GLP-1-Fc-FGF21 dual-target fusion protein composition, injection solution, and use thereof

Abstract

GLP-1-Fc-FGF21 double-target fusion protein composition and its use. The composition comprises a GLP-1-Fc-FGF21 fusion protein and a buffer solution. In the composition, by adding the buffer solution, the formation of visible particles can be effectively controlled, the stability of the GLP-1-Fc-FGF21 fusion protein can be improved, and the shelf life of the product can be extended. The composition can be used to treat and / or prevent metabolic diseases such as diabetes, dyslipidemia, obesity, and diseases associated with fatty liver.

Description

Technical Field

[0001] [Cross - reference to Related Applications] This application claims the priority and benefit of Chinese Patent Application No. 202310741972.5, filed with the State Intellectual Property Office of China on June 20, 2023, and this Chinese Patent Application No. 202310741972.5 is incorporated herein by reference in its entirety.

[0002] [Field of the Invention] The present invention belongs to the field of biopharmaceutical technology. In particular, the present invention relates to a GLP - 1 - Fc - FGF21 dual - target fusion protein composition, an injection solution, and its use. More specifically, the present invention relates to a composition, an injection solution, use, and an injection device or container.

Background Art

[0003] Glucagon - like peptide - 1 (GLP - 1) is an incretin secreted by L cells in the small intestine. Incretins stimulate pancreatic β - cells to secrete insulin, thereby maintaining insulin balance in the patient's body. GLP - 1 indirectly exerts its effect through insulin and is only effective for type 2 diabetes, which limits the scope of application and efficacy of GLP - 1. Additionally, there are reports suggesting that GLP - 1 may have a potential risk of causing thyroid cancer.

[0004] FGF21 is a member of the fibroblast growth factor (FGF) family. FGF21 has the ability to promote glucose uptake by adipocytes, thereby enhancing insulin sensitivity. Unlike insulin, FGF21 does not induce hypoglycemia or other associated side effects. FGF21 is more effective in protecting beta-islet cells and promoting their regeneration and repair. Furthermore, due to the lack of mitogenic activity in FGF21, it does not pose a potential risk of tumorigenesis. FGF21 shows promise as a therapeutic agent for type 1 diabetes. In addition, FGF21 also exhibits a significant lipid-lowering effect, making it a potential candidate for lipid-lowering drugs.

[0005] The GLP-1-Fc-FGF21 dual-target fusion protein can be used to treat metabolic diseases such as diabetes, dyslipidemia, obesity, and fatty liver disease. However, during storage, this dual-target fusion protein may undergo physical and chemical changes such as denaturation, aggregation, precipitation, or fragmentation, which can affect the stability of the dual-target fusion protein.

[0006] Therefore, there is an urgent need to develop a highly stable GLP-1-Fc-FGF21 dual-target fusion protein formulation. [Overview of the Initiative]

[0007] The present invention aims to solve, to some extent, one of the technical problems present in the prior art. To this end, the present invention provides a GLP-1-Fc-FGF21 dual-target fusion protein composition that exhibits strong stability and can extend the shelf life of the product.

[0008] In one aspect of the present invention, a composition is proposed. According to one embodiment of the present invention, the composition comprises a GLP-1-Fc-FGF21 fusion protein and a buffer solution. In the composition of the present invention, the addition of the buffer solution can effectively control the formation of visible particles and aggregates, enhance the stability of the GLP-1-Fc-FGF21 fusion protein, and thereby extend the shelf life of the product.

[0009] According to one embodiment of the present invention, the GLP-1-Fc-FGF21 fusion protein contains the amino acid sequences shown in SEQ ID NOs: 1 to 12.

[0010] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 1 to 100 mg / mL.

[0011] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 80 mg / mL.

[0012] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 70 mg / mL.

[0013] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 5 to 70 mg / mL.

[0014] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 65 mg / mL.

[0015] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 60 mg / mL.

[0016] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 55 mg / mL.

[0017] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 10 to 65 mg / mL.

[0018] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 20 to 60 mg / mL.

[0019] According to an embodiment of the present invention, the pH value of the composition is 6.2 to 8.0, preferably 6.4 to 8.0.

[0020] According to an embodiment of the present invention, the buffer solution contains at least one of a histidine buffer solution, a citrate buffer solution, a phosphate buffer solution, and a Tris buffer solution.

[0021] According to an embodiment of the present invention, the buffer solution contains a histidine buffer solution or a citrate buffer solution.

[0022] According to an embodiment of the present invention, the concentration of the buffer solution in the composition is 5 to 30 mM.

[0023] According to an embodiment of the present invention, the concentration of the buffer solution in the composition is 5 to 25 mM.

[0024] According to an embodiment of the present invention, the concentration of the buffer solution in the composition is 8 to 25 mM.

[0025] According to an embodiment of the present invention, the concentration of the buffer solution in the composition is 5 to 20 mM.

[0026] According to an embodiment of the present invention, the concentration of the buffer solution in the composition is 8 to 12 mM.

[0027] According to an embodiment of the present invention, the concentration of the buffer solution in the composition is 10 to 20 mM.

[0028] According to an embodiment of the present invention, the composition further contains at least one of an osmotic pressure regulator, a surfactant, a preservative, and a metal chelating agent.

[0029] According to one embodiment of the present invention, the osmotic pressure regulator contains at least one of mannitol, sucrose, sodium chloride, and trehalose.

[0030] According to one embodiment of the present invention, the concentration of the osmotic pressure regulator in the composition is 30 to 100 mg / mL or 1 to 10 mg / mL.

[0031] According to one embodiment of the present invention, the surfactant contains at least one of polysorbate 80, polysorbate 20, and poloxamer 188.

[0032] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 10 mg / mL.

[0033] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL.

[0034] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 6 to 10 mg / mL.

[0035] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 0.7 mg / mL.

[0036] According to one embodiment of the present invention, the preservative is at least one of phenol and m-cresol.

[0037] According to one embodiment of the present invention, the concentration of the preservative in the composition is 0.1 to 10 mg / mL, preferably 0.1 to 5.0 mg / mL, more preferably 1.0 to 5.0 mg / mL.

[0038] According to one embodiment of the present invention, the metal chelating agent is at least one of disodium edetate and calcium disodium edetate.

[0039] According to one embodiment of the present invention, the concentration of the metal chelating agent in the composition is 0.01 to 1 mg / mL, preferably 0.05 to 0.5 mg / mL.

[0040] According to one embodiment of the present invention, the buffer is a histidine buffer, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, or 25 to 85 mg / mL.

[0041] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25 to 80 mg / mL, and the buffer is histidine buffer.

[0042] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25 to 55 mg / mL, and the buffer is histidine buffer.

[0043] According to one embodiment of the present invention, the concentration of the histidine buffer in the composition is 5 to 30 mM.

[0044] According to one embodiment of the present invention, the concentration of the histidine buffer in the composition is 5 to 25 mM.

[0045] According to one embodiment of the present invention, the concentration of the histidine buffer in the composition is 8 to 25 mM.

[0046] According to one embodiment of the present invention, the concentration of the histidine buffer in the composition is 8 to 20 mM.

[0047] According to one embodiment of the present invention, the concentration of the histidine buffer in the composition is 10 to 20 mM.

[0048] According to one embodiment of the present invention, the concentration of the histidine buffer in the composition is 8 to 12 mM.

[0049] According to one embodiment of the present invention, the pH value of the composition is 6.7 to 7.5, preferably 6.9 to 7.5.

[0050] According to one embodiment of the present invention, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30 to 50 mg / mL; or the osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60 to 100 mg / mL, preferably 80 to 100 mg / mL; or the osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40 to 100 mg / mL, preferably 40 to 70 mg / mL, more preferably 50 to 70 mg / mL.

[0051] According to one embodiment of the present invention, the surfactant is polysorbate 80, and the concentration of the surfactant in the composition is 0.1 to 2 mg / mL, preferably 0.1 to 1.5 mg / mL, more preferably 0.4 to 1.3 mg / mL; or the surfactant is polysorbate 20, and the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL, preferably 0.1 to 0.5 mg / mL.

[0052] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 25 mg / mL, and the buffer is a citrate buffer.

[0053] According to one embodiment of the present invention, the concentration of the citrate buffer in the composition is 5 to 30 mM.

[0054] According to one embodiment of the present invention, the concentration of the citrate buffer in the composition is 5 to 25 mM.

[0055] According to one embodiment of the present invention, the concentration of the citrate buffer in the composition is 8 to 25 mM.

[0056] According to one embodiment of the present invention, the concentration of the citrate buffer in the composition is 8 to 20 mM.

[0057] According to one embodiment of the present invention, the concentration of the citrate buffer in the composition is 10 to 20 mM.

[0058] According to one embodiment of the present invention, the concentration of the citrate buffer in the composition is 8 to 12 mM.

[0059] According to one embodiment of the present invention, the pH value of the composition is 6.4 to 8.0.

[0060] According to one embodiment of the present invention, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30 to 50 mg / mL; or the osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60 to 100 mg / mL, preferably 80 to 100 mg / mL; or the osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40 to 100 mg / mL, preferably 40 to 70 mg / mL, more preferably 50 to 70 mg / mL.

[0061] According to one embodiment of the present invention, the osmotic pressure regulator is sodium chloride, and the concentration of the osmotic pressure regulator in the composition is 1 to 10 mg / mL, preferably 3 to 9 mg / mL.

[0062] According to one embodiment of the present invention, the surfactant comprises at least one of polysorbate 80, polysorbate 20, and poloxamer 188, preferably polysorbate 80.

[0063] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 10.0 mg / mL.

[0064] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL.

[0065] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 1.0 mg / mL.

[0066] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 6.0 to 10.0 mg / mL.

[0067] According to one embodiment of the present invention, the concentration of the surfactant in the composition is 0.1 to 0.5 mg / mL.

[0068] According to one embodiment of the present invention, the surfactant is poloxamer 188, and the concentration of the surfactant in the composition is 6 to 10 mg / mL; or the surfactant is polysorbate 80 or polysorbate 20, and the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL, preferably 0.1 to 1.0 mg / mL or 0.1 to 0.5 mg / mL.

[0069] According to one embodiment of the present invention, the dosage form of the composition is an injectable solution.

[0070] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution The concentration of the methyl ester is 25-100 mg / mL; the concentration of histidine buffer in the injection solution is 8-25 mM; the concentration of mannitol in the injection solution is 35-45 mg / mL; the concentration of polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; the concentration of the metal chelating agent in the injection solution is 0.05-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0071] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0072] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, 25-80 mg / mL, 25-75 mg / mL The concentrations are 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 concentration in the composition is 8-25 mM; the mannitol concentration in the composition is 35-45 mg / mL; the polysorbate 80 concentration in the composition is 0.4-1.3 mg / mL, or the polysorbate 20 concentration in the composition is 0.1-0.5 mg / mL; the metal chelating agent concentration in the composition is 0.05-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0073] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0074] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in SEQ ID NO: 1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25-5 The concentration of the histidine buffer in the injection solution is 0 mg / mL; the concentration of the mannitol in the injection solution is 10-20 mM; the concentration of the mannitol in the injection solution is 35-45 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; the concentration of the metal chelating agent in the injection solution is 0.05-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0075] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0076] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the GLP-1-Fc-FGF21 fusion protein in the injection solution The concentration of the protein is 25-100 mg / mL; the concentration of histidine buffer in the injection solution is 8-25 mM; the concentration of sucrose in the injection solution is 85-95 mg / mL, or the concentration of trehalose in the injection solution is 55-65 mg / mL; the concentration of polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0077] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0078] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, or 25-80 mg / mL. The concentrations are 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 concentration in the composition is 8-20 mM; the sucrose concentration in the composition is 85-95 mg / mL; the trehalose concentration in the injection is 55-65 mg / mL; the polysorbate 80 concentration in the composition is 0.4-1.3 mg / mL, or the polysorbate 20 concentration in the composition is 0.1-0.5 mg / mL; and the pH of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0079] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0080] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL; the concentration of the histidine buffer in the injection solution is 10-20 mM; the concentration of the sucrose in the injection solution is 85-95 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0081] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0082] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, trehalose, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL; the concentration of the histidine buffer in the injection solution is 10-20 mM; the concentration of the trehalose in the injection solution is 55-65 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0083] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0084] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, or 25 to 80 mg / mL The concentration of the citrate buffer in the injection solution is 8-25 mM, preferably 10-20 mM; the concentration of sucrose in the injection solution is 85-95 mg / mL; the concentration of trehalose in the injection solution is 55-65 mg / mL; the concentration of polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0085] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0086] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, The concentration of the methyl phosphate is 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 citrate buffer in the injection is 8-25 mM, preferably 10-20 mM; the concentration of mannitol in the injection is 35-45 mg / mL; the concentration of polysorbate 80 in the injection is 0.4-1.5 mg / mL or 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the injection is 0.1-0.5 mg / mL; the concentration of the metal chelating agent in the injection is 0.05-0.5 mg / mL; and the pH of the injection is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0087] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate. In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 5 to 20 mM; the concentration of the mannitol in the injection solution is 40 to 50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 1.5 mg / mL; the concentration of the metal chelating agent in the injection solution is 0.05 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0088] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0089] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of the mannitol in the injection solution is 40 to 50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 0.5 mg / mL; the concentration of the metal chelating agent in the injection solution is 0.05 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0090] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0091] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of the mannitol in the injection solution is 40 to 50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 0.5 mg / mL; the concentration of the metal chelating agent in the injection solution is 0.05 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0092] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0093] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80, and optionally disodium edetate or calcium disodium edetate, and sucrose or trehalose; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 5 to 20 mM; the concentration of the sucrose in the injection solution is 80 to 95 mg / mL; the concentration of the trehalose in the injection solution is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 1.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0094] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0095] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80, and sucrose or trehalose, and optionally disodium edetate or calcium 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 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of the sucrose in the injection solution is 80 to 95 mg / mL; the concentration of the trehalose in the injection solution is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0096] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0097] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0098] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0099] In one optional embodiment of the present invention, the concentration of phenol in the composition or injection solution is 0 to 7 mg / mL, preferably 3 to 7 mg / mL.

[0100] In one optional embodiment of the present invention, the concentration of m-cresol in the composition or injection solution is 0 to 4 mg / mL, preferably 2 to 4 mg / mL.

[0101] In another aspect of the present invention, the present invention provides the use of the aforementioned compositions or injectable solutions in the preparation of pharmaceuticals for treating and / or preventing metabolic diseases.

[0102] According to one embodiment of the present invention, metabolic diseases include diabetes, dyslipidemia, obesity, and diseases associated with fatty liver.

[0103] According to one embodiment of the present invention, diseases associated with fatty liver include non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis.

[0104] In another aspect of the present invention, the present invention provides an injection device or container. According to one embodiment of the present invention, the injection device or container contains the aforementioned composition or the aforementioned injection solution.

[0105] According to one embodiment of the present invention, the injection device or container includes at least one selected from a vial and a syringe.

[0106] According to one embodiment of the present invention, the syringe includes at least one selected from an auto-injector and a pre-filled syringe.

[0107] Additional aspects and advantages of the present invention are partially expressed in the following description, will be partially evident from the following description, or can be learned through the practice of the present invention.

[0108] The above and / or additional aspects and advantages of the present invention will become apparent and readily apparent from the description of the embodiments in conjunction with the accompanying drawings. [Brief explanation of the drawing]

[0109] [Figure 1] This figure shows the high-temperature SEC (size exclusion chromatography) aggregate growth trend chart for various formulations in step 7 of Example 1 of the present invention. [Figure 2]This figure shows the growth trend chart of high-temperature RP-HPLC (reverse-phase high-performance liquid chromatography) purity (total related substances) for various formulations in step 7 of Example 1 of the present invention. [Modes for carrying out the invention]

[0110] Embodiments of the present invention are described in detail below. The embodiments described below are illustrative and intended solely to illustrate the present invention and should not be construed as limiting the invention.

[0111] It should be noted that the terms “first” and “second” are used solely for descriptive purposes and should not be interpreted as indicating or implying relative importance, or implicitly specifying the quantity of the technical characteristics described. Therefore, characteristics designated as “first” or “second” may explicitly or implicitly include one or more such characteristics. Furthermore, in the description of this invention, unless otherwise specified, the term “multiple” means two or more.

[0112] The endpoints and arbitrary values ​​of the domains disclosed herein are not limited to exact domains or values, and these domains or values ​​should be understood to include values ​​close to them. With respect to domains, one or more new domains can be obtained by combining the endpoint values ​​of each domain, the endpoint values ​​of each domain and the values ​​of individual points, and the values ​​of individual points, and these new domains should be considered to be specifically disclosed herein.

[0113] In this document, the terms “comprise” or “include” are unrestricted and mean that they include the content specified by the present invention but do not exclude other content.

[0114] In this document, the terms "optional," "optional," or "optional" generally mean that the events or conditions described later may occur but do not necessarily occur, and the descriptions include the circumstances under which the events or conditions occur, and the circumstances under which they do not occur.

[0115] In this document, the term “treatment” means achieving a desired pharmacological and / or physiological effect. The effect may be prophylactic in the sense of completely or partially preventing a disease or its symptoms, and / or therapeutic in the sense of partially or completely curing a disease and / or adverse effects caused by the disease. As used herein, “treatment” encompasses diseases in mammals, particularly humans, and includes (a) preventing the onset of a disease or condition in an individual susceptible to the disease but not yet diagnosed as having the disease; (b) suppressing the disease, such as halting its progression; or (c) alleviating the disease, such as reducing symptoms associated with the disease. As used herein, “treatment” encompasses any administration of a composition, injection, or drug to an individual for the purpose of treating, curing, alleviating, improving, reducing, or suppressing a disease in an individual, including but not limited to administering a drug containing a composition or injection solution described herein to an individual in need.

[0116] The present invention proposes compositions, injectable solutions, uses thereof, and injection devices or containers, as described in detail below.

[0117] composition In one aspect of the present invention, a composition is proposed. According to one embodiment of the present invention, the composition comprises a GLP-1-Fc-FGF21 fusion protein and a buffer solution. In the composition of the present invention, the addition of the buffer solution can effectively control the formation of visible particles and aggregates, enhance the stability of the GLP-1-Fc-FGF21 fusion protein, and thereby extend the shelf life of the product.

[0118] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequences shown in SEQ ID NOs: 1 to 12.

[0119] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(Sequence ID 1);

[0120] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(Sequence ID 2);

[0121] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPAVFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(Sequence ID 3);

[0122] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYES(Sequence ID 4);

[0123] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTECHLEIREDGTVGCAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(Sequence ID 5);

[0124] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTECHLEIREDGTVGCAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(Sequence ID 6);

[0125] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYES(Sequence ID 7);

[0126] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(Sequence ID 8);

[0127] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(Sequence ID 9);

[0128] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQQYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPAQFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(Sequence ID 10);

[0129] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQQYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPAQFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(Sequence ID 11);

[0130] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKP REEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYES(Sequence ID 12)

[0131] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequences shown in SEQ ID NOs: 1 to 12.

[0132] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1.

[0133] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 2.

[0134] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 3.

[0135] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 4.

[0136] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 5.

[0137] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 6.

[0138] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 7.

[0139] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 8.

[0140] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 9.

[0141] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 10.

[0142] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 11.

[0143] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 12.

[0144] In some optional embodiments of the present invention, the concentration of GLP-1-Fc-FGF21 fusion protein in the composition is 1 to 100 mg / mL, for example, 1 mg / mL, 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, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 8 5 mg / mL, 90 mg / mL, 100 mg / mL, etc., or any threshold between two of these values ​​as the endpoint, e.g., 2-99 mg / mL, 2-95 mg / mL, 2-90 mg / mL, 2-85 mg / mL, 2-80 mg / mL, 2-75 mg / mL, 2-70 mg / mL, 2-65 mg / mL, 2-60 mg / mL, 2-55 mg / mL, 5-99 mg / mL, 5-95 mg / mL, 5-90 mg / mL, 5-85 mg / mL, 5-80 mg / mL, 5-75 mg / mL, 5-70 mg / mL, 5-65 mg / mL, 5-60 mg / mL, 5-55 mg / mL g / mL, 10~99mg / mL, 10~95mg / mL, 10~90mg / mL, 10~85mg / mL, 10~80mg / mL, 10~75mg / mL, 10~70mg / mL, 10~65mg / mL, 10~60mg / mL, 10~55mg / mL, 25~99mg / mL , 25~95mg / mL, 25~90mg / mL, 25~85mg / mL, 25~80mg / mL, 25~75mg / mL, 25~70mg / mL, 25~65mg / mL, 25~60mg / mL, 25~55mg / mL, 30~99mg / mL, 30~95mg / mL, 30~ 90mg / mL, 30~85mg / mL, 30~80mg / mL, 30~75mg / mL, 30~70mg / mL, 30~65mg / mL, 30~60mg / mL, 30~55mg / mL, 35~99mg / mL, 35~95mg / mL, 35~90mg / mL, 35~85mg / mL, 35~80mg / mL, 35~75mg / mL, 35~70mg / mL, 35~65mg / mL, 35~60mg / mL, 35~55mg / mL, 40~99mg / mL, 40~95mg / mL, 40~90mg / mL, 40~85mg / mL, 40~80mg / mL,These ranges from 40-75 mg / mL, 40-70 mg / mL, 40-65 mg / mL, 40-60 mg / mL, 40-55 mg / mL, etc.

[0145] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 55 mg / mL, for example, 2 to 25 mg / mL, 3 to 25 mg / mL, 10 to 25 mg / mL, 10 to 20 mg / mL, 15 to 25 mg / mL, 15 to 20 mg / mL, 25 to 100 mg / mL, 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, 25 to 70 mg / mL, 25 to 65 mg / mL, 25 to 60 mg / mL, 25 to 55 mg / mL, 25 to 50 mg / mL, 25 to 40 mg / mL, 40 to 55 mg / mL, 40 to 50 mg / mL, 45 to 55 mg / mL, or 45 to 50 mg / mL.

[0146] In some optional embodiments of the present invention, the pH value of the composition is 6.4 to 8.0, for example, 6.4, 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, or 8.0, or any threshold between two of these values ​​as an endpoint. This pH value can further enhance the stability of the fusion protein, for example, the pH value of the composition is 6.2 to 8.0 or 6.9 to 7.5.

[0147] In some optional embodiments of the present invention, the buffer solution comprises at least one of histidine buffer, citrate buffer, phosphate buffer, and Tris buffer.

[0148] In this document, unless otherwise specified, “citrate buffer” or “citrate buffer solution” may be selected from citric acid, sodium citrate, or a mixture of citric acid and sodium citrate, all of which fall within the scope of protection of this application.

[0149] In some optional embodiments of the present invention, the phosphate buffer is selected from a combination of disodium hydrogen phosphate and sodium dihydrogen phosphate, or a combination of dipotassium hydrogen phosphate and potassium dihydrogen phosphate.

[0150] In some optional embodiments of the present invention, the histidine buffer comprises histidine and / or histidine hydrochloride.

[0151] In some optional embodiments of the present invention, the histidine buffer comprises histidine and histidine hydrochloride.

[0152] In some optional embodiments of the present invention, the histidine buffer comprises histidine or histidine hydrochloride.

[0153] In some optional embodiments of the present invention, the citrate buffer comprises citric acid and / or sodium citrate.

[0154] In some optional embodiments of the present invention, the citrate buffer comprises citrate and sodium citrate.

[0155] In some optional embodiments of the present invention, the citrate buffer comprises citrate or sodium citrate.

[0156] In some optional embodiments of the present invention, the Tris buffer comprises Tris and / or Tris-HCl.

[0157] In some optional embodiments of the present invention, the Tris buffer comprises Tris and Tris-HCl.

[0158] In some optional embodiments of the present invention, the Tris buffer comprises Tris or Tris-HCl.

[0159] In some optional embodiments of the present invention, the buffer comprises a histidine buffer or a citrate buffer.

[0160] In some optional embodiments of the present invention, the concentration of the buffer in the composition is 5 to 30 mM, for example 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 11.29 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, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, or 30 mM, or any threshold between two of these values ​​as an endpoint (for example 5 to 25 mM, 8 to 25 mM, 8 to 12 mM, 10 to 20 mM).

[0161] In some optional embodiments of the present invention, the composition further comprises at least one of an osmotic pressure regulator, a surfactant, a preservative, and a metal chelating agent. This at least one may further enhance the stability of the fusion protein.

[0162] In some optional embodiments of the present invention, the osmotic pressure regulator comprises at least one of mannitol, sucrose, sodium chloride, and trehalose.

[0163] In some optional embodiments of the present invention, the concentration of the osmotic regulator in the composition is 30 to 100 mg / mL, for example, 30 mg / mL, 35 mg / mL, 40 mg / mL, 41 mg / mL, 41.4 mg / mL, 41.5 mg / mL, 42 mg / mL, 42.4 mg / mL, 42.5 mg / mL, 43 mg / mL, 43.4 mg / mL, 43.5 mg / mL, 44 mg / mL, 44.4 mg / mL, 44.5 mg / mL, 45 mg / mL, 45.4 mg / mL, 45.5 mg / mL, 46 mg / mL, 46 0.4 mg / mL, 46.5 mg / mL, 47 mg / mL, 47.4 mg / mL, 47.5 mg / mL, 48 mg / mL, 48.4 mg / mL, 48.5 mg / mL, 49 mg / mL, 49.4 mg / mL, 49.5 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, or 100 mg / mL, etc., or any threshold between two of these values ​​as the endpoint.

[0164] In some optional embodiments of the present invention, the concentration of the osmotic regulator in the composition is 1 to 10 mg / mL, for example, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, or 10 mg / mL, or any threshold between two of these values ​​as an endpoint.

[0165] In some optional embodiments of the present invention, the surfactant comprises at least one of polysorbate 80, polysorbate 20, and poloxamer 188.

[0166] In some optional embodiments of the present invention, the concentration of the surfactant in the composition is 0.1 to 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, 1.4 mg / mL, 1.5 mg / mL, 1.6 mg / mL, 1.7 mg / mL, 1.8 mg / mL, 1.9 mg / mL. g / mL, 2.0 mg / mL, 2.5 mg / mL, 3 mg / mL, 3.5 mg / mL, 4 mg / mL, 4.5 mg / mL, 5 mg / mL, 5.5 mg / mL, 6 mg / mL, 6.5 mg / mL, 7 mg / mL, 7.5 mg / mL, 8 mg / mL, 8.5 mg / mL, 9 mg / mL, 9.5 mg / mL, or 10 mg / mL, etc., or any threshold between two of these values ​​as the endpoint (e.g., 1 to 1.5 mg / mL, 6 to 10 mg / mL, 0.1 to 0.7 mg / mL).

[0167] For example, the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL or 0.1 to 1 mg / mL.

[0168] In some optional embodiments of the present invention, the preservative comprises at least one of phenol and m-cresol.

[0169] According to embodiments of the present invention, the concentration of the preservative in the composition is 0.1 to 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, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, 7.5 mg / mL, 8.0 mg / mL, 8.5 mg / mL, 9.0 mg / mL, 9.5 mg / mL, or 10.0 mg / mL, for example, 1.0 to 5.0 mg / mL.

[0170] In some optional embodiments of the present invention, the preservative is phenol.

[0171] According to embodiments of the present invention, the concentration of phenol in the composition is 1.0 to 5.0 mg / mL, for example 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, 7.5 mg / mL, 8.0 mg / mL, 8.5 mg / mL, 9.0 mg / mL, 9.5 mg / mL, or 10.0 mg / mL, for example 2 to 7 mg / mL or 3.0 to 4.0 mg / mL.

[0172] In some optional embodiments of the present invention, the preservative is m-cresol.

[0173] According to embodiments of the present invention, the concentration of m-cresol in the composition is 1.0 to 5.0 mg / mL, for example 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, 7.5 mg / mL, 8.0 mg / mL, 8.5 mg / mL, 9.0 mg / mL, 9.5 mg / mL, or 10.0 mg / mL, for example 3.0 to 4.0 mg / mL.

[0174] In some optional embodiments of the present invention, the preservative comprises both phenol and m-cresol.

[0175] According to embodiments of the present invention, the concentration of phenol in the composition is 1.0 to 5.0 mg / mL (for example, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, 7.5 mg / mL, 8.0 mg / mL) The concentration of m-cresol in the composition is 0.1 to 1.0 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, or 1.0 mg / mL).

[0176] In some optional embodiments of the present invention, the metal chelating agent comprises at least one of disodium edetate (EDTA-2Na) and calcium disodium edetate (EDTA-CaNa2).

[0177] In some optional embodiments of the present invention, the metal chelating agent is disodium edetate (EDTA-2Na).

[0178] In some optional embodiments of the present invention, the metal chelating agent is calcium disodium edetate (EDTA-CaNa2).

[0179] In some optional embodiments of the present invention, the concentration of the metal chelating agent in the composition is 0.01 to 1 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, 0.1 mg / mL, 0.11 mg / mL, 0.115 mg / mL, 0.12 mg / mL, 0.13 mg / mL, 0.14 mg / mL, 0.15 mg / mL, 0.16 mg / mL, 0.17 mg / mL, 0.18 mg / mL, 0.19 mg / mL. g / mL, 0.2 mg / mL, 0.25 mg / mL, 0.3 mg / mL, 0.35 mg / mL, 0.4 mg / mL, 0.45 mg / mL, 0.5 mg / mL, 0.55 mg / mL, 0.6 mg / mL, 0.65 mg / mL, 0.7 mg / mL, 0.75 mg / mL, 0.8 mg / mL, 0.85 mg / mL, 0.9 mg / mL, 0.95 mg / mL, or 1.0 mg / mL, etc., or any threshold between two of these values ​​as the endpoint (e.g., 0.05~0.5 mg / mL, 0.08~0.12 mg / mL, 0.1~0.12 mg / mL).

[0180] Exemplary, the concentration of the metal chelating agent in the composition is 0.08 to 0.12 mg / mL. In embodiments of the present invention, the stability of the fusion protein formulation remains good regardless of whether EDTA sodium salt is added, whether it is a citrate-based or histidine buffer system. Therefore, both encapsulation and exclusion of EDTA sodium salt fall within the scope of protection of the present invention.

[0181] In some optional embodiments of the present invention, the buffer is histidine buffer, and the concentration of GLP-1-Fc-FGF21 fusion protein in the composition is 25 to 100 mg / mL, for example 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, or 100 mg / mL, or any range between two of these values ​​as an endpoint. The values ​​are (for example, 25-75 mg / mL, 25-70 mg / mL, 25-65 mg / mL, 25-60 mg / mL, 25-55 mg / mL, 25-50 mg / mL, 25-40 mg / mL, 40-80 mg / mL, 40-75 mg / mL, 40-70 mg / mL, 40-65 mg / mL, 40-60 mg / mL, 40-55 mg / mL, 40-50 mg / mL, 45-80 mg / mL, 45-75 mg / mL, 45-70 mg / mL, 45-65 mg / mL, 45-60 mg / mL, 45-55 mg / mL, or 45-50 mg / mL).

[0182] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25-55 mg / mL, and the buffer is histidine buffer. The inventors unexpectedly discovered that when the final concentration of the fusion protein is 25-55 mg / mL, the addition of histidine buffer can effectively control the formation of visible particles in the composition. This discovery not only improves the physical stability of the fusion protein and reduces potential safety risks, but also effectively controls the rate of chemical change, maintaining stability under high temperature, accelerated, long-term, freeze-thaw, shaking, and simulated transport conditions. The resulting composition has an extended storage life.

[0183] In some optional embodiments of the present invention, the concentration of the histidine buffer in the composition is 5 to 30 mM, for example 5 mM, 6 mM, 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, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, or 30 mM, or any threshold between two of these values ​​as an endpoint (for example 5 to 25 mM, 8 to 25 mM, 8 to 20 mM, 10 to 20 mM, or 8 to 12 mM).

[0184] In some optional embodiments of the present invention, the concentration of histidine buffer in the composition is 10 to 20 mM.

[0185] In some optional embodiments of the present invention, the pH value of the composition is 6.7 to 7.5, preferably 6.9 to 7.5.

[0186] In some optional embodiments of the present invention, the osmoregulator is mannitol, and the concentration of mannitol in the composition is 30 to 50 mg / mL; or the osmoregulator is sucrose, and the concentration of sucrose in the composition is 60 to 100 mg / mL, preferably 80 to 100 mg / mL; or the osmoregulator is trehalose, and the concentration of trehalose in the composition is 40 to 100 mg / mL, preferably 40 to 70 mg / mL, 50 to 70 mg / mL, or 55 to 65 mg / mL; or the osmoregulator is sodium chloride, and the concentration of sodium chloride in the composition is 1 to 10 mg / mL, preferably 3 to 9 mg / mL.

[0187] In some optional embodiments of the present invention, the surfactant is polysorbate 80, and the concentration of polysorbate 80 in the composition is 0.1 to 2 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, 1.4 mg / mL, 1.5 mg / mL, 1.6 mg / mL, 1.7 mg / mL, 1.8 mg / mL, 1.9 mg / mL, or 2.0 mg / mL, or any threshold between two of these values ​​as an endpoint (e.g., 0.1 to 1.5 mg / mL, 0.2 to 2 mg / mL, or 0.4 to 1.3 mg / mL); or In some optional embodiments of the present invention, the surfactant is polysorbate 20, and the concentration of polysorbate 20 in the composition is 0.1 to 1.5 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, 1.4 mg / mL, or 1.5 mg / mL, or any threshold between two of these values ​​as an endpoint (e.g., 0.1 to 0.5 mg / mL).

[0188] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 25 mg / mL, and the buffer is a citrate buffer. The inventors unexpectedly discovered that when the final concentration of the fusion protein is 2 to 25 mg / mL, the addition of a citrate buffer can effectively control the formation of visible particles in the composition. This discovery improves the physical and chemical stability of the fusion protein, reduces potential safety risks, and extends the shelf life of the composition.

[0189] In some optional embodiments of the present invention, the concentration of the citrate buffer in the composition is 5 to 30 mM, for example 5 mM, 6 mM, 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, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, or 30 mM, or any threshold between two of these values ​​as an endpoint (for example 5 to 25 mM, 8 to 25 mM, 8 to 20 mM, 10 to 20 mM, 8 to 12 mM, 5 to 15 mM, or 10 to 15 mM).

[0190] In some optional embodiments of the present invention, the pH value of the composition is 6.4 to 8.0.

[0191] In some optional embodiments of the present invention, the osmotic regulator is mannitol, and the concentration of mannitol in the composition is 30 to 50 mg / mL; or the osmotic regulator is sucrose, and the concentration of sucrose in the composition is 60 to 100 mg / mL, preferably 80 to 100 mg / mL; or the osmotic regulator is trehalose, and the concentration of trehalose in the composition is 40 to 100 mg / mL, preferably 40 to 70 mg / mL, more preferably 50 to 70 mg / mL.

[0192] According to one embodiment of the present invention, the osmotic pressure regulator is sodium chloride, and the concentration of the osmotic pressure regulator in the composition is 1 to 10 mg / mL, preferably 3 to 9 mg / mL.

[0193] In some optional embodiments of the present invention, the surfactant comprises at least one of polysorbate 80, polysorbate 20, and poloxamer 188, preferably polysorbate 80.

[0194] In some optional embodiments of the present invention, the concentration of the surfactant in the composition is 0.1 to 10.0 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, or 10.0 mg / mL, or any threshold between two of these values ​​as an endpoint (for example, 0.1 to 1.5 mg / mL, 0.1 to 1.0 mg / mL, 6.0 to 10.0 mg / mL, 0.1 to 0.5 mg / mL, or 0.1 to 0.3 mg / mL).

[0195] In some optional embodiments of the present invention, the surfactant is poloxamer 188, and the concentration of poloxamer 188 in the composition is 6 to 10 mg / mL; or the surfactant is polysorbate 80 or polysorbate 20, and the concentration of polysorbate 80 or polysorbate 20 in the composition is 0.1 to 1.5 mg / mL, preferably 0.1 to 1 mg / mL or 0.1 to 0.5 mg / mL.

[0196] In some optional embodiments of the present invention, the dosage form of the composition is an injectable formulation.

[0197] injection solution In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 2 The concentration of the histidine buffer in the composition is 5-100 mg / mL; the concentration of the mannitol in the composition is 8-25 mM; the concentration of the mannitol in the composition is 35-45 mg / mL; the concentration of the polysorbate 80 in the composition is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the composition is 0.1-0.5 mg / mL; the concentration of the metal chelating agent in the composition is 0.05-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the formation of visible particles and aggregates, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0198] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-95 mg / mL.

[0199] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-90 mg / mL.

[0200] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-85 mg / mL.

[0201] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-80 mg / mL.

[0202] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-75 mg / mL.

[0203] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-70 mg / mL.

[0204] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-65 mg / mL.

[0205] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-60 mg / mL.

[0206] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-55 mg / mL.

[0207] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL.

[0208] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 12.

[0209] According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in SEQ ID NO: 1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL; the concentration of the histidine buffer in the composition is 10-20 mM; the concentration of the mannitol in the composition is 35-45 mg / mL; the concentration of the polysorbate 80 in the composition is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the composition is 0.1-0.5 mg / mL; the concentration of the metal chelating agent in the composition is 0.05-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5.

[0210] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0211] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0212] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0213] In one optional embodiment of this application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 3 to 7 mg / mL, 2 to 4 mg / mL).

[0214] In another optional embodiment of this application, the concentration of m-cresol in the composition or injection is 0 to 4 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 2 to 4 mg / mL).

[0215] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose, optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the GLP-1-Fc-FGF21 fusion protein in the injection solution The concentration of the protein is 25-100 mg / mL; the concentration of histidine buffer in the composition is 8-20 mM; the concentration of sucrose in the composition is 85-95 mg / mL; the concentration of trehalose in the injection solution is 55-65 mg / mL; the concentration of polysorbate 80 in the composition is 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the composition is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the formation of visible particles and aggregates, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0216] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-95 mg / mL.

[0217] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-90 mg / mL.

[0218] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-85 mg / mL.

[0219] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-80 mg / mL.

[0220] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-75 mg / mL.

[0221] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-70 mg / mL.

[0222] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-65 mg / mL.

[0223] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-60 mg / mL.

[0224] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-55 mg / mL.

[0225] According to one embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL.

[0226] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 12.

[0227] According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL; the concentration of the histidine buffer in the injection solution is 10-20 mM; the concentration of the sucrose in the injection solution is 85-95 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5.

[0228] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0229] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0230] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0231] In one optional embodiment of this application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 3 to 7 mg / mL, 2 to 4 mg / mL).

[0232] In another optional embodiment of this application, the concentration of m-cresol in the composition or injection is 0 to 4 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, or any range between two of these values ​​as an endpoint (e.g., 2 to 4 mg / mL).

[0233] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, or 25 to 80 mg / mL The concentration of the citrate buffer in the injection solution is 8-25 mM, preferably 10-20 mM; the concentration of sucrose in the injection solution is 85-95 mg / mL; the concentration of trehalose in the injection solution is 55-65 mg / mL; the concentration of polysorbate 80 in the injection solution is 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1-0.5 mg / mL; and the pH of the injection solution is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0234] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0235] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0236] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0237] In one optional embodiment of this application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 3 to 7 mg / mL, 2 to 4 mg / mL).

[0238] In another optional embodiment of this application, the concentration of m-cresol in the composition or injection is 0 to 4 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 2 to 4 mg / mL).

[0239] In another aspect of the present invention, the present invention provides an injectable solution. According to one embodiment of the present invention, the injectable solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injectable solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, The concentration of the methyl phosphate is 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 citrate buffer in the injection is 8-25 mM, preferably 10-20 mM; the concentration of mannitol in the injection is 35-45 mg / mL; the concentration of polysorbate 80 in the injection is 0.4-1.5 mg / mL or 0.4-1.3 mg / mL, or the concentration of polysorbate 20 in the injection is 0.1-0.5 mg / mL; the concentration of the metal chelating agent in the injection is 0.05-0.5 mg / mL; and the pH of the injection is 6.9-7.5. The injection solution of the present invention can effectively control the generation of visible particles, improve the physical and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0240] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0241] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0242] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0243] In one optional embodiment of this application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 3 to 7 mg / mL, 2 to 4 mg / mL).

[0244] In another optional embodiment of this application, the concentration of m-cresol in the composition or injection is 0 to 4 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 2 to 4 mg / mL).

[0245] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium disodium edetate; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the composition is 5 to 20 mM; the concentration of the mannitol in the composition is 40 to 50 mg / mL; the concentration of the polysorbate 80 in the composition is 0.1 to 1.5 mg / mL; the concentration of the metal chelating agent in the composition is 0.05 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the formation of visible particles and aggregates, improve the physical and chemical stability of fusion proteins, thereby reducing potential safety risks and extending the shelf life.

[0246] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 12.

[0247] In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection solution is 8 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection solution is 10 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection solution is 10 to 15 mM.

[0248] According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium 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 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 5 to 25 mM, 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of the mannitol in the injection solution is 40 to 50 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 0.5 mg / mL; the concentration of the metal chelating agent in the injection solution is 0.05 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0.

[0249] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0250] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0251] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0252] In one optional embodiment of this application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 3 to 7 mg / mL, 2 to 4 mg / mL).

[0253] In another optional embodiment of this application, the concentration of m-cresol in the composition or injection is 0 to 4 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 2 to 4 mg / mL).

[0254] In another aspect of the present invention, the present invention provides an injection solution. According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80, optionally disodium edetate or calcium disodium edetate, and sucrose or trehalose; the GLP-1-Fc-FGF21 fusion protein has the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the composition is 5 to 20 mM; the concentration of the sucrose in the composition is 85 to 95 mg / mL; the concentration of the trehalose in the composition is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the composition is 0.1 to 1.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0. The injectable solution of the present invention can effectively control the formation of visible particles and aggregates, improve the physical and chemical stability of fusion proteins, thereby reducing potential safety risks and extending the shelf life.

[0255] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein includes the amino acid sequence shown in SEQ ID NO: 12.

[0256] In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection solution is 8 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection solution is 10 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection solution is 10 to 15 mM.

[0257] According to one embodiment of the present invention, the injection solution comprises a GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80, sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection solution is 5 to 25 mM, 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of the sucrose in the injection solution is 80 to 95 mg / mL; the concentration of the trehalose in the injection solution is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection solution is 0.1 to 0.5 mg / mL; and the pH of the injection solution is 6.4 to 8.0.

[0258] According to one embodiment of the present invention, the injection solution does not contain disodium edetate or calcium disodium edetate.

[0259] In one optional embodiment of the present invention, the injection solution described in the above-described embodiment may further contain a preservative.

[0260] In one optional embodiment of the present invention, the preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0261] In one optional embodiment of this application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, 4.5 mg / mL, 5.0 mg / mL, 5.5 mg / mL, 6.0 mg / mL, 6.5 mg / mL, 7.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 3 to 7 mg / mL, 2 to 4 mg / mL).

[0262] In another optional embodiment of this application, the concentration of m-cresol in the composition or injection is 0 to 4 mg / mL, for example 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL, 4.0 mg / mL, etc., or any range between two of these values ​​as an endpoint (e.g., 2 to 4 mg / mL).

[0263] use In another aspect of the present invention, the present invention provides the use of the aforementioned compositions or injectable solutions in the preparation of pharmaceuticals for treating and / or preventing metabolic diseases.

[0264] In some optional embodiments of the present invention, metabolic diseases include diseases associated with diabetes, obesity, dyslipidemia, and fatty liver.

[0265] In some optional embodiments of the present invention, diseases associated with fatty liver include non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), hepatic fibrosis, and cirrhosis.

[0266] Methods for treating and / or preventing metabolic diseases In another aspect of the present invention, a method for treating and / or preventing metabolic diseases is proposed. According to one embodiment of the present invention, the method comprises the step of administering a pharmaceutically acceptable amount of the aforementioned composition or injection solution to a subject.

[0267] The effective dose of the composition or injection solution described in the present invention may vary depending on the mode of administration, the severity of the disease being treated, and other factors. The selection of a preferred effective dose can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to, pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, and half-life; the severity of the disease being treated, the patient's weight, the patient's immune status, and the route of administration. For example, depending on the urgency of the treatment situation, administration may be carried out at doses once daily, once every other day, once a week, or once every two weeks.

[0268] In some optional embodiments of the present invention, the route of administration of the method includes subcutaneous injection or intravenous injection, preferably subcutaneous injection.

[0269] Injection device or container In another aspect of the present invention, the present invention provides an injection device or container. According to one embodiment of the present invention, the injection device or container contains the aforementioned composition or the aforementioned injection solution.

[0270] In some optional embodiments of the present invention, the injection device or container includes at least one selected from vials, bottles, cartridges, and syringes.

[0271] In some optional embodiments of the present invention, the syringe includes at least one selected from an autoinjector and a prefilled syringe. [Examples]

[0272] The following examples, along with specific embodiments, will illustrate the solutions of the present invention. Those skilled in the art will understand that the following examples are intended solely to illustrate the invention and should not be construed as limiting the scope of the invention. Any specific techniques or conditions not shown in the examples are carried out according to techniques or conditions described in the literature in the art or according to product manuals. Reagents or equipment not specified by the manufacturer are conventional products that are commercially available.

[0273] The amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein in Example 1 of the present invention is as shown in Sequence ID No. 1, and it should be noted that the fusion protein is prepared using conventional methods in the art.

[0274] Example 1 1. Examination of injectable solutions prepared using various buffer systems. 1.1 Injectable solutions of low-concentration formulations A1-A3 were prepared using various buffer systems. The final concentrations of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein, i.e., API) in the injectable solutions, as well as the types and final concentrations of excipients, are detailed in Table 1. The preparation method for the injectable solutions was as follows: Excipients and APIs were weighed and mixed according to the proportions listed in Table 1. If necessary, the pH of the formulation was adjusted to the corresponding value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the solution was adjusted to volume using sterile water for injection. After preparation, the drug solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 1 ml per syringe, and then sealed with rubber stoppers. Subsequently, formulations A1-A3 were subjected to high-temperature stability testing. The results of stability tests using SEC-HPLC after storage at 40°C for 0, 10, and 14 days are shown in Table 2.

[0275] [Table 1]

[0276] [Table 2]

[0277] Table 2 shows that the citrate buffer system exhibits the slowest rate of decline in RP-HPLC main peak purity, and visible particles can be observed to meet specifications after 14 days at 40°C. Therefore, with respect to low-concentration fusion protein formulations, the citrate buffer system exhibits relatively better chemical and physical stability.

[0278] Since various buffer systems may have different effects on the thermal stability of proteins, further investigations were conducted using differential scanning calorimetry (DSC) to determine the Tm and Tonset values ​​of formulations A1-A3 in various buffer systems. This investigation was conducted to investigate and compare the effects of various factors on the thermal stability of proteins based on temperature changes during protein denaturation. The DSC test results are shown in Table 3. Here, Tonset and Tm values ​​are indicators of thermal stability. The Tonset value represents the initial temperature at which protein unfolding begins, while the Tm value represents the intermediate temperature of the protein thermal transition. Generally, the higher the Tonset and Tm values, the more stable the protein is, and the less likely it is to unfold or denature in a low-temperature environment.

[0279] [Table 3]

[0280] Table 3 shows that, by comparing the Tonset, Tm1, and Tm2 values, the thermal stability of low-concentration fusion protein formulations in citrate buffer (formulation A1) and phosphate buffer (formulation A2) is superior to that in the histidine buffer system (formulation A3).

[0281] 1. Preparation of high-concentration injection solutions F1 - F4 using various buffer systems, the final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injection solution, as well as the type and final concentration of excipients, are detailed in Table 4. The preparation method for the injection solution was as follows: The fusion protein stock solution was ultrafiltered, concentrated, and then followed by buffer exchange. After measuring the protein concentration, the fusion protein solution was diluted to 25 mg / ml using the corresponding buffer system. If necessary, the pH was adjusted to 7.0 using 2% hydrochloric acid or 2% sodium hydroxide solution. After preparation, the drug solution was passed through 0.45 μm and 0.22 μm PVDF membranes in sequence and filtered into 2R borosilicate glass vials with a filling volume of 1 ml per vial. The vials were sealed with rubber stoppers and aluminum-plastic caps. Subsequently, formulations F1 - F4 were subjected to high-temperature stability investigation. The rate of change (% / day) of SEC-HPLC aggregates after storage at 40°C for 14 days is shown in Tables 5-1, 5-2, and 5-3.

[0282]

Table 4

[0283]

Table 5-1

[0284]

Table 5-2

[0285]

Table 5-3

[0286] Table 5-1 shows that after 14 days at 40°C, the growth rate of SEC aggregates was relatively fast in the sodium citrate buffer (formulation F1), phosphate buffer (formulation F3), and Tris buffer (formulation F4) systems. In contrast, histidine buffer (formulation F2) significantly reduced the growth rate of SEC aggregates (SEC aggregate reference rate is 5%). Regarding RP-HPLC total impurities (total impurities reference rate is 40%), the rate of change was similar across all four formulations. Therefore, it can be further concluded that formulations prepared with histidine buffer exhibit higher stability when the fusion protein concentration is higher than or equal to 25 mg / ml. However, all stability data for formulations using various buffer systems fall within acceptable limits.

[0287] 2. Examination of injection solutions prepared at various pH levels. 2.1 Low-concentration formulations A4-A8 were prepared using various pH values ​​for injection. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein, i.e., API) in the injection, as well as the types and final concentrations of the excipients, are detailed in Table 6. The preparation method for the injection was as follows: Excipients and API were weighed and mixed according to the proportions listed in Table 6. If necessary, the pH of the formulation was adjusted to the corresponding value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the solution was adjusted to volume using sterile water for injection. After preparation, the drug solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 1 ml per syringe, and then sealed with rubber stoppers. Subsequently, formulations A4-A8 were subjected to high-temperature stability studies and long-term stability studies. The results of stability tests using SEC-HPLC after storage at 40°C for 0, 10, and 14 days are shown in Table 7. The results of stability tests using SEC-HPLC after storage in a refrigerator at 4°C for 0, 1, 6, and 12 months are shown in Table 8.

[0288] [Table 6]

[0289] [Table 7]

[0290] [Table 8]

[0291] From Tables 7 and 8, it can be concluded that at pH 6.0, the injection solution exhibited the fastest decline in RP-HPLC main peak purity, the highest dimer content, and the poorest stability under high temperature conditions of 40°C. Additionally, after long-term storage for 12 months, visible particles did not meet specifications, SEC purity (monomers including shoulder peaks) declined the fastest, the dimer content was the highest, and stability was the poorest. In contrast, formulations at other pH levels showed good chemical stability.

[0292] Since various buffer systems may have different effects on the thermal stability of proteins, further investigations were conducted using differential scanning calorimetry (DSC) to determine the Tm and Tonset values ​​of formulations A4-A8 in various buffer systems. This investigation was conducted to investigate and compare the effects of various factors on the thermal stability of proteins based on temperature changes during protein denaturation. The DSC test results are shown in Table 9.

[0293] [Table 9]

[0294] From the DSC test results in Table 9, it can be observed that thermal stability is poorest at pH 6.0, while other pH groups exhibit relatively better stability.

[0295] In the range of 6.0 to 7.0, in order to further investigate the effect of pH on product stability, Formulations A9 - A15 were prepared at various pH levels. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injection solution, as well as the type and final concentration of the excipients, are detailed in Table 10. The preparation method for the injection solution follows the steps described in Section 2.1 of Example 1 for preparing Formulations A4 - A8. Subsequently, Formulations A9 - A15 were subjected to high-temperature stability investigation and DSC tests. The stability test results using SEC-HPLC after storage at 40°C for 0 days, 10 days, 14 days, and 21 days are shown in Table 11. The DSC test results are shown in Table 12.

[0296]

Table 10

[0297]

Table 11

[0298]

Table 12

[0299] From Tables 11 and 12, it can be observed that at pH values of 6.0 and 6.2, both the Tonset and Tm1 values are relatively low, indicating poor thermal stability compared to other groups. However, at pH values of 6.4 and above, all of the chemical stability, physical stability, and thermal stability of the injection solution are better.

[0300] 2.2 High-concentration injection solutions F5-F11 were prepared using various pH values. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein or API) in the injection solutions, as well as the types and final concentrations of excipients, are detailed in Table 13. The preparation method for the injection solutions was as follows: Excipients and APIs were weighed and mixed according to the formulation table. If necessary, the pH of the formulation was adjusted to the desired value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the mixture was diluted to a total volume of 10 ml using sterile water for injection. After preparation, the solutions were passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 0.5 ml per syringe, and then sealed with rubber stoppers. Subsequently, high-temperature stability studies were conducted on formulations F1-F4. Table 14 shows the rate of change (days / %) of SEC-HPLC aggregates after the formulation has been left at 40°C for 20 days.

[0301] [Table 13]

[0302] [Table 14]

[0303] Table 14 shows that after 20 days of storage at 40°C, within the pH range of 6.6–7.5, the increase in SEC aggregates slowed as the pH increased. At pH 6.6, high molecular weight proteins increased to 5.53% by day 20, exceeding the established quality standard limit of 5%. At pH values ​​of 6.7–7.5, the rate of SEC aggregate growth remained below 5%.

[0304] Furthermore, formulations F5 to F11 were subjected to a 72-hour room-temperature shaking experiment at a shaking speed of 600 rpm. The results of the shaking experiment are shown in Table 15 below.

[0305] [Table 15]

[0306] Table 15 shows that after 72 hours of shaking, formulations F5–F11 showed only a slight increase in SEC aggregates, and this increase slowed as the pH value increased. When the formulation pH was in the range of 6.6–7.5, this pH range effectively prevented the formation of excessive protein aggregates caused by shaking.

[0307] To further evaluate the effects of various pH values ​​on the three-dimensional and colloidal stability of proteins, formulations F5–F11 were tested using the NanoTemper Prometheus NT.48 protein stability analyzer with the ThermControl module. This analysis provided the protein denaturation temperature (Tm and Tonset) and the protein aggregation onset temperature (Tagg). The test results are shown in Table 16 below.

[0308] [Table 16]

[0309] Table 16 shows that at a pH of 6.6, the values ​​of Tonset, Tm, and Tagg were relatively low compared to other groups, indicating poorer stability. Between pH 6.7 and 7.5, Tonset and Tm1 values ​​increased with increasing pH, showing better thermal stability. Between pH 7.0 and 7.5, no aggregation temperature (Tagg) was detected, suggesting good colloidal stability.

[0310] In summary, the chemical and physical stability of a formulation is optimal when its pH is in the range of 6.7 to 7.5.

[0311] 3. Evaluation of injectable solutions prepared using various surfactants 3.1 The preparation of low-concentration injectable formulations A16-A19 using various surfactants, the final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein or API) in the injectable solutions, and the types and final concentrations of excipients are detailed in Table 17. The preparation method for the injectable solutions is as follows: Excipients and APIs were weighed and mixed according to the formulation table. If necessary, the pH of the formulation was adjusted to the desired value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the mixture was diluted to the specified volume using sterile water for injection. After preparation, the solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 0.8 ml per syringe, and then sealed with rubber stoppers. Formulations A16-A19 were subjected to high-temperature stability testing. The SEC-HPLC stability test results after 0 days and 10 days of storage at 40°C are shown in Table 18, and the SEC-HPLC stability test results after 10 days of storage at 40°C are shown in Table 19.

[0312] [Table 17]

[0313] [Table 18]

[0314] [Table 19]

[0315] From Tables 18 and 19, it can be concluded that various types of surfactants have a relatively small effect on SEC purity and aggregate formation, but a significant effect on RP-HPLC purity. Based on the analysis of the daily decrease rate of RP-HPLC purity (main peak + peak 4) and the daily increase rate of RP-HPLC purity (total related substances), polysorbate 80 or polysorbate 20 show excellent performance as stabilizers. However, when poloxamer 188 is used as a stabilizer, the formulation stability remains within acceptable limits.

[0316] 3.2 Preparation of high-concentration injectable formulations F12 and F13 using various surfactants. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein or API) in the injectable solution, as well as the type and final concentration of excipients, are detailed in Table 20. The preparation method for the injectable solution is as follows: Excipients and API were weighed and mixed according to the formulation table. If necessary, the pH of the solution was adjusted to the desired value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the mixture was diluted to a total volume of 15 ml with sterile water for injection. After preparation, the solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 0.5 ml per syringe, and then sealed with rubber stoppers. Formulations F12 and F13 were subjected to high-temperature stability testing. The rate of change (days / %) of SEC-HPLC aggregates after 28 days of storage at 40°C is shown in Table 21, and the visual inspection results are shown in Table 22.

[0317] [Table 20]

[0318] [Table 21]

[0319] [Table 22]

[0320] The data from Tables 21 and 22 demonstrate that formulations prepared with polysorbate 80 and polysorbate 20 effectively slow the growth of SEC aggregates, prevent the formation of visible particles, and provide excellent stabilization effects on fusion proteins.

[0321] 4. Evaluation of injectable solutions prepared using various osmotic regulators. 4.1 Low-concentration injection solutions A23-A25 were prepared using various osmotic regulators. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injection solutions, as well as the types and final concentrations of excipients, are detailed in Table 23. The preparation methods for the injection solutions are described in step 2.1 of the experimental procedure in this application. Formulations A23-A25 were subjected to high-temperature stability testing. The results of SEC-HPLC stability tests after storage at 40°C for 0, 7, and 14 days are shown in Table 24.

[0322] [Table 23]

[0323] [Table 24]

[0324] Table 24 shows that, compared to formulations using sodium chloride or a combination of sodium chloride and sucrose, formulations using mannitol as an osmotic regulator exhibit the slowest growth rate of RP-HPLC purity (total related substances) and fewer visible particles. Therefore, formulations prepared using mannitol as an osmotic regulator exhibit superior stability.

[0325] Additionally, the osmotic pressure of low-concentration injectable solutions prepared with mannitol was measured and found to be equal to or close to that of human plasma osmolality. The results are as follows: Formulation A12: 307 mOsm / kg Formulation A16: 310 mOsm / kg Formulation A26: 309 mOsm / kg Formulation A29: 325 mOsm / kg.

[0326] 4.2 Preparation of low-concentration injectable formulations A34-A35 using various osmotic regulators. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injectable formulations, as well as the type and final concentration of excipients, are detailed in Table 25-1. The preparation method for the injectable formulations is described in step 2.1 of the experimental procedure in this application. Formulations A34-A35 were subjected to high-temperature stability testing. The results of SEC-HPLC stability tests after storage at 40°C for 0, 7, and 14 days are shown in Table 25-2.

[0327] [Table 25-1]

[0328] [Table 25-2]

[0329] Formulations prepared using sucrose or trehalose as osmotic regulators also exhibit excellent stability.

[0330] 4.3 Preparation of high-concentration injectable formulations F14 and F15 using various osmotic regulators. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein or API) in the injectable solution, as well as the type and final concentration of excipients, are detailed in Table 26. The preparation method for the injectable solution is as follows: Excipients and API were weighed and mixed according to the formulation table. If necessary, the pH of the solution was adjusted to the desired value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the mixture was diluted to a total volume of 9 ml with sterile water for injection. After preparation, the solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 0.5 ml per syringe, and then sealed with rubber stoppers. Formulations F14 and F15 were subjected to high-temperature stability testing. The rate of change of SEC-HPLC aggregates after 30 days of storage at 40°C (days / %) is shown in Table 27, and the visual inspection results after 14 days of storage at 40°C are shown in Table 28.

[0331] [Table 26]

[0332] [Table 27]

[0333] [Table 28]

[0334] To further evaluate the effects of various osmotic regulators on the three-dimensional and colloidal stability of proteins, formulations F14–F15 were tested using the ThermControl module of a NanoTemper Prometheus NT.48 protein stability analyzer. This analysis provided the protein denaturation temperatures (Tm and Tonset) and the protein aggregation onset temperature (Tagg). The test results are shown in Table 29 below.

[0335] [Table 29]

[0336] From Tables 27-29, it can be concluded that both mannitol and sucrose enable the dual-target fusion protein formulation to maintain good colloidal and conformational stability. Mannitol and sucrose also slow the rate of protein aggregation, prevent the formation of visible particles, and provide excellent stabilization effects on the fusion protein.

[0337] 5. Evaluation of injectable solutions prepared using various histidine concentrations. Preparation of high-concentration injectable formulations F16-F18 using various histidine concentrations. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein or API) in the injectable solution, as well as the type and final concentration of excipients, are detailed in Table 30. The preparation method for the injectable solution is as follows: Excipients and API were weighed and mixed according to the formulation table. If necessary, the pH of the solution was adjusted to the desired value using 2% hydrochloric acid solution or 2% sodium hydroxide solution. Finally, the mixture was diluted to a total volume of 10 ml with sterile water for injection. After preparation, the solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 0.5 ml per syringe, and then sealed with rubber stoppers. Formulations F16-F18 were subjected to high-temperature stability testing. The rate of change (days / %) of SEC-HPLC aggregates after 30 days of storage at 40°C is shown in Table 31.

[0338] [Table 30]

[0339] [Table 31]

[0340] From the above, it can be seen that when the histidine concentration is 10-20 mM, the growth rate of SEC aggregates in the fusion protein is relatively slow, indicating that histidine has a good stabilizing effect on the fusion protein.

[0341] 6. Examination of injectable solutions prepared using various polysorbate 80 concentrations. 6.1 Injectable solutions with low concentrations (A20-A22 and A32-A33) were prepared using polysorbate 80 at various concentrations. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injectable solutions, as well as the types and final concentrations of excipients, are detailed in Table 32. The preparation method for the injectable solutions is described in step 2.1 of the examples in this application. Formulations A20-A22 and A32-A33 were then subjected to high-temperature stability tests. The results of SEC-HPLC stability tests after storage at 40°C for 0, 7, 14, and 28 days are shown in Table 33.

[0342] [Table 32]

[0343] [Table 33]

[0344] Table 33 shows that the stability of the injection solution is optimal when the dosage of polysorbate 80 is within the range of 0.1 mg / ml to 0.5 mg / ml.

[0345] 6.2 High-concentration injectable formulations F19-F25 were prepared using polysorbate 80 at various concentrations. The final concentrations of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein, i.e., API) in the injectable solutions, as well as the types and final concentrations of excipients, are detailed in Table 34. The preparation method for the injectable solutions was as follows: Excipients and APIs were weighed and mixed according to the table below. If necessary, the pH of the solution was adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution. Finally, the mixture was diluted to a total volume of 15 ml with sterile water for injection. After preparation, the solutions were successively passed through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 0.5 ml per syringe, and then sealed with rubber stoppers. Formulations F19-F25 were then subjected to high-temperature stability testing. The SEC-HPLC aggregate growth rate (% / day) after 28 days of storage at 40°C is shown in Table 35, the visual inspection results are shown in Table 36, and the insoluble particle test results are shown in Table 37.

[0346] [Table 34]

[0347] [Table 35]

[0348] [Table 36]

[0349] [Table 37]

[0350] From the above, it can be concluded that polysorbate 80 can maintain the stability of dual-target fusion proteins when its concentration is within the range of 0.4 to 1.3 mg / ml. The growth rate of protein aggregates is relatively slow, and the number of insoluble particles is low, indicating that polysorbate 80 prevents the formation of visible particles and reduces the formation of insoluble particles.

[0351] 7. Examination of injection solutions prepared using various fusion protein concentrations. 7.1 Preparation of low-concentration injectable formulations A26-A31 using various fusion protein concentrations. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injectable formulations, as well as the types and final concentrations of excipients, are detailed in Table 38. The preparation method for the injectable formulations is described in step 2.1 of the embodiments of this application. Subsequently, formulations A26-A31 were subjected to high-temperature stability testing. The results of SEC-HPLC stability tests after storage at 40°C for 0, 7, 14, and 28 days are shown in Table 39. The aggregate growth trends for formulations of various concentrations under high-temperature SEC conditions are illustrated in Figure 1, and the growth trends for purity (total related substances) under high-temperature RP-HPLC conditions are shown in Figure 2.

[0352] [Table 38]

[0353] [Table 39]

[0354] From Table 39 and Figures 1-2, it can be observed that the growth rate of RP-HPLC purity (total related substance) does not show a significant difference among formulations of various concentrations. However, the growth rate (%) of SEC purity (aggregates) increases with higher formulation concentrations.

[0355] Furthermore, the long-term stability of formulations A29 (17 mg / ml) and A30 (20 mg / ml) at 2–8°C was investigated. The test results are shown in Table 40.

[0356] [Table 40]

[0357] Table 40 shows that the RP-HPLC purity (main peak + peak 4) of formulations at various concentrations did not show significant changes over the storage period. SEC purity (aggregates) showed a slight increasing trend but remained within the acceptable range (≤5%).

[0358] 7.2 Preparation of low-concentration injectable formulations F26-F29 using various fusion protein concentrations. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injectable formulations, as well as the types and final concentrations of excipients, are detailed in Table 41. The preparation method for the injectable formulations is described in step 2.1 of the embodiments of this application. Subsequently, formulations F26-F29 were subjected to high-temperature stability testing. The SEC-HPLC stability test results after storage at 40°C for 0, 7, 14, and 28 days showed a slight increasing trend in SEC purity (aggregates) of formulations with various concentrations, but remained within an acceptable range (≤5%). This embodiment exemplifies the results for formulation F26, as detailed in Tables 42 and 43.

[0359] [Table 41]

[0360] [Table 42]

[0361] [Table 43]

[0362] 8. Effect of EDTA sodium salt on formulation stability in various buffer systems 8.1 Preparation of low-concentration injectable formulations A36-A37 containing EDTA in a citrate buffer system. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injectable solution, as well as the type and final concentration of excipients, are detailed in Table 44. The preparation method for the injectable solution is described in step 2.1 of the embodiments of this application. Subsequently, the high-temperature stability of formulations A36-A37 was investigated. The results of SEC-HPLC stability tests after storage at 40°C for 0, 10, 14, and 28 days are shown in Table 45.

[0363] [Table 44]

[0364] [Table 45]

[0365] 8.2 Preparation of high-concentration injectable formulations F30-F31 in histidine-based formulations containing EDTA. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injectable formulations, as well as the type and final concentration of excipients, are detailed in Table 46. The preparation method for the injectable formulations is described in step 2.1 of the examples in this application. Subsequently, high-temperature stability tests were performed on formulations F30-F31. The results of SEC-HPLC stability tests after storage at 40°C for 0, 7, 14, and 30 days are shown in Table 47.

[0366] [Table 46]

[0367] [Table 47]

[0368] In summary, the formulations exhibited excellent stability regardless of whether they were citrate-based or histidine buffer-based, and regardless of whether EDTA sodium salt was added.

[0369] Example 2: Evaluation of various fusion proteins The amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 2 in this example is shown in SEQ ID NO: 2; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 3 in this example is shown in SEQ ID NO: 3; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 4 in this example is shown in SEQ ID NO: 4; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 5 in this example is shown in SEQ ID NO: 5; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 6 in this example is shown in SEQ ID NO: 6; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein The amino acid sequence of protein 7 is shown in SEQ ID NO: 7; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 8 in this example is shown in SEQ ID NO: 8; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 9 in this example is shown in SEQ ID NO: 9; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 10 in this example is shown in SEQ ID NO: 10; the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 11 in this example is shown in SEQ ID NO: 11; and the amino acid sequence of GLP-1-Fc-FGF21 dual target fusion protein 12 in this example is shown in SEQ ID NO: 12. Of these GLP-1-Fc-FGF21 dual target fusion proteins, GLP-1-Fc-FGF21 dual target fusion proteins 2 to 11 (referred to as fusion proteins 2 to 11) were prepared using conventional methods in this field.

[0370] The type and final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injection solution, as well as the type and final concentration of the excipients, are detailed in Table 48. The preparation method for the injection solution is described in step 2.1 of Example 1. Subsequently, stability tests were performed on formulations A38 to A31.

[0371] [Table 48]

[0372] 2. The type and final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein) in the injection solution, as well as the type and final concentration of the excipients, are detailed in Table 49. The preparation method for the injection solution is described in step 2.1 of Example 1. Subsequently, stability tests were performed on formulations F32 to F53.

[0373] [Table 49]

[0374] Example 3: Evaluation of various preservative systems Low-concentration injectable formulations were prepared using various preservative systems. The final concentrations of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as the fusion protein or API) in the injectable solutions, as well as the types and final concentrations of excipients, are detailed in Table 50. The preparation method for the injectable solutions was as follows: Excipients and APIs were weighed and mixed according to the proportions listed in Table 1. If necessary, the pH of the formulation was adjusted to the corresponding value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution. Finally, the solution was adjusted to volume using sterile water for injection. After preparation, the drug solution was passed sequentially through 0.45 μm and 0.22 μm PVDF membranes and filtered into 1 ml elongated pre-filled syringes with a filling volume of 1 ml per syringe, and then sealed with rubber stoppers. Subsequently, stability tests were performed on the formulations. The results showed that all formulations exhibited good stability, with an acceptable rate of SEC-HPLC aggregate change (days / %) within acceptable limits, and no visible particles were formed.

[0375] [Table 50]

[0376] In this specification, references to terms such as “one embodiment,” “some embodiments,” “examples,” “specific examples,” or “some examples” mean that the specific characteristics, structures, materials, or features described in relation to an embodiment or example are included in at least one embodiment or example of the present invention. Therefore, the appearance of phrases such as “in some embodiments,” “in one embodiment,” “in one embodiment,” “in another embodiment,” “in one embodiment,” “specific examples,” or “in some embodiments” in various places throughout this specification does not necessarily refer to the same embodiment or example of this disclosure. Furthermore, specific characteristics, structures, materials, or features can be combined in any suitable manner in one or more embodiments or examples. In addition, those skilled in the art can integrate and combine different embodiments, examples, or their characteristics, provided that the embodiments, examples, or their characteristics are not inconsistent with each other.

[0377] Although explanatory embodiments are shown and described, those skilled in the art will understand that the above embodiments are not to be construed as limiting the disclosure, and that variations, substitutions, and modifications may be made in embodiments without departing from the spirit, principles, and scope of the disclosure.

Claims

1. A composition comprising a GLP-1-Fc-FGF21 fusion protein and a buffer.

2. The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequences shown in SEQ ID NOs: 1 to 12; Optionally, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 1 to 100 mg / mL; Optionally, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 80 mg / mL; Optionally, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 5 to 70 mg / mL; Optionally, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 55 mg / mL; Optionally, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 10 to 65 mg / mL; Optionally, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 20 to 60 mg / mL; Optionally, the pH value of the composition is 6.2 to 8.0, preferably 6.4 to 8.

0. The composition according to claim 1.

3. The buffer solution comprises at least one of histidine buffer, citrate buffer, phosphate buffer, and Tris buffer; Preferably, the buffer solution comprises a histidine buffer or a citrate buffer; Optionally, the concentration of the buffer in the composition is 5 to 30 mM; Optionally, the concentration of the buffer in the composition is 5 to 25 mM; Optionally, the concentration of the buffer in the composition is 8 to 25 mM; Optionally, the concentration of the buffer in the composition is 5 to 20 mM; Optionally, the concentration of the buffer in the composition is 8 to 12 mM; Preferably, the concentration of the buffer solution in the composition is 10 to 20 mM. The composition according to claim 2.

4. The composition further comprises at least one of an osmotic pressure regulator, a surfactant, a preservative, and a metal chelating agent; Optionally, the osmotic pressure regulator comprises at least one of mannitol, sucrose, trehalose, and sodium chloride; Optionally, the concentration of the osmotic pressure regulator in the composition is 30 to 100 mg / mL or 1 to 10 mg / mL; Optionally, the surfactant comprises at least one of polysorbate 80, polysorbate 20, and poloxamer 188; Optionally, the concentration of the surfactant in the composition is 0.1 to 10 mg / mL, preferably 0.1 to 1.5 mg / mL or 6 to 10 mg / mL, more preferably 0.1 to 0.7 mg / mL; The preservative optionally comprises at least one of phenol and m-cresol; Optionally, the concentration of the preservative in the composition is 0.1 to 10 mg / mL, preferably 1.0 to 5.0 mg / mL; The metal chelating agent optionally comprises at least one of disodium edetate and calcium disodium edetate; Optionally, the concentration of the metal chelating agent in the composition is 0.01 to 1 mg / mL, preferably 0.05 to 0.5 mg / mL. The composition according to claim 1.

5. The buffer is a histidine buffer, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, or 25 to 85 mg / mL; Optionally, the buffer is a histidine buffer, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25 to 80 mg / mL, preferably 25 to 55 mg / mL; Optionally, the concentration of the histidine buffer in the composition is 5 to 30 mM, preferably 5 to 25 mM, 8 to 25 mM, or 8 to 20 mM, more preferably 10 to 20 mM or 8 to 12 mM; Optionally, the pH of the composition is 6.7 to 7.5, preferably 6.9 to 7.5; Optionally, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30 to 50 mg / mL; or, The osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60 to 100 mg / mL, preferably 80 to 100 mg / mL; or, The osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40 to 100 mg / mL, preferably 40 to 70 mg / mL, and more preferably 50 to 70 mg / mL; Optionally, the surfactant is polysorbate 80, and the concentration of the surfactant in the composition is 0.1 to 2 mg / mL, preferably 0.1 to 1.5 mg / mL, more preferably 0.4 to 1.3 mg / mL; or, The surfactant is polysorbate 20, and the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL, preferably 0.1 to 0.5 mg / mL. The composition according to any one of claims 1 to 4.

6. The concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2 to 25 mg / mL, and the buffer is a citrate buffer; Optionally, the concentration of the citrate buffer in the composition is 5 to 30 mM, preferably 5 to 25 mM, 8 to 25 mM, or 8 to 20 mM, more preferably 10 to 20 mM or 8 to 12 mM; Optionally, the pH of the composition is 6.4 to 8.0; Optionally, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30 to 50 mg / mL; or, The osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60 to 100 mg / mL, preferably 80 to 100 mg / mL; or, The osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40 to 100 mg / mL, preferably 40 to 70 mg / mL, more preferably 50 to 70 mg / mL; The osmotic pressure regulator is sodium chloride, and the concentration of the osmotic pressure regulator in the composition is 1 to 10 mg / mL, preferably 3 to 9 mg / mL; Optionally, the surfactant comprises at least one of polysorbate 80, polysorbate 20, and poloxamer 188, preferably polysorbate 80; Optionally, the concentration of the surfactant in the composition is 0.1 to 10 mg / mL, preferably 0.1 to 1.5 mg / mL, 0.1 to 1.0 mg / mL, or 6 to 10 mg / mL; Optionally, the surfactant is poloxamer 188, and the concentration of the surfactant in the composition is 6 to 10 mg / mL; or, The surfactant is polysorbate 80 or polysorbate 20, and the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL, preferably 0.1 to 0.5 mg / mL. The composition according to any one of claims 1 to 4.

7. The composition according to any one of claims 1 to 4, wherein the dosage form of the composition is a liquid formulation, preferably an injectable solution.

8. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, 25 to 70 mg / mL, 25 to 65 mg / mL, 25 to 60 mg / mL, 25 to 55 mg / mL, or 25 to 50 mg / mL; The concentration of the histidine buffer in the injection solution is 8 to 25 mM, preferably 10 to 20 mM; The concentration of mannitol in the aforementioned injection solution is 35 to 45 mg / mL; The concentration of polysorbate 80 in the injection solution is 0.4 to 1.5 mg / mL or 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1 to 0.5 mg / mL; The concentration of the metal chelating agent in the aforementioned injection solution is 0.05 to 0.5 mg / mL; The pH of the aforementioned injection solution is 6.9 to 7.5; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

9. GLP-1-Fc-FGF21 fusion protein, Histidine buffer, Sucrose or trehalose, Polysorbate 80 or polysorbate 20, and Optionally, choose either disodium edetate or calcium disodium edetate. An injectable solution containing; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, 25 to 70 mg / mL, 25 to 65 mg / mL, 25 to 60 mg / mL, 25 to 55 mg / mL, or 25 to 50 mg / mL; The concentration of the histidine buffer in the injection solution is 8 to 25 mM, preferably 10 to 20 mM; The concentration of sucrose in the aforementioned injection solution is 85 to 95 mg / mL; The concentration of trehalose in the aforementioned injection solution is 55 to 65 mg / mL; The concentration of polysorbate 80 in the injection solution is 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1 to 0.5 mg / mL; The pH of the aforementioned injection solution is 6.9 to 7.5; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

10. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, citrate buffer, sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, 25 to 70 mg / mL, 25 to 65 mg / mL, 25 to 60 mg / mL, 25 to 55 mg / mL, or 25 to 50 mg / mL; The concentration of the citrate buffer in the injection solution is 8 to 25 mM, preferably 10 to 20 mM; The concentration of sucrose in the aforementioned injection solution is 85 to 95 mg / mL; The concentration of trehalose in the aforementioned injection solution is 55 to 65 mg / mL; The concentration of polysorbate 80 in the injection solution is 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1 to 0.5 mg / mL; The pH of the aforementioned injection solution is 6.9 to 7.5; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

11. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and optionally disodium edetate or calcium disodium edetate, and polysorbate 80 or polysorbate 20; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 100 mg / mL, preferably 25 to 95 mg / mL, 25 to 90 mg / mL, 25 to 85 mg / mL, 25 to 80 mg / mL, 25 to 75 mg / mL, 25 to 70 mg / mL, 25 to 65 mg / mL, 25 to 60 mg / mL, 25 to 55 mg / mL, or 25 to 50 mg / mL; The concentration of the citrate buffer in the injection solution is 8 to 25 mM, preferably 10 to 20 mM; The concentration of mannitol in the aforementioned injection solution is 35 to 45 mg / mL; The concentration of polysorbate 80 in the injection solution is 0.4 to 1.5 mg / mL or 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the injection solution is 0.1 to 0.5 mg / mL; The concentration of the metal chelating agent in the aforementioned injection solution is 0.05 to 0.5 mg / mL; The pH of the aforementioned injection solution is 6.9 to 7.5; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

12. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium disodium edetate; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; The concentration of the citrate buffer in the injection solution is 5 to 20 mM, preferably 8 to 20 mM, more preferably 10 to 15 mM; The concentration of mannitol in the aforementioned injection solution is 40-50 mg / mL; The concentration of polysorbate 80 in the aforementioned injection solution is 0.1 to 1.5 mg / mL; The concentration of the metal chelating agent in the aforementioned injection solution is 0.05 to 0.5 mg / mL; The pH of the aforementioned injection solution is 6.4 to 8.0; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

13. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80, and sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; The concentration of the citrate buffer in the injection solution is 5 to 20 mM, preferably 8 to 20 mM, more preferably 10 to 15 mM; The concentration of sucrose in the aforementioned injection solution is 85 to 95 mg / mL; The concentration of trehalose in the aforementioned injection solution is 55 to 65 mg / mL; The concentration of polysorbate 80 in the aforementioned injection solution is 0.1 to 1.5 mg / mL; The pH of the aforementioned injection solution is 6.4 to 8.0; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

14. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, polysorbate 80, and optionally disodium edetate or calcium disodium edetate; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; The concentration of the histidine buffer in the injection solution is 5 to 30 mM, preferably 10 to 20 mM; The concentration of mannitol in the aforementioned injection solution is 40-50 mg / mL; The concentration of polysorbate 80 in the aforementioned injection solution is 0.1 to 1.5 mg / mL; The concentration of the metal chelating agent in the aforementioned injection solution is 0.05 to 0.5 mg / mL; The pH of the aforementioned injection solution is 6.4 to 8.0; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

15. An injectable solution comprising GLP-1-Fc-FGF21 fusion protein, histidine buffer, polysorbate 80, and sucrose or trehalose, and optionally disodium edetate or calcium disodium edetate; The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in any one of SEQ ID NOs: 1 to 12, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 3 to 25 mg / mL; The concentration of the histidine buffer in the injection solution is 5 to 30 mM, preferably 10 to 20 mM; The concentration of sucrose in the aforementioned injection solution is 85 to 95 mg / mL; The concentration of trehalose in the aforementioned injection solution is 55 to 65 mg / mL; The concentration of polysorbate 80 in the aforementioned injection solution is 0.1 to 1.5 mg / mL; The pH of the aforementioned injection solution is 6.4 to 8.0; Preferably, the injection solution does not contain disodium edetate or calcium disodium edetate. injection solution.

16. The injectable solution according to any one of claims 8 to 15, further comprising a preservative.

17. The preservative is at least one selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol; Optionally, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, preferably 3 to 7 mg / mL; Optionally, the concentration of m-cresol in the composition or injection solution is 0 to 4 mg / mL, preferably 2 to 4 mg / mL. The injection solution according to claim 16.

18. The use of compositions or injectable solutions in the preparation of pharmaceuticals for the treatment and / or prevention of metabolic diseases; Optionally, the metabolic diseases include diseases associated with diabetes mellitus, dyslipidemia, and fatty liver disease; Optionally, the diseases associated with fatty liver include non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis. Use of the composition according to any one of claims 1 to 7 or the injection solution according to any one of claims 8 to 17.

19. A method for treating and / or preventing metabolic diseases, comprising the step of administering a composition according to any one of claims 1 to 7 or an injectable solution according to any one of claims 8 to 17 to a subject, in the preparation of a medicament for treating and / or preventing metabolic diseases; Optionally, the metabolic diseases include diseases associated with diabetes mellitus, dyslipidemia, and fatty liver disease; Optionally, the diseases associated with fatty liver include non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis. method.

20. A composition or injection for use in treating and / or preventing metabolic diseases; Optionally, the metabolic diseases include diseases associated with diabetes mellitus, dyslipidemia, and fatty liver disease; Optionally, the diseases associated with fatty liver include non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis. The composition according to any one of claims 1 to 7, or the injectable solution according to any one of claims 8 to 17.

21. An injection device or container comprising the composition according to any one of claims 1 to 7 or the injection solution according to any one of claims 8 to 17; Optionally, the injection device or container includes at least one selected from bottles, vials, cartridges, and syringes; Preferably, the syringe includes at least one selected from an auto-injector and a pre-filled syringe. Injection device or container.

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