertiary distilled water to remove residual polyvinyl alcohol, and freeze-dried.

[0128] Preparation Example 3 (w / o / w method): containing semaglutide dissolved in the internal aqueous phase, a release control agent and a biopharmaceutical Preparation of biodegradable polymer microspheres containing an availability enhancer and an initial release inhibitor in a continuous phase

[0129] 0.2 g of semaglutide free base as a drug (manufacturer: Chengdu Kaijie Biopharmaceutical Co., Ltd., China) and 0.02 g each of sodium decanoate and disodium hydrogen phosphate as release control agents and bioavailability enhancers were dissolved in 1.8 g of tertiary distilled water to prepare a primary aqueous phase. The organic phase was prepared by dissolving 0.76 g of PDLG 7504A (manufacturer: Corbion, the Netherlands) as a biodegradable polymer in 14.4 g of dichloromethane. The aqueous phase was dispersed in the organic phase using a homogenizer to prepare a primary emulsion (dispersed phase). As a continuous phase, 2,000 mL of 0.1% polyvinyl alcohol aqueous solution containing 2% (w / w) disodium hydrogen phosphate as an initial release inhibitor was used. The continuous phase was connected to an emulsifying device equipped with a porous membrane with a diameter of 40 μm, and the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion having dispersed biodegradable polymer droplets containing semaglutide was prepared, and the suspension was placed in a preparation container and stirred at a speed of 200 rpm.

[0130] The temperature of the preparation vessel was maintained at 25° C., and when the injection of the dispersed phase was completed, the organic solvent was removed while the suspension temperature was maintained at 40° C. for 3 hours. Subsequently, the temperature was cooled to 25° C., the microspheres were filtered, treated with tertiary distilled water to remove residual polyvinyl alcohol, and freeze-dried.

[0131] Preparation Example 4 (o / w method): containing semaglutide and a bioavailability enhancer dissolved in an organic phase and Preparation of biodegradable polymer microspheres containing an initial release inhibitor in a continuous phase

[0132] By mixing 0.79 g of PDLG 7504A as a biodegradable polymer, 0.2 g of semaglutide free base as a drug, and 0.006 g of benzathine penicillin as a bioavailability enhancer, and using 19.9 g of dichloromethane, glacial acetic acid (manufacturer: Daejeong, South Korea) and methanol (manufacturer: Daejeong, South Korea) (weight ratio of about 16:4:1) The mixture is stirred thoroughly for 30 minutes or more. As for the continuous phase, 2,000 mL of 0.1% polyvinyl alcohol aqueous solution containing 2% (w / w) disodium hydrogen phosphate as an initial release inhibitor is used. The continuous phase is connected to an emulsifying device equipped with a porous membrane with a diameter of 40 μm, and the prepared dispersed phase is injected into the porous membrane together with the continuous phase. An emulsion with dispersed biodegradable polymer droplets containing semaglutide is prepared, and the suspension is placed in a preparation container and stirred at a speed of 200 rpm.

[0133] The temperature of the preparation vessel was maintained at 25° C., and when the injection of the dispersed phase was completed, the organic solvent was removed while the suspension temperature was maintained at 40° C. for 3 hours. Subsequently, the temperature was cooled to 25° C., the microspheres were filtered, treated with tertiary distilled water to remove residual polyvinyl alcohol, and freeze-dried.

[0134] Preparation Examples 5 to 32: Biodegradable polymers containing semaglutide or a pharmaceutically acceptable salt thereof Preparation of bioparticle microspheres

[0135] 0.125 to 0.450 g of semaglutide (Manufacturer: Chengdu Kaijie Biopharmaceutical Co., Ltd., China) as the drug and 0 g to 1.2 g of the bioavailability enhancer (see Table 1) were used, and finally, at least one of R203H, RG203H, RG503H, RG653H, RG753H, PDL04A and PDLG7504A ranging from 0.75 g to 2.55 g of the biodegradable polymer was weighed so that the batch size was 1.0 g to 3.0 g (in Preparation Examples 1 to 32, the batch size of Preparation Example 16, Preparation Example 19 and Preparation Example 30 was 3.0 g, the batch size of Preparation Example 1, Preparation Example 13, Preparation Example 14, Preparation Example 15, Preparation Example 29 and Preparation Example 30 was 2.0 g, and the batch size of the remaining part was 1.0 g). Dichloromethane was used as a solvent to prepare a dispersed phase, and at least one of glacial acetic acid and methanol was used as a co-solvent to uniformly dissolve Preparation Examples 4 to 29 and Preparation Examples 31 and 32. The aqueous phase was dispersed in the organic phase using a 40 μm porous emulsion membrane to prepare a primary emulsion. In Preparation Examples 2 to 32, 2,000 ml of an aqueous solution containing 0.1% polyvinyl alcohol (viscosity: 4.8 mPa.s to 5.8 mPa.s) and 2.0% Na2HPO4 (disodium hydrogen phosphate) were used as a continuous phase, and in Preparation Example 1, 2,000 ml of an aqueous solution containing polyvinyl alcohol (viscosity: 4.8 mPa.s to 5.8 mPa.s) and 1.0% NaCl were used. The continuous phase was connected to an emulsifying device equipped with a porous membrane having a diameter of 40 μm, and the prepared dispersed phase was injected into the porous membrane together with the continuous phase. An emulsion having dispersed biodegradable polymer droplets containing semaglutide was prepared, and the suspension was placed in a preparation container and stirred at a speed of 200 to 300 rpm.

[0136] The temperature of the preparation vessel was maintained at 25° C., and when the injection of the dispersed phase was completed, the organic solvent was removed while the suspension temperature was maintained at 40° C. for 3 hours. Subsequently, the temperature was cooled to 25° C., the microspheres were filtered, treated with tertiary distilled water to remove residual polyvinyl alcohol, and freeze-dried.

[0137] Preparation Examples 5 and 10 did not contain an initial release inhibitor in the continuous phase.

[0138] [Table 1]

[0139]

[0140]

[0141]

[0142]

[0143] a) semaglutide sodium salt, b) semaglutide free base

[0144] c) O / W method, d) W / O / W method

[0145] Preparation Examples 1 to 4 show the preparation of microspheres according to different bioavailability enhancers by using poloxamer F127, disodium hydrogen phosphate, sodium decanoate and disodium hydrogen phosphate, and benzathine penicillin as bioavailability enhancers, respectively. Preparation Examples 4 to 9 show the preparation of microspheres according to the content of benzathine penicillin, Preparation Examples 11 to 15 show the preparation of microspheres according to the content of semaglutide, and Preparation Examples 16 to 29 show the preparation of microspheres according to polymers.

[0146] Preparation Examples 33 to 37 (O / W method): A pharmaceutically acceptable salt containing dexamethasone acetate or a pharmaceutically acceptable salt thereof Preparation of biodegradable polymer microspheres

[0147] For the preparation of dexamethasone microspheres, the dispersed phase includes at least one of Purasorb PDLG7502A (intrinsic viscosity 0.16 dl / g to 0.24 dl / g; manufacturer: Purac, the Netherlands), RG 752H (intrinsic viscosity 0.14 dl / g to 0.22 dl / g; manufacturer: Evonik, Germany) and RG 753H (intrinsic viscosity 0.32 dl / g to 0.44 dl / g; manufacturer: Evonik, Germany) as biocompatible polymers, PDL04A, PDL02, PDLG7504A (manufacturer: Corbion, the Netherlands), 0.200 g to 0.400 g of dexamethasone acetate (manufacturer: Pfizer, the United States) is mixed with dichloromethane (manufacturer: JT Baker, the United States), dimethyl sulfoxide (manufacturer: JT Baker, the United States) and benzyl alcohol (manufacturer: Junsei, Japan) to prepare microspheres. In Preparation Examples 33 to 37, 2,000 ml of an aqueous solution containing 0.5% polyvinyl alcohol (viscosity: 4.8 mPa.s to 5.8 mPa.s) and 2.5% NaCl was used as a continuous phase.

[0148] [Table 2]

[0149]

[0150] Preparation Examples 33 to 37 show the preparation of microspheres according to the type of polymer and the amount of dexamethasone acetate used.

[0151] Preparation Example 38 to Preparation Example 51

[0152] The co-administration formulations of semaglutide microspheres and dexamethasone acetate microspheres are shown in Preparation Examples 38 to 51 (see Table 3).

[0153] [Table 3]

[0154]

[0155]

[0156] Experimental Example 1: Measurement of the amount of semaglutide encapsulated in microspheres

[0157] In order to measure the encapsulation amount of semaglutide in the microspheres prepared in Preparation Examples 1 to 32 and the encapsulation amount of dexamethasone in the microspheres prepared in Preparation Examples 33 to 37, 10 mg of microspheres were completely dissolved in DMSO and then diluted with the mobile phase. 20 μL of the diluted solution was injected into the HPLC and measured at a detection wavelength of 214 nm. The chromatographic column used in this experiment was ZORBAX 300SB-C18, 5 μm, 4.6x150 mm, and the mobile phase was a mixture of 45% 0.1% TFA acetonitrile and 55% 0.1% TFA aqueous solution, using an isocratic mode. The encapsulation amount of the measured drug is shown in Table 4 below.

[0158] [Table 4]

[0159]

[0160]

[0161]

[0162] Experimental Example 2: In vivo pharmacokinetic study using rats

[0163] Experimental Example 2-1: Changes in blood concentration of semaglutide according to bioavailability enhancer and release controller

[0164] In order to evaluate the drug release profile of the semaglutide sustained-release microspheres according to the present disclosure in vivo, the blood concentration of semaglutide was measured after administration to rats.

[0165] The amount of semaglutide was 1.2 mg (4.0 mg / kg) or 3.6 mg (12.0 mg / kg), dispersed in 0.5 mL of suspension, and then injected into SD rats (Sprague-Dawley rats, 300 g) to measure microspheres. 0.5 mL of blood was collected at a pre-planned time, and the blood concentration of semaglutide was determined using HPLC. The measured AUC and C max This is shown in Table 5 below.

[0166] [Table 5]

[0167]

[0168] As can be seen from Table 5, the microspheres containing semaglutide prepared in Preparation Examples 1 to 4 of the present disclosure showed high AUC, and the cumulative AUC ratio from the administration time to 24 hours after administration was less than 10%, which may represent the initial release curve upon administration.

[0169] Experimental Example 2-2. Improvement of semaglutide bioavailability according to benzathine penicillin content

[0170] In order to evaluate the drug release curve of the semaglutide sustained-release microspheres according to the present disclosure in vivo, the bioavailability (%) after administration to rats was measured. Specifically, the content of benzathine penicillin relative to the total weight of the batch was set to 0.6 (w / w)%, 1.2 (w / w)%, 2.3 (w / w)% and 5 (w / w)%. For the semaglutide sustained-release microspheres with added benzathine penicillin and the semaglutide sustained-release microspheres without added benzathine penicillin, 1.2 mg (4.0 mg / kg) or 3.6 mg (12.0 mg / kg) of semaglutide contained in the sustained-release microspheres was dispersed in 0.5 mL of suspension and then injected into SD rats (Sprague-Dawley rats, 300 g). Afterwards, in order to compare the improvement of bioavailability in rats, pharmacokinetic parameters were analyzed. The results confirmed that Preparation Example 4, Preparation Example 7, Preparation Example 8 and Preparation Example 9 showed an AUC increase of about two times or more compared with Preparation Example 6. This confirms that, overall, the AUCs were similar and the bioavailability was 40% or higher.

[0171] [Table 6]

[0172]

[0173] Experimental Example 2-3. Improvement of AUC according to semaglutide content

[0174] In order to evaluate the in vivo drug release profile of the semaglutide sustained-release microspheres according to the present disclosure, AUC was determined after administration to rats.

[0175] In the experiment, semaglutide sustained-release microspheres containing 0.125g, 0.150g, 0.350g and 0.400g of semaglutide and 1.2g of benzathine penicillin as a bioavailability enhancer, and semaglutide sustained-release microspheres containing 0.400g of semaglutide and no bioavailability enhancer were injected into rats. For the injection of semaglutide microspheres, sustained-release microspheres containing 3.6mg (12.0mg / kg) of semaglutide were dispersed in 0.5mL of suspension and then injected into SD rats (Sprague-Dawley rats, 300g).

[0176] [Table 7]

[0177]

[0178] It was confirmed that AUC increased with increasing drug content. In particular, when the microspheres of Preparation Example 12 were administered to experimental animals, according to Figure 1 The results showing the changes in blood drug concentration and pharmacokinetic parameters showed that the cumulative drug release rate within 28 days after administration was 62.28% of the total drug release. These confirmed that the ratio of drug release during drug administration to total drug release was 80% or less, showing a preferred drug release curve.

[0179] Experimental Example 2-4. Initial release according to whether an initial release inhibitor is used

[0180] Table 8 below shows the initial release according to whether the initial release inhibitor is included in the continuous phase.

[0181] [Table 8]

[0182]

[0183]

[0184] The experimental results confirmed that the initial release of the drug from microspheres containing a high content of semaglutide was significantly reduced due to the presence of an initial release inhibitor in the continuous phase during microsphere preparation.

[0185] Experimental Example 3-1. Comparison of microsphere properties by adding disodium hydrogen phosphate and using similar salts

[0186] This experiment was performed using salts such as sodium hydrogen phosphate as initial release inhibitors to compare their properties during the preparation of microparticles.

[0187] A control formulation (Preparation Example 55) without adding the initial release inhibitor in Table 1 and formulations prepared by adding Na2HPO4, K2HPO4, and NaH2PO4 as initial release inhibitors are shown in Preparation Examples 56 to 58. Table 9 shows the main characteristics and initial release of Preparation Examples 55 to 58.

[0188] [Table 9]

[0189] Preparation Example No. Preparation Example 55 Preparation Example 56 Preparation Example 57 Preparation Example 58 Initial release inhibitor N / A <![CDATA[2%Na2HPO4]]> <![CDATA[2% K2HPO4]]> <![CDATA[2%NaH2PO4]]> Initial release (%) 4.6 0.4 0.9 17.0 pH ≒3 ≒7 ≒7 ≒5

[0190] Referring to Preparation Examples 55 to 58 of Table 1 and Table 9, when Na2HPO4 or K2HPO4 was added as an initial release inhibitor, the initial release was reduced to less than 1% compared to the control formulation. However, when NaH2PO4 was added to the continuous phase as an initial release inhibitor, the initial release was 17%, which seemed to be much higher than the initial release of the control formulation. These confirm that even salts similar to disodium hydrogen phosphate do not show the effect of inhibiting the initial release. Preparation Example 55 has about pH 3, Preparation Example 56 and Preparation Example 57 have about pH 7, and Preparation Example 58 has about pH 5. Although no linear data is shown, the reason for the high initial release in Preparation Example 55 and Preparation Example 58 may be considered to be the pH factor. SEM observation of cross-sectional images revealed that a large number of internal pores were distributed in the pH acidic preparations, while the internal pores were reduced in the pH neutral or alkaline preparations. This result is believed to explain the reason for the inhibition of initial release.

[0191] Experimental Example 3-2. Comparison of microsphere characteristics according to the type of initial release inhibitor added

[0192] This experiment was conducted to compare the characteristics due to the type of initial release inhibitor when microspheres were prepared using Na2CO3, NaHCO3, (NH4)2SO4, and (NH4)2HPO4 as the initial release inhibitor.

[0193] The control formulation without adding the initial release inhibitor is shown in Table 1 (Preparation Example 66), and the formulations prepared by adding Na2CO3, NaHCO3, (NH4)2SO4 and (NH4)2HPO4 as the initial release agents are shown in Preparation Examples 68 to 71. The following Table 10 shows the main characteristics and initial release of Preparation Example 66 and Preparation Examples 68 to 71.

[0194] [Table 10]

[0195]

[0196] With reference to Preparation Examples 66, 68 to 71 of Tables 1 and 10, the initial release performance in Preparation Examples 66 and 70 seems to be slightly higher. Preparation Examples 66 and 70 with high initial release show acidic pH values ​​of about 3 and about 5, respectively, and Preparation Examples 68, 69 and 71 with neutral and alkaline pH values ​​show an initial release of less than 3%. Based on these results, when the continuous phase exhibits an acidic pH value by adding an initial release inhibitor to the continuous phase, the initial release of semaglutide seems to be significantly higher. However, when an initial release inhibitor that causes an alkaline pH value to be expressed at pH 7 or higher (neutral or alkaline) is used, the initial release of semaglutide is significantly inhibited. Here, by considering the initial release of Preparation Examples 68, 69 and 71, the initial release is more inhibited in alkaline than in neutral, but the inhibition of the initial release does not increase linearly with the degree of alkalinity.

[0197] Experimental Example 3-3. Comparison of microsphere properties by pH adjustment after adding disodium hydrogen phosphate

[0198] When the pH was intentionally adjusted after adding 2 (w / v)% disodium hydrogen phosphate as an initial release inhibitor, the test results of the initial release change confirmed that the initial release inhibitory effect on the microspheres was maintained even if the pH was changed after the initial release inhibitor was added. Specifically, a control formulation (Preparation Example 66) without adding the initial release inhibitor in Table 1 and a formulation in which 2 (w / v)% disodium hydrogen phosphate was added as an initial release inhibitor but the pH was not intentionally adjusted are shown in Preparation Example 63, a formulation in which HCl was used to adjust the pH to pH 2 (acidic) is shown in Preparation Example 64, and a formulation in which NaOH was used to adjust the pH to pH 12 is shown in Preparation Example 65.

[0199] Table 11 below shows the main characteristics and initial release of Preparation Examples 63 to 66.

[0200] [Table 11]

[0201]

[0202] As shown in Table 11, when microspheres were prepared using a continuous phase containing 2 (w / v)% Na2HPO4 as an initial release inhibitor, the effect of inhibiting the initial release of semaglutide was maintained even when the pH of the continuous phase was subsequently intentionally adjusted to acidic.

[0203] Experimental Example 4. Comparison of microsphere properties according to different dosages of disodium hydrogen phosphate

[0204] This experiment was conducted to compare the microsphere characteristics according to the added amount of disodium hydrogen phosphate as an initial release inhibitor.

[0205] The control formulation (Preparation Example 55) without the addition of Na2HPO4 in Table 1 and the formulations containing 0.5 (w / v)%, 1 (w / v)%, 2 (w / v)%, 3 (w / v)%, and 4 (w / v)% of Na2HPO4 are shown in Preparation Example 59, Preparation Example 60, Preparation Example 56, Preparation Example 61, and Preparation Example 62, respectively. The following Table 12 shows the main characteristics and initial release of Preparation Example 55, Preparation Example 56, and Preparation Examples 59 to Preparation Examples 62.

[0206] [Table 12]

[0207]

[0208] With reference to Preparation Examples 55, 56, and 59 to 62 in Tables 1 and 12, the control formulation had an initial release of 4.6%, while the initial release of the formulation containing 0.5 (w / v)% Na2HPO4 was reduced to 0.64%. When the Na2HPO4 concentration was 1 (w / v)% or 2 (w / v)%, it was confirmed that the initial release rate was reduced to 0.56% and 0.4%, respectively. However, when 3 (w / v)% or 4 (w / v)% Na2HPO4 was added, respectively, the initial release tended to increase to 0.77% and 0.86% compared to when 2% Na2HPO4 was added. When 4 (w / v)% or higher Na2HPO4 was used, a large amount of polymer aggregation occurred during the preparation of microspheres. It was shown that the trend of increasing initial release as the Na2HPO4 content increased from 2 (w / v)% was due to the above-mentioned polymer aggregation phenomenon.

[0209] Experimental Example 5. Method for measuring sodium and phosphorus residues

[0210] In order to measure the content of sodium and phosphorus in the microspheres used in the embodiments and comparative examples, 6mL of nitric acid aqueous solution containing 300mg of microspheres mixed with ultrapure water in a ratio of 1: 1 was mixed with 3mL of hydrogen peroxide and then heated at 100°C or higher. Acid was added until the gas produced during dissolution changed from yellow to white. The sample thus obtained was weighed, dissolved in ultrapure water, and then injected into an inductively coupled plasma emission spectrometer (ICP-OES) (iCAP 6300Duo, Thermo Fisher Scientific, UK), and measured at a detection wavelength of 598.5nm. The results are shown in the following Experimental Examples 5-1 and 5-2.

[0211] Experimental Example 5-1. Comparison of the residual sodium and phosphorus in the microspheres according to the amount of sodium hydrogen phosphate added

[0212] The residual amounts of sodium and phosphorus in the microspheres of Preparation Examples 59 to 61 and 63 were measured and are shown in Table 13 below.

[0213] [Table 13]

[0214]

[0215]

[0216] Referring to Table 13, it was confirmed that the residual amounts of sodium and phosphorus increased significantly as the amount of Na2HPO4 added increased. The residual content of the initial release inhibitor in the microspheres was 10 ppm to 200 ppm based on sodium and 5 ppm to 100 ppm based on phosphorus.

[0217] Experimental Example 5-2. Sodium and phosphorus residues in microspheres according to biodegradable polymer changes and benzathine penicillin addition Comparison of retention

[0218] The residual amounts of Na and P in Preparation Example 16, Preparation Example 63 and Preparation Example 67 were measured and shown in Table 14.

[0219] [Table 14]

[0220]

[0221] Referring to Table 14, it was confirmed that the contents of residual sodium and residual phosphorus were different depending on the preparation conditions of the microspheres containing semaglutide and the initial release inhibitor contained in the continuous phase, and the residual content of the initial release inhibitor in the microspheres was 50 ppm to 500 ppm based on sodium and 10 ppm to 100 ppm based on phosphorus.

[0222] Experimental Example 6. Comparison of SEM results of microspheres according to whether disodium hydrogen phosphate is used in the continuous phase

[0223] Figure 3a Microspheres prepared using only 0.1 (w / v)% PVA in the continuous phase are shown. Figure 3b Shown are microspheres prepared using 0.1 (w / v)% PVA and 2 (w / v)% Na2HPO4 in the continuous phase.

[0224] Reference Figure 3a and Figure 3b As shown, it was confirmed that microparticles prepared using only 0.1% PVA in the continuous phase had many pores in the cross section of the microspheres. It was confirmed that microspheres prepared using 0.1 (w / v)% PVA and 2 (w / v)% Na2HPO4 in the continuous phase had most of the pores disappear, or even if there were pores, the number and size of the pores were significantly reduced. This can be considered to show the effect of reducing the channels that can affect the initial release of the drug.

Claims

1. A pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta cell function protection, obesity, non-alcoholic fatty hepatitis or neurodegenerative diseases, the pharmaceutical composition comprising sustained-release microspheres consisting of semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a biodegradable polymer, in, The content of semaglutide or a pharmaceutically acceptable salt thereof is 8 wt % or more based on the total weight of the microspheres as semaglutide, and the content of the bioavailability enhancer is 2.5 wt % to 250 wt % based on the weight of semaglutide.

2. A pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta cell function protection, obesity, non-alcoholic fatty hepatitis or neurodegenerative diseases, the pharmaceutical composition comprising sustained-release microspheres consisting of semaglutide or a pharmaceutically acceptable salt thereof, an initial release inhibitor and a biodegradable polymer, in, The content of semaglutide or a pharmaceutically acceptable salt thereof is 8 wt % or more based on the total weight of the microspheres in terms of semaglutide, and the content of the initial release inhibitor is 5 ppm to 2,000 ppm.

3. The pharmaceutical composition according to claim 1 or 2, wherein The sustained-release microspheres release less than 20% of semaglutide or a pharmaceutically acceptable salt thereof within 24 hours when administered in vivo.

4. The pharmaceutical composition according to claim 1 or 2, wherein The sustained-release microspheres release less than 15% of semaglutide or a pharmaceutically acceptable salt thereof within 24 hours when administered in vivo.

5. The pharmaceutical composition according to claim 1 or 2, wherein The sustained-release microspheres release less than 10% of semaglutide or a pharmaceutically acceptable salt thereof within 24 hours when administered in vivo.

6. The pharmaceutical composition according to claim 1 or 2, wherein The biodegradable polymer is at least one selected from the group consisting of polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA) which is a copolymer of lactide and glycolide, polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co-trimethylene carbonate (PLTMC), polyvinyl alcohol (PVA ... ), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoesters, polyanhydrides, polyanhydride-co-imides, polypropylene fumarate, pseudo-polyamino acids, polyalkyl cyanoacrylates, polyphosphazenes, polyphosphates, poly(butylene succinate / lactide) (PBSLA); copolymers or simple mixtures of two or more thereof; copolymers of the polymers with polyethylene glycol (PEG); and polymer-sugar conjugates in which the polymers or the copolymers are conjugated to sugars.

7. The pharmaceutical composition according to claim 1 or 2, wherein The pharmaceutically acceptable salt of semaglutide is sodium salt, acetate, benzoate, hydroxynaphthoate, naphthalene disulfonate or pamoate of semaglutide.

8. The pharmaceutical composition according to claim 1, wherein The bioavailability enhancer is at least one of the group consisting of sodium decanoate, disodium hydrogen phosphate, choline, meglumine, basic aluminum carbonate, sodium dihydroxyaluminum carbonate, ammonium phosphate, histidine, HEPES, HEPPS, spermine, spermidine, putrescine, methylene blue, proline, sugar, glycerol, surfactant, arginine, glycine, guanidine hydrochloride, urea, sodium chloride, potassium chloride, triethylamine, ethanolamine, triethanolamine, ethylenediamine, poloxamer, benzathine penicillin, procaine, lidocaine, bupivacaine, ropivacaine, oxytetracycline, sunitinib, rhizobactin, benzofuran, magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium trisilicate, zinc carbonate, zinc hydroxide, zinc phosphate, aluminum hydroxide, aluminum phosphate, dihydroxyaluminum aminoacetate, calcium phosphate, calcium hydroxide, magnesium aluminum hydroxide and benzofuran derivatives.

9. The pharmaceutical composition according to claim 1, further comprising 5 ppm to 2,000 ppm of an initial release inhibitor.

10. The pharmaceutical composition according to claim 2 or 9, wherein The initial release inhibitor is at least one substance selected from alkali metal, alkaline earth metal or ammonium phosphate salts, phosphide salts, carbonates, chromates, dichromates, oxides, oxalates, silicates, sulfates, sulfide salts, sulfites, tartrates, tetraborates, thiosulfates, arsenates, arsenites, citrates, ferrocyanide salts and nitride salts.

11. The pharmaceutical composition according to claim 1 or 2, wherein The sustained-release microspheres have an average particle size of 5 μm to 100 μm.

12. The pharmaceutical composition according to claim 1 or 2, wherein The sustained-release microspheres have a weight of 20 mg to 1,000 mg.

13. The pharmaceutical composition according to claim 1 or 2, wherein The biodegradable polymer has an intrinsic viscosity of 0.16 dL / g to 1.7 dL / g.

14. The pharmaceutical composition of claim 1 or 2, further comprising one or more release controlling agents selected from the group consisting of butyric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, beheic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, naphthalene dicarboxylic acid, naphthalenesulfonic acid and pamoic acid.

15. A method for preparing microspheres composed of semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a biodegradable polymer, the method comprising the following steps: (a) dissolving semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and one or more biodegradable polymers in an organic solvent to prepare a solution (dispersed phase) containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer; (b) adding the solution containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer prepared in step (a) to an aqueous solution phase (continuous phase) containing a surfactant to prepare an emulsion; (c) extracting an organic solvent from the dispersed phase in the emulsion state prepared in step (b) into a continuous phase, and evaporating the organic solvent to form microspheres; and (d) recovering the microspheres from the continuous phase of step (c).

16. The method according to claim 15, wherein: The bioavailability enhancer is at least one selected from the group consisting of poloxamer, benzathine penicillin, procaine, lidocaine, bupivacaine, ropivacaine, oxytetracycline, sunitinib, rhizobactin and benzofuran.

17. A method for preparing microspheres composed of semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a biodegradable polymer, the method comprising the following steps: (a') dissolving semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer in an aqueous solution to prepare a primary aqueous solution phase, dissolving one or more biodegradable polymers in an organic solvent to prepare an oil phase, and then mixing the primary aqueous solution and the oil phase to prepare a W / O emulsion (primary emulsion) containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer; (b') adding the primary emulsion containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer prepared in step (a') to an aqueous solution phase (continuous phase) containing a surfactant to prepare a W / O / W emulsion (secondary emulsion); (c') extracting an organic solvent from the oil phase in the secondary emulsion state prepared in step (b') into a continuous phase, and evaporating the organic solvent to form microspheres; and (d') recovering the microspheres from the continuous phase of step (c').

18. The method according to claim 17, wherein: The bioavailability enhancer is at least one selected from the group consisting of sodium caprate, disodium hydrogen phosphate, choline, meglumine, basic aluminum carbonate, sodium dihydroxyaluminum carbonate, ammonium phosphate, histidine, HEPES, HEPPS, spermine, spermidine, putrescine, methylene blue, proline, sugar, glycerol, surfactant, arginine, glycine, guanidine hydrochloride, urea, sodium chloride, potassium chloride, triethyleneamine, ethanolamine, triethanolamine and ethylenediamine.

19. A method for preparing microspheres composed of semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a biodegradable polymer, the method comprising the following steps: (a”) dissolving semaglutide or a pharmaceutically acceptable salt thereof and one or more biodegradable polymers in an organic solvent, and suspending a bioavailability enhancer in the solution to prepare a suspension (dispersed phase) containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer; (b") adding the suspension containing semaglutide or a pharmaceutically acceptable salt thereof, a bioavailability enhancer and a polymer prepared in step (a") to an aqueous solution phase (continuous phase) containing a surfactant to prepare an emulsion; (c") extracting an organic solvent from the dispersed phase in the emulsion state prepared in step (b") into a continuous phase, and evaporating the organic solvent to form microspheres; and (d") recovering the microspheres from the continuous phase of step (c").

20. The method according to claim 19, wherein: The bioavailability enhancer is at least one selected from the group consisting of magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium trisilicate, zinc carbonate, zinc hydroxide, zinc phosphate, aluminum hydroxide, aluminum phosphate, dihydroxyaminoacetic acid aluminum, calcium phosphate, calcium hydroxide, magnesium aluminum hydroxide and benzofuran derivatives.

21. The method according to any one of claims 15 to 20, wherein: The biodegradable polymer is at least one selected from the group consisting of polylactide (PLA), polyglycolide (PGA), polylactide-co-glycolide (PLGA) which is a copolymer of lactide and glycolide, polydioxanone, polycaprolactone (PCL), polylactide-co-glycolide-co-caprolactone (PLGC), polylactide-co-hydroxymethyl glycolide (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co-trimethylene carbonate (PLTMC), polyvinyl alcohol (PVA ... ), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoesters, polyanhydrides, polyanhydride-co-imides, polypropylene fumarate, pseudo-polyamino acids, polyalkyl cyanoacrylates, polyphosphazenes, polyphosphates, poly(butylene succinate / lactide) (PBSLA); copolymers or simple mixtures of two or more thereof; copolymers of the polymers with polyethylene glycol (PEG); and polymer-sugar conjugates in which the polymers or the copolymers are conjugated to sugars.

22. The method of any one of claims 15 to 20, further comprising one or more release controlling agents selected from the group consisting of butyric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, hydroxynaphthoic acid, naphthalene dicarboxylic acid, and pamoic acid.

23. The method according to any one of claims 15 to 20, wherein: The organic solvent in step (a) or (a') or (a") is at least one selected from the group consisting of dichloromethane, chloroform, ethyl acetate, methyl ethyl ketone, acetone, acetonitrile, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, acetic acid, methanol, ethanol, propanol and benzyl alcohol.

24. The method according to any one of claims 15 to 20, wherein: The surfactant in step (b) or (b') or (b") is polyvinyl alcohol.

25. The method according to any one of claims 15 to 20, wherein: The continuous phase in step (b) or (b') or (b") is water, or a mixed solvent of water and one or more selected from the group consisting of methanol, ethanol, propanol and ethyl acetate.

26. The method according to any one of claims 15 to 20, wherein: The continuous phase in step (b) or (b') or (b") comprises an initial release inhibitor.

27. The method according to claim 26, wherein: The content of the initial release inhibitor is 0.05 w / v % to 20 w / v % based on the total volume of the continuous phase.

28. The method according to claim 26, wherein: The initial release inhibitor is at least one selected from the group consisting of disodium hydrogen phosphate, dipotassium hydrogen phosphate, and diammonium hydrogen phosphate.

29. The method according to any one of claims 15 to 20, wherein: The continuous phase in step (b) or (b') or (b") has a pH of 7 or higher.

30. The method according to any one of claims 15 to 20, wherein: When the prepared sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer are administered in vivo, the release rate of semaglutide or a pharmaceutically acceptable salt thereof within 24 hours is less than 15%.

31. The method according to any one of claims 15 to 20, wherein: When the prepared sustained-release microspheres containing semaglutide or a pharmaceutically acceptable salt thereof and a bioavailability enhancer are administered in vivo, the release rate of semaglutide or a pharmaceutically acceptable salt thereof within 24 hours is less than 10%.

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