Pharmaceutical combinations of PCSK9 inhibitors and GLP-1 receptor agonists

Abstract

The invention relates to a pharmaceutical composition. The pharmaceutical composition comprises a PCSK9 inhibitor and a GLP-1 receptor agonist, the invention also relates to application of the medicine composition in preparation of medicines for treating hyperglycemia, diabetes, obesity and / or cholesterol-related diseases.

Description

Technical Field

[0001] The present invention relates to a drug combination, comprising a) a PCSK9 inhibitor and b) a GLP-1 receptor agonist; and also relates to use of the drug combination in preparing a drug for treating hyperglycemia, diabetes, obesity and / or cholesterol-related diseases. Background Art

[0002] High blood sugar and high blood lipids are currently one of the most serious public health problems faced by all countries, and are particularly evident in middle-aged and elderly people.

[0003] Cholesterol is an essential lipid substance for human life activities. Increased cholesterol levels in plasma can cause atherosclerotic cardiovascular diseases and greatly increase the risk of myocardial infarction and stroke. 60% to 70% of plasma cholesterol in the human body is transported by low-density lipoprotein (LDL), and about 75% of LDL in plasma is endocytosed into liver cells through the low-density lipoprotein receptor (LDLR) of liver cells and cleared, and the remaining 25% is absorbed and utilized by peripheral tissues and organs. The LDLR-mediated LDL endocytosis pathway is considered to be the most important and efficient way for the body to clear plasma LDL. The protein drugs currently on the market or under development for alleviating or treating cholesterol-related symptoms mainly include PCSK9 (proprotein convertase subtilisekexin9, PCSK9) inhibitors, ANGPTL3 (angiopoietin-like 3) inhibitors, ANGPTL4 / 8 (angiopoietin-like protein 4 / 8, Angiopoietin-like protein 4 / 8) inhibitors, Apoc3 (apolipoprotein C3, apolipoprotein C3) inhibitors and apolipoprotein (a) (lipoprotein a) inhibitors.

[0004] PCSK9 (proprotein convertase subtilisekexin 9, PCSK9) protein mediates the degradation of LDL receptors by binding to the LDL receptors on the surface of liver cells. At this time, the amount of LDL that can be mediated by LDL receptors to enter liver cells for degradation is reduced, resulting in an increase in the LDL level in plasma. PCSK9 monoclonal antibody drugs are currently very good lipid-lowering drugs. They mediate PCSK9 degradation by binding to PCSK9, thereby increasing the level of LDL receptors on the surface of liver cells to reduce the level of LDL in plasma and achieve the effect of lowering blood lipids. The PCSK9 antibodies currently on the market mainly include Alirocumab from Sanofi and Evolocumab from Amgen. Others in the clinical stage include Lodelcizumab (Norvatis), Ralpancizumab (Pfizer), Bococizumab (Pfizer), LY3015014 (Eli Lilly), LIB-003 (Adenexus Therapeutics Inc), SHR-1209 (Suzhou Shengdiya Biomedical Co Ltd), AK-102 (Akeso BiopHarma Inc), JS-002 (Shanghai Junshi Biosciences Co Ltd), SAL-003 (Shenzhen Salubris pHarmaceuticals Co Ltd), AK-102 (Akeeso BiopHarma Inc), ATH-06 (AFFiRiS), etc.

[0005] Currently, the polypeptide drugs used to lower blood sugar mainly include GLP-1 (glucagon-like peptide-1, Glucagon-Like Peptide-1, GLP-1) receptor agonists, GIP (gastric inhibitory polypeptide, GIP) receptor agonists, GCGR (glucagon receptor) receptor agonists, FGF21 (Fibroblast growth factor 21) agonists, and agonists targeting any two or three targets of GLP-1, GIP, GCGR or FGF21.

[0006] GLP-1 (Glucagon-Like Peptide-1, GLP-1) is secreted by gastrointestinal mucosal L cells in response to meals. GLP-1 mainly acts through the GLP-1 receptor (GLP-1R) expressed on pancreatic β cells and δ cells (D cells). When GLP-1 binds to GLP-1R, it can mediate the secretion of somatostatin (SMS) from pancreatic δ cells. SMS in turn inhibits the secretion of glucagon from pancreatic α cells through somatostatin 2 receptor (SSTR2). On the other hand, GLP-1 mediates the increase in insulin synthesis and secretion through the GLP-1R on β cells, and plays a role in lowering blood sugar through the simultaneous action of these two aspects. The GLP-1 receptor agonists currently on the market mainly include exenatide, belaglutide and liraglutide, and the long-acting GLP-1 receptor agonists include polyethylene glycol loxenatide, semaglutide, albiglutide, dulaglutide, etc.

[0007] According to the literature (see Cardiovascular safety and efficacy of the PCSK9 inhibitor evolocumab in patients with and without diabetes and the effect of evolocumab on glycaemia and risk of new-onset diabetes: a repespecified analysis of the FOURIER randomised controlled trial, Marc S Sabatine, Lawrence A Leiter, et al., Lancet Diabetes Endocrinol. Volume 5, Issue 12, December 2017, Pages 941-950), among patients with hyperlipidemia, about 40% of them also suffer from hyperglycemia; and among patients with hyperglycemia, about 40% of them still have low-density lipoprotein cholesterol (LDL-C) levels that are not up to standard after taking statins for lipid-lowering treatment. Therefore, the number of patients who need to control both blood lipids and blood sugar at the same time is very large.

[0008] In order to serve the above-mentioned people with high blood lipids and high blood sugar, MedImmune has developed a fusion protein of PCSK9 monoclonal antibody and GLP-1 peptide-like peptide, MEDI4166, with the aim of achieving the lipid-lowering effect of PCSK9 monoclonal antibody and the blood sugar-lowering effect of GLP-1 peptide-like peptide. However, when MEDI4166 entered Phase I clinical trials, MEDI4166 (maximum dose 400 mg) only showed the effect of lowering LDL-C, but no blood sugar-lowering effect was found. Therefore, it is urgent to develop a drug that can treat both hyperglycemia and hyperlipidemia. Summary of the invention

[0009] The present invention provides a drug combination comprising a PCSK9 inhibitor and GLP-1, which, while maintaining the effect of the PCSK9 inhibitor in preventing or treating cholesterol-related symptoms, unexpectedly also maintains the blood sugar lowering function of GLP-1 or has a good synergistic effect in terms of blood sugar lowering. The pharmaceutical composition of the present invention has excellent effects on lowering blood sugar, reducing body weight and alleviating cholesterol-related symptoms.

[0010] The first aspect of the present invention provides a pharmaceutical combination comprising:

[0011] a) a protein drug with a content of 50 mg / ml to 200 mg / ml for alleviating or treating cholesterol-related symptoms, preferably, the protein drug is selected from PCSK9 inhibitors, ANGPTL3 inhibitors, ANGPTL4 / 8 inhibitors, Apoc3 inhibitors and apolipoprotein (a) inhibitors, and more preferably, the protein drug is a PCSK9 inhibitor; and

[0012] b) A hypoglycemic polypeptide drug with a content of 2 mg / ml-30 mg / ml. Preferably, the polypeptide drug is selected from GLP-1 receptor agonists, GIP receptor agonists, GCCR receptor agonists, FGF21 agonists, and agonists targeting any two or three targets of GLP-1, GIP, GCCR or FGF21. More preferably, the polypeptide drug is a GLP-1 receptor agonist.

[0013] Optionally, the content of the protein drug for alleviating or treating cholesterol-related symptoms is 60mg / ml-180mg / ml, more preferably 70mg / ml-160mg / ml, and further preferably 70mg / ml, 75mg / ml, 80mg / ml, 85mg / ml, 90mg / ml, 95mg / ml, 100mg / ml, 105mg / ml, 110mg / ml, 115mg / ml, 120mg / ml, 125mg / ml, 130mg / ml, 135mg / ml, 140mg / ml, 145mg / ml, 150mg / ml, 155mg / ml or 160mg / ml.

[0014] Optionally, the content of the hypoglycemic polypeptide drug is 2mg / ml-50mg / ml, more preferably 3mg / ml-40mg / ml, and further preferably 3mg / ml, 3.5mg / ml, 4mg / ml, 4.5mg / ml, 5mg / ml, 5mg / ml, 6mg / ml, 6.5mg / ml, 7mg / ml, 7.5mg / ml, 8mg / ml, 8.5mg / ml, 9mg / ml, 9.5mg / ml, 10mg / ml ml, 10.5mg / ml, 11mg / ml, 11.5mg / ml, 12mg / ml, 12.5mg / ml, 13mg / ml, 13.5mg / ml, 14mg / ml, 14.5mg / ml, 15 mg / ml, 15.5mg / ml, 16mg / ml, 16.5mg / ml, 17mg / ml, 17.5mg / ml, 18mg / ml, 18.5mg / ml, 19mg / ml, 19.5mg / ml, 20mg / ml, 20.5mg / ml, 21mg / ml, 21.5mg / ml, 22mg / ml, 22.5mg / ml, 23mg / ml, 23.5mg / ml, 24mg / ml, 24.5mg / ml, 25mg / ml, 25.5mg / ml, 26mg / ml, 26.5mg / ml, 27mg / ml, 27.5mg / ml, 28mg / ml, 28.5mg / ml, 29mg / ml, 29.5m g / ml, 30mg / ml, 31.5mg / ml, 32mg / ml, 32.5mg / ml, 33mg / ml, 33.5mg / ml, 34mg / ml, 34.5mg / ml, 35mg / ml, 35 .5mg / ml, 36mg / ml, 36.5mg / ml, 37mg / ml, 37.5mg / ml, 38mg / ml, 38.5mg / ml, 39mg / ml, 39.5mg / ml, 40mg / ml.

[0015] Optionally, the PCSK9 inhibitor is an antibody or an antigen-binding fragment thereof that specifically binds to PCSK9.

[0016] Optionally, the PCSK9 inhibitor is selected from at least one of alirocumab, evolocumab, lodelcizumab, ralpancizumab, bococizumab, inusitumob, LY3015014, LIB-003, SHR-1209, JS-002, SAL-003 and ATH-06; preferably, the PCSK9 inhibitor is alirocumab or evolocumab.

[0017] Optionally, the hypoglycemic polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai or glutazumab.

[0018] Optionally, the combination comprises

[0019] a) The content is 60mg / ml-180mg / ml, more preferably 60mg / ml-180mg / ml, more preferably 70mg / ml-160mg / ml, and further preferably 70mg / ml, 75mg / ml, 80mg / ml, 85mg / ml, 90mg / ml, 95mg / ml, 100mg / ml, 105mg / ml, 110mg / ml, 115mg / ml, 120mg / ml, 125mg / ml, 130mg / ml, 1 35 mg / ml, 140 mg / ml, 145 mg / ml, 150 mg / ml, 155 mg / ml or 160 mg / ml of a PCSK9 inhibitor, wherein the PCSK9 inhibitor is selected from at least one of Alisikumab, Elotumab, Lodelcizumab, Ralpancizumab, Bococizumab, Inusinumab, LY3015014, LIB-003, SHR-1209, JS-002, SAL-003 and ATH-06.

[0020] b) the content is 2mg / ml-50mg / ml, more preferably 3mg / ml-40mg / ml, and further preferably 3mg / ml, 3.5mg / ml, 4mg / ml, 4.5mg / ml, 5mg / ml, 5mg / ml, 6mg / ml, 6.5mg / ml, 7mg / ml, 7.5mg / ml, 8mg / ml, 8.5mg / ml, 9mg / ml, 9.5mg / ml, 10mg / ml, 10.5mg / ml, 11mg / ml, 11.5m g / ml, 12mg / ml, 12.5mg / ml, 13mg / ml, 13.5mg / ml, 14mg / ml, 14.5mg / ml, 15mg / ml, 15.5mg / ml, 16mg / ml, 16.5mg / ml , 17mg / ml, 17.5mg / ml, 18mg / ml, 18.5mg / ml, 19mg / ml, 19.5mg / ml, 20mg / ml, 20.5mg / ml, 21mg / ml, 21.5mg / ml, 22mg / ml, 22.5mg / ml, 23mg / ml, 23.5mg / ml, 24mg / ml, 24.5mg / ml, 25mg / ml, 25.5mg / ml, 26mg / ml, 26.5mg / ml, 27mg / ml, 27.5mg / ml, 28mg / ml, 28.5mg / ml, 29mg / ml, 29.5mg / ml, 30mg / ml, 31.5mg / ml, 32mg / ml, 32.5mg / ml, 33mg / ml, 33.5mg / ml g / ml, 34mg / ml, 34.5mg / ml, 35mg / ml, 35.5mg / ml, 36mg / ml, 36.5mg / ml, 37mg / ml, 37.5mg / ml, 38mg / ml, 38.5mg / ml, 39mg / ml, 39.5mg / ml, 40mg / ml of GLP-1 receptor agonist, the hypoglycemic polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26-[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai or glutazumab.

[0021] The present invention also provides a drug combination, comprising:

[0022] a) Inusimab,

[0023] b) A blood sugar lowering polypeptide drug. Preferably, the polypeptide drug is selected from a GLP-1 receptor agonist, a GIP receptor agonist, a GCCR receptor agonist, an FGF21 agonist, and an agonist targeting any two or three targets of GLP-1, GIP, GCCR or FGF21. Further preferably, the polypeptide drug is a GLP-1 receptor agonist.

[0024] Optionally, the hypoglycemic polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai or glutazumab.

[0025] Optionally, the content of inusimab is 60mg / ml-180mg / ml, more preferably 70mg / ml-160mg / ml, and further preferably 70mg / ml, 75mg / ml, 80mg / ml, 85mg / ml, 90mg / ml, 95mg / ml, 100mg / ml, 105mg / ml, 110mg / ml, 115mg / ml, 120mg / ml, 125mg / ml, 130mg / ml, 135mg / ml, 140mg / ml, 145mg / ml, 150mg / ml, 155mg / ml or 160mg / ml.

[0026] Optionally, the content of the hypoglycemic polypeptide drug is 2mg / ml-50mg / ml, more preferably 3mg / ml-40mg / ml, and further preferably 3mg / ml, 3.5mg / ml, 4mg / ml, 4.5mg / ml, 5mg / ml, 5mg / ml, 6mg / ml, 6.5mg / ml, 7mg / ml, 7.5mg / ml, 8mg / ml, 8.5mg / ml, 9mg / ml, 9.5mg / ml, 10mg / ml ml, 10.5mg / ml, 11mg / ml, 11.5mg / ml, 12mg / ml, 12.5mg / ml, 13mg / ml, 13.5mg / ml, 14mg / ml, 14.5mg / ml, 15 mg / ml, 15.5mg / ml, 16mg / ml, 16.5mg / ml, 17mg / ml, 17.5mg / ml, 18mg / ml, 18.5mg / ml, 19mg / ml, 19.5mg / ml, 20mg / ml, 20.5mg / ml, 21mg / ml, 21.5mg / ml, 22mg / ml, 22.5mg / ml, 23mg / ml, 23.5mg / ml, 24mg / ml, 24.5mg / ml, 25mg / ml, 25.5mg / ml, 26mg / ml, 26.5mg / ml, 27mg / ml, 27.5mg / ml, 28mg / ml, 28.5mg / ml, 29mg / ml, 29.5m g / ml, 30mg / ml, 31.5mg / ml, 32mg / ml, 32.5mg / ml, 33mg / ml, 33.5mg / ml, 34mg / ml, 34.5mg / ml, 35mg / ml, 35 .5mg / ml, 36mg / ml, 36.5mg / ml, 37mg / ml, 37.5mg / ml, 38mg / ml, 38.5mg / ml, 39mg / ml, 39.5mg / ml, 40mg / ml.

[0027] Optionally, the combination comprises

[0028] a) 60 mg / ml-180 mg / ml, more preferably 70 mg / ml-160 mg / ml, even more preferably 70 mg / ml, 75 mg / ml, 80 mg / ml, 85 mg / ml, 90 mg / ml, 95 mg / ml, 100 mg / ml, 105 mg / ml, 110 mg / ml, 115 mg / ml, 120 mg / ml, 125 mg / ml, 130 mg / ml, 135 mg / ml, 140 mg / ml, 145 mg / ml, 150 mg / ml, 155 mg / ml or 160 mg / ml of inusimab,

[0029] b) 2mg / ml-50mg / ml, more preferably 3mg / ml-40mg / ml, further preferably 3mg / ml, 3.5mg / ml, 4mg / ml, 4.5mg / ml, 5mg / ml, 5mg / ml, 6mg / ml, 6.5mg / ml, 7mg / ml, 7.5mg / ml, 8mg / ml, 8.5mg / ml, 9mg / ml, 9.5mg / ml, 10mg / ml, 10.5mg / ml, 11mg / ml, 11.5mg / ml ml, 12mg / ml, 12.5mg / ml, 13mg / ml, 13.5mg / ml, 14mg / ml, 14.5mg / ml, 15mg / ml, 15.5mg / ml, 16mg / ml, 16.5mg / ml, 1 7mg / ml, 17.5mg / ml, 18mg / ml, 18.5mg / ml, 19mg / ml, 19.5mg / ml, 20mg / ml, 20.5mg / ml, 21mg / ml, 21.5mg / ml, 22mg / ml, 22.5mg / ml, 23mg / ml, 23.5mg / ml, 24mg / ml, 24.5mg / ml, 25mg / ml, 25.5mg / ml, 26mg / ml, 26.5mg / ml, 27mg / ml, 2 7.5mg / ml, 28mg / ml, 28.5mg / ml, 29mg / ml, 29.5mg / ml, 30mg / ml, 31.5mg / ml, 32mg / ml, 32.5mg / ml, 33mg / ml, 33.5m g / ml, 34mg / ml, 34.5mg / ml, 35mg / ml, 35.5mg / ml, 36mg / ml, 36.5mg / ml, 37mg / ml, 37.5mg / ml, 38mg / ml, 38.5mg / ml, 39mg / ml, 39.5mg / ml, 40mg / ml of GLP-1 receptor agonist The hypoglycemic polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26-[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai or glutazumab.

[0030] Optionally, the PCSK9 inhibitor and the GLP-1 receptor agonist are administered simultaneously or separately.

[0031] Optionally, the combination is in the form of a pharmaceutical composition or a kit.

[0032] The second aspect of the present invention provides a pharmaceutical composition, which comprises the drug combination described in any one of the first aspects.

[0033] Optionally, the pharmaceutical combination further comprises one or more pharmaceutically acceptable excipients; preferably, the pharmaceutically acceptable excipients are selected from at least one of a buffer, a stabilizer, a surfactant, and an isotonic agent.

[0034] Optionally, the buffer is selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, histidine, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer is selected from at least one of citric acid, sodium acetate and disodium hydrogen phosphate; preferably the buffer is a combination of citric acid and disodium hydrogen phosphate; and / or

[0035] The stabilizer is selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably, selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine and proline; preferably selected from at least one of histidine, proline and sucrose; and / or

[0036] The surfactant is selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188; and / or

[0037] The isotonic agent is selected from at least one of sodium chloride, propylene glycol, and glycerol, preferably propylene glycol.

[0038] Optionally, wherein

[0039] The content of the buffer in the combination is 0.05mM to about 100mM, preferably about 0.05mM to about 85mM, preferably about 0.05mM to about 82mM, preferably about 0.05mM to about 40mM, preferably about 0.1mM to about 30mM, preferably about 1mM to about 20mM, preferably about 5mM to about 15mM, preferably about 5mM to about 10mM; and / or

[0040] The stabilizer is present in the combination in an amount of about 0.1% w / v to about 15% w / v, preferably about 0.5% w / v to about 10% w / v, preferably about 2.5% w / v to about 10% w / v; and / or

[0041] The surfactant is present in the combination in an amount of about 0.001% w / v to about 10% w / v, preferably about 0.01% w / v to about 1% w / v; and / or

[0042] The pH of the pharmaceutical combination is about 4.0 to about 8.0, preferably about 4.5 to about 7.5, preferably about 4.5 to about 7.0, preferably about 4.5 to about 6.5, preferably about 5.6 to about 6.2, preferably about 4.5 to about 5.5, preferably about 5.0 to about 5.5.

[0043] The present invention also provides a pharmaceutical composition, comprising:

[0044] The drug combination of the first aspect;

[0045] A buffer, wherein the buffer is selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer is selected from at least one of citric acid, sodium acetate, and disodium hydrogen phosphate, preferably the buffer is a combination of citric acid and disodium hydrogen phosphate;

[0046] A stabilizer, the stabilizer is selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably, selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine, histidine and proline; preferably, the stabilizer is selected from at least one of histidine, proline and sucrose; and

[0047] A surfactant, wherein the surfactant is selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188.

[0048] Optionally, the pH of the pharmaceutical composition is about 4.0-about 8.0, preferably about 4.5-about 7.5, preferably about 4.5-about 7.0, preferably about 4.5-about 6.5, preferably about 4.5-about 6.2, preferably about 5.6-about 6.2, preferably about 6.0-about 6.2, preferably about 4.5-about 5.5, preferably about 5.0-about 5.5.

[0049] The third aspect of the present invention provides a freeze-dried pharmaceutical preparation, which is prepared by the pharmaceutical combination or pharmaceutical composition according to claims 1-20.

[0050] The third aspect of the present invention provides a kit, the kit comprising:

[0051] (1) A first therapeutic agent: comprising the GLP-1 receptor agonist as defined in the first aspect;

[0052] (2) The second therapeutic agent: comprising the PCSK9 inhibitor defined in the first aspect.

[0053] Optionally, the hypoglycemic polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai or glutazumab.

[0054] Optionally, the PCSK9 inhibitor is alirocumab, evolocumab or inusumab.

[0055] Optionally, the first therapeutic agent is a lyophilized powder preparation.

[0056] Optionally, the second therapeutic agent further comprises:

[0057] A buffer, wherein the buffer is selected from at least one of citric acid monohydrate, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffer, and preferably the buffer is selected from at least one of citric acid monohydrate, sodium acetate, and disodium hydrogen phosphate;

[0058] A stabilizer, the stabilizer is selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably, selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine, histidine and proline; preferably, the stabilizer is selected from at least one of histidine, proline and sucrose; and

[0059] A surfactant, wherein the surfactant is selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188.

[0060] The fourth aspect of the present invention provides use of the drug combination, composition or kit described in the first aspect, the second aspect or the third aspect in the preparation of a drug for treating hyperglycemia, diabetes, obesity and / or cholesterol-related diseases.

[0061] The present invention also provides the drug combination, composition or kit described in the first aspect, the second aspect and the third aspect, which is used to treat hyperglycemia, diabetes, obesity and / or cholesterol-related diseases.

[0062] The present invention also provides a method for treating hyperglycemia, diabetes, obesity and / or cholesterol-related diseases, comprising administering a therapeutically effective amount of the drug combination, composition or kit described in the first aspect, the second aspect, or the third aspect.

[0063] definition:

[0064] Certain terms used herein are described below. It should also be understood that, unless otherwise defined, the terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. It should also be understood that the terms used herein are only used for the purpose of describing specific embodiments and are not intended to be limiting, as the scope of the present invention is limited only by the appended claims.

[0065] In this application, when referring to a specifically recited numerical value or numerical range, the terms "about" or "approximately" as used herein generally mean within 20%, preferably within 10%, and more preferably within 5% of the given value or range.

[0066] In the present application, the term "w / v" is a concentration unit, which means grams per milliliter (g / ml).

[0067] In the present application, the term "pharmaceutical combination" refers to a fixed combination, or a non-fixed combination, or a kit in which two or more therapeutic agents can be administered simultaneously or separately in the form of one dosage unit.

[0068] In this application, the term "fixed combination" or "fixed dose" refers to a single dosage form formulated to deliver to a patient an amount of two therapeutic agents that are therapeutically effective in combination for treating a disease. A single dosage form is designed to deliver an amount of each agent together with any pharmaceutically acceptable carrier or excipient, i.e., both agents together, in one dosage form, for simultaneous administration to the patient to be treated.

[0069] In the present application, the term "non-fixed combination" means that the active ingredients, such as the GLP-1 receptor agonist and the PCSK9 inhibitor, are administered to a patient as separate entities simultaneously, concurrently or sequentially but with no specific time limits.

[0070] In the present application, the term "kit" means a set of products or a combination product package, that is, the first therapeutic agent GLP-1 receptor agonist and the second therapeutic agent PCSK9 inhibitor of the present invention are sold in the same packaging form, for example, in a specific embodiment, it contains a container containing a therapeutic agent (the container includes, for example, a bottle, a vial, a syringe, an injection pen, a blister pack, etc.), and it may also contain instructions for the simultaneous, sequential, separate or mixed administration of the two therapeutic agents, such as a label or instruction sheet, and it may also contain other substances desired based on commercial and user positions, including diluents, needles, syringes, etc. In another embodiment, the kit may include a first container containing a first therapeutic agent GLP-1 receptor agonist, and a second container containing a second therapeutic agent PCSK9 inhibitor; in another embodiment, the kit may include a two-chamber or multi-chamber container, and the first therapeutic agent GLP-1 receptor agonist and the second therapeutic agent PCSK9 inhibitor are separately contained in different cavities.

[0071] In the present application, a "pharmaceutical composition" or "preparation" comprises at least one active ingredient, such as a) a GLP-1 receptor agonist, and / or b) a PCSK9 inhibitor, and a pharmaceutically acceptable carrier, diluent or excipient.

[0072] As used herein, the term "excipient" refers to any non-therapeutic substance added to a formulation to provide, for example, desired consistency, viscosity, or stability.

[0073] The term "antibody and antigen-binding portion thereof" herein is used in the broadest sense and includes various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

[0074] In the present application, "antibody" generally refers to an immunoglobulin molecule comprising four polypeptide chains, i.e., two heavy (H) chains and two light (L) chains, interconnected by disulfide bonds, and multimers thereof (e.g., IgM); however, immunoglobulin molecules consisting only of heavy chains (i.e., lacking light chains) are also included in the definition of the term "antibody". Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains: CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The VH and VL regions can be further subdivided into hypervariable regions, called complementarity determining regions (CDRs), which alternate with more conserved regions called framework regions (FRs). Each VH and VL contains 3 CDRs and 4 FRs, arranged in the following order from amino-terminus to carboxyl-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0075] As used herein, the term "antigen-binding portion" or "antigen-binding fragment" of an antibody (or simply "antibody portion" or "antibody fragment") refers to one or more fragments of an antibody that retain the ability to specifically bind to PCSK9. As used herein, the term "antigen-binding portion" or "antibody portion" or "antibody fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (such as RSV-F protein). It has been demonstrated that the antigen-binding function of an antibody can be performed by certain fragments of a full-length antibody. The binding fragments encompassed by the term "antigen-binding portion" of an antibody include (i) a Fab fragment, i.e., a monovalent fragment consisting of VL, VH, CL1, and CH1 domains; (ii) a F(ab')2 fragment, i.e., a bivalent fragment consisting of two F(ab)' fragments linked by a disulfide bond in the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the single-armed VL and VH domains of an antibody; (v) a dAb fragment consisting of the VH domain; and (vi) CDRs. In addition, although the two domains VL and VH of the Fv fragment are encoded by different genes, they can be connected together by a synthetic linker through a recombinant method to form a single connected chain, wherein the VL and VH regions are paired to form a monovalent molecule (called single-chain Fv (scFv)). Such single-chain antibodies are also included in the term "antigen-binding portion" of the antibody. Other forms of single-chain antibodies, such as bispecific antibodies, are also included.

[0076] In this application, the term "specific binding" and the like refer to the formation of a relatively stable complex between an antibody or its antigen-binding fragment and an antigen under physiological conditions. Specific binding can be achieved by at least about 1x10 -6The method for determining whether two molecules specifically bind is well known in the art and includes, for example, equilibrium dialysis, surface plasmonics, and the like.

[0077] GLP-1 analogs and GLP-1 derivatives

[0078] The term "GLP-1 analogue" or "analogue of GLP-1" as used herein refers to a peptide or compound that is a variant of human glucagon-like peptide-1 (GLP-1 (7-37)), wherein one or more amino acid residues of GLP-1 (7-37) are replaced, and / or one or more amino acid residues are deleted, and / or one or more amino acid residues are added. Specifically, the sequence of GLP-1 (7-37) is shown in SEQ ID NO: 1 in the sequence listing. The peptide having the sequence shown in SEQ ID NO: 1 may also be referred to as "native" GLP-1 or "native" GLP-1 (7-37).

[0079] In the sequence listing, the first amino acid residue (histidine) of SEQ ID NO: 1 is numbered 1. However, hereinafter, according to the established practice in the art, the histidine residue is numbered 7, and the amino acid residues thereafter are numbered accordingly, ending with glycine 37. Therefore, generally, the amino acid residue numbering or position numbering of the GLP-1 (7-37) sequence referred to herein is a sequence starting with His at position 7 and ending with Gly at position 37.

[0080] [Gly8, Arg34]GLP-1-(7-37) peptide is a GLP-1 analog having Gly and Arg at positions corresponding to position 8 and position 34 of GLP-1 (7-37) (SEQ ID NO: 1), respectively.

[0081] [Arg34]GLP-1-(7-37) peptide is a GLP-1 analog having Arg at a position corresponding to position 34 of GLP-1(7-37) (SEQ ID NO: 1). Specifically, the amino acid sequences of [Gly8, Arg34]GLP-1-(7-37) peptide and [Arg34]GLP-1-(7-37) peptide are shown in SEQ ID NO: 2 and SEQ ID NO: 3 in the sequence listing, respectively.

[0082] In the context of a GLP-1 peptide or analog thereof, the term "derivative" as used herein refers to a chemically modified GLP-1 peptide or analog wherein one or more substituents have been covalently attached to the peptide. The substituents may also be referred to as side chains.

[0083] Unless otherwise stated, when reference is made to acylation with a lysine residue, this is understood to be with the epsilon-amino group thereof.

[0084] The GLP-1 derivatives of formula (B) of the present invention may exist in different stereoisomeric forms, which have the same molecular formula and linked atomic sequence, but differ only in the three-dimensional orientation of their atomic space. Unless otherwise specified, the present invention relates to all stereoisomeric forms of the claimed derivatives.

[0085] The term "peptide" when applied to, for example, the GLP-1 analogs of the present invention, refers to a compound comprising a series of amino acids interconnected by amide (or peptide) bonds.

[0086] In a specific embodiment, the peptide is largely or mainly composed of amino acids linked to each other by amide bonds (e.g., at least 50%, 60%, 70%, 80% or at least 90% of the molar mass). In another specific embodiment, the peptide is composed of amino acids linked to each other by peptide bonds.

[0087] Amino acids are molecules containing an amino group and a carboxylic acid group, optionally with one or more additional groups, often called side chains.

[0088] The term "amino acid" includes proteinogenic amino acids (encoded by the genetic code, including natural amino acids and standard amino acids), as well as non-proteinogenic (not found in proteins, and / or not encoded in the standard genetic code), and synthetic amino acids. Non-proteinogenic amino acids are parts that can be incorporated into peptides through peptide bonds, but are not proteinogenic amino acids. Synthetic non-proteinogenic amino acids include amino acids produced by chemical synthesis, i.e., D-isomers of amino acids encoded by the genetic code, such as D-alanine and D-leucine, Aib (α-aminoisobutyric acid), Abu (α-aminobutyric acid), 3-aminomethylbenzoic acid, anthranilic acid, deamino-histidine, β-analogs of amino acids such as β-alanine, etc., D-histidine, deamino-histidine, 2-amino-histidine, β-hydroxy-histidine, and homohistidine, etc.

[0089] Non-limiting examples of amino acids not encoded by the genetic code are γ-carboxyglutamate, ornithine, D-alanine, D-glutamine and phosphoserine. Non-limiting examples of synthetic amino acids are the D-isomers of amino acids, such as D-alanine and D-leucine, Aib (α-aminoisobutyric acid), β-alanine and des-amino-histidine (desH, alternative name imidazole propionic acid, abbreviated Imp).

[0090] In the following, all amino acids not indicating the optical isomer are understood to refer to the L-isomer (unless otherwise specified).

[0091] The preparation methods of the peptides of GLP-1 (7-37) and GLP-1 analogs of the present invention are well known in the art. For example, the GLP-1 peptide portion (or fragment thereof) of the derivative of the present invention and the GLP-1 analogs of the present invention can be produced by classical peptide synthesis, such as solid phase peptide synthesis using t-Boc or Fmoc chemistry or other well-established techniques, see, for example, Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999, Florencio Zaragoza "Organic Synthesis on solid Phase", Wiley-VCH Verlag GmbH, 2000, and "Fmoc Solid Phase Peptide Synthesis" edited by WC Chan and PD White, Oxford University Press, 2000.

[0092] In one embodiment, the complete GLP-1 analogs of the present invention, such as [Gly8, Arg34]GLP-1-(7-37) peptides, can be produced by recombinant methods, i.e., by culturing host cells that contain a DNA sequence encoding the analog and are capable of expressing the peptide in a suitable nutrient medium under conditions that allow expression of the peptide. Non-limiting examples of host cells suitable for expressing these peptides are: Escherichia coli, Saccharomyces cerevisiae, and mammalian BHK or CHO cell lines. In some embodiments, this fully recombinant fermentation step of the production process is desirable, for example, for production economics considerations.

[0093] The fusion protein inclusion body containing the GPL-1 compound main chain is denatured and renatured to obtain a fusion protein containing the correct conformation, and a series of treatments such as enzyme digestion, precipitation, centrifugation, etc. are performed to obtain a GLP-1 compound main chain with a high content. After purification by ion exchange chromatography, a GLP-1 compound main chain with a high purity is obtained.

[0094] The term "amino acid residue" encompasses amino acids from which hydrogen atoms have been removed from amino groups and / or hydroxyl groups have been removed from carboxyl groups and / or hydrogen atoms have been removed from sulfhydryl groups. Inaccurately, amino acid residues may be referred to as amino acids.

[0095] Unless otherwise specified, all amino acids mentioned herein are L-amino acids.

[0096] The term alkylene glycol comprises oligo- and polyalkylene glycol moieties and monoalkylene glycol moieties.Monoalkylene glycol and polyalkylene glycol comprise for example based on mono- and polyethylene glycol, based on mono- and polypropylene glycol and based on mono- and polybutylene glycol chains, i.e. based on repeating units-CH2CH2O-,-CH2CH2CH2O- or-CH2CH2CH2CH2O- chains.Alkylene glycol moieties can be monodisperse (with well-defined length / molecular weight) and polydisperse (with less well-defined length / mean molecular weight).Monoalkylene glycol moieties include-OCH2CH2O-,-OCH2CH2CH2O- or-OCH2CH2CH2CH2O- comprising different groups at each end.

[0097] The term "fatty acid" includes straight or branched chain aliphatic carboxylic acids having at least two carbon atoms and being saturated or unsaturated. Non-limiting examples of fatty acids are, for example, myristic acid, palmitic acid, stearic acid and eicosanoic acid.

[0098] Herein, the term "aliphatic diacid" includes straight or branched aliphatic dicarboxylic acids, which have at least two carbon atoms and are saturated or unsaturated. Non-limiting examples of aliphatic diacids are adipic acid, suberic acid, sebacic acid, dodecanedioic acid, tetradecanedioic acid, hexadecanedioic acid, heptadecanedioic acid, octadecanedioic acid, eicosanedioic acid, docosanedioic acid, and tetracosanedioic acid.

[0099] In this article, the naming of GLP-1 compounds is carried out according to the following principles: the name is given according to the mutation and modification (such as acylation) relative to the native GLP-1 (7-37). For the naming of the acyl part, it is named according to the IUPAC nomenclature and in other cases according to the peptide nomenclature. For example, the following acyl part is named:

[0100]

[0101] For example, it can be named "eicosanedioyl-γGlu-OEG-OEG", "eicosanedioyl-γGlu-2xOEG" or "eicosanedioyl-gGlu-2xOEG", "19-carboxynonadecanoyl-γGlu-2xOEG" or "19-carboxynonadecanoyl-γGlu-OEG-OEG", where OEG represents the abbreviation of the group -NH(CH2)2O(CH2)2OCH2CO- (i.e., 2-[2-(2-aminoethoxy)ethoxy]acetyl), and γGlu (and gGlu) is a shorthand representation of the amino acid γglutamic acid in the L configuration. Alternatively, the acyl moiety can be named according to the IUPAC nomenclature (OpenEye, IUPAC format). According to this nomenclature, the above-mentioned acyl moieties of the present invention are referred to as the following names: “[2-[2-[2-[2-[2-[2-[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-amino]-ethoxy]-ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]” or “[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl]”.

[0102] In this application, the term "synergistic effect" can be determined by the gold average Q value, where the Q value is calculated by the following formula: Q = E a+b / (E a +E b -E a ×E b );E a+b It indicates the decrease rate of the detection index when two drugs are used in combination or mixed, E a 、E b It indicates the decline rate of each test index when the two drugs are used alone. If Q is between 0.85 and 1.15, it means that the two drugs have an additive effect; if Q>1.15, the two drugs have a synergistic effect; and if Q<0.85, the two drugs have an antagonistic effect.

[0103] "Dialysis exchange buffer" uses a semipermeable membrane to wrap the protein solution and immerse it in the dialysis buffer. Since the concentration of the buffer is lower than the concentration inside the membrane, material exchange will occur on the semipermeable membrane; but proteins cannot pass through the semipermeable membrane, only small molecules such as salt ions can pass through.

[0104] The drug combination / drug composition provided by the present invention has the following advantages:

[0105] 1) The drug combination / drug composition / lyophilized preparation of the present invention maintains the effect of PCSK9 inhibitor in lowering blood lipids, and maintains the function of GLP-1 in lowering blood glucose or unexpectedly exhibits synergistic effects.

[0106] 2) The drug combination / composition / lyophilized preparation of the present invention fills the gap in the treatment of patients suffering from both hyperglycemia and cholesterol-related symptoms.

[0107] 3) The drug combination / composition / lyophilized preparation of the present invention has no significant difference in pharmacokinetic properties compared with a single drug and has good pharmacokinetic properties.

[0108] 4) The pharmaceutical combination / composition / lyophilized preparation of the present invention has good stability, and each component thereof meets the internal control quality standard after being placed for several months. BRIEF DESCRIPTION OF THE DRAWINGS

[0109] Figure 1 Kinetic affinity fitting curve of AK102 and PCSK9 in AK102 monoclonal antibody preparation

[0110] Figure 2 Kinetic affinity fitting curve of AK102 and PCSK9 in AK102 monoclonal antibody + compound 1 dry powder formulation

[0111] Figure 3 Kinetic affinity fitting curve of AK102 and PCSK9 in the liquid formulation of AK102 monoclonal antibody + compound 1

[0112] Figure 4 Effect of AK102 on LDL endocytosis in HepG2 cells DETAILED DESCRIPTION

[0113] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0114] Abbreviations

[0115] Na2HPO4 is disodium hydrogen phosphate;

[0116] NaOH is sodium hydroxide;

[0117] OEG is the amino acid residue -NH(CH2)2O(CH2)2OCH2CO-;

[0118] OSu is succinimidyl-1-yloxy-2,5-dioxo-pyrrolidin-1-yloxy;

[0119] OtBu is tert-butyloxy;

[0120] HCl is hydrogen chloride;

[0121] γGlu or gGlu is γL-glutamyl;

[0122] NHS is N-hydroxysuccinimide;

[0123] DCC is dicyclohexylcarbodiimide;

[0124] AEEA is 2-(2-(2-aminoethoxy)ethoxy)acetic acid;

[0125] OH is hydroxide;

[0126] Gly is glycine;

[0127] Arg is arginine;

[0128] TFA is trifluoroacetic acid;

[0129] SEC-HPLC is size exclusion high performance liquid chromatography (Size Exclusion Chromatography-High Performance Liquid Chromatography);

[0130] CEX-HPLC is cation exchange high performance liquid chromatography (Cation Exchange Chromatography-High Performance Liquid Chromatography);

[0131] CE-SDS is capillary gel electrophoresis sodium dodecyl sulfate;

[0132] RPC is Reversed-Phase Chromatography.

[0133] Table 1: Materials and Reagents

[0134]

[0135] Example 1 Preparation of GLP-1 receptor agonists

[0136] Preparation of the title compound: N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide (Compound 1)

[0137]

[0138] 1. N-ε 26 Preparation of -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide

[0139] [Gly8, Arg34]GLP-1-(7-37) peptide was prepared by a general protein recombinant expression method (for specific methods, see Molecular Cloning: A Laboratory Manual (Fourth Edition), Michael R. Green, Cold Spring Harbor Press, 2012). [Gly8, Arg34]GLP-1-(7-37) peptide (5 g, 1.48 mmol) was dissolved in 100 mM Na2HPO4 aqueous solution (150 mL), and acetonitrile (100 mL) was added, and the pH was adjusted to pH 10-12.5 with 1 N NaOH. Tert-butyl eicosanedioyl-γGlu (2xOEG-OSu) -OtBu (1.59 g, 1.63 mmol) was dissolved in acetonitrile (50 mL) and slowly added to the [Gly8, Arg34]GLP-1-(7-37) peptide solution. The pH was maintained at 10-12.5. After 120 minutes, the reaction mixture was added to water (150 mL) and the pH was adjusted to 5.0 with a 1N HCl aqueous solution. The precipitate was separated by centrifugation and freeze-dried. The crude product was added to a mixed solution of trifluoroacetic acid (60 mL) and dichloromethane (60 ml) and stirred at room temperature for 30 minutes. The mixture was concentrated to about 30 ml, poured into ice-cold n-heptane (300 mL), the precipitated product was separated by filtration, and washed twice with n-heptane. After vacuum drying, the product was purified by ion exchange chromatography (Ressource Q, 0.25%-1.25% ammonium acetate gradient in 42.5% ethanol, pH 7.5), reverse phase chromatography (acetonitrile, water, TFA), the purified fractions were combined, the pH was adjusted to 5.2 with 1N HCl, the precipitate was separated, and freeze-dried to obtain the title compound.

[0140] LC-MS (electrospray): m / z = 1035.8 [M + 4H] 4+

[0141] 2. Preparation of the intermediate tert-butyl docosanoyl-γGlu-(2xOEG-OSu)-OtBu

[0142] 2.1 tert-Butyl docosanoyl-OSu

[0143] Under nitrogen protection, mono-tert-butyl docosane dioic acid ester (20 g, 50.17 mmol) and NHS (5.77 g, 50.17 mmol) were mixed in dichloromethane (400 mL), triethylamine (13.95 mL) was added, and the resulting turbid mixture was stirred at room temperature, and then DCC (11.39 g, 55.19 mmol) was added, and it was further stirred overnight. Filter, concentrate the obtained filtrate to almost dryness, mix the residue with cold water and ethyl acetate, stir for 20 minutes, separate, add saturated brine to wash the upper organic phase, dry the upper organic phase with anhydrous sodium sulfate after separation, and concentrate the filtrate under reduced pressure to almost dryness after filtration, and vacuum dry overnight to obtain 24.12 g (yield 97%) of tert-butyl eicosane dioyl-OSu.

[0144] 2.2 tert-Butyl docosanoyl-γGlu-OtBu

[0145] Tert-butyl docosanoyl-OSu (24.12 g, 48.66 mmol) was dissolved in dichloromethane (250 mL) and stirred, and H-Glu-OtBu (10.88 g, 53.53 mmol), triethylamine (12.49 mL), and water (25 mL) were added in sequence, and heated to obtain a clear solution, which was stirred at room temperature for 4 hours. Then 10% citric acid aqueous solution (200 mL) was added, and the liquid was separated. The lower organic phase was washed with saturated brine. After separation, the lower organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to almost dryness under reduced pressure, and vacuum dried overnight. 27.27 g (yield 96%) of tert-butyl docosanoyl-γGlu-OtBu was obtained.

[0146] 2.3 tert-Butyl docosanoyl-γGlu(OSu)-OtBu.

[0147] Under nitrogen protection, tert-butyl docosane dioyl-γGlu-OtBu (27.27 g, 46.71 mmol) was dissolved in dichloromethane (300 mL), triethylamine (11.99 mL) was added and stirred for 10 minutes, then NHS (5.38 g, 50.17 mmol) was added, followed by DCC (10.60 g, 51.38 mmol). The mixture was stirred overnight at room temperature. Filtered, the obtained filtrate was concentrated to almost dryness, the residue was mixed with cold water and ethyl acetate, stirred for 20 minutes, separated, the upper organic phase was washed with saturated brine, the upper organic phase was dried over anhydrous sodium sulfate after separation, the filtrate was concentrated to almost dryness under reduced pressure after filtration, methyl tert-butyl ether was added, stirred for 30 minutes, filtered, and the filter cake was vacuum dried overnight to obtain 25.76 g (yield 81%) of tert-butyl docosane dioyl-γGlu-(OSu)-OtBu.

[0148] 2.4 tert-Butyl docosanoyl-γGlu-(2xOEG-OH)-OtBu

[0149] Tert-butyl docosanoyl-γGlu-(OSu)-OtBu (25.76 g, 37.83 mmol) was dissolved in dichloromethane (250 mL) and stirred. 2xAEEA (11.66 g, 37.83 mmol), triethylamine (9.71 mL), and water (25 mL) were added in sequence, and the mixture was heated to obtain a clear solution. The solution was stirred at room temperature for 4 hours. Then, 10% citric acid aqueous solution (200 mL) was added to separate the liquids. The lower organic phase was washed with saturated brine. After separation, the lower organic phase was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to almost dryness and vacuum dried overnight. 30.75 g (yield 93%) of tert-butyl docosanoyl-γGlu-(2xOEG-OH)-OtBu was obtained.

[0150] 2.5 tert-Butyldocosanedioyl-γGlu-(2xOEG-OSu)-OtBu.

[0151] Under nitrogen protection, tert-butyl docosanoyl-γGlu-(2xOEG-OH)-OtBu (30.75 g, 35.18 mmol) was dissolved in dichloromethane (300 mL), triethylamine (9.03 mL) was added and stirred for 10 minutes, and NHS (4.05 g, 35.18 mmol) was added, followed by DCC (7.98 g, 38.70 mmol). The mixture was stirred overnight at room temperature. Filtered, the filtrate was concentrated to almost dryness, the residue was mixed with cold water and ethyl acetate, stirred for 20 minutes, separated, the upper organic phase was washed with saturated brine, the upper organic phase was dried with anhydrous sodium sulfate after separation, filtered, the filtrate was concentrated to almost dryness under reduced pressure, and vacuum dried overnight to obtain 31.09 g (yield 91%) of tert-butyl docosanoyl-γGlu-(2xOEG-OSu)-OtBu.

[0152] Example 2 Preparation of PCSK9 Inhibitors

[0153] The present invention exemplarily uses AK102 (Ebronucimab) as a PCSK9 inhibitor to verify the technical effect of the present invention, the heavy chain amino acid sequence of AK102 is shown in SEQ ID NO: 3, and the light chain amino acid sequence is shown in SEQ ID NO: 4. It is prepared by conventional recombinant antibody preparation methods in the art.

[0154] SEQ ID NO: 13

[0155] EVQLVESGGGLVQPGRSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSGISSSSSYISYADSVQGRFTISRDNGKNSL

[0156] YLQMNSLRAEDTALYFCAREYDFWSAYYDAFDVWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF

[0157] PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC

[0158] PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT

[0159] VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG

[0160] QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0161] SEQ ID NO:14

[0162] QSELTQPRSVSGSPGQSVTISCTGTSRNIGGGNDVHWYQQHPGKAPKLLISGVIERSSGVPDRFSGSKSGNTASLTISGLQ

[0163] AEDEADYYCQSFDGSLSGSVFGTGTDVTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSP

[0164] VKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0165] Formulation of AK102 monoclonal antibody preparation

[0166] 1.1 Formulation of AK102 monoclonal antibody preparation

[0167] Table 2

[0168]

[0169] 1.2 AK102 preparation method

[0170] 1.2.1 Preparation of mother solution: prepare 50mM anhydrous disodium hydrogen phosphate and 50mM disodium hydrogen phosphate dodecahydrate respectively;

[0171] 2.2.2 Preparation of buffer: Use a small amount of 50mM anhydrous disodium hydrogen phosphate to adjust the pH of 50mM disodium hydrogen phosphate dodecahydrate to 8.50±0.2, mix thoroughly, and filter and sterilize using a 0.22um disposable filter;

[0172] 1.2.3 Preparation of 200mg / ml sorbitol (5X stock solution): accurately weigh 2g sorbitol, dilute to 10ml with pure water, vortex mix, and filter sterilize using a 0.22um disposable filter;

[0173] 1.2.4 Preparation of 20 mg / ml Tween 80 (200X stock solution): Take 20 ul of Tween 80 stock solution and dissolve it in 980 ul of sterile water. Vortex to mix well and filter with a 0.22 um disposable filter to sterilize.

[0174] 1.2.5 Activate the ultrafiltration tube: add 0.5 ml of sterile water into a 0.5 ml-3 kd ultrafiltration tube, at 4 °C, 13000 rpm, for 10 min to activate the ultrafiltration tube;

[0175] 1.2.6 AK102 protein concentration: according to the experimental requirements, take an appropriate amount of protein solution and concentrate it. Centrifuge each time (4°C, 13000rpm, 25min). When all the proteins are concentrated to a volume of 100ul, the replacement can begin.

[0176] 1.2.7 Follow the method in step 1.2.6, using buffer to replace protein, 5-6 times;

[0177] 1.2.8 Concentration after protein replacement: The fully replaced protein needs to be concentrated to 120 mg / ml;

[0178] 1.2.9 Preparation of AK102 monoclonal antibody preparation (taking 1 ml preparation as an example):

[0179] Table 3

[0180]

[0181] After fully mixing, use a 0.22um disposable filter to filter and sterilize for later use;

[0182] Example 3

[0183] Premix preparation

[0184] 1.3.1 Preparation of premixed preparation (AK102 + compound 1 dry powder): Take 1 ml preparation as an example:

[0185] 12 mg of the compound 1 dry powder prepared in Example 1 was accurately weighed into a 1.5 ml centrifuge tube, and centrifuged instantly to concentrate the dry powder at the bottom of the centrifuge tube. 1 ml of the AK102 monoclonal antibody preparation prepared in Example 2 was added to the centrifuge tube, and gently pipetted and mixed to fully dissolve the compound 1 dry powder to obtain AK102-powder solution (AK102 + compound 1 dry powder).

[0186] Note: (Final preparation component concentration: 95mg / ml AK102 + 12mg / ml compound 1 dry powder)

[0187] 1.3.2 Preparation of liquid-liquid mixed preparation (AK102 + compound 1 liquid): 1 ml preparation as an example

[0188] According to the following formula of Compound 1 liquid preparation, Compound 1 liquid preparation was prepared and sterilized by filtration using a 0.22 um disposable filter;

[0189] Table 4

[0190]

[0191]

[0192] 500ul of the AK102 monoclonal antibody preparation prepared in Example 2 + 500ul of the liquid preparation of compound 1 were mixed evenly in a ratio of 1:1 and set aside for use. AK102-liquid (AK102 + compound 1 liquid) was obtained. Note: (The final concentration of the preparation components is: 47.5mg / ml AK102 + 6mg / ml compound 1).

[0193] Example 4

[0194] AK102-SEC Detection Method

[0195] 1 Sample processing: Dilute the sample to 2 mg / ml with PBS, mix well by pipetting, and filter with a 0.22 um syringe filter for sterilization before testing.

[0196] 2SEC detection method:

[0197] Table 5

[0198]

[0199] 3. Data Collection:

[0200] Data collection starts at 10 minutes and continues until the inverted peak appears. The main peak time of the sample is generally between 15 minutes and 17 minutes.

[0201] 4 Experimental data (day 0)

[0202] Table 6

[0203]

[0204]

[0205] Experimental data showed that the stability of the two forms of premixed preparations was not significantly reduced compared with the AK102 monoclonal antibody preparation, and the stability of AK102 in the powder and liquid form was even better than that of the single AK102 preparation.

[0206] Example 5 Antigen-antibody affinity detection

[0207] 1.1 Experiment name: Antigen-antibody affinity determination

[0208] 1.2 Experimental procedures

[0209] 1.2.11xHBS-P+buffer preparation: dilute 10xHBS-P+buffer to 1x, using

[0210] 0.22μm500mL filter for sterilization and ultrasonic exhaust;

[0211] 1.2.2 Preparation of Regeneration Solution: 10mM Glycine, adjust pH to 3.0 with HCL, and filter sterilize with a 0.22μm filter;

[0212] 1.2.3 Ultrapure water treatment: Use 0.22μm500mL filter to filter and sterilize, then ultrasonically exhaust;

[0213] 1.2.4 Antibody sample preparation: dilute the antibody to 0.5-1ug / ml using 1xHBS-P+buffer (to ensure that the chip captures 100RU of the antibody);

[0214] 1.2.5 Antigen sample preparation: PCSK9 was diluted into 8 concentration gradients using 1xHBS-P+buffer, i.e., 0 nM, 5.3 nM, 7.9 nM, 11.8 nM, 17.7 nM, 26.7 nM, 40 nM, and 60 nM;

[0215] 1.2.6 Method selection Multi-cycle kinetics / affinity using capture;

[0216] 1.3 Experimental Results

[0217] The KD values ​​of the three premixed preparation samples and the dynamic fitting curve results are shown below. The fitting curve diagram is attached. Figure 1 , 2 , 3; The results showed that premixing of monoclonal antibody AK102 with GZR18 powder or liquid did not affect the affinity of AK102 with antigen PCSK9.

[0218] Table 7

[0219]

[0220]

[0221] Example 6 Cell activity detection

[0222] 1. Cell recovery: Take out HepG2 cells from the liquid nitrogen tank and thaw them quickly in a 37°C water bath. Add 9 mL of complete culture medium (DMEM: FBS = 9:1) and mix well. Centrifuge at 200g for 5 min, remove the culture medium, resuspend in fresh complete culture medium, place in a T25 culture flask, and culture in a CO2 incubator.

[0223] 2. Cell culture: When the cell confluence reaches more than 90%, it can be passaged. After removing the culture medium in the cells, add 2mL PBS to clean the culture medium on the cell surface. After aspirating the PBS, add 1mL trypsin and place the cells in an incubator to digest the cells. After the digestion is complete, add 2mL complete culture medium to stop. Pipette and pipette evenly. Passage at a ratio of 1:4, that is, aspirate 750uL of cell suspension into fresh culture medium and place it in an incubator to continue culturing.

[0224] 3. After the P3 generation, the cells can be plated: After the cells are digested according to the cell passaging method, the cell number and viability are counted by a cell counter, and 50,000 cells / well / 100μL are plated. No cells are added to the side wells, but 100μL PBS is used instead. The cells are placed in the incubator for 24 hours. After 24 hours, prepare a DMEM starvation medium with 0.3% BSA, that is, add 1.5mL of 10% BSA to 50mL. Each well is starved and cultured overnight with 100μL of starvation medium.

[0225] 4. System preparation and sample addition

[0226] Take the premixed preparation (AK102 + compound 1 dry powder) and liquid-liquid mixed preparation (AK102 + compound 1 liquid) prepared in Example 3 as preparation samples, draw the starvation medium of the 96-well culture plate, dilute the preparation sample with the starvation medium to 22 μg / ml as the starting concentration, 1:1.5 gradient dilution, take 50 μL and add it to the cell culture plate. Prepare 2.5 μg / mL PCSK9 protein dilution and add it to the corresponding wells (the control wells without PCSK9 are filled with starvation medium). Starve and culture in the incubator for 18-20 hours.

[0227] After 18-20 hours, prepare 2200 μL of 10 μg / mL LDL diluent, that is, take 2.2 mL of starvation culture medium and add 22 μL of BODIPYLDL fluorescent protein, 50 μL per well. Incubate in the incubator for 6 hours. After 6 hours, remove the culture plate, remove all the supernatant, add 200 μL / well of 0.3% BSA DPBS, rinse twice, remove, add 100 μL DPBS and read at 515-540 nm.

[0228] Data processing: Antibody inhibition efficiency = (sample reading - minimum intake average) / (maximum intake average - minimum intake average) * 100%

[0229] Analysis: Use GrapHPad software to fit the data and obtain the fitting curve Figure 4 .

[0230] 4.5.3.10 Cell activity test results

[0231] The plate wells were read at 515-540 nm and the curve fitting results are as follows.

[0232] Table 8:

[0233]

[0234] It can be seen from the results in Table 8 that premixing of monoclonal antibody AK102 with compound 1 powder or liquid does not affect the in vitro cell activity of AK102.

[0235] The present invention has been described through the above embodiments, but it should be understood that the above embodiments are only for the purpose of example and description, and are not intended to limit the present invention to the scope of the described embodiments. In addition, it can be understood by those skilled in the art that the present invention is not limited to the above embodiments, and more variations and modifications can be made according to the teachings of the present invention, and these variations and modifications all fall within the scope of the protection claimed by the present invention. The protection scope of the present invention is defined by the attached claims and their equivalents.

[0236]

[0237]

Claims

1. A drug combination comprising: a) a protein drug for alleviating or treating cholesterol-related symptoms in an amount of 30 mg / ml-250 mg / ml, preferably, the protein drug is selected from PCSK9 inhibitors, ANGPTL3 inhibitors, ANGPTL4 / 8 inhibitors, Apoc3 inhibitors and apolipoprotein (a) inhibitors, and more preferably, the protein drug is a PCSK9 inhibitor; preferably, the protein drug for alleviating or treating cholesterol-related symptoms in an amount of 50 mg / ml-200 mg / ml, and more preferably, 60 mg / ml-180 mg / ml. g / ml, more preferably 70mg / ml-160mg / ml, further preferably 70mg / ml-75mg / ml, 80mg / ml, 85mg / ml, 90mg / ml, 95mg / ml, 100mg / ml, 105mg / ml, 110mg / ml, 115mg / ml, 120mg / ml, 125mg / ml, 130mg / ml, 135mg / ml, 140mg / ml, 145mg / ml, 150mg / ml, 155mg / ml or 160mg / ml; or The content of the protein drug for alleviating or treating cholesterol-related symptoms is 30mg / ml-120mg / ml, more preferably 40mg / ml-110mg / ml, more preferably 45mg / ml-100mg / ml, and more preferably 45mg / ml, 47.5mg / ml, 50mg / ml, 52.5mg / ml, 55mg / ml, 57.5mg / ml, 60mg / ml, 62.5mg / ml, 65mg / ml, 67.5mg / ml, 70mg / ml, 72.5mg / ml, 75mg / ml, 77.5mg / ml, 80mg / ml, 82.5mg / ml, 85mg / ml, 87.5mg / ml, 90mg / ml, 92.5mg / ml, 95mg / ml, 97.5mg / ml or 100mg / ml; preferably, the The PCSK9 inhibitor is an antibody or an antigen-binding fragment thereof that specifically binds to PCSK9; preferably, the PCSK9 inhibitor is selected from at least one of alirocumab, evolocumab, lodelcizumab, ralpancizumab, bococizumab, ebronucimab, LY3015014, LIB-003, SHR-1209, JS-002, SAL-003 and ATH-06; preferably, the PCSK9 inhibitor is alirocumab, evolocumab, lodelcizumab, ralpancizumab, bococizumab, ebronucimab, LY3015014, LIB-003, SHR-1209, JS-002, SAL-003 or ATH-06; and b) A polypeptide drug for lowering blood sugar or reducing weight with a content of 2mg / ml-60mg / ml. Preferably, the polypeptide drug is selected from GLP-1 receptor agonists, GIP receptor agonists, GCCR agonists, FGF21 receptor agonists, and agonists targeting any two or three targets of GLP-1, GIP, GCCR and FGF21. Further preferably, the polypeptide drug is a GLP-1 receptor agonist, a GLP-1R / GIPR dual agonist, a GLP-1R / GCGR dual agonist, a GLP-1R / FGF21R ​​dual agonist or a GLP-1R / GIPR / GCCR triple agonist; preferably, the content of the polypeptide drug for lowering blood sugar or reducing weight is 2mg / ml-50mg / ml, more preferably 3mg / ml-40mg / ml, further preferably 3mg / ml, 3.5mg / ml, 4mg / ml, 4.5mg / ml, 5mg / ml, 5.5mg / ml, 6mg / ml, 6.5mg / ml, 7mg / ml, 7.5mg / ml, 8mg / ml, 8.5mg / ml, 9mg / ml, 9.5mg / ml, 10mg / ml, 10.5mg / ml, 11mg / ml, 11.5mg / ml, 12mg / ml, 12.5mg / ml, 13mg / ml, 13.5mg / ml, 14mg / ml, 14.5mg / ml, 15mg / ml, 15.5 mg / ml, 16mg / ml, 16.5mg / ml, 17mg / ml, 17.5mg / ml, 18mg / ml, 18.5mg / ml, 19mg / ml, 19.5mg / ml, 20mg / ml, 20.5mg / ml, 21mg / ml, 21.5mg / ml, 22mg / ml, 2 2.5mg / ml, 23mg / ml, 23.5mg / ml, 24mg / ml, 24.5mg / ml, 25mg / ml, 25.5mg / ml, 26mg / ml, 26.5mg / ml, 27mg / ml, 27.5mg / ml, 28mg / ml, 28.5mg / ml, 29mg / m l, 29.5mg / ml, 30mg / ml, 31.5mg / ml, 32mg / ml, 32.5mg / ml, 33mg / ml, 33.5mg / ml, 34mg / ml, 34.5mg / ml, 35mg / ml, 35.5mg / ml, 36mg / ml, 36.5mg / ml, 37m g / ml, 37.5mg / ml, 38mg / ml, 38.5mg / ml, 39mg / ml, 39.5mg / ml, 40mg / ml, 40mg / ml, 41.5mg / ml, 42mg / ml, 42.5mg / ml, 43mg / ml, 43.5mg / ml, 44mg / ml, 44.5mg / ml, 45mg / ml, 45.5mg / ml, 46mg / ml, 46.5mg / ml, 47mg / ml, 47.5mg / ml, 48mg / ml, 48.5mg / ml, 49mg / ml, 4 9.5mg / ml, 50mg / ml, 50mg / ml, 51.5mg / ml, 52mg / ml, 52.5mg / ml, 53mg / ml, 53.5mg / ml, 54mg / ml, 54.5mg / ml, 55mg / ml, 55.5mg / ml, 56mg / ml, 56.5mg / ml, 57mg / ml, 57.5mg / ml, 58mg / ml, 58.5mg / ml, 59mg / ml, 59.5mg / ml, or 60mg / ml; preferably, the hypoglycemic or weight-reducing polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tirzepatide, exenatide, liraglutide, lixisenatide, N-ε. 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai and glutazumab.

2. The combination according to claim 1, comprising a) The content is 60mg / ml-180mg / ml, more preferably 70mg / ml-160mg / ml, and further preferably 70mg / ml, 75mg / ml, 80mg / ml, 85mg / ml, 90mg / ml, 95mg / ml, 100mg / ml, 105mg / ml, 110mg / ml, 115mg / ml, 120mg / ml, 125mg / ml, 130mg / ml, 135mg / ml, 140 100 mg / ml, 145 mg / ml, 150 mg / ml, 155 mg / ml or 160 mg / ml of a PCSK9 inhibitor selected from at least one of alirocumab, evolocumab, lodelcizumab, ralpancizumab, bococizumab, inusituzumab, LY3015014, LIB-003, SHR-1209, JS-002, SAL-003 and ATH-06; Or the content is 30mg / ml-120mg / ml, more preferably 40mg / ml-110mg / ml, more preferably 45mg / ml-100mg / ml, and further preferably 45mg / ml, 47.5mg / ml, 50mg / ml, 52.5mg / ml, 55mg / ml, 57.5mg / ml, 60mg / ml, 62.5mg / ml, 65mg / ml, 67.5mg / ml, 70mg / ml, 72.5mg / ml, 75mg / ml, 77.5mg / ml, 80mg / ml , 82.5 mg / ml, 85 mg / ml, 87.5 mg / ml, 90 mg / ml, 92.5 mg / ml, 95 mg / ml, 97.5 mg / ml or 100 mg / ml of a PCSK9 inhibitor, wherein the PCSK9 inhibitor is selected from at least one of Alisikumab, Elotumab, Lodelcizumab, Ralpancizumab, Bococizumab, Inusinumab, LY3015014, LIB-003, SHR-1209, JS-002, SAL-003 and ATH-06. b) The content is 2 mg / ml - 50 mg / ml, more preferably 3 mg / ml - 40 mg / ml, and even more preferably 3 mg / ml, 3.5 mg / ml, 4 mg / ml, 4.5 mg / ml, 5 mg / ml, 5.5 mg / ml, 6 mg / ml, 6.5 mg / ml, 7 mg / ml, 7.5 mg / ml, 8 mg / ml, 8.5 mg / ml, 9 mg / ml, 9.5 mg / ml, 10 mg / ml, 10.5 mg / ml, 11 mg / ml, 11.5 mg / ml, 12 mg / ml, 12.5 mg / ml, 13 mg / ml, 13.5 mg / ml, 14 mg / ml, 14.5 mg / ml, 15 mg / ml, 15.5 mg / ml, 16 mg / ml, 16.5 mg / ml, 17 mg / ml, 17.5 mg / ml, 18 mg / ml, 18.5 mg / ml, 19 mg / ml, 19.5 mg / ml, 20 mg / ml, 20.5 mg / ml, 21 mg / ml, 21.5 mg / ml, 22 mg / ml, 22.5 mg / ml, 23 mg / ml, 23.5 mg / ml, 24 mg / ml, 24.5 mg / ml, 25 mg / ml, 25.5 mg / ml, 26 mg / ml, 26.5 mg / ml, 27 mg / ml, 27.5 mg / ml, 28 mg / ml, 28.5 mg / ml, 29 mg / ml, 29.5 mg / ml, 30 mg / ml, 31.5 mg / ml, 32 mg / ml, 32.5 mg / ml, 33 mg / ml, 33.5 mg / ml, 34 mg / ml, 34.5 mg / ml, 35 mg / ml, 35.5 mg / ml, 36 mg / ml, 36.5 mg / ml, 37 mg / ml, 37.5 mg / ml, 38 mg / ml, 38.5 mg / ml, 39 mg / ml, 39.5 mg / ml, 40 mg / ml, 40 mg / ml, 41.5 mg / ml, 42 mg / ml, 42.5 mg / ml, 43 mg / ml, 43.5 mg / ml, 44 mg / ml, 44.5 mg / ml, 45 mg / ml, 45.5 mg / ml, 46 mg / ml, 46.5 mg / ml, 47 mg / ml, 47.5 mg / ml, 48 mg / ml, 48.5 mg / ml, 49 mg / ml, 49.5 mg / ml, 50 mg / ml, 50 mg / ml, 51.5 mg / ml, 52 mg / ml, 52.5 mg / ml, 53 mg / ml, 53.5 mg / ml, 54 mg / ml, 54.5 mg / ml, 55 mg / ml, 55.5 mg / ml, 56 mg / ml, 56.5 mg / ml, 57 mg / ml, 57.5 mg / ml, 58 mg / ml, 58.5mg / ml, 59mg / ml, 59.5mg / ml, or 60mg / ml of the hypoglycemic or weight-reducing polypeptide drug, wherein the hypoglycemic or weight-reducing polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε. 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai and glutazumab.

3. A drug combination comprising: a) Ebronucimab. Preferably, the content of Ebronucimab is 60 mg / ml-180 mg / ml, more preferably 70 mg / ml-160 mg / ml, and further preferably 70 mg / ml, 75 mg / ml, 80 mg / ml, 85 mg / ml, 90 mg / ml, 95 mg / ml, 100 mg / ml, 105 mg / ml, 110 mg / ml, 115 mg / ml, 120 mg / ml, 125 mg / ml, 130 mg / ml, 135 mg / ml, 140 mg / ml, 145 mg / ml, 150 mg / ml, 155 mg / ml or 160 mg / ml; or the content is 30mg / ml-120mg / ml, more preferably 40mg / ml-110mg / ml, further preferably 45mg / ml-100mg / ml, further preferably 45mg / ml, 47.5mg / ml, 50mg / ml, 52.5mg / ml, 55mg / ml, 57.5mg / ml, 60mg / ml, 62.5mg / ml, 65mg / ml, 67.5mg / ml, 70mg / ml, 72.5mg / ml, 75mg / ml, 77.5mg / ml, 80mg / ml, 82.5mg / ml, 85mg / ml, 87.5mg / ml, 90mg / ml, 92.5mg / ml, 95mg / ml, 97.5mg / ml or 100mg / ml; b) A polypeptide drug for lowering blood sugar or reducing weight, preferably, the polypeptide drug is selected from GLP-1 receptor agonists, GIP receptor agonists, GCCR agonists, FGF21 receptor agonists and agonists targeting any two or three targets of GLP-1, GIP, GCCR and FGF21 at the same time, and further preferably, the polypeptide drug is a GLP-1 receptor agonist, a GLP-1R / GIPR dual agonist, a GLP-1R / GCGR dual agonist, a GLP-1R / FGF21R ​​dual agonist or a GLP-1R / GIPR / GCCR triple agonist; preferably, the polypeptide drug for lowering blood sugar or reducing weight is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai and glutazumab; preferably, the content of the hypoglycemic polypeptide drug is 2 mg / ml-50 mg / ml, more preferably 3 mg / ml -40mg / ml, more preferably 3mg / ml, 3.5mg / ml, 4mg / ml, 4.5mg / ml, 5mg / ml, 5.5mg / ml, 6mg / ml, 6.5mg / ml, 7mg / ml, 7.5mg / ml, 8mg / ml, 8.5mg / ml, 9mg / ml, 9.5m g / ml, 10mg / ml, 10.5mg / ml, 11mg / ml, 11.5mg / ml, 12mg / ml, 12.5mg / ml, 13mg / ml, 13.5mg / ml, 14mg / ml, 14.5mg / ml, 15mg / ml, 15.5mg / ml, 16mg / ml, 16. 5mg / ml, 17mg / ml, 17.5mg / ml, 18mg / ml, 18.5mg / ml, 19mg / ml, 19.5mg / ml, 20mg / ml, 20.5mg / ml, 21mg / ml, 21.5mg / ml, 22mg / ml, 22.5mg / ml, 23mg / ml, 23.5mg / ml, 24mg / ml, 24.5mg / ml, 25mg / ml, 25.5mg / ml, 26mg / ml, 26.5mg / ml, 27mg / ml, 27.5mg / ml, 28mg / ml, 28.5mg / ml, 29mg / ml, 29.5mg / ml, 30mg / m l, 31.5mg / ml, 32mg / ml, 32.5mg / ml, 33mg / ml, 33.5mg / ml, 34mg / ml, 34.5mg / ml, 35mg / ml, 35.5mg / ml, 36mg / ml, 36.5mg / ml, 37mg / ml, 37.5mg / ml, 38m g / ml, 38.5mg / ml, 39mg / ml, 39.5mg / ml, 40mg / ml, 40mg / ml, 41.5mg / ml, 42mg / ml, 42.5mg / ml, 43mg / ml, 43.5mg / ml, 44mg / ml, 44.5mg / ml, 45mg / ml, 45.5 mg / ml, 46 mg / ml, 46.5 mg / ml, 47 mg / ml, 47.5 mg / ml, 48 mg / ml, 48.5 mg / ml, 49 mg / ml, 49.5 mg / ml, 50 mg / ml, 50 mg / ml, 51.5 mg / ml, 52 mg / ml, 52.5 mg / ml, 53 mg / ml, 53.5 mg / ml, 54 mg / ml, 54.5 mg / ml, 55 mg / ml, 55.5 mg / ml, 56 mg / ml, 56.5 mg / ml, 57 mg / ml, 57.5 mg / ml, 58 mg / ml, 58.5 mg / ml, 59 mg / ml, 59.5 mg / ml, or 60 mg / ml.

4. The combination according to claim 3, comprising a) 60mg / ml-180mg / ml, more preferably 70mg / ml-160mg / ml, more preferably 70mg / ml, 75mg / ml, 80mg / ml, 85mg / ml, 90mg / ml, 95mg / ml, 100mg / ml, 105mg / ml, 110mg / ml, 115mg / ml, 120mg / ml, 125mg / ml, 130mg / ml, 135mg / ml, 140mg / ml, 145mg / ml, 150mg / ml, 155mg / ml or 160mg / ml of inuncimab; or 30mg / ml-120mg / ml, more preferably 40mg / ml -110 mg / ml, more preferably 45 mg / ml-100 mg / ml, further preferably 45 mg / ml, 47.5 mg / ml, 50 mg / ml, 52.5 mg / ml, 55 mg / ml, 57.5 mg / ml, 60 mg / ml, 62.5 mg / ml, 65 mg / ml, 67.5 mg / ml, 70 mg / ml, 72.5 mg / ml, 75 mg / ml, 77.5 mg / ml, 80 mg / ml, 82.5 mg / ml, 85 mg / ml, 87.5 mg / ml, 90 mg / ml, 92.5 mg / ml, 95 mg / ml, 97.5 mg / ml or 100 mg / ml of inunusimab; b) 2 mg / ml - 50 mg / ml, further preferably 3 mg / ml - 40 mg / ml, still further preferably 3 mg / ml, 3.5 mg / ml, 4 mg / ml, 4.5 mg / ml, 5 mg / ml, 5.5 mg / ml, 6 mg / ml, 6.5 mg / ml, 7 mg / ml, 7.5 mg / ml, 8 mg / ml, 8.5 mg / ml, 9 mg / ml, 9.5 mg / ml, 10 mg / ml, 10.5 mg / ml, 11 mg / ml, 11.5 mg / ml, 12 mg / ml, 12.5 mg / ml, 13 mg / ml, 13.5 mg / ml, 14 mg / ml, 14.5 mg / ml, 15 mg / ml, 15.5 mg / ml, 16 mg / ml, 16.5 mg / ml, 17 mg / ml, 17.5 mg / ml, 18 mg / ml, 18.5 mg / ml, 19 mg / ml, 19.5 mg / ml, 20 mg / ml, 20.5 mg / ml, 21 mg / ml, 21.5 mg / ml, 22 mg / ml, 22.5 mg / ml, 23 mg / ml, 23.5 mg / ml, 24 mg / ml, 24.5 mg / ml, 25 mg / ml, 25.5 mg / ml, 26 mg / ml, 26.5 mg / ml, 27 mg / ml, 27.5 mg / ml, 28 mg / ml, 28.5 mg / ml, 29 mg / ml, 29.5 mg / ml, 30 mg / ml, 31.5 mg / ml, 32 mg / ml, 32.5 mg / ml, 33 mg / ml, 33.5 mg / ml, 34 mg / ml, 34.5 mg / ml, 35 mg / ml, 35.5 mg / ml, 36 mg / ml, 36.5 mg / ml, 37 mg / ml, 37.5 mg / ml, 38 mg / ml, 38.5 mg / ml, 39 mg / ml, 39.5 mg / ml, 40 mg / ml, 40 mg / ml, 41.5 mg / ml, 42 mg / ml, 42.5 mg / ml, 43 mg / ml, 43.5 mg / ml, 44 mg / ml, 44.5 mg / ml, 45 mg / ml, 45.5 mg / ml, 46 mg / ml, 46.5 mg / ml, 47 mg / ml, 47.5 mg / ml, 48 mg / ml, 48.5 mg / ml, 49 mg / ml, 49.5 mg / ml, 50 mg / ml, 50 mg / ml, 51.5 mg / ml, 52 mg / ml, 52.5 mg / ml, 53 mg / ml, 53.5 mg / ml, 54 mg / ml, 54.5 mg / ml, 55 mg / ml, 55.5 mg / ml, 56 mg / ml, 56.5 mg / ml, 57 mg / ml, 57.5 mg / ml, 58 mg / ml, 58.5mg / ml, 59mg / ml, 59.5mg / ml, or 60mg / ml of a polypeptide drug for lowering blood sugar or reducing weight, wherein the polypeptide drug for lowering blood sugar or reducing weight is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε. 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8, Arg34] GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai, and glutazumab.

5. A combination as claimed in any one of claims 1 to 4, wherein: The PCSK9 inhibitor and the polypeptide drug for lowering blood sugar or reducing weight are administered simultaneously or separately.

6. A combination as claimed in any one of claims 1 to 5, wherein: The combination is in the form of a pharmaceutical composition or a kit.

7. A pharmaceutical composition comprising the drug according to any one of claims 1 to 5.

8. The pharmaceutical composition according to claim 1, wherein The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients; preferably, the pharmaceutically acceptable excipients are selected from at least one of a buffer, a stabilizer, a surfactant, and an isotonic agent; Preferably, The buffer is selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, histidine, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer is selected from at least one of citric acid, sodium acetate and disodium hydrogen phosphate; preferably the buffer is a combination of citric acid and disodium hydrogen phosphate; preferably, the buffer is a combination of sodium dihydrogen phosphate and disodium hydrogen phosphate; preferably, the content of the buffer in the combination is 0.05mM-about 100mM, preferably about 0.05mM-about 85mM, preferably about 0.05mM-about 82mM, preferably about 0.05mM-about 40mM, preferably about 0.1mM-about 30mM, preferably about 1mM-about 20mM, preferably about 5mM-about 15mM, preferably about 5mM-about 10mM, preferably about 30-50mM, preferably about 35-45mM, preferably about 40mM; and / or The stabilizer is selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably, selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine and proline; preferably selected from at least one of histidine, proline and sucrose; preferably selected from sorbitol; preferably, the content of the stabilizer in the combination is about 0.1% w / v-about 15% w / v, preferably about 0.5% w / v-about 10% w / v, preferably about 2.5% w / v-about 10% w / v, preferably about 3% w / v-about 5% w / v, preferably about 4% w / v; and / or The surfactant is selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188, preferably selected from polysorbate 80. Preferably, the content of the surfactant in the combination is about 0.001% w / v to about 10% w / v, preferably about 0.01% w / v to about 1% w / v; preferably about 0.01% w / v to about 0.05% w / v; preferably about 0.01% w / v; Preferably, the pH of the drug combination is about 4.0-about 8.0, preferably about 4.5-about 7.5, preferably about 4.5-about 7.0, preferably about 4.5-about 6.5, preferably about 5.6-about 6.2, preferably about 4.5-about 5.5, preferably about 5.0-about 5.5; preferably about 7.0-8.0, preferably about 7.3-8.0, preferably about 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.

0.

9. The pharmaceutical composition according to claim 1, wherein The combination of a) and b) in the pharmaceutical combination further comprises one or more pharmaceutically acceptable excipients; preferably, the pharmaceutically acceptable excipient is selected from at least one of a buffer, a stabilizer, a surfactant, and an isotonic agent; Preferably, wherein component a) further comprises A buffer selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, histidine, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer selected from at least one of citric acid, sodium acetate, and disodium hydrogen phosphate; preferably the buffer is a combination of citric acid and disodium hydrogen phosphate; preferably, the buffer is a combination of sodium dihydrogen phosphate and disodium hydrogen phosphate; A stabilizer selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine, and proline; preferably selected from at least one of histidine, proline, and sucrose; preferably selected from sorbitol; A surfactant selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188, preferably selected from polysorbate 80; and / or wherein component b) further comprises A buffer selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, histidine, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer selected from at least one of citric acid, sodium acetate, and disodium hydrogen phosphate; preferably the buffer is a combination of citric acid and disodium hydrogen phosphate; preferably, the buffer is a combination of citric acid and disodium hydrogen phosphate; The isotonic agent is at least one selected from the group consisting of sodium chloride, propylene glycol, and glycerol, preferably sodium chloride.

10. A pharmaceutical composition, comprising: The pharmaceutical combination according to claims 1 to 14; A buffer, wherein the buffer is selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer is selected from at least one of citric acid, sodium acetate, and disodium hydrogen phosphate, preferably the buffer is a combination of citric acid and disodium hydrogen phosphate; preferably the buffer is a combination of sodium dihydrogen phosphate and disodium hydrogen phosphate; A stabilizer, the stabilizer is selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably, selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine, histidine and proline; preferably, the stabilizer is selected from at least one of histidine, proline and sucrose; preferably, sorbitol; and A surfactant, wherein the surfactant is selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188; preferably polysorbate 80 Preferably, The pH of the pharmaceutical composition is about 4.0 to about 8.0, preferably about 4.5 to about 7.5, preferably about 4.5 to about 7.0, preferably about 4.5 to about 6.5, preferably about 4.5 to about 6.2, preferably about 5.6 to about 6.2, preferably about 6.0 to about 6.2, preferably about 4.5 to about 5.5, preferably about 5.0 to about 5.5, preferably about 7.0-8.0, preferably about 7.3-8.0, preferably about 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.

0.

11. A pharmaceutical preparation, which is prepared by the pharmaceutical combination or pharmaceutical composition according to claims 1 to 22; preferably, the pharmaceutical preparation is a lyophilized preparation or a liquid preparation.

12. A kit, comprising: (1) A first therapeutic agent: comprising the polypeptide drug for lowering blood sugar or reducing weight as described in claim 6; Preferably, the hypoglycemic or weight-reducing polypeptide drug is selected from polyethylene glycol loxenatide, dulaglutide, semaglutide, albiglutide, tesiparatide, exenatide, liraglutide, lixisenatide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, cagrilintide, mazdutide, avexitide, ecnoglutide, retatrutide, efpeglenatide, NN-1535, rE-4, PB-119, CJC-1134-PC, supalutai and glutazumab; Preferably, the first therapeutic agent is a lyophilized powder preparation or a liquid preparation. When it is a liquid preparation, it further comprises: A buffer selected from at least one of citric acid, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, histidine, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffers, preferably the buffer selected from at least one of citric acid, sodium acetate, and disodium hydrogen phosphate; preferably the buffer is a combination of citric acid and disodium hydrogen phosphate; preferably, the buffer is a combination of citric acid and disodium hydrogen phosphate; An isotonic agent, selected from at least one of sodium chloride, propylene glycol, and glycerol, preferably sodium chloride; (2) a second therapeutic agent comprising the PCSK9 inhibitor of claim 5; Preferably, the PCSK9 inhibitor is alirocumab, evolocumab or inusumab. Preferably, the second therapeutic agent further comprises: A buffer, wherein the buffer is selected from at least one of citric acid monohydrate, citrate, sodium acetate, sodium dihydrogen phosphate, glutamate, disodium hydrogen phosphate, and tris(hydroxymethylaminomethane) buffer, preferably the buffer is selected from at least one of citric acid monohydrate, sodium acetate, and disodium hydrogen phosphate; preferably, it is a combination of disodium hydrogen phosphate and sodium dihydrogen phosphate; A stabilizer, the stabilizer is selected from at least one of polyhydroxy hydrocarbons, disaccharides, benzyl alcohol, amino acids, polyols, and phenols, preferably, selected from at least one of sucrose, lactose, fructose, maltose, trehalose, sorbitol, mannitol, arginine, lysine, methionine, taurine, histidine and proline; preferably, the stabilizer is selected from at least one of histidine, proline and sucrose; preferably, sorbitol; and A surfactant, wherein the surfactant is selected from at least one of polysorbate 80, polysorbate 20, and poloxamer 188; preferably polysorbate 80.

13. Use of the pharmaceutical combination, composition or kit according to any one of claims 1 to 28 in the preparation of a medicament for treating hyperglycemia, diabetes, obesity, cholesterol-related diseases, hypercholesterolemia, and / or hyperlipidemia.

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