GLP-1 and GIP dual receptor agonist pharmaceutical composition and its use

Abstract

The present invention provides a pharmaceutical composition for a GLP-1 and GIP dual receptor agonist, which comprises a GLP-1 and GIP dual receptor agonist, a stabilizer, and a buffer salt. The present invention further provides a use of the pharmaceutical composition in the manufacture of a medicament for preventing and / or treating a metabolic disorder-related disease.

Description

[Technical Field]

[0001] This disclosure claims priority to a Chinese patent application with application number 202211167785.2, filed with the China Patent Office on September 23, 2022, which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to the pharmaceutical field, and specifically to a pharmaceutical composition of a GLP-1 and GIP dual receptor agonist and its use. [Background technology]

[0003] Glucose-dependent insulinotropic peptide (GIP) is a 42-amino acid gastrointestinal regulatory peptide that plays a physiological role in glucose homeostasis by stimulating insulin secretion from pancreatic β cells and protecting pancreatic β cells in the presence of glucose. Glucagon-like peptide-1 (GLP-1) is a 37-amino acid peptide that stimulates insulin secretion, protects pancreatic β cells, suppresses glucagon secretion, gastric emptying, and food intake, and induces weight loss. GIP and GLP-1 are secreted by K cells and L cells in the small intestinal epithelium, respectively, and are known as incretins. Incretin receptor signaling plays a physiologically important role in glucose homeostasis.

[0004] GIP and GLP-1 exert their physiological effects by binding to their specific receptors, such as the GIP (GIPR) receptor and the GLP-1 receptor (GLP-1R), respectively. Research has shown that the cooperative action of GLP-1R / GIPR exerts a synergistic blood glucose lowering effect, and GIP and GLP-1 dual receptor agonists may be able to achieve better blood glucose lowering effects and stimulate insulin secretion. Therefore, the development of GLP-1 and GIP dual receptor agonists and pharmaceutical compositions thereof with novel structures and good therapeutic effects has been a research hotspot in the field of metabolic disease treatment, such as diabetes and obesity.

[0005] Currently, polypeptide-based drugs are often stored and used in the form of liquid formulations, but polypeptide active ingredients are easily affected by the external environment, which can cause chemical changes such as hydrolysis and oxidation, and the resulting impurities have a significant impact on the efficacy and safety of the drug.

[0006] GLP-1 and GIP dual receptor agonists are generally linear peptides, and the pH of the formulation must be close to human physiological pH. Therefore, the pH of the formulation is generally controlled at around 7–8. Polypeptide drugs are susceptible to microbial degradation under these pH conditions. Therefore, the addition of a bactericide (e.g., phenol) to the formulation is generally considered to improve product stability. For example, commercially available hypoglycemic drugs, such as semaglutide (Ozempic), contain 8.25 mg of phenol as a bactericide; exenatide contain 2.64 mg of m-cresol as a bactericide; liraglutide (Victozza) contain 16.5 mg of phenol as a bactericide; and lixisenatide (Adlyxin) contain 8.1 mg of m-cresol as a bactericide. However, because injectable drugs directly enter and circulate in the human body, the use of bactericides increases drug safety risks. National regulatory authorities have issued official documents urging the use of preservatives to be minimized when selecting insulin-based products for formulation. Therefore, solving the problem of antiseptic-free formulations of polypeptide hypoglycemic drugs is also a technical challenge in the field, and the development of preservative / antiseptic-free GLP-1 and GIP dual receptor agonist formulations has very good application prospects.

[0007] Polypeptide drugs are formed by linking multiple amino acids together through amide bonds formed by condensation reactions. Because polypeptide drugs have both electron-rich and electron-withdrawing groups, intermolecular attraction between molecules easily occurs during the preparation of drug formulations, resulting in polymerization of drug molecules. Once polymerized, polypeptide drugs lose their efficacy. Drug polymerization can also cause quality problems, such as cloudiness or discoloration of the injection. Therefore, the issue of drug polymerization during the preparation of polypeptide drug formulations has been a technical challenge in this field.

[0008] In summary, the development of a formulation method for a pharmaceutical composition of a GLP-1 and GIP dual receptor agonist that is suitable for long-term storage and has stable efficacy is a problem to be solved in the art. Summary of the Invention [Problem to be solved by the invention]

[0009] In order to develop a GLP-1 and GIP dual receptor agonist with a novel structure that can effectively treat obesity, diabetes, and their complications, the present applicants have proposed a GLP-1 and GIP dual receptor agonist in a patent application with application number 202210294984.3 and title "GIP and GLP-1 Dual Receptor Agonist, Pharmaceutical Composition, and Use." The dual receptor agonist is a dual receptor agonist for glucose-dependent incretin polypeptides GLP-1 and GIP independently developed by the present applicants, and its clinical indication is type 2 diabetes. The drug is a linear peptide containing 40 amino acids and has the effect of activating both GLP-1R and GIPPR. Preclinical studies have shown that this dual receptor agonist has strong agonistic effects on both GLP-1 and GIP receptors, with activity 2-3 times stronger than that of tirzepatide, and its side chain consists of a single fatty acid chain linked to two AEEEA chains, the main function of which is to bind to serum albumin (albumin contains basic residues), thereby prolonging its half-life and achieving long-term action. Preclinical pharmacokinetic studies have shown that the binding rates of this dual receptor agonist to various plasma proteins all exceed 99.4%, and its in vivo half-lives in rats and cynomolgus monkeys are 14 hours and 40 hours, respectively.

[0010] For the GLP-1 and GIP dual receptor agonists, there is currently no pharmaceutical formulation suitable for long-term storage and ensuring stable efficacy. In response to this, the present disclosure aims to provide a pharmaceutical composition for a GLP-1 and GIP dual receptor agonist, which can maintain the stability of the GLP-1 and GIP dual receptor agonist in a liquid formulation by adding excipients such as stabilizers and buffer salts, and effectively prevent its physical and chemical degradation, and uses thereof. [Means for solving the problem]

[0011] According to a first aspect of the present disclosure, there is provided a pharmaceutical composition comprising a GLP-1 and GIP dual receptor agonist, a stabilizer, and a buffer salt, wherein the GLP-1 and GIP dual receptor agonist is a compound of Formula I or a pharmaceutically acceptable salt, ester, solvate, optical isomer, tautomer, isotopic marker, or prodrug thereof. [ka]

[0012] Furthermore, the pharmaceutical composition is a liquid formulation, and preferably, the pharmaceutical composition is an injection.

[0013] Furthermore, the concentration of the GLP-1 and GIP dual receptor agonist in the pharmaceutical composition is 2 to 36 mg / mL, and preferably 5 to 30 mg / mL.

[0014] Furthermore, in the pharmaceutical composition, the stabilizer is a polyol, preferably, the stabilizer is propylene glycol, mannitol, or glycerol, more preferably, the stabilizer is propylene glycol.

[0015] Furthermore, the concentration of the stabilizer in the pharmaceutical composition is 10 to 20 mg / mL.

[0016] Further, in the pharmaceutical composition, the buffer salt is selected from hydrogen phosphate, hydrogen phosphate hydrate, citrate, or citrate hydrate, preferably, the buffer salt is disodium hydrogen phosphate dodecahydrate or sodium citrate dihydrate, more preferably, the buffer salt is disodium hydrogen phosphate dodecahydrate.

[0017] Furthermore, the concentration of the buffer salt in the pharmaceutical composition is 7 to 23 mM.

[0018] Furthermore, the pH value of the pharmaceutical composition is 6.5 to 8.5, preferably 7.0 to 8.4, preferably 7.2 to 8.2, and more preferably 7.2 to 7.6.

[0019] Furthermore, the pharmaceutical composition further contains a pH adjuster and water, the pH adjuster being hydrochloric acid and / or sodium hydroxide, the pH value of the pharmaceutical composition being adjusted by the pH adjuster, and the water being preferably purified water and / or water for injection.

[0020] Furthermore, the pharmaceutical composition does not contain a bactericide.

[0021] According to a second aspect of the present invention, there is provided a pharmaceutical composition comprising a GLP-1 and GIP dual receptor agonist represented by formula I, a stabilizer, and a buffer salt, wherein the stabilizer is propylene glycol and the buffer salt is disodium hydrogen phosphate dodecahydrate, the pH value of the pharmaceutical composition is 7.0 to 8.4, and in the pharmaceutical composition, the concentration of the GLP-1 and GIP dual receptor agonist represented by formula I is 5 to 30 mg / mL, the concentration of the propylene glycol is 10 to 20 mg / mL, and the concentration of the disodium hydrogen phosphate dodecahydrate is 7 to 23 mM; [ka] The pharmaceutical composition is an injection.

[0022] Furthermore, the pH value of the pharmaceutical composition is 7.2 to 8.2, and the concentration of disodium hydrogen phosphate dodecahydrate in the pharmaceutical composition is 10 to 20 mM.

[0023] Furthermore, the pH value of the pharmaceutical composition is 7.2 to 7.6, the concentration of the GLP-1 and GIP dual receptor agonist represented by formula I is 5 to 25 mg / mL, the concentration of the propylene glycol is 14 mg / mL, and the concentration of the disodium hydrogen phosphate dodecahydrate is 10 mM.

[0024] Furthermore, the pharmaceutical composition further comprises a pH adjuster and water, preferably the pH adjuster is hydrochloric acid and / or sodium hydroxide, and preferably the water is purified water and / or water for injection.

[0025] According to a third aspect of the present disclosure, there is provided a method for producing the pharmaceutical composition according to the first or second aspect, comprising the steps of: formulating a stabilizer, a buffer salt, and a GLP-1 and GIP dual receptor agonist into a solution; and adjusting the pH value of the solution to a target value to obtain the pharmaceutical composition.

[0026] The method further includes the steps of dissolving a stabilizer and a buffer salt in a solvent, adding a GLP-1 and GIP dual receptor agonist to prepare a solution, adding a pH adjuster to completely dissolve the GLP-1 and GIP dual receptor agonist, and further adjusting the pH of the solution to a target value using the pH adjuster to obtain a pharmaceutical composition.

[0027] According to a fourth aspect of the present disclosure, there is provided a use of the pharmaceutical composition according to the first or second aspect in the manufacture of a medicament for preventing and / or treating a metabolic disorder-related disease, preferably wherein the metabolic disorder-related disease is diabetes, diabetic complications, obesity or obesity complications. [Effects of the Invention]

[0028] The present disclosure adds excipients such as stabilizers and buffer salts to the provided pharmaceutical compositions to ensure that the GLP-1 and GIP dual receptor agonist as an active pharmaceutical ingredient maintains good stability in liquid formulations, and avoids quality issues such as ineffectiveness of the drug due to polymerization between the GLP-1 and GIP dual receptor agonist as a polypeptide drug, ultimately realizing long-term storage of the pharmaceutical composition and showing important application prospects in the prevention and treatment of metabolic disorder-related diseases such as diabetes or diabetic complications, and obesity or obesity complications.

[0029] The present disclosure utilizes a pH adjusting agent to control the pH value of the pharmaceutical composition within a specific range, which is more advantageous for the complete dissolution of the GLP-1 and GIP dual receptor agonist during the preparation of the pharmaceutical composition, and ensures the stability of the GLP-1 and GIP dual receptor agonist as an active pharmaceutical ingredient during storage of the composition, avoiding its decomposition.

[0030] The present disclosure utilizes the selection of specific stabilizers to significantly increase the solubility of GLP-1 and GIP dual receptor agonists (e.g., compounds of Formula I) while allowing them to retain their intact structure and biological activity during storage.

[0031] The present disclosure can ensure the stability of the pH value of the pharmaceutical composition during storage by selecting a specific buffer salt, and does not affect the quality and stability of the active pharmaceutical ingredient.

[0032] The method for producing a pharmaceutical composition according to the present disclosure is easy to operate, has low production costs, is favorable for scalable production, and supports a wide range of applications of drug formulations.

[0033] The pharmaceutical composition according to the present disclosure can effectively prevent and / or treat diseases associated with metabolic disorders such as diabetes, diabetic complications, obesity or obesity complications, and is convenient to use. [Brief explanation of the drawings]

[0034] [Figure 1]10 is a graph dedicated to the validation of the multimer detection method. [Figure 2] 1 is a graph showing the detection of multimers in an injection of a compound of formula I. DETAILED DESCRIPTION OF THE INVENTION

[0035] <Definition> Unless stated to the contrary, terms used in this disclosure have the following meanings.

[0036] In the claims and / or specification of the present invention, the words "a" or "an" or "the" can refer to "one," but can also mean "one or more," "at least one," and "one or more."

[0037] As used in the claims and the specification, the words "comprise," "have," "comprehensive," or "containing" are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. At the same time, "comprise," "have," "comprehensive," or "containing" can also refer to closed elements or method steps, excluding additional, unrecited elements or method steps.

[0038] In this disclosure, the term "about" indicates that a value includes the standard deviation of error for the device or method being employed to determine the value.

[0039] In the present disclosure, the term "agonist" refers to a substance (ligand) that activates signal transduction through a target receptor type. Illustratively, when the GLP-1 receptor is the target receptor, the agonist has activation activity of the GLP-1 receptor, such as a GLP-1 polypeptide or an analog thereof.

[0040] In the present disclosure, the term "treatment" refers to alleviating the symptoms of a disease compared to the absence of such contact by contacting (e.g., administering) a pharmaceutical composition of the present disclosure to a subject after the subject has contracted the disease, and does not mean that the symptoms of the disease need to be completely suppressed. Contracting a disease refers to the appearance of disease symptoms in the body.

[0041] In the present disclosure, the term "prevention" refers to reducing symptoms after contracting a disease compared to the absence of contact by contacting (e.g., administering) a pharmaceutical composition of the present disclosure to a subject before the subject is affected with the disease, and does not mean that symptoms of the disease need to be completely suppressed.

[0042] In this disclosure, the terms "individual," "patient," or "subject" include mammals, including, but not limited to, domestic animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats).

[0043] In the present disclosure, the term "therapeutically effective amount" refers to an amount that effectively achieves a desired therapeutic result, at the necessary dosage and for the necessary period of time. The therapeutically effective amount of a compound or pharmaceutical composition of the present disclosure may vary depending on various factors, such as the disease state, the age, sex, and weight of the individual, and the ability of the immune adjuvant or pharmaceutical composition to elicit a desired response in the individual.

[0044] In this disclosure, the term "pharmaceutically acceptable salt" refers to a salt prepared from a compound of this disclosure with a relatively non-toxic acid or base. When a compound of this disclosure contains a relatively acidic functional group (e.g., a carboxyl group or a sulfonic acid group), a base addition salt can be obtained by contacting the free form with a sufficient amount of base in a pure solution or in a suitable inert solvent. Non-limiting examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, organic amine salts, or similar salts. When a compound of this disclosure contains a relatively basic functional group (e.g., an amino or guanidino group), an acid addition salt can be obtained by contacting the free form with a sufficient amount of acid in a pure solution or in a suitable inert solvent. Non-limiting examples of pharmaceutically acceptable acid addition salts include inorganic acid salts (e.g., hydrochloride, hydrobromide, hydroiodide, nitrate, carbonate, bicarbonate, phosphate, monohydrogenphosphate, dihydrogenphosphate, phosphite, sulfate, hydrogensulfate, etc.), organic acid salts (e.g., acetate, propionate, isobutyrate, malonate, succinate, suberate, maleate, fumarate, citrate, tartrate, lactate, mandelate, benzoate, phthalate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, glucuronic acid, etc.), and amino acid salts (e.g., arginine salt, etc.). Specific forms of pharmaceutically acceptable salts may be further described in Berge et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66: 1-19).

[0045] In the present disclosure, the term "metabolic disorder" can be a carbohydrate metabolic disorder such as diabetes, diabetic complications, obesity, obesity complications, etc. Since the relationship between obesity and diabetes and blood glucose metabolism is well-known, these disorders do not necessarily have to be, but can be separate or mutually exclusive. In some embodiments, diabetes or diabetic complications include insulin resistance, impaired glucose tolerance, elevated fasting blood glucose, prediabetes, type I diabetes, type II diabetes, gestational diabetes mellitus, dyslipidemia, or combinations thereof. In some embodiments, obesity complications include obesity-related inflammation, obesity-related gallbladder disease, or sleep apnea induced by obesity, or may be selected from associated diseases such as atherosclerotic dyslipidemia, dyslipidemia, elevated blood pressure, hypertension, prethrombotic state and inflammation-induced state, or combinations thereof.

[0046] The pharmaceutical composition of the present disclosure has excellent dual agonistic activity of GIPR and GLP-1R, can effectively lower blood glucose, control the weight gain of type 2 diabetic model mice, and is used for the prevention and / or treatment of metabolic disorder diseases, and has good clinical application and pharmaceutical uses.

[0047] <Dual Receptor Agonist of GLP-1 and GIP> In the present disclosure, the dual receptor agonist of GLP-1 and GIP refers to a substance that targets the GLP-1 receptor and the GIP receptor and has simultaneous activation activity on GLP-1R and GIPR.

[0048] In some specific embodiments, the dual receptor agonist of GLP-1 and GIP described in the present disclosure is a compound represented by Formula I or a pharmaceutically acceptable salt, ester, solvate, optical isomer, tautomer, isotope marker or prodrug thereof.

Chemical Structure

[0049] The present disclosure does not particularly limit the content of the drug active ingredient in the pharmaceutical composition, and it may be determined by those skilled in the art according to actual needs.

[0050] In some embodiments, the concentration of the GLP-1 and GIP dual receptor agonist described herein is 2 to 36 mg / mL, e.g., 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, 21 mg / mL, 22 mg / mL, 23 mg / mL, 24 mg / mL, 25 mg / mL, 26 mg / mL, 27 mg / mL, 28 mg / mL, 29 mg / mL, 30 mg / mL, 31 mg / mL, 32 mg / mL, 33 mg / mL, 34 mg / mL, 35 mg / mL, 36 mg / mL, etc., preferably 5 to 30 mg / mL.

[0051] <Stabilizer> In the present disclosure, stabilizers refer to substances that stabilize the structure and biological activity of polypeptide substances in solution. Particularly for polypeptide formulations, screening for suitable stabilizers is the main means for preventing the physical and chemical degradation of polypeptide drug molecules in liquid formulations. Selecting suitable stabilizers can improve the stability of polypeptide molecules by binding the hydrophobic portion of polypeptide molecules, increasing the viscosity of the solution, and affecting the folding state of polypeptide molecules.

[0052] The present disclosure does not particularly limit the specific type of stabilizer, which may be determined by those skilled in the art according to actual needs.

[0053] In some embodiments, the stabilizers described herein are selected from polyols.

[0054] In some specific embodiments, the stabilizer described herein is propylene glycol, mannitol, or glycerol.

[0055] Based on the results of the formulation screening test, combined with the results of the stability test, the present disclosure preferentially selects propylene glycol as a stabilizer, which can effectively maintain the structure and biological activity of the active ingredient, and can also stabilize the pH value of the pharmaceutical composition during storage.

[0056] The present disclosure does not particularly limit the dosage of the stabilizer, which may be determined by those skilled in the art according to actual needs.

[0057] In some embodiments, the concentration of the stabilizer described herein is 10-20 mg / mL, e.g., 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, etc., preferably 14 mg / mL.

[0058] It is worth noting that in polypeptide-based formulations, sodium chloride is known to maintain electrolyte balance and stabilize polypeptide structure, and it has been used as a stabilizer in various conventional polypeptide-based formulations.However, the inventors of the present disclosure have found that when sodium chloride is used instead of propylene glycol (for example, 2 mL of pharmaceutical composition contains 40.0 mg of the compound of formula I, 3.58 mg of disodium hydrogen phosphate dodecahydrate, and 16.4 mg of sodium chloride, and the pH value is adjusted to 7 using a pH adjuster, and the remaining amount is made up with water for injection), the solubility of the compound of formula I is only 0.15 mg / mL, which is far below the required level. When propylene glycol is used as a stabilizer (e.g., 1 mL of the pharmaceutical composition contains 20.0 mg of the compound of Formula I, 3.58 mg of disodium hydrogen phosphate dodecahydrate, and 14 mg of propylene glycol, and the pH is adjusted to 7 using a pH adjuster, followed by the remaining volume being made up with water for injection), the solubility of the compound of Formula I can reach 30 mg / mL or more, satisfying the solubility requirements of the active ingredient. Therefore, in some preferred embodiments of the present disclosure, the pharmaceutical composition does not contain sodium chloride.

[0059] <pH Value and pH Regulator> The pH value has a great impact on the stability of the polypeptide drug GLP-1 and the dual receptor agonist of GIP, and also affects the dissolution status of the main drug in the pharmaceutical composition formulation. A relatively acidic environment is unfavorable for the dissolution of the dual receptor agonist of GLP-1 and GIP according to the present disclosure. However, with the increase of pH, during product storage, the higher the reduction rate of purity, the more unfavorable the relatively basic conditions are for the stability of the product.

[0060] In some embodiments, the pH value of the pharmaceutical composition of the present disclosure is 6.5 - 8.5, such as 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7._{7}, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, etc., preferably 7.0 - 8.4, more preferably 7.2 - 8.2, and even more preferably 7.2 - 7.6. <(

[0061] A reasonable pH value range not only guarantees the purity and stability of the pharmaceutical composition of the present disclosure during storage, but also takes into account the pH fluctuations during the drug storage period, thereby guaranteeing the stability of the whole process from the production to the use of the pharmaceutical composition. [[ID=(13]]

[0062] In some embodiments, the present disclosure uses a pH regulator to adjust the pH value of the pharmaceutical composition.

[0063] In some specific embodiments, the pH regulator described in the present disclosure is hydrochloric acid and / or sodium hydroxide. Hydrochloric acid is a transparent, colorless, fuming hydrochloric acid solution with a pungent odor and is often used as an oxidizing agent in pharmaceutical preparations. Sodium hydroxide is a molten white dry particle, lump, rod or flake, generally recognized as non-toxic at low concentrations, and is widely used to adjust the pH value of solutions in pharmaceutical preparations. In some more specific embodiments, the pH regulator described in the present disclosure may be hydrochloric acid at a certain concentration and / or sodium hydroxide solution at a certain concentration.

[0064] The present disclosure does not limit the concentration of the pH adjuster, and those skilled in the art may adjust it according to actual production needs. For example, a low concentration pH adjuster, for example, a pH adjuster with a concentration of less than 3M, may be selected, or a high concentration pH adjuster, for example, a pH adjuster with a concentration of more than 3M, may be selected, as long as neither of them has an adverse effect on the active pharmaceutical ingredient. The present disclosure preferably uses 0.1M hydrochloric acid and / or 0.1M sodium hydroxide solution.

[0065] <Buffer salts> In the present disclosure, buffer salt refers to a substance that stabilizes the acidity and alkalinity of a solution. Particularly for polypeptide-based formulations, screening for an appropriate buffer salt system is an important means to prevent the physical and chemical degradation of polypeptide drug molecules in liquid formulations. Selecting an appropriate buffer salt system can maintain the stability of the pH value of the polypeptide solution and improve the stability of the polypeptide by affecting electrostatic effects.

[0066] The present disclosure does not particularly limit the specific type of buffer salt, which may be determined by those skilled in the art according to actual needs.

[0067] In some embodiments, the buffer salt described herein is selected from hydrogen phosphate, hydrogen phosphate hydrate, citrate, or citrate hydrate.

[0068] In some specific embodiments, the buffer salt described herein is disodium hydrogen phosphate dodecahydrate or sodium citrate dihydrate.

[0069] In some preferred embodiments, the buffer salt described herein is disodium hydrogen phosphate dodecahydrate, which not only helps stabilize the pH value of the pharmaceutical composition of the present disclosure during storage, but also helps stabilize the purity of the pharmaceutical composition during storage.

[0070] Based on the results of the formulation screening test, combined with the stability test data, the present disclosure preferentially selects disodium hydrogen phosphate dodecahydrate as the buffer salt, which has stable chemical properties, does not affect the quality and stability of the active ingredient, and has good effect on maintaining the stability of the pH value of the formulation.

[0071] The present disclosure does not specifically limit the dosage of the buffer salts, which may be determined by those skilled in the art according to actual needs.

[0072] In some embodiments, the concentration of the buffer salt described herein is 7-23 mM, e.g., 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, etc., preferably 7-20 mM, more preferably 7-15 mM.

[0073] Based on the results of the formulation screening test and taking into consideration the degree of risk in subsequent clinical application of the pharmaceutical composition as an injection formulation or the like, the present disclosure preferentially selects a low concentration of buffer salt, for example, a concentration of 10 mM.

[0074] <Fungicide> The GLP-1 and GIP dual receptor agonist described in the present disclosure is a linear peptide containing 40 amino acids, and the pH value of the formulation is initially set to between 7.2 and 8.2, making it susceptible to microbial degradation. Therefore, it is generally considered to add a bactericide (e.g., phenol) to the formulation to improve the stability of the product.

[0075] Furthermore, because polypeptide drugs are easily decomposed by microorganisms, adding a bactericide to a polypeptide hypoglycemic drug is a conventional technical means in the art. For example, commercially available hypoglycemic drug products, semaglutide (OZEMPIC), contain 8.25 mg of phenol as a bactericide, exenatide contain 2.64 mg of m-cresol as a bactericide, liraglutide (VICTOZA) contain 16.5 mg of phenol as a bactericide, and lixisenatide (ADLYXIN) contain 8.1 mg of m-cresol as a bactericide.

[0076] However, after a long period of research and investigation, the inventors of the present disclosure have discovered that the safety of clinical drugs can be ensured without adding a bactericide, and risk factors during application of pharmaceutical compositions can be further reduced.

[0077] If the composition does not contain a bactericide, it is more advantageous to prevent or reduce risks during clinical use of the drug formulation, reduce the production costs of the pharmaceutical composition to a certain extent, and be more advantageous for large-scale production and application of the drug.

[0078] <Pharmaceutical Composition> The pharmaceutical compositions of the present disclosure comprise a GLP-1 and GIP dual receptor agonist, a stabilizer, and a buffer salt.

[0079] In some embodiments, the pharmaceutical compositions described herein further comprise a pH adjusting agent and a solvent.

[0080] In some specific embodiments, the pharmaceutical compositions described herein consist of a GLP-1 and GIP dual receptor agonist, a stabilizer, a buffer salt, a pH adjusting agent, and a solvent.

[0081] The pharmaceutical composition according to the present disclosure exhibits good activation effects on GLP-1R and GIPR in vivo. The explanation of each component in the pharmaceutical composition is as described above.

[0082] In some embodiments, the pharmaceutical composition of the present disclosure is a liquid formulation.

[0083] In some specific embodiments, the pharmaceutical compositions described herein are injectable.

[0084] In some more specific embodiments, the solvent used by the pharmaceutical compositions of the present disclosure as liquid formulations is water, preferably purified water or water for injection.

[0085] The present disclosure further provides a method for preparing the pharmaceutical composition, which includes the steps of formulating a stabilizer, a buffer salt, and a GLP-1 and GIP dual receptor agonist into a solution, and adjusting the pH value of the solution to a target value to obtain the pharmaceutical composition.

[0086] In some specific embodiments, the method for preparing the pharmaceutical composition includes dissolving a stabilizer and a buffer salt in a solvent, adding a GLP-1 and GIP dual receptor agonist, adding a pH adjuster to completely dissolve the GLP-1 and GIP dual receptor agonist, and then adjusting the pH of the solution to a target value using the pH adjuster to obtain the pharmaceutical composition.

[0087] In some more specific embodiments, the method for preparing the pharmaceutical composition includes dissolving the prescribed amounts of disodium hydrogen phosphate dodecahydrate and propylene glycol in purified water or water for injection, adding the prescribed amount of GLP-1 and GIP dual receptor agonist, adding sodium hydroxide to completely dissolve the GLP-1 and GIP dual receptor agonist, adjusting the pH of the solution to 6.5 to 8.5 with hydrochloric acid and / or sodium hydroxide, and then adding purified water or water for injection to make the final volume to obtain the pharmaceutical composition. The purpose of adding sodium hydroxide is to appropriately increase the pH of the solution, which is more beneficial for dissolving the GLP-1 and GIP dual receptor agonist.

[0088] In the present disclosure, the route of administration can be varied or adjusted in any suitable manner to meet the needs of the properties of the drug, the convenience of the patient and medical personnel, and other relevant factors.

[0089] <Medicinal Uses of the Pharmaceutical Composition> The pharmaceutical composition of the present disclosure can be used to prevent and / or treat a metabolic disorder-related disease, and preferably, the metabolic disorder-related disease is diabetes, diabetic complications, obesity, or obesity complications.

[0090] The pharmaceutical composition of the present disclosure can further be used to prepare a drug for preventing and / or treating a metabolic disorder-related disease, and preferably, the metabolic disorder-related disease is diabetes, diabetic complications, obesity, or obesity complications.

[0091] The present disclosure further provides a method for preventing and / or treating a metabolic disorder-associated disease, which comprises administering to a subject a prophylactically and / or therapeutically effective amount of a pharmaceutical composition described in the present disclosure, preferably, the metabolic disorder-associated disease is diabetes, diabetic complications, obesity, or obesity complications.

[0092] Example In order to make the objectives and technical solutions of the present disclosure clearer, the embodiments of the present disclosure will be described in detail below with reference to examples. However, those skilled in the art should understand that the following examples are only used to illustrate the present disclosure and should not be considered to limit the scope of the present disclosure.

[0093] The reagents and instruments used in the examples are all commercially available products. Unless specific conditions are specified, the experiments are carried out according to conventional conditions or the conditions suggested by the manufacturer.

[0094] Example 1: Formulation Screening In consideration of the dosage form characteristics of this product as an injection, the concentration and type of buffer salts in the formulation, the type of stabilizer, and the pH range of the drug solution were currently screened. The compound of formula I used in the formulation study was self-prepared by the applicants, and all other materials used were commercially available.

[0095] (1) Screening of stabilizer types Stabilizers can improve the stability of polypeptide formulations by binding hydrophobic moieties to polypeptide molecules, increasing solution viscosity, and affecting the folding state of polypeptide molecules. To screen for suitable stabilizers, formulations were prepared using propylene glycol, mannitol, and glycerol. The resulting samples were then stored at 40°C and 2-8°C, respectively, to examine their high-temperature and long-term stability. Samples were taken at different time points to determine their properties, pH, and purity, and the impact of different stabilizers on the quality of the drug solution was examined. The formulation information is shown in Table 1, and the results are shown in Table 2.

[0096] [Table 1]

[0097] [Table 2]

[0098] The stabilized samples were all relatively stable at 2-8°C, and after one month, there were no significant changes in the properties, pH, purity, or maximum single impurity. After 10 days at a high temperature of 40°C, the purity of the samples decreased, but there were no significant changes in the properties or maximum single impurity. Surprisingly, however, the formulations using propylene glycol as the stabilizer showed a pH change of less than 3% under high-temperature accelerated stability conditions, demonstrating that the formulations using propylene glycol as the stabilizer achieved an unexpected technical advantage in maintaining pH stability. The formulations using mannitol and glycerol as stabilizers showed a significant pH change under high-temperature accelerated stability conditions, dropping by 0.7-0.8, with a clear tendency toward pH decline. Therefore, propylene glycol was the preferred stabilizer for this product.

[0099] (2) pH range screening pH is an important quality attribute of polypeptide drug formulations, directly affecting the solubility, stability, and aggregation state of polypeptide molecules. This study focused on the pH range of 5.0 to 8.4, focusing on the solubility of the drug substance and the stability of the product. The formulation information is shown in Table 3, and the results are shown in Table 4.

[0100] [Table 3]

[0101] [Table 4]

[0102] The results showed that when the pH was between 5.0 and 6.0, the solubility of the drug substance could not meet the drug formulation needs of this product, and the higher the pH, the better the dissolution of the drug substance. Within the target pH range of 7.4 to 8.4, the samples showed a greater decrease in purity under high temperature and accelerated conditions, i.e., relatively basic conditions are unfavorable to the stability of the product. However, from the perspective of decomposition, the decrease in product purity was still acceptable under the target pH of 8.4, and there were no significant changes in other considerations (properties, pH) for each sample.

[0103] (3) Screening of buffer salt concentrations Generally, pH and ionic strength have a significant impact on the stability and solubility of polypeptides. Ensuring the solubility of polypeptide drugs and improving their stability and safety has been a technical problem that polypeptide drug formulations must solve. A suitable buffer salt concentration is useful for solving this technical problem. Therefore, to screen for a suitable buffer salt concentration, sodium citrate dihydrate with concentrations of 10mM, 15mM, and 20mM was used to prepare formulations, and the prepared samples were stored at 40°C and 25°C, respectively, to examine the related indicators at high temperature and long-term. The specific formulation information is shown in Table 5, and the results are shown in Table 6.

[0104] [Table 5]

[0105] [Table 6]

[0106] The results showed that after leaving injection samples formulated with different buffer salt concentrations at 25°C for just one month, the purity of the 10 mM buffer salt formulation was slightly better than that of the 15 mM and 20 mM buffer salt formulations, but there were no significant differences in other considerations, indicating that buffer salt concentrations of 10 mM to 20 mM could all meet the requirements. However, considering that the higher the dosage of excipients in an injection, the greater the risk of clinical application of the formulation, the lowest concentration of 10 mM was selected as the buffer salt concentration for the injection of the present disclosure to further ensure the safety of the formulation.

[0107] (4) Screening of buffer salt systems The selection of a buffer system is also crucial for improving formulation stability. To screen for suitable buffer salts, formulations were prepared using sodium citrate dihydrate and disodium hydrogen phosphate dodecahydrate. The resulting samples were then stored at 40°C and 2-8°C, respectively, to examine the associated indicators for high temperature and long-term stability. Specific formulation information is shown in Table 7, and the results are shown in Table 8.

[0108] [Table 7]

[0109] [Table 8]

[0110] The results showed that both disodium hydrogen phosphate dodecahydrate and sodium citrate can be used as buffer salts for the injection of the present disclosure, and after standing at a high temperature of 40°C for 10 days, the purity of the phosphate formulation was slightly better than that of the citrate formulation, and disodium hydrogen phosphate dodecahydrate was preferably used as the buffer salt.

[0111] (5) Further confirmation of pH range The initial pH range confirmation results showed that a neutral pH value is beneficial for product stability, and the closer the pH is to physiological pH, the less irritating the product will be for subcutaneous administration. Therefore, a study using disodium hydrogen phosphate dodecahydrate as the buffer salt was designed to examine a narrower range around pH 7.4 to further confirm the optimal pH value of the product. The prescribing information is shown in Table 9, and the results are shown in Table 10.

[0112] [Table 9]

[0113] [Table 10]

[0114] The results showed that when the pH value was 7.2-7.6, after the sample was left at a high temperature of 40°C for 10 days, there was a slight increase in total impurities and polymers, but there was no significant change in the properties, pH value, maximum single impurity, or RRT of 1.04. After the sample was left under long-term conditions for 1 month, there was no significant change in any of the observation items, and there was no significant difference between samples with different pH values. Furthermore, even though no bactericide was added to the formulation of the present disclosure, it was still possible to control impurities within a range with no significant change, resulting in an unexpected technical effect.

[0115] In other embodiments of the present disclosure, the concentration of the active pharmaceutical ingredient (API) can be 5 mg / mL, 10 mg / mL, 15 mg / mL, 25 mg / mL, or 30 mg / mL, while the concentrations of other excipients remain the same. It can be understood that those skilled in the art can adjust the API concentration according to the specific conditions of patients, all of which are within the scope of protection of the present disclosure.

[0116] In the present disclosure, the purified water in the above formulations 1 to 16 may be replaced with water for injection, and the experimental result data obtained using the water for injection formulation after the replacement was not significantly different from the experimental result data using the above purified water formulation.

[0117] Example 2: Preparation of a pharmaceutical composition of a GLP-1 and GIP dual receptor agonist A composition was prepared using Formulation 15 as an example. 71.6 mg of disodium hydrogen phosphate dodecahydrate (buffer salt) and 280 mg of propylene glycol (for injection) (stabilizer) were dissolved in purified water, 400 mg of the compound of Formula I was added, and 0.1 M sodium hydroxide solution (pH adjuster) was added simultaneously. The mixture was stirred until the raw materials were dissolved, and the pH was adjusted to a target pH of 7.4 with 0.1 M hydrochloric acid or 0.1 M sodium hydroxide solution. Finally, the mixture was diluted to 20 mL with purified water. Other formulations were based on this manufacturing method.

[0118] Example 3: Detection of multimeric impurities in GLP-1 and GIP dual receptor agonist pharmaceutical compositions 1. Method for detecting multimers Research content: Interference of blank solution and blank excipients on multimer detection.

[0119] Expected requirements: Blank solutions and blank excipients should not interfere with the detection of multimers.

[0120] Testing process: (1) Blank solution: solvent.

[0121] (2) Blank excipient solution: obtained by taking blank excipient.

[0122] (3) System suitability solution: Approximately 10 mg of the compound of formula I was weighed accurately and placed in a vial. 5 ml of solvent was added and dissolved by ultrasonic waves. The solution was left at 60°C for 4 hours.

[0123] (4) Test solution: obtained by taking the GLP-1 and GIP dual receptor agonist pharmaceutical composition (compound of formula I injection).

[0124] 20 μL of the system suitability solution and 2 μL of the blank solution, blank excipient solution, and test article solution were precisely weighed and injected into the liquid chromatograph, and the chromatograms were recorded.

[0125] Test results: The blank solution and blank excipient did not interfere with the detection of the multimer, and the multimer and the compound of formula I showed sequential peaks in the system suitability solution chromatogram. The specific results are shown in Table 11 and Figure 1.

[0126] [Table 11]

[0127] 2. Detection of multimeric impurities in GLP-1 and GIP dual receptor agonist pharmaceutical compositions The compound of Formula I is a chemically synthesized polypeptide drug, and its formulations may aggregate to form multimers during production and storage. Multimers were detected by SEC.

[0128] Multimer detection was performed on Formulation 15, and the multimer content in the formulation under high temperature and light conditions was considered. The multimer limit was tentatively set at 2.0%. Specific data are shown in Table 12 and Figure 2.

[0129] [Table 12]

[0130] Surprisingly, the formulation sample showed no change in multimer content even after being stored for one month under the expected storage conditions (2-8°C), which is the storage condition expected to appear in the post-release instructions for this drug formulation. This indicates that the formulation designed in this disclosure solves the technical challenge of polypeptide drugs easily multimerizing and can ensure product stability after subsequent drug release. Furthermore, even after storage for 30 days under more stringent storage conditions, at 25°C and 30°C, the multimer content increased slightly, but remained within the limit. When stored at 2-8°C, the total irradiance was 1.2 x 10 6 lux·hr or more, and the near-ultraviolet lamp energy is 200W·hr / m 2 The above light irradiation increased the multimer content in the unpackaged sample, but it was still within the limit, and there was no significant change in the multimer content in the packaged sample.

[0131] In summary, the ingredients of this product were stable under appropriate storage conditions, and the polymer content was within the limit.

Claims

1. 1. A pharmaceutical composition comprising a GLP-1 and GIP dual receptor agonist, a stabilizer, and a buffer salt, wherein the GLP-1 and GIP dual receptor agonist is a compound of Formula I or a pharmaceutically acceptable salt, ester, solvate, optical isomer, tautomer, isotopic marker, or prodrug thereof. 【Chemical 1】

2. the pharmaceutical composition is a liquid formulation; The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition is preferably an injection.

3. 3. The pharmaceutical composition according to claim 2, wherein the concentration of the GLP-1 and GIP dual receptor agonist is 2 to 36 mg / mL.

4. The stabilizer is a polyol-based stabilizer, Preferably, the stabilizer is propylene glycol, mannitol or glycerol; More preferably, the pharmaceutical composition according to any one of claims 1 to 3, wherein the stabilizer is propylene glycol.

5. The pharmaceutical composition according to any one of claims 2 to 4, wherein the concentration of the stabilizer in the pharmaceutical composition is 10 to 20 mg / mL.

6. the buffer salt is selected from hydrogen phosphate, hydrogen phosphate hydrate, citrate or citrate hydrate; Preferably, the buffer salt is disodium hydrogen phosphate dodecahydrate or sodium citrate dihydrate; More preferably, the pharmaceutical composition according to any one of claims 1 to 5, wherein the buffer salt is disodium hydrogen phosphate dodecahydrate.

7. 7. The pharmaceutical composition according to claim 2, wherein the concentration of the buffer salt in the pharmaceutical composition is 7 to 23 mM.

8. the pH value of the pharmaceutical composition is 6.5 to 8.5; Preferably, the pH value of the pharmaceutical composition is 7.0 to 8.4; More preferably, the pH value of the pharmaceutical composition is 7.2 to 8.2; More preferably, the pharmaceutical composition according to any one of claims 1 to 7 has a pH value of 7.2 to 7.

6.

9. The pharmaceutical composition according to any one of claims 1 to 8, characterized in that it does not contain a bactericide.

10. A pharmaceutical composition comprising a GLP-1 and GIP dual receptor agonist of formula I, a stabilizer, and a buffer salt, wherein the stabilizer is propylene glycol and the buffer salt is disodium hydrogen phosphate dodecahydrate, the pH value of the pharmaceutical composition is 7.0 to 8.4, and the concentration of the GLP-1 and GIP dual receptor agonist of formula I is 5 to 30 mg / mL, the concentration of the propylene glycol is 10 to 20 mg / mL, and the concentration of the disodium hydrogen phosphate dodecahydrate is 7 to 23 mM, 【Chemistry 2】 The pharmaceutical composition is an injection.

11. 11. The pharmaceutical composition according to claim 10, wherein the pH value of the pharmaceutical composition is 7.2 to 8.2, and the concentration of disodium hydrogen phosphate dodecahydrate in the pharmaceutical composition is 10 to 20 mM.

12. 12. The pharmaceutical composition according to claim 10, wherein the pH of the pharmaceutical composition is 7.2 to 7.6, the concentration of the GLP-1 and GIP dual receptor agonist of formula I is 5 to 25 mg / mL, the concentration of the propylene glycol is 14 mg / mL, and the concentration of the disodium hydrogen phosphate dodecahydrate is 10 mM.

13. The pharmaceutical composition according to any one of claims 10 to 12, further comprising a pH adjuster and water, wherein the pH adjuster is hydrochloric acid and / or sodium hydroxide.

14. 14. A method for producing the pharmaceutical composition according to any one of claims 1 to 13, comprising the steps of: formulating a stabilizer, a buffer salt and a GLP-1 and GIP dual receptor agonist into a solution; and adjusting the pH value of the solution to a target value to obtain the pharmaceutical composition.

15. Use of the pharmaceutical composition according to any one of claims 1 to 13 in the manufacture of a medicament for preventing and / or treating a metabolic disorder-related disease, Preferably, the metabolic disorder-related disease is diabetes, diabetic complications, obesity, or obesity complications.

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