A bis-agonistic polypeptide compound and medical uses thereof

Abstract

This invention discloses a dual-activator polypeptide compound and its pharmaceutical uses, belonging to the field of polypeptide pharmaceuticals. The amino acid sequence of the GIP-R and GLP-1R dual-activator polypeptide compound is: Y-Z-EGTFTSDYSI-X2-LDKI-A-Q-B n -X4-X5-VQWLI-X6-GGPSSG-A-PPPS, with the C-terminal amino acid being amidated. The GIP-R and GLP-1R dual-agonist polypeptide compounds of this invention exhibit more significant dual-agonist activity against human glucose-dependent insulinotropic peptide (GIP) and human glucagon-like peptide-1 (GLP-1) receptors. They not only possess hypoglycemic and weight-loss effects but also have a longer half-life and higher bioavailability, avoiding adverse reactions such as nausea and vomiting caused by long-term use of existing drugs. They can be used as effective raw materials for preparing drugs for treating or preventing diabetes or weight-loss drugs.

Description

Technical Field

[0001] This invention belongs to the field of peptide medicine, specifically relating to a dual-agonist peptide compound and its pharmaceutical uses. This invention is a divisional application with application number 2024109284017, entitled "A Dual-Agonist Peptide Compound and Its Pharmaceutical Uses." Background Technology

[0002] The World Health Organization classifies diabetes into type 1 diabetes, type 2 diabetes, gestational diabetes, and other special types of diabetes, with type 2 diabetes accounting for the vast majority of cases. Type 2 diabetes is primarily related to insulin resistance and pancreatic β-cell defects. Insulin resistance manifests as insulin's inability to function properly in target tissues, leading to metabolic disorders, while pancreatic β-cell defects typically result in insufficient insulin secretion. Because type 2 diabetes often leads to various vascular and non-vascular complications, including obesity, cardiovascular disease, kidney failure, coronary artery disease, microvascular disease, and retinopathy, it currently poses a significant threat to human health. Therefore, the development of safe and effective hypoglycemic drugs is of great significance and value in controlling blood sugar levels in patients with type 2 diabetes.

[0003] Currently, the main medications for treating diabetes include biguanides, sulfonylureas, alpha-glucosidase inhibitors, meglitinides, thiazolidinediones, dipeptidyl peptidase-4 (DPP-4) inhibitors, sodium-glucose cotransporter-2 (SGLT-2) inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, and insulin. Semaglutide, a GLP-1 receptor agonist developed by Novo Nordisk, is one of the most successful drugs for treating diabetes. Compared to other classes of hypoglycemic drugs, semaglutide not only lowers blood sugar but also provides weight loss. In addition, as a GLP-1 receptor agonist, semaglutide also regulates blood lipids and blood pressure, thus lowering blood sugar while reducing the risk of cardiovascular events and providing better safety. Its target, GLP-1, is a neuroendocrine hormone secreted by the small intestine. Hormone receptors are present not only in regulatory areas of the brain but also in the pancreas. Both regulatory areas contribute to blood sugar control. Activating GLP-1 receptors can promote insulin secretion and inhibit glucagon secretion in a glucose concentration-dependent manner, thereby increasing satiety, inhibiting food intake, and delaying gastric emptying, ultimately achieving the goals of regulating blood sugar and weight loss.

[0004] Although semaglutide is favored by patients with type 2 diabetes due to its good therapeutic effects, its dosage is still limited by adverse reactions such as nausea and vomiting, and single-target administration usually cannot achieve complete blood glucose control and weight loss. Gastric inhibitory peptides (GIPs) are peptides secreted by K cells in the intestine that can stimulate insulin secretion in a glucose-dependent manner. Furthermore, GIPs can reduce weight by decreasing food intake and increasing energy expenditure. Single-target therapy with GIPs has shown some hypoglycemic effects in type 2 diabetes, but under pathological conditions, GIPs in the body cannot fully function as they do under healthy physiological conditions. Summary of the Invention

[0005] Purpose of the Invention: To address the problems existing in the prior art, the purpose of this invention is to provide a novel type of GIP-R and GLP-1R dual-agonist polypeptide compound or a pharmaceutically acceptable salt thereof. The polypeptide compound of this invention is a dual incretin peptide compound that activates human glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors. It not only has hypoglycemic and weight-loss effects, but also has a longer half-life and higher bioavailability, and can be used to prepare drugs for the treatment or prevention of diabetes or weight-loss drugs.

[0006] The present invention also provides pharmaceutical compositions of the aforementioned GIP-R and GLP-1R dual agonist polypeptide compounds and their pharmaceutical uses.

[0007] Technical solution: The present invention relates to a class of GIP-R and GLP-1R dual-agonist polypeptide compounds or pharmaceutically acceptable salts thereof, wherein the amino acid sequence of the GIP-R and GLP-1R dual-agonist polypeptide compound is: Y1-X1-EGTFTSDYSI-X2-LDKI-X3-QB n -X4-X5-VQWLI-X6-GGPSSG-X7-PPPS, and the C-terminal amino acid of the GIP-R and GLP-1R dual-activator peptide compound is amidated;

[0008] in,

[0009] Y1 is selected from tyrosine residues or histidine residues;

[0010] X1 is selected from α-aminoisobutyric acid residues;

[0011] X2 is selected from α-aminoisobutyric acid residues, 2-amino-3,3,3-trifluoro-2-methylpropionic acid residues, 1-aminocyclobutanecarboxylic acid residues, or isovaleine residues;

[0012] X3 is selected from α-aminoisobutyric acid residues or alanine residues;

[0013] B nIt is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. n The chemical formula of a -CO2H modified lysine residue, where n is a natural number from 16 to 22, is shown below:

[0014]

[0015] X4 is selected from α-aminoisobutyric acid residues, alanine residues, 2-aminobutyric acid residues, or valine residues;

[0016] X5 is selected from phenylalanine residues, 3-(1-naphthyl)-alanine residues, 3-chlorophenylalanine residues, 3-trifluoromethylphenylalanine residues, 3-fluorophenylalanine residues, 2-fluorophenylalanine residues, 2-chlorophenylalanine residues, 2-trifluoromethylphenylalanine residues, or 3-(2-naphthyl)-alanine residues;

[0017] X6 is selected from α-aminoisobutyric acid residues, serine residues, or alanine residues;

[0018] X7 is selected from α-aminoisobutyric acid residues, 2-aminobutyric acid residues, serine residues, or threonine residues.

[0019] The C-terminal amino acid of the above-mentioned polypeptide compound is amidated to a C-terminal primary amide; specifically, the C-terminus is -CO-NH2.

[0020] Preferably, n = 16, 17, 18, 19, 20, 21, 22. More preferably, n is a natural number from 17 to 22, n is a natural number from 18 to 22, n is a natural number from 19 to 22, n is a natural number from 20 to 22, and n is a natural number from 21 to 22.

[0021] Preferably, the amino acid sequence of the GIP-R and GLP-1R dual-activator polypeptide compound is selected from any of the following:

[0022] (1)SEQ ID NO:1

[0023] YZ-EGTFTSDYSI-Z-LDKI-ZQB 18 -ZF-VQWLI-Z-GGPSSGSPPPS-NH2

[0024] (2)SEQ ID NO:2

[0025] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -ZF-VQWLI-A-GGPSSG-Z-PPPS-NH2

[0026] (3)SEQ ID NO:3

[0027] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AF-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0028] (4)SEQ ID NO:4

[0029] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z4-F-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0030] (5)SEQ ID NO:5

[0031] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z5-F-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0032] (6)SEQ ID NO:6

[0033] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z-Z8-VQWLI-A-GGPSSGAPPPS-NH2

[0034] (7)SEQ ID NO:7

[0035] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z4-Z8-VQWLI-A-GGPSSGAPPPS-NH2

[0036] (8)SEQ ID NO:8

[0037] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0038] (9)SEQ ID NO:9

[0039] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z4-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0040] (10)SEQ ID NO:10

[0041] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0042] (11)SEQ ID NO:11

[0043] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -ZF-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0044] (12)SEQ ID NO:12

[0045] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z4-F-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0046] (13)SEQ ID NO:13

[0047] YZ-EGTFTSDYSI-Z-LDKI-ZQB 18 -AF-VQWLI-Z 14 -GGPPSG-Z-PPPS-NH2

[0048] (14)SEQ ID NO:14

[0049] YZ-EGTFTSDYSI-Z-LDKI-ZQB 18 -AF-VQWLI-A-GGPSSGSPPPS-NH2

[0050] (15)SEQ ID NO:15

[0051] YZ-EGTFTSDYSI-Z-LDKI-ZQB 18 -AF-VQWLI-Z-GGPSSGSPPPS-NH2

[0052] (16)SEQ ID NO:16

[0053] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -Z-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0054] (17)SEQ ID NO:17

[0055] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -Z4-Z8-VQWLI-A-GGPSSG-Z-PPPS-NH2

[0056] (18)SEQ ID NO:18

[0057] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -Z-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0058] (19)SEQ ID NO:19

[0059] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -Z4-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0060] (20)SEQ ID NO:20

[0061] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -A-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0062] (21)SEQ ID NO:21

[0063] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -ZF-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0064] (22)SEQ ID NO:22

[0065] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -Z4-F-VQWLI-Z 14 -GGPSSGAPPPS-NH2

[0066] (23)SEQ ID NO:23

[0067] YZ-EGTFTSDYSI-Z1-LDKI-ZQB 18-AF-VQWLI-A-GGPSSGSPPPS-NH2

[0068] (24)SEQ ID NO:24

[0069] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AF-VQWLI-Z-GGPSSGSPPPS-NH2

[0070] (25)SEQ ID NO:25

[0071] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AF-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0072] (26)SEQ ID NO:26

[0073] YZ-EGTFTSDYSI-Z2-LDKI-AQB 18 -AF-VQWLI-A-GGPPSSG-Z4-PPPS-NH2

[0074] (27)SEQ ID NO:27

[0075] YZ-EGTFTSDYSI-Z2-LDKI-AQB 18 -AF-VQWLI-Z 14 -GGPPSG-Z4-PPPS-NH2

[0076] (28)SEQ ID NO:28

[0077] YZ-EGTFTSDYSI-Z2-LDKI-AQB 18 -A-Z8-VQWLI-A-GGPPSSG-Z4-PPPS-NH2

[0078] (29)SEQ ID NO:29

[0079] YZ-EGTFTSDYSI-Z3-LDKI-AQB 18 -AF-VQWLI-A-GGPSSGSPPPS-NH2

[0080] (30)SEQ ID NO:30

[0081] YZ-EGTFTSDYSI-Z3-LDKI-AQB 18-A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0082] (31)SEQ ID NO:31

[0083] YZ-EGTFTSDYSI-Z3-LDKI-AQB 18 -AF-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0084] (32)SEQ ID NO:32

[0085] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AF-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0086] (33)SEQ ID NO:33

[0087] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -A-Z6-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0088] (34)SEQ ID NO:34

[0089] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -A-Z7-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0090] (35)SEQ ID NO:35

[0091] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0092] (36)SEQ ID NO:36

[0093] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -A-Z9-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0094] (37)SEQ ID NO:37

[0095] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AZ10 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0096] (38)SEQ ID NO:38

[0097] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AZ 11 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0098] (39)SEQ ID NO:39

[0099] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AZ 12 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0100] (40)SEQ ID NO:40

[0101] YZ-EGTFTSDYSI-Z1-LDKI-AQB 18 -AZ 13 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0102] (41)SEQ ID NO:41

[0103] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z6-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0104] (42)SEQ ID NO:42

[0105] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z7-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0106] (43)SEQ ID NO:43

[0107] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0108] (44)SEQ ID NO:44

[0109] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z9-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0110] (45)SEQ ID NO:45

[0111] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 10 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0112] (46)SEQ ID NO:46

[0113] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 11 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0114] (47)SEQ ID NO:47

[0115] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 12 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0116] (48)SEQ ID NO:48

[0117] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 13 -VQWLI-A-GGPSSG-Z-PPPS-NH2

[0118] (49)SEQ ID NO:49

[0119] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AF-VQWLI-Z-GGPSSGSPPPS-NH2

[0120] (50)SEQ ID NO:50

[0121] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z6-VQWLI-Z-GGPSSGSPPPS-NH2

[0122] (51)SEQ ID NO:51

[0123] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z7-VQWLI-Z-GGPSSGSPPPS-NH2

[0124] (52)SEQ ID NO:52

[0125] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z8-VQWLI-Z-GGPSSGSPPPS-NH2

[0126] (53)SEQ ID NO:53

[0127] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z9-VQWLI-Z-GGPSSGSPPPS-NH2

[0128] (54)SEQ ID NO:54

[0129] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 10 -VQWLI-Z-GGPSSG-Z-PPPS-NH2

[0130] (55)SEQ ID NO:55

[0131] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 11 -VQWLI-Z-GGPSSG-Z-PPPS-NH2

[0132] (56)SEQ ID NO:56

[0133] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 12 -VQWLI-Z-GGPSSG-Z-PPPS-NH2

[0134] (57)SEQ ID NO:57

[0135] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 13 -VQWLI-Z-GGPSSG-Z-PPPS-NH2

[0136] (58)SEQ ID NO:58

[0137] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AF-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0138] (59)SEQ ID NO:59

[0139] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z6-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0140] (60)SEQ ID NO:60

[0141] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z7-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0142] (61)SEQ ID NO:61

[0143] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z8-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0144] (62)SEQ ID NO:62

[0145] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z9-VQWLI-Z 14 -GGPSSGSPPPS-NH2

[0146] (63)SEQ ID NO:63

[0147] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 10 -VQWLI-Z 14 -GGPPSG-Z-PPPS-NH2

[0148] (64)SEQ ID NO:64

[0149] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ11 -VQWLI-Z 14 -GGPPSG-Z-PPPS-NH2

[0150] (65)SEQ ID NO:65

[0151] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 12 -VQWLI-Z 14 -GGPPSG-Z-PPPS-NH2

[0152] (66)SEQ ID NO:66

[0153] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AZ 13 -VQWLI-Z 14 -GGPPSG-Z-PPPS-NH2

[0154] (67)SEQ ID NO:67

[0155] HZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0156] (68)SEQ ID NO:68

[0157] YZ-EGTFTSDYSI-Z-LDKI-AQB 16 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0158] (69)SEQ ID NO:69

[0159] YZ-EGTFTSDYSI-Z-LDKI-AQB 17 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0160] (70)SEQ ID NO:70

[0161] YZ-EGTFTSDYSI-Z-LDKI-AQB 19 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0162] (71)SEQ ID NO:71

[0163] YZ-EGTFTSDYSI-Z-LDKI-AQB 20 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0164] (72)SEQ ID NO:72

[0165] YZ-EGTFTSDYSI-Z-LDKI-AQB 21 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0166] (73)SEQ ID NO:73

[0167] YZ-EGTFTSDYSI-Z-LDKI-AQB 22 -A-Z8-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0168] (74)SEQ ID NO:74

[0169] HZ-EGTFTSDYSI-Z-LDKI-AQB 18 -AF-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0170] (75)SEQ ID NO:75

[0171] YZ-EGTFTSDYSI-Z-LDKI-AQB 16 -A-Z7-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0172] (76)SEQ ID NO:76

[0173] YZ-EGTFTSDYSI-Z-LDKI-AQB 17 -A-Z7-VQWLI-A-GGPSSGSPPPS-NH2

[0174] (77)SEQ ID NO:77

[0175] YZ-EGTFTSDYSI-Z-LDKI-AQB 19 -A-Z7-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0176] (78)SEQ ID NO:78

[0177] YZ-EGTFTSDYSI-Z-LDKI-AQB20 -A-Z7-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0178] (79)SEQ ID NO:79

[0179] YZ-EGTFTSDYSI-Z-LDKI-AQB 16 -A-Z9-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0180] (80)SEQ ID NO:80

[0181] YZ-EGTFTSDYSI-Z-LDKI-AQB 17 -A-Z9-VQWLI-A-GGPSSGSPPPS-NH2

[0182] (81)SEQ ID NO:81

[0183] YZ-EGTFTSDYSI-Z-LDKI-AQB 19 -A-Z9-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0184] (82)SEQ ID NO:82

[0185] YZ-EGTFTSDYSI-Z-LDKI-AQB 20 -A-Z9-VQWLI-A-GGPPSSG-Z4-PPPS-NH2

[0186] (83)SEQ ID NO:83

[0187] YZ-EGTFTSDYSI-Z-LDKI-AQB 16 -A-Z6-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0188] (84)SEQ ID NO:84

[0189] YZ-EGTFTSDYSI-Z-LDKI-AQB 17 -A-Z6-VQWLI-A-GGPPSSG-Z-PPPS-NH2

[0190] (85)SEQ ID NO:85

[0191] YZ-EGTFTSDYSI-Z-LDKI-AQB 19-A-Z6-VQWLI-A-GGPSSG-Z-PPPS-NH2

[0192] (86)SEQ ID NO:86

[0193] YZ-EGTFTSDYSI-Z-LDKI-AQB 20 -A-Z6-VQWLI-A-GGPSSG-Z-PPPS-NH2

[0194] (87)SEQ ID NO:87

[0195] YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -A-Z8-VQWLI-A-GGPSSG-Z4-PPPS-NH2

[0196] In the above polypeptide sequence: Z is an α-aminoisobutyric acid residue; Z1 is a 2-amino-3,3,3-trifluoro-2-methylpropionic acid residue; Z2 is a 1-aminocyclobutanecarboxylic acid residue; Z3 is an isovale residue; Z4 is a 2-aminobutyric acid residue; Z5 is a valine residue; Z6 is a 3-(1-naphthyl)-alanine residue; Z7 is a 3-chlorophenylalanine residue; Z8 is a 3-trifluoromethylphenylalanine residue; Z9 is a 3-fluorophenylalanine residue; Z 10 It is a 2-fluorophenylalanine residue; Z 11 It is a 2-chlorophenylalanine residue; Z 12 It is a 2-trifluoromethylphenylalanine residue; Z 13 It is a 3-(2-naphthyl)-alanine residue; Z 14 It is a D-type serine residue; B 16 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 16 -CO2H modified lysine residues; B 17 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 17 -CO2H modified lysine residues; B 18 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 18 -CO2H modified lysine residues; B 19 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 19 -CO2H modified lysine residues; B 20It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 20 -CO2H modified lysine residues; B 21 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 21 -CO2H modified lysine residues; B 22 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 22 -CO2H modified lysine residues.

[0197] Preferably, the salt is a salt formed by the GIP-R and GLP-1R dual-activating polypeptide compound and one of the following compounds: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfonic acid, phosphoric acid, nitric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3- Hydroxy-2-naphtholic acid, nicotinic acid, pyruvic acid, pectinic acid, persulfate, 3-phenylpropionic acid, picric acid, pentanoic acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfate, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, ferric acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucohepanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfonic acid, or thiocyanate.

[0198] On the other hand, the present invention provides a pharmaceutical composition comprising the above-described GIP-R and GLP-1R dual agonist polypeptide compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0199] Preferably, the pharmaceutically acceptable carrier is a pharmaceutically acceptable excipient, diluent, buffer, adjuvant, excipient, or medium.

[0200] Preferably, the formulation is in the form of tablets, capsules, elixirs, syrups, lozenges, inhalers, sprays, injections, films, patches, powders, granules, blocks, emulsions, suppositories, or compound preparations.

[0201] On the other hand, the present invention provides the use of the above-mentioned GIP-R and GLP-1R dual agonist polypeptide compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing diseases or conditions caused by or characterized by excessive weight, or in the preparation of a weight-loss medicament.

[0202] Preferably, the disease or symptom is selected from obesity, diabetes, and hyperglycemia.

[0203] Preferably, the disease or symptom is a human disease or symptom.

[0204] Preferably, the use of the GIP-R and GLP-1R dual-agonist polypeptide compound or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof, in the preparation of a medicament for the treatment and / or prevention of diseases or conditions caused by or characterized by excessive weight, or in the preparation of a weight-loss medicament, by administering simultaneously, separately or sequentially in combination effective amounts of one or more of metformin, thiazolidinediones, sulfonylureas, dipeptidyl peptidase 4 inhibitors or sodium-glucose cotransporters.

[0205] Unless otherwise specified, the terms used in this invention generally have the following meanings:

[0206]

[0207] This invention combines the GLP-1 target with other targets to develop a novel dual-target diabetes drug, aiming to improve the efficacy of diabetes treatment while reducing adverse reactions. Based on the physiological synergistic effect between GIP agonists and GLP-1 agonists, this invention combines them to develop new dual-target GIP / GLP-1 drugs, GIP-R and GLP-1R dual-agonist peptide compounds, which produce stronger effects in controlling blood glucose and reducing weight. This also provides a novel medication option and treatment plan for clinical application in patients with type 2 diabetes.

[0208] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0209] This invention provides a novel GIP-R and GLP-1R dual-agonist polypeptide compound or a pharmaceutically acceptable salt thereof, which can be prepared by artificial synthesis, i.e. polypeptide solid-phase synthesis. It has a simple structure and low cost, making it easy to promote and use and industrial production applications.

[0210] Compared with existing semaglutides, the GIP-R and GLP-1R dual-agonist polypeptide compounds of the present invention have a more significant dual-agonist effect on the receptors of human glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1). They not only have hypoglycemic and weight-loss effects, but also have a longer half-life and higher bioavailability. They can avoid adverse reactions such as nausea and vomiting caused by long-term use of existing drugs, and can be used as effective raw materials for the preparation of drugs for the treatment or prevention of diabetes or weight-loss drugs. Detailed Implementation

[0211] Unless otherwise defined, the technical terms used in the following embodiments have the same meanings as commonly understood by those skilled in the art. Unless otherwise specified, the experimental reagents used in the following embodiments are conventional biochemical reagents; and the experimental methods described are conventional methods.

[0212] The present invention will be further described below with reference to specific embodiments.

[0213] This invention modifies existing semaglutide to obtain a novel GIP-R structure.

[0214] GLP-1R dual-activating polypeptide compounds. The polypeptide compounds SEQ ID NO:1-87 in the embodiments of this invention can all be directly synthesized artificially using a polypeptide solid-phase method. The polypeptides designed in this invention were all directly synthesized by a biotechnology company.

[0215] Among them, B n For known residues, see the literature: Discovery of the Once-Weekly Glucagon-Like Peptide-1 (GLP-1) Analogue Semaglutide, J.Med.Chem.2015,58,7370-7380; CN110684082A; where the size of n can be designed and prepared in different ways according to existing methods.

[0216] Example 1

[0217] 1. Experiments on the agonistic activity of peptide compounds on human GIP-R and GLP-1R receptors

[0218] HEK293 cells were co-transfected with cDNA encoding human GIP-R and GLP-1R receptors, respectively. Stable transfected cell lines were selected and cultured in DMEM medium (containing 10% fetal bovine serum FCS, 100 IU / mL penicillin, and 100 μg / mL streptomycin). Semaglutide was used as a positive control. In the assay for peptide compounds, 3 × 10⁻⁶ cDNAs were used. 4 Cells (200 μL / well) were seeded in 96-well plates. Peptide compounds were diluted to different concentrations using culture medium and added to co-transfected HEK293 cells. After 20 min of cell incubation, fluorescence readings were measured using a CisBio cAMP assay kit and a multi-mode microplate reader. A standard curve was established to convert the fluorescence readings into corresponding cAMP values. The EC50 of the peptides was calculated using a non-linear regression method with Graphpad Prism 7.0 software. 50 Numerical values. Experimental results are shown in Table 1. Agonistaltic activity (EC50) of peptide compounds on human GIP-R and GLP-1R receptors. 50 ,nM).

[0219] As shown in Table 1, compared with the positive control drug Semaglutide, all 87 peptide compounds exhibited more than 2-fold agonistic activity against the human GLP-1R receptor (ECG). 50 Simultaneously, it showed more than 100-fold agonistic activity against GIP receptors (ECG). 50 Both were significantly stronger than the positive control drug Semaglutide.

[0220] Table 1 shows the agonistic activity of all 87 tested peptide compounds on human GIP-R and GLP-1R receptors (EC50). 50 ,nM)

[0221]

[0222]

[0223] 2. Effects of dual peptide agonists on body weight, fat intake, blood glucose and triglyceride levels in obese C57BL / 6J DIO mice.

[0224] C57BL / 6J mice (DIO) are particularly well-suited for research or modeling of type 2 diabetes. The effects of the compounds of this invention on weight loss, body composition, and hepatic steatosis were evaluated in C57BL / 6J DIO mice. In this experiment, diet-induced obese male C57BL / 6J DIO mice (23-24 weeks old) were placed in an SPF-grade environment with a standard diet and free access to water. All experimental procedures were performed in accordance with guidelines for laboratory animal ethics and practices. After two weeks of acclimatization to a standard diet, mice were randomly assigned to treatment groups (n=10 / group) based on body weight, with each group having a similar starting average body weight. Negative control excipients, positive control drugs semaglutide (10 nmol / kg and 30 nmol / kg), and peptide compounds (dose range 10–100 nmol / kg) were dissolved in excipients (20 mM citrate buffer, pH 7.0) and administered subcutaneously to free-feeding DIO mice 30–90 minutes before the start of the dark cycle, every three days for 15 days. Daily changes in body weight were measured throughout the study. Absolute changes in body weight were calculated by subtracting the body weight of the same animal prior to the first compound injection. Total fat mass was measured on days 0 and 14 using nuclear magnetic resonance (NMR) on an EchoMRI-100™ instrument (Echo Medical Systems, Houston, TX). On day 15, blood glucose was measured using an Accu-Chek glucometer from the tail vein of mice, followed by sacrifice of the mice and collection of liver homogenate for measurement of liver triglycerides on a Hitachi Modular P analyzer.

[0225] Table 2 shows that compared with the negative control group mice, the changes in body weight and fat mass in the peptide group mice were significantly different (****, p<0.0001), indicating that the peptide compound (SEQ ID NO:1-87) of the present invention can effectively reduce body weight and fat mass. Furthermore, compared with semaglutide, the peptide group mice were more effective in reducing body weight and fat mass, with an efficacy improvement of more than 2 times. Therefore, the peptide compound described in this invention is suitable for the development of weight-loss indications. Furthermore, from all 87 polypeptide compounds sharing a common motif structure, 16 representative polypeptides (including SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:43, SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO:61, SEQ ID NO:67, SEQ ID NO:74, and SEQ ID NO:86) were selected for animal treatment experiments in high, medium, and low dose groups. As shown in Table 2, these 16 polypeptide compounds reduced body weight and fat mass in a dose-dependent manner. Further, as shown in Table 3, compared with semaglutide, polypeptide compounds (SEQ ID NO:1-87) can effectively and significantly reduce blood glucose and triglycerides, especially showing significantly better effects in reducing triglycerides. In summary, the polypeptide compound described in this invention has a more significant effect on both lowering blood sugar and reducing weight, and can be applied to the development of drugs for treating diabetes or for weight loss.

[0226] Table 2. Effects of drug administration on body weight and fat mass changes in DIO mice in the experiment (n=10)

[0227]

[0228]

[0229]

[0230]

[0231] SPSS software was used for data analysis in this experiment. All experimental data are expressed as mean ± standard deviation (mean ± SEM). A p-value < 0.0001 was considered statistically significant between the experimental and control groups.

[0232] Table 3 shows the effects of all 87 peptide compounds tested on blood glucose and liver triglycerides in DIO mice.

[0233]

[0234]

[0235]

[0236] SPSS software was used for data analysis in this experiment. All experimental data are expressed as mean ± standard deviation (mean ± SEM). When ***p < 0.001 and ****p < 0.0001, the experimental group was statistically significant compared to the control group.

[0237] Example 2

[0238] In vivo pharmacokinetic studies of the polypeptide compounds of this invention.

[0239] The pharmacokinetic properties of the polypeptide compound of the present invention were evaluated in cynomolgus monkeys. The compound was administered intravenously or subcutaneously at a volume of 0.21 ml / kg in 20 mM citrate buffer (pH 7.0). Blood samples were collected from each animal at 2, 4, 8, 12, 24, 48, 72, 96, 120, 144, 168, 204, 240, and 312 hours after administration. Plasma concentrations of the polypeptide compound were determined by LC / MS after solid-phase extraction. The in vivo half-life (T0) was calculated using Phoenix software. 1 / 2 ) and bioavailability F (%).

[0240] As shown in Table 4 above, compared with semaglutide, the dual-agonist polypeptides (SEQ ID NO: 1-87) provided by this invention have a half-life of over 40 hours in animals. Compared with semaglutide, the polypeptide compounds (SEQ ID NO: 1-87) provided by this invention exhibit a longer half-life and higher bioavailability, thus demonstrating superior long-acting therapeutic effects compared to semaglutide. Therefore, the long-acting dual-agonist polypeptides of this invention are suitable for the development of indications for hypoglycemic and weight-loss treatments.

[0241] Table 4 shows the half-life (T) after a single subcutaneous administration of 0.2 mg / kg to male cynomolgus monkeys. 1 / 2 (hr) and bioavailability F (%)

[0242]

[0243]

[0244] As can be seen from the above embodiments, the dual-agonist polypeptide compound described in this invention is a class of highly efficient dual-agonist polypeptide compounds, exhibiting dual agonist activity (ECG) on human GIP-R and GLP-1R receptors. 50 It is significantly stronger than semaglutide, and also significantly better than semaglutide in lowering blood sugar and reducing weight. In addition, it has a longer half-life and higher bioavailability, and can be used as an effective raw material to prepare drugs for the treatment or prevention of diabetes or weight loss drugs.

[0245] Finally, it should be noted that other methods can be used to implement this invention. Accordingly, the embodiments of this invention are described as examples, but are not limited to the content described herein, and may include modifications made within the scope of this invention or equivalent content added to the claims. All publications or patents cited in this invention are to be used as references in this invention.

[0246] The above description is merely a preferred embodiment of the present invention and is illustrative only, not intended to limit the scope of protection of the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any changes, modifications, equivalent substitutions, replacements, combinations, simplifications, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A GIP-R and GLP-1R dual-agonist polypeptide compound or a pharmaceutically acceptable salt thereof, characterized in that, The amino acid sequence of the GIP-R and GLP-1R dual-activator polypeptide compound is selected from any of the following: SEQ ID NO:6 YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z-Z8-VQWLI-A-GGPSSGAPPPS-NH2 SEQ ID NO:8 YZ-EGTFTSDYSI-Z-LDKI-AQB 18 -Z-Z8-VQWLI-Z 14 -GGPSSGAPPPS-NH2; In the above polypeptide sequence: Z is an α-aminoisobutyric acid residue; Z8 is a 3-trifluoromethylphenylalanine residue; Z 14 It is a D-type serine residue; B 18 It is ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γGlu)1-CO-(CH2) on the ε-amino side chain. 18 -CO2H modified lysine residues.

2. The GIP-R and GLP-1R dual-agonist polypeptide compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The salt is a salt formed by the GIP-R and GLP-1R dual-activating polypeptide compound with one of the following compounds: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfonic acid, phosphoric acid, nitric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy -2-Naphthoic acid, nicotinic acid, pyruvic acid, pectinic acid, persulfate, 3-phenylpropionic acid, picric acid, tervaline, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfate, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, ferric acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucohepanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfonic acid, or thiocyanate.

3. Use of the GIP-R and GLP-1R dual agonist polypeptide compound of claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of a weight-loss medicament.

4. A pharmaceutical composition comprising the GIP-R and GLP-1R dual agonist polypeptide compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition according to claim 4, characterized in that, The pharmaceutically acceptable carrier is a pharmaceutically acceptable excipient, diluent, buffer, or adjuvant.

6. The pharmaceutical composition according to claim 4, characterized in that, The formulations are in the form of tablets, capsules, elixirs, syrups, lozenges, inhalers, sprays, injections, films, patches, powders, granules, blocks, emulsions, suppositories, or compound preparations.

7. Use of the pharmaceutical composition of claim 4 in the preparation of a weight-loss drug.

8. Use of the GIP-R and GLP-1R dual agonist polypeptide compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 4, in the preparation of a medicament for treating obesity, diabetes, or hyperglycemia.

9. Use of the GIP-R and GLP-1R dual-agonist polypeptide compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 4, in combination with an effective amount of one or more of metformin, thiazolidinediones, sulfonylureas, dipeptidyl peptidase 4 inhibitors, or sodium-glucose cotransporter-2 inhibitors in the preparation of a weight-loss drug.

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