Method for Synthesizing Amide and/or Polypeptide Using Temporary Protected Amino Acid as Ammonia Component

Abstract

The present disclosure discloses a method for synthesizing an amide and/or a polypeptide using a temporary protected amino acid as an ammonia component. In the present disclosure, a ynamide compound is used as a condensing agent, a low-cost unprotected amino acid is used as the raw material, a silylation reagent is used to temporarily protect the amino acid to perform a condensing reaction, and after the construction of a peptide bond is achieved, a target polypeptide carboxylic acid can be obtained by a simple acid treatment, which can be directly used for the condensation of the next amino acid.

Claims

1. A method for preparing an amide and/or a polypeptide, comprising the following steps: ##STR00033## reacting a compound of formula I with a compound of formula II in a solvent I, and adding a compound of formula III, a silylation reagent and a solvent II to perform a nucleophilic substitution reaction, and then obtaining a compound of formula IV after acidification; wherein R.sup.1 is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, an amino acid residual body lacking C-terminal carboxyl, and an amino acid derivative residual body or a polypeptide fragment lacking C-terminal carboxyl; R.sup.2 is selected from hydrogen, deuterium, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, a silyl or an alkylsilyl; R.sup.3 is selected from alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl; R.sup.4 is selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, an amino acid residue or a polypeptide fragment; EWG is selected from nitro, cyano, sulfonyl, arylsulfonyl, alkanoyl and phosphonyl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, silyl and alkylsilyl are optionally unsubstituted, or substituted by one or more R.sup.11; and each R.sup.11 is independently selected from halogen, hydroxyl, carbonyl and C.sub.1-C.sub.6 alkyl.

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

3. A method for preparing an amide and/or a polypeptide, comprising the following steps: ##STR00034## dissolving a compound of formula V, a compound of formula III and a silylation reagent in a solvent II to perform a nucleophilic substitution reaction, and then preparing a compound of formula IV after acidification; wherein R.sup.1 is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, an amino acid residual body lacking C-terminal carboxyl, and an amino acid derivative residual body or a polypeptide fragment lacking C-terminal carboxyl; R.sup.2 is selected from hydrogen, deuterium, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, a silyl or an alkylsilyl; R.sup.3 is selected from alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl; R.sup.4 is selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, an amino acid residue or a polypeptide fragment; EWG is selected from nitro, cyano, sulfonyl, arylsulfonyl, alkanoyl and phosphonyl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, silyl and alkylsilyl are optionally unsubstituted, or substituted by one or more R.sup.11; and each R.sup.11 is independently selected from halogen, hydroxyl, carbonyl and C.sub.1-C.sub.6 alkyl.

4. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the silylation reagent comprises at least one of N-trimethylsilylacetamide, N-methyl-N-trimethylsilylacetamide, N-trimethylsilylpyrrolidone, N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide, N,O-bis(tert-butyldimethylsilyl) acetamide, N,O-bistrimethylsilylacetamide, N, O-bis(trimethylsilyl)trifluoroacetamide, N-methyl-N-(trimethylsilyl)trifluoroacetamide, hexamethyldisilylurea, trimethylsilimidazole, dimethyl dichlorosilane, trimethylchlorosilane and tert-butyldimethylchlorosilane.

5. The method for preparing an amide and/or a polypeptide according to claim 3, wherein a molar ratio of the silylation reagent to the compound of formula III is (0.5-20):1.

6. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the solvent I comprises at least one of dichloromethane, chloroform, 1,2-dichloroethane, diethyl ether, toluene, acetonitrile, methanol or ethanol, and preferably the solvent II comprises at least one of acetonitrile, diethyl 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.

7. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the temperature of the nucleophilic substitution reaction is 20 C. to 150 C.

8. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the compound of formula III comprises any one of an amino acid compound, a polypeptide compound with free amino and carboxyl or other compounds with both amino and carboxyl.

9. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the method for preparing an amide and/or a polypeptide comprises the following steps: dissolving the compound of formula III and the silylation reagent in a solvent II, adding the compound of formula V to perform the nucleophilic substitution reaction, and then preparing the compound of formula IV after acidification.

10. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the method for preparing the compound of formula V comprises the following steps: ##STR00035## dissolving a compound of formula I and a compound of formula II in a solvent I to obtain a mixture, and reacting the mixture under stirring to prepare a compound of formula V; wherein R.sup.1, R.sup.2, R.sup.3 and EWG are defined as in claim 3.

11. The method for preparing an amide and/or a polypeptide according to claim 3, wherein the method for preparing an amide and/or a polypeptide comprises the following steps: ##STR00036## S1: dissolving the compound of formula I and the compound of formula II in the solvent I to obtain a mixture, and reacting the mixture under stirring to prepare a compound of formula V; and S2: dissolving the compound of formula V, the compound of formula III and the silylation reagent in the solvent II to perform the nucleophilic substitution reaction, and then preparing a compound of formula IV after acidification; wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and EWG are defined as in claim 3.

12. The method for preparing an amide and/or a polypeptide according to claim 11, wherein the compound of formula I comprises at least one of fatty acid, aromatic acid, heterocyclic acid, acetylenic acid, olefine acid, amino acid, polypeptide carboxylic acid and derivatives of the above compounds.

13. The method for preparing an amide and/or a polypeptide according to claim 11, wherein a molar ratio of the compound of formula I to the compound of formula III is 1:(0.1-5).

14. (canceled)

15. The method for preparing an amide and/or a polypeptide according to claim 1, wherein the silylation reagent comprises at least one of N-trimethylsilylacetamide, N-methyl-N-trimethylsilylacetamide, N-trimethylsilylpyrrolidone, N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide, N,O-bis(tert-butyldimethylsilyl) acetamide, N,O-bistrimethylsilylacetamide, N, O-bis(trimethylsilyl)trifluoroacetamide, N-methyl-N-(trimethylsilyl)trifluoroacetamide, hexamethyldisilylurea, trimethylsilimidazole, dimethyl dichlorosilane, trimethylchlorosilane and tert-butyldimethylchlorosilane.

16. The method for preparing an amide and/or a polypeptide according to claim 1, wherein a molar ratio of the silylation reagent to the compound of formula III is (0.5-20):1.

17. The method for preparing an amide and/or a polypeptide according to claim 1, wherein the solvent I comprises at least one of dichloromethane, chloroform, 1,2-dichloroethane, diethyl ether, toluene, acetonitrile, methanol or ethanol, and preferably the solvent II comprises at least one of acetonitrile, diethyl 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.

18. The method for preparing an amide and/or a polypeptide according to claim 1, wherein the temperature of the nucleophilic substitution reaction is 20 C. to 150 C.

19. The method for preparing an amide and/or a polypeptide according to claim 1, wherein the compound of formula III comprises any one of an amino acid compound, a polypeptide compound with free amino and carboxyl or other compounds with both amino and carboxyl.

20. The method for preparing an amide and/or a polypeptide according to claim 1, wherein the method for preparing an amide and/or a polypeptide comprises the following steps: ##STR00037## S1: dissolving the compound of formula I and the compound of formula II in the solvent I to obtain a mixture, and reacting the mixture under stirring to prepare a compound of formula V; and S2: dissolving the compound of formula V, the compound of formula III and the silylation reagent in the solvent II to perform the nucleophilic substitution reaction, and then preparing a compound of formula IV after acidification; wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and EWG are defined as in claim 1.

21. The method for preparing an amide and/or a polypeptide according to claim 19, wherein the compound of formula I comprises at least one of fatty acid, aromatic acid, heterocyclic acid, acetylenic acid, olefine acid, amino acid, polypeptide carboxylic acid and derivatives of the above compounds; wherein a molar ratio of the compound of formula I to the compound of formula III is 1:(0.1-5).

Description

DETAILED DESCRIPTION

[0043] The content of the present disclosure will be further described in detail below through specific examples. Unless otherwise specified, the raw materials, reagents or devices used in the examples and comparative examples can be obtained from conventional commercial sources, or can be obtained through existing technical methods. Unless otherwise stated, assays or testing methods are routine in the art.

Example 1

[0044] A polypeptide of Formula 1 was prepared in the example. A specific process was as follows:

##STR00010##

[0045] Boc-Val-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by Thin Layer Chromatography (TLC). When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Phe-OH (1 mmol), N-trimethylsilylacetamide (1 mmol) and 3 mL of N,N-dimethylformamide in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=2 with 10% citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 94%.

[0046] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0047] .sup.1H NMR (400 MHZ, DMSO) 8.04 (d, J=7.8 Hz, 1H), 7.30-7.16 (m, 5H), 6.54 (d, J=9.2 Hz, 1H), 4.47 (dd, J=13.3, 8.3 Hz, 1H), 3.78 (t, J=8.0 Hz, 1H), 3.06 (dd, J=13.9, 5.0 Hz, 1H), 2.90 (dd, J=13.9, 9.1 Hz, 1H), 1.97-1.77 (m, 1H), 1.38 (s, 9H), 0.83-0.71 (m, 6H).

[0048] .sup.13C NMR (100 MHz, DMSO) =172.8, 171.3, 155.2, 137.4, 129.1, 128.1, 126.4, 78.0, 59.7, 53.2, 36.8, 30.5, 28.2, 19.1, 18.1.

[0049] HRMS (ESI) m/z calcd. for C.sub.19H.sub.29N.sub.2O.sub.5 [M+H].sup.+: 365.2071, found: 365.2077.

Example 2

[0050] A polypeptide of Formula 2 was prepared in the example. A specific process was as follows:

##STR00011##

[0051] Cbz-Pro-OH (0.5 mmol), N-methyl-N-methylsulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Phe-OH (1 mmol), N,O-bistrimethylsilylacetamide (1 mmol) and 3 mL of dichloroethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=2 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure and subjected to column chromatography isolation to obtain a pure product, with a white solid, dr>99:1, and a yield of 93%.

[0052] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0053] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.38-8.03 (m, 1H), 7.42-7.11 (m, 10H), 5.12-5.02 (m, 1H), 5.01-4.82 (m, 1H), 4.53-4.40 (m, 1H), 4.23 (d, J=8.1 Hz, 1H), 3.40-3.29 (m, 2H), 3.12-3.00 (m, 1H), 2.99-2.84 (m, 2H), 2.13-1.96 (m, 1H), 1.80-1.59 (m, 3H).

[0054] .sup.13C NMR (100 MHZ, DMSO) =172.9, 172.8, 172.1, 171.8, 154.2, 153.8, 137.6, 137.5, 137.1, 129.2, 129.0, 128.4, 128.2, 128.1, 128.1, 127.8, 127.5, 127.0, 126.4, 126.3, 65.9, 65.7, 59.7, 59.3, 53.5, 53.1, 47.1, 46.5, 36.6, 31.0, 29.7, 23.7, 22.8.

[0055] HRMS (ESI) m/z calcd. for C.sub.22H.sub.25N.sub.2O.sub.5 [M+H].sup.+: 397.1758, found: 397.1764.

Example 3

[0056] A polypeptide of Formula 3 was prepared in the example. A specific process was as follows:

##STR00012##

[0057] Cbz-Phe-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.5 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Phe-OH (1 mmol), N,O-bistrimethylsilylacetamide (1 mmol) and 3 mL of N,N-dimethylformamide in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=2 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure and subjected to column chromatography isolation to obtain a pure product, with a white solid, dr>99:1, and a yield of 95%.

[0058] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0059] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.28 (d, J=7.7 Hz, 1H), 7.46 (d, J=8.7 Hz, 1H), 7.37-7.17 (m, 15H), 4.95 (s, 2H), 4.52 (dd, J=13.1, 7.7 Hz, 1H), 4.37-4.28 (m, 1H), 3.12 (dd, J=13.8, 5.0 Hz, 1H), 3.03-2.90 (m, 2H), 2.78-2.67 (m, 1H).

[0060] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 173.2, 172.0, 156.2, 138.5, 137.8, 137.5, 129.7, 128.7, 128.7, 128.5, 128.1, 127.9, 126.9, 126.7, 65.7, 56.5, 53.9, 37.9, 37.2.

[0061] HRMS (ESI) m/z calcd. for C.sub.26H.sub.27N.sub.2O.sub.6 [M+H].sup.+: 447.1914, found: 447.1919.

Example 4

[0062] A polypeptide of Formula 4 was prepared in the example. A specific process was as follows:

##STR00013##

[0063] Cbz-Thr-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Phe-OH (1 mmol), hexamethyldisilylurea (1 mmol) and 3 mL of acetonitrile in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=2 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 95%.

[0064] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0065] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.06 (d, J=7.7 Hz, 1H), 7.41-7.17 (m, 10H), 6.95 (d, J=8.7 Hz, 1H), 5.05 (s, 2H), 4.48 (q, J=7.3 Hz, 1H), 3.96 (dd, J=8.2, 5.2 Hz, 1H), 3.88-3.76 (m, 1H), 3.05 (dd, J=13.7, 5.0 Hz, 1H), 2.93 (dd, J=13.6, 8.1 Hz, 1H), 1.01 (d, J=6.1 Hz, 3H).

[0066] .sup.13C NMR (100 MHz, DMSO-d.sub.6) 172.6, 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.

[0067] HRMS (ESI) m/z calcd. for C.sub.21H.sub.24N.sub.2NaO.sub.6 [M+Na].sup.+: 423.1527, found: 423.1524.

Example 5

[0068] A polypeptide of Formula 5 was prepared in the example. A specific process was as follows:

##STR00014##

[0069] Boc-Met-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.5 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Phe-OH (1 mmol), N-Trimethylsilylimidazole (1 mmol) and 3 mL of acetonitrile in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure and subjected to column chromatography isolation to obtain a pure product, with a white solid, dr>99:1, and a yield of 97%.

[0070] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0071] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 7.97 (d, J=7.8 Hz, 1H), 7.34-7.16 (m, 5H), 6.91 (d, J=8.2 Hz, 1H), 4.46 (q, J=8.0 Hz, 1H), 4.01 (q, J=7.9 Hz, 1H), 3.07 (dd, J=13.8, 4.9 Hz, 1H), 2.92 (dd, J=13.9, 8.7 Hz, 1H), 2.45-2.29 (m, 2H), 2.01 (s, 3H), 1.85-1.64 (m, 2H), 1.37 (s, 9H).

[0072] .sup.13C NMR (100 MHz, DMSO-d.sub.6) 172.8, 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.

[0073] HRMS (ESI) m/z calcd. for C.sub.19H.sub.28N.sub.2NaO.sub.5S [M+Na].sup.+: 419.1611, found: 419.1609.

Example 6

[0074] A polypeptide of Formula 6 was prepared in the example. A specific process was as follows:

##STR00015##

[0075] Boc-Glu (O'Bu)-OH (0.5 mmol), N-methyl-N-methylsulfonyl ethynylamine (0.5 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Phe-OH (1 mmol), N, O-bis(trimethylsilyl)trifluoroacetamide (1 mmol) and 3 mL of dimethyl sulfoxide in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure and subjected to column chromatography isolation to obtain a pure product, with a white solid, dr>99:1, and a yield of 94%.

[0076] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0077] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 7.95 (d, J=7.9 Hz, 1H), 7.36-7.10 (m, 5H), 6.82 (d, J=8.3 Hz, 1H), 4.52-4.40 (m, 1H), 4.01-3.85 (m, 1H), 3.07 (dd, J=13.8, 4.8 Hz, 1H), 2.92 (dd, J=13.9, 8.8 Hz, 1H), 2.16 (t, J=7.6 Hz, 2H), 1.85-1.72 (m, 1H), 1.70-1.58 (m, 1H), 1.40 (s, 9H), 1.37 (s, 9H).

[0078] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 172.7, 171.7, 171.5, 155.1, 137.3, 129.2, 128.1, 126.4, 79.5, 78.2, 53.5, 53.2, 36.7, 31.2, 28.1, 27.8, 27.4.

[0079] HRMS (ESI) m/z calcd. for C.sub.23H.sub.34N.sub.2O.sub.7 [M+H].sup.+: 451.2439, found: 451.2445.

Example 7

[0080] A polypeptide of Formula 7 was prepared in the example. A specific process was as follows:

##STR00016##

[0081] Cbz-Ala-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of trichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Thr-OH (1 mmol), N,O-bistrimethylsilylacetamide (2 mmol) and 3 mL of trichloromethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=2 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with ethyl acetate and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 92%.

[0082] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0083] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 7.67 (d, J=8.6 Hz, 1H), 7.53 (d, J=7.7 Hz, 1H), 7.39-7.26 (m, 5H), 5.03 (s, 2H), 4.25-4.14 (m, 3H), 1.24 (d, J=7.1 Hz, 3H), 1.06 (d, J=6.1 Hz, 3H).

[0084] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 173.0, 172.2, 155.8, 137.1, 128.5, 127.9, 127.8, 66.5, 65.5, 57.5, 50.2, 20.4, 18.2.

[0085] HRMS (ESI) m/z calcd. for C.sub.15H.sub.20N.sub.2O.sub.6 [M+Na].sup.+: 347.1214, found: 347.1221.

Example 8

[0086] A polypeptide of Formula 8 was prepared in the example. A specific process was as follows:

##STR00017##

[0087] Cbz-Ala-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Gln-OH (1 mmol), N,O-bistrimethylsilylacetamide (2 mmol) and 3 mL of N,N-dimethylacetamide in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=1 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 95%.

[0088] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0089] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.13 (d, J=7.6 Hz, 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=7.1 Hz, 3H).

[0090] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 173.6, 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.

[0091] HRMS (ESI) m/z calcd. for C.sub.16H.sub.21N.sub.3NaO.sub.6 [M+Na].sup.+: 374.1323, found: 374.1321.

Example 9

[0092] A polypeptide of Formula 9 was prepared in the example. A specific process was as follows:

##STR00018##

[0093] Cbz-Ala-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Trp-OH (1 mmol), hexamethyldisilylurea (1 mmol) and 3 mL of N-methylpyrrolidone in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=1 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure and subjected to column chromatography isolation to obtain a pure product, with a white solid, dr>99:1, and a yield of 95%.

[0094] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0095] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 10.89 (s, 1H), 8.12-8.03 (m, 1H), 7.57 (t, J=6.0 Hz, 1H), 7.44 (dd, J=8.1, 4.1 Hz, 1H), 7.34 (d, J=15.7 Hz, 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, 4.9 Hz, 2H), 3.22 (d, J=14.3 Hz, 2H), 3.17-3.05 (m, 2H), 1.26-1.19 (m, 7H).

[0096] .sup.13C NMR (100 MHz, DMSO-d.sub.6) 173.3, 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.

[0097] HRMS (ESI) m/z calcd. for C.sub.22H.sub.23N.sub.3NaO.sub.5 [M+Na].sup.+: 432.1530, found: 432.1530.

Example 10

[0098] A polypeptide of Formula 10 was prepared in the example. A specific process was as follows:

##STR00019##

[0099] Fmoc-Ala-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 3 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Phe-OH (1 mmol), N-methyl-N-trimethylsilylacetamide (1 mmol) and 3 mL of dichloromethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=1 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure and subjected to column chromatography isolation to obtain a pure product, with a white solid, dr>99:1, and a yield of 93%.

[0100] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0101] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.02 (d, J=7.5 Hz, 1H), 7.89 (d, J=7.5 Hz, 2H), 7.74 (d, J=6.8 Hz, 1H), 7.48 (d, J=7.7 Hz, 1H), 7.42 (t, J=7.4 Hz, 2H), 7.33 (t, J=7.3 Hz, 2H), 7.27-7.16 (m, 4H), 4.50-4.39 (m, 1H), 4.26 (d, J=6.1 Hz, 1H), 4.24-4.18 (m, 1H), 4.12-4.06 (m, 1H), 3.07 (dd, J=13.7, 4.8 Hz, 1H), 2.93 (dd, J=13.7, 8.4 Hz, 1H), 1.20 (d, J=7.1 Hz, 3H).

[0102] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 172.8, 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.

[0103] HRMS (ESI) m/z calcd. for C.sub.22H.sub.23N.sub.3NaO.sub.5 [M+Na].sup.+: 432.1530, found: 432.1530.

Example 11

[0104] A polypeptide of Formula 11 is prepared in the example. A specific process is as follows:

##STR00020##

[0105] Boc-Phe-Ala-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.5 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Ser (O'Bu)-OH (1 mmol), N-trimethylsilylpyrrolidone (1 mmol) and 5 mL of 1,2-dichloroethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with dichloromethane, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 94%.

[0106] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0107] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.04 (d, J=7.4 Hz, 1H), 7.94 (d, J=7.9 Hz, 1H), 7.32-7.15 (m, 5H), 6.94 (d, J=8.5 Hz, 1H), 4.49-4.41 (m, 1H), 4.40-4.30 (m, 1H), 4.22-4.11 (m, 1H), 3.69-3.60 (m, 1H), 3.54-3.48 (m, 1H), 3.05-2.94 (m, 1H), 2.78-2.65 (m, 1H), 1.29 (s, 9H), 1.24 (d, J=6.8 Hz, 3H), 1.11 (s, 9H)

[0108] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 172.1, 171.4, 171.3, 155.2, 138.2, 129.2, 127.9, 126.1, 78.0, 72.8, 61.5, 55.6, 52.8, 47.8, 37.4, 28.1, 27.1, 18.4.

[0109] HRMS (ESI) m/z calcd. for C.sub.24H.sub.38N.sub.3O.sub.7 [M+H].sup.+: 480.2704, found: 480.2710.

Example 12

[0110] A polypeptide of Formula 12 was prepared in the example. A specific process was as follows:

##STR00021##

[0111] Cbz-Ala-Leu-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.5 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Phe-OH (1 mmol), N,O-bistrimethylsilylacetamide (1 mmol) and 5 mL of dichloromethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=1 with 2 M of hydrochloric acid. An aqueous phase was extracted twice with dichloromethane, an organic phase was combined, washed once with water, washed once with a saturated salt solution, and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 91%.

[0112] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0113] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.04 (d, J=7.6 Hz, 1H), 7.86 (d, J=8.3 Hz, 1H), 7.43 (d, J=7.6 Hz, 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, 5.2 Hz, 1H), 2.93 (dd, J=13.8, 8.5 Hz, 1H), 1.67-1.50 (m, 1H), 1.43 (d, J=6.7 Hz, 1H), 1.17 (d, J=7.0 Hz, 3H), 0.90-0.76 (m, 6H).

[0114] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 1172.8, 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.

[0115] HRMS (ESI) m/z calcd. For C.sub.26H.sub.34N.sub.3O.sub.6 [M+H].sup.+: 484.2442, found: 284.2448.

Example 13

[0116] A polypeptide of Formula 13 was prepared in the example. A specific process was as follows:

##STR00022##

[0117] Cbz-Ala-Ala-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Gln(Trt)-OH (1 mmol), N,O-bistrimethylsilylacetamide (2 mmol) and 5 mL of dichloromethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted once with ethyl acetate and washed twice with water, an organic phase was washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a light yellow solid, dr>99:1, and a yield of 95%.

[0118] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0119] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.59 (s, 1H), 8.05 (d, J=8.2 Hz, 1H), 7.93 (t, J=6.5 Hz, 1H), 7.44 (t, J=5.9 Hz, 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).

[0120] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 173.3, 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.

[0121] HRMS (ESI) m/z calcd. for C.sub.38H.sub.40N.sub.4NaO.sub.7 [M+Na].sup.+: 687.2789, found: 687.2874.

Example 14

[0122] A polypeptide of Formula 14 was prepared in the example. A specific process was as follows:

##STR00023##

[0123] Boc-Ala-Phe-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Tyr(O'Bu)-OH (1 mmol), N,O-bistrimethylsilylacetamide (2 mmol) and 5 mL of dichloromethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted once with ethyl acetate and washed twice with water, an organic phase was washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with diethyl ether and petroleum ether, and the solid was filtered and collected to obtain a pure product, with a light yellow solid, dr>99:1, and a yield of 91%.

[0124] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0125] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.35-8.22 (m, 1H), 7.80-7.66 (m, 1H), 7.27-7.11 (m, 7H), 6.94-6.78 (m, 3H), 4.64-4.52 (m, 1H), 4.48-4.37 (m, 1H), 3.99-3.82 (m, 1H), 3.07-2.96 (m, 2H), 2.93-2.84 (m, 1H), 2.82-2.69 (m, 1H), 1.36 (s, 9H), 1.26 (s, 9H), 1.06 (d, J=6.6 Hz, 3H).

[0126] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 172.64, 172.15, 170.76, 154.86, 153.59, 137.44, 131.81, 129.53, 129.28, 127.86, 126.12, 123.34, 78.09, 77.58, 53.41, 53.20, 49.81, 37.67, 36.04, 28.53, 28.14, 18.14.

[0127] HRMS (ESI) m/z calcd. for C.sub.30H.sub.41N.sub.3NaO.sub.7 [M+Na].sup.+: 578.2837, found: 578.2840.

Example 15

[0128] A polypeptide of Formula 15 was prepared in the example. A specific process was as follows:

##STR00024##

[0129] Cbz-Ala-Phe-Met-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Glu (O'Bu)-OH (1 mmol), hexamethyldisilylurea (1 mmol) and 3 mL of dichloromethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water and washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 92%.

[0130] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0131] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.13 (d, J=7.5 Hz, 1H), 8.07 (d, J=7.4 Hz, 1H), 7.93 (d, J=7.3 Hz, 1H), 7.43 (d, J=7.1 Hz, 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=7.0 Hz, 3H).

[0132] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 173.0, 172.4, 171.5, 170.9, 170.7, 155.7, 137.6, 136.9, 129.3, 128.3, 127.9, 127.8, 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.

[0133] HRMS (ESI) m/z calcd. for C.sub.34H.sub.47N.sub.4O.sub.9S [M+H].sup.+: 687.3058, found: 687.3065.

Example 16

[0134] A polypeptide of Formula 16 was prepared in the example. A specific process was as follows:

##STR00025##

[0135] Cbz-Phe-Gly-Ala-Glu (tBu)-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that mixed with H-Phe-OH (1 mmol), N-methyl-N-trimethylsilylacetamide (2 mmol) and 5 mL of acetonitrile in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 90%.

[0136] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0137] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.43-8.25 (m, 1H), 8.12-7.89 (m, 3H), 7.57 (d, J=7.1 Hz, 1H), 7.37-7.13 (m, 15H), 5.04-4.87 (m, 2H), 4.52-4.42 (m, 1H), 4.40-4.24 (m, 3H), 3.84-3.71 (m, 2H), 3.12-3.01 (m, 2H), 2.99-2.89 (m, 1H), 2.84-2.72 (m, 1H), 2.32-2.13 (m, 2H), 1.96-1.83 (m, 1H), 1.81-1.66 (m, 1H), 1.39 (s, 9H), 1.20 (d, J=6.2 Hz, 3H).

[0138] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 172.7, 171.9, 171.9, 171.8, 170.9, 168.4, 156.0, 138.1, 137.3, 137.0, 129.2, 129.1, 128.3, 128.2, 128.0, 127.7, 127.4, 126.4, 126.2, 79.6, 65.3, 56.3, 53.4, 51.6, 48.1, 42.1, 37.3, 36.6, 31.2, 27.8, 27.5, 18.2.

[0139] HRMS (ESI) m/z calcd. for C.sub.40H.sub.50N.sub.5O.sub.10 [M+H].sup.+: 760.3552, found: 760.3557.

Example 17

[0140] A polypeptide of Formula 17 was prepared in the example. A specific process was as follows:

##STR00026##

[0141] Boc--Ala-Trp-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that was mixed with H-Met-OH (1 mmol), N, O-bis(trimethylsilyl)trifluoroacetamide (1 mmol) and 3 mL of acetonitrile in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 94%.

[0142] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0143] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 10.80 (s, 1H), 8.28 (d, J=7.7 Hz, 1H), 8.06 (d, J=8.1 Hz, 1H), 7.62 (d, J=7.8 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.15 (s, 1H), 7.06 (t, J=7.4 Hz, 1H), 6.98 (t, J=7.4 Hz, 1H), 6.65 (t, J=5.2 Hz, 1H), 4.66-4.53 (m, 1H), 4.41-4.29 (m, 1H), 3.18-3.01 (m, 3H), 2.96-2.85 (m, 1H), 2.52-2.41 (m, 2H), 2.29-2.14 (m, 2H), 2.04 (s, 3H), 2.02-1.95 (m, 1H), 1.95-1.84 (m, 1H), 1.36 (s, 9H).

[0144] .sup.13C NMR (100 MHz, DMSO-d.sub.6) 173.2, 171.9, 170.3, 155.5, 136.1, 127.4, 123.6, 120.9, 118.5, 118.2, 111.3, 110.2, 77.6, 53.2, 51.0, 36.7, 35.7, 30.8, 29.7, 28.3, 27.7, 14.7.

[0145] HRMS (ESI) m/z calcd. for C.sub.24H.sub.35N.sub.4O.sub.6S [M+H].sup.+: 507.2272, found: 507.2278.

Example 18

[0146] A polypeptide of Formula 18 was prepared in the example. A specific process was as follows:

##STR00027##

[0147] Cbz-Tyr(tBu)-Gly-Gly-Phe-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the dichloromethane was removed by distillation under reduced pressure and transferred to a system that mixed with H-Leu-OH (1 mmol), N,O-bistrimethylsilylacetamide (1 mmol) and 5 mL of N,N-dimethylformamide in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 90%.

[0148] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0149] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.35-8.20 (m, 2H), 8.09-7.93 (m, 2H), 7.60-7.46 (m, 1H), 7.38-7.14 (m, 12H), 6.85 (d, J=7.9 Hz, 2H), 5.04-4.84 (m, 2H), 4.70-4.54 (m, 1H), 4.36-4.20 (m, 2H), 3.82-3.70 (m, 3H), 3.69-3.62 (m, 1H), 3.12-2.97 (m, 2H), 2.84-2.65 (m, 2H), 1.74-1.49 (m, 3H), 1.26 (s, 9H), 0.97-0.80 (m, 6H).

[0150] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 173.9, 171.9, 171.1, 168.9, 168.3, 155.9, 153.5, 137.8, 137.0, 132.7, 129.7, 129.3, 128.3, 128.0, 127.7, 127.5, 126.2, 123.3, 77.6, 65.3, 56.3, 53.6, 50.4, 42.1, 41.8, 40.0, 37.7, 36.8, 28.6, 24.3, 22.8, 21.4.

[0151] HRMS (ESI) m/z calcd. for C.sub.40H.sub.52N.sub.5O.sub.9 [M+H].sup.+: 746.3760, found: 746.3766.

Example 19

[0152] A polypeptide of Formula 19 was prepared in the example. A specific process was as follows:

##STR00028##

[0153] Cbz-Tyr(tBu)-Gly-Gly-OH (0.5 mmol), N-methyl-N-p-toluenesulfonyl ethynylamine (0.55 mmol) were taken to be dissolved in 5 mL of dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that mixed with H-Phe-Met-OH (1 mmol), N-trimethylsilylpyrrolidone (1 mmol) and 5 mL of dichloroethane in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 15 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product, with a white solid, dr>99:1, and a yield of 93%.

[0154] Magnetic resonance imaging experimental data and mass spectrometry experimental data of products were as follows:

[0155] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.40-8.20 (m, 2H), 8.16-8.07 (m, 1H), 8.07-7.97 (m, 1H), 7.54 (d, J=8.1 Hz, 1H), 7.35-7.15 (m, 12H), 6.86 (d, J=7.6 Hz, 2H), 4.94 (s, 2H), 4.64-4.52 (m, 1H), 4.38-4.24 (m, 2H), 3.79-3.63 (m, 4H), 3.11-2.97 (m, 2H), 2.86-2.69 (m, 2H), 2.51-2.41 (m, 2H), 2.08-1.96 (m, 4H), 1.94-1.86 (m, 1H), 1.26 (s, 9H).

[0156] .sup.13C NMR (100 MHz, DMSO) 173.3, 172.0, 171.1, 169.0, 168.5, 156.0, 153.5, 137.8, 137.0, 132.8, 129.7, 129.3, 128.3, 128.1, 127.7, 127.5, 126.3, 123.3, 77.6, 65.4, 56.3, 53.8, 51.3, 42.2, 41.9, 37.6, 36.8, 30.9, 29.7, 28.6, 14.6.

[0157] HRMS (ESI) m/z calcd. for C.sub.39H.sub.50N.sub.5O.sub.9S [M+H].sup.+: 764.3324, found: 764.3330.

Example 20

[0158] Carfilzomib was prepared in the example. A specific process was as follows:

##STR00029##

[0159] Chloroacetic acid (5 mmol) and N-methyl-N-p-toluenesulfonyl ethynylamine (5.5 mmol) were taken to be dissolved in 5 mL dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with L-homophenylalanine (10 mmol), N,O-bistrimethylsilylacetamide (10 mmol) and dichloromethane (5 mL) in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 20 mL of water was added to acidify to pH=2 with 1 M of hydrochloric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain a pure product. The above steps were repeated to successively condense with L-phenylalanine, L-leucine and (2S)-2-amino-4-methyl-1-[(2R)-2-methyloxiranyl]-1-pentanone trifluoroacetate, the finally obtained intermediate was in a room-temperature stirring reaction with morpholine (10 mmol), potassium iodide (KI, 5 mmol) and tetrahydrofuran (THF, 30 mL) in a reaction flask under nitrogen protection, and the reaction was monitored by TLC and High Performance Liquid Chromatography (HPLC). After the reaction was completed, the reaction solution was concentrated, water and ethyl acetate were added, the organic phase was extracted three times with ethyl acetate, combined and dried with anhydrous magnesium sulfate, the reaction solution was concentrated and recrystallized with ethyl acetate and petroleum ether to obtain the carfilzomib, with a white solid and a total yield of 65%. The following was the structural formula, magnetic resonance imaging experimental data and mass spectrometry experimental data of the product:

[0160] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.25 (d, J=6.8 Hz, 1H), 8.09 (d, J=7.7 Hz, 1H), 7.97 (d, J=7.7 Hz, 1H), 7.90 (d, J=7.6 Hz, 1H), 7.33-7.23 (m, 2H), 7.22-7.02 (m, 8H), 4.61-4.49 (m, 1H), 4.42-4.33 (m, 2H), 4.32-4.23 (m, 1H), 3.71-3.50 (m, 4H), 3.41-3.32 (m, 1H), 3.12 (d, J=4.2 Hz, 1H), 3.05-2.85 (m, 4H), 2.80-2.70 (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).

[0161] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 208.2, 171.5, 171.0, 170.9, 168.8, 141.4, 137.3, 129.1, 128.2, 128.2, 127.9, 126.1, 125.8, 66.0, 61.3, 58.7, 53.2, 53.0, 51.8, 51.5, 51.2, 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.

[0162] HRMS (ESI) m/z calcd. for C.sub.40H.sub.58N.sub.5O.sub.7 [M+H].sup.+: 720.4331, found: 720.4330.

Example 21

[0163] Octreotide was prepared in the example. A specific process was as follows:

##STR00030##

[0164] Boc-D-Phe-OH (5 mmol) and N-methyl-N-p-toluenesulfonyl ethynylamine (5.5 mmol) were taken to be dissolved in 20 mL dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Cys(Trt)-OH (10 mmol), N,O-bistrimethylsilylacetamide (10 mmol) and dichloromethane (40 mL) in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 50 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain Boc-D-Phe-Cys(Trt)-OH. The above steps were repeated to successively condense with H-D-Trp-OH, H-Lys(Boc)-OH, H-Thr (tBu)-OH, HCys(Trt)-OH and L-threoninol, to obtain the protected octreotide straight chain Boc-D-Phe-Cys(Trt)-Phe-D-Trp-Lys (Boc)-Thr (tBu)-Cys(Trt)-Thr-ol, the intermediate was added to a reaction flask that mixed with trifluoroacetic acid (42.5 mL), 1,2-ethanedithiol (2.5 mL), triisopropylsilane (2.5 mL) and water (2.5 mL) in advance, stirring was performed and the reaction was monitored by TLC and HPLC. After the reaction was completed, the reaction solution was concentrated, recrystallization was performed with diethyl ether to obtain the octreotide straight chain. Finally, the octreotide straight chain was dissolved in water, the concentration was diluted to 10.sup.4M, pH=8 was adjusted with 1 mmol of ammonium hydroxide, air was accessed at a room temperature for 48 h of oxidation, the octreotide was obtained after freeze-drying, with a white solid and a total yield of 56%. The following was the structural formula, magnetic resonance imaging experimental data and mass spectrometry experimental data of the product:

[0165] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 10.88 (s, 1H), 9.04 (d, J=8.2 Hz, 1H), 8.70 (d, J=4.5 Hz, 1H), 8.52 (d, J=7.5 Hz, 1H), 8.40 (d, J=7.3 Hz, 1H), 8.34 (d, J=8.4 Hz, 1H), 8.28-8.09 (m, 3H), 7.98-7.85 (m, 3H), 7.71 (d, J=8.5 Hz, 1H), 7.45 (d, J=7.8 Hz, 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=4.0 Hz, 1H), 5.03-4.97 (m, 1H), 4.90-4.84 (m, 1H), 4.83-4.72 (m, 1H), 4.64 (s, 1H), 4.42 (dd, J=8.4, 5.4 Hz, 1H), 4.24 (dd, J=14.5, 7.5 Hz, 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, 5.2 Hz, 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=6.2 Hz, 3H), 1.05 (d, J=6.4 Hz, 3H), 0.89-0.77 (m, 2H).

[0166] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 172.53, 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.

[0167] HRMS (ESI) m/z calcd. for C.sub.49H.sub.67N.sub.10O.sub.10S.sub.2 [M+H].sup.+: 1019.4478, found: 1019.4484.

Example 22

[0168] A protected lypressin straight chain was prepared in the example. A specific process was as follows:

##STR00031## ##STR00032##

[0169] Boc-Cys(Trt)-OH (5 mmol) and N-methyl-N-p-toluenesulfonyl ethynylamine (5.5 mmol) were taken to be dissolved in 20 mL dichloromethane, a stirring reaction was performed at a room temperature, and the reaction process was monitored by TLC. When the raw acid was completely consumed, the mixture was transferred to a system that was mixed with H-Tyr(O'Bu)-OH (10 mmol), N,O-bistrimethylsilylacetamide (10 mmol) and dichloromethane (40 mL) in advance for reaction, and the reaction process was monitored by TLC. After the reaction was completed, 50 mL of water was added to acidify to pH=3 with 10% of citric acid. An aqueous phase was extracted twice with ethyl acetate, an organic phase was combined, washed once with water, washed once with a saturated salt solution and dried with anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure, recrystallized with dichloromethane and diethyl ether, and the solid was filtered and collected to obtain Boc-Cys(Trt)-Tyr(O'Bu)-OH. The above steps were repeated to successively condense with H-Phe-OH, H-Gln(Trt)-OH, H-Asn(Trt)-OH, HCys(Trt)-OH, HPro-OH, and H-Lys(Boc)-OH, H-Gly-NH.sub.2 to obtain the protected lypressin straight chain Boc-Cys(Trt)-Tyr(O'Bu)-Phe-Gln(Trt)-Asn(Trt)-Cys(Trt)-Pro-Lys(Boc)-Gly-NH.sub.2, with a white solid and a total yield of 55%. The following was the structural formula, magnetic resonance imaging experimental data and mass spectrometry experimental data of the product:

[0170] .sup.1H NMR (400 MHZ, DMSO-d.sub.6) 8.67-8.60 (m, 1H), 8.46-7.94 (m, 8H), 7.65-7.54 (m, 2H), 7.42-7.31 (m, 24H), 7.24-6.96 (m, 43H), 6.81-6.75 (m, 1H), 6.73-6.46 (m, 3H), 4.76-4.64 (m, 2H), 4.53-4.43 (m, 2H), 4.30-4.20 (m, 2H), 4.15-4.07 (m, 1H), 3.87-3.75 (m, 1H), 3.69-3.59 (m, 2H), 3.52-3.46 (m, 2H), 3.17-3.05 (m, 2H), 2.98-2.80 (m, 5H), 2.77-2.65 (m, 3H), 2.62-2.53 (m, 2H), 2.46-2.33 (m, 4H), 2.29-2.09 (m, 2H), 2.04-1.89 (m, 2H), 1.84-1.73 (m, 2H), 1.66-1.57 (m, 2H), 1.44-1.37 (m, 18H), 1.34-1.28 (m, 4H), 1.20 (s, 9H).

[0171] .sup.13C NMR (100 MHZ, DMSO-d.sub.6) 171.81, 171.77, 171.57, 171.34, 171.15, 170.85, 170.76, 170.66, 169.27, 168.60, 168.18, 155.57, 154.83, 153.24, 144.92, 144.71, 144.31, 144.26, 137.67, 132.03, 129.86, 129.84, 129.19, 129.11, 129.07, 128.58, 128.55, 128.07, 127.99, 127.46, 127.44, 126.70, 126.31, 126.14, 123.24, 78.53, 77.39, 69.42, 69.26, 66.45, 65.85, 59.31, 53.74, 53.14, 52.93, 52.01, 50.90, 49.79, 46.42, 41.98, 37.43, 37.33, 33.93, 32.72, 32.46, 31.30, 31.24, 29.30, 29.24, 29.17, 28.56, 28.32, 28.15, 24.25, 22.69, 22.63.

[0172] HRMS (ESI) m/z calcd. for C.sub.136H.sub.148N.sub.13O.sub.16S.sub.2 [M+H].sup.+: 2283.0603, found: 2283.0610.

[0173] The present disclosure discloses a method for synthesizing an amide and/or a polypeptide using a temporary protected amino acid as an ammonia component. In the present disclosure, the ynamide compound is used as the condensation agent, a low-cost unprotected amino acid is used as the raw material, and the silylation reagent is used to temporary protect the amino acid to perform the condensation reaction, so that protection-condensation-deprotection are carried out in one pot, and the three steps of reaction are combined into one step. After the construction of the peptide bond is achieved, the target polypeptide carboxylic acid can be obtained by a simple acid treatment, which can be directly used for the condensation of the next amino acid. At the same time, based on the characteristics of ynamide condensation agent, this method can also completely avoid the formation of racemic by-products, to ensure the yield and purity of the product. This method saves a large number of reagents and solvents used in the traditional methods of protecting groups on amino acids and product deprotection, reduces the generation of a large amount of chemical waste, greatly reduces the cost of peptide synthesis, and improves the atom economy and step economy of peptide synthesis.

[0174] The above examples are preferred examples of the present disclosure, but the embodiments of the present disclosure are not limited to the above examples. Any other changes, modifications, substitutions, combinations and simplification made without departing from the spirit and principles of the present disclosure should be equivalent substitutions, and all included in the protection scope of the present disclosure.