Dual-agonist compound for both GLP-1 and GIP receptors and application thereof

Abstract

Provided is a dual- agonist compound for both Glucagon-like Peptide-1 (GLP-1 ) and Glucose-dependent Insulinotropic Polypeptide (GIP) receptors and application thereof. In particular, provided are a polypeptide analog derived from GLP-1 and a pharmaceutically acceptable salt thereof, which has an agonist effect on human GLP-1 receptor and human GIP receptor and can be used for the treatment of metabolic diseases such as obesity, type II diabetes, and non- alcoholic fatty liver. FIGURE 1

Description

Dual agonist compounds for GLP-1 and GIP receptors and uses thereof

[0001] The present disclosure claims priority to patent application 202010472577.8 filed on May 29, 2020 and patent application 202110335100.X filed on March 29, 2021. TECHNICAL FIELD

[0002] The present disclosure belongs to the field of biological medicine, in particular relates to a compound having dual agonist effect on human glucagon-like peptide-1 (GLP-1) receptor and human glucose-dependent insulinotropic polypeptide (GIP) receptor and pharmaceutically acceptable salts thereof, and can be used for the treatment of metabolic diseases such as diabetes and / or obesity. BACKGROUND

[0003] Diabetes is a metabolic disease caused by insufficient secretion of insulin in the body, which leads to the disorder of glucose, protein and lipid metabolism in the human body. According to the difference of its pathological mechanism, diabetes is mainly divided into insulin-dependent diabetes (type I diabetes) and non-insulin-dependent diabetes (type II diabetes). Among them, 90-95% of diabetes patients worldwide are non-insulin-dependent diabetes. Non-insulin-dependent diabetes is a long-term chronic metabolic disease caused by impaired function of pancreatic beta cells and long-term insulin resistance, and its most important feature is the lack of insulin level in the body and high concentration of glucose in the blood. Studies have shown that non-insulin-dependent diabetes is associated with a variety of high-risk complications in patients, and often leads to patients suffering from cardiovascular disease, kidney failure, blindness, amputation and other complications.

[0004] One of the main causes of non-insulin-dependent diabetes is obesity. Obesity is defined as excessive or abnormal accumulation of fat in the body that damages human health. According to the body mass index (BMI) of a person, obesity can also be defined as when the BMI index of a person is greater than or equal to 30 kg / m 2 The occurrence of obesity can significantly increase the risk of human suffering from cardiovascular disease, diabetes, musculoskeletal disorders and certain cancers. In addition, the increase of body mass index of a person can also increase the risk of suffering from certain non-communicable diseases.

[0005] Due to the huge number of patients and the significant economic burden caused by diabetes and its complications, the development of safe and effective drugs for the treatment of diabetes has always been the focus of many research institutions and pharmaceutical companies. Currently, the approved diabetes drugs mainly include chemical synthesis of small molecule oral hypoglycemic drugs, such as biguanides, sulfonyl ureas, insulin sensitizers, α-glucosidase inhibitors and injection hypoglycemic drugs produced by biosynthesis, such as recombinant insulin and its derivatives. Although the above drugs can effectively control the blood glucose level in the plasma of diabetic patients in clinical practice, long-term use often accompanies adverse reactions such as weight gain in patients, which in turn leads to an increase in the risk of potential cardiovascular diseases and a decrease in patient compliance. Considering the potential pathological relationship between diabetes and obesity and the potential risk of complications caused by obesity, the development of a drug that can effectively control blood glucose while also appropriately reducing the weight of diabetic patients has multiple implications for the effective treatment of diabetes and the reduction of potential complications, and is therefore a more excellent research direction in clinical practice.

[0006] Glucagon like peptide-1 (GLP-1) is a gastrointestinal regulatory polypeptide containing 30 or 31 amino acid residues. The secretion of GLP-1 is mainly regulated by L-cells on the small intestine according to the absorption of nutrients and fluctuation of blood glucose level in the body. After food intake, L-cells of the small intestine secrete a large amount of GLP-1 to enhance the endocrine function of the pancreas. GLP-1 polypeptide mainly completes its physiological function of controlling blood glucose and reducing appetite in the body by activating GLP-1 receptors distributed on the cell membrane surface. The mechanism of GLP-1 controlling blood glucose level in the body is mainly to activate its GLP-1 receptors distributed in the beta cells of the islets of Langerhans to promote the biosynthesis and secretion of insulin. At the same time, GLP-1 polypeptide can inhibit the secretion of glucagon, gastric emptying and food intake under the condition of high blood glucose level in the body, and enhance the degradation of glucose in the body through specific nervous system effects. It is worth noting that the physiological function of GLP-1 polypeptide promoting insulin secretion is controlled by the concentration of plasma glucose, so compared with other diabetes drugs, GLP-1 polypeptide will not cause severe and persistent hypoglycemia. In addition, it is reported in the literature that GLP-1 polypeptide and its analogs have a direct promoting effect on the growth, differentiation and proliferation of beta cells of experimental animals, indicating that GLP-1 polypeptide and its analogs can have physiological functions of protecting the islets of Langerhans and delaying the progression of diabetes and inhibiting the apoptosis of beta cells. GLP-1 polypeptide also has the potential to inhibit the secretion of gastric acid stimulated by gastrin and food intake. These characteristics mean that GLP-1 polypeptide also has the physiological effect of preventing digestive tract ulcers. GLP-1 polypeptide can also activate GLP-1 receptors distributed in the central nervous system of the brain to enhance satiety, reduce food intake and achieve the physiological effect of maintaining or reducing body weight. Therefore, the wide mechanism of action and physiological functions of GLP-1 polypeptide mean that GLP-1 polypeptide is an ideal drug for treating non-insulin-dependent diabetes and obesity diabetes.

[0007] The physiological functions of GLP-1 polypeptide in controlling blood glucose and reducing body weight bring hope for the treatment of non-insulin-dependent diabetes / obesity diabetes. Natural GLP-1 has poor drug properties, and it is easily degraded by dipeptidyl peptidase-IV (DPP-IV) in the body, so that its half-life in the human body is only 1-2 minutes. In the face of this difficulty, the pharmaceutical industry has constructed long-acting GLP-1 analogs and derivatives by performing site-directed mutagenesis of the amino acids at the enzyme cleavage site, fatty acid modification of the polypeptide backbone, and combination of GLP-1 polypeptide with various proteins / high molecular weight polymers. The long-acting GLP-1 analogs that have been marketed and widely used in clinical practice at this stage include once-daily subcutaneous injection of liraglutide and once-weekly subcutaneous injection of dulaglutide and semaglutide, etc.

[0008] In the clinic, the side effects of GLP-1 polypeptides and their derivatives are mainly manifested in nausea, vomiting and diarrhea caused by the gastrointestinal tract. In addition, it has been found that GLP-1 polypeptides and their derivatives can also cause the heartbeat of the subject to accelerate and, in certain cases, increase the risk of pancreatitis in patients. Therefore, the dosage of GLP-1 polypeptides and their derivatives is limited by the side effects they cause, and thus the clinical use thereof cannot achieve full blood glucose control and weight loss in patients.

[0009] Glucose-dependent insulinotropic polypeptide (GIP) and GLP-1 polypeptide both belong to the incretins, which play a key physiological role in the metabolism of blood glucose in the body. GIP mainly consists of 42 amino acid residues in the body and is secreted by the duodenum and the K cells near the jejunum according to the glucose level in the plasma. GIP polypeptide exerts its physiological effect by binding to the GIP receptor distributed in the pancreatic beta cells, adipose tissue and central nervous system. Similar to GLP-1 polypeptide, GIP polypeptide can stimulate the pancreatic beta cells to secrete insulin, thereby reducing the blood glucose concentration in the plasma and protecting the pancreatic beta cells to control the metabolism of glucose in the body. In addition, the physiological function of GIP polypeptide also includes activating the GIP receptor in adipose tissue to promote the metabolism of fat. Intracerebroventricular injection of GIP polypeptide in mice can reduce food intake and body weight in the test animals, which seems to indicate that GIP polypeptide also has a physiological function in reducing body weight. Studies have shown that in non-insulin-dependent diabetic patients, the incretin function of GIP polypeptide is greatly reduced, resulting in a lack or loss of incretin effect in patients. Studies have shown that the inhibitory effect of GIP polypeptide produced by these diabetic patients is greatly reduced when the blood glucose level returns to normal.

[0010] Therefore, there is a clinical need for a method of using GIP polypeptide to treat non-insulin-dependent diabetes and a clinically effective hypoglycemic drug combination to restore the tolerance of non-insulin-dependent diabetic patients to GIP polypeptide and further combine the incretin effect of GIP polypeptide to achieve a stronger clinical hypoglycemic effect.

[0011] The purpose of the present disclosure is to provide a derivative of GLP-1 analogs having agonist activity on human GIP receptors, which has a dual agonist effect on human GLP-1 receptors and human GIP receptors. In addition, some compounds of the present disclosure have a stronger therapeutic effect on reducing blood glucose and reducing body weight compared to GLP-1 receptor agonists known in the art. Some compounds of the present disclosure have very high plasma stability and have pharmacokinetic characteristics of once-a-week subcutaneous injection in human subjects.

[0012] SUMMARY

[0013] The present disclosure provides a GLP-1 analogue having the general formula (I), or a pharmaceutically acceptable salt thereof:

[0014] R1-X1-X2-Glu-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -Glu-Phe-X 23 -X 24 -Trp-Leu-X 27 -X 28 -X 29 -X 30 -Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2

[0015] (I) (SEQ ID NO: 19)

[0016] wherein:

[0017] R1is hydrogen (H), alkyl, acetyl, formyl, benzoyl, trifluoroacetyl, pGlu or absent;

[0018] R2is -NH2, -OH or absent;

[0019] X1, X2, X 10 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 23 , X 24 , X 27 , X 28 , X 29 and X 30 are independently selected from any natural amino acid residue, non-natural amino acid residue or a peptide segment consisting of natural amino acid residues and / or non-natural amino acid residues.

[0020] Some embodiments of this disclosure provide GLP-1 analogs having the general formula (I) or pharmaceutically acceptable salts thereof, wherein X1 is selected from amino acid residues of Tyr or His; X2 is selected from amino acid residues of Aib or D-Ala; X 10 Amino acid residues selected from Val or Tyr; X 12 Amino acid residues selected from Ser or Ile; X 13 Amino acid residues selected from Tyr or Ala; X 14 Amino acid residues selected from Leu or Nle; X 15 Amino acid residues selected from Asp or Glu; X 16 Amino acid residues selected from Arg, Glu, Gly, Lys, or Aib; X 17 Amino acid residues selected from Glu, Ile, or Gln; X 18 Amino acid residues selected from Ala, Aib, or His; X 19 Amino acid residues selected from Ala, Aib, or Gln; X 20 Amino acid residues selected from Gln, Glu, and Lys; X 23 Amino acid residues selected from Ile or Val; X 24 Amino acid residues selected from Ala, Asn, or Gln; X 27 Amino acid residues selected from Val or Leu; X 28 Amino acid residues selected from Arg or Ala; X 29 Amino acid residues selected from Gly or Gln; X 30 The amino acid residues are selected from Gly and Lys.

[0021] Some embodiments of this disclosure provide GLP-1 analogs having general formula (I) or pharmaceutically acceptable salts thereof, wherein X1 is selected from amino acid residues of Tyr or His; X2 is selected from amino acid residues of Aib or D-Ala; X 10 Amino acid residues selected from Val or Tyr or Y1; X 12 Amino acid residues selected from Ser or Ile or Y1; X 13 Amino acid residues selected from Tyr or Ala or Y1; X 14 Amino acid residues selected from Leu or Nle or Y1; X 15 Amino acid residues selected from Asp or Glu; X 16 Amino acid residues selected from Arg, Glu, Gly, Lys, or Aib, or Y1; X 17 Amino acid residues selected from Glu, Ile, or Gln, or Y1; X 18 Amino acid residues selected from Ala, Aib, or His; X 19an amino acid residue selected from Ala, Aib or Gin; X 20 an amino acid residue selected from Gin, Glu, Lys; X 23 an amino acid residue selected from Ile or Val; X 24 an amino acid residue selected from Ala, Asn or Gin; X 27 an amino acid residue selected from Val or Leu; X 28 an amino acid residue selected from Arg or Ala; X 29 an amino acid residue selected from Gly or Gin; X 30 an amino acid residue selected from Gly, Lys; Y1 is a substituted Lys, Orn, Dap, Dab or Cys residue, in particular carrying a modifying group on the side chain of the Lys, Orn, Dap, Dab or Cys residue. In some embodiments, Y1 is a Lys, Orn, Dap, Dab or Cys residue carrying a substituent on the side chain, which substituent is of the structure; wherein: a is an integer from 1 to 3 (1, 2, 3 can be mentioned); b is 1 or 2; c is an integer from 10 to 30 (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 can be mentioned). a -(y-Glu) b -CO-(CH2) c -COOH; wherein: a is an integer from 1 to 3 (1, 2, 3 can be mentioned); b is 1 or 2; c is an integer from 10 to 30 (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 can be mentioned).

[0022] Some embodiments of the present disclosure provide a GLP-1 analogue of general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is an amino acid residue selected from Tyr; X2 is an amino acid residue selected from Aib; X 10 an amino acid residue selected from Tyr; X 12 an amino acid residue selected from Ile; X 13 an amino acid residue selected from Tyr; X 14 selected from Y1; X 15 an amino acid residue selected from Asp or Glu; X 16 an amino acid residue selected from Arg or Lys; X 17 an amino acid residue selected from Ile; X 18 an amino acid residue selected from Ala; X 19 an amino acid residue selected from Ala; X 20 an amino acid residue selected from Gin; X 23 an amino acid residue selected from Ile or Val; X 24 an amino acid residue selected from Asn; X 27 an amino acid residue selected from Ile or Leu; X28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue having a substituent attached to the side chain of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0023] Some embodiments of the present disclosure provide GLP-1 analogs having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is an amino acid residue selected from Tyr; X2is an amino acid residue selected from Aib; X 10 an amino acid residue selected from Tyr; X 12 an amino acid residue selected from lie; X 13 an amino acid residue selected from Tyr; X 14 selected from Y1; X 15 an amino acid residue selected from Asp or Glu; X 16 an amino acid residue selected from Lys; X 17 an amino acid residue selected from lie; X 18 an amino acid residue selected from Ala; X 19 an amino acid residue selected from Ala; X 20 an amino acid residue selected from Gin; X 23 an amino acid residue selected from Val; X 24 an amino acid residue selected from Asn; X 27 an amino acid residue selected from Leu; X 28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue having a substituent attached to the side chain of the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0024] Some embodiments of the present disclosure provide GLP-1 analogs having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is an amino acid residue selected from Tyr; X2is an amino acid residue selected from Aib or D-Ala; X10 an amino acid residue selected from Y1; X 12 an amino acid residue selected from Ile; X 13 an amino acid residue selected from Tyr; X 14 an amino acid residue selected from Leu or Nle; X 15 an amino acid residue selected from Glu; X 16 an amino acid residue selected from Arg or Lys; X 17 an amino acid residue selected from Ile; X 18 an amino acid residue selected from Ala; X 19 an amino acid residue selected from Ala; X 20 an amino acid residue selected from Gin or Lys; X 23 an amino acid residue selected from Ile or Val; X 24 an amino acid residue selected from Asn or Gin; X 27 an amino acid residue selected from Ile or Leu; X 28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is a Lys, Orn, Dap, Dab or Cys residue bearing a substituent on the side chain, said substituent being a structure of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; wherein: a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0025] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is an amino acid residue selected from Tyr; X2 is an amino acid residue selected from Aib or D-Ala; X 10 an amino acid residue selected from Tyr; X 12 an amino acid residue selected from Y1; X 13 an amino acid residue selected from Tyr; X 14 an amino acid residue selected from Leu or Nle; X 15 an amino acid residue selected from Glu; X 16 an amino acid residue selected from Arg or Lys; X 17 an amino acid residue selected from Ile; X 18 an amino acid residue selected from Ala; X 19 an amino acid residue selected from Ala; X 20 an amino acid residue selected from Gin or Lys; X 23 an amino acid residue selected from Ile or Val; X 24an amino acid residue selected from Asn or Gin; X 27 an amino acid residue selected from lie or Leu; X 28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing on the side chain a substituent of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; wherein: a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0026] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is an amino acid residue selected from Tyr; X2is an amino acid residue selected from Aib or D-Ala; X 10 an amino acid residue selected from Tyr; X 12 an amino acid residue selected from lie; X 13 selected from Y1; X 14 an amino acid residue selected from Leu or Nle; X 15 an amino acid residue selected from Glu; X 16 an amino acid residue selected from Arg or Lys; X 17 an amino acid residue selected from lie; X 18 an amino acid residue selected from Ala; X 19 an amino acid residue selected from Ala; X 20 an amino acid residue selected from Gin or Lys; X 23 an amino acid residue selected from lie or Val; X 24 an amino acid residue selected from Asn or Gin; X 27 an amino acid residue selected from lie or Leu; X 28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing on the side chain a substituent of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; wherein: a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0027] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is an amino acid residue selected from Tyr; X2is an amino acid residue selected from Aib or D-Ala; X3is an amino acid residue selected from Ala or Ser; X4is an amino acid residue selected from Tyr or Phe; X5is an amino acid residue selected from Ile or Val; X6is an amino acid residue selected from Asn or Gln; X7is an amino acid residue selected from Ile or Leu; X8is an amino acid residue selected from Ala or Ser; X9is an amino acid residue selected from Gly; X10is an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing a substituent on the side chain, said substituent being a structure of formula { [2-(2-amino-ethoxy)-ethoxy]-acetyl} 10 is an amino acid residue selected from Tyr; X 12 is an amino acid residue selected from Ile; X 13 is an amino acid residue selected from Tyr; X 14 is Y1; X 15 is an amino acid residue selected from Glu; X 16 is an amino acid residue selected from Arg or Lys; X 17 is an amino acid residue selected from Ile; X 18 is an amino acid residue selected from Ala; X 19 is an amino acid residue selected from Ala; X 20 is an amino acid residue selected from Gln or Lys; X 23 is an amino acid residue selected from Ile or Val; X 24 is an amino acid residue selected from Asn or Gln; X 27 is an amino acid residue selected from Ile or Leu; X 28 is an amino acid residue selected from Ala; X 29 is an amino acid residue selected from Gly; X 30 is an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing a substituent on the side chain, said substituent being a structure of formula { [2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; wherein: a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0028] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is an amino acid residue selected from Tyr; X2is an amino acid residue selected from Aib; X3is an amino acid residue selected from Ala or Ser; X4is an amino acid residue selected from Tyr or Phe; X5is an amino acid residue selected from Ile or Val; X6is an amino acid residue selected from Asn or Gln; X7is an amino acid residue selected from Ile or Leu; X8is an amino acid residue selected from Ala or Ser; X9is an amino acid residue selected from Gly; X10is an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing a substituent on the side chain, said substituent being a structure of formula { [2-(2-amino-ethoxy)-ethoxy]-acetyl} 10 is an amino acid residue selected from Tyr; X 12 is an amino acid residue selected from Ile; X 13 is an amino acid residue selected from Tyr; X 14 is Y1; X 15 is an amino acid residue selected from Glu; X 16 is an amino acid residue selected from Arg or Lys; X 17 is an amino acid residue selected from Ile; X 18 is an amino acid residue selected from Ala; X 19 is an amino acid residue selected from Ala; X20 an amino acid residue selected from Gin; X 23 an amino acid residue selected from lie or Val; X 24 an amino acid residue selected from Asn; X 27 an amino acid residue selected from lie or Leu; X 28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing on the side chain a substituent of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0029] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is an amino acid residue selected from Tyr; X2is an amino acid residue selected from Aib or D-Ala; X 10 an amino acid residue selected from Tyr; X 12 an amino acid residue selected from lie; X 13 an amino acid residue selected from Tyr; X 14 an amino acid residue selected from Leu or Nle; X 15 an amino acid residue selected from Glu; X 16 an amino acid residue selected from Y1; X 17 an amino acid residue selected from lie; X 18 an amino acid residue selected from Ala; X 19 an amino acid residue selected from Ala; X 20 an amino acid residue selected from Gin or Lys; X 23 an amino acid residue selected from lie or Val; X 24 an amino acid residue selected from Asn or Gin; X 27 an amino acid residue selected from lie or Leu; X 28 an amino acid residue selected from Ala; X 29 an amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing on the side chain a substituent of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c-COOH; wherein: a is an integer from 1 to 3; b is 1 or 2; and c is an integer from 10 to 30.

[0030] Some embodiments of the present disclosure also provide GLP-1 analogs having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1is selected from the group consisting of an amino acid residue of Tyr; X2is selected from the group consisting of an amino acid residue of Aib or D-Ala; X 10 is selected from the group consisting of an amino acid residue of Tyr; X 12 is selected from the group consisting of an amino acid residue of Ile; X 13 is selected from the group consisting of an amino acid residue of Tyr; X 14 is selected from the group consisting of an amino acid residue of Leu or Nle; X 15 is selected from the group consisting of an amino acid residue of Glu; X 16 is selected from the group consisting of an amino acid residue of Arg or Lys; X 17 is selected from the group consisting of Y1; X 18 is selected from the group consisting of an amino acid residue of Ala; X 19 is selected from the group consisting of an amino acid residue of Ala; X 20 is selected from the group consisting of an amino acid residue of Gin or Lys; X 23 is selected from the group consisting of an amino acid residue of Ile or Val; X 24 is selected from the group consisting of an amino acid residue of Asn or Gin; X 27 is selected from the group consisting of an amino acid residue of Ile or Leu; X 28 is selected from the group consisting of an amino acid residue of Ala; X 29 is selected from the group consisting of an amino acid residue of Gly; X 30 is selected from the group consisting of an amino acid residue of Gly; Y1is a Lys, Orn, Dap, Dab or Cys residue bearing a substituent on the side chain, said substituent being a group of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; wherein: a is an integer from 1 to 3; b is 1 or 2; and c is an integer from 10 to 30.

[0031] Some embodiments of the present disclosure also provide GLP-1 analogs having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X 10 , X 12 , X 13 , X 14 , X 16 and X 17 are each independently selected from Y1; wherein Y1is a Lys, Orn, Dap, Dab or Cys residue bearing a substituent on the side chain, said substituent being a group of formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b-CO-(CH2) c -COOH; a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

[0032] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein a is 2, b is 1 or 2, and c is an integer from 16 to 20 (16, 17, 18, 19, 20 can be mentioned).

[0033] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein a is 2, b is 1 or 2, and c is 16, 18 or 20.

[0034] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, X 10 is Y1; Y1 is Lys bearing in the side chain a substituent of formula {[2-(2-amino- ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

[0035] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, X 12 is Y1; Y1 is Lys bearing in the side chain a substituent of formula {[2-(2-amino- ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

[0036] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, X 13 is Y1; Y1 is Lys bearing in the side chain a substituent of formula {[2-(2-amino- ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

[0037] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, X 14Y1; Y1 is Lys with a substituent in the side chain, which is a structure represented by formula { [2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

[0038] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, X 16 Y1; Y1 is Lys with a substituent in the side chain, which is a structure represented by formula { [2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

[0039] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, X 17 Y1; Y1 is Lys with a substituent in the side chain, which is a structure represented by formula { [2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH2) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

[0040] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, the side chain amino group of the Lys residue in Y1 is covalently linked to the substituent by forming an amide bond.

[0041] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the following structure:

[0042]

[0043] K(-OEG-OEG-yGlu-C20-OH) has the following structure:

[0044]

[0045] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein:

[0046] K(-OEG-OEG-yGlu-C18-OH) has the following structure:

[0047]

[0048] K(-OEG-OEG-yGlu-C20-OH) has the following structure:

[0049]

[0050] Some embodiments of the present disclosure also provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein:

[0051] Some embodiments of the present disclosure provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof, wherein X1 is selected from the amino acid residue of Tyr; X2 is selected from the amino acid residue of Aib; X 10 selected from the amino acid residue of Tyr; X 12 selected from the amino acid residue of Ile; X 13 selected from the amino acid residue of Tyr; X 14 selected from Y1; X 15 selected from the amino acid residue of Asp or Glu; X 16 selected from the amino acid residue of Arg or Lys; X 17 selected from the amino acid residue of Ile; X 18 selected from the amino acid residue of Ala; X 19 selected from the amino acid residue of Ala; X 20 selected from the amino acid residue of Gln; X 23 selected from the amino acid residue of Ile or Val; X 24 selected from the amino acid residue of Asn; X 27 selected from the amino acid residue of Ile or Leu; X 28 selected from the amino acid residue of Ala; X 29 selected from the amino acid residue of Gly; X 30 selected from the amino acid residue of Gly; Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the following structure:

[0052]

[0053] K(-OEG-OEG-yGlu-C20-OH) has the following structure:

[0054]

[0055] Some embodiments of this disclosure provide GLP-1 analogs having general formula (I) or pharmaceutically acceptable salts thereof, wherein X1 is selected from amino acid residues of Tyr; X2 is selected from amino acid residues of Aib; X 10 amino acid residues selected from Tyr; X 12 Selected amino acid residues from Ile; X 13 amino acid residues selected from Tyr; X 14 Selected from Y1; X 15 Amino acid residues selected from Asp or Glu; X 16 Amino acid residues selected from Arg or Lys; X 17 Selected from amino acid residues of Ile; X 18 Selected from amino acid residues of Ala; X 19 Selected from amino acid residues of Ala; X 20 Amino acid residues selected from Gln; X 23 Amino acid residues selected from Ile or Val; X 24 Amino acid residues selected from Asn; X 27 Amino acid residues selected from Ile or Leu; X 28 Selected from amino acid residues of Ala; X 29 Selected from amino acid residues of Gly; X 30 The amino acid residues are selected from Gly; Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the following structure:

[0056]

[0057] K(-OEG-OEG-yGlu-C20-OH) has the following structure:

[0058]

[0059] Some embodiments of this disclosure also provide GLP-1 analogs having general formula (I) or pharmaceutically acceptable salts thereof, wherein the GLP-1 analog is shown in general formula (II) (SEQ ID NO:20): H-YAibEGTFTSDYSIYX 14 X 15X 16 IAAQEFX 23 NWLX 27 AGGPSSGAPPPS-NH2 (II), wherein X 14 is K or L, X 15 is D or E, X 16 is K or R, X 23 is V or I, X 27 is I or L.

[0060] Some embodiments of the present disclosure further provide a GLP-1 analog having the general formula (I) or a pharmaceutically acceptable salt thereof, the GLP-1 analog is selected from the group consisting of compounds shown in the following numbers 1-18:

[0061]

[0062]

[0063] Some embodiments of the present disclosure further provide a GLP-1 analog having the general formula (I) or a pharmaceutically acceptable salt thereof, the GLP-1 analog is selected from the group consisting of compounds shown in the following numbers 1-18:

[0064]

[0065] In some embodiments, the GLP-1 analog of the present disclosure is selected from the group consisting of compounds shown in 7#, 12#, 13#, 14#, 15#, 16#, 17#, or 18# in Figure 3.

[0066] Some embodiments of the present disclosure further provide a pharmaceutical composition comprising:

[0067] 1) a GLP-1 analog having the general formula (I) or a pharmaceutically acceptable salt thereof, and

[0068] 2) a pharmaceutically acceptable excipient or a pharmaceutical carrier.

[0069] In some embodiments, the pharmaceutical composition can contain 0.01 to 99% by weight of the GLP-1 analog, or the amount of the GLP-1 analog in a unit dose of the pharmaceutical composition is 0.1-2000 mg, in some embodiments, 1-1000 mg.

[0070] Some embodiments of the present disclosure further provide a GLP-1 analog having the general formula (I) or a pharmaceutically acceptable salt thereof, and a use of the pharmaceutical composition comprising the same in the preparation of a medicament for treating non-insulin dependent diabetes mellitus / Type II diabetes mellitus, insulin dependent diabetes mellitus, obesity, non-alcoholic fatty liver, hepatic steatosis, dyslipidemia associated with insulin resistance, and / or diabetic dyslipidemia.

[0071] Some embodiments of the present disclosure provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament.

[0072] Some embodiments of the present disclosure provide a GLP-1 analogue having the general formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament for treating non-insulin dependent diabetes mellitus / Type II diabetes, insulin dependent diabetes mellitus, obesity, non-alcoholic fatty liver, hepatic steatosis, dyslipidemia associated with insulin resistance, and / or diabetic dyslipidemia.

[0073] Some embodiments of the present disclosure provide a method for treating non-insulin dependent diabetes mellitus / Type II diabetes, insulin dependent diabetes mellitus, obesity, non-alcoholic fatty liver, hepatic steatosis, dyslipidemia associated with insulin resistance, and / or diabetic dyslipidemia, comprising administering to a subject in need thereof a GLP-1 analogue having the general formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0074] The present disclosure provides compounds that can simultaneously activate GLP-1 receptor and GIP receptor, in some embodiments, the GLP-1 analogue has a higher agonist activity on GLP-1R than on GIP receptor.

[0075] In some embodiments, the ratio of agonist activity of a GLP-1 analogue of the present disclosure at GLP-1R to agonist activity at GIP receptor is (1-10):1, (1.1-10):1, (1.1-9.5):1, (1.1-9):1, (1.1-8.5):1, (1.1-8):1, (1.1-7.5):1, (1.1-7):1, (1.1-6.5):1, (1.1-6):1, (1.2-10):1, (1.2-9.5):1, (1.2-9):1, (1.2-8.5):1, (1.2-8):1, (1.2-7.5):1, (1.2-7):1, (1.2-6.5):1, (1.2-6):1, (1.3-10):1, (1.3-9.5):1, (1.3-9):1, (1.3-8.5):1, (1.3-8):1, (1.3-7.5):1, (1.3-7):1, (1.3-6.5):1, (1.3-6):1, (1.4-10):1, (1.4-9.5):1, (1.4-9):1, (1.4-8.5):1, (1.4-8):1, (1.4-7.5):1, (1.4-7):1, (1.4-6.5):1, (1.4-6):1, (1.5-10):1, (1.5-9.5):1, (1.5-9):1, (1.5-8.5):1, (1.5-8):1, (1.5-7.5):1, (1.5-7):1, (1.5-6.5):1, (1.5-6):1, (2-10):1, (2-9.5):1, (2-9):1, (2-8.5):1, (2-8):1, (2-7.5):1, (2-7):1, (2-6.5):1, (2-6):1, (2.5-10):1, (2.5-9.5):1, (2.5-9):1, (2.5-8.5):1, (2.5-8):1, (2.5-7.5):1, (2.5-7):1, (2.5-6.5):1, (2.5-6):1, (3-10):1, (3-9.5):1, (3-9):1, (3-8.5):1, (3-8):1, (3-7.5):1, (3-7):1, (3-6.5):1, (3-6):1, (3.5-10):1, (3.5-9.5):1, (3.5-9):1, (3.5-8.5):1, (3.5-8):1, (3.5-7.5):1, (3.5-7):1, (3.5-6.5):1, (3.5-6):1, (4-10):1, (4-9.5):1, (4-9):1, (4-8.5):1, (4-8):1, (4-7.5):1, (4-7):1, (4-6.5):1, (4-6):1, (4.5-10): 1, (4.5-9.5): 1, (4.5-9): 1, (4.5-8.5): 1, (4.5-8): 1, (4.5-7.5): 1, (4.5-7): 1, (4.5-6.5): 1, (4.5-6): 1, (5-10): 1, (5-9.5): 1, (5-9): 1, (5-8.5): 1, (5-8): 1, (5-7.5): 1, (5-7): 1, (5-6.5): 1, (5-6): 1, (5-5.5): 1, (5.1-5.5): 1, (5.2-5.4): 1, (5.2-5.3): 1, or any numerical range or value point in between, for example, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 2:1, about 2.5:1, about 3:1, about 3.5:1, about 4:1, about 4.5:1, about 5:1, about 5.2:1, about 5.3:1, about 5.4:1, about 5.5:1, about 6:1, about 6.5:1, about 7:1, about 7.5:1, about 8:1, about 8.5:1, about 9:1, about 9.5:1, about 10:1. The above ratios are normalized from data from corresponding agonist activity in vitro assays. For example, the corresponding agonist activity can be determined by a cAMP-Gs Kinetics Assay Kit. In the context, the expression (1-10): 1 and the expression 1:1 to 10:1 have the same meaning.

[0076] In another embodiment, the present disclosure provides the above GLP-1 analogs and pharmaceutically acceptable salts thereof. The GLP-1 analogs provided by the present disclosure belong to amphoteric compounds, which can exhibit both acidic and basic properties. Those skilled in the art can use an acidic or basic compound to react with the GLP-1 analogs provided by the present disclosure to form a salt by using known techniques.

[0077] The pharmaceutical compositions containing the GLP-1 analogs according to the present disclosure can be used for treating patients in need of such treatment by parenteral administration. The parenteral administration route can be selected from subcutaneous injection, intramuscular injection, or intravenous injection. The polypeptide dual agonist compounds of the present disclosure can also be administered by transdermal route, which can be selected from iontophoresis patch; or transmucosal route.

[0078] The GLP-1 analogs provided herein are synthesized by solid phase synthesis. As an example, the synthetic support is Rink-amide MBHA (Xi'an Blue Sky Science and Technology) resin. During the synthesis, the alpha-amino group of the amino acid derivative used is protected by a Fmoc group (fluorenylmethyloxycarbonyl). As an example, for the side chain of the amino acid, the following protecting groups are selected according to the functional group: the thiol group of the cysteine side chain, the amino group of the asparagine and glutamine side chain, the imidazole group of the histidine side chain are protected by Trt (trityl); the guanidino group of the arginine side chain is protected by Pbf (2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl); the indole group of the tryptophan side chain, the amino group of the lysine side chain are protected by Boc (tert-butyloxycarbonyl); the carboxyl group of the aspartic acid and glutamic acid side chain, the hydroxyl group of the threonine side chain, the phenol group of the tyrosine side chain, the hydroxyl group of the serine side chain are protected by t-Bu (tert-butyl). As an example, during the synthesis, first the carboxyl group of the C-terminal amino acid residue of the polypeptide is condensed to the high molecular insoluble Rink-amide MBHA resin in the form of an amide bond; then the Fmoc protecting group on the alpha-amino group is removed with a 20% 4-methylpiperidine solution in nitrogen, nitrogen-dimethylformamide (DMF); then in excess, the solid phase support is condensed with the next amino acid derivative in the polypeptide sequence to form an amide bond to extend the peptide chain. The operation of "condensation → washing → deprotection → washing → next round of amino acid condensation" is repeated to achieve the length of the polypeptide to be synthesized; finally, the polypeptide is cleaved from the solid phase support by reacting with a mixed solution of trifluoroacetic acid: water: triisopropylsilane (as an example, 90:5:5, v:v:v); then precipitated by 5 volumes of cold methyl tert-butyl ether to obtain a solid crude product of the GLP-1 analog. The solid crude product of the polypeptide is dissolved in a mixed solution of acetonitrile / water containing 0.1% trifluoroacetic acid, and then purified and separated by C-18 reverse phase preparative chromatography column to obtain a pure product of the GLP-1 analog.

[0079] According to some embodiments, the present disclosure also provides a kit-of-parts comprising:

[0080] - a GLP-1 analog or a pharmaceutically acceptable salt thereof according to the present disclosure;

[0081] - another therapeutic agent selected from any one or a combination of the following: an anti-obesity agent, an anti-diabetic agent, an anti-hypertensive agent, a lipid-lowering agent. In some embodiments, the GLP-1 analog or a pharmaceutically acceptable salt thereof and the other therapeutic agent are separately located in different containers. In some embodiments, the GLP-1 analog or a pharmaceutically acceptable salt thereof and the other therapeutic agent are administered to a subject separately or in combination (e.g., simultaneously or sequentially).

[0082] In certain embodiments, the pharmaceutical compositions of the present disclosure and an administration device (e.g., and a syringe, an injection pen, or an auto-injector) are provided in combination. As one example, the pharmaceutical compositions of the present disclosure are pre-filled in the administration device to facilitate self-administration by a subject at home. As another example, the pharmaceutical compositions of the present disclosure and the administration device are provided separately. BRIEF DESCRIPTION OF DRAWINGS

[0083] Figure 1 shows the effect of the compounds of the present disclosure on the rate of change in body weight in diet-induced obese mice.

[0084] Figure 2 shows the effect of the compounds of the present disclosure on daily food intake in diet-induced obese mice.

[0085] Figure 3 shows the structure of exemplary compounds of the present disclosure. DETAILED DESCRIPTION

[0086] For the purposes of the present disclosure, certain technical and scientific terms are specifically defined below. Unless specifically defined herein, all other technical and scientific terms used in the present disclosure have the meanings that are commonly understood by one of ordinary skill in the art in the field of the present disclosure.

[0087] The amino acid sequences of the present disclosure contain the standard one-letter or three-letter codes for the twenty amino acids, and all amino acid residues in the present disclosure are in the preferred L-form, unless explicitly stated otherwise. In addition, Aib is alpha aminoisobutyric acid, D-Ala is D-form alanine, Orn is ornithine, Dap is 2,3-diaminopropionic acid, and Dab is 2,4-diaminobutyric acid.

[0088] The term "agonist" is defined as a substance that has an activating effect on the GLP-1 receptor or on the GIP receptor.

[0089] The term "GLP-1 / GIP dual agonist" as used in the context of the present disclosure refers to a substance or ligand that can activate both the GLP-1 receptor and the GIP receptor.

[0090] In the present disclosure, the term "treatment" includes inhibiting, slowing, stopping, or reversing the progression or severity of an existing symptom or condition.

[0091] "Natural amino acids" refer to the 20 conventional amino acids, namely alanine (A), cysteine (C), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), proline (P), glutamine (Q), arginine (R), serine (S), threonine (T), valine (V), tryptophan (W), and tyrosine (Y).

[0092] A "non-natural amino acid" refers to an amino acid that is not naturally encoded, or not found in the genetic code of any organism. For example, a non-natural amino acid can be a purely synthetic compound. Examples of non-natural amino acids include, but are not limited to, hydroxyproline, gamma-carboxyglutamate, O-phosphoserine, azetidinecarboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid, beta-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminohexanoic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminoheptanedioic acid, t-butylglycine, 2,4-diaminoisobutyric acid (Dap), desmosine, 2,2'-diaminopimelic acid, 2,3-diaminopropionic acid (Dab), N-ethylglycine, N-methylglycine, N-ethylasparagine, homoproline, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylalanine, N-methylglycine, N-methylisoleucine, N-methylpentylglycine, N-methylvaline, naphthalanine, norvaline, norleucine, ornithine (Orn), D-ornithine, D-arginine, p-aminophenylalanine, pentylglycine, pipecolic acid, and thioproline. In addition, the term also includes derivatives resulting from chemical modifications at the C-terminal carboxyl group (or N-terminal amino group and / or side chain functional groups) of a natural amino acid (or non-natural amino acid).

[0093] The term "alkyl" refers to saturated aliphatic hydrocarbon groups, which are straight-chain or branched-chain groups containing 1 to 20 carbon atoms, such as alkyl groups containing 1 to 8 carbon atoms, such as alkyl groups containing 1 to 6 carbon atoms, such as alkyl groups containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof, and the like. Alkyl groups may, for example, be lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples including methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. Alkyl groups can be substituted or unsubstituted, when substituted, the substituents can be substituted at any accessible attachment point, which can be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halo, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylate. Substituted alkyl groups of the present disclosure can be methyl, ethyl, i-propyl, t-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl, or hydroxyl-substituted alkyl.

[0094] The different expressions "X is selected from A, B, or C", "X is selected from A, B and C", "X is A, B or C", "X is A, B and C" and the like in the present disclosure express the same meaning, i.e. X can be any one or several of A, B, C.

[0095] The "modification" of an amino acid according to the present disclosure refers to a substitution, addition or deletion of an amino acid, including the substitution or addition of any one or more of the 20 natural amino acids.

[0096] The term "native GLP-1" refers to a naturally occurring molecule of the glucagon family of peptides or the exendin family of peptides, wherein: the glucagon family of peptides is encoded by the preproglucagon gene and includes three small peptides with high homology, i.e. glucagon (1-29), GLP-1 (1-37) and GLP-2 (1-33); the exendin is a peptide expressed in lizards and, like GLP-1, is insulinotropic. In some embodiments, the term "native GLP-1" also refers to human GLP-1 (7-37) and human GLP-1 (7-36).

[0097] The term "GLP-1 analogue" refers to a modification of up to 25, up to 24, up to 23, up to 22, up to 21, up to 20, up to 19, up to 18, up to 17, up to 16, up to 15, up to 14, up to 13, up to 12, up to 11, up to 10, up to 9, up to 8, up to 7, up to 6, up to 5, up to 4, up to 3, up to 2, or 1 amino acid modification or chemical modification compared to native GLP-1, in particular compared to human GLP-1 (7-37) and human GLP-1 (7-36), which can be an amino acid substitution, addition and / or deletion; which can be a chemical modification selected from the group consisting of amide, carbohydrate, alkyl, acyl, ester, polyethylene glycol (PEG) group, sialylation group, glycosylation group and the like.

[0098] The "substitution" of an amino acid according to the present disclosure refers to the substitution of one amino acid residue by a different amino acid residue.

[0099] The term "polyethylene glycol" or "PEG" refers to a mixture of polycondensates of ethylene oxide and water, present in linear or branched form, of the general formula H(OCH2CH2) n OH, wherein n is minimally equal to 9. If not further specified, this term includes polymers of polyethylene glycol having an average total molecular weight selected from the range of 5,000 to 40,000 Dalton.

[0100] The term "fatty acid" refers to a carboxylic acid having a long fatty acid family tail (chain) which can be saturated or unsaturated; a fatty acid in the present disclosure is a carboxylic acid having a C4-C30 straight chain or branched chain aliphatic group.

[0101] The term "peptide" as used in the present disclosure encompasses the scope of peptides having modified amino and carboxyl termini. For example, an amino acid chain comprising a terminal carboxylic acid replaced with an amide group is also included within the designation of an amino acid sequence of a natural amino acid.

[0102] Any hydrogen atom described in the present disclosure can be replaced by its isotope (protium, deuterium, tritium), and any hydrogen atom in the compounds of the present disclosure described herein can also be replaced by an isotope atom.

[0103] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and thus the description includes instances where the event or circumstance occurs and instances where it does not. For example, "heterocyclic group optionally substituted with alkyl" means that alkyl can or can not be present, and the description includes instances where the heterocyclic group is substituted with alkyl and instances where the heterocyclic group is not substituted with alkyl.

[0104] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3, of a group are independently of each other replaced with a substituent. Substituents are only in their possible chemical positions, which can or cannot be possible (experimentally or theoretically) to be determined by one skilled in the art without undue effort. For example, an amino group with a free hydrogen can not be stable when bound to a carbon atom with an unsaturated (e.g., olefinic) bond.

[0105] "Pharmaceutical composition" means a mixture of one or more of the compounds described herein, or physiologically / pharmaceutically acceptable salts or prodrugs thereof, with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to a subject, and to facilitate absorption, thereby facilitating biological activity.

[0106] "Agonist activity" means the ability of a compound according to the present disclosure to activate the human GIP receptor and the human GLP-1 receptor. In some examples, "agonist activity" is expressed in terms of relative activity, and in particular refers to the ratio of the ability of a compound of the present disclosure to activate the GLP-1 R to the ability to activate the GIP receptor.

[0107] "Pharmaceutically acceptable salt" means a salt of a compound of the present disclosure that is safe and effective for use in mammals and possesses the desired biological activity.

[0108] Semaglutide refers to a once-weekly GLP-1 receptor single agonist polypeptide drug developed by Novo Nordisk, which has been approved and marketed in the United States, Japan and the European Union.

[0109] LY3298176 refers to a once-weekly GIP receptor / GLP-1 receptor dual agonist polypeptide drug developed by Eli Lilly and Company, which is currently in phase III clinical trials in multiple countries. The structure is as follows:

[0110] YAibEGTFTSDYSIAibLDKIAQKAFVQWLIAGGPSSGAPPPS-NH2, wherein the fatty acid is modified on K at position 20.

[0111] Examples

[0112] In order to explain the present disclosure in more detail, the following specific embodiments are provided in the specification, but the scope of the present disclosure is not limited to only this. The experimental methods not specified in the specific conditions in the examples of the present disclosure are generally according to the conventional conditions; or according to the conditions suggested by the manufacturer of the raw materials or commodities. The reagents not specified in the specific source are the conventional reagents purchased on the market.

[0113] 1. Experimental reagents

[0114] Table 1. Reagents used and sources

[0115]

[0116]

[0117] 2. Experimental instruments

[0118] Table 2. Instruments used and sources

[0119] No. Instrument Source 1 H-CLASS analytical ultra-performance liquid chromatograph WATERS 2 Agilent 1290-6530 ultra-performance liquid chromatograph / mass spectrometer Agilent 3 Labconco multifunctional freeze dryer Thermo-Fisher Scientific 4 Prep150 preparative high-performance liquid chromatograph WATERS 5 Prelude-X polypeptide automatic synthesizer Protein Technology Inc 6 Multichannel high-speed centrifuge Sigma

[0120] Example 1 Chemical synthesis of compound 18#

[0121] 1. Synthesis of polypeptide backbone

[0122] Rink-amide MBHA resin (degree of substitution: 0.48 mmole / g, 0.1 mmol) was weighed into a polypropylene polypeptide synthesis solid phase reaction tube; the resin was swelled by adding N,N-dimethylformamide (DMF, 10 mL) under nitrogen-blowing for 10 minutes; the DMF was removed by vacuum suction, and the resin was washed with DMF (10 mL); after the resin was washed twice, polypeptide solid phase synthesis was performed on a Prelude-X polypeptide automatic synthesizer using the Fmoc / tBu strategy, in which 10 equivalents of amino acid residues activated by HCTU and 4-methylmorpholine (molar ratio of HCTU, 4-methylmorpholine, amino acid residues: 1:2:1) were used for amide bond condensation in DMF at room temperature for 25 minutes to achieve coupling. Deprotection of the N-terminal Fmoc protecting group was performed by using a 20% 4-methylpiperidine DMF solution at room temperature for 2 times (10 minutes each time). In the polypeptide backbone synthesis, Boc-L-Tyr(tBu)-OH was selected for the N-terminal amino acid residue to construct and perform secondary condensation, which is necessary to improve the quality of the crude peptide.

[0123] 2. Selective deprotection of the resin peptide protecting group Mtt and modification of the side chain fatty acid

[0124] After the extension of the polypeptide backbone (or resin peptide) described above was completed, a mixed solution of 30% hexafluoroisopropanol in dichloromethane (10 mL) was added, and after oscillation reaction at room temperature for 45 minutes, the mixed solution was removed; a mixed solution of 30% hexafluoroisopropanol in dichloromethane (10 mL) was added again, and after oscillation reaction at room temperature for 45 minutes, the mixed solution was removed. After the reaction was completed, the resin was washed with DMF 6 times. The Prelude-X automatic polypeptide synthesizer was used to extend the side chain of the 14th lysine, and the additional coupling / deprotection cycle involved the amino acid components Fmoc-NH-PEG2-COOH and Fmoc-L-Glu-OtBu. In all couplings, 10 equivalents of amino acid residues activated by HCTU and 4-methylmorpholine (molar ratio of HCTU, 4-methylmorpholine, amino acid residues: 1:2:1) were used for coupling in DMF at room temperature for 25 minutes. Deprotection of the N-terminal Fmoc protecting group was performed by using a 20% 4-methylpiperidine DMF solution at room temperature for 2 times, 10 minutes each time. After the final resin was washed with DCM and DMF three times, respectively, a mixed solution of 10 equivalents of HOOC-(CH2) 18 -COOtBu, 10 equivalents of HCTU and 20 equivalents of diisopropylethylamine (DIEA) in DMF (8 mL) was added, and the reaction was carried out at room temperature for 4 hours to complete the modification of the side chain fatty acid.

[0125] 3. Product cleavage

[0126] The resin peptide obtained in the previous step was washed with DMF and DCM for 3 times, respectively, and then dried in vacuum. Freshly prepared cleavage solution (trifluoroacetic acid: triisopropylsilane: water = 90:5:5, by volume) was added and the reaction was allowed to proceed at room temperature for 3-4 hours. After the reaction was completed, the resin was filtered and washed with trifluoroacetic acid for 2 times. The filtrate was combined and a large amount of chilled methyl tert-butyl ether was added to precipitate the solid. After centrifugation, the supernatant was removed and the crude polypeptide of compound 18# was obtained.

[0127] 4. Purification by reverse phase liquid chromatography

[0128] The crude polypeptide of compound 18# was dissolved in a mixed solvent containing 0.1% trifluoroacetic acid, 20% acetonitrile, and 20% acetic acid / water, and filtered through a 0.22 μm membrane. Separation was performed using a WATERS Prep 150 LC reverse phase high performance liquid chromatography system, with buffer A (0.1% trifluoroacetic acid, 10% acetonitrile, aqueous solution) and buffer B (0.1% trifluoroacetic acid, 90% acetonitrile, aqueous solution). The chromatographic column was an X-SELECT OBD C-18 reverse phase chromatographic column, and the detection wavelength of the chromatograph was set to 220 nm during the purification process, with a flow rate of 15 mL / min. The product-related fractions were collected and lyophilized to obtain the pure polypeptide of compound 1#, with a yield of 18%. The purity of the polypeptide was determined by analytical high performance liquid chromatography and ultra-high performance liquid chromatography / mass spectrometry, and the purity was 92.81%. The molecular structure of compound 18# is H-YAibEGTFTSDYSIYK(OEG-OEG-yGlu-C20-OH)EKIAAQEFVNWLLAGGPSSGAPPPS-NH2, as shown in the structure of 18# in Figure 3.

[0129] Example 2. Chemical synthesis of other compounds

[0130] The compounds in Table 3 were synthesized using the experimental protocol of Example 1:

[0131] Table 3. Compounds of the present disclosure

[0132]

[0133] The purity of the compounds was determined by analytical high performance liquid chromatography and ultra-high performance liquid chromatography / mass spectrometry, and the purity of some of the compounds is shown in Table 4 below:

[0134] Table 4. Purity and molecular weight of compounds 8#-11# determined by the combined use of analytical high performance liquid chromatography and liquid chromatography / mass spectrometry

[0135] Compound number Purity 8# 96.30% 9# 93.28% 10# 94.56% 11# 92.18%

[0136] Biological test evaluation

[0137] The present disclosure is further described in connection with the following test examples, which are not meant to limit the scope of the present disclosure.

[0138] 1. Experimental reagents

[0139] Table 5. Reagents used in this experiment and sources

[0140]

[0141] 2. Experimental instruments

[0142] Table 6. Instruments used in this experiment and sources

[0143]

[0144]

[0145] Example 3 Evaluation of agonist activity of compounds of the present disclosure on glucagon-like peptide-1 receptor (GLP-1R)

[0146] 1. Purpose of the experiment:

[0147] The purpose of this test example is to measure the agonist activity of compounds of the present disclosure on glucagon-like peptide-1 receptor (GLP-1R).

[0148] 2. Experimental method:

[0149] The frozen CHO-K1 / GLP-1R / CRE-luc stable cell line (which can be prepared by conventional methods in the art) was taken out from the liquid nitrogen tank, placed in a 37°C water bath for rapid thawing, resuspended with DMEM / F12 medium (Gibco Cat#11330032), washed the cells once after centrifugation, resuspended with experimental buffer, i.e. DMEM / F12 medium containing 0.1% casein (Sigma Cat#C3400), adjusted the cell density with experimental buffer, and plated in a 384-well plate (Sigma Cat#CLS4514) at a density of 2500 cells / 5 μL / well, then added 2.5 μL of IBMX working solution (Sigma Cat#I7018) prepared with buffer to each well, with a final concentration of 0.5 mM, and 2.5 μL of gradient-diluted polypeptide sample, centrifuged at 1000 rpm for 1 minute, mixed well by shaking for 30 seconds, and incubated at room temperature for 30 minutes. Detection was performed using Cisbio cAMP-Gs Dynamic kit (Cisbio Cat#62AM4PEC), and cAMP-d2 and Anti-cAMP-Eu 3+- Cryptate was diluted 20 times with cAMP Lysis & Detection Buffer and mixed well. 5 μL of diluted cAMP-d2 solution was added to each well, followed by 5 μL of diluted Anti-cAMP-Eu 3+ - Cryptate solution was mixed well by shaking for 30 seconds and incubated at room temperature for 1 hour in the dark.

[0150] 3. Experimental data processing method:

[0151] Biotek Synergy H1 microplate reader was used to read the HTRF signal, with excitation wavelength of 320 nm and emission wavelength of 620 nm and 665 nm. The signal ratio (665 nm / 620 nm*10,000) was calculated, and the signal ratio and sample concentration were non-linearly fitted using a four-parameter equation in GraphPad Prism 6 to obtain the EC 50 value. The specific data are shown in Table 7 below.

[0152] Example 4 Evaluation of the agonist activity of the compound of the present disclosure on glucose-dependent insulinotropic peptide receptor (GIP receptor)

[0153] 1. Experimental purpose:

[0154] The agonist activity of the test compound on glucose-dependent insulinotropic peptide receptor (GIP receptor) was tested.

[0155] 2. Experimental method:

[0156] CHO-K1 cells were collected and adjusted to the appropriate density, plated in 6-well plates at 2 mL per well, and incubated overnight at 37 °C in a 5% CO2 incubator. Transfection mix (hGIP receptor plasmid, Fugene HD (Promega Cat# E2311), OptiMEM (Gibco Cat# 31985070) was mixed and allowed to stand at room temperature for 15 minutes. The transfection mix was added to the corresponding cell wells at a volume of 100 μL. The CHO-K1 cells were allowed to overexpress the hGIP receptor on the cell surface for 24 hours. After the transfection was complete, the cells in the 6-well plates were collected and washed once with experimental buffer, DMEM / F12 medium (Gibco Cat# 11330032) containing 0.1% casein (Sigma Cat# C3400). The cells were adjusted to the appropriate density using experimental buffer and plated in a 384-well plate (Sigma Cat# CLS4514) at a density of 5000 cells / 5 μL / well. Then 2.5 μL of IBMX working solution (Sigma Cat# I7018) prepared in buffer was added to each well, and the final concentration of IBMX was 0.5 mM. Then 2.5 μL of the gradient-diluted polypeptide sample was added to each well. The plate was centrifuged at 1000 rpm for 1 minute, mixed by shaking for 30 seconds, and incubated at room temperature for 30 minutes. The Cisbio cAMP-Gs Dynamic kit (Cisbio Cat# 62AM4PEC) was used for detection. The cAMP-d2 and Anti-cAMP-Eu3+-Cryptate were mixed by diluting 20 times with cAMP Lysis & Detection Buffer. Then 5 μL of the diluted cAMP-d2 solution and 5 μL of the diluted Anti-cAMP-Eu3+-Cryptate solution were added to each well, mixed by shaking for 30 seconds, and incubated at room temperature for 1 hour in the dark.

[0157] 3. Experimental data processing method:

[0158] The Biotek Synergy H1 microplate reader was used to read the HTRF signal, with an excitation wavelength of 320 nm and emission wavelengths of 620 nm and 665 nm. The signal ratio (665 nm / 620 nm*10,000) was calculated, and the signal ratio and sample concentration were non-linearly fitted using a four-parameter equation in GraphPad Prism 6 to obtain the EC 50 value. The specific values are shown in Tables 7 and 8 below.

[0159] Table 7. Agonist activity test results of human GLP-1R and human GIP receptor

[0160]

[0161] Table 8. Results of agonist activity test of human GLP-1R and human GIP receptor

[0162]

[0163] 4. Experimental Conclusion:

[0164] By designing the polypeptide skeleton and subsequent site-specific modification of fatty acids, the compounds of the present disclosure have stronger GLP-1 / GIP receptor agonist activity compared to many GLP-1 / GIP receptor dual agonist polypeptides in the art, and thus have better potential for treating metabolic diseases. In addition, LY3298176 shows preferential activity on GIP receptor, unlike the compounds 12#-18# of the present disclosure, which show preferential activity on GLP-1R.

[0165] Example 5. Stability test of some compounds of the present disclosure

[0166] Stability in plasma is important for therapeutic polypeptide drugs, because polypeptide drugs are likely to be sensitive to polypeptide hydrolases and proteolytic enzymes in plasma. Polypeptides that are unstable in plasma will have their half-lives and efficacy affected.

[0167] 1. Experimental Purpose:

[0168] The purpose of this experiment is to test the stability of some compounds of the present disclosure in human plasma.

[0169] 2. Experimental Method:

[0170] 5 μL of samples with concentrations of 20, 50, 100, 200, 500, 1000, 2000, 5000, and 10000 ng / mL were added to 45 μL of human plasma, and the content of the compounds was detected by LC-MS to form a standard curve. 5 μL of a polypeptide solution with a concentration of 1 mg / mL was added to 45 μL of human plasma. Five samples were prepared for each compound to be tested, and one sample was taken out at 0 minutes, 30 minutes, 60 minutes, 120 minutes, and 240 minutes, respectively, to detect the content of the remaining compounds by LC-MS, and the content of the remaining compounds at other time points was calculated based on the content of the remaining compounds at 0 minutes (100%). The method for detecting the content of the compounds by LC-MS was as follows: 5% acetonitrile solution was prepared as solution A, 95% acetonitrile solution was prepared as solution B, the flow rate was 0.6 mL / min, the time and solution ratio shown in Table 9 were used to form a solution gradient, 15 μL of sample was injected, and the content of the compounds was detected by Raptor Biphenyl 2.7 micron detection column, as shown in Table 9.

[0171] Table 9. Detection time points and solution ratios

[0172] Time (min) A (%) B (%) 0.2 95.0 5.0 1.7 95.0 9.5 0.2 95.0 2.0 5.0 95.0 5.0 2.5 95.0 5.0 0.0

[0173] 3. Experimental results:

[0174] The stability data of some of the compounds of the present disclosure in plasma is shown in Table 10 below.

[0175] Table 10. Experimental results of the stability of the compounds in plasma

[0176]

[0177]

[0178] Experimental conclusion:

[0179] It was found through research that the compound 7# of the present disclosure has similar stability (relative content > 90%) in human plasma at a 4-hour time point compared to the compound LY3298176.

[0180] Example 6 Pharmacokinetic properties of some of the compounds of the present disclosure in mice

[0181] Plasma stability is one of the factors that affect the pharmacokinetics of polypeptide drugs. The pharmacokinetics of polypeptide drugs in vivo is also affected by factors such as their absorption and clearance in vivo.

[0182] 1. Experimental purpose:

[0183] The purpose of this experiment is to study the pharmacokinetic behavior of the compound of the present disclosure in mice (plasma) after a single intravenous injection of the drug using Balb / c mice as test animals.

[0184] 2. Experimental method:

[0185] Male Balb / c mice aged 7-9 weeks with body weight of 18-30 g were purchased from Shanghai Jiesijie Laboratory Animal Co., Ltd. After compound 7# was prepared in 20 mM citric acid buffer (pH = 7.0), compound 7# was injected into mice through the tail vein at a dose of 30 nmol / kg of body weight, and 0.2 mL of blood was collected at time points of 0 h, 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h and 32 h. The collected mouse blood was centrifuged at a speed of 6000 rpm for 6 min at a temperature of 4°C to separate the plasma. The content of compound 7# in the mouse plasma was detected by the experimental method of Example 3.3.

[0186] 3. Experimental results:

[0187] Through the above experimental method, the specific data are as shown in Table 11:

[0188] Table 11. Pharmacokinetic behavior of single intravenous injection administration in mice (plasma)

[0189] PK parameter unit Compound 7# T 1 / 2 h 13.0 AUC Inf h*ng / mL 16133

[0190] 4. Experimental conclusion:

[0191] It is found through research that compound 7# of the present disclosure has good pharmacokinetic characteristics after intravenous injection administration in mice, indicating that it has advantages in treating diseases, for example, it can support once-a-week subcutaneous injection administration in humans.

[0192] Example 7. Pharmacokinetic characteristics of some compounds of the present disclosure in mice

[0193] 1. Experimental purpose:

[0194] The purpose of this experiment is to study the pharmacokinetic behavior of single subcutaneous injection administration of the compounds of the present disclosure in mice (plasma) by taking Balb / c mice as the test animals.

[0195] 2. Experimental method:

[0196] Male Balb / c mice aged 7-9 weeks with body weight of 18-30 g were purchased from Shanghai Jiesijie Experimental Animal Co., Ltd. After compound 7# was prepared in 20 mM citric acid buffer (pH = 7.0), compound 7# was injected into mice through left abdominal subcutaneous injection at a dose of 30 nmol / kg of body weight, and 0.2 mL of blood was collected at time points of 0 h, 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h and 32 h. The collected mouse blood was centrifuged at a speed of 6000 rpm for 6 min at a temperature of 4 ℃ to separate plasma. The content of compound 7# in mouse plasma was detected by the experimental method of Example 5.2.

[0197] 3. Experimental results:

[0198] Through the above experimental method, the specific data are shown in Table 12:

[0199] Table 12. Pharmacokinetic results of compound 7# in mice

[0200] PK parameter unit Compound 7# T 1 / 2 h10.1 AUC Inf h*ng / mL 14488

[0201] 4. Experimental conclusion:

[0202] It is found through research that the compound of the present disclosure has good pharmacokinetic characteristics after subcutaneous injection administration in mice, indicating that it has advantages in treating diseases, for example, it can support once-a-week subcutaneous injection administration in humans.

[0203] Example 8 In vivo efficacy of part of the compounds of the present disclosure

[0204] 1. Experimental purpose:

[0205] The regulation effect of single subcutaneous administration of part of the compounds of the present disclosure and compound LY3298176 on blood glucose of normal mice was tested.

[0206] 2. Experimental method:

[0207] Male C57BL / 6 mice aged 10-12 weeks were purchased from Shanghai Jiesijie Experimental Animal Co., Ltd. After C57BL / 6 mice were subcutaneously injected with compound 7# or compound LY3298176 (dose: 10 nmol / kg body weight) and control buffer, they were fasted and not water-restricted, and 18 hours later, a glucose solution with a concentration of 0.2 g / mL was injected intraperitoneally. According to the experimental design, blood glucose values were measured at 0 minutes, 15 minutes, 30 minutes, 60 minutes, and 120 minutes by taking blood from the tail of the mouse. The specific method is to fix the mouse with a physical method, expose the tail and cut off a little of the tail, squeeze the tail to make it bleed, discard the first drop of blood, and then use a Roche active blood glucose meter to detect blood glucose. According to the results at each point, the area under the blood glucose curve (AUC) was calculated.

[0208] 3. Experimental results:

[0209] Through the above experimental method, the specific data are shown in Table 13 below:

[0210] Table 13. Changes in blood glucose values of mice after single subcutaneous administration

[0211]

[0212] 4. Experimental conclusion:

[0213] In this experiment, at a dose of 10 nmol / kg body weight, the compound 7# of the present disclosure showed a significant hypoglycemic effect on normal mice, and the area under the blood glucose curve of the compound 7# group decreased by more than 60% compared with the placebo (i.e., blank solvent).

[0214] Example 9 Weight loss efficacy of some compounds of the present disclosure

[0215] 1. Purpose of the experiment:

[0216] Test number Compound Subcutaneous administration of diet-induced obese mice to regulate the effect of body weight.

[0217] 2. Experimental method:

[0218] High-fat diet induced obese male C57BL / 6 mice (body weight 35-55 g, 10-12 weeks old, purchased from Shanghai Jiesijie Experimental Animal Co., Ltd.). The diet-induced obese C57BL / 6 mice were subcutaneously injected with compound LY3298176 (10 nmol / kg body weight), 7# (10 nmol / kg body weight), 18# (three doses, 3, 10, and 100 nmol / kg body weight, respectively, administered every 3 days). According to the experimental design, the body weight of each mouse was weighed and recorded on day 0, day 3, day 6, and so on to day 27, the average body weight of each group of mice was calculated, and the body weight change curve was plotted based on the first day body weight. After the end, the fat and other organs of the mice were weighed, the fat organ / brain ratio of each mouse was calculated, and the effect of the drug on fat was determined by comparing the changes in the fat organ / brain ratio of each group of mice in different parts.

[0219] 3. Experimental results:

[0220] Through the above experimental method, the specific data are shown in Tables 14 to 16 and FIG. 1.

[0221] Table 14. Effect of compounds on weight loss of obese mice induced by high-fat diet

[0222]

[0223]

[0224]

[0225]

[0226] 4. Experimental conclusion:

[0227] In this experiment, the compounds 7# and 18# of the present disclosure showed a significant weight loss effect on high-fat diet-induced obese mice at doses of 3 nmol / kg, 10 nmol / kg, and 100 nmol / kg, and showed a significant dose-dependent effect. The body weight of the test group of compound 18# at a dose of 10 nmol / kg decreased by more than 20.0% on day 27, while the body weight of the test group of the control compound LY3298176 under the same dose condition decreased by about 13.4%. In addition, the fat content of each part of the test group of compound 18# at different doses was significantly reduced compared with the placebo (i.e., blank solvent) group.

[0228] Example 10. Effect of the compound of the present disclosure on food intake of mice

[0229] The food intake of each group of mice was measured daily during the experiment, and the results are shown in Table 17 and FIG. 2.

[0230] The average daily food intake of the model control DIO (diet-induced obsisity) mice was 2.5 g during the whole experiment. After subcutaneous injection of different doses of compound 18# or LY3298176, the food intake of mice in each group decreased to different degrees.

[0231] On the first day after administration, the food intake of mice in each administration group decreased significantly. The food intake of mice in the 3, 10 and 100 nmol / kg compound 18# groups was 0.6 g, 0.3 g and 0.2 g, respectively, which was significantly different from that of the model control group (2.5 g), and showed a good dose-effect relationship.

[0232] The cumulative food intake of mice in the model control group within 5 days after administration was 12.8 g, while the cumulative food intake of mice in the 3, 10 and 100 nmol / kg compound 18# groups within 5 days after administration was 7.2 g, 3.9 g and 1.8 g, respectively, which was significantly lower than that of the model control group, and showed a good dose-effect relationship.

[0233] The daily food intake of each administration group began to decrease on the first day after each administration, and began to recover on the second and third days. The daily food intake showed an overall upward recovery trend during the administration period. After 28 days of administration, the cumulative food intake of the three dose groups of compound 18# was 58.2 g, 46.8 g and 36.7 g, respectively, which was significantly lower than that of the model control group (70.8 g), and showed a good dose-dependent relationship. Therefore, compound 18# can significantly reduce the food intake of DIO mice.

[0234]

[0235]

[0236] Example 11 Improvement effect of the compound of the present disclosure on the glucose metabolism level of db / db mice

[0237] 1. Purpose of the experiment:

[0238] Test the improvement effect of subcutaneous administration of the compound on the glucose metabolism level of db / db mice.

[0239] 2. Experimental method:

[0240] C57BL / KsJ-db / db mice were subcutaneously injected with blank vehicle (20 mM sodium citrate + 0.05% Tween-80, pH 7.5), compound LY3298176 (100 nmol / kg body weight), and 18# (three doses, 10, 30, 100 nmol / kg body weight, respectively) on days 0, 3, 7, 10, 14, 17, 21, 24, and 27. Each administration group contained 10 db / db mice. According to the experimental design, on days 0, 7, 14, 21, and 28, tail vein blood was pricked for fasting blood glucose level detection using a blood glucose meter and blood glucose test paper, and the animals were fasted for 6 hours before each blood collection point. On days 3, 10, 17, 24, and 27, tail vein blood was pricked for random blood glucose level detection using a blood glucose meter. Finally, after the experiment ended on day 28, all animals in each administration group were anesthetized by inhaling 2-5% isoflurane, and 100 μl of EDTA-K2 anticoagulant whole blood was collected through the orbit for determination of glycated hemoglobin.

[0241] 3. Experimental results:

[0242] Through the above experimental method, the specific data are shown in Tables 18-20 below

[0243] Table 18. Effect of long-term administration of compound 18# on fasting blood glucose in db / db mice

[0244]

[0245] ***: p < 0.001 vs. blank control group.

[0246] Table 19. Effect of long-term administration of compound 18# on random blood glucose in db / db mice

[0247]

[0248]

[0249] **: p < 0.01 vs. blank control group; ***: p < 0.001 vs. blank control group.

[0250] Table 20. Effect of long-term administration of 18# on glycated hemoglobin level in db / db mice

[0251] Dose group Glycated hemoglobin (%) (mean ± SD) Blank control 6.54 ± 0.17 LY3298176 (100 nmol / kg) 4.58 ± 0.23**#18 (10 nmol / kg) 4.71 ± 0.23***#18 (30 nmol / kg) 4.53 ± 0.17***#18 (100 nmol / kg) 3.78 ± 0.13***

[0252] **: p < 0.01 vs blank control; ***: p < 0.001 vs blank control.

[0253] 4. Experimental Conclusion

[0254] In the present experiment, the compound 18# of the present disclosure showed an excellent improvement effect on the level of glucose metabolism of db / db mice at the doses of 10 nmol / kg, 30 nmol / kg and 100 nmol / kg, and presented a significant dose correlation. The level of glycated hemoglobin of the test group of compound 18# at the dose of 100 nmol / kg was 3.78% at the end of the experiment, while the level of glycated hemoglobin of the test group of the control compound LY3298176 under the same dose condition was 4.58%. The efficacy of compound 18# in improving the level of glucose metabolism of db / db mice was significantly superior to that of the control compound LY3298176 under the same dose condition.

Claims

1. A GLP-1 analogue or a pharmaceutically acceptable salt thereof, wherein the GLP-1 analogue has the general formula (I): R 1 -X 1 -X 2 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -Glu-Phe-X 23 -X 24 -Trp-Leu-X 27 -X 28 -X 29 -X 30 -Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R 2 (I) in ： R 1 is H, alkyl, acetyl, formyl, benzoyl, trifluoroacetyl, pGlu or absent; R 2 Yes-NH 2 , -OH or not present; X 1 , X 2 , X 10 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 23 , X 24 , X 27 , X 28 , X 29 and X 30 Independently selected from any natural amino acid residue or non-natural amino acid residue.

2. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr or His; X 2 An amino acid residue selected from Aib or D-Ala; X 10 An amino acid residue selected from Val or Tyr or Y1; X 12 An amino acid residue selected from Ser or Ile or Y1; X 13 An amino acid residue selected from Tyr or Ala or Y1; X 14 An amino acid residue selected from Leu or Nle or Y1; X 15 An amino acid residue selected from Asp or Glu; X 16 An amino acid residue selected from Arg, Glu, Gly, Lys or Aib or Y1; X 17 An amino acid residue selected from Glu, Ile or Gln or Y1; X 18 An amino acid residue selected from Ala, Aib or His; X 19 An amino acid residue selected from Ala, Aib or Gln; X 20 An amino acid residue selected from Gln, Glu, Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Ala, Asn or Gln; X 27 An amino acid residue selected from Val or Leu; X 28 An amino acid residue selected from Arg or Ala; X 29 An amino acid residue selected from Gly or Gln; X 30 An amino acid residue selected from Gly and Lys; Y1 is a Lys, Orn, Dap, Dab or Cys residue comprising a substituent on the side chain having the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH; a is an integer from 1 to 3; b is 1 or 2; c is an integer from 10 to 30.

3. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib; X 10 An amino acid residue selected from Tyr; X 12 An amino acid residue selected from Ile; X 13 An amino acid residue selected from Tyr; X 14 Select from Y1;X 15 An amino acid residue selected from Asp or Glu; X 16 An amino acid residue selected from Arg or Lys; X 17 An amino acid residue selected from Ile; X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

4. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 3, It is characterized in that X 16 An amino acid residue selected from Lys; X 23 An amino acid residue selected from Val; X 27 An amino acid residue selected from Leu.

5. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib or D-Ala; X 10 Select from Y1;X 12 An amino acid residue selected from Ile; X 13 An amino acid residue selected from Tyr; X 14 An amino acid residue selected from Leu or Nle; X 15 An amino acid residue selected from Glu; X 16 An amino acid residue selected from Arg or Lys; X 17 An amino acid residue selected from Ile; X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln or Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn or Gln; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

6. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib or D-Ala; X 10 An amino acid residue selected from Tyr; X 12 Select from Y1;X 13 An amino acid residue selected from Tyr; X 14 An amino acid residue selected from Leu or Nle; X 15 An amino acid residue selected from Glu; X 16 An amino acid residue selected from Arg or Lys; X 17 An amino acid residue selected from Ile; X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln or Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn or Gln; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

7. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib or D-Ala; X 10 An amino acid residue selected from Tyr; X 12 An amino acid residue selected from Ile; X 13 Select from Y1;X 14 An amino acid residue selected from Leu or Nle; X 15 An amino acid residue selected from Glu; X 16 An amino acid residue selected from Arg or Lys; X 17 An amino acid residue selected from Ile; X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln or Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn or Gln; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

8. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib or D-Ala; X 10 An amino acid residue selected from Tyr; X 12 An amino acid residue selected from Ile; X 13 An amino acid residue selected from Tyr; X 14 Select from Y1;X 15 An amino acid residue selected from Glu; X 16 An amino acid residue selected from Arg or Lys; X 17 An amino acid residue selected from Ile; X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln or Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn or Gln; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

9. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 8, It is characterized in that X 2 An amino acid residue selected from Aib; X 20 An amino acid residue selected from Gln; X 24 An amino acid residue selected from Asn.

10. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib or D-Ala; X 10 An amino acid residue selected from Tyr; X 12 An amino acid residue selected from Ile; X 13 An amino acid residue selected from Tyr; X 14 An amino acid residue selected from Leu or Nle; X 15 An amino acid residue selected from Glu; X 16 Select from Y1;X 17 An amino acid residue selected from Ile; X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln or Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn or Gln; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

11. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 10, It is characterized in that X 2 An amino acid residue selected from Aib; X 14 An amino acid residue selected from Leu; X 20 An amino acid residue selected from Gln; X 24 An amino acid residue selected from Asn.

12. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that X 1 An amino acid residue selected from Tyr; X 2 An amino acid residue selected from Aib or D-Ala; X 10 An amino acid residue selected from Tyr; X 12 An amino acid residue selected from Ile; X 13 An amino acid residue selected from Tyr; X 14 An amino acid residue selected from Leu or Nle; X 15 An amino acid residue selected from Glu; X 16 An amino acid residue selected from Arg or Lys; X 17 Select from Y1;X 18 An amino acid residue selected from Ala; X 19 An amino acid residue selected from Ala; X 20 An amino acid residue selected from Gln or Lys; X 23 An amino acid residue selected from Ile or Val; X 24 An amino acid residue selected from Asn or Gln; X 27 An amino acid residue selected from Ile or Leu; X 28 An amino acid residue selected from Ala; X 29 An amino acid residue selected from Gly; X 30 an amino acid residue selected from Gly; Y1 is as defined in claim 2.

13. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 2 to 12, It is characterized in that a is 2, b is 1 or 2, and c is an integer from 16 to 20.

14. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 13, It is characterized in that c is 16, 18 or 20.

15. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 2 to 14, It is characterized in that Y1 is a Lys residue comprising a substituent on the side chain having the formula {[2-(2-amino-ethoxy)-ethoxy]-acetyl} a -(y-Glu) b -CO-(CH 2 ) c -COOH; a is 2; b is 1 or 2; c is 16 or 18.

16. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 2 to 15, It is characterized in that The substituent is covalently linked to the amino group on the side chain via an amide bond.

17. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 2 to 16, It is characterized in that Y1 is K(-OEG-OEG-yGlu-C18-OH) or K(-OEG-OEG-yGlu-C20-OH), wherein K(-OEG-OEG-yGlu-C18-OH) has the following structure: Preferably, it has the following structure: K(-OEG-OEG-yGlu-C20-OH) has the following structure: Preferably, it has the following structure:

18. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 2 to 17, It is characterized in that The substituent is covalently linked to the epsilon amino group on the side chain via an amide bond.

19. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that The GLP-1 analog is shown in SEQ ID NO: 20; Preferably, the GLP-1 analogue is selected from the following compounds No. 1-18:

20. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that The GLP-1 analogue is selected from the following compounds numbered 1#-18#:

21. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that The GLP-1 analog is selected from the compounds shown in 7#, 12#, 13#, 14#, 15#, 16#, 17#, or 18# in FIG. 3 .

22. The GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 21, wherein the agonist activity of the GLP-1 analogue or a pharmaceutically acceptable salt thereof on the GLP-1R is higher than the agonist activity on the GIP receptor; Preferably, the ratio of the agonist activity of the GLP-1 analogue on GLP-1R to the agonist activity of the GIP receptor is 1.1:1 to 10:1, more preferably 3:1 to 6.5:1, and most preferably 4.5:1 to 6:

1.

23. A pharmaceutical composition comprising: 1) A GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 22, and 2) Pharmaceutically acceptable excipients or drug carriers.

24. Use of the GLP-1 analogue or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 22, and the composition according to claim 23, in the preparation of a medicament for treating non-insulin-dependent diabetes mellitus, insulin-dependent diabetes mellitus, obesity, non-alcoholic fatty liver disease, hepatic steatosis, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, insulin resistance, dyslipidemia associated with insulin resistance, and / or dyslipidemia associated with diabetes mellitus.