Polypeptide agonist, preparation method therefor and pharmaceutical use thereof

Abstract

The present invention relates to a polypeptide agonist, a preparation method therefor and the pharmaceutical use thereof. Specifically, the present invention relates to a polypeptide as represented by general formula (I) and a derivative or pharmaceutically acceptable salt thereof, a pharmaceutical composition thereof, and the use thereof in the preparation of a drug for preventing or treating a disease or disorder related to metabolic disorders.

Description

Polypeptide agonists, methods of making and medical uses thereof TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to a kind of polypeptide and its derivative, or its pharmaceutically acceptable salt, which can be used as an agonist for preventing or treating metabolic disorder related diseases or conditions. BACKGROUND

[0002] Diabetes is a metabolic disease caused by insufficient insulin secretion in the body, which further leads to the disorder of glucose, protein and lipid metabolism in 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 the global diabetic patients are non-insulin-dependent diabetes. Non-insulin-dependent diabetes is a long-term and 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 plasma. Studies have shown that non-insulin-dependent diabetes is related to a variety of high-risk diseases in patients, and it often leads to patients suffering from cardiovascular diseases, kidney failure, blindness, amputation and other various diseases.

[0003] 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 diseases, 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.

[0004] 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 such as recombinant insulin and its derivatives produced by biosynthesis. 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 and development direction in clinical practice.

[0005] 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 the 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, while 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 highly 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, which means 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 and its analogs mean that GLP-1 polypeptide is an ideal drug for treating non-insulin-dependent diabetes and obesity diabetes.

[0006] 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, but human natural GLP-1 has poor drug properties and is easily degraded by dipeptidyl peptidase-IV (DPP-IV) in the body, so 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 enzyme cleavage site amino acid, fatty acid modification of the polypeptide backbone, and combination of GLP-1 polypeptide and various proteins / polymer polymers. The long-acting GLP-1 analogs that have been marketed and widely used in clinical practice at this stage include exenatide for subcutaneous injection twice a day, liraglutide for subcutaneous injection once a day, and dulaglutide and semaglutide for subcutaneous injection once a week, etc.

[0007] 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, GLP-1 polypeptides and their derivatives have been found to cause the heartbeat of the subject to accelerate and, in certain cases, to 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 their clinical use cannot achieve full blood glucose control and weight loss in patients.

[0008] Glucose-dependent insulinotropic polypeptide (GIP) and GLP-1 polypeptide belong to one of the incretins, which play a key physiologically relevant 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 cell near the jejunum according to the glucose level in the plasma. GIP polypeptide exerts its physiological effect by combining with 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 its GIP receptor in adipose tissue to promote fat metabolism. Interestingly, intracerebroventricular injection of GIP polypeptide in mice can reduce food intake and body weight in test animals, which seems to indicate that GIP polypeptide also has certain specific physiological functions in reducing body weight. Studies have shown that the incretin function of GIP polypeptide in non-insulin-dependent diabetes patients 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 weakened when the blood glucose level returns to normal.

[0009] Glucagon (GCG) is a 29-amino acid linear peptide secreted by pancreatic islet alpha cells. Glucagon receptor (GCGR) is expressed in the liver and kidney, and also in the heart, adipocytes, spleen, pancreas, brain, gastrointestinal tract, etc. GCG and insulin work together to maintain the homeostasis of blood glucose in the body. Hypoglycemia stimulates the pancreatic alpha cells to secrete GCG, which activates the glucagon receptor in the liver, activates the downstream cAMP / PKA signaling pathway, promotes hepatic gluconeogenesis and glycogenolysis, reduces the synthesis of liver glycogen, increases the output of liver glucose, and increases the blood glucose level. Injection of GCG in humans and animals has been shown to increase blood glucose levels. In addition, GCG can improve energy metabolism and heat production in the body, and GCG can increase lipid decomposition in white adipose tissue and improve systemic lipid metabolism. In particular, GCG can enhance the level of lipid metabolism in the liver, and activation of liver GCGR can lead to activation of the lipid metabolism pathway in hepatocytes. GCG has also been shown to have an inhibitory effect on food intake. GLP-1 / GCG plays different and complementary roles in different metabolic organs and tissues. In addition to lowering blood sugar and weight loss, GLP-1 / GCG can increase brain satiety, inhibit food intake, increase leptin sensitivity, increase energy consumption in adipose tissue, stimulate lipolysis in adipose tissue, fatty acid oxidation, etc.

[0010] Currently, LY3437943 polypeptide invented by Lilly is a GLP-1R / GIPR / GCGR triple-target agonist, which is in clinical phase III. Once-weekly subcutaneous injection shows significant efficacy in the treatment of type II diabetes and weight loss, and the weight loss effect is generally better than that of single-target and double-target therapies based on GLP1R. It also has great development potential in indications such as non-alcoholic fatty liver, and its safety and tolerability in the body are similar to other incretins, which has good development prospects. The purpose of the present application is to develop a GLP-1R / GIPR / GCGR triple-target agonist that can significantly achieve weight loss and blood glucose control. SUMMARY

[0011] The purpose of the present application is to provide a new class of polypeptides and derivatives thereof, or pharmaceutically acceptable salts thereof, which can be used as GLP-1 receptor, GIP receptor and / or GCG receptor agonists.

[0012] Specifically, the present application comprises a polypeptide and derivatives thereof as shown in general formula (I0), or a pharmaceutically acceptable salt thereof, R1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16-X 17 -X 18 -X 19 -X 20 -X 21 -X 22 -X 23 -X 24 -X 25 -X 26 -X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 -X 36 -X 37 -X 38 -X 39 -R2(I0)

[0013] wherein:

[0014] R1is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acetyl, substituted or unsubstituted formyl, or substituted or unsubstituted benzoyl;

[0015] R2is selected from -NH2or -OH;

[0016] X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 or X 39each independently present or absent, and if present, each independently selected from the group consisting of any modified or unmodified natural amino acid residue, modified or unmodified non-natural amino acid residue, or a peptide segment consisting thereof;

[0017] said modified natural amino acid residue optionally can be attached to a side chain;

[0018] said modified non-natural amino acid residue optionally can be attached to a side chain;

[0019] or, any two adjacent or non-adjacent amino acid residues can optionally be further attached to form a modified or unmodified ring.

[0020] In a further preferred embodiment of the present application, said X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 or X 39 are each independently selected from the group consisting of any modified or unmodified natural or non-natural amino acid residue.

[0021] In a further preferred embodiment of the present application, said X1is selected from the group consisting of a modified or unmodified Tyr or His amino acid residue; preferably from a Tyr or His amino acid residue; more preferably from a His amino acid residue.

[0022] In a further preferred embodiment of the present application, said X2is selected from the group consisting of a modified or unmodified Aib, Ala or Ser amino acid residue; preferably from a Aib, Ala or Ser amino acid residue; more preferably from a Aib amino acid residue.

[0023] In a further preferred embodiment of the present application, X3 is selected from the group consisting of a modified or unmodified amino acid residue of Glu, Gin or His; preferably from a modified or unmodified amino acid residue of Glu, Gin or His; more preferably from a modified or unmodified amino acid residue of His.

[0024] In a further preferred embodiment of the present application, X4 is selected from the group consisting of a modified or unmodified amino acid residue of Gly; preferably from a modified or unmodified amino acid residue of Gly.

[0025] In a further preferred embodiment of the present application, X5 is selected from the group consisting of a modified or unmodified amino acid residue of Thr; preferably from a modified or unmodified amino acid residue of Thr.

[0026] In a further preferred embodiment of the present application, X6 is selected from the group consisting of a modified or unmodified amino acid residue of Phe; preferably from a modified or unmodified amino acid residue of Phe.

[0027] In a further preferred embodiment of the present application, X7 is selected from the group consisting of a modified or unmodified amino acid residue of Ile or Thr; preferably from a modified or unmodified amino acid residue of Thr.

[0028] In a further preferred embodiment of the present application, X8 is selected from the group consisting of a modified or unmodified amino acid residue of Ser; preferably from a modified or unmodified amino acid residue of Ser.

[0029] In a further preferred embodiment of the present application, X9 is selected from the group consisting of a modified or unmodified amino acid residue of Asp; preferably from a modified or unmodified amino acid residue of Asp.

[0030] In a further preferred embodiment of the present application, X 10 is selected from the group consisting of a modified or unmodified amino acid residue of Tyr, Val, Leu, Fae or Lys; preferably from a modified or unmodified amino acid residue of Tyr, Fae or Lys.

[0031] In a further preferred embodiment of the present application, X 11 is selected from the group consisting of a modified or unmodified amino acid residue of Ser; preferably from a modified or unmodified amino acid residue of Ser.

[0032] In a further preferred embodiment of the present application, X 12 is selected from the group consisting of a modified or unmodified amino acid residue of Ile, Lys or Ser; preferably from a modified or unmodified amino acid residue of Ile or Lys.

[0033] In a further preferred embodiment of the present application, X 13an amino acid residue selected from Ala, Tyr, Aib, Leu, Achx, α-Me-cpAla, 4diFAchx, THP, THT, THS or Lys, preferably from Leu or α-Me-Leu, or, preferably from α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT, THS or modified Lys.

[0034] In a further preferred embodiment of the application, said X 14 an amino acid residue selected from Met or Leu, preferably from Met or Leu, more preferably from Leu.

[0035] In a further preferred embodiment of the application, said X 15 an amino acid residue selected from Asp or Glu, preferably from Asp or Glu, more preferably from Glu.

[0036] In a further preferred embodiment of the application, said X 16 an amino acid residue selected from Lys, Ser or Gly, preferably from modified or unmodified Lys.

[0037] In a further preferred embodiment of the application, said X 17 an amino acid residue selected from Ile, Arg, Gin or Lys, preferably from modified or unmodified Lys.

[0038] In a further preferred embodiment of the application, said X 18 an amino acid residue selected from His, Arg or Ala, preferably from Ala.

[0039] In a further preferred embodiment of the application, said X 19 an amino acid residue selected from Gin, Lys or Ala, preferably from Ala or modified Lys.

[0040] In a further preferred embodiment of the application, said X 20 an amino acid residue selected from Gin, Lys or Aib, preferably from Gin or modified Lys.

[0041] In a further preferred embodiment of the application, said X 21an amino acid residue selected from modified or unmodified Asp, Glu, Ala or Lys; preferably an amino acid residue selected from Glu or modified Lys.

[0042] In a further preferred embodiment of the application, said X 22 an amino acid residue selected from modified or unmodified Phe; preferably an amino acid residue selected from Phe.

[0043] In a further preferred embodiment of the application, said X 23 an amino acid residue selected from modified or unmodified Val or lie; preferably an amino acid residue selected from Val.

[0044] In a further preferred embodiment of the application, said X 24 an amino acid residue selected from modified or unmodified Asn, Gin, Ala or Glu; preferably an amino acid residue selected from Glu.

[0045] In a further preferred embodiment of the application, said X 25 an amino acid residue selected from modified or unmodified Trp or Tyr; preferably an amino acid residue selected from Trp.

[0046] In a further preferred embodiment of the application, said X 26 an amino acid residue selected from modified or unmodified Leu; preferably an amino acid residue selected from Leu.

[0047] In a further preferred embodiment of the application, said X 27 an amino acid residue selected from modified or unmodified Leu, Met, Val or lie; preferably an amino acid residue selected from Leu.

[0048] In a further preferred embodiment of the application, said X 28 an amino acid residue selected from modified or unmodified Ala, Asn, Lys, Arg or Glu; preferably an amino acid residue selected from Ala.

[0049] In a further preferred embodiment of the application, said X 29 an amino acid residue selected from modified or unmodified Gin, Thr or Gly; preferably an amino acid residue selected from Gly.

[0050] In a further preferred embodiment of the application, said X 30 an amino acid residue selected from modified or unmodified Lys, Arg, Gly or is absent; preferably an amino acid residue selected from Gly.

[0051] In a further preferred embodiment of the application, said X31 an amino acid residue selected from modified or unmodified Pro, Gly, or is absent; preferably an amino acid residue selected from Pro.

[0052] In a further preferred embodiment of the application, said X 32 an amino acid residue selected from modified or unmodified Lys, Ser, or is absent; preferably an amino acid residue selected from Ser.

[0053] In a further preferred embodiment of the application, said X 33 an amino acid residue selected from modified or unmodified Lys, Ser, or is absent; preferably an amino acid residue selected from Ser.

[0054] In a further preferred embodiment of the application, said X 34 an amino acid residue selected from modified or unmodified Asn, Gly, or is absent; preferably an amino acid residue selected from Gly.

[0055] In a further preferred embodiment of the application, said X 35 an amino acid residue selected from modified or unmodified Asp, Ala, or is absent; preferably an amino acid residue selected from Ala.

[0056] In a further preferred embodiment of the application, said X 36 an amino acid residue selected from modified or unmodified Trp, Pro, or is absent; preferably an amino acid residue selected from Pro.

[0057] In a further preferred embodiment of the application, said X 37 an amino acid residue selected from modified or unmodified Lys, Pro, or is absent; preferably an amino acid residue selected from Pro.

[0058] In a further preferred embodiment of the application, said X 38 an amino acid residue selected from modified or unmodified His, Pro, or is absent; preferably an amino acid residue selected from Pro.

[0059] In a further preferred embodiment of the application, said X 39 an amino acid residue selected from modified or unmodified Asn, Ser, or is absent; preferably an amino acid residue selected from Ser.

[0060] The present application further provides a polypeptide according to general formula (I-0) and derivatives thereof, or a pharmaceutically acceptable salt thereof, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-X9-X 10 -Ser-X 12 -X 13-Leu-X 15 -X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-X 23 -Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2 (I-0)

[0061] wherein:

[0062] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acetyl, substituted or unsubstituted formyl or substituted or unsubstituted benzoyl;

[0063] R2 is selected from -NH2 or -OH;

[0064] X9 is selected from Asp, Glu, Asn, Gin, Lys, Arg, THP, Ser, Thr;

[0065] X 10 an amino acid residue selected from Tyr, Fae, modified or unmodified Lys;

[0066] X 12 an amino acid residue selected from lie or modified or unmodified Lys;

[0067] X 13 an amino acid residue selected from Leu, α-Me-Leu, α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT, THS or modified or unmodified Lys;

[0068] X 15 selected from Glu, Asp, Asn, Gin, Lys, Arg, THP, Ser, Thr;

[0069] X 16 an amino acid residue selected from modified or unmodified Lys;

[0070] X 17 an amino acid residue selected from modified or unmodified Lys;

[0071] X 19 an amino acid residue selected from Ala or modified or unmodified Lys;

[0072] X 20 an amino acid residue selected from Gin or modified or unmodified Lys;

[0073] X 21 an amino acid residue selected from the group consisting of Glu or modified or unmodified Lys;

[0074] X 23 an amino acid residue selected from the group consisting of Ile, Val, Leu, Ala, Aib, Achx, α-Me-Leu, α-Me-cpAla;

[0075] or, any two adjacent or non-adjacent amino acid residues can optionally be further linked to form a modified or unmodified ring.

[0076] The present application further provides a polypeptide as shown in general formula (I-1) and its derivatives, or a pharmaceutically acceptable salt thereof, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-X9-X 10 -Ser-X 12 -X 13 -Leu-Glu-X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-1).

[0077] The present application further provides a polypeptide as shown in general formula (I-2) and its derivatives, or a pharmaceutically acceptable salt thereof, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -Leu-X 15 -X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-2).

[0078] The present application further provides a polypeptide as shown in general formula (I-3) and its derivatives, or a pharmaceutically acceptable salt thereof, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -Leu-Glu-X16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-X 23 -Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-3).

[0079] The present application further provides a polypeptide as shown in general formula (I-4) and its derivatives, or its pharmaceutically acceptable salt, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Ser-X9-X 10 -Ser-X 12 -X 13 -Leu-X 15 -X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-X 23 -Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-4).

[0080] The present application further provides a polypeptide as shown in general formula (I-5) and its derivatives, or its pharmaceutically acceptable salt, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-X9-X 10 -Ser-X 12 -X 13 -Leu-X 15 -X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-5).

[0081] The present application further provides a polypeptide as shown in general formula (I-6) and its derivatives, or its pharmaceutically acceptable salt, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-X9-X 10 -Ser-X 12 -X 13 -Leu-Glu-X 16 -X17 -Ala-X 19 -X 20 -X 21 -Phe-X 23 -Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-6).

[0082] The present application further provides a polypeptide according to Formula (I-A) and derivatives thereof, or a pharmaceutically acceptable salt thereof, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -Leu-Glu-X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(I-A)

[0083] wherein:

[0084] R1 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acetyl, substituted or unsubstituted formyl, or substituted or unsubstituted benzoyl;

[0085] R2 is selected from -NH2 or -OH;

[0086] X 10 is an amino acid residue selected from Tyr, Fae, modified or unmodified Lys;

[0087] X 12 is an amino acid residue selected from Ile or modified or unmodified Lys;

[0088] X 13 is an amino acid residue selected from Leu, α-Me-Leu, α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT, THS or modified or unmodified Lys;

[0089] X 16 is an amino acid residue selected from modified or unmodified Lys;

[0090] X 17 is an amino acid residue selected from modified or unmodified Lys;

[0091] X 19 an amino acid residue selected from the group consisting of Ala or modified or unmodified Lys;

[0092] X 20 an amino acid residue selected from the group consisting of Gln or modified or unmodified Lys;

[0093] X 21 an amino acid residue selected from the group consisting of Glu or modified or unmodified Lys;

[0094] or, any two adjacent or non-adjacent amino acid residues can optionally be further linked to form a modified or unmodified ring.

[0095] The present application further provides a preferred embodiment, a polypeptide according to the general formula (I-A) and derivatives thereof, or pharmaceutically acceptable salts thereof, wherein each amino acid position of the general formula (I-A) is optionally modified with a side chain; preferably Y2.

[0096] The present application further provides a polypeptide according to the general formula (I) and derivatives thereof, or pharmaceutically acceptable salts thereof, wherein R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -Leu-Glu-X 16 -X 17 -Ala-X 19 -X 20 -X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2 (I)

[0097] wherein:

[0098] R1 is selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acetyl, substituted or unsubstituted formyl or substituted or unsubstituted benzoyl;

[0099] R2 is selected from the group consisting of -NH2 or -OH;

[0100] X 10 an amino acid residue selected from the group consisting of Tyr, Fae, modified or unmodified Lys or Y1;

[0101] X 12 an amino acid residue selected from the group consisting of Ile or modified or unmodified Lys;

[0102] X 13an amino acid residue selected from Leu, α-Me-Leu, α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT, THS or modified or unmodified Lys;

[0103] X 16 an amino acid residue selected from modified or unmodified Lys or Y1;

[0104] X 17 an amino acid residue selected from modified or unmodified Lys;

[0105] X 19 an amino acid residue selected from Ala or modified or unmodified Lys;

[0106] X 20 an amino acid residue selected from Gin or modified or unmodified Lys;

[0107] X 21 an amino acid residue selected from Glu or modified or unmodified Lys;

[0108] Y1is a Fae or Lys residue to which a side chain substituent is attached;

[0109] or, any two adjacent or non-adjacent amino acid residues can optionally be further linked to form a modified or unmodified ring.

[0110] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (II), R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(II)

[0111] wherein:

[0112] X 13 an amino acid residue selected from α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT or THS;

[0113] X 16 is Y1.

[0114] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (III): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-Ile-X 13 -Leu-Glu-Lys-Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(III)

[0115] wherein:

[0116] X 10 is Y1;

[0117] X 13 is selected from the group consisting of Leu, α-Me-Leu, α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT or THS.

[0118] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (IV): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X 12 -X 13 -Leu-Glu-X 16 -Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(IV).

[0119] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (V): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(V).

[0120] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (VII): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-Lys-X 17 -Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(VI).

[0121] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (VII): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-X 19 -Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(VII).

[0122] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (VIII): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-Ala-X 20 -Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(VIII).

[0123] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (IX): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-Lys-X 17 -Ala-Ala-X 20 -Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R(IX).

[0124] In a further preferred embodiment of the application, the polypeptide is further represented by the general formula (X): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-Lys-X 17 -Ala-Ala-Gln-X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2 (X).

[0125] In a further preferred embodiment of the application, the two adjacent or non-adjacent modified amino acid residues are selected from the group consisting of X 16 or X 17 and its adjacent or non-adjacent amino acid residues.

[0126] In a further preferred embodiment of the application, the two adjacent or non-adjacent modified amino acid residues are selected from the group consisting of X 12 and X 16 , X 13 and X 16 , X 13 and X 17 , X 16 and X 19 , X 16 and X 20 , X 17 and X 20 or X 17 and X 21 .

[0127] In a further preferred embodiment of the application, the two adjacent or non-adjacent modified amino acid residues are selected from the group consisting of X

[0128] In a further preferred embodiment of the application, the polypeptide and derivatives thereof comprise a polypeptide sequence represented by any one of the following:

[0129] In a further preferred embodiment of the application, Y1is a Fae or Lys residue to which Y2is attached; Y2is selected from the group consisting of (NEG) a -(OEG) b -(γ-Glu) c -C16diacid, (NEG) a -(OEG) b-(y-Glu) c - C18 diacid, (NEG) a -(OEG) b -(y-Glu) c - C20 diacid, PEG n -(y-Glu) c - C16, (NEG) a -(OEG) b -(y-Glu) c - C24 diacid or (NEG) a -(OEG) b -(y-Glu) c -CO(CH2) 18 PO(OH)2, (NEG) a -(OEG) b -(y-Glu PA) c - C16 diacid, (NEG) a -(OEG) b -(y-Glu PA) c - C18 diacid, (NEG) a -(OEG) b -(y-Glu PA) c - C20 diacid, PEG n -(y-Glu PA) c - C16, (NEG) a -(OEG) b -(y-Glu PA) c - C24 diacid or (NEG) a -(OEG) b -(y-Glu PA) c -CO(CH2) 18 PO(OH)2;

[0130] a is 0, 1, 2, or 3;

[0131] b is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0132] c is 0, 1, 2, or 3;

[0133] n is 16, 18, 20, 22, or 24.

[0134] In further preferred embodiments of the application, any two adjacent or non-adjacent amino acid residues can form a cyclic structure through a modified or unmodified .

[0135] In a further preferred embodiment of the invention, any two adjacent or non-adjacent amino acid residues can be modified or left unmodified. It forms a ring structure.

[0136] In a further preferred embodiment of the present invention, R1 is selected from hydrogen; R2 is selected from -NH2 or -OH.

[0137] In a further preferred embodiment of the present invention, R1 is selected from hydrogen; R2 is selected from -NH2.

[0138] In a further preferred embodiment of the present invention, a is 0 or 1, b is 1, 2 or 3, c is 1, and n is 24.

[0139] In a further preferred embodiment of the present invention, Y1 is covalently linked to Y2 by forming an amide bond from the side chain amino group of Lys; or Y1 is covalently linked to Y2 by forming an amide bond from the side chain amino group of Fae.

[0140] In a further preferred embodiment of the present invention, Y1 is selected from Lys(-NEG-OEG-OEG-γGlu-C16diacid), Lys(-NEG-OEG-OEG-γGlu-C20diacid), Lys(-NEG-OEG-γGlu-C20diacid), Lys(-NEG-OEG-OEG-γGlu-C18diacid), Lys(-OEG-OEG-γGlu-C18diacid), Lys(-OEG-OEG-γGlu-C20diacid), Lys(-PEG). 24 -γGlu-C16), Lys(-NEG-OEG-OEG-OEG-γGlu-C20diacid), Lys(-NEG-OEG-OEG-γGlu-C24diacid), Lys(-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-OEG-γGluPA-C20diacid), Lys(-NEG-OEG-OEG-γGluPA-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-γGlu-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-γGluPA-C20diacid), Lys(-NEG-OEG-γGluPA-CO(CH2) 18PO(OH)2), Lys(-NEG-OEG-OEG-γGlu-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-OEG-γGluPA-C18diacid), Lys(-NEG-OEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-OEG-γGlu-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-OEG-γGluPA-C18diacid), Fae(-NEG-OEG-OEG-γGlu-C16diacid), Fae(-NEG-OEG-OEG-γGlu-C20diacid), Fae(-NEG-OEG-γGlu-C20diacid), Fae(-NEG-OEG-OEG-γGlu-C18diacid), Fae(-OEG-OEG-γGlu-C18diacid), Fae(-OEG-OEG-γGlu-C20diacid), Fae(-PEG 24 -γGlu-C16), Fae(-NEG-OEG-OEG-OEG-γGlu-C20diacid), Fae(-NEG-OEG-OEG-γGlu-C24diacid), Fae(-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2) or Fae(-NEG-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2), the group having the chemical formula of the following structure:

[0141] In a further preferred embodiment of the application, said Y2is selected from the group consisting of -NEG-OEG-OEG-γGlu-C16diacid, -NEG-OEG-OEG-γGlu-C20diacid, -NEG-OEG-γGlu-C20diacid, -NEG-OEG-OEG-γGlu-C18diacid, -OEG-OEG-γGlu-C18diacid, -OEG-OEG-γGlu-C20diacid, -PEG 24- NEG-OEG-OEG-OEG-γGlu-C20 diacid, - NEG-OEG-OEG-γGlu-C24 diacid, - OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2, - NEG-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2, - NEG-OEG-OEG-γGluPA-C20 diacid, - NEG-OEG-OEG-γGluPA-CO(CH2) 18 PO(OH)2, - NEG-OEG-γGlu-CO(CH2) 18 PO(OH) 2、 - NEG-OEG-γGluPA-C20 diacid, - NEG-OEG-γGluPA-CO(CH2) 18 PO(OH) 2、 - NEG-OEG-OEG-γGlu-CO(CH2) 16 PO(OH)2, - NEG-OEG-OEG-γGluPA-C18 diacid, - NEG-OEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2, - NEG-OEG-γGlu-CO(CH2) 16 PO(OH)2, - NEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2or - NEG-OEG-γGluPA-C18 diacid have the chemical formula of the following structure:

[0142] In a further preferred embodiment of the application, the polypeptides according to the application and derivatives thereof, or pharmaceutically acceptable salts thereof, activate the GLP-1 receptor, the GIP receptor and / or the GCG receptor.

[0143] In a further preferred embodiment of the application, the polypeptides according to the application and derivatives thereof, or pharmaceutically acceptable salts thereof, activate the GLP-1 receptor and the GIP receptor.

[0144] In a further preferred embodiment of the application, the polypeptides according to the application and derivatives thereof, or pharmaceutically acceptable salts thereof, activate the GLP-1, GIP and GCG triple receptor.

[0145] The present application further relates to a pharmaceutical composition comprising:

[0146] 1) a therapeutic amount of a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, and

[0147] 2) a pharmaceutically acceptable excipient or a pharmaceutical carrier.

[0148] The present application further provides the use of a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition as described above in the manufacture of a medicament for activating GLP-1 receptor, GIP receptor and / or GCG receptor.

[0149] The present application further provides the use of a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition as described above in the manufacture of a medicament for activating GLP-1, GIP and GCG triple receptor.

[0150] The present application further provides the use of a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, and the composition as described above in the manufacture of a medicament for treating a metabolic disorder-related disease, disorder and / or condition.

[0151] In further preferred embodiments of the present application, the metabolic disorder-related disease, disorder and / or condition is selected from diabetes or a diabetes-related disorder, obesity or an obesity-related disorder, non-alcoholic fatty liver.

[0152] The present application further relates to a method of treating a metabolic disorder-related disease, disorder and / or condition using a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0153] The present application also relates to a method of preventing and / or treating a metabolic disorder-related disease, disorder and / or condition, comprising administering to a patient a therapeutically effective amount of a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0154] In certain embodiments of the present application, the pharmaceutical composition is selected from a tablet, a capsule, a liquid preparation or an injection, preferably further comprising a filler, optionally further comprising a disintegrant, or further comprising one or more of a glidant or a lubricant.

[0155] The present application also provides a method of using a polypeptide and derivatives thereof as described above, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for treating a disease condition, including but not limited to a condition associated with GLP-1, GIP and GCG receptors.

[0156] The present application also relates to a method of treating a metabolic disorder-related condition in a mammal, comprising administering to the mammal a therapeutically effective amount of a polypeptide and derivatives thereof of the present application or a pharmaceutically acceptable salt thereof.

[0157] In some embodiments, the present methods involve treatment of a condition such as a metabolic disorder.

[0158] In some embodiments, the present methods involve diabetes or a diabetes-related condition, obesity or an obesity-related condition, non-alcoholic fatty liver.

[0159] Embodiments of the present application have good physicochemical properties, and exhibit excellent effects in terms of both physical stability and chemical stability, and have good drugability.

[0160] DETAILED DESCRIPTION

[0161] Unless otherwise indicated, the terms used in the specification and claims have the following meanings.

[0162] The amino acid sequences of the present application contain the standard one-letter or three-letter codes for the twenty amino acids, and all amino acid residues in the present application are preferably in the L-form, unless otherwise specified.

[0163] "Naturally occurring 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).

[0164] "Non-natural amino acid" refers to an amino acid that is not naturally encoded or found in the genetic code of any organism. They can be, for example, purely synthetic compounds. Examples of non-natural amino acids include, but are not limited to, 2,8-diamino-(S)-octanoic acid, 2,7-diamino-(S)-heptanoic acid, L(-)-ornithine, L-2,4-diaminobutyric acid, hydroxyproline, gamma-carboxyglutamate, O-phosphoserine, azetidine carboxylic 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, 2,2'-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, C-terminal carboxyl groups, N-terminal amino groups, and / or side chain functional groups of natural amino acids or non-natural amino acids are chemically modified.

[0165] "Fatty acid" refers to a carboxylic acid having a long aliphatic tail (chain), which can be saturated or unsaturated; in the present invention, a fatty acid is a carboxylic acid having a C4-C30 straight chain or branched aliphatic group.

[0166] Other terms:

[0167] Polypeptide compounds can include amino acids having: amide bond hydrogen moieties replaced by methyl (N-methylation) or other alkyl groups, peptide bonds replaced with chemical groups or chemical linkages resistant to chemical or enzymatic treatment, N-terminal and C-terminal modifications.

[0168] Some polypeptides of the application can be cyclic. Cyclic polypeptides include any polypeptide having one or more cyclic features such as loops, bridging moieties, and / or internal linkages as part of its structure. As used herein, the term "bridging moiety" refers to one or more components of a bridge formed between two adjacent or non-adjacent amino acids, non-natural amino acids, or non-amino acid residues in a polypeptide. Bridging moieties can have any size or composition. In some embodiments, a bridging moiety can comprise one or more chemical bonds between two adjacent or non-adjacent amino acids, non-natural amino acids, non-amino acid residues, or combinations thereof. In some embodiments, such chemical bonds can be between one or more functional groups on adjacent or non-adjacent amino acids, non-natural amino acids, non-amino acid residues, or combinations thereof. Bridging moieties can comprise one or more features including, but not limited to, amide bonds (lactams), disulfide bonds, thioether bonds, aromatic rings, triazole rings, hydrocarbon chains, and -COCH2-N-CH2CO- (where N is a substituted or unsubstituted -NH-). In some embodiments, a bridging moiety comprises an amide bond between an amine functional group and a carboxylic acid functional group, each present in an amino acid, non-natural amino acid, or non-amino acid residue side chain. In some embodiments, the amine or carboxylic acid functional group is part of a non-amino acid residue or non-natural amino acid residue. In some cases, a bridging moiety can comprise a bond formed between residues including, but not limited to, lysine and aspartic acid. Bridging moieties can be formed using olefin metathesis via a cyclization reaction. In some embodiments, a bridging moiety comprises a disulfide bond formed between two thiol-containing residues, including but not limited to cysteine. In some embodiments, a bridging moiety comprises one or more thioether bonds. Such thioether bonds can include those found in cycloalkyl sulfide compounds. These bonds are formed during a chemical cyclization reaction between a chloroacetic N-terminal modification group and a cysteine residue. In some cases, a bridging moiety comprises one or more triazole rings. In some cases, a bridging moiety comprises a -COCH2-N-CH2CO- structure (where N is a substituted or unsubstituted -NH-), "substituted" as defined below, and in addition N can be substituted with a side chain as shown by Y2. In some embodiments, a bridging moiety comprises a non-protein or non-polypeptide based moiety including, but not limited to, a cyclic ring including, but not limited to, an aromatic ring structure (e.g., a xylyl group). Such bridging moieties can be introduced by reaction with a reagent containing multiple reactive halogens including, but not limited to, poly(bromomethyl)benzene, poly(bromomethyl)pyridine, poly(bromomethyl)alkylbenzene, and / or (E)-1,4-dibromo-but-2-ene.

[0169] A "modified amino acid residue" of the application refers to an amino acid residue that is modified by a side chain or substituted by a substituent, including but not limited to a structure shown by Y2 of the application.

[0170] Amino acid abbreviations and Chinese name correspondence table

[0171] The amino acid residues used in the polypeptide sequences of the present application use the amino acid shorthand.

[0172] Unless otherwise specifically noted, the amino acids described herein are "L-form amino acids," and "D(X)" indicates a "D-form amino acid," indicated by the substitution of the letter or three-letter amino acid code for the variable "X."

[0173] "[cyclo(x,y)]" refers to a bond formed between two amino acid residues, where the numerical identifiers x and y locate the positions of the residues involved in the bond.

[0174] The term "alkyl" refers to saturated aliphatic hydrocarbon groups that are straight-chain or branched-chain groups comprising 1 to 30 carbon atoms, preferably alkyl groups containing 10 to 28 carbon atoms, more preferably 12 to 28 carbon atoms, most preferably 16 to 24 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-tridecyl, n-pentadecyl, n-heptadecyl, n-nonadecyl, n-undecadecyl, n-tricosyl, n-pentacosyl, n-heptacosyl, n-nonacosyl, n- hentriacontyl, and the like, as well as various branched isomers thereof. More preferred are long chain alkyl groups containing 12 to 28 carbon atoms, non-limiting examples of which include n-tridecyl, n-pentadecyl, n-heptadecyl, n-nonadecyl, n-undecadecyl, n-tricosyl, n-pentacosyl, n-heptacosyl, n-nonacosyl, or n-hentriacontyl.

[0175] The term "cycloalkyl" refers to saturated or partially unsaturated aliphatic hydrocarbon monocyclic, polycyclic (two or more) cyclic groups, which can be optionally substituted with one or more substituents. In particular embodiments, the cycloalkyl ring comprises 3 to 20 (C 3-20 ), 3 to 12 (C 3-12 ), 3 to 8 (C 3-8 ), or 3 to 6 (C 3-6 ) carbon atoms; in one embodiment, the cycloalkyl ring comprises 6 to 14 (C 6-14 ) or 7 to 10 (C 7-10) atoms; it can contain one or more double bonds but does not have a completely conjugated pi-electron system. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, or cyclooctyl, and the like; polycyclic cycloalkyl groups include spirocycloalkyl, fused cycloalkyl, and bridged cycloalkyl groups. In one embodiment, the cycloalkyl group is an optionally substituted cycloalkyl group described elsewhere herein or a cycloalkyl group optionally fused to a heterocyclyl, aryl, or heteroaryl group, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptyl, and the like.

[0176] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic ring-containing hydrocarbon radical in which one or more of the ring atoms is a heteroatom selected from nitrogen, oxygen, boron, phosphorus, or sulfur, wherein the nitrogen, phosphorus, or sulfur atom is optionally oxidized, the nitrogen atom is optionally quaternized, the ring carbon atoms are optionally substituted with oxygen but excluding -O-O-, -O-S- ring moieties, and the remaining ring atoms are carbon, which can contain one or more double bonds but does not have a completely conjugated pi-electron system. In particular embodiments, the heterocyclyl group contains 3 to 20, 3 to 12, 3 to 8, or 3 to 6 ring atoms, of which 1 to 4 are heteroatoms; in one embodiment, the heterocyclyl group contains 3 to 6, 4 to 6, 3 to 8, 3 to 10, 6 to 10, or 7 to 11 ring atoms; in one embodiment, the heterocyclyl group contains 3 to 8 (e.g., 3, 4, 5, 6, 7, 8) ring atoms. Non-limiting examples of monocyclic heterocyclyl groups include tetrahydropyrrolyl, azetidinyl, oxetanyl, oxepanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and pyranyl, and the like. Polycyclic heterocyclyl groups include spiroheterocyclyl, fused heterocyclyl, and bridged heterocyclyl groups. In one embodiment, the heterocyclyl group is an optionally substituted heterocyclyl group described elsewhere herein or a heterocyclyl group further annelated to other cycloalkyl, heterocyclyl, aryl, and heteroaryl groups through any two or more atoms on the ring.

[0177] The term "aryl" refers to all-carbon monocyclic or fused polycyclic (that is, rings which share adjacent pairs of carbon atoms) rings having a conjugated pi-electron system, which can be optionally substituted by one or more substituents. In particular embodiments, aryl groups contain from 6 to 20, from 6 to 14, or from 6 to 10 ring atoms; in one embodiment, aryl can further refer to a bicyclic, tricyclic, or tetracyclic ring system, wherein at least one ring is aromatic, and the other rings can be saturated, partially unsaturated, carbocyclic, or a ring containing one or more heteroatoms independently selected from O, S, and N; in one embodiment, the aryl group is selected from benzo 5-10 membered heteroaryl, benzo 3-10 membered cycloalkyl, or benzo 3-10 membered heterocyclyl. In one embodiment, the aryl group is selected from benzo 5-6 membered heteroaryl, benzo 3-6 membered cycloalkyl, or benzo 3-6 membered heterocyclyl, wherein the heterocyclyl is a heterocyclyl containing 1-3 nitrogen atoms, oxygen atoms, or sulfur atoms. Non-limiting examples include phenyl, naphthyl, fluorenyl, azulenyl, anthracenyl, phenanthrenyl, pyrenyl, biphenyl, terphenyl, dihydronaphthyl, indenyl, tetrahydronaphthyl (tetrahydroanthracenyl).

[0178] The term "heteroaryl" refers to an optionally substituted monocyclic, polycyclic group or ring system comprising at least one aromatic ring having one or more heteroatoms independently selected from O, S, and N. In particular embodiments, the heteroaryl group comprises 5 to 20, 5 to 14, or 5 to 10 ring atoms, of which 1 to 4 are heteroatoms; in one embodiment, the heteroaryl group comprises 5 or 6 ring atoms; in particular embodiments, the heteroaryl group can further refer to a bicyclic, tricyclic, or tetracyclic ring system, wherein at least one ring is an aromatic ring having one or more heteroatoms independently selected from O, S, and N, and the other rings can be saturated, partially unsaturated carbocyclic rings or rings comprising one or more heteroatoms independently selected from O, S, and N. In one embodiment, the heteroaryl group is selected from heteroaryl and 6-10 membered aryl, heteroaryl and 3-10 membered cycloalkyl, or heteroaryl and 3-10 membered heterocyclyl; in a further embodiment, the heteroaryl group is selected from 5- or 6-membered heteroaryl and 6-10 membered aryl, 5- or 6-membered heteroaryl and 3-6 membered cycloalkyl, 5- or 6-membered heteroaryl and 3-6 membered heterocyclyl, wherein the heterocyclyl is a heterocyclyl comprising 1-3 nitrogen atoms, oxygen atoms, or sulfur atoms. Non-limiting examples include furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl, benzofuranyl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothienyl, benzothiophenyl, benzothiophenyl, benzotriazolyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolyl, naphthridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridinyl, pyrrolopyridinyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidinyl, thienopyridinyl, acridinyl, benzoindolyl, carbazolyl, diphenofuranyl, phenanthrolinyl, phenanthridinyl, phenarsenazinyl, phenoxazinyl, phenothiazinyl, phenoxazinyl, xanthenyl.

[0179] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein alkyl or cycloalkyl are as defined above. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, or cyclohexoxy. In one embodiment, the alkoxy group is an optionally substituted alkoxy group as described elsewhere herein.

[0180] The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above. Non-limiting examples of haloalkyl groups include trifluoromethyl.

[0181] The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group, wherein alkyl is as defined above.

[0182] The term "substituted" means that any one or more hydrogen atoms on a given atom are replaced with a substituent, provided that the valence of the given atom is properly satisfied and that the resulting compound is stable. In one embodiment, when the substituent is oxo (i.e., =0), it means that two hydrogen atoms are replaced. The term "optionally substituted" means that the group can or can not be substituted, and unless otherwise specified, the types and number of substituents are any that are chemically possible. It goes without saying that the substituents are only in their possible chemical positions, which can be determined (experimentally or theoretically) by one of skill in the art without undue effort, as to what is possible or impossible. For example, an amino or hydroxyl group with a free hydrogen can not be stable when bound to a carbon atom with an unsaturation (e.g., olefinic). The "substituents" include hydrogen, halogen, amino, nitro, hydroxyl, cyano, thiol, oxo, thioxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1- 6hydroxyalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-14 aryl, and 5-14 membered heteroaryl.

[0183] The term "halogen" means fluorine, chlorine, bromine, or iodine. The term "formyl" means -CHO. "TFA" means trifluoroacetic acid.

[0184] The term "acetyl" means -C(O)CH3. "DCM" means dichloromethane. "HFIP" means hexafluoroisopropanol.

[0185] "MeOH" means methanol. "DMF" means N,N-dimethylformamide.

[0186] "ACN" means acetonitrile. "PEG" means polyethylene glycol. "DIEA" means diisopropylethylamine.

[0187] "HATU" means 2-(7-oxabenzo[l]oxazol-3-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0188] "HBTU" means benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0189] "HCTU" means 6-chlorobenzotriazol-1,1,3,3-tetramethyluronium hexafluorophosphate.

[0190] "DMAP" means 4-dimethylaminopyridine. "PE" means petroleum ether. "EA" means ethyl acetate.

[0191] "TLC" refers to thin layer chromatography. "Boc group" refers to tert-butyloxycarbonyl.

[0192] "Fmoc group" refers to fluorenylmethyloxycarbonyl. "Tis" refers to triisopropylsilane.

[0193] "Dde" refers to l-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl. "DTT" refers to dithiothreitol.

[0194] "Alloc" refers to aminoallyloxycarbonyl. "Mtt" refers to 4-methyltrityl.

[0195] "DEPBT" refers to 3-(diethoxyphosphoryloxy)-l,2,3-benzotriazin-4-one.

[0196] "OAI1" refers to carboxyallyl ester.

[0197] "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 different phrases all express the same meaning, i.e., X can be any one or several of A, B, and C.

[0198] The hydrogen atoms described in the present application can be replaced by its isotope deuterium, and any hydrogen atom in the compounds of the embodiments described in the present application can also be replaced by deuterium.

[0199] "Pharmaceutical composition" means a mixture of one or more polypeptides described in the present application and derivatives thereof, 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 to an organism and to facilitate absorption of the active ingredient to thereby exert a biological activity.

[0200] "Pharmaceutically acceptable salt" refers to a salt of the polypeptides of the present application, which is safe and effective when used in a mammal, and has the appropriate biological activity.

[0201] In the present application, the term "treatment" includes inhibition, slowing, stopping, or reversing the progression or severity of an existing symptom or disease. DETAILED DESCRIPTION

[0202] In order to more specifically explain the present application, the following specific embodiments are provided in the present specification, but the present application is not limited to only these embodiments.

[0203] 1. Experimental reagents

[0204] 2. Experimental instruments

[0205] 3. Specific embodiments

[0206] 3.1 Synthesis of intermediates

[0207] Intermediate L1

[0208] First step: Weigh 2-CTC Resin (346.4 g, Substitution 1.5 mmol / g) into the solid phase reactor, add anhydrous DCM (2 L), nitrogen bubbling for 30 minutes for swelling, and dry; weigh L1-P1 (100 g, 259.7 mmol) and DIEA (180.5 mL, 1.04 mol) into anhydrous DCM (2 L), and add into the solid phase reactor, nitrogen bubbling for 5 hours, and dry; add a mixture of DIEA (180.5 mL, 1.04 mol) in MeOH / DCM (1:4, 2 L), and bubble for 1.5 hours, and dry; wash with DMF (2 L) and IPA (2 L) alternately for two times (nitrogen bubbling for 10 minutes), and then wash with DMF (2 L) for three times, and dry to obtain L1-1, which is directly used for the next step.

[0209] Second step: add 20% piperidine / DMF solution (2 L*2) for 5 minutes respectively, and dry after 20 minutes; wash the resin with DMF (2 L) for two times (nitrogen bubbling for 10 minutes), and dry to obtain L1-2, which shows deep blue color in ninhydrin test.

[0210] Third step: weigh L1-P1 (150 g, 389.6 mmol), HATU (148.05 g, 389.6 mmol) and DIEA (180.5 mL, 1.04 mol) into DMF (2 L), and add into the solid phase reactor, nitrogen bubbling for 2 hours, and dry, which shows light yellow color in ninhydrin test; wash with DMF (2 L) and IPA (2 L) alternately for two times (nitrogen bubbling for 10 minutes), and then wash with DMF (2 L) for three times, and dry to obtain L1-3, which is directly used for the next step.

[0211] Fourth step: add 20% piperidine / DMF solution (2 L*2) for 5 minutes respectively, and dry after 20 minutes; wash the resin with DMF (2 L) for two times (nitrogen bubbling for 10 minutes), and dry to obtain L1-4, which shows deep blue color in ninhydrin test.

[0212] Step 5: Weigh L1-P2 (165.58 g, 389.6 mmol), HATU (148.05 g, 389.6 mmol) and DIEA (180.5 mL, 1.04 mol) into DMF (2 L) and add to the solid phase reactor. Bubble nitrogen for 2 hours, drain and dry, and test with ninhydrin to show a light yellow color. Wash with DMF (2 L) and IPA (2 L) alternately for two times (bubble nitrogen for 10 minutes), and then wash with DMF (2 L) for three times. Drain and dry to get L1-5, which is used directly in the next step.

[0213] Step 6: Add 20% piperidine / DMF solution (2 L*2) and bubble for 5 minutes, respectively. Drain and dry after 20 minutes. Wash the resin with DMF (2 L) for two times (bubble nitrogen for 10 minutes), and then drain and dry to get L1-6, which shows a dark blue color in the ninhydrin test.

[0214] Step 7: Weigh L1-P3 (133.25 g, 389.6 mmol), HATU (148.05 g, 389.6 mmol) and DIEA (180.5 mL, 1.04 mol) into DMF (2 L) and add to the solid phase reactor. Bubble nitrogen for 2 hours, drain and dry, and test with ninhydrin to show a light yellow color. Wash with DMF (2 L) and IPA (2 L) alternately for two times (bubble nitrogen for 10 minutes), and then wash with DMF (2 L) for three times. Drain and dry to get L1-7.

[0215] Step 8: Wash with isopropyl ether (2 L*4) (bubble nitrogen for 10 minutes), drain and dry. Then transfer the resin to a 5 L three-necked flask, add 20% hexafluoroisopropanol / DCM solution (2 L), and stir overnight at room temperature. Drain and filter, and then stir the resin again with 20% hexafluoroisopropanol / DCM solution (2 L) for 3 hours. Drain and filter, combine the filtrates, and concentrate the residue on a silica gel column (MeOH:DCM = 0-15% gradient elution) to get compound L1-8 (132 g). LC / MS: [M+H] = 818.6. +

[0216] 1 H NMR (400 MHz, CD3OD) δ 4.26 (dd, 1H), 4.15 (s, 2H), 4.03 (s, 2H), 3.74-3.70 (m, 4H), 3.69-3.66 (m, 4H), 3.62-3.56 (m, 4H), 3.47 (t, 2H), 3.40 (t, 2H), 2.37-2.18 (m, 6H), 2.17-2.05 (m, 1H), 1.98-1.86 (m, 1H), 1.69-1.55 (m, 4H), 1.47 (d, 18H), 1.32 (s, 20H).

[0217] ​Ninth step: To a solution of compound L1-9 (60 g, 73.35 mmol) in ACN (600 mL) was added DIEA (47.40 g, 366.73 mmol) and HATU (35.97 g, 94.61 mmol) at 0 °C, then compound L1-P4 (23.18 g, 88.01 mmol) was added, and the mixture was stirred at 0 °C for 1 h. The reaction was concentrated under reduced pressure, diluted with water, and extracted with ethyl acetate twice. The organic phase was dried, filtered, and concentrated to give compound L1-9 (95.3 g, crude). LC / MS: [M] + = 1062.

[0218] Tenth step: To a solution of compound L1-9 (15 g, 14.12 mmol) in THF (100 mL) was added a solution of LiOH (1.78 g, 42.37 mmol) in water (50 mL) at 0 °C, and the mixture was stirred at 0 °C for 1 h. The reaction was concentrated, and the residue was adjusted to pH 7 with 2N HCl solution, and purified by reverse phase C18 column chromatography (5% - 40%, 40% - 70%, ACN / 0.03% TFA / water) to give intermediate L1 (10.09 g, TFA salt). LC / MS: [M] + = 1048.7.

[0219] 1 HNMR (400 MHz, CD3OD) δ 4.25 (dd, 1H), 4.04 (s, 2H), 4.01 (s, 2H), 3.95 (s, 2H), 3.74 (t, 4H), 3.72 - 3.63 (m, 14H), 3.62 - 3.54 (m, 6H), 3.47 (t, 2H), 3.39 (d, 2H), 3.23 (s, 6H), 2.55 (t, 2H), 2.32 - 2.19 (m, 6H), 2.15 - 2.06 (m, 1H), 1.90 (dd, 1H), 1.69 - 1.51 (m, 4H), 1.46 (d, 18H), 1.30 (s, 20H).

[0220] Intermediate L2

[0221] Intermediate L2 was prepared according to the procedure described for the synthesis of intermediate L1. LC / MS: [M] + = 1104.6.

[0222] 1HNMR (400 MHz, CD3OD) δ 4.26 (dd, 1H), 4.04 (d, 4H), 3.97 (d, 2H), 3.80-3.73 (m, 4H), 3.73-3.54 (m, 20H), 3.48 (t, 2H), 3.42-3.35 (m, 2H), 3.24 (s, 6H), 2.57 (t, 2H), 2.34-2.19 (m, 6H), 2.18-2.07 (m, 1H), 1.92 (ddd, 1H), 1.68-1.55 (m, 4H), 1.47 (d, 18H), 1.31 (s, 28H).

[0223] The intermediate L2 can also be prepared according to the following synthesis method:

[0224] First step: L1-P4-1 (100 g, 414.80 mmol) was dissolved in DMF (1000 mL), NaHCO3 (139.38 g, 1.66 mol) was added, and iodomethane (235.51 g, 1.66 mmol) was slowly added dropwise, and the reaction was carried out at room temperature overnight. The reaction solution was quenched by adding 3 L of water, extracted with ethyl acetate (1 L*3), and the organic phase was washed with saturated brine (1.5 L*2), and the organic phase was dried and concentrated to obtain compound L1-P4-2 (96 g), yield: 90.7%. LC / MS: [M+H] + = 256.

[0225] Second step: Compound L1-P4-2 (96 g, 376.3 mmol) was dissolved in acetonitrile (1000 mL), and tert-butyl N-[2-(dimethylamino)ethyl]carbamate (141.69 g, 752.63 mmol) was added, and the reaction was carried out at 50°C overnight. The reaction solution was directly spin-dried, added to 1 L of water, washed with ethyl acetate (600 mL*3), and the aqueous phase was freeze-dried to obtain compound L1-P4-3 (99 g), yield: 72.4%. LC / MS: [M+H] + = 363.

[0226] Third step: Compound L1-P4-3 (99 g, 272.38 mmol) was dissolved in TFA (500 mL), and the reaction was carried out at room temperature for 2 hours. Concentration under reduced pressure, the crude product was adjusted to pH 9-10 with DIEA under ice bath, and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound L1-P4 (56.32 g), yield: 78.5%. LC / MS: [M+H] + = 263.

[0227] Fourth step: Dissolve L1-6 (155 g, 388.84 mmol) and HATU (148 g, 392.29 mmol) in DMF (2000 ml), add to the solid-phase synthesis reactor; add DIEA (134 g, 1.04 mol), N2 bubble for 3 hours, and dry. Wash with DMF (2 L) and IPA (2 L) alternately twice, then wash with DMF (2 L) for 3 times, and isopropyl ether (2 L*4), and dry to obtain compound L2-1, which can be directly used in the next step.

[0228] Fifth step: Add the resin to a 5000 ml single-neck flask, add 20% hexafluoroisopropanol / DCM solution (2 L), and stir at room temperature overnight; filter, and collect the filtrate. Add the filter cake to a 5000 ml single-neck flask again, add 20% hexafluoroisopropanol / DCM solution (2 L), and stir for 8 hours. Filter, combine all the filtrates, concentrate the residue under reduced pressure, and purify the residue by silica gel column chromatography (DCM / MeOH system) to obtain compound L2-2 (150.0 g). LC / MS: [M+H] + = 875;

[0229] 1 H NMR (400 MHz, CD3OD) δ 4.26 (dd, 1H), 4.15 (s, 2H), 4.03 (s, 2H), 3.73-3.66 (m, 8H), 3.59 (dt, 4H), 3.47 (t, 2H), 3.39 (dd, 2H), 2.36-2.19 (m, 6H), 2.18-2.06 (m, 1H), 1.99-1.84 (m, 1H), 1.67-1.54 (m, 4H), 1.47 (d, 18H), 1.31 (s, 28H).

[0230] Sixth step: Dissolve compound L2-2 (68 g, 77.8 mmol), HATU (44 g, 115.7 mmol), and DIEA (40.2 g, 311.3 mmol) in acetonitrile (800 ml), add a solution of L1-P4 (38.2 g) in acetonitrile (50 mL) to the reaction liquid under ice bath stirring, and keep stirring at 0°C for 1 hour. Concentrate the reaction liquid under reduced pressure, and purify the residue by C18 reverse column chromatography (5%-40%, 40%-70%, ACN / 0.03% TFA / water) to obtain compound L2-3. LC / MS: [M] + = 1119.6;

[0231] 1H NMR (400 MHz, CD3OD) δ 4.26 (dd, 1H), 4.04 (d, 4H), 3.97 (d, 2H), 3.80-3.73 (m, 4H), 3.73-3.54 (m, 20H), 3.48 (t, 2H), 3.42-3.35 (m, 2H), 3.24 (s, 6H), 2.57 (t, 2H), 2.34-2.19 (m, 6H), 2.18-2.07 (m, 1H), 1.92 (ddd, 1H), 1.68-1.55 (m, 4H), 1.47 (d, 18H), 1.31 (s, 28H).

[0232] Step 7: Compound L2-3 (56 g, 50 mmol) was dissolved in THF / H2O (1:1, 600 mL), LiOH (6.3 g, 150 mmol) was added at 0 °C, and the mixture was stirred at 0 °C for 1 h. The pH of the reaction solution was adjusted to 6-7 with 2 M dilute hydrochloric acid, and the residue was concentrated under reduced pressure and purified by C18 reverse column chromatography (5%-40%, 40%-70%, ACN / 0.03% TFA / water) to give intermediate L2 (27 g). LC / MS: [M] + = 1104.6.

[0233] 1 H NMR (400 MHz, CD3OD) δ 4.26 (dd, 1H), 4.04 (d, 4H), 3.97 (d, 2H), 3.80-3.73 (m, 4H), 3.73-3.54 (m, 20H), 3.48 (t, 2H), 3.42-3.35 (m, 2H), 3.24 (s, 6H), 2.57 (t, 2H), 2.34-2.19 (m, 6H), 2.18-2.07 (m, 1H), 1.92 (ddd, 1H), 1.68-1.55 (m, 4H), 1.47 (d, 18H), 1.31 (s, 28H).

[0234] The following intermediates were prepared according to the synthesis method of intermediate L1:

[0235] Intermediate L3

[0236] LC / MS: [M] + = 959.6.

[0237] 1H NMR (400 MHz, CD3OD) δ 4.27 (s, 1H), 4.06 (s, 2H), 3.97 (s, 2H), 3.83 - 3.63 (m, 14H), 3.62 - 3.53 (m, 4H), 3.40 (td, 2H), 3.24 (s, 6H), 2.57 (t, 2H), 2.36 - 2.19 (m, 6H), 2.19 - 2.06 (m, 1H), 1.91 (ddd, 1H), 1.69 - 1.54 (m, 4H), 1.47 (d, 18H), 1.31 (s, 28H).

[0238] Intermediate L4

[0239] LC / MS: [M] + = 1077.2.

[0240] 1 H NMR (400 MHz, CD3OD) δ 4.27 (dd, 1H), 4.06 (s, 2H), 4.03 (s, 2H), 3.97 (s, 2H), 3.76 (t, 4H), 3.73 - 3.64 (m, 14H), 3.63 - 3.55 (m, 6H), 3.48 (t, 2H), 3.42 - 3.37 (m, 2H), 3.24 (s, 6H), 2.57 (t, 2H), 2.35 - 2.20 (m, 6H), 2.19 - 2.06 (m, 1H), 1.98 - 1.82 (m, 1H), 1.69 - 1.54 (m, 4H), 1.48 (d, 18H), 1.31 (s, 24H).

[0241] Intermediate L6

[0242] LC / MS: [(M+H) / 2] + = 625.1;

[0243] 1 H NMR (400 MHz, CD3OD) δ 4.33 - 4.19 (m, 1H), 4.04 (d, 6H), 3.97 (d, 2H), 3.76 (t, 4H), 3.73 - 3.64 (m, 18H), 3.64 - 3.55 (m, 8H), 3.49 (t, 4H), 3.40 (d, 2H), 3.24 (s, 6H), 2.57 (t, 2H), 2.31 (t, 2H), 2.28 - 2.19 (m, 4H), 2.18 - 2.08 (m, 1H), 1.97 - 1.86 (m, 1H), 1.66 - 1.54 (m, 4H), 1.48 (d, 18H), 1.31 (s, 28H).

[0244] intermediate L7

[0245] LC / MS: [(M+H) / 2] + =580.9.

[0246] intermediate P1

[0247] Intermediate P1 can be prepared by referring to WO2023 / 141044 and the synthesis method of intermediate L1.

[0248] LC / MS: [(M+2H) / 2] + =517.8.

[0249] intermediate P2

[0250] Intermediate P2 can be prepared by referring to WO2023 / 141044 and the synthesis method of intermediate L1.

[0251] LC / MS: [(M+H) / 2] + =632.9.

[0252] Referring to the synthesis method of intermediate L1, by replacing the starting material L1-P3 with 19-(bis(benzyloxy)phosphoryl)nonadecanoic acid (CAS: 2807449-20-1), intermediate P2 can be prepared from L1-6.

[0253] Intermediate A1

[0254] Step 1: 1-Cyclopropylacetone (5 g, 50.9 mmol) was dissolved in a methanol solution of ammonia (30 mL), and stirred at 0 °C for 3 hours. Then, trimethylcyanosilane (5.05 g, 50.9 mmol) was added dropwise at 0 °C, and the mixture was gradually brought to room temperature and stirred for 12 hours. The reaction solution was diluted with water (100 mL), extracted with ethyl acetate (50 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 2-amino-3-cyclopropyl-2-methylpropionitrile A1-1 (4.2 g), yield: 66.4%. LC / MS: [M + H] + =125.1.

[0255] 1 HNMR(400MHz, DMSO-d6)δ2.53(s,2H),1.52(dd,1H),1.43(dd,1H),0.89–0.76(m,1H),0.53–0.43(m,2H),0.15(m,2H).

[0256] Second Step: 2-amino-3-cyclopropyl-2-methylpropionitrile A1-1 (2.17 g, 17.5 mmol) was dissolved in methanol (60 mL), and hydrochloric acid gas was continuously bubbled in at -5 ~ 0 °C for 6 hours. After the reaction solution was concentrated under reduced pressure to remove most of the solvent, saturated sodium bicarbonate solution was added to adjust the pH to 9, and dichloromethane (30 mL*4) was extracted, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain methyl 2-amino-3-cyclopropyl-2-methylpropanoate A1-2 (2.25 g), with a yield of 81.9%. LC / MS: [M+H] + = 158.1.

[0257] Third Step: Methyl 2-amino-3-cyclopropyl-2-methylpropanoate A1-2 (2.2 g, 14.0 mmol) was dissolved in dioxane (30 mL) and water (30 mL), and then lithium hydroxide monohydrate (1.20 g, 28.0 mmol) was added, and stirred at room temperature for 2 hours. The reaction solution was used directly in the next step without any treatment. LC / MS: [M+H] + = 144.1.

[0258] Fourth Step: Sodium carbonate (888 mg, 8.40 mmol) was added to the reaction solution of the previous step, and chloroformic acid-9-fluorenylmethyl ester (1.80 g, 6.96 mmol) was added at 0 °C, and gradually stirred at room temperature for 1 hour. Then sodium carbonate (888 mg, 8.4 mmol) and chloroformic acid-9-fluorenylmethyl ester (0.9 g, 3.48 mmol) were added at 0 °C, and stirred at room temperature for 16 hours. The reaction solution was added with 1M dilute hydrochloric acid solution in an ice bath, adjusted to pH 2, diluted with water (100 mL), extracted with dichloromethane (100 mL*2), washed with water and saturated sodium chloride brine, respectively, dried over anhydrous sodium sulfate, filtered, and the residue was concentrated under reduced pressure and purified by preparative HPLC (formic acid system) to obtain 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropyl-2-methylpropanoic acid (1.6 g). Further purification by chiral HPLC (CHIRALPAK-IG-H / Hexane:EtOH=70%:30%-0.1%FA, RT=9.00 min) to obtain intermediate A1 (Fmoc-α-Me-cpAla-OH, 670 mg), with a yield of 13%.

[0259] MS m / z (ESI): 366.2 [M+H] + .

[0260] 1H NMR (400 MHz, DMSO-d6) δ 12.40 (s, 1H), 7.89 (d, 2H), 7.72 (d, 2H), 7.45 - 7.38 (m, 3H), 7.33 (td, 2H), 4.31 - 4.16 (m, 3H), 1.78 (dd, 1H), 1.59 (dd, 1H), 1.40 (s, 3H), 0.62 (s, 1H), 0.37 (d, 2H), 0.05 - -0.01 (m, 2H).

[0261] 3.2 Synthesis of the examples

[0262] 3.2.1 Chemical synthesis of compound no. 13 H-Aib-H-G-T-F-T-S-D-Y-S-I-Achx-L-E-K(-NEG-OEG-OEG-γGlu-C20diacid)-K-A-A-Q-E-F-V-E-W-L-L-A- G-G-P-S-S-G-A-P-P-P-S-NH2

[0263] 3.2.1.1 Coupling of Fmoc-Ser(tBu)-OH to Rink Amide-AM Resin

[0264] Weigh Rink Amide-AM Resin (1 mmol) into the reactor, add DMF (20 mL) to the reactor and nitrogen bubbling to swell for 0.5 hours, and then dry the solvent.

[0265] Add 20% piperidine / DMF (20 mL) to the reactor, and nitrogen bubbling for 5 minutes, and then dry the solvent. Add 20% piperidine / DMF (20 mL) to the reactor, and nitrogen bubbling for 20 minutes, and then dry the solvent. Wash with DMF (20 mL) for 4 times, 2 minutes each time, and then dry the solvent. Ninhydrin test, the resin is blue.

[0266] Weigh Fmoc-Ser(tBu)-OH (3.0 eq), HOBt (3.0 eq), DIC (3.0 eq) into DMF (20 mL), mix well, and then add to the reactor, and nitrogen bubbling for 3 hours. Ninhydrin test, the resin does not change blue. Dry the reaction solution, and then wash with DMF, isopropyl alcohol, DMF, isopropyl alcohol, DMF, DMF, and DMF.

[0267] 3.2.1.2 Coupling of the peptide chain sequence

[0268] According to the sequence of the peptide chain of the compound, the sequence of the peptide chain is N-terminal to C-terminal (H-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Achx-Leu-Glu-Lys-Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2). The solid-phase synthesis is synthesized in the order of C-terminal to N-terminal. The amount of the protected amino acid and the condensation reagent and the condensation method thereof are the same as those of the coupling of Fmoc-Ser(tBu)-OH to Rink Amide-AM Resin. The protected amino acids used in the synthesis process are Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH, Fmoc-Ala-OH, Fmoc-Lys(Boc)-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Achx-OH, Fmoc-Ile-OH, Fmoc-Ser(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-His(Trt)-Gly-OH, Boc-His(Trt)-Aib-OH, respectively. The coupling of the protected amino acid and the deprotection of Fmoc are repeated to obtain a linear resin peptide.

[0269] 3.2.1.3. Removal of the Alloc protecting group from the linear resin peptide

[0270] Add 20 mL of DCM to the reactor, bubble under nitrogen for 5 minutes, and then remove the solvent. Repeat this step once more. Weigh 10 eq of morpholine and 0.2 eq of Pd(PPh3)4, dissolve them in 20 mL of DCM, add them to the reactor, bubble under nitrogen for 20 minutes, and then remove the reaction solution. Repeat this step twice more. Wash three times with 20 mL of DCM, 2 minutes each time, and then remove the solvent. Wash three times with 20 mL of DMF, 2 minutes each time, and then remove the solvent. The resin will turn blue when tested for ninhydrin.

[0271] 3.2.1.4 Side chain coupling

[0272] Weigh intermediate L2 (3.0 eq), HCTU (3.0 eq), and DIEA (6.0 eq) and add them to DMF (20 mL). Mix well and then add to the reactor. React under nitrogen bubbling for 24 hours. The resin does not turn blue when tested with ninhydrin. Dry the reaction solution and wash sequentially with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF.

[0273] 3.2.1.5 Cleavage of Resin Peptides

[0274] Prepare 40 mL of lysis buffer according to the ratio of TFA:DTT:Tis:H2O = 92.5:2.5:2.5:2.5. Add the resin peptide to the lysis buffer and stir at room temperature for 2 hours. After the reaction is complete, filter, and flush the filtrate into isopropyl ether (400 mL), filter again to obtain the crude peptide.

[0275] 3.2.1.7 Purification of crude peptides

[0276] The crude peptide was dissolved in a 20% acetonitrile / water solution, filtered through a 0.45 μm membrane, and then separated using a reversed-phase high-performance liquid chromatography (RP-HPLC) system. The mobile phases for purification were A (0.1% trifluoroacetic acid aqueous solution) and B (acetonitrile). A C18 reversed-phase column was used, with the chromatographic detection wavelength set at 220 nm and the flow rate at 70 mL / min. The target fraction was collected, concentrated using the reversed-phase column, and then lyophilized. The purity of the sample was determined by HPLC to be 95.7%; LC / MS (ESI): [(M+3H) / 4] + =1275.00,[(M+4H) / 5] + =1020.20, consistent with the compound's molecular weight of 5097.9.

[0277] 3.2.2 Chemical Synthesis of Compound No. 15 H-Aib-H-G-T-F-T-S-D-Y-S-I- a-Me-cpA-L-E-K(-NEG-OEG-OEG- yGlu-C20diacid)-K-A-A-Q-E-F-V-E-W-L-L-A-G-G-P-S-S-G-A-P-P-P-S-NH2

[0278] 3.2.2.1 Coupling of Fmoc-Ser(tBu)-OH to Rink Amide-AM Resin

[0279] Weigh Rink Amide-AM Resin (1 mmol) into the reactor, add DMF (20 mL) to the reactor and nitrogen bubble to swell for 0.5 hour, then dry.

[0280] Add 20% piperidine / DMF (20 mL) to the reactor, nitrogen bubble for 5 minutes, then dry. Add 20% piperidine / DMF (20 mL) to the reactor, nitrogen bubble for 20 minutes, then dry. Wash with DMF (20 mL) for 4 times, 2 minutes each time, then dry. Ninhydrin test, the resin is blue.

[0281] Weigh Fmoc-Ser(tBu)-OH (3.0 eq), HOBt (3.0 eq), DIC (3.0 eq) into DMF (20 mL), mix well and add to the reactor, then nitrogen bubble for 2 hours. Ninhydrin test, the resin is not blue. Wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, DMF in turn.

[0282] 3.2.2.2 Coupling of the peptide chain sequence

[0283] According to the sequence of the peptide chain of the compound, the sequence of the peptide chain is N-terminal to C-terminal (H-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-α-Me-cpAla-Leu-Glu-Lys-Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2). The solid-phase synthesis is synthesized in the order of C-terminal to N-terminal. The amount of the protected amino acid and the condensation reagent and the condensation method thereof are the same as those of the coupling of Fmoc-Ser(tBu)-OH to Rink Amide-AM Resin. The protected amino acids used in the synthesis process are Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-α-Me-cpAla-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH, Fmoc-Ala-OH, Fmoc-Lys(Boc)-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Ser(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-His(Trt)-Gly-OH, Boc-His(Trt)-Aib-OH, successively. The coupling of the protected amino acid and the deprotection of Fmoc are repeated to complete the straight-chain resin peptide.

[0284] 3.2.2.3 Removal of the Alloc protecting group from the straight-chain resin peptide

[0285] Into the reactor, DCM (20 mL) was added, and nitrogen was bubbled for 2 minutes, and then the solution was drained. This step was repeated once more. Morpholine (10 eq) and Pd(PPh3)4(0.2 eq) were dissolved in DCM (20 mL) and added to the reactor, and nitrogen was bubbled for 20 minutes, and then the solution was drained. This step was repeated twice. DCM (20 mL) was added and washed for 2 minutes each time, and then the solution was drained. DMF (20 mL) was added and washed for 2 minutes each time, and then the solution was drained. Ninhydrin test, resin blue.

[0286] 3.2.2.4 Side chain coupling

[0287] Into the reactor, intermediate L2 (3.0 eq), HCTU (3.0 eq), and DIEA (6.0 eq) were added, and nitrogen was bubbled for 4 hours. Ninhydrin test, resin did not turn blue. The resin was washed with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF, in that order.

[0288] 3.2.2.5 Cleavage of the resin peptide

[0289] A 50 mL cleavage solution was prepared according to the ratio of TFA: DTT: Tis: H2O = 92.5: 2.5: 2.5: 2.5. The resin peptide was added to the cleavage solution, and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, the solution was filtered, and the filtrate was eluted into isopropyl ether (500 mL), and then filtered to obtain the crude peptide.

[0290] 3.2.2.6 Purification of the crude peptide

[0291] The crude peptide was dissolved in a 20% acetonitrile / water solution, filtered through a 0.45 um membrane, and then separated using a reverse phase high performance liquid chromatography system, with buffer A (0.1% trifluoroacetic acid in water) and buffer B (acetonitrile). The chromatographic column was a C18 reverse phase chromatographic column, the detection wavelength of the chromatograph was set to 220 nm during the purification process, and the flow rate was 70 mL / min. The target component was collected, and after reverse column concentration, concentration and freeze-drying. The sample was determined for purity by HPLC, 90%; LC / MS (ESI): [(M+3H) / 4] + = 1275.00, [(M+4H) / 5] + = 1020.20, which is consistent with the molecular weight of the compound 5097.9.

[0292] The compounds of the present application numbered 1-12, 14, 16-27, and 52-60, 65-82 can be synthesized according to the test scheme of the above examples.

[0293] Some of the compounds of the present application were detected for purity and confirmed structure by analytical UPLC and LC / MS as shown in the following table:

[0294] 3.2.3 Chemical synthesis of compound No. 28 cyclo(17,20)H-Aib-H-G-T-F-T-S-D-Y-S-I-L-L-E-K-K-A-A-K(-COCH2N(-OEG-OEG- γGlu-C20diacid)-CH2CO)-E-F-V-E-W-L-L-A-G-G-P-S-S-G-A-P-P-P-S-NH2

[0295] 3.2.3.1 Coupling of Fmoc-Ser(tBu)-OH with Rink Amide-AM Resin

[0296] Weigh Rink Amide-AM Resin (1 mmol) and add to the reactor, then add DMF (20 mL) to the reactor and nitrogen bubble to swell for 0.5 hours, and then dry.

[0297] Add 20% piperidine / DMF (20 mL) to the reactor and nitrogen bubble for 5 minutes, and then dry. Add 20% piperidine / DMF (20 mL) to the reactor and nitrogen bubble for 20 minutes, and then dry. Wash with DMF (20 mL) for 4 times, 2 minutes each time, and then dry. Ninhydrin test, resin blue.

[0298] Weigh Fmoc-Ser(tBu)-OH (3.0 eq), HOBt (3.0 eq), DIC (3.0 eq) and add to DMF (20 mL), mix well and then add to the reactor, and then nitrogen bubble to react for 2 hours. Ninhydrin test, resin unchanged blue. Wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, DMF in turn.

[0299] 3.2.2.2 Coupling of peptide chain sequence

[0300] According to the sequence of the peptide chain of the compound, the sequence of the peptide chain is N-terminal to C-terminal (H-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Leu-Leu-Glu-Lys-Lys-Ala-Ala-Lys-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2). The solid-phase synthesis is synthesized in the order of C-terminal to N-terminal. The amount of the protected amino acid and the condensation reagent and its condensation method are the same as those of the coupling of Fmoc-Ser(tBu)-OH to Rink Amide-AM Resin. The protected amino acids used in the synthesis process are Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc Gly-OH, Fmoc-Ala-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Lys(ivDde)-OH, Fmoc-Ala-OH, Fmoc-Ala-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Ser(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-His(Trt)-Gly-OH, Boc-His(Trt)-Aib-OH, respectively. The coupling of the protected amino acid and the deprotection of Fmoc are repeated to obtain a linear resin peptide.

[0301] 3.2.3.3. Removal of ivDde protecting group from linear resin peptide

[0302] To the reactor containing the linear resin peptide, add 3% hydrazine hydrate / DMF (20 mL), and bubble nitrogen for 20 minutes, and then drain. Repeat this step once. Wash with DMF (20 mL) for 4 times, 2 minutes each time, and then drain. Ninhydrin test, resin is blue.

[0303] 3.2.2.4 Coupling of Fmoc-Ida(OAll)-OH

[0304] Weigh Fmoc-Ida(OAll)-OH (3.0 eq), HCTU (3.0 eq), and DIEA (6.0 eq) into DMF (20 mL), and then add to the reactor. After mixing, bubble nitrogen for 2 hours. Ninhydrin test, resin is not blue. Wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF, respectively.

[0305] 3.2.3.5 Removal of Fmoc protecting group

[0306] To the reactor, add 20% piperidine / DMF (20 mL), and bubble nitrogen for 5 minutes, and then drain. To the reactor, add 20% piperidine / DMF (20 mL), and bubble nitrogen for 20 minutes, and then drain. Wash with DMF (20 mL) for 4 times, 2 minutes each time, and then drain. Ninhydrin test, resin is blue.

[0307] 3.2.3.6 Side chain coupling

[0308] Weigh the side chain molecule (3.0 eq, CAS: 1188328-37-1), HCTU (3.0 eq), and DIEA (6.0 eq) into DMF (20 mL), and then add to the reactor. After mixing, bubble nitrogen for 4 hours. Ninhydrin test, resin is not blue. Wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF, respectively.

[0309] To the reactor, add DCM (20 mL), and bubble nitrogen for 2 minutes, and then drain. Repeat this step once. Weigh morpholine (10 eq) and Pd(PPh3)4 (0.2 eq) into DCM (20 mL), and then add to the reactor. Bubble nitrogen for 20 minutes, and then drain. Repeat this step twice. Wash with DCM (20 mL) for 3 times, 2 minutes each time, and then drain. Wash with DMF (20 mL) for 3 times, 2 minutes each time, and then drain. Ninhydrin test, resin is blue.

[0310] 3.2.3.8 Linear resin peptide amidation ring

[0311] Weigh HCTU (3.0 eq), DIEA (6.0 eq) into DMF (20 mL), mix well, then add into the reactor, nitrogen bubbling reaction for 4 hours. Indanetrione detection, resin does not turn blue. Wash with DMF, isopropyl alcohol, DMF, isopropyl alcohol, DMF, DMF, DMF in turn.

[0312] 3.2.3.9 Cleavage of resin peptide

[0313] Prepare 50 mL cleavage solution according to the ratio of TFA: DTT: Tis: H2O = 92.5: 2.5: 2.5: 2.5. Add the resin peptide into the cleavage solution, stir at room temperature for 2 hours. After the reaction is completed, filter, and elute the filtrate into isopropyl ether (500 mL), filter to obtain the crude peptide.

[0314] 3.2.3.10 Purification of crude peptide

[0315] Dissolve the crude peptide in 20% acetonitrile / water solution, filter through a 0.45 um membrane, and then separate by a reverse phase high performance liquid chromatography system, with buffer A (0.1% trifluoroacetic acid aqueous solution) and B (acetonitrile). Among them, the chromatographic column is a C18 reverse phase chromatographic column, and the chromatographic detector wavelength is set to 220 nm during purification, and the flow rate is 70 mL / min. Collect the target components, concentrate the reverse phase column, and then concentrate and freeze-dry to obtain the product. The sample is determined by HPLC to have a purity of 90%; LC / MS (ESI): [(M-4H) / 4] - =1236.72, which is consistent with the molecular weight of the compound 4951.7.

[0316] 3.2.4 Chemical synthesis of compound No. 29 cyclo(17,21)H-Aib-H-G-T-F-T-S-D-Y-S-I-L-L-E-K-K-A-A-Q-K(-COCH2N(-OEG-OEG-γGlu-C20diacid)- CH2CO)-F-V-E-W-L-L-A-G-G-P-S-S-G-A-P-P-P-S-NH2

[0317] 3.2.4.1 Coupling of Fmoc-Ser(tBu)-OH with Rink Amide-AM Resin

[0318] Weigh Rink Amide-AM Resin (1 mmol) into the reactor, then add DMF (20 mL) into the reactor, nitrogen bubbling swelling for 0.5 hours, and dry the solvent.

[0319] To the reactor containing the linear resin peptide, 20% piperidine / DMF (20 mL) was added and nitrogen was bubbled for 5 minutes. The solvent was removed by suction. To the reactor, 20% piperidine / DMF (20 mL) was added and nitrogen was bubbled for 20 minutes. The solvent was removed by suction. DMF (20 mL) was added and washed for 2 minutes. The solvent was removed by suction. This was repeated 3 times. The resin was checked by ninhydrin test. The resin was blue.

[0320] Fmoc-Ser(tBu)-OH (3.0 eq), HOBt (3.0 eq), DIC (3.0 eq) were weighed into DMF (20 mL) and added to the reactor. After mixing, nitrogen was bubbled for 3 hours. The resin was checked by ninhydrin test. The resin was not blue. The reaction was removed by suction and washed with DMF, isopropanol, DMF, isopropanol, DMF, DMF, DMF, respectively.

[0321] 3.2.4.2 Coupling of the peptide chain sequence

[0322] According to the sequence of the peptide chain of the compound, the sequence of the peptide chain is N-terminal to C-terminal (H-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Leu-Leu-Glu-Lys-Lys-Ala-Ala-Gln-Lys-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2). The solid-phase synthesis is synthesized in the order of C-terminal to N-terminal. The amount of the protected amino acid and the condensation reagent and the condensation method thereof are the same as those of the coupling of Fmoc-Ser(tBu)-OH to Rink Amide-AM Resin. The protected amino acids used in the synthesis process are Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Lys(ivDde)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH, Fmoc-Ala-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Ser(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-His(Trt)-Gly-OH, Boc-His(Trt)-Aib-OH, respectively. The coupling of the protected amino acid and the deprotection of Fmoc are repeated to obtain a linear resin peptide.

[0323] 3.2.4.3 Side chain coupling and lactam ring

[0324] 3.2.4.3.1 Removal of the Alloc protecting group from the linear resin peptide

[0325] To the reactor, add DCM (20 mL), and bubble nitrogen for 5 minutes. Dry the solvent. Repeat this step once. Weigh out morpholine (10 eq) and Pd(PPh3)4(0.2 eq) into DCM (20 mL), and add to the reactor. Bubble nitrogen for 20 minutes. Dry the solvent. Repeat this step twice. Wash with DCM (20 mL) for 3 times, 2 minutes each. Dry the solvent. Wash with DMF (20 mL) for 3 times, 2 minutes each. Dry the solvent. Ninhydrin test, resin blue.

[0326] 3.2.4.3.2 Coupling of Fmoc-Ida(OAll)-OH

[0327] Weigh out Fmoc-Ida(OAll)-OH (3.0 eq), HCTU (3.0 eq), and DIEA (6.0 eq) into DMF (20 mL), and add to the reactor. Bubble nitrogen for 24 hours. Ninhydrin test, resin unchanged. Dry the solvent, and wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF.

[0328] 3.2.4.3.3 Removal of Fmoc protecting group

[0329] To the reactor, add 20% piperidine / DMF (20 mL), and bubble nitrogen for 5 minutes. Dry the solvent. To the reactor, add 20% piperidine / DMF (20 mL), and bubble nitrogen for 20 minutes. Dry the solvent. Wash with DMF (20 mL) for 4 times, 2 minutes each. Dry the solvent. Ninhydrin test, resin blue.

[0330] 3.2.4.3.4 Side chain coupling

[0331] Weigh out side chain molecule (3.0 eq, CAS: 1188328-37-1), HCTU (3.0 eq), and DIEA (6.0 eq) into DMF (20 mL), and add to the reactor. Bubble nitrogen for 24 hours. Ninhydrin test, resin unchanged. Dry the solvent, and wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF.

[0332] 3.2.4.3.5 Removal of OAll protecting group

[0333] To the reactor, add DCM (20 mL), and bubble nitrogen for 5 minutes. Repeat this step once. Weigh morpholine (10 eq) and Pd(PPh3)4(0.2 eq) into DCM (20 mL), and add to the reactor. Bubble nitrogen for 20 minutes. Repeat this step twice. Wash with DCM (20 mL) for 3 times, 2 minutes each time. Dry the solvent. Wash with DMF (20 mL) for 3 times, 2 minutes each time. Dry the solvent.

[0334] 3.2.4.3.6 Removal of ivDde protecting group from linear resin peptide

[0335] To the reactor, add 3% hydrazine hydrate / DMF (20 mL), and bubble nitrogen for 10 minutes. Repeat this step twice. Wash with DMF (20 mL) for 4 times, 2 minutes each time. Dry the solvent. Ninhydrin test, resin is blue.

[0336] 3.2.4.3.7 Closure of the amide ring

[0337] Weigh HATU (3.0 eq), HOBt (3.0 eq), and DIEA (6.0 eq) into DMF (20 mL), and add to the reactor. Bubble nitrogen for 4 hours. Ninhydrin test, resin does not turn blue. Dry the reaction, and wash with DMF, isopropanol, DMF, isopropanol, DMF, DMF, and DMF, in sequence.

[0338] 3.2.4.4 Cleavage of the resin peptide

[0339] Prepare 40 mL cleavage solution according to the ratio of TFA: DTT: Tis: H2O = 92.5: 2.5: 2.5: 2.5. Add the resin peptide to the cleavage solution, and stir at room temperature for 2 hours. After the reaction is completed, filter, and elute the filtrate into isopropyl ether (400 mL). Filter, and obtain the crude peptide.

[0340] 3.2.4.5 Purification of the crude peptide

[0341] Dissolve the crude peptide in 20% acetonitrile / water solution, filter through a 0.45 um membrane, and separate using a reverse-phase high-performance liquid chromatography system. The purification mobile phase is A (0.1% trifluoroacetic acid aqueous solution) and B (acetonitrile). The chromatographic column is a C18 reverse-phase chromatographic column, the chromatograph detection wavelength is set to 220 nm during the purification process, and the flow rate is 70 mL / min. Collect the target component, concentrate after reverse-phase column, and freeze-dry to obtain the product. The sample is determined for purity by HPLC, 90.0%; LC / MS (ESI): [(M-4H) / 4] = 1236.83, which is consistent with the molecular weight of the compound 4950.7. -

[0342] ​Compounds No. 30-51 and 61-64 of the present application can be synthesized according to the test protocols of the above examples.

[0343] Some of the compounds of the present application were partially synthesized and their purity and structure were confirmed by analytical UPLC and LC / MS as shown in the following table:

[0344] Biological test evaluation

[0345] The following test examples further illustrate the present application but are not meant to limit the scope of the present application.

[0346] 1. Experimental instruments

[0347] Microplate reader (BioTek Synergy H1), Pipettor (Eppendorf & Rainin), IDOT non-contact microdispensing system (Dispendix)

[0348] 2. Experimental reagents:

[0349] DMEM / F12 medium was purchased from Gibco, item No. 11330032, casein was purchased from Sigma, item No. C4765-10ML, 384-well plate was purchased from PerkinElmer, item No. 6007299, IBMX was purchased from Sigma, item No. I7018, CisbiocAMP-GsDynamickit was purchased from Cisbio, item No. 62AM4PEC, 96-well plate was purchased from Corning, item No. 3610

[0350] Test Example 1: Determination of the ability of the compounds of the present application to stimulate the production of cAMP by human GLP1R, GIPR, GCGR stable cell lines respectively under 0.1% casein condition

[0351] 1. Experimental purpose: The purpose of this test example is to test the ability of the compounds to generate cAMP after activating human GLP-1R, GIPR, GCGR on the cell surface.

[0352] 2. Experimental method

[0353] CHO-K1 / hGLP-1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR stable cell lines were trypsinized and centrifuged, resuspended with DMEM / F12 (Gibco Cat#11330032) complete medium, counted, and plated at 7500 cells / 100 μL / well in 96-well cell plates, and incubated at 37°C, 5% CO2 incubator overnight. The next day, the plates were taken out, the supernatant was discarded, and the cells were washed once with serum-free DMEM / F12, and then 100 μL / well of DMEM / F12 medium containing 500 μM IBMX (Sigma Cat#I7018) and 0.1% casein (Sigma Cat#C4765) was added. 1 μL / well of polypeptide sample (1 nM starting, 3-fold dilution, 12 concentrations) diluted with DMSO was added using an IDOT dispenser, and incubated at 37°C for 30 minutes. Detection was performed using the cAMP detection kit Cisbio cAMP-Gs Dynamic kit (Cisbio Cat#62AM4PEC), the plates were taken out, the supernatant was discarded, and the cells were washed once with PBS, and 50 μL / well of cAMP Lysis & Detection Buffer in the kit was added as a lysis solution, and the cells were shaken and lysed at room temperature for 5 minutes. 10 μL / well of cell supernatant lysate was aspirated and added to a 384-well plate, 5 μL / well of cAMP-d2 solution diluted 20 times with the lysis solution was added, and 5 μL / well of Anti-cAMP-Eu 3+ -Cryptate solution, shake for 10 seconds, mix well, and incubate at room temperature for 1 hour in the dark.

[0354] 3. Experimental data processing method:

[0355] 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 Graph Pad Prism 9 to obtain the EC 50 value.

[0356] 4. Experimental conclusion: Through the above scheme, it is concluded that the compound of the present application shows good biological activity in the CHO-K1 / hGLP1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR cell agonist activity test after stimulating the CHO-K1 / hGLP1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR stable cell line for 30 minutes under the condition of 0.1% casein.

[0357] Test Example Two: Assay for the ability of the compounds of the present application to stimulate cAMP production in CHO-K1 / hGLP-1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR stable cell lines in the presence of 100% human plasma

[0358] 1. Purpose of the experiment: The purpose of this test example is to test the ability of the compounds to activate the production of cAMP in stable cell lines expressing human GLP-1R, GIPR, GCGR on the cell surface.

[0359] 2. Experimental method: CHO-K1 / hGLP-1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR stable cell lines were trypsinized and centrifuged, resuspended with DMEM / F12 (Gibco Cat#11330032) complete medium, counted, and plated at 7500 cells / 100 μL / well in 96-well cell plates and incubated overnight at 37°C in a 5% CO2 incubator. The next day, the cell plates were removed, the supernatant was discarded, and the cells were washed once with serum-free DMEM / F12 and then 100% human plasma solution containing 500 μM IBMX (Sigma Cat#I7018) was added at 100 μL per well. 1 μL / well of polypeptide samples diluted in DMSO (10 μM or 1 μM starting, 3-fold dilution, 12 concentrations) was added using an IDOT dispenser and incubated at 37°C for 30 minutes. Detection was performed using the cAMP assay kit Cisbio cAMP-Gs Dynamik kit (Cisbio Cat#62AM4PEC), the cell plates were removed, the supernatant was discarded, and the cells were washed once with PBS and then cAMP Lysis & Detection Buffer from the kit was added as lysis solution at 50 μL per well and incubated at room temperature for 5 minutes with shaking. 10 μL of cell supernatant lysate was removed per well and added to a 384-well plate, 5 μL / well of cAMP-d2 solution diluted 20-fold in lysis solution was added, and 5 μL / well of Anti-cAMP-Eu3+-Cryptate solution diluted 20-fold in lysis solution was added, the mixture was shaken for 10 seconds, and incubated at room temperature for 1 hour in the dark.

[0360] 3. Experimental data processing method: 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 Graph Pad Prism 9 to obtain the EC50 value.

[0361] 4. Experimental conclusion: Through the above scheme, it is concluded that the compound of the present application shows good agonistic activity in the CHO-K1 / hGLP1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR cell agonistic activity test after stimulating CHO-K1 / hGLP1R, CHO-K1 / hGIPR, CHO-K1 / hGCGR stable cell strains for 30 minutes in 100% human plasma conditions to produce cAMP.

[0362] Test Example Three: Determination of the affinity of the compound of the present application to human GLP1 receptor

[0363] Purpose of the experiment: The purpose of this test example is to test the affinity of the compound to human GLP-1 receptor by using isotope competitive binding method.

[0364] Experimental reagents and instruments

[0365] 2.1 Experimental instruments:

[0366] Mico Beta2 Reader (Perkin Elmer, Model CNLL0153)

[0367] Plate washing machine (Perkin Elmer, Model UNIFILTER-96)

[0368] 2.2 Experimental reagents and materials

[0369] [125I]-GLP-1 was purchased from Perkin Elmer, Catalog No. NEX308

[0370] GLP-1 was purchased from HaoYuan, Catalog No. HY-P0055

[0371] HEPES was purchased from Gibco, Catalog No. 15630-080

[0372] EGTA was purchased from Aladdin, Catalog No. E104432-500g

[0373] BSA was purchased from Sigma, Catalog No. B2064-100G

[0374] HSA was purchased from Sigma, Catalog No. A3782

[0375] MgCl2 was purchased from Sigma, Catalog No. M1028

[0376] Tween20 was purchased from Energy, Catalog No. E080756

[0377] Microscint Ococktail was purchased from Perkin Elmer, Catalog No. 6013611

[0378] Sealing membrane was purchased from Perkin Elmer, item number 6050185

[0379] 96 GF / C filter plate was purchased from Perkin Elmer, item number 6055690

[0380] 96-well polypropylene plate was purchased from Agilent, item number 5043-9311

[0381] Experimental method:

[0382] Human GLP-1 receptor membrane protein was extracted from a stable cell line overexpressing GLP-1 receptor by WuXi Biotechnology (Shanghai) Co., Ltd. The collected cell mass of the stable cell line overexpressing GLP-1 receptor was dissolved with 50 mM Tris-HCl, and the rotor was stirred in an ice box for 12 times or more. The supernatant was obtained by low-speed centrifugation, and the precipitate was obtained by high-speed centrifugation. Finally, the precipitate was aliquoted with pre-cooled dissolving solution (50 mM Tris-HCl, 10 mM MgCl2, 0.5 mM EDTA, 10% glycerol) to obtain GLP-1 receptor cell membrane. The experiment was divided into two conditions according to the different concentrations of HSA (human serum albumin): 0% HSA and 2% HSA, in order to evaluate the effect of serum protein binding on the affinity of the compound.

[0383] Under the condition of 0% HSA, the test compound was diluted with DMSO to the corresponding working concentration (8 concentration points, four-fold dilution, double duplicate wells) and added to 1 μL in a 96-well polypropylene plate. The negative control added 1 μL of GLP-1 solution with a final concentration of 500 nM, the positive control added 1 μL of detection buffer, then 50 μL of 0% HSA detection buffer (50 mM HEPES, 5 mM EGTA, 5 mM MgCl2, 0.005% Tween-20, pH 7.4) diluted GLP-1 receptor cell membrane and 50 μL of 0% HSA detection buffer diluted [125I]-GLP-1 were added to the 96-well polypropylene plate. At this time, the final concentration of the compound under the condition of 0% HSA was 1000 nM-0.06 nM, and the final concentration of GLP-1 was 500 nM-0.03 nM.

[0384] The test compound is diluted with DMSO to the corresponding working concentration (8 concentration points, four-fold dilution, double duplicate wells) under 2% HSA condition and 1 μL is added to a 96-well polypropylene plate, 1 μL of GLP-1 solution with a final concentration of 500 nM is added to the negative control, 1 μL of detection buffer is added to the positive control, and then 50 μL of GLP-1 receptor cell membrane diluted with 2% HSA detection buffer (50 mM HEPES, 5 mM EGTA, 5 mM MgCl2, 0.005% Tween-20, 2% HSA, pH 7.4) and 50 μL of [125I]-GLP-1 diluted with 2% HSA detection buffer are added to the 96-well polypropylene plate, at this time the final concentration of the compound under 2% HSA condition is 20,000 nM-1.2 nM, and the final concentration of GLP-1 is 500 nM-0.03 nM.

[0385] The 96-well polypropylene plate under 0% HSA and 2% HSA conditions is sealed with a sealing film, and then incubated at room temperature on a shaker for 1 hour, while the GF / C filter plate (Unifilter-96 GF / C filter plate) is soaked in the plate soaking buffer (50 mM HEPES, 0.5% BSA, pH 7.4) for at least 0.5 hours. After incubation, the reaction solution is collected on the GF / C filter plate with a cell collector, washed 6 times with the plate washing buffer (50 mM HEPES, 5 mM EGTA, 5 mM MgCl2, 0.005% Tween-20, pH 7.4), and dried in a 50°C oven for 1 hour. The dried GF / C filter plate is sealed at the bottom, 50 μL of scintillation solution is added to each well, and sealed. The Microbeta2 is used for reading.

[0386] Experimental data processing method:

[0387] Inhibition rate % = 100% - (sample well signal value - negative control) / (positive control - negative control) * 100%

[0388] Fitting is performed with "log (antagonist) vs. response - Variable slope" in Graph Pad Prism to calculate the IC50.

[0389] The formula for calculating Ki is: Ki = IC50 / (1+(isotope concentration / Kd)) The Kd value is 1321 pM

[0390] Experimental conclusion:

[0391] Through the above scheme, the competitive affinity of the compound of the present application to the human GLP-1 receptor is shown as follows.

[0392] Test Example Four: Pharmacokinetic evaluation test in cynomolgus monkeys

[0393] 1. Research purposes: To study the pharmacokinetic behavior of the compound of the present application in cynomolgus monkeys (plasma) at a dose of 0.02 mg / kg intravenous administration.

[0394] 2. Test scheme

[0395] 2.1 Test drug: Compound of the present application, self-made.

[0396] 2.2 Test animals: 3 cynomolgus monkeys per group, male, Hainan Xinzhenyuan Biotechnology Co., Ltd., Animal Production License No.: SCXK (Qiong) 2021-0002.

[0397] 2.3 Formulation prescription: Intravenous drug preparation: PBS

[0398] Weigh the compound of the example, add PBS by volume, and ultrasonicate to make it completely dissolved to obtain a colorless transparent clear solution with a concentration of 0.01 mg / mL.

[0399] 2.4 Drug administration: 3 cynomolgus monkeys, male, fasted overnight, respectively I.V.; dose 0.02 mg / kg, drug volume 2 mL / kg.

[0400] 2.5 Sample collection: Before and after drug administration, 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 24, 48, 72, 96, 120, 144, 168 h, peripheral venous puncture blood collection, whole blood in EDTA-K2 test tube, 4℃ 4000xg centrifugation 5min separation plasma, -80℃ preservation.

[0401] 2.6 Sample processing:

[0402] Add 50uL of plasma sample to 200uL of acetonitrile for precipitation, mix and centrifuge at 3200xg for 10 minutes.

[0403] Take 100uL of the supernatant after treatment and mix with 200uL of water, and analyze the concentration of the test compound by LC / MS / MS.

[0404] 2.7 Biological analysis

[0405] Mass spectrometry conditions

[0406] Mass spectrometer ABSciex QTRAP6500+ mass spectrometer;

[0407] Ion source: electrospray ionization source (ESI) dry gas: N2

[0408] Electrospray voltage: 5500V detection mode: positive ion detection

[0409] Scan mode: reaction monitoring (MRM) mode

[0410] Chromatographic conditions

[0411] Liquid phase instrument: Shimadzu LC40 liquid phase system;

[0412] Chromatographic column: XBridge Peptide BEHC18 3.5 μm (2.1 x 100 mm);

[0413] Mobile phase: A phase is 0.1% formic acid in 95% water solution, B liquid is 0.1% formic acid in 95% acetonitrile solution;

[0414] Flow rate: 0.50 mL / min; running time: 3 min; sample volume: 10 μL;

[0415] Elution gradient:

[0416] 3. Test results and analysis: The main pharmacokinetic parameters were calculated by Win Nonlin 8.2, and the pharmacokinetic test results of cynomolgus monkeys are shown in the following table:

[0417] 4. Experimental conclusion: The data show that the compound of the preferred embodiment of the application has high exposure, long half-life and low clearance in the cynomolgus monkey pharmacokinetic evaluation experiment.

[0418] Test example five: SD rat pharmacokinetic determination

[0419] 1. Purpose of the study:

[0420] SD rats were used as test animals to study the pharmacokinetic behavior of the compound of the application in rat plasma after intravenous injection at a dose of 1 mg / kg.

[0421] 2. Test scheme

[0422] 2.1 Test drug:

[0423] The compound of the embodiment of the application was self-made.

[0424] 2.2 Test animals:

[0425] SD rats 3 per example, male, Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd., animal production license number: SCXK (Zhe) 2024-0001).

[0426] 2.3 Drug preparation:

[0427] Intravenous drug preparation: PBS

[0428] The example compound was weighed out and added to PBS by volume to give a clear colourless solution with a concentration of 0.2 mg / mL.

[0429] 2.4 Administration:

[0430] SD rats, male; after fasting overnight, administered i.v. at a dose of 1 mg / kg, with a dose volume of 5 mL / kg.

[0431] 2.5 Sample collection:

[0432] Blood samples (0.2 mL) were taken from the jugular vein of the rats before administration and at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 24 and 48 h after administration, and placed in EDTA-K2 tubes. The plasma was separated by centrifugation at 4000 x g for 5 min at 4°C and stored at -80°C.

[0433] 2.6 Sample processing:

[0434] 55 μL of the plasma sample was added to 3 μL of 5% aqueous trichloroacetic acid (TCA) and vortexed, then 200 μL of acetonitrile / methanol solution containing an internal standard (v:v = 1:1) was added and vortexed to precipitate the proteins. After centrifugation at 4000 rpm for 15 min at 4°C, the supernatant was diluted 3-fold with purified water.

[0435] The diluted supernatant was injected into the LC / MS / MS system for quantitative analysis.

[0436] 2.7 Liquid chromatography analysis

[0437] Liquid chromatography system: Shimadzu LC-40DXS pump

[0438] Mass spectrometry system: AB Sciex API mass spectrometer

[0439] Chromatography column type: Raptor Biphenyl

[0440] Mobile phase: A liquid was 0.1% aqueous formic acid and B liquid was 0.1% formic acid acetonitrile

[0441] Flow rate: 0.6 mL / min

[0442] Elution gradient:

[0443] 3. Test results and analysis

[0444] The main pharmacokinetic parameters were calculated using Win Nonlin 8.2. The results of the rat pharmacokinetic experiment are shown in the table below:

[0445] 4. Experimental Conclusion:

[0446] The data shows that the compound of the embodiment of the application has the remarkable effects of high exposure, long half-life and low clearance rate in the rat pharmacokinetic evaluation experiment.

[0447] Test Example Six: Stability of the compound of the application in blood plasma

[0448] 1. Purpose of the study: to study the stability of the compound of the embodiment in mouse, rat, beagle dog, cynomolgus monkey and human blood plasma.

[0449] 2. Experimental reagents

[0450] The compound of the application (self-made), methanol (Merck), acetonitrile (Merck), DMSO (Sigma), Propantheline (Sigma Lot 1571001), Mevinolin (Sigma Lot 1370600).

[0451] 3. Plasma samples

[0452] 4. Experimental steps

[0453] 1) Solution preparation

[0454] Prepare a 1 mM working solution of the test compound in pure water, a 1 mM working solution of the control compound Melvinolin in DMSO, and a 1 mM working solution of the control compound Propantheline in acetonitrile. Propantheline is used as a positive control for mouse, dog, monkey and human blood plasma, and Melvinolin is used as a positive control for rat blood plasma.

[0455] 2) Stability determination

[0456] Add 398 μL of plasma to each well of the incubation plate and preheat at 37°C for 15 minutes.

[0457] After pre-incubation, add 2 μL of the working solution to 398 μL of plasma and mix well, and then incubate the reaction sample at 37°C. The final incubation concentration of the test compound is 5 μM.

[0458] At 0, 1, 2, 4, 6 and 24 hours, respectively, take 50 μL from the reaction sample, add 450 μL of cold methanol containing formic acid (0.1%) and an internal standard to terminate the reaction.

[0459] Vortex all samples for 10 minutes, and then centrifuge at 3220 g for 30 minutes to precipitate the protein. Transfer 100 μL of the supernatant to a new well plate. According to the LC-MS signal response and peak shape, dilute the supernatant with ultrapure water, and then inject for detection.

[0460] 5. Bioanalysis

[0461] 1) Chromatographic conditions

[0462] Liquid chromatography system: Shimadzu LC-40DXS

[0463] Chromatographic column: 3 pm Horizon C18 50*2.1 mm

[0464] Mobile phase: A phase: 0.1% formic acid in water; B phase: 0.1% formic acid in acetonitrile

[0465] Elution gradient:

[0466] 2) Mass spectrometry conditions

[0467] Mass spectrometer: AB Sciex Triple Quad 6500+

[0468] Ion source: Electrospray ionization source (ESI)

[0469] Dry gas: N2, temperature 500 °C

[0470] Electrospray voltage: 5500 V

[0471] Detection mode: Positive ion detection

[0472] Scan mode: Reaction monitoring (MRM) mode

[0473] 6. Experimental results and data processing

[0474] Peak area ratio = compound peak area / internal standard peak area

[0475] Percentage of compound remaining (t hours) = peak area ratio t hours / peak area ratio 0 hours x 100

[0476] 7. Experimental conclusion:

[0477] The preferred embodiment compounds of the present application all exhibit excellent stability in the plasma of various genera. Test Example Seven: Investigation of the in vitro metabolic stability of the compounds of the present application in liver microsomes

[0478] 1. Purpose of the study: To study the phase I metabolic stability of the compounds of the present example in mouse, rat, beagle dog, cynomolgus monkey and human liver microsomes.

[0479] 2. Experimental reagents

[0480] Compounds (self-made), PBS (Gibco pH 7.4 Lot 2842949), NADPH (Sigma Lot 481973), Methanol (Merck), Acetonitrile (Merck), DMSO (Sigma), Verapamil (Sigma Lot 1711202).

[0481] 3. Liver microsomes

[0482] 4. Drug configuration

[0483] 1) Compound stock solution preparation: The test compound was configured into 10 mM stock solution with DMSO, and stored in the refrigerator at -20°C for later use.

[0484] 2) Compound working solution preparation: 10 μL of the compound stock solution was taken, and 990 μL of PBS was added to configure the compound working solution into 100 μM. According to the properties of the compound, the configuration ratio can be appropriately adjusted.

[0485] 3) NADPH configuration: NADPH was dissolved into 10 mM working solution with PBS.

[0486] 4) Preparation of reaction termination solution: The internal standard was diluted with acetonitrile / water (v:v = 1:1) to prepare the termination solution (200 nM labetalol, 100 nM ketoprofen, 100 nM tolbutamide), and stored in the refrigerator at 2-8°C.

[0487] 5. Incubation process

[0488] 1) In the 96-well plate, 216.25 μL of PBS, 6.25 μL of 20 mg / mL liver microsomes, and 25 μL of 10 mM NADPH were sequentially added, and pre-incubated at 37°C for 10 min.

[0489] 2) 2.5 μL of 100 μM compound working solution was added to start the reaction, and incubated at 37°C. The total volume of the reaction system was 250 μL, and the final content of each component was: the compound of the present embodiment (1 μM), liver microsomes (0.5 mg / mL), and NADPH (1 mM).

[0490] 3) At 0, 5, 15, 30, and 60 min time points, 30 μL was taken out, 150 μL of cold termination solution containing the internal standard was added to terminate the reaction, 3220 g centrifugation was performed for 40 min, 100 μL of supernatant was mixed with 100 μL of pure water, and LC-MS / MS analysis was performed.

[0491] 6. Biological analysis

[0492] 1) Chromatographic conditions

[0493] Liquid phase system: Shimadzu

[0494] Mobile phase: Phase A: 0.1% formic acid in water; Phase B: 0.1% formic acid in acetonitrile

[0495] Chromatographic column: HSST32.5μm2.1×50mmColumnXP

[0496] Elution gradient:

[0497] 2) Mass spectrometry conditions

[0498] Instrument: Triple Quad TM 6500+ABSciex Triple Quad5500+

[0499] Ion source: Electrospray ionization source (ESI)

[0500] Dry gas: N2

[0501] Electrospray voltage: 5500V

[0502] Detection mode: Positive ion detection

[0503] Scan mode: Reaction monitoring (MRM) mode

[0504] 7. Data processing

[0505] The raw data is calculated according to the following formula:

[0506] Remaining rate % = peak area ratio of compound to internal standard at any time point / peak area ratio of compound to internal standard at 0 minute x 100

[0507] 8. Experimental conclusion:

[0508] The results show that the compound of the preferred embodiment of the present application presents a stable metabolic effect in liver microsomes of various genera.

[0509] Test Example Eight: Effect of long-term administration of the compound of the present application on body weight and food intake of DIO model C57 mice fed with high-fat feed

[0510] 1. Experimental purpose: The purpose of this test is to evaluate the effect of long-term administration of the compound on the body weight and food intake of DIO model C57BL / 6 mice fed with high-fat feed.

[0511] 2. Experimental reagents and instruments

[0512] C57BL / 6, male, 16-17 weeks, purchased from Zhejiang Vintone Lihua Experimental Animal Technology Co., Ltd.

[0513] 60% high-fat diet (HFD) purchased from Shanghai Biopsy Company (D12492, Research Diet)

[0514] Electronic balance (BSA2202S-CW, Sartorius)

[0515] 3. Experimental method

[0516] 3.1 140 C57BL / 6 mice induced by high-fat diet for 10 weeks and 10 normal control mice were purchased from Weitonglihua (Beijing) Pharmaceutical Technology Co., Ltd. After the animals arrived, the high-fat diet induction group continued to be fed with high-fat diet for 1-2 weeks, and the normal control diet group continued to be fed with normal diet. After the body weight was stable, the experiment began.

[0517] 3.2 After 1-2 weeks of adaptive feeding, the modeling group animals were randomly divided into 12 groups according to body weight, 10 in each group. The first group was the Vehicle group (Vehicle: SSC buffer), which was given the solvent; the remaining groups were the drug administration groups; the drug administration scheme: subcutaneous injection of the corresponding compound, the administration cycle was 21 days, once every three days, and the administration volume was 5 mL / kg. The Blank group continued to be fed with normal diet, and the solvent was subcutaneously injected once every three days.

[0518] 3.3 The first day of administration was defined as Day 0, and the body weight of each group of mice was weighed and recorded every morning. The food intake of each group of mice was measured once every three days, and the specific method was to replace the feed after each weighing and administration, and record the added and remaining amounts.

[0519] 3.4 Each time the animals were weighed and the data were recorded, and the subcutaneous injection was given according to the body weight, and the administration volume was 5 mL / kg.

[0520] 3.5 On Day 16, 50 microliters of blood were taken from each group of mice at 0.5h, 1h, 2h (N=3), 4h, 6h, 8h (N=3), 24h, 48h, 72h (N=4) after administration, and the plasma was obtained by centrifugation.

[0521] 3.6 On Day 18 at 17:00, all animals were fasted overnight without water.

[0522] 3.7 On day 19, blood samples were taken from the tail tip of each animal in the morning to measure the fasting blood glucose level. Then, about 50 μL of blood was taken, centrifuged to obtain the plasma, which was used for PK detection. The animals were packed and taken to measure the body fat rate, the lean mass and fat mass, and the lean mass / fat mass ratio. All mice were euthanized in order according to the grouping sequence, and at least 500 uL of blood was taken from the heart, from which 120 uL of plasma was absorbed for blood biochemical detection (ALT, AST, TG, TC, LDL, HDL), and 100 uL of plasma was tested for insulin level. After dissection, the whole liver was taken and weighed to calculate the liver / body weight ratio, and part of the liver sample was quickly frozen with dry ice for TG testing.

[0523] 4. Experimental data processing and statistical analysis

[0524] The body weight and body weight change rate of the mice after administration were summarized and statistically analyzed. The body weight change rate was calculated as (BWt-BW0) / BW0x100%. BWt represents the body weight of the mice on the tth day of the experiment, and BW0 represents the body weight of the mice on the 0th day of the experiment.

[0525] The food intake was calculated as (added amount (g)-remaining amount (g)) / number of animals per cage, and the cumulative food intake was the total of the food intake of each animal per day during the administration period.

[0526] The experimental data were analyzed and plotted using GraphPad Prism software. The t-test method was used for comparison between two groups. The one-way ANOVA method was used for comparison among three or more groups. p<0.05 was defined as statistically significant difference.

[0527] 5. Experimental conclusion:

[0528] The experimental results show that the compound of the preferred embodiment of the present application can significantly reduce the body weight of DIO obese mice, showing a good dose dependence (-13% to -49%), and the body weight reduction rate at the same dose is greater than that of the control LY3437943. At the same time, it can also significantly reduce the body fat rate, which can be up to more than 20%.

Claims

1. A polypeptide comprising a compound of the general formula (I) or a pharmaceutically acceptable salt thereof, R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -Leu-Glu-X 16 -X 17 -Ala- X 19 -X 20 -X 21 -Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2 (I) wherein, R1is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acetyl, substituted or unsubstituted formyl or substituted or unsubstituted benzoyl; R2is selected from -NH2or -OH; X 10 an amino acid residue selected from Tyr, Fae, modified or unmodified Lys or Y1; X 12 an amino acid residue selected from lie or a modified or unmodified Lys; X 13 an amino acid residue selected from Leu, a-Me-Leu, a-Et-Leu, a-Et-Tyr, Achx, a-Me-cpAla, 4diFAchx, THP, THT, THS, or modified or unmodified Lys; X 16 an amino acid residue selected from a modified or unmodified Lys or Y1; X 17 an amino acid residue selected from modified or unmodified Lys; X 19 an amino acid residue selected from Ala or modified or unmodified Lys; X 20 an amino acid residue selected from Gin or a modified or unmodified Lys; X 21 an amino acid residue selected from Glu or a modified or unmodified Lys; Y1is a Fae or Lys residue to which the substituents of Y2are attached; Or, any two adjacent or non-adjacent amino acid residues can optionally be further linked to form a modified or unmodified ring.

2. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to claim 1, characterized in that, Formula (I) is further illustrated by Formula (II): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-Ala- Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(II) wherein: X 13 an amino acid residue selected from the group consisting of α-Et-Leu, α-Et-Tyr, Achx, α-Me-cpAla, 4diFAchx, THP, THT, or THS; X 16 Y1is as defined above.

3. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to claim 1, characterized in that, Formula (I) is further represented by Formula (IV): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X 12 -X 13 -Leu-Glu-X 16 -Lys-Ala- Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(IV).

4. The polypeptide according to claim 1, wherein, 5 Formula (I) is further represented by Formula (V): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-Ala-Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(V).

5. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to claim 1, characterized in that, Formula (I) is further illustrated by Formula (VII): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-X 19 - Gln-Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(VII).

6. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to claim 1, characterized in that, Formula (I) is further shown in Formula (VIII): R1-His-Aib-His-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-X 13 -Leu-Glu-X 16 -Lys-Ala-Ala- X 20 -Glu-Phe-Val-Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R2(VIII).

7. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to any one of claims 1-6, characterized in that, the two adjacent or non-adjacent modified amino acid residues are selected from X 16 or X 17 and its adjacent or non-adjacent amino acid residues; preferably X 12 and X 16 , X 13 and X 16 , X 13 and X 17 , X 16 and X 19 , X 16 and X 20 , X 17 and X 20 or X 17 and X 21 ; Further, the any two adjacent or non-adjacent amino acid residues are preferably the amino acid residues of Lys.

8. The polypeptide according to any one of claims 1 to 7, characterized in that, It comprises a polypeptide sequence as shown in any of the following:

9. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to any one of claims 1-8, characterized in that, Y1is a Fae or Lys residue to which the substituents of Y2are attached; Y2 is selected from (NEG) a -(OEG) b -(y-Glu) c -C16 diacid, (NEG) a -(OEG) b -(y-Glu) c -C18 diacid, (NEG) a -(OEG) b -(y-Glu) c -C20 diacid, PEG n -(y-Glu) c -C16, (NEG) a -(OEG) b -(y-Glu) c -C24 diacid, (NEG) a -(OEG) b -(y-Glu) c -CO(CH2) 18 PO(OH)2, (NEG) a -(OEG) b -(y-Glu PA) c -C16 diacid, (NEG) a -(OEG) b -(y-Glu PA) c -C18 diacid, (NEG) a -(OEG) b -(y-Glu PA) c -C20 diacid, PEG n -(y-Glu PA) c -C16, (NEG) a -(OEG) b -(y-Glu PA) c -C24 diacid or (NEG) a -(OEG) b -(y-Glu PA) c -CO(CH2) 18 PO(OH)2; a is 0, 1, 2 or 3; b is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; c is 0, 1, 2 or 3; n is 16, 18, 20, 22 or 24; Preferably, a is 0 or 1, b is 1, 2 or 3, c is 1, and n is 24.

10. The polypeptide and its derivatives, or pharmaceutically acceptable salts thereof, according to any one of claims 1-9, characterized in that, Any two adjacent or non-adjacent amino acid residues can be linked by a modified or unmodified form a cyclic structure; preferably 11. The polypeptide according to any one of claims 1 to 10, wherein the polypeptide is represented by the following formula: ###0001### or a derivative thereof, or a pharmaceutically acceptable salt thereof, wherein X is a halogen atom, and R is a hydrogen atom or a lower alkyl group. R1is selected from hydrogen; R2is selected from -NH2or -OH, preferably -NH2.

12. The polypeptide according to any one of claims 1 to 11, characterized in that, said Y1 is selected from the group consisting of Lys(-NEG-OEG-OEG-γGlu-C16 diacid), Lys(-NEG-OEG-OEG-γGlu-C20 diacid), Lys(-NEG-OEG-γGlu-C20 diacid), Lys(-NEG-OEG-OEG-γGlu-C18 diacid), Lys(-OEG-OEG-γGlu-C18 diacid), Lys(-OEG-OEG-γGlu-C20 diacid), Lys(-PEG 24 -γGlu-C16), Lys(-NEG-OEG-OEG-OEG-γGlu-C20 diacid), Lys(-NEG-OEG-OEG-γGlu-C24 diacid), Lys(-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-OEG-γGluPA-C20 diacid), Lys(-NEG-OEG-OEG-γGluPA-CO(CH2) 18 PO(OH)2), Lys(-NEG-OEG-γGlu-CO(CH2) 18 PO(OH)2) 、 Lys(-NEG-OEG-γGluPA-C20 diacid), Lys(-NEG-OEG-γGluPA-CO(CH2) 18 PO(OH)2) 、 Lys(-NEG-OEG-OEG-γGlu-CO(CH2) 16 PO(OH)2) 、 Lys(-NEG-OEG-OEG-γGluPA–C18 diacid), Lys(-NEG-OEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-γGlu-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2), Lys(-NEG-OEG-yGluPA-C18 diacid), Fae(-NEG-OEG-OEG-yGlu-C16 diacid), Fae(-NEG-OEG-OEG-yGlu-C20 diacid), Fae(-NEG-OEG-yGlu-C20 diacid), Fae(-NEG-OEG-OEG-yGlu-C18 diacid), Fae(-OEG-OEG-yGlu-C18 diacid), Fae(-OEG-OEG-yGlu-C20 diacid), Fae(-PEG 24 -yGlu-C16), Fae(-NEG-OEG-OEG-OEG-yGlu-C20 diacid), Fae(-NEG-OEG-OEG-yGlu-C24 diacid), Fae(-OEG-OEG-yGlu-CO(CH2) 18 PO(OH)2), or Fae(-NEG-OEG-OEG-yGlu-CO(CH2) 18 PO(OH)2), which group has the chemical formula of the following structure:

13. The polypeptide according to any one of claims 1 to 12, characterized in that, said Y2 is selected from the group consisting of -NEG-OEG-OEG-γGlu-C16diacid, -NEG-OEG-OEG-γGlu-C20diacid, -NEG-OEG-γGlu-C20diacid, -NEG-OEG-OEG-γGlu-C18diacid, -OEG-OEG-γGlu-C18diacid, -OEG-OEG-γGlu-C20diacid, -PEG 24 -γGlu-C16, -NEG-OEG-OEG-OEG-γGlu-C20diacid, -NEG-OEG-OEG-γGlu-C24diacid, -OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2, -NEG-OEG-OEG-γGluPA-C20diacid, -NEG-OEG-OEG-γGluPA-CO(CH2) 18 PO(OH)2, -NEG-OEG-OEG-γGlu-CO(CH2) 18 PO(OH)2, -NEG-OEG-γGlu-CO(CH2) 18 PO(OH)2, -NEG-OEG-γGluPA-C20diacid, -NEG-OEG-γGluPA-CO(CH2) 18 PO(OH)2, -NEG-OEG-OEG-γGlu-CO(CH2) 16 PO(OH)2, -NEG-OEG-OEG-γGluPA-C18diacid, -NEG-OEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2, -NEG-OEG-γGlu-CO(CH2) 16 PO(OH)2, -NEG-OEG-γGluPA-CO(CH2) 16 PO(OH)2, or -NEG-OEG-γGluPA-C18diacid has the chemical formula of the following structure:

14. The polypeptide according to any one of claims 1 to 13, characterized in that, Specific structures of the polypeptides and derivatives thereof are as follows:

15. A pharmaceutical composition comprising: 1) a therapeutic amount of the polypeptide according to any one of claims 1-14, derivatives thereof or pharmaceutically acceptable salts thereof, and 2) a pharmaceutically acceptable excipient or pharmaceutical carrier.

16. Use of the polypeptide according to any one of claims 1-14, derivatives thereof or pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 15, in the manufacture of a medicament for activating GLP-1 receptor, GIP receptor and / or GCG receptor; preferably, in the manufacture of a medicament for activating GLP-1, GIP and GCG triple receptor.

17. Use of the polypeptide according to any one of claims 1-14, derivatives thereof or pharmaceutically acceptable salts thereof, and the composition according to claim 15, in the manufacture of a medicament for treating a metabolic disorder-related disease, disorder and / or condition, Preferably, the metabolic disorder-related disease, disorder and / or condition is selected from diabetes or diabetes-related disorder, obesity or obesity-related disorder, non-alcoholic fatty liver.

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