GLP-1-Fc-FGF21 double-target fusion protein composition and injection and application thereof

Abstract

The invention relates to a GLP-1-Fc-FGF21 double-target fusion protein composition and an application thereof. The composition is prepared from the GLP-1-Fc-FGF21 fusion protein and a buffer solution. According to the composition disclosed by the invention, by adding the buffer solution, the generation of visible foreign matters can be effectively controlled, and the stability of the GLP-1-Fc-FGF21 fusion protein can be improved, so that the shelf life of the product can be prolonged, and the composition disclosed by the invention can be used for treating and / or preventing metabolic diseases, such as diabetes mellitus, hyperlipidemia, obesity and fatty liver related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of biopharmaceutical technology. Specifically, the present invention relates to a GLP-1-Fc-FGF21 dual-target fusion protein composition and injection and uses thereof. More specifically, the present invention relates to a composition and injection and uses thereof and an injection device or container. Background Art

[0002] Glucagon-like peptide-1 (GLP-1) is an incretin hormone secreted by the L cells of the small intestine, which can stimulate the pancreatic β cells to secrete insulin, thereby maintaining the balance of insulin in the patient's body. GLP-1 works indirectly through insulin and only works on type 2 diabetes, which limits its scope of application and effect; at the same time, there are reports that GLP-1 has a potential risk of causing thyroid cancer.

[0003] FGF21 is a member of the FGF family (fibroblast growth factor, FGFs). FGF21 can promote the absorption of glucose by fat cells and enhance insulin sensitivity. Compared with insulin, FGF21 does not cause side effects such as hypoglycemia, and can more effectively protect pancreatic β cells, promote the regeneration and repair of pancreatic β cells. Moreover, since it has no mitogenic activity, it will not lead to potential tumor risks. FGF21 has the potential to become a drug for the treatment of type 1 diabetes. In addition, FGF21 also has a good lipid-lowering effect and is a very potential lipid-lowering drug.

[0004] GLP-1-Fc-FGF21 dual-target fusion protein can be used to treat metabolic diseases, such as diabetes, hyperlipidemia, obesity, and fatty liver-related diseases. However, the dual-target fusion protein will undergo physical and chemical changes during storage, such as denaturation, aggregation, precipitation, or fragmentation, which will affect the stability of the dual-target fusion protein during storage.

[0005] Therefore, there is an urgent need to develop a GLP-1-Fc-FGF21 dual-target fusion protein composition with high stability. Summary of the invention

[0006] The present invention aims to solve at least one of the technical problems existing in the prior art to a certain extent. To this end, the present invention provides a GLP-1-Fc-FGF21 dual-target fusion protein composition, which has strong stability and can extend the shelf life of the product.

[0007] In one aspect of the present invention, the present invention provides a composition. According to an embodiment of the present invention, the composition comprises: a GLP-1-Fc-FGF21 fusion protein and a buffer. In the composition of the present invention, by adding a buffer, the generation of visible foreign matter and aggregates can be effectively controlled, the stability of the GLP-1-Fc-FGF21 fusion protein can be improved, and thus the shelf life of the product can be extended.

[0008] According to an embodiment of the present invention, the GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence shown in SEQ ID NOs: 1-12.

[0009] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 1 to 100 mg / mL.

[0010] 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.

[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 70 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 5 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 2 to 65 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 60 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 55 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 10-65 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 20-60 mg / mL.

[0018] According to an embodiment of the present invention, the pH value of the composition is 6.2-8.0, preferably 6.4-8.0.

[0019] According to an embodiment of the present invention, the buffer comprises at least one of a histidine buffer, a citrate buffer, a phosphate buffer and a Tris buffer.

[0020] According to an embodiment of the present invention, the buffer comprises histidine buffer or citrate buffer.

[0021] According to an embodiment of the present invention, the concentration of the buffer in the composition is 5-30 mM.

[0022] According to an embodiment of the present invention, the concentration of the buffer in the composition is 5-25 mM.

[0023] According to an embodiment of the present invention, the concentration of the buffer in the composition is 8-25 mM.

[0024] According to an embodiment of the present invention, the concentration of the buffer in the composition is 5-20 mM.

[0025] According to an embodiment of the present invention, the concentration of the buffer in the composition is 8-12 mM.

[0026] According to an embodiment of the present invention, the concentration of the buffer in the composition is 10-20 mM.

[0027] According to an embodiment of the present invention, the composition further includes at least one of an osmotic pressure regulator, a surfactant, a preservative and a metal chelator.

[0028] According to an embodiment of the present invention, the osmotic pressure regulator includes at least one of mannitol, sucrose, sodium chloride and trehalose.

[0029] According to an embodiment of the present invention, the concentration of the osmotic pressure regulator in the composition is 30-100 mg / mL or 1-10 mg / mL.

[0030] According to an embodiment of the present invention, the surfactant includes at least one of polysorbate 80, polysorbate 20 and poloxamer 188.

[0031] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-10 mg / mL.

[0032] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-1.5 mg / mL.

[0033] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 6-10 mg / mL.

[0034] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-0.7 mg / mL.

[0035] According to an embodiment of the present invention, the preservative is at least one of phenol and m-cresol.

[0036] According to an embodiment of the present invention, the concentration of the preservative in the composition is 0.1-10 mg / mL, preferably 0.1-5.0 mg / mL, more preferably 1.0-5.0 mg / mL.

[0037] According to an embodiment of the present invention, the metal chelator is at least one of edetate disodium and edetate calcium sodium.

[0038] According to an embodiment of the present invention, the concentration of the metal chelator in the composition is 0.01 to 1 mg / mL, preferably 0.05 to 0.5 mg / mL.

[0039] According to an 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 25-100 mg / mL, preferably 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL.

[0040] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 25-80 mg / mL, and the buffer is a histidine buffer.

[0041] According to an embodiment 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 a histidine buffer.

[0042] According to an embodiment of the present invention, the concentration of the histidine buffer in the composition is 5-30 mM.

[0043] According to an embodiment of the present invention, the concentration of the histidine buffer in the composition is 5-25 mM.

[0044] According to an embodiment of the present invention, the concentration of the histidine buffer in the composition is 8-25 mM.

[0045] According to an embodiment of the present invention, the concentration of the histidine buffer in the composition is 8-20 mM.

[0046] According to an embodiment of the present invention, the concentration of the histidine buffer in the composition is 10-20 mM.

[0047] According to an embodiment of the present invention, the concentration of the histidine buffer in the composition is 8-12 mM.

[0048] According to an embodiment of the present invention, the pH value of the composition is 6.7-7.5, preferably 6.9-7.5.

[0049] According to an embodiment of the present invention, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30-50 mg / mL; or the osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60-100 mg / mL, preferably 80-100 mg / mL; or the osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40-100 mg / mL, preferably 40-70 mg / mL, and more preferably 50-70 mg / mL.

[0050] According to an 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, and 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, and preferably 0.1 to 0.5 mg / mL.

[0051] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2-25 mg / mL, and the buffer is a citrate buffer.

[0052] According to an embodiment of the present invention, the concentration of the citrate buffer in the composition is 5-30 mM.

[0053] According to an embodiment of the present invention, the concentration of the citrate buffer in the composition is 5-25 mM.

[0054] According to an embodiment of the present invention, the concentration of the citrate buffer in the composition is 8-25 mM.

[0055] According to an embodiment of the present invention, the concentration of the citrate buffer in the composition is 8-20 mM.

[0056] According to an embodiment of the present invention, the concentration of the citrate buffer in the composition is 10-20 mM.

[0057] According to an embodiment of the present invention, the concentration of the citrate buffer in the composition is 8-12 mM.

[0058] According to an embodiment of the present invention, the pH value of the composition is 6.4-8.0.

[0059] According to an embodiment of the present invention, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30-50 mg / mL; or the osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60-100 mg / mL, preferably 80-100 mg / mL; or the osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40-100 mg / mL, preferably 40-70 mg / mL, and more preferably 50-70 mg / mL.

[0060] According to an 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.

[0061] According to an embodiment of the present invention, the surfactant includes at least one of polysorbate 80, polysorbate 20 and poloxamer 188, preferably polysorbate 80.

[0062] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-10.0 mg / mL.

[0063] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-1.5 mg / mL.

[0064] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-1.0 mg / mL.

[0065] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 6.0-10.0 mg / mL.

[0066] According to an embodiment of the present invention, the concentration of the surfactant in the composition is 0.1-0.5 mg / mL.

[0067] According to an 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.

[0068] According to an embodiment of the present invention, the dosage form of the composition is an injection.

[0069] In another aspect of the present invention, the present invention provides an injection solution. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as 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 is 25 to 100 mg / mL; the concentration of the histidine buffer in the injection is 8 to 25 mM; the concentration of mannitol in the injection is 35 to 45 mg / mL; the concentration of polysorbate 80 in the injection is 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the injection is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0070] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0071] In another aspect of the present invention, the present invention provides an injection. According to an embodiment of the present invention, the injection comprises: a GLP-1-Fc-FGF21 fusion protein, a histidine buffer, mannitol and optionally edetate disodium or edetate calcium sodium, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has a SEQ ID NO: The amino acid sequence shown in any one of items 1 to 12, wherein the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 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 composition is 8 to 25 mM; the concentration of the mannitol in the composition is 35 to 45 mg / mL; the concentration of the polysorbate 80 in the composition is 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the composition is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the composition is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0072] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0073] In another aspect of the present invention, the present invention provides an injection solution. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25 to 50 mg / mL; the concentration of the histidine buffer in the injection is 10 to 20 mM; the concentration of mannitol in the injection is 35 to 45 mg / mL; the concentration of polysorbate 80 in the injection is 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the injection is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0074] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0075] In another aspect of the present invention, the present invention provides an injection solution. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose, and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 is 25 to 100 mg / mL; the concentration of the histidine buffer in the injection is 8 to 25 mM; the concentration of the sucrose in the injection is 85 to 95 mg / mL or the concentration of the trehalose in the injection is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection is 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0076] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0077] In another aspect of the present invention, the present invention provides an injection. According to an embodiment of the present invention, the injection comprises: a GLP-1-Fc-FGF21 fusion protein, a histidine buffer, sucrose or trehalose, and optionally edetate disodium or edetate calcium sodium, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has a SEQ ID NO: The amino acid sequence shown in any one of items 1 to 12, wherein the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 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 composition is 8 to 20 mM; the concentration of the sucrose in the composition is 85 to 95 mg / mL; the concentration of the trehalose in the injection is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the composition is 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the composition is 0.1 to 0.5 mg / mL; and the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0078] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0079] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: 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, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25-50 mg / mL; the concentration of the histidine buffer in the injection is 10-20 mM; the concentration of the sucrose in the injection is 85-95 mg / mL; the concentration of the polysorbate 80 in the injection is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1-0.5 mg / mL; the pH value of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0080] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0081] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: 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 is 25-50 mg / mL; the concentration of the histidine buffer in the injection is 10-20 mM; the concentration of the trehalose in the injection is 55-65 mg / mL; the concentration of the polysorbate 80 in the injection is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1-0.5 mg / mL; the pH value of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0082] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0083] In another aspect of the present invention, the present invention provides an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, sucrose or trehalose, and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 mg / mL; The concentration of the citrate buffer in the injection is 8-25mM, preferably 10-20mM; the concentration of the sucrose in the injection is 85-95mg / mL; the concentration of the trehalose in the injection is 55-65mg / mL; the concentration of the polysorbate 80 in the injection is 0.4-1.3mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1-0.5mg / mL; the pH value of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible foreign matter, improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0084] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0085] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 mg / mL; the citrate buffer is The concentration of the injection is 8-25mM, preferably 10-20mM; the concentration of mannitol in the injection is 35-45mg / mL; the concentration of polysorbate 80 in the injection is 0.4-1.5mg / mL, or 0.4-1.3mg / mL, or the concentration of polysorbate 20 in the injection is 0.1-0.5mg / mL; the concentration of the metal chelator in the injection is 0.05-0.5mg / mL; the pH value of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible foreign matter, improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0086] According to an embodiment of the present invention, the injection does not contain disodium edetate or calcium sodium edetate. In another aspect of the present invention, the present invention provides an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80 and optionally disodium edetate or calcium sodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection is 5 to 20 mM; the concentration of mannitol in the injection is 40 to 50 mg / mL; the concentration of polysorbate 80 in the injection is 0.1 to 1.5 mg / mL; the concentration of the metal chelator in the injection is 0.05 to 0.5 mg / mL; the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0087] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0088] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80 and optionally disodium edetate or calcium sodium edetate; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection is 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of mannitol in the injection is 40 to 50 mg / mL; the concentration of polysorbate 80 in the injection is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the injection is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0089] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0090] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80 and optionally disodium edetate or calcium sodium 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection is 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of mannitol in the injection is 40 to 50 mg / mL; the concentration of polysorbate 80 in the injection is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the injection is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0091] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0092] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80 and optionally disodium edetate or sodium calcium edetate, and sucrose or trehalose; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection is 5 to 20 mM; the concentration of the sucrose in the injection is 80 to 95 mg / mL; the concentration of the trehalose in the injection is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection is 0.1 to 1.5 mg / mL; and the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0093] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0094] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer and polysorbate 80, as well as sucrose or trehalose, and optionally disodium edetate or calcium sodium 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection is 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of the sucrose in the injection is 80 to 95 mg / mL; the concentration of the trehalose in the injection is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection is 0.1 to 0.5 mg / mL; and the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0095] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0096] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0097] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0098] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0 to 7 mg / mL, preferably 3 to 7 mg / mL.

[0099] In an optional embodiment of the present application, the concentration of m-cresol in the composition or injection is 0-4 mg / mL, preferably 2-4 mg / mL.

[0100] In another aspect of the present invention, the present invention provides a use of the aforementioned composition or the aforementioned injection in the preparation of a medicament for treating and / or preventing metabolic diseases.

[0101] According to an embodiment of the present invention, the metabolic diseases include diabetes, hyperlipidemia, obesity and diseases related to fatty liver.

[0102] According to an embodiment of the present invention, the fatty liver-related diseases include non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and liver cirrhosis.

[0103] In another aspect of the present invention, the present invention provides an injection device or container. According to an embodiment of the present invention, the injection device or container comprises: the aforementioned composition or the aforementioned injection solution.

[0104] According to an embodiment of the present invention, the injection device or container includes at least one selected from a container bottle and a syringe.

[0105] According to an embodiment of the present invention, the syringe includes at least one selected from an automatic syringe and a pre-filled syringe.

[0106] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0107] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0108] Figure 1 This is a high temperature SEC-aggregate growth trend diagram of different preparations in step 7 of Example 1 of the present invention;

[0109] Figure 2 This is a graph showing the growth trend of the high temperature RP-HPLC-purity (total related substances) of different preparations in step 7 of Example 1 of the present invention. DETAILED DESCRIPTION

[0110] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0111] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

[0112] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0113] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

[0114] As used herein, the terms "optionally", "optional" or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0115] As used herein, the term "treatment" refers to the use of drugs to obtain the desired pharmacological and / or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, especially humans, including: (a) preventing the occurrence of a disease or condition in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as blocking the progression of the disease; or (c) alleviating the disease, such as alleviating symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a composition, injection or drug to an individual to treat, cure, alleviate, improve, reduce or inhibit an individual's disease, including but not limited to administering a drug containing a composition or injection described herein to an individual in need.

[0116] The present invention provides a composition and an injection solution, and uses thereof and an injection device or container, which will be described in detail below.

[0117] Composition

[0118] In one aspect of the present invention, the present invention provides a composition. According to an embodiment of the present invention, the composition comprises: a GLP-1-Fc-FGF21 fusion protein and a buffer. In the composition of the present invention, by adding a buffer, the generation of visible foreign matter and aggregates can be effectively controlled, the stability of the GLP-1-Fc-FGF21 fusion protein can be improved, and thus the shelf life of the product can be extended.

[0119] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NOs: 1-12.

[0120] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(SEQ ID NO:1);

[0121] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(SEQ ID NO:2);

[0122] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPAVFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(SEQ ID NO:3);

[0123] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYES(SEQ ID NO:4);

[0124] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTECHLEIREDGTVGCAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(SEQ ID NO:5);

[0125] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTECHLEIREDGTVGCAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(SEQ ID NO:6);

[0126] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYES(SEQ ID NO:7);

[0127] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(SEQ ID NO:8);

[0128] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQRYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(SEQ ID NO:9);

[0129] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQQYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPAQFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGGSQGLSPSYAS(SEQ ID NO:10);

[0130] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQQYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQILGVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPAQFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYAS(SEQ ID NO:11);

[0131] HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTK PREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSHPIPDSSPLLQFGGQVRQVYLYTDDAQQTEAHLEIREDGTVGGAADQSPESLLQLKALKPGVIQIL GVKTSRFLCQRPDGALYGSLHFDPEACSFRERLLEDGYNVYQSEAHGLPLHLPGNKSPHRDPAPRGPARFLPLPGLPPALPEPPGILAPQPPDVGSSDPLHMVGASQGLSPSYES(SEQ ID NO:12).

[0132] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in any one of SEQ ID NOs: 1 to 12.

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

[0134] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:2.

[0135] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:3.

[0136] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:4.

[0137] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:5.

[0138] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:6.

[0139] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:7.

[0140] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:8.

[0141] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:9.

[0142] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:10.

[0143] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:11.

[0144] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO:12.

[0145] In some optional embodiments of the present invention, the concentration of the 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, 85 mg / mL, 90 mg / mL, 100 mg / mL or a range value between any two values ​​as endpoint values.Such as 2 to 99 mg / mL, 2 to 95 mg / mL, 2 to 90 mg / mL, 2 to 85 mg / mL, 2 to 80 mg / mL, 2 to 75 mg / mL, 2 to 70 mg / mL, 2 to 65 mg / mL, 2 to 60 mg / mL, 2 to 55 mg / mL, 5 to 99 mg / mL, 5 to 95 mg / mL, 5 to 90 mg / mL, 5 to 85 mg / mL, 5 to 80 mg / mL, 5 to 75 mg / mL, 5 to 70 mg / mL, 5 to 65 mg / mL, 5 to 60 mg / mL, 5 to 55 mg / mL, 10 to 99 mg / mL, 10 to 95 mg / mL, 10 to 90 mg / mL, 10 to 85 mg / mL, 10 to 80 mg / mL, 10 to 75 mg / mL, 10 to 70 mg / mL, 10 to 65 mg / mL, 10 to 60 mg / mL, 10 to 55 mg / mL, 25 to 99 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, 30 to 99 mg / mL, 30 to 95 mg / mL, 30 to 90 mg / mL, 30 to 85 mg / mL, 30 to 80 mg / mL, 30 to 75 mg / mL, 30 to 70 mg / mL, 30 to 65 mg / mL, 30 to 60 mg / mL, 30 to 55 mg / mL, 35 to 99 mg / mL, 35 to 95 mg / mL, 35 to 90 mg / mL, 35 to 85 mg / mL, 35 to 80 mg / mL, 35 to 75 mg / mL, 35 to 70 mg / mL, 35 to 65 mg / mL, 35 to 60 mg / mL, 35 to 55 mg / mL, 40 to 99 mg / mL, 40 to 95 mg / mL, 40 to 90 mg / mL, 40 to 85 mg / mL, 40 to 80 mg / mL, 40 to 75 mg / mL, 40 to 70 mg / mL, 40 to 65 mg / mL, 40 to 60 mg / mL, 40 to 55 mg / mL.,

[0146] In some optional embodiments of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the composition is 2-55 mg / mL, for example, 2-25 mg / mL, 3-25 mg / mL, 10-25 mg / mL, 10-20 mg / mL, 15-25 mg / mL, 15-20 mg / mL, 25-100 mg / mL, 25-95 mg / mL, 25-90 mg / mL, 25-85 mg / mL, 25-80 mg / mL, 25-75 mg / mL, 25-70 mg / mL, 25-65 mg / mL, 25-60 mg / mL, 25-55 mg / mL, 25-50 mg / mL, 25-40 mg / mL, 40-55 mg / mL, 40-50 mg / mL, 45-55 mg / mL, 45-50 mg / mL.

[0147] In some optional embodiments of the present invention, the pH value of the composition is 6.4-8.0, such as 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 a range value between any two values ​​as endpoint values. Thus, the stability of the fusion protein can be further improved, such as 6.2-8.0, 6.9-7.5.

[0148] In some optional embodiments of the present invention, the buffer comprises at least one of a histidine buffer, a citrate buffer, a phosphate buffer and a Tris buffer.

[0149] In this document, without special explanation, "citrate buffer" or "citrate buffer" can be selected from citric acid, sodium citrate, or a mixture of citric acid and sodium citrate, all of which are within the protection scope of the present application.

[0150] 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.

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

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

[0153] In some optional embodiments of the present invention, the histidine buffer comprises histidine or histidine hydrochloride.

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

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

[0156] In some optional embodiments of the present invention, the citrate buffer comprises citric acid or sodium citrate.

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

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

[0159] In some optional embodiments of the present invention, the Tris buffer comprises Tris or Tris-HCl.

[0160] In some optional embodiments of the present invention, the buffer comprises histidine buffer or citrate buffer.

[0161] In some optional embodiments of the present invention, the concentration of the buffer in the composition is 5-30mM, for example 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 11mM, 11.29mM, 12mM, 13mM, 14mM, 15mM, 16mM, 17mM, 18mM, 19mM, 20mM, 21mM, 22mM, 23mM, 24mM, 25mM, 26mM, 27mM, 28mM, 29mM or 30mM, or a range value between any two values ​​as endpoint values ​​(for example 5-25mM, 8-25mM, 8-12mM, 10-20mM).

[0162] 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 chelator, thereby further improving the stability of the fusion protein.

[0163] In some optional embodiments of the present invention, the osmotic pressure regulator includes at least one of mannitol, sucrose, sodium chloride and trehalose.

[0164] In some optional embodiments of the present invention, the concentration of the osmotic pressure regulator in the composition is 30-100 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 100 mg / mL, or a range between any two values ​​as endpoints.

[0165] In some optional embodiments of the present invention, the concentration of the osmotic pressure 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 a range value between any two values ​​as endpoint values.

[0166] In some optional embodiments of the present invention, the surfactant includes at least one of polysorbate 80, polysorbate 20 and poloxamer 188.

[0167] 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 In some embodiments, the present invention relates to a method for preparing the present invention which is preferably used as a method for preparing the present invention. The method may be used in any one of the following embodiments:

[0168] Illustratively, the concentration of the surfactant in the composition is 0.1 to 1.5 mg / mL or 0.1 to 1 mg / mL.

[0169] In some optional embodiments of the present invention, the preservative is at least one of phenol and m-cresol.

[0170] According to an embodiment of the present invention, the concentration of the preservative in the composition is 0.1-10 mg / mL, for example 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 1.5 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, 10.0 mg / mL, such as 1.0-5.0 mg / mL.

[0171] In some optional embodiments of the present invention, the preservative is phenol.

[0172] According to an embodiment 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, 10.0 mg / mL, such as 2 to 7 mg / mL, 3.0 to 4.0 mg / mL.

[0173] In some optional embodiments of the present invention, the preservative is m-cresol.

[0174] According to an embodiment of the present invention, the concentration of meta-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, 10.0 mg / mL, such as 3.0 to 4.0 mg / mL.

[0175] In some optional embodiments of the present invention, the preservatives are phenol and m-cresol.

[0176] According to an embodiment of the present invention, the concentration of phenol in the composition is 1.0 to 5.0 mg / mL (e.g., 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, 10.0 mg / mL), and the concentration of m-cresol in the composition is 0.1-1.0 mg / mL (e.g., 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).

[0177] In some optional embodiments of the present invention, the metal chelator is at least one of edetate disodium (EDTA-2Na) and edetate sodium calcium (EDTA-CaNa2).

[0178] In some optional embodiments of the present invention, the metal chelator is edetate disodium (EDTA-2Na).

[0179] In some optional embodiments of the present invention, the metal chelator is edetate sodium calcium (EDTA-CaNa2).

[0180] In some optional embodiments of the present invention, the concentration of the metal chelator in the composition is 0.01-1 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, 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, or a range value between any two values ​​as the endpoint values ​​(0.05-0.5 mg / mL, 0.08-0.12 mg / mL, 0.1-0.12 mg / mL).

[0181] Exemplarily, the concentration of the metal chelator in the composition is 0.08-0.12 mg / mL. In the embodiments of the present invention, whether it is a citric acid system or a histidine buffer system, whether EDTA sodium salt is added to the preparation of the fusion protein, the stability of the preparation is good. That is, whether EDTA sodium salt is added is within the scope of protection of the present invention.

[0182] In some optional embodiments 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, 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, 100 mg / mL, or a range value between any two values ​​as endpoint values ​​(e.g., 25 to 75 mg / mL, 25~70mg / mL, 25~65mg / mL, 25~60mg / mL, 25~55mg / mL, 25~50mg / mL, 25~40mg / mL, 40~80mg / mL, 40~75mg / mL, 40~70mg / mL, 40~65mg / mL, 40~60mg / mL, 40~55mg / mL, 40~50mg / mL, 45~80mg / mL, 45~75mg / mL, 45~70mg / mL, 45~65mg / mL, 45~60mg / mL, 45~55mg / mL, 45~50mg / mL).

[0183] 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 a histidine buffer. The inventor unexpectedly found that the final concentration of the fusion protein is 25-55 mg / mL. By adding the histidine buffer, the production of visible foreign matter in the composition can be effectively controlled, which can not only improve the physical stability of the fusion protein and reduce potential safety risks, but also effectively control the rate of chemical changes, maintain stability under high temperature, accelerated, long-term, repeated freeze-thaw, shock and simulated transportation conditions, and the prepared composition has a longer shelf life.

[0184] In some optional embodiments of the present invention, the concentration of the histidine buffer in the composition is 5-30mM, for example 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 11mM, 12mM, 13mM, 14mM, 15mM, 16mM, 17mM, 18mM, 19mM, 20mM, 21mM, 22mM, 23mM, 24mM, 25mM, 26mM, 27mM, 28mM, 29mM, 30mM, or a range value between any two values ​​as endpoint values ​​(e.g., 5-25mg / mL, 8-25mg / mL, 8-20mg / mL, 10-20mg / mL, 8-12mg / mL).

[0185] In some optional embodiments of the present invention, the concentration of the histidine buffer in the composition is 10-20 mM.

[0186] In some optional embodiments of the present invention, the pH value of the composition is 6.7-7.5, preferably 6.9-7.5.

[0187] In some optional embodiments of the present invention, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30-50 mg / mL; or the osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60-100 mg / mL, preferably 80-100 mg / mL; or the osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40-100 mg / mL, preferably 40-70 mg / mL, 50-70 mg / mL, 55-65 mg / mL; or the osmotic pressure regulator is sodium chloride, and the concentration of the osmotic pressure regulator in the composition is 1-10 mg / mL, preferably 3-9 mg / mL.

[0188] In some optional embodiments of the present invention, the surfactant is polysorbate 80, and the concentration of the surfactant in the composition is 0.1-2 mg / mL, such as 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 a range value between any two values ​​as endpoint values ​​(e.g. 0.1-1.5 mg / mL, 0.2-2 mg / mL, 0.4-1.3 mg / mL); or

[0189] In some optional embodiments of the present invention, the surfactant is polysorbate 20, and the concentration of the surfactant in the composition is 0.1-1.5 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 a range value between any two values ​​as endpoint values ​​(for example, 0.1-0.5 mg / mL).

[0190] 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 inventor unexpectedly discovered that the final concentration of the fusion protein is 2 to 25 mg / mL, and the addition of a citrate buffer can effectively control the generation of visible foreign matter in the composition, improve the physical and chemical stability of the fusion protein, reduce potential safety risks, and extend the shelf life of the composition.

[0191] In some optional embodiments of the present invention, the concentration of the citrate buffer in the composition is 5-30mM, for example, 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 11mM, 12mM, 13mM, 14mM, 15mM, 16mM, 17mM, 18mM, 19mM, 20mM, 21mM, 22mM, 23mM, 24mM, 25mM, 26mM, 27mM, 28mM, 29mM or 30mM, or a range value between any two values ​​as endpoint values ​​(for example, 5-25mM, 8-25mM, 8-20mM, 10-20mM, 8-12mM, 5-15mM, 10-15mM).

[0192] In some optional embodiments of the present invention, the pH value of the composition is 6.4-8.0.

[0193] In some optional embodiments of the present invention, the osmotic pressure regulator is mannitol, and the concentration of the osmotic pressure regulator in the composition is 30-50 mg / mL; or the osmotic pressure regulator is sucrose, and the concentration of the osmotic pressure regulator in the composition is 60-100 mg / mL, preferably 80-100 mg / mL; or the osmotic pressure regulator is trehalose, and the concentration of the osmotic pressure regulator in the composition is 40-100 mg / mL, preferably 40-70 mg / mL, and more preferably 50-70 mg / mL.

[0194] According to an 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.

[0195] In some optional embodiments of the present invention, the surfactant includes at least one of polysorbate 80, polysorbate 20 and poloxamer 188, preferably polysorbate 80.

[0196] 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 a range value between any two values ​​as endpoint values ​​(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, 0.1 to 0.3 mg / mL).

[0197] In some optional embodiments 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 mg / mL or 0.1 to 0.5 mg / mL.

[0198] In some optional embodiments of the present invention, the composition is in the form of an injection.

[0199] Injection

[0200] In another aspect of the present invention, the present invention provides an injection solution. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as 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 is 25 to 100 mg / mL; the concentration of the histidine buffer in the composition is 8 to 25 mM; the concentration of mannitol in the composition is 35 to 45 mg / mL; the concentration of polysorbate 80 in the composition is 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the composition is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the composition is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter and aggregates, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0201] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 95 mg / mL.

[0202] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25 to 90 mg / mL.

[0203] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25-85 mg / mL.

[0204] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 80 mg / mL.

[0205] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 75 mg / mL.

[0206] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-70 mg / mL.

[0207] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 65 mg / mL.

[0208] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-60 mg / mL.

[0209] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-55 mg / mL.

[0210] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL.

[0211] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 12.

[0212] According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 1, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25 to 50 mg / mL; the concentration of the histidine buffer in the composition is 10 to 20 mM; the concentration of mannitol in the composition is 35 to 45 mg / mL; the concentration of polysorbate 80 in the composition is 0.4 to 1.3 mg / mL, or the concentration of polysorbate 20 in the composition is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the composition is 0.05 to 0.5 mg / mL; the pH value of the injection is 6.9 to 7.5.

[0213] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0214] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0215] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0216] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0-7 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, or a range value between any two values ​​as endpoint values ​​(for example, 3-7 mg / mL, 2-4 mg / mL).

[0217] In an optional embodiment of the present application, the concentration of meta-cresol in the composition or injection is 0-4 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 a range value between any two values ​​as endpoint values ​​(for example, 2-4 mg / mL).

[0218] In another aspect of the present invention, the present invention provides an injection solution. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer, sucrose or trehalose, and optionally disodium edetate or calcium sodium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as 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 is 25 to 100 mg / mL; the concentration of the histidine buffer in the composition is 8 to 20 mM; the concentration of the sucrose in the composition is 85 to 95 mg / mL; the concentration of the trehalose in the injection is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the composition is 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the composition is 0.1 to 0.5 mg / mL; the pH value of the injection is 6.9 to 7.5. The injection of the present invention can effectively control the generation of visible foreign matter and aggregates, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0219] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 95 mg / mL.

[0220] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25 to 90 mg / mL.

[0221] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25-85 mg / mL.

[0222] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 80 mg / mL.

[0223] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 75 mg / mL.

[0224] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-70 mg / mL.

[0225] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25 to 65 mg / mL.

[0226] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-60 mg / mL.

[0227] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-55 mg / mL.

[0228] According to an embodiment of the present invention, the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection solution is 25-50 mg / mL.

[0229] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 12.

[0230] According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, histidine buffer and 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, and the concentration of the GLP-1-Fc-FGF21 fusion protein in the injection is 25-50 mg / mL; the concentration of the histidine buffer in the injection is 10-20 mM; the concentration of the sucrose in the injection is 85-95 mg / mL; the concentration of the polysorbate 80 in the injection is 0.4-1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1-0.5 mg / mL; the pH value of the injection is 6.9-7.5.

[0231] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0232] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0233] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0234] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0-7 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, or a range value between any two values ​​as endpoint values ​​(for example, 3-7 mg / mL, 2-4 mg / mL).

[0235] In an optional embodiment of the present application, the concentration of meta-cresol in the composition or injection is 0-4 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 a range value between any two values ​​as endpoint values ​​(for example, 2-4 mg / mL).

[0236] In another aspect of the present invention, the present invention provides an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, sucrose or trehalose, and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 mg / mL; The concentration of the citrate buffer in the injection is 8-25mM, preferably 10-20mM; the concentration of the sucrose in the injection is 85-95mg / mL; the concentration of the trehalose in the injection is 55-65mg / mL; the concentration of the polysorbate 80 in the injection is 0.4-1.3mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1-0.5mg / mL; the pH value of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible foreign matter, improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0237] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0238] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0239] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0240] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0-7 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, or a range value between any two values ​​as endpoint values ​​(for example, 3-7 mg / mL, 2-4 mg / mL).

[0241] In an optional embodiment of the present application, the concentration of meta-cresol in the composition or injection is 0-4 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 a range value between any two values ​​as endpoint values ​​(for example, 2-4 mg / mL).

[0242] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol and optionally disodium edetate or sodium calcium edetate, and polysorbate 80 or polysorbate 20; the GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 mg / mL; the citrate buffer is The concentration of the injection is 8-25mM, preferably 10-20mM; the concentration of mannitol in the injection is 35-45mg / mL; the concentration of polysorbate 80 in the injection is 0.4-1.5mg / mL, or 0.4-1.3mg / mL, or the concentration of polysorbate 20 in the injection is 0.1-0.5mg / mL; the concentration of the metal chelator in the injection is 0.05-0.5mg / mL; the pH value of the injection is 6.9-7.5. The injection of the present invention can effectively control the generation of visible foreign matter, improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0243] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0244] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0245] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0246] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0-7 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, or a range value between any two values ​​as endpoint values ​​(for example, 3-7 mg / mL, 2-4 mg / mL).

[0247] In an optional embodiment of the present application, the concentration of meta-cresol in the composition or injection is 0-4 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 a range value between any two values ​​as endpoint values ​​(for example, 2-4 mg / mL).

[0248] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80 and optionally disodium edetate or calcium sodium edetate; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the composition is 5 to 20 mM; the concentration of mannitol in the composition is 40 to 50 mg / mL; the concentration of polysorbate 80 in the composition is 0.1 to 1.5 mg / mL; the concentration of the metal chelator in the composition is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter and aggregates, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0249] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 12.

[0250] In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection is 8 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection is 10 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection is 10 to 15 mM.

[0251] According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80 and optionally disodium edetate or calcium sodium 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection is 5 to 25 mM, 8 to 20 mM, 10 to 20 mM, or 10 to 15 mM; the concentration of mannitol in the injection is 40 to 50 mg / mL; the concentration of polysorbate 80 in the injection is 0.1 to 0.5 mg / mL; the concentration of the metal chelator in the injection is 0.05 to 0.5 mg / mL; and the pH value of the injection is 6.4 to 8.0.

[0252] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0253] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0254] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0255] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0-7 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, or a range value between any two values ​​as endpoint values ​​(for example, 3-7 mg / mL, 2-4 mg / mL).

[0256] In an optional embodiment of the present application, the concentration of meta-cresol in the composition or injection is 0-4 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 a range value between any two values ​​as endpoint values ​​(for example, 2-4 mg / mL).

[0257] In another aspect of the present invention, the present invention proposes an injection. According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer, polysorbate 80 and optionally disodium edetate or sodium calcium edetate, and sucrose or trehalose; the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as 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 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; the pH value of the injection is 6.4 to 8.0. The injection of the present invention can effectively control the generation of visible foreign matter and aggregates, and can improve the physical stability and chemical stability of the fusion protein, thereby reducing potential safety risks and extending the shelf life.

[0258] In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 1. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 2. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 3. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 4. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 5. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 6. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 7. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 8. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 9. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 10. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 11. In some optional embodiments of the present invention, the GLP-1-Fc-FGF21 fusion protein has an amino acid sequence as shown in SEQ ID NO: 12.

[0259] In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection is 8 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection is 10 to 20 mM. In some optional embodiments of the present invention, the concentration of the citrate buffer in the injection is 10 to 15 mM.

[0260] According to an embodiment of the present invention, the injection comprises: GLP-1-Fc-FGF21 fusion protein, citrate buffer and polysorbate 80, as well as sucrose or trehalose, and optionally disodium edetate or sodium calcium 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 is 3 to 25 mg / mL; the concentration of the citrate buffer in the injection 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 is 80 to 95 mg / mL; the concentration of the trehalose in the injection is 55 to 65 mg / mL; the concentration of the polysorbate 80 in the injection is 0.1 to 0.5 mg / mL; and the pH value of the injection is 6.4 to 8.0.

[0261] According to an embodiment of the present invention, the injection does not contain edetate disodium or edetate calcium sodium.

[0262] In an optional embodiment of the present application, the injection solutions of the above aspects may further include a preservative.

[0263] In an optional embodiment of the present application, the preservative is at least selected from phenol, m-cresol, benzyl alcohol, and phenoxyethanol.

[0264] In an optional embodiment of the present application, the concentration of phenol in the composition or injection is 0-7 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, or a range value between any two values ​​as endpoint values ​​(for example, 3-7 mg / mL, 2-4 mg / mL).

[0265] In an optional embodiment of the present application, the concentration of meta-cresol in the composition or injection is 0-4 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 a range value between any two values ​​as endpoint values ​​(for example, 2-4 mg / mL).

[0266] use

[0267] In another aspect of the present invention, the present invention provides a use of the aforementioned composition or the aforementioned injection in the preparation of a medicament for treating and / or preventing metabolic diseases.

[0268] In some optional embodiments of the present invention, the metabolic diseases include diabetes, obesity, hyperlipidemia and diseases related to fatty liver.

[0269] In some optional embodiments of the present invention, the fatty liver-related diseases include non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and cirrhosis.

[0270] Methods for treating and / or preventing metabolic diseases

[0271] In another aspect of the present invention, the present invention provides a method for treating and / or preventing metabolic diseases. According to an embodiment of the present invention, the method comprises: administering a pharmaceutically acceptable amount of the aforementioned composition or the aforementioned injection to a subject.

[0272] The effective amount of the composition or injection of the present invention may vary with the mode of administration and the severity of the disease to be treated. The selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to: pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated, the patient's body weight, the patient's immune status, the route of administration, etc. For example, according to the urgent requirements of the treatment condition, the dosage may be administered once a day, once every other day, once a week, or once every two weeks.

[0273] In some optional embodiments of the present invention, the administration route of the method includes subcutaneous injection or intravenous injection, preferably subcutaneous injection.

[0274] Injection device or container

[0275] In another aspect of the present invention, the present invention provides an injection device or container. According to an embodiment of the present invention, the injection device or container comprises: the aforementioned composition or the aforementioned injection solution.

[0276] In some optional embodiments of the present invention, the injection device or container includes at least one selected from a container bottle, a vial, a cartridge and a syringe.

[0277] In some optional embodiments of the present invention, the syringe comprises at least one selected from an automatic syringe and a pre-filled syringe.

[0278] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0279] It should be noted that the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein in Example 1 of the present invention is shown in SEQ ID NO: 1, and is prepared by conventional methods in the art.

[0280] Embodiment 1:

[0281] 1. Investigation of injection solutions prepared with different buffer systems

[0282] 1.1 Different buffer systems were used to prepare low-concentration injection preparations A1 to A3. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 1. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the prescription amount in Table 1. If necessary, the pH of the preparation is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally water for injection is added to make up the volume. After the liquid is prepared, the liquid is filtered in series with 0.45μm and 0.22μm PVDF filter membranes into a 1ml slender prefilled syringe, each filled with 1ml, and sealed with a rubber stopper. Then the preparations A1 to A3 were subjected to high temperature stability studies. The SEC-HPLC stability test results of 0 days, 10 days and 14 days under high temperature conditions of 40°C are shown in Table 2.

[0283] Table 1: Ratios of ingredients in formulations A1 to A3

[0284]

[0285] Table 2: Stability results of formulations A1 to A3 at high temperature 40°C

[0286]

[0287]

[0288] Note: (1) The visible foreign matter in the above table and in the following text that is in compliance with the regulations means that it complies with the provisions of General Rule 0904 of Part IV of ChP 2015. The visible foreign matter that is not in compliance with the regulations means that it does not comply with the provisions of General Rule 0904 of Part IV of ChP 2015.

[0289] (2) The reason why the main peak purity of RP-HPLC-purity (main peak) at day 0 in the above table is relatively low is mainly because the reverse phase main peak purity of the API stock solution directly used in this example is not high. Therefore, the data in this example is only used for screening of preparation prescriptions and does not represent the purity of the actual fusion protein sample of this application, the same as preparations A4 to A37 in this example.

[0290] As shown in Table 2, the RP-HPLC main peak purity decrease rate in the citrate buffer system is the slowest, and at a high temperature of 40°C and 14 days, the visible foreign matter meets the requirements. Therefore, for low-concentration fusion protein preparations, the chemical stability and physical stability of the citrate buffer system are relatively good.

[0291] Since different buffer systems may also have different effects on the thermal stability of proteins, the Tm values ​​and Tonset values ​​of preparations A1 to A3 in different buffer systems were further determined by DSC (differential scanning calorimetry) to compare the differences in the effects of various factors on the thermal stability of proteins by studying the temperature changes during the thermal denaturation process of proteins. The DSC test results are shown in Table 3. Among them, Tonset and Tm values ​​are indicators of thermal stability, Tonset value is the starting temperature of protein unfolding, and Tm value is the midpoint temperature of protein thermal transition. Generally speaking, the higher the Tonset value and Tm value, the more stable the protein is, and the less likely it is to unfold and denature in a low temperature environment.

[0292] Table 3: DSC test results of preparations A1 to A3

[0293] preparation pH <![CDATA[T onset (℃)]]> <![CDATA[T m1 (℃)]]> <![CDATA[T m2 (℃)]]> A1 6.5 60.92 66.78 84.40 A2 6.5 60.23 66.64 83.71 A3 6.5 56.36 64.39 83.91

[0294] As shown in Table 3, by comparing the Tonset, Tm1 and Tm2 values, for low concentration fusion protein preparations, the thermal stability in citrate (preparation A1) and phosphate buffer (preparation A2) is better than that in the histidine buffer system (preparation A3).

[0295] 1.2 Different buffer systems were used to prepare high-concentration injection preparations F1 to F4. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein) in the injection and the types and final concentrations of each excipient are specifically shown in Table 4. The preparation method of the injection is: after ultrafiltration and concentration of the fusion protein stock solution, the protein concentration is detected, and then the fusion protein solution is diluted to 25 mg / ml with the corresponding buffer system. If necessary, 2% hydrochloric acid or 2% sodium hydroxide solution is used to adjust the pH to 7.0. After the liquid is prepared, the liquid is filtered in series with 0.45μm and 0.22μm PVDF filter membranes into 2R medium borosilicate vials, each with a volume of 1ml, plugged with a rubber stopper, and sealed with an aluminum-plastic combination cap. Then, the preparations F1 to F4 were subjected to high temperature stability studies. The results of the SEC-HPLC aggregation change rate (days / %) at high temperature of 40°C for 14 days are shown in Tables 5-1, 5-2 and 5-3.

[0296] Table 4: Ratios of ingredients in formulations F1 to F4

[0297]

[0298] Table 5-1: SEC-HPLC aggregation change rate of preparations F1 to F4 placed at high temperature 40°C for 14 days (days / %)

[0299]

[0300]

[0301] Table 5-2: IEX-HPLC main peak change rate of preparations F1 to F4 placed at 40°C for 14 days (days / %)

[0302] preparation 0 days - IEX main peak (%) High temperature 40℃ 14 days-IEX main peak (%) Change rate (% / day) F1 58.69 41.02 1.26 F2 56.90 42.56 1.02 F3 57.04 35.91 1.51 F4 55.73 43.10 0.90

[0303] Table 5-3: RP-HPLC total impurity change rate of preparations F1 to F4 placed at 40°C for 14 days (days / %)

[0304] preparation Day 0-RP-HPLC total impurities (%) High temperature 40℃14 days-RP-HPLC Total impurities (%) Change rate (% / day) F1 21.80 31.06 0.66 F2 22.30 33.99 0.83 F3 20.54 31.34 0.77 F4 22.18 30.64 0.60

[0305] As shown in Table 5-1, after 14 days at high temperature of 40°C, the SEC aggregates grew faster in the sodium citrate buffer (preparation F1), phosphate buffer (preparation F3) and Tris buffer (preparation F4) systems, and the histidine buffer (preparation F2) was able to significantly reduce the growth rate of the SEC aggregates (SEC aggregate standard is 5%); the RP-HPLC total impurities (total impurities standard 40%), the four preparations had similar change rates. Therefore, it can be further explained that when the concentration of the fusion protein is greater than or equal to 25 mg / ml, the stability of the preparation prepared with the histidine buffer is higher, but the stability data of the preparations in different buffer systems are all within the acceptable standard range.

[0306] 2. Investigation of injection solutions prepared at different pH

[0307] 2.1 Low-concentration injection preparations A4 to A8 were prepared using different pH values. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 6. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the prescription amount in the following table. If necessary, the pH of the preparation is adjusted to the corresponding value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution, and finally the injection water is added to the volume. After the preparation is completed, the drug solution is filtered in series with 0.45μm and 0.22μm PVDF filter membranes into a 1ml slender prefilled syringe, each filled with 1ml, and sealed with a rubber stopper. Then, the preparations A4 to A8 were subjected to high temperature stability and long-term stability studies. The SEC-HPLC stability test results of 0 days, 10 days, and 14 days at high temperature of 40°C are shown in Table 7, and the SEC-HPLC stability test results of 0 months, 1 month, 6 months, and 12 months at 4°C refrigerator conditions are shown in Table 8.

[0308] Table 6: Ratios of ingredients in formulations A4 to A8

[0309]

[0310] Table 7: Stability results of formulations A4 to A8 at high temperature 40°C

[0311]

[0312]

[0313] Table 8: Stability results of formulations A4 to A8 at 4°C

[0314]

[0315]

[0316] It can be seen from Tables 7 and 8 that when the pH is 6.0, the RP-HPLC-main peak purity of the injection under high temperature 40°C conditions has the fastest decrease rate, the dimer content is the highest, and the stability is the worst. In addition, the long-term visible foreign matter for 12 months does not meet the regulations, the SEC-purity (monomer including shoulder peak) decreases at the fastest rate, the dimer content is the highest, and the stability is the worst. The chemical stability of the other pH value groups is good.

[0317] Since different buffer systems may also have different effects on the thermal stability of proteins, the Tm values ​​and Tonset values ​​of preparations A4 to A8 in different buffer systems were further determined by DSC (differential scanning calorimetry) to compare the effects of various factors on the thermal stability of proteins by studying the temperature changes during the thermal denaturation of proteins. The DSC test results are shown in Table 9.

[0318] Table 9: DSC test results of preparations A4 to A8

[0319] preparation pH Tonset(℃) Tm1(℃) Tm2(℃) A4 6.0 57.86 64.79 84.29 A5 6.5 60.62 66.49 84.37 A6 7.0 60.78 67.45 84.00 A7 7.5 59.92 67.94 83.69 A8 8.0 60.35 67.84 83.58

[0320] From the DSC test results in Table 9, it can be seen that the thermal stability is the worst when the pH is 6.0, and the other groups are all good.

[0321] The effect of pH between 6.0 and 7.0 on product stability was further refined. Injection preparations A9 to A15 were prepared using different pH values. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically shown in Table 10. The preparation method of the injection is specifically referred to the preparation method of preparations A4 to A8 in step 2.1 of Example 1. Then, preparations A9 to A15 were subjected to high temperature stability study and DSC testing. The SEC-HPLC stability test results at high temperature of 40°C for 0 days, 10 days, 14 days and 21 days are shown in Table 11, and the DSC test results are shown in Table 12.

[0322] Table 10: Ratios of ingredients in formulations A9 to A15

[0323]

[0324] Table 11: Stability results of formulations A9-A15 at high temperature 40°C

[0325]

[0326]

[0327] Table 12: DSC test results of preparations A9 to A15

[0328] preparation pH <![CDATA[T onset (℃)]]> <![CDATA[T m1 (℃)]]> <![CDATA[T m2 (℃)]]> A9 6.0 58.96 64.57 84.31 A10 6.2 59.04 65.45 84.39 A11 6.4 60.36 66.50 84.37 A12 6.5 60.24 66.65 84.26 A13 6.6 60.81 66.94 84.29 A14 6.8 59.93 67.40 83.94 A15 7.0 59.80 67.55 84.09

[0329] It can be seen from Tables 11 and 12 that when the pH is 6.0 and 6.2, the Tonset and Tm1 values ​​are low, and the thermal stability is worse than that of the other groups. When the pH is 6.4 and above, the chemical stability, physical stability and thermal stability of the injection are good.

[0330] 2.2 High-concentration injection preparations F5 to F11 were prepared using different pH values. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 13. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the prescription amount in the following table. If necessary, the pH of the preparation is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally the volume is fixed to 10 ml with injection water. After the preparation is completed, the drug solution is filtered in series with 0.45 μm and 0.22 μm PVDF filter membranes into a 1 ml slender prefilled syringe, with a volume of 0.5 ml per syringe, and a rubber stopper is plugged and sealed. Then the preparations F1 to F4 were subjected to a high temperature stability study. The results of the SEC-HPLC aggregation change rate (days / %) at a high temperature of 40°C for 20 days are shown in Table 14.

[0331] Table 13: Ratios of ingredients in formulations F5 to F11

[0332]

[0333]

[0334] Table 14: SEC-HPLC aggregation change rate of preparations F5 to F11 placed at high temperature 40°C for 20 days (days / %)

[0335] preparation Day 0-SEC Aggregates (%) High temperature 40℃20 days-SEC aggregates (%) Change rate (% / day) F5 1.46 5.53 0.20 F6 1.44 4.96 0.18 F7 1.01 3.74 0.14 F8 0.89 2.76 0.09 F9 0.93 2.55 0.08 F10 0.88 2.21 0.07 F11 0.86 2.06 0.06

[0336] As shown in Table 14, after being placed at high temperature of 40°C for 20 days, within the pH range of 6.6-7.5, as the pH value increased, the growth rate of SEC aggregates slowed down. When the pH was 6.6, the high molecular weight protein increased to 5.53% on the 20th day, exceeding the established upper limit quality standard of 5%. When the pH value was 6.7-7.5, the growth rate of SEC aggregates was below 5%.

[0337] Furthermore, the preparations F5-F11 were subjected to room temperature shaking study, with a shaking time of 72 hours and a shaking speed of 600 rpm. The results of the shaking experiment are shown in Table 15 below.

[0338] Table 15: SEC-HPLC aggregation change rate of preparations F5 to F11 placed under room temperature shaking conditions for 72 hours (days / %)

[0339] preparation 0h-SEC aggregates (%) Room temperature shaking for 72h-SEC aggregation (%) Change (%) F5 1.46 2.03 0.57 F6 1.44 1.96 0.52 F7 1.01 1.42 0.41 F8 0.89 1.33 0.44 F9 0.93 1.30 0.37 F10 0.88 1.22 0.34 F11 0.86 1.17 0.31

[0340] As shown in Table 15, after 72 hours of shaking, the SEC-aggregates of preparations F5-F11 only increased slightly, and the growth rate also slowed down with the increase of pH value. When the pH value of the preparation is 6.6-7.5, it can prevent the protein from forming more aggregates due to shaking.

[0341] In order to further evaluate the effects of different pH values ​​on the conformational stability and colloidal stability of proteins, the ThermControl module of NanoTemper's Prometheus NT.48 protein stability analyzer was used to test the preparations F5-F11 to obtain the denaturation temperature (Tm and Tonset) of the protein and the starting temperature (Tagg) of protein aggregation. The test results are shown in Table 16 below.

[0342] Table 16: Tagg and Tm value test results of preparations F5 to F11

[0343] preparation Tagg(℃) Tonset(℃) Tm1(℃) Tm2(℃) F5 62.3 54.9 62.0 70.9 F6 62.5 55.1 62.5 80.6 F7 64.4 56.4 63.7 80.2 F8 No aggregation detected 57.1 64.2 80.3 F9 No aggregation detected 57.0 64.5 80.4 F10 No aggregation detected 58.0 65.0 80.4 F11 No aggregation detected 58.5 65.5 80.3

[0344] As shown in Table 16, when the pH value is 6.6, Tonset, Tm and Tagg are lower than those of other groups, indicating poor stability. When the pH value is 6.7-7.5, as the pH increases, the Tonset and Tm1 values ​​increase, indicating better thermal stability. When the pH value is 7.0-7.5, no aggregation temperature Tagg value is detected, indicating good colloidal stability.

[0345] In summary, when the pH of the preparation is in the range of 6.7-7.5, the chemical stability and physical stability of the preparation are good.

[0346] 3. Investigation of injection solutions prepared with different types of surfactants

[0347] 3.1 Different surfactants were used to prepare low-concentration injection preparations A16 to A19. The final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 17. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the prescription amount in the following table. If necessary, the pH of the preparation is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally the injection water is added to the volume. After the preparation is completed, the drug solution is filtered in series with 0.45μm and 0.22μm PVDF filter membranes into a 1ml slender prefilled syringe, each filled with 0.8ml, and sealed with a rubber stopper. . Then, the high-temperature stability of preparations A16 to A19 was studied. The SEC-HPLC stability test results of 0 days and 10 days under high temperature 40°C are shown in Table 18, and the SEC-HPLC stability test results of 10 days under high temperature 40°C are shown in Table 19.

[0348] Table 17: Ratios of ingredients in formulations A16 to A19

[0349]

[0350] Table 18: Stability results of formulations A16 to A19 at high temperature 40°C

[0351]

[0352] Table 19: Change rate results of various indicators of formulations A16 to A19 at high temperature 40°C

[0353]

[0354] It can be seen from Tables 18 and 19 that different types of surfactants have little effect on SEC purity and aggregates, but have a greater effect on RP-HPLC-purity. Comprehensive analysis of the average daily decrease rate of RP-HPLC-purity (main peak + peak 4) and the average daily increase rate of RP-HPLC-purity (total related substances) shows that polysorbate 80 or polysorbate 20 is more effective as a stabilizer, but when poloxamer 188 is used as a stabilizer, the stability of the preparation is also within the acceptable standard range.

[0355] 3.2 Different surfactants were used to prepare high-concentration injection preparations F12 and F13. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 20. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the following table. If necessary, the pH of the solution is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally the volume is fixed to 15 ml with injection water. After the liquid is prepared, the liquid is filtered in series with 0.45 μm and 0.22 μm PVDF filter membranes into a 1 ml slender prefilled syringe, each with a volume of 0.5 ml, and sealed with a rubber stopper. Then, the preparations F12 and F13 were subjected to high temperature stability studies. The SEC-HPLC aggregation change rate (days / %) under high temperature conditions of 40°C for 28 days is shown in Table 21, and the appearance test results are shown in Table 22.

[0356] Table 20: Ratios of ingredients in formulations F12 and F13

[0357]

[0358] Table 21: SEC-HPLC aggregation change rate results of formulations F12-F13 placed at high temperature 40°C for 28 days

[0359] preparation Day 0-SEC Aggregates (%) High temperature 40℃28 days-SEC aggregates (%) Change rate (% / day) F12 1.49 2.76 0.05 F13 1.50 2.69 0.04

[0360] Table 22: Appearance test results of formulations F12 and F13 placed at 40°C for 28 days

[0361] preparation Day 0 - Appearance High temperature 40℃ 28 days - Appearance F12 Clear liquid with slight opalescence Clear liquid with slight opalescence F13 Clear liquid with slight opalescence Clear liquid with slight opalescence

[0362] It can be seen from Table 21 and Table 22 that the injection solutions prepared with polysorbate 80 and polysorbate 20 can slow down the growth of aggregates and prevent the formation of visible particles, thereby having a good stabilizing effect on the fusion protein.

[0363] 4. Investigation of injection solutions prepared with different types of osmotic pressure regulators

[0364] 4.1 Different types of osmotic pressure regulators were used to prepare low-concentration injection preparations A23 to A25. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically shown in Table 23. The preparation method of the injection is specifically shown in step 2.1 of the embodiment of this application. Then, the high-temperature stability of preparations A23 to A25 was studied. The SEC-HPLC stability test results of 0 days, 7 days and 14 days under high temperature conditions of 40°C are shown in Table 24.

[0365] Table 23: Ratios of ingredients in formulations A23 to A25

[0366]

[0367]

[0368] Table 24: Stability results of formulations A23 to A25 at high temperature 40°C

[0369]

[0370] As shown in Table 24, compared with the combination of sodium chloride, sodium chloride and sucrose, the RP-HPLC-purity (total related substances) of the preparation using mannitol increased the slowest, and less foreign matter was visible. Therefore, the preparation prepared using mannitol as an osmotic pressure regulator has better stability.

[0371] In addition, by testing the osmotic pressure values ​​of the low-concentration injection solutions under the dosage of mannitol, they were all equal to or close to the osmotic pressure of human plasma, among which the osmotic pressure of preparation A12 batch was 307mOsm / kg, the osmotic pressure of preparation A16 was 310mOsm / kg, the osmotic pressure of preparation A26 was 309mOsm / kg, and the osmotic pressure of preparation A29 batch was 325mOsm / kg.

[0372] 4.2 Different types of osmotic pressure regulators were used to prepare low-concentration injection preparations A34-A35. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically shown in Table 25. The preparation method of the injection is specifically shown in step 2.1 of the embodiment of the present application. Then, the high-temperature stability of preparations A34-A35 was studied. The SEC-HPLC stability test results of 0 days, 7 days and 14 days under high temperature conditions of 40°C are shown in Table 26.

[0373] Table 24: Ratios of ingredients in formulations A34 to A35

[0374]

[0375] Table 25: Stability results of formulations A34-A35 at high temperature 40°C

[0376]

[0377]

[0378] The stability of preparations prepared using sucrose and trehalose as osmotic pressure regulators is also better.

[0379] 4.3 Different osmotic pressure regulators were used to prepare high-concentration injection preparations F14 and F15. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 26. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the following table. If necessary, the pH of the solution is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally the volume is fixed to 9 ml with injection water. After the preparation is completed, the drug solution is filtered in series with 0.45 μm and 0.22 μm PVDF filter membranes into a 1 ml slender prefilled syringe, each filled with 0.5 ml, and sealed with a rubber stopper. Then, the preparations F14 and F15 were subjected to high temperature stability studies. The SEC-HPLC aggregation change rate (days / %) after being placed at high temperature of 40°C for 30 days is shown in Table 27, and the appearance test results after being placed at high temperature of 40°C for 14 days are shown in Table 28.

[0380] Table 26: Proportions of ingredients in formulations F14 and F15

[0381]

[0382] Table 27: SEC-HPLC aggregation change rate results of formulations F14-F15 placed at high temperature 40°C for 30 days

[0383] preparation Day 0-SEC Aggregates (%) High temperature 40℃30 days-SEC aggregates (%) Change rate (% / day) F14 1.99 3.43 0.05 F15 2.03 3.36 0.04

[0384] Table 28: Appearance test results of formulations F14 and F15 placed at 40°C for 14 days

[0385] preparation Day 0 - Appearance High temperature 40℃ 14 days - Appearance F14 Clear liquid with slight opalescence Clear liquid with slight opalescence F15 Clear liquid with slight opalescence Clear liquid with slight opalescence

[0386] In order to further evaluate the effects of different types of osmotic pressure regulators on the conformational stability and colloidal stability of proteins, the ThermControl module of NanoTemper's Prometheus NT.48 protein stability analyzer was used to test the preparations F14-F15 to obtain the denaturation temperature (Tm and Tonset) of the protein and the starting temperature (Tagg) of protein aggregation. The test results are shown in Table 29 below.

[0387] Table 29: Tagg and Tm value test results of preparations F14 to F15

[0388] preparation Tagg(℃) Tonset(℃) Tm1(℃) Tm2(℃) F14 No aggregation detected 56.32 63.42 80.34 F15 No aggregation detected 56.77 64.16 80.89

[0389] It can be seen from Tables 27 to 29 that mannitol or sucrose can maintain good colloidal stability and conformational stability of the dual-target fusion protein preparation, and can slow down the aggregation rate of the protein, prevent the protein from forming visible particles, and have a good stabilizing effect on the fusion protein.

[0390] 5. Investigation of injection solutions prepared with different histidine dosages

[0391] High-concentration injection preparations F16 to F18 were prepared using different amounts of histidine. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 30. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the following table. If necessary, the pH of the solution is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally the volume is fixed to 10 ml with injection water. After the preparation is completed, the drug solution is filtered in series with 0.45 μm and 0.22 μm PVDF filter membranes into a 1 ml slender pre-filled syringe, each filled with 0.5 ml, and sealed with a rubber stopper. Then the preparations F16 to F18 were subjected to a high temperature stability study, and the results of the SEC-HPLC aggregation change rate (days / %) at a high temperature of 40°C for 30 days are shown in Table 31.

[0392] Table 30: Proportions of the ingredients in formulations F16 to F18

[0393]

[0394] Table 31: SEC-HPLC aggregation change rate results of preparations F16 to F18 placed at high temperature 40°C for 30 days

[0395] preparation Day 0-SEC Aggregates (%) High temperature 40℃30 days-SEC aggregates (%) Change rate (% / day) F16 0.89 2.76 0.09 F17 1.00 2.97 0.10 F18 0.99 3.14 0.11

[0396] From the above, it can be seen that when the dosage of histidine is 10-20mM, the growth rate of SEC-aggregates of the fusion protein is slow, and it has a good stabilizing effect on the fusion protein.

[0397] 6. Investigation of injection solutions prepared with different dosages of polysorbate 80

[0398] 6.1 Low-concentration injection preparations A20-A22 and A32-A33 were prepared using different amounts of polysorbate 80. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically referred to Table 32. The preparation method of the injection is specifically referred to step 2.1 of the embodiment of the present application. Then, preparations A20-A22 and A32-A33 were subjected to a high temperature stability study. The SEC-HPLC stability test results of 0 days, 7 days, 14 days and 28 days under high temperature conditions of 40°C are shown in Table 33.

[0399] Table 32: Proportions of the ingredients in formulations A20-A22 and A32-A33

[0400]

[0401] Table 33: Stability results of formulations A20-A22 and A32-A33 at high temperature 40°C

[0402]

[0403]

[0404] Note: “ / ” means not detected.

[0405] It can be seen from Table 33 that when the dosage of polysorbate 80 is in the range of 0.1 mg / ml to 0.5 mg / ml, the stability of the injection is better.

[0406] 6.2 High-concentration injection preparations F19 to F25 were prepared using different amounts of polysorbate 80. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 34. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the following table. If necessary, the pH of the solution is adjusted to the corresponding value using 2% hydrochloric acid solution or 2% sodium hydroxide solution, and finally the volume is fixed to 15 ml with injection water. After the preparation is completed, the drug solution is filtered in series with 0.45 μm and 0.22 μm PVDF filter membranes into a 1 ml slender prefilled syringe, with a volume of 0.5 ml per syringe, and a rubber stopper is plugged and sealed. Then the preparations F19 to F25 were subjected to a high temperature stability study. The SEC-HPLC aggregation change rate (days / %) after being placed at a high temperature of 40°C for 28 days is shown in Table 35, the appearance test results are shown in Table 36, and the insoluble particle test results are shown in Table 37.

[0407] Table 34: Proportions of the ingredients in formulations F19 to F25

[0408]

[0409] Table 35: SEC-HPLC aggregation change rate results of formulations F19 to F25 placed at high temperature 40°C for 28 days

[0410] preparation Day 0-SEC Aggregates (%) High temperature 40℃28 days-SEC aggregates (%) Change rate (% / day) F19 1.19 2.31 0.04 F20 1.21 2.46 0.04 F21 1.21 2.34 0.04 F22 1.21 2.57 0.05 F23 1.33 3.82 0.09 F24 1.32 3.76 0.09 F25 1.33 3.86 0.09

[0411] Table 36: Appearance test results of formulations F19 to F25 placed at 40°C for 28 days

[0412] preparation Day 0 - Appearance High temperature 40℃ 28 days - Appearance F19 Clear liquid with slight opalescence A small amount of white particles appear F20 Clear liquid with slight opalescence A small amount of white particles appear F21 Clear liquid with slight opalescence Clear liquid with slight opalescence F22 Clear liquid with slight opalescence Clear liquid with slight opalescence F23 Clear liquid with slight opalescence Clear liquid with slight opalescence F24 Clear liquid with slight opalescence Clear liquid with slight opalescence F25 Clear liquid with slight opalescence Clear liquid with slight opalescence

[0413] Table 37: Results of insoluble particles test of formulations F19 to F25 placed at 40°C for 28 days

[0414]

[0415] From the above, it can be seen that when the dosage of polysorbate 80 is 0.4-1.3 mg / ml, the stability of the dual-target fusion protein can be maintained, the growth rate of protein aggregates is slow, the number of insoluble particles is small, the protein is prevented from forming visible particles and the formation of insoluble particles is reduced.

[0416] 7. Investigation of injection solutions prepared with different fusion protein concentrations

[0417] 7.1 Different fusion protein concentrations were used to prepare low-concentration injection preparations A26 to A31. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically shown in Table 38. The preparation method of the injection is specifically shown in step 2.1 of the embodiment of this application. Then, the high-temperature stability of preparations A26 to A31 was studied. The SEC-HPLC stability test results of 0 days, 7 days, 14 days and 28 days under high temperature 40°C are shown in Table 39. The high-temperature SEC-aggregate growth trend of preparations of different concentrations is shown in Table 39. Figure 1 , the high temperature RP-HPLC-purity (total related substances) growth trend of preparations with different concentrations is shown in Figure 2 .

[0418] Table 38: Ratios of ingredients in formulations A26 to A31

[0419]

[0420] Table 39: Stability results of formulations A26-A31 at high temperature 40°C

[0421]

[0422]

[0423] From Table 39 and Figures 1-2 It can be seen that there is no significant difference in the growth rate of RP-HPLC-purity (total related substances) of preparations with different concentrations, and the growth rate of SEC-purity (aggregate) (%) increases with the increase of preparation concentration.

[0424] The long-term stability of formulation A29 (17 mg / ml) and formulation A30 (20 mg / ml) at 2-8°C was further investigated. The test results are shown in Table 40.

[0425] Table 40: Stability results of formulations A29-A30 at 2-8°C

[0426]

[0427] Note: “ / ” means not tested.

[0428] As can be seen from Table 40, with the change of storage time, the RP-HPLC-purity (main peak + peak 4) of preparations of different concentrations did not change significantly, and the SEC-purity (aggregate) had a slight increasing trend, but was within the acceptable standard range (≤5%).

[0429] 7.2 Different fusion protein concentrations were used to prepare low-concentration injection preparations F26 to F29. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically referred to Table 41. The preparation method of the injection is specifically referred to step 2.1 of the embodiment of the present application. Then the preparations F26 to F29 were subjected to a high temperature stability study, and the SEC-HPLC stability test was placed at a high temperature of 40°C for 0 days, 7 days, 14 days and 28 days. The results showed that the SEC-purity (polymer) of the preparations of different concentrations had a slight increasing trend, but they were all within the acceptable standard range (≤5%). This example exemplifies the results of preparation F26, as shown in Tables 42 and 43.

[0430] Table 41: Proportions of the ingredients in formulations F26 to F29

[0431]

[0432]

[0433] Table 42: Results of the rate of change of SEC-HPLC aggregates and RP-HPLC main peak purity at high temperature of 40°C for 28 days

[0434] preparation Day 0-SEC Aggregates (%) High temperature 40℃28 days-SEC aggregates (%) Change rate (% / day) F26 0.61 2.65 0.07 preparation Day 0 - Main peak purity % (including peak 4) High temperature 40℃28 days-main peak purity% (including peak 4) Change rate (% / day) F26 84.04 68.90 0.54

[0435] Table 43: Appearance test results after being placed at 40°C for 28 days

[0436] preparation Day 0 - Appearance High temperature 40℃ 28 days - Appearance F26 Clear liquid with slight opalescence Clear liquid with slight opalescence

[0437] 8. Effect of EDTA sodium salt on the stability of preparations under different buffer systems

[0438] 8.1 Prepare low-concentration injection preparations A36-A37 containing EDTA in a citric acid system. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically shown in Table 44. The preparation method of the injection is specifically shown in step 2.1 of the embodiment of this application. Then, the high-temperature stability of preparations A36-A37 was studied. The SEC-HPLC stability test results of 0 days, 10 days, 14 days and 28 days under high temperature conditions of 40°C are shown in Table 45.

[0439] Table 44: Ratios of ingredients in formulations A36 to A37

[0440]

[0441] Table 45: Stability results of formulations A36-A37 at high temperature 40°C

[0442]

[0443]

[0444] 8.2 Preparation of high-concentration injection preparations F30-F31 containing EDTA under the histidine system, the final concentration of GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection and the types and final concentrations of each excipient are specifically referred to Table 46, and the preparation method of the injection is specifically referred to step 2.1 of the embodiment of the present application. Then the preparations F30-F31 were subjected to a high temperature stability study, and the SEC-HPLC stability test results of 0 days, 7 days, 14 days and 30 days under high temperature conditions of 40°C are shown in Table 47.

[0445] Table 46: Ratios of ingredients in formulations F30 to F31

[0446]

[0447] Table 47: Stability results of formulations F30-F31 at high temperature 40°C

[0448]

[0449] In summary, whether it is a citric acid system or a histidine buffer system, the stability of the preparation is better with or without the addition of EDTA sodium salt.

[0450] Example 2: Investigation of different fusion proteins

[0451] The amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 2 in this example is shown in SEQ ID NO: 2; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 3 in this example is shown in SEQ ID NO: 3; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 4 in this example is shown in SEQ ID NO: 4; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 5 in this example is shown in SEQ ID NO: 5; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 6 in this example is shown in SEQ ID NO: 6; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 7 in this example is shown in SEQ ID NO: 7; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 8 in this example is shown in SEQ ID NO: 8; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 9 in this example is shown in SEQ ID NO:9; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 10 in this embodiment is shown in SEQ ID NO:10; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 11 in this embodiment is shown in SEQ ID NO:11; the amino acid sequence of the GLP-1-Fc-FGF21 dual-target fusion protein 12 in this embodiment is shown in SEQ ID NO:12. Among them, GLP-1-Fc-FGF21 dual-target fusion proteins 2 to 11 (referred to as fusion proteins 2 to 11) are all prepared by conventional methods in the art.

[0452] 1. The type and final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection, as well as the type and final concentration of each excipient, are specifically referred to Table 48. The preparation method of the injection is specifically referred to step 2.1 of Example 1. Then, the stability of preparations A38 to A31 was studied.

[0453] Table 48: Ratios of ingredients in formulations A38 to A31

[0454]

[0455] 2. The type and final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (hereinafter referred to as fusion protein) in the injection, as well as the type and final concentration of each excipient, are specifically referred to Table 49. The preparation method of the injection is specifically referred to step 2.1 of Example 1. Then, the stability of preparations F32 to F53 was studied.

[0456] Table 49: Proportions of the ingredients in formulations F32 to F53

[0457]

[0458]

[0459] Example 3: Investigation of different preservative systems

[0460] Different preservative systems were used to prepare low-concentration injection preparations. The final concentration of the GLP-1-Fc-FGF21 dual-target fusion protein (abbreviated as fusion protein, i.e., API) in the injection and the types and final concentrations of each excipient are specifically shown in Table 50. The preparation method of the injection is as follows: the excipients and API are weighed and mixed according to the prescription amount in Table 1. If necessary, the pH of the preparation is adjusted to the corresponding value using a 2% hydrochloric acid solution or a 2% sodium hydroxide solution, and finally the injection water is added to the volume. After the liquid is prepared, the liquid is filtered in series with 0.45μm and 0.22μm PVDF filter membranes into a 1ml slender prefilled syringe, each filled with 1ml, and sealed with a rubber stopper. The preparations were then subjected to stability studies, and the results showed that each preparation showed good stability, and the SEC-HPLC aggregate change rate (day / %) was within the acceptable standard range, and no visible particles were formed.

[0461] Table 50: Ratio of ingredients in each preparation

[0462]

[0463]

[0464] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0465] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A composition, characterized in that include: GLP-1-Fc-FGF21 fusion protein and buffer.

2. The composition according to claim 1, characterized in that The GLP-1-Fc-FGF21 fusion protein comprises the amino acid sequence 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.

3. The composition according to claim 2, characterized in that The buffer comprises at least one of a histidine buffer, a citrate buffer, a phosphate buffer and a Tris buffer; Preferably, the buffer comprises histidine buffer or 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 in the composition is 10-20 mM.

4. The composition according to claim 1, characterized in that The composition further comprises at least one of an osmotic pressure regulator, a surfactant, a preservative, and a metal chelator; Optionally, the osmotic pressure regulator includes 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 includes 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; Optionally, the preservative is 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; Optionally, the metal chelator is at least one of edetate disodium and edetate calcium sodium; Optionally, the concentration of the metal chelator in the composition is 0.01 to 1 mg / mL, preferably 0.05 to 0.5 mg / mL.

5. The composition according to any one of claims 1 to 4, characterized in that 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, 8 to 20 mM, more preferably 10 to 20 mM, 8 to 12 mM; Optionally, the pH value 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, 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.

6. The composition according to any one of claims 1 to 4, characterized in that 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, 8 to 20 mM, more preferably 10 to 20 mM, 8 to 12 mM; Optionally, the pH value 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, and 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.

7. The composition according to any one of claims 1 to 4, characterized in that The dosage form of the composition is a liquid preparation, preferably an injection.

8. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol and optionally edetate disodium or edetate sodium calcium, and polysorbate 80 or polysorbate 20; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 mg / mL; The concentration of the histidine buffer in the injection solution is 8-25 mM, preferably 10-20 mM; The concentration of mannitol in the injection is 35-45 mg / mL; The concentration of the polysorbate 80 in the injection is 0.4 to 1.5 mg / mL, or 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; The concentration of the metal chelator in the injection is 0.05-0.5 mg / mL; The pH value of the injection solution is 6.9 to 7.5; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

9. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, Histidine buffer, Sucrose or trehalose, Polysorbate 80 or polysorbate 20, and Optionally edetate disodium or edetate calcium sodium; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 mg / mL; The concentration of the histidine 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 the polysorbate 80 in the injection is 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; The pH value of the injection solution is 6.9 to 7.5; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

10. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, citrate buffer, sucrose or trehalose, and optionally edetate disodium or edetate sodium calcium, and polysorbate 80 or polysorbate 20; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 25 to 50 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 the polysorbate 80 in the injection is 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; The pH value of the injection solution is 6.9 to 7.5; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

11. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, and optionally edetate disodium or edetate sodium calcium, and polysorbate 80 or polysorbate 20; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 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, 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 injection is 35-45 mg / mL; The concentration of the polysorbate 80 in the injection is 0.4 to 1.5 mg / mL, or 0.4 to 1.3 mg / mL, or the concentration of the polysorbate 20 in the injection is 0.1 to 0.5 mg / mL; The concentration of the metal chelator in the injection is 0.05-0.5 mg / mL; The pH value of the injection solution is 6.9 to 7.5; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

12. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, citrate buffer, mannitol, polysorbate 80, and optionally edetate disodium or edetate calcium sodium; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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, preferably 10 to 15 mM; The concentration of mannitol in the injection is 40-50 mg / mL; The concentration of polysorbate 80 in the injection is 0.1 to 1.5 mg / mL; The concentration of the metal chelator in the injection is 0.05-0.5 mg / mL; The pH value of the injection is 6.4 to 8.0; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

13. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, citrate buffer and polysorbate 80, and sucrose or trehalose, and optionally edetate disodium or edetate calcium sodium; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 is 5 to 20 mM, preferably 8 to 20 mM, more preferably 10 to 15 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 is 0.1 to 1.5 mg / mL; The pH value of the injection is 6.4 to 8.0; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

14. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, histidine buffer, mannitol, polysorbate 80, and optionally edetate disodium or edetate calcium sodium; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 5-30 mM, preferably 10-20 mM; The concentration of mannitol in the injection is 40-50 mg / mL; The concentration of polysorbate 80 in the injection is 0.1 to 1.5 mg / mL; The concentration of the metal chelator in the injection is 0.05-0.5 mg / mL; The pH value of the injection is 6.4 to 8.0; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

15. An injection, characterized in that: include: GLP-1-Fc-FGF21 fusion protein, histidine buffer and polysorbate 80, and sucrose or trehalose, and optionally edetate disodium or edetate calcium sodium; The GLP-1-Fc-FGF21 fusion protein comprises an amino acid sequence as 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 5-30 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 is 0.1 to 1.5 mg / mL; The pH value of the injection is 6.4 to 8.0; Preferably, the injection does not contain edetate disodium or edetate calcium sodium.

16. The injection according to any one of claims 8 to 15, characterized in that: Also contains preservatives.

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

18. Use of the composition according to any one of claims 1 to 7 or the injection according to any one of claims 8 to 17 in the preparation of a medicament for treating and / or preventing metabolic diseases; Optionally, the metabolic diseases include diabetes, hyperlipidemia, obesity, and diseases associated with fatty liver; Optionally, the fatty liver-related diseases include non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, liver fibrosis and liver cirrhosis.

19. An injection device or container, characterized in that: include: The composition according to any one of claims 1 to 7 or the injection according to any one of claims 8 to 17; Optionally, the injection device or container comprises at least one selected from a container bottle, a vial, a cartridge and a syringe; Preferably, the syringe comprises at least one selected from an autoinjector and a pre-filled syringe.

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