GLP-1 polypeptide compound and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a GLP-1 polypeptide compound and application thereof. In particular to a GLP-1 polypeptide compound or pharmaceutically acceptable salt, amide or ester thereof. The amino acid sequence of the GLP-1 polypeptide compound is shown as a formula (I) shown in SEQ ID NO: 13: HX2EGTFTSDVSYLEGQAAX20NFVX24X25LIARAEX32 (I); wherein X2 is aminoisobutyric acid; x20 is K with a modified side chain and is COOH-(CH2) a-CO-(gamma Glu) b-(NH-((CH2) 2-O) 2-CO) 2-K, a is any integer selected from 16-20, and b is 1 or 2; x24 is an amino acid selected from A, Q, or R; x25 is an amino acid selected from Q or R; x32 is A-R1 or-R1, R1 is a modification of a carboxyl group at the C terminal of the sequence, and the modification is NH2 (amidation) or does not exist.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a GLP-1 polypeptide compound and application thereof. BACKGROUND

[0002] In the past three decades, the prevalence of diabetes mellitus in China has increased significantly. Type 2 diabetes (T2D) is the most common form of diabetes, accounting for more than 90% of all diabetic patients. T2D is caused by insulin resistance and non-autoimmune beta cell insulin secretion deficiency caused by metabolic syndrome. At present, the care methods for T2D include lifestyle adjustment and blood glucose control drugs, however, the blood glucose level control method for T2D patients is still insufficient, and new drugs need to be further developed.

[0003] Current T2D drugs include metformin, sulfonylureas, glucagon-like peptide-1 (GLP-1) receptor agonists, insulin, etc. Metformin is a first-line drug for the treatment of diabetes, but 30% of patients in clinical practice have poor efficacy or adverse reactions that cannot be tolerated, and in severe cases, acidosis may even occur. And with the progression of the disease, metformin alone will be increasingly difficult to control blood glucose. If sulfonylurea drugs are not used properly, they can easily cause hypoglycemia and weight gain, and individual patients may experience skin allergic reactions, leukopenia, etc. And affected by factors such as dose size, injection site, injection route, individual differences, or not eating after injection, if insulin is used slightly carelessly, serious hypoglycemia side effects may occur. Although new hypoglycemic mechanism drugs such as SGLT2i have been launched in recent years, they do not increase the risk of hypoglycemia when used alone, but they also expose the shortcomings of increasing the risk of urinary / genital system infections.

[0004] Glucagon-like peptide-1 (GLP-1) is an intestinal insulin, GLP-1 and its analog GLP-1 have glucose concentration-dependent hypoglycemic effect, which avoids the risk of hypoglycemia often present in diabetes treatment. Metabolism in the body is a complex and interdependent process, and other metabolic abnormalities can also significantly increase the risk, rate of progression, and harm of T2D complications. Current studies have shown that multiple GLP-1 receptor agonists can not only significantly improve blood glucose control, but also improve lipid metabolism abnormalities, hypertension, and body weight, in addition, they can also benefit the cardiovascular and kidney systems, and have broad development prospects.

[0005] The intellectual property rights of current GLP-1 analog innovative drugs are mainly concentrated in multinational pharmaceutical companies such as Novo Nordisk and Lilly. Novo Nordisk's semaglutide, Lilly's dulaglutide, and tirzepatide, which has not yet entered the Chinese market, are all long-acting preparations that are injected once a week.

[0006] Therefore, it is urgent to develop more safe and widely used long-acting GLP-1 analogs to fill the serious shortage of self-developed blood sugar control drugs in China. SUMMARY

[0007] In order to solve the problems existing in the prior art, the purpose of the present disclosure is to provide a GLP-1 polypeptide compound and application thereof.

[0008] To solve the above technical problems, the present disclosure adopts the following technical solutions:

[0009] In one aspect, the present disclosure provides a GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, having an amino acid sequence as shown in SEQ ID NO: 13 of formula (I):

[0010] HX2EGTFTSDVSSYLEGQAAX 20 NFVX 24 X 25 LIARAEX 32 (I);

[0011] wherein X2 is aminoisobutyric acid; X 20 is a side chain modified K, which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, wherein a is any integer selected from 16-20, and b is 1; X 24 is an amino acid selected from A, Q, or R; X 25 is an amino acid selected from Q or R; X 32 is A-R 1 or -R 1 , R 1 is a modification of the carboxyl group at the C-terminal end of the sequence, wherein the modification is NH2 or absent.

[0012] In another aspect, the present disclosure provides a pharmaceutical composition comprising the aforementioned GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, and a pharmaceutically acceptable carrier, adjuvant or excipient;

[0013] Preferably, the dosage form of the pharmaceutical composition is selected from any one of tablets, capsules, elixirs, syrups, lozenges, inhalants, sprays, injections, films, patches, powders, granules, blocks, emulsions, suppositories or compound preparations.

[0014] In another aspect, the present disclosure provides a use of the aforementioned GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing or treating diabetes, obesity, hypertension, dyslipidemia, atherosclerosis, coronary heart disease and other cardiovascular diseases, stroke, Alzheimer's disease, non-alcoholic fatty liver disease and other metabolic syndromes.

[0015] In another aspect, the present disclosure provides a method for preparing the aforementioned GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, wherein the method is by chemical synthesis.

[0016] The GLP-1 polypeptide compound of the present disclosure has a non-proteinogenic amino acid residue at the 2nd position in the sequence, which reduces its sensitivity to dipeptidyl peptidase IV (DPP-IV) and increases its proteolytic stability. The lysine residue at the 20th position has a fatty acid chain connected by a linker, which further prolongs the half-life of the relevant GLP-1 analogue in the blood by binding to albumin, improves the pharmacokinetics of the peptide, and supports the pharmacokinetic (PK) characteristics of once-weekly dosing in humans. At the same time, the hypoglycemic and weight loss effects of the polypeptide compound Dia2-004 of the present disclosure are greater than those of semaglutide. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 The chromatographic results of Dia2-001 are shown.

[0018] Figure 2 The mass spectrometric results of Dia2-001 are shown.

[0019] Figure 3 The chromatographic results of Dia2-002 are shown.

[0020] Figure 4 The mass spectrometric results of Dia2-002 are shown.

[0021] Figure 5 The chromatographic results of Dia2-003 are shown.

[0022] Figure 6 The mass spectrometric results of Dia2-003 are shown.

[0023] Figure 7 The chromatographic results of Dia2-004 are shown.

[0024] Figure 8 The mass spectrometric results of Dia2-004 are shown.

[0025] Figure 9 The chromatographic results of Dia2-005 are shown.

[0026] Figure 10 Mass spectrometry results of Dia2-005 are shown.

[0027] Figure 11 Chromatography results of Dia2-006 are shown.

[0028] Figure 12 Mass spectrometry results of Dia2-006 are shown.

[0029] Figure 13 Chromatography results of Dia2-009 are shown.

[0030] Figure 14 Mass spectrometry results of Dia2-009 are shown.

[0031] Figure 15 Chromatography results of Dia2-010 are shown.

[0032] Figure 16 Mass spectrometry results of Dia2-010 are shown.

[0033] Figure 17 The GLP-1R agonistic potency concentration dependent fitting curves of semaglutide and Dia2-001-010 are shown.

[0034] Figure 18 The effect of GLP-1R agonistic polypeptides on blood glucose in mice in a glucose tolerance test after a single subcutaneous dose of 3 hours is shown.

[0035] Figure 19 The results of the effect of polypeptide compounds on blood glucose in db / db mice within 0-48h in Example 5 are shown in the graph.

[0036] Figure 20 The results of the effect of polypeptide compounds on body weight in db / db mice within 0-48h in Example 5 are shown in the graph. DETAILED DESCRIPTION

[0037] I. DEFINITIONS

[0038] In the present disclosure, the scientific and technical terms used herein have the meanings commonly understood by a person of ordinary skill in the art, unless otherwise indicated. Also, the terms and procedures employed in protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, chemistry, and related scientific disciplines are those commonly used by persons skilled in the respective fields. Also, for better understanding of the present disclosure, the definitions and explanations of relevant terms are provided below.

[0039] The terms "comprise" and "have", and any variations thereof, are intended to cover a non-exclusive inclusion. For example, a process, method, article, or apparatus that comprises a list of steps or modules does not necessarily require the listed steps or modules to be present, but can optionally be present, or can further include additional steps or modules not listed. In the present disclosure, "a plurality of" means two or more. "And / or", describes the association relationship of associated objects, which means that there can be three relationships, for example, A and / or B can represent: A exists alone, A and B exist together, B exists alone. The character " / " generally represents that the associated objects before and after are a "or" relationship.

[0040] The term "or its combination" as used herein refers to all permutations and combinations of the terms listed before the term. For example, "A, B, C, or a combination thereof" is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, also BA, CA, CB, ACB, CBA, BCA, BAC, or CAB. Continuing with this example, explicitly included are combinations that have repetitions of one or more items or terms, such as BB, AAA, AAB, BBC, AAABCCCC, CBBAAA, CABABB, and the like. The skilled person will understand that there is generally no limitation on the number of items or terms in any combination, unless otherwise explicitly apparent from the context.

[0041] In terms of polypeptide sequences, the phrase "substantially identical" can be understood to exhibit at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a reference polypeptide sequence. In terms of nucleic acid sequences, the term can be understood as a nucleotide sequence that exhibits at least greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a reference nucleic acid sequence.

[0042] In the description herein, reference is made to "some embodiments", "some implementations" or "some aspects", which describe a subset of all possible embodiments, but it can be understood that "some embodiments" can be the same subset or different subsets of all possible embodiments, and can be combined with each other without conflict.

[0043] As used herein, the term "amino acid" refers to a molecule that contains both an amino and a carboxyl functional group, the amino and carboxyl of an alpha-amino acid are attached to the same carbon atom (the alpha carbon). The alpha carbon can additionally have 1-2 organic substituents. Amino acids include L and D isomers and racemic mixtures. Unless otherwise specified, the amino acid residues in the polypeptide sequences of the disclosure are the L isomers, i.e., L-amino acids, D-amino acids are indicated by a lower case "d" before the amino acid name or abbreviation, e.g., dK.

[0044] The amino acid sequences of the disclosure contain the conventional one-letter or three-letter codes for naturally occurring amino acids, as well as the universally accepted three-letter codes for other amino acids, such as Iva (Isovaline); Cba (1-Aminocyclobutanecarboxylic acid); Tic (1,2,3,4-Tetrahydroisoquinoline-3-carboxylic acid); Aib (α-Aminoisobutyric acid); or GABA (γ-Aminobutyric acid). Abbreviation codes for commonly used molecular structures include:

[0045] hGlu is homoglutamic acid;

[0046] a-hGlu is the L isomer of -HNCH(CO-)CH2CH2CH2COOH;

[0047] δ-hGlu is the L isomer of -HNCH(COOH)CH2CH2CH2CO-;

[0048] a-Glu is the L isomer of -HNCH(CO-)CH2CH2COOH;

[0049] γ-Glu or gGlu is the L isomer of -HNCH(COOH)CH2CH2CO-;

[0050] a-Asp is the L isomer of -HNCH(CO-)CH2COOH;

[0051] β-Asp is the L isomer of -HNCH(COOH)CH2CO-;

[0052] β-Ala is -HN-CH2-CH2-COOH;

[0053] PEG2 is 2-(2-(2-Aminoethoxy)ethoxy)acetic acid (CAS No. 134978-97-5).

[0054] The amino acid composition of the polypeptides in the present disclosure can be altered without substantially affecting their biological activity. For example, a polypeptide sequence can contain one or more conservative amino acid substitutions. A conservative amino acid substitution is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. The literature is replete with classification schemes of amino acid residues based on the properties of their side chains. Basic side chain-containing amino acid residues include lysine, arginine, histidine; acidic side chain- and amide side chain-containing amino acid residues include aspartic acid, glutamic acid, asparagine, glutamine; small aliphatic, nonpolar or weakly polar side chain-containing amino acid residues include glycine, alanine, threonine, serine, proline; large aliphatic, nonpolar side chain-containing amino acid residues include leucine, isoleucine, valine; aromatic amino acid residues include phenylalanine, tryptophan, tyrosine; sulfur-containing side chain-containing amino acid residues include cysteine, methionine.

[0055] As used herein, the term "treatment" includes inhibiting, slowing, stopping, or reversing the progression or severity of an existing symptom or condition. Thus, treatment includes prevention, therapy, and / or cure. Prevention refers to preventing an underlying disease and / or preventing the worsening of symptoms or development of the disease. As used herein, "therapeutic effect" means an effect resulting from the treatment of an individual that alters, usually improves, or ameliorates the symptoms of a disease or disease condition, or cures the disease or disease condition. As used herein, "therapeutically effective amount" or "therapeutically effective dose" means an amount of a substance, compound, material, or composition comprising a compound that, after administration to a subject, is at least sufficient to produce a therapeutic effect. Thus, it is an amount necessary to prevent, cure, ameliorate, retard, or partially retard the symptoms of a disease or disorder. As used herein, "prophylactically effective amount" or "prophylactically effective dose" means an amount of a substance, compound, material, or composition comprising a compound that, when administered to a subject, will have the intended prophylactic effect, e.g., prevent or delay the onset of a disease or symptoms, reduce the likelihood of the onset of a disease or symptoms. A fully prophylactically effective dose does not necessarily occur with the administration of one dose, and can only occur after administration of a series of doses. Thus, a prophylactically effective amount can be administered in one or more administrations.

[0056] As used herein, the term "patient" refers to a mammal, such as a human.

[0057] Certain compounds of the present disclosure are typically effective over a wide dosage range. For example, dosages for once-weekly administration can range from about 0.05 to about 30 mg per person per week. Certain compounds of the present disclosure can be administered daily. In addition, certain compounds of the present disclosure can be administered once-weekly.

[0058] It will be appreciated that the therapeutic agents according to the described embodiments will be administered with a suitable pharmaceutically acceptable carrier, excipient, and other agents that are incorporated to provide improved transfer, delivery, tolerance, etc. A large number of suitable formulations are available in the pharmacopeias known to all pharmaceutical chemists. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid-containing (cationic or anionic) carriers (e.g., Lipofectin™), DNA conjugates, anhydrous absorbents, oil-in-water and water-in-oil emulsions, emulsions in polyethylene glycol (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing polyethylene glycol. Any of the aforementioned mixtures can be suitable for use in the treatment or therapy according to the present disclosure, provided that the active ingredients in the formulation are not inactivated by the formulation and the formulation is physiologically compatible and tolerable to the route of administration.

[0059] As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, which is standard reference text in the field, incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solutions, dextrose solutions, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils can also be used. Such media and agents for pharmaceutically active substances are well known in the art.

[0060] The formulations to be used for in vivo clinical administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.

[0061] In addition, the abbreviations used herein and their meanings are as follows:

[0062] T2D: Type 2 diabetes mellitus;

[0063] GLP-1: Glucagon-like peptide-1;

[0064] DPP-IV: Dipeptidyl peptidase IV;

[0065] Aib: Amino isobutyric acid;

[0066] tBu: tert-Butyl;

[0067] Fmoc: 9-Fluorenylmethyloxycarbonyl protecting group;

[0068] Alloc: Allyloxycarbonyl.

[0069] II. DETAILED DESCRIPTION

[0070] In one aspect, the present disclosure provides a GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, having an amino acid sequence of Formula (I) as shown in SEQ ID NO: 13:

[0071] HX2EGTFTSDVSSYLEGQAAX 20 NFVX 24 X 25 LIARAEX 32 (I);

[0072] wherein X2is aminoisobutyric acid; X 20 is a K with a modified side chain, which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-O)2-CO)2-K, wherein a is any integer selected from 16-20, b is 1; X 24 is an amino acid selected from A, Q, or R; X 25 is an amino acid selected from Q or R; X 32 is A-R 1 or -R 1 , R 1 is a modification of the C-terminal group of the sequence, wherein the modification is NH2or absent.

[0073] In some embodiments, X 20 is a K with a modified side chain, and the modified compound is attached to the epsilon-amino group of the K side chain.

[0074] In some embodiments, a is 16 or 18.

[0075] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof is selected from any one of the amino acid sequences shown in SEQ ID NOs: 1-12:

[0076] (1) SEQ ID NO. 1:

[0077] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-yGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAE;

[0078] (2) SEQ ID NO. 2:

[0079] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-yGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAE;

[0080] (3) SEQ ID NO. 3:

[0081] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE;

[0082] (4) SEQ ID NO. 4:

[0083] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE;

[0084] (5) SEQ ID NO. 5:

[0085] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE;

[0086] (6) SEQ ID NO. 6:

[0087] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE;

[0088] (7) SEQ ID NO. 7:

[0089] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE;

[0090] (8) SEQ ID NO. 8:

[0091] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE;

[0092] (9) SEQ ID NO. 9:

[0093] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA;

[0094] (10) SEQ ID NO. 10:

[0095] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA;

[0096] (11) SEQ ID NO. 11:

[0097] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAEA;

[0098] (12) SEQ ID NO. 12:

[0099] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAEA.

[0100] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K that is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 18, b is 1; X 24 is Q; X 25 is R; X 32 is -R 1 , R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAE.

[0101] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X20 K, which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 18, b is 1; X 24 Q; X 25 R; X 32 -R 1 R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVQRLIARAE.

[0102] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 K, which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X 24 R; X 25 Q; X 32 -R 1 R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVQRLIARAE.

[0103] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 K, which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 18, b is 1; X 24 R; X 25 Q; X 32 -R 1 R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVQRLIARAE.

[0104] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, wherein a is 18, b is 1; X 24 is A; X 25 is R; X 32 is -R 1 R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K}NFVARLIARAE.

[0105] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, wherein a is 18, b is 1; X 24 is A; X 25 is R; X 32 is -R 1 R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K}NFVARLIARAE.

[0106] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, wherein a is 16, b is 1; X 24 is Q; X 25 is R; X 32 is A; R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2)16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAEA.

[0107] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is K modified at the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X 24 is Q; X 25 is R; X 32 is A; R 1 is absent, and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAEA.

[0108] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is K modified at the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X 24 is R; X 25 is Q; X 32 is A; R 1 is absent, and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA.

[0109] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is K modified at the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 18, b is 1; X 24 is R; X 25 is Q; X 32 is A; R 1is absent and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRQLIARAEA.

[0110] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a K modified in the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, wherein a is 18, b is 1; X 24 is A; X 25 is R; X 32 is A; R 1 is absent and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVARLIARAEA.

[0111] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a K modified in the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, wherein a is 18, b is 1; X 24 is A; X 25 is R; X 32 is A; R 1 is absent and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVARLIARAEA.

[0112] In some preferred embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof is selected from any one of the following amino acid sequences:

[0113] (1) SEQ ID NO. 2:

[0114] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18-CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVQRLIARAE;

[0115] (2) SEQ ID NO. 3:

[0116] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRQLIARAE;

[0117] (3) SEQ ID NO. 4:

[0118] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRQLIARAE;

[0119] (4) SEQ ID NO. 5:

[0120] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVARLIARAE;

[0121] (5) SEQ ID NO. 6:

[0122] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVARLIARAE.

[0123] (6) SEQ ID NO. 10:

[0124] HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRQLIARAEA;

[0125] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2 is aminoisobutyric acid, X 20 is a side chain modified K that is COOH-(CH2) a -CO-(γGlu) b-(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 24 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 25 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 32 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 1 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 1 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2)

[0126] -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 20 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) a -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) b -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 24 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 25 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 32 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 1 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 1 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2)

[0127] -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 20 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) a -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) b -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 24 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 25 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 32 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 1 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2)

[0128] -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X is A; X is Q; R is -R; R is absent; and the amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2)20 K, which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, where a is 18, b is 1; X 24 A; X 25 Q; X 32 A; R 1 is absent, and its amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K} NFVAQLIARAEA.

[0129] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 K, which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, where a is 16, b is 1; X 24 Q; X 25 Q; X 32 R 1 R 1 is absent, and its amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K} NFVQQLIARAE.

[0130] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 K, which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, where a is 18, b is 1; X 24 Q; X 25 Q; X 32 -R 1 R 1 is absent, and its amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K} NFVQQLIARAE.

[0131] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, wherein a is 16 and b is 1; X 24 is Q; X 25 is Q; X 32 is A, R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K}NFVQQLIARAEA.

[0132] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, wherein a is 18 and b is 1; X 24 is Q; X 25 is Q; X 32 is A, R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-yGlu-(NH-((CH2)2-0)2-CO)2-K}NFVQQLIARAEA.

[0133] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(yGlu) b -(NH-((CH2)2-0)2-CO)2-K, wherein a is 16 and b is 1; X 24 is R; X 25 is R; X 32 is -R 1 , R 1 is absent; and has the amino acid sequence HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16-CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRRLIARAE.

[0134] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is K modified in the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 18, b is 1; X 24 is R; X 25 is R; X 32 is -R 1 , R 1 is absent, and has the amino acid sequence of HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRRLIARAE.

[0135] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is K modified in the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 16, b is 1; X 24 is R; X 25 is R; X 32 is A; R 1 is absent, and has the amino acid sequence of HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K} NFVRRLIARAEA.

[0136] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, wherein X2is aminoisobutyric acid, X 20 is K modified in the side chain which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is 18, b is 1; X 24 is R; X 25 is R; X 32 is A; R 1absent, and its amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGl u-(NH-((CH2)2-O)2-CO)2-K}NFVRRLIARAEA.

[0137] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, wherein a is 18, b is 1; X 24 is R; X 25 is R; X 32 is A; R 1 absent, and its amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-(γGlu)2-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA.

[0138] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, wherein X2is aminoisobutyric acid, X 20 is a side chain modified K which is COOH-(CH2) a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, wherein a is 18, b is 1; X 24 is R; X 25 is R; X 32 is A; R 1 absent, and its amino acid sequence is HAibEGTFTSDVSSYLEGQAA{COOH-(CH2)18-CO-(γGlu)2-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA.

[0139] In some embodiments, the side chain (modifying the 20th amino acid) of the polypeptide compound in the present disclosure, which mainly functions to bind with serum albumin, prolongs the retention time of the polypeptide compound in the blood, thereby prolonging the half-life of the polypeptide in vivo, and provides the potential to generate a long-acting compound. In some embodiments, the present disclosure further modifies the structure of the aforementioned polypeptide compound, i.e., adding one γGlu to the γGlu position in the side chain K in the aforementioned polypeptide compound, so that the side chain COOH-(CH2) a-CO-(yGlu) b -(NH-((CH2)2-O)2-CO)2-K, b is 2, can also obtain the effect of binding with serum albumin.

[0140] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, wherein the salt is a salt of the GLP-1 polypeptide compound with one of the following compounds: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric 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, cyclopentane propionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, 2-naphthalenesulfonic acid, naphthalene disulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid, or thiocyanic acid.

[0141] In some embodiments, the salt is a salt of the GLP-1 polypeptide compound with one of the following compounds: trifluoroacetic acid or acetic acid.

[0142] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof is modified at its N- or C-terminus with a helper group.

[0143] In some embodiments, the GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof is modified at its N- or C-terminus with a helper group.

[0144] In some embodiments, the dosage form of the pharmaceutical composition is selected from any one of a tablet, a capsule, an elixir, a syrup, a lozenge, an inhaler, a spray, an injection, a film, a patch, a powder, a granule, a block, an emulsion, a suppository, or a compound preparation.

[0145] In some embodiments, the dosage form of the pharmaceutical composition is selected from any one of a tablet, a capsule, an elixir, a syrup, a lozenge, an inhaler, a spray, an injection, a film, a patch, a powder, a granule, a block, an emulsion, a suppository, or a compound preparation.

[0146] In another aspect, the present disclosure provides a method of preparing the aforementioned GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof, wherein the method of preparation is by chemical synthesis.

[0147] In some embodiments, the method comprises the following steps:

[0148] (1) Resin swelling. Using Fmoc / t-Bu strategy for solid phase peptide synthesis, start from RAPP AM-Rink amide resin, weigh the corresponding amount of resin, pour into the polypeptide solid phase synthesis tube, according to the ratio of 1 g to 10 mL (solvent just covers the resin), swell the resin with dichloromethane, and the swelling time is about 30 min to 2 h. After the resin is completely swollen, the solvent is removed; according to the ratio of 1 g to 10 mL, add DMF to wash the resin for 1 min, then remove the solvent under vacuum, add 20% piperidine DMF solution, remove the Fmoc protecting group, and the removal time is 5 min + 7 min. After the removal is completed, add DMF to wash three times, each for 1 min, and the indene test resin should be purple;

[0149] (2) Polypeptide sequence extension. According to the corresponding amount ratio (molar number of resin active groups: molar number of materials), add the reaction raw materials, AA (amino acid, 3 equivalents), HOAt (3 equivalents), condensing agent (3 equivalents), DIPEA (9 equivalents), solvent is DMF, nitrogen protection, reaction under room temperature for 1-2 h. Remove the solvent under vacuum, add DMF to wash three times, each for 1 min. Then take a small amount of resin for indene test, and the resin shows colorless, indicating that the condensation reaction is complete (indene test needs to use electric heating plate);

[0150] During the extension of the peptide chain, the Fmoc protecting group on the previous amino acid needs to be removed. Add 20% piperidine DMF solution (to cover the resin) to the reaction tube, and the removal time is 5 min + 7 min. After the removal is completed, remove the piperidine solution, and wash with DMF solution six times, each for 1 min. After Fmoc removal, the indene test resin should be purple (slightly different for different amino acids).

[0151] (3) Aib2 and His1 coupling. For Aib2, extended coupling (4 h each) is necessary to improve the quality of the crude peptide. Using DIC-HOBt scheme, introduce N-terminal residue in the form of Boc-His(tBu)-OH;

[0152] (4) Coupling and removal of Alloc from Lys20. Lys20 coupling (orthogonal protecting group) was performed using Fmoc-Lys(Alloc)-OH building block to allow site-specific attachment of the fatty acid moiety later in the synthesis. After completion of the main chain peptide resin coupling, 0.5 equivalents of tetrakis triphenylphosphine palladium, 20 equivalents of morpholine were added and the reaction was allowed to proceed for two hours in DCM, sampled for MS analysis and if correct, the side chain was coupled;

[0153] (5) Coupling of the side chain fragment. ([2-(2-amino-ethoxy)-ethoxy]- acetyl)2-γGlu(OtBu)-CO-(CH2) 18 -CO(OtBu) or ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γGlu(OtBu)-CO-(CH2) 16 -CO(OtBu) were purchased from Gilson Biochemical (Cas numbers: 1188328-37-1, 1118767-16-0, respectively). Four equivalents of the side chain fragment, four equivalents of PyBop, four equivalents of HOAT were added and the reaction was allowed to proceed for two hours in NMP, followed by K test;

[0154] (6) Cleavage. The resin was collapsed by addition of methanol. After completion of the collapse, the methanol was filtered off under reduced pressure and the resin was dried (the resin was powdery and free flowing after drying). The resin was then cleaved using classical cleavage solution to give the GLP1 series crude peptide;

[0155] (7) Purification. The crude peptide was purified to >95% purity by reverse phase HPLC chromatography on a C18 column using a water / acetonitrile (containing 0.05% v / v TFA) gradient (15-20% purification yield), and the appropriate fractions were combined and freeze-dried;

[0156] (8) Compound purity analysis and structure confirmation. The purity of Example 1 was checked by analytical reverse phase HPLC and the compound identity was confirmed by LC / MS.

[0157] The compound identity was confirmed by LC / MS. HPLC was performed using 0.1% trifluoroacetic acid in water (v / v) and acetonitrile as mobile phase, with a 30 minute elution program. LC / MS was performed using 0.1% formic acid in water and acetonitrile as mobile phase, with a 10 minute elution program.

[0158] In another aspect, the present disclosure provides a method for preventing or treating diabetes, obesity, hypertension, dyslipidemia, atherosclerosis, coronary heart disease and other cardiovascular diseases, stroke, Alzheimer's disease, non-alcoholic fatty liver disease and other metabolic syndromes, comprising administering a therapeutically effective amount of the aforementioned GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide or ester thereof or the aforementioned pharmaceutical composition.

[0159] The present disclosure will be described in more detail by referring to specific examples, which are merely for illustrative purposes and are not intended to limit the present disclosure. The reagents and biological materials used in the following examples are commercially available unless otherwise specified.

[0160] Examples

[0161] Example 1: Synthesis of polypeptide compounds

[0162] The method of polypeptide solid phase chemical synthesis is a well-developed methodology, which can be referred to the literature such as R.C. Sheppard, Solid Phase Peptide Synthesis. A Practical Approach, Oxford-IRL Press, New York, 1989.

[0163] The compound of the present disclosure is a straight-chain peptide. Each amino acid can be coupled step by step in the order of polypeptide sequence from C-terminal to N-terminal, so as to obtain the polypeptide backbone. The process is as follows: first, an amino acid with an amino group protected by a blocking group is covalently connected to a solid phase carrier, and the amino protecting group of the first amino acid is removed, so that the first amino acid is connected to the solid phase carrier. Then the carboxyl group of the second amino acid with an amino group blocked is activated and reacted with the amino group of the first amino acid connected to the solid phase carrier to form a peptide bond, so that a dipeptide with a protecting group is generated on the solid phase carrier. Repeat the above peptide bond formation reaction to extend the peptide chain from the C-terminal to the N-terminal until the desired peptide chain is generated. Finally, the protecting group is removed, and the covalent bond between the peptide chain and the solid phase carrier is hydrolyzed, to obtain the synthesized polypeptide. The specific experimental process is as follows:

[0164] (1) Resin swelling. Use Fmoc / t-Bu strategy for solid phase peptide synthesis, start from RAPP AM-Rink amide resin, weigh the corresponding amount of resin, pour into the polypeptide solid phase synthesis tube, according to the ratio of 1 g to 10 mL (solvent just covers the resin), use dichloromethane to swell the resin, the swelling time is about 30 min to 2 h. After the resin is completely swollen, the solvent is removed; according to the ratio of 1 g to 10 mL, add DMF to wash the resin for 1 min, then remove the solvent under vacuum, add 20% piperidine DMF solution to remove the Fmoc protecting group, the removal time is 5 min+7 min, after the removal is completed, add DMF to wash three times, each time for 1 min, the resin should present purple color.

[0165] (2) Polypeptide sequence elongation. Add the reactants according to the equivalent ratio (mole number of active groups of resin: mole number of material), AA (amino acid, 3 equivalents), HOAt (3 equivalents), condensing agent (3 equivalents), DIPEA (9 equivalents), and DMF as the solvent, under nitrogen protection, and react for 1-2 hours under stirring or shaking at room temperature. Remove the solvent under vacuum, and wash the resin with DMF three times, each time for 1 minute. Then, take a small amount of resin for indenyl test, and the resin appears colorless, indicating that the condensation reaction is complete (the indenyl test requires a hot plate).

[0166] During the elongation of the peptide chain, the Fmoc protecting group on the last amino acid needs to be removed. Add 20% piperidine DMF solution (to the resin) to the reaction tube, and remove the Fmoc protecting group for 5 minutes + 7 minutes. After the removal is complete, remove the piperidine solution, and wash the resin with DMF six times, each time for 1 minute. After the Fmoc protecting group is removed, the indenyl test resin should appear purple (slightly different for different amino acids).

[0167] (3) Coupling of Aib2 and His1. For Aib2, elongated coupling (4 hours each) is necessary to improve the quality of the crude peptide. Use the DIC-HOBt scheme to introduce the N-terminal residue in the form of Boc-His(tBu)-OH.

[0168] (4) Coupling of Lys20 and removal of Alloc. Use the Fmoc-Lys(Alloc)-OH building block to couple Lys20 (orthogonal protecting group) for later site-specific connection of the fatty acid moiety during the synthesis process. After the main chain peptide resin coupling is complete, add 0.5 equivalents of tetrakis triphenylphosphine palladium and 20 equivalents of morpholine, and react for two hours with DCM as the solvent. Take a sample for MS detection, and if the result is correct, couple the side chain.

[0169] (5) Attach the side chain fragment. ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γGlu(OtBu)-CO-(CH2) 18 -CO(OtBu) or ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-γGlu(OtBu)-CO-(CH2) 16 -CO(OtBu) purchased from Gilson Biochemical (CAS numbers: 1188328-37-1, 1118767-16-0). Add 4 equivalents of the side chain fragment, 4 equivalents of PyBop, and 4 equivalents of HOAT, and react for two hours with NMP as the solvent, followed by K test.

[0170] (6) Cleavage. Shrink the resin by adding methanol. After the shrinkage is complete, filter off the methanol under reduced pressure, and dry (the resin is powdery and can slide freely after drying). Then, use the classic cleavage solution to cut, and the GLP1 series of crude peptides can be obtained.

[0171] (7) Purification. The crude peptides were purified by reverse phase HPLC chromatography on C18 column with water / acetonitrile (containing 0.05% v / v TFA) gradient to >95% purity (15-20% purification yield), the appropriate fractions were combined and freeze-dried.

[0172] (8) Compound purity analysis and structure confirmation. The purity of Example 1 was checked by analytical reverse phase HPLC and the structure was confirmed by

[0173] Compound identity was confirmed by LC / MS. HPLC was performed with 0.1% trifluoroacetic acid in water (v / v) and acetonitrile as mobile phase, elution program for 30 minutes. LC / MS was performed with 0.1% formic acid in water and acetonitrile as mobile phase, elution for 10 minutes.

[0174] The synthesized polypeptide compounds are shown in Table 1 below.

[0175] Table 1. Amino acid sequence of polypeptide compounds and their salt forms

[0176]

[0177]

[0178] In addition, the applicant further synthesized the polypeptide compounds shown in Table 2.

[0179] Table 2. Amino acid sequence of polypeptide compounds

[0180]

[0181] Example 2: Mass spectrometry and chromatography detection

[0182] The chemical structure of some polypeptide compounds was detected using mass spectrometry and chromatography, and the specific detection results are shown in Table 3.

[0183] Table 3. Structure and detection data of polypeptide compounds

[0184]

[0185]

[0186]

[0187]

[0188] Example 3: Binding force detection of polypeptide compounds

[0189] The binding force of polypeptide compounds to the receptor protein GLP-1R was detected using surface plasmon resonance technology (SPR). 60 μL of GLP-1R protein with a concentration of 200 μg / mL was taken and added to 240 μL of sodium acetate buffer (10 mM, pH 4.5) and mixed well, and the final concentration of the protein was about 40 μg / mL. The CM5 chip was inserted into the device, and the protein was immobilized by amino coupling method. The dissociation rate constant (Kd) of the compound was measured by detecting the rate of separation of the target protein from each compound density. The detection results are shown in Table 4.

[0190] Table 4 SPR detection results of dissociation rate constant of polypeptide binding to GLP-1R

[0191] Sample Name Kd (pM) Semaglutide (positive control) 2.32 Dia2-001 17.0 Dia2-003 3.81 Dia2-004 2.98 Dia2-005 14.0 Dia2-006 4.53 Dia2-007 2.96 Dia2-008 2.58 Dia2-009 4.45 Dia2-010 1.21

[0192] As can be seen from the results of Table 3, the binding force of each polypeptide to GLP-1R is close, and the affinity is flat or better than Semaglutide, and the Kd value of Dia2-010 is the lowest, indicating that Dia2-010 has the strongest binding ability to GLP-1R.

[0193] Example 4: Detection of agonistic activity of polypeptide compounds on GLP-1R target

[0194] GLP-1 receptor (GLP-1R), as a member of the glucagon receptor subfamily of G protein-coupled receptor (GPCR) B family, GLP-1R agonists exert hypoglycemic effect in a blood glucose-dependent manner. When the blood glucose level in the body is elevated, GLP-1R agonists stimulate the release of insulin from pancreatic beta cells, lower blood glucose, and thus keep the blood glucose in the body at a relatively stable level, achieving the effect of treating diabetes. At the same time, GLP-1 agonists can inhibit gastrointestinal peristalsis and gastric juice secretion, reduce appetite, and delay gastric emptying, so as to produce a feeling of satiety in people, and ultimately achieve the effect of weight loss.

[0195] The agonistic activity of polypeptide compounds on GLP-1R target was detected using luciferase reporter gene method, which can be referred to the literature such as Jonathan W Day et al., Nat Chem Biol. 2009 Oct; 5(10): 749-57.

[0196] H_GLP1R-Reporter-HEK-293 cell line (Jiamin Biotech, Catalog No. GM-C25537) simultaneously expresses human GLP-1R gene and pCRE-Luc gene, which is a luciferase reporter gene cell line based on cAMP-PKA signaling pathway.

[0197] The specific experimental process is as follows:

[0198] The cells were cultured in a cell culture bottle with DMEM medium containing 10% FBS and 4 μg / mL Blasticidin and 0.75 μg / mL Puromycin, and when the confluence reached about 90%, the culture supernatant was discarded, 2 mL of trypsin was added for digestion, and then the digestion was terminated using DMEM medium containing 10% FBS, centrifuged at 1000 rpm for 5 min, the supernatant was discarded, 2 mL of DMEM medium containing 10% FBS was added for resuspension, and then the cells were counted. The cells were diluted with DMEM medium containing 10% FBS, 4 μg / mL Blasticidin and 0.75 μg / mL Puromycin, 100 μL was plated in each well of a 96-well plate, so that the cell density reached 1.5 x 10 4 / well, and then the polypeptide compounds, natural GLP-1 (MCE, 1013659) or semaglutide (Jiangxiao Biotechnology, CAS 910463-68-2) as a control were diluted to a series of specified concentrations with DMEM medium containing 1% FBS, added to the cell culture wells, 100 μL / well, and then detected after 4 h of stimulation. The detection was performed according to the instructions of the luciferase reporter assay kit (One GloTM Luciferase Assay Kit, Promega, Cat: E6120). The detection results are shown in Table 5, wherein the agonistic ability fitting curves of semaglutide and Dia2-001-010 are shown in Figures 1-8. Figure 17 .

[0199] Table 5 Experimental results of the agonistic ability of polypeptide compounds on H_GLP1R-Reporter-HEK-293 cells

[0200] Sample Name EC 50 (pmol / L)]]> Emax (%) Semaglutide (positive control) 146.5 107.9 Dia2-001 185.6 94.88 Dia2-002 30.8 110.8 Dia2-003 39.38 115.2 Dia2-004 164.4 102.9 Dia2-005 36.94 105.2 Dia2-006 61.99 105.9 Dia2-007 40.76 100.6 Dia2-008 208.5 97.24 Dia2-009 78.56 98.98 Dia2-010 169.9 98.44

[0201] As can be seen from Table 5 and Figure 17 , the EC 50 values of compounds Dia2-002, 003, 005-007 and 009 are less than that of semaglutide, and the maximum agonistic ability (Emax) is stronger for Dia2-003. This indicates that the polypeptides Dia2-002, 003, 005-007 and 009 are slightly better than semaglutide in the agonistic ability on GLP-1R.

[0202] Example 5: Effect of polypeptide compounds on blood glucose in mice

[0203] Male C57 / BL6 mice aged 8-10 weeks (purchased from Baishitong) were randomly selected, and the mice were bred under standard conditions and could freely obtain water and food, and were used for experiments after 5 days of adaptation.

[0204] Before the experiment, the mice were fasted overnight. On the next day, the mice were weighed before subcutaneous administration, and blood glucose was measured (Roche Accu-Chek Active, China) from the tail end, and the mice were randomly divided into groups (n = 5 or 6 per group) with similar initial average body weight and blood glucose. Then the mice were subcutaneously administered at 3-100 nM / kg, and the liquid dose was 1 mL / kg. After 3 hours of administration, blood glucose was measured from the tail end, and glucose was intraperitoneally injected (2 g / kg), and then the blood glucose values at 30, 60, and 120 minutes after glucose injection were measured, respectively, to calculate the blood glucose AUC (area under the curve, mmol*min / L) and evaluate the effect of the compounds on blood glucose.

[0205] After 240 minutes of administration, i.e., 60 minutes after glucose injection, the blood glucose values of each group, i.e., the AUC index, are shown in Table 6 below, and the changes in blood glucose after glucose injection in some groups are shown in FIG. 2. Figure 18

[0206] Table 6 Test results

[0207]

[0208]

[0209] As can be seen from the results in Table 6, the polypeptide compounds Dia2-001 to Dia2-010 can effectively reduce blood glucose, and among them, Dia2-004, 005, 006, and 010 are superior to the semaglutide group in terms of blood glucose data and AUC at 1 hour after glucose injection, indicating that they have better blood glucose regulation ability than semaglutide.

[0210] Example 6: Effect of polypeptide compounds on blood glucose and body weight of db / db mice

[0211] Male db / db mice aged 8-10 weeks were randomly selected and purchased from Guangdong Yaoke Biotechnology Co., Ltd. The mice were raised under standard conditions and had free access to water and food, and were used for experiments after 1 week of adaptation.

[0212] ​After the adaptation period, blood was randomly collected from the orbital cavity of the mice to measure blood glucose (Roche Accu-Chek Active, China). The cut-off values of blood glucose and body weight were 16.7 mmol / L and 42 g, respectively. That is, all mice participating in the experiment had blood glucose values higher than 16.7 mmol / L and body weights higher than 42 g, and the differences between the mice were small. At the beginning of the experiment, the mice were weighed, the blood glucose was measured, and they were randomly assigned to each treatment group (n = 5 / group) with similar average body weight and blood glucose at the start. Then the mice were subcutaneously administered 3-100 nM / kg, and the liquid dose was 1 mL / kg. The blood glucose was measured at 1, 3, 6, 24, and 48 hours, and the body weight was measured at 24 and 48 hours. The difference between the blood glucose and body weight values at each time point after administration and the blood glucose and body weight values before administration was calculated to evaluate the effect of the compound on body weight and blood glucose. The changes in blood glucose and body weight of the mice at 24 and 48 hours after administration are shown in Tables 7-8, Figure 19 and 20 .

[0213] Table 7 Changes in blood glucose of mice at 24 and 48 hours after administration

[0214] Dosing Group Change in blood glucose at 24 hours (mmol / L) Change in blood glucose at 48 hours (mmol / L) PBS (vehicle) 0.72±0.67 1.24±1.26 Semaglutide -13.13±1.80 -4.12±0.98 Dia2-001 -10.25±3.25 -0.30±1.93 Dia2-002 -11.95±1.47 4.68±1.82 Dia2-003 -17.94±1.55 -6.42±2.35 Dia2-004 -12.68±2.76 -6.55±4.41 Dia2-005 -14.08±1.62 2.55±3.52 Dia2-006 -10.63±2.76 -2.50±2.12

[0215] Table 8 Changes in body weight of db / db mice at 24 and 48 hours after administration

[0216]

[0217]

[0218] As can be seen from Tables 7 and 8, the polypeptide compounds Dia2-001-Dia2-006 all have the effect of reducing blood glucose, among which Dia2-002-Dia2-004 have a greater effect on reducing blood glucose at 48 h than semaglutide. The polypeptide compounds Dia2-002-Dia2-006 all have the effect of reducing body weight, among which Dia2-004 has a greater effect on weight loss at 48 h than semaglutide. In summary, the effects of compound Dia2-004 on reducing blood glucose and weight loss are greater than those of semaglutide 48 hours after administration, which indicates that Dia2-004 may have very good effects on long-term diabetic patients and obese people.

[0219] The above is only an embodiment of the present application, and is not a limitation in any form and in essence. It should be noted that, for those skilled in the art, without departing from the method of the present application, several improvements and supplements can also be made, which should also be considered as the protection scope of the present application. For those skilled in the art, without departing from the spirit and scope of the present application, some changes, modifications and equivalent changes made by using the above disclosed technical content are equivalent embodiments of the present application; at the same time, any equivalent changes, modifications and evolution of the above embodiments according to the essential technology of the present application are still within the scope of the technical solutions of the present application.

Claims

1. A GLP-1 polypeptide compound or a pharmaceutically acceptable salt, amide, or ester thereof, having an amino acid sequence of formula (I) as shown in SEQ ID NO:13: HX2EGTFTSDVSSYLEGQAAX 20 NFVX 24 X 25 LIARAEX 32 (I); in, X2 is aminoisobutyric acid; X 20 The K is a modified side chain, which is COOH-(CH2). a -CO-(γGlu) b -(NH-((CH2)2-O)2-CO)2-K, where a is any integer selected from 16-20, and b is 1; X 24 It is an amino acid selected from A, Q, or R; X 25 It is an amino acid selected from Q or R; X 32 It is AR 1 Or -R 1 R 1 It is a modification of the C-terminal carboxyl group of the sequence, wherein the modification is NH2 or is absent.

2. The GLP-1 polypeptide compound according to claim 1, or a pharmaceutically acceptable salt, amide, or ester thereof, wherein, a is 16 or 18.

3. The GLP-1 polypeptide compound according to claim 1 or 2, or a pharmaceutically acceptable salt, amide, or ester thereof, wherein the compound is selected from any one of the amino acid sequences shown in SEQ ID NO:1-12: (1) SEQ ID NO.1: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAE; (2) SEQ ID NO.2: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAE; (3) SEQ ID NO.3: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE; (4) SEQ ID NO.4: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE; (5) SEQ ID NO.5: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAE; (6) SEQ ID NO.6: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAE; (7)SEQ ID NO.7: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAEA; (8) SEQ ID NO.8: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAEA; (9)SEQ ID NO.9: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA; (10)SEQ ID NO.10: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA; (11)SEQ ID NO.11: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAEA; (12)SEQ ID NO.12: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAEA。 4. The GLP-1 polypeptide compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, amide, or ester thereof, wherein the compound is selected from any one of the following amino acid sequences: (1) SEQ ID NO.2: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVQRLIARAE; (2) SEQ ID NO.3: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE; (3) SEQ ID NO.4: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAE; (4) SEQ ID NO.5: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 16 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAE; (5) SEQ ID NO.6: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVARLIARAE; (6) SEQ ID NO.10: HAibEGTFTSDVSSYLEGQAA{COOH-(CH2) 18 -CO-γGlu-(NH-((CH2)2-O)2-CO)2-K}NFVRQLIARAEA。 5. The GLP-1 polypeptide compound according to any one of claims 1-4, or a pharmaceutically acceptable salt, amide, or ester thereof, wherein, The salt is a salt formed by a GLP-1 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-naphthylmethyl Acids, 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.

6. The GLP-1 polypeptide compound according to any one of claims 1-5, or a pharmaceutically acceptable salt, amide, or ester thereof, wherein, The salt is a salt formed by the GLP-1 polypeptide compound and any one of the following compounds: trifluoroacetic acid or acetic acid.

7. The GLP-1 polypeptide compound according to any one of claims 1-6, or a pharmaceutically acceptable salt, amide, or ester thereof, wherein its N-terminus or C-terminus is modified with an auxiliary group.

8. A pharmaceutical composition comprising the GLP-1 polypeptide compound of any one of claims 1-7 or a pharmaceutically acceptable salt, amide or ester thereof, and a pharmaceutically acceptable carrier, excipient or excipient; Preferably, the dosage form of the pharmaceutical composition is selected from any one of tablets, capsules, elixirs, syrups, lozenges, inhalers, sprays, injections, films, patches, powders, granules, blocks, emulsions, suppositories, or compound preparations.

9. Use of the GLP-1 polypeptide compound of any one of claims 1-7 or a pharmaceutically acceptable salt, amide or ester thereof, or the pharmaceutical composition of claim 8, in the preparation of a medicament for the prevention or treatment of metabolic syndromes such as diabetes, obesity, hypertension, dyslipidemia, atherosclerosis, coronary heart disease and other cardiovascular diseases, stroke, Alzheimer's disease, and non-alcoholic fatty liver disease.

10. A method for preparing the GLP-1 polypeptide compound of any one of claims 1-7 or a pharmaceutically acceptable salt, amide or ester thereof, wherein the preparation method is a chemical synthesis method.

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