Use of FGF21 / GLP-1 fusion protein

Abstract

Provided is use of an FGF21 / GLP-1 fusion protein in the preparation of a drug for preventing and / or treating at least one of type 2 diabetes, obesity, steatohepatitis, and triglyceridemia. The FGF21 / GLP-1 fusion protein can effectively treat at least one of type 2 diabetes, obesity, steatohepatitis, and triglyceridemia. Especially, the fusion protein can reduce at least one of HbA1c, blood glucose, body weight, liver fat content, ALT, AST, glutamyl transpeptidase, triglyceride, total cholesterol, and low-density lipoprotein cholesterol in a subject, or can improve at least one of adiponectin, insulin sensitivity, and β cell function in the subject.

Description

Uses of FGF21 / GLP-1 fusion protein Technical Field

[0001] This application belongs to the field of biopharmaceuticals, specifically relating to the use of an FGF21 / GLP-1 fusion protein. Background Technology

[0002] Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease characterized by insulin resistance and insulin secretion defects. Traditional treatments for T2DM include lifestyle modifications, oral hypoglycemic agents, and insulin injections. However, these methods may not fully control blood sugar levels and may be accompanied by side effects.

[0003] In recent years, fusion protein therapy has attracted widespread attention as a novel treatment approach. Fusion proteins can enhance therapeutic effects by combining the properties of two or more bioactive molecules. FGF21 (fibroblast growth factor 21) and GLP-1 (glucagon-like peptide-1) are two proteins with the potential to regulate blood glucose and improve metabolic disorders. However, the efficacy and safety of fusion proteins are affected by various factors, such as the structure of the fusion protein, the buffer system, and the route of administration.

[0004] Therefore, further research is needed on the applications of the FGF21 / GLP-1 fusion protein in order to maximize therapeutic efficacy and minimize risks. Summary of the Invention

[0005] This application aims to at least partially address one of the technical problems existing in the prior art. To this end, this application provides a use of the FGF21 / GLP-1 fusion protein.

[0006] In a first aspect, this application proposes the use of the FGF21 / GLP-1 fusion protein in the preparation of a medicament for the prevention and / or treatment of at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia in individuals of need. The FGF21 / GLP-1 fusion protein of this application is effective in treating at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia, particularly by reducing at least one of HbA1c, blood glucose, body weight, liver fat content, ALT, AST, gamma-glutamyl transferase, and triglycerides in the user, or by increasing at least one of adiponectin, insulin sensitivity, and β-cell function in the user.

[0007] According to embodiments of this application, the drug is used to reduce at least one of the following indicators:

[0008] HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol.

[0009] According to embodiments of this application, the drug is used to improve at least one of the following indicators:

[0010] Adiponectin, insulin sensitivity, and β-cell function.

[0011] In a second aspect, this application proposes the use of the FGF21 / GLP-1 fusion protein in the prevention and / or treatment of diseases in individuals in need, said diseases being for the prevention and / or treatment of at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia. The FGF21 / GLP-1 fusion protein of this application is effective in treating at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia, particularly reducing at least one of HbA1c, blood glucose, weight, ALT, AST, triglycerides, liver fat content, total cholesterol, and LDL cholesterol in the user, or increasing at least one of adiponectin and β-cell function in the user.

[0012] According to embodiments of this application, the purpose is to reduce at least one of the following indicators:

[0013] HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, and triglycerides.

[0014] According to embodiments of this application, the purpose is to improve at least one of the following indicators:

[0015] Adiponectin, insulin sensitivity, and β-cell function.

[0016] In a third aspect, this application discloses an FGF21 / GLP-1 fusion protein for the prevention and / or treatment of at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia in individuals in need. The FGF21 / GLP-1 fusion protein of this application can effectively treat at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia in individuals in need, particularly by reducing at least one of HbA1c, blood glucose, weight, ALT, AST, liver fat content, triglycerides, total cholesterol, and low-density lipoprotein cholesterol in the user, or by increasing at least one of adiponectin and β-cell function in the user.

[0017] According to embodiments of this application, the FGF21 / GLP-1 fusion protein is used to reduce at least one of the following indicators:

[0018] HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol.

[0019] According to embodiments of this application, the FGF21 / GLP-1 fusion protein is used to improve at least one of the following indicators:

[0020] Adiponectin, insulin sensitivity, and β-cell function.

[0021] In a fourth aspect of this application, a method for preventing and / or treating a disease in an individual in need is proposed. According to embodiments of this application, the disease includes at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia; the method comprises administering a pharmaceutically acceptable dose of FGF21 / GLP-1 fusion protein to the individual in need.

[0022] According to embodiments of this application, the prevention and / or treatment are used to reduce at least one of the following indicators:

[0023] HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol.

[0024] According to embodiments of this application, the prevention and / or treatment are used to improve at least one of the following indicators:

[0025] Adiponectin, insulin sensitivity, and β-cell function.

[0026] According to embodiments of this application, the uses described in the first, second, and third aspects and the method described in the fourth method may further include at least one of the following technical features:

[0027] According to embodiments of this application, the FGF21 / GLP-1 fusion protein has the amino acid sequence shown in SEQ ID NO:1.

[0028] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 5.0 mg to 80.0 mg, for example, 5.0 mg, 5.1 mg, 6.0 mg, 7.0 mg, 8.0 mg, 9.0 mg, 10.0 mg, 10.2 mg, 11.0 mg, 12.0 mg, 13.0 mg, 14.0 mg, 15.0 mg, 15.3 mg, 16.0 mg, 17.0 mg, 18.0 mg, and 19.0 mg. 0mg, 20.0mg, 21.0mg, 22.0mg, 23.0mg, 24.0mg, 25.0mg, 25.5mg, 26.0mg, 27.0mg, 28.0mg, 29.0mg, 3 0.0mg, 30.6mg, 31.0mg, 32.0mg, 33.0mg, 34.0mg, 35.0mg, 36.0mg, 37.0mg, 37.5mg, 38.0mg, 39.0mg , 40.0mg, 41.0mg, 42.0mg, 43.0mg, 44.0mg, 45.0mg, 45.9mg, 46.0mg, 47.0mg, 47.6mg, 48.0mg, 49.0 mg, 50.0mg, 51.0mg, 52.0mg, 53.0mg, 54.0mg, 55.0mg, 56.0mg, 57.0mg, 58.0mg, 59.0mg, 60.0mg, 61 0.0 mg, 62.0 mg, 62.9 mg, 63.0 mg, 64.0 mg, 65.0 mg, 66.0 mg, 67.0 mg, 68.0 mg, 69.0 mg, 70.0 mg, 71.0 mg, 72.0 mg, 73.0 mg, 74.0 mg, 75.0 mg, 76.0 mg, 77.0 mg, 78.0 mg, 79.0 mg, 80.0 mg, or any two of these values ​​are used as the range between the endpoint values.

[0029] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 5.0 mg to 68.0 mg.

[0030] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 15.0 mg to 80.0 mg.

[0031] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 15.0 mg to 68.0 mg.

[0032] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 30.0 mg to 80.0 mg.

[0033] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 30.0 mg to 68.0 mg.

[0034] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 45.0 mg to 80.0 mg.

[0035] According to embodiments of this application, the effective dose of the FGF21 / GLP-1 fusion protein is 45.0 mg to 68.0 mg.

[0036] According to an embodiment of this application, the FGF21 / GLP-1 fusion protein is administered in the form of an injection.

[0037] According to embodiments of this application, the concentration of the FGF21 / GLP-1 fusion protein in the injection solution is 1–100 mg / mL, for example, 1.0 mg / mL, 2.0 mg / mL, 3.0 mg / mL, 4.0 mg / mL, 5.0 mg / mL, 6.0 mg / mL, 7.0 mg / mL, 8.0 mg / mL, 9.0 mg / mL, 10.0 mg / mL, 10.2 mg / mL, 11.0 mg / mL, 12.0 mg / mL, 13.0 mg / mL, 14.0 mg / mL, 15.0 mg / mL, 16.0 mg / mL, 17.0 mg / mL, 18.0 mg / mL, 19.0 mg / mL. 20.0mg / mL, 21.0mg / mL, 22.0mg / mL, 23.0mg / mL, 24.0mg / mL, 25.0mg / mL, 26.0mg / mL, 27.0mg / mL, 28.0mg / mL, 29.0mg / mL, 30.0mg / mL, 31.0mg / mL, 3 2.0mg / mL, 33.0mg / mL, 34.0mg / mL, 35.0mg / mL, 36.0mg / mL, 37.0mg / mL, 38.0mg / mL, 39.0mg / mL, 40.0mg / mL, 41.0mg / mL, 42.0mg / mL, 43.0mg / mL, 44. 0mg / mL, 45.0mg / mL, 46.0mg / mL, 47.0mg / mL, 48.0mg / mL, 49.0mg / mL, 50.0mg / mL, 51.0mg / mL, 52.0mg / mL, 53.0mg / mL, 54.0mg / mL, 55.0mg / mL, 56.0 mg / mL, 57.0mg / mL, 58.0mg / mL, 59.0mg / mL, 60.0mg / mL, 61.0mg / mL, 62.0mg / mL, 63.0mg / mL, 64.0mg / mL, 65.0mg / mL, 66.0mg / mL, 67.0mg / mL, 68.0m The values ​​can be g / mL, 69.0 mg / mL, 70.0 mg / mL, 71.0 mg / mL, 72.0 mg / mL, 73.0 mg / mL, 74.0 mg / mL, 75.0 mg / mL, 76.0 mg / mL, 77.0 mg / mL, 78.0 mg / mL, 79.0 mg / mL, 80.0 mg / mL, 85.0 mg / mL, 90.0 mg / mL, 95.0 mg / mL, 100.0 mg / mL, or any two of these values ​​as a range between endpoints; examples include 2–80 mg / mL, 5–70 mg / mL, 2–55 mg / mL, 10–65 mg / mL, or 20–60 mg / mL.

[0038] According to embodiments of this application, the injection solution further comprises at least one of a buffer solution, an osmotic pressure regulator, a surfactant, a preservative, and a metal chelating agent.

[0039] According to embodiments of this application, the buffer solution comprises at least one of histidine buffer, citrate buffer, phosphate buffer, and Tris buffer. Preferably, the buffer solution comprises histidine buffer. This invention has found that, in a histidine buffer system, the FGF21 / GLP-1 fusion protein of this invention exhibits good stability at multiple concentrations.

[0040] According to embodiments of this application, the concentration of the buffer solution in the injection solution is 5–30 mM, for example, 5.0 mM, 6.0 mM, 7.0 mM, 8.0 mM, 9.0 mM, 10.0 mM, 11.0 mM, 12.0 mM, 13.0 mM, 14.0 mM, 15.0 mM, 16.0 mM, 17.0 mM, 18.0 mM, 19.0 mM, 2… 0.0mM, 21.0mM, 22.0mM, 23.0mM, 24.0mM, 25.0mM, 26.0mM, 27.0mM, 28.0mM, 29.0mM, 30.0mM, or any two of these values ​​can be used as a range between endpoint values; examples include 5–25mM, 8–25mM, 5–20mM, 8–12mM, or 10–20mM.

[0041] According to embodiments of this application, the osmotic pressure regulator includes at least one of mannitol, sucrose, trehalose, and sodium chloride.

[0042] According to embodiments of this application, the concentration of the osmotic pressure regulator in the injection solution is 30–110 mg / mL, for example, 30.0 mg / mL, 31.0 mg / mL, 32.0 mg / mL, 33.0 mg / mL, 34.0 mg / mL, 35.0 mg / mL, 36.0 mg / mL, 37.0 mg / mL, 38.0 mg / mL, 39.0 mg / mL, 40.0 mg / mL, and 41.0 mg / mL. / mL, 42.0mg / mL, 43.0mg / mL, 44.0mg / mL, 45.0mg / mL, 46.0mg / mL, 47.0mg / mL, 48.0mg / mL, 49.0mg / mL, 50.0mg / mL, 51.0mg / mL, 52.0mg / mL, 53.0mg / mL, 54.0mg / mL, 55.0mg / mL, 56.0mg / mL, 57.0mg / mL g / mL, 58.0mg / mL, 59.0mg / mL, 60.0mg / mL, 61.0mg / mL, 62.0mg / mL, 63.0mg / mL, 64.0mg / mL, 65.0 mg / mL, 66.0mg / mL, 67.0mg / mL, 68.0mg / mL, 69.0mg / mL, 70.0mg / mL, 71.0mg / mL, 72.0mg / mL, 73.0 mg / mL, 74.0 mg / mL, 75.0 mg / mL, 76.0 mg / mL, 77.0 mg / mL, 78.0 mg / mL, 79.0 mg / mL, 80.0 mg / mL, 85.0 mg / mL, 90.0 mg / mL, 95.0 mg / mL, 100.0 mg / mL, 105.0 mg / mL, 110.0 mg / mL, or any two of these values ​​are used as the range between the endpoint values.

[0043] According to an embodiment of this application, the concentration of the osmotic pressure regulator in the injection solution is 1 to 10 mg / mL, for example, 1.0 mg / mL, 2.0 mg / mL, 3.0 mg / mL, 4.0 mg / mL, 5.0 mg / mL, 6.0 mg / mL, 7.0 mg / mL, 8.0 mg / mL, 9.0 mg / mL, 10.0 mg / mL, or any two of these values ​​as a range between endpoints.

[0044] According to embodiments of this application, the surfactant includes at least one of polysorbate 80, polysorbate 20, and poloxamer 188.

[0045] According to embodiments of this application, the concentration of the surfactant in the injection solution is 0.1–10 mg / mL, for example, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 1.1 mg / mL, 1.2 mg / mL, 1.3 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 3.0 mg / mL, 4.0 mg / mL, 5.0 mg / mL, 6.0 mg / mL, 7.0 mg / mL, 8.0 mg / mL, 9.0 mg / mL, 10.0 mg / mL, or any two of these values ​​as endpoints; exemplary values ​​are 0.1–1.5 mg / mL, 6–10 mg / mL, or 0.1–0.7 mg / mL.

[0046] According to embodiments of this application, the preservative is at least one of phenol and m-cresol.

[0047] According to embodiments of this application, the concentration of the preservative in the injection solution is 0.1–10 mg / mL, for example, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 2.0 mg / mL, 3.0 mg / mL, 4.0 mg / mL, 5.0 mg / mL, 6.0 mg / mL, 7.0 mg / mL, 8.0 mg / mL, 9.0 mg / mL, 10.0 mg / mL, or any two of these values ​​as endpoints; an exemplary value is 1.0–5.0 mg / mL.

[0048] According to embodiments of this application, the metal chelating agent is at least one of EDTA, sodium calcium edetate, disodium edetate, and sodium calcium edetate.

[0049] According to embodiments of this application, the concentration of the metal chelating agent in the injection solution is 0.01–1.0 mg / mL, for example, 0.01 mg / mL, 0.02 mg / mL, 0.03 mg / mL, 0.04 mg / mL, 0.05 mg / mL, 0.06 mg / mL, 0.07 mg / mL, 0.08 mg / mL, 0.09 mg / mL, 1.0 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, or any two of these values ​​as endpoints; an exemplary range is 0.05–0.5 mg / mL or 0.01–1.0 mg / mL.

[0050] According to embodiments of this application, the buffer solution is a histidine buffer, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 10–100 mg / mL, for example, 10.0 mg / mL, 11.0 mg / mL, 12.0 mg / mL, 13.0 mg / mL, 14.0 mg / mL, 15.0 mg / mL, 16.0 mg / mL, 17.0 mg / mL, 18.0 mg / mL, 19.0 mg / mL, 20.0 mg / mL, 21.0 mg / mL, 22.0 mg / mL, 23.0 mg / mL, 24.0 mg / mL, 2 5.0mg / mL, 26.0mg / mL, 27.0mg / mL, 28.0mg / mL, 29.0mg / mL, 30.0mg / mL, 31.0mg / mL, 32.0mg / mL, 33.0mg / mL, 34.0mg / mL, 35.0mg / mL, 36.0mg / mL, 37.0mg / mL, 38.0mg / mL, 39.0mg / mL, 40.0mg / mL, 41.0mg / mL, 42.0mg / mL, 43.0mg / mL, 44.0mg / mL, 45.0mg / mL, 46.0mg / mL, 47.0mg / mL, 48.0mg / mL, 49.0mg / mL, 50.0mg / mL, 51.0mg / mL, 52.0mg / mL, 53.0mg / mL, 54.0mg / mL, 55.0mg / mL, 56.0mg / mL, 57.0mg / mL , 58.0mg / mL, 59.0mg / mL, 60.0mg / mL, 61.0mg / mL, 62.0mg / mL, 63.0mg / mL, 64.0mg / mL, 65.0mg / mL, 66.0mg / mL, 67.0mg / mL, 68.0mg / mL 69.0 mg / mL, 70.0 mg / mL, 71.0 mg / mL, 72.0 mg / mL, 73.0 mg / mL, 74.0 mg / mL, 75.0 mg / mL, 76.0 mg / mL, 77.0 mg / mL, 78.0 mg / mL, 79.0 mg / mL, 80.0 mg / mL, 85.0 mg / mL, 90.0 mg / mL, 95.0 mg / mL, 100.0 mg / mL, or any two of these values ​​are used as a range between endpoints; examples include 25–95 mg / mL, 25–90 mg / mL, and 25–85 mg / mL.

[0051] According to embodiments of this application, the buffer solution is a histidine buffer, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25–80 mg / mL, for example, 25.0 mg / mL, 26.0 mg / mL, 27.0 mg / mL, 28.0 mg / mL, 29.0 mg / mL, 30.0 mg / mL, 31.0 mg / mL, 32.0 mg / mL, 33.0 mg / mL, 34.0 mg / mL. L, 35.0mg / mL, 36.0mg / mL, 37.0mg / mL, 38.0mg / mL, 39.0mg / mL, 40.0mg / mL, 41.0mg / mL, 42.0mg / mL, 4 3.0mg / mL, 44.0mg / mL, 45.0mg / mL, 46.0mg / mL, 47.0mg / mL, 48.0mg / mL, 49.0mg / mL, 50.0mg / mL, 51.0 mg / mL, 52.0mg / mL, 53.0mg / mL, 54.0mg / mL, 55.0mg / mL, 56.0mg / mL, 57.0mg / mL, 58.0mg / mL, 59.0mg / mL, 60.0mg / mL, 61.0mg / mL, 62.0mg / mL, 63.0mg / mL, 64.0mg / mL, 65.0mg / mL, 66.0mg / mL, 67.0mg / mL 68.0 mg / mL, 69.0 mg / mL, 70.0 mg / mL, 71.0 mg / mL, 72.0 mg / mL, 73.0 mg / mL, 74.0 mg / mL, 75.0 mg / mL, 76.0 mg / mL, 77.0 mg / mL, 78.0 mg / mL, 79.0 mg / mL, 80.0 mg / mL, or any two of these values ​​are used as the range between the endpoints; for example, 25–55 mg / mL.

[0052] According to embodiments of this application, the concentration of the histidine buffer in the injection solution is 5–30 mM, for example, 5.0 mM, 6.0 mM, 7.0 mM, 8.0 mM, 9.0 mM, 10.0 mM, 11.0 mM, 12.0 mM, 13.0 mM, 14.0 mM, 15.0 mM, 16.0 mM, 17.0 mM, 18.0 mM, or 19.0 mM. 20.0mM, 21.0mM, 22.0mM, 23.0mM, 24.0mM, 25.0mM, 26.0mM, 27.0mM, 28.0mM, 29.0mM, 30.0mM, or any two of these values ​​can be used as the range between endpoint values; examples include 5–25mM, 8–25mM, 8–20mM, 10–20mM, and 8–12mM.

[0053] According to an embodiment of this application, the pH value of the injection solution is 6.7 to 7.5, preferably 6.9 to 7.5.

[0054] According to embodiments of this application, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the injection solution is 30–50 mg / mL; or

[0055] The osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the injection solution is 60–100 mg / mL, preferably 80–100 mg / mL; or

[0056] The osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the injection solution is 40-100 mg / mL, preferably 40-70 mg / mL, and more preferably 50-70 mg / mL.

[0057] According to embodiments of this application, the surfactant is polysorbate 80, and the concentration of the surfactant in the injection solution is 0.1–2 mg / mL, preferably 0.1–1.5 mg / mL, and more preferably 0.4–1.3 mg / mL; or

[0058] The surfactant is polysorbate 20, and the concentration of the surfactant in the injection solution is 0.1-1.5 mg / mL, preferably 0.1-0.5 mg / mL.

[0059] According to an embodiment of this application, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 2-25 mg / mL, and the buffer solution is citrate buffer.

[0060] According to embodiments of this application, the concentration of the citrate buffer in the injection solution is 5-30 mM, preferably 5-25 mM, 8-25 mM, or 8-20 mM, and more preferably 10-20 mM or 8-12 mM.

[0061] According to an embodiment of this application, the pH value of the injection solution is 6.4 to 8.0.

[0062] According to embodiments of this application, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the injection solution is 30–50 mg / mL; or

[0063] The osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the injection solution is 60–100 mg / mL, preferably 80–100 mg / mL; or

[0064] The osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the injection solution is 40–100 mg / mL, preferably 40–70 mg / mL, more preferably 50–70 mg / mL; or

[0065] The osmotic pressure regulator is sodium chloride, and the concentration of the osmotic pressure regulator in the injection solution is 1-10 mg / mL, preferably 3-9 mg / mL.

[0066] According to embodiments of this application, the surfactant includes at least one of polysorbate 80, polysorbate 20, and poloxamer 188, preferably polysorbate 80.

[0067] According to an embodiment of this application, the concentration of the surfactant in the injection solution is 0.1–10 mg / mL, preferably 0.1–1.5 mg / mL, 0.1–1.0 mg / mL, or 6–10 mg / mL.

[0068] According to embodiments of this application, the surfactant is poloxamer 188, and the concentration of the surfactant in the injection solution is 0.2–10 mg / mL, preferably 0.2–1.5 mg / mL; or

[0069] The surfactant is polysorbate 80 or polysorbate 20, and the concentration of the surfactant in the injection solution is 0.1 to 1.5 mg / mL, preferably 0.1 to 0.8 mg / mL.

[0070] According to embodiments of this application, the surfactant is selected from polysorbate 80, and the metal chelating agent is selected from at least one of EDTA, calcium sodium edetate, and disodium edetate.

[0071] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and polysorbate 80, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3–25 mg / mL; the concentration of the citrate buffer in the injection solution is 5–20 mM; the concentration of the mannitol in the injection solution is 40–50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1–1.5 mg / mL; the concentration of at least one of EDTA, calcium sodium edetate, and disodium edetate in the injection solution is 0.01–1.0 mg / mL; and the pH value of the injection solution is 6.4–8.0.

[0072] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and polysorbate 80, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3–25 mg / mL; the concentration of the citrate buffer in the injection solution is 8–20 mM; the concentration of the mannitol in the injection solution is 40–50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1–1.0 mg / mL; the concentration of at least one of EDTA, calcium sodium edetate, and disodium edetate in the injection solution is 0.01–1.0 mg / mL; and the pH value of the injection solution is 6.4–8.0.

[0073] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and polysorbate 80, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3–25 mg / mL; the concentration of the citrate buffer in the injection solution is 8–20 mM; the concentration of the mannitol in the injection solution is 40–50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1–0.5 mg / mL; the concentration of at least one of EDTA, calcium sodium edetate, and disodium edetate in the injection solution is 0.01–1.0 mg / mL; and the pH value of the injection solution is 6.4–8.0.

[0074] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and polysorbate 80, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 10-100 mg / mL, preferably 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, 25-80 mg / mL, 25-75 mg / mL, 25-70 mg / mL, 25-65 mg / mL, 25-60 mg / mL, 25-55 mg / mL, or 25-50 mg / mL; the histidine... The concentration of the acid buffer in the injection solution is 8–25 mM, preferably 10–20 mM; the concentration of the mannitol in the injection solution is 40–50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.4–1.5 mg / mL or 0.4–1.3 mg / mL; the concentration of at least one of EDTA, calcium sodium edetate, and disodium edetate in the injection solution is 0.01–1.0 mg / mL; and the pH value of the injection solution is 6.9–7.5.

[0075] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and polysorbate 80, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 40–90 mg / mL; the concentration of the histidine buffer in the injection solution is 10–20 mM; the concentration of the mannitol in the injection solution is 40–50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.4–1.3 mg / mL; the concentration of at least one of EDTA, calcium sodium edetate, and disodium edetate in the injection solution is 0.01–1.0 mg / mL; and the pH value of the injection solution is 6.9–7.5.

[0076] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol and poloxamer 188, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises as shown in SEQ ID. The amino acid sequence shown in NO:1 indicates that the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 10–100 mg / mL, preferably 25–95 mg / mL, 25–90 mg / mL, 25–85 mg / mL, 25–80 mg / mL, 25–75 mg / mL, 25–70 mg / mL, 25–65 mg / mL, 25–60 mg / mL, 25–55 mg / mL, or 25–50 mg / mL; the concentration of the histidine buffer in the injection solution is 8–25 mM, preferably 10–20 mM; the concentration of the mannitol in the injection solution is 40–50 mg / mL; the concentration of the poloxamer 188 in the injection solution is 0.2–10 mg / mL, preferably 0.2–1.5 mg / mL; and the pH value of the injection solution is 6.9–7.5.

[0077] In an optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and polysorbate 20, and optionally at least one of EDTA, sodium calcium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises as shown in SEQ ID. The amino acid sequence shown in NO:1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 10-100 mg / mL, preferably 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, 25-80 mg / mL, 25-75 mg / mL, 25-70 mg / mL, 25-65 mg / mL, 25-60 mg / mL, 25-55 mg / mL, or 25-50 mg / mL; the concentration of the histidine buffer in the injection solution is 8-25 mM, preferably 10-20 mM; the concentration of the mannitol in the injection solution is 40-50 mg / mL; the concentration of the polysorbate 20 in the injection solution is 0.1-1.5 mg / mL, preferably 0.1-0.8 mg / mL; and the pH value of the injection solution is 6.9-7.5.

[0078] In one optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose or sodium chloride, and polysorbate 80, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 10-100 mg / mL, preferably 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, 25-80 mg / mL, 25-75 mg / mL, 25-70 mg / mL, 25-65 mg / mL, 25-60 mg / mL, 25-55 mg / mL, or 25-50 mg / mL; The histidine buffer solution has a concentration of 8–25 mM in the injection solution, preferably 10–20 mM; the sucrose concentration in the composition is 60–100 mg / mL, preferably 80–100 mg / mL; the trehalose concentration in the composition is 40–100 mg / mL, preferably 40–70 mg / mL, more preferably 50–70 mg / mL; the sodium chloride concentration in the composition is 1–10 mg / mL, preferably 3–9 mg / mL. The polysorbate 80 concentration in the injection solution is 0.4–1.5 mg / mL; the pH of the injection solution is 6.9–7.5.

[0079] In one optional embodiment of this application, the injection solution comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose or sodium chloride, and poloxamer 188, and optionally at least one of EDTA, calcium sodium edetate, and disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO:1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 10-100 mg / mL, preferably 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, 25-80 mg / mL, 25-75 mg / mL, 25-70 mg / mL, 25-65 mg / mL, 25-60 mg / mL, 25-55 mg / mL, or 25-50 mg / mL; The histidine buffer solution has a concentration of 8–25 mM in the injection solution, preferably 10–20 mM; the sucrose concentration in the composition is 60–100 mg / mL, preferably 80–100 mg / mL; the trehalose concentration in the composition is 40–100 mg / mL, preferably 40–70 mg / mL, more preferably 50–70 mg / mL; the sodium chloride concentration in the composition is 1–10 mg / mL, preferably 3–9 mg / mL. The poloxamer 188 concentration in the injection solution is 0.2–10 mg / mL, preferably 0.2–1.5 mg / mL; the pH of the injection solution is 6.9–7.5.

[0080] The aforementioned injection solution of the present invention can effectively control the generation of visible foreign matter and aggregates, and can improve the physical and chemical stability of fusion proteins, thereby reducing potential safety risks and extending shelf life.

[0081] According to embodiments of this application, the FGF21 / GLP-1 fusion protein is administered via fixed-dose administration or titration administration.

[0082] According to an embodiment of this application, the FGF21 / GLP-1 fusion protein is administered via titration.

[0083] According to the embodiments of this application, the FGF21 / GLP-1 fusion protein is administered once a month, once every four weeks, once every three weeks, once every two weeks, once a week, twice a week, three times a week, or once a day. Beneficial effects:

[0084] 1. The FGF21 / GLP-1 fusion protein of this application has at least one of the following uses:

[0085] HbA1c decreased by approximately 0.40% or more;

[0086] Fasting blood glucose decreased by approximately 1.700 mmol / L or more;

[0087] MRI-PDFF decreased by approximately 30%;

[0088] ALT decreased by approximately 30%;

[0089] AST decreased by approximately 25%.

[0090] 2. Compared with the control drug, the FGF21 / GLP-1 fusion protein of this application has better effects in lowering blood sugar, lowering blood lipids and reducing body weight.

[0091] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0092] The above and / or additional aspects and advantages of this application will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0093] Figure 1 is a semi-logarithmic graph of mean blood drug concentration-time for FGF21 / GLP-1, GLP-1 and FGF21 in T2DM subjects at each dosing stage in Example 2 of this application;

[0094] Figure 2 shows the changes in HbA1c and fasting blood glucose of T2DM subjects in Example 2 of this application compared with baseline.

[0095] Figure 3 shows the changes in blood glucose, insulin levels, insulin sensitivity, and β-cell function of T2DM subjects in Example 2 of this application compared to baseline during a standard meal test.

[0096] Figure 4 shows the changes in adiponectin levels from baseline in T2DM subjects in Example 2 of this application.

[0097] Figure 5 shows the changes in triglycerides from baseline in T2DM subjects in Example 2 of this application;

[0098] Figure 6 shows the MRI-PDFF and fasting weight changes from baseline of the T2DM subjects in Example 2 of this application;

[0099] Figure 7 shows the changes in liver enzymes in T2DM subjects compared to baseline in Example 2 of this application;

[0100] Figure 8 shows the correlation results between the primary efficacy endpoints and drug exposure levels in T2DM subjects in Example 2 of this application;

[0101] Figure 9 shows the blood glucose AUC in Example 1 of this application. 0-2h AUC 0-4hThe result of the change compared to the baseline. Detailed Implementation

[0102] The embodiments of this application are described in detail below. The embodiments described below are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0103] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this application, unless otherwise stated, "multiple" means two or more.

[0104] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0105] In this article, the terms "type 2 diabetes," "T2D," or "T2DM" refer to a metabolic disease that causes glucose to accumulate in the bloodstream and is characterized by high blood glucose levels caused by a lack of insulin or the body's inability to use insulin effectively. Blood glucose levels are measured in milligrams per deciliter (mg / dL) or mmol / L.

[0106] In this article, the term "obesity" refers to an abnormal or excessive accumulation of fat that poses a health risk. A rough population measure of obesity is the Body Mass Index (BMI = weight (kg) / height (m)). 2 A BMI of 30 or higher is calculated by dividing a person's weight (in kilograms) by the square of their height (in meters). People with a BMI of 30 or higher are generally considered obese.

[0107] In this article, the term "steatohepatitis" refers to a disease caused by excessive fat accumulation in the liver, leading to an inflammatory response in the liver. It is also known as fatty liver disease, including but not limited to non-alcoholic steatohepatitis and simple fatty liver.

[0108] In this article, the term "triglyceridemia" refers to high blood levels of triglycerides (congestion). Elevated triglyceride levels are associated with atherosclerosis and predispose to cardiovascular disease (e.g., contributing to an increased risk of cardiovascular disease) even in the absence of hypercholesterolemia (high cholesterol levels).

[0109] In this document, the terms “comprising” or “including” are open-ended expressions, meaning that they include the contents specified in this application but do not exclude other contents.

[0110] In this document, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition that may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0111] In this document, the term "treatment" refers to the administration of a drug or compound to an individual to achieve a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of a disease or its symptoms, and / or therapeutic in terms of partial or complete cure of a disease and / or adverse effects caused by the disease. As used herein, "treatment" encompasses diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of a condition in an individual who is susceptible but has not yet been diagnosed with the disease; (b) inhibition of disease, such as blocking disease progression; or (c) relief of disease, such as reducing symptoms associated with the disease. As used herein, "treatment" encompasses any administration of a drug or compound to an individual to treat, cure, relieve, improve, reduce, or inhibit the individual's disease, including but not limited to administration of a drug containing a compound described herein to an individual in need.

[0112] As used herein, the term “application” means administration by a physician, nurse, healthcare provider, patient, or any other individual, including self-application. This includes not only delivery into the body but also prescription, distribution, or any other means of facilitating delivery.

[0113] In this article, the term "patient" refers to a mammal that requires treatment for its symptoms or condition.

[0114] In this document, the term "individual in need" means a mammal, such as a human, suffering from a condition, disease, ailment, or symptom (e.g., including those listed herein) that requires treatment or therapy. Humans are the preferred individual to be treated.

[0115] In this document, the term "titration dose" refers to a dose smaller than the highest effective dose required by the patient. As used herein, this application covers a "titration dose" that can be converted to the highest effective dose required, or, if observed to be an effective dose required by the patient, can be converted to a "maintenance dose" and can be administered for an extended period of more than four weeks.

[0116] In this document, the term "maintenance dose" refers to the dose that is the highest effective dose required by the patient, and when the maintenance dose is less than the required highest effective dose, this maintenance dose may become a titration dose. That is, if for a specific patient, the "15.3 mg" maintenance dose covered by this application is not the required highest effective dose, then the 15.3 mg maintenance dose will, in turn, become a titration dose because the dose for that specific patient will be increased until the next highest maintenance dose covered by this application is achieved, for example, see the dosing of group 3 or group 4 subjects in this application for T2D.

[0117] In this paper, the terms "FGF21 / GLP-1 fusion protein" and "GLP-1-Fc-FGF21 fusion protein" refer to fusion proteins composed of FGF21, Fc domain, GLP-1 and linkers, wherein FGF21, Fc domain and GLP-1 are linked by linkers and have the amino acid sequence shown in SEQ ID NO:1.

[0118] In this document, the term "HbA1c" refers to glycated hemoglobin level, which is formed when hemoglobin binds to glucose in the blood. HbA1c level is a commonly used indicator of glycemic control in diabetic patients, and a decreased HbA1c level generally indicates improved glycemic control. In the case of the method of this application, the method of this application results in a decrease in HbA1c. In some embodiments, a decrease in HbA1c refers to a reduction in HbA1c level relative to treatment with a lower dose of FGF21 / GLP-1 fusion protein.

[0119] In this article, the term “MRI-PDFF”, also known as magnetic resonance proton density fat fraction, is a non-invasive MRI imaging technique that can accurately assess the accumulation of triglycerides in the liver and has high accuracy and good reproducibility in the diagnosis of non-alcoholic fatty liver disease (NAFLD).

[0120] In this article, the term "AST" stands for alanine aminotransferase; "ALT" stands for aspartate aminotransferase.

[0121] The following will explain the solution of this application with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of this application. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0122] Preparation Example 1: Preparation of FGF21 / GLP-1 fusion protein injection solution

[0123] An exemplary formulation of the FGF21 / GLP-1 fusion protein injection is as follows:

[0124] The protein consists of citric acid monohydrate (0.14 mg / ml) + sodium citrate dihydrate (2.74 mg / ml), totaling 10 mM; histidine (1.552 mg / ml), totaling 10 mM; and the amino acid sequence of the FGF21 / GLP-1 fusion protein (hereinafter referred to as FGF21 / GLP-1) is shown in SEQ ID NO:1.

[0125] The preparation method of the injection solution is as follows: Weigh the excipients and FGF21 / GLP-1 fusion protein according to the prescription in the table above, mix them, and if necessary, adjust the pH of the preparation to the appropriate value using 2% hydrochloric acid solution or 2% sodium hydroxide solution. Finally, add water for injection to make up the volume. After the solution is prepared, filter the solution through a series of 0.45μm and 0.22μm PVDF filter membranes into 1ml thin pre-filled syringes, filling each syringe with 1ml, and seal with a rubber stopper.

[0126] Furthermore, the hypoglycemic, lipid-lowering, and weight-reducing effects of prescriptions 1 to 7 were studied using the db / db mouse model. The results showed that prescriptions 1 to 7 could all achieve hypoglycemic, lipid-lowering, and weight-reducing effects.

[0127] Example 1: Study on the therapeutic effect in obese subjects

[0128] Eight obese subjects were given a single dose of 5.1 mg of the FGF21 / GLP-1 fusion protein injection prepared according to Preparation Example 1, and two subjects received a placebo (containing only the absence of the FGF21 / GLP-1 fusion protein compared to the FGF21 / GLP-1 fusion protein injection), for a total of 10 subjects. The safety, tolerability, pharmacokinetics, and pharmacodynamics of the FGF21 / GLP-1 fusion protein injection were investigated.

[0129] Obese participants must meet all of the following criteria to be eligible:

[0130] 1) Sign an informed consent form before the trial and fully understand the trial content, process, and possible adverse reactions;

[0131] 2) Able to complete the test according to the requirements of the test plan;

[0132] 3) Subjects (including their partners) voluntarily use effective contraception within 6 months from screening to the last time they receive the investigational drug;

[0133] 4) When signing informed consent, the applicant must be 18 years of age or younger and 45 years of age or older.

[0134] 5) Males weighing ≥50kg and females weighing ≥45kg. For obese subjects, 28≤BMI≤45kg / m2;

[0135] 6) Normal or abnormal physical examination and vital signs have no clinical significance.

[0136] In addition, subjects who meet any of the following exclusion criteria are not eligible to participate in this study:

[0137] 1) Individuals who smoked an average of more than 5 cigarettes per day in the 3 months prior to the experiment;

[0138] 2) Individuals with allergies (allergies to multiple medications and foods);

[0139] 3) History of alcohol abuse (consuming 14 units of alcohol per week: 1 unit = 285 mL of beer, or 25 mL of spirits, or 100 mL of wine);

[0140] 4) Individuals with a history of drug abuse or drug use within the five years prior to screening;

[0141] 5) Donating blood (>300mL) or experiencing significant blood loss (>400mL) within 3 months prior to screening;

[0142] 6) Having any condition that increases the risk of bleeding, such as hemorrhoids, acute gastritis, or gastric and duodenal ulcers;

[0143] 7) Individuals with a personal or family history of medullary thyroid carcinoma (MTC) or multiple endocrine neoplasia type 2 (MEN2), or those with hereditary diseases that predispose them to medullary thyroid carcinoma;

[0144] 8) Individuals who have received a live attenuated vaccine or a COVID-19 vaccine within one month prior to screening, or who plan to receive a vaccine during the trial period;

[0145] 9) Have taken any prescription drugs, over-the-counter drugs, any vitamin products, or traditional Chinese medicine within 14 days prior to screening;

[0146] 10) Those who have recently made significant changes to their diet or exercise habits;

[0147] 11) Subjects who have received the investigational drug or participated in a clinical trial within the three months prior to screening (subjects who withdraw from the study before treatment, i.e., who are not randomized or do not receive treatment, may be enrolled in this study);

[0148] 12) Abnormalities on electrocardiograms are clinically significant;

[0149] 13) Women who are breastfeeding or pregnant, or women of childbearing age with a positive pregnancy test;

[0150] 14) Clinically significant abnormalities in clinical laboratory tests, or other clinical findings within the 12 months prior to screening showing clinical significance of the following diseases (including but not limited to gastrointestinal, kidney, liver, nervous, blood, endocrine, tumor, lung, immune, mental or cardiovascular diseases);

[0151] 15) Positive screening results for hepatitis B surface antigen, hepatitis C antibody or core antigen, HIV antibody, and Treponema pallidum antibody [For those with positive Treponema pallidum antibody, a rapid plasma reagin (RPR) test is required. If the RPR is also positive, the individual should be excluded].

[0152] 16) An acute illness or concomitant medication occurred from the screening stage until the first administration of the investigational drug;

[0153] 17) Individuals who test positive for alcohol screening or urine drug screening;

[0154] 18) Subjects who, according to the researchers, have other factors that make them unsuitable to participate in this trial.

[0155] 1. Pharmacokinetics:

[0156] Serum concentrations of FGF21 / GLP-1, GLP-1 (intact GLP-1 + Fc), and FGF21 (Fc + intact FGF21) will be quantitatively measured separately. Key pharmacokinetic parameters include: peak concentration (C0). max Area under the plasma concentration-time curve (AUC) 0-t AUC 0-∞ Peak time (T) max ), elimination phase half-life (t) 1 / 2 ), mean residence time (MRT), clearance rate (CL / F), apparent distribution volume (Vz / F), elimination rate constant (λz), percentage of AUC0-∞ extrapolated (%AUCex), etc.

[0157] 2. Pharmacodynamics:

[0158] 1) Oral Glucose Trial (OGTT): 300 mL of 25% glucose solution was orally ingested on the mornings of days 1, 3, and 7. Timing began from the first sip of the glucose solution. Blood samples were collected within 0.5 hours before ingestion and at 0.5, 1, 2, 3, and 4 hours after ingestion to measure blood glucose concentration. Fasting blood glucose on day 3 could be collected during blood biochemistry testing; no additional blood sampling was required.

[0159] 2) Fasting blood glucose: During the screening period, D-1, D2, D4, D5, D8, D10, D15 or early withdrawal from the visit. Among them, the fasting blood glucose on D-1 can be collected during the OGTT test. During the screening period, D10, D15 or early withdrawal from the visit, the fasting blood glucose can be collected during the blood biochemistry test, without the need for additional blood collection.

[0160] 3) Adiponectin: D-1, D3, D10, D15 or early withdrawal from the visit.

[0161] 4) Fasting blood lipids [triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), TC / HDL-C ratio, apolipoprotein B (ApoB)]: During the screening period, D-1, D3, D10, D15, or early withdrawal from the visit, except for D-1, fasting blood lipids at other time points can be collected during blood biochemistry tests, without the need for additional blood collection.

[0162] 3. Safety:

[0163] The safety and tolerability of FGF21 / GLP-1 fusion protein injection were evaluated in subjects via subcutaneous abdominal injection. Safety evaluation primarily included adverse events, laboratory tests (complete blood count, blood biochemistry, coagulation function, thyroid function, urinalysis), vital signs, physical examination, and 12-lead electrocardiogram.

[0164] The test results are as follows:

[0165] 1. As shown in Figure 9, blood glucose AUC was observed in obese subjects on both days 3 and 7. 0-2h AUC 0-4h It decreased compared to the baseline.

[0166] 2. The FGF21 / GLP-1 fusion protein injection showed good safety and tolerability in obese subjects after a single dose.

[0167] Example 2: Study on the therapeutic effect in subjects with type 2 diabetes

[0168] The FGF21 / GLP-1 fusion protein injection prepared according to Preparation Example 1 was administered multiple times to subjects with type 2 diabetes mellitus (T2DM). Five dose groups were established (as shown in Table 1), namely groups 1 through 5, with tentatively set doses of 5.1 mg, 15.3 mg, 30.6 mg, 45.9 mg, and 68.0 mg, respectively. Groups 1 and 2 were fixed-dose groups, while groups 3 through 5 were titrated-dose groups, with titration protocols shown in Table 1. Each group enrolled 12 T2DM subjects, of whom 10 received the FGF21 / GLP-1 injection and 2 received a placebo (which, compared to the FGF21 / GLP-1 fusion protein injection, only lacked the FGF21 / GLP-1 fusion protein). Administration was once weekly for 5 weeks, administered via subcutaneous abdominal injection after meals on days 1, 8, 15, 22, and 29. The trial was conducted using a dose-escalation approach. After the tolerance assessment of D15 was completed in the previous dose group, if it was tolerable, the dose could be escalated to the next dose group.

[0169] Table 1 Note: T2DM refers to subjects with type 2 diabetes.

[0170] Among them, subjects with type 2 diabetes (T2DM) must meet all of the following requirements:

[0171] 1) Age 18 ≤ age ≤ 65, gender not limited;

[0172] 2) 24 kg / m² ≤ Body Mass Index (BMI) ≤ 35 kg / m² 2 ;

[0173] 3) Individuals diagnosed with type 2 diabetes according to the 2020 edition of the Chinese Guidelines for the Prevention and Treatment of Type 2 Diabetes, who are newly diagnosed at the time of screening, or whose blood glucose control is still poor after receiving only diet and exercise therapy within 3 months prior to screening, or who have received stable doses of metformin for ≥3 months prior to screening.

[0174] 4) At the screening stage, 7.0% ≤ glycated hemoglobin (HbA1c) ≤ 10.5% and fasting plasma glucose (FPG) ≤ 13.9 mmol / L;

[0175] 5) Fibroscan score ≤12.5kPa during the screening period, and abdominal ultrasound examination during the screening period shows fatty liver but no signs of cirrhosis.

[0176] In addition, subjects who meet any of the following exclusion criteria are not eligible to participate in this study:

[0177] 1) Individuals with allergies (allergies to multiple medications and foods);

[0178] 2) History of alcohol abuse (consuming 14 units of alcohol per week: 1 unit = 285 mL of beer, or 25 mL of spirits, or 100 mL of wine);

[0179] 3) Individuals with a history of drug abuse or drug use within the five years prior to screening;

[0180] 4) Donating blood (>300mL) or experiencing significant blood loss (>400mL) within 3 months prior to screening;

[0181] 5) Subjects with type 1 diabetes;

[0182] 6) Those clinically diagnosed with cirrhosis;

[0183] 7) Screening for individuals who have experienced acute complications of diabetes (diabetic ketoacidosis, nonketotic hyperosmolar coma, diabetic lactic acidosis, etc.) within the past 6 months;

[0184] 8) Individuals suffering from proliferative retinopathy and / or macular degeneration requiring urgent treatment;

[0185] 9) Having experienced one or more severe hypoglycemic episodes or a history of recurrent hypoglycemia within the 6 months prior to screening (more than 3 episodes of hypoglycemia within the 3 months).

[0186] 10) Those who have a history of acute or chronic pancreatitis before randomization, or have acute or chronic pancreatitis at the time of randomization, or have serum amylase or lipase ≥2×Upper Limit of Normal (ULN) at the time of screening, or have undergone pancreatectomy.

[0187] 11) Known clinically significant gastric emptying abnormalities (such as severe diabetic gastroparesis or gastric outlet obstruction), or having undergone gastric bypass surgery or restrictive bariatric surgery (such as Lap-Band).

[0188] 12) Has acute or chronic hepatitis, has signs or symptoms of any other liver disease (excluding non-alcoholic fatty liver disease), or has ALT > 5 × ULN at the time of screening;

[0189] 13) Individuals with a personal or family history of medullary thyroid carcinoma (MTC) or multiple endocrine neoplasia type 2 (MEN2), or those with hereditary diseases that predispose them to medullary thyroid carcinoma;

[0190] 14) Individuals with uncontrolled stable thyroid dysfunction (stable control means receiving at least 3 months of stable doses of thyroid hormone replacement or antithyroid drug treatment and having thyroid function within the normal range);

[0191] 15) Those currently suffering from symptomatic cholecystitis or gallstones;

[0192] 16) Any of the following diseases or medical history within the 6 months prior to screening: myocardial infarction, unstable angina, coronary artery bypass grafting, percutaneous coronary intervention (diagnostic angiography is allowed), transient ischemic attack, cerebrovascular accident, or heart failure, etc.

[0193] 17) Poor blood pressure control at the time of screening (systolic blood pressure >160 mmHg or diastolic blood pressure >100 mmHg), or having changed antihypertensive medication within 30 days prior to screening, renal artery stenosis, or evidence of unstable blood pressure, including symptomatic orthostatic hypotension;

[0194] 18) Subjects who have a malignant tumor or a history of malignant tumors, or who are currently being evaluated for potential malignant tumors;

[0195] 19) Those who plan to receive an organ transplant or have already received an organ transplant;

[0196] 20) Known hemoglobinopathies, hemolytic anemia, sickle cell anemia, or at the time of screening, female subjects with hemoglobin <110g / L, male subjects with hemoglobin <120g / L, or any other known condition that interferes with HbA1c detection;

[0197] 21) Individuals who have previously used GLP-1 or FGF21 receptor agonists;

[0198] 22) Use of weight loss drugs within 3 months prior to screening; or chronic (lasting more than 2 weeks) systemic glucocorticoid treatment (excluding topical, intraocular, intranasal, intra-articular or inhaled formulations) within 3 months prior to screening; or systemic glucocorticoid treatment within 1 month prior to screening.

[0199] 23) Subjects who have received the investigational drug or participated in a clinical trial within the three months prior to screening (subjects who withdrew from the study before treatment, i.e., were not randomized or did not receive treatment, may be enrolled in this study);

[0200] 24) Individuals with clinically significant electrocardiogram abnormalities who are deemed unsuitable for enrollment by the researchers;

[0201] 25) Except for type 2 diabetes and fatty liver, any abnormalities in physical examination, vital signs, and laboratory tests of the subjects that are clinically significant and that the investigator judges to pose a significant risk or interference to the subjects will be evaluated for safety, PK, or PD results.

[0202] 26) Women who are breastfeeding or pregnant, or women of childbearing age with a positive pregnancy test;

[0203] 27) Serum calcitonin ≥20 ng / L at screening;

[0204] 28) Fasting triglycerides >500 mg / dL (5.7 mmol / L) at screening time. If the subject is receiving lipid-lowering therapy, the dose of the therapy must remain stable for 30 days prior to screening.

[0205] 29) Creatinine clearance rate <60 mL / min during screening [Calculation formula: CLcr: (140 - age) × weight (kg) / [72 × 0.0113 × Scr (μmol / L)], female × 0.85];

[0206] 30) Positive screening for hepatitis B surface antigen, hepatitis C antibody or core antigen, HIV antibody, and syphilis antibody. [If syphilis antibody is positive, a rapid plasma reagin (RPR) test is required. If RPR is also positive, the candidate must be excluded. If hepatitis C antibody or core antigen is positive, HCV RNA quantification is required. If the subject's HCV RNA result is negative and they have not received anti-HCV treatment within 3 months prior to screening, they can be enrolled.]

[0207] 31) Use of other prohibited concomitant treatments (see section 5.6.2 for details);

[0208] 32) Individuals who test positive for alcohol screening or urine drug screening;

[0209] 33) Subjects who, according to the researchers, have other factors that make them unsuitable to participate in this trial.

[0210] The safety, tolerability, pharmacokinetics, and pharmacodynamics of the FGF21 / GLP-1 fusion protein injection were studied. The results are as follows:

[0211] 1. Pharmacokinetics:

[0212] Serum concentrations of FGF21 / GLP-1, GLP-1 (intact GLP-1 + Fc), and FGF21 (Fc + intact FGF21) will be quantitatively measured separately. Key pharmacokinetic parameters include: peak concentration (C0). max Area under the plasma concentration-time curve (AUC) 0-t AUC 0-∞ Peak time (T) max ), elimination phase half-life (t) 1 / 2 Mean residence time (MRT), clearance rate (CL / F), apparent volume of distribution (Vz / F), elimination rate constant (λz), and AUC 0-∞ Extrapolation percentage (%AUCex), etc.

[0213] In addition, for multiple-dose trials, it is also necessary to obtain the area under the blood drug concentration-time curve (AUC0-τ), percentage of fluctuation (DF), steady-state trough concentration (Css_min), steady-state peak concentration (Css_max), and accumulation ratio (ARCmax or ARAUC) within the dosing interval.

[0214] 2. Pharmacodynamics:

[0215] 1) Standard meal test: Subjects underwent a standard instant noodle meal test on D-1, D30 and D36. Blood samples were collected within 0.5 hours before the standard meal and 0.5 hours, 1 hour, 2 hours, 3 hours and 4 hours after the meal to test blood glucose, insulin, C-peptide and glucagon.

[0216] 2) Fasting blood glucose, fasting blood lipids [triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), TC / HDL-C ratio, apolipoprotein B (ApoB)], alanine aminotransferase (ALT), aspartate aminotransferase (AST): During the screening period, before administration on D8, before administration on D15, before administration on D29, on D38, D43, or before early withdrawal from the visit, the above indicators can be collected during blood biochemistry tests, without the need for additional blood collection.

[0217] 3) HbA1c: during the screening period, D38, or early withdrawal from the visit.

[0218] 4) Adiponectin: Before administration on D-1, D15, D29, or before leaving the visit on D38.

[0219] 5) MRI-PDFF and FibroScan examinations: screening period, D38 (since the MRI-PDFF examination takes a long time, this examination can be completed between D38 and D43) or early withdrawal from the visit.

[0220] 6) Fasting weight: screening period, D-1, D15 before administration, D29 before administration, D38 or early withdrawal from the visit.

[0221] 7) Waist circumference, abdominal circumference, hip circumference, and waist / hip ratio in fasting condition: before administration on D-1, D15, before administration on D29, or before leaving the visit on D38 or earlier.

[0222] 3. Safety:

[0223] The safety and tolerability of FGF21 / GLP-1 fusion protein injection were evaluated in subjects via subcutaneous abdominal injection. Safety evaluation primarily included adverse events, laboratory tests (complete blood count, blood biochemistry, coagulation function, thyroid function, urinalysis), vital signs, physical examination, 12-lead electrocardiogram, bone biomarkers, and serum calcitonin.

[0224] The test results are as follows:

[0225] 1. Pharmacokinetic results:

[0226] Within the dosage range of 5.1 mg to 68.0 mg, the mean plasma concentration-time semi-logarithmic plots of FGF21 / GLP-1, GLP-1, and FGF21 in T2DM subjects at each dosing stage are shown in Figures 1A, 1B, and 1C of Figure 1, respectively.

[0227] The results showed that after 5 weeks of continuous administration to T2DM subjects, FGF21 / GLP-1 reached steady state in the 5.1 mg to 68.0 mg dose groups; GLP-1 and FGF21 reached steady state in the 5.1 mg, 15.3 mg, and 30.6 mg dose groups; in the high-dose group, due to the fewer dosing times of the target dose, GLP-1 had not yet reached steady state in the 45.9 mg and 68.0 mg dose groups, while FGF21 was close to steady state in the 45.9 mg dose group but had not yet reached steady state in the 68.0 mg dose group.

[0228] In patients with type 2 diabetes mellitus (T2DM), 5.1 mg–68.0 mg of FGF21 / GLP-1 was administered subcutaneously via abdominal injection after a meal on day 29. The Tss,max ranges for FGF21 / GLP-1, GLP-1, and FGF21 in each dose group were 12.0–14.0 h, 18.9–48.0 h, and 14.0–31.0 h, respectively; the Css,max ranges were 218.39–7084.52 pmol / L, 821.7–12925.3 pmol / L, and 470.3–9063.7 pmol / L, respectively. AUC 0-168h The ranges were 14959.16–322652.66 h·pmol / L, 83409.3–1218595.0 h·pmol / L, and 42461.1–644544.2 h·pmol / L, respectively.

[0229] In patients with type 2 diabetes mellitus (T2DM), after 5 weeks of continuous administration to reach steady state, FGF21 / GLP-1 showed virtually no accumulation, GLP-1 showed mild accumulation, and FGF21 showed no or slight accumulation.

[0230] In T2DM subjects, 5.1 mg to 68.0 mg of FGF21 / GLP-1 was administered subcutaneously to the abdomen after a meal on day 29. The Vss / F ranges for FGF21 / GLP-1, GLP-1, and FGF21 were 59.22–249.80 L, 42.0–68.6 L, and 39.6–78.0 L, respectively.

[0231] In patients with type 2 diabetes mellitus (T2DM), 5.1 mg–68.0 mg of FGF21 / GLP-1 was administered subcutaneously via the abdomen after a meal on day 29. The t values ​​of FGF21 / GLP-1, GLP-1, and FGF21 were compared between the dose groups. 1 / 2 The ranges are 20.06–25.49 h, 72.8–85.4 h, and 32.5–39.4 h, respectively, and the ranges of CLss / F are 2.05–4.68 L / h, 0.5–0.6 L / h, and 0.8–1.7 L / h, respectively.

[0232] 2. Pharmacodynamic results:

[0233] 1) HbA1c:

[0234] As shown in Figures 2a and 2b, after 5 weeks of continuous administration to T2DM subjects, HbA1c showed a significant decrease from baseline in all FGF21 / GLP-1 dose groups, exhibiting a certain dose-related relationship. The placebo group showed a smaller decrease from baseline in HbA1c. Before placebo adjustment, the average decrease in HbA1c from baseline in all FGF21 / GLP-1 dose groups ranged from -0.48% to -1.10%, with the average decrease in HbA1c from baseline exceeding 1% in the 30.6 mg and 68.0 mg groups. After placebo adjustment, the average decrease in HbA1c from baseline in all FGF21 / GLP-1 dose groups ranged from -0.17% to -0.79%.

[0235] 2) Fasting blood glucose:

[0236] As shown in Figures 2c and 2d, fasting blood glucose levels in all FGF21 / GLP-1 dose groups decreased significantly from baseline. Fasting blood glucose levels in each dose group showed a gradual decreasing trend with prolonged administration, rebounding after discontinuation. The medium- and high-dose groups (30.6 mg–68.0 mg) showed greater reductions in fasting blood glucose from baseline than the low-dose groups (5.1 mg and 15.3 mg). The maximum average reduction in fasting blood glucose from baseline in each FGF21 / GLP-1 dose group ranged from -1.700 mmol / L (15.3 mg group, D36) to -3.139 mmol / L (30.6 mg group, D38). There was no significant change in fasting blood glucose in the placebo group from baseline.

[0237] 3) Instant noodle standard meal test:

[0238] As shown in Figure 3a, in T2DM subjects, blood glucose levels decreased from baseline in all FGF21 / GLP-1 dose groups on days 30 and 36. The decrease in blood glucose from baseline was significantly greater in the medium and high dose groups (30.6 mg to 68.0 mg) than in the low dose group (5.1 mg and 15.3 mg). Blood glucose levels in the placebo group showed no significant change from baseline during the standard meal test.

[0239] 4) Insulin levels:

[0240] Figure 3b shows the linear graph of the mean change in insulin from baseline in subjects with type 2 diabetes mellitus (T2DM). In the medium- and high-dose groups (30.6 mg–68.0 mg), significant increases in insulin from baseline were observed at both D30 and D36. In the placebo group, there was no significant change in insulin from baseline.

[0241] After placebo adjustment, insulin AUC was observed at both D30 and D36 in the medium- and high-dose FGF21 / GLP-1 groups (30.6 mg–68.0 mg). 0-4hSignificantly higher than baseline, with no dose-related effect. D30 insulin AUC 0-4h At a dose of 30.6 mg, the D36 insulin AUC was significantly higher than baseline (P<0.05). 0-4h The insulin AUC was significantly higher than baseline at doses of 30.6 mg and 45.9 mg (P<0.05). 0-4h At the 30.6 mg dose, the largest increase in D30 from baseline was observed, at 591.875 h*pmol / L. In the 30.6 mg group, the D30 insulin AUC... 0-4h The increase from baseline was better than that on D36. In the high-dose groups (45.9 mg, 68.0 mg), the AUC of insulin on D30 was [missing value]. 0-4h The increase from the baseline was slightly less than that of D36.

[0242] 5) Insulin sensitivity and β-cell function:

[0243] Figures 3c and 3d summarize the average percentage change from baseline in insulin sensitivity and β-cell function in T2DM subjects. No significant change in insulin sensitivity from baseline was observed in any of the FGF21 / GLP-1 dose groups and the placebo group on day 30. A significant increase in insulin sensitivity from baseline was observed on day 36. The average percentage increase from baseline was the largest at the 30.6 mg dose, reaching 115.61% (76.87% after placebo correction).

[0244] In all FGF21 / GLP-1 dosage groups, β-cell function showed a significant increase from baseline at days 30 and 36, exhibiting a certain dose-related correlation. The 68.0 mg group showed the largest average percentage increase from baseline at day 30, reaching 152.86% (138.27% after placebo adjustment), with the increase in β-cell function at day 30 being greater than that at day 36. No significant change in β-cell function from baseline was observed in the placebo group.

[0245] 6) Adiponectin:

[0246] As shown in Figure 4, adiponectin levels increased from baseline in all FGF21 / GLP-1 dose groups. During the dosing period, adiponectin levels showed an increasing trend with increasing dose and duration. Before and after placebo correction, the maximum average percentage increase in adiponectin from baseline reached 77.84% (D38 in the 45.9 mg group) and 87.28% (77.84% average percentage change in adiponectin from baseline in the 45.9 mg group on D38 minus the corresponding placebo value), respectively. No significant change in adiponectin from baseline was observed in the placebo group.

[0247] 7) Fasting blood lipids:

[0248] As shown in Figure 5, a significant decrease in triglycerides compared to baseline was observed in all FGF21 / GLP-1 dose groups. Before and after placebo correction, the maximum average percentage decrease in triglycerides compared to baseline reached -42.15% (before administration on D29 in the 68.0 mg group) and -65.85% (the average percentage change in triglycerides compared to baseline before administration on D29 in the 68.0 mg group minus the corresponding placebo value). In the high-dose groups (45.9 mg and 68.0 mg), a decrease in total cholesterol and low-density lipoprotein cholesterol compared to baseline was observed. No significant changes in fasting lipid parameters were observed in the placebo group compared to baseline.

[0249] In the high-dose groups (45.9 mg, 68.0 mg), total cholesterol and low-density lipoprotein cholesterol decreased compared to baseline.

[0250] 8) MRI-PDFF:

[0251] As shown in Figures 6a and 6b, after 5 weeks of continuous administration to T2DM subjects, a significant decrease in MRI-PDFF was observed in all FGF21 / GLP-1 dose groups. The placebo group showed a slight decrease in MRI-PDFF compared to baseline. Before placebo adjustment, the mean percentage decrease in MRI-PDFF from baseline in each FGF21 / GLP-1 dose group ranged from -25.60% to -47.21%. After placebo adjustment, the mean percentage decrease in MRI-PDFF from baseline in each FGF21 / GLP-1 dose group ranged from -10.55% to -32.16%.

[0252] As shown in Figure 6c, in the FGF21 / GLP-1 5.1mg–68.0mg and placebo groups, 4, 6, 6, 7, 5, and 4 subjects had baseline MRI-PDFF ≥8%, respectively. After 5 weeks of continuous administration, 2 (50.0%), 3 (50.0%), 5 (83.3%), 5 (71.4%), 5 (100.0%), and 1 (25.0%) subjects had MRI-PDFF values ​​≥30% lower than baseline, respectively. With increasing dosage, the proportion of subjects with MRI-PDFF values ​​≥30% lower than baseline showed an increasing trend.

[0253] 9) Fasting weight:

[0254] As shown in Figure 6d, after 5 weeks of continuous administration of FGF21 / GLP-1, all dose groups in the T2DM subjects showed a decrease in fasting weight from baseline. The largest decrease in fasting weight from baseline was observed at the 30.6 mg dose on day 38, reaching -2.36 kg (placebo adjusted: -1.40 kg). The placebo group showed a smaller decrease in fasting weight from baseline.

[0255] 10) Liver enzymes:

[0256] As shown in Figure 7, in subjects with type 2 diabetes mellitus (T2DM), a significant decrease in ALT and AST was observed at a dose of 45.9 mg compared to baseline. During the dosing period, the average percentage decrease in ALT and AST compared to baseline was -27.49% to -36.00% and -22.48% to -29.65%, respectively. No significant changes in ALT and AST were observed in the other dose groups and the placebo group compared to baseline.

[0257] After 5 weeks of FGF21 / GLP-1 treatment, liver enzymes, including ALT, AST, and gamma-glutamyl transferase (GGT), showed a decreasing trend. The decreases in ALT and AST were primarily observed in the high-dose groups (45.9 and 68.0 mg). In the 45.9 mg group on day 29, the relative mean changes in ALT and AST reached -36.00% (SD: 25.135) and -29.65% (SD: 22.505), respectively (compared to -18.29% [SD: 29.467] and -14.82% [SD: 15.079] in the placebo group, respectively). A dose-dependent decrease in GGT was observed in most dose groups compared to placebo, with the largest relative mean change reaching -28.30% (SD: 13.776) in the 45.9 mg cohort on day 38 (compared to -11.92% [SD: 25.008] in placebo).

[0258] 3. Safety Results:

[0259] FGF21 / GLP-1 was well-safe and tolerable with multiple doses in subjects with type 2 diabetes mellitus (T2DM) within a dose range of 5.1 mg to 68.0 mg.

[0260] 4. PK and efficacy analysis

[0261] The correlation between the primary efficacy endpoint and drug exposure level was further evaluated. As shown in Figure 8a, the reduction in MRI-PDFF increased with increasing dose. AUC using GLP-1 and FGF21 was also assessed. 0-168h,SS Regression analysis of the decrease in MRI-PDFF from baseline showed that MRI-PDFF may have a further decreasing trend with increasing GLP-1 and FGF21 exposure levels (Figures 8b-c). Similarly, the AUC of GLP-1 and FGF21... 0-168h，ss The increase was also associated with better performance of HbA1c and adiponectin, respectively (Figure 8d-e). The plateau trend of pharmacodynamic parameters in the high-dose group may be attributed to inter-individual variability in the given sample size and exposure level.

[0262] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0263] Although embodiments of this application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting this application. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of this application.

Claims

1. Use of the FGF21 / GLP-1 fusion protein in the preparation of a medicament for the prevention and / or treatment of at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia in individuals in need.

2. The use according to claim 1, characterized in that, The drug is used to reduce at least one of the following indicators: HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol; And / or, the drug is used to improve at least one of the following indicators: Adiponectin, insulin sensitivity, and β-cell function.

3. Use of the FGF21 / GLP-1 fusion protein in the prevention and / or treatment of diseases in individuals in need, said diseases being at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia.

4. The use according to claim 3, characterized in that, The purpose is to reduce at least one of the following indicators: HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol; And / or, the purpose is to improve at least one of the following metrics: Adiponectin, insulin sensitivity, and β-cell function.

5. FGF21 / GLP-1 fusion protein for the prevention and / or treatment of at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia in individuals in need.

6. The use according to claim 5, characterized in that, The FGF21 / GLP-1 fusion protein is used to reduce at least one of the following indicators: HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol; And / or, the FGF21 / GLP-1 fusion protein is used to improve at least one of the following indicators: Adiponectin, insulin sensitivity, and β-cell function.

7. A method for preventing and / or treating a disease in an individual in need, characterized in that, The diseases mentioned include at least one of type 2 diabetes, obesity, fatty liver disease, steatohepatitis, dyslipidemia, and hypertriglyceridemia; The method includes administering a pharmaceutically acceptable dose of the FGF21 / GLP-1 fusion protein to a subject.

8. The method according to claim 7, characterized in that, The prevention and / or treatment are intended to reduce at least one of the following indicators: HbA1c, blood glucose, weight, liver fat content, ALT, AST, gamma-glutamyl transferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol; And / or, the prevention and / or treatment are used to improve at least one of the following indicators: Adiponectin, insulin sensitivity, and β-cell function.

9. The use according to any one of claims 1 to 6 or the method according to any one of claims 7 to 8, characterized in that, The FGF21 / GLP-1 fusion protein has the amino acid sequence shown in SEQ ID NO:1; Optionally, the effective dose of the FGF21 / GLP-1 fusion protein is 5.0 mg to 80.0 mg, preferably 5.0 mg to 68.0 mg; Optionally, the effective dose of the FGF21 / GLP-1 fusion protein is 15.0 mg to 80.0 mg, preferably 15.0 mg to 68.0 mg; Optionally, the effective dose of the FGF21 / GLP-1 fusion protein is 30.0 mg to 80.0 mg, preferably 30.0 mg to 68.0 mg; Optionally, the effective dose of the FGF21 / GLP-1 fusion protein is 45.0 mg to 80.0 mg, preferably 45.0 mg to 68.0 mg; Optionally, the FGF21 / GLP-1 fusion protein is administered in the form of an injection. Optionally, the concentration of the FGF21 / GLP-1 fusion protein in the injection solution is 1–100 mg / mL, 2–80 mg / mL, 5–70 mg / mL, 2–55 mg / mL, 10–65 mg / mL, or 20–60 mg / mL; Optionally, the injection solution further includes at least one of a buffer solution, an osmotic pressure regulator, a surfactant, a preservative, and a metal chelating agent; Optionally, the buffer solution includes at least one of histidine buffer, citrate buffer, phosphate buffer, and Tris buffer.

10. The use or method according to claim 9, characterized in that, The FGF21 / GLP-1 fusion protein is administered via fixed-dose administration or titration administration, preferably titration administration; Optionally, the FGF21 / GLP-1 fusion protein may be administered once a month, once every four weeks, once every three weeks, once every two weeks, once a week, twice a week, three times a week, or once a day.

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