KR20240173155A - Pharmaceutical composition comprising a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptors triagonist - Google Patents

Abstract

The present invention provides a pharmaceutical composition comprising: microspheres including at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist; and at least one selected from a first carrier and a water-soluble liquid, wherein the first carrier includes a fluid oil including a fatty acid ester, a surfactant, or a polyhydric alcohol. The pharmaceutical composition maintains stability of the microspheres in the suspension formulation for a long period of time, and exhibits excellent elution characteristics of the microspheres stored for a long period of time. In addition, the pharmaceutical composition exhibits an effect of increasing bioavailability and reducing the administration dose.

Description

{Pharmaceutical composition comprising a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptors triagonist}

The present invention relates to a pharmaceutical composition comprising a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptor triple agonist.

Glucagon-like peptide-1 (GLP-1) is derived from pre-proglucagon, a 158 amino acid precursor polypeptide that is processed in different tissues to form numerous different proglucagon-derived peptides, including glucagon, glucagon-like peptide-1 (GLP-1), glucagon-like peptide-2 (GLP-2) and oxyntomodulin (OXM), which are involved in regulation of food intake as well as various physiological functions, including glucose homeostasis, insulin secretion, gastric emptying and intestinal growth.

GLP-1 is produced as a 37-amino acid peptide corresponding to amino acids 72 to 108 of proglucagon (preproglucagon 92 to 128). GLP-1(7-36) amide or GLP-1(7-37) acid are the biologically active forms of GLP-1 which exhibit essentially identical activity at the GLP-1 receptor. GLP-1 and GLP-1 analogues that act as agonists at the GLP-1 receptor have been shown to provide effective glycemic control, for example, for the treatment of patients with type 2 diabetes, and in addition provide weight loss, preservation of beta-cell function, and alleviation of hypertension, hypoglycemia and/or hyperlipidemia.

Currently, certain GLP-1 analogues are marketed or in development, including Byetta ® & Bydureon BCise ® (exenatide), Ozempic ® (semaglutide), Victoza ® (liraglutide), Adlyxin ® (lixisenatide), Tanzeum ® (albiglutide), and Trulicity ® (dulaglutide).

Like GLP-1, GIP (Gastric Inhibitory Polypeptide) not only stimulates insulin secretion in a glucose-dependent manner in beta cells, but also promotes insulin synthesis, induces beta cell proliferation, and has an inhibitory effect on apoptosis [Trumper et al., Mol Endocrinol 15:1559-1570, 2001].

Currently, GLP-1/GIP dual agonists are also being developed, for example, Mounjaro ® (tirzepatide) is marketed.

GCG receptors regulate the action of glucagon in the liver. Glucagon promotes gluconeogenesis in the liver to increase blood sugar levels, and GCG receptors inhibit the action of glucagon to maintain blood sugar levels at a constant level. Currently, GLP-1/GIP/GCG receptor triple agonists are being developed.

Administration of GLP-1 receptor agonists, GLP-1/GIP receptor agonists, and/or GLP-1/GIP/GCG receptor triple agonists is mainly administered as an injection due to several barriers such as enzymatic degradation in the gastrointestinal tract and intestinal mucosa, insufficient absorption from the intestinal mucosa, and first pass metabolism in the liver.

Extended-release injectable formulations of GLP-1 receptor agonists, GLP-1/GIP receptor agonists and/or GLP-1/GIP/GCG receptor triple agonists can provide therapeutic doses of the active ingredient for a long period of time in a single injection, thereby eliminating the need for daily injections. Commonly used extended-release injectable formulations are prepared, for example, by including microspheres and aqueous carriers.

However, since the microspheres do not have long-term stability in an aqueous carrier, they have the disadvantage that the microspheres and the aqueous carrier must be packaged and stored separately. In addition, the patient must perform several steps to combine the microspheres and the aqueous carrier before injection administration, which is very inconvenient.

Republic of Korea Publication Patent No. 10-2020-0044016

The present invention has been devised to solve the above problems of the prior art.

The object is to provide a pharmaceutical composition in which microspheres comprising a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptor triple agonist in a suspension can maintain long-term stability.

In addition, it is an object of the present invention to provide a pharmaceutical composition in which the microparticles stored in a suspension for a long period of time can provide excellent dissolution characteristics.

In addition, the present invention aims to provide a pharmaceutical composition of microspheres containing a surfactant or polyhydric alcohol as a carrier to increase bioavailability.

To solve the above problem,

The present invention provides a pharmaceutical composition comprising microparticles comprising at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptor triple agonist; and at least one selected from a first carrier and a water-soluble liquid, wherein the first carrier comprises a fluid oil comprising a fatty acid ester, a surfactant or a polyhydric alcohol.

In addition, the present invention provides a pharmaceutical composition wherein the fluid oil is at least one selected from coconut oil, palm oil, palm kernel oil, sesame oil, soybean oil, almond oil, rapeseed oil, corn oil, sunflower oil, peanut oil, olive oil, castor oil, safflower oil, cottonseed oil, and ethyl oleate.

In addition, the present invention provides a pharmaceutical composition, wherein the surfactant is at least one selected from Poloxamer 188, Poloxamer 407, Polysorbate 20, Polysorbate 60, Polysorbate 80, Polyoxyethylene oleyl ether, Polyoxyethylene cetyl ether, Polyoxyethylene stearyl ether, and Polyoxyethylene lauryl ether.

In addition, the present invention provides a pharmaceutical composition wherein the polyhydric alcohol is at least one selected from ethylene glycol, propylene glycol, polyethylene glycol, glycerol, erythritol, threitol, xylitol, ribitol, mannitol, sorbitol, and maltitol.

In addition, the present invention provides a pharmaceutical composition wherein the polyhydric alcohol is ethylene glycol, propylene glycol, polyethylene glycol or glycerol.

In addition, the present invention provides a pharmaceutical composition comprising at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist contained in microspheres in a concentration range of 0.01 mg to 2,000 mg based on 1 mL of the first carrier.

In addition, the present invention provides a pharmaceutical composition, wherein the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and/or GLP-1/GIP/GCG receptor triple agonist is at least one selected from exenatide, liraglutide, laxenatide, albiglutide, dulaglutide, semaglutide, tirzepatide, cotadutide, taspoglutide, retatrutide, and pharmaceutically acceptable salts thereof.

In addition, the present invention provides a pharmaceutical composition comprising at least one selected from the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist, and GLP-1/GIP/GCG receptor triple agonist in an amount of 3 to 50 wt% based on the total weight of microparticles.

In addition, the present invention provides a pharmaceutical composition wherein the microparticles further comprise a biocompatible polymer.

In addition, the present invention provides a pharmaceutical composition wherein the biocompatible polymer is at least one selected from polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, copolymer of lactic acid and caprolactone, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid, and copolymer of lactic acid and amino acid.

In addition, the present invention provides a pharmaceutical composition comprising the biocompatible polymer in an amount of 50 to 97 wt% based on the total weight of the microparticles.

In addition, the present invention provides a pharmaceutical composition further comprising at least one additive selected from a buffer, an isotonic agent, a preservative, and a pH adjusting agent.

In addition, the present invention provides a pharmaceutical composition comprising the microspheres and at least one selected from the GLP-1 receptor agonist, the GIP/GLP-1 receptor agonist, and the GLP-1/GIP/GCG receptor triple agonist and the biocompatible polymer in a weight ratio of 1:1 to 50.

In addition, the present invention provides a pharmaceutical composition in which the microparticles remain stable for 6 months or longer.

In addition, the present invention provides a pharmaceutical composition which is an injection.

The pharmaceutical composition of the present invention provides the effect of maintaining long-term stability of microspheres comprising a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptor triple agonist in a suspension.

In addition, the microspheres stored in the suspension for a long period of time provide the effect of exhibiting excellent dissolution characteristics.

In addition, the surfactant or polyhydric alcohol according to the present invention can increase bioavailability and provide an effect of reducing the administration dosage by using microspheres as a carrier.

Figure 1 is a graph showing the in vivo pharmacokinetics of semaglutide manufactured in Example 6 and Comparative Example 2.

Hereinafter, the present invention will be described in more detail.

All technical terms used in the present invention, unless otherwise defined, are used with the same meaning as commonly understood by those skilled in the art in the relevant field of the present invention. In addition, those similar or equivalent to those described as preferred methods or samples in the present invention are also included in the scope of the present invention. The contents of all publications cited as references in this specification are incorporated herein by reference in their entirety.

The present invention relates to a pharmaceutical composition comprising microspheres comprising at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and a GLP-1/GIP/GCG receptor triple agonist; and at least one selected from a first carrier and a water-soluble liquid, wherein the first carrier comprises a fluid oil comprising a fatty acid ester, a surfactant or a polyhydric alcohol.

Improving the ease of administration of GLP-1 receptor agonists, GIP/GLP-1 receptor agonists, and/or GLP-1/GIP/GCG receptor triple agonists is a challenge that researchers in this field always face. In response to this challenge, sustained-release microspheres containing the above drugs have been developed. The sustained-release microspheres have significantly improved the convenience of administration for patients by drastically reducing the number of injections.

The above sustained-release microspheres are suspended in an aqueous carrier and injected into the human body in the form of a suspension. However, there was a problem that it was difficult to manufacture and distribute them as a suspension formulation because the stability of the sustained-release microspheres in the aqueous carrier was low. Therefore, the microspheres and the aqueous carrier are currently distributed in a separate state, and medical staff or patients mix them to manufacture and use a suspension formulation when injecting. The manufacture of the above suspension formulation is not only cumbersome, but also causes the problem that the concentration of the drug cannot be accurately adjusted during the manufacturing process of the suspension formulation when the patient directly injects it.

The pharmaceutical composition of the present invention has the characteristics that the microspheres containing a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptor triple agonist in a suspension maintain long-term stability, and the microspheres stored for a long period of time exhibit excellent dissolution characteristics.

In addition, the pharmaceutical composition of the present invention has the characteristics of increasing bioavailability and reducing the administration dosage by including a fluid oil, a surfactant or a polyhydric alcohol as a carrier.

In one embodiment of the present invention, the fluid oil carrier may be a pharmaceutically acceptable carrier having non-aqueous or lipophilic properties. The fluid oil carrier may be substantially inert and not interact with the microspheres, and may be non-toxic and not have a negative effect on the patient. In addition, it may not dissolve the biocompatible polymer forming the microspheres. In addition, it may not dissolve the drug contained within the microspheres.

Additionally, microspheres using a fluid oil as a carrier can increase bioavailability compared to microspheres not using the fluid oil as a carrier.

The term “non-aqueous” above does not exclude the inclusion of trace amounts of residual water that do not adversely affect the solubility of the microspheres. The oil carrier may not dissolve the microspheres to such an extent that the stability of the microspheres is adversely affected or that the initial release control of the microspheres is lost. The oil carrier may also not swell the microspheres to such an extent that the stability of the microspheres is adversely affected.

The above fluid oil means a viscous liquid substance at ambient temperature or a slightly higher temperature, and has hydrophobicity (immiscible with water) and lipophilicity (literally miscible with other oils). As the oil carrier, particularly the fluid oil carrier comprising a fatty acid ester, at least one selected from the group consisting of coconut oil, palm oil, palm kernel oil, sesame oil, soybean oil, almond oil, rapeseed oil, corn oil, sunflower oil, peanut oil, olive oil, castor oil, safflower oil, cottonseed oil, and ethyl oleate can be used.

In one embodiment of the present invention, microspheres using a surfactant or a polyhydric alcohol as a carrier can increase bioavailability by 10% or more compared to microspheres not using the surfactant or the polyhydric alcohol as a carrier.

In one embodiment of the present invention, the surfactant may be at least one selected from Poloxamer 188, Poloxamer 407, Polysorbate 20, Polysorbate 60, Polysorbate 80, Polyoxyethylene oleyl ether, Polyoxyethylene cetyl ether, Polyoxyethylene stearyl ether, and Polyoxyethylene lauryl ether. More specifically, the surfactant may be, but is not limited to, Poloxamer 188, Poloxamer 407, polyoxyethylene oleyl ether, polyoxyethylene cetyl ether, polyoxyethylene stearyl ether or polyoxyethylene lauryl ether.

In one embodiment of the present invention, the polyhydric alcohol may be at least one selected from ethylene glycol, propylene glycol, polyethylene glycol, glycerol, erythritol, threitol, xylitol, ribitol, mannitol, sorbitol and maltitol. More specifically, the polyhydric alcohol may be ethylene glycol, propylene glycol, polyethylene glycol and glycerol, but is not limited to these examples.

In one embodiment of the present invention, the pharmaceutical composition may contain at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist contained in the microspheres in a concentration range of 0.01 mg to 2,000 mg, based on 1 mL of the first carrier. Here, the first carrier suspends at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist contained in the microspheres, and the pharmaceutical composition may be suspended including at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist in an amount of 0.01 mg to 2,000 mg, based on 1 mL of the first carrier.

More specifically, the concentration of the pharmaceutical composition is 0.01 mg or more, 0.05 mg or more, 0.1 mg or more, 0.15 mg or more, 0.2 mg or more, 0.25 mg or more, 0.3 mg or more, 0.35 mg or more, 0.4 mg or more, 0.45 mg or more, 0.5 mg or more, 0.55 mg or more, 0.6 mg or more, 0.65 mg or more, 0.7 mg or more, 0.75 mg or more, 0.8 mg or more, 0.85 mg or more, 0.9 mg or more, 0.95 mg or more, 1.0 mg or more, or 2,000 mg or less, 1,995 mg, of at least one selected from the group consisting of a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and/or a GLP-1/GIP/GCG receptor triple agonist contained in the microspheres, based on 1 mL of the first carrier. Below, 1,990 mg or less, 1,985 mg or less, 1,980 mg or less, 1,975 mg or less, 1,970 mg or less, 1,965 mg or less, 1,960 mg or less, 1,955 mg or less, 1,950 mg or less, 1,945 mg or less, 1,940 mg or less, 1,935 mg or less, 1,930 mg or less, 1,925 mg or less, 1,920 mg or less, 1,915 mg or less, 1,910 mg or less, 1,905 mg or less, 1,900 mg or less, but is not limited to these ranges.

In one embodiment of the present invention, at least one selected from the GLP-1 receptor agonist, the GIP/GLP-1 receptor agonist and the GLP-1/GIP/GCG receptor triple agonist may be selected from the group consisting of exenatide, liraglutide, lacxisenatide, albiglutide, dulaglutide, semaglutide, tirzepatide, cotadutide, taspoglutide, retatrutide and pharmaceutically acceptable salts thereof. More specifically, at least one selected from the GLP-1 receptor agonist, the GIP/GLP-1 receptor agonist and the GLP-1/GIP/GCG receptor triple agonist may be selected from the group consisting of semaglutide, tirzepatide, retatrutide and pharmaceutically acceptable salts thereof.

As the pharmaceutically acceptable salt, any salt commonly used in this field can be used without limitation. The term “pharmaceutically acceptable salt” of the present invention means any and all organic or inorganic addition salts of the compound, which have a relatively non-toxic and harmless effective effect on the patient, and whose side effects due to the salt do not reduce the beneficial effects of the active ingredient. Specific examples include, but are not limited to, acetate salts, benzoate salts, hydroxynaphthoate salts, napadiylate salts, and pamoate salts of each of the above drugs, and various forms of salts known in this field can all be included.

At least one selected from the above GLP-1 receptor agonists, GIP/GLP-1 receptor agonists and GLP-1/GIP/GCG receptor triple agonists may be manufactured in various forms, for example, may be amorphous or crystalline.

The microspheres according to one embodiment of the present invention mean microspheres manufactured using a biocompatible polymer in which the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist or GLP-1/GIP/GCG receptor triple agonist is encapsulated, and in the present specification, these are simply referred to as microspheres containing a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist or a GLP-1/GIP/GCG receptor triple agonist, a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist or a GLP-1/GIP/GCG receptor triple agonist microspheres, or microspheres.

In one embodiment of the present invention, a biocompatible polymer means a polymer that is guaranteed to be safe in vivo and does not cause high cytotoxicity or inflammatory responses when administered in vivo, and is also referred to simply as a polymer in this specification.

In one embodiment of the present invention, the microspheres may further include a biocompatible polymer. As the biocompatible polymer, any biocompatible polymer known in the art may be used without limitation, and specifically, for example, at least one selected from polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, copolymers of lactic acid and caprolactone, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acids, and copolymers of lactic acid and amino acids may be used.

In one embodiment of the present invention, the biocompatible polymer may be one having an intrinsic viscosity of 0.15 dL/g to 1.7 dL/g, 0.15 dL/g to 1.3 dL/g, or 0.15 dL/g to 1.2 dL/g.

The above intrinsic viscosity is measured at a concentration of 0.05 to 3% (w/v) in chloroform at 25°C using an Ubbelohde viscometer. When the above intrinsic viscosity is less than 0.15 dL/g, the molecular weight of the polymer is insufficient, making it difficult to exhibit a sustained-release effect of the drug, and when the intrinsic viscosity exceeds 1.7 dL/g, the release of the drug may be excessively delayed. In addition, when manufacturing microspheres using a polymer with high intrinsic viscosity, there is a problem that an excessive amount of manufacturing solvent must be used due to the high viscosity of the polymer, and it is difficult to manufacture reproducible microspheres.

Examples of commercially available polymers having the above properties include Evonik's Resomer series RG502H, RG503H, RG504H, RG502, RG503, RG504, RG653H, RG752H, RG752S, RG755S, RG750S, RG757S, RG858S, R202H, R203H, R205H, R202S, R203S, R205S, R206S, and R207S, Corbion's PDL 02A, PDL 02, PDL 04, PDL 05, PDLG 7502A, PDLG 7502, PDLG 7507, PDLG 5002A, PDLG 5002, PDLG 5004A, and Examples include PDLG 5004.

In one embodiment of the present invention, the microspheres may further comprise one or more release-controlling agents selected from butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, nonadecylic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, hydroxynaphthoic acid, napadiylic acid and pamoic acid.

In one embodiment of the present invention, at least one selected from the GLP-1 receptor agonist, the GIP/GLP-1 receptor agonist, and the GLP-1/GIP/GCG receptor triple agonist may be included in an amount of 3 to 50 wt% based on the total weight of the microparticles. More specifically, at least one selected from the GLP-1 receptor agonist, the GIP/GLP-1 receptor agonist and the GLP-1/GIP/GCG receptor triple agonist is present in an amount of 3.0 wt% or more, 3.5 wt% or more, 4.0 wt% or more, 4.5 wt% or more, 5.0 wt% or more, 5.5 wt% or more, 6.0 wt% or more, 6.5 wt% or more, 7.0 wt% or more, 7.5 wt% or more, 8.0 wt% or more, 8.5 wt% or more, 9.0 wt% or more, 9.5 wt% or more, 10.0 wt% or more, or 50 wt% or less, 49.5 wt% or less, 49.0 wt% or less, 48.5 wt% or less, 48.0 wt% or less, 47.5 wt% or less, 47.0 wt% or less, 46.5 wt% based on the total weight of the microspheres. Below, 46.0 wt% or less, 45.5 wt% or less, 45.0 wt% or less, 44.5 wt% or less, 44.0 wt% or less, 43.5 wt% or less, 43.0 wt% or less, 42.5 wt% or less, 42.0 wt% or less, 41.5 wt% or less, 41.0 wt% or less, 40.5 wt% or less, 40.0 wt% or less.

If at least one selected from the above GLP-1 receptor agonist, GIP/GLP-1 receptor agonist, and GLP-1/GIP/GCG receptor triple agonist is contained in less than 3 wt%, it is not preferable because long-term efficacy cannot be exhibited with a single administration. In addition, if at least one selected from the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist, and GLP-1/GIP/GCG receptor triple agonist is contained in more than 50 wt%, the initial release amount of the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist, or GLP-1/GIP/GCG receptor triple agonist in the body environment may be too high, which may cause a problem in that the blood concentration of the drug rapidly increases.

In addition, the content range of the biocompatible polymer included in the microparticles may be 50 to 97 wt% based on the total weight of the microparticles. More specifically, the content of the biocompatible polymer may be 50 wt% or more, 55 wt% or more, 60 wt% or more, or 97 wt% or less, 96.0 wt% or less, 95.0 wt% or less, 94.0 wt% or less, 93.0 wt% or less, 92.0 wt% or less, 91.0 wt% or less, or 90.0 wt% or less based on the total weight of the microparticles, but is not limited to these ranges.

When the biocompatible polymer exceeds 97 wt% in the above, the drug release may become too slow to secure a constant blood drug profile, and when the biocompatible polymer is included in an amount of less than 50 wt% based on the total weight of the microparticles, the distribution of the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist, or GLP-1/GIP/GCG receptor triple agonist may relatively increase, resulting in problems such as initial excessive release or inability to maintain the efficacy for a desired period.

In one embodiment of the present invention, the microspheres are mixed with at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist, and the biocompatible polymer, for example, at a ratio of 1:1 to 50, 1:1.5 to 50, 1:1.6 to 50, 1:1.7 to 50, 1:1 to 47, 1:1.5 to 47, 1:1.6 to 47 1:1.7 to 47, 1:1 to 45, 1:1.5 to 45, 1:1.6 to 45, 1:1.7 to 45, 1:1 to 42, 1:1.5 to 42, 1:1.6 to 42, 1:1.7 to 42, 1:1 to 40, 1:1.5 to 40, 1:1.6 to 40, It can be included in a weight ratio of 1:1.7~40, 1:1~38, 1:1.5~38, 1:1.6~38, 1:1.7~38, 1:1~35, 1:1.5~35, 1:1.6~35, or 1: 1.7~35.

At this time, if the weight ratio of the biocompatible polymer to at least one selected from the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist, and GLP-1/GIP/GCG receptor triple agonist is less than 1.0, it is undesirable because fatal side effects may occur due to the initial drug excessive release phenomenon, and if the weight ratio exceeds 50, the drug dissolution may be too slow, making it difficult to secure a constant blood drug profile.

In one embodiment of the present invention, the microspheres may have a release rate of at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist of 60 wt% or less, 55 wt% or less, or 50 wt% or less within 7 days.

Additionally, the microspheres may have a release rate of at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist of 25 wt% or less, 23 wt% or less, or 20 wt% or less within 3 days.

In particular, the microspheres of the present invention may have a drug release rate of at least one drug selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist of 10 wt% or less, 8 wt% or less, or 5 wt% or less during the first day of administration.

In addition, the microspheres of the present invention may continuously release the drug for 10 days or more, 20 days or more, 1 month or more, 2 months or more, 3 months or more, 4 months or more, 5 months or more, or 6 months or more.

In one embodiment of the present invention, the microspheres can be manufactured by various microsphere manufacturing methods known in the art (e.g., O/W type, O/O type or W/O/W type solvent evaporation method or solvent extraction method, microsphere manufacturing method by spray drying, microsphere manufacturing method by phase separation, etc.).

In one embodiment of the present invention, the microspheres may be prepared by a solvent evaporation or extraction method using an emulsion, more preferably, an O/W (oil-in-water) type emulsion including a biocompatible polymer; at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist; and a dispersion solvent; and agglomerating the emulsion into microspheres.

In order to manufacture microspheres by preparing the above O/W type emulsion and coagulating it into polymer microparticles, first, an O/W type emulsion including a biocompatible polymer; at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist; and a dispersion solvent is prepared.

The above O/W type emulsion can be manufactured using a conventional method known in the art, and more specifically, it can be manufactured by adding a dispersion phase including at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist and a biocompatible polymer to a dispersion solvent.

Microspheres containing at least one selected from these GLP-1 receptor agonists, GIP/GLP-1 receptor agonists and GLP-1/GIP/GCG receptor triple agonists can be prepared by coagulating the emulsion into microspheres by solvent extraction and/or solvent evaporation, or by coagulation by ammonolysis with addition of ammonia or by hydrolysis with addition of acid or base. A water-insoluble organic solvent which is converted into a water-soluble solvent by ammonolysis or hydrolysis reaction can be additionally included during the preparation of the emulsion.

In the case of the solvent evaporation method, the present invention is not limited thereto, but for example, the method described in U.S. Patent Nos. 5,271,945, 5,985,309 and 6,471,996, etc., i.e., dissolving a polymer compound in an organic solvent, dispersing or dissolving a drug in the organic solvent, emulsifying in a dispersion medium such as water to prepare an O/W type emulsion, and then diffusing the organic solvent in the emulsion into the dispersion medium and evaporating through the air/water interface, thereby forming polymer microspheres containing at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and a GLP-1/GIP/GCG receptor triple agonist.

The above solvent extraction method includes a conventional solvent extraction method used for the production of microspheres containing at least one selected from among GLP-1 receptor agonists, GIP/GLP-1 receptor agonists and GLP-1/GIP/GCG receptor triple agonists, such as effectively extracting the organic solvent in the emulsion droplets using a large amount of solubilizing solvent.

As a method of simultaneously applying the solvent evaporation method and the solvent extraction method, for example, methods described in U.S. Patent Nos. 4,389,840, 4,530,840, 6,368,632, 6,544,559, and 6,572,894 can be applied.

The coagulation by the above ammonolysis process refers to a method of coagulating fine particles by adding ammonia to an O/W type emulsion containing a water-insoluble organic solvent to induce ammonolysis, thereby converting the water-insoluble organic solvent into a water-soluble solvent, as described in, for example, Korean Patent No. 918092.

The coagulation by the above hydrolysis process refers to a method of coagulating fine particles by adding a base such as NaOH, LiOH, or KOH, or an acid solution such as HCl or H2SO4 to an O/W type emulsion containing a water-insoluble organic solvent, thereby inducing hydrolysis, which is a type of hydrolysis reaction of ester, and converting the water-insoluble organic solvent into a water-soluble solvent, as described in, for example, Korean Patent Application Nos. 2009-109809 and 2010-70407.

In one embodiment of the present invention, the microspheres may be W1/O/W2 (water-in-oil-in-water) type microspheres prepared by preparing a W1/O (water-in-oil) type emulsion including an inner water phase (W1) in which at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist is dispersed or dissolved in an aqueous solvent; and an oil phase (O) in which a biocompatible polymer is dissolved in a non-aqueous solvent; and dispersing the same in an outer water phase (W2). In this case, the microspheres may be prepared by a double emulsification evaporation method.

In order to manufacture microspheres by preparing the above W/O/W type emulsion and coagulating it into polymer microparticles, first, a W/O/W type emulsion containing at least one selected from a biocompatible polymer, a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist, and a dispersion solvent is prepared.

At this time, the production of the W/O/W type emulsion can be performed using a conventional method known in the art, and more specifically, it can be produced by adding a dispersion phase including at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist and a biocompatible polymer to a dispersion solvent.

Microspheres containing at least one selected from these GLP-1 receptor agonists, GIP/GLP-1 receptor agonists and GLP-1/GIP/GCG receptor triple agonists can be prepared by coagulating the emulsion into microspheres by solvent extraction and/or solvent evaporation, or by coagulation by ammonolysis with addition of ammonia or by hydrolysis with addition of acid or base. A water-insoluble organic solvent which is converted into a water-soluble solvent by ammonolysis or hydrolysis reaction can be additionally included during the preparation of the emulsion.

The solvent evaporation method, solvent extraction method, coagulation by ammonolysis process, and coagulation by hydrolysis process can be performed in the same manner as described above.

In one embodiment of the present invention, the pharmaceutical composition may further comprise one or more pharmaceutically acceptable additives. The additives may be, for example, one or more selected from the group consisting of a buffer, an isotonic agent, a preservative, and a pH adjusting agent.

The above additive is distinct from the first carrier, and the pharmaceutical composition according to the present invention can be formulated in a suitable form together with the microparticles and a pharmaceutically acceptable carrier.

For example, phosphate buffer, TRIS, citrate, etc. can be used as the above buffer, but are not limited thereto, and buffers known in the art can be used without limitation. The phosphate buffer can include sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium phosphate, etc. The buffer can be included at a concentration of, for example, 0.01 to 50 mM.

Examples of the isostatic agent that can be used include, but are not limited to, propylene glycol, sodium chloride, mannitol, sucrose, sorbitol, trehalose, lactose, etc., and isostatic agents known in the art can be used without limitation. The isostatic agent can be included in the liquid formulation at a concentration of 8 mg/ml to 50 mg/ml, for example, a concentration of 8 mg/ml to 30 mg/ml.

For example, phenol and the like can be used as the above preservative, but are not limited thereto, and any preservative known in this field can be used without limitation.

Examples of pH regulators that can be used include, but are not limited to, HCl, NaOH, or acetate, and any pH regulator known in the art can be used without limitation.

The pharmaceutical composition of the present invention can be formulated into various forms of preparations, for example, can be formulated into parenteral administration preparations. The parenteral administration preparations can be, for example, injections, creams, lotions, ointments for external use, oils, moisturizers, gels, aerosols, patches, nasal inhalers, etc., but are not limited thereto.

The above parenteral administration formulation may further include, in addition to the microparticles, a thickening agent, a stabilizer, an isotonic agent, a pH regulator, a surfactant, an excipient and/or a carrier.

The above usable isotonic agents include water-soluble excipients or sugars such as mannitol, sucrose, sorbitol, trehalose, lactose, and sodium chloride.

Examples of the thickeners include sodium carmellose, sodium carboxymethyl cellulose, and povidone.

Examples of the above surfactants include polyvinyl alcohol, etc. In addition, examples of buffering agents include sodium dihydrogen phosphate, anhydrous citric acid, sodium hydroxide, sodium chloride, etc.

The above parenteral administration formulation is not limited to the forms exemplified above and may be formulated in a manner known in the art.

In one embodiment of the present invention, the pharmaceutical composition may be an injection.

In one embodiment of the present invention, the pharmaceutical composition may be an injectable preparation for subcutaneous or intramuscular injection.

In one embodiment of the present invention, the pharmaceutical composition may be administered to a subject at intervals of one month to one year.

In one embodiment of the present invention, the pharmaceutical composition can be used to prevent or treat diabetes, preservation of beta-cell function, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, or degenerative neurological diseases such as Alzheimer's disease and Parkinson's disease.

The pharmaceutical composition of the present invention can be administered in a therapeutically effective amount of a GLP 1 receptor agonist, a GIP/GLP-1 receptor agonist or a GLP-1/GIP/GCG receptor triple agonist, for example, an effective amount for treating diabetes, specifically type 2 diabetes, preservation of beta-cell function, hypertension, hyperlipidemia, obesity, nonalcoholic steatohepatitis, or degenerative neurological diseases such as Alzheimer's disease and Parkinson's disease. The therapeutically effective amount of a GLP 1 receptor agonist, a GIP/GLP-1 receptor agonist or a GLP-1/GIP/GCG receptor triple agonist can be assessed by a physician.

The pharmaceutical composition of the present invention may be administered once every two weeks, once every three weeks, once every month, once every three months, once every six months, or once every year, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered, for example, by subcutaneous injection. The subcutaneous injection may be performed, for example, using a prefilled syringe, for example, a pen-syringe, which may be a disposable syringe containing a single dose, or a metered dose syringe (syringe with dose metering device) that injects only a single dose at a time.

The pharmaceutical composition of the present invention exhibits sufficient efficacy as at least one of a GLP 1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist during the treatment period, and thus can be usefully used for the prevention or treatment of diabetes, specifically type 2 diabetes, preservation of beta-cell function, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, or degenerative neurological diseases such as Alzheimer's disease and Parkinson's disease.

The pharmaceutical composition of the present invention can be manufactured, transported and stored in a state in which microspheres containing at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and a GLP-1/GIP/GCG receptor triple agonist are suspended in a fluid oil carrier containing a fatty acid ester.

In addition, the pharmaceutical composition of the present invention can be manufactured, transported and stored in a state in which microspheres including at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and a GLP-1/GIP/GCG receptor triple agonist are suspended in a carrier containing a surfactant or a polyhydric alcohol.

The pharmaceutical composition of the present invention may be provided in a sealed container. The sealed container may be, for example, a vial (disposable or multi-use), a syringe, an injection pen (for example, disposable or multi-use), etc.

The pharmaceutical composition of the present invention provides long-term storage stability. For example, the pharmaceutical composition of the present invention can be stored in a stable state for 6 months or more, 1 year or more, 1 year and 6 months or more, or 2 years or more. The “stable state” means a state in which the activity of the drug can be maintained by at least 90% or more, 95% or more, or 99% or more compared to the activity in the initial formulation (i.e., 100%). In particular, it means that the particle size and/or shape of the microspheres are maintained, and thus the drug activity is maintained within the above range.

The microparticles included in the pharmaceutical composition of the present invention can be prepared using a method known in the art, for example, a “solvent extraction and evaporation method.”

Specifically, it can be manufactured by a method comprising the following steps:

(a) a step of preparing a solution (dispersed phase) containing at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist and a polymer by dissolving a biocompatible polymer in an organic solvent;

(b) a step of preparing an emulsion by adding at least one selected from the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and GLP-1/GIP/GCG receptor triple agonist prepared in the above step (a) and a polymer-containing solution to an aqueous solution phase (continuous phase);

(c) a step of forming microspheres by continuously extracting and evaporating an organic solvent from the dispersed phase of the emulsion state prepared in step (b); and

(d) a step of recovering microspheres from the continuous phase of step (c) and preparing microspheres containing at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist.

In the above manufacturing method, at least one selected from among the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and GLP-1/GIP/GCG receptor triple agonist and the biocompatible polymer, etc. have been described above, and therefore, a description thereof is omitted here.

In step (a) above, at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist can be first dissolved in an organic solvent and then mixed with a biocompatible polymer solution.

In the above step (a), an organic solvent that is miscible with water or an organic solvent that is not miscible with water may be used. Alternatively, a mixed solvent of an organic solvent that is miscible with water and an organic solvent that is not miscible with water may be used.

It is preferable to use the organic solvent having a property of being immiscible with water in an amount of 50% (v/v) or more of the total solvent. When the property of the organic solvent not being miscible with water is utilized, an emulsion can be efficiently formed by homogeneously mixing the dispersed phase in the continuous phase.

The type of organic solvent that dissolves at least one selected from the above GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and GLP-1/GIP/GCG receptor triple agonist and the biocompatible polymer is not particularly limited, but preferably, one or two or more mixed solvents selected from the group consisting of dichloromethane, chloroform, ethyl acetate, methyl ethyl ketone, acetone, acetonitrile, dimethyl sulfoxide, dimethyl formamide, N-methyl pyrrolidone, acetic acid, methyl alcohol, ethyl alcohol, propyl alcohol and benzyl alcohol can be used.

In the step (b) above, the step of adding at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist and a polymer-containing solution (dispersed phase) to an aqueous solution phase (continuous phase) to prepare an emulsion can be performed using a homogenizer such as a high-speed stirrer, an in-line mixer, a membrane emulsion method, a microfluidic emulsion method, an ultrasonic mixer, or a static mixer.

In the step (b), the continuous phase may further include a surfactant. The type of the surfactant is not particularly limited, and any surfactant that can help the polymer solution form a stable droplet dispersion phase in the continuous phase, as well as at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist, may be used. As the surfactant, a hydrophilic polymer selected from the group consisting of methylcellulose, polyvinylpyrrolidone, carboxymethylcellulose, lecithin, gelatin, polyvinyl alcohol, polyoxyethylene-polyoxypropylene block copolymers, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene castor oil derivatives, and mixtures thereof may be used, and polyvinyl alcohol is particularly preferable.

The surfactant may be added to the continuous phase at a concentration of 0.1 to 3% (w/v), more preferably 0.3 to 2% (w/v). When the concentration of the surfactant is less than 0.1%, a dispersed phase or emulsion in the form of droplets may not be formed in the continuous phase, and when the concentration of the surfactant exceeds 3%, it may be difficult to remove the surfactant after the microspheres are formed.

Water can be used as the aqueous solution (continuous phase) of the above step (b), and the pH of the aqueous solution can be 4.0 to 10, but is not limited thereto. NaCl, which acts as an osmotic regulator, can be used to control the initial release.

In the step (c) above, if the emulsion is maintained or stirred at a temperature below the boiling point of the organic solvent for a certain period of time, for example, 2 hours to 48 hours, the organic solvent can be extracted from the dispersed phase into the continuous phase. A portion of the organic solvent extracted into the continuous phase can be evaporated from the surface. At this time, the dispersed phase in the form of droplets can be solidified to form microspheres.

In order to efficiently remove the organic solvent in the step (c) above, the continuous phase can be heated for a certain period of time. The heating temperature is not limited and can be appropriately adjusted by a person skilled in the art depending on the organic solvent used.

The method of recovering the microparticles in the step (d) above can be performed using various known techniques, for example, methods such as filtration or centrifugation can be used.

Between the above steps (c) and (d), it is also possible to remove the remaining surfactant through filtration and washing, and then filter again to recover the microspheres. The washing step for removing the remaining surfactant can be typically performed using water, and the washing step can be repeated several times.

The microparticles obtained after the above step (d) can be dried using a conventional drying method to finally obtain dried microparticles.

The microparticles of the present invention can also be manufactured by the following method.

(1) A step of preparing an inner phase (W1) by dissolving or dispersing at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist in a solvent;

(2) A step of preparing an oil phase (O) by dissolving a biocompatible polymer in one or more solvents;

(3) A step of dispersing an internal phase (W1) in the oil phase (O) to produce a water-in-oil (W1/O) type emulsion;

(4) a step of dispersing the water-in-oil (W1/O) type emulsion into an external continuous phase (W2) to produce a water-in-oil (W1/O/W2) type emulsion to form microspheres; and

(5) A method for manufacturing microparticles is provided, including a step of removing the solvent.

In the method for manufacturing the microspheres of the present invention, all of the contents of the microspheres and the method for manufacturing the microspheres described above that are compatible with the manufacturing method (type of biocompatible polymer, intrinsic viscosity thereof, etc.) can be applied. Therefore, description of overlapping contents is omitted below.

The above step (1) may be a step of preparing a water phase (W1) by dissolving at least one selected from, for example, a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist in distilled water.

In the above step (2), the type of solvent is not particularly limited, and for example, dimethyl sulfoxide, dichloromethane, or a mixed solvent thereof can be used. In particular, dichloromethane can be preferably used.

The above step (3) is a step of producing a water-in-oil (W1/O) type emulsion by dispersing the inner phase (W1) produced in step (1) into the oil phase (O) produced in step (2).

In the above step (3), the weight ratio of the oil phase (O) to the internal phase (W1) may be 1:10 or less.

The above step (4) is a step of solidifying microspheres by dispersing the water-in-oil (W1/O) type dispersed phase prepared in the above step (3) into an external continuous phase (W2) to prepare a water-in-oil-in-water (W1/O/W2) type emulsion solution.

In the above step (4), the external continuous phase (W2) may contain a hydrophilic polymer as a surfactant, and the type thereof is not particularly limited, and any one or more selected from among a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist, and a dispersed phase including a biocompatible polymer may be used as long as it can help form a stable droplet dispersion phase within the external continuous phase.

In the above step (4), the external continuous phase may be a hydrophilic polymer aqueous solution of 0.1 to 6% (w/v), preferably 0.1 to 4% (w/v).

In the step (4) above, at least one selected from the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and GLP-1/GIP/GCG receptor triple agonist manufactured in the step (3) above and a dispersed phase (W1/O) including a biocompatible polymer are added to the external continuous phase including the hydrophilic polymer by drop-by-drop or by using an in-line mixer, and stirred vigorously to manufacture a water-in-oil-in-water (W1/O/W2) type emulsion solution.

Afterwards, in the step (5) above, the solvent is removed, and after conventional filtration and washing, the desired microspheres can be obtained. That is, a step of washing the obtained microspheres using an organic solvent such as ethanol may be included to enhance the initial release inhibition effect, if necessary.

Hereinafter, the present invention will be described in detail by way of examples in order to specifically explain the present invention. However, the examples according to the present invention may be modified in various different forms, and the scope of the present invention should not be construed as being limited to the examples described below. The examples of the present invention are provided in order to more completely explain the present invention to a person having average knowledge in the art.

Manufacturing Example 1: Manufacturing of Semaglutide Microspheres

<Continuous Manufacturing>

900 ml of distilled water was added to the reactor, and 4.5 g of polyvinyl alcohol (PVA) and 4.5 g of sodium chloride (NaCl) were weighed and added. A 0.5% (w/v) polyvinyl alcohol aqueous solution was prepared by stirring at 1,000 rpm using a homogenizer (Overhead (IKA)).

<Preparation of Semaglutide Solution>

0.32 g of semaglutide and 2.4 ml of distilled water were weighed and added to a 20 ml vial, and stirred at 150 rpm to dissolve. The prepared solution was stored in the refrigerator.

<Preparation of PLGA solution>

2.88 g of PLGA (inherent viscosity 0.15 dL/g to 1.7 dL) was dissolved in 9.6 ml of dichloromethane and stirred at 300 rpm.

<Emulsification>

The PLGA solution prepared above was taken up with a syringe, added to the semaglutide solution, and stirred at 20,000 rpm for 1 minute using a homogenizer (IKA).

<Recovery of microspheres>

After the above emulsification process was completed, the solution was immediately taken into a syringe and placed into a 1 L reactor containing the continuous phase in advance, and stirred for 60 seconds using a homogenizer (Kinematica). The microspheres were then solidified for 20 hours. After solidification was complete, the solution was filtered through a 5 μm filter to obtain microspheres (W/O/W type). The obtained microspheres were then freeze-dried in a freeze-dryer for more than 20 hours.

Manufacturing Example 2: Manufacturing of Semaglutide Microspheres

<Continuous Manufacturing>

900 ml of distilled water was added to the reactor, and 4.5 g of polyvinyl alcohol (PVA) and 4.5 g of sodium chloride (NaCl) were weighed and added. A 0.5% (w/v) polyvinyl alcohol aqueous solution was prepared by stirring at 1,000 rpm using a homogenizer (Overhead (IKA)).

<Preparation of Semaglutide Solution>

0.48 g of semaglutide and 2.4 ml of distilled water were weighed and added to a 20 ml vial, and stirred at 150 rpm to dissolve.

<Preparation of PLGA solution>

1.44 g of PLGA (inherent viscosity 0.15 dL/g to 1.7 dL/g) was dissolved in 4.8 ml of dichloromethane and stirred at 300 rpm.

<Emulsification>

The PLGA solution prepared above was taken up with a syringe, added to the semaglutide solution, and stirred at 20,000 rpm for 1 minute using a homogenizer (IKA).

<Recovery of microspheres>

After the above emulsification process was completed, the solution was immediately taken into a syringe and placed into a 1 L reactor containing the continuous phase in advance, and stirred for 60 seconds using a homogenizer (Kinematica). The microspheres were then solidified for 20 hours. After solidification was complete, the solution was filtered through a 5 μm filter to obtain microspheres (W/O/W type). The obtained microspheres were then freeze-dried in a freeze-dryer for more than 20 hours.

Example 1: Preparation of a liquid formulation injection containing semaglutide microspheres and MCT oil

Semaglutide microspheres of Manufacturing Example 1 corresponding to 3 mg/kg of semaglutide were weighed and placed in a vial, and MCT oil was added thereto to suspend the mixture to prepare a semaglutide microsphere injection.

Example 2: Preparation of liquid formulation injection containing semaglutide microspheres and castor oil

A semaglutide microsphere injection was prepared in the same manner as in Example 1, except that MCT oil was replaced with Castor oil.

Example 3: Preparation of liquid formulation injection containing semaglutide microspheres and cottonseed oil

A semaglutide microsphere injection was prepared in the same manner as in Example 1, except that MCT oil was replaced with cottonseed oil.

Example 4: Preparation of liquid formulation injection containing semaglutide microspheres and sesame oil

A semaglutide microsphere injection was prepared in the same manner as in Example 1, except that MCT oil was replaced with sesame oil.

Example 5: Preparation of liquid formulation injection containing semaglutide microspheres and Polyoxyl 35 castor oil

A semaglutide microsphere injection was prepared in the same manner as in Example 1, except that MCT oil was replaced with Polyoxyl 35 castor oil.

Example 6: Preparation of a liquid formulation injection comprising semaglutide microspheres and propylene glycol liquid carrier

The microspheres manufactured in the above Manufacturing Example 2 were weighed so that the dosage of semaglutide would be 1 mg/kg, and then propylene glycol as a carrier was added and mixed to manufacture a semaglutide microsphere injection.

Example 7: Preparation of liquid formulation injection containing semaglutide microspheres and MCT oil

A semaglutide microsphere injection was prepared in the same manner as in Example 6, except that propylene glycol was replaced with MCT oil.

Comparative Example 1: Preparation of a liquid formulation injection containing semaglutide microspheres and a diluent

A semaglutide microsphere injection was prepared in the same manner as in Example 1, except that MCT oil was replaced with a diluent containing anhydrous citric acid, sodium phosphate monohydrate, sodium carboxymethylcellulose, polysorbate 20, sodium chloride, and sodium hydroxide.

Comparative Example 2: Preparation of a liquid formulation injection containing semaglutide microspheres and a diluent

A semaglutide microsphere injection was prepared in the same manner as in Example 6, except that propylene glycol was replaced with a diluent containing anhydrous citric acid, sodium phosphate monohydrate, sodium carboxymethylcellulose, polysorbate 20, sodium chloride, and sodium hydroxide.

Manufacturing Example 3: Manufacturing of tirzepatide microspheres

<Continuous Manufacturing>

900 ml of distilled water was added to the reactor, and 4.5 g of polyvinyl alcohol (PVA) and 4.5 g of sodium chloride (NaCl) were weighed and added. A 0.5% (w/v) polyvinyl alcohol aqueous solution was prepared by stirring at 1,000 rpm using a homogenizer (Overhead (IKA)).

<Preparation of Tirzepatide Solution>

0.3 g of tirzepatide and 1.0 ml of distilled water were weighed and added to a 20 ml vial, and stirred at 150 rpm to dissolve.

<Preparation of PLGA solution>

1.2 g of PLGA (inherent viscosity 0.15 dL/g to 1.7 dL/g) was dissolved in 3 ml of dichloromethane by stirring at 300 rpm. The prepared solution was stored in the refrigerator.

<Emulsification>

The PLGA solution prepared above was taken up with a syringe, added to the tirzepatide solution, and stirred at 20,000 rpm for 1 minute using a homogenizer (IKA).

Manufacturing Example 4: Manufacturing of tirzepatide microspheres

<Continuous Manufacturing>

900 ml of distilled water was added to the reactor, and 4.5 g of polyvinyl alcohol (PVA) and 4.5 g of sodium chloride (NaCl) were weighed and added. A 0.5% (w/v) polyvinyl alcohol (PVA) aqueous solution was prepared by stirring at 1,000 rpm using a homogenizer (Overhead (IKA)).

<Preparation of Tirzepatide Solution>

In a 20 mL vial, 0.386 g of tirzepatide and 1.5 mL of distilled water were weighed and placed, stirred at 150 rpm to dissolve, and then 0.037 mL of propylene glycol was added and stirred to dissolve.

<Preparation of PLGA solution>

0.9 g of PLGA (inherent viscosity 0.15 dL/g to 1.7 dL/g) was dissolved in 3 ml of dichloromethane and stirred at 300 rpm.

<Emulsification>

The PLGA solution prepared above was taken up with a syringe, added to the tirzepatide solution, and stirred at 20,000 rpm for 1 minute using a homogenizer (IKA).

<Recovery of microspheres>

After the above emulsification process was completed, the solution was immediately taken into a syringe and placed into a 1 L reactor containing the continuous phase in advance, and stirred for 60 seconds using a homogenizer (Kinematica). The microspheres were then solidified for 20 hours. After solidification was complete, the solution was filtered through a 5 μm filter to obtain microspheres (W/O/W type). The obtained microspheres were then freeze-dried in a freeze-dryer for more than 20 hours.

Example 8: Preparation of liquid formulation injection containing tirzepatide microspheres and MCT oil

The tirzepatide microspheres of Manufacturing Example 3 corresponding to 3 mg/kg of tirzepatide were weighed and placed in a vial, and MCT oil was added thereto to suspend the mixture to prepare a tirzepatide microsphere injection.

Example 9: Preparation of a liquid formulation injection containing tirzepatide microspheres and castor oil

A tirzepatide microsphere injection was prepared in the same manner as in Example 8, except that MCT oil was replaced with Castor oil.

Example 10: Preparation of liquid formulation injection containing tirzepatide microspheres and cottonseed oil

A tirzepatide microsphere injection was prepared in the same manner as in Example 8, except that MCT oil was replaced with cottonseed oil.

Example 11: Preparation of a liquid formulation injection comprising tirzepatide microspheres and sesame oil

A tirzepatide microsphere injection was prepared in the same manner as in Example 8, except that MCT oil was replaced with sesame oil.

Example 12: Preparation of liquid formulation injection containing tirzepatide microspheres and Polyoxyl 35 castor oil

A tirzepatide microsphere injection was prepared in the same manner as in Example 8, except that MCT oil was replaced with Polyoxyl 35 castor oil.

Example 13 Preparation of a liquid formulation injection comprising tirzepatide microparticles and propylene glycol liquid carrier

The microspheres manufactured in the above Manufacturing Example 4 were weighed so that the dosage of tirzepatide would be 1 mg/kg, then propylene glycol as a carrier was added, and these were mixed to manufacture a tirzepatide microsphere injection.

Comparative Example 3: Preparation of a liquid formulation injection containing tirzepatide microspheres and a diluent

A tirzepatide microsphere injection was prepared in the same manner as in Example 8, except that MCT oil was replaced with a diluent including anhydrous citric acid, sodium phosphate monohydrate, sodium carboxymethylcellulose, polysorbate 20, sodium chloride, and sodium hydroxide.

Comparative Example 4: Preparation of liquid injection formulation of tirzepatide microspheres

A tirzepatide microsphere injection was prepared in the same manner as in Example 13, except that propylene glycol was replaced with a diluent including anhydrous citric acid, sodium phosphate monohydrate, sodium carboxymethylcellulose, polysorbate 20, sodium chloride, and sodium hydroxide.

Experimental Example 1: In-vivo pharmacokinetics test using beagles

The above semaglutide microparticle injection formulation (Comparative Example 2) and the injection formulation containing semaglutide microparticles and propylene glycol (Example 6) were administered to beagles, and then the blood semaglutide concentration was measured.

Specifically, each sample was injected subcutaneously into a beagle using a 23-gauge syringe, and blood was collected at designated times, and the blood concentration of semaglutide was measured using LC-MS/MS. The change in the blood concentration of semaglutide is shown in Figure 1.

Experimental Example 2: In-vivo pharmacokinetics test using beagles

The above tirzepatide microparticle injection formulation (Comparative Example 4) and the injection formulation containing tirzepatide microparticles and propylene glycol (Example 13) were administered to beagles, and then the blood semaglutide concentration was measured.

Specifically, each sample was injected subcutaneously into a beagle using a 23-gauge syringe, blood was collected at designated times, and the blood concentration of semaglutide was measured using LC-MS/MS.

Claims (15)

Microspheres comprising at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist and a GLP-1/GIP/GCG receptor triple agonist; and
Containing at least one selected from a first carrier and a water-soluble liquid,
A pharmaceutical composition wherein the first carrier comprises a fluid oil containing a fatty acid ester, a surfactant or a polyhydric alcohol. In the first paragraph,
A pharmaceutical composition wherein the fluid oil is at least one selected from coconut oil, palm oil, palm kernel oil, sesame oil, soybean oil, almond oil, rapeseed oil, corn oil, sunflower oil, peanut oil, olive oil, castor oil, safflower oil, cottonseed oil, and ethyl oleate. In the first paragraph,
A pharmaceutical composition, wherein the surfactant is at least one selected from Poloxamer 188, Poloxamer 407, Polysorbate 20, Polysorbate 60, Polysorbate 80, Polyoxyethylene oleyl ether, Polyoxyethylene cetyl ether, Polyoxyethylene stearyl ether, and Polyoxyethylene lauryl ether. In the first paragraph,
A pharmaceutical composition, wherein the polyhydric alcohol is at least one selected from ethylene glycol, propylene glycol, polyethylene glycol, glycerol, erythritol, threitol, xylitol, ribitol, mannitol, sorbitol, and maltitol. In paragraph 4,
A pharmaceutical composition, wherein the polyhydric alcohol is ethylene glycol, propylene glycol, polyethylene glycol or glycerol. In the first paragraph,
The pharmaceutical composition is a pharmaceutical composition comprising at least one selected from a GLP-1 receptor agonist, a GIP/GLP-1 receptor agonist, and a GLP-1/GIP/GCG receptor triple agonist contained in microspheres in a concentration range of 0.01 mg to 2,000 mg based on 1 mL of the first carrier. In the first paragraph,
A pharmaceutical composition, wherein the GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and/or GLP-1/GIP/GCG receptor triple agonist is at least one selected from exenatide, liraglutide, laxenatide, albiglutide, dulaglutide, semaglutide, tirzepatide, cotadutide, taspoglutide, retatrutide and pharmaceutically acceptable salts thereof. In the first paragraph,
A pharmaceutical composition, wherein at least one selected from the above GLP-1 receptor agonist, GIP/GLP-1 receptor agonist and GLP-1/GIP/GCG receptor triple agonist is contained in an amount of 3 wt% to 50 wt% based on the total weight of microspheres. In the first paragraph,
A pharmaceutical composition wherein the microparticles further comprise a biocompatible polymer. In Article 9,
A pharmaceutical composition wherein the biocompatible polymer is at least one selected from polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, copolymer of lactic acid and caprolactone, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid, and copolymer of lactic acid and amino acid. In Article 9,
A pharmaceutical composition, wherein the biocompatible polymer is contained in an amount of 50 to 97 wt% based on the total weight of the microparticles. In the first paragraph,
A pharmaceutical composition further comprising one or more additives selected from a buffer, an isotonic agent, a preservative, and a pH adjusting agent. In Article 9,
A pharmaceutical composition, wherein the microspheres contain at least one selected from the GLP-1 receptor agonist, the GIP/GLP-1 receptor agonist, and the GLP-1/GIP/GCG receptor triple agonist, and the biocompatible polymer in a weight ratio of 1:1 to 50. In the first paragraph,
A pharmaceutical composition, wherein the microparticles remain stable for more than 6 months. In the first paragraph,
A pharmaceutical composition, wherein the above pharmaceutical composition is an injection.