Method for synthesizing amides and / or polypeptides using unprotected amino acids as amino components

Abstract

The present invention provides a method for synthesizing an amide and / or a polypeptide using an unprotected amino acid as an amino component. In the method for producing a polypeptide, a polypeptide is synthesized using an amino acid or a peptide segment in which neither the amino group nor the carboxyl group is protected as the amine component, and after an amide bond or a peptide bond is formed between an α-acyloxyamide derivative of a carboxylic acid and the amino group of the unprotected amino acid or peptide segment, the next cycle of constructing a peptide bond can be carried out without the need for a procedure to remove the carboxyl protecting group.

Description

[Technical Field]

[0001] The present invention belongs to the technical field of organic synthetic chemistry, and particularly relates to a method for synthesizing amides and / or polypeptides using unprotected amino acids as amino components. [Background technology]

[0002] Polypeptides and proteins are biological macromolecules with important functional activities, composed of naturally occurring amino acids linked in a specific order via amide bonds. They play an important regulatory role in various biological processes. Traditional methods for constructing peptide bonds primarily involve converting carboxylic acids into activated intermediates, such as acid chlorides, acid anhydrides, activated esters, or acyl azides, via activating or condensing agents, and then reacting them with the amines of other molecules to form peptide bonds. Among the various methods for forming peptide bonds, the condensing agent method is currently the most widely used in polypeptide synthesis. Hundreds of condensing agents have been developed, and the number of methods for synthesizing polypeptides is increasing. However, current polypeptide synthesis methods are still not ideal. For example, in the synthesis of dipeptides, the carboxyl group of one α-amino acid and the amino group of another α-amino acid must first be temporarily protected. After the carboxyl group of the protected α-amino acid forms a peptide bond with the amino group of another protected α-amino acid, the protecting groups on the amino or carboxyl groups must be removed before the next peptide bond can be constructed. Although the target polypeptide can be obtained by repeating this cycle of "protection," "condensation," and "deprotection," a large amount of protecting groups and deprotecting agents must be consumed throughout the polypeptide synthesis process, resulting in extremely low atom economy in the polypeptide synthesis process.

[0003] In 2016, a researcher designed and developed a new acetylene amide condensing agent for the synthesis of amide bonds and peptide bonds. This new condensing agent is easy to prepare, has good stability, a small molecular weight, mild reaction conditions, and requires no additives during use. More importantly, the α-chiral acid does not racemize during the condensation process, significantly improving the purity and yield of the product. However, this condensing agent still reacts with a carboxyl-protected amino acid as the amine component during polypeptide synthesis. After the reaction is complete, the resulting polypeptide is still protected, and deprotection is required to continue elongating the peptide chain. This makes the polypeptide synthesis process lengthy, time-consuming, and relatively expensive. Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to solve the problem of low atom economy due to the use of protecting groups in the above-mentioned conventional polypeptide synthesis techniques. [Means for solving the problem]

[0005] Therefore, the first aspect of the present invention proposes a method for producing amides and / or polypeptides. A polypeptide is synthesized using an amino acid or peptide segment in which neither the amino nor the carboxyl group is protected as an amine component. After an amide or peptide bond is formed between an α-acyloxyamide derivative of a carboxylic acid and the amino group of the unprotected amino acid or peptide segment, the next cycle of peptide bond construction can be carried out without the need for removing the carboxyl protecting group. Meanwhile, the acetylene amide condensing agent used in the present invention effectively avoids racemization. Previously, condensing agents or methods used in the synthesis of polypeptides using unprotected amino acids only produced dipeptides, but caused severe racemization when synthesizing tripeptides or polypeptides containing three or more amino acids. The method of the present invention avoids the racemization problem in the synthesis of polypeptides using unprotected amino acids. Furthermore, this method omits the steps of introducing protecting groups into amino acids and removing them from the product, thereby significantly reducing chemical usage and chemical waste generation, thereby making polypeptide synthesis more green and economical. This technology promotes a "dual-carbon" economy by saving energy and reducing pollutant emissions.

[0006] A second aspect of the present invention provides a method for producing an alternative amide and / or polypeptide.

[0007] A third aspect of the present invention proposes the use of a method for producing amides and / or polypeptides.

[0008] According to a first aspect of the present invention, there is provided a method for producing an amide and / or a polypeptide, said method comprising the steps of: [ka] reacting a compound of formula IV with a compound of formula V in a solvent II, and then subjecting a compound of formula II and an alkali additive to a nucleophilic substitution reaction in a solvent I to prepare a compound of formula III; where R 1is selected from an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue lacking a C-terminal carboxyl group, an amino acid derivative residue lacking a C-terminal carboxyl group, or a polypeptide fragment; R 2 is selected from hydrogen, deuterium, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a silyl group, or an alkylsilyl group; R 3 is selected from alkyl groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, and heteroaryl groups; R 4 is selected from an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue, or a polypeptide fragment; and EWG is selected from a nitro group, a cyano group, a sulfonyl group, an arylsulfonyl group, an alkanoyl group, or a phosphonoyl group, wherein the alkyl group, alkenyl group, alkynyl group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, silyl group, or alkylsilyl group is optionally unsubstituted or is selected from one or more R 11 Each R is replaced by 11 are independently selected from halogen, a hydroxy group, a carbonyl group, and a C1 to C6 alkyl group.

[0009] In some embodiments of the present invention, when solvent II is different from solvent I, the method for producing the amide and / or polypeptide further comprises the step of removing solvent II after the reaction and then performing the nucleophilic substitution reaction.

[0010] In some preferred embodiments of the present invention, the aryl group is selected from phenyl, o-tolyl, m-tolyl, p-tolyl, 2,4-xylyl, p-cumenyl, mesityl, 1-naphthyl, 2-naphthyl, 1-anthracenyl, 2-anthracenyl, 9-anthracenyl, 1-phenanthrenyl, 9-phenanthrenyl, 1-acenaphthylenyl, 2-acenaphthyl, 1-pyrenyl, 2-triphenylene, o-biphenyl, m-biphenyl, p-biphenyl, and terphenylyl.

[0011] In some more preferred embodiments of the present invention, the aromatic heterocycle is a heterocyclic group having one or two nitrogen atoms, oxygen atoms, or sulfur atoms and preferably having 5 to 10 members, such as a triazolyl group, a 3-oxadiazolyl group, a 2-furyl group, a 3-furyl group, a 2-thienyl group, a 3-thienyl group, a 1-pyrrolyl group, a 2-pyrrolyl group, a 3-pyrrolyl group, a 2-pyridyl group, a 3-pyridyl group, a 4-pyridyl group, a 2-pyrazinyl group, The alkyl group may be selected from, but is not limited to, 2-oxazolyl, 3-isoxazolyl, 2-thiazolyl, 3-isothiazolyl, 2-imidazolyl, 3-pyrazolyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl, 1-isoquinolyl, 2-quinoxalyl, 2-benzofuranyl, 2-benzothienyl, N-indolyl, and N-carbazolyl.

[0012] In some more preferred embodiments of the present invention, R 1 In the formula (I), the amino acid residue with a deleted C-terminal carboxyl group or amino acid derivative residue with a deleted C-terminal carboxyl group is the remaining portion of an amino acid or amino acid derivative after the C-terminal carboxyl group has been deleted. Preferably, the amino acid residue with a deleted C-terminal carboxyl group includes at least one of an α-amino acid residue with a deleted C-terminal carboxyl group, a β-amino acid residue with a deleted C-terminal carboxyl group, or a γ-amino acid residue with a deleted C-terminal carboxyl group. Preferably, the amino acid derivative residue with a deleted C-terminal carboxyl group includes an amino acid residue with a protected C-terminal amino group and a deleted C-terminal carboxyl group, for example, an amino acid residue with a deleted C-terminal carboxyl group and a protected C-terminal amino group with an N-alkoxycarbonyl group and / or an N-acyl group. Preferably, the amino acid residue is generally represented by the formula [ka] When the compound represented by R represents an amino acid, 1 wherein the amino acid residue lacking the C-terminal carboxyl group is represented by the formula [ka] is a group represented by the formula [ka] may be in the R or S configuration. Specifically, the amino acid residue lacking the C-terminal carboxyl group is of the formula [ka] Preferably, the amino acid derivative residue having a deleted C-terminal carboxyl group is selected from the above-mentioned amino acid residues having a deleted C-terminal carboxyl group, in which the C-terminal amino group is protected with an N-alkoxycarbonyl group and / or an N-acyl group.

[0013] According to a second aspect of the present invention, there is provided a method for producing an alternative amide and / or polypeptide, comprising: [ka] The method includes the steps of dissolving a compound of formula I, a compound of formula II, and an alkaline additive in a solvent I, and carrying out a nucleophilic substitution reaction to prepare a compound of formula III; where R 1 is selected from an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue lacking a C-terminal carboxyl group, an amino acid derivative residue lacking a C-terminal carboxyl group, or a polypeptide fragment; R 2 is selected from hydrogen, deuterium, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a silyl group, or an alkylsilyl group; R 3 is selected from alkyl groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, and heteroaryl groups; R 4is selected from an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue, or a polypeptide fragment; and EWG is selected from a nitro group, a cyano group, a sulfonyl group, an arylsulfonyl group, an alkanoyl group, or a phosphonoyl group, wherein the alkyl group, alkenyl group, alkynyl group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, silyl group, or alkylsilyl group is optionally unsubstituted or is selected from one or more R 11 Each R is replaced by 11 are independently selected from halogen, a hydroxy group, a carbonyl group, and a C1 to C6 alkyl group.

[0014] In the present invention, the compound of formula I is an activated ester, an α-acyloxyamide-based compound, whose ester carbonyl group has relatively good electrophilic activity and can undergo nucleophilic substitution with a nucleophilic reagent. Using an alkali additive, the amino group of the ammonium ion, which has no or very weak nucleophilicity, in the compound of formula II-A (a compound containing both an amino group and a carboxyl group) is liberated to form the compound of formula IA. The compound of formula II-A can undergo nucleophilic substitution with the compound of formula I to form an amide bond / peptide bond without protecting the carboxyl group, producing the compound of formula III, which is an amide or polypeptide, and also forming the by-product compound of formula IV. The reaction mechanism is as follows:

[0015] [ka]

[0016] In some embodiments of the present invention, the alkaline additive comprises an organic base and / or an inorganic base, preferably the organic base is selected from the group consisting of trimethylamine, triethylamine, tripropylamine, N,N-dimethylethylamine, N,N-diethylmethylamine, N,N-diisopropylethylamine, N-methylpyrrolidine, N-methylpiperidine, N-methylmorpholine, tributylamine, tribenzylamine, dimethylbenzylamine, N,N-dimethylaniline, pyridine, 2-methylpyridine, 2,6-dimethylpyridine, 4-dimethylaminopyridine, quinoline, quinine, tetramethylguanidine, imidazole, 1,8-diazabicyclo[5.4.0]undec-7-ene, the inorganic base comprises at least one of sodium bicarbonate, sodium carbonate, lithium bicarbonate, lithium carbonate, potassium bicarbonate, potassium carbonate, cesium bicarbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium hydroxide, calcium bicarbonate, or calcium hydroxide.

[0017] In some preferred embodiments of the present invention, the molar ratio of the alkaline additive to the compound of formula II is (0.5-20):1, preferably (1-15):1, and more preferably (2-10):1.

[0018] In some preferred embodiments of the present invention, the compound of formula II is an organic compound having both an amino group and a carboxyl group. Preferably, the compound of formula II includes any one of amino acid compounds, polypeptide compounds in which both the amino group and the carboxyl group are free, or other compounds having both an amino group and a carboxyl group. More preferably, the compound of formula II includes any one of α-amino acids, β-amino acids, γ-amino acids, or polypeptide compounds in which neither the amino group nor the carboxyl group is protected. Even more preferably, the compound of formula II includes any one of glycine, alanine, leucine, isoleucine, proline, valine, phenylalanine, tyrosine, serine, threonine, cysteine, methionine, aspartic acid, glutamic acid, asparagine, glutamine, tryptophan, histidine, lysine, arginine, citrulline, or derivatives of the above amino acids. In addition, the 21 amino acids contained in the compound of formula II may be natural α-amino acids in their L-configuration and / or the corresponding α-amino acids in their D-configuration.

[0019] In some more preferred embodiments of the invention, the solvent I comprises at least one of water, acetonitrile, ethanol, isopropyl alcohol, tert-butanol, ethylene glycol, 1,2-propanediol, 2,3-butanediol, ethyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, toluene, m-xylene, dichloromethane, 1,2-dichloroethane, or chloroform.

[0020] In some more preferred embodiments of the present invention, the temperature of the nucleophilic substitution reaction is -20°C to 150°C, preferably 0°C to 90°C, and more preferably 5°C to 50°C.

[0021] In some more preferred embodiments of the present invention, the method for producing the amide and / or polypeptide comprises dissolving a compound of formula II and an alkaline additive in solvent I, and adding a compound of formula I to carry out a nucleophilic substitution reaction to produce a compound of formula III.

[0022] In some more preferred embodiments of the present invention, the method for producing the amide and / or polypeptide includes the steps of dissolving a compound of formula II and an alkaline additive in solvent I by stirring for 0.5 min to 12 hours, adding a compound of formula I, and carrying out a nucleophilic substitution reaction to produce a compound of formula III.

[0023] In some more preferred embodiments of the present invention, the method for producing the amide and / or polypeptide further comprises purifying the product after the nucleophilic substitution reaction, wherein the purification comprises at least one of extraction, recrystallization, or column chromatography.

[0024] In some more preferred embodiments of the present invention, the method for preparing compounds of formula I comprises: [ka] dissolving a compound of formula IV and a compound of formula V in a solvent II, and stirring and reacting them to produce a compound of formula I; where R 1 , R 2 , R 3 The definition of EWG is as described above.

[0025] In some more preferred embodiments of the present invention, the solvent II comprises at least one of dichloromethane, chloroform, 1,2-dichloroethane, ethyl ether, toluene, acetonitrile, methanol, or ethanol.

[0026] In some more preferred embodiments of the present invention, the compound of formula IV is a carboxylic acid-based compound, preferably the compound of formula IV comprises at least one of a fatty acid, an aromatic acid, a heterocyclic acid, an alkynoic acid, an olefinic acid, an N-acyl amino acid, an N-alkoxycarbonyl amino acid, or a polypeptide carboxylic acid, more preferably the compound of formula IV comprises at least one of an N-benzyloxycarbonyl amino acid, an N-tert-butoxycarbonyl amino acid, an N-fluorenemethoxycarbonyl amino acid, an N-acetyl amino acid, or a polypeptide carboxylic acid.

[0027] In some more preferred embodiments of the present invention, the molar ratio of the compound of formula IV to the compound of formula V is 1:(1-5), preferably 1:(1.1-4), and more preferably 1:(1.2-3).

[0028] In some more preferred embodiments of the present invention, the temperature of the stirring reaction is -20°C to 90°C, preferably 0°C to 50°C.

[0029] In some more preferred embodiments of the present invention, the method for producing the amide and / or polypeptide comprises: [ka] Step S1: dissolving the compound of formula IV and the compound of formula V in a solvent II, stirring and reacting them to prepare a compound of formula I; Step S2 of dissolving the compound of formula I, the compound of formula II and an alkaline additive in a solvent I and carrying out a nucleophilic substitution reaction to prepare a compound of formula III; where R 1 , R 2 , R 3 , R 4 The definition of EWG is as described above.

[0030] According to a third aspect of the present invention, there is provided a method for producing an amide and / or a polypeptide, the method being used in the production of a polypeptide. [Effects of the Invention]

[0031] The beneficial effects of the present invention are as follows:

[0032] In the method for producing amides and / or polypeptides of the present invention, low-cost unprotected amino acids are used as starting materials, eliminating the need to introduce a protecting group into the carboxyl group of an amino acid, and avoiding the need to remove the protecting group after constructing a peptide bond, allowing for direct sequential condensation with the amino group of the next amino acid. This method saves on the introduction of protecting group reagents, deprotection reagents, and solvent usage, reduces the generation of large amounts of chemical waste, saves manpower and energy consumption, significantly reduces the cost of polypeptide synthesis, improves the atom economy and step economy of polypeptide synthesis, effectively achieves energy conservation and reduces pollutant emissions, greatly improves the efficiency of polypeptide synthesis, and has broad industrial potential. DETAILED DESCRIPTION OF THE INVENTION

[0033] In order to fully understand the objectives, features and advantages of the present invention, the concept and technical advantages of the present invention will be clearly and completely described in the following embodiments. Obviously, the described embodiments are only some of the embodiments of the present invention, not all of the embodiments, and any other embodiments obtained by those skilled in the art based on the embodiments of the present invention without any creative efforts are also within the scope of protection of the present invention. [Example]

[0034] Example 1 In this example, the polypeptide of formula 1 is prepared, and the specific steps are as follows:

[0035] [ka]

[0036] Cbz-Gly-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure. H-Ala-OH (1 mmol), triethylamine (1 mmol), 3 mL of N,N-dimethylformamide, and 1 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 2 with 2 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, washed once with saturated brine, and dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure. The product was recrystallized from ethyl ether, and the solid was collected by filtration to obtain the pure product as a white solid in 95% yield.

[0037] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO)δ12.63(s,1H),8.15(d,J=7.2Hz,1H),7.43(t,J=5.9Hz,1H),7.38- 7.30(m,5H),5.04(s,2H),4.29-4.19(m,1H),3.71-3.62(m,2H),1.28(d,J=7.2Hz,3H). 13 C NMR(100MHz,DMSO)δ=174.1,168.9,156.5,137.1,128.4,127.8,127.7,65.5,47.5,43.3,17.4. HRMS(ESI) m / z calculation for C 13 H 16 N2NaO5[M+Na] + :303.0951,found:303.0950

[0038] Example 2 In this example, the polypeptide of formula 2 is prepared, and the specific steps are as follows:

[0039] [ka]

[0040] Cbz-Leu-OH (0.5 mmol) and N-methyl-N-methanesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After complete consumption of the starting acid, the dichloromethane was removed by distillation under reduced pressure. H-Phe-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 2 with 2 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The pure product was obtained by column chromatography as a white solid with a DR of >99:1 and a yield of 97%.

[0041] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,MeOD-d4)δ7.36-7.26(m,5H),7.24-7.14(m,5H),5.06(s,2H),4.69(dd,J=8.0,5.3Hz,1H),4.26-4.14(m,1 H),3.18(dd,J=13.8,5.1Hz,1H),3.00(dd,J=13.8,8.1Hz,1H),1.67-1.59(m,1H),1.52-1.41(m,2H),0.96-0.85(m,6H). 13 C NMR(100MHz,MeOD-d4)δ174.9,174.3,158.2,138.0,138.0,130.4,129.4,129.3,128.9,128.7,127.7,67.6,54.7,42.0,38.3,25.7,23.4,22.0. HRMS(ESI) m / z calculation for C 23 H 28 N2NaO5[M+Na] +:435.1890,found:435.1883

[0042] Example 3 In this example, the polypeptide of formula 3 is prepared, and the specific steps are as follows:

[0043] [ka]

[0044] Cbz-Ser(tBu)-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.5 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure. H-Leu-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), 3 mL of dimethyl sulfoxide, and 1.5 mL of water were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and washed once with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, and the mixture was separated by column chromatography. The solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 93%.

[0045] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ7.98(d,J=7.9Hz,1H),7.38-7.28(m,5H),7.21(d,J=8.5Hz,1H),5.04(s,2H),4.29-4.23(m,1H) ,4.18-4.12(m,1H),3.50-3.43(m,2H),1.70-1.62(m,1H),1.56-1.47(m,2H),1.10(s,9H),0.86(dd,J=16.6,6.3Hz,6H). 13C NMR(100MHz,DMSO-d6)δ173.9,169.7,155.8,137.0,128.3,127.8,127.7,72.8,65.5,62.0,55.4,50.3,40.3,27.2,24.1,22.9,21.4. HRMS(ESI) m / z calculation for C 21 H 32 N2NaO6[M+Na] + :431.2153,found:431.2149

[0046] Example 4 In this example, the polypeptide of formula 4 is prepared, and the specific steps are as follows:

[0047] [ka]

[0048] Cbz-Thr-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure. H-Leu-OH (1 mmol), sodium carbonate (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 2 with 2 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, the solvent was removed by distillation under reduced pressure, and the product was recrystallized from ethyl ether. The solid was collected by filtration to obtain the pure product, a white solid with a DR>99:1 ratio and a yield of 94%.

[0049] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ8.06(d,J=7.7Hz,1H),7.41-7.17(m,10H),6.95(d,J=8.7Hz,1H),5.05(s,2H),4.48(q,J=7.3Hz,1H),3. 96(dd,J=8.2,5.2Hz,1H),3.88-3.76(m,1H),3.05(dd,J=13.7,5.0Hz,1H),2.93(dd,J=13.6,8.1Hz,1H),1.01(d,J=6.1Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ172.6,170.0,156.0,137.3,137.0,129.2,128.3,128.1,127.8,127.7,126.4,66.8,65.5,60.5,53.3,36.9,19.6. HRMS(ESI) m / z calculation for C 21 H 24 N2NaO6[M+Na] + :423.1527,found:423.1524

[0050] Example 5 In this example, the polypeptide of formula 5 is prepared, and the specific steps are as follows:

[0051] [ka]

[0052] Boc-Met-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.5 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After complete consumption of the starting acid, the dichloromethane was removed by distillation under reduced pressure. H-Leu-OH (1 mmol), triethylamine (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The pure product was obtained by column chromatography as a white solid with a pH of >99:1 and a yield of 94%.

[0053] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ7.97(d,J=7.8Hz,1H),7.34-7.16(m,5H),6.91(d,J=8.2Hz,1H),4.46(q,J=8.0Hz,1H),4.01(q,J=7.9Hz, 1H),3.07(dd,J=13.8,4.9Hz,1H),2.92(dd,J=13.9,8.7Hz,1H),2.45-2.29(m,2H),2.01(s,3H),1.85-1.64(m,2H),1.37(s,9H). 13 C NMR(100MHz,DMSO-d6)δ172.8,171.6,155.2,137.3,129.2,128.1,126.4,78.2,53.6,53.2,36.7,31.9,29.6,28.2,14.6. HRMS(ESI) m / z calculation for C 19 H 28 N2NaO5S [M+Na] + :419.1611,found:419.1609

[0054] Example 6 In this example, the polypeptide of formula 6 is prepared, and the specific steps are as follows:

[0055] [ka]

[0056] Boc-Ala-OH (0.5 mmol) and N-methyl-N-methanesulfonylethinamine (0.5 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After complete consumption of the starting acid, the dichloromethane was removed by distillation under reduced pressure. H-Lys(Cbz)-OH (1 mmol), potassium carbonate (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The pure product was obtained by column chromatography as a white solid with a pH of >99:1 and a yield of 94%.

[0057] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ7.88(d,J=7.8Hz,1H),7.39-7.27(m,5H),7.20(t,J=5.8Hz,1H),6.86(d,J=7.7Hz,1H),5.00(s,2H),4.20-4.13(m ,1H),4.03-3.96(m,1H),3.01-2.94(m,2H),1.75-1.65(m,1H),1.63-1.53(m,1H),1.37(s,9H),1.34-1.26(m,3H),1.17(d,J=7.1Hz,3H). 13C NMR(100MHz,DMSO-d6)δ173.6,172.8,156.1,155.1,137.3,128.4,127.8,127.7,78.1,65.2,51.7,49.6,40.1,30.9,29.0,28.2,22.6,18.1. HRMS(ESI) m / z calculation for C 22 H 34 N2O7[M+H] + :452.2391,found:452.2390

[0058] Example 7 In this example, the polypeptide of formula 7 is prepared, and the specific steps are as follows:

[0059] [ka]

[0060] Cbz-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After complete consumption of the starting acid, the dichloromethane was removed by distillation under reduced pressure. H-Thr-OH (1 mmol), sodium bicarbonate (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 2 with 2 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, the solvent was removed by distillation under reduced pressure, and the product was recrystallized with ethyl acetate and ethyl ether. The solid was collected by filtration to obtain the pure product, a white solid with a DR>99:1 ratio and a yield of 92%.

[0061] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ7.67(d,J=8.6Hz,1H),7.53(d,J=7.7Hz,1H),7.39-7.26(m ,5H),5.03(s,2H),4.25-4.14(m,3H),1.24(d,J=7.1Hz,3H),1.06(d,J=6.1Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ173.0,172.2,155.8,137.1,128.5,127.9,127.8,66.5,65.5,57.5,50.2,20.4,18.2. HRMS(ESI) m / z calculation for C 15 H 20 N2O6[M+Na] + :347.1214,found:347.1221

[0062] Example 8 In this example, the polypeptide of formula 8 is prepared, and the specific steps are as follows:

[0063] [ka]

[0064] Cbz-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure. H-Gln-OH (1 mmol) was added, and 3 mL of acetonitrile and 1.5 mL of water were added as solvents. N,N-diisopropylethylamine (1 mmol) was added, and the reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=1 with 2 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The product was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 97%.

[0065] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.13(d,J=7.6Hz,1H),7.59-7.12(m,7H),6.79(s,1H),5.09-4.95(m,2H), 4.22-4.05(m,2H),2.22-2.07(m,2H),2.01-1.91(m,1H),1.86-1.73(m,1H),1.22(d,J=7.1Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ173.6,173.3,172.7,155.7,137.1,128.4,127.8,127.8,65.4,51.6,49.9,31.4,27.0,18.2. HRMS(ESI) m / z calculation for C 16 H 21 N3NaO6[M+Na] + :374.1323,found:374.1321

[0066] Example 9 In this example, the polypeptide of formula 9 is prepared, and the specific steps are as follows:

[0067] [ka]

[0068] Cbz-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After complete consumption of the starting acid, the dichloromethane was removed by distillation under reduced pressure. H-Trp-OH (1 mmol), cesium carbonate (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 1 with 2 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, the solvent was removed by distillation under reduced pressure, and the pure product was obtained by column chromatography as a pale yellow solid with a pH of >99:1 and a yield of 93%.

[0069] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ10.89(s,1H),8.12-8.03(m,1H),7.57(t,J=6.0Hz,1H),7.44(dd,J=8.1,4.1Hz,1H),7.34(d,J=15.7Hz,8H),7.20(s,1H),7 .07(s,1H),6.98(s,1H),5.13-4.94(m,3H),4.54(s,1H),4.15(q,J=5.8, 4.9Hz,2H),3.22(d,J=14.3Hz,2H),3.17-3.05(m,2H),1.26-1.19(m,7H). 13 C NMR(100MHz,DMSO-d6)δ173.3,172.6,155.7,137.1,136.1,128.4,127.8,127.8 ,127.4,123.8,121.0,118.4,118.3,111.4,109.7,65.5,53.0,50.0,27.1,18.3. HRMS(ESI) m / z calculation for C 22 H 23 N3NaO5[M+Na] + :432.1530,found:432.1530

[0070] Example 10 In this example, the polypeptide of formula 10 is prepared, and the specific steps are as follows:

[0071] [ka]

[0072] Fmoc-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After complete consumption of the starting acid, the dichloromethane was removed by distillation under reduced pressure. H-Phe-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 1 with 2 M hydrochloric acid, and the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were washed once with water and once with saturated brine. The organic phase was dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure. The pure product was obtained by column chromatography as a white solid with a pH of >99:1 and a yield of 91%.

[0073] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.02(d,J=7.5Hz,1H),7.89(d,J=7.5Hz,2H),7.74(d,J=6.8H z,1H),7.48(d,J=7.7Hz,1H),7.42(t,J=7.4Hz,2H),7.33(t,J=7.3Hz,2H),7.27-7.16 (m,4H),4.50-4.39(m,1H),4.26(d,J=6.1Hz,1H),4.24-4.18(m,1H),4.12-4.06(m,1 H),3.07(dd,J=13.7,4.8Hz,1H),2.93(dd,J=13.7,8.4Hz,1H),1.20(d,J=7.1Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ172.8,172.4,155.6,143.9,143.8,140.7,137.4,129.2 ,128.1,127.6,127.1,126.3,125.3,120.1,65.7,53.5,49.9,46.6,36.7,18.2. HRMS(ESI) m / z calculation for C 22 H 23 N3NaO5[M+Na] + :432.1530,found:432.1530

[0074] Example 11 In this example, the polypeptide of formula 11 is prepared, and the specific steps are as follows:

[0075] [ka]

[0076] Cbz-Ala-Leu-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.5 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Phe-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), and 5 mL of ethylene glycol were added to the pre-mixed mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 1 with 2 M hydrochloric acid, the aqueous phase was extracted twice with dichloromethane, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, the solvent was removed by distillation under reduced pressure, and the mixture was recrystallized with ethyl ether. The solid was collected by filtration to obtain the pure product, a white solid with a pH of >99:1 and a yield of 93%.

[0077] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ8.04(d,J=7.6Hz,1H),7.86(d,J=8.3Hz,1H),7.43(d,J=7 .6Hz,1H),7.37-7.17(m,10H),5.02(s,2H),4.47-4.41(m,1H),4.36-4.30(m,1H) ,4.10-4.04(m,1H),3.07(dd,J=13.9,5.2Hz,1H),2.93(dd,J=13.8,8.5Hz,1H),1 .67-1.50(m,1H),1.43(d,J=6.7Hz,1H),1.17(d,J=7.0Hz,3H),0.90-0.76(m,6H). 13 C NMR(100MHz,DMSO-d6)δ172.8,172.2,171.9,155.7,137.5,137.1,129.2,128.4,128 .2,127.8,127.8,126.4,65.4,53.4,51.0,50.1,41.1,36.7,24.1,23.1,21.8,18.2. HRMS(ESI) m / z calculation for C 26 H 33 N3NaO6[M+Na] + :506.2262,found:506.2253

[0078] Example 12 In this example, the polypeptide of formula 12 is prepared, and the specific steps are as follows:

[0079] [ka]

[0080] Cbz-Ala-Phe-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.5 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Met-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), and 5 mL of 2,3-butanediol were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 1 with 2 M hydrochloric acid, the aqueous phase was extracted twice with dichloromethane, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The product was recrystallized from dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a DR>99:1 ratio and a yield of 93%.

[0081] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.18(d,J=7.3Hz,1H),7.94(d,J=7.7Hz,1H),7.43(d,J= 7.0Hz,1H),7.40-7.12(m,10H),5.10-4.93(m,2H),4.60-4.51(m,1H),4.41-4.3 4(m,1H),4.05-3.98(m,1H),3.14-3.03(m,1H),2.91-2.79(m,1H),2.52-2.42(m ,2H),2.04(s,3H),2.02-1.95(m,1H),1.93-1.85(m,1H),1.13(d,J=7.1Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ173.2,172.3,171.0,155.8,137.7,137.0,129.4,128.4, 128.0,127.9,127.8,126.3,65.6,53.6,51.1,50.4,37.3,30.9,29.7,18.1,14.7. HRMS(ESI)m / z calcd. for C 25 H 31 N3NaO6S [M+Na] +:524.1826,found:524.1824

[0082] Example 13 In this example, the polypeptide of formula 13 is prepared, and the specific steps are as follows:

[0083] [ka]

[0084] Cbz-Ala-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Gln(Trt)-OH (1 mmol), sodium bicarbonate (2 mmol), and 5 mL of dichloromethane were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted once with ethyl acetate, washed twice with water, and the organic phase was washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The product was recrystallized from dichloromethane and ethyl ether. The solid was collected by filtration to obtain the pure product, a pale yellow solid with a pH of >99:1 and a yield of 90%.

[0085] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.59(s,1H),8.05(d,J=8.2Hz,1H),7.93(t,J=6.5Hz,1H),7.44(t,J=5.9Hz,1H),7.38-7.14(m,22H),5.06-4.97(m ,2H),4.37-4.29(m,1H),4.22-4.15(m,1H),4.11-4.05(m,1H),2.41- 2.29(m,2H),1.99-1.88(m,1H),1.79-1.69(m,1H),1.26-1.16(m,6H). 13C NMR(100MHz,DMSO-d6)δ173.3,172.1,172.0,171.2,155.7,144.9,137.0,128.5,128 .3,127.8,127.7,127.4,126.3,69.3,65.4,51.6,50.0,47.9,32.6,27.3,18.3,18.1. HRMS(ESI) m / z calculation for C 38 H 40 N4NaO7[M+Na] + :687.2789,found:687.2874

[0086] Example 14 In this example, the polypeptide of formula 14 is prepared, and the specific steps are as follows:

[0087] [ka]

[0088] Cbz-Phe-Gly-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure. H-Phe-Ser(tBu)-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 92%.

[0089] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ8.33-8.17(m,2H),8.05(d,J=8.4Hz,1H),7.52(d,J=8.5Hz,1H),7.38-7.13(m,15H),5.02-4.87(m,2H),4.77-4.65(m,1H), 4.43-4.35(m,1H),4.32-4.25(m,1H),3.80-3.74(m,1H),3.69-3.60(m,2 H),3.57-3.52(m,1H),3.11-2.98(m,2H),2.83-2.68(m,2H),1.13(s,9H). 13 C NMR(100MHz,DMSO-d6)δ171.8,171.6,171.1,168.4,155.9,138.2,137.8,137.0,129.3,129.2,128.3, 128.2,128.1,128.0,127.7,127.4,126.3,72.9,65.3,61.6,56.2,53.6,53.1,41.9,37.8,37.5,27.2. HRMS(ESI) m / z calculation for C 35 H 42 N4NaO8[M+Na] + :669.2895,found:669.2900

[0090] Example 15 In this example, the polypeptide of formula 15 is prepared, and the specific steps are as follows:

[0091] [ka]

[0092] Cbz-Phe-Gly-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Glu(tBu)-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), and 5 mL of 2,3-butanediol were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 89%.

[0093] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.38-8.28(m,1H),8.14(d,J=6.9Hz,1H),7.96(d,J=6 .0Hz,1H),7.57(d,J=7.6Hz,1H),7.35-7.15(m,10H),4.95(s,2H),4.41-4.18 (m,3H),3.82-3.71(m,2H),3.10-3.01(m,1H),2.83-2.71(m,1H),2.36-2.18( m,2H),2.05-1.90(m,1H),1.88-1.72(m,1H),1.38(s,9H),1.27-1.19(m,3H). 13 C NMR(100MHz,DMSO-d6)δ173.1,172.2,171.9,171.5,168.2,156.0,138.1,137.0,129.2,128.3 ,128.0,127.7,127.4,126.2,79.8,65.3,56.3,51.1,47.9,42.1,37.3,31.3,27.7,26.4,18.3. HRMS(ESI) m / z calculation for C 31 H 41N4O9[M+H] + :613.2868,found:613.2865

[0094] Example 16 In this example, the polypeptide of formula 16 is prepared, and the specific steps are as follows:

[0095] [ka]

[0096] Cbz-Tyr(tBu)-Gly-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure. H-Gly-Phe-OH (1 mmol), sodium carbonate (1 mmol), 3 mL of acetonitrile, and 1.5 mL of water were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 96%.

[0097] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ8.38-8.26(m,1H),8.18(d,J=6.6Hz,1H),8.11-7.96(m,1H),7.64-7.43(m,1H),7.36-7.15(m,12H),6.86(d,J=7.4Hz,2H), 5.01-4.86(m,2H),4.53-4.43(m,1H),4.35-4.26(m,1H),3.84-3.70(m,4 H),3.12-3.00(m,2H),2.97-2.88(m,1H),2.80-2.69(m,1H),1.26(s,9H). 13 C NMR(100MHz,DMSO-d6)δ172.8,172.0,169.0,168.6,156.0,153.5,137.5,137.0,132.8,129.8,129 .2,128.3,128.3,127.7,127.5,126.5,123.3,77.6,65.4,56.3,53.6,42.2,41.7,36.9,36.8,28.6.

[0098] Example 17 In this example, the polypeptide of formula 17 is prepared, and the specific steps are as follows:

[0099] [ka]

[0100] Cbz-Gly-Ala-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Ala-Phe-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), and 5 mL of 2,3-butanediol were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 89%.

[0101] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.00(t,J=6.8Hz,3H),7.51-7.41(m,1H),7.42-7.16(m,10H),5.04(s,2H),4.47-4.38(m,1H),4.31 (p,J=7.2Hz,2H),3.66(d,J=5.9Hz,2H),3.06(dd,J=13.8,5.1Hz,1H),2.93(dd,J=13.6,8.5Hz,1H),1.19(t,J=8.1Hz,6H). 13 C NMR(100MHz,DMSO-d6)δ172.7,172.0,171.7,168.8,156.5,137.4,137.1,129. 2,128.4,128.2,127.8,127.7,126.5,65.5,53.5,48.0,43.5,36.7,18.4,18.1. HRMS(ESI) m / z calculation for C 25 H 30 N4NaO7[M+Na] + :521.2007,found:521.2009

[0102] Example 18 In this example, the polypeptide of formula 18 is prepared, and the specific steps are as follows:

[0103] [ka]

[0104] Cbz-Ala-Ser(tBu)-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Phe-OH (1 mmol), tributylamine (1 mmol), and 5 mL of dichloromethane were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The product was recrystallized from dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a DR>99:1 ratio and a yield of 95%.

[0105] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ7.97(d,J=7.8Hz,1H),7.74(d,J=7.7Hz,1H),7.49(d,J=7.3Hz,1H),7.39-7.14(m,10H),5.02(s,2H),4.53-4.44( m,1H),4.38-4.30(m,1H),4.16-4.07(m,1H),3.49-3.38(m,2H),3.10-3.03(m,1H),2.97-2.88(m,1H),1.19(d,J=6.9Hz,3H),1.09(s,9H). 13C NMR(101MHz,DMSO)δ=172.4,172.3,169.5,155.7,137.3,137.0,129.2,128.3,1 28.2,127.8,127.7,126.4,72.9,65.4,61.7,53.3,53.0,50.1,36.9,27.1,18.1. HRMS(ESI) m / z calculation for C 27 H 35 N3NaO7[M+Na] + :536.2367,found:536.2372

[0106] Example 19 In this example, the polypeptide of formula 19 is prepared, and the specific steps are as follows:

[0107] [ka]

[0108] Cbz-Ala-Cys(Trt)-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Phe-OH (1 mmol), N-methylmorpholine (0.5 mmol), triethylamine (1 mmol), and 5 mL of dichloromethane were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 95%.

[0109] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ8.18-7.99(m,2H),7.56(d,J=7.4Hz,1H),7.36-7.14(m,25H),5.12-4.91(m,2H),4.46- 4.32(m,2H),4.14-4.04(m,1H),3.09-2.99(m,1H),2.95-2.85(m,1H),2.44-2.30(m,2H),1.19(d,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO)δ=172.4,172.3,169.5,155.8,144.3,137.2,137.0,129.1,129.1,128.4 ,128.2,128.1,127.8,127.8,126.8,126.5,65.8,65.5,53.5,51.6,50.2,36.7,33.8,18.2. HRMS(ESI) m / z calculation for C 42 H 41 N3NaO6S [M+Na] + :738.2608,found:738.2615

[0110] Example 20 In this example, the polypeptide of formula 20 is prepared, and the specific steps are as follows:

[0111] [ka]

[0112] Cbz-Ala-Glu(tBu)-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Asp(tBu)-OH (1 mmol), N,N-dimethylaniline (0.5 mmol), triethylamine (1 mmol), and 5 mL of dichloromethane were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 97%.

[0113] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.19(d,J=7.9Hz,1H),7.98(d,J=8.1Hz,1H),7.47(d,J=7 .5Hz,1H),7.39-7.28(m,5H),5.07-4.99(m,2H),4.60-4.52(m,1H),4.37-4.28(m ,1H),4.14-4.05(m,1H),2.74-2.66(m,1H),2.61-2.52(m,1H),2.31-2.16(m,2H) ,1.98-1.83(m,1H),1.81-1.63(m,1H),1.42-1.34(m,18H),1.21(d,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO)δ=172.4,172.1,171.8,170.8,169.2,155.7,137.0,128.4,127 .8,127.8,80.4,79.6,65.4,51.4,50.1,48.7,37.2,31.1,27.8,27.8,27.7,18.1. HRMS(ESI) m / z calculation for C 28 H 41N3NaO 10 [M+Na] + :602.2684,found:602.2690.

[0114] Example 21 In this example, the polypeptide of formula 21 is prepared, and the specific steps are as follows:

[0115] [ka]

[0116] Cbz-Ala-Glu(tBu)-Asp(tBu)-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Gly-OH (1 mmol), 2,6-dimethylpyridine (2 mmol), and 5 mL of dichloromethane were added to the previously stirred mixture. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a dr>99:1 and a yield of 93%.

[0117] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1H NMR(400MHz,DMSO-d6)δ8.14(d,J=8.0Hz,1H),8.10-7.95(m,2H),7.47(d,J=7.2Hz,1H ),7.40-7.22(m,5H),5.10-4.94(m,2H),4.67-4.55(m,1H),4.30-4.21(m,1H),4.13-4. 04(m,1H),3.82-3.67(m,2H),2.76-2.62(m,1H),2.50-2.36(m,1H),2.32-2.10(m,2H), 2.02-1.82(m,1H),1.82-1.62(m,1H),1.39(s,9H),1.37(s,9H),1.20(d,J=6.9Hz,3H). 13 C NMR(101MHz,DMSO)δ=172.8,171.8,170.9,170.8,170.4,169.2,155.8,137.0,128.3,12 7.8,127.8,80.2,79.6,65.5,52.0,50.1,49.4,40.9,37.4,31.1,27.8,27.7,27.3,18.0. HRMS(ESI) m / z calculation for C 30 H 44 N4NaO 11 [M+Na] + :659.2899,found:659.2905.

[0118] Example 22 In this example, the polypeptide of formula 22 is prepared, and the specific steps are as follows:

[0119] [ka]

[0120] Cbz-Ala-Phe-Met-OH (0.5 mmol) and N-methyl-N-toluenesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, H-Glu(tBu)-OH (1 mmol), pyridine (2 mmol), and 5 mL of dichloromethane were added to the pre-stirred mixture. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed once with water, the organic phase was washed once with saturated brine, the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The product was recrystallized from dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product, a white solid with a DR>99:1 ratio and a yield of 92%.

[0121] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.13(d,J=7.5Hz,1H),8.07(d,J=7.4Hz,1H),7.93(d,J=7.3Hz,1H),7.43(d,J= 7.1Hz,1H),7.40-7.28(m,5H),7.25-7.15(m,5H),5.09-4.93(m,2H),4.60-4.49(m,1H),4.44-4.36(m, 1H),4.28-4.20(m,1H),4.05-3.98(m,1H),3.09-3.00(m,1H),2.90-2.80(m,1H),2.49-2.40(m,2H),2. 33-2.23(m,2H),2.03(s,3H),2.02-1.89(m,2H),1.88-1.74(m,2H),1.39(s,9H),1.14(d,J=7.0Hz,3H). 13C NMR(101MHz,DMSO)δ=173.0,172.4,171.5,170.9,170.7,155.7,137.6,136.9,129.3,128.3,127.9,12 7.8,127.7,126.2,79.8,65.5,53.6,51.7,51.1,50.3,37.2,32.2,31.2,29.3,27.7,26.3,18.0,14.6. HRMS(ESI) m / z calculation for C 34 H 46 N4NaO9S [M+Na] + :709.2878,found:709.2884.

[0122] Example 23 In this example, the amide of formula 23 is prepared by the following steps:

[0123] [ka]

[0124] 18-(tert-butoxy)-18-oxooctadecanoic acid (0.5 mmol) and N-methyl-N-methanesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the solvent was removed by distillation under reduced pressure. The remaining mixture was transferred to a pre-stirred mixture of H-Glu-OtBu (1 mmol), N,N-diisopropylethylamine (1 mmol), 3 mL of dimethyl sulfoxide, and 3 mL of water. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product as a white solid in a yield of 95%.

[0125] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.00(d,J=7.6Hz,1H),4.16-4.09(m,1H),2.29-2.23(m,2H),2.12(dt,J=21.7,7.3Hz,4H),1. 90(dt,J=13.5,6.6Hz,1H),1.75(dd,J=14.1,6.7Hz,1H),1.52-1.44(m,4H),1.41-1.37(m,18H),1.27-1.21(m,24H). 13 C NMR(101MHz,DMSO)δ173.53,172.25,172.08,171.03,80.26,79.16,79.14,51.81,34.97,34.74,2 9.96,29.00,28.96,28.91,28.82,28.76,28.60,28.53,28.33,27.68,27.55,26.24,25.20,24.55. HRMS(ESI) m / z calculation for C 31 H 57 NNaO7[M+Na] + :578.4027,found:578.4034.

[0126] Example 24 In this example, the amide of formula 24 is prepared by the following steps:

[0127] [ka]

[0128] Fmoc-β-Ala-OH (0.5 mmol) and N-methyl-N-methanesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the solvent was removed by distillation under reduced pressure. The remaining mixture was transferred to a pre-mixed system of H-Phe-OH (1 mmol), N,N-diisopropylethylamine (1 mmol), 3 mL of acetonitrile, and 3 mL of water. The reaction progress was monitored by TLC. After the reaction was completed, 15 mL of water was added, the mixture was acidified to pH=3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. The mixture was recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product as a white solid in a yield of 90%.

[0129] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ8.24(d,J=7.6Hz,1H),7.88(d,J=7.5Hz,2H),7.69(d ,J=7.2Hz,2H),7.41(t,J=7.4Hz,2H),7.33(t,J=7.4Hz,2H),7.29-7.13(m,6 H),4.53-4.42(m,1H),4.29(d,J=6.5Hz,2H),4.22(d,J=6.4Hz,1H),3.21-3. 05(m,3H),2.89(dd,J=13.2,9.4Hz,1H),2.30(dq,J=19.6,13.0,10.2Hz,2H). 13 C NMR(101MHz,DMSO)δ173.05,170.26,155.99,143.90,140.72,137.67,129.06,128.12 ,127.59,127.05,126.36,125.15,120.07,65.39,53.43,46.75,36.98,36.84,35.40. HRMS(ESI) m / z calculation for C 27 H 26 N2NaO5[M+Na]+ :481.1734,found:481.1739.

[0130] Example 25 In this example, the amide of formula 25 is prepared by the following steps:

[0131] [ka]

[0132] Boc-Phe-OH (0.5 mmol) and N-methyl-N-methanesulfonylethinamine (0.55 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the solvent was removed by distillation under reduced pressure. The remaining mixture was transferred to a pre-stirred mixture of γ-aminobutyric acid (1 mmol), triethylamine (1 mmol), 3 mL of N,N-dimethylformamide, and 3 mL of water. The reaction progress was monitored by TLC. After the reaction was complete, 15 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, washed once with saturated brine, dried over anhydrous sodium sulfate, the solvent was removed by distillation under reduced pressure, and the product was recrystallized from dichloromethane and ethyl ether. The solid was collected by filtration to obtain the pure product as a white solid in 96% yield.

[0133] Nuclear magnetic resonance and mass spectrometry experimental data of the product: 1 H NMR(400MHz,DMSO-d6)δ7.96-7.78(m,1H),7.23(td,J=17.3,15.8,6.9Hz,5H),6.81(d,J=8.3Hz,1H),4.16-3.99(m,1H),3.07(dq ,J=11.8,6.2Hz,2H),2.92(dd,J=13.6,4.8Hz,1H),2.81-2.67(m,1H),2.17(t,J=7.4Hz,2H),1.59(p,J=6.6Hz,2H),1.31(s,9H). 13C NMR(101MHz,DMSO)δ174.14,171.41,155.09,138.08,129.12,127.94,126.11,77.95,55.76,37.88,37.70,30.91,28.10,24.47. HRMS(ESI) m / z calculation for C 18 H 26 N2NaO5[M+Na] + :373.1734,found:373.1738.

[0134] Example 26 In this example, carfilzomib was produced, and the specific process is as follows:

[0135] [ka]

[0136] Chloroacetic acid (5 mmol) and N-methyl-N-toluenesulfonylethinamine (5.5 mmol) were dissolved in 5 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, L-homophenylalanine (10 mmol), triethylamine (10 mmol), acetonitrile (10 mL), and water (5 mL) were added to the previously stirred mixture and the reaction progress was monitored by TLC. After the reaction was completed, 20 mL of water was added, the mixture was acidified to pH 2 with 1 M hydrochloric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, washed once with saturated brine, and dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and ethyl ether, and the solid was collected by filtration to obtain the pure product. The above steps were repeated to sequentially condense L-phenylalanine, L-leucine, and (2S)-2-amino-4-methyl-1-[(2R)-2-methyloxiranyl]-1-pentanone trifluoroacetic acid. Finally, the obtained intermediate was reacted in a reaction bottle with morpholine (10 mmol), potassium iodide (KI, 5 mmol), and tetrahydrofuran (THF, 30 mL) under nitrogen gas protection at room temperature with stirring, and the reaction was monitored by TLC and HPLC. After the reaction was completed, the reaction mixture was concentrated, water and ethyl acetate were added, and the mixture was extracted three times with ethyl acetate. The organic layers were combined and dried over anhydrous magnesium sulfate. The reaction mixture was concentrated and recrystallized with ethyl acetate and petroleum ether to obtain carfilzomib, a white solid, with a total yield of 70%. The structural formula, nuclear magnetic resonance experimental data, and mass spectrometry experimental data of the product are shown below.

[0137] 1H NMR(400MHz,DMSO-d6)δ8.25(d,J=6.8Hz,1H),8.09(d,J=7.7Hz,1H),7.97(d,J=7.7Hz,1H),7.90(d,J=7.6Hz,1H), 7.33-7.23(m,2H),7.23-7.02(m,8H),4.61-4.49(m,1H),4.43-4.32(m,2H),4.32-4.23(m,1H),3.71-3.51(m,4H),3 .41-3.32(m,1H),3.12(d,J=4.2Hz,1H),3.06-2.85(m,4H),2.81-2.70(m,1H),2.57-2.51(m,1H),2.47-2.38(m,4H) ,1.93-1.76(m,2H),1.69-1.59(m,1H),1.55-1.47(m,1H),1.43-1.28(m,7H),0.89-0.83(m,6H),0.83-0.76(m,6H). 13 C NMR(100MHz,DMSO-d6)δ208.2,171.5,171.0,170.8,168.8,141.4,137.3,129.1,128.2,128.2,127.8,126.1,125.8,66. 1,61.3,58.7,53.2,53.0,51.8,51.5,51.1,49.2,40.7,38.5,37.4,34.3,31.4,24.5,24.1,23.2,23.0,21.6,21.0,16.3. HRMS(ESI) m / z calculation for C 40 H 58 N5O7[M+H] + :720.4331,found:720.4335.

[0138] Example 27 In this example, octreotide is prepared, and the specific process is as follows:

[0139] [ka]

[0140] Boc-D-Phe-OH (5 mmol) and N-methyl-N-toluenesulfonylethinamine (5.5 mmol) were dissolved in 20 mL of dichloromethane and stirred at room temperature. The reaction progress was monitored by TLC. After the starting acid was completely consumed, the solvent was removed by distillation under reduced pressure. H-Cys(Trt)-OH (10 mmol), N,N-diisopropylethylamine (10 mmol), acetonitrile (40 mL), and water (20 mL) were added to the previously stirred mixture and the reaction progress was monitored by TLC. After the reaction was completed, 50 mL of water was added, the mixture was acidified to pH 3 with 10% citric acid, the aqueous phase was extracted twice with ethyl acetate, the organic phases were combined, washed once with water, and washed once with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, and the mixture was recrystallized with dichloromethane and ethyl ether. The solid was collected by filtration to obtain Boc-D-Phe-Cys(Trt)-OH. The above steps were repeated to sequentially condense HD-Trp-OH, H-Lys(Boc)-OH, H-Thr(tBu)-OH, H-Cys(Trt)-OH, and L-Threoninol to obtain the protected linear octreotide, Boc-D-Phe-Cys(Trt)-Phe-D-Trp-Lys(Boc)-Thr(tBu)-Cys(Trt)-Thr-ol. This intermediate was added to a reaction bottle premixed with trifluoroacetic acid (42.5 mL), 1,2-ethanedithiol (2.5 mL), triisopropylsilane (2.5 mL), and water (2.5 mL), and the mixture was stirred. The reaction was monitored by TLC and HPLC. After completion of the reaction, the linear octreotide was recrystallized from ethyl ether to obtain the linear octreotide. Finally, the linear octreotide was dissolved in water to a concentration of 10. -4 After dilution to 100 mL, adjusting the pH to 8 with 1 mmol of aqueous ammonia, and oxidizing with air at room temperature for 48 h, octreotide was obtained as a white solid with an overall yield of 45%.

[0141] The structural formula, nuclear magnetic resonance experimental data, and mass spectrometry experimental data of the product are shown below.

[0142] 1H NMR(400MHz,DMSO-d6)δ10.88(s,1H),9.04(d,J=8.2Hz,1H),8.70(d,J=4.5Hz,1H),8.52(d,J=7.5Hz,1H),8.40(d,J=7.3Hz,1H),8.34(d,J=8.4Hz,1H),8.28-8.09(m,3H),7.98-7.85(m,3H),7.71(d,J=8.5Hz,1H),7.45(d,J=7.8Hz,1H),7.39-7.26(m,8H),7.21-7.18(m,2H),7.14-7.05(m,4H),7.02-6.97(m,2H),5.09(d,J=4.0Hz,1H),5.03-4.97(m,1H),4.90-4.85(m,1H),4.83-4.72(m,1H),4.64(s,1H),4.42(dd,J=8.4,5.4Hz,1H),4.24(dd,J=14.5,7.5Hz,2H),4.12-4.06(m,1H),4.00-3.92(m,2H),3.66-3.61(m,1H),3.18(dd,J=13.8,5.2Hz,1H),3.02-2.95(m,3H),2.92-2.72(m,7H),2.65-2.57(m,2H),1.73-1.64(m,1H),1.42-1.31(m,3H),1.11(d,J=6.2Hz,3H),1.05(d,J=6.4Hz,3H),0.89-0.77(m,2H). 13 C NMR(101MHz,DMSO-d6)δ172.53,171.87,170.65,170.39,169.59,168.47,168.10,136.81,136.14,134.76,129.66,129.04,128.54,128.09,127.23,127.05,126.44,123.69,120.90,118.26,118.18,111.34,109.03,66.63,64.23,60.20,58.58,55.90,55.22,53.97,53.36,53.23,52.55,52.01,44.17,42.39,38.78,38.52,37.42,30.26,26.44,26.38,22.03,19.92,19.50. HRMS(ESI)m / z calcd. for C 49 H 67N 10 O 10 S2 [M+H] + :1019.4478,found:1019.4483.

[0143] The present invention provides a method for synthesizing amides and / or polypeptides using unprotected amino acids as amino components, and in the method for producing polypeptides, after the construction of a peptide bond, the condensation of the next amino acid can be carried out directly without a deprotection operation, thereby reducing the use of many unnecessary protecting agents, deprotecting agents, and purification solvents, reducing the use of chemicals and the generation of chemical waste, significantly reducing the cost of polypeptide synthesis, improving the atom economy and step economy of polypeptide synthesis, greatly improving the efficiency of polypeptide synthesis, and effectively realizing energy savings and reducing pollutant emissions, and has the potential for broad industrial application.

[0144] Although the embodiments of the present invention have been described in detail above, the present invention is not limited to the above embodiments, and various modifications may be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features of the embodiments may be combined with each other unless inconsistent.

Claims

1. A method for producing an amide and / or a polypeptide, comprising the steps of: 【Chemical 1】 reacting a compound of formula IV with a compound of formula V in a solvent II, and then subjecting a compound of formula II and an alkali additive to a nucleophilic substitution reaction in a solvent I to prepare a compound of formula III; Here, R 1 is selected from an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue lacking a C-terminal carboxyl group, an amino acid derivative residue lacking a C-terminal carboxyl group, or a polypeptide fragment; R 2 is selected from hydrogen, deuterium, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a silyl group, or an alkylsilyl group; R 3 is selected from an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, and a heteroaryl group; R 4 is selected from an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue, or a polypeptide fragment; EWG is selected from a nitro group, a cyano group, a sulfonyl group, an arylsulfonyl group, an alkanoyl group, or a phosphonoyl group, wherein the alkyl group, alkenyl group, alkynyl group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, silyl group, or alkylsilyl group is optionally unsubstituted or is substituted with one or more R 11 and each R 11 are independently a halogen, a hydroxy group, a carbonyl group, C 1 ~C 6 A method for producing an amide and / or a polypeptide, wherein the alkyl group is selected from alkyl groups.

2. 2. The method for producing an amide and / or a polypeptide according to claim 1, wherein, when solvent II is different from solvent I, the method for producing an amide and / or a polypeptide further comprises a step of removing solvent II after the reaction and then performing the nucleophilic substitution reaction.

3. A method for producing an amide and / or a polypeptide, comprising the steps of: 【Chemistry 2】 The method comprises the steps of dissolving a compound of formula I, a compound of formula II and an alkaline additive in a solvent I, and carrying out a nucleophilic substitution reaction to produce a compound of formula III; Here, R 1 is selected from an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue lacking a C-terminal carboxyl group, an amino acid derivative residue lacking a C-terminal carboxyl group, or a polypeptide fragment; R 2 is selected from hydrogen, deuterium, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a silyl group, or an alkylsilyl group; R 3 is selected from an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, and a heteroaryl group; R 4 is selected from an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an amino acid residue, or a polypeptide fragment; EWG is selected from a nitro group, a cyano group, a sulfonyl group, an arylsulfonyl group, an alkanoyl group, or a phosphonoyl group, wherein the alkyl group, alkenyl group, alkynyl group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, silyl group, or alkylsilyl group is optionally unsubstituted or is substituted with one or more R 11 and each R 11 are independently a halogen, a hydroxy group, a carbonyl group, C 1 ~C 6 A method for producing an amide and / or a polypeptide, wherein the alkyl group is selected from alkyl groups.

4. The method for producing an amide and / or a polypeptide according to claim 1 or 3, wherein the alkaline additive comprises an organic base and / or an inorganic base.

5. Examples of the organic base include trimethylamine, triethylamine, tripropylamine, N,N-dimethylethylamine, N,N-diethylmethylamine, N,N-diisopropylethylamine, N-methylpyrrolidine, N-methylpiperidine, N-methylmorpholine, tributylamine, tribenzylamine, dimethylbenzylamine, N,N-dimethylaniline, pyridine, 2-methylpyridine, 2,6-dimethylpyridine, 4-dimethylaminopyridine, quinoline, quinine, tetramethylguanidine, imidazole, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylenediamine, tetramethylethylenediamine, and tetraethylethylenediamine.

5. The method for producing an amide and / or a polypeptide according to claim 4, wherein the inorganic base comprises at least one of an amine, tetramethylpropylenediamine, tetraethylpropylenediamine, tetrabutylammonium iodide, tetrabutylammonium fluoride, tetrabutylammonium chloride, tetrabutylammonium bromide, and tetrabutylammonium hydroxide, and the inorganic base comprises at least one of sodium bicarbonate, sodium carbonate, lithium bicarbonate, lithium carbonate, potassium bicarbonate, potassium carbonate, cesium bicarbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium bicarbonate, and calcium hydroxide.

6. The method for producing an amide and / or polypeptide according to claim 1 or 3, wherein the molar ratio of the alkaline additive to the compound of formula II is (0.1-20):

1.

7. 4. The method for producing an amide and / or a polypeptide according to claim 1 or 3, wherein the solvent I comprises at least one of water, acetonitrile, ethanol, isopropyl alcohol, tert-butanol, ethylene glycol, 1,2-propanediol, 2,3-butanediol, ethyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, toluene, m-xylene, dichloromethane, 1,2-dichloroethane, and chloroform, and the solvent II comprises at least one of dichloromethane, chloroform, 1,2-dichloroethane, ethyl ether, toluene, acetonitrile, methanol, and ethanol.

8. 4. The method for producing an amide and / or a polypeptide according to claim 1, wherein the temperature of the nucleophilic substitution reaction is -20°C to 150°C.

9. The method for producing an amide and / or polypeptide according to claim 1 or 3, characterized in that the compound of formula II includes any one of an amino acid compound, a polypeptide compound in which both the amino group and the carboxyl group are free, or other compounds having both an amino group and a carboxyl group.

10. The method for producing an amide and / or a polypeptide according to claim 3, comprising the steps of dissolving a compound of formula II and an alkaline additive in a solvent I, adding a compound of formula I, and carrying out a nucleophilic substitution reaction to produce a compound of formula III.

11. The method for preparing the compound of formula I comprises the steps of: 【Chemistry 3】 dissolving a compound of formula IV and a compound of formula V in a solvent II, and stirring and reacting them to produce a compound of formula I; Here, R 1 , R 2 , R 3 4. The method for producing an amide and / or polypeptide according to claim 3, wherein the definitions of EWG and EWG are as defined in claim 3.

12. The method for producing the amide and / or polypeptide includes the steps of: 【Chemistry 4】 Step S1: dissolving the compound of formula IV and the compound of formula V in a solvent II, stirring and reacting them to prepare a compound of formula I; Step S2 of dissolving the compound of formula I, the compound of formula II and an alkali additive in a solvent I and carrying out a nucleophilic substitution reaction to prepare a compound of formula III; Here, R 1 , R 2 , R 3 , R 4 4. The method for producing an amide and / or polypeptide according to claim 1 or 3, wherein the definitions of EWG are as defined in claim 1 or 3.

13. 13. The method for producing an amide and / or a polypeptide according to claim 12, wherein the compound of formula IV comprises at least one of a fatty acid, an aromatic acid, a heterocyclic acid, an alkynoic acid, an olefinic acid, an amino acid, a polypeptide carboxylic acid, and derivatives of the above compounds.

14. The method for producing an amide and / or polypeptide according to claim 12, wherein the molar ratio of the compound of formula IV to the compound of formula V is 1:(0.1-5).

15. Use of the method for producing an amide and / or a polypeptide according to any one of claims 1 to 14 in the production of an amide and / or a polypeptide.

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