Pharmaceutical use of polypeptides in the preparation of medicaments for treating and / or preventing diabetes, obesity and related disorders

Abstract

The pharmaceutical use of a series of polypeptides in the preparation of medicaments for treating and / or preventing diabetes, obesity and related diseases, in particular the polypeptide having the sequence of formula (II): YAibEGT FTSDY SIAibLD KIAQK AFVKW LIAGG PSSGA PPPS0(II)

Description

[Technical field]

[0001] The present invention belongs to the biomedical field and relates to the pharmaceutical use of a polypeptide whose sequence is shown in formula (II) in the preparation of a medicament for the treatment and / or prevention of diabetes, obesity and related diseases. [Background technology]

[0002] The global prevalence of nonalcoholic fatty liver disease (NAFLD) is as high as 25%, and the global prevalence of nonalcoholic steatohepatitis (NASH) is approximately 3% to 8%. Some patients with NASH further progress to liver sclerosis and liver cancer, which is currently one of the leading causes of end-stage liver disease and liver transplantation. The pathogenesis of NASH is complex, and there is currently no FDA-approved drug for the disease. Several preclinical studies have shown that glucose-dependent insulinotropic polypeptide (GIP) / glucagon-like peptide-1 (GLP-1) dual agonists can be used to treat NASH. Clinical studies of the GLP-1 / GIP dual agonist Tirzepatide, a drug under investigation by Eli Lilly and Company, have shown that it improves related markers such as NASH-related transaminases, indicating its potential as a treatment for NASH.

[0003] GLP-1 agonists can treat NASH through multi-pathway synergistic effects. In vivo studies have found that GLP-1 agonists reduce body weight by suppressing appetite in the brain, delaying gastric emptying, reducing hepatic gluconeogenesis, and increasing glucose consumption in muscle tissue. At the same time, GLP-1 agonists reduce circulating levels of tumor necrosis factor (TNF)-α, interleukin IL-1β, IL-6, CD163, and hsCRP, achieving anti-inflammatory effects. In addition, GLP-1 agonists can improve NASH symptoms through multi-pathway by inhibiting fatty acid synthase, suppressing LDL uptake, and reducing adipose tissue blood flow. GIP is a polypeptide secreted from neuroendocrine K cells in the small intestine, and activating GIPR can promote lipid metabolism, which can further alleviate hepatic lipid synthesis based on GLP-1 agonist treatment, so GLP-1 / GIP dual agonists have a synergistic effect in the treatment of NASH.

[0004] During the course of research, it was also discovered that GLP-1 / GIP dual agonists have excellent effects on regulating the body's glucose metabolism and lipid metabolism, and compounds targeting this target have been shown to be promising for development as preventive and / or therapeutic drugs for various specific diseases accompanied by glucose metabolism disorders and / or lipid metabolism disorders, such as diabetes, obesity, and complications (related diseases) resulting from these diseases. Summary of the Invention

[0005] The present invention relates to a method for producing a compound having a sequence of formula (II), i.e. YAibEGT FTSDY SIAibLD KIAQK AFVKW LIAGG PSSGA PPPS0 (II) is shown in As shown below: 1) The amino groups on the lysine side chain at positions i and i+3 or the amino groups on the lysine side chain at positions j and j+4 TIFF2025512980000001.tif10170, where i is 17 (i.e., when i is 17, first the isoleucine at position 17 is replaced with lysine, and then the amino group on its side chain is linked to the amino group on the lysine side chain at position 20, where the amino groups on the lysine side chains at positions 17 and 20 are each linked to X, and the amino group is TIFF2025512980000002.tif10170), or where j is 20, and 2) an additional 0 to 2 amino acids of the polypeptide of formula (II) are substituted; Where: The structure of Aib is: TIFF2025512980000003.tif15170, S0 is a structure TIFF2025512980000004.tif17170, serine (S,(S)-2-amino-3-hydroxypropionic acid) and TIFF2025512980000005.tif17170, (S)-2-amino-3-hydroxypropionamide (i.e., the amino acid at position 39 (serine) is optionally amidated to a C-terminal primary amide); X is Selected from TIFF2025512980000006.tif17170, where "*" represents the position concatenated to X1, X1 is selected from a single bond, -C(=O)-, -OC(=O)- and -N(R1)-C(=O)-; R1 is H and C 1-3 alkyl, X2 is Selected from TIFF2025512980000007.tif38170, m is selected from 2, 3 and 4; n is selected from 15, 16, 17, 18 and 19; p includes a modification selected from 1 and 2 The pharmaceutical use of the polypeptide in the preparation of a medicament for treating and / or preventing diabetes, obesity and related disorders is provided.

[0006] The polypeptide sequence whose sequence is shown in formula (II) is as shown in Seq ID NO.1.

[0007] The present invention relates to a method for producing a compound having a sequence of formula (P): YAibEGT FTSDY SIAibLD KKAQK AFVKW LIAGG PSSGA PPPS0 (P) is shown in As shown below: 1) The amino groups on the lysine side chains at positions 17 and 20 Concatenated to TIFF2025512980000008.tif10170, and 2) 0 to 2 amino acids of a polypeptide having a sequence represented by formula (P) are substituted, Where: The structure of Aib is TIFF2025512980000009.tif15170, S0 is a structure TIFF2025512980000010.tif18170, serine (S,(S)-2-amino-3-hydroxypropionic acid), and the structure TIFF2025512980000011.tif18170, (S)-2-amino-3-hydroxypropionamide (i.e., the amino acid at position 39 (serine) is optionally amidated to a C-terminal primary amide); X is Selected from TIFF2025512980000012.tif17170, where "*" represents the position concatenated to X1, X1 is selected from a single bond, -C(=O)-, -OC(=O)- and -N(R1)-C(=O)-; R1 is H and C 1-3 alkyl, X2 is Selected from TIFF2025512980000013.tif38170, m is selected from 2, 3 and 4; n is selected from 15, 16, 17, 18 and 19; p includes a modification selected from 1 and 2 The pharmaceutical use of the polypeptide in the preparation of a medicament for treating and / or preventing diabetes, obesity and related disorders is provided.

[0008] The polypeptide sequence whose sequence is represented by formula (P) is as set forth in Seq ID NO.2.

[0009] In some aspects of the invention, 0 to 2 amino acids at positions 21, 23 or 24 of the above polypeptides are substituted, with the remaining variables being as defined herein.

[0010] In some aspects of the present invention, the polypeptide has a sequence selected from the sequences represented by formulas (II-1), (II-2), (II-3), (II-4), (III-6), (IV-7), (IV-8), (IV-9) and (IV-10), and the polypeptides represented by formulas (II-1), (II-2), (II-3), (II-4), (III-6), (IV-7), (IV-8), (IV-9) and (IV-10) are represented by Seq ID NOs. 3 to 11, respectively. YAibEGT FTSDY SIAibLD KIAQK AFVKW LIAGG PSSGA PPPS-NH2(II-1) YAibEGT FTSDY SIAibLD KIAQK EFVKW LIAGG PSSGA PPPS-NH2(II-2) YAibEGT FTSDY SIAibLD KIAQK AFIKW LIAGG PSSGA PPPS-NH2(II-3) YAibEGT FTSDY SIAibLD KIAQK AFVKW LLAGG PSSGA PPPS-NH2(II-4) YAibEGT FTSDY SIAibLD KKAQK AFVEW LIAGG PSSGA PPPS-NH2(III-6) YAibEGT FTSDY SIAibLD KKAQK AFVQW LIAGG PSSGA PPPS-NH2(IV-7) YAibEGT FTSDY SIAibLD KKAQK AFVAW LIAGG PSSGA PPPS-NH2(IV-8) YAibEGT FTSDY SIAibLD KKAQK AFVIW LIAGG PSSGA PPPS-NH2(IV-9) YAibEGT FTSDY SIAibLD KKAQK EFVEW LIAGG PSSGA PPPS-NH2(IV-10) As shown below: The amino groups on the lysine side chains at positions 17 and 20 or the amino groups on the lysine side chains at positions 20 and 24 Concatenated into TIFF2025512980000014.tif10170, wherein the serine at position 39 is amidated to a C-terminal primary amide, and Aib, X, X1 and X2 are as defined herein.

[0011] In some aspects of the invention, R1 above is selected from H, with the remaining variables being as defined herein.

[0012] In some aspects of the invention, X1 above is selected from a single bond, -C(=O)-, -OC(=O)-, and -NH-C(=O)-, with the remainder of the variables being as defined herein.

[0013] In some aspects of the invention, m above is selected from 2, with the remaining variables being as defined herein.

[0014] In some aspects of the invention, n above is selected from 15 and 17, with the remaining variables being as defined herein.

[0015] In some aspects of the present invention, X2 is TIFF2025512980000015.tif76170, and the remaining variables are as defined herein.

[0016] In some aspects of the present invention, X2 is TIFF2025512980000016.tif75170, and the remaining variables are as defined herein.

[0017] In some aspects of the invention, the amino groups on the lysine side chains at positions i and i+3 or the amino groups on the lysine side chains at positions j and j+4 are Link to TIFF2025512980000017.tif10170, TIFF2025512980000018.tif93170, with the remaining variables as defined herein.

[0018] In some aspects of the invention, the amino groups on the lysine side chains at positions i and i+3 or the amino groups on the lysine side chains at positions j and j+4 are Link to TIFF2025512980000019.tif10170, TIFF2025512980000020.tif92170, with the remaining variables as defined herein.

[0019] In some aspects of the invention, TIFF2025512980000021.tif10170 is TIFF2025512980000022.tif176170, and the remaining variables are as defined herein.

[0020] In some aspects of the invention, TIFF2025512980000023.tif10170 is TIFF2025512980000024.tif122170, and the remaining variables are as defined herein. The present invention also has several aspects derived from any combination of the above variables.

[0021] The present invention further provides the pharmaceutical use of a polypeptide selected from those represented by the formula below in the preparation of a medicament for treating and / or preventing diabetes, obesity and related diseases. TIFF2025512980000025.tif234170TIFF2025512980000026.tif240170TIFF2025512980000027.tif245170

[0022] The present invention further provides the use of a series of pharmaceutical compositions comprising a therapeutically effective amount of the above-mentioned polypeptide compound or a pharma- ceutically acceptable salt thereof as an active ingredient, and a pharma- ceutically acceptable carrier, in the preparation of medicaments for preventing and / or treating diabetes, obesity and related comorbid diseases (associated diseases).

[0023] Diabetes is a chronic disease with many pathological symptoms, a metabolic and endocrine disease caused by absolute insulin deficiency or reduced biological effect of insulin, and is one of the specific indications of lipid metabolism disorder, circulatory disorder and / or glucose metabolism disorder in the body. Diabetes is prone to cause various complications (related diseases) such as diabetic cardiovascular disease, diabetic cerebrovascular disease, retinopathy, nephropathy, cataract, coronary artery disease, diabetic neuropathy, diabetic foot, and peripheral neuropathy. Among these, diabetic cardiovascular disease complications include microvascular disease on the heart and large blood vessels, cardiomyopathy, and cardiac autonomic neuropathy, and diabetic cerebrovascular disease includes cerebral arteriosclerosis, ischemic cerebrovascular disease, cerebral hemorrhage, and cerebral atrophy.

[0024] Obesity is another common metabolic syndrome caused by an accumulation of unhealthy diet, lack of exercise, emotional factors, genetic factors, etc., and is also a specific indication of lipid metabolism disorders, circulatory disorders, and / or glucose metabolism disorders in the body. Meanwhile, obesity increases the risk of developing related diseases such as liver disease, hyperlipidemia, diabetes, and hypertension.

[0025] Diabetes, obesity and related diseases have significant impacts on human health and socio-economic outcomes.

[0026] In some aspects of the invention, there is provided the use of the above-described polypeptide compound or a pharma- ceutically acceptable salt thereof, or the above-described pharmaceutical composition in the preparation of a medicament for treating diabetes and / or obesity.

[0027] In some aspects of the present invention, there is provided the above-mentioned polypeptide compound or a pharma- ceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition, which is administered once every 3 days, once every 4 days, once every week, or once every 2 weeks. Technical effects

[0028] The polypeptide of the present invention has very strong agonistic activity against GLP-1R / GIPR, and the polypeptide compound of the present invention has excellent pharmacokinetic properties, plasma stability and extremely high plasma protein binding ability. Definitions and Explanations

[0029] Unless otherwise specified, the following terms and phrases used herein shall have the following meanings: Certain terms or phrases, unless otherwise defined, should not be considered unclear or unclear, but should be understood in their general sense. When a trade name is mentioned in this specification, it is intended to refer to the corresponding product or its active ingredient.

[0030] As used herein, the term "pharmacologically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic response, other problem or complication, and commensurate with a reasonable benefit / risk ratio.

[0031] The term "pharmaceutical acceptable salt" refers to a salt of the polypeptide compound of the present invention prepared from a compound having a specific substituent found in the present invention and a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting such a compound with a sufficient amount of base in a pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium salts, potassium salts, calcium salts, ammonium salts, organic amine or magnesium salts or similar salts. When the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting such a compound with a sufficient amount of acid in a pure solution or in a suitable inert solvent. Examples of pharma- ceutically acceptable acid addition salts include inorganic acid salts, including hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like, and organic acid salts, including acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanedioic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like, as well as salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain compounds of the present invention contain both basic and acidic functional groups and therefore can be converted into either base or acid addition salts.

[0032] The pharma- ceutically acceptable salts of the present invention can be synthesized from parent compounds containing acids or bases using conventional chemical methods. In general, the method for preparing such salts is by reacting the compounds in their free acid or base form with a stoichiometric amount of an appropriate base or acid in water, an organic solvent, or a mixture thereof.

[0033] "Amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to naturally occurring amino acids. Naturally occurring amino acids are those amino acids encoded by the genetic code, as well as those amino acids that have been subsequently modified, such as hydroxyproline, gamma-carboxyglutamine, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure (e.g., an alpha carbon bound to a hydrogen, a carboxyl group, an amino group, and an R group) as a naturally occurring amino acid, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs may have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refer to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions similarly to a naturally occurring amino acid.

[0034] As used herein, A or Ala has the structure TIFF2025512980000028.tif15170 represents alanine, and R or Arg represents TIFF2025512980000029.tif18170 represents arginine, and N or Asn represents the structure TIFF2025512980000030.tif16170 represents asparagine, and D or Asp represents TIFF2025512980000031.tif16170 represents aspartic acid, and C or Cys represents TIFF2025512980000032.tif16170 represents cysteine, and Q or Gln represents TIFF2025512980000033.tif16170 represents glutamine, and E or Glu represents TIFF2025512980000034.tif16170 represents glutamic acid, and G or Gly has the structure TIFF2025512980000035.tif11170 represents glycine, and H or His represents TIFF2025512980000036.tif18170 represents histidine, and I or Ile represents the structure TIFF2025512980000037.tif19170 represents isoleucine, and L or Leu represents TIFF2025512980000038.tif16170 represents leucine, and K or Lys represents leucine, whose structure is TIFF2025512980000039.tif17170 represents lysine, and M or Met represents TIFF2025512980000040.tif17170 represents methionine, and F or Phe represents TIFF2025512980000041.tif17170 represents phenylalanine, and P or Pro represents TIFF2025512980000042.tif17170 represents proline, and S or Ser represents TIFF2025512980000043.tif17170 represents serine, and T or Thr represents TIFF2025512980000044.tif17170 represents threonine, and W or Trp represents TIFF2025512980000045.tif21170 represents tryptophan, and Y or Tyr represents TIFF2025512980000046.tif19170 represents tyrosine, and V or Val represents TIFF2025512980000047.tif17170 represents valine.

[0035] The term "treatment" includes preventing, slowing, halting or reversing the progression or severity of an existing condition or disease.

[0036] Unless otherwise specified, the term "isomer" is intended to include geometric isomers, cis-trans isomers, stereoisomers, enantiomers, optical isomers, diastereomers and tautomers.

[0037] The compounds of the present invention can exist in specific geometric or stereoisomeric forms. The present invention contemplates all of these compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic and other mixtures thereof, such as enantiomer- or diastereomer-enriched mixtures, and all of these mixtures are within the scope of the present invention. Substituents such as alkyl may have additional asymmetric carbon atoms. All of these isomers and mixtures thereof are within the scope of the present invention.

[0038] Unless otherwise specified, the terms "enantiomers" or "optical isomers" refer to stereoisomers that are mirror images of each other.

[0039] Unless otherwise noted, the term "cis-trans isomers" or "geometric isomers" refers to isomers resulting from the inability to freely rotate about a double bond or a single bond of a ring-forming carbon atom.

[0040] Unless otherwise indicated, the term "diastereomer" means a stereoisomer whose molecules have two or more centers of chirality and which are not mirror-image related to each other.

[0041] Unless otherwise specified, "(+)" denotes dextrorotatory, "(-)" denotes levorotatory, and "(±)" denotes racemic.

[0042] Unless otherwise noted, solid wedge bonds ( TIFF2025512980000048.tif4170) and wedge-dashed bond ( TIFF2025512980000049.tif4170) which shows the absolute configuration of one stereocenter and is represented by a linear solid bond ( TIFF2025512980000050.tif4170) and straight dashed bond ( TIFF2025512980000051.tif5170) shows the relative arrangement of stereocenters, and wavy lines ( TIFF2025512980000052.tif4170) with solid wedge join ( TIFF2025512980000053.tif4170) or wedge-dashed bond ( TIFF2025512980000054.tif4170) or wavy line ( TIFF2025512980000055.tif4170) with straight solid line connection ( TIFF2025512980000056.tif4170) or straight dashed bond ( TIFF2025512980000057.tif5170).

[0043] Unless otherwise specified, the terms "enriched in one isomer," "enriched in one enantiomer," or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100% and is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more, or 99.5% or more, or 99.6% or more, or 99.7% or more, or 99.8% or more, or 99.9% or more.

[0044] Unless otherwise specified, the term "isomeric excess" or "enantiomeric excess" refers to the relative percentage difference between two isomers or two enantiomers. For example, if one isomer or enantiomer is present at 90% and the other isomer or enantiomer at 10%, then the isomeric or enantiomeric excess (ee value) is 80%.

[0045] Optically active (R)- and (S)-isomers, as well as D- and L-isomers, can be prepared using chiral synthesis, chiral reagents, or other conventional techniques. A single enantiomer of a compound of the invention can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary is cleaved to provide the desired pure enantiomer. Alternatively, if the molecule contains a basic (e.g., amino) or acidic (e.g., carboxy) functional group, the diastereomeric salt can be formed with an appropriate optically active acid or base, followed by separation of the diastereomers by conventional methods known in the art, and then recovery to provide the pure enantiomers. Separation of enantiomers and diastereomers is also typically accomplished by chromatographic methods using chiral stationary phases, and optionally in combination with chemical derivatization methods (e.g., carbamate formation from amines).

[0046] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, tritium ( 3 H), iodine-125( 125 I) or C-14( 14 Compounds can be labeled with radioisotopes such as C). For example, hydrogen can be replaced with deuterium to form a deuterated drug, and the bond between deuterium and carbon is stronger than the bond between normal hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have the advantages of reduced toxicity and side effects, increased stability of the drug, enhanced therapeutic effect, and extended biological half-life of the drug. All isotopic constitutional changes of the compounds of the present invention, whether radioactive or not, are all within the scope of the present invention.

[0047] In the case of the linking groups listed, if the linking direction is not indicated, the linking direction is arbitrary, for example When the linking group L in TIFF2025512980000058.tif13170 is -MW-, -MW- links ring A and ring B in the same direction as the reading order from left to right. You can construct TIFF2025512980000059.tif15170 and concatenate rings A and B in the opposite left-to-right reading order: TIFF2025512980000060.tif15170. Combinations of the above linking groups, substituents and / or variants thereof are permissible only if such combinations result in stable compounds.

[0048] Unless otherwise specified, when a group has one or more linkable sites, any one or more of the sites of the group can be linked to other groups via chemical bonds. If the linking mode of the chemical bond is non-directional and there is an H atom at the linkable site, when the chemical bond is linked, the number of H atoms at the site is reduced according to the number of linked chemical bonds to form a group of the corresponding valence. The chemical bond between the above site and other groups is represented by a straight solid line bond ( TIFF2025512980000061.tif5170), Linear dashed line bond ( TIFF2025512980000062.tif5170), or wavy line ( For example, the linear solid bond -OCH3 represents a bond to another group via the oxygen atom of that group, The straight dashed bond in TIFF2025512980000064.tif7170 represents a bond between the nitrogen atom of the group and other groups at both ends. The wavy lines in TIFF2025512980000065.tif13170 represent bonds to other groups via the 1st and 2nd carbon atoms in the phenyl group.

[0049] Unless otherwise specified, "C 1-3 The term "alkyl" is used to denote a linear or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl is C 1-2 and C 2-3 alkyl, etc., and may be monovalent (e.g., methyl), divalent (e.g., methylene) or polyvalent (e.g., methylidyne). 1-3 Examples of alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl) groups, and the like.

[0050] The structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art, and if the present invention relates to the absolute configuration of a compound, its absolute configuration can be confirmed by conventional technical means in the art. For example, in the case of single crystal X-ray diffraction (SXRD), the diffraction intensity data is collected in φ / ω scanning mode using a Bruker D8 venture diffractometer with CuKα radiation as a light source for the cultivated single crystal, and after collecting the relevant data, the crystal structure is further analyzed by a direct method (Shelxs97) to confirm the absolute configuration.

[0051] The compounds of the present invention can be prepared by various synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments in combination with other chemical synthetic methods, and equivalent alternative methods known to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

[0052] The solvents used in the present invention can be obtained commercially.

[0053] In the present invention, the following abbreviations are used: aq stands for water, eq stands for equivalent, DCM stands for dichloromethane, PE stands for petroleum ether, DMSO stands for dimethylsulfoxide, MeOH stands for methanol, BOC stands for tert-butoxycarbonyl, an amine protecting group, rt stands for room temperature, O / N stands for overnight, THF stands for tetrahydrofuran, Boc2O stands for di-tert-butyl dicarbonate, rt-butyl ether, TFA represents trifluoroacetic acid, DIEA represents diisopropylethylamine, DMF represents N,N-dimethylformamide, HBTU represents benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluorophosphate, HOBT represents 1-hydroxybenzotriazole, HOAT represents 1-hydroxy-7-azabenzotriazole, DIC represents N,N'-diisopropylcarbodiimide, DBU represents 1,8-diazabicyclo[5.4.0]undec-7-ene, PhSiH3 represents phenylsilane, and Pd(PPh3)4 represents tetrakis(triphenylphosphine)palladium. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0054] The present invention will be described in detail below with reference to examples, but is not intended to have any adverse limitations to the present invention. In this specification, the present invention is described in detail, but specific embodiments thereof are also disclosed, and it is clear to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. Intermediate A-1 TIFF2025512980000066.tif119170Step 1: Resin fixation 1.1 Weigh out 4 g of chloro(o-chlorophenyl)diphenylmethane (2-CTC Resin (substitution degree S = 1.00 mmol / g)) and 1.54 g of A-1_1 and place them in a reaction column, add DCM (25 mL) thereto, then add 3 mL of N,N-diisopropylethylamine to the reaction column and bubble nitrogen gas for 2 hours, then add 4 mL of MeOH to the reaction column and continue to bubble nitrogen gas for 30 minutes, drain the waste liquid until no liquid comes out, add DMF (50 mL) and wash 5 times, 1 minute each time, and drain the waste liquid until no liquid comes out. 1.2 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. Step 2: Amino acid coupling 2.1 A-1_1 coupling A-1_1 (3.0 eq) was weighed and added to the resin, DIEA (6.00 eq) and 20 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acid was dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. b) The reaction was carried out at 25°C for 20 minutes, ninhydrin was used for detection, and the resin was colorless and transparent. c) The reaction solution was removed and washed with DMF five times for 1 minute each time, and the waste liquid was drained until no more liquid came out. 2.2 A-1_a coupling a) 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no more liquid was produced. Ninhydrin was used for detection, and the resin appeared blue. b) A-1_a (3.0 eq) was weighed and added to the resin, and the reaction column was supplemented with DIEA (3.00 eq) and 20 mL of DMF. Nitrogen gas was passed through the column, and after the amino acid had dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin swelled evenly. c) The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. d) The reaction solution was removed and washed with DMF (50 mL) five times for 1 minute each time, and the waste liquid was drained until no more liquid came out. 2.3 A-1_b coupling a). 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, the waste liquid was drained until no liquid came out, DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. b) Weigh out A-1_b (3.0 eq) and add it to the resin. The reaction column was supplemented with DIEA (6.00 eq) and 20 mL of DMF. Nitrogen gas was passed through the column. After the amino acid had dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin would expand evenly. c) The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. d) The reaction solution was removed and washed with DMF five times for 1 minute each time, and the waste liquid was drained until no more liquid came out. Step 3: Cleavage and drying of crude peptide 3.1 The cutting solution was prepared in the following volumes: TIFF2025512980000067.tif251703.2 60 mL of the prepared cleavage solution was poured into a reactor containing dried peptide resin, the reactor was vented for 20 minutes, filtered, and the filtrate was added to a flask. This procedure was repeated twice, and the cleavage solutions collected in the two procedures were spin-dried to obtain A-1. Intermediate A-2 Intermediate A-2 was obtained by referring to the synthesis of intermediate A-1 in TIFF2025512980000068.tif25170. Step 1. Resin fixation 1.1 4 g of chloro-(o-chlorophenyl)diphenylmethane resin (degree of substitution S=1.00 mmol / g) and 1.54 g of Fmoc-AEEA-OH were weighed and placed in a reaction column, and DCM (25 mL) was added thereto, and then 3 mL of DIEA was added to the reaction column and nitrogen gas was bubbled in for 2 hours, and then 4 mL of MeOH was added to the reaction column and nitrogen gas was bubbled in for 30 minutes, and the waste liquid was discharged until no liquid came out, and DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was discharged until no liquid came out. 1.2 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. Step 2. Coupling of amino acids 2.1 Coupling of Fmoc-AEEA-OH 1. Weigh out Fmoc-AEEA-OH (3.00 eq) and add it to the above resin, then add DIEA (6.00 eq) and 20 mL of DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 2. The reaction was carried out at 25°C for 20 minutes, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF five times for 1 minute each time, and the waste liquid was drained until no more liquid came out. 2.2 Coupling of Fmoc-Glu-OtBu 1. 20% piperidine / DMF (50 mL) was placed in a reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no more liquid was produced. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Glu-OtBu (3.00 eq) and add it to the resin above, then add DIEA (3.00 eq) and 20 mL of DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL) for 1 minute each time, and the waste liquid was drained until no more liquid came out. 2.3 Coupling of 20-(tBu)-eicosanedioic acid 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, the waste liquid was drained until no liquid came out, and DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. 20-(tBu)-eicosanedioic acid (3.00 eq) was weighed and added to the above resin, DIEA (6.00 eq) and 20 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acid was dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin swelled evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF five times for 1 minute each time, and the waste liquid was drained until no more liquid came out. Step 3: Cleavage and drying of crude peptide 3.1 The cutting solution was prepared in the following volumes: TIFF2025512980000069.tif251703.2 This procedure was repeated twice, and the cleavage solution collected twice was spin-dried to obtain A-2. Intermediate A-3 Intermediate A-3 was obtained by referring to the synthesis of intermediate A-1 in TIFF2025512980000070.tif27170. Intermediate A-4 Intermediate A-4 was obtained by referring to the synthesis of intermediate A-1 in TIFF2025512980000071.tif27170.

[0055] Example 1 TIFF2025512980000072.tif52170

[0056] Step 1: 1.34 g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (degree of substitution Sub=0.3 mmol / g) was weighed and placed in a reaction column, DMF (50 mL) was added to the reaction column, nitrogen gas was passed through for 2 hours, and the waste liquid was discharged until no liquid was discharged, DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was discharged until no liquid was discharged.

[0057] Step 2: 20% piperidine / DMF (50 mL) was added to the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no more liquid came out. DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0058] Step 3: Amino acid coupling

[0059] 3.1 Coupling of Fmoc-Ser(tBu)-OH 1. Weigh out Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0060] 3.2 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0061] 3.3 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF five times for 1 minute each time, and the waste liquid was drained until no more liquid came out.

[0062] 3.4 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0063] 3.5 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0064] 3.6 Coupling of Fmoc-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0065] 3.7 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0066] 3.8 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0067] 3.9 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0068] 3.10 Coupling of Fmoc-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0069] 3.11 Coupling of Fmoc-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0070] 3.12 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0071] 3.13 Coupling of Fmoc-Ile-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ile-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0072] 3.14 Coupling of Fmoc-Leu-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Leu-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0073] 3.15 Coupling of Fmoc-Trp(Boc)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Trp(Boc)-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0074] 3.16 Coupling of Fmoc-Lys(Dde)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Lys(Dde)-OH (3.0 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0075] 3.17 Coupling of Fmoc-Val-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Val-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0076] 3.18 Coupling of Fmoc-Phe-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Phe-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0077] 3.19 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0078] 3.20 Coupling of Fmoc-Lys(Alloc)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Lys(Alloc)-OH (2.0 eq) and add it to the above resin, then add HOBT (2.00 eq) and 10 mL DMF to the reaction column, and after the amino acid and HOBT are dissolved, add DIC (2.00 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0079] 3.21 Coupling of Fmoc-Gln(Trt)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gln(Trt)-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0080] 3.22 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0081] 3.23 Coupling of Fmoc-Ile-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ile-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0082] 3.24 Coupling of Fmoc-Lys(Boc)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Lys(Boc)-OH (2.0 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0083] 3.25 Coupling of Fmoc-Asp(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Asp(OtBu)-OH (6.0 eq) and add it to the above resin, add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0084] 3.26 Coupling of Fmoc-Leu-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Leu-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0085] 3.27 Coupling of Fmoc-Aib-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Aib-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 2 hours, and ninhydrin was used for detection; the resin appeared blue. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0086] 3.28 Coupling of Fmoc-Ile-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ile-OH (6.0 eq) and add it to the above resin, then add HOAT (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid and HOAT are dissolved, add DIC (6.00 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 1 hour, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0087] 3.29 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0088] 3.30 Coupling of Fmoc-Tyr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Tyr(tBu)-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0089] 3.31 Coupling of Fmoc-Asp(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Asp(OtBu)-OH (6.0 eq) and add it to the above resin, add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0090] 3.32 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0091] 3.33 Coupling of Fmoc-Thr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Thr(tBu)-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0092] 3.34 Coupling of Fmoc-Phe-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Phe-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0093] 3.35 Coupling of Fmoc-Thr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Thr(tBu)-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0094] 3.36 Coupling of Fmoc-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0095] 3.37 Coupling of Fmoc-Glu(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Glu(OtBu)-OH (6.0 eq) and add it to the resin above. Add HOBT (6.00 eq) and 10 mL DMF to the reaction column. Bubble nitrogen gas. After the amino acid and HOBT were dissolved, add DIC (6.00 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0096] 3.38 Coupling of Fmoc-Aib-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Aib-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out overnight at 25°C, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0097] 3.39 Coupling of Boc-Tyr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Boc-Tyr(tBu)-OH (6.0 eq) and add it to the resin above, then add DIEA (12.0 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0098] 3.40 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (10 mL) were placed in a reaction column, and nitrogen gas was passed through it. Then, Pd(PPh3)4 (0.1 eq) was added and nitrogen gas was passed through it for 20 minutes. The reaction was carried out twice, and the waste liquid was discharged until no more liquid was produced. 2. Wash with DMF 5 times (50 mL each time, 1 min each time) and drain the waste liquid until no more liquid came out.

[0099] 3.41 Coupling of Fmoc-Ida-OH 1. Weigh out Fmoc-Ida-OH (4.0 eq) and add it to the above resin, add DIEA (8.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (3.80 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0100] 3.42 Coupling of intermediate A-1 1. 10% DBU / DMF (50mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out 1.50 eq of intermediate A-1 and add it to the resin. Then add 3.00 eq of DIEA and 10 mL of DMF to the reaction column. After the amino acid was dissolved, add 1.45 eq of HBTU. The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0101] 3.43 De-Dde 1. Add 3% hydrazine / DMF (50mL) to the reaction column, pass nitrogen gas through it for 15 minutes, drain the waste liquid, add DMF (50mL) to wash 5 times, 1 minute each time, drain the waste liquid until no liquid comes out. Ninhydrin was used for detection, and the resin appeared blue.

[0102] 3.44 Amide ring closure 1. DIEA (3.0 eq) was added to the DMF solution of the above resin, and then HATU (1.5 eq) dissolved in DMF was slowly dropped into the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin swelled uniformly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out. 4. The resin was shrunk with MeOH (50 mL) for 3 min each time, drained until no more liquid came out, and the resin was removed, dried, and stored as a spare.

[0103] Step 4. Cleavage and drying of crude peptide

[0104] 4.1 The cleavage solution was prepared in the following volumes: TIFF2025512980000073.tif30170

[0105] 4.2 The dried peptide resin was placed in the prepared cleavage solution, shaken on a shaker for 2.5 hours, filtered, the filtrate was added to 10 times the amount of ice-cold isopropyl ether, centrifuged, and washed with isopropyl ether five times. The crude peptide was obtained by vacuum drying for 2 hours, and purified to obtain polypeptide compound WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1228.6, and the detected value was 1228.7.

[0106] Example 2 Polypeptide WX-002 was obtained by referring to the synthesis of TIFF2025512980000074.tif52170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1243.1, and the detected value was 1242.8.

[0107] Example 3 Polypeptide WX-003 was obtained by referring to the synthesis of TIFF2025512980000075.tif54170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1232.1, and the detected value was 1232.2.

[0108] Example 4 Polypeptide WX-004 was obtained by referring to the synthesis of TIFF2025512980000076.tif54170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1228.6, and the detected value was 1228.2.

[0109] Example 5 TIFF2025512980000077.tif55170

[0110] Step 1: 1.34g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (degree of substitution Sub=0.3mmol / g) was weighed and placed in a reaction column, DMF (50mL) was added to the reaction column, nitrogen gas was passed through for 2 hours, and the waste liquid was discharged until no liquid was discharged, DMF (50mL) was added and washed 5 times, 1 minute each time, and the waste liquid was discharged until no liquid was discharged.

[0111] Step 2: 20% piperidine / DMF (50 mL) was added to the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no more liquid came out. DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0112] Step 3: Amino acid coupling

[0113] 3.1 Coupling of Fmoc-Ser(tBu)-OH 1. Weigh out Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0114] 3.2 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out. 5. Step 3.2 was repeated to complete the coupling of the following amino acids: TIFF2025512980000078.tif234170

[0115] 3.40 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (10 mL) were placed in a reaction column, and nitrogen gas was passed through it. Then, Pd(PPh3)4 (0.1 eq) was added and nitrogen gas was passed through it for 20 minutes. The reaction was carried out twice, and the waste liquid was discharged until no more liquid was produced. 2. Wash with DMF 5 times (50 mL each time, 1 min each time) and drain the waste liquid until no more liquid came out.

[0116] 3.41 Coupling of Fmoc-Ida-OH 1. Weigh out Fmoc-Ida-OH (4.0 eq) and add it to the above resin, add DIEA (8.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (3.80 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0117] 3.42 Coupling of intermediate A-1 1. 10% DBU / DMF (50mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out 1.50 eq of intermediate A-1 and add it to the resin. Then add 3.00 eq of DIEA and 10 mL of DMF to the reaction column. After the amino acid was dissolved, add 1.45 eq of HBTU. The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0118] 3.43 De-Dde 1. 3% hydrazine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 15 minutes, the waste liquid was drained, and DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0119] 3.44 Amide ring closure 1. DIEA (3.0 eq) was added to the DMF solution of the above resin, and then HATU (1.5 eq) dissolved in DMF was slowly dropped into the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin swelled uniformly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out. 4. The resin was shrunk with MeOH (50 mL) for 3 min each time, drained until no more liquid came out, and the resin was removed, dried, and stored as a spare.

[0120] Step 4. Cleavage and drying of crude peptide

[0121] 4.1 The cutting solution was prepared in the following volumes: TIFF2025512980000079.tif30170

[0122] 4.2 The dried peptide resin was placed in the prepared cleavage solution, shaken on a shaker for 2.5 hours, filtered, and the filtrate was added to 10 times the amount of ice-cold isopropyl ether, centrifuged, and washed five times with isopropyl ether. The crude peptide was obtained by vacuum drying for 2 hours, and purified to obtain polypeptide WX-005. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1232.6, and the detected value was 1232.5.

[0123] Example 6 Polypeptide WX-006 was obtained by referring to the synthesis of TIFF2025512980000080.tif49170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1232.4, and the detected value was 1232.4.

[0124] Example 7 Polypeptide WX-007 was obtained by referring to the synthesis of TIFF2025512980000081.tif48170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1218.1, and the detected value was 1218.3.

[0125] Example 8 Polypeptide WX-008 was obtained by referring to the synthesis of TIFF2025512980000082.tif49170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1228.6, and the detected value was 1228.6.

[0126] Example 9 TIFF2025512980000083.tif50170

[0127] Step 1: 1.34g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (degree of substitution Sub=0.3mmol / g) was weighed and placed in a reaction column, DMF (50mL) was added to the reaction column, nitrogen gas was passed through for 2 hours, and the waste liquid was discharged until no liquid was discharged, DMF (50mL) was added and washed 5 times, 1 minute each time, and the waste liquid was discharged until no liquid was discharged.

[0128] Step 2: 20% piperidine / DMF (50 mL) was added to the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no more liquid came out. DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0129] Step 3: Amino acid coupling

[0130] 3.1 Coupling of Fmoc-Ser(tBu)-OH 1. Weigh out Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0131] 3.2 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.0 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out. 5. Step 3.2 was repeated to complete the coupling of the following amino acids: TIFF2025512980000084.tif234170

[0132] 3.40 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (10 mL) were placed in a reaction column, and nitrogen gas was passed through it. Then, Pd(PPh3)4 (0.1 eq) was added and nitrogen gas was passed through it for 20 minutes. The reaction was carried out twice, and the waste liquid was discharged until no more liquid was produced. 2. Wash with DMF 5 times (50 mL each time, 1 min each time) and drain the waste liquid until no more liquid came out.

[0133] 3.41 Coupling of Fmoc-Ida-OH 1. Weigh out Fmoc-Ida-OH (4.0 eq) and add it to the above resin, add DIEA (8.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (3.80 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0134] 3.42 Coupling of intermediate A-1 1. 10% DBU / DMF (50mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out 1.50 eq of intermediate A-1 and add it to the resin. Then add 3.00 eq of DIEA and 10 mL of DMF to the reaction column. After the amino acid was dissolved, add 1.45 eq of HBTU. The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0135] 3.43 De-Dde 1. 3% hydrazine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 15 minutes, the waste liquid was drained, and DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0136] 3.44 Amide ring closure 1. DIEA (3.0 eq) was added to the DMF solution of the above resin, and then HATU (1.5 eq) dissolved in DMF was slowly dropped into the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin swelled uniformly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out. 4. The resin was shrunk with MeOH (50 mL) for 3 min each time, drained until no more liquid came out, and the resin was removed, dried, and stored as a spare.

[0137] Step 4. Cleavage and drying of crude peptide

[0138] 4.1 The cleavage solution was prepared in the following volumes: TIFF2025512980000085.tif30170

[0139] 4.2 The dried peptide resin was placed in the prepared cleavage solution, shaken on a shaker for 2.5 hours, filtered, and the filtrate was added to 10 times the amount of ice-cold isopropyl ether, centrifuged, and washed five times with isopropyl ether. The crude peptide was obtained by vacuum drying for 2 hours, and purified to obtain polypeptide WX-009. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1247.1, and the detected value was 1247.2.

[0140] Example 10 TIFF2025512980000086.tif57170

[0141] Step 1: 1.08g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (degree of substitution Sub=0.37mmol / g) was weighed and placed in a reaction column, DMF (50mL) was added to the reaction column, nitrogen gas was passed through for 2 hours, and the waste liquid was discharged until no liquid was discharged, DMF (50mL) was added and washed 5 times, 1 minute each time, and the waste liquid was discharged until no liquid was discharged.

[0142] Step 2: 20% piperidine / DMF (50 mL) was added to the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no more liquid came out. DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0143] Step 3: Amino acid coupling

[0144] 3.1 Coupling of Fmoc-Ser(tBu)-OH 1. Weigh out Fmoc-Ser(tBu)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0145] 3.2 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0146] 3.3 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0147] 3.4 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0148] 3.5 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0149] 3.6 Coupling of Fmoc-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0150] 3.7 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0151] 3.8 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0152] 3.9 Coupling of Fmoc-Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Pro-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0153] 3.10 Coupling of Fmoc-Gly-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-Gly-OH (3.00 eq) and add it to the above resin, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0154] 3.11 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0155] 3.12 Coupling of Fmoc-Ile-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ile-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0156] 3.13 Coupling of Fmoc-Leu-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Leu-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0157] 3.14 Coupling of Fmoc-Trp(Boc)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Trp(Boc)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0158] 3.15 Coupling of Fmoc-Glu(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Glu(OtBu)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0159] 3.16 Coupling of Fmoc-Val-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Val-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0160] 3.17 Coupling of Fmoc-Phe-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Phe-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0161] 3.18 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0162] 3.19 Coupling of Fmoc-Lys(Alloc)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Lys(Alloc)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0163] 3.20 Coupling of Fmoc-Gln(Trt)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gln(Trt)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0164] 3.21 Coupling of Fmoc-Ala-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ala-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0165] 3.22 Coupling of Fmoc-Lys(Dde)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Lys(Dde)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0166] 3.23 Coupling of Fmoc-Lys(Boc)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Lys(Boc)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0167] 3.24 Coupling of Fmoc-Asp(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Asp(OtBu)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0168] 3.25 Coupling of Fmoc-Leu-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Leu-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0169] 3.26 Coupling of Fmoc-Aib-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Aib-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 2 hours, and ninhydrin was used for detection; the resin appeared blue. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0170] 3.27 Coupling of Fmoc-Ile-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ile-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 1 hour, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0171] 3.28 Coupling of Fmoc-Ser(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Ser(tBu)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0172] 3.29 Coupling of Fmoc-Tyr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Tyr(tBu)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0173] 3.30 Coupling of Fmoc-Asp(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Asp(OtBu)-OH (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0174] 3.31 Coupling of Fmoc-Thr(tBu)-SerPsi(Me,Me)Pro-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Thr(tBu)-SerPsi(Me,Me)Pro-OH (2.00 eq) and add it to the above resin, then add DIEA (4.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (1.90 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0175] 3.32 Coupling of Fmoc-Phe-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Phe-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0176] 3.33 Coupling of Fmoc-Thr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Thr(tBu)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0177] 3.34 Coupling of Fmoc-Gly-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Gly-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0178] 3.35 Coupling of Fmoc-Glu(OtBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Glu(OtBu)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0179] 3.36 Coupling of Fmoc-Aib-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out Fmoc-Aib-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out overnight at 25°C, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0180] 3.37 Coupling of Boc-Tyr(tBu)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weigh out Boc-Tyr(tBu)-OH (3.00 eq) and add it to the resin above, then add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would swell evenly. 3. The reaction was carried out at 25°C for 0.5 hours, chloranil was used for detection, and the resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0181] 3.38 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (20 mL) were placed in a reaction column, and nitrogen gas was passed through it. Then, Pd(PPh3)4 (0.10 eq) was added and nitrogen gas was passed through it for 20 minutes. The reaction was carried out twice, and the waste liquid was discharged until no more liquid was produced. 2. Wash with DMF 5 times (50 mL each time, 1 min each time) and drain the waste liquid until no more liquid came out.

[0182] 3.39 Coupling of Fmoc-Glu-OAll 1. Weigh out Fmoc-Glu-OAll (3.00 eq) and add it to the above resin, add DIEA (6.00 eq) and 10 mL DMF to the reaction column, and then pass nitrogen gas through the column. After the amino acid is dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0183] 3.40 Coupling of intermediate A-1 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was bubbled through for 20 minutes, and the waste liquid was drained until no liquid came out. DMF (50 mL) was added and washed 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh out intermediate A-1 (1.50 eq) and add it to the above resin, then add DIEA (3.00 eq) and 10 mL DMF to the reaction column, and after the amino acid is dissolved, add HATU (1.45 eq). The nitrogen gas was adjusted so that the resin would expand evenly. 3. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out.

[0184] 3.41 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (20 mL) were placed in a reaction column, and nitrogen gas was passed through it. Then, Pd(PPh3)4 (0.10 eq) was added and nitrogen gas was passed through it for 20 minutes. The reaction was carried out twice, and the waste liquid was discharged until no more liquid was produced. 2. Wash with DMF 5 times (50 mL each time, 1 min each time) and drain the waste liquid until no more liquid came out.

[0185] 3.42 De-Dde 1. 3% hydrazine hydrate / DMF (50 mL) was placed in a reaction column, nitrogen gas was bubbled through for 15 minutes, the waste liquid was drained, and DMF (50 mL) was added to wash 5 times, 1 minute each time, and the waste liquid was drained until no liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0186] 3.43 Amide ring closure 1. DIEA (3.00 eq) was added to the DMF solution of the above resin, and then HATU (1.50 eq) dissolved in DMF was slowly dropped into the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin swelled uniformly. 2. The reaction was carried out at 25°C for 0.5 hours, and ninhydrin was used for detection. The resin was colorless and transparent. 3. The reaction solution was removed and washed with DMF 5 times (50 mL each time, 1 minute each time), and the waste liquid was drained until no more liquid came out. 4. The resin was shrunk with MeOH (50 mL) for 3 min each time, drained until no more liquid came out, and the resin was removed, dried, and stored as a spare.

[0187] Step 4. Cleavage and drying of crude peptide

[0188] 4.1 The cleavage solution was prepared in the following volumes: TIFF2025512980000087.tif30170

[0189] 4.2 The dried peptide resin was placed in the prepared cleavage solution, shaken on a shaker for 2.5 hours, filtered, and the filtrate was added to 10 times the amount of ice-cold isopropyl ether, centrifuged, and washed five times with isopropyl ether. The crude peptide was obtained by vacuum drying for 2 hours and purified. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value was 1236.1, and the detected value was 1236.0.

[0190] Example 11 Polypeptide WX-011 was obtained by referring to the synthesis of TIFF2025512980000088.tif62170WX-010. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1236.1, and the detected value was 1236.0.

[0191] Example 12 TIFF2025512980000089.tif55170 Using intermediate A-2, polypeptide WX-012 was obtained by referring to the synthesis of WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1225.6, and the detected value was 1225.7.

[0192] Example 13 TIFF2025512980000090.tif59170 Using intermediate A-3, polypeptide WX-013 was obtained by referring to the synthesis of WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1264.9, and the detected value was 1264.6.

[0193] Example 14 Polypeptide WX-014 was obtained using intermediate A-4 by referring to the synthesis of TIFF2025512980000091.tif54170WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, and the calculated value [M+4H] / 4 was 1257.9, and the detected value was 1257.8.

[0194] Biological Test Data

[0195] Test Example 1: In vitro GLP-1R / GIPR agonist activity test

[0196] A:Main materials: 1) Cell line This cell line was constructed by Wuxi APPTEC (Shanghai) Co., Ltd. For details, see Table 1 below. TIFF2025512980000092.tif241702) Reagents and Consumables TIFF2025512980000093.tif791703) Equipment TIFF2025512980000094.tif44170

[0197] B. Method 1) Experimental materials Experimental buffer TIFF2025512980000095.tif60170 Preparation of assay reagents TIFF2025512980000096.tif331702) Experimental method a) Preparation of compound plates: Test compounds were diluted 4-fold in 10-point runs from an initial concentration of 30 μM, performed by Bravo. b) Transport of compounds: 1) 100 nL of compound was transferred to an OptiPlate-384 plate using the Echo. 2) The OptiPlate-384 plate was centrifuged at 1000 rpm for 5 seconds. c) Preparation of cell suspension 1) One of the cryopreserved vials of GLP-1R / GIPR cells was rapidly thawed in 37°C warm water. 2) The cell suspension was transferred to a 15 mL centrifuge tube and gently washed with 10 mL of HBSS. 3) The tube was centrifuged at room temperature at 1000 rpm for 1 minute. 4) The supernatant was discarded. 5) The cells at the bottom were gently dispersed, then gently washed with 10 mL HBSS, the cells were centrifuged to sediment, and finally the cells were resuspended in the experiment buffer. 6) Cell density and viability were measured using Vi-cell. 7) GLP-1R / GIPR cells at a concentration of 2.0*10 5 The solution was diluted with the experimental buffer to give a concentration of 1 / mL. 8) 100 nL of the diluted cell suspension was transferred to an OptiPlate-384 plate. 9) Incubate at room temperature for 30 minutes. d) Addition of Assay Reagents: 1) 10 μL of 800 nM gradient diluted cAMP standard solution was added to an empty well of an OptiPlate-384 plate. 2) 10 μL of cAMP assay reagent was added. 3) The OptiPlate-384 plate was covered with TopSeal-A film and incubated at room temperature for 60 minutes. 4) TopSeal-A was removed and the plate was read using EnVision.

[0198] C. Experimental Results The experimental results are shown in Table 6. TIFF2025512980000097.tif99170 Conclusion: The compounds of the present invention show very strong agonist activity against GLP-1R / GIPR.

[0199] Test Example 2: Evaluation of the pharmacokinetic profile of the compound in rats A. Purpose of the experiment The purpose was to study the pharmacokinetic profile of the compound in SD rats. B. Experimental Procedure The pharmacokinetic properties of the compounds in rodents after subcutaneous injection were tested using standard protocols. In the experiments, the candidate compounds were formulated in a clear solution and administered to rats by a single subcutaneous injection (SC, 0.048 mpk). The injection vehicle was citrate buffer (20 mM, pH=7). Whole blood was collected and prepared as plasma, the drug concentrations were analyzed by LC-MS / MS method, and the pharmacokinetic parameters were calculated with Phoenix WinNonlin software. C. Experimental Results The experimental results are shown in Table 7. TIFF2025512980000098.tif33170 Conclusion: The compounds of the present invention have excellent pharmacokinetic properties in rats.

[0200] Test Example 3: Evaluation of the pharmacokinetic profile of the compound in mice A. Purpose of the experiment The purpose was to study the pharmacokinetic profile of the compound in C57BL / 6 mice. B. Experimental Procedure The pharmacokinetic properties of the compounds in rodents after subcutaneous injection were tested using standard protocols. In the experiments, the candidate compounds were formulated in a clear solution and administered to rats by subcutaneous injection (SC, 0.048mpk). The subcutaneous injection vehicle was citrate buffer (20mM, pH=7). Whole blood was collected and prepared as plasma, the drug concentrations were analyzed by LC-MS / MS method, and the pharmacokinetic parameters were calculated with Phoenix WinNonlin software. C. Experimental Results The experimental results are shown in Table 8. TIFF2025512980000099.tif33170 Conclusion: The compounds of the present invention have excellent pharmacokinetic properties in mice.

[0201] Test Example 4: Evaluation of the pharmacokinetic profile of a compound in cynomolgus monkeys A. Purpose of the experiment The purpose was to study the pharmacokinetic profile of the compound in cynomolgus monkeys. B. Experimental Procedure The pharmacokinetic properties of compounds in mammals after intravenous and subcutaneous administration were tested using standard protocols. In the experiments, candidate compounds were formulated in clear solutions and administered to cynomolgus monkeys by a single subcutaneous injection (SC, 0.02 mpk). The subcutaneous injection vehicle was citrate buffer (20 mM, pH=7). Whole blood was collected and prepared as plasma, the drug concentrations were analyzed by LC-MS / MS method, and the pharmacokinetic parameters were calculated with Phoenix WinNonlin software. C. Experimental Results The experimental results are shown in Table 9. TIFF2025512980000100.tif33170 Conclusion: Compounds of the invention have excellent pharmacokinetic properties in monkeys.

[0202] Test Example 5: Plasma stability test (PLS) A. Purpose of the experiment The purpose was to study the stability of the test compound in the plasma of normal mice. B. Experimental Procedure 1. Before the experiment, the clotted frozen plasma was thawed in a water bath at 37° C. The plasma was centrifuged at 4000 rpm for 5 min, and blood clots, if any, were removed, and the pH was adjusted to 7.4±0.1. 2. Preparation of test compound solution: Dilute with DMSO to prepare a 100 μM solution. 3. 98 μL of blank control plasma was added to 2 μL of a test compound solution (100 μM) so that the final concentration of the mixture became 2 μM, and this was incubated under water bath conditions at 37°C. 4. At each time point (0, 10, 30, 60 and 120 min), 100 μL H3PO4 solution and 800 μL stop solution (200 ng / mL tolbutamide and 200 ng / mL labetalol in 100% methanol) were added respectively to precipitate proteins and mixed thoroughly. 5. The samples were centrifuged at 4000 rpm for 20 minutes, and 100 μL of the supernatant was taken from each well for LC-MS / MS analysis. C. Experimental Results The experimental results are shown in Table 10. TIFF2025512980000101.tif18170 Conclusion: The compounds of the present invention have excellent plasma stability.

[0203] Test Example 6: Plasma Protein Binding Test (PPB) A. Purpose of the experiment The purpose was to study the binding ability of the test compounds to human / mouse plasma albumin. B. Experimental Procedure 1. Preparation of matrix (preparation of blank matrix): On the day of the experiment, thaw the plasma in cold water and centrifuge at 3220 rpm for 5 minutes to remove any clots. The pH of the obtained plasma was measured and adjusted to 7.4±0.1 with 1% phosphoric acid or 1N sodium hydroxide as necessary. 2. Dilution process of test compounds: Test compounds were dissolved in dimethyl sulfoxide (DMSO) to prepare stock solutions with concentrations of 10 mM and 2 mM, respectively. 2 μL of stock solution (2 mM) was diluted with 98 μL of DMSO to prepare a standard solution of 40 μM. 10 μL of stock solution was diluted with 240 μL of DMSO to prepare a standard solution of 400 μM of the control compound. The compound standard solution (5 μL) was mixed evenly with the blank matrix (995 μL) in a ratio of 1:200 to prepare the loading matrix. 3. Analysis process 3.1 Equal amounts of 30 μL loading matrix (n=2) were transferred to a sample collection plate to prepare test time 0 (T0) samples for residue measurement. Samples were immediately combined with the corresponding blank buffer to a final volume of 60 μL, such that the volume ratio of plasma (loading matrix) to buffer was 1:1 in each well. Then, 60 μL of 4% H3PO4 in H2O and 480 μL of stop solution containing the internal standard were added to the T0 samples of the test compounds, respectively. They were then stored at 2-8 °C for further processing along with the other samples. 3.2 The remaining plasma samples were pre-incubated for 30 min in a carbon dioxide incubator at 37 ± 1°C. Protein-free samples (F samples) were prepared and transferred together with the matrix-loaded samples (230 μL) into polycarbonate tubes (n = 2) and ultracentrifuged at 37°C and 155,000 × g (35,000 rpm) for 4 h. 3.3 To prepare T samples (test samples), one additional matrix-containing sample was transferred to a single 96-well plate (sample incubation plate) and incubated at 37° C. for 4 hours. 3.4 After centrifugation was completed, 30 μL of protein-free sample (F sample) and 30 μL of T sample were taken from the second layer of the supernatant (below the upper layer) and transferred to a new sample collection plate. Each sample was mixed with the corresponding blank buffer or matrix (blank matrix) to a final volume of 60 μL. The volume ratio of matrix (blank matrix):buffer was 1:1. 60 μL of 4% H3PO4 aqueous solution and 480 μL of stop solution (containing internal standard) were added to all samples. The mixtures were centrifuged at 4000 rpm for 20 min, and 100 μL of supernatant was taken from each sample for LC-MS / MS analysis. C. Experimental Results The experimental results are shown in Table 11. TIFF2025512980000102.tif23170Note: NA indicates that plasma protein binding capacity was so high that free drug was not detectable at normal plasma protein concentrations. Conclusion: The compounds of the present invention show extremely high plasma protein binding capacity.

[0204] Test Example 7: In vivo efficacy test in an obese mouse model A. Purpose of the experiment The weight loss effects of a series of polypeptides of the present invention were investigated in obese animal models and compared with the commercially available equivalents Tirzepatide and Semaglutide. B. Experimental Procedure 1. Animal modeling The obese mouse model was obtained by high-fat diet induction according to the reference (MOLECULAR METABOLISM, 18 (2018), P3~14). Regarding the specific experimental procedure, the subject mice were fed a high-fat diet (Research Diet D12492) for 13 weeks. The weight of the normal control group and the model animals increased steadily, but the weight of the model group increased significantly more than the control group, and the intake was also significantly higher than the control group, suggesting that the modeling was successful. 2. Administration The test animals were divided into groups, and according to the experimental protocol, appropriate amounts of test samples were added to 20 mM sodium citrate buffer, vortexed at room temperature to create clear solutions of different concentrations, and then frozen and stored. During the experiment, the animals in the model group and treatment group were administered the drug via subcutaneous injection in the outer thigh, once every three days (Q3d), for a total of 22 days. The details are as follows: TIFF2025512980000103.tif47170C Results and Analysis During the experiment, all test mice were in good health and mental condition. The animals in the treatment groups showed different degrees of food intake suppression due to the effects of the drug, but no deaths occurred. After the completion of all treatment cycles, the test mice were weighed and the results are shown below. TIFF2025512980000104.tif41170 Conclusion: Both WX001 and WX005 showed good weight loss effects in obese mouse models, where WX005 showed better efficacy and was superior to tirzepatide and semaglutide. In repeated experiments, the test drug showed substantially consistent effects across corresponding groups, indicating the existence of objective efficacy benefits. Moreover, the beneficial effect of weight loss was also observed with other polypeptide compounds of the present invention.

[0205] Test Example 8: In vivo efficacy test in diabetic mouse model A. Purpose of the experiment The efficacy of a series of polypeptides of the present invention was examined in diabetic animal models and compared with the commercially available equivalent Tirzepatide. According to a report in the literature (MOLECULAR METABOLISM, 18 (2018), P3-14), the blood glucose lowering effect of Tirzepatide is much better than that of Semaglutide, so Semaglutide was not included in this experiment for comparison. B. Experimental Procedure 1. Selection of Animal Model The db / db (Lepr leptin receptor-deficient) mouse has typical clinical symptoms of diabetes, such as extreme obesity, polyphagia, dry mouth, and polyuria, and is an ideal animal model of type 2 diabetes. Specific experimental procedures: db / db test mice weighing more than 30g and fasting blood glucose levels more than 15mmol / L were screened for inclusion in the formal study. 2. Administration The test animals were divided into groups, and according to the experimental protocol, an appropriate amount of the test sample was added to 20 mM sodium citrate buffer, vortexed to a clear solution at room temperature, and then frozen and stored. During the experiment, the animals in the normal control group, model group and treatment group were administered the drug via subcutaneous injection in the outer thigh, once a day (Qd), for a total of 4 weeks. The details are as follows: TIFF2025512980000105.tif41170C Results and Analysis During the experiment, all the test mice were in good health and mental condition. The animals in the treatment group showed different degrees of food intake suppression due to the influence of the drug, but none died. During the experiment, the fasting blood glucose and glycosylated hemoglobin change rates of the test mice were dynamically monitored, where the fasting blood glucose levels are shown below. TIFF2025512980000106.tif47170The results of the rate of change in glycated hemoglobin values ​​were as follows. TIFF2025512980000107.tif47170Note: "-" indicates a decrease. Conclusion: Both WX005 and WX009 show good blood glucose lowering effects in mouse diabetes models, specifically, WX005 and WX009 show similar effects in lowering the glycated hemoglobin levels of the test animals and are superior to tirzepatide, WX005 is slightly superior to WX009 in lowering the fasting blood glucose levels of the test animals, and both are superior to tirzepatide. Overall, WX005 and WX009 are considered to have significantly superior blood glucose lowering effects than tirzepatide. In repeated experiments, the test drug showed substantially consistent effects across corresponding groups, indicating the existence of objective efficacy benefits. Moreover, efficacy benefits regarding glucose metabolism in animals have also been observed with other polypeptide compounds of the invention.

Claims

1. The sequence is of formula (II), i.e. YAibEGT FTSDY SIAibLD KIAQK AFVKW LIAGG PSSGA PPPS 0 (II) is shown in As shown below: 1) The amino acid at position i is replaced with lysine, and then the amino group on the lysine is combined with the amino group on the lysine side chain at position i+3 to form a compound of the formula or the amino groups on the lysine side chain at positions j and j+4 are linked to groups represented by the formula wherein i is 17 and j is 20; and 2) an additional 0 to 2 amino acids of the polypeptide of formula (II) are substituted; Where: The structure of Aib is of the formula is a group represented by S ０ is the formula is selected from the group represented by X is of the formula wherein "*" represents X １ represents the position connected to X １ represents a single bond, -C(=O)-, -O-C(=O)-, and -N(R １ )-C(=O)-; R １ is H and C １－３ alkyl, X ２ is the formula is selected from the group represented by m is selected from 2, 3 and 4; n is selected from 15, 16, 17, 18 and 19; p comprises a modification selected from 1 and 2 The pharmaceutical use of a polypeptide in the preparation of a medicament for treating and / or preventing diabetes, obesity and related disorders.

2. The sequence of the polypeptide is shown in formula (P): YAibEGT FTSDY SIAibLD KKAQK AFVKW LIAGG PSSGA PPPS 0 (P) As shown below: 1) The amino groups on the lysine side chains at positions 17 and 20 are of the formula and 2) 0 to 2 amino acids of the polypeptide are substituted; Where: The structure of Aib is of the formula is a group represented by S ０ is the formula is selected from the group represented by X is of the formula wherein "*" represents X １ represents the position where the X １ represents a single bond, -C(=O)-, -O-C(=O)-, and -N(R １ )-C(=O)-; R １ is H and C １－３ alkyl, X ２ is the formula is selected from the group represented by m is selected from 2, 3 and 4; n is selected from 15, 16, 17, 18 and 19; p is selected from 1 and 2 Includes modifications 2. A pharmaceutical use of the polypeptide of claim 1.

3. 0 to 2 amino acids at positions 21, 23, or 24 of said polypeptide are substituted 3. Pharmaceutical use of a polypeptide according to claim 1 or 2.

4. The polypeptides have sequences represented by formulae (II-1), (II-2), (II-3), (II-4), (III-6), (IV-7), (IV-8), (IV-9) and (IV-10), YAibEGT FTSDY SIAibLD KIAQK AFVKW LIAGG PSSGA PPPS-NH 2 (II-1) YAibEGT FTSDY SIAibLD KIAQK EFVKW LIAGG PSSGA PPPS-NH 2 (II-2) YAibEGT FTSDY SIAibLD KIAQK AFIKW LIAGG PSSGA PPPS-NH 2 (II-3) YAibEGT FTSDY SIAibLD KIAQK AFVKW LLAGG PSSGA PPPS-NH 2 (II-4) YAibEGT FTSDY SIAibLD KKAQK AFVEW LIAGG PSSGA PPPS-NH 2 (III-6) YAibEGT FTSDY SIAibLD KKAQK AFVQW LIAGG PSSGA PPPS-NH 2 (IV-7) YAibEGT FTSDY SIAibLD KKAQK AFVAW LIAGG PSSGA PPPS-NH 2 (IV-8) YAibEGT FTSDY SIAibLD KKAQK AFVIW LIAGG PSSGA PPPS-NH 2 (IV-9) YAibEGT FTSDY SIAibLD KKAQK EFVEW LIAGG PSSGA PPPS-NH 2 (IV-10) As shown below: The amino groups on the lysine side chains at positions 17 and 20 or the amino groups on the lysine side chains at positions 20 and 24 are represented by the formula is linked to a group represented by Where: Aib, X, X １ and X ２ includes the modification as defined in claim 1 2. A pharmaceutical use of the polypeptide of claim 1.

5. In the polypeptide, R １ is H 5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

6. In the polypeptide, X １ is selected from the group consisting of a single bond, -C(=O)-, -O-C(=O)-, and -NH-C(=O)-.

5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

7. In the above polypeptide, m is 2.

5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

8. In the above polypeptide, n is selected from 15 and 17.

5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

9. In the above polypeptide, p is 2.

5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

10. In the polypeptide, X ２ teeth, formula is selected from the group consisting of 5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

11. X ２ is the formula is selected from the group consisting of The polypeptide of claim 10.

12. The amino groups on the lysine side chain at positions i and i+3 or the amino groups on the lysine side chain at positions j and j+4 are represented by the formula and linking to a group represented by the formula Form a group represented by A polypeptide according to any one of claims 1 to 4.

13. The amino groups on the lysine side chain at positions i and i+3 or the amino groups on the lysine side chain at positions j and j+4 are represented by the formula and linking to a group represented by the formula Form a group represented by A polypeptide according to any one of claims 1 to 4.

14. In the polypeptide, The structural unit represented by formula is selected from the group consisting of 5. A pharmaceutical use of a polypeptide according to any one of claims 1 to 4.

15. In the polypeptide, The structural unit represented by formula is selected from the group consisting of 15. A pharmaceutical use of the polypeptide of claim 14.

16. A pharmaceutical use of a polypeptide having a sequence represented by the following formula in the preparation of a medicament for treating and / or preventing diabetes, obesity and related diseases.

17. A pharmaceutical composition for treating and / or preventing diabetes, obesity and related diseases, comprising a therapeutically effective amount of the compound according to any one of claims 1 to 16 or a pharma- ceutical acceptable salt thereof as an active ingredient, and a pharma- ceutical acceptable carrier.

18. The diabetes-related diseases include diabetic cardiovascular disease, diabetic cerebrovascular disease, retinopathy, nephropathy, cataract, coronary artery disease, diabetic neuropathy, diabetic foot, peripheral neuropathy, etc., and the obesity-related diseases include liver disease, hyperlipidemia, hypertension, etc. A pharmaceutical use of a polypeptide or a pharma- ceutical acceptable salt thereof according to any one of claims 1 to 16 in the preparation of a pharmaceutical product for treating and / or preventing diabetes, obesity and related diseases, or a pharmaceutical composition for treating and / or preventing diabetes, obesity and related diseases according to claim 17.

19. The pharmaceutical agent for treating and / or preventing diabetes, obesity and related diseases is administered once every 3 days, once every 4 days, once every week or once every 2 weeks. A pharmaceutical use of a polypeptide or a pharma- ceutical acceptable salt thereof according to any one of claims 1 to 16 in the preparation of a pharmaceutical product for treating and / or preventing diabetes, obesity and related diseases, or a pharmaceutical composition for treating and / or preventing diabetes, obesity and related diseases according to claim 17.

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