Pharmaceutical compositions of dual GLP-1 and GIP receptor agonists and uses thereof

Abstract

The present invention provides a pharmaceutical composition of a GLP-1 receptor and a GIP receptor dual agonist and its use. Specifically, the pharmaceutical composition comprises a GLP-1 analogue represented by the general formula (I) and a buffer such as a phosphate buffer, and the composition may further comprise an osmotic pressure adjusting agent such as propylene glycol, sodium chloride or mannitol. The pharmaceutical composition has good biological activity and stability. General formula (I): R1-X1-X2-Glu-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -Glu-Phe-X 23 -X 24 -Trp-Leu-X 27 -X 28 -X 29 -X 30 -Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2 (I)

Description

[Technical field]

[0001] The present disclosure relates to the field of pharmaceutical formulations, specifically to pharmaceutical compositions comprising GLP-1 and GIP receptor dual agonists and their pharmaceutical uses. [Background technology]

[0002] At present, many GLP-1 class drugs, such as liraglutide, semaglutide and dulaglutide, are available on the market, mainly used for the treatment of T2DM, of which liraglutide has also been approved for marketing by the FDA as an anti-obesity drug. Under physiological conditions, active GLP-1 consists of 30 amino acids and is secreted from intestinal L cells after a meal by cleaving glucagon proate with PC1 / 3 enzyme. In T2DM patients, the secretion of GLP-1 after a meal is significantly suppressed, but the response of the GLP-1R of the patients to GLP-1 at pharmacological concentrations is not significantly different from that of normal people, further demonstrating the greater therapeutic potential of this target. GLP-1R agonists have significant advantages in therapeutic effects such as glucose reduction and weight loss, but their effects on the central nervous system and stomach cause nausea and vomiting, i.e., dose-dependent gastrointestinal side effects. The therapeutic dose of GLP-1 class drugs is limited, and by continuing to increase the dose, more significant glucose reduction, weight reduction and other therapeutic effects cannot be achieved, so it is necessary to supplement other therapeutic plans to enhance the therapeutic effect or reduce the incidence of side effects of GLP-1 class drugs.

[0003] Glucose-dependent insulinotropic polypeptide (GIP) also belongs to incretin. Active GIP contains 42 amino acids. By cleaving GIP precursor by PC1 / 3 enzyme in enteroendocrine K cells, it can simultaneously exert a comprehensive regulatory effect on the nervous system and endocrine system, and also plays a role in improving metabolism. GLP-1 exerts its effect by directly affecting the central nervous system, pancreas, and stomach, and indirectly acting on the liver, and can also exert a comprehensive metabolic improvement effect by superimposing active ingredients that exert their effect on fat and muscle tissue. Research has revealed that in the body of non-insulin-dependent diabetes patients, the incretin function of GIP polypeptide is greatly reduced, causing the patients to lack or lose the incretin effect. Research has revealed that when blood glucose level is restored to normal, the inhibitory effect of GIP polypeptide generated in these diabetes patients is greatly weakened.

[0004] Therefore, in order to restore the tolerance of non-insulin-dependent diabetes mellitus patients to GIP polypeptides and to obtain a stronger clinical hypoglycemic effect in accordance with the incretin effect of GIP polypeptides, a clinically effective combination of a treatment method for non-insulin-dependent diabetes mellitus using GIP polypeptides and a clinically effective hypoglycemic drug is required.

[0005] PCT / CN2021 / 096568 provides derivatives of GLP-1 analogs with dual agonistic effects on human GLP-1 receptors and human GIP receptors, and agonistic activity on human GIP receptors. Compared with GLP-1 receptor agonists known in the art, they have stronger therapeutic effects on blood glucose reduction and weight loss, have extremely high plasma stability, and have pharmacokinetic properties that allow them to be administered to human subjects by subcutaneous injection once a week. However, chemically modified polypeptide drugs have complex structures and are prone to instability due to degradation, polymerization, or undesirable chemical modification, etc., so that they are suitable for administration, maintain stability during storage and subsequent use, and exert higher therapeutic effects, and therefore it is particularly important to research stable formulations of chemically modified polypeptide drugs. Summary of the Invention

[0006] The present disclosure relates to (a) a GLP-1 analogue represented by general formula (I) or a medicamentous salt thereof; R 1 -X 1 -X 2 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -Glu-Phe-X 23 -X 24 -Trp-Leu-X 27 -X 28 -X 29 -X 30 -Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R 2 (I) (SEQ ID NO: 19) Among them, R 1 is hydrogen (H), an alkyl group, an acetyl group, a formyl group, a benzoyl group, a trifluoroacetyl group, pGlu, or is absent, R 2 Ha-NH 2 , -OH or absent, X 1 , X 2 , X 10 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 23 , X 24 , X 27 , X 28 , X 29 and X 30 are independently selected from any natural amino acid residue, any unnatural amino acid residue, or any peptide fragment consisting of natural and / or unnatural amino acid residues; and The pharmaceutical composition includes at least one selected from (b) a buffering agent, (c) an osmotic pressure adjusting agent, (d) a pH adjusting agent, and (e) a bacteriostatic agent.

[0007] In some embodiments, the pharmaceutical composition comprises a GLP-1 analogue as shown in general formula (I) or a medicamentable salt thereof and a buffering agent.

[0008] In some embodiments, the buffer is one or more selected from an acetate buffer, a histidine buffer, a phosphate buffer, a succinate buffer, and a citrate buffer.

[0009] In some embodiments, the buffer is a phosphate buffer, such as disodium hydrogen phosphate.

[0010] In some embodiments, X 1 is Tyr or His, and X 2 is Aib or D-Ala, and X 10 is Val or Tyr, and X 12 is Ser or Ile, and X 13 is Tyr or Ala, and X 14 is Leu or Nle, and X 15is Asp or Glu, and X 16 is Arg, Glu, Gly, Lys or Aib, and X 17 is Glu, Ile or Gln, and X 18 is Ala, Aib or His, and X 19 is Ala, Aib or Gln, and X 20 is Gln, Glu, Lys, and X 23 is Ile or Val, and X 24 is Ala, Asn or Gln, and X 27 is Val or Leu, and X 28 is Arg or Ala, and X 29 is Gly or Gln, and X 30 are Gly and Lys.

[0011] In some embodiments, among these, X 1 is Tyr or His, and X 2 is Aib or D-Ala, and X 10 is Val or Tyr or Y1, and X 12 is Ser or Ile or Y1, and X 13 is Tyr or Ala or Y1, and X 14 is Leu or Nle or Y1, and X 15 is Asp or Glu, and X 16 is Arg, Glu, Gly, Lys, Aib, or Y1, and X 17 is Glu, Ile, Gln, or Y1, and X 18 is Ala, Aib or His, and X 19 is Ala, Aib or Gln, and X 20 is Gln, Glu, Lys, and X 23 is Ile or Val, and X 24 is Ala, Asn or Gln, and X 27 is Val or Leu, and X 28 is Arg or Ala, and X 29 is Gly or Gln, and X 30is Gly, Lys, and Y1 is a substituted Lys, Orn, Dap, Dab or Cys residue, for example having a modifying group in the side chain of the Lys, Orn, Dap, Dab or Cys residue.

[0012] In some embodiments, Y1 is a Lys, Orn, Dap, Dab, or Cys residue having a substituent at its side chain, the substituent having the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In the structure, a is an integer of 1 to 3 (including 1, 2, and 3), b is 1 or 2, and c is an integer of 10 to 30 (including 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30).

[0013] In some embodiments, among these, X 1 is Tyr, and X 2 is Aib, and X 10 is Tyr, and X 12 is Ile, and X 13 is Tyr, and X 14 is Y1 and X 15 is Asp or Glu, and X 16 is Arg or Lys, and X 17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln, and X 23 is Ile or Val, and X 24 is Asn, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, and Y1 is a group having a side chain of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) cIt is a Lys, Orn, Dap, Dab or Cys residue linked to a substituent having —COOH, wherein a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0014] In some embodiments, among these, X 1 is Tyr, and X 2 is Aib, and X 10 is Tyr, and X 12 is Ile, and X 13 is Tyr, and X 14 is Y1 and X 15 is Asp or Glu, and X 16 is Lys, and X 17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln, and X 23 is Val and X 24 is Asn, and X 27 is Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0015] In some embodiments, X 1 is Tyr, and X 2 is Aib or D-Ala, and X 10 is Y1 and X 12 is Ile, and X 13 is Tyr, and X 14 is Leu or Nle, and X 15 is Glu, and X 16 is Arg or Lys, and X 17 is Ile, and X 18is Ala, and X 19 is Ala, and X 20 is Gln or Lys, and X 23 is Ile or Val, and X 24 is Asn or Gln, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0016] In some embodiments, X 1 is Tyr, and X 2 is Aib or D-Ala, and X 10 is Tyr, and X 12 is Y1 and X 13 is Tyr, and X 14 is Leu or Nle, and X 15 is Glu, and X 16 is Arg or Lys, and X 17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln or Lys, and X 23 is Ile or Val, and X 24 is Asn or Gln, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0017] In some embodiments, X 1 is Tyr, and X 2 is Aib or D-Ala, and X 10 is Tyr, and X 12 is Ile, and X 13 is Y1 and X 14 is Leu or Nle, and X 15 is Glu, and X 16 is Arg or Lys, and X 17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln or Lys, and X 23 is Ile or Val, and X 24 is Asn or Gln, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0018] In some embodiments, X 1 is Tyr, and X 2 is Aib or D-Ala, and X 10 is Tyr, and X 12 is Ile, and X 13 is Tyr, and X 14 is Y1 and X 15 is Glu, and X 16 is Arg or Lys, and X17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln or Lys, and X 23 is Ile or Val, and X 24 is Asn or Gln, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0019] In some embodiments, X 1 is Tyr, and X 2 is Aib, and X 10 is Tyr, and X 12 is Ile, and X 13 is Tyr, and X 14 is Y1 and X 15 is Glu, and X 16 is Arg or Lys, and X 17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln, and X 23 is Ile or Val, and X 24 is Asn, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b-CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0020] In some embodiments, X 1 is Tyr, and X 2 is Aib or D-Ala, and X 10 is Tyr, and X 12 is Ile, and X 13 is Tyr, and X 14 is Leu or Nle, and X 15 is Glu, and X 16 is Y1 and X 17 is Ile, and X 18 is Ala, and X 19 is Ala, and X 20 is Gln or Lys, and X 23 is Ile or Val, and X 24 is Asn or Gln, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0021] In some embodiments, X 1 is Tyr, and X 2 is Aib or D-Ala, and X 10 is Tyr, and X 12 is Ile, and X 13 is Tyr, and X 14 is Leu or Nle, and X 15 is Glu, and X 16 is Arg or Lys, and X17 is Y1 and X 18 is Ala, and X 19 is Ala, and X 20 is Gln or Lys, and X 23 is Ile or Val, and X 24 is Asn or Gln, and X 27 is Ile or Leu, and X 28 is Ala, and X 29 is Gly, and X 30 is Gly, Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0022] In some embodiments, X 10 , X 12 , X 13 , X 14 , X 16 and X 17 are each independently selected from Y1, where Y1 is a Lys, Orn, Dap, Dab or Cys residue having a substituent at the side chain, the substituent being represented by the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c In this structure, a is an integer of 1 to 3, b is 1 or 2, and c is an integer of 10 to 30.

[0023] In some embodiments, a is 2, b is 1 or 2, and c is an integer from 16 to 20 (eg, c is 16, 17, 18, 19, 20).

[0024] In some embodiments, a is 2, b is 1 or 2, and c is 16, 18, or 20.

[0025] In some embodiments, X 10 is Y1, Y1 is Lys having a substituent at the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH, where a is 2, b is 1 or 2, and c is 16 or 18.

[0026] In some embodiments, X 12 is Y1, Y1 is Lys having a substituent at the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH, where a is 2, b is 1 or 2, and c is 16 or 18.

[0027] In some embodiments, X 13 is Y1, Y1 is Lys having a substituent at the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH, where a is 2, b is 1 or 2, and c is 16 or 18.

[0028] In some embodiments, X 14 is Y1, Y1 is Lys having a substituent at the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH, where a is 2, b is 1 or 2, and c is 16 or 18.

[0029] In some embodiments, X 16is Y1, Y1 is Lys having a substituent at the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH, where a is 2, b is 1 or 2, and c is 16 or 18.

[0030] In some embodiments, X 17 is Y1, Y1 is Lys having a substituent at the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH, where a is 2, b is 1 or 2, and c is 16 or 18.

[0031] In some embodiments, the side chain amino group of a Lys residue in Y1 is covalently linked to the substituent by forming an amide bond.

[0032] In some embodiments, Y is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the structure: [ka] K(-OEG-OEG-yGlu-C20-OH) has the following structure: [ka] .

[0033] In some embodiments, Y is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein: K(-OEG-OEG-yGlu-C18-OH) has the following structure: [ka] K(-OEG-OEG-yGlu-C20-OH) has the following structure: [ka] .

[0034] In some embodiments, the ε-amino group of the Lys residue in Y1 is covalently linked to the substituent via an amide bond and the α-amino group of the Lys residue is linked to the peptide chain.

[0035] In some embodiments, X 1 is chosen from Tyr, and X 2 is chosen from Aib, and X 10 is chosen from Tyr, and X 12 is chosen from Ile, and X 13 is chosen from Tyr, and X 14 is selected from Y1, and X 15 is selected from Asp or Glu, and X 16 is selected from Arg or Lys, and X 17 is chosen from Ile, and X 18 is chosen from Ala, and X 19 is chosen from Ala, and X 20 is selected from Gln, and X 23 is selected from Ile or Val, and X 24 is selected from Asn, and X 27 is selected from Ile or Leu, and X 28 is chosen from Ala, and X 29 is selected from Gly, and X 30 is selected from Gly, and Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the following structure: [ka] K(-OEG-OEG-yGlu-C20-OH) has the following structure: [ka] .

[0036] In some embodiments, X 1 is chosen from Tyr, and X 2 is chosen from Aib, and X 10 is chosen from Tyr, and X 12 is chosen from Ile, and X 13 is chosen from Tyr, and X 14 is selected from Y1, and X 15 is selected from Asp or Glu, and X 16 is selected from Arg or Lys, and X 17 is chosen from Ile, and X 18 is chosen from Ala, and X 19 is chosen from Ala, and X 20 is selected from Gln, and X 23 is selected from Ile or Val, and X 24 is selected from Asn, and X 27 is selected from Ile or Leu, and X 28 is chosen from Ala, and X 29 is selected from Gly, and X 30 is selected from Gly, and Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the following structure: [ka] K(-OEG-OEG-yGlu-C20-OH) has the following structure: [ka] .

[0037] In some embodiments of the present disclosure, the GLP-1 analog is shown in general formula (II) (SEQ ID NO: 20): H-YAibEGTFTSDYSIYX 14 X 15 X 16 IAAQEFX 23 NWLX 27AGGPSSGAPPPS-NH 2 (II), of which X 14 is K or L, X 15 D, E, X 16 is K, R, X 23 is V or I, X 27 is I or L.

[0038] In some embodiments of the present disclosure, the GLP-1 analog is selected from the compounds numbered 1 to 18 below: TIFF2024546026000009.tif186155

[0039] In some embodiments of the present disclosure, the GLP-1 analog is selected from the compounds shown below in numbers 1# to 18#: TIFF2024546026000010.tif186163

[0040] In some embodiments, the GLP-1 analog of the present disclosure is selected from compounds shown in Figure 3 as 7#, 12#, 13#, 14#, 15#, 16#, 17#, or 18#.

[0041] In some embodiments, the concentration of GLP-1 analogs in the composition is 0.01mg / mL~1000mg / mL, for example 0.1mg / mL~500mg / mL, 0.1mg / mL~400mg / mL, 0.1mg / mL~300mg / mL, 0.1mg / mL~300mg / mL mg / mL~200mg / mL、0.1mg / mL~100mg / mL、0.5mg / mL~200mg / mL、0.5mg / mL~150mg / mL、0.5mg / mL~100mg / mL、0.5mg / mL~50mg / mL、0.5mg / mL~25mg / mL、1.0mg / mL g / mL~100mg / mL, 1.0mg / mL~90mg / mL, 1.0mg / mL~80mg / mL, 1.0mg / mL~70mg / mL, 1.0mg / mL~60mg / mL, 1.0mg / mL~50mg / mL, 1.0mg / mL~40mg / mL, 1.0mg / mL~30mg / mL, 1.0mg / mL~20mg / mL, 1.0mg / mL~10mg / mL, 1.0mg / mL~9.0mg / mL, 1.0mg / mL~8.0mg / mL, 1.0mg / mL~7.0mg / mL, 1.0mg / mL~6.0mg / mL, 1.0mg / mL~5. 0mg / mL, 2.0mg / mL~50mg / mL, 2.0mg / mL~40mg / mL, 2.0mg / mL~30mg / mL, 2.0mg / mL~20mg / mL, 2.0mg / mL~10mg / mL, 2.0mg / mL~9.0mg / mL, 2.0mg / mL~8.0mg / mL, 2.0mg / mL~7.0mg / mL, 2.0mg / mL~6.0mg / mL, 2.0mg / mL~5.0mg / mL, 5.0mg / mL~1000mg / mL, 5.0mg / mL~500mg / mL, 5.0mg / mL~400mg / mL, 5.0mg / mL~300m g / mL、5.0mg / mL~200mg / mL、5.0mg / mL~100mg / mL、5.0mg / mL~90mg / mL、5.0mg / mL~80mg / mL、5.0mg / mL~70mg / mL、5.0mg / mL~60mg / mL、5.0mg / mL~50mg / mL 、5.0mg / mL~40mg / mL、5.0mg / mL~30mg / mL、5.0mg / mL~20mg / mL、5.0mg / mL~10mg / mL、6.0mg / mL~1000mg / mL、6.0mg / mL~500mg / mL、6.0mg / mL~400mg / mL、6.0mg / mL~300mg / mL, 6.0mg / mL~200mg / mL, 6.0mg / mL~100mg / mL, 6.0mg / mL~90mg / mL, 6.0mg / mL~80mg / mL, 6.0mg / mL~70mg / mL, 6. 0mg / mL~60mg / mL, 6.0mg / mL~50mg / mL, 6.0mg / mL~40mg / mL, 6.0mg / mL~30mg / mL, 6.0mg / mL~20mg / mL, or 6.0mg / mL~10mg / mL. In some embodiments, the concentration of the GLP-1 analog or a medicamentable salt thereof in the pharmaceutical composition is about 1.0 mg / mL, about 2.0 mg / mL, about 3.0 mg / mL, about 4.0 mg / mL, about 5.0 mg / mL, about 6.0 mg / mL, about 7.0 mg / mL, about 8.0 mg / mL, about 9.0 mg / mL, about 10.0 mg / mL, about 11.0 mg / mL, about 12.0 mg / mL, about 13.0 mg / mL, about 14.0 mg / mL, about 15.0 mg / mL, about 16.0 mg / mL, about 17.0 mg / mL, about 18.0 mg / mL, about 19.0 mg / mL, about 20.0 mg / mL, about 21.0 mg / mL, about 22.0 mg / mL, about 23.0 mg / mL, about 24.0 mg / mL, about 25.0 mg / mL, about 26.0 mg / mL, about 27.0 mg / mL, about 28.0 mg / mL, about 29.0 mg / mL, about 30.0 mg / mL, about 31.0 mg / mL, about 32.0 mg / mL, about 33.0 mg / mL, about 34.0 mg / mL, about 35.0 mg / mL, about 36.0 mg / mL, about 37.0 mg / mL, about 38.0 mg / mL, about 39.0 mg / mL, about 40.0 mg / mL, about 41.0 mg / mL, about 42.0 mg / mL, about 43.0 mg / mL, about 44.0 mg / mL, about 45.0 mg / mL, about 46.0 mg / mL, about 47.0 mg / mL, about 48.0 mg / mL, about 49.0 mg / mL, about 5 / mL, about 15.0 mg / mL, about 16.0 mg / mL, about 17.0 mg / mL, about 18.0 mg / mL, about 19.0 mg / mL, about 20.0 mg / mL, about 21.0 mg / mL, about 22.0 mg / mL, about 23.0 mg / mL, about 24.0 mg / mL, about 25.0 mg / mL, about 26.0 mg / mL, about 27.0 mg / mL, about 28.0 mg / mL, about 29.0 mg / mL, about 30.0 mg / mL. In some embodiments, the concentration of the GLP-1 analog or a medicamentable salt thereof in the pharmaceutical composition is about 2.0 mg / mL or about 10.0 mg / mL.

[0042] In some embodiments, the pharmaceutical composition further comprises an osmolality modifier, including, but not limited to, salts (e.g., sodium chloride, phosphates, sodium citrate, boric acid, and sodium tartrate), sugars or sugar alcohols (lactose, trehalose, sucrose, glucose, mannitol, sorbitol, xylitol), amino acids (e.g., L-glycine, L-histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine), polyhydroxy sugar alcohols (e.g., glycerin, 1,2-propylene glycol (also referred to as propylene glycol in this disclosure), 1,3-propylene glycol, 1,3-butanediol), polyethylene glycols (e.g., PEG 400), or mixtures thereof.

[0043] In some embodiments, the penetration modifier is one or more selected from propylene glycol, mannitol, sorbitol, xylitol, glycerin, lactose, trehalose, sucrose, glucose, sodium chloride, phosphates, sodium citrate, boric acid, and sodium tartrate.

[0044] In some embodiments, the osmotic adjuster is sodium chloride.

[0045] In some embodiments, the osmolarity adjusting agent is propylene glycol or mannitol.

[0046] In some embodiments, the concentration of propylene glycol in the pharmaceutical composition is 10 mg / mL to 20 mg / mL, for example, 10 mg / mL to 19 mg / mL, 10 mg / mL to 18 mg / mL, 10 mg / mL to 17 mg / mL, 10 mg / mL to 16 mg / mL, 10 mg / mL to 15 mg / mL, 11 mg / mL to 20 mg / mL, 11 mg / mL to 19 mg / mL, 11 mg / mL to 18 mg / mL, 11 mg / mL to 17 mg / mL, 11 mg / mL to 16 mg / mL, mL, 11 mg / mL to 15 mg / mL, 12 mg / mL to 20 mg / mL, 12 mg / mL to 19 mg / mL, 12 mg / mL to 18 mg / mL, 12 mg / mL to 17 mg / mL, 12 mg / mL to 16 mg / mL, 12 mg / mL to 15 mg / mL, 13 mg / mL to 20 mg / mL, 13 mg / mL to 19 mg / mL, 13 mg / mL to 18 mg / mL, 13 mg / mL to 17 mg / mL, 13 mg / mL to 16 mg / mL, 13 mg / mL to 15 mg / mL. In some embodiments, the concentration of propylene glycol in the pharmaceutical composition is 12 mg / mL to 16 mg / mL, for example about 14 mg / mL.

[0047] In some embodiments, the concentration of sodium chloride in the pharmaceutical composition is 1 mg / mL to 20 mg / mL, for example, 1 mg / mL to 19 mg / mL, 1 mg / mL to 18 mg / mL, 1 mg / mL to 17 mg / mL, 1 mg / mL to 16 mg / mL, 1 mg / mL to 15 mg / mL, 2 mg / mL to 18 mg / mL, 2 mg / mL to 17 mg / mL, 2 mg / mL to 16 mg / mL, 2 mg / mL to 15 mg / mL, 3 mg / mL to 18 mg / mL, 3 mg / mL to 17 mg / mL, 3 mg / mL to 16 mg / mL, 3 mg / mL to 15 mg / mL, 4 mg / mL to 14 mg / mL, 5 mg / mL to 13 mg / mL, 6 mg / mL to 12 mg / mL, 7 mg / mL to 15 mg / mL, 8 mg / mL to 15 mg / mL, 9 mg / mL to 12 mg / mL, 10 mg / mL to 15 mg / mL, 11 mg / mL to 15 mg / mL, 12 mg / mL to 15 mg / mL, 13 mg / mL to 15 mg / mL, 14 mg / mL to 15 mg / mL, 15 mg / mL to 15 mg / mL, 16 mg / mL to 15 mg / mL, 17 mg / mL to 15 mg / mL, 18 mg / mL to 15 mg / mL, 17 mg / mL to 15 mg / mL, 18 mg / mL to 15 mg / mL, 19 mg / mL to 20 mg / mL, 20 mg / mL to 25 mg / mL, 21 mg / mL to 25 mg / mL, 22 mg / mL to 25 mg / mL, 23 mg / mL to 25 mg / mL, 24 mg / mL to 25 In some embodiments, the concentration of sodium chloride in the pharmaceutical composition is 2 mg / mL to 18 mg / mL. In some embodiments, the concentration of sodium chloride in the pharmaceutical composition is from 3 mg / mL to 15 mg / mL. In some embodiments, the concentration of sodium chloride in the pharmaceutical composition is 7 mg / mL to 10 mg / mL, e.g., about 7.5 mg / mL, about 7.6 mg / mL, about 7.7 mg / mL, about 7.8 mg / mL, about 7.9 mg / mL, about 8 mg / mL, about 8.1 mg / mL, about 8.2 mg / mL, about 8.3 mg / mL, about 8.4 mg / mL, about 8.5 mg / mL, about 8.6 mg / mL, about 8.7 mg / mL, about 8.8 mg / mL, about 8.9 mg / mL, about 9.0 mg / mL, about 9.1 mg / mL, about 9.28 mg / mL, about 9.3 mg / mL, about 9.4 mg / mL, about 9.5 mg / mL, about 9.6 mg / mL, about 9.7 mg / mL, about 9.8 mg / mL, about 9.9 mg / mL, or about 10 mg / mL.In some embodiments, the concentration of sodium chloride in the pharmaceutical composition is 7.5 mg / mL to 9.5 mg / mL.

[0048] In some embodiments, the concentration of mannitol in the pharmaceutical composition is 10 mg / mL to 50 mg / mL, for example, 15 mg / mL to 45 mg / mL, 20 mg / mL to 45 mg / mL, 25 mg / mL to 45 mg / mL, 30 mg / mL to 45 mg / mL, 15 mg / mL to 40 mg / mL, 15 mg / mL to 35 mg / mL, 20 mg / mL to 40 mg / mL, 20 mg / mL to 35 mg / mL, 25 mg / mL to 40 mg / mL, 25 mg / mL to 35 mg / mL, 30 mg / mL to 40 mg / mL, or 35 mg / mL to 40 mg / mL. In some embodiments, the concentration of mannitol in the pharmaceutical composition is about 25 mg / mL, 25.5 mg / mL, about 26 mg / mL, 26.5 mg / mL, about 27 mg / mL, 27.5 mg / mL, about 28 mg / mL, 28.5 mg / mL, about 29 mg / mL, 29.5 mg / mL, about 30 mg / mL, 30.5 mg / mL, about 31 mg / mL, 31.5 mg / mL, about 32 mg / mL, 32.5 mg / mL, about 33 mg / mL, 33.5 mg / mL, about 34 mg / mL, 34.5 mg / mL, about 35 mg / mL, 35.5 mg / mL, about 36 mg / mL, 36.5 mg / mL, about 37 mg / mL, 37.5 mg / mL, about 38 mg / mL, 38.5 mg / mL, about 39 mg / mL, 39.5 mg / mL, or about 40 mg / mL.

[0049] In some embodiments, the concentration of the buffer in the pharmaceutical composition is 0.5 mM to 50.0 mM, e.g., 0.5 mM to 40.0 mM, 0.5 mM to 30.0 mM, 0.5 mM to 20.0 mM, 0.5 mM to 10.0 mM, 1.0 mM to 40.0 mM, 1.0 mM to 35.0 mM, 1.0 mM to 30.0 mM, 1.0 mM to 25.0 mM, , 1.0 mM to 20.0 mM, 1.0 mM to 15.0 mM, 1.0 mM to 10.0 mM, 2.0 mM to 40.0 mM, 2.0 mM to 35.0 mM, 2.0 mM to 30.0 mM, 2.0 mM to 25.0 mM, 2.0 mM to 20.0 mM, 2.0 mM to 15.0 mM, 2.0 mM to 10.0 mM, or 5.0 mM to 10.0 mM. In some embodiments, the concentration of the buffer in the pharmaceutical composition is about 1.0 mM, about 2.0 mM, about 3.0 mM, about 4.0 mM, about 5.0 mM, about 6.0 mM, about 7.0 mM, about 8.0 mM, about 9.0 mM, or about 10.0 mM. In some embodiments, the concentration of the buffering agent in the pharmaceutical composition is about 5.0 mM.

[0050] In some embodiments, the pharmaceutical composition further comprises a pH adjusting agent, such as sodium hydroxide and / or hydrochloric acid.

[0051] In some embodiments, the pH of the pharmaceutical composition is 6.5 to 9.0, for example, 6.6 to 9.0, 6.7 to 9.0, 6.8 to 9.0, 6.9 to 9.0, 7.0 to 9.0, 7.1 to 9.0, 7.2 to 9.0, 7.3 to 9.0, 7.4 to 9.0, 7.5 to 9.0, 7.6 to 9.0, 7.7 to 9.0, 7.8 to 9.0, 7.9 to 9.0, 8.0 to 9.0, 7.0 to 8.9, 7.0 to 8.8, 7.0 to 8.7, 7.0 to 8.6, 7.0 to 8.5, 7.0 to 8.4, 7.0 to 8.3, 7.0 to 8.2, 7.0 to 8.1, 7.0 to 8.0, 7.1 to 8.9, 7.1 to 8.8, 7.1 to 8.7, 7.1 to 8.6, 7.1 to 8.5, 7.1 to 8.4, 7.1 to 8.3, 7.1 to 8.2, 7.1 to 8.1, 7.1 to 8.0, 7.1 to 7.9, 7.1 to 7.8, or 7.1 to 7.7. In some embodiments, the pH of the pharmaceutical composition is about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, or about 8.5. In some embodiments, the pH of the pharmaceutical composition is 7.0 to 8.0. In some embodiments, the pH of the pharmaceutical composition is 7.1 to 7.7, for example, about 7.5 or about 7.4.

[0052] In some embodiments, the pharmaceutical composition further comprises a pharma- ceutically acceptable bacteriostatic agent.

[0053] In some embodiments, pharma- ceutically acceptable bacteriostatic agents include, but are not limited to, phenol, o-cresol, m-cresol, p-cresol, methylparaben, propylparaben, 2-phenoxyethanol, butylparaben, 2-phenylethyl alcohol, benzyl alcohol, ethanol, chlorobutanol, and thimerosal, bromopropylene glycol, benzoic acid, imidurea, chlorhexidine, sodium dehydroacetate, chlorocresol, ethylparaben, benzethonium chloride, or mixtures thereof.

[0054] In some embodiments, the pharma- ceutically acceptable bacteriostatic agent is phenol.

[0055] In some embodiments, the concentration of the bacteriostatic agent in the pharmaceutical composition is 4.0 mg / mL to 7.0 mg / mL, for example, 4.2 mg / mL to 6.9 mg / mL, 4.4 mg / mL to 6.9 mg / mL, 4.6 mg / mL to 6.9 mg / mL, 4.8 mg / mL to 6.9 mg / mL, 5.0 mg / mL to 6.9 mg / mL, 5.1 mg / mL to 6.9 mg / mL, 5.2 mg / mL to 6.9 mg / mL, 5.3 mg / mL to 6.9 mg / mL, 5.4 mg / mL to 6.9 mg / mL, 5.5 mg / mL to 6.9 mg / mL, 5.6 mg / mL to 6.9 mg / mL, 5.7 mg / mL to 6.9 mg / mL, 5.8 mg / mL to 6.9 mg / mL, 5.9 ... L~6.9mg / mL, 5.4mg / mL~6.9mg / mL, 5.5mg / mL~6.9mg / mL, 4.2mg / mL~6.8mg / mL, 4.4mg / mL~6.8mg / mL, 4.6mg / mL~ 6.8mg / mL, 4.8mg / mL~6.8mg / mL, 5.0mg / mL~6.8mg / mL, 5.1mg / mL~6.8mg / mL, 5.2mg / mL~6.8mg / mL, 5.3mg / mL~6.8 mg / mL, 5.4mg / mL~6.8mg / mL, 5.5mg / mL~6.8mg / mL, 4.4mg / mL~6.7mg / mL, 4.6mg / mL~6.7mg / mL, 4.8mg / mL~6.7mg / mL, 5.0mg / mL~6.7mg / mL, 5.1mg / mL~6.7mg / mL, 5.2mg / mL~6.7mg / mL, 5.3mg / mL~6.7mg / mL, 5.4mg / mL~6.7mg / m L, 5.5 mg / mL to 6.7 mg / mL, 4.4 mg / mL to 6.6 mg / mL, 4.6 mg / mL to 6.6 mg / mL, 4.8 mg / mL to 6.6 mg / mL, 5.0 mg / mL to 6.6 mg / mL, 5.1 mg / mL to 6.6 mg / mL, 5.2 mg / mL to 6.6 mg / mL, 5.3 mg / mL to 6.6 mg / mL, 5.4 mg / mL to 6.6 mg / mL, 5.5 mg / mL to 6.6 mg / mL. In some embodiments, the concentration of the bacteriostatic agent is 4.2 mg / mL to 6.9 mg / mL. In some embodiments, the concentration of the bacteriostatic agent is 5.5 mg / mL to 6.6 mg / mL, for example about 5.5 mg / mL.

[0056] In some embodiments, the pharmaceutical composition comprises any one of 1) to 6) below: 1) The compound shown in FIG. 3 #18 or a medicinal salt thereof; Buffers, e.g., phosphate buffers such as sodium dihydrogen phosphate; propylene glycol or mannitol, and an optional antimicrobial agent such as phenol; 2) The compound shown in FIG. 3 #18 or a medicinal salt thereof; Buffers, e.g., phosphate buffers such as sodium dihydrogen phosphate; Sodium chloride, and an optional antimicrobial agent such as phenol; 3) The compound shown in FIG. 3 #18 or a medicinal salt thereof; Buffers, e.g., phosphate buffers such as sodium dihydrogen phosphate; propylene glycol or mannitol, an optional antimicrobial agent such as phenol; and a pH adjuster such as sodium hydroxide and / or hydrochloric acid; 4) The compound shown in FIG. 3 #18 or a medicinal salt thereof; Buffers, e.g., phosphate buffers such as sodium dihydrogen phosphate; Sodium chloride, an optional antimicrobial agent such as phenol; and a pH adjuster such as sodium hydroxide and / or hydrochloric acid; 5) The compound shown in FIG. 3 #18 or a medicinal salt thereof; Buffers, e.g., phosphate buffers such as sodium dihydrogen phosphate; propylene glycol or mannitol, an optional antimicrobial agent such as phenol; pH adjusters such as sodium hydroxide and / or hydrochloric acid, As well as water for injection, 6) The compound shown in FIG. 3 #18 or a medicinal salt thereof; Buffers, e.g., phosphate buffers such as sodium dihydrogen phosphate; Sodium chloride, an optional antimicrobial agent such as phenol; pH adjusters such as sodium hydroxide and / or hydrochloric acid, And water for injection.

[0057] In some embodiments, the pharmaceutical composition consists of the compound shown in 18# of FIG. 3 or a medicamentable salt thereof, sodium dihydrogen phosphate, propylene glycol or mannitol, phenol, sodium hydroxide and / or hydrochloric acid, and water for injection.

[0058] In some embodiments, the pharmaceutical composition consists of the compound shown in 18# of FIG. 3 or a medicamentable salt thereof, sodium dihydrogen phosphate, propylene glycol or mannitol, sodium hydroxide and / or hydrochloric acid, and water for injection.

[0059] In some embodiments, the pharmaceutical composition consists of the compound shown in FIG. 3 at 18# or a medicamentable salt thereof, sodium dihydrogen phosphate, sodium chloride, phenol, sodium hydroxide and / or hydrochloric acid, and water for injection.

[0060] In some embodiments, the pharmaceutical composition consists of the compound shown in FIG. 3 at 18# or a medicamentable salt thereof, sodium dihydrogen phosphate, sodium chloride, sodium hydroxide and / or hydrochloric acid, and water for injection.

[0061] In some embodiments, the pharmaceutical composition comprises any one of A-K: A) 1.0 mg / mL to 100 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate, and 10 mg / mL to 20 mg / mL propylene glycol or 10 mg / mL to 50 mg / mL mannitol, And the pH of the pharmaceutical composition is 6.5 to 9.0.

[0062] B) 1.0 mg / mL to 100 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate, and Sodium chloride from 2mg / mL to 18mg / mL, And the pH of the pharmaceutical composition is 6.5 to 9.0.

[0063] C) 1.0 mg / mL to 100 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate, 10 mg / mL to 20 mg / mL propylene glycol, or 15 mg / mL to 45 mg / mL mannitol, or 2 mg / mL to 18 mg / mL sodium chloride, An antimicrobial agent of choice, such as phenol at 4.0 mg / mL to 7.0 mg / mL; And the pH of the pharmaceutical composition is 6.5 to 9.0.

[0064] D) 1.0 mg / mL to 30.0 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 25.0 mM sodium dihydrogen phosphate, 11 mg / mL to 18 mg / mL propylene glycol, or 20 mg / mL to 40 mg / mL mannitol, or 3 mg / mL to 15 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.2mg / mL to 6.9mg / mL; And the pH of the pharmaceutical composition is 7.0 to 8.0.

[0065] E) 2.0 mg / mL to 10.0 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 2.0 mM to 10.0 mM sodium dihydrogen phosphate, 12 mg / mL to 16 mg / mL propylene glycol, or 25 mg / mL to 35 mg / mL mannitol, or 8 mg / mL to 10 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.4mg / mL to 6.8mg / mL; And the pH of the pharmaceutical composition is 7.1 to 7.7.

[0066] F) 1.0 mg / mL to 100 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate, 10 mg / mL to 20 mg / mL propylene glycol, or 15 mg / mL to 45 mg / mL mannitol, or 2 mg / mL to 18 mg / mL sodium chloride, An antimicrobial agent of choice, such as phenol at 4.0 mg / mL to 7.0 mg / mL; As well as water for injection, And the pH of the pharmaceutical composition is 6.5 to 9.0.

[0067] G) 1.0 mg / mL to 30.0 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 25.0 mM sodium dihydrogen phosphate, 11 mg / mL to 18 mg / mL propylene glycol, or 20 mg / mL to 40 mg / mL mannitol, or 3 mg / mL to 15 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.2mg / mL to 6.9mg / mL; As well as water for injection, And the pH of the pharmaceutical composition is 7.0 to 8.0.

[0068] H) 2.0 mg / mL to 20.0 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 2.0 mM to 10.0 mM sodium dihydrogen phosphate, 12 mg / mL to 16 mg / mL propylene glycol, or 25 mg / mL to 35 mg / mL mannitol, or 7 mg / mL to 10 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.4mg / mL to 6.8mg / mL; As well as water for injection, And the pH of the pharmaceutical composition is 7.1 to 7.7.

[0069] I) 5.0 mg / mL to 15.0 mg / mL or 5.0 mg / mL to 10.0 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 5.0 mM to 10.0 mM sodium dihydrogen phosphate, 12 mg / mL to 16 mg / mL propylene glycol, or 7 mg / mL to 10 mg / mL sodium chloride, or 8 mg / mL to 9 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.4mg / mL to 6.8mg / mL; As well as water for injection, And the pH of the pharmaceutical composition is 7.1 to 7.7.

[0070] J) 5.0 mg / mL to 15.0 mg / mL or 5.0 mg / mL to 10.0 mg / mL of the compound shown in 18# of Figure 3 or a medicinal salt thereof; 4.0 mM to 8.0 mM or 4.0 mM to 6.0 mM sodium dihydrogen phosphate, 12 mg / mL to 16 mg / mL propylene glycol, or 7 mg / mL to 10 mg / mL sodium chloride, or 8 mg / mL to 9 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.4mg / mL to 6.8mg / mL; As well as water for injection, And the pH of the pharmaceutical composition is 7.1 to 7.7.

[0071] K) 1.0 mg / mL to 20.0 mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 1.0 mM to 25.0 mM sodium dihydrogen phosphate, 11 mg / mL to 18 mg / mL propylene glycol, or 20 mg / mL to 40 mg / mL mannitol, or 3 mg / mL to 15 mg / mL sodium chloride, Selective antibacterial agents such as phenol at 4.2mg / mL to 6.9mg / mL; And the pH of the pharmaceutical composition is 7.0 to 8.0.

[0072] In some embodiments, the pharmaceutical composition comprises: 6.0mg / mL to 10.0mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 2.0 mM to 10.0 mM sodium dihydrogen phosphate, and Contains 30mg / mL to 40mg / mL of mannitol.

[0073] In some embodiments, the pharmaceutical composition comprises: 6.0mg / mL to 10.0mg / mL of the compound shown in FIG. 3 #18 or a medicinal salt thereof; 2.0 mM to 10.0 mM sodium dihydrogen phosphate, and glycerin, the concentration of the glycerin being 10 to 30, for example, 10, 20 or 30 mg.

[0074] In some embodiments, the disclosure provides a pharmaceutical composition comprising: (1) A pharmaceutical composition comprising about 2.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (2) A pharmaceutical composition comprising about 4.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (3) A pharmaceutical composition comprising about 5.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (4) A pharmaceutical composition comprising about 6.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (5) A pharmaceutical composition comprising about 8.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (6) A pharmaceutical composition comprising about 10.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (7) A pharmaceutical composition comprising about 20.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (8) A pharmaceutical composition comprising about 2.0 mg / mL, about 4.0 mg / mL, about 5.0 mg / mL, about 6.0 mg / mL, about 8.0 mg / mL, 10 mg / mL or about 20 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 8 to 9 mg / mL (e.g., about 8.2 mg / mL) of sodium chloride, and optionally about 5.5 mg / mL of phenol, and having a pH of about 7.5 or about 7.4; (9) The pharmaceutical compositions according to (1) to (8), wherein sodium dihydrogen phosphate is replaced from 5.0 mM to 10.0 mM. (10) The pharmaceutical composition according to (1) to (9), wherein the osmotic pressure adjusting agent is replaced with glycerin having a concentration of, for example, 20 mg / kg; or (11) The pharmaceutical composition according to any one of (1) to (10), wherein the composition is adjusted to 1 mL with water for injection when a fixed volume is required, and the final volume is adjusted to 1 mL.

[0075] In some embodiments, the disclosure provides a pharmaceutical composition comprising: (1) A pharmaceutical composition comprising about 2.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, about 5.5 mg / mL of phenol, a suitable amount of sodium hydroxide and / or a suitable amount of hydrochloric acid, and the remainder of water for injection, and having a pH of about 7.5 or about 7.4; (2) A pharmaceutical composition comprising about 4.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, about 5.5 mg / mL of phenol, a suitable amount of sodium hydroxide and / or a suitable amount of hydrochloric acid, and the remainder of water for injection, and having a pH of about 7.5 or about 7.4; (3) A pharmaceutical composition comprising about 5.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, about 5.5 mg / mL of phenol, a suitable amount of sodium hydroxide and / or a suitable amount of hydrochloric acid, and the remainder of water for injection, and having a pH of about 7.5 or about 7.4; (4) A pharmaceutical composition comprising about 6.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, about 5.5 mg / mL of phenol, a suitable amount of sodium hydroxide and / or a suitable amount of hydrochloric acid, and the remainder of water for injection, and having a pH of about 7.5 or about 7.4; (5) A pharmaceutical composition comprising about 8.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, about 5.5 mg / mL of phenol, a suitable amount of sodium hydroxide and / or a suitable amount of hydrochloric acid, and the remainder of water for injection, and having a pH of about 7.5 or about 7.4; (6) A pharmaceutical composition comprising 10.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or 31.5 mg / mL of mannitol, 5.5 mg / mL of phenol, an appropriate amount of sodium hydroxide and / or an appropriate amount of hydrochloric acid, and the remainder of water for injection, and having a pH of 7.5 or about 7.4; (7) A pharmaceutical composition comprising 20.0 mg / mL of the compound shown in FIG. 3 #18 or a medicament salt thereof, 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or 31.5 mg / mL of mannitol, 5.5 mg / mL of phenol, an appropriate amount of sodium hydroxide and / or an appropriate amount of hydrochloric acid, and the remainder of water for injection, and having a pH of 7.5 or about 7.4; (8) A pharmaceutical composition comprising about 2.0 mg / mL, about 4.0 mg / mL, about 5.0 mg / mL, about 6.0 mg / mL, about 8.0 mg / mL, about 10 mg / mL or about 20 mg / mL of the compound shown in 18# of FIG. 3 or a medicinal salt thereof, about 5.0 mM sodium dihydrogen phosphate, 8 to 9 mg / mL (e.g., about 8.2 mg / mL) of sodium chloride, and optionally about 5.5 mg / mL of phenol, an appropriate amount of sodium hydroxide and / or an appropriate amount of hydrochloric acid, and the remainder of water for injection, and having a pH of about 7.5 or about 7.4; (9) The pharmaceutical compositions according to (1) to (8), wherein sodium dihydrogen phosphate is replaced from 5.0 mM to 10.0 mM. (10) The pharmaceutical composition according to (1) to (9), wherein the osmotic pressure adjusting agent is replaced with glycerin having a concentration of, for example, 20 mg / kg; or (11) The pharmaceutical composition according to any one of (1) to (10), wherein the composition is adjusted to 1 mL with water for injection when a fixed volume is required, and the final volume is adjusted to 1 mL.

[0076] The pharmaceutical composition of the present disclosure already has sufficient drug stability and can be left stably for a long period of time.

[0077] In some embodiments, the pharmaceutical composition is stable for at least 3 months, at least 6 months, at least 12 months, at least 18 months, at least 24 months, or at least 36 months at 2° C.-8° C. In some embodiments, the pharmaceutical composition is stable for at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months at 25° C. In some embodiments, the pharmaceutical composition is stable for at least 7 days, at least 14 days, at least 28 days, at least 1 month, at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months at 40° C.

[0078] The present disclosure provides a method for preparing the above pharmaceutical composition, comprising the step of dissolving the GLP-1 analogue or a medicamentable salt thereof.

[0079] For ease of drug delivery, the pharmaceutical compositions of the present disclosure can further be prepared as lyophilized formulations.

[0080] The present disclosure provides a lyophilized formulation, which, after being reconstituted, can form any one of the pharmaceutical compositions described above.

[0081] The present disclosure further provides a method for preparing a lyophilized formulation comprising a GLP-1 analog or a medicament salt thereof, comprising lyophilizing the pharmaceutical composition. In an alternative embodiment, the lyophilization comprises, in sequence, the steps of pre-freezing, primary drying and secondary drying.

[0082] The present disclosure further provides a lyophilized formulation comprising a GLP-1 analogue or a medicinal salt thereof, prepared by the above-described method for preparing a lyophilized formulation comprising a GLP-1 analogue or a medicinal salt thereof.

[0083] The present disclosure further provides a method for preparing a reconstituted solution of a lyophilized formulation comprising a GLP-1 analogue or a medicamentable salt thereof, comprising a step of reconstituting said lyophilized formulation, wherein the solution used for the reconstitution is selected from, but not limited to, water for injection, saline or glucose solution.

[0084] The present disclosure further provides a reconstituted solution of a freeze-dried formulation containing a GLP-1 analog or a medicinal salt thereof, obtained by the above-mentioned method for preparing a reconstituted solution of a freeze-dried formulation containing a GLP-1 analog or a medicinal salt thereof.

[0085] The disclosure further provides an article of manufacture or reagent kit comprising a container housing any of the stable pharmaceutical compositions described herein, in some embodiments, the glass vial is an injection bottle made of neutral borosilicate glass tubing.

[0086] The present disclosure further provides a product comprising a container containing the pharmaceutical composition or the lyophilized formulation or a reconstitution solution of the lyophilized formulation.

[0087] The present disclosure further provides a pharmaceutical composition or a lyophilized formulation or a reconstituted solution of a lyophilized formulation for use in a method of treating or preventing a disease or condition.

[0088] The present disclosure further provides the use of the above pharmaceutical composition or a lyophilized formulation or a reconstituted solution of a lyophilized formulation in the preparation of a medicament for treating and / or preventing a disease or condition.

[0089] The present disclosure further provides a method for treating and preventing a disease or condition, comprising administering to a patient in need thereof a therapeutically effective amount of the above-described pharmaceutical composition or lyophilized formulation or a reconstituted solution of the lyophilized formulation.

[0090] The present disclosure provides for the use of a pharmaceutical composition in the preparation of a medicament for treating non-insulin dependent diabetes mellitus, insulin dependent diabetes mellitus, obesity, non-alcoholic fatty liver disease, hepatic steatosis, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, insulin resistance, dyslipidemia associated with insulin resistance, and / or dyslipidemia associated with diabetes.

[0091] The present disclosure provides pharmaceutical compositions for treating non-insulin dependent diabetes mellitus / type II diabetes mellitus, insulin dependent diabetes mellitus, obesity, non-alcoholic fatty liver disease, hepatic steatosis, insulin resistance associated dyslipidemia, and / or dyslipidemia associated with diabetes.

[0092] The present disclosure provides a method for treating non-insulin dependent diabetes mellitus / type II diabetes, insulin dependent diabetes mellitus, obesity, non-alcoholic fatty liver disease, hepatic steatosis, insulin resistance associated dyslipidemia, and / or diabetes associated dyslipidemia comprising administering to a subject in need thereof a pharmaceutical composition of the present disclosure.

[0093] The present disclosure provides compounds that can simultaneously activate the GLP-1 receptor and the GIP receptor, and in some embodiments, the GLP-1 analogs have greater agonist activity at the GLP-1R than at the GIP receptor.

[0094] In some specific embodiments, the GLP-1 analogs of the present disclosure have a ratio of agonist activity at the GLP-1R to agonist activity at the GIP receptor of (1-10):1, (1.1-10):1, (1.1-9.5):1, (1.1-9):1, (1.1-8.5):1, (1.1-8):1, (1.1-7.5):1, (1.1-7):1, (1.1-6.5):1, (1.1-6):1, (1.2-10):1, (1.2-9.5):1, (1.2-9):1, (1.2-8.5):1, (1.2-8):1, (1.2-7.5):1, ( 1.2~7):1,(1.2~6.5):1,(1.2~6):1,(1.3~10):1,(1.3~9.5):1,(1.3~9):1,(1.3~8.5):1,(1.3~8):1,(1.3~7.5):1,(1.3~7):1,(1.3~6.5):1,(1.3~6):1,(1.4~10):1,(1.4~9.5):1,(1.4~9):1,(1.4~8.5):1,(1.4~8):1,(1.4~7.5):1,(1.4~7):1,(1.4~6.5):1,(1.4~6):1,(1.5~10):1,(1.5~9 .5):1,(1.5~9):1,(1.5~8.5):1,(1.5~8):1,(1.5~7.5):1,(1.5~7):1,(1.5~6.5):1,(1.5~6):1,(2~10):1,(2~9.5):1,(2~9):1,(2~8.5):1,(2~8):1,(2~7.5):1,(2~7):1,(2~6.5):1,(2~6):1,(2.5~10):1,(2.5~9.5):1,(2.5~9):1,(2.5~8.5):1,(2.5~8):1,(2.5~7.5):1,(2.5~7):1,(2.5~6 .5):1, (2.5~6):1, (3~10):1, (3~9.5):1, (3~9):1, (3~8.5):1, (3~8):1, (3~7.5):1, (3~7):1, (3~6.5):1, (3~6):1, (3.5~10):1, (3.5~9.5):1, (3.5~9):1, (3.5~8.5):1, (3.5~8):1, (3.5~7.5):1, (3.5~7):1, (3.5~6.5):1, (3.5~6):1, (4~10):1, (4~9.5):1, (4~9):1, (4~8.5):1, (4~8):1, (4~7.5):1,(4~7):1,(4~6.5):1,(4~6):1,(4.5~10):1,(4.5~9.5):1,(4.5~9):1,(4.5~8.5):1,(4.5~8):1,(4.5~7.5):1,(4.5~7):1,(4.5~6.5):1,(4.5~6):1,(5~10):1,(5~9.5):1,(5~9):1,(5~8.5):1,(5~8):1,(5~7.5):1,(5~7):1,(5~6.5):1,(5~6):1,(5~5.5):1,(5 .1-5.5):1, (5.2-5.4):1, (5.2-5.3):1 or any range or point therebetween, such as about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 2:1, about 2.5:1, about 3:1, about 3.5:1, about 4:1, about 4.5:1, about 5:1, about 5.2:1, about 5.3:1, about 5.4:1, about 5.5:1, about 6:1, about 6.5:1, about 7:1, about 7.5:1, about 8:1, about 8.5:1, about 9:1, about 9.5:1, about 10:1. The above ratios are normalized to data from the corresponding in vitro agonist activity assay. For example, the corresponding agonist activity may be measured with a cAMP-Gs kinetics reagent kit. In this context, the expressions (1-10):1 and 1:1-10:1 have the same meaning.

[0095] In another embodiment, the present disclosure provides the above GLP-1 analogs and their pharma- ceutically acceptable salts. The GLP-1 analogs provided by the present disclosure belong to amphoteric compounds that can exhibit both acidic and basic properties. Those skilled in the art can use acidic or basic compounds to react with the GLP-1 analogs provided by the present disclosure to form salts by known techniques.

[0096] Pharmaceutical compositions containing GLP-1 analogs according to the present disclosure can be used to treat patients in need of such treatment by parenteral administration. The parenteral administration route can be selected as subcutaneous injection, intramuscular injection or intravenous injection. The polypeptide dual agonist compounds of the present disclosure can further be selected for administration by transdermal route, can be selected for iontophoretic patch, or can be selected for administration by transmucosal route.

[0097] The GLP-1 analogues provided by the present disclosure are synthesized by the method of solid phase synthesis. In one example, the synthesis vector is Rink-amide MBHA (Xi'an Lanxiao Technology) resin. During synthesis, the α-amino group of the amino acid derivative used is protected with Fmoc group. For example, the side chains of amino acids are protected with a protecting group selected from the following depending on the functional group: the sulfhydryl group of the cysteine ​​side chain, the amino group of the asparagine and glutamine side chains, and the imidazolyl group of the histidine side chain are protected with Trt (trityl group), the guanidine group of the arginine side chain is protected with Pbf (2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl group), the indolyl group of the tryptophan side chain and the amino group of the lysine side chain are protected with Boc (tert-butoxycarbonyl group), and the carboxyl group of the aspartic acid and glutamic acid side chains, the hydroxyl group of the threonine side chain, the phenol group of the tyrosine side chain, and the hydroxyl group of the serine side chain are protected with t-Bu (tert-butyl group). For example, during synthesis, the carboxy group of the C-terminal amino acid residue of the polypeptide is first condensed to the polymeric insoluble Rink-amide MBHA resin in the form of an amide bond, and then the Fmoc protecting group at the α-amino group is removed with a N,N-dimethylformamide (DMF) solution containing 20% ​​4-methylpiperidine, and in excess, the solid-phase vector is condensed with the next amino acid derivative in the polypeptide sequence to form an amide bond to extend the peptide chain. The length of the synthesized polypeptide chain is obtained by repeating the operation of "condensation → washing → deprotection → washing → next amino acid condensation", and finally the polypeptide is decomposed from the solid-phase vector by reacting the resin with a mixed solution of trifluoroacetic acid: water: triisopropylsilane (for example, 90: 5: 5, v: v: v), and then precipitated with 5 volumes of frozen methyl tert-butyl ether to obtain a solid crude product of GLP-1 analog. The solid crude polypeptide product is dissolved in a mixed solution of acetonitrile / water containing 0.1% trifluoroacetic acid, and then purified and separated on a C-18 reverse phase preparative chromatography column to obtain a pure GLP-1 analog.

[0098] According to some embodiments, the present disclosure provides a method for producing a cellular membrane comprising: a GLP-1 analogue according to the present disclosure or a pharma- ceutically acceptable salt thereof; -Another therapeutic agent selected from any one or a combination of an anti-obesity agent, an anti-diabetic agent, an anti-hypertensive agent, and a lipid-lowering agent; Further provided is a pharmaceutical kit-of-parts comprising: wherein the GLP-1 analog or a pharma- ceutically acceptable salt thereof is in a separate container from the other therapeutic agent. In some embodiments, the GLP-1 analog or a pharma- ceutically acceptable salt thereof is administered to a subject separately or in combination with the other therapeutic agent (e.g., administered simultaneously or sequentially).

[0099] In some embodiments, the pharmaceutical composition of the present disclosure is provided in combination with an administration device (e.g., a syringe, injection pen, or auto-injector). As one example, the pharmaceutical composition of the present disclosure is pre-loaded into an administration device so that a subject can self-administer it at home. As another example, the pharmaceutical composition of the present disclosure is provided separately from the administration device. [Brief description of the drawings]

[0100] [Figure 1] 1 shows the effect of compounds of the present disclosure on the rate of weight change in diet-induced obese mice. [Diagram 2] FIG. 1 shows the effect of compounds of the present disclosure on daily food intake in diet-induced obese mice. [Diagram 3] 1 shows structures of exemplary compounds of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0101] In order that the present disclosure may be more readily understood, certain technical and scientific terms are specifically defined below. Unless expressly defined otherwise herein, all other technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art.

[0102] The amino acid sequences of the present disclosure include the standard one-letter or three-letter codes for the 20 amino acids, and unless otherwise indicated, the preferred configuration for all amino acid residues in the present disclosure is L-, where Aib is alpha aminoisobutyric acid, D-Ala is D-alanine, Orn is ornithine, Dap is 2,3-diaminopropionic acid, and Dab is 2,4-diaminobutyric acid.

[0103] The term "agonist" is defined as a substance that has an activating effect on the GLP-1 receptor or the GIP receptor.

[0104] The term "GLP-1 / GIP dual agonist" as used in the context of the present disclosure refers to a substance or ligand capable of activating the GLP-1 receptor and the GIP receptor.

[0105] In this disclosure, the term "treatment" includes inhibiting, alleviating, halting, or reversing the progression or severity of an existing condition or disease.

[0106] "Natural amino acids" are the 20 common amino acids, namely, alanine (A), cysteine ​​(C), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), proline (P), glutamine (Q), arginine (R), serine (S), threonine (T), valine (V), tryptophan (W), and tyrosine (Y).

[0107] "Unnatural amino acids" are non-naturally encoded amino acids, or amino acids that are not expressed in the genetic code of any organism. For example, unnatural amino acids may be purely synthetic compounds. Illustrative examples of unnatural amino acids are hydroxyproline, γ-carboxyglutamic acid, O-phosphoserine, azetidine carboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid, β-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, tert-butylglycine, 2,4-diaminoisobutyric acid (Dap), desmosine, 2,2'-diaminopimelic acid, 2,3-diaminopropionic acid (Dab), N-ethylglycine, N-methylglycine, N- Examples of amino acids include, but are not limited to, ethylasparagine, pipecolic acid, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, alloisoleucine, N-methylalanine, N-methylglycine, N-methylisoleucine, N-methylpentylglycine, N-methylvaline, naphthylalanine, norvaline, norleucine, ornithine (Orn), D-ornithine, D-arginine, p-aminophenylalanine, pentylglycine, pipecolic acid, and thioproline. The term also includes derivatives obtained by chemically modifying the C-terminal carboxyl group (or the N-terminal amino group and / or its side chain functional group) of a natural amino acid (or an unnatural amino acid).

[0108] The term "alkyl group" refers to a saturated aliphatic hydrocarbon group that is a straight or branched chain group containing from 1 to 20 carbon atoms, such as an alkyl group containing from 1 to 8 carbon atoms, such as an alkyl group containing from 1 to 6 carbon atoms, such as an alkyl group containing from 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 6-methylhexyl, 7-methylhexyl, 8-methylhexyl, 9-methylhexyl, 10-methylhexyl, 11-methylhexyl, 12-methylhexyl, 13-methylhexyl, 14-methylhexyl, 15-methylhexyl, 16-methylhexyl, 17-methylhexyl, 18-methylhexyl, 19-methylhexyl, 20-methylhexyl, 21-methylhexyl, 22-methylhexyl, 23-methylhexyl, 24-methylhexyl, 25-methylhexyl, 26-methylhexyl, 27-methylhexyl, 28-methylhexyl, 29-methylhexyl, 30-methylhexyl, 31-methylhexyl, 32-methylhexyl, 33-methylhexyl, 34-methylhexyl, 35-methylhexyl, 36-methylhexyl, 37-methylhexyl, 38-methylhexyl, 39-methylhexyl, 40-methylhexyl, 41-methylhexyl, 42-methylhexyl, 43-methylhex Examples of such groups include 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof.An alkyl group can be, for example, a lower alkyl group containing from 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any accessible attachment site, and may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy, or carboxylic acid ester groups. The substituted alkyl groups of the present disclosure may be methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkyl substituted with an alkoxy group, or alkyl substituted with a hydroxy group.

[0109] In the present disclosure, different terms such as "X is selected from A, B, or C", "X is selected from A, B, and C", "X is A, B, or C", and "X is A, B, and C" all mean the same thing, i.e., X may be any one or more of A, B, and C.

[0110] An amino acid "modification" as described in this disclosure means an amino acid substitution, addition or deletion, including the substitution or addition of any one or more of the 20 naturally occurring amino acids.

[0111] The term "native GLP-1" refers to a naturally occurring molecule of the glucagon or exendin family of peptides, which are encoded by the proglucagon gene and include three small highly homologous peptides, glucagon(1-29), GLP-1(1-37), and GLP-2(1-33), and exendins are peptides expressed in lizards that are similar to GLP-1 and stimulate insulin secretion. In some embodiments, the term "native GLP-1" also refers to human GLP-1(7-37) and human GLP-1(7-36).

[0112] The term "GLP-1 analogue" refers to having 25 or less, 24 or less, 23 or less, 22 or less, 21 or less, 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, 15 or less, 14 or less, 13 or less, 12 or less, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 amino acid or chemical modification compared to native GLP-1 (particularly compared to human GLP-1(7-37) and human GLP-1(7-36)), said amino acid modification may be an amino acid substitution, addition and / or deletion, said chemical modification may be a chemical modification selected from amides, carbohydrates, alkyl groups, acyl groups, esters, polyethylene glycol (PEG) groups, sialyl groups, glycosylation groups, and the like.

[0113] An amino acid "substitution" as described in this disclosure refers to the replacement of one amino acid residue with another amino acid residue.

[0114] The term "polyethylene glycol" or "PEG" refers to a mixture of polycondensates of ethylene oxide and water, which exists in linear or branched form and has the general formula H(OCH 2 CH2 ) n OH, where n is greater than or equal to 9. Unless further explained, the term includes polymers of polyethylene glycol having a total average molecular weight selected from 5,000 daltons to 40,000 daltons.

[0115] The term "fatty acid" refers to a carboxylic acid having a long chain fatty acid tail, which may be saturated or unsaturated, and in this disclosure fatty acids are carboxylic acids having a C4 to C30 straight or branched aliphatic group.

[0116] The term "peptide" as used in this disclosure covers the range of peptides having modified amino and carboxy termini, for example, an amino acid chain in which the terminal carboxylic acid is replaced with an amide group is included within the amino acid sequence named for natural amino acids.

[0117] Any hydrogen atom described in the present disclosure may be replaced with its isotope (protium, deuterium, tritium), and any hydrogen atom in the compounds of the present disclosure according to the present disclosure may be replaced with an isotope atom.

[0118] "Optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the phrase includes cases where the event or circumstance occurs and cases where the event or circumstance does not occur. For example, "a heterocyclyl group optionally substituted with an alkyl group" means that the alkyl group may or may not be present, and the description includes cases where the heterocyclyl group is substituted with an alkyl group and cases where the heterocyclyl group is not substituted with an alkyl group.

[0119] "Substituted" means that one or more hydrogen atoms in the group, preferably 5 or less, more preferably 1 to 3 hydrogen atoms, are substituted with a substituent independently of each other. Substituents are located only at their chemically possible sites, and a person skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino group or a hydroxy group having a free hydrogen may be unstable when bonded to a carbon atom having an unsaturated (e.g., olefinic) bond.

[0120] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein or their physiologically / pharmaceutical acceptable salts or prodrugs and other chemical components, such as physiologically / pharmaceutical acceptable vectors, excipients, etc. The pharmaceutical composition is intended to facilitate administration to a living body and contribute to the absorption of the active ingredient to further exert biological activity.

[0121] "Agonist activity" refers to the ability of a compound according to the present disclosure to activate the human GIP receptor and the human GLP-1 receptor. In some examples, "agonist activity" is expressed in terms of relative activity, specifically, the ratio of the ability of a compound according to the present disclosure to activate the GLP-1R to the GIP receptor.

[0122] "Pharmaceutically acceptable salt" refers to a salt of a compound according to the present disclosure, which is safe and effective when used in the mammalian body and has the desired biological activity. Such salts are generally acid addition salts or base salts. Acid addition salts include inorganic acid salts and organic acid salts.

[0123] Semaglutide is a once-weekly GLP-1 receptor single agonist polypeptide drug developed by Novo Nordisk of Denmark, and is currently approved and commercially available in the United States, Japan and the European Union.

[0124] LY3298176 refers to a once-weekly GIP receptor / GLP-1 receptor dual agonist polypeptide drug developed by Eli Lilly and Company, which is currently undergoing Phase III clinical trials in multiple countries. The structure is as follows:

[0125] TIFF2024546026000011.tif9158, of which the following fatty acids [ka] is modified at the 20th K.

[0126] "Buffer" refers to a buffer that resists changes in pH by the action of its acid-base conjugate components. Examples of buffers that control pH in the appropriate range include acetate, succinate, citrate, phosphate, gluconate, histidine, oxalate, lactate, phosphate, citrate, tartrate, fumarate, glycylglycine and other organic acid buffers.

[0127] A "histidine salt buffer" is a buffer containing histidine ions. Examples of histidine salt buffers include histidine-hydrochloride, histidine-acetate, histidine-phosphate, histidine-sulfate, and the like, such as histidine-acetate buffer or histidine-hydrochloride buffer, where histidine-acetate buffer is prepared from histidine and acetic acid, and histidine salt buffer is prepared from histidine and hydrochloric acid.

[0128] A "citrate buffer" is a buffer that contains citrate ions. Examples of citrate buffers include citric acid-sodium citrate, citric acid-potassium citrate, citric acid-calcium citrate, citric acid-magnesium citrate, etc. The citrate buffer may be citrate-sodium citrate.

[0129] A "succinate buffer" is a buffer that contains succinate ions. Examples of succinate buffers include succinic acid-sodium succinate, succinic acid-potassium succinate, succinic acid-calcium succinate, etc. The succinate buffer may be succinic acid-sodium succinate.

[0130] A "phosphate buffer" is a buffer that contains phosphate ions. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate, disodium hydrogen phosphate-citric acid, etc. The phosphate buffer may be disodium hydrogen phosphate-sodium dihydrogen phosphate.

[0131] An "acetate buffer" is a buffer that contains acetate ions. Examples of acetate buffers include acetic acid-sodium acetate, acetic acid histidine salt, acetic acid-potassium acetate, acetic acid-calcium acetate, acetic acid-magnesium acetate, etc. The acetate buffer may be acetic acid-sodium acetate.

[0132] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein or their physiologically / pharmaceutical acceptable salts or prodrugs, and other chemical components such as physiologically / pharmaceutical acceptable vectors and excipients. The pharmaceutical composition is intended to facilitate administration to a living body and contribute to the absorption of the active ingredient to further exert biological activity. In this application, the terms "pharmaceutical composition" and "formulation" may be used interchangeably.

[0133] In the solution forms of the pharmaceutical compositions described in this disclosure, unless otherwise specified, the solvent therein is water.

[0134] "Lyophilized formulation" refers to a pharmaceutical composition or formulation obtained after the step of freeze-drying under vacuum a liquid or solution form of the pharmaceutical composition or a solution formulation.

[0135] The term "about" or "approximately" as used herein means that a numerical value is within an acceptable error range of a specific value as determined by one of ordinary skill in the art, the numerical portion being determined by how it is measured or determined (i.e., the limitations of the measurement system). For example, in the practice of the art, "about" may mean within or more than 1 standard deviation. Alternatively, "about" or "essentially including" may mean a range of at most ±20%, e.g., a pH of about 5.5 means pH 5.5 ±1.1. Moreover, particularly for biological systems or processes, the term may mean at most an order of magnitude or at most 5 times the numerical value. Unless otherwise indicated, when a specific value appears in the present application and claims, the meaning of "about" or "essentially including" should be assumed to be within an acceptable error range of the specific value.

[0136] The pharmaceutical compositions described in the present disclosure can achieve a stable effect, i.e., the GLP-1 analog therein essentially retains its physical and / or chemical stability and / or biological activity after storage, e.g., the pharmaceutical composition essentially retains its physical and chemical stability and its biological activity after storage. The storage period is generally selected based on the desired shelf life of the pharmaceutical composition. Currently, there are several analytical techniques for measuring protein stability, which can measure stability after storage at a given temperature for a given period of time.

[0137] A stable drug formulation is one that does not show significant changes when stored at refrigerated temperatures (2°C-8°C) for at least 3 months, at least 6 months, at least 1 year, at least 2 years, and even up to 2 years. Stable liquid formulations also include liquid formulations that exhibit the desired characteristics after storage at temperatures including 25°C for periods including 1 month, 3 months, 6 months, or at 40°C for periods including 1 month. A typical acceptable criterion for stability is that, as measured by SEC-HPLC, typically no more than about 10%, e.g., no more than about 5%, of the GLP-1 analog monomer is degraded. Upon visual analysis, the drug formulation is colorless or clear to slightly opalescent. The concentration, pH, and osmolality of the formulation have a variation of no more than ±10%. Typically no more than about 10%, e.g., no more than about 5%, cleavage is observed, and typically no more than about 10%, e.g., no more than about 5% aggregates are formed.

[0138] A GLP-1 analog "retains its physical stability" in a drug formulation if it does not exhibit significant increased aggregation, precipitation, and / or denaturation as determined by visual inspection of color and / or clarity or by UV light scattering, size exclusion chromatography (SEC) and dynamic light scattering (DLS). Changes in protein conformation may be assessed by fluorescence spectroscopy (which determines the tertiary structure of a protein) and by FTIR spectroscopy (which determines the secondary structure of a protein).

[0139] If the GLP-1 analogue does not show any significant chemical changes, it is said that the GLP-1 analogue "retains its chemical stability" in the drug formulation. Chemical stability can be evaluated by detecting and quantifying chemically altered proteins. Degradation processes that constantly change the chemical structure of proteins include hydrolysis or cleavage (assessed by methods such as size exclusion chromatography and SDS-PAGE), oxidation (assessed by methods such as peptide mapping combined with mass spectrometry or MALDI / TOF / MS), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, measurement of isoaspartic acid, etc.) and isomerization (assessed by measurement of isoaspartic acid content, peptide mapping, etc.).

[0140] A GLP-1 analog "retains its biological activity" in a drug formulation if the biological activity of the GLP-1 analog over a given time period is within a given range of the biological activity exhibited when the drug formulation was prepared. The biological activity of a GLP-1 analog can be determined, for example, by antigen binding assays.

[0141] The three-letter and one-letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968). EXAMPLES

[0142] In order to describe the present disclosure in more detail, the present specification provides the following embodiments for carrying out the present disclosure, but the embodiments of the present disclosure are not limited thereto. Experimental methods for which specific conditions are not specified in the examples of the present disclosure generally follow conventional conditions or conditions recommended by manufacturers of raw materials or products. Reagents for which a specific source is not specified are ordinary reagents that are commercially available.

[0143] [Table 1]

[0144] [Table 2]

[0145] Example 1. Chemical synthesis of compound 18# 1. Synthesis of the polypeptide backbone Rink-amide MBHA resin (substitution degree: 0.48 mmol / g, 0.1 mmol) was weighed and placed in a polypropylene polypeptide synthesis solid-phase reaction tube, N,N-dimethylformamide (DMF, 10 mL) was added and the resin was swelled under nitrogen-blowing for 10 minutes, DMF was removed by vacuum, DMF (10 mL) was added to wash the resin, and the resin washing was repeated twice, after which the synthesis of the polypeptide solid phase was carried out by the Fmoc / tBu strategy on a Prelude-X polypeptide fully automated synthesizer, among which 10 equivalents of amino acid residues activated with HCTU and 4-methylmorpholine (HCTU, 4-methylmorpholine, amino acid residue molar ratio is 1:2:1) were used, and the coupling was realized by condensing the amide bond by reacting in DMF at room temperature for 25 minutes. The N-terminal Fmoc protecting group was deprotected by reacting twice (10 minutes each time) at room temperature using a DMF solution containing 20% ​​4-methylpiperidine. During the synthesis of the polypeptide backbone, the N-terminal amino acid residue was selected to be constructed as Boc-L-Tyr(tBu)-OH and a second condensation was carried out, which is necessary to improve the quality of the crude peptide.

[0146] 2. Selective deprotection of the resin peptide protecting group Mtt ​​and modification of side chain fatty acid After completion of the elongation of the above polypeptide backbone (also called resin peptide), a mixed solution of dichloromethane containing 30% hexafluoroisopropanol (10 mL) was added, and the mixture was reacted by shaking at room temperature for 45 minutes, after which the mixed solution was removed. A mixed solution of dichloromethane containing 30% hexafluoroisopropanol (10 mL) was further added, and the mixture was reacted by shaking at room temperature for 45 minutes, after which the mixed solution was removed. After completion of the reaction, the resin was washed six times with DMF. The 14th lysine side chain was extended using a Prelude-X fully automated polypeptide synthesizer, and an additional coupling / deprotection cycle was performed to obtain the amino acid building block Fmoc-NH-PEG. 2 The N-terminal Fmoc protecting group was deprotected by reacting twice at room temperature for 10 min each time using a DMF solution containing 20% ​​4-methylpiperidine. Finally, the resin obtained was washed three times with DCM and DMF, and then 10 equivalents of HOOC-(CH 2 ) 18 A DMF mixture (8 mL) containing -COOtBu, 10 equivalents of HCTU, and 20 equivalents of diisopropylethylamine (DIEA) was added, and the reaction was carried out at room temperature for 4 hours to complete the modification of the fatty acid in the side chain.

[0147] 3. Decomposition of products The resin peptide obtained in the previous step was washed three times with DMF and DCM, then dried in vacuum, and then the newly prepared decomposition solution (trifluoroacetic acid: triisopropylsilane: water = 90: 5: 5, volume ratio) was added and the mixture was shaken at room temperature for 3 to 4 hours to react. After the reaction was completed, the mixture was filtered, the resin was washed twice with trifluoroacetic acid, and the filtrate was combined and a large amount of frozen methyl tert-butyl ether was added to precipitate the solid. After centrifugation, the supernatant was removed to obtain the crude polypeptide product of compound 18#.

[0148] 4. Purification by reverse phase liquid chromatography The crude polypeptide product of compound 18# was dissolved in a mixed solvent containing 0.1% trifluoroacetic acid, 20% acetonitrile, and 20% acetic acid / water, filtered through a 0.22 μm film, and separated by a WATERS Prep150 LC reversed-phase high-performance liquid chromatography system, with buffers A (0.1% trifluoroacetic acid, 10% acetonitrile, aqueous solution) and B (0.1% trifluoroacetic acid, 90% acetonitrile, aqueous solution). Among them, the chromatography column was an X-SELECT OBD C-18 reversed-phase chromatography column, and the detection wavelength of the chromatograph during purification was set to 220 nm, and the flow rate was 15 mL / min. The distillate related to the product was collected and lyophilized to obtain the pure polypeptide product of compound 1#, with a yield of 18%. The purity of the pure polypeptide product was confirmed by analytical high-performance liquid chromatography and ultra-high-performance liquid chromatography / mass spectrometry, of which the purity was 92.81%. Molecular structure of compound 18#: TIFF2024546026000015.tif10145, the structural formula is shown in Figure 3, structure 18#.

[0149] Example 2. Chemical synthesis of other compounds The compounds in Table 3 were synthesized according to the experimental design of Example 1: [Table 3]

[0150] The purity of the compounds was determined by analytical HPLC and ultra-HPLC / mass spectrometry, and the purity of some compounds is shown in Table 4 below: [Table 4]

[0151] Biological Testing and Evaluation The present disclosure will be further described below in conjunction with test examples, but these examples are not intended to limit the scope of the present disclosure.

[0152] Example 3. Evaluation of the agonist activity of compounds according to the present disclosure against the glucagon-like peptide-1 receptor (GLP-1R)

[0153] 1. Purpose of the experiment: The purpose of this test example is to measure the agonist activity of the compounds according to the present disclosure against the glucagon-like peptide-1 receptor (GLP-1R).

[0154] 2. Experimental method: The frozen CHO-K1 / GLP-1R / CRE-luc stable transformed cell line (which can be prepared by a common method in this field) is taken out of the liquid nitrogen container, put into a water bath at 37°C to rapidly melt, resuspended in DMEM / F12 medium, centrifuged, washed once, resuspended in DMEM / F12 medium containing 0.1% casein as an experimental buffer, adjusted the cell density with the experimental buffer, seeded in a 384-well plate (Sigma Cat# CLS4514) at a density of 2500 cells / 5μL / well, and added 2.5μL of IBMX working solution (Sigma Cat# I7018) with a final concentration of 0.5mM IBMX prepared with buffer and 2.5μL of gradient diluted polypeptide sample to each well, centrifuged at 1000rpm for 1 minute, shaken for 30 seconds to mix evenly, and left to incubate at room temperature for 30 minutes. Detected with Cisbio cAMP-Gs Dynamic kit (Cisbio Cat#62AM4PEC), and cAMP-d2 and Anti-cAMP-Eu 3+ -Cryptate was diluted 20-fold with cAMP Lysis & Detection Buffer and mixed evenly. 5 μL of diluted cAMP-d2 solution was added to each well, and then diluted Anti-cAMP-Eu 3+ 5 μL of -Cryptate solution was added, the mixture was shaken for 30 seconds to mix evenly, and the mixture was incubated at room temperature in the dark for 1 hour.

[0155] 3. Experimental data processing method: The HTRF signal was read using a Biotek Synergy H1 microplate reader, with an excitation wavelength of 320 nm and emission wavelengths of 620 nm and 665 nm. The signal ratio (665 nm / 620 nm × 10,000) was calculated, and the signal ratio and sample concentration were nonlinearly fitted with a four-variable equation in GraphPad Prism 6 to obtain the EC 50 The values ​​obtained are shown in Table 5 below.

[0156] Example 4. Evaluation of the agonist activity of compounds according to the present disclosure at the glucose-dependent insulinotropic peptide receptor (GIPR)

[0157] 1. Purpose of the experiment: The compounds were tested for agonist activity at the glucose-dependent insulinotropic peptide receptor (GIPR).

[0158] 2. Experimental method: Wild-type CHO-K1 cells were harvested, the cell suspension was adjusted to an appropriate density, and seeded into 6-well plates at 2 mL per well and incubated at 37°C, 5% CO 2The cells were placed in an incubator to attach to the wall overnight, and the transfection mixture (hGIP receptor plasmid, Fugene HD (Promega Cat# E2311), OptiMEM (Gibco Cat# 31985070) was mixed evenly and left at room temperature for 15 minutes, then added to the corresponding cell wells in a volume of 100 μL, and transfected for 24 hours to overexpress hGIP receptor on the CHO-K1 cell surface. After the transient transfection was completed, the cells in the 6-well plate were collected and washed once with DMEM / F12 medium (Gibco Cat# 11330032) containing 0.1% casein (Sigma Cat# C3400) as the experimental buffer, and the cell density was adjusted with the experimental buffer, and seeded into a 384-well plate (Sigma Cat# CLS4514) at a density of 5000 cells / 5 μL / well, and then each well was filled with IBMX working solution (Sigma Cat# CLS4514) with a final concentration of 0.5 mM IBMX prepared in the buffer. 2.5μL of Anti-cAMP-Eu3+-Cryptate solution was added to each well, and 2.5μL of gradient diluted polypeptide sample was added. The mixture was centrifuged at 1000rpm for 1 minute, shaken for 30 seconds to mix evenly, and incubated at room temperature for 30 minutes. Detection was performed using Cisbio cAMP-Gs Dynamic kit (Cisbio Cat# 62 AM4PEC). cAMP-d2 and Anti-cAMP-Eu3+-Cryptate were diluted 20-fold with cAMP Lysis & Detection Buffer and mixed evenly. 5μL of diluted cAMP-d2 solution was added to each well, and then 5μL of diluted Anti-cAMP-Eu3+-Cryptate solution was added, shaken for 30 seconds to mix evenly, and incubated at room temperature for 1 hour in the dark.

[0159] 3. Experimental data processing method: The HTRF signal was read using a Biotek Synergy H1 microplate reader, with an excitation wavelength of 320 nm and emission wavelengths of 620 nm and 665 nm. The signal ratio (665 nm / 620 nm × 10,000) was calculated, and the signal ratio and sample concentration were nonlinearly fitted with a four-variable equation in GraphPad Prism 6 to obtain the EC 50The values ​​obtained are shown in Tables 5 and 6 below.

[0160] [Table 5]

[0161] [Table 6]

[0162] 4. Experimental conclusion: By designing the polypeptide backbone and subsequent site-specific modification of fatty acids, the compounds according to the present disclosure have stronger GLP-1 / GIPR agonist activity than many GLP-1 / GIPR dual agonist polypeptides in the field, and therefore have better potential for treating metabolic diseases. Furthermore, unlike LY3298176, which shows preferential activity against GIPR, compounds 12#-18# according to the present disclosure show preferential activity against GLP-1R.

[0163] Example 5. Stability testing of some compounds according to the present disclosure Plasma stability is important for therapeutic polypeptide drugs because they are likely to be sensitive to polypeptide hydrolases and proteases in plasma. Unstable polypeptides in plasma will affect their half-life and therapeutic efficacy.

[0164] 1. Purpose of the experiment: This experiment is aimed at testing the stability of some compounds according to the present disclosure in human plasma.

[0165] 2. Experimental method: 5μL of samples with concentrations of 20, 50, 100, 200, 500, 1000, 2000, 5000, 10000ng / mL were added to 45μL of human plasma, and the content of the compounds therein was detected by LC-MS to create a standard curve. 5μL of a polypeptide solution with a concentration of 1mg / mL was added to 45μL of human plasma. Five samples were prepared for each test compound, and one sample was taken out after 0 minutes, 30 minutes, 60 minutes, 120 minutes, and 240 minutes, and the content of the compounds remaining therein was detected by LC-MS, and the relative content of the compounds remaining in the samples at other times was calculated based on the time at 0 minutes (100%). The compound detection method by LC-MS is to prepare 5% acetonitrile solution as solution A, and 95% acetonitrile solution as solution B, and form a solution gradient at a flow rate of 0.6mL / min with the time and solution ratio shown in Table 9 below, inject 15μL of sample, and detect the compound content by Raptor Biphenyl 2.7 micron detection column, see Table 7.

[0166] [Table 7]

[0167] 3. Experimental results: Plasma stability data for some compounds of the present disclosure are shown in Table 8 below.

[0168] [Table 8]

[0169] Test conclusion: Studies have revealed that compound 7# according to the present disclosure has similar stability in human plasma at 4 hours compared to compound LY3298176 (relative content >90%).

[0170] Example 6. Pharmacokinetic properties of some compounds according to the present disclosure in mice Plasma stability is one of the factors that affect the pharmacokinetics of polypeptide drugs, which is further influenced by factors such as their absorption and clearance in the body.

[0171] 1. Purpose of the experiment: The purpose of this experiment is to study the pharmacokinetic behavior in the mouse body (plasma) of a compound according to the present disclosure following a single intravenous injection using Balb / c mice as test animals.

[0172] 2. Experimental method: Male Balb / c mice weighing 18g-30g and aged 7-9 weeks were purchased from Shanghai Jiesijie Laboratory Animal Co.,Ltd. Compound 7# was prepared in a buffer solution (pH=7.0) containing 20mM citric acid, and then compound 7# was injected into the mouse body through the tail vein at a dose of 30nmol / kg body weight, and 0.2mL of blood was collected at 0 hours, 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 24 hours, and 32 hours. At a temperature of 4°C, the collected mouse blood was centrifuged at a speed of 6000 rpm for 6 minutes to separate plasma. The content of compound 7# in mouse plasma was detected according to the experimental method of Example 3.3.

[0173] 3. Experimental results: According to the above experimental method, the specific data are shown in Table 9 below: [Table 9]

[0174] 4. Experimental conclusion: Research has shown that compound 7# of the present disclosure has good pharmacokinetic properties after intravenous administration in mice, which shows advantages in treating diseases, for example, it can support weekly subcutaneous administration in humans.

[0175] Example 7. Pharmacokinetic properties of some compounds according to the present disclosure in mice

[0176] 1. Purpose of the experiment: The purpose of this experiment is to study the pharmacokinetic behavior in mice (plasma) following a single subcutaneous injection of a compound according to the present disclosure using Balb / c mice as test animals.

[0177] 2. Experimental method: Male Balb / c mice weighing 18g-30g and aged 7-9 weeks were purchased from Shanghai Jiesijie Laboratory Animal Co., Ltd. Compound 7# was prepared in a buffer solution (pH=7.0) containing 20mM citric acid, and then compound 7# was injected subcutaneously into the left flank of the mouse at a dose of 30nmol / kg body weight. 0.2mL of blood was collected at 0 hours, 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 24 hours, and 32 hours. At a temperature of 4℃, the collected mouse blood was centrifuged at a speed of 6000rpm for 6 minutes to separate plasma. The content of compound 7# in mouse plasma was detected according to the experimental method of Example 5.2.

[0178] 3. Experimental results: According to the above experimental method, the specific data are shown in Table 10 below: [Table 10]

[0179] 4. Experimental conclusion: Studies have shown that the compounds disclosed herein have good pharmacokinetic properties after subcutaneous administration in mice, which may be advantageous in treating diseases, for example, supporting weekly subcutaneous administration in humans.

[0180] Example 8. In vivo efficacy of some compounds according to the present disclosure

[0181] 1. Purpose of the experiment: The regulatory effect of some of the disclosed compounds and the compound LY3298176 on blood glucose in normal mice following a single subcutaneous administration is tested.

[0182] 2. Experimental method: Male C57BL / 6 mice aged 10-12 weeks were purchased from Shanghai Jieshijie Laboratory Animal Co., Ltd. C57BL / 6 mice were subcutaneously injected with compound 7# or compound LY3298176 (dose: 10 nmol / kg body weight) and control buffer, then fasted but not deprived of water, and 18 hours later, a glucose solution with a concentration of 0.2 g / mL was injected via the abdominal cavity. According to the experimental design, blood was collected from the tail of the mouse at 0 minutes, 15 minutes, 30 minutes, 60 minutes, and 120 minutes, and the blood glucose level was measured. The specific method was to fix the mouse by physical means, expose the tail, cut off a small part of the tail, press the tail to bleed, discard the first drop of blood, and detect blood glucose with a Roche Active blood glucose meter. The area under the blood glucose curve (AUC) was calculated based on the results at each time point.

[0183] 3. Experimental results: According to the above experimental method, the specific data are shown in Table 11 below: [Table 11]

[0184] 4. Experimental conclusion: In this experiment, at a dose of 10 nmol / kg body weight, compound 7# of the present disclosure exhibited a significant hypoglycemic effect in normal mice, with the area under the blood glucose curve in the compound 7# group being reduced by more than 60% compared with that in the placebo (i.e., blank solvent).

[0185] Example 9. Weight-reducing effects of some compounds according to the present disclosure

[0186] 1. Purpose of the experiment: The numbered compounds are tested for their modulating effect on body weight in diet-induced obese mice by subcutaneous administration.

[0187] 2. Experimental method: Obese male C57BL / 6 mice (weight 35g-55g, age 10-12 weeks, purchased from Shanghai Jieshijie Laboratory Animal Co., Ltd.) induced by high fat diet were subcutaneously injected with compounds LY3298176 (10nmol / kg body weight), 7# (10nmol / kg body weight), and 18# (three doses, 3nmol / kg, 10nmol / kg, and 100nmol / kg body weight, respectively, administered once every 3 days). According to the experimental design, the weight of each mouse was weighed and recorded on days 0, 3, 6, and so on until day 27, and the average weight of the mice in each group was calculated, and the weight change curve was drawn based on the weight on the first day. After the study, the fat and other organs of the mice were removed and weighed, and the fat / brain ratio of each part of each mouse was calculated. The effects of the drug on fat were determined by comparing the changes in fat, organ / brain ratios of different parts of the mice in each group.

[0188] 3. Experimental results: According to the above experimental method, the specific data are shown in Tables 12 to 14 and FIG. 1 below: [Table 12]

[0189] 4. Experimental conclusion: In this experiment, at doses of 3 nmol / kg, 10 nmol / kg and 100 nmol / kg, compounds 7# and 18# of the present disclosure exhibited significant weight-reducing effects on high-fat diet-induced obese mice, and showed significant dose-related effects. Compound 18# reduced the body weight of the 10 nmol / kg test group by more than 20.0% on the 27th day, while control compound LY3298176 reduced the body weight of the test group by about 13.4% under the same dose conditions. In addition, the fat content (except scapular fat) of each site of the test groups of different doses of compound 18# was significantly reduced compared to the placebo (i.e., blank solvent) group. [Table 13]

[0190] [Table 14]

[0191] [Table 15]

[0192] Example 10. Effect of compounds of the present disclosure on food intake in mice Food intake of the mice in each group was measured daily during the study, and the results are shown in Table 1 and FIG.

[0193] The model control group DIO (diet-induced obesity) mice had an average daily food intake of 2.5 g during the entire study, and after different doses of compound 18# or LY3298176 were subcutaneously injected, the food intake of the mice in each group decreased to different degrees.

[0194] On the first day after administration, the food intake of the mice in each treatment group was significantly reduced. The food intake of the mice in the 3 nmol / kg, 10 nmol / kg, and 100 nmol / kg compound 18# groups was 0.6 g, 0.3 g, and 0.2 g, respectively, which was significantly different from the model control group (2.5 g) and showed a relatively good dose-response relationship.

[0195] The mice in the model control group had a cumulative food intake of 12.8 g over 5 days after administration, while the mice in the compound 18# groups administered 3 nmol / kg, 10 nmol / kg, and 100 nmol / kg had cumulative food intakes of 7.2 g, 3.9 g, and 1.8 g over 5 days after administration, which were significantly lower than those of the model control group and showed a relatively good dose-response relationship.

[0196] The daily food intake of each administration group began to decrease on the first day after administration and recovered from the second and third days. The daily food intake showed an overall upward trend during administration. On the 28th day after administration, the cumulative food intake of the three dose groups of compound 18# was 58.2g, 46.8g and 36.7g, respectively, which was obviously lower than that of the model control group (70.8g) and showed good dose-dependency. Therefore, compound 18# can obviously reduce the food intake of DIO mice. [Table 16]

[0197] [Table 17]

[0198] Example 11. Improvement effect of some compounds disclosed herein on the glucose metabolism level of db / db mice

[0199] 1. Purpose of the experiment: The improving effect of the numbered compounds on the glucose metabolism level of db / db mice by subcutaneous administration is examined.

[0200] 2. Experimental method: C57BL / KsJ-db / db mice were administered blank vehicle (20 mM sodium citrate + 0.05% Tween-80, pH 7.5), compound LY3298176 (100 nmol / kg body weight) and 18# (three doses, 10 nmol / kg, 30 nmol / kg, 100 nmol / kg body weight, respectively) by subcutaneous injection, and the administration times were days 0, 3, 7, 10, 14, 17, 21, 24 and 27, respectively. Each administration group contained 10 db / db mice. According to the experimental design, tail vein blood was collected by needle on days 0, 7, 14, 21 and 28 to detect fasting blood glucose levels by blood glucose meter and blood glucose test strips, and the mice were fasted for 6 hours before blood was collected at each blood collection time point. On days 3, 10, 17, 24, and 27, blood was collected from the tail vein with a needle and random blood glucose levels were detected using a blood glucose meter. Finally, after the experiment was completed on day 28, animals in all treatment groups were anesthetized with 2% to 5% isoflurane inhalation, and 100 μL of EDTA-K2 anticoagulated whole blood was collected from the orbit and used to measure glycated hemoglobin.

[0201] 3. Experimental results: According to the above experimental method, the specific data are shown in Tables 16 to 18 below.

[0202] [Table 18]

[0203] [Table 19]

[0204] [Table 20]

[0205] 4. Experimental conclusion: In this experiment, at doses of 10 nmol / kg, 30 nmol / kg and 100 nmol / kg, compound 18# of the present disclosure exhibited excellent improving effects on the glucose metabolism level of db / db mice, and showed a significant dose-related relationship. Compound 18# showed a glycated hemoglobin level of 3.78% in the test group at 100 nmol / kg dose at the end of the experiment, while the control compound LY3298176 showed a glycated hemoglobin level of 4.58% in the test group under the same dose conditions, and the efficacy of compound 18# in improving the glucose metabolism level of db / db mice is significantly superior to that of the control compound LY3298176 under the same dose conditions.

[0206] The GLP-1 analog used in Examples 12 to 16 was all compound 18#.

[0207] Example 12. Preparation process of pharmaceutical composition The pharmaceutical composition of the present disclosure can be prepared according to the following process: Step 1: Preparation 1. Preparation of solution 1: Add an appropriate amount of water for injection into a container, and dissolve the prescribed amount of disodium hydrogen phosphate, then add the prescribed amount of GLP-1 analogue or add it last.

[0208] 2. Preparation of solution 2: The prescribed amounts of propylene glycol and phenol were weighed out and added to water for injection in a separate container and stirred until dissolved.

[0209] 3. Solution 2 was added to Solution 1 and stirred uniformly. The pH of the solution was adjusted to 7.4-7.8 with hydrochloric acid or sodium hydroxide solution, and water for injection was added up to the prescribed amount and stirred. Step 2: Sanitization and filtration After preparation was completed, the drug solution was filtered through a sterilization cartridge (pore size: 0.22 μm). The drug solution after sterilization filtration in step 2 was filled into vials at 1.57 mL to 1.67 mL (target filling amount: 1.62 mL), the cap was closed, and visual inspection was performed.

[0210] Example 13. Formulation pH Screening The formulations shown in Table 19 were prepared, and the stability of the formulations was examined at pH 6.5, 7.0, 7.5, 8.0, 8.5 and 9.0 using the properties, related substances and oligopeptides as indicators. The results are shown in Table 20.

[0211] [Table 21]

[0212] [Table 22]

[0213] The results of the stability test showed that the intermediate solution of pH 6.5 was turbid, the related substances were relatively large in the formulations of pH 8.5 and above, and the stability of the formulations of pH 7.0, pH 7.5, and pH 8.0 was relatively good. Considering everything, it was confirmed that the acceptable pH range of the intermediate solution is 7.0 to 8.0.

[0214] Example 14. Buffer concentration screening Using the properties, related substances and oligopeptides as indicators, the stability of the formulations at disodium hydrogen phosphate concentrations of 0 mM, 5 mM, 10 mM and 40 mM was investigated, and the results are shown in Table 21.

[0215] [Table 23]

[0216] The test results showed that the oligopeptide growth was slightly faster in the formulation containing disodium hydrogen phosphate than in the formulation without disodium hydrogen phosphate (0 mM), and that the oligopeptide growth range was larger in the 40 mM formulation, which had a relatively high concentration of disodium hydrogen phosphate, while the oligopeptide growth range was smaller in the 5 mM and 10 mM formulations, and the oligopeptides in each formulation were all within the limit range.

[0217] Example 15. Bacteriostatic agent usage screening GLP-1 analog injections can be used in single dose form or multi-dose package form, and in order to ensure the sterility requirements during use, a bacteriostatic agent should be added in an amount that can suppress the growth of microorganisms in the injection. Phenol with a concentration of 5.5mg / mL was selected, and the relative concentrations of 80%, 100%, and 120% were prepared based on the concentration of 100%, to investigate the bacteriostatic efficacy, and the results are shown in Table 22.

[0218] [Table 24]

[0219] The test results showed that for GLP-1 analogue preparations, when the phenol content was between 4.40 mg / mL and 6.60 mg / mL, all had bacteriostatic effect, and when the phenol concentration was 5.50 mg / mL and 6.60 mg / mL, the bacteriostatic effect was better.

[0220] Example 16. GLP-1 Analog Concentration Screening Using propylene glycol as an osmolality adjuster, samples with GLP-1 analogue concentrations of 2 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 8 mg / mL and 10 mg / mL were prepared, respectively, and the formulations are shown in Table 23.

[0221] [Table 25]

[0222] As can be seen from the data in the above table, (1) All of the samples at each concentration were initially colorless and transparent liquids, indicating good dissolution of the GLP-1 analogues, and no significant differences in pH, related substances, or oligopeptides were observed among the samples at each concentration.

[0223] (2) After being left at 25°C for three months, all samples of each concentration were colorless and transparent liquids, and there was no significant difference in pH value or growth tendency of related substances. The higher the sample concentration, the lower the oligopeptide content. There was no significant difference in impurity growth among the concentration groups, and there was no significant difference in impurity and oligopeptide growth at concentrations of 5mg / mL to 10mg / mL.

[0224] (3) We also found that there was no significant difference in the increase in pH value and related substances when the samples of each concentration were left at 40°C for 10 or 30 days.

[0225] To prepare GLP-1 analog formulations, an attempt was made to replace propylene glycol with the osmotic modifiers glycerin and mannitol, and one exemplary unit dose formulation is shown in Table 24: [Table 26]

[0226] The samples were tested for stability at 40° C. and the stability results are shown in Table 25.

[0227] [Table 27]

[0228] As can be seen from the data in the above table, (1) Initially, each formulation was a colorless, transparent liquid, and the GLP-1 analogues were well dissolved. There were no significant differences in pH, related substances, oligopeptides, or content of each formulation sample.

[0229] (2) Even after leaving each preparation sample at 40°C for 30 days, it remained a transparent liquid, and no granular or sheet-like matter was observed in any of them. Among them, the preparations in group 2, which used mannitol as an osmotic modifier, and the preparation in group 1, which used glycerin as an osmotic modifier, were both pale yellow, but the color change was not very clear. The preparations using glycerin as an osmotic modifier showed a more significant increase in related substances and oligopeptides, and a more significant decrease in the content, compared to the preparations using mannitol as an osmotic modifier.

[0230] When the amount of mannitol used was 4.5% w / v, the osmolality value of the sample was 350 mOsm or more, which is higher than the appropriate osmolality range for the subcutaneous injection route (300±30 mOsm), so the amount of mannitol used was reduced to prepare the sample, and one exemplary unit dose formulation is shown in Table 26. The stability of the sample at 40° C. was examined, and the stability results are shown in Table 27.

[0231] [Table 28]

[0232] [Table 29]

[0233] As can be seen from the data in the above table, (1) Mannitol was used as an osmotic pressure regulator, and the amount of mannitol used was 3.15% w / v and 3.6% w / v. Both were able to completely dissolve the GLP-1 analogue at a concentration of 10 mg / mL, and the dissolution state of the GLP-1 analogue was good. There were no significant differences in the pH, related substances, oligopeptides, or content of each formulation sample.

[0234] (2) After 30 days at 40°C, the formulations containing 3.15% w / v and 3.6% w / v mannitol were pale yellow and transparent, but the color change was not significant and no visible foreign matter was observed. The formulation containing 3.15% w / v mannitol showed a slower growth and decrease in oligopeptide content.

[0235] It will be clearly understood that while the foregoing invention has been described in detail by way of illustration and example, these descriptions and examples should not be construed as limiting the scope of the present disclosure, and the disclosures of all patent and scientific literature cited herein are hereby expressly incorporated by reference in their entirety.

Claims

1. A pharmaceutical composition comprising (a) and (b), (a) a GLP-1 analogue represented by general formula (I) or a medicamentable salt thereof; R １ -X １ -X ２ -Glu-Gly-Thr-Phe-Thr-Ser-Asp-X １０ -Ser-X １２ -X １３ -X １４ -X １５ -X １６ -X １７ -X １８ -X １９ -X ２０ -Glu-Phe-X ２３ -X ２４ -Trp-Leu-X ２７ -X ２８ -X ２９ -X ３０ -Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R ２ (I) Among them, R １ is H, an alkyl group, an acetyl group, a formyl group, a benzoyl group, a trifluoroacetyl group, pGlu, or is absent; R ２ Ha-NH ２ , —OH, or absent; X １ , X ２ , X １０ , X １２ , X １３ , X １４ , X １５ , X １６ , X １７ , X １８ , X １９ , X ２０ , X ２３ , X ２４ , X ２７ , X ２８ , X ２９ and X ３０ are independently selected from any natural or unnatural amino acid residue, and (b) A pharmaceutical composition comprising any one buffer selected from acetate buffers, histidine buffers, phosphate buffers, succinate buffers, and citrate buffers, preferably phosphate buffers, and more preferably disodium hydrogen phosphate.

2. X １ is Tyr or His, and X ２ is Aib or D-Ala, and X １０ is Val or Tyr or Y1, and X １２ is Ser or Ile or Y1, and X １３ is Tyr or Ala or Y1, and X １４ is Leu or Nle or Y1, and X １５ is Asp or Glu, and X １６ is Arg, Glu, Gly, Lys, AiB, or Y1, and X １７ is Glu, Ile or Gln or Y1, and X １８ is Ala, Aib or His, and X １９ is Ala, AiB or Gln, and X ２０ is Gln, Glu, Lys, and X ２３ is Ile or Val, and X ２４ is Ala, Asn or Gln, and X ２７ is Val or Leu, and X ２８ is Arg or Ala, and X ２９ is Gly or Gln, and X ３０ is Gly, Lys, Y1 is a Lys, Orn, Dap, Dab or Cys residue containing a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} ａ -(y-Glu) ｂ -CO-(CH ２ ) ｃ having —COOH, a is an integer from 1 to 3, b is 1 or 2; c is an integer from 10 to 30; The pharmaceutical composition of claim 1.

3. X １ is Tyr, and X ２ is Aib, and X １０ is Tyr, and X １２ is Ile, and X １３ is Tyr, and X １４ is Y1 and X １５ is Asp or Glu, and X １６ is Arg or Lys, and X １７ is Ile, and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln, and X ２３ is Ile or Val, and X ２４ is Asn, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

4. X １６ is Lys, and X ２３ is Val, and X ２７ is Leu, The pharmaceutical composition according to claim 3.

5. X １ is Tyr, and X ２ is Aib or D-Ala, and X １０ is Y1 and X １２ is Ile, and X １３ is Tyr, and X １４ is Leu or Nle, and X １５ is Glu, and X １６ is Arg or Lys, and X １７ is Ile, and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln or Lys, and X ２３ is Ile or Val, and X ２４ is Asn or Gln, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

6. X １ is Tyr, and X ２ is Aib or D-Ala, and X １０ is Tyr, and X １２ is Y1 and X １３ is Tyr, and X １４ is Leu or Nle, and X １５ is Glu, and X １６ is Arg or Lys, and X １７ is Ile, and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln or Lys, and X ２３ is Ile or Val, and X ２４ is Asn or Gln, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

7. X １ is Tyr, and X ２ is Aib or D-Ala, and X １０ is Tyr, and X １２ is Ile, and X １３ is Y1 and X １４ is Leu or Nle, and X １５ is Glu, and X １６ is Arg or Lys, and X １７ is Ile, and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln or Lys, and X ２３ is Ile or Val, and X ２４ is Asn or Gln, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

8. X １ is Tyr, and X ２ is Aib or D-Ala, and X １０ is Tyr, and X １２ is Ile, and X １３ is Tyr, and X １４ is Y1 and X １５ is Glu, and X １６ is Arg or Lys, and X １７ is Ile, and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln or Lys, and X ２３ is Ile or Val, and X ２４ is Asn or Gln, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

9. X ２ is Aib, and X ２０ is Gln, and X ２４ is Asn, The pharmaceutical composition of claim 8.

10. X １ is Tyr, and X ２ is Aib or D-Ala, and X １０ is Tyr, and X １２ is Ile, and X １３ is Tyr, and X １４ is Leu or Nle, and X １５ is Glu, and X １６ is Y1 and X １７ is Ile, and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln or Lys, and X ２３ is Ile or Val, and X ２４ is Asn or Gln, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

11. X ２ is Aib, and X １４ is Leu, and X ２０ is Gln, and X ２４ is Asn, The pharmaceutical composition of claim 10.

12. X １ is Tyr, and X ２ is Aib or D-Ala, and X １０ is Tyr, and X １２ is Ile, and X １３ is Tyr, and X １４ is Leu or Nle, and X １５ is Glu, and X １６ is Arg or Lys, and X １７ is Y1 and X １８ is Ala, and X １９ is Ala, and X ２０ is Gln or Lys, and X ２３ is Ile or Val, and X ２４ is Asn or Gln, and X ２７ is Ile or Leu, and X ２８ is Ala, and X ２９ is Gly, and X ３０ is Gly, Y1 is as defined in claim 2; The pharmaceutical composition of claim 1.

13. a is 2, b is 1 or 2, and c is an integer from 16 to 20; The pharmaceutical composition of claim 2.

14. c is 16, 18 or 20; The pharmaceutical composition of claim 13.

15. Y1 is a Lys residue containing a substituent in the side chain, said substituent being of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} ａ -(y-Glu) ｂ -CO-(CH ２ ) ｃ having —COOH, a is 2, b is 1 or 2; c is 16 or 18; The pharmaceutical composition of claim 2.

16. the substituent is covalently bonded to the amino group on the side chain via an amide bond; A pharmaceutical composition according to any one of claims 2.

17. Y1 is K(—OEG-OEG-yGlu-C18-OH) or K(—OEG-OEG-yGlu-C20-OH), Among them, K(-OEG-OEG-yGlu-C18-OH) has the following structure: 【Chemistry 1】 Preferably it has the following structure: 【Chemistry 2】 K(-OEG-OEG-yGlu-C20-OH) has the following structure: 【Transformation 3】 、 Preferably it has the following structure: The pharmaceutical composition of claim 2. 【Chemistry 4】

18. the substituent is covalently attached to the ε-amino group on the side chain via an amide bond; The pharmaceutical composition of claim 2.

19. The GLP-1 analog has the sequence shown in SEQ ID NO: 20; Preferably, the GLP-1 analogue is selected from the compounds shown in any one of the following numbers 1 to 18: The pharmaceutical composition of claim 1.

20. The GLP-1 analogue is selected from the compounds shown in any one of the following numbers 1# to 18#: The pharmaceutical composition of claim 1.

21. The GLP-1 analog is selected from the compounds shown in Figure 3 as 7#, 12#, 13#, 14#, 15#, 16#, 17#, or 18#; The pharmaceutical composition of claim 1.

22. The pharmaceutical composition further comprises an osmolality adjusting agent; Preferably, the penetration adjuster is one or more selected from propylene glycol, mannitol, sorbitol, xylitol, glycerin, lactose, trehalose, sucrose, glucose, sodium chloride, phosphate, sodium citrate, boric acid, and sodium tartrate; More preferably, the osmolality adjusting agent is propylene glycol, sodium chloride or mannitol; Most preferably, the osmolality adjusting agent is propylene glycol or sodium chloride. A pharmaceutical composition according to any one of claims 1 to 21.

23. The pharmaceutical composition further comprises a bacteriostatic agent; Preferably, the bacteriostatic agent is selected from phenol, o-cresol, m-cresol, p-cresol, methylparaben, propylparaben, 2-phenoxyethanol, butylparaben, 2-phenylethyl alcohol, benzyl alcohol, ethanol, chlorobutanol and thimerosal, bromopropylene glycol, benzoic acid, imidurea, chlorhexidine, sodium dehydroacetate, chlorocresol, ethylparaben, benzethonium chloride or a mixture thereof; More preferably, the bacteriostatic agent is phenol. A pharmaceutical composition according to any one of claims 1 to 21.

24. The pharmaceutical composition further comprises a pH adjuster, and preferably, the pH adjuster is sodium hydroxide and / or hydrochloric acid. A pharmaceutical composition according to any one of claims 1 to 21.

25. the pH of the pharmaceutical composition is 7.0 to 8.0, preferably 7.1 to 7.7, and most preferably about 7.5 or about 7.4; A pharmaceutical composition according to any one of claims 1 to 21.

26. the concentration of the GLP-1 analogue or a pharmaceutically acceptable salt thereof is 0.1 mg / mL to 500 mg / mL, preferably 1.0 mg / mL to 100 mg / mL, more preferably 1.0 mg / mL to 30.0 mg / mL, and most preferably 2.0 mg / mL to 10.0 mg / mL; A pharmaceutical composition according to any one of claims 1 to 21.

27. The concentration of the buffer in the pharmaceutical composition is 1.0 mM to 35.0 mM, preferably 1.0 mM to 25.0 mM, more preferably 2.0 mM to 10.0 mM. A pharmaceutical composition according to any one of claims 1 to 21.

28. The concentration of propylene glycol in the pharmaceutical composition is 10 mg / mL to 20 mg / mL, preferably 12 mg / mL to 16 mg / mL, more preferably about 14 mg / mL; alternatively, the concentration of mannitol in the pharmaceutical composition is 30 mg / mL to 45 mg / mL, preferably 30 mg / mL to 40 mg / mL, more preferably about 31.5 mg / mL; alternatively, the concentration of sodium chloride in the pharmaceutical composition is 2 mg / mL to 18 mg / mL, preferably 8 mg / mL to 10 mg / mL, more preferably about 9 mg / mL; 23. The pharmaceutical composition of claim 22.

29. The concentration of the bacteriostatic agent in the pharmaceutical composition is 4.0 mg / mL to 7.0 mg / mL, preferably 4.4 mg / mL to 6.8 mg / mL, more preferably 5.5 mg / mL to 6.6 mg / mL, and most preferably about 5.5 mg / mL; 24. The pharmaceutical composition of claim 23.

30. 1. A pharmaceutical composition comprising: The compound shown in 18# of Figure 3 or a medicinal salt thereof; Sodium dihydrogen phosphate, and containing propylene glycol, mannitol or sodium chloride, Optionally, the pharmaceutical composition further comprises phenol. Pharmaceutical compositions.

31. A pharmaceutical composition comprising any one of A) to J), and the pH of the pharmaceutical composition is 6.5 to 9.0, wherein: A) 1.0 mg / mL to 100 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate, and 10 mg / mL to 20 mg / mL propylene glycol or 30 mg / mL to 40 mg / mL mannitol; Or, B) 1.0 mg / mL to 100 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate, and containing 2 mg / mL to 18 mg / mL sodium chloride; Or, C) 1.0 mg / mL to 100 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate; comprising 10 mg / mL to 20 mg / mL propylene glycol, or 15 mg / mL to 45 mg / mL mannitol, or 2 mg / mL to 18 mg / mL sodium chloride; Optionally, the pharmaceutical composition further comprises 4.0 mg / mL to 7.0 mg / mL of an antibacterial agent, such as phenol; D) 1.0 mg / mL to 30.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 1.0 mM to 25.0 mM sodium dihydrogen phosphate; 11 mg / mL to 18 mg / mL propylene glycol, or 20 mg / mL to 40 mg / mL mannitol, or 3 mg / mL to 15 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.2 mg / mL to 6.9 mg / mL; and the pH of the pharmaceutical composition is 7.0 to 8.0; E) 2.0 mg / mL to 10.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 2.0 mM to 10.0 mM sodium dihydrogen phosphate; 12 mg / mL to 16 mg / mL propylene glycol, or 25 mg / mL to 35 mg / mL mannitol, or 8 mg / mL to 10 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.4 mg / mL to 6.8 mg / mL; and the pH of the pharmaceutical composition is 7.1 to 7.7; F) 1.0 mg / mL to 100 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 1.0 mM to 35.0 mM sodium dihydrogen phosphate; 10 mg / mL to 20 mg / mL propylene glycol, or 15 mg / mL to 45 mg / mL mannitol, or 2 mg / mL to 18 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.0 mg / mL to 7.0 mg / mL; and water for injection, and the pH of the pharmaceutical composition is 6.5 to 9.0; G) 1.0 mg / mL to 30.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 1.0 mM to 25.0 mM sodium dihydrogen phosphate; 11 mg / mL to 18 mg / mL propylene glycol, or 20 mg / mL to 40 mg / mL mannitol, or 3 mg / mL to 15 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.2 mg / mL to 6.9 mg / mL; and water for injection, and the pH of the pharmaceutical composition is 7.0 to 8.0; H) 2.0 mg / mL to 20.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 2.0 mM to 10.0 mM sodium dihydrogen phosphate; 12 mg / mL to 16 mg / mL propylene glycol, or 25 mg / mL to 35 mg / mL mannitol, or 7 mg / mL to 10 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.4 mg / mL to 6.8 mg / mL; and water for injection, and the pH of the pharmaceutical composition is 7.1 to 7.7; I) 5.0 mg / mL to 15.0 mg / mL or 5.0 mg / mL to 10.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 5.0 mM to 10.0 mM sodium dihydrogen phosphate; 12 mg / mL to 16 mg / mL propylene glycol, or 7 mg / mL to 10 mg / mL sodium chloride, or 8 mg / mL to 9 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.4 mg / mL to 6.8 mg / mL; and water for injection, and the pH of the pharmaceutical composition is 7.1 to 7.7; J) 5.0 mg / mL to 15.0 mg / mL or 5.0 mg / mL to 10.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof; 4.0 mM to 8.0 mM or 4.0 mM to 6.0 mM sodium dihydrogen phosphate; 12 mg / mL to 16 mg / mL propylene glycol, or 7 mg / mL to 10 mg / mL sodium chloride, or 8 mg / mL to 9 mg / mL sodium chloride; an optional antimicrobial agent such as phenol at 4.4 mg / mL to 6.8 mg / mL; and water for injection, and the pH of the pharmaceutical composition is 7.1 to 7.

7.

31. The pharmaceutical composition of claim 30.

32. (1) The pharmaceutical composition comprises about 2.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL of propylene glycol or about 31.5 mg / mL of mannitol, and about 5.5 mg / mL of phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.4; (2) The pharmaceutical composition comprises about 4.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL propylene glycol or about 31.5 mg / mL mannitol, and about 5.5 mg / mL phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.4; (3) The pharmaceutical composition comprises about 5.0 mg / mL of the compound shown in 18# of FIG. 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL propylene glycol or about 31.5 mg / mL mannitol, and about 5.5 mg / mL phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.4; (4) The pharmaceutical composition comprises about 6.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL propylene glycol or about 31.5 mg / mL mannitol, and about 5.5 mg / mL phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.4; (5) The pharmaceutical composition comprises about 8.0 mg / mL of the compound shown in 18# of FIG. 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL propylene glycol or about 31.5 mg / mL mannitol, and about 5.5 mg / mL phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.4; (6) The pharmaceutical composition comprises about 10.0 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 14.0 mg / mL propylene glycol or about 31.5 mg / mL mannitol, and about 5.5 mg / mL phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.4; or (7) The pharmaceutical composition comprises about 2.0 mg / mL, about 4.0 mg / mL, about 5.0 mg / mL, about 6.0 mg / mL, about 8.0 mg / mL, 10 mg / mL, or about 20 mg / mL of the compound shown in 18# of Figure 3 or a pharmaceutically acceptable salt thereof, about 5.0 mM sodium dihydrogen phosphate, about 9 mg / mL sodium chloride, and optionally about 5.5 mg / mL phenol, and the pH of the pharmaceutical composition is about 7.5 or about 7.

4. The pharmaceutical composition according to any one of claims 30 to 31.

33. 10. A method for preparing the pharmaceutical composition of claim 1, comprising: The method comprises the step of dissolving said GLP-1 analogue or a pharmaceutically acceptable salt thereof.

34. A lyophilized formulation comprising:

10. A freeze-dried formulation, wherein the freeze-dried formulation can be reconstituted to form the pharmaceutical composition of claim 1, or the freeze-dried formulation is obtained by freeze-drying the pharmaceutical composition of claim 1.

35. 1. A method for producing a reconstituted solution, comprising: A method comprising the step of reconstituting the lyophilized formulation of claim 17.

36. A product, 35. An article of manufacture comprising the pharmaceutical composition of claim 1 or the lyophilized formulation of claim 34.

37. Use of the pharmaceutical composition of claim 1, the lyophilized formulation of claim 34, or the product of claim 36 in the preparation of a medicament for treating non-insulin-dependent diabetes mellitus, insulin-dependent diabetes mellitus, obesity, non-alcoholic fatty liver disease, hepatic steatosis, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, insulin resistance, dyslipidemia associated with insulin resistance, and / or dyslipidemia associated with diabetes.

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