Polypeptide compounds containing cyclic peptides, uses thereof and long-acting amylin medicaments

Abstract

The present application relates to the technical field of therapeutic cyclic peptide-containing polypeptides, in particular, to a cyclic peptide-containing polypeptide compound, application thereof and a long-acting amylin drug. The cyclic peptide-containing polypeptide compound is selected from the compounds shown in the following structural formula: wherein (Lactm bridge: Asp3-Lys8) represents an intramolecular lactam bridge formed between the side chain of the third amino acid Asp and the eighth amino acid Lys residue counted from the R1 end; R1 represents, R2 is independently selected from any one of carboxyl, carboxylate, tetrazole, phosphate, phosphate ester, sulfonic acid and sulfonate; n is any integer in 10-24; X 15 , X 16 , X 17 , X 22 , X 23 , X 24 , X 25 , X 26 and X 28 are independently selected from amino acids. As a novel amylin drug, the cyclic peptide-containing polypeptide compound is a multi-target drug, has good biological activity and bioavailability.

Description

Cross-references to related applications

[0001] This application claims priority to Chinese Patent Application No. CN202510911509.X, filed on July 2, 2025, entitled "Cyclic Peptide Compounds and Their Applications and Long-Acting Amylin Drugs", the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of therapeutic cyclic peptide technology, and more specifically, to cyclic peptide compounds, their applications, and long-acting amylin drugs. Background Technology

[0003] Human amylin is a 37-amino acid peptide hormone secreted and released by pancreatic β-cells, with the amino acid sequence: KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY. Amylin inhibits gastric emptying, controls appetite, and suppresses glucagon secretion, playing a crucial role in maintaining blood glucose levels. Furthermore, amylin promotes a feeling of fullness, preventing overeating and reducing energy intake; it also increases energy metabolism and reduces adipose tissue weight, making it a potential candidate drug for treating metabolic diseases such as obesity and diabetes. These metabolic regulatory functions of amylin are mediated by amylin receptors (AMYRS). Amylin receptors are dimer complexes formed by calcitonin receptors (CTRs) and receptor-modifying proteins (RAMPs).

[0004] However, the physicochemical properties of human amylin make it difficult to use as a drug. First, natural amylin is rapidly degraded by enzymes in the body and is quickly filtered and cleared by the kidneys, with an extremely short half-life of less than one hour. Therefore, patients need to administer the medication frequently during treatment, resulting in low adherence. Second, it is chemically unstable at physiological pH and easily precipitates, thus requiring preparation in acidic solutions. More importantly, amylin has a high tendency to fibrosis under physiological conditions, easily misfolding and agglomerating, leading to reduced drug target activity and stability, and even cytotoxicity.

[0005] Pramlintide is the only approved amylin analogue used as a complementary treatment for type 1 and type 2 diabetes. Due to the disulfide bridge (-SS-) formed by cysteine ​​residues at positions 2 and 7 and the deamidation effect of asparagine, prinlintide is unstable at neutral pH and is provided in acidic formulations. Furthermore, prinlintide has a half-life of only 30–50 minutes, requiring two to three injections daily, leading to patient inconvenience and poor adherence.

[0006] Cangrilintide is an investigational, novel, long-acting amylin analogue for the treatment of diabetes and obesity. Compared to human amylin, cangrilintide has amino acids at positions 25, 28, 29, and 37 replaced by proline, which reduces the drug's tendency to form fibrils. Simultaneously, γ-Glu-CO-(CH2) is introduced at position 1 (lysine). 18 The -CO2H side chain prolongs the drug's in vivo half-life to approximately 7 days. However, canagliflozin is unstable at neutral pH, with a stable formulation at pH 4.0, making it incompatible with neutral pH formulations of GLP-1 drugs. This complicates its co-use with these compounds.

[0007] The publicly disclosed amylintide analogue, Petrelintide (ZP8396), replaces the disulfide bridge (-SS-) formed by the cysteine ​​residues at positions 2 and 7 of prinlanpeptide and canagliflozin with a lactam bridge (-CO-NH-) formed by aspartic acid and lysine residues. Simultaneously, the aspartic acid at positions 21 and 22 of human amylintide, which is prone to deamidation, is removed, and N-methylated amino acids are introduced at positions 22 and 24, respectively. These structural modifications reduce the fibrotic tendency of Petrelintide, allowing it to be formulated at or near physiological pH. Petrelintide is administered subcutaneously and has a long in vivo half-life of approximately 10 days. Phase I clinical trials have shown good hypoglycemic and weight-loss efficacy; however, some patients taking therapeutic doses of Petrelintide experienced gastrointestinal side effects such as nausea, constipation, and diarrhea, and one subject discontinued treatment after the third treatment due to moderate nausea and vomiting.

[0008] Despite a global pipeline of 43 amyrin analogues, only prinlantide has been approved for marketing, and most of the compounds in development are in preclinical or Phase I clinical trials. There is significant clinical demand for AMYR-targeted drugs, indicating ample market potential. However, AMYR-targeted drugs such as Cagrilintide and Petrelintide still have serious gastrointestinal side effects, and all single-target amyrin analogues in development are administered subcutaneously. The pain at the injection site and "needle phobia" may reduce patient compliance, making it difficult to achieve ideal therapeutic effects.

[0009] In view of this, the present invention is proposed. Summary of the Invention

[0010] The purpose of this invention is to provide cyclic peptide-containing polypeptide compounds, their applications, and long-acting amylin drugs. The cyclic peptide-containing polypeptide compounds provided in the embodiments of this invention, as novel amylin drugs, are multi-target drugs with good biological activity and bioavailability.

[0011] This invention is implemented as follows: In a first aspect, the present invention provides a polypeptide compound containing a cyclic peptide, selected from compounds shown in the following structural formulas or their chiral isomers, tautomers, or pharmaceutically acceptable salts, esters, or amides:

[0012] Wherein, (Lactm bridge:Asp3-Lys8) means: an intramolecular lactam bridge formed between the side chain of the third amino acid Asp, counting from the R1 end, and the eighth amino acid Lys residue; R1 indicates R2 is independently selected from any one of carboxyl, carboxylic acid ester, tetrazolium, phosphoric acid, phosphate ester, sulfonic acid, and sulfonate groups; n is any integer from 10 to 24; X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 Each amino acid was selected independently.

[0013] Secondly, the present invention provides a long-acting amylin drug comprising a polypeptide compound containing a cyclic peptide as described in any of the foregoing embodiments.

[0014] Thirdly, the present invention provides the use of the cyclic peptide-containing polypeptide compound described in any of the foregoing embodiments in the preparation of a medicament for the prevention or treatment of any of the following diseases, said diseases including obesity, diabetes, insulin resistance syndrome, sleep apnea, obesity-induced cartilage degeneration and osteoarthritis, cardiovascular disease, macrovascular disease, microvascular disease, diabetic cardiomyopathy, coronary heart disease, myocardial infarction, stroke, arteriosclerosis, peripheral artery disease, gallbladder disease, non-alcoholic steatohepatitis, Alzheimer's disease, fatty liver disease, diabetic nephropathy, renal failure, hypertension, and heart failure.

[0015] The present invention has the following beneficial effects: The cyclic peptide-containing polypeptide compounds provided in the embodiments of the present invention can be used as amylin-like drugs, and are dual agonist drugs. They exhibit reduced fibrillation tendency and high chemical stability at physiological pH, possess good biological activity, and are suitable for various diseases. Simultaneously, they have a suitable long half-life and high bioavailability, which can reduce the frequency and dosage of administration, thereby improving patient compliance. Detailed Implementation

[0016] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0017] Amylin analogues have the potential to treat metabolic diseases such as obesity, diabetes, and hypertension, with as many as 43 amylin analogues currently in clinical trials. The only approved amylin analogue, prinlantide, has limited clinical efficacy and an extremely short half-life, far from meeting clinical needs. Furthermore, most amylin analogues under development are in preclinical or Phase I clinical trials and have significant gastrointestinal side effects. Therefore, developing amylin analogues with better bioactivity and safety would be a significant improvement to available treatment options. The cyclic peptide-containing polypeptide compound provided in this invention is also an AMYR-targeting compound. It not only possesses the aforementioned efficacy and can treat corresponding diseases, but it can also serve as a long-acting amylin drug, improving patient compliance.

[0018] Specifically, in a first aspect, embodiments of the present invention provide a polypeptide compound containing a cyclic peptide, selected from compounds shown in the following structural formulas or their chiral isomers, tautomers, or pharmaceutically acceptable salts, esters, or amides: Wherein, (Lactm bridge:Asp3-Lys8) means: an intramolecular lactam bridge formed between the side chain of the third amino acid Asp, counting from the R1 end, and the eighth amino acid Lys residue; R1 indicates R2 is independently selected from any one of carboxyl, carboxylic acid ester, tetrazolium, phosphate, phosphate ester, sulfonic acid, and sulfonate groups; n is any integer from 10 to 24; X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 Each amino acid was selected independently.

[0019] Furthermore, R1 represents R2 is independently selected from any one of carboxyl, carboxylic acid ester, tetrazolium, phosphate, phosphate ester, sulfonic acid, and sulfonate groups. n is any integer from 10 to 24; for example, n is any value between 10 and 24, such as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, and 24.

[0020] X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from Ser, N-Me-Ser and α-Me-Ser.

[0021] More preferably, R2 is independently selected from any one of carboxyl, tetrazolium, phosphate, and sulfonic acid groups; n is any integer from 14 to 24.

[0022] X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X28 It is selected from Ser, N-Me-Ser and α-Me-Ser.

[0023] Furthermore, R2 is independently selected from any one of carboxyl, tetrazolium, and phosphate groups; n is any integer from 16 to 20.

[0024] X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from Ser, N-Me-Ser and α-Me-Ser.

[0025] Furthermore, in the embodiments of the present invention, R2 is independently selected from any one of carboxyl, tetrazolium, and phosphate groups; n is 16, 18, or 20.

[0026] X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F), and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from Ser, N-Me-Ser, and α-Me-Ser. And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 They are not selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser, respectively.

[0027] Furthermore, in the embodiments of the present invention, R2 is independently selected from any one of carboxyl, tetrazolium, and phosphate groups; n is 16, 18, or 20.

[0028] X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from either Ser or N-Me-Ser. And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X28 They are not selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser, respectively.

[0029] Furthermore, in the embodiments of the present invention, R2 is independently selected from either a carboxyl group or a phosphate group; n is 16, 18, or 20.

[0030] X 15 Selected from any one of Aad, N-Me-Aad, Glu, and N-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from either Gly or N-Me-Gly; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from either Ser or N-Me-Ser. And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 X 28 They are not selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser, respectively.

[0031] Furthermore, in the embodiments of the present invention, R2 is independently selected from either a carboxyl group or a phosphate group; n is 18 or 20.

[0032] X 15 Selected from any one of Aad, N-Me-Aad, Glu, and N-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from either Gly or N-Me-Gly; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from either Ile or N-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from either Ser or N-Me-Ser. And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 They are not selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser, respectively.

[0033] Furthermore, in the embodiments of the present invention, R2 is independently selected from either a carboxyl group or a phosphate group; n is 18 or 20.

[0034] X 15 Choose either Aad or Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from either Gly or N-Me-Gly; X 24 Selected from any one of Ala, N-Me-Ala, and Aib; X 25 Selected from either Ile or N-Me-Ile; X 26 Selected from any one of Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from either Ser or N-Me-Ser. And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X28 They are not selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser, respectively.

[0035] Furthermore, in the embodiments of the present invention, the polypeptide compound is selected from any one of the compounds shown in the following structural formulas: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ; Preferably, the polypeptide compound is selected from any one of the compounds shown in the following structural formulas: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

[0036] Furthermore, embodiments of the present invention also provide a method for preparing the above-mentioned cyclic peptide-containing polypeptide compound. This preparation method can refer to existing methods. The embodiments of the present invention provide specific examples. This invention describes the synthesis of peptides via SPPS using a Fmoc-based chemical method on a peptide synthesis column. The Fmoc-protected amino acids used in this method were all purchased from Shanghai Jier Biochemical Co., Ltd., such as: Fmoc-Glu(OtBu)-OH, Fmoc-Glu-OtBu, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(OAll)-OH, Fmoc-Gly-OH, Fmoc- Thr(tBu)-OH, Fmoc-Ala-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Leu-OH, Fmoc-Aad(OtBu)-OH, Fmoc-Phe-OH, Fmoc-α-Me-Phe-OH, Fmoc-α-Me-Leu-OH, Fmoc-Gln(Trt)-OH, Fmoc-Ser(tBu)-OH, Fmoc-N-Me-Gly-OH, Fmoc-N-Me-Ile-OH, Fmoc-Val-OH, Fmoc- Asn(Trt)-OH, Fmoc-Hyp(tBu)-OH, Fmoc-N-Me- Phe -OH, Fmoc-N-Me- Leu -OH, Fmoc-α-Me- Glu(OtBu) -OH, Fmoc-N-Me- Glu(OtBu) -OH, Fmoc-Aib -OH, Fmoc-N-Me-Ser(tBu)-OH, Fmoc-α-Me-Ser(tBu)-OH.

[0037] Couplet: The resin used was Rink Amide-AM Resin crosslinked with 1% DVB from Jiangsu Haipu Functional Materials Co., Ltd., with a mesh size of 100-200 and a degree of substitution of 0.2-0.3 mmol / g. The Fmoc process was employed, with amino acids sequentially condensed and linked from the C-terminus to the N-terminus of the peptide chain. Rink Amide-AM Resin was added to a reaction vessel, DCM was added, and the mixture was stirred and swollen under nitrogen atmosphere. The protective amino acid solution (containing 0.1 M Oxyme) was used to remove the protective layer at 35°C. After the reaction was complete, the mixture was washed and filtered to obtain the resin to be reacted. Fmoc-Hyp(tBu)-OH and Oxyme were dissolved in NMP, and then DIC was added and mixed thoroughly. The activated protective amino acid solution was added to the resin to be reacted, and the mixture was stirred under nitrogen atmosphere at 60°C. After filtration and washing, the reaction was complete, yielding Fmoc. Hyp(tBu) AM Resin. Repeat the above steps, sequentially adding the protecting amino acids to the main chain. Before each subsequent coupling step, remove the Fmoc group using 20% ​​Pip / DMF (containing 0.1M Oxyme). All standard amino acid couplings were performed using a 2-fold molar ratio of Fmoc-protected amino acid, a 3-fold molar ratio of DIC, and a 3-fold molar ratio of HOBt, under nitrogen protection for 2–3 hours. The degree of coupling was detected using ninhydrin colorimetric reagent. In cases where coupling is difficult (e.g., methylated amino acids or other sterically hindered amino acids), increase the coupling time or repeat the coupling steps to achieve a satisfactory coupling level. After synthesis, wash the peptide resin with DCM and then air-dry thoroughly to obtain the peptide resin.

[0038] Lactam cyclization: After assembling the complete peptide sequence, in the DCM, the Asp (OAll) at position 3 and the Lys (Alloc) at position 8 were deprotected using tetrakis(triphenylphosphine)palladium(O) and phenylsilane. Subsequently, in the DMF, a lactam bridge was formed between the Asp residue at position 3 and the Lys residue at position 8 using HCTU and DIPEA.

[0039] Cutting: The synthesized dried peptide resin was cleaved at room temperature for 2 hours using a 10-fold volume lysis mixture (TFA:H2O:TIS, 95:2.5:2.5 v / v). The resin was filtered off, and the combined filtrates were treated with 5 fold volume of cold methyl ether (-20°C) to precipitate the crude peptide. The crude peptide / methyl ether suspension was then centrifuged at 3500 rpm for 10 minutes, the supernatant was removed, and the solid was washed twice with methyl ether and dried under vacuum to obtain the crude peptide.

[0040] purification: The crude peptide was dissolved in MQ water containing 40% acetic acid and purified by reversed-phase preparative HPLC (Waters Delta Prep 4000) on a C18 silica column. Elution was performed with MeCN in MQ water containing 0.1% TFA using a gradually increasing gradient. Fractions were analyzed by UPLC. Fractions containing the purified target peptide were combined. The resulting solutions were analyzed (UPLC, LCMS), and the product was aliquoted into glass vials. The vials were capped with a Millipore glass fiber pre-filter. Lyophilization yielded the target compound as a white solid.

[0041] General LCMS method: Mass spectrometry (MS) parameters: Detection instrument: Agilent 6410 triple quadrupole liquid chromatography-mass spectrometry system; Ionization source: +ESI.

[0042] Liquid chromatography parameters: Column: Agilent ZORBAX SB-C18, size: 250×4.6mm, 5μm; Detection wavelength: 195nm; Column temperature: 30℃; Injection volume: 10mL; Flow rate: 1mL / min; Mobile phase A: 0.05% glacial acetic acid; Mobile phase B: acetonitrile.

[0043] Secondly, the present invention also provides a long-acting amylin drug, which includes the polypeptide compound containing cyclic peptides described in the foregoing embodiments.

[0044] Thirdly, embodiments of the present invention also provide the use of the above-mentioned cyclic peptide-containing polypeptide compounds in medicaments for the prevention or treatment of any of the following diseases, including: obesity, diabetes, insulin resistance syndrome, sleep apnea, obesity-induced cartilage degeneration and osteoarthritis, cardiovascular disease, macrovascular disease, microvascular disease, diabetic cardiomyopathy, coronary heart disease, myocardial infarction, stroke, arteriosclerosis, peripheral artery disease, gallbladder disease, non-alcoholic steatohepatitis, Alzheimer's disease, fatty liver disease, diabetic nephropathy, renal failure, hypertension, and heart failure.

[0045] Furthermore, the names of the compounds or raw materials used in the preparation methods provided in the embodiments of the present invention, and their corresponding abbreviations, are as follows: Boc: tert-Butoxycarbonyl; Fmoc: 9-fluorenylmethoxycarbonyl; tBu: tert-Butyl; EDCl: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; HOSu: N-hydroxysuccinimide; TFA: trifluoroacetic acid; NMP: N-methylpyrrolidone; Pbf: 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl; DIC: diisopropylcarbodiimide; HCTU: 6-chlorophenyltriazole-1,1,3,3-tetramethylurea hexafluorophosphate; OAll: allyl ester; Alloc: allyloxycarbonyl; TIS: triisopropylsilane; Pd (PPh3)4: Tetra(triphenylphosphine)palladium(0); PhSiH3: Phenylsilane; Oxyme: Ethyl 2-oxime cyanoacetate; DCM: Dichloromethane; DMF: N,N-dimethylformamide; PIP: Piperidine; HOBt: 1-hydroxybenzotriazole; HBTU: Benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate; DIPEA: N,N-diisopropylethylamine; TIPS: Triisopropylsilane; Trt: Triphenylmethyl.

[0046] Furthermore, the non-natural amino acids used in the embodiments of the present invention and their corresponding structures are as follows: .

[0047] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0048] Example 1 This invention provides a cyclic polypeptide compound (hereinafter referred to as compound 1 PKWX202505-002), whose structural formula is as follows:

[0049] This invention also provides a method for preparing the above-mentioned polypeptide compound, which is synthesized according to the following synthetic route:

[0050] Specifically, A-1 (0.5 mmol) resin was added to a DCM (15 ml) solution, and the mixture was swelled and stirred under nitrogen for 2 hours, then the solvent was removed. 15 mL of 20% Pip / DMF (containing 0.1 M Oxyme) solution was added to remove the Fmoc protecting group, yielding 1-A2. Fmoc-Hyp(OtBu)-OH (1 mmol) and Oxyme (1.5 mmol) were dissolved in NMP, then DIC (1.5 mmol) was added for activation. 1-A2 was then added, and the mixture was stirred under nitrogen at 60 °C. After filtration and washing, 1-A3 was obtained. 1-A3 underwent amide condensation reactions sequentially with the fully protected amino acid and side chain fragment in the sequence to obtain 1-A4. In DCM solution, 1-B1 (1.35 mmol), HOSu (1.49 mmol), and EDCl·HCl (1.62 mmol) were added. The reaction was observed by TLC and, after completion, DIPEA (1.76 mmol) and compound 1-A4 (1.42 mmol) were added. After the reaction was complete, the mixture was washed and dried to obtain 1-A5. In DCM solution, 1-A5 was deprotected by tetrakis(triphenylphosphine)palladium(0) (0.25 mmol) and phenylsilane (1.0 mmol) to remove the Asp group at position 3 and the Lys side chain protecting group at position 8. Subsequently, a lactam bridge was formed between the Asp residue at position 3 and the Lys residue at position 8 using HCTU (0.9 mmol) and DIPEA (2.0 mmol). Following this, hydrogenation was performed to remove the benzyl group, yielding 1-A6. 1-A6 was cleaved at room temperature for 2 hours using a 10-fold volume lysis mixture (trifluoroacetic acid:water:triisopropylsilane, 95:2.5:2.5 v / v). The resin was filtered off and precipitated with methyl ether (-20°C). The precipitate was then centrifuged at 3500 rpm for 10 minutes, the supernatant was removed, and the solid was washed twice with methyl ether and dried under vacuum to obtain crude peptide 1-A7. Crude peptide 1-A7 was purified by reverse-phase chromatography and lyophilized to obtain target compound 1.

[0051] Compound 1 was characterized by: LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0052] Measured mass: (M+3) / 3=1410.10, (M+4) / 4=1057.86, (M+5) / 5=846.42.

[0053] Example 2-37 Examples 2-37 each provide a polypeptide compound containing a cyclic peptide. The preparation methods for Examples 2-37 are respectively carried out according to the preparation method provided in Example 1. The structural formulas and characterization data of the obtained polypeptide compounds containing cyclic peptides are as follows: Example 2: Compound 2 (PKWX202505-003), its structure is as follows: LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0054] Measured mass: (M+3) / 3=1398.03, (M+4) / 4=1048.89, (M+5) / 5=839.20.

[0055] Example 3: Compound 3 (PKWX202505-004), its structure is as follows:

[0056] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0057] Measured mass: (M+3) / 3=1398.13, (M+4) / 4=1048.79, (M+5) / 5=839.28.

[0058] Example 4: Compound 4 (PKWX202505-005), its structure is as follows:

[0059] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0060] Measured mass: (M+3) / 3=1398.15, (M+4) / 4=1048.74, (M+5) / 5=839.27.

[0061] Example 5: Compound 5 (PKWX202505-006), its structure is as follows:

[0062] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0063] Measured mass: (M+3) / 3=1410.13, (M+4) / 4=1057.79, (M+5) / 5=846.38.

[0064] Example 6: Compound 6 (PKWX202505-007), its structure is as follows:

[0065] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0066] Measured mass: (M+3) / 3=1410.09, (M+4) / 4=1057.75, (M+5) / 5=846.46.

[0067] Example 7: Compound 7 (PKWX202505-008), its structure is as follows:

[0068] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0069] Measured mass: (M+3) / 3=1410.03, (M+4) / 4=1057.84, (M+5) / 5=846.44.

[0070] Example 8: Compound 8 (PKWX202505-009), its structure is as follows:

[0071] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0072] Measured mass: (M+3) / 3=1410.06, (M+4) / 4=1057.72, (M+5) / 5=846.48.

[0073] Example 9: Compound 9 (PKWX202505-010), its structure is as follows:

[0074] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0075] Measured mass: (M+3) / 3=1405.76, (M+4) / 4=1054.52, (M+5) / 5=843.78.

[0076] Example 10: Compound 10 (PKWX202505-011), its structure is as follows:

[0077] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0078] Measured mass: (M+3) / 3=1405.70, (M+4) / 4=1054.57, (M+5) / 5=843.87.

[0079] Example 11: Compound 11 (PKWX202505-012), its structure is as follows:

[0080] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0081] Measured mass: (M+3) / 3=1398.05, (M+4) / 4=1048.80, (M+5) / 5=839.29.

[0082] Example 12: Compound 12 (PKWX202505-013), its structure is as follows:

[0083] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0084] Measured mass: (M+3) / 3=1410.01, (M+4) / 4=1057.76, (M+5) / 5=846.40.

[0085] Example 13: Compound 13 (PKWX202505-014), its structure is as follows:

[0086] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0087] Measured mass: (M+3) / 3=1398.05, (M+4) / 4=1048.79, (M+5) / 5=839.21.

[0088] Example 14: Compound 14 (PKWX202505-015), its structure is as follows:

[0089] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0090] Measured mass: (M+3) / 3=1398.00, (M+4) / 4=1048.84, (M+5) / 5=839.29.

[0091] Example 15: Compound 15 (PKWX202505-016), its structure is as follows:

[0092] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0093] Measured mass: (M+3) / 3=1410.03, (M+4) / 4=1057.72, (M+5) / 5=846.41.

[0094] Example 16: Compound 16 (PKWX202505-017), its structure is as follows:

[0095] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0096] Measured mass: (M+3) / 3=1405.68, (M+4) / 4=1054.58, (M+5) / 5=843.82.

[0097] Example 17: Compound 17 (PKWX202505-020), its structure is as follows:

[0098] LCMS Calculated mass: (M+3) / 3=1402.75, (M+4) / 4=1052.31, (M+5) / 5=842.05.

[0099] Measured mass: (M+3) / 3=1402.70, (M+4) / 4=1052.30, (M+5) / 5=842.12.

[0100] Example 18: Compound 18 (PKWX202505-021), its structure is as follows:

[0101] LCMS Calculated mass: (M+3) / 3=1414.41, (M+4) / 4=1061.06, (M+5) / 5=849.04.

[0102] Measured mass: (M+3) / 3=1414.43, (M+4) / 4=1061.00, (M+5) / 5=849.11.

[0103] Example 19: Compound 19 (PKWX202505-022), its structure is as follows:

[0104] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0105] Measured mass: (M+3) / 3=1398.05, (M+4) / 4=1048.83, (M+5) / 5=839.19.

[0106] Example 20: Compound 20 (PKWX202505-023), its structure is as follows:

[0107] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0108] Measured mass: (M+3) / 3=1410.00, (M+4) / 4=1057.84, (M+5) / 5=846.46.

[0109] Example 21: Compound 21 (PKWX202505-024), its structure is as follows:

[0110] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0111] Measured mass: (M+3) / 3=1410.09, (M+4) / 4=1057.86, (M+5) / 5=846.38.

[0112] Example 22: Compound 22 (PKWX202505-025), its structure is as follows:

[0113] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0114] Measured mass: (M+3) / 3=1398.14, (M+4) / 4=1048.89, (M+5) / 5=839.20.

[0115] Example 23: Compound 23 (PKWX202505-026), its structure is as follows:

[0116] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0117] Measured mass: (M+3) / 3=1410.10, (M+4) / 4=1057.75, (M+5) / 5=846.42.

[0118] Example 24: Compound 24 (PKWX202505-027), its structure is as follows:

[0119] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0120] Measured mass: (M+3) / 3=1410.04, (M+4) / 4=1057.73, (M+5) / 5=846.36.

[0121] Example 25: Compound 25 (PKWX202505-028), its structure is as follows:

[0122] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0123] Measured mass: (M+3) / 3=1405.70, (M+4) / 4=1054.48, (M+5) / 5=843.89.

[0124] Example 26: Compound 26 (PKWX202505-029), its structure is as follows:

[0125] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0126] Measured mass: (M+3) / 3=1405.82, (M+4) / 4=1054.59, (M+5) / 5=843.90.

[0127] Example 27: Compound 27 (PKWX202505-031), its structure is as follows:

[0128] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0129] Measured mass: (M+3) / 3=1398.04, (M+4) / 4=1048.86, (M+5) / 5=839.32.

[0130] Example 28: Compound 28 (PKWX202505-032), its structure is as follows:

[0131] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0132] Measured mass: (M+3) / 3=1410.13, (M+4) / 4=1057.76, (M+5) / 5=846.48.

[0133] Example 29: Compound 29 (PKWX202505-033), its structure is as follows:

[0134] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0135] Measured mass: (M+3) / 3=1398.15, (M+4) / 4=1048.75, (M+5) / 5=839.23.

[0136] Example 30: Compound 30 (PKWX202505-034), its structure is as follows:

[0137] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0138] Measured mass: (M+3) / 3=1410.05, (M+4) / 4=1057.73, (M+5) / 5=846.46.

[0139] Example 31: Compound 31 (PKWX202505-035), its structure is as follows:

[0140] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0141] Measured mass: (M+3) / 3=1405.82, (M+4) / 4=1054.50, (M+5) / 5=843.77.

[0142] Example 32: Compound 32 (PKWX202505-036), its structure is as follows:

[0143] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0144] Measured mass: (M+3) / 3=1405.76, (M+4) / 4=1054.48, (M+5) / 5=843.79.

[0145] Example 33: Compound 33 (PKWX202505-037), its structure is as follows:

[0146] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0147] Measured mass: (M+3) / 3=1398.10, (M+4) / 4=1048.89, (M+5) / 5=839.18.

[0148] Example 34: Compound 34 (PKWX202505-038), its structure is as follows:

[0149] LCMS Calculated mass: (M+3) / 3=1410.07, (M+4) / 4=1057.80, (M+5) / 5=846.44.

[0150] Measured mass: (M+3) / 3=1410.15, (M+4) / 4=1057.83, (M+5) / 5=846.50.

[0151] Example 35: Compound 35 (PKWX202505-039), its structure is as follows:

[0152] LCMS Calculated mass: (M+3) / 3=1405.75, (M+4) / 4=1054.56, (M+5) / 5=843.85.

[0153] Measured mass: (M+3) / 3=1405.87, (M+4) / 4=1054.65, (M+5) / 5=843.82.

[0154] Example 36: Compound 36 (PKWX202505-040), its structure is as follows:

[0155] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0156] Measured mass: (M+3) / 3=1398.18, (M+4) / 4=1048.72, (M+5) / 5=839.14.

[0157] Example 37: Compound 37 (PKWX202505-041), its structure is as follows:

[0158] LCMS Calculated mass: (M+3) / 3=1398.08, (M+4) / 4=1048.81, (M+5) / 5=839.25.

[0159] Measured mass: (M+3) / 3=1398.01, (M+4) / 4=1048.75, (M+5) / 5=839.36.

[0160] Test Example 1: hCTR / hAMYR3 agonist activity assay 1. Experimental Methods: In this experiment, HEK-293 cells stably expressing human calcitonin receptor (hCTR) or human amylin 3 receptor (hAMYR3) were used. The cAMP content was detected by TR-FRET method to evaluate the hCTR / hAMYR3 receptor activity of the compounds of this invention.

[0161] 2. Experimental procedure: 1) HEK-293 cells stably expressing hCTR or hAMYR3 were digested with trypsin, centrifuged, resuspended and counted, and seeded at 10,000 cells per well in 384-well cell plates and incubated overnight at 37°C and 5% CO2.

[0162] 2) Prepare appropriate gradient working concentrations of Petrelintide (prepared according to the method described in patent CN109863168B) and the compound of the present invention using assay buffer, with a total of 10 dilutions, each in duplicate.

[0163] 3) Following the experimental plate layout, add serially diluted samples to a 384-well cell culture plate and incubate at 37°C and 5% CO2 for 1 h. Measure cAMP levels using a cAMP kit (PerkinElmer) according to the manufacturer's recommendations and the protocol. Calculate EC50 using GraphPadPrism curve fitting. 50 value.

[0164] 3. The experimental results are shown in Table 1: Table 1 Results of agonist activity tests on the compounds

[0165] 4. Conclusion: The compounds provided in the embodiments of this invention exhibit strong agonistic effects on both hCTR and hAMYR3. Specifically, the compounds provided in the embodiments of this invention have superior agonistic activity against hCTR and hAMYR3 compared to Petrelintide, and some of the compounds in the embodiments show more than 10 times higher agonistic activity against hCTR and hAMYR3 than Petrelintide, thus demonstrating better potential for blood sugar reduction and weight loss.

[0166] Test Example 2: Detection Test for the Tendency to Form Insoluble Amyloid Fibers 1. Experimental method: This experiment uses thioflavin T (ThT) and the physical stability of the compound of the present invention is detected by fluorescence assay.

[0167] 2. Experimental procedure: 1) Dissolve Cagrilintide (purchased from Jiangsu Aikon Biomedical R&D Co., Ltd.), Petrelintide, and the compound of this invention in a buffer solution (sodium acetate at pH 4.0 or sodium phosphate at pH 7.0) to a concentration of 250 μM and incubate for 1 h. Then add ThT stock solution to obtain a sample with a final ThT concentration of 1 μM.

[0168] 2) According to the experimental plate layout, add 200 μL of sample to each well of a transparent bottom black fluorescent 96-well plate, with 5 replicates for each sample, and seal with a sealing film.

[0169] 3) Incubate under specific conditions (40℃, 960 rpm, 1 mm amplitude) with shaking. Measure fluorescence intensity every 20 min using 444 nm excitation and 480 nm emission light for 96 h. The physical stability of the test compound can be assessed by detecting changes in fluorescence signal using a microplate reader.

[0170] 3. The experimental results are shown in Table 2: Table 2. Tendency of compounds to form amyloid fibrils in buffer solution

[0171] Note: "-" indicates that insoluble amyloid fibers were not detected; "+" indicates that insoluble amyloid fibers were detected. 4. Conclusion: Cagrilintide formed insoluble amyloid fibers within 48 h in both sodium acetate (pH 4) and sodium phosphate (pH 7) buffer solutions. The compounds provided in this embodiment of the invention did not form insoluble amyloid fibers in either buffer solution for 48 h, and only a very small amount formed after 96 h, demonstrating generally good physical stability.

[0172] Test Example 3: Chemical Stability Test 1. Experimental steps: 1) Dissolve the compound to be tested in sodium acetate (50 nM) buffer at pH 4 or sodium phosphate (50 nM) buffer at pH 7 to a final concentration of 1 mg / mL, and incubate at 40 °C.

[0173] 2) The sample was eluted on a C18 column using a gradient elution buffer of trifluoroacetic acid / acetonitrile / water, and the sample was analyzed by reversed-phase high-performance liquid chromatography.

[0174] 3) The percentage of the main peak area at each sampling time point minus the initial percentage of the main peak area is the degradation.

[0175] 2. The experimental results are shown in Table 3: Table 3. Degradation % of compounds in buffer solution

[0176] 3. Conclusion: The compounds provided in this invention exhibit good chemical stability, with a degradation rate comparable to Petrelintide after 72 h in sodium acetate buffer at pH 4 and sodium phosphate buffer at pH 7.

[0177] Test Example 4: Acute Effects of Food Intake and Body Weight on Normal Rats 1. Experimental objective: The effects of the compounds provided in the embodiments of this invention on acute food intake and body weight in normal Sprague Dawley (SD) rats were investigated and compared with Petrelintide.

[0178] 2. Animal selection: Adult Sprague Dawley (SD) rats were selected as experimental animals, and the animals were acclimatized for at least one week before the start of the study.

[0179] 3. Grouping of experimental animals and drug intervention measures: Animals were randomly divided into groups of eight. Each animal was then administered a single subcutaneous injection, either the solvent, 30 nmol / kg of Petrelintide, or 30 nmol / kg of the compound of this invention.

[0180] 4. Measurement of food intake and body weight: Throughout the study, the animals had free access to food and water. Food intake and body weight were recorded at 12 h, 24 h, 36 h, and 48 h after feeding. Data processing was performed using GraphPadPrism.

[0181] 5. The experimental results are shown in Table 4: Table 4. Changes in food intake (%, compared to the solvent group) and body weight (%, compared to the solvent group) of SD rats over 48 h.

[0182] Note: "-" indicates a decrease; 6. Conclusion: During the experiment, the rats were in good health and mental condition. The compounds provided in the embodiments of this invention all exhibited good acute food intake inhibition and weight loss in normal rats. Specifically, the compounds provided in the embodiments of this invention showed better acute food intake inhibition and weight loss than Petrelintide in normal rats.

[0183] Test Example 5: Changes in body weight and blood glucose in ZDF rats after acute feeding 1. Experimental objective: The study investigated the changes in body weight and blood glucose in ZDF rats after acute feeding of the compounds provided in the embodiments of the present invention, and compared them with Petrelintide.

[0184] 2. Selection of animal models: The experimental animals selected were ZDF rats (Zucker Diabetic Fatty rats), a type 2 diabetes animal model characterized by obesity and severe hyperglycemia, and one of the most commonly used and classic models for studying type 2 diabetes and its complications. Animals underwent at least one week of acclimatization before the start of the study.

[0185] 3. Grouping of experimental animals and drug intervention measures: ZDF rats were randomly divided into groups of eight. According to the group, each animal was given a single subcutaneous injection of the solvent, 10 nmol / kg of Petrelintide, or 10 nmol / kg of the compound of this invention.

[0186] 4. Weight and blood glucose measurement: Throughout the study, the animals had free access to food and water. Body weight was recorded in ZDF rats before administration and at 12 h, 24 h, and 48 h after administration. Tail blood samples were collected before administration and at 12 h, 24 h, and 48 h after administration, and blood glucose levels (mmol / L) were measured using a glucometer.

[0187] 5. The experimental results are shown in Table 5: Table 5. Changes in body weight (%, compared to baseline) and changes in glucose (%, compared to baseline) in ZDF rats

[0188] Note: "-" indicates a decrease; 6. Conclusion: During the experiment, the rats were in good health and mental condition. A single subcutaneous injection of the compound described in this invention demonstrated a superior effect on reducing body weight and blood glucose levels in ZDF rats compared to Petrelintide.

[0189] Test Example 6: Rat Pharmacokinetic (PK) Test 1. Experimental objective: The pharmacokinetic parameters of the compounds provided in the embodiments of the present invention in Sprague Dawley (SD) rats were studied and compared with those of Petrelintide.

[0190] 2. Animal selection: Adult Sprague Dawley (SD) rats were selected as experimental animals, and the animals were acclimatized for at least one week before the start of the study.

[0191] 3. Grouping of experimental animals and drug intervention measures: Animals were randomly divided into groups of eight. Each animal was then given a single subcutaneous injection of either 30 nmol / kg Petrelintide or 30 nmol / kg of the compound of this invention, according to its group.

[0192] 4. Blood sample collection and analysis: Throughout the study, animals had free access to food and water. Venous blood samples were collected before administration and at 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 48 h, 72 h, and 96 h after administration. Blood samples were placed on ice, and plasma was separated by centrifugation within 1 h and analyzed using LC-MS / MS.

[0193] 5. The experimental results are shown in Table 6: Table 6 Pharmacokinetic parameters of SD rats

[0194] 6. Conclusion: The compounds provided in the embodiments of this invention all have a longer half-life in rats than Petrelintide, and can be used for the development of long-acting formulations.

[0195] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A polypeptide compound containing a cyclic peptide, characterized in that, It is selected from compounds shown in the following structural formulas or their chiral isomers, tautomers, or pharmaceutically acceptable salts, esters, or amides: Wherein, (Lactm bridge:Asp3-Lys8) means: an intramolecular lactam bridge formed between the side chain of the third amino acid Asp, counting from the R1 end, and the eighth amino acid Lys residue; R1 indicates R2 is independently selected from any one of carboxyl, carboxylic acid ester, tetrazolium, phosphoric acid, phosphate ester, sulfonic acid, and sulfonate groups; n is any integer from 10 to 24; X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 Each amino acid was selected independently.

2. The polypeptide compound containing a cyclic peptide according to claim 1, characterized in that, R1 indicates R2 is independently selected from any one of carboxyl, carboxylic acid ester, tetrazolium, phosphoric acid, phosphate ester, sulfonic acid, and sulfonate groups; n is any integer from 10 to 24; X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from Ser, N-Me-Ser and α-Me-Ser; Preferably, R2 is independently selected from any one of carboxyl, tetrazolium, phosphoric acid, and sulfonic acid groups; n is any integer from 14 to 24; X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from Ser, N-Me-Ser and α-Me-Ser; Preferably, R2 is independently selected from any one of carboxyl, tetrazolium, and phosphate groups; n is any integer from 16 to 20; X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 It is selected from Ser, N-Me-Ser and α-Me-Ser.

3. The polypeptide compound containing a cyclic peptide according to claim 1 or 2, characterized in that, R2 is independently selected from any one of carboxyl, tetrazolium, and phosphate groups; n is 16, 18, or 20; X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, α-Me-Phe, α-Me-Phe(2-F) and α-Me-Phe(4-F); X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from Ser, N-Me-Ser and α-Me-Ser; And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 They are not simultaneously selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser.

4. The polypeptide compound containing a cyclic peptide according to claim 1 or 2, characterized in that, R2 is independently selected from any one of carboxyl, tetrazolium, and phosphate groups; n is 16, 18, or 20; X 15 Selected from any one of Aad, N-Me-Aad, Glu, N-Me-Glu, and α-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from Gly, N-Me-Gly, and Ala; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from either Ser or N-Me-Ser; And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 They are not simultaneously selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser.

5. The polypeptide compound containing a cyclic peptide according to claim 1 or 2, characterized in that, R2 is independently selected from either a carboxyl group or a phosphate group; n is 16, 18, or 20; X 15 Selected from any one of Aad, N-Me-Aad, Glu, and N-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from either Gly or N-Me-Gly; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Selected from any one of Ile, N-Me-Ile, and α-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from either Ser or N-Me-Ser; And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 X 28 They are not simultaneously selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser.

6. The polypeptide compound containing a cyclic peptide according to claim 1 or 2, characterized in that, R2 is independently selected from either a carboxyl group or a phosphate group; n is 18 or 20; X 15 Selected from any one of Aad, N-Me-Aad, Glu, and N-Me-Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from either Gly or N-Me-Gly; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Choose either Ile or N-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from either Ser or N-Me-Ser; And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 They are not simultaneously selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser.

7. The polypeptide compound containing a cyclic peptide according to claim 1 or 2, characterized in that, R2 is independently selected from either a carboxyl group or a phosphate group; n is 18 or 20; X 15 Choose either Aad or Glu; X 16 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 17 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 22 Selected from any one of Phe, N-Me-Phe, and α-Me-Phe; X 23 Selected from either Gly or N-Me-Gly; X 24 Selected from Ala, N-Me-Ala, and Aib; X 25 Choose either Ile or N-Me-Ile; X 26 Selected from Leu, N-Me-Leu, and α-Me-Leu; X 28 Selected from either Ser or N-Me-Ser; And when R2 is a carboxyl group and n=18, X 15 X 16 X 17 X 22 X 23 X 24 X 25 X 26 and X 28 They are not simultaneously selected from Aad, Phe, Leu, Phe, N-Me-Gly, Ala, N-Me-Ile, Leu, and Ser.

8. The polypeptide compound containing a cyclic peptide according to claim 1 or 2, characterized in that, The polypeptide compound is selected from any one of the compounds shown in the following structural formulas: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 or ； Preferably, the polypeptide compound is selected from any one of the compounds shown in the following structural formulas: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 or 。 9. A long-acting amylin drug, characterized in that, It includes the polypeptide compound containing a cyclic peptide as described in any one of claims 1-8.

10. The use of a polypeptide compound containing a cyclic peptide according to any one of claims 1-8 in the preparation of a medicament for the prevention or treatment of any of the following diseases, characterized in that, The diseases mentioned include any one of the following: obesity, diabetes, insulin resistance syndrome, sleep apnea, obesity-induced cartilage degeneration and osteoarthritis, cardiovascular disease, large vessel disease, microvascular disease, diabetic cardiomyopathy, coronary heart disease, myocardial infarction, stroke, arteriosclerosis, peripheral artery disease, gallbladder disease, non-alcoholic steatohepatitis, Alzheimer's disease, fatty liver disease, diabetic nephropathy, renal failure, hypertension, and heart failure.

Images