Medicinal use of polypeptides in the preparation of pharmaceuticals for the treatment and / or prevention of diabetes, obesity and related diseases.

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 pharmaceutically acceptable use of a polypeptide whose sequence is represented by formula (II) in the preparation of pharmaceuticals for treating and / or preventing diabetes, obesity and related diseases. [Background technology]

[0002] The global prevalence of non-alcoholic fatty liver disease (NAFLD) is high at 25%, with non-alcoholic steatohepatitis (NASH) accounting for approximately 3% to 8% of that population. Some NASH patients progress to hepatic sclerosis and liver cancer, and it is now one of the leading causes of end-stage liver disease and liver transplantation. The pathogenesis of NASH is complex, and there are currently no FDA-approved treatments for the disease. Several preclinical studies have shown that glucose-dependent insulinotropic polypeptide (GIP) / glucagon-like peptide-1 (GLP-1) dual agonists may be useful in treating NASH. Eli Lilly's clinical trial of Tirzepatide, a GLP-1 / GIP dual agonist currently under investigation, has shown that it improves NASH-related markers such as transaminases, demonstrating its potential as a treatment for NASH.

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

[0004] Furthermore, during the course of research, it was discovered that GLP-1 / GIP dual agonists have excellent effects on regulating glucose and lipid metabolism in the body. Compounds targeting this substance have the potential to be developed as preventive and / or therapeutic agents for various specific diseases involving glucose metabolism abnormalities and / or lipid metabolism abnormalities, such as diabetes, obesity, and their associated complications (related diseases). [Overview of the project]

[0005] The present invention relates to an arrangement of formula (II), i.e. YAibEGT FTSDY SIAibLD KIAQK AFVKW LIAGG PSSGA PPPS0 (II) As shown, As follows, 1) The amino group on the lysine side chain at positions i and i+3 or the amino group on the lysine side chain at positions j and j+4 It is linked to TIFF0007886963000001.tif10170, where i is 17 (i.e., when i is 17, first the isoleucine at position 17 is replaced with lysine, 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 linked to X respectively, and the amino group (concatenated via TIFF0007886963000002.tif10170), or where j is 20 and 2) In the series of polypeptides represented by formula (II), an additional 0 to 2 amino acids are substituted, Here, The structure of Aib is, The filename is TIFF0007886963000003.tif15170, S0 has a structure The serine (S,(S)-2-amino-3-hydroxypropionic acid) with the structure of TIFF0007886963000004.tif17170 is Selected from (S)-2-amino-3-hydroxypropionamide (i.e., the amino acid at position 39 (serine) is optionally amidated to a primary amide at the C-terminus), TIFF0007886963000005.tif17170 X is Selected from TIFF0007886963000006.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 Selected from alkyl groups, X2 is Selected from TIFF0007886963000007.tif38170, m is selected from 2, 3, and 4. n is selected from 15, 16, 17, 18, and 19. p includes modifications selected from 1 and 2. This invention provides for the pharmaceutically acceptable use of polypeptides in the preparation of pharmaceuticals for the treatment and / or prevention of diabetes, obesity, and related diseases.

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

[0007] The present invention relates to an arrangement of formula (P), i.e., YAibEGT FTSDY SIAibLD KKAQK AFVKW LIAGG PSSGA PPPS0 (P) As shown, As follows, 1) The amino groups on the lysine side chains at positions 17 and 20 It is linked to TIFF0007886963000008.tif10170, and 2) In polypeptides whose sequence is represented by formula (P), 0 to 2 amino acids are substituted. Here, The structure of Aib is The filename is TIFF0007886963000009.tif15170, S0 has a structure TIFF0007886963000010.tif18170 is serine (S,(S)-2-amino-3-hydroxypropionic acid), and its structure is Selected from (S)-2-amino-3-hydroxypropionamide (i.e., the amino acid at position 39 (serine) is optionally amidated to a primary amide at the C-terminus), TIFF0007886963000011.tif18170 X is Selected from TIFF0007886963000012.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 Selected from alkyl groups, X2 is Selected from TIFF0007886963000013.tif38170, m is selected from 2, 3 and 4, n is selected from 15, 16, 17, 18 and 19, p includes modifications selected from 1 and 2 Provided is the pharmaceutical use of a polypeptide in the preparation of a medicament for treating and / or preventing diabetes, obesity and related diseases.

[0008] The polypeptide sequence represented by the formula (P) is as shown by Seq ID NO.2.

[0009] In some aspects of the present invention, 0 to 2 amino acids at positions 21, 23 or 24 of the above polypeptide are substituted, and the remaining variables are as defined in the present invention.

[0010] In some aspects of the present invention, the above polypeptide has a sequence selected from the sequences represented by the 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 the formulas (II-1), (II-2), (II-3), (II-4), (III-6), (IV-7), (IV-8), (IV-9) and (IV-10) are respectively represented by Seq ID NO.3 to 11. 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 follows, 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 It is linked to TIFF0007886963000014.tif10170, Here, the serine at position 39 is amidated to the primary amide at the C-terminus, where Aib, X, X1, and X2 are as defined in this invention.

[0011] In some aspects of the present invention, R1 is selected from H, and the remaining variables are as defined in the present invention.

[0012] In some aspects of the present invention, X1 is selected from a single bond, -C(=O)-, -OC(=O)-, and -NH-C(=O)-, and the remaining variables are as defined in the present invention.

[0013] In some aspects of the present invention, the above m is selected from 2, and the remaining variables are as defined in the present invention.

[0014] In some aspects of the present invention, n is selected from 15 and 17, and the remaining variables are as defined in the present invention.

[0015] In some aspects of the present invention, the above X2 is Selected from TIFF0007886963000015.tif76170, the remaining variables are as defined in this invention.

[0016] In some aspects of the present invention, the above X2 is Selected from TIFF0007886963000016.tif75170, the remaining variables are as defined in this invention.

[0017] In some aspects of the present 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 Concatenate to TIFF0007886963000017.tif10170, The file TIFF0007886963000018.tif93170 is formed, and the remaining variables are as defined in this invention.

[0018] In some aspects of the present 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 Concatenate to TIFF0007886963000019.tif10170, The file TIFF0007886963000020.tif92170 is formed, and the remaining variables are as defined in this invention.

[0019] In some aspects of the present invention, TIFF0007886963000021.tif10170 is, Selected from TIFF0007886963000022.tif176170, the remaining variables are as defined in this invention.

[0020] In some aspects of the present invention, TIFF0007886963000023.tif10170 is, Selected from TIFF0007886963000024.tif122170, the remaining variables are as defined in this invention. The present invention also has several aspects that can be derived from any combination of the above variables.

[0021] The present invention further provides the pharmaceutically acceptable use of polypeptides selected from polypeptides represented by the following formula in the preparation of pharmaceuticals for treating and / or preventing diabetes, obesity and related diseases. TIFF0007886963000025.tif234170TIFF0007886963000026.tif240170TIFF0007886963000027.tif245170

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

[0023] Diabetes mellitus is a chronic disease accompanied by many pathological symptoms. It is a metabolic and endocrine disorder caused by an absolute deficiency of insulin or a decrease in the biological effect of insulin, and is one of the specific indications for lipid metabolism disorders, circulatory disorders, and / or glucose metabolism disorders in the body. Diabetes mellitus can easily lead to various complications (related diseases) such as diabetic cardiovascular disease, diabetic cerebrovascular disease, retinopathy, nephropathy, cataracts, coronary artery disease, diabetic neuropathy, diabetic foot, and peripheral neuropathy. Among these, diabetic cardiovascular complications include microvascular disease of the heart and major blood vessels, cardiomyopathy, and cardiac autonomic neuropathy, while diabetic cerebrovascular diseases include cerebral arteriosclerosis, ischemic cerebrovascular disease, cerebral hemorrhage, and cerebral atrophy.

[0024] Obesity is another common metabolic syndrome caused by a combination of factors including unhealthy eating habits, lack of exercise, emotional factors, and genetic factors, and is one of the specific indications for abnormal lipid metabolism, circulatory disorders, and / or glucose metabolism in the body. On the other hand, obesity increases the risk of developing related diseases such as liver disease, hyperlipidemia, diabetes, and hypertension.

[0025] Diabetes, obesity, and related diseases have a significant impact on human health and socioeconomics.

[0026] In some aspects of the present invention, the use of the above polypeptide compounds or pharmaceutically acceptable salts thereof or the above pharmaceutical compositions in the preparation of pharmaceuticals for the treatment of diabetes and / or obesity is provided.

[0027] In some aspects of the present invention, the present invention provides the above polypeptide compound or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition, which is administered at a frequency of once every three days, once every four days, once a week, or once every two weeks. Technical effects

[0028] The polypeptide of the present invention has very strong agonist 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. Definition and description

[0029] Unless otherwise specified, the following terms and phrases used herein shall have the meanings set forth below. Unless otherwise specified, certain terms or phrases should not be considered ambiguous or unclear, but rather understood in their general sense. Where a trade name is mentioned herein, it is intended to refer to the corresponding product or its active ingredient.

[0030] As used herein, the term “pharmaceutically acceptable” means, within the bounds of reliable medical judgment, a compound, material, composition, and / or dosage form that is suitable for use in contact with human and animal tissues in a reasonable benefit-to-risk ratio, without excessive toxicity, irritation, allergic reactions, or other problems or complications.

[0031] The term "pharmaceutically acceptable salt" means a salt of the polypeptide compound of the present invention prepared from a compound having certain substituents found in the present invention and a relatively non-toxic acid or base. If 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 a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium salts, potassium salts, calcium salts, ammonium salts, organic amine or magnesium salts or similar salts. If 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 a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, which include hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, and phosphorous acid; and organic acid salts, which include acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanodioic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and similar acids; and also salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Some specific compounds of the present invention contain both basic and acidic functional groups and can therefore be converted into either a base addition salt or an acid addition salt.

[0032] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing an acid or base using conventional chemical methods. Generally, such salts are prepared by reacting these compounds in the form of free acid or base with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof.

[0033] The term "amino acid" refers to naturally occurring amino acids, synthetic amino acids, and amino acid analogs and mimics that function similarly to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as later modified versions of those amino acids, such as hydroxyproline, γ-carboxyglutamine, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure (e.g., hydrogen, carboxyl group, amino group, and α-carbon bonded to the R group) as naturally occurring amino acids, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs may have a modified R group (e.g., norleucine) or a modified peptide skeleton, but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimics refer to chemical compounds that have a different structure from the general chemical structure of amino acids, but function similarly to naturally occurring amino acids.

[0034] A or Ala as described herein has a structure TIFF0007886963000028.tif15170 represents alanine, and R or Arg indicates that the structure is TIFF0007886963000029.tif18170 represents arginine, and N or Asn indicates that the structure is TIFF0007886963000030.tif16170 represents asparagine, and D or Asp indicates that the structure is TIFF0007886963000031.tif16170 represents aspartic acid, and C or Cys is the structure TIFF0007886963000032.tif16170 represents cysteine, and Q or Gln is structured TIFF0007886963000033.tif16170 represents glutamine, and E or Glu is the structure TIFF0007886963000034.tif16170 represents glutamic acid, and G or Gly indicates that the structure is TIFF0007886963000035.tif11170 represents glycine, and H or His is the structure TIFF0007886963000036.tif18170 represents histidine, and I or Ile has a structure TIFF0007886963000037.tif19170 represents isoleucine, and L or Leu indicates that the structure TIFF0007886963000038.tif16170 represents leucine, and K or Lys indicates that the structure is TIFF0007886963000039.tif17170 represents lysine, and M or Met indicates that the structure is TIFF0007886963000040.tif17170 represents methionine, and F or Phe indicates that the structure is TIFF0007886963000041.tif17170 represents phenylalanine, where P or Pro indicates the structure TIFF0007886963000042.tif17170 represents proline, and S or Ser indicates that the structure TIFF0007886963000043.tif17170 represents serine, and T or Thr indicates that the structure is TIFF0007886963000044.tif17170 represents threonine, and W or Trp is structured TIFF0007886963000045.tif21170 represents tryptophan, and Y or Tyr is structured TIFF0007886963000046.tif19170 represents tyrosine, and V or Val indicates that the structure is This represents valine, which is TIFF0007886963000047.tif17170.

[0035] The term "treatment" includes suppressing, delaying, stopping, or reversing the progression or severity of existing symptoms or disease.

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

[0037] The compounds of the present invention may exist in the form of specific geometric isomers or stereoisomers. The present invention envisions all such compounds, encompassing cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic and other mixtures thereof, such as mixtures rich in enantiomers or diastereomers, all of which are within the scope of the present invention. Substituents such as alkyl groups may have additional chiral carbon atoms. All of these isomers and mixtures thereof are within the scope of the present invention.

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

[0039] Unless otherwise specified, the terms "cis-trans isomer" or "geometric isomer" refer to isomers resulting from the inability of a double bond or a single bond between ring-forming carbon atoms to rotate freely.

[0040] Unless otherwise specified, the term "diastereomer" refers to stereoisomers in which a molecule has two or more chiral centers and is in a non-mirror image relationship with the other molecules.

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

[0042] Unless otherwise specified, wedge-shaped solid line connections ( TIFF0007886963000048.tif4170) and wedge-shaped dashed line connection ( TIFF0007886963000049.tif4170) represents the absolute configuration of a single solid center, and is connected by a linear solid line ( TIFF0007886963000050.tif4170) and linear dashed line combination ( TIFF0007886963000051.tif5170) represents the relative arrangement of the center of the 3D object, and the wavy line ( TIFF0007886963000052.tif4170) uses a wedge-shaped solid line connection ( TIFF0007886963000053.tif4170) or wedge-shaped dashed line connection ( TIFF0007886963000054.tif4170) represents, or a wavy line ( Linear solid line connection (TIFF0007886963000055.tif4170) TIFF0007886963000056.tif4170) or linear dashed line combination ( This represents TIFF0007886963000057.tif5170).

[0043] Unless otherwise specified, the terms “rich in one isomer,” “rich in isomers,” “rich in one enantiomer,” or “rich in enantiomers” mean that the content of one isomer or enantiomer therein is less than 100%, but 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 "isomer excess" or "enantiomer excess" refers to the difference in relative percentages between two isomers or enantiomers. For example, if one isomer or enantiomer is present in a 90% solution and the other isomer or enantiomer is present in a 10% solution, the isomer or enantiomer 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. To obtain one enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or by induction with a chiral auxiliary agent, where the resulting diastereomer mixture is separated and the auxiliary groups are decomposed to provide the required pure enantiomer. Alternatively, if the molecule contains a basic functional group (e.g., an amino group) or an acidic functional group (e.g., a carboxyl group), a salt of the diastereomer is formed with a suitable optically active acid or base, and then the diastereomer is separated by conventional methods known in the art and recovered to obtain the pure enantiomer. Furthermore, the separation of enantiomers and diastereomers is usually performed by chromatography, using a chiral stationary phase and optionally in combination with a chemical induction method (e.g., generating a carbamate from an amine).

[0046] The compounds of the present invention may contain one or more atoms constituting the compound that are atomic isotopes in unnatural proportions. For example, tritium ( 3 H), Iodine-125( 125 I) or C-14 ( 14 Compounds can be labeled with radioactive isotopes such as C). Furthermore, for example, hydrogen can be replaced with deuterium to form deuterated pharmaceuticals. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared to undeuterated pharmaceuticals, deuterated pharmaceuticals offer advantages such as reduced toxicity and side effects, increased drug stability, enhanced therapeutic effect, and extended biological half-life. All isotopic transformations of the compounds of this invention, whether radioactive or not, are included within the scope of this invention.

[0047] If the linking direction is not indicated for the listed linking groups, the linking direction is arbitrary, for example. If the linking group L in TIFF0007886963000058.tif13170 is -MW-, then -MW- links ring A and ring B in the same direction as the reading order from left to right. TIFF0007886963000059.tif15170 can be constructed, and ring A and ring B can be concatenated in the opposite direction to the left-to-right reading order. TIFF0007886963000060.tif15170 can be constructed. The above combinations of linking groups, substituents and / or their variants are permitted only if such combinations produce a stable compound.

[0048] Unless otherwise specified, if a group has one or more connectable sites, one or more of these sites can be connected to other groups via chemical bonds. If the mode of connection of these chemical bonds is omnidirectional and there are H atoms at the connectable sites, when the chemical bonds are connected, the number of H atoms at the sites decreases in proportion to the number of chemical bonds connected, forming a group with the corresponding valency. The chemical bonds between the above sites and other groups are linear solid bonds. TIFF0007886963000061.tif5170), linear dashed line connection ( TIFF0007886963000062.tif5170), or wavy line ( It can be represented as TIFF0007886963000063.tif5170). For example, the linear solid bond of -OCH3 represents a link to another group via the oxygen atom of that group. The linear dashed bond in TIFF0007886963000064.tif7170 indicates that the nitrogen atom of that group is linked to other groups at both ends. The wavy lines in TIFF0007886963000065.tif13170 indicate that the phenyl group is linked to other groups via the carbon atoms at positions 1 and 2.

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

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

[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, the embodiments combined with other chemical synthesis methods, and equivalent alternative methods well-known to those skilled in the art. 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 this invention, the following abbreviations are used: aq represents water, eq represents equivalent, DCM represents dichloromethane, PE represents petroleum ether, DMSO represents dimethyl sulfoxide, MeOH represents methanol, BOC represents the amine protecting group tert-butoxycarbonyl, rt represents room temperature, O / N represents overnight, THF represents tetrahydrofuran, and Boc2O represents di-te-dicarbonate. rt-butyl represents, TFA represents trifluoroacetic acid, DIEA represents diisopropylethylamine, DMF represents N,N-dimethylformamide, HBTU represents benzotriazole-N,N,N',N'-tetramethyluronium-hexafluorophosphate, HOBT represents 1-hydroxybenzotriazole, HOAT represents 1-hydroxy-7-azabenzotriazole, DIC represents N,N'-diisopropylcarbodiimide, DBU represents 1,8-diazabicyclo[5.4.0]undeca-7-ene, PhSiH3 represents phenylsilane, and Pd(PPh3)4 represents tetrakis(triphenylphosphine)palladium. [Modes for carrying out the invention]

[0054] The present invention will be described in detail below with reference to examples, but this is not intended to imply any disadvantageous limitations on the present invention. While the present invention is described in detail in this specification, specific embodiments are also disclosed, and it will be apparent to those skilled in the art that various modifications and improvements can be made to specific embodiments of the present invention without departing from the spirit and scope of the invention. Intermediate A-1 TIFF0007886963000066.tif119170 Step 1: Resin Immobilization 1.1 Weigh 4 g of chloro(o-chlorophenyl)diphenylmethane (2-CTC Resin (degree of substitution S=1.00 mmol / g)) and 1.54 g of A-1_1 into the reaction column, add 25 mL of DCM, then add 3 mL of N,N-diisopropylethylamine to the reaction column and pass nitrogen gas through for 2 hours, then add 4 mL of MeOH to the reaction column and continue to pass nitrogen gas through for 30 minutes, draining the waste liquid until no more liquid comes out, add 50 mL of DMF and wash 5 times for 1 minute each time, draining the waste liquid until no more liquid comes out. 1.2 A 20% piperidine / DMF (50 mL) solution was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. DMF (50 mL) was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. Step 2: Amino acid coupling 2.1 Coupling of A-1_1 a) A-1_1 (3.0 eq) was weighed and added to the above resin, and DIEA (6.00 eq) and 20 mL of DMF were added to the reaction column. Nitrogen gas was passed through, and after the amino acids 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, and ninhydrin was used for detection. The resin was colorless and transparent. c) The reaction solution was extracted and washed five times with DMF for one minute each time, and the waste liquid was drained until no more liquid came out. 2.2 Coupling of A-1_a a) 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. b) A-1_a (3.0 eq) was weighed and added to the above resin. The reaction column was then supplemented with DIEA (3.00 eq) and 20 mL of DMF. Nitrogen gas was passed through the column, and after the amino acids had dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure that the resin expanded uniformly. 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 five times with DMF (50 mL) for 1 minute each time, and waste liquid was drained until no more liquid came out. 2.3 Coupling of A-1_b a) 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was passed through for 20 minutes, and waste liquid was drained until no more liquid came out. DMF (50 mL) was added and washed five times for 1 minute each time, and waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. b) A-1_b (3.0 eq) was weighed and added to the above resin. The reaction column was then supplemented with DIEA (6.00 eq) and 20 mL of DMF. Nitrogen gas was passed through the column, and after the amino acids had dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure that the resin expanded uniformly. 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 mixture was extracted and washed five times with DMF for one minute each time, and waste liquid was discharged until no more liquid came out. Step 3: Cutting and drying of crude peptides 3.1 The cutting fluid was prepared in the following quantities. TIFF0007886963000067.tif251703.2 60 mL of the prepared cutting solution was poured into a reactor containing dried peptide resin, the reactor was aerated for 20 minutes, filtered, and the filtrate was added to a flask. This procedure was repeated twice, and the cutting solutions collected in the two procedures were spin-dried to obtain A-1. Intermediate A-2 Referencing the synthesis of intermediate A-1 (TIFF0007886963000068.tif25170), intermediate A-2 was obtained. Step 1. Immobilization of the resin 1.1 Weigh 4 g of chloro-(o-chlorophenyl)diphenylmethane resin (degree of substitution S = 1.00 mmol / g) and 1.54 g of Fmoc-AEEA-OH into the reaction column. Add 25 mL of DCM, then 3 mL of DIEA and pass nitrogen gas through for 2 hours. Next, add 4 mL of MeOH and continue to pass nitrogen gas through for 30 minutes. Discharge the waste liquid until no more liquid comes out. Add 50 mL of DMF and wash 5 times for 1 minute each time, and discharge the waste liquid until no more liquid comes out. 1.2 A 20% piperidine / DMF (50 mL) solution was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. DMF (50 mL) was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. Step 2. Amino acid coupling 2.1 Coupling of Fmoc-AEEA-OH 1. Weigh Fmoc-AEEA-OH (3.00 eq) and add it to the above resin. Add DIEA (6.00 eq) and 20 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand uniformly. 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 five times with DMF for one minute each time, and waste liquid was drained until no more liquid came out. 2.2 Fmoc-Glu-OtBu Coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Glu-OtBu (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (3.00 eq) and 20 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 waste liquid was drained until no more liquid came out. 2.3 Coupling of 20-(tBu)-eicosanedionic acid 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was passed through for 20 minutes, and waste liquid was drained until no more liquid came out. DMF (50 mL) was added and washed 5 times for 1 minute each time, and waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed 20-(tBu)-eicosanedionic acid (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 20 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 for one minute each time, and waste liquid was drained until no more liquid came out. Step 3: Cutting and drying of crude peptides 3.1 The cutting fluid was prepared in the following quantities. TIFF0007886963000069.tif251703.2 This operation was repeated twice, and the two collected cutting fluids were spin-dried to obtain A-2. Intermediate A-3 Referring to the synthesis of intermediate A-1 (TIFF0007886963000070.tif27170), intermediate A-3 was obtained. Intermediate A-4 Referring to the synthesis of intermediate A-1 (TIFF0007886963000071.tif27170), intermediate A-4 was obtained.

[0055] Example 1 TIFF0007886963000072.tif52170

[0056] Step 1: Weigh 1.34 g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (substitution degree Sub=0.3 mmol / g), place it in the reaction column, add 50 mL of DMF, and circulate nitrogen gas for 2 hours. Discharge waste liquid until no more liquid comes out, add 50 mL of DMF and wash 5 times for 1 minute each time, and discharge waste liquid until no more liquid comes out.

[0057] Step 2: 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was passed through for 20 minutes, and waste liquid was drained until no more liquid came out. DMF (50 mL) was added and washed five times for 1 minute each time, and 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 Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin expands 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 five times with DMF (50 mL each time) for 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF for one minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0063] 3.5 Fmoc-Ala-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ala-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gly-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ser(tBu)-OH (3.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ser(tBu)-OH (3.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0068] 3.10 Fmoc-Gly-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gly-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0069] 3.11 Fmoc-Gly-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gly-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0070] 3.12 Fmoc-Ala-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ala-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0071] 3.13 Fmoc-Ile-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ile-OH (3.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0072] 3.14 Fmoc-Leu-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Leu-OH (3.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0073] 3.15 Fmoc-Trp(Boc)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Trp(Boc)-OH (3.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0074] 3.16 Fmoc-Lys(Dde)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Lys(Dde)-OH (3.0 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0075] 3.17 Fmoc-Val-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Val-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0076] 3.18 Fmoc-Phe-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Phe-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0077] 3.19 Fmoc-Ala-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ala-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0078] 3.20 Fmoc-Lys(Alloc)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Lys(Alloc)-OH (2.0 eq) and add it to the above resin. Add HOBT (2.00 eq) and 10 mL of DMF to the reaction column, and pass nitrogen gas through. After the amino acids and HOBT have dissolved, add DIC (2.00 eq). The nitrogen gas was adjusted so that the resin expands uniformly. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Gln(Trt)-OH (3.0 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin expands uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0080] 3.22 Fmoc-Ala-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ala-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0081] 3.23 Fmoc-Ile-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ile-OH (3.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0082] 3.24 Fmoc-Lys(Boc)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Lys(Boc)-OH (2.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0083] 3.25 Fmoc-Asp(OtBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Asp(OtBu)-OH (6.0 eq) and add it to the above resin. Add DIEA (12.0 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin expands uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0084] 3.26 Fmoc-Leu-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Leu-OH (6.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Aib-OH (6.0 eq) and add it to the resin mentioned above. Add DIEA (12.0 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0086] 3.28 Fmoc-Ile-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Ile-OH (6.0 eq) and add it to the above resin. Add HOAT (6.00 eq) and 10 mL of DMF to the reaction column, and pass nitrogen gas through. After the amino acids and HOAT have dissolved, add DIC (6.00 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ser(tBu)-OH (6.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0088] 3.30 Fmoc-Tyr(tBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Tyr(tBu)-OH (6.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0089] 3.31 Fmoc-Asp(OtBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Asp(OtBu)-OH (6.0 eq) and add it to the above resin. Add DIEA (12.0 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HATU (5.70 eq). The nitrogen gas was adjusted so that the resin expands uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0090] 3.32 Fmoc-Ser(tBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ser(tBu)-OH (6.0 eq) and added it to the above resin. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Thr(tBu)-OH (6.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0092] 3.34 Fmoc-Phe-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Phe-OH (6.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0093] 3.35 Fmoc-Thr(tBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Thr(tBu)-OH (6.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (12.0 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0094] 3.36 Fmoc-Gly-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gly-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Glu(OtBu)-OH (6.0eq) and added it to the above resin. Replenished the reaction column with HOBT (6.00eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids and HOBT dissolved, DIC (6.00eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0096] 3.38 Fmoc-Aib-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Aib-OH (3.0 eq) and add it to the resin mentioned above. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0097] 3.39 Boc-Tyr(tBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Boc-Tyr(tBu)-OH (6.0 eq) was weighed and added to the above resin. DIEA (12.0 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids dissolved, HATU (5.70 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and 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 the reaction column, nitrogen gas was passed through, then Pd(PPh3)4 (0.1 eq) was added, nitrogen gas was passed through for 20 minutes, and the reaction was repeated twice. Waste liquid was drained until no more liquid came out. 2. Wash with DMF five times (50 mL each time) for 1 minute each time, and drain the waste liquid until no more liquid comes out.

[0099] 3.41 Fmoc-Ida-OH coupling 1. Weigh Fmoc-Ida-OH (4.0 eq) and add it to the resin. Add DIEA (8.00 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HBTU (3.80 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 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. 50 mL of 10% DBU / DMF was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Intermediate A-1 (1.50 eq) was weighed and added to the above resin. DIEA (3.00 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids dissolved, HBTU (1.45 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

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

[0102] 3.44 Closure of the amide ring 1. DIEA (3.0 eq) was added to the resin in the DMF solution, and then HATU (1.5 eq) dissolved in DMF was slowly added dropwise to the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin expanded 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 five times with DMF (50 mL each time) for 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 minutes each time, and the waste liquid was drained until no more liquid came out. The resin was then removed, dried, and stored as a reserve.

[0103] Step 4. Cutting and drying of crude peptides

[0104] 4.1 The cutting fluid was prepared in the following quantities. TIFF0007886963000073.tif30170

[0105] 4.2 The dried peptide resin was placed in the prepared cutting solution, shaken in a shaker for 2.5 hours, filtered, and the filtrate was added to 10 times the volume of ice-cold isopropyl ether, centrifuged, and washed 5 times with isopropyl ether. The crude peptide was vacuum-dried for 2 hours and purified to obtain polypeptide compound WX-001. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value [M+4H] / 4 of 1228.6 and a detected value of 1228.7.

[0106] Example 2 Referencing the synthesis of TIFF0007886963000074.tif52170WX-001, polypeptide WX-002 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1243.1 and a detected value of 1242.8.

[0107] Example 3 Referencing the synthesis of TIFF0007886963000075.tif54170WX-001, polypeptide WX-003 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1232.1 and a detected value of 1232.2.

[0108] Example 4 Referencing the synthesis of TIFF0007886963000076.tif54170WX-001, polypeptide WX-004 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1228.6 and a detected value of 1228.2.

[0109] Example 5 TIFF0007886963000077.tif55170

[0110] Step 1: Weigh 1.34 g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (substitution degree Sub=0.3 mmol / g), place it in the reaction column, add 50 mL of DMF, and circulate nitrogen gas for 2 hours. Discharge waste liquid until no more liquid comes out, add 50 mL of DMF, wash 5 times for 1 minute each time, and discharge waste liquid until no more liquid comes out.

[0111] Step 2: 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was passed through for 20 minutes, and waste liquid was drained until no more liquid came out. DMF (50 mL) was added and washed five times for 1 minute each time, and 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 Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin expands 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 five times with DMF (50 mL each time) for 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out. 5. By repeating step 3.2, the following amino acid coupling was completed. TIFF0007886963000078.tif234170

[0115] 3.40 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (10 mL) were placed in the reaction column, nitrogen gas was passed through, then Pd(PPh3)4 (0.1 eq) was added, nitrogen gas was passed through for 20 minutes, and the reaction was repeated twice. Waste liquid was drained until no more liquid came out. 2. Wash with DMF five times (50 mL each time) for 1 minute each time, and drain the waste liquid until no more liquid comes out.

[0116] 3.41 Fmoc-Ida-OH coupling 1. Weigh Fmoc-Ida-OH (4.0 eq) and add it to the resin. Add DIEA (8.00 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HBTU (3.80 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 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. 50 mL of 10% DBU / DMF was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Intermediate A-1 (1.50 eq) was weighed and added to the above resin. DIEA (3.00 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids dissolved, HBTU (1.45 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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 passed through for 15 minutes, waste liquid was drained, and DMF (50 mL) was added and washed five times for 1 minute each time, waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0119] 3.44 Closure of the amide ring 1. DIEA (3.0 eq) was added to the resin in the DMF solution, and then HATU (1.5 eq) dissolved in DMF was slowly added dropwise to the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin expanded 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 five times with DMF (50 mL each time) for 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 minutes each time, and the waste liquid was drained until no more liquid came out. The resin was then removed, dried, and stored as a reserve.

[0120] Step 4. Cutting and drying of crude peptides

[0121] 4.1 The cutting fluid was prepared in the following quantities. TIFF0007886963000079.tif30170

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

[0123] Example 6 Referencing the synthesis of TIFF0007886963000080.tif49170WX-001, polypeptide WX-006 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1232.4 and a detected value of 1232.4.

[0124] Example 7 Referencing the synthesis of TIFF0007886963000081.tif48170WX-001, polypeptide WX-007 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1218.1 and a detected value of 1218.3.

[0125] Example 8 Referencing the synthesis of TIFF0007886963000082.tif49170WX-001, polypeptide WX-008 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1228.6 and a detected value of 1228.6.

[0126] Example 9 TIFF0007886963000083.tif50170

[0127] Step 1: Weigh 1.34 g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (substitution degree Sub=0.3 mmol / g), place it in the reaction column, add 50 mL of DMF, and circulate nitrogen gas for 2 hours. Discharge waste liquid until no more liquid comes out, add 50 mL of DMF, wash 5 times for 1 minute each time, and discharge waste liquid until no more liquid comes out.

[0128] Step 2: 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was passed through for 20 minutes, and waste liquid was drained until no more liquid came out. DMF (50 mL) was added and washed five times for 1 minute each time, and 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 Fmoc-Ser(tBu)-OH (3.0 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin expands 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 five times with DMF (50 mL each time) for 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.0 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out. 5. By repeating step 3.2, the following amino acid coupling was completed. TIFF0007886963000084.tif234170

[0132] 3.40 De-Alloc 1. PhSiH3 (10.0 eq) and DCM (10 mL) were placed in the reaction column, nitrogen gas was passed through, then Pd(PPh3)4 (0.1 eq) was added, nitrogen gas was passed through for 20 minutes, and the reaction was repeated twice. Waste liquid was drained until no more liquid came out. 2. Wash with DMF five times (50 mL each time) for 1 minute each time, and drain the waste liquid until no more liquid comes out.

[0133] 3.41 Fmoc-Ida-OH coupling 1. Weigh Fmoc-Ida-OH (4.0 eq) and add it to the above resin. Add DIEA (8.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (3.80 eq). The nitrogen gas was adjusted so that the resin expands 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 five times with DMF (50 mL each time) for 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. 50 mL of 10% DBU / DMF was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Intermediate A-1 (1.50 eq) was weighed and added to the above resin. DIEA (3.00 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids dissolved, HBTU (1.45 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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 passed through for 15 minutes, waste liquid was drained, and DMF (50 mL) was added and washed five times for 1 minute each time, waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue.

[0136] 3.44 Closure of the amide ring 1. DIEA (3.0 eq) was added to the resin in the DMF solution, and then HATU (1.5 eq) dissolved in DMF was slowly added dropwise to the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin expanded 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 five times with DMF (50 mL each time) for 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 minutes each time, and the waste liquid was drained until no more liquid came out. The resin was then removed, dried, and stored as a reserve.

[0137] Step 4. Cutting and drying of crude peptides

[0138] 4.1 The cutting fluid was prepared in the following quantities. TIFF0007886963000085.tif30170

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

[0140] Example 10 TIFF0007886963000086.tif57170

[0141] Step 1: Weigh 1.08 g of 4-(2',4'-dimethoxyphenyl-fluorenemethoxycarbonyl-aminomethyl)-phenoxyacetamide-methylbenzhydrylamine resin (substitution degree Sub=0.37 mmol / g), place it in the reaction column, add 50 mL of DMF, and flush with nitrogen gas for 2 hours. Discharge waste liquid until no more liquid comes out, add 50 mL of DMF and wash 5 times for 1 minute each time, and discharge waste liquid until no more liquid comes out.

[0142] Step 2: 20% piperidine / DMF (50 mL) was placed in the reaction column, nitrogen gas was passed through for 20 minutes, and waste liquid was drained until no more liquid came out. DMF (50 mL) was added and washed five times for 1 minute each time, and 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 Fmoc-Ser(tBu)-OH (3.00 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin expands 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 five times with DMF (50 mL each time) for 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Pro-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0148] 3.5 Fmoc-Ala-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ala-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gly-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ser(tBu)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0151] 3.8 Coupling of Fmoc-Ser(tBu)-OH 1. Put 20% piperidine / DMF (50 mL) into the reaction column, ventilate with nitrogen gas for 20 minutes, and 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. Ninhydrin was used for detection, and the resin looked blue. 2. Weigh Fmoc-Ser(tBu)-OH (3.00 eq) and add it to the above resin. Supplement the reaction column with DIEA (6.00 eq) and 10 mL DMF, ventilate with nitrogen gas, and add HBTU (2.85 eq) after the amino acid is dissolved. The nitrogen gas was adjusted so that the resin swelled uniformly. 3. The reaction was carried out for 0.5 hours at 25 °C. Ninhydrin was used for detection, and the resin was colorless and transparent. 4. Withdraw the reaction solution and wash it 5 times (50 mL each time) with DMF, 1 minute each time, and drain the waste liquid until no liquid comes out.

[0152] 3.9 Coupling of Fmoc-Pro-OH 1. Put 20% piperidine / DMF (50 mL) into the reaction column, ventilate with nitrogen gas for 20 minutes, and 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. Ninhydrin was used for detection, and the resin looked blue. 2. Weigh Fmoc-Pro-OH (3.00 eq) and add it to the above resin. Supplement the reaction column with DIEA (6.00 eq) and 10 mL DMF, ventilate with nitrogen gas, and add HBTU (2.85 eq) after the amino acid is dissolved. The nitrogen gas was adjusted so that the resin swelled uniformly. 3. The reaction was carried out for 0.5 hours at 25 °C. Ninhydrin was used for detection, and the resin was colorless and transparent. 4. Withdraw the reaction solution and wash it 5 times (50 mL each time) with DMF, 1 minute each time, and drain the waste liquid until no liquid comes out.

[0153] 3.10 Coupling of Fmoc-Gly-Gly-OH 1. Put 20% piperidine / DMF (50 mL) into the reaction column, purge with nitrogen gas for 20 minutes, and 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. Chloranil was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Gly-Gly-OH (3.00 eq) and add it to the above resin. Supplement the reaction column with DIEA (6.00 eq) and 10 mL DMF, purge with nitrogen gas, and add HBTU (2.85 eq) after the amino acid is dissolved. The nitrogen gas was adjusted so that the resin swelled uniformly. 3. The reaction was carried out for 0.5 hours at 25 °C. Chloranil was used for detection, and the resin was colorless and transparent. 4. Withdraw the reaction solution, wash 5 times (50 mL each time) with DMF, 1 minute each time, and drain the waste liquid until no liquid comes out.

[0154] 3.11 Coupling of Fmoc-Ala-OH 1. Put 20% piperidine / DMF (50 mL) into the reaction column, purge with nitrogen gas for 20 minutes, and 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. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Ala-OH (3.00 eq) and add it to the above resin. Supplement the reaction column with DIEA (6.00 eq) and 10 mL DMF, purge with nitrogen gas, and add HBTU (2.85 eq) after the amino acid is dissolved. The nitrogen gas was adjusted so that the resin swelled uniformly. 3. The reaction was carried out for 0.5 hours at 25 °C. Ninhydrin was used for detection, and the resin was colorless and transparent. 4. Withdraw the reaction solution, wash 5 times (50 mL each time) with DMF, 1 minute each time, and drain the waste liquid until no liquid comes out.

[0155] 3.12 Coupling of Fmoc-Ile-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ile-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0156] 3.13 Fmoc-Leu-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Leu-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0157] 3.14 Fmoc-Trp(Boc)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Trp(Boc)-OH (3.00 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin expands uniformly. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Glu(OtBu)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0159] 3.16 Fmoc-Val-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Val-OH (3.00 eq) and added it to the above resin. Then, DIEA (6.00 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids had dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin would expand uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0160] 3.17 Fmoc-Phe-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Phe-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0161] 3.18 Fmoc-Ala-OH coupling 1. Put 20% piperidine / DMF (50 mL) into the reaction column, purge with nitrogen gas for 20 minutes, and drain the waste liquid until no liquid comes out. Add DMF (50 mL) and wash 5 times, each time for 1 minute, and drain the waste liquid until no liquid comes out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Ala-OH (3.00 eq) and add it to the above resin. Supplement the reaction column with DIEA (6.00 eq) and 10 mL DMF, purge with nitrogen gas, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled uniformly. 3. The reaction was carried out for 0.5 hours under an environment of 25 °C. Ninhydrin was used for detection, and the resin was colorless and transparent. 4. Withdraw the reaction solution, wash it 5 times (50 mL each time) with DMF, each time for 1 minute, and drain the waste liquid until no liquid comes out.

[0162] 3.19 Coupling of Fmoc-Lys(Alloc)-OH 1. Put 20% piperidine / DMF (50 mL) into the reaction column, purge with nitrogen gas for 20 minutes, and drain the waste liquid until no liquid comes out. Add DMF (50 mL) and wash 5 times, each time for 1 minute, and drain the waste liquid until no liquid comes out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Lys(Alloc)-OH (3.00 eq) and add it to the above resin. Supplement the reaction column with DIEA (6.00 eq) and 10 mL DMF, purge with nitrogen gas, and after the amino acid is dissolved, add HBTU (2.85 eq). The nitrogen gas was adjusted so that the resin swelled uniformly. 3. The reaction was carried out for 0.5 hours under an environment of 25 °C. Ninhydrin was used for detection, and the resin was colorless and transparent. 4. Withdraw the reaction solution, wash it 5 times (50 mL each time) with DMF, each time for 1 minute, and drain the waste liquid until no liquid comes out.

[0163] 3.20 Coupling of Fmoc-Gln(Trt)-OH 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gln(Trt)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0164] 3.21 Fmoc-Ala-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ala-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HBTU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0165] 3.22 Fmoc-Lys(Dde)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Lys(Dde)-OH (3.00 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin expands uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0166] 3.23 Fmoc-Lys(Boc)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Lys(Boc)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0167] 3.24 Fmoc-Asp(OtBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Asp(OtBu)-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0168] 3.25 Fmoc-Leu-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Leu-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Aib-OH (3.00 eq) and add it to the resin mentioned above. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0170] 3.27 Fmoc-Ile-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ile-OH (3.00 eq) and added it to the above resin. Then, DIEA (6.00 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 five times with DMF (50 mL each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Ser(tBu)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted so that the resin expanded uniformly. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Tyr(tBu)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0173] 3.30 Fmoc-Asp(OtBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Asp(OtBu)-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Thr(tBu)-SerPsi(Me,Me)Pro-OH (2.00 eq) and add it to the above resin. Add DIEA (4.00 eq) and 10 mL DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HATU (1.90 eq). The nitrogen gas was adjusted so that the resin would expand uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0175] 3.32 Fmoc-Phe-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Phe-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and passed nitrogen gas through. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Thr(tBu)-OH (3.00 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin would expand uniformly. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0177] 3.34 Fmoc-Gly-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Gly-OH (3.00 eq) and added it to the resin mentioned above. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and a nitrogen gas was passed through. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weighed Fmoc-Glu(OtBu)-OH (3.00 eq) and added it to the above resin. Replenished the reaction column with DIEA (6.00 eq) and 10 mL of DMF, and aeration with nitrogen gas. After the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0179] 3.36 Fmoc-Aib-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh Fmoc-Aib-OH (3.00 eq) and add it to the resin mentioned above. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 five times with DMF (50 mL each time) for 1 minute each time, and waste liquid was drained until no more liquid came out.

[0180] 3.37 Boc-Tyr(tBu)-OH coupling 1. 20% piperidine / DMF (50 mL) was placed in the reaction column, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Chloranil was used for detection, and the resin appeared blue. 2. Boc-Tyr(tBu)-OH (3.00 eq) was weighed and added to the above resin. DIEA (6.00 eq) and 10 mL of DMF were added to the reaction column, nitrogen gas was passed through, and after the amino acids dissolved, HATU (2.85 eq) was added. The nitrogen gas was adjusted to ensure uniform expansion of the resin. 3. The reaction was carried out at 25°C for 0.5 hours, and chloranil was used for detection. The resin was colorless and transparent. 4. The reaction solution was removed and washed five times with DMF (50 mL each time) for 1 minute each time, and 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 the reaction column, nitrogen gas was passed through, then Pd(PPh3)4 (0.10 eq) was added, nitrogen gas was passed through for 20 minutes, and the reaction was repeated twice. Waste liquid was drained until no more liquid came out. 2. Wash with DMF five times (50 mL each time) for 1 minute each time, and drain the waste liquid until no more liquid comes out.

[0182] 3.39 Fmoc-Glu-OAll Coupling 1. Weigh Fmoc-Glu-OAll (3.00 eq) and add it to the above resin. Add DIEA (6.00 eq) and 10 mL of DMF to the reaction column, pass nitrogen gas through, and after the amino acids have dissolved, add HATU (2.85 eq). The nitrogen gas was adjusted so that the resin expands 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 five times with DMF (50 mL each time) for 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, and nitrogen gas was passed through for 20 minutes. Waste liquid was drained until no more liquid came out. 50 mL of DMF was added and the column was washed 5 times for 1 minute each time. Waste liquid was drained until no more liquid came out. Ninhydrin was used for detection, and the resin appeared blue. 2. Weigh intermediate A-1 (1.50 eq) and add it to the resin mentioned above. Add DIEA (3.00 eq) and 10 mL of DMF to the reaction column, then pass nitrogen gas through. After the amino acids have dissolved, add HATU (1.45 eq). The nitrogen gas was adjusted to ensure uniform expansion of the resin. 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 each time) for 1 minute each time, and 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 the reaction column, nitrogen gas was passed through, then Pd(PPh3)4 (0.10 eq) was added, nitrogen gas was passed through for 20 minutes, and the reaction was repeated twice. Waste liquid was drained until no more liquid came out. 2. Wash with DMF five times (50 mL each time) for 1 minute each time, and drain the waste liquid until no more liquid comes out.

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

[0186] 3.43 Closure of the amide ring 1. DIEA (3.00 eq) was added to the resin in the DMF solution, and then HATU (1.50 eq) dissolved in DMF was slowly added dropwise to the reaction column, and nitrogen gas was passed through. The nitrogen gas was adjusted so that the resin expanded 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 five times with DMF (50 mL each time) for 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 minutes each time, and the waste liquid was drained until no more liquid came out. The resin was then removed, dried, and stored as a reserve.

[0187] Step 4. Cutting and drying of crude peptides

[0188] 4.1 The cutting fluid was prepared in the following quantities. TIFF0007886963000087.tif30170

[0189] 4.2 The dried peptide resin was placed in the prepared cutting solution, shaken in a shaker for 2.5 hours, filtered, and the filtrate was added to 10 times the volume of ice-cold isopropyl ether. The filtrate was 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, with a calculated value of 1236.1 and a detected value of 1236.0.

[0190] Example 11 Referencing the synthesis of TIFF0007886963000088.tif62170WX-010, polypeptide WX-011 was obtained. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1236.1 and a detected value of 1236.0.

[0191] Example 12 Referring to the synthesis of TIFF0007886963000089.tif55170WX-001, polypeptide WX-012 was obtained using intermediate A-2. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1225.6 and a detected value of 1225.7.

[0192] Example 13 Referring to the synthesis of TIFF0007886963000090.tif59170WX-001, polypeptide WX-013 was obtained using intermediate A-3. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1264.9 and a detected value of 1264.6.

[0193] Example 14 Referring to the synthesis of TIFF0007886963000091.tif54170WX-001, polypeptide WX-014 was obtained using intermediate A-4. The molecular weight of the polypeptide was confirmed by ESI-MS, with a calculated value of [M+4H] / 4 of 1257.9 and a detected value of 1257.8.

[0194] Biological test data

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

[0196] A: Main ingredients: 1) Cell line This cell line was constructed by Wuxi APPTEC (Shanghai) Co., Ltd. See Table 1 below for details. TIFF0007886963000092.tif241702) Reagents and Consumables TIFF0007886963000093.tif791703) Equipment TIFF0007886963000094.tif44170

[0197] B. Method 1) Experimental materials Experimental buffer TIFF0007886963000095.tif60170 Preparation of assay reagents TIFF0007886963000096.tif331702) Experimental Method a) Preparation of compound plates: The test compound was diluted by Bravo at a 4x multiplier of 10 points, starting from an initial concentration of 30 μM. b) Transfer of compounds: 1) 100 nL of the compound was transferred to an OptiPlate-384 plate using 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 by placing it in warm water at 37°C. 2) The cell suspension was transferred to a 15 mL centrifuge tube and lightly washed with 10 mL of HBSS. 3) The centrifuge 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 lightly washed with 10 mL of HBSS, the cells were centrifuged to settle, and finally the cells were resuspended in the experimental buffer. 6) Cell density and viability were measured using Vi-cell. 7) GLP-1R / GIPR cells at a concentration of 2.0*10 5 Diluted with experimental buffer to a concentration of / mL. 8) 100 nL of 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 peeled off and read by EnVision.

[0198] C. Experimental Results The experimental results are shown in Table 6. TIFF0007886963000097.tif99170 Conclusion: The compounds of the present invention exhibit very strong agonist activity against GLP-1R / GIPR.

[0199] Test Example 2: Evaluation of the pharmacokinetic profile of a compound in rats A. Experimental Objectives The purpose was to test the pharmacokinetic profile of compounds in SD rats. B. Experimental Procedure The pharmacokinetic properties of candidate compounds in rodents after subcutaneous injection were investigated using a standard protocol. In the experiment, candidate compounds were formulated into clear solutions 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, prepared as plasma, and the drug concentrations were analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using Phoenix WinNonlin software. C. Experimental Results The experimental results are shown in Table 7. TIFF0007886963000098.tif33170 Conclusion: The compounds of the present invention have excellent pharmacokinetic properties in rats.

[0200] Test Example 3: Evaluation of the pharmacokinetic profile of a compound in mice A. Experimental Objectives The purpose was to test the pharmacokinetic profile of the compound in C57BL / 6 mice. B. Experimental Procedure The pharmacokinetic properties of candidate compounds in rodents after subcutaneous injection were investigated using a standard protocol. In the experiment, candidate compounds were formulated into clear solutions and administered to rats by subcutaneous injection (SC, 0.048 mpk). The subcutaneous injection vehicle was citrate buffer (20 mM, pH=7). Whole blood was collected, prepared as plasma, and the drug concentrations were analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using Phoenix WinNonlin software. C. Experimental Results The experimental results are shown in Table 8. TIFF0007886963000099.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. Experimental Objectives The purpose was to test the pharmacokinetic profile of the compound in cynomolgus monkeys. B. Experimental Procedure The pharmacokinetic properties of the compounds in mammals after intravenous and subcutaneous administration were investigated using a standard protocol. In the experiment, candidate compounds were formulated into clear solutions and administered to cynomolgus monkeys by single subcutaneous injection (SC, 0.02 mpk). The subcutaneous injection vehicle was citrate buffer (20 mM, pH=7). Whole blood was collected, prepared as plasma, and the drug concentrations were analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using Phoenix WinNonlin software. C. Experimental Results The experimental results are shown in Table 9. TIFF0007886963000100.tif33170 Conclusion: The compounds of the present invention have excellent pharmacokinetic properties in a sieve.

[0202] Test Example 5: Plasma Stability Test (PLS) A. Experimental Objectives The purpose was to study the stability of test compounds in the plasma of normal mice. B. Experimental Procedure 1. Before the experiment, the coagulated frozen plasma was thawed in a 37°C water bath. The plasma was centrifuged at 4000 rpm for 5 minutes, and if thrombi were present, they were removed, and the pH was adjusted to 7.4 ± 0.1. 2. Preparation of the test compound solution: A 100 μM solution was prepared by diluting with DMSO. 3. 98 μL of blank control plasma was added to 2 μL of test compound solution (100 μM) to a final concentration of 2 μM of the mixture, and this was cultured under water bath conditions at 37°C. 4. At each time point (0, 10, 30, 60, and 120 minutes), 100 μL of H3PO4 solution and 800 μL of stop solution (200 ng / mL tolbutamide and 200 ng / mL labetalol 100% methanol solution) were added, respectively, and the proteins were precipitated and thoroughly mixed. 5. The samples were centrifuged at a rotation speed of 4000 rpm for 20 minutes, and 100 μL of the supernatant was collected from each well and analyzed by LC-MS / MS. C. Experimental Results The experimental results are shown in Table 10. TIFF0007886963000101.tif18170 Conclusion: The compounds of the present invention have excellent plasma stability.

[0203] Test Example 6: Plasma Protein Binding Capacity Test (PPB) A. Experimental Objectives The purpose was to study the binding ability of the test compound to human / mouse plasma albumin. B. Experimental Procedure 1. Matrix preparation (preparation of blank matrix): On the day of the experiment, plasma was thawed in cold water and centrifuged at 3220 rpm for 5 minutes to remove all thrombi. The pH of the obtained plasma was measured and adjusted to 7.4 ± 0.1 with 1% phosphoric acid or 1N sodium hydroxide as needed. 2. Dilution step of test compound: The test compound was dissolved in dimethyl sulfoxide (DMSO) to prepare stock solutions with concentrations of 10 mM and 2 mM, respectively. 2 μL of the stock solution (2 mM) was diluted with 98 μL of DMSO to prepare a 40 μM standard solution. 10 μL of the stock solution was diluted with 240 μL of DMSO to prepare a 400 μM standard solution of the control compound. The standard solution of the compound (5 μL) was uniformly mixed with the blank matrix (995 μL) in a ratio of 1:200 to prepare the weight matrix. 3. Analysis process 3.1 Equal volumes of 30 μL of weighted matrix (n=2) were transferred to a sample collection plate to prepare test time 0 (T0) samples for residue measurement. The samples were immediately combined with the corresponding blank buffer to a final volume of 60 μL in each well, so that the volume ratio of plasma (weighted matrix) to buffer was 1:1. Next, 60 μL of 4% H3PO4 H2O and 480 μL of stop solution containing an internal standard were added to each T0 sample of the test compound. These were then stored at 2–8°C with the other samples for further processing. 3.2 The remaining plasma samples were pre-incubated in a carbon dioxide incubator at 37±1°C for 30 minutes. Protein-free samples (F samples) were prepared and transferred to polycarbonate tubes (n=2) along with the matrix-loaded samples (230 μL), and hypercentrifuged at 37°C and 155,000 × g (35,000 rpm) for 4 hours. 3.3 To prepare the T sample (test sample), one additional matrix-containing sample was transferred to a separate 96-well plate (sample incubation plate) and incubated at 37°C for 4 hours. 3.4 After centrifugation was complete, 30 μL of protein-free sample (F sample) and 30 μL of T sample were collected from the second layer (below the upper layer) of the supernatant 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. Matrix (blank matrix): The volume ratio of buffer was 1:1. 60 μL of 4% H3PO4 aqueous solution and 480 μL of stop solution (containing internal standards) were added to all samples. The mixtures were centrifuged at 4000 rpm for 20 minutes, and 100 μL of supernatant was collected from each sample for LC-MS / MS analysis. C. Experimental Results The experimental results are shown in Table 11. TIFF0007886963000102.tif23170 Note: NA indicates that the plasma protein binding capacity was too high, and the free drug could not be detected at normal plasma protein concentrations. Conclusion: The compounds of the present invention exhibit extremely high plasma protein binding ability.

[0204] Example 7: In vivo efficacy study in an obese mouse model A. Objective of the experiment The weight-reducing effect of the series of polypeptides of the present invention was investigated in obese animal models and compared with commercially available equivalents, tirzepatide and semaglutide. B. Experimental Procedure 1. Modeling of animal models Obese mouse models were obtained by inducing a high-fat diet (MOLECULAR METABOLISM, 18 (2018), P3~14) according to the references. Specifically, the test mice were fed a high-fat diet (Research Diet D12492) for 13 weeks to create the model. While both the normal control group and the model animals gained weight steadily, the model group showed a significantly greater weight increase and significantly higher intake than the control group, suggesting successful modeling. 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 in clear solutions of varying concentrations at room temperature, and then frozen for storage. During the experiment, the animal models and the treatment group were administered the drug by subcutaneous injection into the outer thigh, once every three days (Q3d), for a total of 22 days. Details are as follows. TIFF0007886963000103.tif47170C Results and Analysis During the experiment, all test mice were in good health and mental condition. While the animals in the treatment group showed varying degrees of suppression of food intake due to the effects of the drug, no deaths occurred. After the completion of all treatment cycles, the body weight of the test mice was measured, and the results are shown below. TIFF0007886963000104.tif41170 Conclusion: Both WX001 and WX005 showed good weight loss effects in obese mouse models, with WX005 showing superior efficacy and being better than tilzepatide and semaglutide. In repeated experiments, the test drug showed substantially consistent effects in the corresponding groups, demonstrating the existence of objective efficacy advantages. Furthermore, the beneficial effect of weight loss was also observed with other polypeptide compounds of the present invention.

[0205] Example 8: In vivo efficacy study in a diabetic mouse model A. Objective of the experiment The efficacy of the series of polypeptides of the present invention was investigated in diabetic animal models and compared with the commercially available equivalent, tirzepatide. According to the literature ((MOLECULAR METABOLISM, 18 (2018), P3~14)), tirzepatide's blood glucose-lowering effect is far superior to that of semaglutide; therefore, semaglutide was not included in this experiment for comparison. B. Experimental Procedure 1. Selection of an animal model db / db (Lepr leptin receptor deficiency) mice exhibited typical clinical symptoms of diabetes, such as extreme obesity, polyphagia, thirst, and polyuria, making them an ideal animal model for type 2 diabetes. Specific experimental procedure: db / db test mice weighing over 30g and with fasting blood glucose levels exceeding 15 mmol / L were screened and included in the formal trial. 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 into a clear solution at room temperature, and then frozen for storage. During the experiment, the drug was administered by subcutaneous injection into the outer thigh of the normal control group, model group, and treatment group of animals once daily (Qd) for a total of four weeks. Details are as follows. TIFF0007886963000105.tif41170C Results and Analysis During the experiment, all test mice were in good health and mental condition. While the animals in the treatment group showed varying degrees of suppression of food intake due to the effects of the drug, no deaths occurred. During the experiment, the rate of change in fasting blood glucose and glycated hemoglobin levels in the test mice was dynamically monitored, and the fasting blood glucose levels are shown below. The results for the percentage change in glycated hemoglobin levels were as follows: TIFF0007886963000106.tif47170 TIFF0007886963000107.tif47170 Note: "-" indicates a decrease. Conclusion: Both WX005 and WX009 showed good blood glucose-lowering effects in a mouse model of diabetes. Specifically, both WX005 and WX009 showed similar effects in reducing glycated hemoglobin levels in the test animals, and were superior to tilzepatide. WX005 was slightly superior to WX009 in lowering fasting blood glucose levels in the test animals, and both were superior to tilzepatide. Overall, WX005 and WX009 are considered to have significantly better blood glucose-lowering effects than tilzepatide. In repeated experiments, the test drug showed substantially consistent effects in the corresponding groups, demonstrating the existence of objective efficacy advantages. Furthermore, the advantages in terms of efficacy regarding glucose metabolism in animals were also observed with other polypeptide compounds of the present invention.

Claims

1. The sequence is represented by formulas (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 follows, 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 in formula It is connected to the base indicated by, Here, The structure of Aib is, It is a group represented by, S ０ is, formula Selected from the bases shown, X is the formula Selected from the group consisting of the elements shown, where "*" is X １ It represents the position connected to, X １ These are single bonds, -C(=O)-, -O-C(=O)-, and -N(R １ Selected from the group consisting of )-C(=O), R １ H and C １－３ Selected from alkyl groups, X ２ is, formula Selected from the bases shown, m is selected from 2, 3, and 4. n is selected from 15, 16, 17, 18, and 19. p includes modifications selected from 1 and 2, The use of polypeptides in the preparation of pharmaceuticals for the treatment and / or prevention of diabetes, obesity and related diseases, The aforementioned diabetes-related diseases include diabetic cardiovascular disease, diabetic cerebrovascular disease, retinopathy, nephropathy, cataracts, coronary artery disease, diabetic neuropathy, diabetic foot disease, and peripheral neuropathy, and the aforementioned obesity-related diseases include liver disease, hyperlipidemia, and hypertension, as used in the foregoing.

2. In the polypeptide, X １ The group is selected from the group consisting of single bonds, -C(=O)-, -O-C(=O)-, and -NH-C(=O)-. The use described in claim 1.

3. In the polypeptide described above, n is selected from 15 and 17. The use according to any one of claims 1 to 2.

4. In the polypeptide, X ２ is formula Selected from the group consisting of the groups shown. The use according to any one of claims 1 to 2.

5. In the polypeptide, X ２ is, formula Selected from the group consisting of the groups shown. The use described in claim 4.

6. In the polypeptide, 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 defined by formula Connected to the base shown by, Forms the group shown by The use according to any one of claims 1 to 2.

7. In the polypeptide, 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 defined by formula Connected to the base shown by, Forms the group shown by The use according to any one of claims 1 to 2.

8. In the polypeptide, formula The structural units shown are, formula Selected from the group consisting of the groups shown. The use according to any one of claims 1 to 2.

9. The use of polypeptides having sequences selected from sequences represented by the following formula in the preparation of pharmaceuticals for the treatment and / or prevention of diabetes, obesity and related diseases, The structure of Aib is, It is a group represented by, The aforementioned diabetes-related diseases include diabetic cardiovascular disease, diabetic cerebrovascular disease, retinopathy, nephropathy, cataracts, coronary artery disease, diabetic neuropathy, diabetic foot disease, and peripheral neuropathy, and the aforementioned obesity-related diseases include liver disease, hyperlipidemia, and hypertension, as used in the foregoing.

10. The aforementioned medicines for treating and / or preventing diabetes, obesity and related diseases are administered at a frequency of once every three days, once every four days, once a week, or once every two weeks. The use according to any one of claims 1 to 2.

11. A pharmaceutical composition for treating and / or preventing diabetes mellitus, obesity and related diseases, comprising as an active ingredient a therapeutically effective amount of the compound for use described in any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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