Pharmaceutical composition and use thereof, and method for improving storage stability of pharmaceutical composition

Abstract

Embodiments of stable pharmaceutical compositions, methods for improving the storage stability of pharmaceutical compositions, and uses of the stable pharmaceutical compositions are provided. The pharmaceutical composition comprises a first active ingredient comprising a GLP-1 receptor agonist and a surfactant. In certain embodiments, the surfactant comprises an alkyl glycoside. In certain embodiments, one or more alkyl glycosides (e.g., but not limited to DDM) inhibit the formation of GLP-1 fibrosis in a pharmaceutical composition comprising a GLP-1 receptor agonist, e.g., GLP-1 or an analog thereof, or a derivative of a GLP-1 analog, and thereby improve physical stability.

Description

[0001] Priority Claim The present application claims priority to Chinese Patent Application No. 2023105246295, filed May 10, 2023, the entire contents of which are hereby incorporated by reference in its entirety. TECHNICAL FIELD

[0002] The present application relates to the field of biotechnology / pharmaceuticals, and more specifically, the present application relates to a pharmaceutical composition comprising a GLP-1 receptor agonist and a method for improving the storage stability of the pharmaceutical composition and use of the pharmaceutical composition in the manufacture of a medicament. BACKGROUND

[0003] Approximately 120 million people worldwide suffer from diabetes. Exogenous insulin is mainly used to treat diabetes. Insulin is the only hormone in the body that lowers blood glucose and promotes the synthesis of glycogen, fat and protein. Insulin glargine is an analog of human insulin, which can be obtained by recombinant DNA technology. After insulin glargine is injected into subcutaneous tissue, a small amount of insulin glargine continues to be released by the fine sediment formed by neutralizing the acidic solution, resulting in a predictable, long-acting, smooth and peak-free plasma concentration / time profile.

[0004] Glucagon-like peptide 1 (hereinafter referred to as GLP-1) is an incretin secreted peptide, which has the following physiological effects: glucose concentration-dependent insulin secretion, thereby reducing the risk of hypoglycemia; promoting insulin synthesis in the body, promoting β-cell differentiation generation, inhibiting glucagon release, inhibiting gastric emptying and eating impulse, and increasing sensitivity to insulin receptors. It has been clinically proven to have advantages such as hypoglycemia, weight loss, and increased cardiovascular benefits.

[0005] The combination of insulin and GLP-1 receptor agonists can take full advantage of the complementary clinical advantages of both, such as glucose reduction, weight loss, reduced risk of hypoglycemia, and reduced insulin resistance. Premixed compositions containing basal insulin and GLP-1 are easier to administer, and therefore have a significant market demand.

[0006] However, the current pharmaceutical composition of GLP-1 receptor agonists still has room for improvement. SUMMARY

[0007] One aspect of the present invention relates to a stable pharmaceutical composition comprising a first active ingredient comprising one or more GLP-1 receptor agonists; and one or more surfactants (such as, but not limited to, alkyl glycosides (such as, but not limited to, n-dodecyl beta-D-maltoside (DDM))). Embodiments of the stable pharmaceutical composition disclosed herein exhibit improved stability of the one or more GLP-1 receptor agonists. In certain embodiments, the improved stability relates to physical stability. In certain embodiments, the stable pharmaceutical composition is a stable liquid pharmaceutical composition. In certain embodiments, the stable liquid pharmaceutical composition is a stable aqueous pharmaceutical composition. In certain embodiments, the stable aqueous pharmaceutical composition is a stable aqueous injectable pharmaceutical composition. In certain embodiments, the stable aqueous injectable pharmaceutical composition is a single-dose or multi-dose aqueous injectable pharmaceutical composition. 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.

[0008] In certain embodiments, the stable pharmaceutical composition further comprises a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog, and can be referred to herein as a stable combination pharmaceutical composition. Embodiments of the stable combination pharmaceutical composition disclosed herein also exhibit high stability of the first and second active ingredients. In certain embodiments, the stability relates to physical stability. In certain embodiments, the stable combination pharmaceutical composition is a stable liquid combination pharmaceutical composition. In certain embodiments, the stable liquid combination pharmaceutical composition is a stable aqueous combination pharmaceutical composition. In certain embodiments, the stable aqueous combination pharmaceutical composition is a stable aqueous injectable combination pharmaceutical composition. In certain embodiments, the stable aqueous injectable combination pharmaceutical composition is a single-dose or multi-dose aqueous injectable combination pharmaceutical composition.

[0009] Another aspect of the present invention relates to a method of improving the storage stability of a pharmaceutical composition comprising a first active ingredient comprising one or more GLP-1 receptor agonists. In certain embodiments of the method disclosed herein, the method comprises mixing one or more surfactants (such as, but not limited to, alkyl glycosides (such as, but not limited to, DDM)) with the pharmaceutical composition. In certain 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.

[0010] Another aspect of the invention relates to the use of the stable pharmaceutical composition in the preparation of a medicament for: (i) preventing and / or treating diabetes, and / or lowering HbA1C; (ii) delaying or preventing the progression of diabetes, delaying the progression of impaired glucose tolerance to insulin-dependent type 2 diabetes, delaying or preventing insulin resistance and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes; (iii) improving β-cell function, and / or restoring β-cell glucose sensitivity; (iv) preventing and / or treating eating disorders; decreased gastric motility; delayed gastric emptying; (v) preventing and / or treating complications of obesity; (vi) preventing and / or treating complications of diabetes; and / or (vii) preventing and / or treating cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog.

[0011] Another aspect of the invention relates to the use of stable pharmaceutical compositions for: (i) prevention and / or treatment of diabetes, and / or reduction of HbA1C; (ii) delaying or preventing the progression of diabetes, delaying the progression of impaired glucose tolerance to insulin-dependent type 2 diabetes, delaying or preventing insulin resistance and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes; (iii) improvement of β-cell function, and / or restoration of β-cell glucose sensitivity; (iv) prevention and / or treatment of eating disorders; decreased gastric motility; delayed 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. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog.

[0012] Another aspect of the invention relates to methods for: (i) preventing and / or treating diabetes, and / or lowering HbA1C; (ii) delaying or preventing the progression of diabetes, delaying the progression of impaired glucose tolerance to insulin-dependent type 2 diabetes, delaying or preventing insulin resistance and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes; (iii) improving β-cell function, and / or restoring β-cell glucose sensitivity; (iv) preventing and / or treating eating disorders; decreased gastric motility; delayed gastric emptying; (v) preventing and / or treating complications of obesity; (vi) preventing and / or treating complications of diabetes; and / or (vii) preventing and / or treating cardiovascular disease, comprising administering a therapeutically effective amount of a stable pharmaceutical composition to a subject. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog. Attached Figure Description

[0013] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following accompanying drawings, in which: Figure 1 The ThT test results of an insulin glargine / GLP-1 combination composition according to one embodiment of the present invention are shown. Figure 2 The ThT test results of a benaglutide stock solution according to one embodiment of the present invention are shown. Figure 3 The ThT test results of benaglutide compositions containing and without different concentrations of DDM according to one embodiment of the present invention are shown. Figure 4 The ThT test results of a composition comprising different antibacterial agents of insulin glargine / GLP-1-DDM combinations according to an embodiment of the present invention are shown. Figure 5 The ThT test results of a glargine insulin / GLP-1 combination composition according to one embodiment of the present invention are shown; and Figure 6 The physical stability of GLP-1 pharmaceutical compositions having 0.1% DDM and without DDM according to several embodiments disclosed in this invention is shown.

[0014] Figure 7The physical stability of P13 (peptide sequence: ELAEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2, GLP-1 / GIP dual agonist) pharmaceutical compositions having 0.1% DDM and without DDM, according to several embodiments disclosed in this invention, is shown.

[0015] Figure 8 Several embodiments of the present invention are shown, demonstrating the physical stability of 988927 (peptide sequence: HGEGTFTSDSSSYLEEQAAKEFIAWLVKGRG) with 0.1% DDM and without DDM.

[0016] Figure 9 The relationship between GLP-1 solubility and phenol concentration is shown in the presence or absence of DDM.

[0017] Figure 10 The plasma concentrations of aspart insulin in beagles after administration of compositions containing only aspart insulin and combinations of aspart insulin / GLP-1 according to embodiments disclosed in the present invention are shown.

[0018] Figure 11 The plasma concentrations of GLP-1 in beagles after administration of the GLP-1-only composition and the aspartate insulin / GLP-1 combination composition according to the embodiments disclosed in this invention are shown. Detailed Implementation

[0019] Embodiments of the present invention are described in detail below. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain the present invention, but should not be construed as limiting the present invention.

[0020] definition Unless otherwise stated, the term "active ingredient" as used in this invention refers to a substance or combination of substances that produces a therapeutic or preventive effect in a pharmaceutical composition. For example, according to embodiments of the invention, the active ingredient may be one or more GLP-1 receptor agonists, or insulin or its analogues or derivatives may be additionally added to the pharmaceutical composition so that the pharmaceutical composition can exert the desired therapeutic or preventive effect when administered to a subject.

[0021] Unless otherwise stated, the term "surfactant" as used in this invention refers to a class of chemical substances that can reduce the surface tension and interfacial tension of liquids, typically composed of hydrophobic and hydrophilic groups. Surfactants are commonly used to regulate the solubility, stability, and bioavailability of drugs, thereby affecting drug absorption, distribution, metabolism, and excretion.

[0022] Unless otherwise stated, the term "alkyl glycoside" as used in this invention refers to a type of surfactant composed of a sugar group and an alkyl chain. These surfactants exhibit good surface activity and biodegradability. These compounds are typically obtained by enzymatically catalyzing the reaction of sugars with fatty alcohols under specific temperature and pressure. Because the molecular structure of alkyl glycosides includes both hydrophobic and hydrophilic groups, they possess properties similar to conventional surfactants. Specifically, 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 invention have discovered that alkyl glycosides (e.g., but not limited to DDM) can be used as nonionic surfactants in the preparation of the pharmaceutical compositions of this invention to improve the stability, solubility, and bioavailability of the drug, and can also reduce the toxicity and irritation of the formulation, thereby improving the safety and tolerability of the drug.

[0023] Unless otherwise stated, the term "analogue" as used in this invention refers to a molecule that is structurally similar to the original molecule but has the same or different chemical properties. For example, in drug 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 that alter their pharmacological properties, such as absorption, metabolism, or drug efficacy. These analogs may have better pharmacological or toxicological properties and therefore have the potential for further drug development. More specifically, in this invention, "analog" is used to refer to a peptide in which one or more amino acid residues of the parent peptide are substituted with other amino acid residues and / or in which one or more amino acid residues of the parent peptide are deleted and / or one or more amino acid residues are added to the parent peptide. Such addition typically occurs at the N-terminus or C-terminus of the parent peptide, or both. According to specific embodiments, the "analytes" in this invention are generally peptides in which 6 or fewer amino acids of the parent peptide are substituted and / or added and / or deleted, more preferably peptides in which 3 or fewer amino acids of the parent peptide are substituted and / or added and / or deleted, and most preferably peptides in which one amino acid of the parent peptide is substituted and / or added and / or deleted.

[0024] Unless otherwise stated, the term "derivative" as used in this invention refers to a new molecule obtained from an original molecule through chemical modification or transformation. For example, in drug research, researchers can start with a known drug molecule and synthesize a series of new compounds by altering, substituting, or modifying its structure. These new molecules are called compounds of the drug molecule. Derivatives can have better pharmacological properties, better safety, or fewer side effects, and therefore play an important role in drug development. Specifically, in this invention, "derivative" is used to refer to a peptide in which one or more amino acid residues of the parent peptide have been introduced with substituents. Typical substituents can be amides, sugars, alkyl groups, acyl groups, esters, polyethylene glycol groups, etc.

[0025] Unless otherwise stated, the term "GLP-1 receptor agonist" as used in this invention refers to a class of drugs that help lower blood glucose levels to treat diabetes by mimicking the biological activity of glucagon-like peptide-1 (GLP-1) to promote insulin secretion and inhibit glucose production and release. GLP-1 is a peptide hormone secreted by intestinal L cells and brainstem neurons. It has multiple biological functions, including suppressing appetite, stimulating insulin secretion, and inhibiting glucose production and release. GLP-1 receptor agonists stimulate GLP-1 receptors, enhance insulin secretion, and inhibit glucose production and release. Due to their effectiveness and good tolerability in glycemic control, GLP-1 receptor agonists have become one of the first-line drugs for treating type 2 diabetes. In addition to diabetes treatment, GLP-1 receptor agonists are also being studied in other areas such as obesity, cardiovascular disease, and neurodegenerative diseases.

[0026] Unless otherwise stated, the term "treatment" as used in this invention means the use of a drug to alleviate or cure a disease state, and "prevention" means the use of a drug to prevent the occurrence of a disease or reduce the likelihood of a disease, which may prevent or delay the onset or progression of such diseases.

[0027] Generally, alkyl glycosides (e.g., but not limited to DDM) have been used as mucosal absorption enhancers for peptides, proteins, and small molecules. Surprisingly, in various embodiments of the stable pharmaceutical compositions disclosed in this invention, the presence of alkyl glycosides (e.g., but not limited to DDM) improves 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 in this invention, the stability of one or more active ingredients is sufficiently high that the embodiments can be used in single-dose or multi-dose aqueous injectable compositions.

[0028] Pharmaceutical compositions of GLP-1 receptor agonists, especially their liquid compositions, tend to form fibrous material during storage, which makes obtaining pharmaceutical compositions with long-term stability challenging. As shown in the embodiments provided by the present invention, unexpectedly, certain surfactants 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., but not limited to DDM) improve the stability of GLP-1 receptor agonists in various embodiments of the aqueous pharmaceutical compositions.

[0029] One aspect of the 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.

[0030] In some embodiments, the surfactant includes alkyl glycosides (e.g., but not limited to DDM). Embodiments of the stable pharmaceutical compositions disclosed in this invention demonstrate improved stability of one or more GLP-1 receptor agonists. In some embodiments of the stable pharmaceutical compositions disclosed in this invention, when the GLP-1 receptor agonist comprises, for example, GLP-1 or its analogues or derivatives, the presence of an alkyl glycoside (e.g., but not limited to DDM) can prevent the formation of GLP-1 fibrillation and thereby improve the physical stability of the GLP-1 receptor agonist. Alkyl glycosides (e.g., but not limited to DDM) are significantly superior to other types of surfactants.

[0031] In some embodiments, the improved stability involves 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 injectable pharmaceutical composition. In some embodiments, the stable aqueous injectable pharmaceutical composition is a single-dose or multi-dose aqueous injectable pharmaceutical composition.

[0032] Examples of one or more GLP-1 receptor agonists include, but are not limited to, GLP-1, GLP-1 analogs, and derivatives of GLP-1 analogs.

[0033] In some embodiments, the stable pharmaceutical composition further comprises a second active ingredient, which comprises insulin, an insulin analog, and / or a derivative of an insulin analog, and may be referred to as a stable combination pharmaceutical composition in this invention. Embodiments of the stable combination pharmaceutical compositions disclosed in this invention also exhibit high stability of the first and second active ingredients. In some embodiments, stability relates to physical stability. In some embodiments, the stable combination pharmaceutical composition is a stable liquid combination pharmaceutical composition. In some embodiments, the stable liquid combination pharmaceutical composition is a stable aqueous combination pharmaceutical composition. In some embodiments, the stable aqueous combination pharmaceutical composition is a stable aqueous injectable combination pharmaceutical composition. In some embodiments, the stable aqueous injectable combination pharmaceutical composition is a single-dose or multi-dose aqueous injectable combination pharmaceutical composition.

[0034] In some embodiments of the stable combination pharmaceutical compositions disclosed in this invention, the second active ingredient comprises insulin or an insulin analogue. In some embodiments of the stable combination pharmaceutical compositions disclosed in this invention, the insulin analogue includes 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 in this invention, based on the total weight of the stable combination pharmaceutical composition, the stable combination pharmaceutical composition comprises about 0.1% wt to about 1% wt of insulin, an insulin analogue, or a derivative of an insulin analogue, optionally at least about 0.1% wt to about 0.4% wt of insulin, an insulin analogue, or a derivative of an insulin analogue, for example, at least about 0.2% wt to about 0.4% wt of insulin or an insulin analogue. For example, the stable combination drug composition may contain at least one of insulin lispro, insulin aspart, insulin glutares, insulin glargine, insulin degludec, and insulin detemir, having a concentration of 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 comprises insulin, an insulin analog, or a derivative of an insulin analog having a concentration of at least about 0.20% wt to about 0.40% wt, such as, but not limited to, about 0.21% wt, about 0.22% wt, about 0.23% wt, about 0.24% wt, about 0.25% wt, about 0.26% wt, about 0.27% wt, about 0.28% wt, about 0.29% wt, about 0.30% wt, about 0.31% wt, about 0.32% wt, about 0.33% wt, about 0.34% wt, about 0.35% wt, about 0.36% wt, about 0.37% wt, about 0.38% wt, about 0.39% wt, or about 0.40% wt. It should be noted that when the stable combination pharmaceutical composition is formulated as a liquid pharmaceutical composition, such as a premixed injectable water composition, the aforementioned "%wt" may also be expressed as "mg / mL". For example, 0.40% by weight of insulin or an insulin analog or derivative of insulin analog also means 4.0 mg / mL of insulin or an insulin analog or derivative of insulin analog. As shown in the examples provided in the Examples section, alkyl glycosides (e.g., but not limited to DDM) unexpectedly improved the stability of GLP-1 receptor agonists, and also simultaneously improved the stability of GLP-1 receptor agonists and insulin or derivatives of insulin analog.In some embodiments, the first active ingredient comprises or is benaglutide, and the second active ingredient comprises or is insulin glargine. In some embodiments, the first active ingredient comprises or is benaglutide, and the second active ingredient comprises or is insulin aspart.

[0035] In certain embodiments of the stable pharmaceutical compositions disclosed in this invention, examples of alkyl glycosides comprise a glycosyl group and an alkyl group linked by a linking group. The glycosyl 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 comprises about 10 to about 14 carbon atoms. In some embodiments, the alkyl group comprises about 6 to about 20 carbon atoms. In some embodiments, the alkyl group comprises 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 comprises unbranched alkyl groups and / or branched alkyl groups. In some embodiments, the alkyl group comprises lower alkyl groups (e.g., C1-C8) substituted with cycloalkyl groups. Examples of lower alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, hexyl, isohexyl, heptyl, isohexyl, octyl, and isooctyl. Examples of cycloalkyl groups include, but are not limited to, cyclohexyl. In some embodiments, alkyl glycosides (e.g., but not limited to DDM) improve the stability of the stable pharmaceutical compositions disclosed herein. In some embodiments, the glycoside comprises maltose or sucrose. In some embodiments, the alkyl glycoside comprises a group selected from n-dodecyl β-D-maltose glycoside (DDM), sucrose dodecanoate, n-dodecyl β-D-maltose glycoside, 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- At least one compound selected from β-D-glucopyranoside, n-octyl-β-D-maltose, n-nonyl-β-D-maltose, n-decyl-β-D-maltose, cyclohexyl-methyl-β-D-maltose, 2-cyclohexyl-ethyl-β-D-maltose, 3-cyclohexyl-propyl-β-D-maltose, 4-cyclohexyl-butyl-β-D-maltose, or 5-cyclohexyl-pentyl-β-D-maltose.

[0036] In some embodiments, based on the total weight of the stable pharmaceutical composition, the stable pharmaceutical composition comprises about 0.05% wt to about 0.50% wt of an alkyl glycoside. For example, the concentration of the alkyl glycoside is at least about 0.05% wt, about 0.06% wt, about 0.07% wt, about 0.08% wt, about 0.09% wt, about 0.10% wt, about 0.11% wt, about 0.12% wt, about 0.13% wt, about 0.14% wt, about 0.15% wt, about 0.16% wt, about 0.17% wt, about 0.18% wt, about 0.19% wt, about 0.20% wt, about 0.21% wt, about 0.22% wt, about 0.23% wt, about 0.24% wt, or about 0.24% wt. The concentrations are approximately 0.25%wt, 0.26%wt, 0.27%wt, 0.28%wt, 0.29%wt, 0.30%wt, 0.31%wt, 0.32%wt, 0.33%wt, 0.34%wt, 0.35%wt, 0.36%wt, 0.37%wt, 0.38%wt, 0.39%wt, 0.40%wt, 0.41%wt, 0.42%wt, 0.43%wt, 0.44wt, 0.45%wt, 0.46%wt, 0.47%wt, 0.48%wt, 0.49%wt, or about 50%wt. In some embodiments, the concentration of the alkyl glycoside is at least about 0.10%wt to about 0.20%wt. In some embodiments, the concentration of the alkyl glycoside is at least about 0.10% wt to about 0.30% wt.

[0037] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the second active ingredient comprises or is insulin aspart, the surfactant is one or more alkyl glycosides (e.g., but not limited to DDM), and the concentration of said surfactant is about 0.05%wt to about 0.50%wt, about 0.05%wt to about 0.45%wt, about 0.05%wt to about 0.40%wt, about 0.05%wt to about 0.35%wt, about 0.05%wt to about 0.30%wt, about 0.05%wt to about 0.25%wt, 0.05%wt to about 0.20%wt, 0 0.05%wt to about 0.15%wt, 0.05%wt to about 0.10%wt, 0.10%wt to about 0.50%wt, 0.10%wt to about 0.45%wt, about 0.10%wt to about 0.40%wt, about 0.10%wt to about 0.35%wt, about 0.10%wt to about 0.30%wt, about 0.10%wt to about 0.25%wt, about 0.10%wt t to about 0.20%wt, about 0.10%wt to about 0.15%wt, about 0.16%wt to about 0.30%wt, about 0.16%wt to about 0.25%wt, about 0.20%wt to about 0.30%wt, about 0.20%wt to about 0.25%wt, about 0.05%wt, about 0.10%wt, about 0.16%wt, about 0.20%wt, or about 0.25%wt.

[0038] In some embodiments, the GLP-1 receptor agonist is a peptide agonist. Optionally, the GLP-1 receptor agonist can also act as a GIP agonist. In other words, dual agonists (e.g., but not limited to P13) can also be used.

[0039] In some embodiments, the GLP-1 receptor agonist comprises at least one selected from: GLP-1, GLP-1 analogs or derivatives thereof; and truncated GLP-1 fragments or analogs or derivatives thereof. Examples of GLP-1 analogs include, but are not limited to, 988927, exendin-4, and their analogs and derivatives. In some embodiments, the GLP-1 receptor agonist comprises at least one selected from: liraglutide, exenatide, lixisenatide, albiglutide, benaglutide, dulaglutide, semaglutide, pepamotidide, and tilpotide. In some embodiments, the GLP-1 receptor agonist includes or is benaglutide.

[0040] As used in this invention, "GLP-1 receptor agonist" refers to a polypeptide comprising a natural, elongated, or truncated GLP-1 polypeptide (e.g., but not limited to GLP-1(7-37)OH / NH2, GLP-1(7-36)OH / NH2, GLP-1(7-35)OH / NH2), a GLP-1 fragment, a GLP-1 analog, 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., but not limited to P13). As used in this invention, a GLP-1 compound or a GLP-1 receptor agonist can bind to the GLP-1 receptor and initiate a signal transduction pathway leading to insulinotropic activity. GLP-1 and GLP-1 analogs may have a C-terminal free carboxyl group or a C-terminal amide group. For example, a GLP-1 analog can be a recombinant human GLP-1 (7-36) peptide with 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, called benaglutide. The molecular formula of benaglutide is C7. 149 H 225 N 39 O 46 It has a molecular weight of 3298.7. Except for endogenous amidation, where the native form of NH2 is replaced by an OH group in the recombinant peptide, benaglutide is essentially identical to the active form of GLP-1 in vivo. Benaglutide has a free carboxyl group at the C-terminus.

[0041] In certain embodiments of the stable pharmaceutical compositions disclosed in this invention, the GLP-1 receptor agonist is a polypeptide having the following nucleotide sequence: XHXEGTXTSDXSXXXEXXAXXXFIXWLXXGXX (SEQ ID NO: 1), wherein, A single X is: R, or missing; The 3-digit X can be A, G, V, L, I, S, or T; The 4-digit X is: F, W, or Y; The 11-bit X is: V, S, W, I, L, K, F, or Y; The 13-bit X can be: S, W, Y, F, K, I, L, or V; The 14-bit X can be: Y, W, or F; The 15-bit X can be: L, F, Y, or W; The 17-bit X can be G, E, D, Q, N, K, R, or C; The 18-bit X is: H, D, K, E, or Q; The 20-bit X can be A, V, I, or L; The 21-bit X is: K, R, Q, or N; The 22-bit X can be A, E, H, F, Y, W, R, I, or K; The 25-bit X can be A, E, D, S, or H; The 28-bit X is either V or I; The 29-bit X can be: K, R, Q, or N; The 31-bit X is: R, R-NH2, K, or K-NH2; The 32-bit X is: G, K, R, T, S, E, D, W, Y, F, H or missing.

[0042] In certain embodiments of the stable pharmaceutical compositions disclosed in this invention, the GLP-1 receptor agonist is a polypeptide having the following sequence: XXXGTXXXXXSKQXEEEAVXLXXXXLXXLKNGGXXXXXXXXXX (SEQ ID NO: 2), wherein: The 1-bit X is: H, R, Y, or EL; The 2-digit X is: S, G, A, or T; The 3-digit X is either D or E; The 6-digit X is either F or Y; The 7-bit X can be T, Y, or S; The 8-bit X is either S or Y; The 9-bit X is either D or E; The 10-bit X is either L or I; The 14-bit X is: L, I, V, or M; The 20-bit X is either R or K; The 22-bit X is either F or Y; The 23-bit X is: I, V, L, or M; The 24-bit X is either E or D; The 25-bit X is: W, F, or Y; The 31-bit X is either P or missing; The 32-bit X is: S or missing; The 33-bit X is either S or missing; The 34-bit X is: G or missing; The 35-bit X is either A or missing; A 36-bit X is either P or missing; The 37-bit X is either P or missing; The 38-bit X is either P or missing; The 39-bit X is: S, R, or missing; and The X at position 40 can be any amino acid or be omitted.

[0043] Typically, the stability of GLP-1 receptor agonists can be compromised by a variety of stresses. These stresses include pH, temperature, light, vibration, and the gas-liquid interface. A range of commonly used surfactants have also been tested for their potential to improve 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 tested but not effective in improving stability include, for example, polysorbate 20, polysorbate 80, poloxamer 188, hydroxypropyl β-cyclodextrin, and amino acids. The present invention also provides embodiments of stable pharmaceutical compositions that do not contain any surfactant selected from polysorbate 20, polysorbate 80, poloxamer 188, and hydroxypropyl β-cyclodextrin. In some embodiments, the stable pharmaceutical compositions do 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., but not limited to DDM).

[0044] In some embodiments, the stable pharmaceutical composition is a liquid formulation. In some embodiments, the stable pharmaceutical composition is a premixed aqueous injectable formulation. In some embodiments, the stable pharmaceutical composition is formulated as a premixed injectable composition, preferably for subcutaneous injection.

[0045] In some embodiments of the stable pharmaceutical compositions disclosed in this invention, the pharmaceutical compositions optionally include other pharmaceutically acceptable excipients, such as, but not limited to, buffer systems, antibacterial agents, pharmaceutically acceptable solubilizers, and zinc ions.

[0046] In some embodiments, the stable pharmaceutical compositions disclosed in this invention have a pH of 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 stable pharmaceutical compositions disclosed in this invention have a pH of about 3.0, 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, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, or about 5... .7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, about 8.5, about 8.6, about 8.7, about 8.8, about 8.9, or about 9.0.

[0047] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs) and has a pH range of 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.

[0048] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the pH is about 6.5 to about 8.5, about 6.6 to about 8.4, about 6.7 to about 8.3, about 6.8 to about 8.2, about 6.9 to about 8.2, about 7.2 to about 8.2, about 7.0 to about 8.15, about 7.0 to about 8.1, about 7.0 to about 8.0, about 7.0 to about 7.9, about 7.0 to about 7.8, about 7.0 to about 7.7, about 7.0 to about 7.6, or about 7. 1 to 8.2, about 7.1 to 8.15, about 7.1 to 8.1, about 7.1 to 8.0, about 7.1 to 7.9, about 7.1 to 7.8, about 7.1 to 7.7, about 7.1 to 7.6, about 7.2 to 8.2, about 7.2 to 8.15, about 7.2 to 8.1, about 7.2 to 8.0, about 7.2 to 7.9, about 7.2 to 7.8, about 7.2 to 7.7, about 7.2 to 7.6, about 7.3 to 8.2, about 7.3 to 8.15, about 7.3 to 8.1, about 7.3 to 8.0, about 7.3 to 7.9, about 7.3 to 7.8, about 7.3 to 7.7, about 7.3 to 7.6.

[0049] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the second active ingredient comprises or is insulin glargine, and the pH is about 3.5 to about 4.6, about 3.5 to about 4.5, about 3.5 to about 4.4, about 3.5 to about 4.3, about 3.5 to about 4.2, about 3.5 to about 4.1, about 3.5 to about 4.0, 3.6 to about 4.6, about 3.6 to about 4.5, about 3.6 to about 4.4, about 3.6 to about 4.3, or about 3.6 to about 4.2. Approximately 3.6 to approximately 4.1, approximately 3.6 to approximately 4.0, approximately 3.7 to approximately 4.6, approximately 3.7 to approximately 4.5, approximately 3.7 to approximately 4.4, approximately 3.7 to approximately 4.4, approximately 3.7 to approximately 4.3, approximately 3.7 to approximately 4.2, approximately 3.7 to approximately 4.1, approximately 3.7 to approximately 4.0, approximately 3.8 to approximately 4.6, approximately 3.8 to approximately 4.5, approximately 3.8 to approximately 4.4, approximately 3.8 to approximately 4.3, approximately 3.8 to approximately 4.2, approximately 3.8 to approximately 4.1, approximately 3.8 to approximately 4.0, approximately 3.9 to approximately 4.6, approximately 3.9 to approximately 4.5, approximately 3.9 to approximately 4.4, approximately 3.9 to approximately 4.3, approximately 3.9 to approximately 4.2, approximately 3.9 to approximately 4.1, approximately 3.9 to approximately 4.0.

[0050] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the second active ingredient comprises or is insulin aspart, and the pH is about 6.5 to about 8.5, about 6.6 to about 8.4, about 6.7 to about 8.3, about 6.8 to about 8.2, about 6.9 to about 8.2, about 7.0 to about 8.2, about 7.0 to about 8.15, about 7.0 to about 8.1, about 7.0 to about 8.0, about 7.0 to about 7.9, about 7.0 to about 7.8, about 7.0 to about 7.7, about 7.0 to about 8.5, about 6.6 to about 8.4, about 6.7 to about 8.3, about 6.8 to about 8.9, about 7.0 to about 7.8, about 7.0 to about 7.7, about 7.0 to about 8.9, about 7.0 to about 7.8, about 7.0 to about 7.7, about 7.0 to about 8.9, about 7.0 to about 7.8, about 7.0 to about 7.7, about 7.0 to about 8.9, about 7.0 to about 7.8, about 7.0 to about 7.9 ... About 7.6, about 7.1 to about 8.2, about 7.1 to about 8.15, about 7.1 to about 8.1, about 7.1 to about 8.0, about 7.1 to about 7.9, about 7.1 to about 7.8, about 7.1 to about 7.7, about 7.1 to about 7.6, about 7.2 to about 8.2, about 7.2 to about 8.15, about 7.2 to about 8.1, about 7.2 to about 8.0, about 7.2 to about 7.9, about 7.2 to about 7.8, about 7.2 to about 7.7, about 7.2 to about 7.6, about 7.3 to about 8.2, about 7.3 to about 8.15, about 7.3 to about 8.1, about 7.3 to about 8.0, about 7.3 to about 7.9, about 7.3 to about 7.8, about 7.3 to about 7.7, or about 7.3 to about 7.6.

[0051] In some embodiments, the stable pharmaceutical composition comprises one or more buffer systems commonly used for pH adjustment. Such buffer systems may comprise one or more buffer salts. Examples of 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 (e.g., but not limited to acetates, chlorides, and phosphates) may be from about 0 mM to about 50 mM, from about 0 mM to about 45 mM, from about 0 mM to about 40 mM, from about 0 mM to about 35 mM, from about 0 mM to about 30 mM, from about 0 mM to about 25 mM, from about 0 mM to about 20 mM, from about 10 mM to about 50 mM, from about 10 mM to about 45 mM, from about 10 mM to about 40 mM, from about 10 mM to about 35 mM, from about 10 mM to about 30 mM, from about 10 mM to about 25 mM, and from about 10 mM to about 20 mM. In some embodiments, the stable pharmaceutical composition may not contain a buffer salt.

[0052] In some embodiments, the stable pharmaceutical composition comprises zinc ions. In some embodiments, the concentration range of zinc ions is from about 1 μg / mL to about 100 μg / mL, from about 10 μg / mL to about 70 μg / mL, from about 10 μg / mL to about 50 μg / mL, from about 13 μg / mL to about 33 μg / mL, from about 20 μg / mL to about 40 μg / mL, from about 25 μg / mL to about 40 μg / mL, from about 10 μg / mL, from about 15 μg / mL, from about 20 μg / mL, from about 25 μg / mL, from about 30 μg / mL, from about 35 μg / mL, from about 40 μg / mL, from about 45 μg / mL, or from about 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 from about 1:4 to about 10.8:6.

[0053] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the second active ingredient comprises or is insulin glargine, and the stable combination pharmaceutical composition further comprises, at concentrations of about 1 μg / mL to about 100 μg / mL, about 10 μg / mL... Zinc ions at concentrations of approximately 70 μg / mL, approximately 10 μg / mL, approximately 50 μg / mL, approximately 13 μg / mL, approximately 33 μg / mL, approximately 20 μg / mL, approximately 40 μg / mL, approximately 25 μg / mL, 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 the 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.

[0054] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1, such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the second active ingredient comprises or is insulin aspart, and the stable combination pharmaceutical composition further comprises at concentrations of about 1 μg / mL to about 100 μg / mL, about 10 μg / mL... Zinc ions at concentrations of 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 the 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.

[0055] In some embodiments, the stable pharmaceutical composition may comprise one or more types of anions. Examples of anions include, but are not limited to, chloride ions, thiocyanate ions (KSCN), p-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.

[0056] In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition as disclosed in this invention, wherein the first active ingredient comprises one or more GLP-1 receptor agonists (e.g., GLP-1 such as benaglutide, GLP-1 analogs, and derivatives of GLP-1 analogs), and the second active ingredient comprises or is insulin aspart or insulin glargine, and the stable combination pharmaceutical composition further comprises anion, such as, but not limited to, Cl. - KSCN - PABA - and 4H3N - In some embodiments, the concentration of anions (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.

[0057] In some embodiments, the stable pharmaceutical composition comprises one or more solubilizers. As used herein, the term "solvent" refers to a solvent that contributes to or enhances solubility, thereby improving solubility. The term "solvent" as used herein does not include surfactants. In some embodiments, examples of solubilizers are, but are not limited to, glycerol, mannitol, propylene glycol, or combinations thereof. Based on the total volume of the stable pharmaceutical composition, the solubilizer concentration can be from about 0 mg / mL to about 40 mg / mL, from about 0 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 15 mg / mL, from about 20 mg / mL, from about 25 mg / mL, from about 30 mg / mL, from about 35 mg / mL, from about 40 mg / mL, or from about 45 mg / mL. In some embodiments, the solubilizer is glycerol. Based on the total volume of the stable pharmaceutical composition, in some embodiments, the solubilizer is glycerol or propylene glycol having a concentration of about 0 mg / mL to about 40 mg / mL, about 10 mg / mL to about 25 mg / mL, about 12 mg / mL, about 14 mg / mL, about 16 mg / mL, about 17 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, or about 24 mg / mL. Based on the total volume of the stable pharmaceutical composition, in some embodiments, the solubilizer is mannitol with a concentration of about 0 mg / mL to about 50 mg / mL, 0 mg / mL to about 40 mg / mL, about 10 mg / mL to about 25 mg / mL, about 12 mg / mL, about 14 mg / mL, about 16 mg / mL, about 17 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, or about 24 mg / mL.

[0058] In some embodiments, the stable pharmaceutical composition comprises one or more osmotic pressure regulators, such as, but not limited to, glycerol, mannitol, propylene glycol, or combinations thereof.

[0059] In some embodiments, the stable pharmaceutical composition is an isotonic system of the subject's plasma, typically having an osmotic pressure of about 280 mOsm / L to about 320 mOsm / L. In some embodiments, the stable pharmaceutical composition has an osmotic pressure of about 280 mOsm / L to about 320 mOsm / L, about 270 mOsm / L to about 330 mOsm / L, or about 260 mOsm / L to about 340 mOsm / L. In some embodiments, the pharmaceutical composition comprises 40 mg / mL mannitol and 5 mg / mL propylene glycol as osmolarity regulators. In some embodiments, the stable pharmaceutical composition is an insulin glargine injection composition or an insulin glargine / GLP-1 combination pharmaceutical composition, wherein the osmolarity regulator is glycerol. The glycerol concentration in the insulin glargine injection composition is 17 mg / mL, and the glycerol concentration in the combination pharmaceutical composition is 20 mg / mL. Other suitable osmolarity regulators include, but are not limited to, mannitol, glycerol, propylene glycol, and combinations thereof.

[0060] In some embodiments, the stable pharmaceutical composition comprises one or more antibacterial agents. Optionally, the antibacterial agent comprises at least one compound selected from phenolic compounds, aromatic alcohols, phenoxyethanol, and chloroethanol compounds. In some embodiments, the antibacterial agent comprises at least one compound selected from phenol, m-cresol, benzyl alcohol, phenoxyethanol, or chlorobutanol. In some embodiments, the antibacterial agent is phenol. In some embodiments, the stable pharmaceutical composition comprises concentrations of about 2.0 mg / mL to about 4.0 mg / mL, about 2.0 mg / mL, about 2.1 mg / mL, about 2.2 mg / mL, about 2.3 mg / mL, about 2.4 mg / mL, about 2.5 mg / mL, about 2.6 mg / mL, about 2.7 mg / mL, about 2.8 mg / mL, about 2.9 mg / mL, about 3.0 mg / mL, about 3.1 mg / mL, about 3.2 mg / mL, about 3.3 mg / mL, about 3.4 mg / mL, about 3.5 mg / mL, about 3.6 mg / mL, about 3.7 mg / mL, about 3.8 mg / mL, about 3.9 mg / mL, about 4.0 mg / mL, about 2.10 mg / mL, about 2.11 mg / mL, about 2.12 mg / mL, and about 2.13 mg / mL. Approximately 2.14 mg / mL, approximately 2.15 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 Phenol at concentrations of 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.

[0061] In some embodiments, the stable pharmaceutical composition may comprise concentrations of about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 2.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1.0 mg / mL, about 1.1 mg / mL, about 1.2 mg / mL, about 1.3 mg / mL, about 1.4 mg / mL, about 1.5 mg / mL, about 1.6 mg / mL, about 1.7 mg / mL, about 1.8 mg / mL, and about 1 mg / mL. GLP-1 receptor agonists at concentrations of approximately 2.0 mg / mL, 2.1 mg / mL, 2.2 mg / mL, 2.3 mg / mL, 2.4 mg / mL, 2.5 mg / mL, 2.6 mg / mL, 2.7 mg / mL, 2.8 mg / mL, 2.9 mg / mL, 3.0 mg / mL, 3.1 mg / mL, 3.2 mg / mL, 3.3 mg / mL, 3.4 mg / mL, 3.5 mg / mL, 3.6 mg / mL, 3.7 mg / mL, 3.8 mg / mL, 3.9 mg / mL, or 4.0 mg / mL.

[0062] In some embodiments of the stable pharmaceutical compositions disclosed in this invention, no foreign matter formation was observed in an accelerated stability test at 25°C for 3 months (25C3M), and the peptide (i.e., the amount of peptide before the test exceeds the amount of peptide after the test) was not less than about 93%, or not less than about 94%. Examples of the corresponding methods are described in the General Methods section of the Examples section.

[0063] Another aspect of the present invention relates to a method for improving the storage stability of a pharmaceutical composition comprising a first active ingredient containing one or more GLP-1 receptor agonists. In some embodiments of the method disclosed herein, the method includes mixing one or more surfactants (e.g., but not limited to, alkyl glycosides (e.g., but not limited to DDM)) 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 glutathione, insulin glargine, insulin degludec, and insulin detemir.

[0064] In some embodiments, one or more alkyl glycosides include at least one selected from DDM, sucrose dodecanoate, n-dodecyl β-D-maltodextrin, 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-maltodextrin, n-nonyl-β-D-maltodextrin, n-decyl-β-D-maltodextrin, cyclohexyl-methyl-β-D-maltodextrin, 2-cyclohexyl-ethyl-β-D-maltodextrin, 3-cyclohexyl-propyl-β-D-maltodextrin, 4-cyclohexyl-butyl-β-D-maltodextrin, or 5-cyclohexyl-pentyl-β-D-maltodextrin. In some embodiments, the alkyl glycoside is DDM.

[0065] Another aspect of the invention relates to the use of the stable pharmaceutical composition in the preparation of a medicament for: (i) preventing and / or treating diabetes, and / or lowering HbA1C; (ii) delaying or preventing the progression of diabetes, delaying the progression of impaired glucose tolerance to insulin-dependent type 2 diabetes, delaying or preventing insulin resistance and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes; (iii) improving β-cell function, and / or restoring β-cell glucose sensitivity; (iv) preventing and / or treating eating disorders; decreased gastric motility; delayed gastric emptying; (v) preventing and / or treating complications of obesity; (vi) preventing and / or treating complications of diabetes; and / or (vii) preventing and / or treating cardiovascular disease. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog.

[0066] Another aspect of the invention relates to the use of stable pharmaceutical compositions for: (i) prevention and / or treatment of diabetes, and / or reduction of HbA1C; (ii) delaying or preventing the progression of diabetes, delaying the progression of impaired glucose tolerance to insulin-dependent type 2 diabetes, delaying or preventing insulin resistance and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes; (iii) improvement of β-cell function, and / or restoration of β-cell glucose sensitivity; (iv) prevention and / or treatment of eating disorders; decreased gastric motility; delayed 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. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog.

[0067] Another aspect of the invention relates to methods for: (i) preventing and / or treating diabetes, and / or lowering HbA1C; (ii) delaying or preventing the progression of diabetes, delaying the progression of impaired glucose tolerance to insulin-dependent type 2 diabetes, delaying or preventing insulin resistance and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes; (iii) improving β-cell function, and / or restoring β-cell glucose sensitivity; (iv) preventing and / or treating eating disorders; decreased gastric motility; delayed gastric emptying; (v) preventing and / or treating complications of obesity; (vi) preventing and / or treating complications of diabetes; and / or (vii) preventing and / or treating cardiovascular disease, comprising administering a therapeutically effective amount of a stable pharmaceutical composition to a subject. In some embodiments, the stable pharmaceutical composition is a stable combination pharmaceutical composition further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of an insulin analog.

[0068] The following embodiments are provided to better illustrate the claimed invention and the embodiments described herein, but these embodiments should not be construed as limiting the scope of the invention. Unless otherwise stated, the specific materials listed in the following embodiments are for illustrative purposes only and are not intended to limit the invention. Various equivalents, changes, and modifications can be made by those skilled in the art without departing from the scope of the invention, and it should be understood that these equivalent embodiments are intended to be included within the scope of the invention. Furthermore, all references cited in this invention are incorporated herein by reference in their entirety as if fully set forth herein.

[0069] Example General methods Unless otherwise stated, the methods described in the General Methods section are used in the embodiments described in the Examples section.

[0070] I. ThT test The ThT assay is a commonly used method for monitoring protein fibrillation. Thionyl T (ThT) is a benzothiazole fluorescent dye that binds highly specifically to amyloid fibrils and causes fluorescence enhancement upon binding. This assay uses changes in fluorescence value to measure the kinetics of protein fibrillation. Fibrillation occurs when the fluorescence value exceeds a predetermined threshold. Unless otherwise stated, in embodiments of the invention, the predetermined threshold is three times the initial fluorescence value at the measurement point, i.e., Tn / T1 = 3.

[0071] ThT test plan: 1. Sample preparation (according to the ThT test sample formulation table): Mix the test sample (500 μL) with thiosulfate T (aq, 1 mM, 25 μL) to provide a first mixture. Add 200 μL of the first mixture to a black V-shaped 96-well plate (Eppendorf) and set up two replicate wells for each sample (each sample is applied to two wells for measurement).

[0072] 2. Sample incubation: Cover the 96-well plate with sealing tape and seal it to prevent sample evaporation and contamination; place the 96-well plate in a mixer (Thermo Mixer C, Eppendorf) and incubate in the dark at 37°C and 300 rpm. The 96-well plate used is a 96-well Costa blk / clrbtm (Costar laboratory 96-well plate with a black bottom and a clear top), although other suitable 96-well plates may also be used.

[0073] 3. Fluorescence measurement parameters: On the M5e multi-functional microplate reader (Molecular Devices, LLC), the fluorescence measurement mode was set with an excitation wavelength of 430 nm, an emission wavelength of 470 nm, a measurement temperature of room temperature (around 25℃), and a 96-well plate of 96-well Costa blk / clrbtm.

[0074] 4. Sample Measurement: Incubate the test samples for the set time to perform fluorescence measurement, and take the average of the fluorescence measurements from two replicate wells for data analysis. The initial fluorescence value is the fluorescence measurement value after incubation at 37°C for 1 hour. After removing the 96-well plate, wrap it with aluminum foil to protect it from light.

[0075] II. Stability Test Stability studies consist of two parts: visual inspection and peptide content testing. Visual inspection assesses stability by visually observing the transparency of the test composition and the formation of any visible foreign matter. Peptide content testing determines the amount of peptides in the composition using HPLC. Compositions that pass the visual inspection are selected and subjected to accelerated testing in a 25°C stability testing chamber, with peptide content testing performed at the designated time points.

[0076] 1. Visual inspection Samples were prepared according to the stability test formulation and stored in a stability testing chamber at 25°C for accelerated testing. Samples were removed and visually inspected at different time points (e.g., 14 days, 1 month, 2 months, 3 months, etc.). Visual inspection involved observing indicators such as the composition's transparency and the formation of visible foreign matter. If no obvious abnormalities were observed during visual inspection, the composition met the requirements and was proceeded to the next step of HPLC testing. Otherwise, the composition did not meet the requirements, and no further measurements were performed.

[0077] 2. Detection of peptide content The compositions used for stability testing were stored in a stability testing chamber at 25°C to accelerate the testing. The peptide content of the compositions that passed visual inspection was examined at different time points (e.g., 14 days, 1 month, 2 months, 3 months, etc.). 100 μL of the composition was taken as a test sample. The amounts of GLP-1 and insulin glargine in the composition were measured using HPLC to evaluate stability.

[0078] The chromatographic 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 (pH 2.5)-sodium chloride-25% acetonitrile, and mobile phase B was phosphate (pH 2.5)-sodium chloride-65% acetonitrile. A gradient elution program was used, with the concentration of phase B gradually increasing from 4% to 17% within 20 minutes, and then further increasing to 53% within 18 minutes for gradient elution.

[0079] Under the same chromatographic conditions, chromatograms of glargine insulin and GLP-1 as individual components can be obtained.

[0080] The peaks of glargine insulin and GLP-1 or their derivatives can be identified by comparing the chromatogram of the combined composition with the chromatogram of the single-component composition of glargine insulin and GLP-1, respectively. Generally, glargine insulin and / or its derivatives have relatively weak retention on a C18 column, and their peak positions are in the first half of the chromatogram of the combined drug composition. GLP-1 and / or its derivative, benaglutide, have relatively strong retention, and their peak positions are in the second half of the chromatogram of the drug composition. The percentage of remaining GLP-1 peptide (also referred to as "GLP-1 peptide %" or "peptide %" unless otherwise stated) can be obtained by calculating the ratio of the peak area of ​​GLP-1 at each stability detection time point to the initial value. In other words, at different stability detection time points, the composition sample is taken for HPLC analysis to obtain the peak area corresponding to the chromatographic peak of GLP-1, and then the GLP-1 peak area at each stability detection time point is compared with the GLP-1 peak area at the initial time point (t0). The glargine insulin peptide % can be calculated using the same method.

[0081] Example 1 A series of pharmaceutical compositions (compositions 1 to 28) containing GLP-1 receptor agonists were prepared using the various components listed in Table 1, with zinc acetate used as the zinc ion source. ThT tests were performed on these compositions, showing that DDM significantly inhibited the formation of GLP-1 fibrosis and improved the physical stability of the pharmaceutical compositions. Results are referenced separately. Figures 1 to 8 discuss.

[0082] Table 1 Compositions 1 to 28

[0083] Summary of ThT test results for compositions 1-28 1. ThT test results of compositions 1 to 4 ( Figure 1 The results showed that for the glargine insulin / benalutide stock solution combination test samples, DDM inhibited the formation of GLP-1 fibrinolysis and exhibited significantly better physical stability than other surfactants tested (e.g., Tween 20 and Tween 80). Figure 1As shown, the increase in fluorescence was not significant before 30 hours (early stage), therefore, no obvious trend of fibril formation was observed. When the incubation time was extended beyond 30 hours, significant increases in fluorescence values ​​were observed in composition 1 (solid triangle, B+G_phenol) without surfactant, and in compositions 2 and 3 containing Tween 20 (hollow circle, B+G_phenol_0.05% TW20) and Tween 80 (solid square, B+G_phenol_0.05% TW80), respectively. However, no significant increase in fluorescence values ​​was observed in composition 4 containing DDM (hollow rhombus, B+G_phenol_0.05% DDM). The fluorescence values ​​of compositions 1-3 continued to increase rapidly over time, indicating that significant protein fibril formation occurred therein. However, in the test samples containing DDM, the fluorescence values ​​did not change significantly near 50 hours. Therefore, in the glargine insulin / benalutide stock solution combination test samples, DDM showed significantly better performance than other tested surfactants in inhibiting GLP-1 fibrillation and improving physical stability.

[0084] 2. ThT test results of compositions 5-8 ( Figure 2 The results showed that for benaglutide stock solution test samples, DDM significantly inhibited GLP-1 fibrillation formation and improved physical stability compared to other surfactants (e.g., Tween 20 and Tween 80). Figure 2 As shown, the increase in fluorescence was not significant before 30 hours (early stage), and therefore no obvious trend of fibril formation was observed. When the incubation time was extended beyond 30 hours, significant increases in fluorescence values ​​were observed in composition 5 (solid triangle, GLP-1_phenol) without surfactant, and in compositions 6 and 7 containing Tween 20 (hollow circle, GLP-1_phenol_0.05%TW20) and Tween 80 (solid square, GLP-1_phenol_0.05%TW80), respectively. However, no significant increase in fluorescence values ​​was observed in composition 8 containing DDM (hollow rhombus, GLP-1_phenol_0.05%DDM). The fluorescence values ​​of compositions 5-7 increased rapidly and continuously over time, indicating that significant protein fibril formation occurred therein. However, in the test samples containing DDM, the fluorescence values ​​did not change significantly near 50 hours. Therefore, in the benaglutide stock solution test samples, DDM showed significantly better performance than the other surfactants tested in inhibiting GLP-1 fibril formation and improving physical stability.

[0085] 3. ThT test results of compositions 9-12 ( Figure 3 The results showed that for the benaglutide composition test samples, DDM significantly inhibited the formation of GLP-1 fibrillation and improved physical stability. Figure 3As shown, the increase in fluorescence was not significant before 30 hours (early stage), and therefore no obvious trend of fibril formation was observed. When the incubation time was extended beyond 30 hours, a significant increase in fluorescence was observed in the surfactant-free composition 9 (solid square, BNLT_Lot1 (new)), while in the compositions containing DDM, even for compositions 10 (hollow circle, BNLT_Lot2+0.01%DDM) with a DDM concentration as low as 0.01%, and also for compositions 11 (solid triangle, BNLT_Lot2+0.05%DDM) with a DDM concentration of 0.05% and compositions 12 (hollow rhombus, BNLT_Lot2+0.1%DDM) with a DDM concentration of 0.1%. In the test samples containing DDM, the fluorescence value did not change significantly at nearly 50 hours. Therefore, in the benaglutide composition test samples, DDM showed a significant inhibition of GLP-1 fibril formation and improved physical stability.

[0086] 4. ThT test results of compositions 13-17 ( Figure 4 The results showed that, for compositions containing various antibacterial agents, DDM (0.05%) significantly inhibited the formation of protein fibrillation and improved physical stability. Figure 4 As shown, in the test samples of glargine insulin / benalutide stock solutions containing various antibacterial agents, the increase in fluorescence was not significant throughout the entire test time (>150 hours): composition 13 (solid square, B+G_phenol + 0.05% DDM) containing phenol, composition 14 (hollow circle, B+G_m-cresol + 0.05% DDM) containing m-cresol, composition 15 (solid triangle, B+G_benzyl alcohol + 0.05% DDM) containing benzyl alcohol, composition 16 (hollow rhombus, B+G_trichlorobutanol + 0.05% DDM) containing chlorobutanol, and composition 17 (square, B+G_phenoxyethanol + 0.05% DDM) containing phenoxyethanol. Therefore, in the compositions containing various antibacterial agents, DDM showed a significant inhibition of GLP-1 fibrosis formation and improved physical stability.

[0087] 5. ThT test results of compositions 18 to 22 ( Figure 5 Studies have shown that compositions containing DDM significantly inhibit protein fibrillation and improve physical stability compared to conventional compositions without DDM. Figure 5As shown, fluorescence was significantly increased in compositions 20 (solid square, 20201112-3), 21 (solid triangle, 20201112-4), and 22 (hollow circle, 20201112-5), which are conventional compositions without DDM. However, no significant fluorescence change was observed in compositions 18 (square, 20201112-1) and 19 (hollow rhombus, 20201112-2), which contain DDM. Therefore, it is shown that DDM is superior to conventional compositions without DDM in inhibiting protein fibrillation and physical stability.

[0088] 6. ThT test results of compositions 23 to 28 ( Figures 6-8 The results 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 inhibited the formation of GLP-1 fibrillation and improved physical stability. Figure 6 As shown, in composition 23 (without DDM) Figure 6 Hollow square, GLP 1-1 mg / mL), composition 25 ( Figure 7 Hollow square, P13) and composition 27 ( Figure 8 In the hollow square (988927) composition 24, fluorescence significantly increased after incubation at 37°C for 1-2 days. After 7 days of incubation, the fluorescence values ​​of compositions 23 and 27 increased by more than 100 times their initial values, and the fluorescence value of composition 25 increased by more than 35 times its initial value. However, no significant fluorescence change was observed in the compositions containing 0.01% DDM during the 1-week testing period: composition 24 ( Figure 6 Hollow triangle, GLP1-DDM), composition 26 ( Figure 7 Hollow triangle (P13-DDM), and composition 28 ( Figure 8 (Hollow triangle, 9889270DDM). Therefore, in compositions containing GLP-1, GLP-1 analogs (e.g., 988927) and GLP-1-based GLP-1 / GIP dual agonists (e.g., P13), DDM exhibits significant inhibition of GLP-1 fibrosis formation and improved physical stability.

[0089] 7. DDM improved the solubility of GLP-1 in the presence of phenol at neutral pH. GLP-1 solubility was evaluated relative to phenol concentration at neutral pH in the absence of DDM, with 0.2% DDM, and with 0.4% DDM. GLP-1 solubility decreased with increasing phenol concentration. Figure 9However, the presence of DDM improved GLP-1 solubility compared to the DDM-free GLP-1 composition (rhombus, DDM-free), and higher DDM concentrations (triangle, 0.4% DDM) showed further improvement in GLP-1 solubility compared to lower DDM concentrations (square, 0.2% DDM).

[0090] Example 2 A series of combination compositions containing GLP-1 receptor agonists and glargine insulin were prepared, and their stability tests showed that DDM improved the stability of the compositions.

[0091] Compositions A1 to A20 having the components listed in Table 2A were prepared, wherein zinc chloride was used as the zinc ion source. Visual inspection and peptide content testing were performed to determine stability.

[0092] Table 2A Compositions A1 to A20

[0093] Summary of stability test results 1. The stability test results of compositions A1–A4 are summarized in Table 2B. The data show that DDM significantly improves the stability of the compositions compared with other surfactants (e.g., Tween 20 and Tween 80).

[0094] Table 2B: Stability Test Results of Glargine Insulin / Benaglutide Stock Solution Composition

[0095] Y: Visual inspection passed.

[0096] *: Peptide% indicates the percentage of GLP-1 peptide in the composition after storage under specific conditions compared to the amount of GLP-1 before storage.

[0097] 1 :25C14D indicates that it can be stored at 25°C for 14 days.

[0098] 2 :25C1M indicates that it can be stored at 25°C for 1 month.

[0099] 3 :25C2M indicates that it can be stored at 25°C for 2 months.

[0100] 4 :25C3M indicates that it can be stored at 25°C for 3 months.

[0101] As shown in Table 2B, after one month of accelerated testing at 25°C (25°C 1M), the peptide percentage of the surfactant-free composition (composition A1, Lot20200519) decreased to 93.6%, while the peptide percentage of the composition containing Tween 20 (composition A2: Lot20200713 / BEM040+GLP1 1.0_TW20) decreased to 93.84%. After 14 days of accelerated testing at 25°C (25°C 14D), the composition containing Tween 80 (composition A3: Lot20200713 / BEM040+GLP1 1.0_TW80) showed gel formation and failed the appearance inspection. Composition A4 (composition A4: Lot20200928_4#), containing 0.1% DDM formulation, showed 94.66% peptide percentage after three months of accelerated testing at 25°C (25°C 3M) and passed the appearance inspection. Therefore, the experimental results show that DDM significantly increases the stability of the composition compared with Tween 20 and Tween 80.

[0102] 2. The stability test results of compositions A5–A9 are summarized in Table 2C. The data show that DDM significantly improves the stability of the compositions compared to conventional excipients.

[0103] Table 2C: Stability test results of the glargine insulin / benalutide stock solution composition

[0104] Y: Visual inspection passed.

[0105] *: Peptide% refers to the percentage of GLP-1 after storage under specific conditions relative to the amount of GLP-1 before storage.

[0106] As shown in Table 2C, formulations A7 to A9 use different combinations of glycerol, mannitol, and propylene glycol, but none contain DDM. Lowering the pH of the formulation helps improve physical stability and avoids appearance abnormalities. However, in the accelerated stability test (25°C 3M), when the pH was lowered, the peptide content of formulations A7 to A9 decreased to approximately 90% after 3 months. In contrast, the DDM-containing formulations (formulations A5-A6) maintained a peptide content of over 94% after three months of accelerated stability testing at 25°C. Therefore, compared to conventional excipients, DDM significantly improves the stability of the formulation.

[0107] 3. The stability test results of compositions A10–A14 are summarized in Table 2D. The data show that DDM significantly improved the stability of compositions containing various antibacterial agents.

[0108] Table 2D Stability test results of glargine insulin / benalutide stock solution-DDM compositions containing different antibacterial agents

[0109] Y: Visual inspection passed.

[0110] *: Peptide% refers to the percentage of GLP-1 after storage under specific conditions relative to the amount of GLP-1 before storage.

[0111] The antimicrobial agents tested are commonly used in injectable compositions, such as phenol, m-cresol, benzyl alcohol, chlorobutanol, and phenoxyethanol. All tested compositions contained DDM and passed an accelerated stability test at 25°C for 3 months, with a peptide content greater than 92%. Therefore, DDM not only significantly improves the stability of the compositions but is also compatible with a variety of antimicrobial agents commonly used in injection. 4. The stability test results of compositions A15–A18 are summarized in Table 2E. Compositions with different DDM concentrations are compared.

[0112] Table 2E Stability test results of glargine insulin / benalutide stock solution compositions containing different concentrations of DDM

[0113] *: Peptide% refers to the percentage of peptides stored under specific conditions relative to the amount of peptides before storage.

[0114] Composition A15 (0.05% DDM) failed visual inspection in an accelerated stability test at 25°C for 2 months (25C60D), while compositions containing 0.1% DDM or higher all passed visual inspection after an accelerated stability test at 25°C for 3 months (25C90D), with GLP-1 peptide % greater than 93%. Therefore, a DDM concentration of 0.1% or higher improves the physical stability of the composition to a greater extent than a DDM concentration of approximately 0.05%.

[0115] 5. The stability test results of compositions A16, A19, and A20 are summarized in Table 2E. Compositions containing DDM with different pH values ​​were compared.

[0116] Table 2F Results of stability study of glargine insulin / benalutide stock solution-DDM composition at different pH values

[0117] *: Peptide% refers to the percentage of peptides stored under specific conditions relative to the amount of peptides before storage.

[0118] As shown in Table 2F, within a pH range of 4.0–4.5, the combination composition containing 0.1% DDM showed no failures in the visual inspection during a three-month accelerated stability test at 25°C, and the peptide percentage was greater than 93%. Therefore, DDM effectively improves the stability of the formulation within the pH range of 4.0–4.5.

[0119] 6. A series of combination compositions (compositions A30-41) containing GLP-1 receptor agonists and insulin glargine were prepared, and their stability was tested.

[0120] Table 2G Composition A30-

[0121] As shown in Table 2H, after three months of accelerated stability testing at 25°C within a pH range of 3.8 to 4.4, the combination composition containing 0.1% DDM showed a glargine insulin peptide percentage greater than 97.23% and a GLP-1 peptide percentage greater than 95.49%. Therefore, DDM effectively improved the stability of the formulation within the pH range of 3.8 to 4.4.

[0122] Table 2H shows the results of stability studies on glargine insulin / benalutide-DDM combination compositions with different pH values.

[0123] As shown in Table 2I, after three months of accelerated stability testing at 25°C within a pH range of 3.8 to 4.4, the combination composition containing 0.1% DDM showed an insulin glargine peptide percentage greater than 97.23% and a GLP-1 peptide percentage greater than 95.49%. Therefore, DDM effectively improved the stability of the formulation within the pH range of 3.8–4.4.

[0124] Table 2.1 Stability study results of the glargine insulin / phenylglitazone-DDM combination with and without DDM.

[0125] Combinations containing different solubilizer combinations (glycerol / mannitol / propylene glycol) were tested with and without DDM. After accelerated testing at 40C28D, the test combination without DDM failed visual inspection. The presence of 0.1% DDM in the combination improved the appearance after accelerated testing at 40C28D, with GLP-1 peptide percentages exceeding 93% and 87% after accelerated testing at 40C14D and 40C28D, respectively.

[0126] Example 3 A series of combination compositions comprising a GLP-1 receptor agonist and insulin aspart were prepared, and stability tests showed that DDM improved the stability of the compositions. Unless otherwise stated, the buffer system for the combination composition in Example 3 was PB.

[0127] Compositions B1-B20 having the components listed in Table 3A were prepared. Visual inspection and peptide content testing were performed to determine stability.

[0128] Table 3A Compositions B1 to B20

[0129] 1. Compositions B1-B5: The combination drug compositions of the embodiments comprise insulin aspart and GLP-1 (benalutide) with different NaCl concentrations.

[0130] DLS (Dynamic Light Scattering) intensity spectroscopy is used to display the size of particles present in compositions B1-B5.

[0131] After 14 days of accelerated testing at 40°C (40C14D), the NaCl-free combination drug composition (composition B1) mostly had large GLP-1 particles above 100 nm, indicating that GLP-1 exhibits significant protein aggregation.

[0132] The content of large particles above 100 nm in the combined drug composition (composition B2) with a NaCl concentration of 10 mM is significantly reduced.

[0133] No large particles larger than 100 nm were observed in the combination drug compositions with NaCl concentrations of 20 mM (composition B3) or higher (compositions B4-B5). Most GLP-1 had a particle size of less than 10 nm.

[0134] The stability test results of compositions B1-B5 are summarized in Table 3B below.

[0135] Table 3B: Stability test results of the aspart insulin / GLP-1 combination composition

[0136] Following the 40C14D accelerated test, the conjugate composition without NaCl failed the visual inspection. When the NaCl concentration was increased to above 10 mM, the appearance of the conjugate composition significantly improved during the 40C14D accelerated test; thereafter, the percentage of insulin aspart peptide remained above 90.8%, and the percentage of GLP-1 peptide remained above 90.5%.

[0137] 2. Compositions B6-B8: Embodiments of combined pharmaceutical compositions of insulin aspart and GLP-1 (benalutide) with different DDM concentrations.

[0138] After 2 months of accelerated testing at 40°C (40°C 2M), composition B6 (0.16% DDM) failed the visual inspection, showing a white flocculent precipitate.

[0139] Composition B8 (0.25% DDM) failed the visual inspection after 14 days of accelerated testing at 40°C (40C14D), although the same composition passed the visual inspection after 3 months of storage at 4°C (4C3M).

[0140] Composition B7 (0.2% DDM) passed visual inspection after accelerated testing at 40°C for 14 days (40C14D) and after storage at 4°C for 3 months (4C3M).

[0141] 3. Compositions B9-B12: The combination drug compositions of the embodiments contain insulin aspart and GLP-1 (Benaglutide) with different pH values ​​(pH 7.4, 7.6 and 8.15).

[0142] The stability test results of compositions B1-B5 are summarized in Table 3B below.

[0143] Table 3B Stability test results of the aspart insulin / GLP-1 combination at different pH values

[0144] The combined composition at pH 8.15 (composition B9) maintained appearance stability for 3 months at 4°C; however, peptide % data showed that insulin aspart of composition B9 (pH 8.15) had a lower peptide content than that of composition B10 (pH 7.6) after the 40C2W accelerated assay.

[0145] Compositions B11 (pH 7.6) and B12 (pH 7.4) both passed visual inspection after the 40C 2W accelerated test, but both compositions maintained good flowability with foreign matter after the 40C 4W accelerated test.

[0146] 4. Compositions B13-B16: Embodiments of combination drug compositions (benalutide) of insulin aspart and GLP-1 with different zinc ion concentrations (1 μg / mL, 10 μg / mL, 20 μg / mL and 25 μg / mL).

[0147] The stability test results of compositions B1-B5 are summarized in Table 3C below.

[0148] Table 3C: Stability test results of the aspart insulin / GLP-1 combination at different zinc concentrations

[0149] Composition B13 contains EDTA, which can complex with all zinc in solution; therefore, composition B13 can be considered to be free of zinc ions. After storage at 4°C / 25°C / 40°C, the appearance and turbidity of the zinc-ion-free combined compositions (composition B13) and the combined compositions containing various concentrations of zinc ions (compositions B14-B16) were examined. After accelerated testing at 25°C / 40°C, the test compositions showed precipitation exceeding a predetermined value (>3.0), or after accelerated testing at 4°C, the test compositions also showed turbidity exceeding a predetermined value (>1.5).

[0150] Example 4 The pharmacokinetics (PK) of a combination drug composition (benalutide) of insulin aspart and GLP-1 was studied in beagle dogs (male, 2-3 years old, 9-11 kg).

[0151] After acclimatizing to the laboratory environment, 11 beagles were fasted for 20 hours without water, and then fed 4 hours after administration. The beagles were randomly assigned to three groups (n=3) based on their weight and were given GLP-1 alone (20 μg / kg, 50 μL / kg), insulin aspart alone (0.5 U / kg, 50 μL / kg), or a combination of GLP-1 (10 μg / kg) and insulin aspart (0.5 U / kg) (50 μL / kg).

[0152] Using a 1 mL syringe, flush with 1 mL of DPPIV inhibitor and protease inhibitor before and after administration. Collect 1.0 mL of whole blood rapidly from a beagle: for the GLP-1-only group, at 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, and 120 minutes after administration; for the aspartate insulin-only group and the combination group, at 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 120, 180, 240, 300, and 480 minutes after administration. Add the collected whole blood to an EDTA-K2 anticoagulant tube. Pre-add 10 μL of DPPIV inhibitor and 10 μL of protease inhibitor to the EDTA-K2 anticoagulant tube. Place the resulting mixture in an ice bath for 30 minutes before centrifugation (4°C, 300 g, 5 min) to collect plasma. 0.3 mL and 0.1 mL plasma samples were added to two separate test tubes, stored at -80°C, and used for the detection of active GLP-1 and insulin aspart, respectively.

[0153] The concentrations of insulin aspart and GLP-1 at each blood collection point were measured using an ELISA kit. PK parameters were analyzed using PKSolver 2.0 - Compartmental Analysis of plasma data after extravascular input of the model (two compartments).

[0154] Changes in aspart insulin concentration over time are as follows: Figure 10 As shown, the PK parameters are summarized in Table 4A.

[0155] Table 4A Aspart Insulin PK Parameters

[0156] T in the aspart insulin alone group max For approximately 45 minutes, the combined group's T max The time to peak concentration was approximately 32 minutes. The time to peak concentration after combined administration was 13 minutes earlier than with insulin aspart alone, indicating that insulin aspart has a faster onset of action in combined administration than when administered alone. The C-value of insulin aspart in combination administration... max and AUC 0-t These were 1.22 and 1.29 times higher than those achieved with insulin aspart alone, respectively.

[0157] Similarly, GLP-1 concentrations at each blood collection point were measured and analyzed in both the GLP-1 single and combined groups. Changes in GLP-1 concentrations over time were observed. Figure 11 As shown in the image.

[0158] Considering that the dose of GLP-1 in the GLP-1-only group was twice that in the combination group, the PK parameters were adjusted to the same dose of GLP-1 (10 μg / kg). The adjusted PK parameters are summarized in Table 4B: Table 4B GLP-1 PK Parameters

[0159] GLP-1 alone in GLP-1 T max For approximately 17 minutes, the T in the combined group GLP-1 max The time to peak concentration was approximately 14 minutes. In the combination group, the time to peak concentration was slightly shorter than in the GLP-1-only group. The corrected C1 for the combination group... max and AUC 0-t These are 1.24 and 1.09 times higher than those of the GLP-1-only group, respectively.

[0160] GLP-1 was prepared using benaglutide (2 mg / ml, acetate 10 mM, mannitol 30 mg / ml, phenol 2.2 mg / ml, propylene glycol 5 mg / ml, pH 4.0, Shanghai Benemae).

[0161] The regimen consisted of 100 U (3.5 mg / ml) of aspart insulin, 16 mg / ml of glycerol, 1.5 mg / ml of phenol, 1.72 mg / ml of m-cresol, 1.25 mg / ml of disodium hydrogen phosphate dihydrate, 0.58 mg / ml of sodium chloride, 19.6 μg / ml of zinc, pH 7.4 (Novo Nordisk).

[0162] The combination composition contains 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, 0.2% DDM, 40 mM NaCl, and phosphate buffer (PB).

[0163] In the description of this specification, the terms "one embodiment," "some embodiments," "example," "specific embodiment," or "some embodiments," etc., mean that a specific feature, structure, material, or feature is described in connection with an embodiment or example and is included in at least one embodiment or example of the invention. In this specification, the illustrative representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described can be combined in any suitable manner in any one or more embodiments or examples.

[0164] Although embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are illustrative and should not be construed as limiting the present invention. Those skilled in the art will not depart from the principles and objectives of the present invention. In such cases, within the scope of the present invention, the above embodiments may be changed, modified, substituted, and altered.

Claims

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

2. The pharmaceutical composition of claim 1, further comprising a second active ingredient comprising insulin, an insulin analog, and / or a derivative of the insulin analog, optionally, the insulin analog comprises at least one selected from insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, or insulin degludec, optionally, the pharmaceutical composition comprises 0.1-1 wt% of insulin or insulin analog or derivative of insulin analog, and optionally, 0.1-0.4 wt% of insulin or insulin analog or derivative of insulin analog, based on the total weight of the pharmaceutical composition.

3. The pharmaceutical composition of claim 1, wherein, the alkyl glycoside comprises a sugar group and an alkyl group connected by a linking group, wherein: the sugar group comprises at least one of 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 thio glycosidic bond, or an amide bond.

4. The pharmaceutical composition of claim 3, wherein, the alkyl chain comprises 10-14 carbon atoms.

5. The pharmaceutical composition of claim 3, wherein, the sugar group comprises maltose or sucrose.

6. The pharmaceutical composition of claim 1, wherein, the alkyl glycoside comprises at least one compound selected from n-dodecyl beta-D-maltoside (DDM), sucrose dodecanoate, n-dodecyl beta-D-maltoside, n-hexyl-beta-D-glucoside, n-heptyl-beta-D-glucoside, n-octyl-beta-D-glucoside, n-nonyl-beta-D-glucoside, n-decyl-beta-D-glucoside, 3-cyclohexyl-1-propyl-beta-D-glucoside, n-hexyl-beta-D-glucopyranoside, n-octyl-beta-D-maltoside, n-nonyl-beta-D-maltoside, n-decyl-beta-D-maltoside, cyclohexyl-methyl-beta-D-maltoside, 2-cyclohexyl-ethyl-beta-D-maltoside, 3-cyclohexyl-propyl-beta-D-maltoside, 4-cyclohexyl-butyl-beta-D-maltoside, or 5-cyclohexyl-pentyl-beta-D-maltoside, optionally, the concentration of the alkyl glycoside is about 0.05% wt to about 0.50% wt or about 0.1% wt to about 0.20% wt, based on the total weight of the pharmaceutical composition.

7. The pharmaceutical composition of claim 1, wherein, the GLP-1 receptor agonist is a peptide agonist.

8. The pharmaceutical composition of claim 7, wherein, the GLP-1 receptor agonist is also a GIP agonist.

9. The pharmaceutical composition according to claim 7, wherein, the GLP-1 receptor agonist comprises at least one of: a GLP-1, a GLP-1 analog, or a derivative of the GLP-1 analog; an extended GLP-1 fragment, or an analog or derivative thereof; and a truncated GLP-1 fragment, or an analog or derivative thereof.

10. The pharmaceutical composition of claim 7, wherein, the GLP-1 analog comprises exenatide-4.

11. The pharmaceutical composition of claim 1, wherein, the GLP-1 receptor agonist comprises at least one selected from liraglutide, exenatide, lixisenatide, albiglutide, benalfrataide, dulaglutide, semaglutide, cilarglutide, and taspoglutide.

12. The pharmaceutical composition of claim 1, wherein, the first active ingredient comprises or is benralizumab, and the second active ingredient comprises or is insulin glargine; the first active ingredient comprises or is benralizumab, and the second active ingredient comprises or is insulin aspart.

13. The pharmaceutical composition of any one of claims 1-12, wherein the pharmaceutical composition is a liquid pharmaceutical composition or a pre-mixed aqueous injectable pharmaceutical composition.

14. The pharmaceutical composition of claim 13, further comprising a bacteriostatic agent, optionally, the bacteriostatic agent comprises at least one compound selected from a phenolic compound, an aromatic alcohol compound, a phenoxyethanol, or a chlorinated alcohol compound.

15. The pharmaceutical composition of claim 14, wherein, the bacteriostatic agent comprises at least one of phenol, m-cresol, benzyl alcohol, phenoxyethanol, or chlorobutanol.

16. The pharmaceutical composition of claim 15, wherein, the GLP-1 receptor agonist is at a concentration of about 0.1 mg / mL to about 10 mg / mL.

17. The pharmaceutical composition of claim 15, wherein, the pH of the pharmaceutical composition is about 3.0 to about 9.0, optionally, the insulin analog is insulin glargine, and the pH of the pharmaceutical composition is about 3.0 to about 5.0, and optionally, the pH of the pharmaceutical composition is about 4.0 to about 4.5, optionally, the insulin analog is insulin aspart, and the pH of the pharmaceutical composition is about 7.0 to about 9.

0.

18. The pharmaceutical composition of claim 15, wherein, the pharmaceutical composition further comprises zinc ions, optionally, the zinc ions are at a concentration of about 1 pg / mL to about 100 pg / mL, about 10 pg / mL to about 50 pg / mL, about 20 pg / mL to about 40 pg / mL.

19. The pharmaceutical composition according to claim 1, wherein, the pharmaceutical composition further comprises a solubilizing agent, the solubilizing agent comprises at least one of glycerol, mannitol, propylene glycol, and combinations thereof, at a concentration of about 10 mg / mL to about 50 mg / mL based on the total volume of the pharmaceutical composition; preferably, the solubilizing agent is glycerol at a concentration of about 10 mg / mL to about 25 mg / mL based on the total volume of the pharmaceutical composition, or the pharmaceutical composition does not comprise any of polysorbate 20, polysorbate 80, poloxamer 188, hydroxypropyl beta-cyclodextrin, and amino acids, optionally, the surfactant comprises only alkylglycoside.

20. The pharmaceutical composition of claim 1, wherein, the pharmaceutical composition does not form foreign matter when subjected to an accelerated stability test at 25 °C for 3 months, and the peptide is not less than about 93%.

21. A method for improving the storage stability of a pharmaceutical composition comprising a GLP-1 receptor agonist, comprising mixing the pharmaceutical composition with one or more additives of alkylglycoside.

22. The method of claim 21, wherein, the pharmaceutical composition further comprises an insulin or an insulin analog or a derivative of the insulin analog, and optionally, the insulin analog comprises at least one of insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, and insulin degludec.

23. The method of claim 21, wherein, The alkyl glycoside includes at least one of n-dodecyl beta-D-maltoside (DDM), sucrose laurate, n-dodecyl beta-D-maltoside, n-hexyl-beta-D-glucoside, n-heptyl-beta-D-glucoside, n-octyl-beta-D-glucoside, n-nonyl-beta-D-glucoside, n-decyl-beta-D-glucoside, 3-cyclohexyl-1 -propyl-beta-D-glucoside, n-hexyl-beta-D-glucopyranoside, n-octyl-beta-D-maltoside, n-nonyl-beta-D-maltoside, n-decyl-beta-D-maltoside, cyclohexyl-methyl-beta-D-maltoside, 2-cyclohexyl-ethyl-beta-D-maltoside, 3-cyclohexyl-propyl-beta-D-maltoside, 4-cyclohexyl-butyl-beta-D-maltoside, or 5-cyclohexyl-pentyl-beta-D-maltoside.

24. Use of the pharmaceutical composition of any one of claims 1-20 in (i) the prevention and / or treatment of diabetes, and / or reduction of HbAlC; (ii) the delay or prevention of the progression of diabetes, the delay of the progression of impaired glucose tolerance to insulin requiring type 2 diabetes, the delay or prevention of insulin resistance and / or the delay of the progression of non-insulin requiring type 2 diabetes to insulin requiring type 2 diabetes; (iii) the improvement of beta-cell function, and / or the restoration of beta-cell glucose sensitivity; (iv) the prevention and / or treatment of eating disorders; the reduction of gastric motility; the delay of gastric emptying; (v) the prevention and / or treatment of complications of obesity; (vi) the prevention and / or treatment of complications of diabetes; and / or (vii) the prevention and / or treatment of cardiovascular diseases.

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