AU2023397760A1

AU2023397760A1 - Sustained-release microspheres including glp-1 receptor agonist or pharmaceutically acceptable salt thereof and use thereof

Info

Publication number: AU2023397760A1
Application number: AU2023397760A
Authority: AU
Inventors: Ji Hyun Bae; Seung Gu Chang; Jae Pyoung Cho; Ho Il Choi; Eun Seo Jang; Ji Hyang Lee
Original assignee: Peptron Inc
Current assignee: Peptron Inc
Priority date: 2022-12-16
Filing date: 2023-12-18
Publication date: 2025-06-26
Also published as: MX2025006996A; WO2024128882A1; JP2025539630A

Abstract

The present invention relates to a sustained-release microsphere containing a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof and a sustained-release formulation. More specifically, the present invention relates to a sustained release formulation comprising a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof, wherein the formulation includes sustained release microsphere containing the GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof, one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24 dL/g, and two or more types of high-viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60 dL/g and is free of problems associated with initial burst release and delayed release, thus enabling prolonged drug release and excellent bioavailability.

Description

[DESCRIPTION] [DESCRIPTION]

[Invention Title]

SUSTAINED-RELEASE MICROSPHERES SUSTAINED-RELEASE MICROSPHERESINCLUDING INCLUDING GLP-1 GLP-1 RECEPTOR RECEPTOR AGONIST OR AGONIST OR PHARMACEUTICALLY PHARMACEUTICALLY ACCEPTABLE ACCEPTABLE SALTSALT THEREOF THEREOF AND AND USE USE THEREOF THEREOF

[Technical Field]

[0001] Thepresent

[0001] The presentinvention invention relatesto toa sustained-release relates a sustained-releasemicrosphere microsphere including including a GLP-1 a GLP-1

receptor agonist or a pharmaceutically acceptable salt thereof and a sustained-release formulation receptor agonist or a pharmaceutically acceptable salt thereof and a sustained-release formulation

including the same, and more particularly, to a method for producing the same and a use for treating including the same, and more particularly, to a method for producing the same and a use for treating

diabetes, obesity, non-alcoholic steatohepatitis or degenerative brain diseases. diabetes, obesity, non-alcoholic steatohepatitis or degenerative brain diseases.

[0002]

[Background Art]

[0003] Glucagon-likepeptide-1

[0003] Glucagon-like peptide-1receptor receptoragonists agonists(GLP-1 (GLP-1 RA) RA) are aare a group group of drugs of drugs used for used for

treating type 2 diabetes and are very effective in lowering blood sugar levels. In addition, as a treating type 2 diabetes and are very effective in lowering blood sugar levels. In addition, as a

weight-loss effect, and an alleviation effect of hypertension, hypoglycemia and/or hyperlipidemia, weight-loss effect, and an alleviation effect of hypertension, hypoglycemia and/or hyperlipidemia,

and a cardiovascular protection effect are also known, the clinical importance thereof is increasing. and a cardiovascular protection effect are also known, the clinical importance thereof is increasing.

In addition, In addition, there there is is an advantage ofoflowering an advantage loweringa arisk riskofofcausing causing hypoglycemia hypoglycemia compared compared to to

conventional insulin secretagogues such as sulfonylurea or meglitinide, but they require frequent conventional insulin secretagogues such as sulfonylurea or meglitinide, but they require frequent

injections, which injections, reduces convenience which reduces conveniencefor forpatients patientswith withchronic chronicdiseases diseaseswho who need need long-term long-term

administration. administration.

[0004] Accordingly,there

[0004] Accordingly, thereisisa ademand demandfor for the the development development of long-acting of long-acting drugsdrugs capable capable of of

reducing the burden on patients, and as an administration cycle of drugs is increased, convenience reducing the burden on patients, and as an administration cycle of drugs is increased, convenience

for patients is increased and thus market competitiveness is further increasing. A first GLP-1 RA for patients is increased and thus market competitiveness is further increasing. A first GLP-1 RA

drug, Byetta (exenatide), was a twice-a-day injection, and then a once-a-day injection, Victoza drug, Byetta (exenatide), was a twice-a-day injection, and then a once-a-day injection, Victoza

1

(liraglutide) was (liraglutide) wasdeveloped developed and and dominated in the dominated in the market. Currently,asasonce-a-week market. Currently, once-a-weekinjections, injections,

products such as Ozempic (semaglutide) and Trulicity (dulaglutide) occupy most of the market. products such as Ozempic (semaglutide) and Trulicity (dulaglutide) occupy most of the market.

[0005] Thegreat

[0005] The greatsuccess successof of GLP-1 RA GLP-1 RA drugsincreases drugs increasesaademand demand fordrugs for drugswith withaa prolonged prolongedlong- long-

acting duration, such as once-a-month or longer. However, due to technological limitations, there acting duration, such as once-a-month or longer. However, due to technological limitations, there

is still no longer-acting drug with a longer dosage interval than once a week. is still no longer-acting drug with a longer dosage interval than once a week.

[0006] Meanwhile,

[0006] Meanwhile, sustained-release sustained-release formulations formulations using using biodegradable biodegradable polymers polymers have been have been

developedtotoachieve developed achievelong-term long-term sustained-release sustained-release effects. effects. However, However, in the in theofcase case theseof these

formulations, biodegradable polymers that release drugs more slowly are used in order to extend formulations, biodegradable polymers that release drugs more slowly are used in order to extend

the drug release duration and prevent initial burst release. As a result, little drug is released or a the drug release duration and prevent initial burst release. As a result, little drug is released or a

cumulative release rate of the drug is low at 30% or less for a considerable period of time (2 to 3 cumulative release rate of the drug is low at 30% or less for a considerable period of time (2 to 3

weeksatatthe weeks theshortest shortest and and1 1month monthor or more more at the at the longest) longest) after after administration, administration, leading leading to to a a

significant lag phase in drug release, during which a therapeutic gap may occur. significant lag phase in drug release, during which a therapeutic gap may occur.

[0007]

[Disclosure]

[Technical Problem]

[0008] Anobject

[0008] An objectofofthe thepresent presentinvention inventionisis to to provide a sustained-release provide a sustained-release microsphere having microsphere having

excellent bioavailability and a sustained-release formulation including the same. excellent bioavailability and a sustained-release formulation including the same.

[0009] Another

[0009] Another object object of present of the the present invention invention is to provide is to provide a sustained-release a sustained-release microsphere microsphere and and

a sustained-release a sustained-release formulation whichreleases formulation which releases aadrug drugcontinuously continuouslyforfora along long period period of of time time

without initial without initial burst burstrelease releaseand andhas hasno no delayed delayed release release (lag (lagphase) phase)problem, problem, and a method and a for method for

producingthe producing the same. same.

[0010] Yetanother

[0010] Yet anotherobject objectofofthe thepresent present invention invention is is to to provide provide aa pharmaceutical pharmaceuticalcomposition composition

including the including the sustained-release sustained-release microsphere or sustained-release microsphere or sustained-release formulation, formulation, and a method and a for method for

treating a disease including administering the pharmaceutical composition to a subject. treating a disease including administering the pharmaceutical composition to a subject.

2

[0011]

[Technical Solution]

[0012]

[0012] An An aspect aspect of the of the present present invention invention provides provides a sustained-release a sustained-release microsphere microsphere comprising comprising a a

GLP-1receptor GLP-1 receptoragonist agonist or or a pharmaceutically a pharmaceutically acceptable acceptable salt thereof salt thereof and poly(lactide-co- and poly(lactide-co-

glycolide), wherein the GLP-1 receptor agonist or the pharmaceutically acceptable salt thereof is glycolide), wherein the GLP-1 receptor agonist or the pharmaceutically acceptable salt thereof is

contained in an amount of 3 to 12 wt% of the total wt%, and the PLGA is a mixture of one type of contained in an amount of 3 to 12 wt% of the total wt%, and the PLGA is a mixture of one type of

low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24 dL/g and two or more types of high- low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24 dL/g and two or more types of high-

viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60 dL/g. viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60 dL/g.

[0013] Theone

[0013] The onetype typeofoflow-viscosity low-viscosityPLGA PLGAand and the the twotwo or more or more types types of high-viscosity of high-viscosity PLGA PLGA

in the mixture may have a weight ratio of 1 : 0.2 to 5 in the mixture may have a weight ratio of 1 : 0.2 to 5

[0014] TheGLP-1

[0014] The GLP-1 receptor receptor agonist agonist maymay be be semaglutide semaglutide or tirzepatide. or tirzepatide.

[0015] Thetwo

[0015] The twoorormore more types types ofof high-viscosityPLGA high-viscosity PLGAmaymay be one be one or more or more typestypes of PLGA of PLGA with with

a molar ratio of lactide to glycolide of 45 to 55 : 45 to 55 and one or more types of PLGA with a a molar ratio of lactide to glycolide of 45 to 55 : 45 to 55 and one or more types of PLGA with a

molar ratio of lactide to glycolide of 60 to 80 : 20 to 40. molar ratio of lactide to glycolide of 60 to 80 : 20 to 40.

[0016] Thepoly(lactide-co-glycolide)

[0016] The poly(lactide-co-glycolide)may maybe be included included in in an an amount amount ofto8897towt% of 88 97 of wt% theof the

sustained-release microsphere. sustained-release The sustained-release microsphere. The sustained-release microsphere microsphere may be prepared may be prepared byby by

dissolving the dissolving the GLP-1 receptoragonist GLP-1 receptor agonistororthe thepharmaceutically pharmaceuticallyacceptable acceptable saltthereof salt thereofand andthethe

poly(lactide-co-glycolide) in glacial acetic acid, spraying the solution using ultrasonic spray poly(lactide-co-glycolide) in glacial acetic acid, spraying the solution using ultrasonic spray

nozzles, and then evaporating the solvent using dry air. nozzles, and then evaporating the solvent using dry air.

[0017] Thefrequency

[0017] The frequency of of thethe ultrasonicspray ultrasonic spraynozzles nozzles maymay be to be 40 4080to kHz. 80 kHz. The sustained- The sustained-

release microsphere release maybebeadministered microsphere may administered to to S.D S.D ratsororminipigs rats minipigs andand then then thethe GLP-1 GLP-1 receptor receptor

agonist or the pharmaceutically acceptable salt thereof may be released at less than 5% within 24 agonist or the pharmaceutically acceptable salt thereof may be released at less than 5% within 24

hours, and hours, released at and released at 10% or more 10% or morewithin within11week, week,20% 20%or or more more within within 9 days, 9 days, or or 50% 50% or more or more

within 22 weeks. within weeks.

3

[0018]

[0018] TheThe sustained-release sustained-release microsphere microsphere may be may be administered administered at intervals at intervals of one to of onemonths. three to three months.

[0019] Thesustained-release

[0019] The sustained-releasemicrosphere microspheremay may have have a mean a mean diameter diameter of of 15 15 to to µm.m. 25 25 um.

[0020] Another

[0020] Another aspect aspect of present of the the present invention invention provides provides a sustained-release a sustained-release formulation formulation including including

the sustained-release microsphere according to the present invention. the sustained-release microsphere according to the present invention.

[0021] Yetanother

[0021] Yet anotheraspect aspectofofthe thepresent presentinvention inventionprovides providesa apharmaceutical pharmaceutical composition composition forfor

treating or preventing diabetes, obesity, non-alcoholic steatohepatitis or degenerative brain disease, treating or preventing diabetes, obesity, non-alcoholic steatohepatitis or degenerative brain disease,

including the sustained-release microsphere or the sustained-release formulation. including the sustained-release microsphere or the sustained-release formulation.

[0022] Thedegenerative

[0022] The degenerative braindisease brain diseasemaymay be any be any one one selected selected fromfrom the group the group consisting consisting of of

Parkinson’s disease, Parkinson's disease, Alzheimer's Alzheimer’s disease, disease, Huntington's Huntington’s disease, disease, Amyotrophic Amyotrophiclateral lateral

sclerosis(ALS), Creutzfeldt-Jakob disease, stroke, and multiple sclerosis. sclerosis(ALS), Creutzfeldt-Jakob disease, stroke, and multiple sclerosis.

[0023] Stillanother

[0023] Still another aspect aspect of the of the present present invention invention provides provides a method a method for producing for producing a sustained- a sustained-

release microsphere release microsphereincluding: including:dissolving dissolving3 to 3 to 12 12 wt% wt% of a receptor of a GLP-1 GLP-1 receptor agonist agonist or a or a

pharmaceutically acceptable pharmaceutically acceptablesalt salt thereof thereof and and8888toto9797wt%wt% of poly(lactide-co-glycolide) of poly(lactide-co-glycolide) in in a a

solvent to prepare a mixed solution; and spraying the mixed solution using ultrasonic spray nozzles, solvent to prepare a mixed solution; and spraying the mixed solution using ultrasonic spray nozzles,

and then evaporating the solvent using dry air to prepare microspheres, wherein the poly(lactide- and then evaporating the solvent using dry air to prepare microspheres, wherein the poly (lactide- poly(lactide-

co-glycolide) is a mixture of one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to co-glycolide) is a mixture of one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to

0.24 dL/g and two or more types of high-viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60 0.24 dL/g and two or more types of high-viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60

dL/g. dL/g.

[0024] Thesolvent

[0024] The solventmaymay be be an organic an organic solvent, solvent, preferably preferably a glacial a glacial acetic acetic acid acid or or aceticacid acetic acid

solution, but is not limited thereto. solution, but is not limited thereto.

[0025] Stillanother

[0025] Still another aspect aspect of the of the present present invention invention provides provides a method a method for producing for producing a sustained- a sustained-

release formulation release bydispersing formulation by dispersingthe theprepared preparedsustained-release sustained-releasemicrospheres microspheres in in an an aqueous aqueous

solution containing polyvinyl alcohol and lysine hydrochloride, and then filtering and washing, solution containing polyvinyl alcohol and lysine hydrochloride, and then filtering and washing,

and then recovering the prepared sustained-release microspheres. and then recovering the prepared sustained-release microspheres.

4

[0026] Themethod

[0026] The method for producing for producing the sustained-release the sustained-release formulation formulation mayinclude may further further include

suspending the recovered sustained-release microspheres in a solution containing D-mannitol and suspending the recovered sustained-release microspheres in a solution containing D-mannitol and

lysine hydrochloride, followed by freeze-drying. lysine hydrochloride, followed by freeze-drying.

[0027]

[Advantageous Advantageous Effects] Effects]

[Advantageous Effects]

[0028] According

[0028] According to present to the the present invention, invention, the sustained-release the sustained-release microsphere microsphere and the sustained- and the sustained-

release formulation including the same exhibit no initial burst release and enable prolonged drug release formulation including the same exhibit no initial burst release and enable prolonged drug

release. In addition, the sustained-release microsphere and the sustained-release formulation have release. In addition, the sustained-release microsphere and the sustained-release formulation have

no delayed release problem and excellent bioavailability, and can be administered to subjects at no delayed release problem and excellent bioavailability, and can be administered to subjects at

intervals of one month or more, thereby improving patient dosing convenience. intervals of one month or more, thereby improving patient dosing convenience.

[0029]

[Description of Drawings

[Description of Drawings] Drawings]

[0030] FIG.1 1isisaagraph

[0030] FIG. graphshowing showingthethe changes changes in in blood blood concentration concentration of of drugs drugs over over timetime after after

subcutaneousinjection subcutaneous injection of of sustained-release sustained-releaseformulations formulations containing containing microspheres microspheres of of Examples Examples 11

to 10 to 10 (FIGS. (FIGS. 1A, 1B, and 1A, 1B, and 1C) 1C)and andComparative Comparative Examples Examples 1 to 1 to 4 (FIG. 4 (FIG. 1D)1D) into into S.D. S.D. rats. rats.

[0031] FIG.

[0031] FIG. 2 aisgraph 2 is a graph showing showing the changes the changes in cumulative in cumulative release release rate of a rate drug of a drug over over time after time after

subcutaneous injection of sustained-release formulations containing the microspheres of Example subcutaneous injection of sustained-release formulations containing the microspheres of Example

3 and Comparative Examples 1 to 4 into S.D. rats. 3 and Comparative Examples 1 to 4 into S.D. rats.

[0032] FIG.3 3isisaa graph

[0032] FIG. graphshowing showingthethechanges changes in in blood blood concentration concentration of of a drug a drug over over time time after after

subcutaneous injection of a sustained-release formulation containing the microspheres of Example subcutaneous injection of a sustained-release formulation containing the microspheres of Example

3 into minipigs. 3 into minipigs.

[0033] FIG.

[0033] FIG. 4 aisgraph 4 is a graph showing showing the changes the changes in cumulative in cumulative release release rate of a rate drug of a drug over over time after time after

3 into minipigs. 3 into minipigs.

5

[0034] FIG.5 5isis aa graph

[0034] FIG. graph showing the body showing the weightmeasurements body weight measurements (FIG. (FIG. 5A) 5A) and and body body weight weight gain gain

rates (FIG. rates (FIG. 5B) over time 5B) over time during during88weeks weeksof of administrationofofa asustained-release administration sustained-releaseformulation formulation

containing the containing the microspheres of Example microspheres of Example 33to to aa DIO DIOmouse mouse model. model.

[0035] FIG.6 6isisa agraph

[0035] FIG. graph showing showing the the changes changes in food in food intake intake over during over time time during 8 weeks8 of weeks of

administration of a sustained-release formulation containing the microspheres of Example 3 to a administration of a sustained-release formulation containing the microspheres of Example 3 to a

DIO mouse DIO mousemodel. model.

[0036]

[Modes]

[0037] Thepresent

[0037] The presentinvention inventionprovides providesa asustained-release sustained-releasemicrosphere microsphereandand a sustained-release a sustained-release

formulation including a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof. formulation including a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof.

[0038] Thepresent

[0038] The present invention invention provides provides a sustained-release a sustained-release formulation formulation including including a GLP-1 a GLP-1

receptor agonist or a pharmaceutically acceptable salt thereof, in which the formulation includes a receptor agonist or a pharmaceutically acceptable salt thereof, in which the formulation includes a

sustained-release microsphere sustained-release microspherecontaining containinga GLP-1 a GLP-1 receptor receptor agonist agonist or a pharmaceutically or a pharmaceutically

acceptable salt thereof, one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24 acceptable salt thereof, one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24

dL/g, and dL/g, and two twooror more moretypes typesofofhigh-viscosity high-viscosityPLGA PLGA with with an an intrinsicviscosity intrinsic viscosityofof0.32 0.32toto0.60 0.60

dL/g and is free of problems associated with initial burst release and delayed release, thus enabling dL/g and is free of problems associated with initial burst release and delayed release, thus enabling

prolonged drug release and excellent bioavailability. prolonged drug release and excellent bioavailability.

[0039] Thepresent

[0039] The presentinvention inventionprovides providesa asustained-release sustained-releasemicrosphere microsphereandand a sustained-release a sustained-release

formulation comprising a sustained-release microsphere containing a GLP-1 receptor agonist or a formulation comprising a sustained-release microsphere containing a GLP-1 receptor agonist or a

pharmaceutically acceptable pharmaceutically acceptablesalt salt thereof thereofand andpoly(lactide-co-glycolide), poly(lactide-co-glycolide), ininwhich whichthethe GLP-1 GLP-1

receptor agonist or the pharmaceutically acceptable salt thereof is contained in an amount of 3 to receptor agonist or the pharmaceutically acceptable salt thereof is contained in an amount of 3 to

12 12 wt% 12 wt% wt% ofof of thethe sustained-release sustained-release the microsphere, microsphere, sustained-release and and theand microsphere, thepoly(lactide-co-glycolide) poly(lactide-co-glycolide) poly(lactide-co-glycolide) the is a mixtureis isofaa mixture mixture of of

one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24 dL/g and two or more one type of low-viscosity PLGA with an intrinsic viscosity of 0.14 to 0.24 dL/g and two or more

types of high-viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60 dL/g. types of high-viscosity PLGA with an intrinsic viscosity of 0.32 to 0.60 dL/g.

6

[0040] TheGLP-1

[0040] The GLP-1 receptor receptor agonist agonist referstotoa aglucagon-like refers glucagon-likepeptide-1 peptide-1receptor receptor agonist agonist (GLP-1 (GLP-1

RA). RA).

[0041] TheGLP-1

[0041] The GLP-1 receptor receptor agonist agonist mayanybeone may be any one selected selected from from the theconsisting group group consisting of of

exenatide, lixisenatide, albiglutide, dulaglutide, liraglutide, semaglutide, tirzepatide, retatlutide, exenatide, lixisenatide, albiglutide, dulaglutide, liraglutide, semaglutide, tirzepatide, retatlutide,

cagrilintide, cotadutide, mazdutide, and cagrisema. cagrilintide, cotadutide, mazdutide, and cagrisema.

[0042] Inone

[0042] In oneembodiment, embodiment, theGLP-1 the GLP-1 receptor receptor agonist agonist is issemaglutide. semaglutide.Semaglutide Semaglutide (NN9536) (NN9536)

is an analogue of human glucagon-like peptide-1, GLP-1(7-37), with two amino acid substitutions is an analogue of human glucagon-like peptide-1, GLP-1(7-37), with two amino acid substitutions

(Ala8 is substituted (Ala8 is substituted to to Aib8 [2-aminobutyricacid] Aib8 [2-aminobutyric acid]and andLys34 Lys34 is is substitutedtotoArg34). substituted Arg34). The The

chemical name chemical nameofofsemaglutide N6,26-{18-[N-(17carboxyheptadecanoyl)-L--glutamyl]-10-oxo- semaglutideisisN6,26-{18-[N-(17carboxyheptadecanoyl)-L-y-glutamy1]-10-oxo- N62²6-{18-[N-(17carboxyheptadecanoyl)-L-y-glutamyl]-10-oxo-

3,6,12,15-tetraoxa-9,18-diazaoctanoyl}-[8(2-amino-2-propanoic 3,6,12,15-tetraoxa-9,18-diazaoctanoy1}-[8(2-amino-2-propanoic acid),34-L-arginine] 3,6,12,15-tetraoxa-9,18-diazaoctanoyl}-[8(2-amino-2-ppopanoic acid), acid), 34-L-arginine] 34-L-arginine] human human human

glucagon-like peptide glucagon-like peptide 1(7-37). Thesemaglutide 1(7-37). The semaglutide maymay be prepared be prepared as described as described in Example in Example 4 of4 of

WO2006/097537. WO 2006/097537.

[0043] Inone

[0043] In oneembodiment, embodiment, the GLP-1 the GLP-1 receptor receptor agonistagonist is tirzepatide. is tirzepatide. The tirzepatide The tirzepatide is is

described in described in Example Example 11 of of U.S. U.S. Patent Patent No. 9,474,780. No. 9,474,780.

[0044]

[0044] TheThe “pharmaceutically "pharmaceutically acceptable acceptable salt" of salt” of the invention the present present invention refers refers to any salttowhich, any salt which,

at a concentration that is relatively non-toxic and harmless to a patient and has an effective action, at a concentration that is relatively non-toxic and harmless to a patient and has an effective action,

does not reduce the beneficial efficacy of the GLP-1 receptor agonist due to side effects caused by does not reduce the beneficial efficacy of the GLP-1 receptor agonist due to side effects caused by

the salt, and may be acid addition salts or base addition salts. the salt, and may be acid addition salts or base addition salts.

[0045] Thepharmaceutically

[0045] The pharmaceutically acceptable acceptable salt salt of of thethe GLP-1 GLP-1 receptor receptor agonist agonist may may be be an acid an acid

addition salt addition salt of ofthe theGLP-1 receptor agonist. GLP-1 receptor Forexample, agonist. For example,thetheacid acidaddition additionsalt salt of of the the GLP-1 GLP-1

receptor agonist receptor agonist may mayinclude include a hydrochloride, a hydrochloride, hydrobromide, hydrobromide, hydroiodide, hydroiodide, nitrate, nitrate, sulfate, sulfate,

bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate,

pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate,

saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p- saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-

7 toluenesulfonate, butyrate, toluenesulfonate, butyrate, camphorate, camphorate, camphorsulfonate, camphorsulfonate, digluconate, digluconate, glycerophosphate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, 2-hydroxyethanesulfonate (isethionate), nicotinate, hemisulfate, heptanoate, hexanoate, fumarate, 2-hydroxyethanesulfonate (isethionate), nicotinate,

2-naphthalenesulfonate, oxalate, 2-naphthalenesulfonate, oxalate, pectinate, pectinate, persulfate, persulfate, 3-phenylpropionate, 3-phenylpropionate,picrate, picrate,pivalate, pivalate,

propionate, thiocyanate, or bicarbonate. propionate, thiocyanate, or bicarbonate.

[0046]

[0046] TheThe pharmaceutically pharmaceutically acceptable acceptable salt ofsalt the of the receptor GLP-1 GLP-1 receptor agonist agonist may mayaddition be a base be a base addition

salt of salt of the theGLP-1 receptor agonist. GLP-1 receptor agonist. For Forexample, example, thebase the baseaddition additionsalt saltofofthe the GLP-1 GLP-1 receptor receptor

agonist may be an alkali metal salt, an alkaline earth metal salt or a quaternary ammonium salt. agonist may be an alkali metal salt, an alkaline earth metal salt or a quaternary ammonium salt.

[0047] Thesustained-release

[0047] The sustained-releasemicrosphere microsphere of the of the present present invention invention includes includes polylactide-co- polylactide-co-

glycolide (PLGA) glycolide togetherwith (PLGA) together withthe theGLP-1 GLP-1 receptor receptor agonist agonist or or thethe pharmaceutically pharmaceutically acceptable acceptable

salt thereof. salt thereof.

[0048] Thesustained-release

[0048] The sustained-releasemicrosphere microsphereincludes includesatatleast least one type of one type of low-viscosity low-viscosity PLGA and PLGA and

at least two types of high-viscosity PLGA. at least two types of high-viscosity PLGA.

[0049] Asused

[0049] As usedininthe the present present invention, invention,the the“low-viscosity PLGA” "low-viscosity PLGA" means PLGA means PLGA with with an an intrinsic intrinsic

viscosity of 0.14 to 0.24 dL/g. viscosity of 0.14 to 0.24 dL/g.

[0050] The"high-viscosity

[0050] The “high-viscosityPLGA" PLGA” means means PLGAPLGA with with an an intrinsic intrinsic viscosity viscosity of of 0.32 0.32 to to 0.60dL/g. 0.60 dL/g.

At least two types of high-viscosity PLGA are included. At least two types of high-viscosity PLGA are included.

[0051] Thelow-viscosity

[0051] The low-viscosityPGLA PGLAand and the the high-viscosity high-viscosity PLGA PLGA may may be be included included in a in a weight weight ratioratio

of 1 : 0.2 to 5, 1 : 0.2 to 3, 1 : 0.3 to 5, 1 : 0.3 to 3, 1 : 0.5 to 5, 1: 0.5 to 3, 1 : 0.5 to 2, 1 : 1 to 5, of 1 : 0.2 to 5, 1 1:: 0.2 0.2 to to 3, 3, 11 :: 0.3 0.3 to to 5, 5, 11 :: 0.3 0.3 to to 3, 3, 11 :: 0.5 0.5 to to 5, 5, 1: 1: 0.5 0.5 to to 3, 3, 1: 1 : 0.5 0.5 toto 2,2, 1 1 : : 1 1 toto 5,5,

11 :: 11 toto 3,3,11:1:1:1to 1toto 2.5, 2.5, 2.5, 1 : 1: 11.5 1.5: to1.5 to toor2.5 2.5 2.5 or : or 1 1 : 1to:to2.2. 1.8 1.8 1.8 2.2.to 2.2.

[0052] Inone

[0052] In oneembodiment, embodiment, a mixture a mixture of of one one typeofofPLGA type PLGAwithwith a viscosity a viscosity of of 0.32toto0.44 0.32 0.44dL/g dL/g

(high-viscosity PLGA-1) (high-viscosity and PLGA-1) and oneone type type of of PLGA PLGA with with a a viscosity viscosity of to of 0.45 0.45 to dL/g 0.60 0.60 (high- dL/g (high-

viscosity PLGA-2) viscosity may PLGA-2) may bebe used used asas thehigh-viscosity the high-viscosityPLGA. PLGA.

[0053] Theweight

[0053] The weight of of thesustained-release the sustained-releasemicrosphere microsphere may may be mg be 20 20 to mg6,000 to 6,000 mg,mg20 mg, 20 tomg to

4,000 mg, 4,000 mg,100 100mgmgtoto6,000 6,000mg, mg, 100 100 mg mg to 4,000 to 4,000 mg,mg, 500 500 mg6,000 mg to to 6,000 mg, mg mg, 500 500tomg to 4,000 4,000 mg, mg,

8

1,000 1,000 mg to 6,000 mg to mg, 1,000 6,000 mg, 1,000 mg mgtoto 4,000 4,000 mg, mg,1,500 1,500mg mgtoto6,000 6,000mg, mg,1,500 1,500mgmgtoto4,000 4,000mg, mg,2,000 2,000

mgtoto 6,000 mg 6,000 mg, mg,2,000 2,000mg mgtoto4,000 4,000mg, mg,2,400 2,400mgmgtoto6,000 6,000mg, mg,2,400 2,400mgmg to to 4,000mgmg 4,000 or or 2,400 2,400 mgmg

to 3,600 to 3,600 mg. mg.

[0054] TheGLP-1

[0054] The GLP-1 receptor receptor agonist agonist or pharmaceutically or the the pharmaceutically acceptable acceptable salt thereof salt thereof may bemay be

contained in an amount of 3 to 12 wt% of the sustained-release microsphere. contained in an amount of 3 to 12 wt% of the sustained-release microsphere.

[0055] Inone

[0055] In oneembodiment, embodiment,thethe GLP-1 GLP-1 receptor receptor agonist agonist or the or the pharmaceutically pharmaceutically acceptable acceptable saltsalt

thereof may thereof be included may be included in in an an amount amountofof33wt% wt%totoless lessthan than12 12wt%, wt%,3 3wt% wt%to to 11.8 11.8 wt%, wt%, 3 wt% 3 wt%

to 10.8 to 10.8 wt%, wt%, 33 wt% wt%toto10.5 10.5wt%, wt%,3 3wt%wt% to to 10.2 10.2 wt%, wt%, 3 wt% 3 wt% to 10towt%, 10 wt%, or 3 or wt%3 to wt% to 9 or 9 wt%, wt%, or

maybebeincluded may includedinin an an amount amountofof8 8wt% wt%to to lessthan less than1212wt%, wt%,8 8 wt% wt% to to 11.8 11.8 wt%, wt%, 8 wt% 8 wt% to 10.8 to 10.8

wt%,88wt% wt%, wt%toto10.5 10.5wt%, wt%,8 8wt% wt%to to 10.2wt%, 10.2 wt%, 8 wt% 8 wt% to 10 to 10 wt%, wt%, or 8orwt% 8 wt% to 9towt%. 9 wt%. Preferably, Preferably,

the GLP-1 receptor agonist or the pharmaceutically acceptable salt thereof may be included in an the GLP-1 receptor agonist or the pharmaceutically acceptable salt thereof may be included in an

amountofof55 wt% amount wt%totoless less than than 12 12 wt%, 5 wt% wt%, 5 to 11.8 wt% to 11.8 wt%, wt%, 55 wt% wt%toto10.8 10.8 wt%, wt%,55wt% wt%toto10.5 10.5wt%, wt%,

5 wt% to 10.2 wt% to 10.2 wt%, wt%,55 wt% wt%toto1010wt%, wt%,oror5 5wt% wt%toto 9 9wt%. wt%. However, However, the present the present invention invention is not is not

limited thereto. limited thereto.

[0056] Thesustained-release

[0056] The sustained-releasemicrosphere microsphereororthe thesustained-release sustained-release formulation formulationaccording accordingtotothe the

present invention has excellent bioavailability and may be administered at intervals of one month present invention has excellent bioavailability and may be administered at intervals of one month

or longer when the content of the GLP-1 receptor agonist or the pharmaceutically acceptable salt or or longer longer when when the the content content of of the the GLP-1 GLP-1 receptor receptor agonist agonist or or the the pharmaceutically pharmaceutically acceptable acceptable salt salt

thereof is 3 to 12 wt%. In terms of initial release inhibition, it is preferred that the GLP-1 receptor thereof is 3 to 12 wt%. In terms of initial release inhibition, it is preferred that the GLP-1 receptor

agonist or the pharmaceutically acceptable salt thereof may be included in an amount of 3 to 10 agonist or the pharmaceutically acceptable salt thereof may be included in an amount of 3 to 10

wt% in the sustained-release microsphere. In terms of bioavailability, it is preferred that the GLP- wt% in the sustained-release microsphere. In terms of bioavailability, it is preferred that the GLP-

11 receptor receptoragonist agonistororthethepharmaceutically pharmaceutically acceptable acceptable salt thereof salt thereof may be in may be included included in an amount an amount

of 5 to 12 wt% or 7 to 12 wt% in the sustained-release microsphere. of 5 to 12 wt% or 7 to 12 wt% in the sustained-release microsphere.

[0057] When

[0057] When thethe twotwo or or more more types types of high-viscosity of high-viscosity PLGA PLGA are included are not not included in sustained- in the the sustained-

release microsphere, release microsphere, the the initial initialrelease amount release amountofof the the GLP-1 receptor agonist GLP-1 receptor agonist or or the the

9 pharmaceutically acceptable pharmaceutically acceptable salt salt may be excessively may be excessively increased. increased. When When the the low-viscosity low-viscosity PLGA PLGA is not included, the initial release amount of the GLP-1 receptor agonist or the pharmaceutically is not included, the initial release amount of the GLP-1 receptor agonist or the pharmaceutically acceptable salt may be small, but the delayed release (lag phase) may be increased and the total acceptable salt may be small, but the delayed release (lag phase) may be increased and the total drug release amount may be small, which may lower the bioavailability. drug drug release releaseamount maymay amount be small, whichwhich be small, may lower may the bioavailability. lower the bioavailability.

[0058] Thesustained-release

[0058] The sustained-releasemicrosphere microsphereof of thethe presentinvention present invention includes includes a poly(lactide-co- a poly(lactide-co-

glycolide) mixture glycolide) mixture in in which low-viscosity PLGA which low-viscosity and PLGA and two two or or more more types types of of high-viscosityPLGA high-viscosity PLGA

are mixed, are mixed, preferably preferably the the low-viscosity low-viscosityPLGA andthe PLGA and the two twoor or more moretypes typesof of high-viscosity high-viscosity PLGA PLGA

are mixed in a weight ratio of 1 : 0.2 to 5, more preferably the low-viscosity PLGA and the two or are mixed in a weight ratio of 1 : 0.2 to 5, more preferably the low-viscosity PLGA and the two or

moretypes more typesofof high-viscosity high-viscosity PLGA PLGA areare mixed mixed inweight in a a weight ratio ratio of of 1 :1 1.8 : 1.8toto2.2. 2.2.As As a result, a result,

while the initial burst release of the GLP-1 receptor agonist does not occur, there is no problem of while the initial burst release of the GLP-1 receptor agonist does not occur, there is no problem of

delayed release (lag phase), long-term sustained release is enabled, and excellent bioavailability is delayed release (lag phase), long-term sustained release is enabled, and excellent bioavailability is

exhibited. exhibited.

[0059] Theintrinsic

[0059] The intrinsic viscosity viscosity of of PLGA PLGA isis aa viscosity viscosity measured usingan measured using anUbbelohde Ubbelohde viscometer viscometer

at 25C at after dissolving 25°C after dissolvingPLGA in chloroform PLGA in chloroformat at aa concentration concentration of of 0.1% 0.1% (W/V). (W/V).

[0060] Low-viscosity

[0060] Low-viscosity PLGA PLGA may havemay haveratio a molar a molar ratio of of lactide to lactide toof glycolide glycolide 40 to 80 of 40toto60. : 20 80 : 20 to 60.

In one embodiment, the molar ratio of lactide to glycolide is, for example, 45 to 70 : 30 to 55, 55 In one embodiment, the molar ratio of lactide to glycolide is, for example, 45 to 70 : 30 to 55, 55

to 70 : 30 to 45 or 45 to 55 : 45 to 55. to 70 30 to to : 30 45 45 or or 45 45 to to 55 55 : 45 to to : 45 55. 55.

[0061] Low-viscosity PLGA

[0061] Low-viscosity commercialproducts PLGA commercial productsinclude include RG502, RG502,RG502H, RG502H, RG752H, RG752H, and and

RG752S RG752S from from Evonik. Evonik. In present In the the present invention, invention, RG502H RG502H is also is also referred referred to as to as PLGA50A. PLGA50A.

[0062] High-viscosity

[0062] High-viscosity PLGA PLGA may may have haveratio a molar a molar ratio oftolactide of lactide to glycolide glycolide of 40 to 80of : 40 to 80 20 to 60. : 20 to 60.

In one embodiment, the molar ratio of lactide to glycolide is, for example, 40 to 75 : 25 to 60, 45 In one embodiment, the molar ratio of lactide to glycolide is, for example, 40 to 75 : 25 to 60, 45

to 70 : 30 to 55, 55 to 70 : 30 to 45 or 45 to 55 : 45 to 55. In one embodiment, a mixture of one to 70 : 30 to 55, 55 to 70 : 30 to 45 or 45 to 55 : 45 to 55. In one embodiment, a mixture of one

type of PLGA having a molar ratio of lactide to glycolide of 45 to 55 : 45 to 55 and one or more type of PLGA having a molar ratio of lactide to glycolide of 45 to 55 : 45 to 55 and one or more

types of PLGA with a molar ratio of lactide to glycolide of 60 to 80 : 20 to 40 may be used as the types of PLGA with a molar ratio of lactide to glycolide of 60 to 80 : 20 to 40 may be used as the

10 high-viscosity PLGA. high-viscosity PLGA. InInone oneembodiment, embodiment,a amixture mixtureofofhigh-viscosity high-viscosity PLGA-1 and high- PLGA-1 and high- viscosity PLGA-2 viscosity may PLGA-2 may be be used used as as thehigh-viscosity the high-viscosityPLGA. PLGA.

[0063] High-viscosityPLGA

[0063] High-viscosity PLGA commercial commercial products products include include RG503, RG503, RG503H, RG503H, RG653H, RG653H, RG753H, RG753H,

RG753S,andand RG753S, RG504H RG504H from from Evonik. Evonik. In the present In the present invention, invention, RG503H RG503H is also referred is also referred to as to as

PLGA50B, PLGA50B, RG504H RG504H is also is also referred referred to PLGA50C, to as as PLGA50C, RG653HRG653H is also referred is also referred to as PLGA65B, to as PLGA65B,

and RG753H and RG753H is is alsoreferred also referredto to as as PLGA75B. PLGA75B.

[0064] Poly(lactide-co-glycolide) may

[0064] Poly(lactide-co-glycolide) maybebeincluded includedininananamount amount of to of 88 88 97 to wt% 97 wt% of total of the the total

weight of the sustained-release microsphere. weight of the sustained-release microsphere.

[0065]

[0065] TheThe sustained-release sustained-release formulation formulation according according to the invention to the present present invention may furthermay further include include

a coating material, additives, excipients, etc., in addition to the active ingredient and poly(lactide- a coating material, additives, excipients, etc., in addition to the active ingredient and oly(lactide- poly(lactide-

co-glycolide). co-glycolide).

[0066] Inone

[0066] In oneembodiment, embodiment, the the sustained-release sustained-release formulation formulation may include may include 3 to 123 wt% to 12 of wt% of

semaglutide, tirzepatide, or a pharmaceutically acceptable salt of either, 88 to 97 wt% poly(lactide- semaglutide, tirzepatide, or a pharmaceutically acceptable salt of either, 88 to 97 wt% poly(lactide-

co-glycolide), and a coating material, additives, excipients, or the like. co-glycolide), and a coating material, additives, excipients, or the like.

[0067]

[0067] TheThe sustained-release sustained-release microsphere microsphere may bewith may be coated coated with lysine, lysine, and and the the lysine lysine coated coated on the on the

sustained-release microsphere may be included in an amount of 0.01 to 5, 0.01 to 3, 0.01 to 1, 0.1 sustained-release microsphere may be included in an amount of 0.01 to 5, 0.01 to 3, 0.01 to 1, 0.1

to 1, 0.2 to 0.8, or 0.4 to 0.6 parts by weight based on 100 parts by weight of the sustained-release to 1, 0.2 to 0.8, or 0.4 to 0.6 parts by weight based on 100 parts by weight of the sustained-release

microspherebefore microsphere beforecoating. coating.

[0068] Thesustained-release

[0068] The sustained-releaseformulation formulationcontaining containing thethe sustained-releasemicrosphere sustained-release microsphere of of thethe

present invention may have a lasting drug effect for one month or more with a single administration, present invention may have a lasting drug effect for one month or more with a single administration,

dependingononan an depending administration administration target. target. The sustained-release The sustained-release formulation formulation of the of the present present

invention may invention maybebeadministered administeredtotoa asubject subjectatat intervals intervals of of 1 1 month, 1.5 months, month, 1.5 months,2 2months, months,2.52.5

months, or months, or 33 months. months.

[0069] Thesustained-release

[0069] The sustained-releasemicrosphere microsphere of the of the present present invention invention or sustained-release or the the sustained-release

11 formulation containing formulation containing the the sustained-release sustained-releasemicrosphere microspheremay may be be administered administered to toSprague Sprague Dawley Dawley

Rat (S.D Rat) and then the GLP-1 receptor agonist or the pharmaceutically acceptable salt thereof Rat (S.D Rat) and then the GLP-1 receptor agonist or the pharmaceutically acceptable salt thereof

is released at 5% or less within 24 hours, and released at 10% or more within 1 week, 20% or more is released at 5% or less within 24 hours, and released at 10% or more within 1 week, 20% or more

within 99 days, within days, or or50% or more 50% or within 22 weeks. more within weeks.

[0070] Themean

[0070] The mean diameter diameter of of thethe sustained-releasemicrosphere sustained-release microsphereofofthe thepresent presentinvention inventionmay maybebe

55 to 50 m, to 50 µm, 55 to um, 40 m, to 40 µm, 55 to um, 30 m, to 30 um, 10 to µm, 10 to 50 m,1010toto 40 50 um, µm, m,1010toto3030um, 40 um, µm, m,1010toto2525um, µm, m, µm, 1515 toto

50 m, 50 um, 15to µm, 15 to 40 m,15 40 um, µm, 15to to 30 moror15 30 um µm 15to to 25 m. 25 um. µm.

[0071]

[0071] TheThe spanspan value value ofsustained-release of the the sustained-release microsphere microsphere is 2 or is 2 or less, less, and moreand more specifically specifically 0.1 0.1

to 2, 0.1 to 1.8, 0.3 to 2, 0.3 to 1.8, 0.5 to 2, 0.5 to 1.8, 0.8 to 2, 0.8 to 1.8, 1 to 2, 1 to 1.8 or 1 to to 2, 0.1 to 1.8, 0.3 to 2, 0.3 to 1.8, 0.5 to 2, 0.5 to 1.8, 0.8 to 2, 0.8 to 1.8, 1 to 2, 1 to 1.8 or 1 to

1.6. 1.6.

[0072] Thesustained-release

[0072] The sustained-releasemicrosphere microsphere or or thethe sustained-releaseformulation sustained-release formulation containing containing thethe

sustained-release microsphere may further include additives and excipients commonly used in the sustained-release microsphere may further include additives and excipients commonly used in the

formulation of drugs in the art to which the present invention pertains. formulation of drugs in the art to which the present invention pertains.

[0073] Thesustained-release

[0073] The sustained-releaseformulation formulationofofthe thepresent presentinvention inventionmay may further further include include oneone or or

moreprotective more protectivecolloids colloidsselected selectedfrom fromthethe group group consisting consisting of polyvinyl of polyvinyl alcohol, alcohol, albumin, albumin,

polyvinylpyrrolidone, gelatin, polyvinylpyrrolidone, gelatin, and and the the like. like. Protective Protectivecolloidal colloidalmaterials materialsserve serve to to prevent prevent

aggregation of microspheres containing the GLP-1 receptor agonist and improve the dispersibility. aggregation of microspheres containing the GLP-1 receptor agonist and improve the dispersibility.

It is preferable that the protective colloids are included in an amount of 0.02 to 1.0 wt% with It is preferable that the protective colloids are included in an amount of 0.02 to 1.0 wt% with

respect to the total weight of the sustained-release formulation. respect to the total weight of the sustained-release formulation.

[0074] Thepresent

[0074] The presentinvention inventionprovides providesa apharmaceutical pharmaceutical composition composition forfor treatingororpreventing treating preventing

diabetes, obesity, non-alcoholic steatohepatitis (NASH) or degenerative brain disease, comprising diabetes, obesity, non-alcoholic steatohepatitis (NASH) or degenerative brain disease, comprising

the sustained-release microsphere or the sustained-release formulation containing the same. the sustained-release microsphere or the sustained-release formulation containing the same.

[0075] Thedegenerative

[0075] The degenerative braindisease brain diseasemaymay be any be any one one selected selected fromfrom the group the group consisting consisting of of

12 sclerosis(ALS), Creutzfeldt-Jakob disease, stroke, and multiple sclerosis. sclerosis(ALS), Creutzfeldt-Jakob disease, stroke, and multiple sclerosis.

[0076] Thesustained-release

[0076] The sustained-releasemicrosphere microspheremaymay be prepared be prepared by dissolving by dissolving the GLP-1 the GLP-1 receptor receptor

agonist or the pharmaceutically acceptable salt thereof and poly(lactide-co-glycolide) in glacial agonist or the pharmaceutically acceptable salt thereof and poly(lactide-co-glycolide) in glacial

acetic acid, spraying the solution using ultrasonic spray nozzles, and then evaporating the solvent acetic acid, spraying the solution using ultrasonic spray nozzles, and then evaporating the solvent

using dry air. using dry air.

[0077] Thepresent

[0077] The presentinvention inventionprovides providesa amethod methodforfor producing producing a sustained-releasemicrosphere a sustained-release microsphere

which has no initial burst release of drugs, enables long-term sustained release, has no delayed which has no initial burst release of drugs, enables long-term sustained release, has no delayed

release (lag phase) problem, and has excellent bioavailability. release (lag phase) problem, and has excellent bioavailability.

[0078] Themethod

[0078] The method for for producing producing the the sustained-release sustained-release microsphere microsphere of present of the the present invention invention

includes dissolving includes dissolving a a GLP-1 receptoragonist GLP-1 receptor agonistororaa pharmaceutically pharmaceuticallyacceptable acceptablesalt salt thereof thereof and and

poly(lactide-co-glycolide) in an organic solvent to prepare a mixed solution; and drying the mixed poly(lactide-co-glycolide) in an organic solvent to prepare a mixed solution; and drying the mixed

solution to obtain sustained-release microspheres. solution to obtain sustained-release microspheres.

[0079]

[0079] In In thethe method method for producing for producing the sustained-release the sustained-release formulation, formulation, the type ofthe type poly of poly(lactide- (lactide- poly(lactide-

co-glycolide) dissolved co-glycolide) dissolved together togetherwith withthetheGLP-1 GLP-1 receptor receptor agonist, agonist, or pharmaceutically or the the pharmaceutically

acceptable salt thereof is the same as described above for the sustained-release microsphere. acceptable salt thereof is the same as described above for the sustained-release microsphere.

[0080] Inthe

[0080] In themethod methodforforproducing producing thethe sustained-releasemicrosphere, sustained-release microsphere, thethe weight weight ratioofofthethe ratio

GLP-1 receptor agonist and poly(lactide-co-glycolide) may be appropriately adjusted so that the GLP-1 receptor agonist and poly(lactide-co-glycolide) may be appropriately adjusted SO so that the

content of content of the the GLP-1 receptoragonist GLP-1 receptor agonist in in the the produced producedsustained-release sustained-release microsphere microsphereisis33to to 12 12

wt%. wt%.

[0081] Theorganic

[0081] The organicsolvent solventmaymay be one be one or a or a mixed mixed solvent solvent thereof thereof selected selected fromgroup from the the group

consisting of chloroform, ethyl acetate, methylene chloride, methyl ethyl ketone, alcohols having consisting of chloroform, ethyl acetate, methylene chloride, methyl ethyl ketone, alcohols having

11 to to 5 5 carbon atoms,glacial carbon atoms, glacialacetic aceticacid, acid,formic formic acid, acid, dimethyl dimethyl sulfoxide, sulfoxide, and n-methylpyrrolidone, and n-methylpyrrolidone,

and preferably glacial acetic acid. and preferably glacial acetic acid.

[0082]

[0082] In In thethe method method for producing for producing the sustained-release the sustained-release formulation formulation of the of the present present ainvention, a invention,

13 spray drying spray drying method maybebeused method may usedasasthe themethod methodofofdrying dryingthe themixed mixedsolution solutiontoto obtain obtain microspheres. microspheres.

[0083] Thespray

[0083] The spraydrying dryingmethod method maymay be performed be performed by supplying by supplying a mixed a mixed solution solution in which in which the the

semaglutide oror the semaglutide the pharmaceutically pharmaceuticallyacceptable acceptablesalt saltthereof thereofand andpoly(lactide-co-glycolide) poly(lactide-co-glycolide)are are

dissolved to a spray dryer at a flow rate of 5 to 20 mL/min, 5 to 15 mL/min, or 5 to 10 mL/min, dissolved to a spray dryer at a flow rate of 5 to 20 mL/min, 5 to 15 mL/min, or 5 to 10 mL/min,

and supplying and supplyingdrying dryingair airofof100 100toto200°C, 200C,100100 to to 150C, 150°C, or 120 or 120 to 150C. to 150°C. In addition, In addition, the the

frequency of frequency of the the spray spray dryer dryer may be 40 may be 40 to to 80 80 kHz, kHz, 50 50 to to 70 70 kHz kHzoror55 55toto 75 75 kHz, kHz,and andpreferably preferably

60 kHz. 60 kHz.

[0084] Themethod

[0084] The methodforfor producing producing thethe sustained-releaseformulation sustained-release formulationofofthe the present present invention invention may may

further include further suspendingthe include suspending thedried driedmicrospheres microspheres in aqueous in an an aqueous solution solution in which in which lysine lysine

hydrochloride is dissolved to coat the microspheres with lysine, after drying the mixed solution to hydrochloride is dissolved to coat the microspheres with lysine, after drying the mixed solution to

obtain the microspheres. obtain the microspheres.

[0085] Theaqueous

[0085] The aqueous solutionininwhich solution whichlysine lysinehydrochloride hydrochlorideisisdissolved dissolvedmay mayfurther furtherinclude include1% 1%

(W/V) polyvinylalcohol. (W/V) polyvinyl alcohol.

[0086] Thepresent

[0086] The presentinvention inventionprovides provides a method a method for treating for treating diabetes, diabetes, obesity, obesity, non-alcoholic non-alcoholic

steatohepatitis or degenerative brain disease, including administering to a subject the sustained- steatohepatitis or degenerative brain disease, including administering to a subject the sustained-

release microsphere or the sustained-release formulation containing the same. release microsphere or the sustained-release formulation containing the same.

[0087] Thedegenerative

[0087] The degenerativebrain braindisease diseasemaymay be any be any one one selected selected fromfrom the group the group consisting consisting of of

sclerosis(ALS)(Lou Gehrig’s disease), Creutzfeldt-Jakob disease, stroke, and multiple sclerosis. sclerosis(ALS)(Lou Gehrig's disease), Creutzfeldt-Jakob disease, stroke, and multiple sclerosis.

[0088] Thesustained-release

[0088] The sustained-releasemicrosphere microsphere of the of the present present invention invention or sustained-release or the the sustained-release

formulation containing formulation containingthe thesustained-release sustained-releasemicrosphere microsphere may may be administered be administered viaororal via oral or

parenteral routes, parenteral routes, and preferably via and preferably viaparenteral parenteralroutes, routes,such suchas as intravenous intravenous administration, administration,

subcutaneous administration, intramuscular administration, intraperitoneal administration, etc. subcutaneous administration, intramuscular administration, intraperitoneal administration, etc.

14

[0089] Theeffective

[0089] The effectivedose dose of sustained-release of the the sustained-release microsphere microsphere or the or the sustained-release sustained-release

formulation according formulation according to to the the present present invention invention may maybebeappropriately appropriatelyadjusted adjusteddepending dependingononthethe

age of a patient, the type and degree of a disease, the condition of a patient, etc. For example, age of a patient, the type and degree of a disease, the condition of a patient, etc. For example,

the effective the effectivedose dose may be 11 to may be to 30 30 mg/week basedononananactive mg/week based activeingredient ingredientamount. amount.The The amount amount

may be administered at once or in divided doses. may be administered at once or in divided doses.

[0090] Hereinafter,

[0090] Hereinafter, the the present present invention invention will will be be described described in morein morethrough detail detail Examples. through Examples.

[0091]

[0092] Examples

[0093] 1.Preparation

[0093] 1. Preparationofofsustained-release sustained-releasemicrosphere microsphere

[0094] Semaglutide

[0094] Semaglutide oror tirzepatide, as tirzepatide, as aa GLP-1 receptoragonist, GLP-1 receptor agonist, and and PLGA PLGA as as shown shown in Table in Table 1 1

belowwere below wereprepared. prepared.

[0095] [Table1]1]

[0095] [Table

Classification Classification Poly(D,L-lactide-co-glycolide) Poly(D,L-lactide-co-glycolide) (LA (LA :: GA) Intrinsic viscosity GA) Intrinsic (dL/g) viscosity (dL/g)

Low-viscosityPLGA Low-viscosity PLGA PLGA50A (50 : PLGA50A (50 : 50) 50) 0.16 - 0.24 0.16 0.24 High-viscosity PLGA-1 High-viscosity PLGA-1 PLGA50B PLGA50B (50: : 50) (50 50) 0.32 0.32 -0.44 0.44 High-viscosity PLGA-1 High-viscosity PLGA-1 PLGA65B PLGA65B (65: : 35) (65 35) 0.32 -0.44 0.32 0.44 High-viscosity PLGA-1 High-viscosity PLGA-1 PLGA75B PLGA75B (75: : 25) (75 25) 0.32 0.32 -0.44 0.44 High-viscosity PLGA-2 High-viscosity PLGA-2 PLGA50C PLGA50C (50: : 50) (50 50) 0.45 0.45 -0.60 0.60

[0096]

[0097] Thesemaglutide

[0097] The semaglutideoror tirzepatide and tirzepatide and the the PLGA PLGA were were completely completely dissolved dissolved in in glacialacetic glacial acetic

acid to acid to obtain obtaina amixed mixed solution, solution, followed followed by drying by spray spray to drying to sustained-release prepare prepare sustained-release

microspheres. microspheres.

[0098] Thespray

[0098] The spraydrying dryingprocess processwaswas performed performed by supplying by supplying the the prepared prepared mixed mixed solution solution at a at a

flow rate flow rate of of 7.5 7.5 mL/min mL/min to to a spray a spray dryer dryer (EIN (EIN System) System) equipped equipped with with an an ultrasonic ultrasonic nozzle nozzle

(Sonictopia, 60 (Sonictopia, kHz)using 60 kHz) usinga aliquid liquidperistaltic peristaltic pump pump(Master (Master flex),adjusting flex), adjustingthe thepower power of of a a

generator (Sonictopia, generator (Sonictopia, 60 60 kHz) kHz) between 50% between 50% and and 80%, 80%, andand supplying supplying drying drying air air at at 135C. 135°C. The The

15 microspheresprepared microspheres preparedthrough throughthe thespray spraydrying dryingprocess processwere were dispersed dispersed in in an an aqueous aqueous solution solution containing 1% containing 1%(W/V) (W/V) polyvinyl polyvinyl alcohol alcohol (Mitsubishi (Mitsubishi Chemical Chemical Corporation) Corporation) and and 0.5L-lysine 0.5 M M L-lysine hydrochloride (Merck) hydrochloride (Merck) as protective as protective colloids colloids and stirred and stirred for 3 for 3 hours. hours. Thereafter, Thereafter, the the microspheres were recovered through filtration and washing with distilled water, suspended in a microspheres were recovered through filtration and washing with distilled water, suspended in a solution containing solution containing 10% D-mannitol 10% D-mannitol (ROQUETTE) (ROQUETTE) andL-lysine and 0.5% 0.5% L-lysine hydrochloride hydrochloride (Merck), (Merck), and then freeze-dried to obtain a sustained-release formulation of the present invention. and then freeze-dried to obtain a sustained-release formulation of the present invention.

[0099] Thecompositions

[0099] The compositions of of semaglutide semaglutide andand PLGAPLGA in theinsustained-release the sustained-release microsphere microsphere were were

shownasasininExamples shown Examples 1 to1 9toand 9 and Comparative Comparative Examples Examples 1 to 1 to 4 in 4 in2 below, Table Table 2andbelow, the and the

compositionsof compositions of tirzepatide tirzepatide and and PLGA were PLGA were shown shown as in as in Example Example 10 and 10 and Comparative Comparative Example Example

5. InInExample 5. Example7, 7, PLGA65B PLGA65B and PLGA75B and PLGA75B were usedwere used as high-viscosity as high-viscosity PLGA (C)PLGA in the(C) in same the same

amountasaslow-viscosity amount low-viscosity PLGA PLGA (A). (A).

[00100] [Table2]2]

[00100] [Table

Classificati Classificati PLGA PLGA GLP-1 RA GLP-1 RA Drug% Drug% on on Low- Low- High- High- High-viscosity A:B:C High-viscosity A:: BB :: CC A (w/w) (w/w) viscosity viscosity viscosity viscosity PLGA(C) PLGA(C) PLGA(A) PLGA(A) PLGA(A) PLGA(B) PLGA(B) Ex. 1 Ex. 1 PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA50C PLGA50C 11:1:1 :1:1 Semaglutide Semaglutide 8.3 8.3

Ex. 22 Ex. PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 1 11:1:1 1: 1 1 :111 Semaglutide Semaglutide 8.4 8.4

Ex. 33 Ex. PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 11:1:11 : 1 : Semaglutide Semaglutide 9.3 9.3

Ex. 44 Ex. PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 1: 1 :11 11:1:1 Semaglutide Semaglutide 10.8 10.8

Ex. 555 Ex. Ex. PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA75B PLGA75B 1: 1 :11 11:1:1 Semaglutide Semaglutide 9.3 9.3

Ex. 66 Ex. PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 11:1:2 :1:2 Semaglutide Semaglutide 9.2 9.2

Ex. 77 Ex. PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA65BPLG PLGA65BPLG 1 :1:2 1:1:2 Semaglutide Semaglutide 9.3 9.3

A75B A75B Ex. 88 Ex. PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 11:2:2 1 ::2:2 2:2 Semaglutide Semaglutide 9.3 9.3

Ex. 99 Ex. PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 11:4:1 1: 4 4 : 11 Semaglutide Semaglutide 9.6 9.6 9.6 Ex. 10 Ex. 10 PLGA50A PLGA50A PLGA50A PLGA50B PLGA50B PLGA65B PLGA65B 11:1 :11 : 1:1:1 Tirzepatide Tirzepatide Tirzepatide 11.3 11.3

Com.Ex. Com. Ex.11 PLGA50A PLGA50A PLGA50A -- - - 11:0:0 :0:0 Semaglutide Semaglutide 7.9 7.9

Com.Ex. Com. Ex.22 - - PLGA50B PLGA50B - - 00 :: 11: 00 0:1:0 Semaglutide Semaglutide 9.3 9.3

Com.Ex. Com. Ex.33 - - PLGA50B PLGA50B PLGA65B PLGA65B 0:1:1 0:1:1 Semaglutide Semaglutide 9.0 9.0

Com.Ex. Com. Ex.44 - - -- PLGA65B PLGA65B 00 : 00 :11 0:0:1 Semaglutide Semaglutide 9.1 9.1

Com. Ex.55 Com. Ex. - - PLGA50B PLGA50B PLGA65B PLGA65B 00 :: 11: 22 0:1:2 Tirzepatide Tirzepatide 11.5 11.5

[00101]

16

[00102] The ratios

[00102] The ratios of of LA and GA LA and GAininthe thehigh-viscosity high-viscosity PLGA andininthe PLGA and the total total PLGA PLGA ofof

microspheresinin Examples microspheres Examples1 1toto1010and and Comparative Comparative Examples Examples 1 to 1 5 to 5 were were as shown as shown in Table in Table 3 3

below. below.

[00103] [Table3]3]

[00103] [Table

Classification Classification LA LA:::GA LA GA GA ofhigh-viscosity of of high-viscosityPLGA high-viscosity PLGA PLGA LA:: GA LA LA: GAof GA oftotal of total PLGA total PLGA PLGA A:B:C A:B:C A:B:C Example 11 Example 50 :50 50 50 50 50 :5050 11:1:1 :1:1 Example 22 Example 57.5 57.5 : 42.5 57.5 : 42.5 42.5 55 55 :4545 1 1 11:1:1 :1:1 1

Example 33 Example 57.5 57.5:: 42.5 57.5 42.5 42.5 55 : 55 45 45 11:1:1 :1:1 Example 44 Example 57.5 57.5 :: 42.5 42.5 57.5 42.5 55 : 55 45 45 11:1:1 : 1 : 11 Example 55 Example 62.5 62.5:: 37.5 62.5 37.5 37.5 58 :42 58 42 11:1:1 :1:1 Example 66 Example 60 : 60 40 40 57.5 57.5 :: 42.5 42.5 57.5 42.5 1 1 11:1:2 : 1 : 22 Example 77 Example 63 63 :3737 60 :40 60 40 11:1:2 :1:2 1: 2 Example 88 Example 57.5 57.5 :: 42.5 42.5 57.5 42.5 56 : 56 44 44 11:2:2 : 2 : 22 1:2: Example 99 Example 53 :47 53 47 52.5 52.5 :: 47.5 47.5 11:4:1 : 44 :11 Example Example 1010 57.5 57.5 :: 42.5 42.5 57.5 42.5 55 : 55 45 45 11:1:1 :1:1 Com. Example Com. Example11 -- 50 :50 50 50 1 1 11:0:0 :0:0 Com. Example Com. Example22 50 :50 50 50 50 50 :: 50 50 50 50 0 :1:0 0:1:0 Com. Example Com. Example33 57.5 57.5:: 42.5 57.5 42.5 42.5 57.5 57.5:: 42.5 57.5 42.5 42.5 0 0 00 : :1 :11 0:1:1 Com. Example Com. Example44 65 : 65 35 35 65 : 65 35 35 00 : 00 :11 0:0:1 Com. Example 55 Com. Example 60 :40 60 40 60 :40 60 40 0:1:2 0:1:2

[00104]

[00105] 2. Measurement

[00105] 2. Measurement of diameter of diameter of sustained-release of sustained-release microsphere microsphere

[00106] Mean

[00106] Mean diameters diameters (M.D) (M.D) of Examples of Examples 1 to 1 to 10 and10Comparative and Comparative ExamplesExamples 1 to 5 were 1 to 5 were

quantitatively measured. quantitatively 180mgmg measured. 180 of of microspheres microspheres andand 10 10 mL mL of PVA of 1% 1% PVA were were added added to mL to a 50 a 50 mL

conical tube, conical tube, dispersed dispersed using an ultrasonic using an ultrasonic generator for 11 minute, generator for and then minute, and then introduced introducedinto intoaa

diameter analyzer diameter analyzer (CILAS, (CILAS,French), French),and andthen thendiameters diameterswere weremeasured measured and and shown shown in Table in Table 4. 4.

[00107] [Table4]4]

[00107] [Table

Classification Classification Diameter (m) Diameter(um) (µm) Example Example 11 Example 1 21.3 21.3 Example 22 Example 18.7 18.7

Example 33 Example 19.3 19.3

Example 44 Example 21.8 21.8 Example 55 Example 18.7 18.7

Example 66 Example 22 22 Example 77 Example 21.7 21.7

17

Example 88 Example 22.6 22.6 Example 99 Example 20.6 20.6 Example 10 Example 10 20 20 ComparativeExample Comparative Example1 1 18.5 18.5

Comparative Example Comparative Example 2 2 20.9 20.9 ComparativeExample Comparative Example3 3 21.2 21.2 ComparativeExample Comparative Example4 4 21.3 21.3 ComparativeExample Comparative Example5 5 20.1 20.1 20.1

[00108]

[00109] 3. Pharmacokinetic

[00109] 3. Pharmacokinetic profile profile of of sustained-release sustained-release formulation formulation in S.D. in S.D. rats rats

[00110]

[00110] TheThe sustained-release sustained-release formulation formulation containing containing semaglutide semaglutide or tirzepatide or tirzepatide according according to the to the

present invention was administered to 8-week-old S.D rats (n = 5), and the drug release patterns present invention was administered to 8-week-old S.D rats (n = 5), and the drug release patterns

in the body were evaluated. in the body were evaluated.

[00111] Thesustained-release

[00111] The sustained-releaseformulations formulations containing containing the the microspheres microspheres of Examples of Examples and and

ComparativeExamples Comparative Examples were were administered administered to 8-week-old to 8-week-old male rats male S.D. S.D.through rats through subcutaneous subcutaneous

injection (S.C) injection at aa dose (S.C) at doseofof2mpk 2mpk (mg/kg), (mg/kg), and then and then the blood the blood concentration concentration of the of drugthe drug

(semaglutide or (semaglutide or tirzepatide) tirzepatide) was was observed overtime. observed over time. 0.50.5 ml ml of of blood blood waswas collected collected fromfrom the the

jugular vein at 1 hour, 3 hours, 6 hours, 24 hours, 3 days, 5 days, 7 days, 9 days, 11 days, 14 days, jugular vein at 1 hour, 3 hours, 6 hours, 24 hours, 3 days, 5 days, 7 days, 9 days, 11 days, 14 days,

16 days,1818days, 16 days, days,2121 days, days, 24 24 days, days, 28 days, 28 days, 31 days, 31 days, and 35and days35 days 17 (total (total 17 points) points) after injection, after injection,

respectively, stored at -80C, and then subjected to LC-MS/MS analysis. respectively, stored at -80°C, and then subjected to LC-MS/MS analysis.

[00112] Table

[00112] Table 5 shows 5 shows relative relative AUC and AUC values values and cumulative cumulative release release rates rates of (the ratio (the ratio of cumulative cumulative

drug release over time to the total amount released). The bioavailability was evaluated based on a drug release over time to the total amount released). The bioavailability was evaluated based on a

relative AUC value, calculated as the ratio of the AUC of the test formulation to the AUC of drug- relative AUC value, calculated as the ratio of the AUC of the test formulation to the AUC of drug-

only administration, only administration, after aftercalculating the the calculating areaarea under thethecurve under curve(AUC) (AUC) from the blood from the blood

concentration-time profile. The initial burst release and delayed release (lag phase) phenomena concentration-time profile. The initial burst release and delayed release (lag phase) phenomena

were evaluated using the cumulative release rate according to the following evaluation criteria. were evaluated using the cumulative release rate according to the following evaluation criteria.

[00113] * Evaluation

[00113] * Evaluation criteria criteria

[00114]

[00114] 1) 1) Initialburst Initial burst release release evaluation: evaluation: In case In the the case of 5% of or 5% less or less 1within within day (241 hours), day (24it hours), it

18 was evaluated that there was no initial burst release. was evaluated that there was no initial burst release.

[00115] 2)Lag

[00115] 2) Lagphase phaseevaluation: evaluation:In In the the case case of of 10% or more 10% or by 11 week more by week(day (day7), 7), 20% 20%orormore morebyby

day 99 (or day (or 25% ormore 25% or morebybyday day10), 10),and and50% 50% or or more more by 2byweeks 2 weeks (day (day 14), 14), it was it was evaluated evaluated thatthat

there was there was no no delayed release problem. delayed release problem.

[00116] 3)Bioavailability

[00116] 3) Bioavailability evaluation: evaluation: IfIf the the relative relative AUC AUC value value waswas 50% 50% or more, or more, it was it was

evaluated that the bioavailability was high. evaluated that the bioavailability was high.

[00117]

[00118] Thesustained-release

[00118] The sustained-releaseformulations formulations of of Examples Examples 1 to 1 10 to 10 according according to the present to the present

invention exhibited invention exhibited relative relativeAUC AUC values values of of 50% or more 50% or (Table 55 and more (Table and FIGS. 1A,1B, FIGS. 1A, 1B,and and1C), 1C), and and

were evaluated to have high bioavailability according to the evaluation criteria. In addition, in were evaluated to have high bioavailability according to the evaluation criteria. In addition, in

the case of initial release, the initial cumulative release was 5% or less, so that there was no initial the case of initial release, the initial cumulative release was 5% or less, SO so that there was no initial

burst release, and the cumulative release rate by day 7 was 10% or more, the cumulative release burst release, and the cumulative release rate by day 7 was 10% or more, the cumulative release

rate by rate by day day 99 was was20% 20%or or more, more, andand thethe cumulative cumulative release release rate rate by by dayday 14 was 14 was 50% 50% or or more, more,

confirming that the sustained-release formulations had no delayed release (lag phase) issue (Table confirming that the sustained-release formulations had no delayed release (lag phase) issue (Table

5 and FIG. 2). 5 and FIG. 2).

[00119] However,

[00119] However, in in thecase the caseofofthe thesustained-release sustained-release microsphere microsphereformulation formulationofofComparative Comparative

Example1,1,the Example thebioavailability bioavailability was high and was high andthere therewas wasnonodelayed delayed releaseissue, release issue,but butunlike unlikethe the

Examples,there Examples, there was wasa aproblem problem thatthe that theinitial initial release release was excessively high, was excessively high, and and in in the the case case of of

Comparative Examples 2 to 5, there was no initial burst release, but there was a problem of low Comparative Examples 2 to 5, there was no initial burst release, but there was a problem of low

bioavailability and delayed release lasting more than one week after administration (Table 5 and bioavailability and delayed release lasting more than one week after administration (Table 5 and

FIG. 1D). FIG. 1D).

[00120] [Table5]5]

[00120] [Table

Cumulative release rate (ratio of cumulative release over time to Cumulative release rate (ratio of cumulative release over time to Relative Relative Classification Classification total release total amount) release amount) AUC AUC Day11 Day Day 77 Day Day 99 Day Day 14 Day 14 Day 21 Day 21 Day Day 28 28 (%) (%)

19

Ex. 1 Ex. 1 1.8% 1.8% 22.9% 22.9% 42.9% 42.9% 74.2% 74.2% 74.2% 97.3% 97.3% 99.9% 99.9% 99.9% 53.5% 53.5% Ex. 22 Ex. 1.2% 1.2% 20.3% 20.3% 20.3% 34.5% 34.5% 34.5% 64.1% 64.1% 91.0% 91.0% 99.3% 99.3% 99.3% 52.0% 52.0% Ex. 33 Ex. 0.8% 0.8% 0.8% 25.6% 25.6% 37.1% 37.1% 69.0% 69.0% 95.2% 95.2% 99.8% 99.8% 99.8% 60.9% 60.9% Ex. 44 Ex. 4.6% 4.6% 32.8% 32.8% 32.8% 47.7% 47.7% 74.3% 74.3% 96.3% 96.3% 99.8% 99.8% 68.4% 68.4% Ex. 55 Ex. 2.5% 2.5% 26.5% 26.5% 26.5% 37.3% 37.3% 59.7% 59.7% 84.6% 84.6% 97.0% 97.0% 63.1% 63.1% Ex. 66 Ex. 1.1% 1.1% 13.8% 13.8% 26.0% 26.0% 26.0% 53.4% 53.4% 87.5% 87.5% 99.4% 99.4% 51.1% 51.1% Ex. 77 Ex. 1.6% 1.6% 17.8% 17.8% 32.9% 32.9% 59.9% 59.9% 83.0% 83.0% 97.2% 97.2% 97.2% 53.0% 53.0% Ex. 88 Ex. 1.7% 1.7% 16.8% 16.8% 31.4% 31.4% 62.8% 62.8% 91.3% 91.3% 91.3% 99.5% 99.5% 55.4% 55.4% Ex. 99 Ex. 1.6% 1.6% 15.2% 15.2% 27.7% 27.7% 27.7% 61.8% 61.8% 95.4% 95.4% 99.9% 99.9% 55.7% 55.7% Ex. 10 Ex. 10 1.5% 1.5% 36.8% 36.8% 36.8% 43.9% 43.9% 67.1% 67.1% 92.1% 92.1% 99.7% 99.7% 67.4% 67.4% Com.Ex. Com. Ex.11 6.8% 6.8% 62.4% 62.4% 62.4% 79.6% 79.6% 98.7% 98.7% 98.7% 100.0% 100.0% 100.0% 100.0% 56.8% 56.8% Com.Ex. Com. Ex.22 1.3% 1.3% 7.8% 7.8% 7.8% 16.8% 16.8% 56.6% 56.6% 94.4% 94.4% 94.4% 99.9% 99.9% 99.9% 32.0% 32.0% Com.Ex. Com. Ex.33 0.7% 0.7% 3.7% 3.7% 6.6% 6.6% 6.6% 21.5% 21.5% 55.6% 55.6% 82.6% 82.6% 46.4% 46.4% 46.4% Com.Ex. Com. Ex.44 1.8% 1.8% 7.7% 7.7% 10.5% 10.5% 39.3% 39.3% 39.3% 67.5% 67.5% 94.4% 94.4% 28.5% 28.5% Com.Ex. Com. Ex.55 0.8% 0.8% 5.1% 5.1% 12.6% 12.6% 25.9% 25.9% 25.9% 61.5% 61.5% 86.9% 86.9% 45.8% 45.8% 45.8%

[00121]

[00122] Inaddition,

[00122] In addition, since since Examples Examples1 1to to1010 showed showed similar similar release release patterns, patterns, Example Example 3 was 3 was

selected as a representative example as shown in FIG. 2. It may be seen that Examples according selected as a representative example as shown in FIG. 2. It may be seen that Examples according

to the to the present present invention invention showed sustainedrelease showed sustained releasewithout withouta adelayed delayedrelease releaseissue issueand andwithout without

excessive initial excessive initial release, while release, Comparative while ComparativeExample 1, in Example 1, in which only low-viscosity which only low-viscosity PLGA PLGA waswas

used, showed used, showedinitial initialburst burstrelease releaseand anda short a short release release period period compared compared to Examples, to Examples, and and

ComparativeExamples Comparative Examples 2 to 2 to 4, 4, ininwhich which only only high-viscosityPLGA high-viscosity PLGA was was used,used, showed showed a delayed a delayed

release phenomenon release lastingmore phenomenon lasting morethan thanone oneweek. week.

[00123] Subsequent

[00123] Subsequent experiments experiments were were also also conducted conducted by by selecting selecting Example Example 3 as3 as a representative a representative

example. example.

[00124]

[00125] 4. Pharmacokinetic

[00125] 4. Pharmacokinetic profile profile of of sustained-release sustained-release formulation formulation in minipigs in minipigs

[00126] Next,

[00126] Next, in order in order to evaluate to evaluate the pharmacokinetics the pharmacokinetics in medium-sized in medium-sized animals, animals, the the sustained- sustained-

release formulation of Example 3 according to the present invention was administered to minipigs, release formulation of Example 3 according to the present invention was administered to minipigs,

and the drug release patterns within the microspheres in the body were evaluated. and the drug release patterns within the microspheres in the body were evaluated.

[00127] Aftersubcutaneous

[00127] After subcutaneousinjection injection(S.C) (S.C)of of drug drug at at aa dose dose of of 0.08 0.08 mpk (mg/kg)into mpk (mg/kg) into20 20toto 30 30

20 kg of kg of male SPFminipigs, male SPF minipigs,the theblood bloodconcentration concentrationofofsemaglutide semaglutidewas wasobserved observed over over time.A A time. 2.5 2.5 ml of blood was collected from the jugular vein at 1 hour, 6 hours, 12 hours, 24 hours, 2 days, 3 ml of blood was collected from the jugular vein at 1 hour, 6 hours, 12 hours, 24 hours, 2 days, 3 days, 5 days, 7 days, 9 days, 11 days, 14 days, 16 days, 18 days, 21 days, 24 days, 28 days, 31 days, 5 days, 7 days, 9 days, 11 days, 14 days, 16 days, 18 days, 21 days, 24 days, 28 days, 31 days, 35 days, 42 days, 49 days, 56 days, 63 days, and 70 days (total 23 points) after injection, days, 35 days, 42 days, 49 days, 56 days, 63 days, and 70 days (total 23 points) after injection, respectively, stored at -80C, and then subjected to LC-MS/MS analysis. respectively, stored at -80°C, and then subjected to LC-MS/MS analysis.

[00128] Theresults

[00128] The results may maybebeseen seenininFIGS. FIGS.3 3and and4,4,where whereFIG. FIG. 3 shows 3 shows thethe blood blood concentration concentration

of semaglutide and FIG. 4 shows the cumulative release rate of semaglutide. Through the results of semaglutide and FIG. 4 shows the cumulative release rate of semaglutide. Through the results

according to according to the the present present invention, invention, it it was confirmedthat was confirmed that there there was wasnonoinitial initial burst burst release release or or

delayed release in minipigs, an animal model more similar to humans than rats, and that the release delayed release in minipigs, an animal model more similar to humans than rats, and that the release

duration of semaglutide was longer in minipigs, which are larger than S.D. rats. In addition, it duration of semaglutide was longer in minipigs, which are larger than S.D. rats. In addition, it

may be inferred that the release duration will further increase when administered to humans. may be inferred that the release duration will further increase when administered to humans.

[00129]

[00130] 5. Evaluation

[00130] 5. Evaluationofofanti-obesity anti-obesityefficacy efficacy

[00131] Thesustained-release

[00131] The sustained-releaseformulation formulationofofExample Example 3 according 3 according to to thepresent the presentinvention inventionwas was

administered to a diet-induced obesity (DIO) mouse model to confirm the anti-obesity efficacy. administered to a diet-induced obesity (DIO) mouse model to confirm the anti-obesity efficacy.

[00132] Sixteen-week-old male

[00132] Sixteen-week-old male C57BL/6J C57BL/6J and and C57BL/6J-DIO mice(The C57BL/6J-DIO mice (TheJackson Jackson Laboratory Laboratory

(JAX), Saeron Bio) were used in the study. After introduction, the mice were sufficiently fed with (JAX), Saeron Bio) were used in the study. After introduction, the mice were sufficiently fed with

a regular chow diet (5 kcal % of fat, PicoLab, #5053, USA) or a high fat diet (60 kcal % of fat, a regular chow diet (5 kcal % of fat, PicoLab, #5053, USA) or a high fat diet (60 kcal % of fat,

Researchdiet, Research diet, D12492, USA)together D12492, USA) togetherwith withwater, water,maintained maintainedatataa temperature temperatureof of 22 22 H±± 2°C 2Cand andaa

humidity of humidity of 50 50 +±± 10%, 10%,and andacclimated acclimatedininindividual individualcages cagesunder underananautomatically automaticallycontrolled controlled12- 12-

hour light/dark cycle (dark: 7 pm to 7 am). After one week of acclimation, all prepared subjects hour light/dark cycle (dark: 7 pm to 7 am). After one week of acclimation, all prepared subjects

were administered were administeredand andacclimated, acclimated, andand clinical clinical signs signs (activity,stool (activity, stool abnormalities, abnormalities, bleeding, bleeding,

wounds,deformities, wounds, deformities,skin, skin,hair, hair, eye eyeabnormalities, abnormalities,etc.) etc.) were wereobserved observed twice twice a (when a day day (when

administered in administered in the the morning and when morning and whenmonitored monitoredininthe theafternoon). afternoon).

21

[00133] Toevaluate

[00133] To evaluatebody bodyweight weight and and food food intake,the intake, thedrug drugwas wasadministered administered for8 8weeks for weeks under under

conditions described conditions described in in Table Table 6, 6,and andbody body weight weight (FX-2000i, (FX-2000i, A&D company, A&D company, 0.01 0.01 to to 2200 2200 g) g) andand

food intake food intake (CSG201F, OHAUS, (CSG201F, OHAUS, 0.1 g0.1 tog200 to 200 g) were g) were measured measured daily daily starting starting fromfrom the first the first dayday

of administration. of administration. AllAllresult resultvalues valueswere were expressed expressed as the as the meanmean I ± standard ± standard error,error, and and the the

statistical significance of the group was calculated through an independent t-test between test statistical significance of the group was calculated through an independent t-test between test

substance administration substance administration groups. The groups. The resultswere results wereshown shownin in Table6.6. Table

[00134] [Table6]6]

[00134] [Table

Group(N(N==8)8) Group Diet Diet Administration Administration Drug Drug Drug Administration Administration route route route concentration concentration cycle cycle (mpk) (mpk) 1. 1. Normal (vehicle) Normal (vehicle) RC(Regular RC (Regular S.C. S.C. - - QD QD chow) chow) 2. Negative control (DIO, 2. 2. Negative Negativecontrol (DIO, control (DIO, 60% HFD 60% HFD S.C. S.C. - QD - QD Con(-),vehicle) Con(-), vehicle) (High fat (High fat

diet) diet) diet)

3. Positive control (DIO, 3. Positive control (DIO, 60% HFD 60% HFD S.C. S.C. 0.1 0.1 QD QD Con(+), semaglutide) Con(+), semaglutide) QD 4. Example 4. Example 33 60% HFD 60% HFD S.C. S.C. 4 4 Q3D Q3D 5. 5. Example Example 33 60% HFD 60% HFD S.C. S.C. 4 4 QW 6. Example 6. Example 33 60% HFD S.C. S.C. 4 4 QW Q2W 60% HFD Q2W Q2W

[00135]

[00136] Thebody

[00136] The body weight weight measurement measurement results results were were shown shown in FIG. in FIG. 5A, 5A, and and the the body body weight weight gaingain

rate (Body rate (ABody weightfrom weight (Body weight from from baseline(vs baseline baseline (vsCon(-)), (vs Con(-)), %) Con(-)), %) was %) was shown was shown shown inFIG. in in FIG.5B. FIG. 5B.This 5B. This This waswas was shown shown shown as the as the as the

bodyweight body weightchange change(%)(%) calculatedbyby calculated comparing comparing the the drug-administered drug-administered groups groups to the to the negative negative

control group. control group.

[00137] From

[00137] From FIG. FIG. 5, 5, ititwas wasfound foundthat thatthe the normal normalcontrol control group groupand andthe thenegative negativecontrol control group group

increased in increased in body weight, whereas body weight, whereasthe the positive positive control control group and Example group and Example3 3(Q3D, (Q3D, QW,QW, Q2W)Q2W)

showeda astatistically showed statistically significant significantdecrease decrease(p(p< <0.05) 0.05)ininbody body weight weight compared tothe compared to the negative negative

control group. In addition, referring to FIG. 5, it was found that the positive control group and control group. In addition, referring to FIG. 5, it was found that the positive control group and

Example3 3(Q3D, Example (Q3D,QW,QW, Q2W)Q2W) showedshowed a statistically a statistically significant significant (p0.05) (p < < 0.05) body body weight weight lossloss raterate

22 compared compared compared to totothethe the negative negative control control negative group. group. control group.

[00138]

[00138] In In addition, addition, thethe food food intake intake (g) shown (g) was was shown in FIG. in 6. FIG. 6.positive In the In thecontrol positive control group and group and

all evaluated all evaluated drug drug administration administration groups, groups, the the food intake measured food intake for the measured for the drug drugadministration administration

period rapidly decreased immediately after drug administration and then gradually recovered, but period rapidly decreased immediately after drug administration and then gradually recovered, but

the decreased the decreasedfood foodintake intakewaswas maintained maintained compared compared to the to the negative negative control control group, group, and a and a

statistically significant decrease (p < 0.05) in food intake was observed compared to the negative statistically significant decrease (p < 0.05) in food intake was observed compared to the negative

control group. control group.

[00139] From

[00139] From these these results, results, it was it was found found that,that, compared compared to the to the positive positive controlcontrol group administered group administered

daily, the sustained-release formulation of the present invention was administered to exhibit a daily, the sustained-release formulation of the present invention was administered to exhibit a

significant weight loss effect with only one administration every two weeks (based on mice). significant weight loss effect with only one administration every two weeks (based on mice).

[00140] Insummary,

[00140] In summary,it it waswas confirmed confirmed that that the sustained-release the sustained-release formulation formulation of present of the the present

invention was a formulation that had no issue of initial burst release or delayed release of the drug, invention was a formulation that had no issue of initial burst release or delayed release of the drug,

enabled long-term sustained release, and had high bioavailability. enabled long-term sustained release, and had high bioavailability.

23

Claims

[CLAIMS]

[CLAIMS]

[Claim 1]

[Claim 1]

A sustained-release A sustained-release microsphere comprising: microsphere comprising:

a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof and polylactide- a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof and polylactide-

co-glycolide (PLGA), co-glycolide (PLGA),

wherein the wherein the GLP-1 GLP-1 receptor receptor agonist agonist or or thethe pharmaceutically pharmaceutically acceptable acceptable salt salt thereof thereof is is

included in an amount of 3 to 12 wt% of the total wt%, and included in an amount of 3 to 12 wt% of the total wt%, and

wherein the wherein the PLGA PLGA is mixture is a a mixture of one of one typetype of low-viscosity of low-viscosity PLGA PLGA with anwith an intrinsic intrinsic

viscosity of viscosity of 0.14 0.14 to to 0.24 0.24 dL/g dL/g and two or and two or more moretypes typesofofhigh-viscosity high-viscosity PLGA PLGA with with an an intrinsic intrinsic

viscosity of 0.32 to 0.60 dL/g. viscosity of 0.32 to 0.60 dL/g.
[Claim 2]

[Claim 2]

The sustained-release The sustained-release microsphere microsphereofofclaim claim1, 1,wherein wherein thethe oneone typetype of low-viscosity of low-viscosity

PLGA PLGA and and thethe two two or or more more types types of of high-viscosity high-viscosity PLGA PLGA in the in the mixture mixture have have a weight a weight ratioratio of of

1:0.2 1:0.2 toto5.5.
[Claim 3]

[Claim 3]

The sustained-release The sustained-release microsphere microsphereofofclaim claim1,1,wherein wherein thethe GLP-1 GLP-1 receptor receptor agonist agonist is is

semaglutide or tirzepatide. semaglutide or tirzepatide.
[Claim 4]

[Claim 4]

The sustained-release The sustained-release microsphere microsphereofofclaim claim1,1,wherein whereinthe thetwo twoor ormore more types types of of high- high-

viscosity PLGA are one or more types of PLGA with a molar ratio of lactide to glycolide of 45 to viscosity PLGA are one or more types of PLGA with a molar ratio of lactide to glycolide of 45 to

55 :: 45 to 55 45 to andone 55 and oneorormore more types types of PLGA of PLGA with a with molara ratio molarofratio of lactide lactide to glycolide to glycolide of 60 to of 8060 : to 80 :

20 to 40. 20 to 40.
[Claim 5]

[Claim 5]

The sustained-release microsphere of claim 1, wherein the poly(lactide-co-glycolide) is The sustained-release microsphere of claim 1, wherein the poly(lactide-co-glycolide) is

24 included in an amount of 88 to 97 wt% of the total weight of the sustained-release microsphere. included in an amount of 88 to 97 wt% of the total weight of the sustained-release microsphere.
[Claim 6]

[Claim 6]

The sustained-release microsphere of claim 1, wherein the sustained-release microsphere The sustained-release microsphere of claim 1, wherein the sustained-release microsphere

is prepared is by dissolving prepared by dissolving the the GLP-1 GLP-1 receptor receptor agonist agonist or or thethe pharmaceutically pharmaceutically acceptable acceptable saltsalt

thereof and the poly(lactide-co-glycolide) in glacial acetic acid to prepare a mixed solution; and thereof and the poly(lactide-co-glycolide) in glacial acetic acid to prepare a mixed solution; and

spraying the mixed solution using ultrasonic spray nozzles, and then evaporating the glacial acetic spraying the mixed solution using ultrasonic spray nozzles, and then evaporating the glacial acetic

acid using dry air. acid using dry air.
[Claim 7]

[Claim 7]

The sustained-release The sustained-release microsphere microsphereofofclaim claim6,6,wherein whereinthethefrequency frequency of of thethe ultrasonic ultrasonic

spray nozzles is 40 to 80 kHz. spray nozzles is 40 to 80 kHz.
[Claim 8]

[Claim 8]

The sustained-release microsphere of claim 1, wherein the sustained-release microsphere The sustained-release microsphere of claim 1, wherein the sustained-release microsphere

is administered to S.D rats or minipigs and then the GLP-1 receptor agonist or the pharmaceutically is administered to S.D rats or minipigs and then the GLP-1 receptor agonist or the pharmaceutically

acceptable salt thereof is released at less than 5% within 24 hours, and released at 10% or more acceptable salt thereof is released at less than 5% within 24 hours, and released at 10% or more

within 11 week, within 20%orormore week, 20% morewithin within9 9days, days,or or 50% 50%orormore morewithin within2 2weeks. weeks.
[Claim 9]

[Claim 9]

The sustained-release microsphere of claim 1, wherein the sustained-release microsphere The sustained-release microsphere of claim 1, wherein the sustained-release microsphere

is administered at intervals of one to three months. is administered at intervals of one to three months.
[Claim 10]

[Claim 10]

The sustained-release microsphere of claim 1, wherein the sustained-release microsphere The sustained-release microsphere of claim 1, wherein the sustained-release microsphere

has aa mean has diameterof mean diameter of 15 15 to 25 m. to 25 um. µm.
[Claim 11]

[Claim 11]

A sustained-release formulation comprising the sustained-release microsphere of any one A sustained-release formulation comprising the sustained-release microsphere of any one

of claims 1 to 10. of claims 1 to 10.

25
[Claim 12]

[Claim 12]

A pharmaceutical composition for treating or preventing diabetes, obesity, non-alcoholic A pharmaceutical composition for treating or preventing diabetes, obesity, non-alcoholic

steatohepatitis or degenerative brain disease, comprising the sustained-release microsphere of any steatohepatitis or degenerative brain disease, comprising the sustained-release microsphere of any

one of claims 1 to 10. one of claims 1 to 10.
[Claim 13]

[Claim 13]

The pharmaceutical The pharmaceuticalcomposition compositionof of claim claim 12,12, wherein wherein thethe degenerative degenerative brain brain disease disease is is

any one any oneselected selectedfrom from the the group group consisting consisting of Parkinson’s of Parkinson's disease, disease, Alzheimer’s Alzheimer's disease, disease,

Huntington’s disease, Amyotrophic lateral sclerosis(ALS), Creutzfeldt-Jakob disease, stroke, and Huntington's disease, Amyotrophic lateral sclerosis(ALS) sclerosis(ALS),Creutzfeldt-Jakob Creutzfeldt-Jakobdisease, disease,stroke, stroke,and and

multiple sclerosis. multiple sclerosis.
[Claim 14]

[Claim 14]

A method for producing a sustained-release microsphere or sustained-release formulation A method for producing a sustained-release microsphere or sustained-release formulation

comprising: comprising:

dissolving 3 to 12 wt% of a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof dissolving 3 to 12 wt% of a GLP-1 receptor agonist or a pharmaceutically acceptable salt thereof

and 88 and 88 toto 97 97wt% wt%of of poly(lactide-co-glycolide)inina asolvent poly(lactide-co-glycolide) solventtotoprepare preparea amixed mixed solution; solution; andand

spraying the mixed solution using ultrasonic spray nozzles, and then evaporating the solvent using spraying the mixed solution using ultrasonic spray nozzles, and then evaporating the solvent using

dry air to prepare microspheres, dry air to prepare microspheres,

wherein the wherein the poly(lactide-co-glycolide) poly(lactide-co-glycolide) is is aa mixture mixture of of one one type type of of low-viscosity low-viscosity PLGA PLGA

with an intrinsic viscosity of 0.14 to 0.24 dL/g and two or more types of high-viscosity PLGA with with an intrinsic viscosity of 0.14 to 0.24 dL/g and two or more types of high-viscosity PLGA with

an intrinsic viscosity of 0.32 to 0.60 dL/g. an intrinsic viscosity of 0.32 to 0.60 dL/g.

26

[FIG. 1A]

[FIG. 1A]

600

500 500 (ng/ml) concentration (ng/ml) concentration 400 400

300 300

200 200

100 * X

0 0 7 14 21 28 35 Day Example2 Example3 Example3 Example4 Example10

1/6 1/6

[FIG. 1B]

[FIG. 1B]

500

400 400 (ng/ml) concentration (ng/ml) concentration 300

200 200

100

0 0 0 7 14 21 28 35 35 Day Example 1 Example 5 Example 7

[FIG. 1C]

[FIG. 1C]

400

300 (ng/ml) concentration (ng/ml) concentration 200 200

100

0 0 7 14 21 28 35 Day Example6 Example 8 Example 9

2/6 2/6

[FIG. 1D]

[FIG. 1D]

500 500

400 concentration concentration (ng/ml) (ng/ml) 300 300

200

100

* *

0 0 7 14 21 28 35

Comparative Comparative Day Comparative Comparative Example 1 Example 2 Example 3 Example 4

3/6 3/6

[FIG. 2]

[FIG. 2]

100% (%) rate release Cumulative (%) rate release Cumulative 80%

60%

40%

20%

0% 0 7 14 14 21 28 35

Comparative DAY Comparative Comparative Comparative Example3 Example 1 Example 3 Example 4 Example 2

[FIG. 3]

[FIG. 3]

90

80

70 Example3 Example3 (ng/ml) concentration (ng/ml) concentration 60

50

40

30

20

10

0 0 7 7 14 21 28 35 42 49 56 63 70 Day Day

4/6 4/6

[FIG. 4]

[FIG. 4]

100% (%) rate release Cumulative (%) rate release Cumulative 80% Example3 Example3 60%

40%

20%

0% 0 7 14 21 28 35 42 49 56 63 70 Day

[FIG. 5A]

[FIG. 5A]

60

Body weight (g)

Nor, Normal, QD 50 DIO, Negative Control, QD

DIO, Positive Control, QD

40 DIO, Example 3, Q3D

DIO, Example 3, QW

DIO, Example 3, Q2W 30

20 0 7 14 21 28 35 42 49 56 Time (Day)

5/6 5/6 from change weight Body from change weight Body

[FIG. 5B]

[FIG. 5B]

10 Con(-)) vs (%, baseline Con(-)) vs (%, baseline 0

-10 Nor, Normal, QD DIO, Negative Control, QD

-20 -20 DIO, Positive Control, QD

DIO, Example 3, Q3D

-30 DIO, Example 3, QW

DIO, Example 3, Q2W

-40

-50 0 7 14 21 28 35 42 49 56 Time (Day)

[FIG. 6]

[FIG. 6] (kcal) intake food Cumulative (kcal) intake food Cumulative 1000

800 Nor, Normal, QD DIO, Negative Control, QD 600 DIO, Positive Control, QD

DIO, Example 3, Q3D 400 DIO, Example 3, QW

DIO, Example 3, Q2W 200

0 0 7 14 21 28 35 42 49 56 Time (Day)

6/6 6/6