GLP-1 / GIP dual agonist as well as preparation method and application thereof

Abstract

The invention relates to a long-acting GLP-1 / GIP dual agonist compound, the compound has dual agonist effects on a glucagon-like peptide-1 (GLP-1) receptor and a human glucose-dependent insulinotropic polypeptide (GIP) receptor, and the invention also relates to application of the compound in preparation of a medicine for treating diabetes mellitus.

Description

GLP-1 / GIP dual agonist and its preparation method and use Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a GLP-1 / GIP dual agonist and its use in preparing a medicine for treating diabetes. Background Art

[0002] Type 2 diabetes is a chronic metabolic disease characterized by hyperglycemia due to insulin resistance. Elevated blood sugar levels can increase the incidence and mortality of various other diseases. Current treatments, such as increased exercise, dietary control, oral hypoglycemic medications, and injectable medications, can only temporarily maintain blood sugar balance but are unable to fundamentally treat diabetes and the many complex complications caused by hyperglycemia. Therefore, the active exploration and development of novel, safe, and effective diabetes treatments is of great clinical significance.

[0003] Glucagon-like peptide-1 (GLP-1) is a hormone primarily produced by intestinal L cells and belongs to the incretin class. GLP-1 is an important incretin hormone secreted by intestinal L cells. Its primary physiological effects in the body include stimulating insulin secretion and release, inhibiting glucagon secretion, promoting pancreatic β-cell proliferation and inhibiting apoptosis, inhibiting gastric emptying, and promoting satiety. Glucose-dependent insulinotropic polypeptide (GIP), which regulates blood sugar through insulinotropic and glucagon-stimulating effects, has also been widely demonstrated and applied.

[0004] Compounds with GLP-1 / GIP dual agonist activity have been described in WO2013164483A1, WO2014192284A1, and WO2016111971A1.

[0005] Fatty acid modification of peptides can, for example, extend the half-life of peptides by physically binding their fatty acid chains to albumin subdomain motifs, thereby improving the pharmacokinetics of peptides. Although the use of fatty acids can improve the half-life of peptides, the specific effect of the extension and some side effects remain technical issues that need to be addressed.

[0006] A series of side effects regarding GLP-1 include: for example, because animal studies and analysis of drug databases have shown that GLP-1 receptor agonists are associated with pancreatitis, pancreatic cancer, and thyroid cancer, there are concerns that these drugs will affect pancreatic and thyroid tissue. Other case reports have linked the use of GLP-1 (mainly exenatide) to the occurrence of acute kidney injury, mainly due to hemodynamic disorders caused by nausea, vomiting, and diarrhea. The most common symptoms associated with the use of GLP-1 receptor agonists are gastrointestinal symptoms, mainly nausea. Other common adverse reactions include injection site reactions, headaches, and nasopharyngitis. (Filippatos TD, Panagiotopoulou TV, Elisaf MS. Adverse Effects of GLP-1 Receptor Agonists. Rev Diabet Stud. 2014 Fall-Winter; 11(3-4):202-30.doi:10.1900 / RDS.2014.11.202.Epub 2015 Feb 10.PMID:26177483; PMCID:PMC5397288.)

[0007] Regarding the side effects of tirzepatide (Mathiesen DS, Bagger JI, Bergmann NC, Lund A, Christensen MB, T, Knop FK. The Effects of Dual GLP-1 / GIP Receptor Agonism on Glucagon Secretion-A Review. Int J Mol Sci. 2019 Aug 22;20(17):4092. doi:10.3390 / ijms20174092. PMID:31443356; PMCID:PMC6747202.) At the highest dose, tirzepatide exceeded dulaglutide in the number of adverse events, particularly gastrointestinal adverse events (66.0% of patients taking 15 mg tirzepatide and 42.6% of patients taking 1.5 mg dulaglutide), and also exceeded dulaglutide in the total number of hypoglycemic episodes (7.5% of participants taking 15 mg tirzepatide and 3.7% of participants taking dulaglutide) (Frias JP, Nauck MA, Van J., Kutner ME, Cui X., Benson C., Urva S., Gimeno RE, Milicevic Z., Robins D., et al. Efficacy and safety of LY3298176, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes: a randomized, placebo-controlled and active comparator-controlled phase 2 trial.Lancet.2018;392:2180–2193.doi:10.1016 / S0140-6736(18)32260-8.).To circumvent this problem, a titration strategy to reduce adverse events of tirzepatide has been reported (Frias JP, Nauck MA, Van J., Benson C., Bray R., Milicevic Z., Haupt A., Robins DA993-P: A 12-week, randomized, placebo-controlled study assessing the efficacy and safety of three dose-escalation algorithms of tirzepatide, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes. Diabetes. 2019; 68: 993. doi: 10.2337 / db19-993-P.). However, compared with previously reported studies, a three-step dose-escalation regimen of 15 mg tirzepatide did not appear to be effective in reducing the overall incidence of gastrointestinal side effects (Frias JP, Nauck MA, Van J., Kutner ME, Cui X., Benson C., Urva S., Gimeno RE, Milicevic Z., Robins D., et al. Efficacy and safety of LY3298176, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes: a randomised, placebo-controlled and active comparator-controlled phase 2 trial. Lancet. 2018; 392: 2180–2193. doi: 10.1016 / S0140-6736(18)32260-8.). The incidence of hypoglycemic episodes also increased to 16% in participants taking 15 mg of tilbotide. Although tilbotide treatment was still associated with promising weight loss (approximately 5.6 kg after 12 weeks), the high incidence of gastrointestinal side effects does cast some uncertainty on the clinical potential of tilbotide. In addition, the titration strategy in the marketing instructions for tilbotide takes a long time. The starting dose is 2.5 mg, which is increased to 5 mg after 4 weeks. If the dose needs to be increased to control blood sugar, the current dose must be maintained for 4 weeks and then increased by 2.5 mg, which is very inconvenient.

[0008] There is still a medical need to provide new compounds that are dual agonists of GIP and GLP-1 receptors and support the possibility of once-weekly dosing in humans, providing options with better safety, better efficacy and more convenient medication.

[0009] Summary of the Invention

[0010] One object of the present invention is to provide certain long-acting GLP-1 / GIP dual agonist compounds. Compared with GIP / GLP-1 dual agonist compounds in the art, the compounds of the present invention have a longer half-life and / or a more sustained duration of action. The compounds of the present invention have a stronger hypoglycemic effect. The compounds of the present invention have lower side effects, including but not limited to gastrointestinal reactions and cardiac safety risks. The compounds of the present invention can adopt a shorter titration strategy. The compounds of the present invention have a larger safety window. In addition, the compounds of the present invention have a protective effect on the liver.

[0011] As part of the present invention, applicants have discovered that the selection of the length, composition and attachment position of the fatty acid chain, as well as the linker between the peptide and the fatty acid chain, has an unexpected effect on extending the half-life of the polypeptide.

[0012] One embodiment of the present invention is to provide a compound of formula (AI) and a pharmaceutically acceptable salt thereof:

[0013] Y-X1-EGT-X2-TSDY-A11-A12-A13-LDK-A17-AQ-A20-EFVKWLLK-A29-GPSSGAPPPSK (AI);

[0014] wherein X1 is Aib; X2 is αMePhe;

[0015] A11 is Aib or Ala;

[0016] A12 is Ala, Ile, Lys, Phe, or Pya (4);

[0017] A13 is Aib, Cha, Leu, αMePhe, or αMeTyr;

[0018] A17 is Gln or Ile;

[0019] A20 is Ala or Ser;

[0020] A29 is Gln or Gly;

[0021] 1 K, 2 K, 3 K or 4 K positions are selected from the 16th, 24th, 28th and 40th positions by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl)a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by being conjugated to the ε-amino group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 24, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester.

[0022] In another embodiment, the present invention provides a compound of formula (AI) or a pharmaceutically acceptable salt thereof, wherein,

[0023] X1 is Aib; X2 is αMePhe;

[0024] A11 is Aib or Ala;

[0025] A12 is Ala, Ile, Lys, Phe, or Pya (4);

[0026] A13 is Aib, Cha, Leu, αMePhe, or αMeTyr;

[0027] A17 is Gln or Ile;

[0028] A20 is Ala or Ser;

[0029] A29 is Gln or Gly,

[0030] 1 K, 2 K, 3 K or 4 K positions are selected from the 16th, 24th, 28th and 40th positions by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by being conjugated to the ε-amino group of the K side chain, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester; and compounds wherein c is independently selected from 14, 16, 18 or 20 are also provided.

[0031] The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A11 is Aib; or / and

[0032] The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A12 is Ile; or / and

[0033] The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A13 is Aib; or / and

[0034] The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A17 is Gln; or / and

[0035] The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A20 is Ala; or / and

[0036] The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A29 is Gly.

[0037] One embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0038] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0039] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; 1, 2, 3 or 4 K positions are selected from positions 16, 24, 28 and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugation to the ε-amino group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 24, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0040] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein one K position is selected from among positions 16, 24, 28, and 40 by replacing the K with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugating to the ε-amino group of the K side chain. For example, at the position of K at position 16, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugating to the ε-amino group of the K side chain; for example, at the 24-position of K, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a-(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugating to the ε-amino group of the K side chain; for example, at position 28 of K, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugating to the ε-amino group of the K side chain; for example, at position 40 of K, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugating to the ε-amino group of the K side chain.

[0041] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the positions of 2 K are selected from among the K at positions 16, 24, 28, and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains. For example, at the positions of K at position 16 and K at position 24, ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to carry out chemical modification of two modified chains; for example, at the positions of K at position 16 and K at position 28, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains; for example, at the positions of K at position 16 and K at position 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains; for example, at the positions of K at position 24 and K at position 28, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c-Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains; for example, at the positions of K at position 24 and K at position 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains; for example, at the positions of K at position 28 and K at position 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains.

[0042] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the positions of 3 K are selected from among the K at positions 16, 24, 28, and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of three modified chains. For example, at the positions of K at position 16, K at position 24, and K at position 28, ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to carry out chemical modification of three modified chains; for example, at the positions of K at 16, K at 24, and K at 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of three modified chains; for example, at the positions of K at position 16, K at position 28, and K at position 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of three modified chains; for example, at the positions of K at position 24, K at position 28, and K at position 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of three modified chains.

[0043] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the positions of 4 K are selected from among positions 16, 24, 28, and 40 by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is conjugated to the ε-amino group of the K side chain to perform chemical modification of four modified chains. For example, at the positions of K 16, K 24, K 28 and K 40, ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z was conjugated to the ε-amino group of the K side chain to perform chemical modification of the four modified chains.

[0044] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester, for example, but not limited to, the Z may include carboxylic acid (-CO2H) or carboxylic acid bioisostere (e.g. ), phosphonic acid (-P(O)(OH)2) or sulfonic acid (-SO2OH) groups, preferably -CO2H.

[0045] Carboxylic acid bioisosteres, suitable carboxylic acid bioisosteres are known in the art. Preferably, the bioisostere has a pK a Examples of suitable bioisosteres may include, but are not limited to, tetrazoles, acylsulfonamides, acylhydroxylamines, and squaric acid derivatives, as shown below:

[0046] R is Me or CF3.

[0047] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, preferably 1, 2 or 3, b is independently selected from an integer of 1, 2, 3, 4 or 5, preferably 1, 2 or 3, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0048] One embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0049] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0050] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; one or two K positions are selected from positions 16, 24, 28, and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugation to the ε-amino group of the K side chain, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester, preferably -CO2H, each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 24; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0051] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; one or two K positions are selected from positions 16, 24, 28, and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by being conjugated to the ε-amino group of the K side chain, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester, preferably -CO2H, a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0052] One embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0053] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0054] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain and chemically modified, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

[0055] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is chemically modified by conjugating to the ε-amino group of the K side chain, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0056] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0057] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0058] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain and chemically modified, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

[0059] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is chemically modified by conjugating to the ε-amino group of the K side chain, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0060] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0061] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0062] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 16 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain and chemically modified, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

[0063] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 16 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is chemically modified by conjugating to the ε-amino group of the K side chain, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0064] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0065] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0066] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain and chemically modified, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

[0067] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is chemically modified by conjugating to the ε-amino group of the K side chain, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0068] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0069] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0070] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 24 and 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c-CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0071] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 24 and 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0072] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0073] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0074] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 16 and 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0075] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 16 and 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2)c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0076] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0077] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0078] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 16 and 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0079] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 16 and 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0080] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0081] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0082] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 16 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0083] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 16 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0084] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0085] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0086] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 24 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c-CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0087] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 24 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0088] Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof:

[0089] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I);

[0090] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 28 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

[0091] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at positions 28 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c-CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of two modified chains, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

[0092] In all embodiments described above, each a is independently an integer from 1 to 3, each b is independently an integer from 1 to 3, and each c is independently an integer from 12 to 22; in addition, the present invention provides compounds wherein c is 14, 16, 18 or 20. In all embodiments described above, preferred embodiments are those in which a modified chain is modified at K at position 24 or 28.

[0093] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0094] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0095] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 7).

[0096] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0097] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0098] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 18 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 13).

[0099] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0100] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0101] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 14).

[0102] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0103] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0104] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)3-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 15).

[0105] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0106] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0107] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 20The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 16).

[0108] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0109] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0110] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 8).

[0111] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0112] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0113] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 17).

[0114] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0115] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0116] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)3-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 18).

[0117] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0118] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0119] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 19).

[0120] In one embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof;

[0121] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0122] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 20).

[0123] The present invention also provides a method for increasing the duration of action of a GLP-1 / GIP dual agonist in a patient, characterized by using the acylated GLP-1 / GIP dual agonist provided by the present invention as described above.

[0124] In one embodiment, the present invention provides a composition comprising a compound of the present invention and a pharmaceutically acceptable carrier, diluent, or excipient.

[0125] One embodiment provides the use of a compound according to any one of the above embodiments in the preparation of a medicament for treating or preventing hyperglycemia, impaired glucose tolerance, type I diabetes, type II diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular diseases, stroke, inflammatory bowel syndrome, dyspepsia, and gastric ulcer.

[0126] One embodiment provides the use of a compound according to any one of the above embodiments for the preparation of a medicament for delaying or preventing disease progression in type II diabetes.

[0127] One embodiment provides the use of a compound according to any one of the above embodiments for the preparation of a medicament for reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell mass and / or restoring glucose sensitivity of β-cells.

[0128] In one embodiment, the present invention provides a method of treating type II diabetes comprising administering to a patient in need thereof an effective amount of a compound of the present invention.

[0129] In one embodiment, the present invention provides a method for improving glycemic control in a patient with type 2 diabetes, comprising administering to a patient in need thereof an effective amount of a compound of the present invention as an adjunct to diet and exercise.

[0130] In one embodiment, the present invention provides a method for long-term weight management in a patient with an initial body mass index ≥ 27 and type 2 diabetes, comprising administering to a patient in need thereof an effective amount of a compound of the present invention as an adjunct to a reduced-calorie diet and increased physical activity.

[0131] In one embodiment, the present invention provides a method for treating metabolic syndrome, comprising administering to a patient in need thereof an effective amount of a compound of the present invention. In another embodiment, the present invention provides a method for treating dyslipidemia, obesity, and / or hepatic steatosis associated with insulin resistance and diabetes, comprising administering to a patient in need thereof an effective amount of a compound of the present invention. Additionally, the present invention provides a method for treating frailty or increasing bone strength, comprising administering to a patient in need thereof an effective amount of a compound of the present invention.

[0132] In one embodiment, the present invention provides compounds of the present invention for use as an adjunct to diet and exercise for glycemic control in patients with type 2 diabetes. In one embodiment, the present invention provides compounds of the present invention for use as an adjunct to a hypocaloric diet and increased physical activity for long-term weight management in patients with an initial body mass index ≥ 27 and type 2 diabetes.

[0133] The compounds of the present invention can be used as drugs for treating or preventing various diseases, including obesity, due to the aforementioned activation of GLP-1 receptors and GIP receptors. The compounds of the present invention can be used as drugs for treating or preventing various diseases, including obesity. The compounds of the present invention can be used as drugs for treating or preventing the following diseases: for example, symptomatic obesity, obesity based on simple obesity, disease states or diseases related to obesity, eating disorders, diabetes (for example, type I diabetes, type II diabetes, gestational diabetes, obesity-related diabetes), hyperlipidemia (for example, hypertriglyceridemia, hypercholesterolemia, high LDL-cholesterolemia, low HDL-cholesterolemia, postprandial hyperlipidemia), hypertension, heart failure, complications of diabetes (for example, neuropathy, nephropathy, retinopathy, diabetic cardiomyopathy, cataracts, macroangiopathy, osteopenia, hyperosmolar diabetic coma), infectious diseases (for example, respiratory tract infection, urinary tract infection, gastrointestinal infection, superficial and soft tissue infection, lower limb infection), Diabetic gangrene, xerostomia, hearing loss, cerebrovascular disorders, peripheral blood circulation disorders, metabolic syndrome (a disease state having three or more selected from hypertriglyceridemia (TG), low HDL cholesterolemia (HDL-C), hypertension, abdominal obesity and impaired glucose tolerance), sarcopenia, etc.

[0134] Examples of symptomatic obesity include endocrine obesity (e.g., Cushing's syndrome, hypothyroidism, insulinoma, obese type II diabetes, pseudohypoparathyroidism, hypogonadism), central obesity (e.g., hypothalamic obesity, frontal lobe syndrome, Creutzfeldt-Lewis syndrome), genetic obesity (e.g., Prader-Willi syndrome, Lawrence-Moore-Biedl syndrome), drug-induced obesity (e.g., obesity induced by steroids, phenothiazines, insulin, sulfonylurea (SU) drugs, beta-blockers), and the like.

[0135] Examples of disease states or diseases associated with obesity include: glucose tolerance disorders, diabetes (especially type II diabetes, obese diabetes), lipid metabolism disorders (synonymous with the above-mentioned hyperlipidemia), hypertension, heart failure, hyperuricemia, fatty liver (including non-alcoholic hepatitis), coronary heart disease (myocardial infarction, angina pectoris), cerebral infarction (cerebral thrombosis, transient ischemic attack), bone / joint diseases (knee osteoarthritis, hip osteoarthritis, ankylosing spondylitis, low back pain), sleep apnea syndrome / Pickwick syndrome, menstrual disorders (abnormal menstrual cycle, amenorrhea, abnormal menstrual symptoms), metabolic syndrome, etc.

[0136] The compound provided by the present invention has GLP-1 / GIP dual agonist activity, a long half-life, and shows a longer duration of drug effect.

[0137] The compounds of the present invention can react with any of a variety of inorganic and organic acids to form pharmaceutically acceptable acid addition salts. Pharmaceutically acceptable salts and common methods for preparing them are well known in the art.

[0138] The amino acid sequences of the present invention contain the standard one-letter or three-letter codes for the twenty naturally occurring amino acids. Additionally, "Aib" is α-aminoisobutyric acid, "αMePhe" is α-methylphenylalanine, "Pya(4)" is 4-pyridylalanine, "Cha" is cyclohexylalanine, and "αMeTyr" is α-methyltyrosine.

[0139] The "AEEA" of the present invention is the abbreviation of 2-(2-amino-ethoxy)-ethoxy]-acetyl. The "Ste" of the present invention represents an octadecane dioic acid-C(O)-C 16 H 32 -C(O)-.

[0140] The "effective amount" or "therapeutically effective amount" of the present invention is the amount or dosage of a compound of the present invention or a pharmaceutically acceptable salt thereof, which, when administered to a patient in single or multiple doses, provides the desired effect in the patient being diagnosed or treated.

[0141] The "effective dose" as used herein refers to the minimum dose that produces a pharmacological effect. Specifically, in the db / db mouse multiple-dose efficacy study, it refers to the minimum dose that produces a significant difference (P < 0.05) in blood glucose AUC in the test group compared to the vehicle group.

[0142] The term "efficacy dose" as used herein refers to the dose that produces a pharmacologically effective effect. Specifically, in the db / db mouse multiple-dose efficacy study, it refers to the dose at which the blood glucose AUC in the test group shows a significant difference (P < 0.05) compared to the vehicle group.

[0143] The "Vehicle group" described in the present invention refers to the vehicle control group.

[0144] The "AUC" in this invention refers to the area under the drug-time curve, which is the area enclosed by the drug concentration curve over the time axis. This parameter is an important indicator for evaluating the degree of drug absorption and reflects the drug's exposure characteristics in the body.

[0145] The "MRT" mentioned in the present invention refers to the mean residence time, which is the average time that drug molecules stay in the body, and represents the time required to eliminate 63.2% of the drug from the body.

[0146] The "MTD" mentioned in the present invention refers to the maximum tolerated dose, which is the highest dose that does not cause the death of the test animals.

[0147] The "internal standard" mentioned in the present invention refers to a known amount of a pure compound added to a sample to correct errors caused by instrument signal fluctuations, human operation, etc.

[0148] The "blood glucose AUC inhibition rate" of the present invention is the percentage of blood glucose AUC reduction in the test substance administration group compared with the vehicle group. Blood glucose AUC inhibition rate = (blood glucose AUC Vehicle - Blood glucose AUC 受试物 ) / blood glucose AUC Vehicle

[0149] The "titration strategy" described in the present invention is a method of increasing the dose, that is, after the initial dose, the frequency of the dose increase and the amount of administration are adjusted to obtain the optimal dosage.

[0150] The "patient" described in the present invention includes humans or animals, preferably humans.

[0151] The "modified chain" of the present invention is ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c-Z, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; further wherein a is independently selected from an integer from 1, 2, 3, 4 or 5, b is independently selected from an integer from 1, 2, 3, 4 or 5, and c is independently selected from an integer from 12 to 22; in addition, the present invention provides compounds wherein c is 14, 16, 18 or 20, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester, for example, but not limited to, the Z may include carboxylic acid (-CO2H) or carboxylic acid bioisostere (e.g. ), phosphonic acid (-P(O)(OH)2) or sulfonic acid (-SO2OH) groups, preferably -CO2H.

[0152] Carboxylic acid bioisosteres, suitable carboxylic acid bioisosteres are known in the art. Preferably, the bioisostere has a pK a Examples of suitable bioisosteres may include, but are not limited to, tetrazoles, acylsulfonamides, acylhydroxylamines, and squaric acid derivatives, as shown below:

[0153] R is Me or CF3.

[0154] For the peptides mentioned herein, according to conventional peptide notation, the left end is the N-terminus (amino terminus) and the right end is the C-terminus (carboxyl terminus). The C-terminus of the peptide can be an amide (-CONH2), a carboxyl (-COOH), a carboxylate (-COO - ), any of alkylamides (-CONHR') and esters (-COOR'), R' is C 1-8 An alkyl group. In particular, an amide (-CONH2) is preferred.

[0155] The "coupling" or "accessioning" in the preparation method of the present invention refers to the process of adding new amino acids to the bound amino acids or peptides. Description of the drawings:

[0156] Figures 1A-1B show blood glucose changes in db / db mice after drug administration. Figure 1A shows blood glucose changes in db / db mice after a single subcutaneous administration of 10 nmol / kg of P007, P008, P014, P019, Tirzepatide, and P001; Figure 1B shows blood glucose changes in db / db mice after a single subcutaneous administration of 10 nmol / kg of P013, P015, P016, P017, P018, P020, and Tirzepatide.

[0157] Figures 2A-2C show the blood glucose AUC after dosing in db / db mice. Figure 2A shows the AUC of P007, P008, P014, P019, Tirzepatide, and P001 at 0-48 hours after a single subcutaneous dose of 10 nmol / kg; Figure 2B shows the AUC of P007, P008, P014, P019, and Tirzepatide at 0-72 hours after a single subcutaneous dose of 10 nmol / kg; Figure 2C shows the AUC of P013, P015, P016, P017, P018, P020, and Tirzepatide at 0-56 hours after a single subcutaneous dose of 10 nmol / kg. In Figure 2A, T-test, *p<0.05, **p<0.01, ***p<0.001 compared with Vehicle; §p<0.05, §§p<0.01, §§§p<0.001 compared with P001. In Figure 2B, T-test, *p<0.05, **p<0.01, ***p<0.001 compared with Vehicle; #p<0.05, ##p<0.01 compared with Tirzepatide. In Figure 2C, T-test, *p<0.05, **p<0.01, ***p<0.001 compared with Vehicle; #p<0.05 compared with Tirzepatide.

[0158] Figures 3A-3D show changes in blood glucose levels in db / db mice after multiple dosing. Figure 3A compares blood glucose levels in db / db mice after multiple subcutaneous administration of P014 and Tirzepatide; Figure 3B compares blood glucose levels in db / db mice after multiple subcutaneous administration of P016 and Tirzepatide; Figure 3C compares blood glucose levels in db / db mice after multiple subcutaneous administration of P017 and Tirzepatide; and Figure 3D compares blood glucose levels in db / db mice after multiple subcutaneous administration of P020 and Tirzepatide.

[0159] The present invention also includes novel intermediates and methods that can be used to synthesize the compounds of the present invention or their pharmaceutically acceptable salts. The intermediates and compounds of the present invention can be prepared by a variety of methods known in the art. In particular, methods using chemical synthesis are illustrated in the following examples. The specific synthetic steps of each described approach can be combined in different ways to prepare the compounds of the present invention or their salts. Reagents and raw materials are readily available to those of ordinary skill in the art. It should be understood that these examples are not intended to limit the scope of the present invention in any way. Mass spectrometry was performed using an Agilent 1260 / 6110 liquid chromatograph-mass spectrometer with a scan range of 100-1500.

[0160] Example 1:

[0161] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0162] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)3-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 15).

[0163] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K24, for which the structure has been unfolded.

[0164] The preparation method adopts the Fmoc solid phase peptide synthesis strategy, and the method includes:

[0165] (1) Synthesis of peptide resin intermediate 1

[0166] Take Fmoc-Rink Linker-Nle-MBHA resin (S = 0.49 mmol / g), swell it with an appropriate amount of DCM, and wash it with DCM 2-3 times. Deprotect it with 20% PIP / DMF solution for 30 minutes, filter and wash it to obtain the Fmoc-free NH2-Rink linker-Nle-MBHA resin, and drain the solvent for use.

[0167] Take 4 equivalents of Fmoc-Lys(Boc)-OH and HOBt respectively, dissolve them in appropriate amount of DMF / DCM; take another 4 equivalents of DIC, dilute it by half with DCM, slowly add it to the DMF / DMC solution with stirring, and stir the reaction at -5~0℃ for not less than 60 minutes. After activation, set aside.

[0168] Add the activated Fmoc-Lys(Boc)-OH solution to the NH2-Rink linker–Nle-MBHA resin, control the reaction temperature at 10-30°C, and conduct the coupling reaction for 240–480 min. Filter and wash to obtain the Fmoc-Lys(Boc)-Rink linker–Nle-MBHA resin. Deprotect the resin with a 20% PIP / DMF solution for 30 min, filter and wash to obtain the de-Fmoc Lys(Boc)-Rink linker–Nle-MBHA resin.

[0169] Under the same reaction conditions, start from the second amino acid at the C-terminus and couple to the N-terminus one by one. If the coupling is incomplete (color reaction), use HBTU / DIEA for a second condensation to ensure that each amino acid is completely condensed. The coupling sequence is Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH.H2O, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly- Gly-OH, Fmoc-Lys(Boc)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH.H2O, Fmoc-Ala-OH.H2O , Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH.H2O, Fmoc-Gln(Trt)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Aib-OH, Fmoc-Ile-OH, Fmoc-Aib-OH, Fmoc-Tyr( tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-α-Me-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Glu(OtBu)-Gly-OH, Fmoc-Aib-OH, Boc-Tyr(tBu)-OH.

[0170] get:

[0171] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-A ib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Mtt)-Trp(Boc)-Leu-Le u-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin

[0172] After the above condensation is complete, the Mtt protection is removed with 50% HFIP / DCM solution for 30 minutes, followed by washing and filtration to obtain the peptide resin intermediate 1:

[0173] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala- Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin.

[0174] (2) Modification chain modification step

[0175] Dissolve 4 equivalents of Fmoc-AEEA-OH and HOBt in an appropriate amount of DMF / DCM. Dissolve another 4 equivalents of DIC in DCM by half, then slowly add it to the DMF / DCM solution with stirring. Stir and react at -5 to 0°C for at least 60 minutes. Activate and set aside.

[0176] The activated Fmoc-AEEA-OH solution was added to the previously swollen and washed peptide resin intermediate 1. The reaction temperature was controlled at 10-30°C. The coupling reaction was carried out for 240-480 minutes. The mixture was filtered and washed. The Fmoc protection was removed with a 20% PIP / DMF solution for 30 minutes. The mixture was filtered and washed. According to the above reaction conditions, the activated Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH and octadecanediol mono-tert-butyl ester were coupled on the resin, Fmoc was removed, and the mixture was washed with DCM and dried to obtain P015 peptide resin:

[0177] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-I le-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(tBuO-Ste-γ-Glu(α-OtBu)-γ-Glu(α-OtBu)-γ-Glu(α-OtBu )-AEEA-AEEA)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink-Linker-Nle-MBHA Resin.

[0178] Take P015 peptide resin, add 12-15mL / g peptide resin cleavage agent (TFA:EDT:TIS:H2O, volume ratio 94:2:2:2), stir and react at 25±5℃ for 4 hours, filter the reaction mixture using a sand core funnel, collect the filtrate, wash the resin with a small amount of TFA three times, combine the filtrates and concentrate under reduced pressure, add methyl tert-butyl ether (MBTE) to precipitate, and then wash with MBTE 3-4 times. After evaporating MTBE, the crude product is dried under reduced pressure at room temperature to constant weight to obtain crude P015. After subsequent purification and drying, a P015 sample (mass spectrum MS: 5255.2) is obtained.

[0179] Example 2:

[0180] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0181] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 19).

[0182] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structure has been unfolded.

[0183] The preparation method adopts the Fmoc solid phase peptide synthesis strategy, and the method includes:

[0184] (1) Synthesis of peptide resin intermediate 2

[0185] Take Fmoc-Rink Linker-Nle-MBHA resin (S = 0.49 mmol / g), swell it with an appropriate amount of DCM, and wash it with DCM 2-3 times. Deprotect it with 20% PIP / DMF solution for 30 minutes, wash and filter to obtain the Fmoc-free NH2-Rink linker-Nle-MBHA resin, and drain the solvent for use.

[0186] Take 4 equivalents of Fmoc-Lys(Boc)-OH and HOBt respectively, dissolve them in appropriate amount of DMF / DCM; take another 4 equivalents of DIC, dilute it half with DCM, slowly add it to the DMF / DMC solution with stirring, and react with stirring at -5~0℃ for not less than 60 minutes. After activation, set aside.

[0187] Add the activated Fmoc-Lys(Boc)-OH solution to the NH2-Rink linker–Nle-MBHA resin, control the reaction temperature at 10-30°C, and conduct the coupling reaction for 240–480 min. Filter and wash to obtain the Fmoc-Lys(Boc)-Rink linker–Nle-MBHA resin. Deprotect the resin with a 20% PIP / DMF solution for 30 min, filter and wash to obtain the de-Fmoc Lys(Boc)-Rink linker–Nle-MBHA resin.

[0188] Under the same reaction conditions, start from the second amino acid at the C-terminus and couple to the N-terminus one by one. If the coupling is incomplete (color reaction), use HBTU / DIEA for a second condensation to ensure that each amino acid is completely condensed. The coupling sequence is Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH.H2O, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly- Gly-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH.H2O, Fmoc-Ala-OH.H2O , Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH.H2O, Fmoc-Gln(Trt)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Aib-OH, Fmoc-Ile-OH, Fmoc-Aib-OH, Fmoc-Tyr( tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-α-Me-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Glu(OtBu)-Gly-OH, Fmoc-Aib-OH, Boc-Tyr(tBu)-OH.

[0189] get:

[0190] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-A ib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Le u-Lys(Mtt)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin

[0191] After the above condensation is complete, the Mtt protection is removed with 50% HFIP / DCM solution for 30 minutes, followed by washing and filtration to obtain the peptide resin intermediate 2:

[0192] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala- Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu-Lys-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin.

[0193] (2) Modification chain modification step

[0194] Dissolve 4 equivalents of Fmoc-AEEA-OH and HOBt in an appropriate amount of DMF / DCM. Dissolve another 4 equivalents of DIC in DCM by half, then slowly add it to the DMF / DCM solution with stirring. Stir and react at -5 to 0°C for at least 60 minutes. Activate and set aside.

[0195] The activated Fmoc-AEEA-OH solution was added to the previously swollen and washed peptide resin intermediate 2. The reaction temperature was controlled at 10-30°C. The coupling reaction was carried out for 240-480 minutes. The mixture was filtered and washed. The Fmoc protection was removed with a 20% PIP / DMF solution for 30 minutes. The mixture was filtered and washed. According to the above reaction conditions, the activated Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH and octadecane dioic acid mono-tert-butyl ester were coupled on the resin in sequence. The Fmoc protection was removed and the mixture was washed with DCM and dried to obtain the P019 peptide resin:

[0196] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(t Bu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-V al-Lys(tBuO-Ste-γ-Glu(α-OtBu)-AEEA-AEEA-AEEA)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink-Linker-Nle-MBHA Resin.

[0197] Take P019 peptide resin, add 12-15mL / g peptide resin cleavage agent (TFA:EDT:TIS:H2O, volume ratio 94:2:2:2), stir and react at 25±5℃ for 4 hours, filter the reaction mixture using a sand core funnel, collect the filtrate, wash the resin with a small amount of TFA three times, combine the filtrates and concentrate under reduced pressure, add methyl tert-butyl ether (MBTE) to precipitate, and then wash with MBTE 3-4 times. After evaporating MTBE, the crude product is dried under reduced pressure at room temperature to constant weight to obtain crude P019. After subsequent purification and drying, a P019 sample (mass spectrum MS: 5142.0) is obtained.

[0198] Example 3

[0199] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0200] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 7).

[0201] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K24, for which the structure has been unfolded.

[0202] The peptide of SEQ ID NO: 7 of the present invention (mass spectrum MS: 4997.2) was synthesized using peptide resin intermediate 1 in a similar manner as described in Example 1 above. The difference was that the modification process for the modified chain involved sequentially coupling Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, and octadecandioic acid mono-tert-butyl ester on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain the P007 peptide resin. The other steps were similar.

[0203] Example 4:

[0204] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0205] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 18 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 13).

[0206] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K24, for which the structure has been unfolded.

[0207] Similar to Example 1 above, peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 13 of the present invention (mass spectrum MS: 5025.2). The difference was that the modification process of the modified chain was to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, and mono-tert-butyl eicosadioate on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain the P013 peptide resin. The other steps were similar.

[0208] Example 5:

[0209] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0210] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 14).

[0211] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K24, for which the structure has been unfolded.

[0212] Similar to Example 1 above, peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 14 of the present invention (mass spectrum MS: 5052.4). The difference was that the modification process of the modified chain was to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, and mono-tert-butyl docosanediolate on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain the P014 peptide resin. The other steps were similar.

[0213] Example 6:

[0214] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0215] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 16).

[0216] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K24, for which the structure has been unfolded.

[0217] Similar to Example 1 above, peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 16 of the present invention (mass spectrum MS: 5198.0). The difference was that the modification process of the modified chain was to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, and mono-tert-butyl docosanediolate on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain the P016 peptide resin. The other steps were similar.

[0218] Example 7:

[0219] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0220] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 8).

[0221] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structure has been unfolded.

[0222] Similar to Example 2 above, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 8 of the present invention (mass spectrum MS: 4996.8). The difference was that the modification process of the modified chain was to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, and octadecandioic acid mono-tert-butyl ester on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain P008 peptide resin. The other steps were similar.

[0223] Example 8:

[0224] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0225] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 17).

[0226] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structure has been unfolded.

[0227] Similar to Example 2 above, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 17 of the present invention (mass spectrum MS: 5052.8). The difference was that the modification process of the modified chain was to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, and mono-tert-butyl docosanediolate on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain the P017 peptide resin. The other steps were similar.

[0228] Example 9:

[0229] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0230] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)3-CO-(CH2) 16 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 18).

[0231] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structure has been unfolded.

[0232] The peptide of SEQ ID NO: 18 of the present invention (mass spectrum MS: 5255.2) was synthesized using peptide resin intermediate 2 in a similar manner as described in Example 2 above. The difference was that the modification process for the modified chain involved sequentially coupling Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH, and octadecandioic acid mono-tert-butyl ester on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain the P018 peptide resin. The other steps were similar.

[0233] Example 10:

[0234] Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK;

[0235] wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 20).

[0236] The above structure contains the standard single-letter amino acid code except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structure has been unfolded.

[0237] Similar to Example 2 above, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 20 of the present invention (mass spectrum MS: 5198.4). The difference was that the modification process of the modified chain was to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH and mono-tert-butyl docosane dioate on the resin, followed by Fmoc deprotection, washing with DCM, and drying to obtain P020 peptide resin. The other steps were similar.

[0238] In some embodiments of the present invention, peptide resin intermediate 3 (a peptide resin intermediate with a modified chain at position 16 Lys) is obtained by following the preparation method of peptide resin intermediate 1 in Reference Example 1:

[0239] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys-Gln(Trt)-Ala-Gln(T rt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin

[0240] Peptide resin intermediate 4 (peptide resin intermediate with a modified chain at position 40 Lys) was obtained by following the preparation method of peptide resin intermediate 1 in Reference Example 1:

[0241] Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala- Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys-Rink Linker-Nle-MBHA Resin

[0242] Other dual agonist compounds in the present invention can be prepared by referring to the above method.

[0243] Related assays

[0244] The conditions and data used in the examples for several assays are provided below.

[0245] 1. In vitro function

[0246] (I) In vitro binding activity to human GLP-1 and GIP receptors

[0247] The in vitro binding potency of the compounds of the present invention to human GIP and GLP-1 receptors was evaluated by measuring the binding affinity Ki using crude cell membranes obtained from clonal cell lines overexpressing human GLP-1R cDNA or human GIP-R cDNA.

[0248] 1) In vitro binding activity to human GLP-1 receptor

[0249] hGLP-1 and the compounds of the present invention were dissolved in DMSO and stored at -80°C. 89 μL of membrane (5 μg / well) dissolved in binding buffer (50 mM Hepes, pH 7.4, 5 mM MgCl2, 5 mM EDTA, 0.005% TWEEN, 0.005% HSA) was transferred to a 96-well assay plate. The compounds were serially diluted in DMSO, and then 1 μL of the diluted compound or 100% DMSO was added to the assay plate containing the membrane solution. 10 μL of [ 125 I] GLP-1 (final reaction concentration 0.15 nM). The assay plates were incubated at room temperature for 90 minutes. Membrane complexes were harvested using a Cell Harvester onto GF / B plates pre-coated with 0.5% PEI and rinsed three times with 500 μL of 4°C pre-cooled elution buffer (50 mM Hepes, pH 7.4, 500 mM NaCl). After drying at 37°C for 2 hours, 50 μL of scintillation fluid was added to each well, sealed, and incubated for at least 1 hour. The membrane-bound radioligand levels were then determined using a Microbet2 reader.

[0250] pass[ 125 I] Nonlinear regression of the percentage of GLP-1 binding and the concentration of added compound was used to obtain the absolute IC 50 Concentration. Use the Cheng-Prusoff formula to convert IC 50 The concentration was converted to Ki (Ki is the inhibition constant).

[0251] 2) In vitro binding activity to human GIP receptor

[0252] hGIP and the compounds of the present invention were dissolved in DMSO and stored at -80°C. 98 μL of membrane (15 μg / well) dissolved in binding buffer (50 mM HEPES pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.1% BSA, 0.005% Tween-20) was transferred to a 96-well assay plate. The compounds were serially diluted in DMSO, and then 2 μL of the diluted compound or 100% DMSO was added to the assay plate containing the membrane solution. 100 μL of [ 125 I] GIP (final reaction concentration 0.0315 nM). The assay plate was incubated at room temperature for 90 minutes. Membrane complexes were harvested using a Cell Harvester onto GF / B plates pre-coated with 0.5% PEI and rinsed three times with 500 μL of 4°C pre-cooled elution buffer (50 mM Tris-HCl pH 7.4, 125 mM NaCl). After drying at 37°C for 2 hours, 50 μL of scintillation fluid was added to each well, sealed, and incubated for at least 1 hour. The membrane-bound radioligand levels were then determined using a Microbeta2 reader.

[0253] pass[ 125 The absolute IC was obtained by nonlinear regression of the percentage of GIP binding and the concentration of the added compound. 50 Concentration. Use the Cheng-Prusoff formula to convert IC 50 The concentration was converted to Ki (Ki is the inhibition constant).

[0254] Table 1. Receptor binding affinities, Ki ratios

[0255] NA: Not detected

[0256] Ki ratio: the ratio of the Ki value of the endogenous ligand to the Ki value of the test compound

[0257] The binding affinity of all compounds to receptors is expressed as Ki ratio, with a larger Ki ratio indicating a stronger binding affinity of the compound. Compared with P001, the binding affinity of the compounds of the present invention to human GLP-1R and GIPR is decreased to a certain extent (see Table 1).

[0258] Compared with Tirzepatide, except for P015, the binding affinity of the other nine compounds of the present invention to human GLP-1R is slightly stronger than that of Tirzepatide, and among them, the binding affinity of five compounds, P008, P013, P014, P016 and P019, to GIPR is also slightly stronger than that of Tirzepatide (see Table 1).

[0259] In summary, the compounds of the present invention with different forms of modified chains have a binding affinity to human GLP-1R and GIPR that is somewhat lower than that of P001, but is slightly stronger than or equivalent to that of Tirzepatide.

[0260] (II) Agonistic activity on hGLP-1R and hGIPR

[0261] For human GLP-1 and GIP receptors, the in vitro functional activity of the compounds of the invention at these receptors was determined in HEK-293 clonal cell lines expressing these receptors.

[0262] 1) Agonist activity on human GLP-1 receptor (cAMP reporter gene method)

[0263] The cAMP reporter gene assay was used to determine the agonist activity of the compounds of the present invention on human GLP-1R, with P001, Tirzepatide and endogenous ligand GLP-1 as references.

[0264] HEK293 / CRE / GLP-1R cells were seeded at 50,000 cells / well (80 μL / well) in a 96-well plate and cultured overnight in a 37°C, 5% CO2 incubator. 20 μL / well of assay medium (DMEM containing 0.1% casein) containing a compound (a compound of the present invention, P001, Tirzepatide, or GLP-1) was added to the 96-well plate and incubated in a 37°C, CO2 incubator for another 6 hours. After equilibration to room temperature, the supernatant was removed, and 50 μL / well of Bright-Glo reagent was added. The cells were shaken and lysed at room temperature for 10 minutes. Luminescence was read using an Envision microplate reader to measure luciferase activity.

[0265] The response value of 100 nM GLP-1 was set as 100% response value, and nonlinear regression was performed using GraphPad based on the response percentage and the added compound concentration to obtain the EC of each compound. 50 value.

[0266] 2) Agonist activity on human GIP receptor (LANCE Ultra cAMP assay)

[0267] The LANCE Ultra cAMP Kit was used to determine the agonist activity of the compounds of the present invention on human GIPR, with P001, Tirzepatide and endogenous ligand GIP as references.

[0268] The in vitro agonist activity of the compounds of the present invention against GIPR was determined in HEK293 cells stably expressing human GIPR (HEK293 / GIPR cells). HEK293 / GIPR cells were prepared with HBSS buffer (0.1% Casein, 500 μM IBMX, 5 mM HEPES) and seeded at 1000 cells / well (5 μL / well) in 384-well cell culture plates. 5 μL of HBSS buffer containing 2× compound was added to the 384-well cell culture plates. The plates were sealed and incubated in a 37°C, 5% CO2 incubator for approximately 30 minutes. After incubation, 5 μL of cAMP-Eu working solution and 5 μL of cAMP-Ulight working solution were added, followed by shaking to mix. The plates were incubated at 25°C for 1 hour. The signals at 665 nm and 615 nm were read on an Envision microplate reader. The 665 nm / 615 nm ratio was calculated and converted to cAMP concentration using a cAMP standard curve. The response value of 1 μM GIP was set as 100% response value, and nonlinear regression was performed using GraphPad based on the response percentage and the concentration of the added compound to obtain the EC value of each compound. 50 value.

[0269] Table 2. Agonist activity at human GLP-1 and GIP receptors, EC 50 value

[0270] NA: Not detected

[0271] EC 50 Value: Endogenous ligand EC 50 EC values ​​of the tested compounds 50 Value ratio

[0272] The agonist activities of all compounds on human GLP-1 and GIP receptors were expressed as EC 50 The value indicates that EC 50 The larger the value, the stronger the activity of the compound. Compared with P001, the agonist activity of the compound of the present invention on human GLP-1R and GIPR is decreased to a certain extent (see Table 2).

[0273] Compared with Tirzepatide, all compounds of the present invention have stronger agonist activity on human GLP-1R than Tirzepatide, and among them, six compounds P007, P008, P013, P016, P018 and P019 also have slightly stronger agonist activity on human GIPR than Tirzepatide (see Table 2).

[0274] In summary, the compounds of the present invention with different forms of modified chains have different degrees of reduced agonist activity on human GLP-1R and GIPR compared with P001, but 6 compounds are more potent than Tirzepatide.

[0275] In vitro functional assays showed that the activities of the 10 compounds of the present invention were similar, so a single-dose hypoglycemic experiment in db / db mice was used to further screen the compounds.

[0276] 2. Pharmacokinetics

[0277] (I) Single-dose pharmacokinetics in SD rats

[0278] Plasma was collected from SD rats after subcutaneous or intravenous administration and its plasma drug concentration was measured to illustrate the pharmacokinetic properties of the compounds of the present invention in vivo. The compound was dissolved in phosphate buffer and filtered through a filter membrane (PTFE, 0.45 μm) to obtain a 25 nmol / mL compound solution. Male SD rats (230-268 g, n=3) were given a dose of 50.0 nmol / kg subcutaneously (sc) or 25.0 nmol / kg intravenously (iv). Approximately 150 μL of whole blood was collected from the internal jugular vein in an EDTA-K2 anticoagulant tube at predose, 0.0833 (iv), 0.25, 0.5, 1, 2, 4, 6, 8, 24, 48, 72, 96, and 120 h. The blood samples were centrifuged at 1500 g for 10 min to obtain plasma, which was stored frozen at -90°C to -60°C for analysis.

[0279] Sample processing

[0280] (1) Sample processing methods for Tirzepatide, P007, P008, P013, and P014

[0281] Take 30.0 μL of thawed plasma sample and add 150 μL of acetonitrile solution (containing 5 ng·mL -1 Verapamil, 50 ng mL -1 Glyburide, 200 ng·mL -1 Tolbutamide and 200 ng·mL -1 Protein was precipitated with diclofenac (diclofenac), the mixture was vortexed for 5 minutes, and centrifuged at 3700 rpm for 8 minutes. 70.0 μL of the supernatant was added to 70.0 μL of 0.5% formic acid solution, and the mixture was vortexed for 5 minutes. 15 μL of the mixture was sampled and injected into LC-MS / MS to measure the plasma drug concentration.

[0282] (2) Sample processing method for P015, P016, P017, and P018:

[0283] Take 20.0 μL of thawed plasma sample and add 60.0 μL of acetonitrile solution (containing 5 ng·mL -1 Verapamil, 50 ng mL -1 Glyburide, 200 ng·mL -1 Tolbutamide and 200 ng·mL -1 Protein was precipitated with diclofenac (diclofenac), the mixture was vortexed for 1 minute, and centrifuged at 13,000 rpm for 8 minutes. 60.0 μL of the supernatant was added to 60.0 μL of 0.5% formic acid solution, and the mixture was vortexed for 10 minutes. 10.0 μL of the mixture was sampled and injected into LC-MS / MS for plasma drug concentration determination.

[0284] (3) Sample processing method for P019 and P020:

[0285] Take 30.0 μL of thawed plasma sample and add 150 μL of acetonitrile solution (containing 5 ng·mL -1 Verapamil, 50 ng mL -1 Glyburide, 200 ng·mL -1 Tolbutamide and 200 ng·mL -1 Protein was precipitated with diclofenac (diclofenac), the mixture was vortexed for 5 minutes, and centrifuged at 3700 rpm for 8 minutes. 70.0 μL of the supernatant was added to 70.0 μL of 0.5% formic acid solution, and the mixture was vortexed for 5 minutes. 10.0 μL of the mixture was sampled and injected into LC-MS / MS to measure the plasma drug concentration.

[0286] Based on the blood concentrations of the compounds in SD rats, the pharmacokinetic parameters were calculated using the Winnolin 8.2 software with a non-compartmental model. The results are shown in Tables 3 and 4 below.

[0287] Table 3 Pharmacokinetic parameters of different peptide compounds after subcutaneous injection in rats

[0288] Table 4 Pharmacokinetic parameters of different polypeptide compounds after subcutaneous injection in rats

[0289] PK studies of subcutaneous injections of the compounds of this invention in rats demonstrated that the half-lives of P007, P008, P013, P014, P015, P016, P017, P018, and P020 were greater than that of Tirzepatide (increases of approximately 23.7% to 64.0%). The modified chains of the compounds of this invention achieved longer plasma half-lives than Tirzepatide, the marketed dual-target product. Furthermore, compared to P001, the half-lives of the compounds of this invention were significantly prolonged, increasing by more than 15-fold, 20-fold, and even 30-fold.

[0290] (II) Pharmacokinetics in C57 mice

[0291] After subcutaneous or intravenous administration to C57 mice, plasma was collected and its blood drug concentration was measured to illustrate the pharmacokinetic properties of the compounds of the present invention in vivo. The compound was dissolved in phosphate buffer containing 0.1% Tween 20 and filtered through a filter membrane (PTFE, 0.45μm) to obtain a 15nmol / mL compound solution. C57 mice (18-22g) were given a dose of 30nmol / kg subcutaneously (sc) or 15.0nmol / kg intravenously (iv). At predose, 0.5, 1, 2, 4, 8, 24, 48 and 72h, approximately 100μL of whole blood was collected from the eye socket in an EDTA-K2 anticoagulant tube. The blood sample was centrifuged at 1500g for 10min to obtain plasma, which was frozen at -90°C to -60°C for analysis and testing.

[0292] Sample processing

[0293] (1) Sample processing methods for Tirzepatide, P014, P016, and P020

[0294] Take 20.0 μL of thawed plasma samples and add 20 μL of 50% methanol aqueous solution (containing 100 ng·mL -1 The protein was precipitated with 60 μL of acetonitrile (internal standard) and vortexed for 5 minutes, followed by centrifugation at 3700 rpm for 8 minutes. 50.0 μL of the supernatant was added to 50.0 μL of 0.5% formic acid solution, vortexed for 5 minutes, and 50 μL of the mixture was injected into the sample for LC-MS / MS analysis of plasma drug concentrations.

[0295] Based on the blood concentrations of the compounds in C57 mice, the pharmacokinetic parameters were calculated using the non-compartmental model using Winnolin 8.2 software. The results are shown in Tables 5 and 6 below.

[0296] Table 5 Average pharmacokinetic parameters after single subcutaneous administration in mice

[0297] Table 6 Average pharmacokinetic parameters after single intravenous administration in mice

[0298] PK studies of the compounds of this invention in mice following subcutaneous injection showed that the half-lives of P014, P016, and P020 were significantly longer than that of Tirzepatide (increases of approximately 59.2% to 92.5%). The compounds of this invention modified with modified chains can achieve longer plasma half-lives than Tirzepatide, a marketed dual-target product.

[0299] (III) Pharmacokinetics in cynomolgus monkeys

[0300] Plasma concentrations were collected from cynomolgus monkeys after subcutaneous or intravenous administration to elucidate the pharmacokinetic properties of the compounds of this invention. The compounds were dissolved in 1.4% PG in 8mM Na2HPO4 buffer (pH 7.39) and filtered through a 0.45μm PTFE membrane to obtain 25 and 12.5 nmol / mL solutions, respectively. Cynomolgus monkeys (4-8 kg) were administered 25 nmol / kg subcutaneously (sc) or 12.5 nmol / kg intravenously (iv). Blood was collected via the cephalic vein or other appropriate vein at predose, 2 h, 12 h, 24 h, 32 h, 48 h, 3 d (72 h), 4 d (96 h), 6 d (144 h), 9 d (216 h), 12 d (288 h), 15 d (360 h), and 18 d (432 h). Approximately 500 μL of whole blood was collected in EDTA-K2 anticoagulant tubes. Blood samples were centrifuged at 2200 g for 10 min to obtain plasma, which was frozen at -90°C to -60°C for analysis.

[0301] Sample processing

[0302] (1) Sample processing methods for Tirzepatide, P014, P016, P017, and P020

[0303] 70 μL of thawed plasma sample was collected and protein was precipitated with 70 μL of acetonitrile (containing 1 ng / mL internal standard). The mixture was vortexed for 1 minute and centrifuged at 14,000 rpm for 10 minutes. 60 μL of the supernatant was added to 60 μL of 0.5% formic acid solution and vortexed for 5 minutes. 10 μL of the mixture was sampled and injected into LC-MS / MS for plasma drug concentration analysis.

[0304] Based on the plasma concentrations of the compound in cynomolgus monkeys, the pharmacokinetic parameters were calculated using the Winnolin 8.2 software with a non-compartmental model. The results are shown in Tables 7 and 8 below.

[0305] Table 7 Mean pharmacokinetic parameters after single subcutaneous administration in cynomolgus monkeys

[0306] Table 8 Mean pharmacokinetic parameters after single intravenous administration in cynomolgus monkeys

[0307] PK studies of subcutaneous injections of the compounds of this invention in cynomolgus monkeys demonstrated that the half-lives of P014, P016, and P020 were comparable to those of Tirzepatide, while the Tmax values ​​of P016 and P017 were extended compared to Tirzepatide. The compounds of this invention modified with modified chains can achieve plasma half-lives comparable to those of Tirzepatide, a marketed dual-target product. The Tmax values ​​of P016 and P017 are approximately twice that of Tirzepatide, and the time to peak drug concentration is significantly delayed compared to Tirzepatide, potentially leading to milder / less gastrointestinal side effects or shorter titration times.

[0308] Pharmacodynamics

[0309] (I) Pharmacodynamics in db / db mice: To study the effects of the compounds of the present invention on blood glucose in diabetic model mice (db / db mice)

[0310] This study used a single subcutaneous administration of the compound to db / db mice. Changes in blood glucose, food intake, and body weight were measured to elucidate the hypoglycemic effects and duration of efficacy of the compound described herein, and compared with positive controls Tirzepatide and P001. Male db / db mice, 8-9 weeks of age, were used in this study. The db / db mice were housed in individually ventilated cages in an IVC facility with controlled temperature (20-26°C) and humidity (40-70%), a 12h:12h light / dark cycle, and free access to food and water. Blood was collected from the tail tip of the mouse using a Roche blood glucose meter to measure basal blood glucose. The mice were randomly divided into groups (n=6 / group) based on their initial blood glucose and initial body weight. All groups had similar body weights and blood glucose levels.

[0311] The compound of the present invention (10 nmol / kg) or the positive control Tirzepatide (10 nmol / kg) or P001 (10 nmol / kg) were dissolved in a solvent (PBS containing 0.1% Tween 20, pH 7.2-7.4). After a single subcutaneous administration, random blood glucose levels were recorded at set time points (0-120 hours, recording of random blood glucose for the compound was stopped when the random blood glucose level was no different from that of the vehicle group), as well as daily body weight and food intake. Data were statistically analyzed using GraphPad Prism 8. Statistical differences between groups were analyzed using T-TEST, and differences were considered significant when P < 0.05.

[0312] Duration of hypoglycemic effect: the longest time during which the random blood glucose level of the compound is significantly different from that of the Vehicle group (P<0.05).

[0313] Table 9. Duration of hypoglycemic effect of compounds

[0314] Compared with P001, the glucose-lowering effect of the compounds of the present invention lasts 24 hours or longer (P007, P008, P014, P019), even longer than 40 hours (P008, P014), and longer than 72 hours (P014) (see Table 9, Figure 1A, Figure 1B).

[0315] Compared with Tirzepatide, the duration of blood sugar lowering effect of the compounds of the present invention is comparable to or longer (P007, P008, P013, P014, P016, P017, P018, P019, P020), even 16 hours or more (P008, P013, P014, P016, P017, P020), 24 hours or more (P013, P014, P016, P017, P020), 48 hours or more (P014, P016, P020), and 56 hours or more (P016, P020) (see Table 9).

[0316] Compared with P001: Based on the test described above, during the experimental period, P008 (p < 0.01), P014 (p < 0.001), and P019 (p < 0.05) showed significantly better glucose-lowering efficacy than P001 (blood glucose AUC, P < 0.05, as shown in Figure 2A). The glucose AUC inhibition rates were 2.3, 2.6, and 2.2 times that of P001, respectively. The glucose-lowering durations were 40, 72, and 24 hours longer than those of P001, respectively (Table 9). P007 had a glucose-lowering effect comparable to that of P001, but the duration was 24 hours longer than that of P001 (as shown in Figure 2A and Table 9).

[0317] Compared with Tirzepatide: P014 (p < 0.05, Figure 2B), P016 (p < 0.05, Figure 2C), and P020 (p < 0.05, Figure 2C) showed stronger glucose-lowering effects (blood glucose AUC, p < 0.05) than Tirzepatide, with glucose AUC inhibition rates of 1.7, 2.2, and 2.0 times that of Tirzepatide, respectively. Furthermore, they exhibited significantly longer durations of action (48, 56, and 56 hours longer than Tirzepatide, respectively; Table 9). P013 and P017 had comparable glucose-lowering effects to Tirzepatide (Figure 2C), but their duration of action was 24 hours longer than that of Tirzepatide (Table 9).

[0318] Surprisingly, compared to P001, the acylated GLP-1 / GIP agonists of this invention significantly enhance the glucose-lowering effect while achieving a long-lasting effect. Compounds of this invention with various chain modifications can achieve stronger and longer-lasting glucose-lowering effects than P001, and some compounds of this invention exhibit stronger and longer-lasting glucose-lowering effects than Tirzepatide.

[0319] Furthermore, in combination with the target binding and agonist activity of the compounds of the present invention on human GLP-1 and GIP receptors, it was unexpectedly found that the target binding and agonist activity of the compounds of the present invention on the human GLP-1 receptor were significantly reduced compared with those of P001, and the target binding and agonist activity of the compounds of the present invention on the human GIP receptor were also significantly reduced, but the hypoglycemic effect of the compounds of the present invention was significantly better than that of P001.

[0320] (II) Multiple-dose experiment in db / db mice

[0321] In this study, the compounds of the present invention were administered subcutaneously multiple times to db / db mice, and changes in the blood glucose of the mice were detected to further illustrate the hypoglycemic effect of the compounds of the present invention and to compare with the positive control Tirzepatide. In this study, male db / db mice aged 7-8 weeks were used. The db / db mice were housed in an IVC facility with controlled temperature (20-26°C) and humidity (40-70%), a 12h:12h light / dark cycle, and free access to food and water. Blood was collected from the tip of the tail, and basal blood glucose was measured with a Roche blood glucose meter. The mice were randomly divided into groups (n=6 / group) based on initial blood glucose and initial body weight, and each group had similar body weight and blood glucose.

[0322] The compound of the present invention (3, 10, 30 nmol / kg) or the positive control Tirzepatide (3, 10, 30 nmol / kg) was dissolved in a solvent (PBS containing 0.1% Tween20, pH 7.2-7.4). The drug was administered by subcutaneous injection once every 3 days for four weeks. Subcutaneous injection was performed on days 1, 4, 7, 10, 13, 16, 19, 22 and 25. Random blood glucose was recorded every 3 days throughout the study. Fasting blood glucose was tested on day 28, and then blood was collected by orbital bleeding. After orbital bleeding, the animals were sacrificed and the liver was removed and weighed. The collected blood was centrifuged at 3000 rpm for 10 minutes after 1 to 2 hours to separate the serum. Biochemical indicators such as triglycerides (TG), alanine aminotransferase (ALT), and total bilirubin (TBIL) were detected using a biochemical analyzer. The data results were statistically analyzed using GraphPad Prism8, and the statistical differences between the groups were analyzed using T-TEST. P < 0.05 was considered to be significant.

[0323] In experiments conducted as described above, compounds of the present invention, such as P014, P016, P017, and P020, all demonstrated superior blood glucose suppression compared to Tirzepatide at a 3 nmol / kg dose; at a 30 nmol / kg dose, P016 was more effective than Tirzepatide (see Figures 3A-3D and Table 10). P016, P017, and P020 significantly reduced liver weight in db / db mice at multiple doses, whereas Tirzepatide did not reduce liver weight at any of the three experimental doses. Compounds of the present invention, such as P014, P016, P017, P020, and Tirzepatide, all effectively reduced serum triglycerides in db / db mice (except in the 3 nmol / kg P017 and Tirzepatide groups), with P016 and P020 exhibiting superior efficacy to Tirzepatide in some dose groups (see Table 11). Furthermore, P014 and P016 also reduced serum TBIL and / or ALT.

[0324] Table 10. Fasting blood glucose and blood glucose AUC of db / db mice

[0325] T-test, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 compared with the vehicle group; #p<0.05, ##p<0.01, ###p<0.001 compared with the Tirzepatide (TZP) group at the same dose. Results are expressed as mean ± SEM of 6 mice.

[0326] Table 11. Liver weight and serum TG of db / db mice

[0327] T-test, *p<0.05, **p<0.01, ***p<0.001 compared with the vehicle group; #p<0.05, ##p<0.01 compared with the Tirzepatide (TPZ) group at the same dose. The results are expressed as Mean ± SEM of 6 mice.

[0328] Table 12. Serum ALT and TBIL of db / db mice

[0329] T-Test, *p<0.05 compared with the vehicle group; #p<0.05, ##p<0.01 compared with the Tirzepatide (TPZ) same-dose group.

[0330] The results are expressed as Mean ± SEM of 6 mice.

[0331] In summary, in this experimental system, the compounds of this invention effectively lowered blood glucose in db / db mice after subcutaneous administration for four weeks. The effective dose (≤3 nmol / kg) was one-third that of Tirzepatide (10 nmol / kg). Furthermore, some compounds of this invention, such as P016, exhibited a greater maximum efficacy (30 nmol / kg) than Tirzepatide. Furthermore, some compounds of this invention demonstrated liver weight reduction and liver protection in db / db mice, whereas Tirzepatide did not exhibit similar effects.

[0332] (III) Repeated administration of drugs to male rats for 2 weeks

[0333] 1. Experimental Design

[0334] Seventy-seven male Sprague-Dawley (SPF) rats were randomly divided into 10 groups according to body weight. Group 1 was a vehicle control group consisting of 5 animals. Groups 2-4 were commercially available drug control groups (Tirzepatide), each consisting of 8 animals (5 animals in the main test and 3 animals in the toxicity group), with doses of 1, 3, and 10 mg / kg, respectively. Groups 5-7 were test group 1, and groups 8-10 were test group 2. Each group consisted of 8 animals (5 animals in the main test and 3 animals in the toxicity group), with doses of 1, 3, and 10 mg / kg, respectively. All groups were dosed twice weekly for 2 consecutive weeks (dosing days: Days 1, 5, 8, 12, and 15).

[0335] During the experiment, all groups of animals were subjected to cage observation, detailed clinical observation, food consumption measurement, body weight measurement, blood biochemistry and hematological index measurement, gross anatomical observation, organ weighing, and toxicokinetic study.

[0336] 2. Data Statistics

[0337] The following table details the combinations used for statistical comparison.

[0338] Statistical Methods Table

[0339] Raw data for each time period will be tabulated, and the mean and standard deviation / or category change will be calculated for each endpoint by group and sex. For each endpoint, the treated group will be compared to the control group as described below. Endpoint data will be log-transformed before the start of specific analyses, as required.

[0340] Statistical analysis table

[0341] 3. Experimental Results

[0342] Body Weight / Weight Gain: After the first dose, weight loss was observed in all treatment groups at the following doses: Tirzepatide ≥ 3 mg / kg; P014 and P016 ≥ 1 mg / kg. With continued dosing, body weight recovered, and by the end of dosing, all groups maintained weight gain, but weight gain was lower than that of the control group, demonstrating a clear dose-related relationship.

[0343] Food intake: A dose-related decrease in food intake was observed in all the treatment groups (Tirzepatide, P014 and P016) throughout the entire experiment. Blood biochemistry: Amylase and triglyceride levels decreased. Hematology: Reticulocyte levels were slightly decreased. Gross autopsy: No abnormalities were found in any of the treatment groups (Tirzepatide, P014 and P016). Organ weight: The weight of the heart, liver, spleen, etc. in each treatment group (Tirzepatide, P014 and P016) was reduced, with little difference compared with the vehicle control group, which may be related to weight loss and is considered to be related to pharmacological effects. Toxicity: C in the Tirzepatide, P014 and P016 groups was significantly decreased. max Both the AUC and the TZP level increased in a dose-proportional manner. No significant accumulation was observed in the TZP, P014, and P016 dose groups after administration twice a week for 2 consecutive weeks.

[0344] Each treatment group (Tirzepatide, P014 and P016) was given twice a week, and the MTD of the two-week treatment was ≥10 mg / kg, corresponding to the C max and AUC last And the test safety window results are as follows:

[0345] Table 13. Safety window of each dosing group

[0346] In summary, the toxicity properties of P016 and P014 are similar to those of Tirzepatide, but they have a higher safety window (calculated by exposure, the safety window of P014 is more than 4 times that of Tirzepatide, while the safety window of P016 is more than 7 times that of Tirzepatide).

[0347] In addition, relative to the safety window of P001, the safety window of the compounds of the present invention is 10 times larger, or 20 times larger, or 30 times larger, or 40 times larger, or 50 times larger, or 60 times larger, or 70 times larger, or 80 times larger, or 90 times larger than that of P001, especially P014 and P016.

[0348] At the same time, at the same exposure dose, the effect of the compound of the present invention on HR is lower than that of Tirepatide, the heart rate increase of P016 is lower than that of Tirepatide, and there is no sustained heart rate acceleration of more than 30%. At the same exposure dose, the heart rate recovery of P016 is faster than that of TZP (the recovery time of P016 is between 72 and 120 hours, and the recovery time of TZP is greater than 120 hours).

[0349] The peptide compounds and applications provided by the present invention are introduced in detail above.

[0350] Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The above examples are merely intended to facilitate understanding of the methods and central concepts of the present invention. It should be noted that those skilled in the art may make various improvements and modifications to the present invention without departing from the principles of the present invention, and such improvements and modifications are also protected by the claims of the present invention.

[0351] Amino acid sequence:

Claims

1. A compound of the following formula (AI), or a pharmaceutically acceptable salt thereof: Y-X1-EGT-X2-TSDY-A11-A12-A13-LDK-A17-AQ-A20-EFVKWLLK-A29-GPSSGAPPPSK Formula (AI); wherein X1 is Aib; X2 is αMePhe; A11 is Aib or Ala; A12 is Ala, Ile, Lys, Phe, or Pya (4); A13 is Aib, Cha, Leu, αMePhe, or αMeTyr; A17 is Gln or Ile; A20 is Ala or Ser; A29 is Gln or Gly, 1 K, 2 K, 3 K or 4 K positions are selected from the 16th, 24th, 28th and 40th positions by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by being conjugated to the ε-amino group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 24, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester.

2. The compound according to claim 1, wherein A11 is Aib; or / and A12 is Ile; or / and A13 is Aib; or / and A17 is Gln; or / and A20 is Ala; or / and A29 is Gly; or / and a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, c is independently selected from an integer of 12 to 22; further c is independently selected from 14, 16, 18 or 20.

3. A compound of the following formula (I), or a pharmaceutically acceptable salt thereof: Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; 1, 2, 3 or 4 K positions are selected from positions 16, 24, 28 and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugation to the ε-amino group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 24, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester; and the C-terminal amino acid is amidated to a C-terminal primary amide.

4. The compound according to claim 3, which is: Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I), or a pharmaceutically acceptable salt thereof, wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; one or two K positions are selected from positions 16, 24, 28, and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -Z is chemically modified by conjugation to the ε-amino group of the K side chain, wherein Z is independently selected from -CH3, carboxylic acid or carboxylic acid bioisostere, phosphonate / ester or sulfonate / ester, preferably -CO2H, each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to a C-terminal primary amide.

5. The compound according to claim 4, wherein At position 16, K is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain for chemical modification; or At position 24, K is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain for chemical modification; or At position 28, K is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain for chemical modification; or At position 40, K is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H was conjugated to the ε-amino group of the K side chain for chemical modification.

6. The compound according to claim 4, wherein K at positions 24 and 28 is synthesized by replacing the 2-(2-amino-ethoxy)-ethoxy]-acetyl group with a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains; or K at positions 16 and 24 is synthesized by replacing the acetyl group with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains; or K at positions 16 and 28 was synthesized by replacing the ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains; or K at positions 16 and 40 is synthesized by replacing the acetyl group with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains; or K at positions 24 and 40 is synthesized by replacing the 2-(2-amino-ethoxy)-ethoxy]-acetyl group with a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains; or K at positions 28 and 40 is synthesized by replacing the 2-(2-amino-ethoxy)-ethoxy]-acetyl group with a -(γ-Glu) b -CO-(CH2) c -CO2H was conjugated to the ε-amino group of the K side chain to perform chemical modification of the two modified chains.

7. The compound according to any one of claims 3 to 6, wherein said a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, c is independently selected from an integer of 12 to 22; further c is independently selected from 14, 16, 18 or 20; and / or Z is -CO2H.

8. The compound according to claim 4, wherein the compound is Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I); wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain and chemically modified, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

9. The compound according to claim 4, wherein the compound is Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK formula (I); wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) a -(γ-Glu) b -CO-(CH2) c -CO2H is conjugated to the ε-amino group of the K side chain and chemically modified, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

10. The compound according to any one of claims 8-9, wherein said a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, c is independently selected from an integer of 12 to 22; further c is independently selected from 14, 16, 18 or 20.

11. The compound according to claim 4, wherein the compound is Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 16 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 7); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 18 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 13); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 20 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 14); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)3-CO-(CH2) 16 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 15); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 20 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 16); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 16 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 8); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) 20 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 17); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)3-CO-(CH2) 16 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 18); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 16 -CO2H is chemically modified by conjugation to the ε-amino group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 19); or Y-X1-EGT-X2-TSDY-X3-I-X4-LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; X4 is Aib; K at position 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl)3-(γ-Glu)1-CO-(CH2) 20 The K side chain was chemically modified by conjugating -CO2H to the ε-amino group; and the C-terminal amino acid was amidated to a C-terminal primary amide (SEQ ID NO: 20).

12. The compound according to claim 4, wherein the compound is or a pharmaceutically acceptable salt thereof.

13. A pharmaceutical composition comprising the compound according to any one of claims 1 to 12 and a pharmaceutically acceptable carrier, diluent or excipient.

14. Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or the composition according to claim 13, in the preparation of a medicament for treating or preventing hyperglycemia, impaired glucose tolerance, type I diabetes, type II diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular diseases, stroke, inflammatory bowel syndrome, dyspepsia, and gastric ulcer.

15. Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or the composition according to claim 13, in the preparation of a medicament for delaying or preventing the progression of type II diabetes.

16. Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or the composition according to claim 13, in the preparation of a medicament for reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell mass, and / or restoring β-cell glucose sensitivity.

17. A method for treating type II diabetes, comprising administering an effective amount of the compound according to any one of claims 1 to 12, or the composition according to claim 13, to a patient in need thereof.

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