Pharmaceutical compositions, their uses, and methods for improving the storage stability of pharmaceutical compositions.

Abstract

This application provides embodiments of a stable pharmaceutical composition, a method for improving the storage stability of the pharmaceutical composition, and uses of the stable pharmaceutical composition. The pharmaceutical composition comprises a first active ingredient containing a GLP-1 receptor agonist and a surfactant. In one embodiment, the surfactant contains an alkyl glycoside. In one embodiment, one or more alkyl glycosides (not limited to, for example, DDM) inhibit the formation of GLP-1 fibrillation in the pharmaceutical composition containing a GLP-1 receptor agonist such as GLP-1 or an analog or a derivative of a GLP-1 analog, thereby improving physical stability.

Description

[Technical Field]

[0001] [Claiming priority] This application claims priority under Chinese Patent Application No. 2023105246295, filed on 10 May 2023, the entire contents of which are incorporated herein by reference.

[0002] This disclosure belongs to the field of biotechnology / pharmaceuticals. More specifically, this disclosure relates to a pharmaceutical composition comprising a GLP-1 receptor agonist, a method for improving the storage stability of the pharmaceutical composition, and applications for manufacturing pharmaceuticals using the pharmaceutical composition. [Background technology]

[0003] There are approximately 120 million people worldwide with diabetes. Exogenous insulin is the primary treatment for diabetes. Insulin is the only hormone in the body that lowers blood glucose levels while promoting the synthesis of glycogen, lipids, and proteins. Insulin glargine is an analog of human insulin and can be obtained through recombinant DNA technology. When insulin glargine is injected subcutaneously, a small amount of insulin glargine is continuously released from a fine precipitate formed by the neutralization of an acidic solution. This results in a predictable, long-acting, smooth, and peakless plasma concentration / time profile.

[0004] Glucagon-like peptide 1 (hereinafter referred to as GLP-1) is an incretin hormone with the following physiological effects: it promotes insulin secretion in a glucose concentration-dependent manner, thereby reducing the risk of hypoglycemia; it promotes insulin synthesis in the body; it promotes β-cell differentiation and regeneration; it suppresses glucagon release; it suppresses gastric emptying and the urge to eat; and it improves sensitivity to insulin receptors. GLP-1 has been clinically proven to have benefits such as reducing the risk of hypoglycemia, promoting weight loss, and enhancing beneficial effects on the cardiovascular system.

[0005] The combined use of insulin and GLP-1 receptor agonists allows for the full utilization of their complementary clinical benefits, such as blood glucose reduction, weight loss, reduced risk of hypoglycemia, and reduced insulin resistance. Premix formulations containing basal insulin and GLP-1 are easy to administer and therefore have high market demand.

[0006] However, there is still room for improvement in the current pharmaceutical compositions of GLP-1 receptor agonists. [Overview of the Initiative]

[0007] One aspect of the present invention relates to a stable pharmaceutical composition comprising a first active ingredient containing one or more GLP-1 receptor agonists and one or more surfactants (for example, but not limited to alkyl glycosides (for example, but not limited to n-dodecyl β-D-maltoside (DDM))).

[0008] Embodiments of stable pharmaceutical compositions disclosed herein demonstrate improved stability of one or more GLP-1 receptor agonists. In some embodiments, the improved stability relates to physical stability. In some embodiments, the stable pharmaceutical composition is a stable liquid pharmaceutical composition. In some embodiments, the stable liquid pharmaceutical composition is a stable aqueous pharmaceutical composition. In some embodiments, the stable aqueous pharmaceutical composition is a stable aqueous pharmaceutical composition for injection. In some embodiments, the stable aqueous injectable pharmaceutical composition is an aqueous injectable pharmaceutical composition for single-dose or multiple-dose administration. Examples of the one or more GLP-1 receptor agonists include, but are not limited to, GLP-1, GLP-1 analogs, and derivatives of GLP-1 analogs.

[0009] In some embodiments, the stable pharmaceutical composition further comprises insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient, and may be referred to herein as a stable combination pharmaceutical composition. In the embodiments of the stable combination pharmaceutical compositions disclosed herein, both the first and second active ingredients exhibit high stability. In some embodiments, the stability refers to physical stability. In some embodiments, the stable combination pharmaceutical composition is a stable liquid combination pharmaceutical composition. In some embodiments, the stable liquid-compound pharmaceutical composition is a stable aqueous-compound pharmaceutical composition. In some embodiments, the stable aqueous combination pharmaceutical composition is a stable aqueous combination pharmaceutical composition for injection. In some embodiments, the stable aqueous injectable combination pharmaceutical composition is an aqueous injectable combination pharmaceutical composition for single-dose or multiple-dose administration.

[0010] Another aspect of the present invention relates to a method for improving the storage stability of a pharmaceutical composition containing a first active ingredient comprising one or more GLP-1 receptor agonists. In some embodiments of the methods disclosed herein, the method includes the step of mixing one or more surfactants (e.g., alkyl glycosides (e.g., DDM, but not limited to)) with the pharmaceutical composition. In some embodiments, the pharmaceutical composition is a combination pharmaceutical composition further comprising insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient.

[0011] Another aspect of the present invention relates to the use of a stable pharmaceutical composition in the manufacture of pharmaceuticals used in the following applications: (i) prevention and / or treatment of diabetes mellitus and / or reduction of HbA1c; (ii) delay or prevention of the progression of diabetes mellitus, delay of progression from impaired glucose tolerance to type 2 diabetes mellitus requiring insulin, delay or prevention of insulin resistance and / or delay of progression from type 2 diabetes mellitus that does not require insulin to type 2 diabetes mellitus that requires insulin; (iii) improvement of β-cell function and / or restoration of glucose sensitivity of β-cells; (iv) prevention and / or treatment of eating disorders, decreased gastric motility, and delayed gastric emptying; (v) prevention and / or treatment of complications associated with obesity; (vi) prevention and / or treatment of complications associated with diabetes mellitus; and / or (vii) prevention and / or treatment of cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient.

[0012] Another aspect of the present invention relates to the use of a stable pharmaceutical composition for the following purposes: (i) prevention and / or treatment of diabetes mellitus, and / or reduction of HbA1c; (ii) delay or prevention of the progression of diabetes mellitus, delay of progression from impaired glucose tolerance to type 2 diabetes mellitus requiring insulin, delay or prevention of insulin resistance, and / or delay of progression from type 2 diabetes mellitus that does not require insulin to type 2 diabetes mellitus that requires insulin; (iii) improvement of β-cell function and / or restoration of glucose sensitivity of β-cells; (iv) prevention and / or treatment of eating disorders, decreased gastric motility, and delayed gastric emptying; (v) prevention and / or treatment of complications associated with obesity; (vi) prevention and / or treatment of complications associated with diabetes mellitus; and / or (vii) prevention and / or treatment of cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient.

[0013] Another aspect of the present invention relates to a method used for the following purposes, which includes administering a therapeutically effective amount of a stable pharmaceutical composition to a subject: (i) prevention and / or treatment of diabetes and / or reduction of HbA1c; (ii) delay or prevention of the progression of diabetes, delay of the progression from impaired glucose tolerance to type 2 diabetes requiring insulin administration, delay or prevention of insulin resistance, and / or delay of the progression from type 2 diabetes not requiring insulin to type 2 diabetes requiring insulin administration; (iii) improvement of β-cell function and / or restoration of β-cell glucose sensitivity; (iv) prevention and / or treatment of eating disorders, reduction of gastric motility, delay of gastric emptying; (v) prevention and / or treatment of complications associated with obesity; (vi) prevention and / or treatment of complications associated with diabetes; and / or (vii) prevention and / or treatment of cardiovascular diseases. In some embodiments, the stable pharmaceutical composition is a stable combined pharmaceutical composition further comprising insulin, an insulin analog and / or a derivative of an insulin analog as a second active ingredient.

[0014] The above-mentioned and / or further other aspects and advantages of the present disclosure will become apparent and be easily understood by those skilled in the art upon reading the description of the embodiments while referring to the following drawings.

Brief Description of the Drawings

[0015] [Figure 1] Figure 1 shows the ThT test results of an insulin glargine / GLP-1 combined composition according to an embodiment of the present disclosure. [Figure 2] Figure 2 shows the ThT test results of a venglutide stock solution according to an embodiment of the present disclosure. [Figure 3] Figure 3 shows the ThT test results of venglutide compositions without DDM and containing different concentrations of DDM according to an embodiment of the present disclosure. [Figure 4] Figure 4 shows the ThT test results of an insulin glargine / GLP-1-DDM combined composition containing different preservatives according to an embodiment of the present disclosure. [Figure 5]Figure 5 shows the ThT test results of an insulin glargine / GLP-1 combination composition according to an embodiment of the present disclosure. [Figure 6] Figure 6 (SEQ ID NO: 5) shows the physical stability of GLP-1 pharmaceutical compositions with and without 0.1% DDM according to multiple embodiments of the present disclosure. [Figure 7] Figure 7 shows the physical stability of a P13 (peptide sequence: ELAEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2 (SEQ ID NO: 3), GLP-1 / GIP dual agonist) pharmaceutical composition with and without 0.1% DDM according to multiple embodiments of the present disclosure. [Figure 8] Figure 8 shows the physical stability of 988927 (peptide sequence: HGEGTFTSDSSSYLEEQAAKEFIAWLVKGRG (SEQ ID NO: 4)) with and without 0.1% DDM according to multiple embodiments of the present disclosure. [Figure 9] Figure 9 shows the solubility of GLP-1 with respect to phenol concentration with and without DDM. [Figure 10] Figure 10 shows the plasma concentration of insulin aspart in beagle dogs after administration of an insulin aspart monocomposition and an insulin aspart / GLP-1 combination composition according to an embodiment of the present disclosure. [Figure 11] Figure 11 shows the plasma concentration of GLP-1 in beagle dogs after administration of a GLP-1 monocomposition and an insulin aspart / GLP-1 combination composition according to an embodiment of the present disclosure.

Embodiments for Carrying Out the Invention

[0016] Hereinafter, embodiments of the present disclosure will be described in detail. The embodiments described below with reference to the drawings are examples for the purpose of explaining the present disclosure and should not be construed as limiting the present disclosure.

[0017] Definitions Unless otherwise specified, the term “active ingredient” as used herein refers to a substance or combination of substances in a pharmaceutical product that exerts a therapeutic or preventive effect. For example, according to embodiments of this application, the active ingredient may be one or more GLP-1 receptor agonists, and insulin or its analogs or derivatives may be added to the pharmaceutical composition. Such a configuration allows the pharmaceutical composition to exert the desired therapeutic or preventive effect when applied to a subject.

[0018] Unless otherwise specified, the term “surfactant” as used herein refers to a group of chemical substances that can reduce the surface tension and interfacial tension of a liquid and are typically composed of hydrophobic and hydrophilic groups. Surfactants are typically used to modulate the solubility, stability, and bioavailability of drugs, thereby influencing their absorption, distribution, metabolism, and excretion.

[0019] Unless otherwise specified, the term “alkyl glycoside” as used herein refers to a type of surfactant composed of a sugar group and an alkyl chain. These possess good surface activity and biodegradability. These compounds are typically obtained by enzymatically catalyzing the reaction of a sugar with an aliphatic alcohol under a given temperature and pressure. Because the molecular structure of alkyl glycosides contains both hydrophobic and hydrophilic groups, they have properties similar to conventional surfactants. In this context, the alkyl chain is hydrophobic and can interact with nonpolar solvents, while the sugar group is hydrophilic and can interact with water. The inventors of this disclosure have found that by using alkyl glycosides (e.g., DDM, but not limited to DDM) as nonionic surfactants when preparing the pharmaceutical compositions of this application, the stability, solubility, and bioavailability of the drug can be improved, and the toxicity and irritancy of the formulation can be reduced, thereby improving the safety and tolerability of the drug.

[0020] Unless otherwise specified, the term “analog” as used herein refers to a molecule that is structurally similar to the original molecule but has the same or different chemical properties. For example, in pharmaceutical research, researchers may design and synthesize a series of compounds that are structurally similar to known drug molecules but have different chemical groups or substituents, thereby altering their pharmacological properties. These properties may include, for example, absorption, metabolism, and efficacy. These analogs may have superior pharmacological or toxicological properties and therefore have the potential to be further developed as pharmaceuticals. More specifically, in this application, “analog” means a peptide in which one or more amino acid residues of the parent peptide are substituted with other amino acid residues, and / or a peptide in which one or more amino acid residues of the parent peptide are deleted, and / or a peptide in which one or more amino acid residues are added to the parent peptide. Such additions usually occur at the N-terminus, C-terminus, or both of the parent peptide. In particular embodiments, the term "analog" in this application generally refers to a peptide in which six or fewer amino acids of the parent peptide are substituted and / or added and / or deleted, more preferably a peptide in which three or fewer amino acids of the parent peptide are substituted and / or added and / or deleted, and most preferably a peptide in which one amino acid of the parent peptide is substituted and / or added and / or deleted.

[0021] Unless otherwise specified, the term “derivative” as used herein refers to a new molecule obtained from an original molecule through chemical modification or transformation. For example, in pharmaceutical research, researchers can synthesize a series of novel compounds by changing, substituting, or modifying the structure of a known pharmaceutical molecule. These novel molecules are referred to as derivatives of the pharmaceutical molecule. Derivatives play an important role in drug development because they may have superior pharmacological properties, higher safety, or lower side effects. Specifically, in this application, “derivative” refers to a peptide in which one or more amino acid residues of the parent peptide have been substituted. Typical substituents include amides, sugars, alkyls, acyls, esters, and PEGylated groups.

[0022] Unless otherwise specified, the term "GLP-1 receptor agonist" as used herein refers to a group of drugs that treat diabetes by mimicking the biological activity of glucagon-like polypeptide 1 (GLP-1), thereby promoting insulin secretion and suppressing glucose production and release, thus lowering blood glucose levels. GLP-1 is a peptide hormone secreted from intestinal L cells and brainstem neurons. GLP-1 has diverse biological functions, including appetite suppression, stimulation of insulin secretion, and suppression of glucose production and release. GLP-1 receptor agonists can enhance insulin secretion and suppress glucose production and release by stimulating the GLP-1 receptor. Due to their efficacy in blood glucose control and good tolerability, GLP-1 receptor agonists have become one of the first-line treatments for type 2 diabetes. In addition to diabetes treatment, GLP-1 receptor agonists are also being studied in other fields such as obesity, cardiovascular disease, and neurodegenerative diseases.

[0023] Unless otherwise specified, “treatment” as used herein means the use of drugs to alleviate or cure a disease condition, and “prevention” means the use of drugs to prevent the onset of a disease or reduce the likelihood of the disease occurring, and includes preventing or delaying the onset or progression of a disease.

[0024] In general, alkyl glycosides (e.g., DDMs, but not limited to them) have been used as transmucosal absorption enhancers for polypeptides, proteins, and low molecular weight compounds. However, in various embodiments of the stable pharmaceutical compositions disclosed herein, the presence of alkyl glycosides (e.g., DDMs, but not limited to them) was not expected to improve the stability of one or more active ingredients (e.g., GLP-1 receptor agonists, and insulin, insulin analogs, and / or derivatives of insulin analogs). In some embodiments of the pharmaceutical compositions disclosed herein, the stability of one or more active ingredients is sufficiently high, and as a result, these embodiments can be used as aqueous injectable compositions for single or multiple doses.

[0025] Pharmaceutical compositions of GLP-1 receptor agonists, particularly liquid compositions thereof, tend to form fibrous substances during storage, making it difficult to obtain pharmaceutical compositions with long-term stability. As shown in the examples herein, it was not anticipated that certain surfactants would improve the long-term stability of GLP-1 receptor agonists in certain embodiments of the pharmaceutical compositions disclosed herein. In particular, surfactants such as alkyl glycosides (e.g., DDM, but not limited to these) improved the stability of GLP-1 receptor agonists in various embodiments of aqueous pharmaceutical compositions.

[0026] One aspect of the present invention relates to a stable pharmaceutical composition comprising a first active ingredient containing one or more GLP-1 receptor agonists and one or more surfactants.

[0027] In some embodiments, the surfactant comprises an alkyl glycoside (e.g., DDM, but not limited to these). Embodiments of stable pharmaceutical compositions disclosed herein demonstrate improved stability of one or more GLP-1 receptor agonists. In some embodiments of the stable pharmaceutical compositions disclosed herein, when the GLP-1 receptor agonist comprises, for example, GLP-1 or its analog or derivative, the presence of an alkyl glycoside (e.g., DDM, but not limited to) can suppress the formation of GLP-1 fibrillation, thereby improving the physical stability of the GLP-1 receptor agonist. Alkyl glycosides (e.g., DDM, but not limited to) are significantly superior to other types of surfactants.

[0028] In some embodiments, the improved stability relates to physical stability. In some embodiments, the stable pharmaceutical composition is a stable liquid pharmaceutical composition. In some embodiments, the stable liquid pharmaceutical composition is a stable aqueous pharmaceutical composition. In some embodiments, the stable aqueous pharmaceutical composition is a stable aqueous pharmaceutical composition for injection. In some embodiments, the stable aqueous injectable pharmaceutical composition is an aqueous injectable pharmaceutical composition for single-dose or multiple-dose administration.

[0029] Examples of the one or more GLP-1 receptor agonists mentioned above include, but are not limited to, GLP-1, GLP-1 analogs, and derivatives of GLP-1 analogs.

[0030] In some embodiments, the stable pharmaceutical composition further comprises insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient, and may be referred to herein as a stable combination pharmaceutical composition. In the embodiments of the stable combination pharmaceutical compositions disclosed herein, both the first and second active ingredients exhibit high stability. In some embodiments, the stability refers to physical stability. In some embodiments, the stable combination pharmaceutical composition is a stable liquid combination pharmaceutical composition. In some embodiments, the stable liquid-compound pharmaceutical composition is a stable aqueous-compound pharmaceutical composition. In some embodiments, the stable aqueous combination pharmaceutical composition is a stable aqueous combination pharmaceutical composition for injection. In some embodiments, the stable aqueous injectable combination pharmaceutical composition is an aqueous injectable combination pharmaceutical composition for single-dose or multiple-dose administration.

[0031] In some embodiments of the stable combination pharmaceutical compositions disclosed herein, the second active ingredient comprises insulin or an insulin analog. In some embodiments of the stable combination pharmaceutical compositions disclosed herein, the insulin analog comprises at least one selected from insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin degludec, and insulin detemir. In some embodiments of the stable combination pharmaceutical compositions disclosed herein, the stable combination pharmaceutical composition comprises about 0.1% to about 1% by mass of insulin, an insulin analog, or a derivative of an insulin analog, based on the total weight of the stable combination pharmaceutical composition, and optionally comprises at least about 0.1% to about 0.4% by mass of insulin, an insulin analog, or a derivative of an insulin analog, for example, at least about 0.2% to about 0.4% by mass of insulin or an insulin analog. For example, the stable combination pharmaceutical composition comprises at least one of insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin degludec, and insulin detemir, and its concentration may be at least about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, or about 1.0%. In some embodiments, the stable combination pharmaceutical composition includes insulin, insulin analogs, or derivatives of insulin analogs having a concentration of at least about 0.20% by mass to about 0.40% by mass, for example, but not limited to, about 0.21% by mass, about 0.22% by mass, about 0.23% by mass, about 0.24% by mass, about 0.25% by mass, about 0.26% by mass, about 0.27% by mass, about 0.28% by mass, about 0.29% by mass, about 0.30% by mass, about 0.31% by mass, about 0.32% by mass, about 0.33% by mass, about 0.34% by mass, about 0.35% by mass, about 0.36% by mass, about 0.37% by mass, about 0.38% by mass, about 0.39% by mass, or about 0.40% by mass. Here, it should be noted that when the stable combination pharmaceutical composition is configured as a liquid pharmaceutical composition such as a premix aqueous injection composition, the above-mentioned "mass%" can also be expressed in "mg / mL". For example, 0.40 mass% of insulin or insulin analog or insulin analog derivative is equivalent to 4.0 mg / mL of insulin or insulin analog or insulin analog derivative. As shown in the examples described in the Examples section, alkyl glycosides (e.g., DDM, but not limited to them) unexpectedly improved the stability of GLP-1 receptor agonists, and simultaneously improved the stability of both GLP-1 receptor agonists and insulin or insulin analogs or derivatives of insulin analogs.

[0032] In some embodiments of the stable pharmaceutical compositions disclosed herein, an example of an alkyl glycoside comprises a sugar group and an alkyl group linked by a linking group, wherein the sugar group comprises at least one sugar selected from glucose, maltose, sucrose, or trehalose, the alkyl group comprises about 10 to about 16 carbon atoms, and the linking group comprises at least one bond selected from a glycosidic bond, a thioglycosidic bond, or an amide bond. In some embodiments, the alkyl group contains about 10 to about 14 carbon atoms. In some embodiments, the alkyl group contains about 6 to about 20 carbon atoms. In some embodiments, the alkyl group contains 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. In some embodiments, the alkyl group includes linear alkyl and / or branched alkyl groups. In some embodiments, the alkyl group includes a lower alkyl group (e.g., C1-C8) substituted with a cycloalkyl group. Examples of the lower alkyl group include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, hexyl, isohexyl, heptyl, isoheptyl, octyl, and isooctyl. Examples of cycloalkyl groups include, but are not limited to, cyclohexyl. In some embodiments, alkyl glycosides (e.g., DDM, but not limited to) improve the stability of the stable pharmaceutical compositions disclosed herein. In some embodiments, the sugar group comprises maltose or sucrose. In some embodiments, the alkyl glycoside is n-dodecyl-β-D-maltoside (DDM), sucrose monododecanoate, n-dodecyl-β-D-maltoside, n-hexyl-β-D-glucoside, n-heptyl-β-D-glucoside, n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-glucoside, 3-cyclohexyl-1-propyl-β-D-glucoside, n-hexyl-β-D - comprises at least one compound selected from glucopyranoside, n-octyl-β-D-maltoside, n-nonyl-β-D-maltoside, n-decyl-β-D-maltoside, cyclohexyl-methyl-β-D-maltoside, 2-cyclohexyl-ethyl-β-D-maltoside, 3-cyclohexyl-propyl-β-D-maltoside, 4-cyclohexyl-butyl-β-D-maltoside, or 5-cyclohexyl-amyl-β-D-maltoside.

[0033] In some embodiments, based on the total weight of the stable pharmaceutical composition, the stable pharmaceutical composition contains about 0.05% to about 0.50% by mass of alkyl glycoside. For example, the concentration of the alkyl glycoside is at least about 0.05% by mass, about 0.06% by mass, about 0.07% by mass, about 0.08% by mass, about 0.09% by mass, about 0.10% by mass, about 0.11% by mass, about 0.12% by mass, about 0.13% by mass, about 0.14% by mass, about 0.15% by mass, about 0.16% by mass, about 0.17% by mass, about 0.18% by mass, about 0.19% by mass, about 0.20% by mass, about 0.21% by mass, about 0.22% by mass, about 0.23% by mass, about 0.24% by mass, about 0.25% by mass, about 0.26% by mass, about 0.27 mass%, approximately 0.28 mass%, approximately 0.29 mass%, approximately 0.30 mass%, approximately 0.31 mass%, approximately 0.32 mass%, approximately 0.33 mass%, approximately 0.34 mass%, approximately 0.35 mass%, approximately 0.36 mass%, approximately 0.37 mass%, approximately 0.38 mass%, approximately 0.39 % by weight, about 0.40% by weight, about 0.41% by weight, about 0.42% by weight, about 0.43% by weight, about 0.44% by weight, about 0.45% by weight, about 0.46% by weight, about 0.47% by weight, about 0.48% by weight, about 0.49% by weight, or about 0.50% by weight. In some embodiments, the concentration of the alkyl glycoside is at least about 0.10% by mass to about 0.20% by mass. In some embodiments, the concentration of the alkyl glycoside is at least about 0.10% by mass to about 0.30% by mass. In some embodiments of the stable combination pharmaceutical compositions disclosed herein, the stable pharmaceutical composition is a stable combination pharmaceutical composition comprising one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs, and derivatives of GLP-1 analogs such as bainaglutide), a second active ingredient comprising or consisting of insulin aspart, a surfactant comprising one or more alkyl glycosides (e.g., DDM, but not limited to), and the concentration of the surfactant being about 0.05% to about 0.50% by mass, about 0.05% to about 0.45% by mass, about 0.05% to about 0.40% by mass, about 0.05% to about 0.35% by mass, about 0.05% to about 0.30% by mass, and about 0.05% by mass. Mass% to approximately 0.25 mass%, 0.05 mass% to approximately 0.20 mass%, approximately 0.05 mass% to approximately 0.15 mass%, 0.05 mass% to approximately 0.10 mass%, 0.10 mass% to approximately 0.50 mass%, approximately 0.10 mass% to approximately 0.45 mass%, approximately 0.10 mass% to approximately 0.40 mass%, approximately 0.10 mass% to approximately 0.35 mass%, 0.10 mass% to approximately 0.30 mass%, approximately 0.10 mass% to approximately 0.2 5 mass%, 0.10 mass% to approximately 0.20 mass%, approximately 0.10 mass% to approximately 0.15 mass%, approximately 0.16 mass% to approximately 0.30 mass%, approximately 0.16 mass% to approximately 0.25 mass%, approximately 0.20 mass% % to about 0.30% by weight, about 0.20% to about 0.25% by weight, about 0.05% by weight, about 0.10% by weight, about 0.16% by weight, about 0.20% by weight or about 0.25% by weight.

[0034] In some embodiments, the GLP-1 receptor agonist is a polypeptide agonist. Optionally, the GLP-1 receptor agonist may also function as a GIP agonist. In other words, a dual agonist (e.g., P13, but not limited to P13) may be used.

[0035] In some embodiments, the GLP-1 receptor agonist comprises at least one selected from the following: GLP-1, a GLP-1 analog or its derivative, and a cleaved GLP-1 fragment or its analog or derivative. Examples of GLP-1 analogs include, but are not limited to, 988927, exendin 4 and its analogs and derivatives. In some embodiments, the GLP-1 receptor agonist comprises at least one selected from liraglutide, exenatide, lixisenatide, albiglutide, vainaglutide, doxepinlaglutide, semaglutide, pepamotidide, and tilpotide. In some embodiments, the GLP-1 receptor agonist comprises or consists of vainaglutide.

[0036] As used herein, “GLP-1 receptor agonists” refers to polypeptides comprising native, extended, or cleaved GLP-1 polypeptides (e.g., GLP-1(7-37)OH / NH2, GLP-1(7-36)OH / NH2, GLP-1(7-35)OH / NH2), GLP-1 fragments, GLP-1 analogs, and derivatives thereof. GLP-1 receptor agonists may also include dual or multiple agonists based on the GLP-1 sequence, such as, but not limited to, GLP-1 / GIP dual agonists (e.g., P13). The GLP-1 compounds or GLP-1 receptor agonists used herein can bind to the GLP-1 receptor and thereby initiate a signaling pathway that results in insulin secretion-promoting activity. GLP-1 and GLP-1 analogs may have a free carboxyl group at the C-terminus or an amide group at the C-terminus. For example, a GLP-1 analog may be a recombinant human GLP-1(7-36) peptide having the following sequence: His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg (SEQ ID NO: 5), which is referred to as bainaglutide. The molecular formula for baynaglutide is C 149 H 225 N 39 O 46 Its molecular weight is 3,298.7. Beinaglutide is essentially identical to the active form of GLP-1 in vivo, but differs in that, except for the endogenous amidation, the NH2 group present at the C-terminus in the native form is replaced with an OH group in the recombinant peptide. Beinaglutide has a free carboxyl group at its C-terminus.

[0037] In some embodiments of the stable pharmaceutical compositions disclosed herein, the GLP-1 receptor agonist is a polypeptide having the following sequence:XHXEGTXTSDXSXXXEXXAXXXFIXWLXXGXX(SEQ ID NO: 1). Here, X at position 1 is either R or missing. X in position 3 is A, G, V, L, I, S, or T. X at position 7 is F, W, or Y. X at position 11 is V, S, W, I, L, K, F, or Y. The X in position 13 is S, W, Y, F, K, I, L, or V. The X at position 14 is either Y, W, or F. The X of position 15 is L, F, Y, or W. The X at position 17 is G, E, D, Q, N, K, R, or C. X at position 18 is H, D, K, E, or Q. X at position 20 is A, V, I, or L. X at position 21 is K, R, Q, or N. The X of position 22 is A, E, H, F, Y, W, R, I, or K. X at position 25 is A, E, D, S, or H. At position 28, X is either V or I. X at position 29 is K, R, Q, or N. X at position 31 is R, R-NH2, K, or K-NH2. X at position 32 is G, K, R, T, S, E, D, W, Y, F, H, or missing.

[0038] In some embodiments of the stable pharmaceutical compositions disclosed herein, the GLP-1 receptor agonist is a polypeptide having the following sequence:XXXGTXXXXXSKQXEEEAVXLXXXXLKNGGXXXXXXXXXX(SEQ ID NO: 2). Here, The X coordinate at position 1 is H, R, Y, or EL. X at position 2 is S, G, A, or T. X at position 3 is either D or E. At position 6, X is either F or Y. At position 7, X is T, Y, or S. At position 8, X is either S or Y. X at position 9 is either D or E. At position 10, X is either L or I. X at position 14 is L, I, V, or M. At position 20, X is either R or K. The X value at position 22 is either F or Y. X at position 23 is I, V, L, or M. At position 24, X is either E or D. At position 25, X is W, F, or Y. X at position 31 is either P or missing. X at position 32 is either S or a deletion. X at position 33 is either S or a deletion. X at position 34 is either G or missing. X at position 35 is either A or missing. X at position 36 is either P or missing. X at position 37 is either P or missing. X at position 38 is either P or missing. X at position 39 is S, R, or deletion. X at position 40 is any amino acid or a deletion.

[0039] In general, the stability of GLP-1 receptor agonists can be impaired by various stresses. These stresses include pH, temperature, light, shaking, and gas-liquid interfaces. Various common surfactants were also tested to investigate the possibility of improving the stability of pharmaceutical compositions containing a first active ingredient (e.g., a GLP-1 receptor agonist) and / or a second active ingredient (e.g., insulin, insulin analogs, and derivatives of insulin analogs). Examples of surfactants that were tested but were not effective in improving stability include, for example, polysorbate 20, polysorbate 80, poloxamer 188, hydroxypropyl β-cyclodextrin, and amino acids. Furthermore, embodiments of stable pharmaceutical compositions that do not contain any surfactant selected from the group consisting of polysorbate 20, polysorbate 80, poloxamer 188, and hydroxypropyl β-cyclodextrin are also provided herein. In some embodiments, the stable pharmaceutical composition does not contain polysorbate 20, polysorbate 80, poloxamer 188, hydroxypropyl β-cyclodextrin, or amino acids. In some embodiments, the stable pharmaceutical composition does not contain surfactants other than one or more alkyl glycosides (e.g., DDM, but not limited to these).

[0040] In some embodiments, the stable pharmaceutical composition is a liquid. In some embodiments, the stable pharmaceutical composition is a premixed aqueous injection. In some embodiments, the stable pharmaceutical composition is configured as a premix formulation which is an injectable composition, and is preferably for subcutaneous administration.

[0041] In some embodiments of the stable pharmaceutical compositions disclosed herein, the pharmaceutical compositions optionally include other pharmaceutically acceptable excipients, such as, but not limited to, buffers, preservatives, pharmaceutically acceptable solubilizers, and zinc ions.

[0042] In some embodiments, the pH of the stable pharmaceutical compositions disclosed herein is about 3.0 to about 9.0, about 3.0 to about 5.0, about 4.0 to about 5.0, or about 4.0 to about 4.5. In some embodiments, the pH of the stable pharmaceutical compositions disclosed herein is approximately 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, and 5. 0.7, approximately 5.8, approximately 5.9, approximately 6.0, approximately 6.1, approximately 6.2, approximately 6.3, approximately 6.4, approximately 6.5, approximately 6.6, approximately 6.7, approximately 6.8, approximately 6.9, approximately 7.0, approximately 7.1, approximately 7.2, approximately 7.3, approximately 7.4, approximately 7.5, approximately 7.6, approximately 7.7, approximately 7.8, approximately 7.9, approximately 8.0, approximately 8.1, approximately 8.2, approximately 8.3, approximately 8.4, approximately 8.5, approximately 8.6, approximately 8.7, approximately 8.8, approximately 8.9, or approximately 9.0.

[0043] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs and derivatives of GLP-1 analogs such as bainaglutide), or its pH is about 3.0 to about 4.5, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, or about 4.5.

[0044] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs, and derivatives of GLP-1 analogs, such as bainaglutide), and its pH is approximately 6.5 to approximately 8.5, approximately 6.6 to approximately 8.4, approximately 6.7 to approximately 8.3, approximately 6.8 to approximately 8.2, approximately 6.9 to approximately 8.2, approximately 7.0 to approximately 8.2, approximately 7.0 to approximately 8.15, approximately 7.0 to approximately 8.1, approximately 7.0 to approximately 8.0, approximately 7.0 to approximately 7.9, approximately 7.0 to approximately 7.8, approximately 7.0 to approximately 7.7, and approximately 7. 0 to approximately 7.6, approximately 7.1 to approximately 8.2, approximately 7.1 to approximately 8.15, approximately 7.1 to approximately 8.1, approximately 7.1 to approximately 8.0, approximately 7.1 to approximately 7.9, approximately 7.1 to approximately 7.8, approximately 7.1 to approximately 7.7, approximately 7.1 to approximately 7.6, approximately 7.2 to approximately 8.2, approximately 7.2 to approximately 8.15, approximately 7.2 to approximately 8.1, approximately 7.2 to approximately 8. 0, approximately 7.2-7.9, approximately 7.2-7.8, approximately 7.2-7.7, approximately 7.2-7.6, approximately 7.3-8.2, approximately 7.3-8.15, approximately 7.3-8.1, approximately 7.3-8.0, approximately 7.3-7.9, approximately 7.3-7.8, approximately 7.3-7.7, or approximately 7.3-7.6.

[0045] In some embodiments, the stable pharmaceutical composition is the stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs and derivatives of GLP-1 analogs such as bainaglutide), and the second active ingredient comprises or consists of insulin glargine, with a pH of approximately 3.5 to approximately 4.6, approximately 3.5 to approximately 4.5, approximately 3.5 to approximately 4.4, approximately 3.5 to approximately 4.3, approximately 3.5 to approximately 4.2, approximately 3.5 to approximately 4.1, approximately 3.5 to approximately 4.0, 3.6 to approximately 4.6, approximately 3.6 to approximately 4.5, and approximately 3. 6~approx. 4.4, approx. 3.6~approx. 4.3, approx. 3.6~approx. 4.2, approx. 3.6~approx. 4.1, approx. 3.6~approx. 4.0, 3.7~approx. 4.6, approx. 3.7~approx. 4.5, approx. 3.7~approx. 4.4, approx. 3.7~approx. 4.3, approx. 3.7~approx. 4.2, approx. 3.7~approx. 4.1, approx. 3.7~approx. 4.0, approx. 3.8~approx. 4.6, approx. 3.8 The ranges are approximately 4.5, 3.8 to 4.4, 3.8 to 4.3, 3.8 to 4.2, 3.8 to 4.1, 3.8 to 4.0, 3.9 to 4.6, 3.9 to 4.5, 3.9 to 4.4, 3.9 to 4.3, 3.9 to 4.2, 3.9 to 4.1, or 3.9 to 4.0.

[0046] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs and derivatives of GLP-1 analogs such as bainaglutide), and the second active ingredient comprises or consists of insulin aspart, with a pH of approximately 6.5 to approximately 8.5, approximately 6.6 to approximately 8.4, approximately 6.7 to approximately 8.3, approximately 6.8 to approximately 8.2, approximately 6.9 to approximately 8.2, approximately 7.0 to approximately 8.2, approximately 7.0 to approximately 8.15, approximately 7.0 to approximately 8.1, approximately 7.0 to approximately 8.0, approximately 7.0 to approximately 7.9, and approximately 7.0 ~7.8, 7.0~7.7, 7.0~7.6, 7.1~8.2, 7.1~8.15, 7.1~8.1, 7.1~8.0, 7.1~7.9, 7.1~7.8, 7.1~7.7, 7.1~7.6, 7.2~8.2, 7.2~8.15, 7.2~8.1 These are approximately 7.2-8.0, 7.2-7.9, 7.2-7.8, 7.2-7.7, 7.2-7.6, 7.3-8.2, 7.3-8.15, 7.3-8.1, 7.3-8.0, 7.3-7.9, 7.3-7.8, 7.3-7.7, or 7.3-7.6.

[0047] In some embodiments, the stable pharmaceutical composition includes one or more buffer systems commonly used for pH adjustment. The buffer system may include one or more buffer salts. Commonly used buffer salts include, but are not limited to, acetates, chlorides, TRIS, HEPES, MOPS, PIPES, BES, Bis-Tris, TES, and / or phosphates. The concentration of the buffer salt (for example, but not limited to acetates, chlorides, and phosphates) may be about 0 mM to about 50 mM, about 0 mM to about 45 mM, about 0 mM to about 40 mM, about 0 mM to about 35 mM, about 0 mM to about 30 mM, about 0 mM to about 25 mM, about 0 mM to about 20 mM, about 10 mM to about 50 mM, about 10 mM to about 45 mM, about 10 mM to about 40 mM, about 10 mM to about 35 mM, about 10 mM to about 30 mM, about 10 mM to about 25 mM, or about 10 mM to about 20 mM. In some embodiments, the stable pharmaceutical composition may not contain a buffer salt.

[0048] In some embodiments, the stable pharmaceutical composition contains zinc ions. In some embodiments, the zinc ion concentration range is approximately 1 μg / mL to approximately 100 μg / mL, approximately 10 μg / mL to approximately 70 μg / mL, approximately 10 μg / mL to approximately 50 μg / mL, approximately 13 μg / mL to approximately 33 μg / mL, approximately 20 μg / mL to approximately 40 μg / mL, approximately 25 μg / mL to approximately 40 μg / mL, approximately 10 μg / mL, approximately 15 μg / mL, approximately 20 μg / mL, approximately 25 μg / mL, approximately 30 μg / mL, approximately 35 μg / mL, approximately 40 μg / mL, approximately 45 μg / mL, or approximately 50 μg / mL. In some embodiments, the molar ratio of zinc ions to a second active ingredient (e.g., insulin, an insulin analog, or a derivative of an insulin analog) is approximately 1:4 to approximately 10.8:6.

[0049] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs and derivatives of GLP-1 analogs such as bainaglutide), and the second active ingredient comprises insulin glargine or comprises insulin glargine, and further, the stable combination pharmaceutical composition has concentrations of approximately 1 μg / mL to approximately 100 μg / mL, approximately 10 μg / mL to approximately 70 μg / mL, approximately 10 μg / mL to approximately 50 μg / mL, approximately 13 μg / mL to approximately 33 μg / mL, approximately 20 μg / mL to approximately 40 μg / mL, approximately 25 μg / mL to approximately 40 μg / mL, approximately 10 μg / mL, approximately 15 μg / mL, approximately 20 μg / mL, approximately 25 μg / mL, approximately 30 μg / mL, approximately 35 μg / mL, and approximately 40 μg / mL. It further contains zinc ions in the form of μg / mL, approximately 45 μg / mL, or approximately 50 μg / mL. In some embodiments, the molar ratio of zinc ions to a second active ingredient (e.g., insulin, an insulin analog, or a derivative of an insulin analog) is approximately 1:4 to approximately 10.8:6.

[0050] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs and derivatives of GLP-1 analogs such as bainaglutide), and the second active ingredient comprises or consists of insulin aspart, and further, the stable combination pharmaceutical composition has concentrations of approximately 1 μg / mL to approximately 100 μg / mL, approximately 10 μg / mL to approximately 70 μg / mL, approximately 10 μg / mL to approximately 50 μg / mL, approximately 13 μg / mL to approximately 33 μg / mL, approximately 20 μg / mL to approximately 40 μg / mL, approximately 25 μg / mL to approximately 40 μg / mL, approximately 10 μg / mL, approximately 15 μg / mL, approximately 20 μg / mL, approximately 25 μg / mL, approximately 30 μg / mL, approximately 35 μg / mL, and approximately 40 It further contains zinc ions in the form of μg / mL, approximately 45 μg / mL, or approximately 50 μg / mL. In some embodiments, the molar ratio of zinc ions to a second active ingredient (e.g., insulin, an insulin analog, or a derivative of an insulin analog) is approximately 1:4 to approximately 10.8:6.

[0051] In some embodiments, the stable pharmaceutical composition may contain one or more anions. Examples of such anions include, but are not limited to, chloride ions, thiocyanate ions (KSCN), para-aminobenzoate ions (PABA), and 1-hydroxy-3-nitrobenzoate ions (4H3N). In some embodiments, the concentration of the anion (e.g., chloride ions or other examples disclosed herein) is at least about 10 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, at least about 50 mM, at least about 60 mM, at least about 70 mM, at least about 80 mM, at least about 90 mM, or at least about 100 mM.

[0052] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition disclosed herein, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, GLP-1 analogs and derivatives of GLP-1 analogs such as bainaglutide), and the second active ingredient comprises or comprises insulin aspart or insulin glargine, and further, the stable combination pharmaceutical composition comprises an anion, for example, Cl - KSCN - PABA - and 4H3N - Includes. In some embodiments, the concentration of the anion (e.g., chloride ions or other examples disclosed herein) is at least about 10 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, at least about 50 mM, at least about 60 mM, at least about 70 mM, at least about 80 mM, at least about 90 mM, or at least about 100 mM.

[0053] In some embodiments, the stable pharmaceutical composition comprises one or more solubilizing agents. As used herein, the term "solvent" refers to a solvent that can improve solubility or enhance solubility. The term "solvent" as used herein does not include surfactants. In some embodiments, examples of solubilizers include, but are not limited to, glycerin, mannitol, propylene glycol, and combinations thereof. Based on the total volume of the stable pharmaceutical composition, the concentration of the solubilizer may be about 0 mg / mL to about 40 mg / mL, about 0 mg / mL to about 50 mg / mL, about 10 mg / mL to about 50 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, or about 45 mg / mL. In some embodiments, the solubilizer is glycerin. Based on the total volume of the stable pharmaceutical composition, in some embodiments, the solubilizer is glycerin or propylene glycol, with concentrations of approximately 0 mg / mL to approximately 40 mg / mL, approximately 10 mg / mL to approximately 25 mg / mL, approximately 12 mg / mL, approximately 14 mg / mL, approximately 16 mg / mL, approximately 17 mg / mL, approximately 18 mg / mL, approximately 20 mg / mL, approximately 22 mg / mL, or approximately 24 mg / mL. Based on the total volume of the stable pharmaceutical composition, in some embodiments, the solubilizer is mannitol, with concentrations of approximately 0 mg / mL to approximately 50 mg / mL, 0 mg / mL to approximately 40 mg / mL, approximately 10 mg / mL to approximately 25 mg / mL, approximately 12 mg / mL, approximately 14 mg / mL, approximately 16 mg / mL, approximately 17 mg / mL, approximately 18 mg / mL, approximately 20 mg / mL, approximately 22 mg / mL, or approximately 24 mg / mL.

[0054] In some embodiments, the stable pharmaceutical composition comprises one or more osmotic pressure regulators, for example, but not limited to, glycerin, mannitol, propylene glycol, and combinations thereof. In some embodiments, the stable pharmaceutical composition is an isotonic system with the plasma of the subject, and the osmotic pressure of the plasma is typically about 280 mOsm / L to about 320 mOsm / L. In some embodiments, the osmotic pressure of the stable pharmaceutical composition is approximately 280 mOsm / L to approximately 320 mOsm / L, approximately 270 mOsm / L to approximately 330 mOsm / L, or approximately 260 mOsm / L to approximately 340 mOsm / L. In some embodiments, the pharmaceutical composition comprises 40 mg / mL of mannitol and 5 mg / mL of propylene glycol as osmotic pressure modifiers. In some embodiments, the stable pharmaceutical composition is an insulin glargine injection composition or an insulin glargine / GLP-1 combination pharmaceutical composition, and the osmotic pressure modifier is glycerin. The concentration of glycerin in the insulin glargine injection composition is 17 mg / mL, and the concentration of glycerin in the combination pharmaceutical composition is 20 mg / mL. Other applicable osmotic pressure modifiers include, but are not limited to, mannitol, glycerol, propylene glycol, and combinations thereof.

[0055] In some embodiments, the stable pharmaceutical composition comprises one or more preservatives. Optionally, the preservative comprises at least one compound selected from phenol compounds, aromatic alcohol compounds, phenoxyethanol, and chlorohydrin compounds. In some embodiments, the preservative comprises at least one compound selected from phenol, m-cresol, benzyl alcohol, phenoxyethanol, or chlorobutanol. In some embodiments, the preservative is phenol. In some embodiments, the stable pharmaceutical composition has concentrations of approximately 2.0 mg / mL to approximately 4.0 mg / mL, approximately 2.0 mg / mL, approximately 2.1 mg / mL, approximately 2.2 mg / mL, approximately 2.3 mg / mL, approximately 2.4 mg / mL, approximately 2.5 mg / mL, approximately 2.6 mg / mL, approximately 2.7 mg / mL, approximately 2.8 mg / mL, approximately 2.9 mg / mL, approximately 3.0 mg / mL, approximately 3.1 mg / mL, approximately 3.2 mg / mL, approximately 3.3 mg / mL, approximately 3.4 mg / mL, approximately 3.5 mg / mL, approximately 3.6 mg / mL, approximately 3.7 mg / mL, approximately 3.8 mg / mL, approximately 3.9 mg / mL, approximately 4.0 mg / mL, approximately 2.10 mg / mL, approximately 2.11 mg / mL, approximately 2.12 mg / mL, approximately 2.13 mg / mL, approximately 2.14 mg / mL, and approximately 2.15 mg / mL. Contains phenol in mg / mL, approximately 2.16 mg / mL, approximately 2.17 mg / mL, approximately 2.18 mg / mL, approximately 2.19 mg / mL, approximately 2.20 mg / mL, approximately 2.21 mg / mL, approximately 2.22 mg / mL, approximately 2.23 mg / mL, approximately 2.24 mg / mL, approximately 2.25 mg / mL, approximately 2.26 mg / mL, approximately 2.27 mg / mL, approximately 2.28 mg / mL, approximately 2.29 mg / mL, approximately 2.30 mg / mL, approximately 2.31 mg / mL, approximately 2.32 mg / mL, approximately 2.33 mg / mL, approximately 2.34 mg / mL, approximately 2.35 mg / mL, approximately 2.36 mg / mL, approximately 2.37 mg / mL, approximately 2.38 mg / mL, approximately 2.39 mg / mL, or approximately 2.40 mg / mL.

[0056] In some embodiments, the stable pharmaceutical composition has concentrations of approximately 0.1 mg / mL to approximately 10 mg / mL, approximately 0.1 mg / mL to approximately 2.5 mg / mL, approximately 0.1 mg / mL to approximately 1 mg / mL, approximately 0.1 mg / mL, approximately 0.2 mg / mL, approximately 0.3 mg / mL, approximately 0.4 mg / mL, approximately 0.5 mg / mL, approximately 0.6 mg / mL, approximately 0.7 mg / mL, approximately 0.8 mg / mL, approximately 0.9 mg / mL, approximately 1.0 mg / mL, approximately 1.1 mg / mL, approximately 1.2 mg / mL, approximately 1.3 mg / mL, approximately 1.4 mg / mL, approximately 1.5 mg / mL, approximately 1.6 mg / mL, approximately 1.7 mg / mL, approximately 1.8 mg / mL, approximately 1.9 mg / mL, approximately 2.0 mg / mL, approximately 2.1 mg / mL, approximately 2.2 mg / mL, approximately 2.3 mg / mL, and approximately 2.4 mg / mL. It may also contain a GLP-1 receptor agonist in a dose of mg / mL, approximately 2.5 mg / mL, approximately 2.6 mg / mL, approximately 2.7 mg / mL, approximately 2.8 mg / mL, approximately 2.9 mg / mL, approximately 3.0 mg / mL, approximately 3.1 mg / mL, approximately 3.2 mg / mL, approximately 3.3 mg / mL, approximately 3.4 mg / mL, approximately 3.5 mg / mL, approximately 3.6 mg / mL, approximately 3.7 mg / mL, approximately 3.8 mg / mL, approximately 3.9 mg / mL, or approximately 4.0 mg / mL.

[0057] In some embodiments of the stable pharmaceutical compositions disclosed herein, no foreign matter formation was observed in accelerated stability tests (25C3M) at 25°C for 3 months, and the peptide percentage (i.e., the amount of peptide before testing divided by the amount of peptide after testing) was about 93% or higher, preferably about 94% or higher. Examples of corresponding methods are described in the General Methods section of the Examples section.

[0058] Another aspect of the present invention relates to a method for improving the storage stability of a pharmaceutical composition comprising a first active ingredient which includes one or more GLP-1 receptor agonists. In some embodiments of the methods disclosed herein, the method includes the step of mixing one or more surfactants (e.g., alkyl glycosides (e.g., DDMs, but not limited to)) with the pharmaceutical composition. In some embodiments, the pharmaceutical composition is a combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog. Examples of insulin analogs include, but are not limited to, insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin degludec, and insulin detemir.

[0059] In some embodiments, the one or more alkyl glycosides are DDM, sucrose monododecanoate, n-dodecyl β-D-maltoside, n-hexyl-β-D-glucoside, n-heptyl-β-D-glucoside, n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-glucoside, 3-cyclohexyl-1-propyl-β-D-glucoside, n-hexyl-β-D-glucoside It comprises at least one selected from lucopyranoside, n-octyl-β-D-maltoside, n-nonyl-β-D-maltoside, n-decyl-β-D-maltoside, cyclohexyl-methyl-β-D-maltoside, 2-cyclohexyl-ethyl-β-D-maltoside, 3-cyclohexyl-propyl-β-D-maltoside, 4-cyclohexyl-butyl-β-D-maltoside, or 5-cyclohexyl-amyl-β-D-maltoside. In some embodiments, the alkyl glycoside is DDM.

[0060] Another aspect of the present invention relates to the use of a stable pharmaceutical composition in the manufacture of pharmaceuticals used in the following applications: (i) prevention and / or treatment of diabetes mellitus and / or reduction of HbA1c; (ii) delay or prevention of the progression of diabetes mellitus, delay of progression from impaired glucose tolerance to type 2 diabetes mellitus requiring insulin, delay or prevention of insulin resistance and / or delay of progression from type 2 diabetes mellitus that does not require insulin to type 2 diabetes mellitus that requires insulin; (iii) improvement of β-cell function and / or restoration of glucose sensitivity of β-cells; (iv) prevention and / or treatment of eating disorders, decreased gastric motility, and delayed gastric emptying; (v) prevention and / or treatment of complications associated with obesity; (vi) prevention and / or treatment of complications associated with diabetes mellitus; and / or (vii) prevention and / or treatment of cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient.

[0061] Another aspect of the present invention relates to the use of a stable pharmaceutical composition for the following purposes: (i) prevention and / or treatment of diabetes mellitus, and / or reduction of HbA1c; (ii) delay or prevention of the progression of diabetes mellitus, delay of progression from impaired glucose tolerance to type 2 diabetes mellitus requiring insulin, delay or prevention of insulin resistance, and / or delay of progression from type 2 diabetes mellitus that does not require insulin to type 2 diabetes mellitus that requires insulin; (iii) improvement of β-cell function and / or restoration of glucose sensitivity of β-cells; (iv) prevention and / or treatment of eating disorders, decreased gastric motility, and delayed gastric emptying; (v) prevention and / or treatment of complications associated with obesity; (vi) prevention and / or treatment of complications associated with diabetes mellitus; and / or (vii) prevention and / or treatment of cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient.

[0062] Another aspect of the present invention is a method for administering a therapeutically effective amount of a stable pharmaceutical composition to a subject, which is used for the following purposes: (i) prevention and / or treatment of diabetes mellitus, and / or reduction of HbA1c; (ii) delay or prevention of the progression of diabetes mellitus, delay of the progression from impaired glucose tolerance to type 2 diabetes mellitus requiring insulin, delay or prevention of insulin resistance, and / or delay of the progression from type 2 diabetes mellitus that does not require insulin to type 2 diabetes mellitus that requires insulin; (iii) improvement of β-cell function and / or restoration of glucose sensitivity of β-cells; (iv) prevention and / or treatment of eating disorders, decreased gastric motility, and delayed gastric emptying; (v) prevention and / or treatment of complications associated with obesity; (vi) prevention and / or treatment of complications associated with diabetes mellitus; and / or (vii) prevention and / or treatment of cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising insulin, an insulin analog, and / or a derivative of an insulin analog as a second active ingredient.

[0063] Examples are given below to more clearly illustrate the present invention and the embodiments described herein, but these examples should not be construed as limiting the scope of the invention. Unless otherwise noted, the specific materials listed in the following examples are merely illustrative examples for illustrative purposes and are not intended to limit the scope of this disclosure. Those skilled in the art will see that various equivalents, modifications, and alterations can be made without departing from the scope of the invention, and it will be understood that such equivalent embodiments are also included herein. Furthermore, all references made herein are incorporated herein as if they were entirely contained herein. [Examples]

[0064] General method Unless otherwise specified, the methods described in the "General Methods" section of this specification were used in the examples described in the "Examples" section of this specification.

[0065] I. ThT Exam The ThT test is a common method for monitoring protein fibril formation. Thioflavin T (ThT) is a benzothiazole fluorescent dye that binds to amyloid fibrils with high specificity and exhibits enhanced fluorescence after binding. In this test, the kinetic process of protein fibril formation is measured using changes in fluorescence values. When the fluorescence value exceeds a predetermined threshold, fibril formation is determined to have occurred. Unless otherwise specified, in the embodiments of this disclosure, the predetermined threshold is set to three times the initial fluorescence value at the measurement point, i.e., Tn / T1 = 3.

[0066] ThT Exam Procedure: 1. Sample preparation (following the ThT test sample formulation list): Mix the test sample (500 μL) with thioflavin T aqueous solution (1 mM, 25 μL) to obtain the first mixture. Dispense 200 μL of the first mixture into each well of a black V-bottom 96-well plate (Eppendorf), and set up two repeat wells for each sample (dispense each sample into two wells for measurement).

[0067] 2. Incubation: To prevent evaporation and contamination of the sample, the 96-well plate is sealed with sealing tape and placed in a mixer (Thermo Mixer C, Eppendorf) and incubated at 37 °C and 300 rpm under light-shielded conditions. The 96-well plate used here is a 96-well Costar blk / clrbtm (Costar, a laboratory 96-well plate with a black bottom and a transparent top), but other suitable 96-well plates may be used.

[0068] 3. Fluorescence Measurement Conditions: The M5e multifunction microplate reader (Molecular Devices, LLC) was set to fluorescence measurement mode, with the excitation wavelength set to 430 nm, the fluorescence wavelength to 470 nm, and the measurement temperature set to room temperature (approximately 25 °C). Measurements were performed using a 96-well Costar blk / clrbtm plate.

[0069] 4. Sample Measurement: After incubating the test sample for a specified time, fluorescence measurements were performed, and the average of the fluorescence measurements from two repeating wells was used for data analysis. The initial fluorescence value was the fluorescence measurement taken after incubation at 37 °C for 1 hour. After measurement, the 96-well plate was removed, wrapped in aluminum foil, and stored in a light-shielded place.

[0070] II. Stability Test Stability evaluation consisted of two parts: visual inspection and peptide quantity testing. In the visual inspection, stability was confirmed by visually observing the transparency of the test composition and the presence or absence of visible foreign matter. In the peptide quantity testing, the stability of peptides in the composition was evaluated by measuring the amount of peptides using HPLC. Compositions that passed the visual inspection were selected and subjected to accelerated stability testing in a stability test chamber at 25°C, and peptide quantity testing was performed at predetermined measurement points.

[0071] 1. Visual inspection Samples were prepared according to the requirements of the stability test formulation sheet and stored in a stability test chamber at 25°C for accelerated testing. Samples were taken out at various time points (e.g., 14 days, 1 month, 2 months, 3 months, etc.) and visually inspected. Visual inspection confirmed indicators such as the transparency of the composition and the presence or absence of visible foreign matter formation. Compositions that showed no obvious abnormalities in the visual inspection were judged to meet the requirements and were subjected to the next step of HPLC measurement. On the other hand, compositions that showed abnormalities in the visual inspection were judged to not meet the requirements and were not subjected to subsequent measurement steps.

[0072] 2. Peptide quantity test The compositions used for stability testing were stored in a stability test chamber at 25°C and subjected to accelerated stability testing. At various time points (e.g., 14 days, 1 month, 2 months, 3 months, etc.), peptide content testing was performed on compositions that passed visual inspection. 100 μL of each composition was taken as a test sample. The amounts of GLP-1 and insulin glargine in the compositions were measured by HPLC to evaluate their stability.

[0073] The chromatography column used for HPLC was a Thermo BioBasic 18 column (250 × 4.6 mm, 5 μm), with a detection wavelength of 214 nm and a column temperature of 35 °C. Mobile phase A was phosphate buffer (pH 2.5)-sodium chloride-25% acetonitrile, and mobile phase B was phosphate buffer (pH 2.5)-sodium chloride-65% acetonitrile. For gradient elution, a gradient elution program was used in which the concentration of mobile phase B was gradually increased from 4% to 17% over 20 minutes, and then further increased to 53% over 18 minutes.

[0074] Under the same chromatographic conditions as described above, chromatograms can be obtained in which insulin glargine and GLP-1 are separated as individual components.

[0075] By comparing the chromatogram of the combination composition with the chromatograms of insulin glargine alone and GLP-1 alone, the peaks corresponding to insulin glargine, GLP-1, or their derivatives can be identified. Generally, insulin glargine and / or its derivatives are relatively weakly retained in C18 chromatography columns, and their peaks appear in the first half of the chromatogram of the combination drug composition. GLP-1 and / or its derivative, bainaglutide, are relatively strongly retained, and their peaks appear in the second half of the chromatogram of the drug composition. The amount of residual GLP-1 peptide (hereinafter also referred to as "GLP-1 peptide%" or "peptide%" unless otherwise specified) can be determined by calculating the GLP-1 peak area at each stability evaluation point as a ratio to the initial value. In other words, a sample of the composition is taken at each stability evaluation point, analyzed by HPLC, and the peak area corresponding to the GLP-1 chromatographic peak is obtained. Subsequently, the GLP-1 peak area at each stability evaluation point is compared with the GLP-1 peak area at the initial point (t0) to determine the remaining percentage of the GLP-1 peptide. A similar method can be used to calculate the peptide percentage of insulin glargine.

[0076] [Example 1] A series of pharmaceutical compositions (compositions 1-28) containing GLP-1 receptor agonists were prepared using the various components shown in Table 1. Zinc acetate was used as the zinc ion source. ThT tests were performed on each of these compositions, and the results showed that DDM significantly suppressed the formation of GLP-1 fibrillation and improved the physical stability of the pharmaceutical compositions. The results will be described later with reference to Figures 1-8.

[0077] [Table 1-1] [Table 1-2]

[0078] Summary of ThT test results for compositions 1-28 1. The ThT test results for compositions 1-4 (Figure 1) showed that, in the test samples of the insulin glargine / bainaglutide stock solution combination composition, DDM was significantly more effective than other surfactants (e.g., Tween 20 and Tween 80) in suppressing GLP-1 fibrillation formation and improving physical stability. As shown in Figure 1, the increase in fluorescence was not significant before 30 hours (initial stage), and no clear trend of fibrillation was observed. When the incubation time exceeded 30 hours, a clear increase in fluorescence values ​​was observed in composition 1 (solid triangular marker, B+G_phenol) which does not contain surfactants, composition 2 (hollow circular marker, B+G_phenol_0.05%TW20) which contains Tween 20, and composition 3 (solid square marker, B+G_phenol_0.05%TW80) which contains Tween 80. However, no significant increase in fluorescence values ​​was observed in composition 4 (hollow diamond marker, B+G_phenol_0.05%DDM) which contains DDM. The fluorescence values ​​of compositions 1-3 increased rapidly over time, indicating significant protein fiber formation in these compositions. In contrast, no significant change in fluorescence values ​​was observed in the test sample containing DDM until nearly 50 hours. Therefore, in the test sample of the insulin glargine / bainaglutide stock solution combination composition, DDM was shown to be significantly superior to the other surfactants tested in suppressing GLP-1 fibrillation formation and improving physical stability.

[0079] 2. From the ThT test results for compositions 5-8 (Figure 2), it was shown that DDM was significantly more effective than other surfactants (e.g., Tween 20 and Tween 80) in suppressing GLP-1 fibrillation and improving physical stability in the bainaglutide stock solution test samples. As shown in Figure 2, the increase in fluorescence was not significant before 30 hours (initial stage), and no clear trend of fibrillation was observed. When the incubation time exceeded 30 hours, a clear increase in fluorescence values ​​was observed in composition 5 (solid triangular marker, GLP-1_phenol) which does not contain surfactants, composition 6 (hollow circular marker, GLP-1_phenol_0.05%TW20) which contains Tween 20, and composition 7 (solid square marker, GLP-1_phenol_0.05%TW80) which contains Tween 80. However, no significant increase in fluorescence values ​​was observed in composition 8 (hollow diamond marker, GLP-1_phenol_0.05%DDM) which contains DDM. The fluorescence values ​​of compositions 5-7 continued to increase rapidly over time, indicating significant protein fiber formation in these compositions. However, in the test samples containing DDM, no significant change in fluorescence values ​​was observed until nearly 50 hours. Therefore, in the baynaglutide stock solution test samples, DDM was shown to be significantly superior to the other surfactants tested in suppressing GLP-1 fibrillation formation and improving physical stability.

[0080] 3. The ThT test results for compositions 9-12 (Figure 3) showed that DDM significantly improved the physical stability of the bainaglutide composition test samples by suppressing the formation of GLP-1 fibrillation. As shown in Figure 3, the increase in fluorescence was not significant before 30 hours (initial stage), and no clear trend of fibrillation was observed. When the incubation time exceeded 30 hours, a clear increase in fluorescence values ​​was observed in composition 9 (solid square marker, BNLT_Lot1(New)) which did not contain surfactants. However, in compositions containing DDM, no significant increase in fluorescence values ​​was observed even in composition 10 (hollow circle marker, BNLT_Lot2+0.01%DDM) with a low DDM concentration of 0.01%, and similarly, no significant increase in fluorescence values ​​was observed in composition 11 (solid triangular marker, BNLT_Lot2+0.05%DDM) with a DDM concentration of 0.05% and composition 12 (hollow diamond marker, BNLT_Lot2+0.1%DDM) with a DDM concentration of 0.1%. In the test samples containing DDM, no significant change in fluorescence values ​​was observed up to nearly 50 hours. Therefore, in the bainaglutide composition test samples, DDM significantly suppressed the formation of GLP-1 fibrillation and improved physical stability.

[0081] 4. ThT test results for compositions 13-17 (Figure 4) showed that 0.05% DDM significantly suppressed the formation of protein fibrillation and improved physical stability in compositions containing various preservatives. As shown in Figure 4, in the test samples of insulin glargine / bainaglutide stock solution combination compositions containing various preservatives, namely composition 13 containing phenol (solid square marker, B+G_phenol+0.05%DDM), composition 14 containing m-cresol (hollow circle marker, B+G_m-cresol+0.05%DDM), composition 15 containing benzyl alcohol (solid triangular marker, B+G_benzyl alcohol+0.05%DDM), composition 16 containing trichlorobutanol (hollow diamond marker, B+G_trichlorobutanol+0.05%DDM), and composition 17 containing phenoxyethanol (checkerboard square marker, B+G_phenoxyethanol+0.05%DDM), no significant increase in fluorescence was observed throughout the entire test period (over 150 hours). Therefore, it was shown that DDM significantly suppresses the formation of GLP-1 fibrillation and improves physical stability even in compositions containing various preservatives.

[0082] 5. ThT test results for compositions 18-22 (Figure 5) showed that compositions containing DDM significantly suppressed protein fibrillation formation and improved physical stability compared to conventional compositions without DDM. As shown in Figure 5, a significant increase in fluorescence values ​​was observed in conventional compositions without DDM: composition 20 (solid square marker, 20201112-3), composition 21 (solid triangular marker, 20201112-4), and composition 22 (hollow circle marker, 20201112-5). In contrast, no significant change in fluorescence values ​​was observed in composition 18 (checkerboard square marker, 20201112-1) and composition 19 (hollow rhombus marker, 20201112-2), both containing DDM. Therefore, DDM was shown to have advantages over conventional compositions without DDM in terms of suppression of protein fibrillation formation and physical stability.

[0083] 6. ThT test results for compositions 23-28 (Figures 6-8) showed that in compositions containing GLP-1, GLP-1 analogs (e.g., 988927), and GLP-1-based GLP-1 / GIP dual agonists (e.g., P13), DDM significantly suppressed the formation of GLP-1 fibrillation and improved physical stability. After 7 days of incubation, the fluorescence values ​​of compositions 23 and 27 increased to more than 100 times their initial values, and the fluorescence value of composition 25 also increased to more than 35 times its initial value. In contrast, no significant changes in fluorescence values ​​were observed throughout the one-week test period for compositions containing 0.01% DDM: Composition 24 (Figure 6, hollow triangular marker, GLP1-DDM), Composition 26 (Figure 7, hollow triangular marker, P13-DDM), and Composition 28 (Figure 8, hollow triangular marker, 988927-DDM). Therefore, DDM significantly suppresses the formation of GLP-1 fibrillation and improves physical stability in compositions containing GLP-1, GLP-1 analogs (e.g., 988927), and GLP-1-based GLP-1 / GIP dual agonists (e.g., P13).

[0084] 7. DDM improved the solubility of GLP-1 in the presence of phenol at neutral pH. The solubility of GLP-1 was evaluated as a dependence on phenol concentration under neutral pH conditions, under conditions without DDM, with 0.2% DDM, and with 0.4% DDM. As shown in Figure 9, the solubility of GLP-1 decreased with increasing phenol concentration. However, under conditions containing DDM, the solubility of GLP-1 was improved compared to the GLP-1 composition without DDM (diamond-shaped marker, no DDM). Furthermore, under conditions with a higher DDM concentration (triangle marker, 0.4% DDM), the solubility of GLP-1 was further improved compared to conditions with a lower DDM concentration (square marker, 0.2% DDM).

[0085] [Example 2] A series of combination compositions containing a GLP-1 receptor agonist and insulin glargine were prepared, and stability tests were conducted on these compositions. The results showed that DDM improved the stability of the compositions.

[0086] Compositions A1 to A20, having the components shown in Table 2A, were prepared, and zinc chloride was used as the zinc ion source. Both visual inspection and peptide quantity analysis were performed to evaluate stability.

[0087] [Table 2A-1] [Table 2A-2]

[0088] Summary of Stability Test Results 1. The stability test results for compositions A1 to A4 are summarized in Table 2B. These data demonstrate that DDM significantly improves the stability of the compositions compared to other surfactants such as Tween 20 and Tween 80.

[0089] [Table 2B]

[0090] As shown in Table 2B, after an accelerated test at 25°C for one month (25C1M), the peptide percentage of composition A1 (Lot20200519), which does not contain surfactants, decreased to 93.6%, and the peptide percentage of composition A2 (Lot20200713 / BEM040+GLP1 1.0_TW20), which contains Tween 20, decreased to 93.84%. After an accelerated test at 5°C for 14 days (25C14D), composition A3 (Lot20200713 / BEM040+GLP1 1.0_TW80), which contains Tween 80, showed gelation and failed the visual inspection. On the other hand, composition A4 (Lot20200928_4#) containing 0.1% DDM maintained a peptide percentage of 94.66% even after accelerated testing (25C3M) at 25°C for 3 months, and also passed visual inspection. Therefore, these test results demonstrate that DDM significantly improves the stability of the composition compared to Tween 20 and Tween 80.

[0091] 2. The stability test results for compositions A5 to A9 are summarized in Table 2C. These data demonstrate that DDM significantly improves the stability of the compositions compared to conventional excipients.

[0092] [Table 2C]

[0093] As shown in Table 2C, compositions A7-A9 consist of various combinations of glycerin, mannitol, and propylene glycol, but do not contain DDM. Lowering the pH of the composition may be advantageous in improving physical stability and avoiding appearance abnormalities. However, when the pH was lowered, the peptide percentage of compositions A7-A9 decreased to approximately 90% after 3 months in an accelerated stability test (25C3M). In contrast, the combined compositions containing DDM (compositions A5 and A6) maintained a peptide percentage of over 94% even after an accelerated stability test at 25°C for 3 months. Therefore, it was demonstrated that DDM significantly improves the stability of the composition compared to conventional excipients.

[0094] 3. The stability test results for compositions A10 to A14 are summarized in Table 2D. These data demonstrate that DDM significantly improves the stability of compositions containing various preservatives.

[0095] [Table 2D]

[0096] The preservatives used in the test were those commonly used in injection-grade compositions, including phenol, m-cresol, benzyl alcohol, trichlorobutanol, and phenoxyethanol. All tested compositions containing DDM showed a peptide percentage exceeding 92% in an accelerated stability test at 25°C for 3 months, and passed the visual inspection. Therefore, it was demonstrated that DDM not only significantly improves the stability of the compositions but is also compatible with various preservatives commonly used in injection formulations.

[0097] 4. The stability test results for compositions A15-A18 are summarized in Table 2E. Compositions with different DDM concentrations were compared.

[0098] [Table 2E]

[0099] Composition A15 (0.05% DDM) failed the visual inspection after an accelerated stability test (25C60D) at 25°C for 2 months. In contrast, all compositions containing 0.1% or more DDM passed the visual inspection after an accelerated stability test (25C90D) at 25°C for 3 months, and all had GLP-1 peptide percentages exceeding 93%. Therefore, it was shown that a DDM concentration of 0.1% or more significantly improves the physical stability of the composition compared to a DDM concentration of approximately 0.05%.

[0100] 5. The stability test results for compositions A16, A19, and A20 are summarized in Table 2F. Compositions containing DDM were compared under different pH conditions.

[0101] [Table 2F]

[0102] As shown in Table 2F, in the pH range of 4.0 to 4.5, the combined composition containing 0.1% DDM did not fail the visual inspection in the accelerated stability test at 25°C for 3 months, and the peptide percentage was above 93% in all cases. Therefore, it was demonstrated that DDM effectively improves the stability of the formulation in the pH range of 4.0 to 4.5.

[0103] 6. A series of combination compositions (compositions A30 to A41) containing a GLP-1 receptor agonist and insulin glargine were prepared, and their stability was evaluated.

[0104] [Table 2G]

[0105] As shown in Table 2H, in the pH range of 3.8 to 4.4, the combination composition containing 0.1% DDM maintained an insulin glargine peptide percentage of over 97.23% and a GLP-1 peptide percentage of over 95.49% after an accelerated stability test at 25°C for 3 months. Therefore, it was demonstrated that DDM effectively improves the stability of the formulation in the pH range of 3.8 to 4.4.

[0106] [Table 2H]

[0107] As shown in Table 2I, in the pH range of 3.8 to 4.4, the combination composition containing 0.1% DDM maintained an insulin glargine peptide percentage of over 97.23% and a GLP-1 peptide percentage of over 95.49% after an accelerated stability test at 25°C for 3 months. Therefore, it was demonstrated that DDM effectively improves the stability of the formulation in the pH range of 3.8 to 4.4.

[0108] [Table 2I]

[0109] Combination compositions containing solubilizers consisting of different combinations of glycerin / mannitol / propylene glycol were tested under conditions with and without DDM. The test combination compositions without DM failed the visual inspection after the accelerated 40C28D test. On the other hand, when 0.1% DDM was included in the combination composition, the appearance after the accelerated 40C28D test improved, and the GLP-1 peptide percentage after the accelerated 40C14D and 40C28D tests was over 93% and over 87%, respectively.

[0110] [Example 3] A series of combination compositions containing a GLP-1 receptor agonist and insulin aspart were prepared, and stability tests were conducted on these compositions. The results showed that DDM improved the stability of the compositions. Unless otherwise specified, the buffering system for the combination compositions in Example 3 was PB.

[0111] Compositions B1 to B20, containing the components shown in Table 3A, were prepared, and both visual inspection and peptide quantity testing were performed to evaluate their stability.

[0112] [Table 3A]

[0113] 1. Compositions B1-B5: These are embodiments of a combination pharmaceutical composition of insulin aspart and GLP-1 (veinaglutide), with varying NaCl concentrations.

[0114] To show the size of the particles present in compositions B1 to B5, DLS (dynamic light scattering) light intensity spectra were used.

[0115] In the combination drug composition (composition B1) that did not contain NaCl, after an accelerated test (40C14D) at 40°C for 14 days, large GLP-1 particles exceeding 100 nm became dominant, suggesting that significant protein aggregation of GLP-1 occurred.

[0116] In the combined pharmaceutical composition (composition B2) with a NaCl concentration of 10 mM, the amount of large particles exceeding 100 nm was significantly reduced.

[0117] In the combination pharmaceutical composition with an NaCl concentration of 20 mM (composition B3) and combination pharmaceutical compositions with higher NaCl concentrations (compositions B4-B5), no large particles exceeding 100 nm were observed, and the majority of GLP-1 particles had a particle size of less than 10 nm.

[0118] The stability test results for compositions B1 to B5 are summarized in Table 3B below.

[0119] [Table 3B(1)]

[0120] The combination composition without NaCl failed the visual inspection after the accelerated 40C14D test. As the NaCl concentration exceeded 10 mM, the appearance of the combination composition improved significantly during the accelerated 40C14D test, with subsequent insulin aspart peptide percentages exceeding 90.8% and GLP-1 peptide percentages exceeding 90.5%.

[0121] 2. Compositions B6-B8: These are embodiments of a combination pharmaceutical composition of insulin aspart and GLP-1 (veinaglutide) with varying DDM concentrations.

[0122] After an accelerated test at 40°C for two months (40C2M), composition B6 (0.16% DDM) failed the visual inspection and showed a white, cottony precipitate.

[0123] Composition B8 (0.25% DDM) failed the visual inspection after an accelerated test at 40°C for 14 days (40C14D), but the same composition passed the visual inspection after being stored at 4°C for 3 months (4C3M).

[0124] Composition B7 (0.2% DDM) passed visual inspection both after an accelerated test at 40°C for 14 days (40C14D) and after storage at 4°C for 3 months (4C3M).

[0125] 3. This is an embodiment of a combination pharmaceutical composition of insulin aspart and GLP-1 (vainaglutide), with compositions B9-B12 having pH values ​​of 7.4, 7.6, and 8.15.

[0126] The stability test results for compositions B1 to B5 are summarized in Table 3B below.

[0127] [Table 3B(2)]

[0128] The combined composition with a pH of 8.15 (composition B9) maintained its appearance stability during storage at 4°C for 3 months. However, peptide percentage data showed that after the 40C2W accelerated trial, the peptide percentage of insulin aspart was lower in composition B9 at pH 8.15 than in composition B10 at pH 7.6.

[0129] Compositions B11 (pH 7.6) and B12 (pH 7.4) both passed the visual inspection after the accelerated 40C2W test. However, after the accelerated 40C4W test, although both compositions maintained good fluidity, the presence of foreign matter was detected.

[0130] 4. Compositions B13-B16: Embodiments of combination pharmaceutical compositions of insulin aspart and GLP-1 (vainaglutide) with varying Zn ion concentrations (1 μg / mL, 10 μg / mL, 20 μg / mL, and 25 μg / mL).

[0131] The stability test results for compositions B13 to B16 are summarized in Table 3C below.

[0132] [Table 3C]

[0133] Composition B13 contains EDTA that can form chelates with all Zn in the solution; therefore, composition B13 can be considered a composition that does not contain Zn (zinc) ions. The combined composition that does not contain Zn ions (composition B13) and combined compositions with various Zn ion concentrations (compositions B14-B16) were examined for appearance and turbidity after storage at 4 °C / 25 °C / 40 °C. The tested compositions either formed precipitates or showed turbidity exceeding the predetermined value (>3.0) after accelerated testing at 25 °C and 40 °C. Furthermore, the tested compositions also showed turbidity exceeding the predetermined value (>1.5) after accelerated testing at 4 °C.

[0134] [Example 4] The pharmacokinetics (PK) of one embodiment of a combination pharmaceutical composition of insulin aspart and GLP-1 (vainaglutide) were investigated in beagle dogs (male, 2-3 years old, 9-11 kg).

[0135] Eleven beagle dogs were acclimatized to the laboratory environment, then fasted for 20 hours (without drinking water), and fed 4 hours after administration. The beagle dogs were randomly divided into three groups (n=3) based on body weight: a GLP-1 monotherapy group (20 μg / kg, 50 μL / kg), an insulin aspart monotherapy group (0.5 U / kg, 50 μL / kg), and a group receiving a combination of GLP-1 (10 μg / kg) and insulin aspart (0.5 U / kg) (50 μL / kg).

[0136] Before and after administration, 1.0 mL of whole blood was rapidly collected using a 1 mL syringe that had been pre-rinsed with 1 mL each of a DPPIV inhibitor and a protease inhibitor. In the GLP-1 alone group, blood samples were collected at 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, and 120 minutes after administration. In the insulin aspart alone group and the combined group, blood samples were collected at 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 120, 180, 240, 300, and 480 minutes after administration. The collected whole blood was placed in an anticoagulation tube containing EDTA-K2. In the EDTA-K2 anticoagulation tube, 10 μL of a DPPIV inhibitor and 10 μL of a protease inhibitor had been added in advance. The obtained mixture was kept in an ice bath within 30 minutes before centrifugation and then centrifuged at 4 °C, 300 g for 5 minutes to collect plasma. 0.3 mL and 0.1 mL of the obtained plasma were separately aliquoted into test tubes, stored at -80 °C, and then used for the measurement of active GLP-1 and insulin aspart, respectively.

[0137] The concentrations of insulin aspart and GLP-1 at each blood sampling time point were measured using an ELISA kit. Pharmacokinetic parameters were calculated by compartmental analysis based on an in vitro administration model (two-compartment model) for plasma data using PKSolver 2.0.

[0138] The time course of the insulin aspart concentration is shown in Figure 10, and the PK parameters are summarized in Table 4A.

[0139]

Table 4A

[0140] The T of the insulin aspart alone group max was approximately 45 minutes, and the T of the combined group maxThe time to reach peak blood concentration after administration was approximately 32 minutes. In other words, the time to reach peak blood concentration after administration in combination therapy was 13 minutes shorter compared to insulin aspart monotherapy, suggesting that insulin aspart has a faster onset of action in combination therapy than in monotherapy. max and AUC 0-t These levels were 1.22 times and 1.29 times higher, respectively, compared to insulin aspart monotherapy. Similarly, GLP-1 concentrations were measured and analyzed at each blood sampling time in the GLP-1 monotherapy group and the combination therapy group. The time course of GLP-1 concentration is shown in Figure 11. Considering that the GLP-1 dose in the GLP-1 monotherapy group was twice that of the combination therapy group, the PK parameters were corrected to those of the same GLP-1 dose (10 μg / kg). The corrected PK parameters are summarized in Table 4B.

[0141] [Table 4B]

[0142] GLP-1 T in the GLP-1 monotherapy group max The time interval was approximately 17 minutes, and the T15 time for GLP-1 in the combination group was approximately 17 minutes. max The time to reach peak concentration after administration was approximately 14 minutes. The time to reach peak concentration after administration was slightly shorter in the combination group compared to the GLP-1 monotherapy group. The corrected C in the combination group was max and AUC 0-t These levels were 1.24 times and 1.09 times, respectively, compared to the GLP-1 monotherapy group.

[0143] As GLP-1, we used baynaglutide (2 mg / mL, 10 mM acetate, 30 mg / mL mannitol, 2.2 mg / mL phenol, 5 mg / mL propylene glycol, pH 4.0, manufactured by Shanghai Benemae).

[0144] The insulin aspart preparation used was one containing insulin aspart 100 U (3.5 mg) / mL, glycerol 16 mg / mL, phenol 1.5 mg / mL, m-cresol 1.72 mg / mL, sodium dihydrogen phosphate dihydrate 1.25 mg / mL, sodium chloride 0.58 mg / mL, zinc 19.6 μg / mL, and pH 7.4 (manufactured by Novo Nordisk).

[0145] The combined composition contained insulin aspart 50 U / mL, GLP-1 1.0 mg / mL, glycerol 20 mg / mL, phenol 3.2 mg / mL, zinc 20 μg / mL, pH 7.6, DDM 0.2%, NaCl 40 mM, and phosphate buffer (PB).

[0146] In this specification, when terms such as "one embodiment," "several embodiments," "example," "specific example," or "several examples" are used, it means that specific features, structures, materials, or properties described in relation to such embodiment or example are included in at least one embodiment or example of the present invention.

[0147] The schematic representations of the terms used herein do not necessarily refer to the same embodiments or examples. Furthermore, the specific features, structures, materials, or properties described may be used in any combination in any manner as appropriate in one or more embodiments or examples.

[0148] While embodiments of this disclosure have been shown and described above, it should be understood that these embodiments are illustrative and should not be construed as limiting this disclosure. It will be obvious to those skilled in the art that various changes, modifications, substitutions, and alterations can be made to the above embodiments within the scope of this disclosure without departing from the principles and purposes of this disclosure.

Claims

1. A pharmaceutical composition comprising a first active ingredient containing a GLP-1 receptor agonist, and one or more surfactants containing one or more alkyl glycosides.

2. A pharmaceutical composition according to claim 1, further comprising a second active ingredient comprising one or more of insulin, an insulin analog, and a derivative of an insulin analog, wherein optionally, the insulin analog comprises at least one selected from insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin degludec, or insulin detemir. The pharmaceutical composition optionally contains 0.1 to 1% by mass of insulin, an insulin analog, or a derivative of an insulin analog, based on its total mass, and optionally contains 0.1 to 0.4% by mass of the insulin, insulin analog, or derivative of an insulin analog.

3. A pharmaceutical composition according to claim 1, wherein the alkyl glycoside has a structure in which a sugar moiety and an alkyl moiety are linked by a linking group, The aforementioned sugar moiety comprises at least one sugar selected from glucose, maltose, sucrose, or trehalose. The alkyl group contains approximately 10 to 16 carbon atoms. A pharmaceutical composition comprising an alkyl glycoside, wherein the linking group comprises at least one bond selected from a glycosidic bond, a thioglycosidic bond, or an amide bond.

4. A pharmaceutical composition according to claim 3, wherein the alkyl chain of the alkyl moiety comprises 10 to 14 carbon atoms.

5. A pharmaceutical composition according to claim 3, wherein the sugar moiety comprises maltose or sucrose.

6. The pharmaceutical composition according to claim 1, wherein the alkyl glycoside is n-dodecyl β-D-maltoside (DDM), sucrose monododecanoate, n-dodecyl β-D-maltoside, n-hexyl-β-D-glucoside, n-heptyl-β-D-glucoside, n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-glucoside, 3-cyclohexyl-1-propyl-β-D-glucoside, n-hexyl-β-D-glucopyranoside, n-octyl-β-D-maltoside, n-nonyl-β- The compound comprises at least one compound selected from D-maltoside, n-decyl-β-D-maltoside, cyclohexyl-methyl-β-D-maltoside, 2-cyclohexyl-ethyl-β-D-maltoside, 3-cyclohexyl-propyl-β-D-maltoside, 4-cyclohexyl-butyl-β-D-maltoside, or 5-cyclohexyl-amyl-β-D-maltoside. A pharmaceutical composition wherein, optionally, the concentration of the alkyl glycoside is approximately 0.05 to approximately 0.50% by mass or approximately 0.10 to approximately 0.20% by mass, based on the total mass of the pharmaceutical composition.

7. A pharmaceutical composition according to claim 1, wherein the GLP-1 receptor agonist is a peptide agonist.

8. A pharmaceutical composition according to claim 7, wherein the GLP-1 receptor agonist is also a GIP agonist.

9. A pharmaceutical composition according to claim 7, wherein the GLP-1 receptor agonist comprises at least one of the following: GLP-1, GLP-1 analog, or derivatives of GLP-1 analog; Extended GLP-1 fragments, their analogues or derivatives; and Severed GLP-1 fragments, their analogues, or derivatives thereof.

10. A pharmaceutical composition according to claim 7, wherein the GLP-1 analog comprises exendin-4.

11. A pharmaceutical composition according to claim 1, wherein the GLP-1 receptor agonist comprises at least one selected from liraglutide, exenatide, lixisenatide, albiglutide, veinaglutide, dulaglutide, semaglutide, pepamotidide, and tilpotide.

12. A pharmaceutical composition according to claim 1, wherein the first active ingredient comprises or is veinaglutide, and the second active ingredient comprises or is insulin glargine.

13. A pharmaceutical composition according to claim 1, wherein the first active ingredient comprises or is veinaglutide, and the second active ingredient comprises or is insulin aspart.

14. A pharmaceutical composition according to any one of claims 1 to 12, wherein the pharmaceutical composition is a liquid pharmaceutical composition or a premix aqueous pharmaceutical composition for injection.

15. A pharmaceutical composition according to claim 14, further comprising a preservative, wherein the preservative comprises at least one compound selected from phenolic compounds, aromatic alcohol compounds, phenoxyethanol, or chlorhydrin compounds.

16. A pharmaceutical composition according to claim 15, wherein the preservative comprises at least one of phenol, m-cresol, benzyl alcohol, phenoxyethanol, or chlorobutanol.

17. A pharmaceutical composition according to claim 15, wherein the concentration of the GLP-1 receptor agonist is about 0.1 mg / mL to about 10 mg / mL.

18. A pharmaceutical composition according to claim 15, wherein the pH of the pharmaceutical composition is about 3.0 to about 9.

0. Optionally, the insulin analog is insulin glargine, the pH of the pharmaceutical composition is approximately 3.0 to approximately 5.0, and further optionally, the pH is approximately 4.0 to approximately 4.

5. A pharmaceutical composition wherein, optionally, the insulin analog is insulin aspart, and the pH of the pharmaceutical composition is approximately 7.0 to approximately 9.

0.

19. A pharmaceutical composition according to claim 15, further comprising Zn ions, wherein the concentration of Zn ions is optionally about 1 μg / mL to about 100 μg / mL, about 10 μg / mL to about 50 μg / mL, or about 20 μg / mL to about 40 μg / mL.

20. A pharmaceutical composition according to claim 1, wherein the pharmaceutical composition further comprises a solubilizer, the solubilizer comprising at least one selected from glycerin, mannitol, propylene glycol, and combinations thereof, and the concentration of the solubilizer is about 10 mg / mL to about 50 mg / mL based on the total volume of the pharmaceutical composition, preferably the solubilizer is glycerin, and its concentration is about 10 mg / mL to about 25 mg / mL based on the total volume of the pharmaceutical composition. Alternatively, the pharmaceutical composition may contain none of polysorbate 20, polysorbate 80, poloxamer 188, hydroxypropyl beta-cyclodextrin, or amino acids, and optionally contain only an alkyl glycoside as the surfactant.

21. A pharmaceutical composition according to claim 1, wherein no foreign matter is formed in an accelerated stability test at 25°C for 3 months, and the peptide percentage is approximately 93% or more.

22. A method for improving the storage stability of a pharmaceutical composition containing a GLP-1 receptor agonist, comprising the step of mixing one or more alkyl glycosides with the pharmaceutical composition.

23. A method according to claim 22, wherein the pharmaceutical composition further comprises insulin or an insulin analog or a derivative of an insulin analog, wherein the insulin analog comprises at least one selected from insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin degludec and insulin detemir.

24. The method according to claim 22, wherein the alkyl glycoside is n-dodecyl β-D-maltoside (DDM), sucrose monododecanoate, n-dodecyl β-D-maltoside, n-hexyl-β-D-glucoside, n-heptyl-β-D-glucoside, n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-glucoside, 3-cyclohexyl-1-propyl-β-D-glucoside, n-hexyl-β-D-glucopyranoside, n-octyl-β-D-maltoside, n-nonyl A method comprising at least one of the following: 1-β-D-maltoside, n-decyl-β-D-maltoside, cyclohexyl-methyl-β-D-maltoside, 2-cyclohexyl-ethyl-β-D-maltoside, 3-cyclohexyl-propyl-β-D-maltoside, 4-cyclohexyl-butyl-β-D-maltoside, or 5-cyclohexyl-amyl-β-D-maltoside.

25. Use as a pharmaceutical for at least one of the uses listed below (i) to (vii), wherein the pharmaceutical comprises the pharmaceutical composition described in any one of claims 1 to 20: (i) Prevention and / or treatment of diabetes, and / or reduction of HbA1c; (ii) Delaying or preventing the progression of diabetes mellitus, delaying the progression from impaired glucose tolerance to insulin-dependent type 2 diabetes mellitus, delaying or preventing insulin resistance, and / or delaying the progression from insulin-free type 2 diabetes mellitus to insulin-dependent type 2 diabetes mellitus; (iii) Improvement of β-cell function and / or restoration of glucose sensitivity of β-cells; (iv) Prevention and / or treatment of eating disorders, reduction of gastric motility, and delay of gastric emptying; (v) Prevention and / or treatment of complications of obesity; (vi) Prevention and / or treatment of complications of diabetes; and / or (vii) Prevention and / or treatment of cardiovascular disease.

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