A novel long-acting polypeptide compound, composition and application thereof

Abstract

The present invention discloses a novel long-acting polypeptide compound, composition and application thereof. The long-acting polypeptide compound of the present invention enhances the stability, activity and hydrolysis resistance of the polypeptide by replacing amino acids at key specific sites. Through a large number of experimental explorations, after the 2nd and / or 13th amino acid sites of the main peptide chain of the polypeptide compound of the present invention are replaced with non-natural amino acids Aib, Iva or Cba, the stability, activity and hydrolysis resistance of the polypeptide molecule are significantly enhanced; the 27th and / or 28th amino acid sites are replaced with L and / or D amino acids, respectively, which will also significantly enhance the activity of the polypeptide molecule. The long-acting polypeptide compound of the present invention has a long half-life and has good pharmacodynamic effects in treating diabetes and reducing body weight.

Description

Technical Field

[0001] The present invention belongs to the field of biochemical technology. Specifically, the present invention relates to a novel long-acting polypeptide compound, composition and application thereof that can be used to treat or prevent diabetes or obesity. Background Art

[0002] Diabetes is a group of clinical syndromes caused by the interaction of genetic and environmental factors, in which the body is in a state of high blood sugar levels for a long time. Diabetes is manifested in absolute or relative insufficiency of insulin secretion and decreased sensitivity of target tissue cells to insulin, accompanied by a series of metabolic disorders such as sugar, protein, fat, water and electrolytes. In 2021, the American Diabetes Association (ADA) divided diabetes into four categories: type I diabetes, type II diabetes, gestational diabetes, and other special types of diabetes. Among them, type I diabetes accounts for 5-10% of all diabetic patients, and type II diabetes accounts for about 90-95% of all diabetic patients. Type I diabetes is caused by autoimmune β-cell destruction, which usually leads to absolute insulin deficiency, including latent autoimmune diabetes in adults (LADA). Patients with type II diabetes gradually lose insulin secretion from β cells on the basis of insulin resistance. This disease was once called "non-insulin-dependent diabetes" or "adult-onset diabetes". Its notable pathophysiological characteristics are a decrease in insulin's ability to regulate glucose metabolism (insulin resistance) accompanied by a decrease in insulin secretion due to defective pancreatic β cell function. This is especially true in the early stages of the disease, when insulin resistance is mainly caused by obesity, dyslipidemia, and an unhealthy lifestyle, resulting in a relative lack of insulin secretion (Report of a WHO Consultation, 1999).

[0003] Diabetes is not easy to cure, and its complications bring great pain to patients, usually requiring lifelong medication to control blood sugar. In the field of diabetes treatment, the development in the past decade has been very rapid. In terms of non-insulin drugs, especially GLP-1 receptor agonists and their analogs have been used in clinical practice, which has greatly changed clinical treatment and treatment outcomes (Frontiers in Endocrinology, 2019, 10: 155). It has been reported that GLP-1 is a pancreatic hormone secreted by intestinal L cells, which has pharmacological effects such as promoting insulin secretion, inhibiting the release of glucagon, stimulating pancreatic B cell regeneration, improving insulin sensitivity and increasing glucose utilization (Modern Medicine and Clinic, 2020.). Clinical studies have shown that patients with type 2 diabetes have impaired "incretin effect", but their effects on promoting insulin secretion and lowering blood sugar are not significantly impaired. Therefore, in the field of diabetes treatment, GLP-1 and its related receptors have been clinically studied as important targets for the treatment of type II diabetes, and have shown strong and broad application prospects in the field of diabetes treatment (Expert Guidance on the Clinical Application of GLP-1 Receptor Agonists. Chinese Journal of Diabetes, 2018, 26(05): 353-361.).

[0004] So far, there are several drugs on the market related to GLP-1 receptor agonists (GLP-1RA) and related multi-target agonists. From the initial twice-daily administration to the recent once-a-week administration, the clinical practice has been over 10 years, and a large amount of clinical evidence has been accumulated for hypoglycemic, cardiovascular benefits and weight loss. With the launch of weekly preparations, it has greatly facilitated patients. At the same time, it has a good effect on the treatment of cardiovascular diseases, which has also led to the recognition of this type of drug in recent years. However, it is worth noting that the currently marketed GLP-1 receptor and related target drugs also show considerable toxic side effects and deficiencies, such as severe gastrointestinal reactions, manifested in nausea and vomiting, and there is still a large room for reduction in the frequency of injection administration, etc. (The Lancet, 2009, 374 (9683): ​​39-47.).

[0005] Ultra-long-acting peptide drug molecule modification technology is the key to research and development in this field, and it is also a bottleneck to be broken through internationally. Therefore, how to further design and improve the hypoglycemic effect of compounds, reduce toxic side effects, and increase the duration of drug effect and / or half-life is still an important technical problem to be solved by technicians in this field, which has great social and clinical needs! Summary of the invention

[0006] In view of the above problems, the object of the present invention is to provide a new type of long-acting polypeptide compound.

[0007] Another object of the present invention is to provide a composition containing the above-mentioned long-acting polypeptide compound.

[0008] Another object of the present invention is to provide the application of the long-acting polypeptide compound.

[0009] According to a specific embodiment of the present invention, the amino acid sequence of the long-acting polypeptide compound is as follows:

[0010] X1-X2-X3-GTFTSDYSI-X13-LDKIAQ-X20-AFVQWL-X27-X28-GGPSSG-X35-PPPS-R 1 ;

[0011] wherein, X3 is selected from E, Q or N; X27 is selected from L or I; X28 is selected from A or D; X35 is selected from A or Aib; X20 is K, K(G x (SG) Z -γGlu-CO(CH 2 ) a CO 2 H)、K((PEG 2 ) b -γGlu-CO(CH 2 ) c CO 2 H) or K((AEEA) d -γGlu-CO(CH 2 ) e CO 2 H), wherein x is an integer of 0-5, z is an integer of 1-5, a is an integer of 12-20, b is an integer of 1-8, c is an integer of 12-20, d is an integer of 1-8, and e is an integer of 12-20; R 1 Selected from OH or NH 2 ;

[0012] When X2 is selected from Aib, Iva or Cba: X1 is selected from H or Y; X13 is selected from Aib, Iva or Cba;

[0013] When X2 is D-Ser: X1 is H; X13 is selected from Aib, Iva or Cba.

[0014] Preferably, when X20 is selected from K(G x (SG) Z -γGlu-CO(CH 2 ) a CO 2 H),

[0015] K((PEG 2 ) b -γGlu-CO(CH 2 ) c CO2 H) or K((AEEA) d -γGlu-CO(CH 2 ) e CO 2 H),

[0016] G x (SG) Z -γGlu-CO(CH 2 ) a CO 2 H, (PEG 2 ) b -γGlu-CO(CH 2 ) c CO 2 H or (AEEA) d -γGlu-CO(CH 2 ) e CO 2 H is the "sidearm" structure of the long-acting polypeptide compound described in the present application, and the "sidearm" structure is connected to the main peptide chain of the long-acting polypeptide compound by forming an amide bond with the side chain amino group of the amino acid K on the main peptide chain of the long-acting polypeptide compound.

[0017] Preferably, the 9th amino acid D and the 16th amino acid K on the long-acting polypeptide compound are connected via an amide bond.

[0018] Preferably, in the long-acting polypeptide compound, X20 is K(G x (SG) Z -γGlu-CO(CH 2 ) a CO 2 H), wherein x is 2, z is 2 or 3, and a is 16 or 18.

[0019] Preferably, in the long-acting polypeptide compound, X20 is K((PEG 2 ) b -γGlu-CO(CH 2 ) c CO 2 H), wherein b is 2 and c is 16 or 18.

[0020] Preferably, in the long-acting polypeptide compound, X20 is K((AEEA) d -γGlu-CO(CH 2 ) e CO 2 H), wherein d is 2 and e is 16 or 18.

[0021] Preferably, in the long-acting polypeptide compound, X35 is A.

[0022] Preferably, in the long-acting polypeptide compound, X1 is Y, X2 is selected from Aib, Iva or Cba, X3 is E, and X13 is selected from Aib, Iva or Cba.

[0023] Preferably, in the long-acting polypeptide compound, X2 is Iva, and X13 is Aib or Iva; or, X2 is Aib, and X13 is Iva.

[0024] Preferably, in the long-acting polypeptide compound, X1 is H, X2 is selected from Aib, Iva or Cba, X3 is selected from E or Q; and X13 is selected from Aib, Iva or Cba.

[0025] Preferably, in the long-acting polypeptide compound, X2 is Aib or Iva, and X13 is Iva.

[0026] Preferably, in the long-acting polypeptide compound, X2 is Aib, and X1 is Y; X3 is E; X13 is selected from Iva or Cba; X27 is selected from L or I; X28 is selected from A or D; and X35 is A.

[0027] Preferably, in the long-acting polypeptide compound, X2 is selected from Iva or Cba, and X1 is Y; X3 is E; X13 is selected from Aib, Iva or Cba; X27 is selected from L or I; X28 is selected from A or D; and X35 is A.

[0028] Preferably, in the long-acting polypeptide compound, X2 is selected from Aib, Iva or Cba, and X1 is H; X3 is selected from E or Q; X13 is selected from Aib, Iva or Cba; X27 is selected from L or I; X28 is selected from A or D; and X35 is A.

[0029] Preferably, in the long-acting polypeptide compound, X2 is Aib, and X1 is Y; X3 is E; X13 is Iva; X20 is K(G x (SG) Z -γGlu-CO(CH 2 ) a CO 2 H), wherein x is 2, z is 2 or 3, a is 16 or 18; X27 is selected from L or I; X28 is selected from A or D; X35 is A.

[0030] Preferably, in the long-acting polypeptide compound, X2 is Iva, and X1 is Y; X3 is E; X13 is selected from Aib or Iva; X20 is K(G x (SG) Z -γGlu-CO(CH 2 ) a CO 2H), wherein x is 2, z is 2 or 3, a is 16 or 18; X27 is selected from L or I; X28 is selected from A or D; X35 is A.

[0031] Preferably, in the long-acting polypeptide compound, X2 is selected from Aib or Iva, and X1 is H; X3 is selected from E or Q; X13 is Iva; X20 is K(G x (SG) Z -γGlu-CO(CH 2 ) a CO 2 H), wherein x is 2, z is 2 or 3, a is 16 or 18; X27 is selected from L or I; X28 is selected from A or D; X35 is A.

[0032] Preferably, the long-acting polypeptide compound is selected from any one of the following compounds:

[0033] Compound 1 (SEQ ID NO.1):

[0034] Y-Aib-QGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGPSSG-Aib-PPPS-OH;

[0035] Compound 2 (SEQ ID NO. 2):

[0036] Y-Aib-QGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGPSSGAPPPS-OH;

[0037] Compound 3 (SEQ ID NO.3):

[0038] Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGPSSGAPPPS-OH;

[0039] Compound 4 (SEQ ID NO.4):

[0040] Y-Iva-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPS-OH;

[0041] Compound 5 (SEQ ID NO.5):

[0042] Y-Aib-EGTFTSDYSI-Iva-LDKIAQKAFVQWLIAGGPSSGAPPPS-OH;

[0043] Compound 6 (SEQ ID NO.6):

[0044] Y-Iva-EGTFTSDYSI-Iva-LDKIAQKAFVQWLIAGGPSSGAPPPS-OH;Compound 7 (SEQ ID NO. 7):

[0045] Y-Iva-EGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH; compound 8(SEQ IDNO.8):

[0046] Y-Aib-EGTFTSDYSI-Iva-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH; compound 9(SEQ IDNO.9):

[0047] Y-Iva-EGTFTSDYSI-Iva-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH;Compound 10 (SEQ ID NO. 10):

[0048] Y-Iva-QGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH; compound 11(SEQ IDNO.11):

[0049] Y-Cba-EGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH; compound 12(SEQ IDNO.12):

[0050] Y-Cba-EGTFTSDYSI-Iva-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH;Compound 13 (SEQ ID NO. 13):

[0051] Y-Aib-EGTFTSDYSI-Cba-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH; compound 14(SEQ ID NO.14):

[0052] Y-Iva-EGTFTSDYSI-Cba-LDKIAQKAFVQWLLDGGPSSGAPPPS-OH;Compound 15 (SEQ ID NO. 15):

[0053] H-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLLAGGPSSGAPPPS-OH; compound 16(SEQ IDNO.16):

[0054] H-Iva-QGTFTSDYSI-Aib-LDKIAQKAFVQWLLAGGPSSGAPPPS-OH; Compound 17 (SEQ ID NO. 17):

[0055] H-Iva-EGTFTSDYSI-Iva-LDKIAQKAFVQWLLAGGGPSSGAPPPS-OH;

[0056] Compound 18 (SEQ ID NO.18):

[0057] H-Iva-QGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGPSSGAPPPS-OH;

[0058] Compound 19 (SEQ ID NO.19):

[0059] H-Iva-EGTFTSDYSI-Iva-LDKIAQKAFVQWLLDGPSSGAPPPS-OH;

[0060] Compound 20 (SEQ ID NO.20):

[0061] H-Cba-QGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPS-OH;

[0062] Compound 21 (SEQ ID NO.21):

[0063] H-Cba-QGTFTSDYSI-Iva-LDKIAQKAFVQWLLDGPSSGAPPPS-OH;

[0064] Compound 22 (SEQ ID NO.22):

[0065] H-(D-Ser)-QGTFTSDYSI-Aib-LDKIAQKAFVQWLLDGPSSGAPPPS-OH;

[0066] Compound 23 (SEQ ID NO.23):

[0067] Y-Iva-EGTFTS (D) YSI-Iva-LD (K) IAQKAFVQWLIAGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K through an amide bond;

[0068] Compound 24 (SEQ ID NO.24):

[0069] Y-Aib-EGTFTS (D) YSI-Iva-LD (K) IAQKAFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K through an amide bond;

[0070] Compound 25 (SEQ ID NO.25):

[0071] Y-Iva-EGTFTS (D) YSI-Iva-LD (K) IAQKAFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K through an amide bond;

[0072] Compound 26 (SEQ ID NO.26):

[0073] H-Aib-QGTFTS (D) YSI-Aib-LD (K) IAQKAFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K through an amide bond;

[0074] Compound 27 (SEQ ID NO.27):

[0075] H-Iva-EGTFTS (D) YSI-Aib-LD (K) IAQKAFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K through an amide bond;

[0076] Compound 28 (SEQ ID NO.28):

[0077] Y-Aib-QGTFTSDYSI-Aib-LDKIAQK(GGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSG-Aib-PPPS-NH 2 ;

[0078] Compound 29 (SEQ ID NO.29):

[0079] Y-Aib-QGTFTSDYSI-Aib-LDKIAQK(GGSGSG-γGlu-CO(CH 2 ) 18 CO 2H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0080] Compound 30 (SEQ ID NO.30):

[0081] Y-Aib-EGTFTSDYSI-Aib-LDKIAQK(GGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0082] Compound 31 (SEQ ID NO.31):

[0083] Y-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0084] Compound 32 (SEQ ID NO.32):

[0085] Y-Iva-EGTFTSDYSI-Aib-LDKIAQK(PEG 2 -PEG 2 -γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0086] Compound 33 (SEQ ID NO.33):

[0087] Y-Aib-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0088] Compound 34 (SEQ ID NO.34):

[0089] Y-Aib-EGTFTSDYSI-Iva-LDKIAQK(PEG 2 -PEG 2-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0090] Compound 35 (SEQ ID NO.35):

[0091] Y-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0092] Compound 36 (SEQ ID NO.36):

[0093] Y-Iva-EGTFTSDYSI-Iva-LDKIAQK(AEEA-AEEA-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0094] Compound 37 (SEQ ID NO.37):

[0095] Y-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0096] Compound 38 (SEQ ID NO.38):

[0097] Y-Aib-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0098] Compound 39 (SEQ ID NO.39):

[0099] Y-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0100] Compound 40 (SEQ ID NO.40):

[0101] Y-Iva-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0102] Compound 41 (SEQ ID NO.41):

[0103] Y-Cba-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0104] Compound 42 (SEQ ID NO.42):

[0105] Y-Cba-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0106] Compound 43 (SEQ ID NO.43):

[0107] Y-Aib-EGTFTSDYSI-Cba-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0108] Compound 44 (SEQ ID NO.44):

[0109] Y-Iva-EGTFTSDYSI-Cba-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 ;

[0110] Compound 45 (SEQ ID NO.45):

[0111] H-Aib-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLAGGPSSGAPPPS-OH;

[0112] Compound 46 (SEQ ID NO.46):

[0113] H-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLAGGPSSGAPPPS-OH;

[0114] Compound 47 (SEQ ID NO.47):

[0115] H-Iva-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLAGGPSSGAPPPS-OH;

[0116] Compound 48 (SEQ ID NO.48):

[0117] H-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLAGGPSSGAPPPS-OH;

[0118] Compound 49 (SEQ ID NO.49):

[0119] H-Iva-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-OH;

[0120] Compound 50 (SEQ ID NO.50):

[0121] H-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-OH;

[0122] Compound 51 (SEQ ID NO.51):

[0123] H-Cba-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-OH;

[0124] Compound 52 (SEQ ID NO.52):

[0125] H-Cba-QGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-OH;

[0126] Compound 53 (SEQ ID NO.53):

[0127] H-Aib-QGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSG-Aib-PPPS-OH;

[0128] Compound 54 (SEQ ID NO.54):

[0129] H-(D-Ser)-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2H)AFVQWLLDGGPSSGAPPPS-OH;

[0130] Compound 55 (SEQ ID NO.55):

[0131] Y-Iva-EGTFTS (D) YSI-Iva-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 , wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond;

[0132] Compound 56 (SEQ ID NO.56):

[0133] Y-Aib-EGTFTS (D) YSI-Iva-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 , wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond;

[0134] Compound 57 (SEQ ID NO.57):

[0135] Y-Iva-EGTFTS (D) YSI-Iva-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLLDGGPSSGAPPPS-NH 2 , wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond;

[0136] Compound 58 (SEQ ID NO.58):

[0137] H-Aib-QGTFTS (D) YSI-Aib-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2H) AFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K via an amide bond;

[0138] Compound 59 (SEQ ID NO.59):

[0139] H-Iva-EGTFTS (D) YSI-Aib-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H) AFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is linked to the 16th amino acid K via an amide bond;

[0140] Compound 60 (SEQ ID NO.60):

[0141] Y-Aib-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 .

[0142] The C-terminal -NH 2 It means that the terminal amino acid is amidated to form a C-terminal primary amide, and the -OH at the C-terminus of the compound is the structure of the terminal amino acid itself.

[0143] Preferably, the long-acting polypeptide compound is a pharmaceutically acceptable salt.

[0144] The preparation method of the above series of long-acting polypeptide compounds comprises the following steps:

[0145] Step 1: According to the Fmoc / t-Bu strategy, the main peptide resin corresponding to the main peptide chain of the polypeptide analog is synthesized.

[0146] Step 2: Based on the main peptide resin, according to the Fmoc / t-Bu strategy, the corresponding "side arm" structure is coupled to obtain the corresponding polypeptide resin; wherein the "side arm" structure is PEG 2 -PEG 2 -γGlu-CO(CH 2 ) 18 CO 2 H、AEEA-AEEA-γGlu-CO(CH 2 ) 18 CO 2H、GGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H or GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H.

[0147] Step 3: adding a cleavage solution to the polypeptide resin to perform a cleavage reaction, removing the polypeptide from full protection, extracting a crude compound, and purifying the crude compound.

[0148] According to the preparation method of compounds 31, 35 and 38 of the specific embodiment of the present invention, in step 2, the coupling agents used are 1-hydroxybenzotriazole (HOBt) and N,N-diisopropylcarbodiimide (DIC), the solvent is N,N-dimethylformamide (DMF), and the Fmoc group is removed with 20% piperidine (Piperidine) / N,N-dimethylformamide solution; in step 3, the cleavage solution is trifluoroacetic acid (TFA), 2,2'-(1,2-ethylenedioxy)bis(ethanediol) (DODT), m-cresol, H 2 O is composed in a volume ratio of 92.5:2.5:2.5:2.5; the crude compound extraction method includes filtration, precipitation and / or methyl tert-butyl ether extraction. The purity of the obtained compounds 31, 35, and 38 is greater than 96%.

[0149] Another object of the present invention is to provide a composition, which contains a long-acting polypeptide compound and a pharmaceutically acceptable carrier or excipient. For example, a carrier that can reduce drug degradation and loss and reduce side effects, such as micelles, microemulsions, gels and other carriers; excipients refer to materials added to make the drug into a suitable dosage form, such as buffers, lyophilization excipients, etc., which can make the pharmaceutical composition containing the compound of the present invention into a solution or lyophilized powder for parenteral administration. Before use, the lyophilized powder can be added with an appropriate solvent or other pharmaceutically acceptable carrier to reconfigure the powder. The liquid formula is generally a buffer solution, an isotonic solution and an aqueous solution. The buffer solution can be a phosphate buffer solution, the isotonic solution can be a 0.9% sodium chloride solution, and the aqueous solution is a solution obtained by directly dissolving with purified water.

[0150] Those skilled in the art will appreciate that the pharmaceutical composition prepared by adding the long-acting polypeptide compound as an active ingredient and a pharmaceutically acceptable carrier and / or excipient is suitable for various modes of administration, such as oral administration, transdermal administration, intravenous administration, intramuscular administration, topical administration, nasal administration, etc. According to the mode of administration adopted, the pharmaceutical composition of the present invention can be prepared into various suitable dosage forms, which contain at least one effective dose of the compound of the present invention and at least one pharmaceutically acceptable pharmaceutical carrier. Examples of suitable dosage forms are tablets, capsules, sugar-coated tablets, granules, oral solutions and syrups, ointments and patches for skin surfaces, aerosols, nasal sprays, and sterile solutions that can be used for injection.

[0151] The dosage of the pharmaceutical composition of the present invention can vary within a wide range, and those skilled in the art can determine it based on objective factors, such as the type of disease, severity of the disease, patient weight, dosage form, route of administration, and the like.

[0152] Another object of the present invention is to provide applications of long-acting polypeptide compounds and compositions.

[0153] The present invention obtains a series of long-acting polypeptide compounds, and studies the pharmacodynamic effects of this series of drugs. Studies have shown that the long-acting polypeptide compounds of the present invention have a longer half-life, have insulinotropic activity, have no adverse reactions, can be used to treat diabetes and obesity, and can potentially be used as a new generation of drugs for treating diabetes and obesity.

[0154] The applications of the long-acting polypeptide compound and composition of the present invention specifically include:

[0155] In the preparation of drugs for preventing or treating diabetes, in the preparation of drugs for preventing or treating obesity.

[0156] The innovative results and beneficial effects of the present invention are:

[0157] The solution to the half-life and stability of polypeptides is the key to whether the design of polypeptide drugs can be made into drugs, and it is also a major scientific and core issue studied in this field. Among them, the replacement of amino acids at key specific sites enhances the stability, activity and hydrolysis resistance of the polypeptide. Through a large number of experimental explorations, after the 2nd and / or 13th amino acid sites of the main peptide chain of the polypeptide compound of the present invention are replaced with non-natural amino acids Aib, Iva or Cba, the stability, activity and hydrolysis resistance of the polypeptide molecule are significantly enhanced; the 27th and / or 28th amino acid sites are replaced with L and / or D amino acids, respectively, which will also significantly enhance the activity of the polypeptide molecule. These key discoveries in the development of innovative polypeptide drugs are all based on a large number of experimental explorations (see specific examples). In addition to the innovation of the main peptide chain, the development of ultra-long-acting polypeptide drug molecule modification technology is also key and is a bottleneck to be broken through internationally in this field. The present invention has developed site-specific side chain modification technology through bioinformatics, structural biology, computer-aided design, structure-activity relationship research, etc., breaking through the ultra-long-acting polypeptide and protein drug molecular modification technology, greatly extending the half-life of the synthesized compound, and realizing the ultra-long-acting polypeptide drug. The polypeptide compound involved in the present invention not only has a primary structure, but also has a secondary / tertiary structure that is crucial to the activity of the polypeptide compound. Various proteins / polypeptides have specific secondary / tertiary spatial conformations, and these specific spatial conformations are related to their specific biological functions, and the structure and function are highly unified; the secondary and tertiary spatial structures have a significant impact on whether the compound can bind to the target, how to bind to the target, and the strength of the binding, that is, the secondary / tertiary structure is crucial for the polypeptide compound to exert its biological function. The addition of side chains in the present invention greatly affects the secondary and tertiary spatial structures of the polypeptide compounds. Therefore, in the present invention, the addition of different side chains causes a great change in the spatial conformation of the main peptide chain of the polypeptide compound involved in the present application, and the change in the polypeptide conformation will affect the performance of its biological function. The biological properties of the polypeptide are unpredictable. Whether it is effective for the long-term effect of a specific polypeptide or protein requires a large number of biopharmaceutical tests and experiments to know.

[0158] The design and synthesis of the novel long-acting polypeptide compound described in the present invention and the long-acting modification technology used are only effective for the polypeptide compound described in the present invention or an uncertain polypeptide compound, and are unpredictable. The results of the test (Example 6) to further verify the creativity and novelty of the invention show that the long-acting polypeptide compound of the present invention greatly prolongs the half-life, and the drug half-life in rats can reach more than 22 hours, achieving the ultra-long-acting polypeptide drug, and achieving a frequency of once every 2 weeks or more for human use. The drug half-life of the polypeptide drug in rats currently reported in this field is basically less than 10 hours, and the frequency of drug administration can only be achieved once a week.

[0159] The side chain modification technology of this application is not universally applicable. Whether it is effective for long-term effects on specific peptides or proteins while maintaining activity requires a large number of innovative biopharmaceutical tests and experiments to determine. To illustrate this point, we also synthesized the following control compounds, compound 61 is a similar compound, compound 62 is the marketed peptide GLP-1 drug lixisenatide, and compounds 63 and 64 are compounds obtained by side chain modification of different positions of lixisenatide:

[0160] Compound 61 (SEQ ID NO.61):

[0161] Y-Aib-EGTFTSDYSI-Aib-LDKIAQK(AEEA-AEEA-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2 ;

[0162] Compound 62 (SEQ ID NO.62):

[0163] HGEGFTTSDLSKQMEEEVRLFIEWLKNGGPSSGAPPSKKKKKK-OH;

[0164] Compound 63 (SEQ ID NO.63):

[0165] HGEGFTSDLSK(GGSGSGSG-γGlu-CO(CH 2 )1 8 CO 2 H)QMEEEVRLFIEWLKNGGPSSGAPPSKKKKKK-OH;

[0166] Compound 64 (SEQ ID NO.64):

[0167] HGEGFTTSDLSKQMEEEVRLFIEWLK(GGSGSGSG-γ-Glu-CO(CH 2 ) 18 CO 2 H)NGGPSSGAPPSKKKKKK-OH;

[0168] The C-terminal -NH 2 It means that the terminal amino acid is amidated, and the -OH at the C-terminus of compounds 62-64 is the structure of the terminal amino acid itself.

[0169] Our research found that lixisenatide can bind to the extracellular domain and transmembrane pocket of GLP-1R, and form a suitable steric hindrance with the GLP-1R receptor. However, the two modified peptides (compounds 63 and 64) have a change in conformation due to the entanglement of the long-acting side chain, thereby forming a large steric hindrance with GLP-1R, hindering ligand-receptor binding, and only showing entanglement with the extracellular domain of GLP-1R, but there is no binding site in the transmembrane pocket, which not only fails to prolong its hypoglycemic effect, but also causes the original parent peptide to lose its hypoglycemic effect. The results of our activity test (Example 5) show that lixisenatide cannot improve glucose tolerance by relying solely on side chain modification, and the hypoglycemic effect is greatly reduced or even inactivated. Therefore, the compounds with new activity obtained by the present invention through independent design, modification, synthesis, and screening through a series of activity verification tests in cell models and animal models are original.

[0170] The novel long-acting polypeptide compound of the present invention utilizes lipophilic substituents to bind to albumin in the blood, protecting it from enzymatic degradation, thereby increasing the half-life. The helical structure of the molecule is stabilized by an intramolecular bridge, thereby increasing the efficacy and / or selectivity against the target.

[0171] The novel long-acting polypeptide compound of the present invention has high synthesis yield, good stability, is easy to scale up production and has low cost.

[0172] At the same time, the novel long-acting polypeptide compound of the present invention has a better pharmacological effect of reducing body weight. The long-acting polypeptide compound can be used to prevent weight gain or promote weight loss by causing a decrease in food intake and / or an increase in energy consumption. Therefore, the novel long-acting polypeptide compound of the present invention can also be used to directly or indirectly treat other diseases caused by or characterized by overweight, such as treating and / or preventing obesity, morbid obesity, obesity-related inflammation, obesity-related gallbladder disease, and sleep apnea caused by obesity. The effect of the present invention in these diseases may be due to the direct or indirect effect of the novel long-acting polypeptide compound on body weight, or the effect on other aspects of the body other than body weight.

[0173] The specific meanings of the abbreviations used in the present invention are as follows:

[0174] DCM is dichloromethane; DMF is N, N-dimethylformamide; MeOH is methanol; Piperidine is piperidine; HOBt is 1-hydroxybenzotriazole; DIC is N, N'-diisopropylcarbodiimide; Fmoc is fluorenylmethoxycarbonyl; resin is resin; FBS is fetal bovine serum; H and His are histidine; Y and Tyr are tyrosine; E and Glu are glutamic acid; Q and Gln are glutamine; N and Asn are asparagine; G and Gly are glycine; T and Th are r is threonine; F and Phe are phenylalanine; S and Ser are serine; D and Asp are aspartic acid; I and IIe are isoleucine; L and Leu are leucine; K and Lys are lysine; A and Ala are alanine; V and Val are valine; W and Trp are tryptophan; P and Pro are proline; Aib is 2-aminoisobutyric acid; Iva (Isovaline) is isovaline; Cba (1-Aminocyclobutanecarboxylic acid) is α-aminocyclobutanecarboxylic acid; Alloc is allyloxycarbonyl; PEG 2 is 3-oxo-2,7,10-trioxa-4-azatridecan-13-oic acid, AEEA is 8-amino-3,6-dioxaoctanoic acid; TFA is trifluoroacetic acid; DODT is 2,2'-(1,2-ethylenedioxy)bis(ethanediol); and ACN is acetonitrile. BRIEF DESCRIPTION OF THE DRAWINGS

[0175] Figure 1 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 2, 0.5 h after administration to mice;

[0176] Figure 2 for Figure 1 Area under the blood glucose curve (AUC);

[0177] Figure 3 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 2, 1 hour after administration to mice;

[0178] Figure 4 for Figure 3 Area under the blood glucose curve (AUC);

[0179] Figure 5 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 24 hours after administration to mice;

[0180] Figure 6 for Figure 5 Area under the blood glucose curve (AUC);

[0181] Figure 7 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 for mice 48 hours after administration.

[0182] Figure 8 for Figure 7 Area under the blood glucose curve (AUC);

[0183] Fig. 9 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 72 hours after administration to mice.

[0184] Fig.10 for Fig. 9 Area under the blood glucose curve (AUC);

[0185] Fig.11 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 in mice 96 hours after administration.

[0186] Fig.12 for Fig.11 Area under the blood glucose curve (AUC);

[0187] Fig.13 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 120 hours after administration to mice.

[0188] Fig.14 for Fig.13 Area under the blood glucose curve (AUC);

[0189] Fig.15 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 144 hours after administration to mice.

[0190] Fig.16 for Fig.15 Area under the blood glucose curve (AUC);

[0191] Fig.17 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 3 168 hours after administration to mice.

[0192] Fig.18 for Fig.17 Area under the blood glucose curve (AUC);

[0193] Fig.19 This is a statistical graph of the weight monitoring data of mice in Example 4;

[0194] Fig. 20 The fasting blood glucose monitoring data of the mice in Example 4 are statistically analyzed;

[0195] Fig.21 This is a graph showing the time-blood glucose results of the OGTT experiment in Example 5 24 hours after administration to mice;

[0196] Fig. 22 for Fig.21 Area under the blood glucose curve (AUC). DETAILED DESCRIPTION

[0197] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0198] Materials and Methods

[0199] Boc-His(Trt)-OH and Fmoc-Aib-OH were purchased from Shanghai Jier, and mono-tert-butyl eicosanedioate was self-made. The remaining amino acids were purchased from Chengdu Zhengyuan Company, and the condensation agent was purchased from Suzhou Haofan Company. Unless otherwise specified, all other reagents were analytically pure, and the solvents were purchased from Shanghai Titan Company. The centrifuge was purchased from Lu Xiangyi. 5.0 cm reverse phase C 18 A preparative column (46 mm x 250 mm) was used to purify the peptide. The high performance liquid chromatograph was a product of Thermo Fisher Scientific. Mass spectrometry was performed using a Waters mass spectrometer.

[0200] Example 1 Synthesis of polypeptide compounds

[0201] 1. Synthesis of Compound 31

[0202] Amino acid sequence of compound 31:

[0203] Tyr-Iva-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Aib-Leu-Asp-Lys-Ile-Ala-Gln-Lys(Gl y-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γGlu-CO(CH 2 ) 18 CO 2 H)-Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gl y-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH 2

[0204] Abbreviation: Y-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH 2 ) 18 CO 2 H)AFVQWLIAGGPSSGAPPPS-NH 2

[0205] method:

[0206] Step 1. Synthesize the main peptide resin corresponding to the main peptide chain

[0207] According to the Fmoc / t-Bu strategy, the synthesis scale was 0.5 mmol, and the following main peptide resin was synthesized:

[0208] Boc-Tyr(tBu)-Iva-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(t Bu)-Ile-Aib-Leu-Asp(OtBu)-Lys-Ile-Ala-Gln(Trt)-Lys(Alloc)-Ala-Phe-Val-Gln(Trt)-Trp(Bo c)-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Resin.

[0209] (1): Weigh 0.89 g of Rink Amide AM Resin (loading 0.56 mmol / g, Xi'an Lanxiao), add it to the reaction column, add 15 ml of DMF to swell for 30 min, weigh Fmoc-Ser(tBu)-OH: 1.587 g (6 eq), HOBt: 0.672 g (7.2 eq), DMAP: 0.06 g (0.72 eq), and set aside. Draw out DMF, wash the resin twice with DMF, and add the above weighed materials to the reaction column. Add an appropriate amount of DMF, stir evenly with nitrogen, and add DIC 0.83 mL (7.8 eq). React for 2 h, and the reaction is complete. Draw out the reaction solution, wash it three times with DMF, and add acetic anhydride / pyridine (7:6, v / v) to seal for 4 h. The blocking solution was removed and the plate was washed 6 times with DMF to obtain Fmoc-Ser(tBu)-Rink Amide AM Resin.

[0210] (2): Using Fmoc-Ser(tBu)-Rink Amide AM Resin as carrier, HOBt and DIC as coupling agents, and DMF as solvent, the Fmoc group was removed with 20% Piperidine / DMF solution (twice for 5 min + 7 min), and the coupling effect was monitored with ninhydrin during the coupling process. Manual feeding was performed, and condensation reactions were performed from the C-terminus to the N-terminus to connect Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmo c-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmo c-Ile-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmo c-Ala-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH, Fmoc-Ile-OH, Fmoc-Lys(Boc) -OH, Fmoc-Asp(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Aib-OH, Fmoc-Ile-OH, Fmoc-Ser(tBu)-OH, Fmoc-Se r(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmo c-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Gly-OH, Fmoc-Glu(tBu)-OH, Fmoc-Iva-OH, Boc-Tyr(tBu)-OH. The above amino acid feed is equivalent to a synthesis scale of 5 eq, and Boc-Tyr(tBu)-Iva-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys-Ile-Ala-Gln(Trt)-Lys(Alloc)-Ala-Phe-Val-Gln(Trt)-Trp(Boc)-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Resin is obtained.

[0211] There are a few points to note:

[0212] 1) Synthesis of Fmoc-Ser(tBu)-Rink Amide AM Resin: Since the substitution degree of Rink Amide AM Resin itself is low, the amount of Fmoc-Ser(tBu)-OH should be large, otherwise the substitution degree will be low and the material will be wasted. Use acetic anhydride / pyridine to block to prevent the appearance of defective peptides.

[0213] 2) In each subsequent condensation reaction, the amount of Fmoc-protected amino acid, HOBt, and DIC added was 5 times, and the reaction time was 2 hours.

[0214] 3) During the coupling process, the coupling effect was monitored with ninhydrin. If the test result was negative, the reaction was complete. If it was positive, it needed to be repeated. The amount of Fmoc-protected amino acid, HOBt, and DIC was 2 times, and the reaction time was 1 hour. If it was still positive, acetic anhydride / pyridine (7:6, v / v) was added to block for 2 hours.

[0215] (3): Removal of allyloxycarbonyl (Alloc)

[0216] DCM was added to the resin, 0.5 mL (12 eq) of morpholine was added, and 0.173 g of Pd(PPh 3 ) 4 (0.3eq) was added to the reaction column and reacted for 1 hour. After the reaction was completed, the reaction solution was removed and washed with DMF 3 times and DCM 6 times. The main peptide chain peptide resin was obtained: Boc-Tyr(tBu)-Iva-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Ty r(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys-Ile-Ala-Gln(Trt)-Lys-Ala-Phe-Val-Gln(Trt)-Trp(Boc)-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Resin.

[0217] Step 2. Coupling of the "sidearm" structure:

[0218] Coupling Fmoc-Gly-OH: Add Fmoc-Gly-OH, HOBt, and an appropriate amount of DMF to the main peptide resin product, stir evenly with nitrogen, add DIC, stir with nitrogen for 2 hours, and use hydrated ninhydrin to detect the coupling effect. If it is colorless and transparent, the reaction is over. Draw off the reaction solution, wash it with N, N-dimethylformamide (DMF) 3 times, remove the Fmoc group with 20% Piperidine / DMF solution (twice 5min + 7min), wash it with DMF 6 times after removing Fmoc, take a sample for ninhydrin detection, and if it is positive, proceed to the subsequent coupling step.

[0219] Repeat the above operation to sequentially couple Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Gly-OH, Fmoc-Glu-OtBu, and mono-tert-butyl eicosandioate. Get Boc-Tyr(tBu)-Iva-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-As p(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys-Ile-Ala-Gln(Trt)-Lys(Gly-Gly-Ser(tBu)-Gly-Ser(tBu)-Gly-Ser(tBu)-Gly-Glu-OtBu-CO(CH 2 ) 18 CO 2 -tBu)-Ala-Phe-Val-Gln(Trt)-Trp(Boc)-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Resin. Wash with DMF 3 times, DCM 3 times, MeOH 2 times, vacuum dry, and obtain 3.2g of dry polypeptide resin.

[0220] Step 3. Deprotection of peptides

[0221] Lysis buffer: TFA, DODT, m-cresol, H 2 O was prepared in advance in a volume ratio of 92.5:2.5:2.5:2.5 and frozen in the refrigerator for 2 hours.

[0222] The dried peptide resin Boc-Tyr(tBu)-Iva-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-Asp(OtBu)-Lys-Ile-Ala-Gln(Trt)-Lys(Gly-Gly-Ser(tBu)-Gly-Ser(tBu)-Gly-Ser(tBu)-Gly-Glu-OtBu-CO(CH 2 ) 18 CO 2 -tBu)-Ala-Phe-Val-Gln(Trt)-Trp(Boc)-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Resin was added with lysis solution, and the temperature was raised to room temperature, and the lysis reaction was carried out for 3 hours. The filtrate was collected by filtration, and the resin was washed 3 times with a small amount of lysis solution. After the filtrate was combined, it was concentrated under reduced pressure to about 1 / 4 of the original volume, and slowly poured into ice methyl tert-butyl ether under stirring, and the residue in the bottle was washed into methyl tert-butyl ether with a small amount of lysis solution. Let it stand for more than 2 hours until the precipitation is complete. The supernatant was removed, the precipitate was centrifuged, washed 3 times with methyl tert-butyl ether, centrifuged, and the solid was blown dry with nitrogen. The crude compound Tyr-Iva-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Aib-Leu-Asp-Lys-Ile-Ala-Gln-Lys(Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γGlu-CO(CH 2 ) 18 CO 2 H)-Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH 2 The crude product weighed 1.48 g.

[0223] Step 4. Purification of crude compound

[0224] The crude compound obtained in step 3 was dissolved in ACN:H 2 O=1:3(v / v) solution, and passed through 5.0cm reverse phase C 18 Preparative HPLC purification was performed on a 46 mm x 250 mm column packed with 39% ACN / H2 Starting with 29% ACN / H2O (containing 0.1% trifluoroacetic acid), the column was eluted with a gradient (increasing the proportion of ACN at a rate of 0.33% / min) at a flow rate of 80 mL / min for 60 minutes, and the fractions containing the peptide were collected to obtain a sample with an HPLC purity greater than 90%. The HPLC purification was repeated once with 29% ACN / H2O. 2 Starting with 5% ACN (containing 0.1% acetic acid), the column was eluted for 60 minutes with a gradient (increasing ACN ratio of 0.33% / min) and a flow rate of 80 mL / min. The fractions containing the polypeptide were collected and freeze-dried to obtain 440 mg of refined peptide with a purity greater than 98.96% and a total yield of 17%.

[0225] Step 5. Product confirmation

[0226] The separated product peptides were identified by LC-MS and 5% ACN / H 2 O (containing 0.1% formic acid) as the starting point, with a gradient (increasing the proportion of ACN at a rate of 6% / min), a flow rate of 0.4 mL / min, and elution analysis for 15 minutes to determine the target compound 31, [M+H] + Calculated value: 5083.73, [M+3H] 3+ The actual measured value is 1695.20.

[0227] 2. Synthesis of Compound 35

[0228] Since the difference between compound 35 and compound 31 is only that X13 in the sequence of the main peptide chain is different, and X13 in 35 is Iva, the difference between the synthesis steps of the two is that step 1 synthesizes the following main peptide resin different from compound 31:

[0229] Boc-Tyr(tBu)-Iva-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Iva-Leu-Asp(OtBu)-Lys-Ile-Ala-G ln(Trt)-Lys(Alloc)-Ala-Phe-Val-Gln(Trt)-Trp(Boc)-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Resin, the synthesis scale is: 0.5mmol.

[0230] The subsequent side chain coupling and cleavage process of compound 35 was the same as that of compound 31.

[0231] The purification and product confirmation methods of compound 35 were the same as those of compound 31, and 332 mg of refined peptide was obtained with a purity greater than 98.46% and a total yield of 13%. The isolated product was identified by LC-MS and confirmed to be the target compound 35, [M+H] + Calculated value: 5097.76, [M+3H] 3+ The actual measured value is 1699.90.

[0232] 3. Synthesis of Compound 38

[0233] Since the difference between compound 38 and compound 31 lies in the difference in X2, X13, X27, and X28 in the sequence of the main peptide chain, X2 in compound 38 is Aib, X13 is Iva, X27 is Leu, and X28 is Asp, the difference in the synthesis steps of the two is that the resins for synthesizing the main peptide in step 1 are different:

[0234] According to the Fmoc / t-Bu strategy, the following main peptide resin was synthesized manually with a synthesis scale of 0.5 mmol: Boc-Tyr(tBu)-Aib-Glu(tBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Iva-Leu-Asp(OtBu)-Lys-Ile-Ala-Gln(Trt)-Lys(Alloc)-Ala-Phe-Val-Gln(Trt)-Trp(Boc)-Leu-Leu-Asp(OtBu)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink Amide AM Re sin.

[0235] The subsequent side chain coupling and cleavage process of compound 38 was the same as that of compound 31.

[0236] The purification and product confirmation methods of compound 38 were the same as those of compound 31, and 385 mg of refined peptide was obtained with a purity greater than 98.35% and a total yield of 15%. The isolated product was identified by LC-MS and confirmed to be the target compound 38, [M+H] + Calculated value: 5128.69, [M+3H] 3+ The actual measured value is 1709.50.

[0237] Based on the above synthesis steps, purification and product confirmation methods, according to the distinguishing sites in the peptide chain of compound 1-60, the coupling order of the resin in step 1 or step 2 of the compound synthesis is adjusted, and the corresponding target product is finally synthesized, and then the separated product is identified by liquid chromatography-mass spectrometry, and the m / z value of the protonated molecular ion peak (measured value in Table 1) is confirmed, and the measured value is compared with the theoretical molecular weight value to confirm that the synthesized and purified product is the target product. Table 1 shows the theoretical molecular weight value, measured value of liquid chromatography-mass spectrometry, sequence and molecular formula of compound 1-60, respectively.

[0238] Table 1 Amino acid sequence of long-acting peptide compounds and LC-MS identification results

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246] Example 2 Effects of Compound 1-27 and Semaglutide on Glucose Tolerance in C57BL / 6J Mice

[0247] We investigated the in vivo efficacy of compound 1-27 and studied the effects and maintenance time of compound 1-27 and semaglutide at the same dose on glucose tolerance in normal mice through an oral glucose tolerance test (OGTT).

[0248] Experimental methods: 8-week-old C57BL / 6J male mice (purchased from Guangdong Sijia Jingda Biotechnology Co., Ltd.) were used in this experiment, 7 mice / group, and the dosage of all compounds and Semaglutide (injection, purchased from Guangzhou Tonghui Pharmaceutical Co., Ltd.) was 50μg / kg. First, the weight and random blood glucose of mice were measured before the experiment, and the mice were regrouped according to the weight and random blood glucose to ensure that the average weight and random blood glucose of each group were similar. The drugs were prepared on the day of administration, and the corresponding drugs were subcutaneously injected according to the group. OGTT tests were performed 0.5h and 1h after administration (parallel experiments, one group of mice underwent 0.5h OGTT test, and another group of mice underwent 1h OGTT test). Glucose was administered by gavage at a dose of 2g / kg, and blood was collected from the tail vein to detect blood glucose values ​​at 0, 15, 30, 60, 90 and 120min after gavage. The data were processed using the software GraphPadPrism, and the time-blood glucose curve was plotted to calculate the area under the blood glucose curve AUC. One-way ANOVA analysis was performed with the PBS control group without medication to calculate the significant difference. Figure 1-4 As shown, * indicates p < 0.05; ** indicates p < 0.01; *** indicates p < 0.001; **** indicates p < 0.0001.

[0249] Result analysis: Figure 1-2 It can be seen that at 0.5h after administration, compared with PBS at each blood sampling time point, all drug-administered groups significantly improved glucose tolerance and had a significant hypoglycemic effect (****, i.e., P≤0.0001), and there was no significant difference between the groups.

[0250] Depend on Figure 3-4 It can be seen that, 1 hour after administration, compared with PBS, compounds 4-14, compounds 16-21 and compounds 23-27 can still improve glucose tolerance and have a significant hypoglycemic effect (****, i.e., P≤0.0001), but the AUC value increased and the hypoglycemic effect was weaker than 0.5 hours after administration. 1 hour after administration, compared with PBS, compounds 1-3, compounds 15 and compounds 22 had no statistically significant difference and lost their hypoglycemic effect.

[0251] Example 3: Effects of Compounds 28-61 and Semaglutide on Glucose Tolerance in C57BL / 6J Mice

[0252] We investigated the in vivo efficacy of compounds 28-60 and used compound 61 as the control group. We conducted an oral glucose tolerance test (OGTT) to study the effects and maintenance time of compounds 28-61 and Semaglutide at the same dose on glucose tolerance in normal mice.

[0253] Experimental methods: This experiment used 8-week-old C57BL / 6J male mice (purchased from Guangdong Sijia Jingda Biotechnology Co., Ltd.), 7 mice / group, and the dosage of all compounds and Semaglutide (injection, purchased from Guangzhou Tonghui Pharmaceutical Co., Ltd.) was 80μg / kg. First, the weight and random blood glucose of mice were measured before the experiment, and the mice were regrouped according to the weight and random blood glucose to ensure that the average weight and random blood glucose of each group were similar. The drugs were prepared on the day of administration and the corresponding drugs were subcutaneously injected according to the group. Glucose OGTT test was performed 24h, 48h, 72h, 96h, 120h, 144h, and 168h after administration. Glucose was gavaged at a dose of 2g / kg, and blood was collected from the tail vein to detect the blood glucose values ​​at 0, 15, 30, 60, 90 and 120min after gavage. The data were processed using the software GraphPadPrism, the time-blood glucose curve was drawn, and the area under the blood glucose curve AUC was calculated. One-way ANOVA analysis was performed with the PBS control group without medication to calculate the significant difference. Figure 5-18 As shown, * indicates p < 0.05; ** indicates p < 0.01; *** indicates p < 0.001; **** indicates p < 0.0001.

[0254] Result analysis:

[0255] (1) 24h results. Figure 5-6 It can be seen that all compounds significantly improved glucose tolerance and lowered blood sugar compared to the PBS control group. Among them, compounds 31-36, compounds 38-39, 41-52, compounds 55-60 and Semaglutide had the most significant blood sugar lowering effects.

[0256] (2) 48h results. Figure 7-8 It can be seen that all compounds have significantly improved glucose tolerance and reduced blood sugar compared with the PBS control group. Among them, compounds 31, 33, 35, compounds 38-39, compounds 41-52 and compounds 55-60 have the best effects, close to 24h; compounds 53 and 54 are second, followed by compounds 37, 40, and 61; and finally compounds 29 and 30; compounds 28, 32, 34, 36 and Semaglutide have similar effects, and their efficacy is lower than 24h.

[0257] (3) 72h results. Figure 9-10 It can be seen that compounds 31, 33, 35, 38, and compounds 39-60 still have good efficacy, slightly weaker than the 48h efficacy; compounds 29, 30, and 61 have the second best efficacy, and compared with the PBS control group, compounds 28, 32, 34, 36 and Semaglutide have no significant difference and have lost their efficacy.

[0258] (4) 96h results. Figure 11-12 It can be seen that compounds 38, 39, 43, 44, 55, 58, 59, and 60 still have efficacy, but are weaker than the 72h efficacy; compounds 31, 33, 35, 37, 40, 49, and 50 have the second best efficacy; followed by compounds 51, 52, 53, and 54; and finally compounds 29 and 30; compared with the PBS control group, compounds 28, 32, 34, 36, 41, 42, 45-48, 53, 56-57, 61 and Semaglutide have no significant differences and have lost their efficacy.

[0259] (5) 120h results. Figure 13-14 It can be seen that compounds 38, 39, 43, 44, 55, 58, 59, and 60 still have efficacy, but the efficacy is weaker than 96h; compounds 31, 33, 35, 37, 40, 49-54 have the second best efficacy; followed by compounds 29 and 30, which have weaker efficacy; compared with the PBS control group, compounds 28, 32, 34, 36, 41, 42, 45-48, 56-57, 61 and Semaglutide have no significant difference and no efficacy, which is consistent with the no efficacy results of 72h and 96h.

[0260] (6) 144h results. Figure 15-16 It can be seen that compounds 38, 39, 43, 44, 55, and 58 still have efficacy, but the efficacy is weaker than 120h; compounds 49, 50, 51, 52, 53, 54, and 59 have the second best efficacy, followed by compounds 31, 33, 35, 37, and 40, which have weaker efficacy; compared with the PBS control group, compounds 28, 29, 30, 32, 34, 36, 41, 42, 45-48, 56-57, 60, 61 and Semaglutide have no significant differences and have lost their efficacy.

[0261] (7) 168h results. Figure 17-18 It can be seen that compared with the PBS control group, compounds 38, 39, 43, 44, 55, and 58 still have weak efficacy; 49, 50, 51, 52, 53, 54, and 59 have the second best effect; and the remaining compounds have lost their efficacy.

[0262] Conclusion: Analysis of the above results shows that although Semaglutide and each compound have good effects in improving glucose tolerance, the duration of drug effect varies among the compounds, that is, in terms of long-term glucose lowering. Among them, compounds 38, 39, 43, 44, 55, 58, 49, 50, 51, 52, 53, 54, and 59 have obvious advantages in both drug effect and drug effect duration (long-term effect), and are far superior to Semaglutide.

[0263] Example 4: Therapeutic effects of compounds 31, 33, 35, 38, 39, 43, 44, 49, 58, 60 and Semaglutide on BKS-db diabetic mice

[0264] Based on the results of the OGTT experiment, we further studied the efficacy of compounds 31, 33, 35, 38, 39, 43, 44, 49, 58, and 60 in the BKS-db diabetic mouse model and examined the effects of the compounds on body weight and blood sugar.

[0265] Experimental methods: This experiment used 8-week-old BKS-db diabetic mice with the Lepr KO / KO genotype (purchased from Guangdong Yaokang Biotechnology Co., Ltd.). Blood glucose and body weight were measured, and the mice were randomly divided into compound groups, positive control group (Semaglutide) and model control group (PBS) with 6 mice in each group. Each group of mice was subcutaneously injected with compounds 31, 33, 35, 38, 39, 43, 44, 49, 58, 60, and Semaglutide (injection solution purchased from Guangzhou Tonghui Pharmaceutical Co., Ltd.) at a dose of 120 μg / kg, once every other day. The control group was injected with an equal volume of normal saline. The experimental period was 4 weeks. After each administration, the mice were fasted for 6 hours every other day, and the blood glucose and body weight of the mice were tested and the data were recorded. The data were processed using the software GraphPadPrism. The experimental results are shown in the figure. Figure 19-20 , where * indicates p<0.05; ** indicates p<0.01; *** indicates p<0.001; **** indicates p<0.0001.

[0266] Result analysis:

[0267] (1) Statistical analysis of mouse weight monitoring data is shown in Fig.19 The results showed that compared with the model control group (PBS), compounds 31, 33, 35, 38, 39, 43, 44, 49, 58, and 60 could effectively reduce the body weight of mice, and the weight reduction effect was better than Semaglutide.

[0268] (2) Statistical analysis of fasting blood glucose monitoring data of mice is shown in Fig. 20The results showed that compared with the model control group (PBS), compounds 31, 33, 35, 38, 39, 43, 44, 49, 58, 60 and Semaglutide could significantly reduce the fasting blood glucose level of BKS-db diabetic mice, indicating that these compounds have significant hypoglycemic effects. In the first week, blood glucose had dropped to normal levels, and thereafter, blood glucose in the compound 31, 33, 35, 38, 39, 43, 44, 49, 58, 60 groups was relatively stable and the effect was better than Semaglutide.

[0269] Example 5: Pharmacodynamics validation test of lixisenatide and its modified peptides

[0270] Compound 62 synthesized in the present invention is the marketed GLP-1 receptor agonist polypeptide drug lixisenatide, and compound 63 is the side chain Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 The compound obtained by modifying the 12th Lys of lixisenatide with H, compound 64 is the side chain Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 The compounds obtained by modifying Lys 26 of lixisenatide with H. The pharmacodynamic effects of compound 62, compound 63 and compound 64 on oral glucose tolerance (OGTT) in mice were studied.

[0271] Experimental method: Male C57BL / 6J mice of about 8 weeks old (purchased from Guangdong Sijia Jingda Biotechnology Co., Ltd.) were raised for one week to adapt to the environment and randomly divided into groups according to blood sugar, with 8 mice in each group. Each polypeptide compound was administered subcutaneously at a dose of 80ug / kg, and the control group was given an equal volume of PBS. Fasting for 16 hours on the evening of the administration, 24 hours after administration, glucose was gavaged at a dose of 2g / kg, and blood sugar levels were detected at t=0min, t=15min, t=30min, t=60min, t=90min and t=120min. The software GraphPadPrism was used to process the data, draw a time-blood sugar curve, and calculate the area under the blood sugar curve AUC. One-way ANOVA analysis was performed with the PBS control group without administration to calculate the significant difference.

[0272] Result analysis: According to the OGTT result data statistics chart ( Figure 21-22) showed that: 24h after administration, compared with the vehicle (PBS), compound 62 (lixisenatide) could significantly reduce the AUC, indicating that lixisenatide has significant glucose tolerance and can effectively lower blood sugar; while compounds 63 and 64 after side chain modification of lixisenatide had no significant effect on AUC, indicating that peptide modification relying solely on side chain modification cannot improve glucose tolerance and cannot effectively lower blood sugar.

[0273] Conclusion: Adding the side chain involved in this application to lixisenatide not only fails to prolong its hypoglycemic effect, but makes the original peptide lose its hypoglycemic effect. Therefore, it can be explained that the side chain connection of the polypeptide compound involved in the invention of this application is not generally applicable to other peptides, but has its own creativity.

[0274] Example 6: Pharmacokinetic study of compound 33, compound 35, compound 38, compound 39, compound 43, compound 44, compound 58 and compound 61 in SD rats

[0275] SD rats (SPF grade, source: Sbefor (Beijing) Biotechnology Co., Ltd., experimental animal production license number: SCXK (Beijing) 2019-0010, weight: 180-200g, age: 6-8 weeks) were raised for one week to adapt to the environment. The general condition of the animals was checked during the adaptation period. Unqualified animals were not included in this experiment. The feeding environment control system used WINCC7.3 EMS series computer room environment monitoring system, and the feed used SPF rat maintenance feed. 20 SD rats that were raised and qualified were randomly divided into groups according to their weight, with 4 rats in each group (half male and half female), for a total of 5 groups (animals were grouped and statistically analyzed using Stata 15 software).

[0276] In this example, the compound was administered subcutaneously to the skin of the back of the neck at a dose of 0.15 mg / kg or 0.2 mg / kg, a dosing volume of 2 mL / kg, a dosing concentration of 0.075 mg / mL or 0.1 mg / mL, and a solvent of PBS.

[0277] Blood samples were collected from rats in the single subcutaneous administration group before administration (0 h) and 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h, 168 h, and 192 h after administration through the jugular vein. Approximately 0.2 mL of whole blood was collected at each time point and then lysed in EDTA-K 2In the anticoagulation tube, centrifuge at 4°C and 1800g centrifugal force for 10 minutes within 1 hour, take the supernatant, and transfer the separated plasma to a -80°C refrigerator for storage. UPLC-MS / MS was used to establish the concentration analysis method of the compound in SD rat plasma and determine the drug concentration of the compound in plasma. WinNonlin 8.1 software was used for data processing to calculate the pharmacokinetic parameters. The experimental results are shown in Table 2.

[0278] The experimental results show that compared with compound 61, after a single subcutaneous administration of compound 33, compound 35, compound 38, compound 39, compound 43, compound 44, and compound 58, the compounds were absorbed more slowly in rats, with a peak time T max The half-life was about 24 h, which was significantly higher than the 8 h of compound 61. 1 / 2 The average half-life was 12.2 h, 17.8 h, 18.8 h, 22.3 h, 25.5 h, 21.7 h, and 20.4 h, respectively, which was significantly higher than the 10.8 h of compound 61, and the half-life of compound 39 was more than 22 h. This shows that the long-acting polypeptide compound of the present invention has a longer half-life.

[0279] Table 2 Results of pharmacokinetic experiments in SD rats and corresponding dosages of the compounds

[0280] Compound <![CDATA[T 1 / 2 (h)]]> <![CDATA[T max (h)]]> Dosage mg / kg 33 12.2 24 0.2 35 17.8 24 0.2 38 21.8 24 0.2 39 22.3 24 0.2 43 20..5 24 0.2 44 21.7 24 0.2 58 20.4 24 0.2 61 10.8 8 0.15

[0281] The above are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art who is familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A novel long-acting polypeptide compound, characterized in that: The long-acting polypeptide compound is selected from any one of the following compounds: Compound 28: <h2 style=";text-align:left;direction:ltr">Y-Aib-QGTFTSDYSI-Aib-LDKIAQK(GGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGG PSSG-Aib-PPPS-NH2; Compound 29: <h2 style=";text-align:left;direction:ltr">Y-Aib-QGTFTSDYSI-Aib-LDKIAQK(GGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 30: <h2 style=";text-align:left;direction:ltr">Y-Aib-EGTFTSDYSI-Aib-LDKIAQK(GGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 31: <h2 style=";text-align:left;direction:ltr">Y-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLIAGGPSSGAPPPS-NH2; Compound 33: <h2 style=";text-align:left;direction:ltr">Y-Aib-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLIAGGPSSGAPPPS-NH2; Compound 35: <h2 style=";text-align:left;direction:ltr">Y-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLIAGGPSSGAPPPS-NH2; Compound 37: <h2 style=";text-align:left;direction:ltr">Y-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 38: <h2 style=";text-align:left;direction:ltr">Y-Aib-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 39: <h2 style=";text-align:left;direction:ltr">Y-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 40: <h2 style=";text-align:left;direction:ltr">Y-Iva-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 41: <h2 style=";text-align:left;direction:ltr">Y-Cba-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 42: <h2 style=";text-align:left;direction:ltr">Y-Cba-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 43: <h2 style=";text-align:left;direction:ltr">Y-Aib-EGTFTSDYSI-Cba-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 44: <h2 style=";text-align:left;direction:ltr">Y-Iva-EGTFTSDYSI-Cba-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-NH2; Compound 45: H-Aib-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLAGGPSSGAPPPS-OH; Compound 46: <h2 style=";text-align:left;direction:ltr">H-Iva-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLAGGPSSGAPPPS-OH; Compound 47: <h2 style=";text-align:left;direction:ltr">H-Iva-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLAGGPSSGAPPPS-OH; Compound 48: <h2 style=";text-align:left;direction:ltr">H-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLAGGPSSGAPPPS-OH; Compound 49: H-Iva-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-OH; Compound 50: H-Iva-EGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-OH; Compound 51: H-Cba-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLIAGGPSSGAPPPS-OH; Compound 52: H-Cba-QGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-OH; Compound 53: H-Aib-QGTFTSDYSI-Iva-LDKIAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLIA GGPSSG-Aib-PPPS-OH; Compound 54: <h2 style=";text-align:left;direction:ltr">H-(D-Ser)-QGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLLDGGPSSGAPPPS-OH; Compound 55: Y-Iva-EGTFTS (D) YSI-Iva-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLIAGGPSSGAPPPS-NH2, wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond; Compound 56: Y-Aib-EGTFTS (D) YSI-Iva-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-NH2, wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond; Compound 57: Y-Iva-EGTFTS (D) YSI-Iva-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-NH2, wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond; Compound 58: H-Aib-QGTFTS (D) YSI-Aib-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond; Compound 59: H-Iva-EGTFTS (D) YSI-Aib-LD (K) IAQK(GGSGSGSG-γGlu-CO(CH2) 18 CO2H)AFVQWLLDGGPSSGAPPPS-OH, wherein the 9th amino acid D is connected to the 16th amino acid K through an amide bond; Compound 60: <h2 style=";text-align:left;direction:ltr">Y-Aib-EGTFTSDYSI-Aib-LDKIAQK(GGSGSGSG-γGlu-CO(CH2)<h2 style=";text-align:left;direction:ltr"> 18 <h2 style=";text-align:left;direction:ltr"> CO2H)AFVQWLIAGGPSSGAPPPS-NH2。 2. The long-acting polypeptide compound according to claim 1, characterized in that: The long-acting polypeptide compound is a pharmaceutically acceptable salt.

3. A composition, characterized in that The composition comprises the long-acting polypeptide compound according to claim 1 or 2 and a pharmaceutically acceptable carrier or excipient.

4. Use of the long-acting polypeptide compound according to claim 1 or 2 or the composition according to claim 3 in the preparation of a medicament for preventing or treating diabetes; Alternatively, the long-acting polypeptide compound or the composition is used in the preparation of a drug for preventing or treating obesity.

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