a-CARBONYL ALKENYL ESTER PREPARATION METHOD THEREFOR AND APPLICATION THEREOF

Abstract

There is provided an α-carbonyl alkenyl ester and a preparation method therefor, and the α-carbonyl alkenyl ester is further used to react with a primary or secondary amine to prepare an amide. The two reactions are combined to develop an amide bond and peptide bond formation method that directly use carboxylic acids and amines as starting materials and allenones as a condensing reagent. The α-carbonyl alkenyl ester corresponding to an α-amino acid serves as a peptide synthesis building block and is used in solid phase peptide synthesis. The method is carried out under mild reaction conditions, simple to operate, and has a high yield. Compared with existing amide bond condensation reagents, the allenones have the advantages of being simple to prepare, having good stability, a low molecular weight, not racemizing when activating α-chiral carboxylic acids, and is a novel amide bond and peptide bond condensing reagent.

Claims

1. An α-carbonyl alkenyl ester having the general formula (I): ##STR00117## wherein R.sup.1 is selected from one of C1-C24 hydrocarbyl, C1-C24 hydrocarbyl substituted by a substituent(s), protected α-amino C1-C20 hydrocarbyl, protected β-amino C2-C20 hydrocarbyl, protected γ-amino C3-C20 hydrocarbyl, and protected polypeptide chain C1-C20 hydrocarbyl; R.sup.2 is selected from one of aryl, aryl substituted by a substituent(s), heteroaryl and heteroaryl substituted by a substituent(s); R.sup.3, R.sup.4 and R.sup.5 are the same or different, and are each independently selected from one of H, C1-C18 hydrocarbyl, C1-C18 hydrocarbyl substituted by a substituent(s), C1-C16 acyl, cyano, halogen or C1-C16 hydrocarbyloxy-carbonyl.

2. The compound as claimed in claim 1, wherein R.sup.1 is selected from one of protected α-amino C2-C10 hydrocarbyl, protected β-amino C3-C10 hydrocarbyl, protected γ-amino C4-C10 hydrocarbyl, and protected polypeptide chain C2-C15 hydrocarbyl; R.sup.2 is selected from one of C6-C24 aryl, C6-C24 aryl substituted by a substituent(s), C4-C24 heteroaryl, and C4-C24 heteroaryl substituted by a substituent(s); R.sup.3, R.sup.4 and R.sup.5 are the same or different, and are each independently selected from one of H C1-C12 hydrocarbyl, C1-C12 hydrocarbyl substituted by a substituent(s), C1-C16 acyl, cyano, halogen or C2-C12 hydrocarbyloxy-carbonyl.

3. The compound as claimed in claim 1, wherein, R.sup.1 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, adamantyl, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, phenylethynyl, phenyl, naphthyl, anthracenyl, phenanthryl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, pyrrolyl, indolyl, indolylmethyl, indazolyl, furyl, benzofuranyl, thienyl, benzothienyl, styryl, phenylethynyl, benzyl, 11-hydroxyundecyl, pentadecyl, protected α-amino C1-C20 alkyl, protected β-amino C2-C20 alkyl, protected γ-amino C3-C20 alkyl, and protected polypeptide chain C1-C20 alkyl; and/or R.sup.2 is selected from one of phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 2-methylphenyl, 2-methoxyphenyl, 3,5-dimethoxypropyl, 3-nitrophenyl, nitrophenyl, 2,4-dinitrophenyl, 3,5-dinitrophenyl, pentafluorophenyl, 4-trifluoromethylphenyl, 3,5-dichlorophenyl, 1-naphthyl, 2-naphthyl, furanyl, thienyl, and pyridyl; and/or R.sup.3, R.sup.4, R.sup.5 are each independently selected from one of H, methyl, formyl, acetyl, propionyl, butyryl, cyano, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl and benzyloxycarbonyl.

4. (canceled)

5. The compound as claimed in claim 1, wherein, the compound having the general formula (I) is one selected from the following compounds: (E)-4-oxo-4-phenylbut-2-en-2-yl acetate: ##STR00118## (E)-4-oxo-4-phenylbut-2-en-2-yl (3r,5r,7r)-adamantane-1-carboxylate: ##STR00119## (E)-4-oxo-4-phenylbut-2-en-2-yl 4-chlorobenzoate: ##STR00120## (E)-4-oxo-4-phenylbut-2-en-2-yl furan-2-carboxylate: ##STR00121## (E)-4-oxo-4-phenylbut-2-en-2-yl 1-methyl-1H-indazole-3-carboxylate: ##STR00122## (E)-4-oxo-4-phenylbut-2-en-2-yl 1 benzo[b]thiophene-2-carboxylate: ##STR00123## (E)-4-oxo-4-phenylbut-2-en-2-yl cinnamate: ##STR00124## (E)-4-oxo-4-phenylbut-2-en-2-yl propiolate: ##STR00125## (E)-4-oxo-4-phenylbut-2-en-2-yl 3-phenylpropiolate: ##STR00126## (E)-4-oxo-4-phenylbut-2-en-2-yl 2-phenyl acetate: ##STR00127## (E)-4-oxo-4-phenylbut-2-en-2-yl 2-phenylacetate: ##STR00128## (E)-4-oxo-4-phenylbut-7-en-7-yl 2-phenylacetate: ##STR00129## (E)-4-oxo-4-phenylbut-2-en-2-yl 2-phenylacetate: ##STR00130## (E)-4-(4-bromophenyl)-4-oxobut-2-en-2-yl 4-methoxybenzoate: ##STR00131## (E)-4-(4-nitrophenyl)-4-oxobut-2-en-2-yl 4-methoxybenzoate: ##STR00132## (E)-4-(2-nitrophenyl)-4-oxobut-2-en-2-yl 4-methoxybenzoate: ##STR00133## (E)-4-(3,5-dimethoxyphenyl)-4-oxobut-2-en-2-yl 4-methoxybenzoate: ##STR00134## (E)-4-oxo-5-phenylpent-2-en-2-yl 4-methoxybenzoate: ##STR00135## (E)-4-(4-methoxyphenyl)-4-oxobut-2-en-2-yl 4-methoxybenzoate: ##STR00136## (E)-4-(furan-2-yl)-4-oxobut-2-en-2-yl 4-methoxybenzoate: ##STR00137## (E)-4-oxo-4-(thiophen-2-yl)but-2-en-2-yl 4-methoxybenzoate: ##STR00138## (E)-3-methyl-4-oxo-4-phenylbut-2-en-2-yl 4-methoxybenzoate: ##STR00139## (E)-4-oxo-4-(perfluorophenyl)but-2-en-2-yl 4-methoxybenzoate: ##STR00140## (E)-1-oxo-1-phenylpent-2-en-3-yl 4-methoxybenzoate: ##STR00141## (E)-4-oxo-4-phenylbut-2-en-2-yl 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanoate: ##STR00142## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alaninate: ##STR00143## (E)-4-oxo-4-phenylbut-2-en-2-yl ((benzyloxy)carbonyl)-L-alaninate: ##STR00144## (E)-4-oxo-4-phenylbut-2-en-2-yl ((benzyloxy)carbonyl)-L-serinate: ##STR00145## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-threoninate: ##STR00146## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-tryptophanate: ##STR00147## (E)-4-oxo-4-phenylbut-2-en-2-yl (tert-butoxycarbonyl)-L-threoninate: ##STR00148## (E)-4-oxo-4-phenylbut-2-en-2-yl (tert-butoxycarbonyl)-L-phenylalaninate: ##STR00149## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-leucinate: ##STR00150## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)glycinate: ##STR00151## (E)-4-(tert-butyl)1-(4-oxo-4-phenylbut-2-en-2-yl) (((9H-fluoren-9-yl)methoxy)carbonyl)-L-aspartate: ##STR00152## (E)-4-oxo-4-phenylbut-2-en-2-yl N-(((9H-fluoren-9-yl)methoxy)carbonyl)-S-trityl-L-cysteinate: ##STR00153## (E)-4-oxo-4-phenylbut-2-en-2-yl(5)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-phenylacetate: ##STR00154## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-methioninate: ##STR00155## (E)-4-oxo-4-phenylbut-2-en-2-yl N.sup.2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N.sup.6-(tert-butoxycarbonyl)-L-lysinate: ##STR00156## (E)-4-oxo-4-phenylbut-2-en-2-yl N.sup.2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N.sup.4-trityl-L-asparaginate: ##STR00157## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-isoleucinate: ##STR00158## tert-butyl (S,E)-4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-oxo-3-((4-oxo-4-phenylbut-2-en-2-yl)oxy)propyl)-1H-imidazole-1-carboxylate: ##STR00159## (E)-4-oxo-4-phenylbut-2-en-2-yl (5)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxy)phenyl)propanoate: ##STR00160## (E)-1-((9H-fluoren-9-yl)methyl) 2-(4-oxo-4-phenylbut-2-en-2-yl) (S)-pyrrolidine-1,2-dicarboxylate: ##STR00161## (E)-4-oxo-4-phenylbut-2-en-2-yl N.sup.2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N.sup.5-trityl-L-glutaminate: ##STR00162## (E)-4-oxo-4-phenylbut-2-en-2-yl (Z)—N.sup.ω,N.sup.ω′-bis((benzyloxy)carbonyl)-N.sup.2-(tert-butoxycarbonyl)-L-argininate: ##STR00163## (E)-4-oxo-4-phenylbut-2-en-2-yl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-phenylalaninate: ##STR00164## (L)-4-oxo-4-phenylbut-2-en-2-yl ((benzyloxy)carbonyl)-L-valyl-L-phenylalaninate: ##STR00165## (E)-4-oxo-4-phenylbut-2-en-2-yl N-(((benzyloxy)carbonyl)-L-leucyl)-O-(tert-butyl)-L-serinate: ##STR00166##

6. A method for preparing an α-carbonyl alkenyl ester having the general formula (I): ##STR00167## comprising the following steps: A) reacting the allenone having the general formula (II) with the carboxylic acid having the general formula (III) in a solvent to obtain the target compound: ##STR00168## wherein R.sup.1 is selected from one of C1-C24 hydrocarbyl, C1-C24 hydrocarbyl substituted by a substituent(s), protected α-amino C1-C12 hydrocarbyl, protected βi-amino C2-C20 hydrocarbyl, protected γ-amino C3-C20 hydrocarbyl, and protected polypeptide chain C1-C20 hydrocarbyl; R.sup.2 is selected from one of aryl, aryl substituted by a substituent(s), heteroaryl and heteroaryl substituted by a substituent(s); and R.sup.3, R.sup.4 and R.sup.5 are the same or different, and are each independently selected from one of H, C1-C18 hydrocarbyl, C1-C18 hydrocarbyl substituted by a substituent(s), C1-C16 acyl, cyano, halogen or C1-C16 hydrocarbyloxy-carbonyl.

7. The method as claimed in claim 6, wherein R.sup.1 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-bund, cyclopentyl, cyclohexyl, butenyl, adamantyl, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, phenylethynyl, phenyl, naphthyl, anthracen I, phenanthryl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, pyrrolyl, indolyl, indolylmethyl, indazolyl, furyl, benzofuranyl, thienyl, benzothienyl, styryl, phenylethynyl, benzyl 11-hydroxyundecyl, pentadecyl, protected α-amino C1-C20 alkyl, protected β-amino C2-C20 alkyl, protected γ-amino C3-C20 alkyl, and protected polypeptide chain C1-C20 alkyl; and/or R.sup.2 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 2-methylphenyl, 2-methoxyphenyl, 3,5-dimethoxyphenyl, 3-nitrophenyl, 4-nitrophenyl, 2,4-dinitrophenyl, 3,5-dinitrophenyl, pentafluorophenyl, 4-trifluoromethylphenyl, 3,5-dichlorophenyl, 1-naphthyl, 2-naphthyl, furanyl and thienyl; and/or R.sup.3, R.sup.4, R.sup.5 are the same or different from each other, and are each independently selected from one of H, methyl, formyl, acetyl, propionyl, butyryl, cyano, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl and benzyloxycarbonyl.

8. (canceled)

9. The method as claimed in claim 6, wherein, in step A), the molar ratio of the allenone having the general formula (II) to the carboxylic acid having the general formula (III) is 1:1-2; and/or step A) comprises: adding the allenone having the general formula (II) and the solvent together into a reactor in proportion, and then adding the carboxylic acid having the general formula (III), reacting at 0-100° C. with stirring for 0.5-320 h, using TLC to monitor the end of the reaction, and obtaining the α-carbonyl alkenyl ester having the general formula (I) by column chromatography from the reaction mixture after the completion of the reaction.

10. (canceled)

11. A method for preparing the amides having the general formula (V) using α-carbonyl alkenyl ester as intermediates, comprising the following steps: B) reacting the α-carbonyl alkenyl ester compound having the general formula (I) and the compound having the general formula (IV) in a solvent, and optionally separating the reaction products, to obtain the target compound (V): ##STR00169## wherein R.sup.1 is selected from one of C1-C24 hydrocarbyl, C1-C24 hydrocarbyl substituted by a substituent(s), protected α-amino C1-C20 hydrocarbyl, protected β-amino C2-C20 hydrocarbyl, protected γ-amino C3-C20 hydrocarbyl, and protected polypeptide chain C1-C20 hydrocarbyl; R.sup.2 is selected from one of aryl, aryl substituted by a substituent(s), heteroaryl and heteroaryl substituted by a substituent(s); R.sup.3, R.sup.4, R.sup.5 are the same or different, and are each independently selected from one of H, C1-C18 hydrocarbyl, C1-C18 hydrocarbyl substituted by a substituent(s), C1-C16 acyl, cyano, halogen or C1-C16 hydrocarbyloxy-carbonyl; and R.sup.6 and R.sup.7 are each independently selected from one of H, amino, C1-C24 hydrocarbyl, C1-C24 hydrocarbyl substituted by a substituent(s), or amino oligomers or amino polymers with primary and/or secondary amine group(s); or, R.sup.6 and R.sup.7 together with the N atom to which they are attached form a C3-C24 cyclic ring.

12. The method as claimed in claim 11, wherein: R.sup.1 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, butenyl, adamantyl, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, phenylethynyl, phenyl, naphthyl, anthracenyl, phenanthryl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, pyrrolyl, indolyl, indolylmethyl, indazolyl, furyl, benzofuranyl, thienyl, benzothienyl, styryl, phenylethynyl, benzyl, 11-hydroxyundecyl, pentadecyl, protected α-amino C1-C20 alkyl, protected β-amino C2-C20 alkyl, protected γ-amino C3-C20 alkyl, and protected polypeptide chain C1-C20 alkyl; and/or R.sup.2 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 2-methylphenyl, 2-methoxyphenyl, 3,5-dimethoxyphenyl, 3-nitrophenyl, 4-nitrophenyl, 2,4-dinitrophenyl, 3,5-dinitrophenyl, pentafluorophenyl, 4-trifluoromethylphenyl, 3,5-dichlorophenyl, 1-naphthyl, 2-naphthyl, furanyl and thienyl; and/or R.sup.3, R.sup.4, R.sup.5 are the same or different from each other, and are each independently selected from one of H, methyl, formyl, acetyl, propionyl, butyryl, cyano, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl and benzyloxycarbonyl; and/or R.sup.6 and R.sup.7 are each independently selected from one of H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, ethanolylphenyl, phenyl, phenethyl naphthyl, 3-indoleethyl, α-acyl C1-C20 alkyl, β-acyl C2-C20 alkyl, γ-acyl C3-C20 alkyl, polypeptide chain C1-C20 alkyl.

13. (canceled)

14. The method as claimed in claim 12, wherein, in step B), the molar ratio of the α-carbonyl alkenyl ester having the general formula (I) to the compound having the general formula (IV) is 1:1-2; and/or step B) comprises: adding the α-carbonyl alkenyl ester having the general formula (I) and the solvent together in a molar proportion to a reactor, and then adding the compound having the general formula (IV), reacting at −40˜100° C. with stirring for 0.05-24 h, using TLC to monitor the completion of the reaction, and obtaining the amide having the general formula (V) by column chromatography after the completion of the reaction.

15. A method for preparing an amide having the general formula (V), comprising: C) reacting the allenone compound having the general formula (II) with the carboxylic acid having the general formula (III) in a first solvent, optionally separating the reaction product, and then obtaining the α-carbonylalkenyl ester; and D) reacting the α-carbonylalkenyl ester prepared in step C) with a compound having general formula (IV) in a second solvent, optionally separating the reaction product, to obtain an amide having the general formula (V): ##STR00170## wherein R.sup.1 is selected from one of C1-C24 hydrocarbyl, C1-C24 hydrocarbyl substituted by a substituent(s), protected α-amino C1-C20 hydrocarbyl, protected β-amino C2-C20 hydrocarbyl, protected γ-amino C3-C20 hydrocarbyl, and protected polypeptide chain C1-C20 hydrocarbyl; R.sup.2 is selected from one of aryl, aryl substituted by a substituent(s), heteroaryl and heteroaryl substituted by a substituent(s); R.sup.3, R.sup.4, R.sup.5 are the same or different, and are each independently selected from one of H, C1-C18 hydrocarbyl, C1-C18 hydrocarbyl substituted by a substituent(s), C1-C16 acyl, cyano, halogen or C1-C16 hydrocarbyloxy-carbonyl; and R.sup.6 and R.sup.7 are each independently selected from one of H, amino, C1-C24 hydrocarbyl, C1-C24 hydrocarbyl substituted by a substituent(s), or amino oligomers or amino polymers with primary and/or secondary amines; or, R.sup.6 and R.sup.7 together with the N atom to which they are attached form a C3-C24 cyclic ring; wherein the first solvent is the same or different from the second solvent.

16. (canceled)

17. The method as claimed in claim 15, wherein, R.sup.1 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, butenyl, adamantyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, phenylethynyl, phenyl, naphthyl, anthracenyl, phenanthryl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, pyrrolyl, indolyl, indolylmethyl, indazolyl, furyl, benzofuranyl, thienyl, benzothienyl, styryl, phenylethynyl, benzyl, 11-hydroxyundecyl, pentadecyl, protected α-amino C1-C20 alkyl, protected β-amino C2-C20 alkyl, protected γ-amino C3-C20 alkyl, and protected polypeptide chain C1-C20 alkyl; and/or R.sup.2 is selected from one of methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 2-methylphenyl, 2-methoxyphenyl, 3,5-dimethoxyphenyl, 3-nitrophenyl, 4-nitrophenyl, 2,4-dinitrophenyl, 3,5-dinitrophenyl, pentafluorophenyl, 4-trifluoromethylphenyl, 3,5-dichlorophenyl, 1-naphthyl, 2-naphthyl, furanyl and thienyl; and/or R.sup.3, R.sup.4, R.sup.5 are the same or different from each other, and are each independently selected from one of H, methyl, formyl, acetyl, propionyl, butyryl, cyano, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl and benzyloxycarbonyl; and/or R.sup.6 and R.sup.7 are each independently selected from one of H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, ethanolylphenyl, phenyl, phenethyl, naphthyl, 3-indoleethyl, α-acyl C1-C20 alkyl, β-acyl C2-C20 alkyl, γ-acyl C3-C20 alkyl, polypeptide chain C1-C20 alkyl.

18. The solid-phase synthesis method of polypeptide chain using the α-carbonyl alkenyl ester having the general formula (I) described in claim 1, comprising: 1) coupling a resin solid phase carrier with the terminal amino acid of the target polypeptide chain sequence or the α-carbonyl alkenyl ester corresponding to the terminal amino acid in a solvent in the presence of a base or a catalyst to obtain PG-AA-resin; 2) according to the sequence of the target polypeptide chain to be synthesized, sequentially adding the α-carbonyl alkenyl ester corresponding to the sequence amino acids and the catalyst or base, and coupling in a solvent to obtain the PG-AA-AA-resin; 3) deprotecting and cleaving the PG-AA-AA-resin, and removing the side chain protecting group and the resin to obtain crude peptide; 4) purifying and freeze-drying the crude peptide to obtain the target polypeptide chain.

19. The method as claimed in claim 18, wherein, the resin solid phase carrier in step 1) is selected from one of Marrifield resin, Wang resin, 2-CTC resin, and MBHA resin; and/or the α-carbonyl alkenyl ester is selected from one or more of those having the general formula (I) and wherein R.sup.1 is a protected α-aminoalkyl, a protected β-aminoalkyl, a protected γ-aminoalkyl, or a protected polypeptide chain alkyl; and/or the catalyst is selected from one of HOAt (1-hydroxy-7-azobenzotriazole), HOBt (1-hydroxybenzotriazole), HOOBt (3-hydroxy-1,2,3-benzotriazin-4(3H)-one), HOSu (N-hydroxysuccinimide), COMU ((1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate) and HOPHT (N-hydroxyphthalimide); and/or the base is selected from one or more of 4-dimethylaminopyridine, pyridine, N-methylimidazole, N,N-diisopropylethylamine (DIEA).

20. (canceled)

21. The method as claimed in claim 18, wherein, in step 1), the amount of the catalyst used is 0.3-10 times the molar amount of the resin used as the solid support; and/or the amount of the allenone used is 1-10 times the molar amount of the PG-AA-resin; and/or in step 201), the amount of the deprotection reagent added is 1-100 times the molar amount of the PG-AA-resin; and/or the deprotection reagent is a solution of piperidine in N,N-dimethylformamide (DMF) or a solution of trifluoroacetic acid in dichloromethane (DCM); and/or in step 202), the amount of the α-carbonyl alkenyl ester is 1-10 times the molar amount of the PG-AA-resin; and/or the amount of the catalyst used is 0.3-10 times the molar amount of the PG-AA-resin; and/or the solvent is an aprotic organic solvent; the aprotic organic solvent is selected from one or two of N,N-dimethylformamide (DMF) and dimethylsulfoxide (DMSO); the mass ratio of the amount of the solvent to the amount of the PG-AA-resin is 10-300:1; and/or in step 4), the purification is carried out by reversed high performance liquid chromatography, the chromatography column is a C18 reversed silica gel column, and the mobile phase is water and acetonitrile.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0175] FIG. 1 is HPLC chart of leucine enkephalin crude product;

[0176] FIG. 2 is LC-MS chart of leucine enkephalin product;

[0177] FIG. 3 is HPLC chart of oxytocin linear backbone crude product;

[0178] FIG. 4 is LC-MS chart of the oxytocin linear backbone product.

DETAILED DESCRIPTION OF THE INVENTION

[0179] The technical solutions of the present invention are illustrated below with examples, and the claimed scope of the present invention includes but is not limited to the following embodiments.

Preparation Example 1

[0180] ##STR00056##

[0181] In a clean 4 mL reaction vial, allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) were added, followed by acetic acid (0.22 mmol); the reaction was then carried out at room temperature for 8 h under stirring, TLC is used to monitor the end of the reaction; after the completion of the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 93%.

[0182] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92 (d, J=7.1 Hz, 2H), 7.55 (t, J=7.4 Hz, 1H), 7.46 (t, J=7.6 Hz, 2H), 6.79 (s, 1H), 2.41 (s, 3H), 2.23 (s, 3H).

[0183] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.4, 168.2, 163.7, 138.7, 132.8, 128.6, 128.1, 113.6, 21.3, 19.0 ppm. HRMS m/z (ESI) calcd for C.sub.12H.sub.12NaO.sub.3 (M+Na).sup.+: 227.0679, found: 227.0685.

Preparation Example 2

[0184] ##STR00057##

[0185] In a clean 4 mL reaction vial, allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) were added, followed by adamantanecarboxylic acid (0.22 mmol); then the reaction was carried out at room temperature for 9 h under stirring, TLC is used to monitor the completion of the reaction; after the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 87%.

[0186] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92 (d, J=7.0 Hz, 2H), 7.54 (t, J=7.3 Hz, 1H), 7.45 (t, J=7.5 Hz, 2H), 6.74 (s, 1H), 2.38 (s, 3H), 2.08 (s, 3H), 1.99 (d, J=2.9 Hz, 6H), 1.86-1.68 (m, 6H).

[0187] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.4, 175.1, 164.5, 138.8, 132.7, 128.5, 128.1, 113.3, 41.2, 38.6, 36.4, 27.8, 19.0 ppm.

[0188] HRMS m/z (ESI) calcd for C.sub.21H.sub.25O.sub.3 (M+H).sup.+: 325.1798, found: 325.1830.

Preparation Example 3

[0189] ##STR00058##

[0190] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by 4-chlorobenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 12 h, using TLC to monitor the end of the reaction; after the end of the reaction, the pure product obtained by column chromatography was a colorless oily liquid with a yield of 93%.

[0191] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.05 (d, J=8.6 Hz, 2H), 7.95 (d, J=7.0 Hz, 2H), 7.55 (t, J=7.3 Hz, 1H), 7.51-7.41 (m, 4H), 6.93 (s, 1H), 2.52 (s, 3H).

[0192] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.2, 163.7, 163.2, 140.5, 138.6, 132.9, 131.5, 129.1, 128.6, 128.2, 127.6, 114.0, 19.0 ppm.

[0193] HRMS m/z (ESI) calcd for C.sub.17H.sub.14ClO.sub.3 (M+H).sup.+: 301.0626, found: 301.0598.

Preparation Example 4

[0194] ##STR00059##

[0195] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by 2-furoic acid (0.22 mmol); then the reaction was stirred at room temperature for 8 h, using TLC to monitor the end of the reaction; after the completion of the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 87%.

[0196] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.94 (d, J=7.1 Hz, 2H), 7.67 (dd, J=1.8, 0.9 Hz, 1H), 7.55 (t, J=7.4 Hz, 1H), 7.46 (t, J=7.5 Hz, 2H), 7.34 (dd, J=3.5, 0.8 Hz, 1H), 6.95 (s, 1H), 6.59 (dd, J=3.5, 1.8 Hz, 1H), 2.51 (s, 3H).

[0197] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.2, 163.1, 155.7, 147.4, 143.6, 138.6, 132.9, 128.6, 128.2, 119.9, 113.9, 112.3, 18.9 ppm.

[0198] HRMS m/z (ESI) calcd for C.sub.15H.sub.12NaO.sub.4 (M+Na).sup.+: 279.0628, found: 279.0637.

Preparation Example 5

[0199] ##STR00060##

[0200] To a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by 1-methylindazole-3-carboxylic acid (0.22 mmol); then the reaction was stirred at room temperature for 28 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 90%.

[0201] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.24 (d, J=8.2 Hz, 1H), 7.97 (d, J=7.2 Hz, 2H), 7.61-7.42 (m, 5H), 7.43-7.33 (m, 1H), 7.03 (s, 1H), 4.23 (s, 3H), 2.62 (s, 3H).

[0202] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.3, 163.9, 159.8, 141.2, 138.7, 133.5, 132.9, 128.6, 128.2, 127.3, 124.0, 123.7, 121.9, 114.3, 109.8, 36.7, 19.3 ppm.

[0203] HRMS m/z (ESI) calcd for C.sub.19H.sub.16N.sub.2NaO.sub.3 (M+Na).sup.+: 343.1053, found: 343.1056.

Preparation Example 6

[0204] ##STR00061##

[0205] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by benzothiophene-2-carboxylic acid (0.22 mmol); then the reaction was stirred at room temperature for 12 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with the yield of 85%.

[0206] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.19 (s, 1H), 8.02-7.86 (m, 4H), 7.65-7.34 (m, 5H), 7.00 (s, 1H), 2.55 (s, 3H).

[0207] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.3, 163.4, 160.1, 142.8, 138.6, 138.6, 132.9, 132.2, 132.2, 128.6, 128.2, 127.6, 125.9, 125.2, 122.9, 113.9, 19.0 ppm.

[0208] HRMS m/z (ESI) calcd for C.sub.19H.sub.14NaO.sub.3S (M+Na).sup.+: 345.0556, found: 345.0563.

Preparation Example 7

[0209] ##STR00062##

[0210] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by cinnamic acid (0.22 mmol); then the reaction was stirred at room temperature for 10 h, using TLC to monitor the completion of the reaction; after the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 96%.

[0211] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.94 (d, J=7.0 Hz, 2H), 7.82 (d, J=16.0 Hz, 1H), 7.62-7.50 (m, 3H), 7.48-7.40 (m, 5H), 6.90 (s, 1H), 6.52 (d, J=16.0 Hz, 1H), 2.49 (s, 3H).

[0212] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.4, 164.2, 163.9, 147.2, 138.8, 134.0, 132.8, 131.0, 129.1, 128.6, 128.4, 128.2, 116.9, 113.5, 19.1 ppm.

[0213] HRMS m/z (ESI) calcd for C.sub.19H.sub.16NaO.sub.3 (M+Na).sup.+: 315.0992, found: 315.0999.

Preparation Example 8

[0214] ##STR00063##

[0215] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by propargylic acid (0.22 mmol); then the reaction was stirred at room temperature for 1.5 h, the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 87%.

[0216] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91 (d, J=7.0 Hz, 2H), 7.56 (t, J=7.4 Hz, 1H), 7.46 (t, J=7.6 Hz, 2H), 6.85 (s, 1H), 3.09 (s, 1H), 2.42 (s, 3H);

[0217] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 189.9, 162.2, 149.5, 138.2, 133.1, 128.6, 128.1, 114.3, 76.9, 73.9, 18.5 ppm.

[0218] HRMS m/z (ESI) calcd for C.sub.13H.sub.10NaO.sub.3 (M+Na).sup.+: 237.0528, found: 237.0635.

Preparation Example 9

[0219] ##STR00064##

[0220] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by phenylpropargylic acid (0.22 mmol); then the reaction was stirred at room temperature for 4 h, using TLC to monitor the end of the reaction; after the end of the reaction, the pure product obtained by column chromatography was a colorless oily liquid with a yield of 93%.

[0221] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.93 (d, J=7.0 Hz, 2H), 7.63 (d, J=7.0 Hz, 2H), 7.56 (t, J=7.3 Hz, 1H), 7.54-7.36 (m, 5H), 6.90 (s, 1H), 2.48 (s, 3H).

[0222] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.1, 162.8, 150.9, 138.5, 133.2, 133.0, 131.2, 128.7, 128.6, 128.2, 119.0, 114.2, 88.8, 80.0, 18.8 ppm.

[0223] HRMS m/z (ESI) calcd for C.sub.19H.sub.14NaO.sub.3 (M+Na).sup.+: 313.0835, found: 313.0841.

Preparation Example 10

[0224] ##STR00065##

[0225] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by phenylacetic acid (0.22 mmol); then the reaction was stirred at room temperature for 5 h, using TLC to monitor the completion of the reaction; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 88%.

[0226] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90 (d, J=7.0 Hz, 2H), 7.54 (t, J=7.4 Hz, 1H), 7.45 (t, J=7.7 Hz, 2H), 7.41-7.28 (m, 5H), 6.78 (s, 1H), 3.79 (s, 2H), 2.37 (s, 3H);

[0227] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.3, 168.9, 163.6, 138.6, 132.9, 132.8, 129.2, 128.8, 128.5, 128.1, 127.5, 113.6, 41.5, 18.8 ppm.

[0228] HRMS m/z (ESI) calcd for C.sub.18H.sub.16NaO.sub.3 (M+Na).sup.+: 303.0997, found: 303.1013.

Preparation Example 11

[0229] ##STR00066##

[0230] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Cbz-L-Ser-OH (0.22 mmol); then the reaction was stirred at room temperature for 12 h, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 87%.

[0231] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.89 (d, J=7.6 Hz, 2H), 7.54 (t, J=7.4 Hz, 1H), 7.44 (t, J=7.7 Hz, 2H), 7.37-7.28 (m, 5H), 6.80 (s, 1H), 5.86 (d, J=8.0 Hz, 1H), 5.14 (s, 2H), 4.58 (s, 1H), 4.13 (d, J=7.4 Hz, 1H), 3.99 (d, J=11.3 Hz, 1H), 2.70 (s, 1H), 2.38 (s, 3H).

[0232] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.3, 168.3, 163.2, 156.2, 138.4, 136.0, 133.0, 128.6, 128.6, 128.3, 128.2, 128.1, 114.1, 67.4, 63.0, 56.3, 18.7 ppm.

[0233] HRMS m/z (ESI) calcd for C.sub.21H.sub.21NNaO.sub.6 (M+Na).sup.+: 406.1261, found: 406.1270.

Preparation Example 12

[0234] ##STR00067##

[0235] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Thr-OH (0.22 mmol); then the reaction was stirred at room temperature for 6 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 94%.

[0236] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.88 (d, J=7.6 Hz, 2H), 7.75 (d, J=7.5 Hz, 2H), 7.60 (d, J=7.5 Hz, 2H), 7.53 (t, J=7.2 Hz, 1H), 7.47-7.33 (m, 4H), 7.29 (t, J=7.5 Hz, 2H), 6.80 (s, 1H), 5.76 (s, 1H), 4.46 (t, 4H), 4.23 (t, J=6.9 Hz, 1H), 2.39 (s, 4H), 1.30 (d, J=6.3 Hz, 3H).

[0237] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.2, 168.8, 163.3, 156.8, 143.8, 143.6, 141.3, 138.4, 133.0, 128.6, 128.2, 127.8, 127.1, 125.0, 120.0, 114.0, 68.0, 67.3, 59.4, 47.2, 20.2, 18.8 ppm.

[0238] HRMS m/z (ESI) calcd for C.sub.29H.sub.27NNaO.sub.6 (M+Na).sup.+: 508.1731, found: 508.1740.

Preparation Example 13

[0239] ##STR00068##

[0240] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Trp-OH (0.22 mmol); then the reaction was stirred at room temperature for 12 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 89%.

[0241] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.28 (s, 1H), 7.81 (d, J=7.8 Hz, 2H), 7.78 (d, J=7.6 Hz, 2H), 7.66 (d, J=7.9 Hz, 1H), 7.60-7.50 (m, 3H), 7.48-7.37 (m, 5H), 7.34-7.28 (m, 2H), 7.23 (d, J=7.8 Hz, 1H), 7.16 (t, J=7.5 Hz, 1H), 7.04 (s, 1H), 6.54 (s, 1H), 5.47 (d, J=8.1 Hz, 1H), 4.93-4.83 (m, 1H), 4.52-4.34 (m, 2H), 4.23 (t, J=7.1 Hz, 1H), 3.50-3.30 (m, 2H), 2.30 (s, 3H);

[0242] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.1, 169.7, 163.2, 155.7, 143.7, 143.6, 141.3, 141.2, 138.3, 136.1, 132.9, 128.5, 128.1, 127.7, 127.4, 127.0, 125.0, 123.0, 122.5, 120.0, 118.6, 113.8, 111.4, 109.4, 67.1, 54.8, 47.1, 27.9, 18.6 ppm.

[0243] HRMS m/z (ESI) calcd for C.sub.41H.sub.38N.sub.2NaO.sub.7 (M+Na).sup.+: 693.2571, found: 693.2583.

Preparation Example 14

[0244] ##STR00069##

[0245] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Boc-L-Thr-OH (0.22 mmol); then the reaction was stirred at room temperature for 8 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 92%.

[0246] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90 (d, J=7.2 Hz, 2H), 7.54 (t, J=7.4 Hz, 1H), 7.45 (t, J=7.6 Hz, 2H), 6.81 (s, 1H), 5.40 (d, J=8.8 Hz, 1H), 4.45 (s, 1H), 4.39 (d, J=8.7 Hz, 1H), 2.40 (s, 3H), 2.29 (s, 1H), 1.47 (s, 9H), 1.32 (d, J=6.4 Hz, 3H);

[0247] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.2, 169.1, 163.4, 156.1, 138.4, 132.9, 128.6, 128.2, 113.9, 80.4, 68.0, 59.0, 28.3, 20.2, 18.7 ppm.

[0248] HRMS m/z (ESI) calcd for C.sub.19H.sub.25NNaO.sub.6 (M+Na).sup.+: 386.1850, found: 386.1837.

Preparation Example 15

[0249] ##STR00070##

[0250] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Boc-L-Phe-OH (0.22 mmol), then the reaction was stirred at room temperature for 8 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 92%.

[0251] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.88 (d, J=7.0 Hz, 2H), 7.55 (t, J=7.3 Hz, 1H), 7.45 (t, J=7.6 Hz, 2H), 7.38-7.28 (m, 3H), 7.23 (d, J=6.6 Hz, 2H), 6.64 (s, 1H), 5.05 (d, J=7.6 Hz, 1H), 4.69 (d, J=7.3 Hz, 1H), 3.17 (d, J=6.5 Hz, 2H), 2.33 (s, 3H), 1.44 (s, 9H).

[0252] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.1, 169.7, 163.3, 155.1, 138.5, 135.6, 132.9, 129.4, 128.8, 128.6, 128.2, 127.4, 113.8, 80.4, 54.8, 38.2, 28.3, 18.6 ppm.

[0253] HRMS m/z (ESI) calcd for C.sub.24H.sub.27NNaO.sub.5 (M+Na).sup.+: 432.1781, found: 432.1787.

Preparation Example 16

[0254] ##STR00071##

[0255] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Leu-OH (0.22 mmol); then the reaction was stirred at room temperature for 3 h, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 99%.

[0256] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90 (d, J=7.7 Hz, 2H), 7.75 (d, J=7.5 Hz, 2H), 7.59 (d, J=7.5 Hz, 2H), 7.53 (t, J=7.4 Hz, 1H), 7.43 (t, J=7.6 Hz, 2H), 7.38 (t, J=7.7 Hz, 2H), 7.29 (t, J=7.4 Hz, 2H), 6.79 (s, 1H), 5.27 (d, J=8.6 Hz, 1H), 4.55-4.48 (m, 1H), 4.44 (d, J=5.0 Hz, 2H), 4.23 (t, J=6.9 Hz, 1H), 2.40 (s, 3H), 1.81-1.71 (m, 2H), 1.69-1.59 (m, 1H), 1.00 (d, J=5.7 Hz, 6H).

[0257] .sup.13C NMR (10 MHz, CDCl.sub.3) δ 190.2, 170.8, 163.3, 156.1, 143.8, 143.7, 141.4, 138.5, 133.0, 128.6, 128.2, 127.8, 127.1, 125.0, 120.1, 113.9, 67.1, 52.8, 47.2, 41.2, 24.9, 23.0, 21.7, 18.8 ppm.

[0258] HRMS m/z (ESI) calcd for C.sub.31H.sub.31NNaO.sub.5 (M+Na).sup.+: 520.2094, found: 520.2105.

Preparation Example 17

[0259] ##STR00072##

[0260] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-Gly-OH (0.22 mmol); then the reaction was stirred at room temperature for 36 h, and TLC was used to monitor the completion of the reaction; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 95%.

[0261] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92 (d, J=7.5 Hz, 2H), 7.77 (d, J=7.5 Hz, 2H), 7.61 (d, J=7.4 Hz, 2H), 7.56 (t, J=7.4 Hz, 1H), 7.46 (t, J=7.6 Hz, 2H), 7.41 (t, J=7.4 Hz, 2H), 7.32 (t, J=7.4 Hz, 2H), 6.84 (s, 1H), 5.45 (t, J=6.0 Hz, 1H), 4.46 (d, J=7.0 Hz, 2H), 4.25 (t, J=7.0 Hz, 1H), 4.16 (d, J=5.9 Hz, 2H), 2.42 (s, 3H).

[0262] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.1, 167.6, 162.9, 156.3, 143.6, 141.3, 138.3, 133.0, 128.6, 128.1, 127.7, 127.1, 125.0, 120.0, 113.9, 67.3, 47.0, 42.9, 18.7 ppm.

[0263] HRMS m/z (ESI) calcd for C.sub.27H.sub.23NNaO.sub.5 (M+Na).sup.+: 464.1468, found: 464.1471.

Preparation Example 18

[0264] ##STR00073##

[0265] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Asp(tBu)-OH (0.22 mmol); then the reaction was stirred at room temperature for 6 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 95%.

[0266] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92 (d, J=7.5 Hz, 2H), 7.77 (d, J=7.5 Hz, 2H), 7.62 (dd, J=7.2, 3.0 Hz, 2H), 7.55 (t, J=7.4 Hz, 1H), 7.45 (t, J=7.6 Hz, 2H), 7.40 (t, J=7.5 Hz, 2H), 7.32 (t, J=7.4 Hz, 2H), 6.82 (s, 1H), 5.93 (d, J=8.7 Hz, 1H), 4.83-4.74 (m, 1H), 4.54-4.44 (m, 1H), 4.44-4.35 (m, 1H), 4.27 (t, J=7.1 Hz, 1H), 3.08 (dd, J=17.3, 4.5 Hz, 1H), 2.87 (dd, J=17.3, 4.2 Hz, 1H), 2.42 (s, 3H), 1.50 (s, 9H.

[0267] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 170.1, 168.7, 163.3, 155.9, 143.6, 141.2, 138.4, 132.9, 128.5, 128.1, 127.7, 127.0, 125.0, 119.9, 114.0, 82.3, 67.3, 50.6, 47.0, 37.8, 28.0, 18.5 ppm.

[0268] HRMS m/z (ESI) calcd for C.sub.33H.sub.35NO.sub.7 (M+H).sup.+: 556.2330, found: 556.2325.

Preparation Example 19

[0269] ##STR00074##

[0270] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Cys(Trt)-OH (0.22 mmol); then the reaction was stirred at room temperature for 6 h, and the end of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 85%.

[0271] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91 (d, J=7.7 Hz, 2H), 7.79 (dd, J=7.7, 3.0 Hz, 2H), 7.63 (d, J=6.7 Hz, 2H), 7.56 (t, J=7.4 Hz, 1H), 7.50-7.39 (m, 11H), 7.37-7.28 (m, 8H), 7.23 (d, J=7.1 Hz, 2H), 6.82 (s, 1H), 5.35 (d, J=8.2 Hz, 1H), 4.54-4.36 (m, 3H), 4.26 (t, J=7.0 Hz, 1H), 2.85 (dd, J=12.6, 6.4 Hz, 1H), 2.71 (dd, J=12.6, 4.6 Hz, 1H), 2.40 (s, 3H);

[0272] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 168.2, 163.1, 155.5, 144.0, 143.7, 143.6, 141.2, 141.2, 138.3, 132.9, 129.4, 128.5, 128.1, 128.1, 127.7, 127.1, 127.0, 125.0, 125.0, 120.0, 113.8, 67.2, 67.2, 53.1, 47.0, 33.7, 18.6 ppm.

[0273] HRMS m/z (ESI) calcd for C.sub.47H.sub.39NO.sub.5S (M+H).sup.+: 752.2447, found: 752.2450.

Preparation Example 20

[0274] ##STR00075##

[0275] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Phg-OH (0.22 mmol); then the reaction was stirred at room temperature for 3 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 100%.

[0276] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.88 (d, J=7.7 Hz, 2H), 7.77 (d, J=7.5 Hz, 2H), 7.61 (d, J=7.5 Hz, 2H), 7.55 (t, J=7.4 Hz, 1H), 7.48-7.38 (m, 9H), 7.31 (t, J=7.6 Hz, 2H), 6.75 (s, 1H), 5.87 (d, J=7.2 Hz, 1H), 5.53 (d, J=7.2 Hz, 1H), 4.62-4.34 (m, 2H), 4.24 (t, J=7.1 Hz, 1H), 2.32 (s, 3H).

[0277] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 168.5, 163.1, 155.4, 143.7, 143.6, 141.2, 138.3, 135.2, 132.9, 129.3, 129.1, 128.5, 128.1, 127.7, 127.3, 127.0, 125.0, 120.0, 113.7, 67.3, 58.3, 47.1, 18.4 ppm.

[0278] HRMS m/z (ESI) calcd for C.sub.33H.sub.27NNaO.sub.5 (M+Na).sup.+: 540.1781, found: 540.1772.

Preparation Example 21

[0279] ##STR00076##

[0280] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Fmoc-L-Met-OH (0.22 mmol), then, the reaction was stirred at room temperature for 3 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 85%.

[0281] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91 (d, J=7.7 Hz, 2H), 7.77 (d, J=7.5 Hz, 2H), 7.61 (d, J=7.5 Hz, 2H), 7.56 (t, J=7.4 Hz, 1H), 7.45 (t, J=7.6 Hz, 2H), 7.40 (t, J=7.5 Hz, 2H), 7.32 (t, J=7.4 Hz, 2H), 6.82 (s, 1H), 5.51 (d, J=8.1 Hz, 1H), 4.76-4.58 (m, 1H), 4.47 (d, J=6.9 Hz, 2H), 4.25 (t, J=6.8 Hz, 1H), 2.61 (t, J=7.3 Hz, 2H), 2.41 (s, 3H), 2.37-2.20 (m, 1H), 2.17-2.01 (m, 4H).

[0282] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 169.7, 163.0, 155.9, 143.7, 143.5, 141.3, 138.3, 133.0, 128.6, 128.1, 127.7, 127.0, 124.9, 120.0, 113.9, 67.1, 53.3, 47.1, 31.3, 29.9, 18.7, 15.5 ppm.

[0283] HRMS m/z (ESI) calcd for C.sub.30H.sub.29NNaO.sub.5S (M+Na).sup.+: 538.1659, found: 538.1654.

Preparation Example 22

[0284] ##STR00077##

[0285] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Lys(Boc)-OH (0.22 mmol); then the reaction was stirred at room temperature for 6 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a pale yellow oily liquid with a yield of 98%.

[0286] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90 (d, J=7.8 Hz, 2H), 7.75 (d, J=7.5 Hz, 2H), 7.61 (d, J=7.5 Hz, 2H), 7.53 (t, J=7.4 Hz, 1H), 7.45 (t, J=7.6 Hz, 2H), 7.39 (t, J=7.7 Hz, 2H), 7.30 (t, J=7.5 Hz, 2H), 6.80 (s, 1H), 5.66 (d, J=7.9 Hz, 1H), 4.69 (s, 1H), 4.51-4.34 (m, 3H), 4.23 (t, J=6.9 Hz, 1H), 3.14 (s, 2H), 2.40 (s, 3H), 2.11-1.72 (m, 3H), 1.60-1.48 (m, 3H), 1.44 (s, 9H).

[0287] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 170.1, 163.1, 156.1, 156.1, 143.7, 143.6, 141.2, 138.3, 132.9, 128.5, 128.1, 127.7, 127.0, 125.0, 119.9, 113.8, 79.2, 67.1, 54.0, 47.1, 39.7, 31.4, 29.6, 28.3, 22.4, 18.7 ppm.

[0288] HRMS m/z (ESI) calcd for C.sub.36H.sub.40N.sub.2NaO.sub.7 (M+Na).sup.+: 635.2728, found: 635.2722.

Preparation Example 23

[0289] ##STR00078##

[0290] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Fmoc-L-Asn(Trt)-OH (0.22 mmol); then the reaction was stirred at room temperature for 8 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a pale yellow oily liquid with a yield of 93%.

[0291] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.82 (d, J=7.7 Hz, 2H), 7.73 (t, J=6.3 Hz, 2H), 7.61-7.56 (m, 2H), 7.51 (t, J=7.4 Hz, 1H), 7.42-7.34 (m, 4H), 7.31-7.07 (m, 17H), 6.81 (s, 1H), 6.62 (s, 1H), 6.15 (d, J=8.9 Hz, 1H), 4.82-4.70 (m, 1H), 4.51-4.39 (m, 1H), 4.39-4.28 (m, 1H), 4.22 (t, J=7.0 Hz, 1H), 3.19 (dd, J=16.3, 3.6 Hz, 1H), 2.91 (dd, J=16.3, 3.2 Hz, 1H), 2.24 (s, 3H).

[0292] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.2, 169.1, 168.8, 163.6, 156.1, 144.1, 143.7, 143.5, 141.2, 138.3, 132.8, 128.5, 128.4, 128.2, 128.0, 127.7, 127.2, 127.0, 125.0, 119.9, 113.9, 71.0, 67.2, 50.8, 47.0, 38.7, 18.5 ppm.

[0293] HRMS m/z (ESI) calcd for C.sub.48H.sub.40N.sub.2NaO.sub.6 (M+Na).sup.+: 763.2779, found: 763.2763.

Preparation Example 24

[0294] ##STR00079##

[0295] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Fmoc-L-Ile-OH (0.22 mmol), then, the reaction was stirred at room temperature for 7 h, and the end of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid, and the yield was 99%.

[0296] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92 (d, J=7.0 Hz, 2H), 7.77 (d, J=7.5 Hz, 2H), 7.62 (d, J=7.4 Hz, 2H), 7.55 (t, J=7.4 Hz, 1H), 7.46 (t, J=7.6 Hz, 2H), 7.40 (t, J=7.6 Hz, 2H), 7.32 (t, J=7.4 Hz, 2H), 6.81 (s, 1H), 5.43 (d, J=9.0 Hz, 1H), 4.51 (dd, J=8.9, 5.1 Hz, 1H), 4.46 (d, J=7.0 Hz, 2H), 4.25 (t, J=6.9 Hz, 1H), 2.43 (s, 3H), 2.13-1.96 (m, 1H), 1.63-1.45 (m, 1H), 1.39-1.20 (m, 1H), 1.06 (d, J=6.8 Hz, 2H), 1.01 (t, J=7.4 Hz, 3H), 0.96-0.87 (m, 1H);

[0297] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 169.7, 163.0, 156.1, 143.7, 143.6, 141.2, 138.4, 132.9, 128.5, 128.1, 127.7, 127.0, 124.9, 119.9, 113.9, 67.0, 58.5, 47.1, 37.8, 25.1, 18.7, 15.6, 11.5 ppm.

[0298] HRMS m/z (ESI) calcd for C.sub.31H.sub.31NNaO.sub.5 (M+Na).sup.+: 520.2100, found: 520.2128.

Preparation Example 25

[0299] ##STR00080##

[0300] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-His(Boc)-OH (0.22 mmol); then the reaction was stirred at room temperature for 12 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 24%.

[0301] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.06 (s, 1H), 7.91 (d, J=7.5 Hz, 1H), 7.76 (d, J=7.5 Hz, 2H), 7.62 (t, J=7.4 Hz, 2H), 7.55 (t, J=7.3 Hz, 1H), 7.45 (t, J=7.6 Hz, 2H), 7.40 (t, J=7.5 Hz, 2H), 7.35-7.27 (m, 2H), 7.25 (s, 1H), 6.85 (s, 1H), 6.33 (d, J=8.1 Hz, 1H), 4.90-4.72 (m, 1H), 4.49-4.33 (m, 2H), 4.27 (t, J=7.3 Hz, 1H), 3.28 (dd, J=15.0, 5.3 Hz, 1H), 3.17 (dd, J=15.0, 4.8 Hz, 1H), 2.39 (s, 3H), 1.60 (s, 9H);

[0302] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.1, 169.1, 163.4, 156.0, 146.7, 143.8, 143.7, 141.2, 138.5, 138.0, 137.1, 132.8, 128.5, 128.1, 127.7, 127.0, 125.1, 119.9, 114.9, 114.0, 85.9, 67.3, 53.7, 47.1, 29.7, 27.8, 18.7 ppm.

[0303] HRMS m/z (ESI) calcd for C.sub.36H.sub.35N.sub.3NaO.sub.7 (M+Na).sup.+: 644.2373, found: 644.2402.

Preparation Example 26

[0304] ##STR00081##

[0305] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Fmoc-L-Tyr(tBu)-OH (0.22 mmol); then the reaction was stirred at room temperature for 12 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a pale yellow oily liquid with a yield of 85%.

[0306] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.89 (d, J=7.7 Hz, 2H), 7.78 (d, J=7.5 Hz, 2H), 7.60 (d, J=7.5 Hz, 2H), 7.55 (d, J=7.4 Hz, 1H), 7.46 (t, J=7.6 Hz, 2H), 7.41 (t, J=7.6 Hz, 2H), 7.32 (t, J=7.4 Hz, 2H), 7.11 (d, J=8.0 Hz, 2H), 6.97 (d, J=7.9 Hz, 2H), 6.66 (s, 1H), 5.38 (s, 1H), 4.77 (dd, J=6.9 Hz, 2H), 4.57-4.36 (m, 2H), 4.24 (t, J=6.8 Hz, 1H), 3.17 (dd, J=6.4, 3.0 Hz, 2H), 2.34 (s, 3H), 1.34 (s, 9H).

[0307] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 189.9, 169.3, 163.0, 155.5, 154.8, 143.7, 143.6, 141.3, 138.3, 132.9, 129.8, 128.6, 128.1, 127.7, 127.0, 125.0, 124.9, 124.2, 120.0, 113.8, 78.5, 67.0, 55.1, 47.1, 37.5, 28.8, 18.6 ppm.

[0308] HRMS m/z (ESI) calcd for C.sub.38H.sub.37NNaO.sub.6 (M+Na).sup.+: 626.2513, found: 626.2505.

Preparation Example 27

[0309] ##STR00082##

[0310] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Pro-OH (0.22 mmol); then the reaction was stirred at room temperature for 8 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 98%.

[0311] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92 (d, J=7.7 Hz, 1H), 7.76 (t, J=7.0 Hz, 3H), 7.66-7.58 (m, 2H), 7.56-7.48 (m, 1H), 7.45-7.29 (m, 6H), 6.89-6.63 (m, 1H), 4.54-4.37 (m, 3H), 4.31-4.21 (m, 1H), 3.74-3.66 (m, 2H), 3.58 (dd, J=10.3, 7.3 Hz, 2H), 2.47-2.31 (m, 4H), 2.19-2.10 (m, 1H), 2.08-1.93 (m, 2H).

[0312] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.2, 189.9, 170.1, 170.0, 163.4, 163.1, 154.8, 154.2, 143.9, 143.9, 143.7, 143.5, 141.2, 141.2, 138.5, 138.3, 132.8, 132.8, 128.5, 128.1, 128.0, 127.7, 127.6, 127.1, 127.0, 127.0, 125.0, 125.0, 124.9, 119.9, 119.9, 113.6, 113.4, 67.6, 67.5, 59.4, 58.8, 47.3, 47.1, 47.0, 46.5, 31.0, 29.8, 24.5, 23.4, 18.7, 18.6 ppm.

[0313] HRMS m/z (ESI) calcd for C.sub.30H.sub.27NNaO.sub.5 (M+Na).sup.+: 504.1781, found: 504.1771.

Preparation Example 28

[0314] ##STR00083##

[0315] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Fmoc-L-Gln(Trt)-OH (0.22 mmol); then the reaction was stirred at room temperature for 8 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a pale yellow oily liquid with a yield of 93%.

[0316] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.84 (d, J=7.7 Hz, 2H), 7.70 (d, J=7.5 Hz, 2H), 7.57 (d, J=7.5 Hz, 2H), 7.50 (t, J=7.4 Hz, 1H), 7.45-7.31 (m, 4H), 7.30-7.14 (m, 17H), 6.85 (s, 1H), 6.75 (s, 1H), 5.70 (s, 1H), 4.52-4.45 (m, 1H), 4.44-4.36 (m, 2H), 4.21 (t, J=6.9 Hz, 1H), 2.46-2.19 (m, 6H), 2.14-1.92 (m, 1H);

[0317] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 170.6, 169.6, 163.1, 156.2, 144.4, 143.7, 143.5, 141.2, 141.2, 138.3, 132.8, 128.6, 128.5, 128.1, 127.9, 127.7, 127.0, 125.0, 119.9, 113.9, 70.7, 66.9, 53.7, 47.1, 32.8, 26.9, 18.6 ppm.

[0318] HRMS m/z (ESI) calcd for C.sub.49H.sub.42N.sub.2NaO.sub.6 (M+Na).sup.+: 777.2941, found: 777.2905.

Preparation Example 29

[0319] ##STR00084##

[0320] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE), followed by Boc-L-Arg(Cbz).sub.2-OH (0.22 mmol); then the reaction was stirred at room temperature for 10 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a light yellow oily liquid with a yield of 95%.

[0321] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.45 (s, 1H), 9.26 (s, 1H), 7.89 (d, J=8.4 Hz, 2H), 7.58-7.50 (m, 1H), 7.44 (t, J=7.8 Hz, 2H), 7.42-7.34 (m, 6H), 7.34-7.29 (m, 2H), 7.28-7.22 (m, 2H), 6.74 (s, 1H), 5.37 (d, J=8.3 Hz, 1H), 5.24 (s, 2H), 5.12 (s, 2H), 4.39 (s, 2H), 4.02 (s, 2H), 2.33 (s, 3H), 1.95-1.82 (m, 2H), 1.80-1.70 (m, 3H), 1.44 (s, 9H).

[0322] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 170.2, 163.7, 163.4, 160.4, 155.6, 155.4, 138.4, 136.7, 134.5, 132.8, 128.8, 128.8, 128.5, 128.3, 128.2, 128.1, 127.8, 127.7, 113.7, 80.0, 68.9, 66.9, 53.5, 44.0, 28.6, 28.2, 24.9, 18.6 ppm.

[0323] HRMS m/z (ESI) calcd for C.sub.37H.sub.43N.sub.4O.sub.9 (M+H).sup.+: 687.3025, found: 687.3030.

Preparation Example 30

[0324] ##STR00085##

[0325] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by Cbz-L-Leu-L-Ser(tBu)-OH (0.22 mmol); then the reaction was stirred at room temperature for 10 h, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a pale yellow oily liquid with a yield of 99%.

[0326] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.93-7.88 (m, 2H), 7.55 (t, J=7.4 Hz, 1H), 7.45 (t, J=7.7 Hz, 2H), 7.37-7.27 (m, 5H), 6.79-6.72 (m, 2H), 5.28 (d, J=9.2 Hz, 1H), 5.11 (s, 2H), 4.85 (dt, J=8.5, 3.0 Hz, 1H), 4.34-4.22 (m, 1H), 3.93 (dd, J=9.1, 3.0 Hz, 1H), 3.61 (dd, J=9.0, 3.0 Hz, 1H), 2.40 (s, 3H), 1.78-1.65 (m, 2H), 1.61-1.50 (m, 1H), 1.18 (s, 9H), 0.95 (d, J=6.1 Hz, 6H).

[0327] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.1, 172.1, 167.9, 163.4, 156.0, 138.5, 136.2, 132.9, 128.6, 128.5, 128.1, 128.1, 128.0, 113.8, 73.7, 67.0, 61.9, 53.4, 52.9, 41.6, 27.3, 24.6, 22.8, 22.0, 18.7 ppm.

[0328] HRMS m/z (ESI) calcd for C.sub.31H.sub.41N.sub.2O.sub.7 (M+H).sup.+: 553.2908, found: 553.2902.

Preparation Example 31

[0329] ##STR00086##

[0330] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by 4-methoxybenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 15 h, TLC was used to monitor the end of the reaction; after the end of the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 94%.

[0331] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.07 (d, J=8.9 Hz, 2H), 7.98 (dd, J=8.7, 5.6 Hz, 2H), 7.13 (t, J=8.6 Hz, 2H), 6.98 (d, J=8.9 Hz, 2H), 6.90 (s, 1H), 3.89 (s, 3H), 2.51 (s, 3H).

[0332] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 188.8, 166.9, 164.5, 164.3, 164.2, 163.7, 135.2, 135.1, 132.3, 130.8, 130.7, 121.4, 115.8, 115.5, 114.0, 113.3, 55.5, 19.2 ppm.

[0333] HRMS m/z (ESI) calcd for C.sub.18H.sub.16FO.sub.4 (M+H).sup.+: 315.1027, found: 315.1021.

Preparation Example 32

[0334] ##STR00087##

[0335] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by 4-methoxybenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 11 h, TLC was used to monitor the end of the reaction; after the end of the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 78%.

[0336] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.06 (d, J=8.9 Hz, 2H), 7.89 (d, J=8.6 Hz, 2H), 7.43 (d, J=8.6 Hz, 2H), 6.97 (d, J=9.0 Hz, 2H), 6.89 (s, 1H), 3.89 (s, 3H), 2.52 (s, 3H).

[0337] .sup.13C NMR (10 MHz, CDCl.sub.3) δ 189.1, 164.9, 164.2, 163.7, 139.2, 137.1, 132.3, 129.6, 128.9, 121.3, 114.0, 113.1, 55.5, 19.3 ppm.

[0338] HRMS m/z (ESI) calcd for C.sub.18H.sub.16ClO.sub.4 (M+H).sup.+: 331.0732.

Preparation Example 33

[0339] ##STR00088##

[0340] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by 4-methoxybenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 12 h, TLC was used to monitor the end of the reaction; after the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 78%.

[0341] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.06 (d, J=8.9 Hz, 2H), 7.81 (d, J=8.5 Hz, 2H), 7.60 (d, J=8.5 Hz, 2H), 6.97 (d, J=8.9 Hz, 2H), 6.88 (s, 1H), 3.89 (s, 3H), 2.51 (s, 3H).

[0342] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 189.2, 165.0, 164.2, 163.7, 137.5, 132.3, 131.9, 129.7, 127.9, 121.3, 114.0, 113.1, 55.6, 19.3 ppm.

[0343] HRMS m/z (ESI) calcd for C.sub.18H.sub.16BrO.sub.4 (M+H).sup.+: 375.0226, found: 375.0221.

Preparation Example 34

[0344] ##STR00089##

[0345] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by 4-methoxybenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 96 h, the TLC was used to monitor the end of the reaction; after the reaction, the pure product was obtained by column chromatography as an orange oily liquid with a yield of 59%.

[0346] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.31 (d, J=8.7 Hz, 2H), 8.08 (t, J=8.5 Hz, 2H), 7.13-6.81 (m, 3H), 3.90 (s, 3H), 2.56 (s, 3H).

[0347] .sup.13C NMR (101 MHz, CDCl.sub.3) δ 188.6, 166.6, 164.4, 163.5, 150.1, 143.5, 132.4, 129.1, 123.8, 121.1, 114.1, 112.7, 55.6, 19.6 ppm.

[0348] HRMS m/z (ESI) calcd for C.sub.18H.sub.15NNaO.sub.6 (M+Na).sup.+: 364.0792, found: 361.0789.

Preparation Example 35

[0349] ##STR00090##

[0350] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by 4-methoxybenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 36 h, TLC was used to monitor the end of the reaction; after the end of the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 79%.

[0351] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.06 (d, J=8.9 Hz, 2H), 7.09 (d, J=2.3 Hz, 2H), 6.97 (d, J=8.9 Hz, 2H), 6.85 (s, OH), 6.64 (t, J=2.3 Hz, OH), 3.89 (s, 3H), 3.84 (s, 6H), 2.51 (s, 3H).

[0352] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 190.0, 164.5, 164.1, 163.7, 160.9, 140.8, 132.3, 121.4, 113.9, 113.7, 106.0, 105.3, 55.6, 55.5, 19.2 ppm.

[0353] HRMS m/z (ESI) calcd for C.sub.20H.sub.20NaO.sub.6 (M+Na).sup.+: 379.1152, found: 379.1149.

Preparation Example 36

[0354] ##STR00091##

[0355] In a clean 4 mL reaction vial were added allenone (0.20 mmol) and 2 mL of 1,2-dichloroethane (DCE) followed by 4-methoxybenzoic acid (0.22 mmol); then the reaction was stirred at room temperature for 168 h, and the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a colorless oily liquid with a yield of 88%.

[0356] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.99 (d, J=8.9 Hz, 2H), 7.33 (t, J=7.3 Hz, 2H), 7.29-7.25 (m, 1H), 7.24-7.19 (m, 2H), 6.94 (d, J=8.9 Hz, 2H), 6.24 (s, 1H), 3.87 (s, 3H), 3.76 (s, 2H), 2.43 (s, 3H).

[0357] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 197.0, 164.2, 164.1, 163.6, 134.3, 132.3, 129.6, 128.7, 127.0, 121.3, 115.3, 113.9, 55.5, 51.7, 19.0 ppm.

[0358] HRMS m/z (ESI) calcd for C.sub.19H.sub.18NaO.sub.4 (M+Na).sup.+: 333.1097, found: 333.1095.

Application Example A

[0359] The amide having the general formula (IV) was prepared by using the α-carbonyl alkenyl ester in the general formula (I) as an intermediate.

Example A1

[0360] ##STR00092##

[0361] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 1) (0.20 mmol) and 1.5 mL N,N-dimethylformamide, followed by phenethylamine (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained as a white solid by column chromatography, and the yield was 93%.

[0362] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.27 (t, J=7.3 Hz, 2H), 7.21-7.16 (m, 3H), 6.50 (s, 1H), 3.45 (dd, J=13.3, 7.0 Hz, 2H), 2.79 (t, J=7.2 Hz, 2H), 1.90 (s, 3H).

[0363] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 170.2, 138.7, 128.4, 128.3, 126.1, 40.5, 35.3, 22.8 ppm.

[0364] HRMS m/z (ESI) calcd for C.sub.10H.sub.14NO (M+H).sup.+: 164.1075, found 164.1070.

Example A2

[0365] ##STR00093##

[0366] To a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 7) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by phenethylamine (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the completion of the reaction, the pure product was obtained as a white solid by column chromatography, and the yield was 94%.

[0367] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.62 (d, J=15.6 Hz, 1H), 7.47 (dd, J=6.5, 2.9 Hz, 2H), 7.37-7.29 (m, 5H), 7.28-7.20 (m, 3H), 6.35 (d, J=15.6 Hz, 1H), 5.85 (s, 1H), 3.66 (dd, J=13.0, 6.8 Hz, 2H), 2.89 (t, J=6.9 Hz, 2H).

[0368] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 165.9, 141.0, 138.9, 134.8, 129.6, 128.8, 128.7, 128.6, 127.7, 126.5, 120.7, 40.8, 35.6 ppm.

[0369] HRMS m/z (ESI) calcd for C.sub.17H.sub.18NO (M+H).sup.+: 252.1388, found: 252.1385.

Example A3

[0370] ##STR00094##

[0371] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 8) (0.20 mmol) and N,N-dimethylformamide 1.5 mL, followed by phenethylamine (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained as a white solid by column chromatography, and the yield was 99%.

[0372] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.33 (t, J=7.3 Hz, 2H), 7.27-7.18 (m, 3H), 5.94 (s, 1H), 3.57 (q, J=6.9 Hz, 2H), 2.85 (t, J=7.0 Hz, 2H), 2.75 (s, 1H).

[0373] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 152.1, 138.2, 128.7, 128.7, 126.7, 79.2, 73.1, 40.9, 35.2 ppm.

[0374] HRMS m/z (ESI) calcd for CH.sub.11H.sub.12NO (M+H).sup.+: 174.0919, found: 174.0916.

Example A4

[0375] ##STR00095##

[0376] To a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 6) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by phenethylamine (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the completion of the reaction, the pure product was obtained as a white solid by column chromatography, and the yield was 84%.

[0377] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90-7.75 (m, 2H), 7.68 (s, 1H), 7.42-7.36 (m, 2H), 7.34-7.30 (m, 2H), 7.26-7.24 (m, 3H), 6.26 (s, 1H), 3.71 (dd, J=13.0, 6.8 Hz, 2H), 2.94 (t, J=6.9 Hz, 2H).

[0378] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 162.2, 140.7, 139.0, 138.7, 138.4, 128.8, 128.7, 126.6, 126.3, 125.1, 125.0, 124.9, 122.7, 41.3, 35.7 ppm.

[0379] HRMS m/z (ESI) calcd for C.sub.17H.sub.16NOS (M+H).sup.+: 282.0953, found: 282.0950.

Example A5

[0380] ##STR00096##

[0381] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 9) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by piperidine (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 98%.

[0382] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.59-7.45 (m, 2H), 7.43-7.28 (m, 3H), 3.79-3.72 (m, 2H), 3.64-3.56 (m, 2H), 1.65 (dt, J=9.9, 6.3 Hz, 4H), 1.56 (dt, J=11.0, 5.6 Hz, 2H).

[0383] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 152.8, 132.2, 129.7, 128.4, 120.7, 90.1, 81.4, 48.1, 42.3, 26.4, 25.3, 24.4 ppm.

[0384] HRMS m/z (ESI) calcd for C.sub.14H.sub.16NO (M+H).sup.+: 214.1226, found: 214.1217.

Example A6

[0385] ##STR00097##

[0386] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparation Example 9) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by 2-naphthylamine (0.22 mmol); then the reaction was stirred at room temperature for 12 h, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained as a white solid by column chromatography, and the yield was 99%.

[0387] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.26 (s, 1H), 7.92 (s, 1H), 7.79 (t, J=7.8 Hz, 3H), 7.57 (d, J=6.9 Hz, 2H), 7.55-7.38 (m, 4H), 7.36 (t, J=7.4 Hz, 2H).

[0388] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 151.1, 134.8, 133.8, 132.7, 130.9, 130.4, 129.0, 128.6, 127.8, 127.6, 126.7, 125.4, 119.9, 119.5, 117.1, 85.9, 83.5 ppm.

[0389] HRMS m/z (ESI) calcd for C.sub.19H.sub.14NO (M+H).sup.+: 272.1070, found: 272.1110.

Example A7

[0390] ##STR00098##

[0391] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 9) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by N-methylaniline (0.22 mmol); then, the reaction was stirred at room temperature for 24 h, and TLC was used to monitor the completion of the reaction; after the reaction was completed, the pure product was obtained as a white solid by column chromatography, and the yield was 99%.

[0392] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.50-7.29 (m, 6H), 7.29-7.19 (m, 2H), 7.14 (d, J=6.8 Hz, 2H), 3.40 (s, 3H).

[0393] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 154.3, 143.3, 132.4, 129.9, 129.2, 128.3, 127.9, 127.4, 120.5, 90.9, 82.6, 36.4 ppm.

[0394] HRMS m/z (ESI) calcd for C.sub.16H.sub.14NO (M+H).sup.+: 236.1070, found 236.1085.

Example A8

[0395] ##STR00099##

[0396] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 10) (0.20 mmol) and N,N-dimethylformamide 1.5 mL, followed by aniline (0.22 mmol); then the reaction was stirred at room temperature for 24 h, and the completion of the reaction was monitored by TLC; after the completion of the reaction, the pure product was obtained as a white solid by column chromatography, and the yield was 76%.

[0397] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.40 (t, J=8.5 Hz, 3H), 7.36-7.31 (m, 3H), 7.32-7.23 (m, 2H), 7.17 (s, 1H), 7.08 (t, J=7.4 Hz, 1H), 3.73 (s, 2H).

[0398] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 169.1, 137.6, 134.5, 129.5, 129.2, 128.9, 127.7, 124.5, 119.8, 44.9 ppm.

[0399] HRMS m/z (ESI) calcd for C.sub.14H.sub.14NO (M+H).sup.+: 212.1070, found 212.1058.

Example A9

[0400] ##STR00100##

[0401] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 13) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Thr-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained as a white solid by column chromatography, and the yield was 95%.

[0402] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.20 (s, 1H), 7.74 (d, J=7.5 Hz, 2H), 7.65 (d, J=7.7 Hz, 1H), 7.52-7.47 (m, 2H), 7.38 (t, J=7.4 Hz, 2H), 7.31-7.24 (m, 3H), 7.16 (t, J=7.4 Hz, 1H), 7.10 (t, J=7.4 Hz, 1H), 7.03 (s, 1H), 6.84 (d, J=8.5 Hz, 1H), 5.69 (d, J=7.8 Hz, 1H), 4.61 (d, J=5.8 Hz, 1H), 4.42 (dd, J=8.6, 3.1 Hz, 1H), 4.39-4.25 (m, 2H), 4.21-4.09 (m, 2H), 3.27 (d, J=5.5 Hz, 2H), 2.69 (s, 1H), 1.43 (s, 9H), 1.08 (d, J=5.4 Hz, 3H).

[0403] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 172.0, 169.5, 156.2, 143.8, 143.7, 141.2, 136.2, 127.7, 127.1, 125.1, 123.4, 122.2, 119.9, 119.7, 118.6, 111.3, 110.2, 82.5, 68.4, 67.2, 58.2, 55.7, 47.0, 28.3, 27.9, 19.9 ppm.

[0404] HRMS m/z (ESI) calcd for C.sub.34H.sub.37N.sub.3NaO.sub.6 (M+Na).sup.+: 606.2575, found: 606.2576.

Example A10

[0405] ##STR00101##

[0406] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 14) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Thr-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 15 min, and the reaction was monitored by TLC; after the reaction, the pure product was obtained as a white solid by column chromatography with a yield of 93%.

[0407] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35 (d, J=7.7 Hz, 1H), 5.73 (s, 1H), 4.42 (dd, J=8.7, 2.4 Hz, 1H), 4.35-4.09 (m, 3H), 1.43 (d, J=10.9 Hz, 18H), 1.18 (t, J=6.2 Hz, 6H).

[0408] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.7, 169.8, 156.2, 82.6, 80.3, 68.2, 67.3, 58.7, 58.4, 28.2, 27.9, 20.0, 18.2 ppm.

[0409] HRMS m/z (ESI) calcd for C.sub.17H.sub.32N.sub.2NaO.sub.7 (M+Na).sup.+: 399.2102, found: 399.2094.

Example A11

[0410] ##STR00102##

[0411] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 18) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Tyr(tBu)-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 97%.

[0412] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.75 (d, J=7.5 Hz, 2H), 7.60 (d, J=7.5 Hz, 2H), 7.39 (t, J=7.5 Hz, 2H), 7.30 (t, J=7.3 Hz, 2H), 7.06 (d, J=8.1 Hz, 2H), 6.94-6.77 (m, 3H), 5.79 (d, J=7.9 Hz, 1H), 4.78-4.61 (m, 1H), 4.43-4.32 (m, 2H), 4.28-4.17 (m, 2H), 3.04 (d, J=6.3 Hz, 2H), 2.48-2.28 (m, 2H), 2.14-2.00 (m, 2H), 1.98-1.80 (m, 1H), 1.45 (s, 9H), 1.37 (s, 9H), 1.29 (s, 9H).

[0413] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 172.7, 170.7, 170.1, 156.1, 154.3, 143.8, 143.7, 141.2, 130.8, 129.8, 127.6, 127.0, 125.1, 124.0, 119.9, 82.2, 80.9, 78.2, 67.1, 54.1, 53.9, 47.1, 37.4, 31.6, 28.7, 28.3, 28.0, 27.8 ppm.

[0414] HRMS m/z (ESI) calcd for C.sub.41H.sub.53N.sub.2O.sub.8 (M+H).sup.+: 701.3796, found: 701.3801.

Example A12

[0415] ##STR00103##

[0416] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 19) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Thr(tBu)-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 95%.

[0417] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.76 (t, J=7.0 Hz, 2H), 7.59 (d, J=7.5 Hz, 2H), 7.47 (d, J=7.4 Hz, 6H), 7.45-7.36 (m, 2H), 7.34-7.27 (m, 8H), 7.22 (t, J=7.2 Hz, 3H), 6.56 (d, J=8.8 Hz, 1H), 5.20 (d, J=8.1 Hz, 1H), 4.41-4.33 (m, 2H), 4.29 (d, J=8.6 Hz, 1H), 4.25-4.14 (m, 2H), 3.82-3.71 (m, 1H), 2.79 (dd, J=13.3, 7.9 Hz, 1H), 2.67 (dd, J=13.3, 5.4 Hz, 1H), 1.89 (s, 1H), 1.43 (s, 9H), 1.15 (s, 9H), 1.11 (d, J=6.3 Hz, 3H).

[0418] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 170.2, 169.2, 155.6, 144.3, 143.8, 143.7, 141.2, 129.6, 128.0, 127.6, 127.0, 126.8, 125.1, 119.9, 81.8, 73.8, 67.2, 67.1, 67.0, 58.5, 53.8, 47.0, 34.2, 28.6, 28.0, 20.6 ppm.

[0419] HRMS m/z (ESI) calcd for C.sub.49H.sub.54N.sub.2NaO.sub.6S (M+Na).sup.+: 821.3595, found: 821.3618.

Example A13

[0420] ##STR00104##

[0421] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 23) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Thr-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 90%.

[0422] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.85-7.74 (m, 2H), 7.60 (d, J=7.5 Hz, 2H), 7.43 (t, J=7.5 Hz, 2H), 7.38-7.27 (m, 10H), 7.25-7.17 (m, 6H), 7.10 (s, 1H), 6.54 (d, J=8.3 Hz, 1H), 4.65 (s, 1H), 4.47-4.32 (m, 3H), 4.23 (t, J=7.1 Hz, 2H), 3.14 (dd, J=15.8, 4.5 Hz, 1H), 2.73 (dd, J=15.8, 5.2 Hz, 1H), 2.65 (s, 1H), 2.12-1.97 (m, 1H), 1.49 (s, 9H), 1.14 (d, J=6.4 Hz, 3H).

[0423] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.3, 170.2, 169.3, 156.3, 144.2, 143.7, 143.6, 141.2, 128.6, 127.9, 127.6, 127.0, 125.1, 125.1, 119.9, 82.4, 70.8, 68.3, 67.3, 58.4, 51.7, 47.0, 38.2, 27.9, 20.0 ppm.

[0424] HRMS m/z (ESI) calcd for C.sub.46H.sub.48N.sub.3O.sub.7 (M+H).sup.+: 754.3487, found: 754.3484.

Example A14

[0425] ##STR00105##

[0426] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 24) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Leu-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 90%.

[0427] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.75 (d, J=7.6 Hz, 2H), 7.59 (d, J=7.6 Hz, 2H), 7.39 (t, J=7.5 Hz, 2H), 7.30 (t, J=7.5 Hz, 2H), 6.31 (s, 1H), 5.53 (s, 1H), 4.55-4.30 (m, 3H), 4.21 (t, J=7.1 Hz, 1H), 4.07 (t, J=8.2 Hz, 1H), 1.69-1.54 (m, 3H), 1.53-1.34 (m, 11H), 1.02-0.82 (m, 13H).

[0428] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.6, 170.8, 156.2, 143.8, 143.7, 141.2, 127.6, 127.0, 125.0, 119.9, 119.9, 81.9, 67.0, 59.5, 51.5, 47.1, 41.7, 37.7, 27.9, 24.8, 22.6, 22.1, 15.3, 11.3 ppm.

[0429] HRMS m/z (ESI) calcd for C.sub.31H.sub.43N.sub.2O.sub.5 (M+H).sup.+: 523.3166, found: 523.3160.

Example A15

[0430] ##STR00106##

[0431] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 25) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Leu-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and TLC was used to monitor the end of the reaction; after the reaction, the pure product was obtained as a white solid by column chromatography, and the yield was 85%.

[0432] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.03 (s, 1H), 7.75 (d, J=7.6 Hz, 2H), 7.61 (s, 2H), 7.38 (t, J=7.5 Hz, 2H), 7.30 (t, J=7.4 Hz, 2H), 7.20 (s, 2H), 6.71 (d, J=7.5 Hz, 1H), 4.58 (q, J=6.0 Hz, 1H), 4.46-4.32 (m, 3H), 4.24 (t, J=7.4 Hz, 1H), 3.17 (dd, J=14.8, 4.9 Hz, 1H), 2.97 (dd, J=14.9, 5.8 Hz, 1H), 1.58 (s, 9H), 1.54-1.49 (m, 1H), 1.41 (s, 11H), 0.91-0.74 (m, 6H).

[0433] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.4, 170.4, 156.1, 146.7, 143.8, 141.1, 139.1, 136.6, 127.6, 127.0, 125.1, 119.8, 114.8, 85.6, 81.6, 67.2, 54.6, 51.3, 47.0, 41.4, 30.2, 27.8, 27.7, 24.6, 22.7, 21.7 ppm.

[0434] HRMS m/z (ESI) calcd for C.sub.36H.sub.47N.sub.4O.sub.7 (M+H).sup.+: 647.3439, found: 647.3444.

Example A16

[0435] ##STR00107##

[0436] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 26) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide, followed by H-L-Tyr(tBu)-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 5 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 99%.

[0437] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.76 (d, J=7.5 Hz, OH), 7.60-7.53 (m, OH), 7.39 (t, J=7.3 Hz, OH), 7.31 (t, J=7.6 Hz, OH), 7.08 (d, J=8.0 Hz, OH), 6.97 (d, J=8.1 Hz, OH), 6.88 (dd, J=15.1, 8.4 Hz, OH), 6.44 (s, OH), 5.43 (s, OH), 4.64 (q, J=6.5 Hz, OH), 4.48-4.37 (m, OH), 4.30 (t, J=8.9 Hz, OH), 4.19 (t, J=6.9 Hz, OH), 3.08-2.94 (m, OH), 1.35 (s, 1H), 1.30 (s, 1H), 1.29 (s, 1H).

[0438] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 170.1, 170.0, 155.8, 154.4, 154.2, 143.7, 143.7, 141.2, 130.8, 130.7, 129.8, 129.8, 127.6, 127.0, 125.0, 124.1, 124.0, 119.9, 82.3, 78.2, 67.1, 56.0, 53.8, 47.0, 37.7, 37.5, 28.8, 28.7, 27.8 ppm.

[0439] HRMS m/z (ESI) calcd for C.sub.45H.sub.55N.sub.2O.sub.7 (M+H).sup.+: 735.4004, found: 735.4000.

Example A17

[0440] ##STR00108##

[0441] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 22) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide (DMF), followed by H-L-Thr(tBu)-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 30 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 90%.

[0442] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.75 (d, J=7.5 Hz, 2H), 7.59 (d, J=7.5 Hz, 2H), 7.39 (t, J=7.5 Hz, 2H), 7.30 (t, J=7.4 Hz, 2H), 6.47 (d, J=8.9 Hz, 1H), 5.61 (d, J=8.1 Hz, 1H), 4.77 (s, 1H), 4.37 (d, J=7.2 Hz, 2H), 4.33 (dd, J=9.0, 2.0 Hz, 1H), 4.27 (q, J=7.3 Hz, 1H), 4.24-4.18 (m, 2H), 3.12 (s, 2H), 2.03 (s, 1H), 1.92-1.84 (m, 1H), 1.76-1.65 (m, 1H), 1.57-1.47 (m, 3H), 1.45 (s, 9H), 1.42 (s, 9H), 1.36-1.29 (m, 1H), 1.25 (t, J=7.1 Hz, 1H), 1.15 (s, 9H).

[0443] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.7, 169.4, 156.0, 143.9, 143.7, 141.2, 127.6, 127.0, 125.1, 119.9, 119.9, 82.0, 73.9, 67.0, 67.0, 58.4, 54.6, 47.1, 32.9, 29.6, 28.7, 28.4, 28.1, 22.2, 21.0 ppm.

[0444] HRMS m/z (ESI) calcd for C.sub.38H.sub.56N.sub.3O.sub.8 (M+H).sup.+: 682.4062, found: 682.4067.

Example A18

[0445] ##STR00109##

[0446] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 28) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide (DMF), followed by H-LVal-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 15 min, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained by column chromatography as a white solid with a yield of 98%.

[0447] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.73 (d, J=7.5 Hz, 2H), 7.57 (d, J=7.5 Hz, 2H), 7.36 (t, J=7.5 Hz, 2H), 7.30-7.16 (m, 17H), 7.04 (s, 1H), 5.96 (d, J=7.2 Hz, 1H), 4.37-4.32 (m, 2H), 4.28 (dd, J=8.3, 4.8 Hz, 1H), 4.19 (t, J=7.5 Hz, 2H), 2.53 (s, 2H), 2.18-2.04 (m, 2H), 2.04-1.91 (m, 2H), 1.41 (s, 9H), 0.84 (d, J=6.8 Hz, 3H), 0.78 (d, J=6.9 Hz, 3H).

[0448] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.8, 171.3, 170.6, 156.1, 144.5, 143.8, 143.7, 141.2, 141.1, 128.6, 127.8, 127.6, 127.0, 126.9, 125.1, 119.8, 81.7, 70.6, 66.9, 58.0, 53.7, 47.0, 33.4, 30.5, 29.8, 27.9, 18.9, 17.5 ppm.

[0449] HRMS m/z (ESI) calcd for C.sub.48H.sub.51N.sub.3NaO.sub.6 (M+Na).sup.+: 788.3670, found: 788.3671.

Example A19

[0450] ##STR00110##

[0451] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 29) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide (DMF), followed by H-L-Leu-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 10 min, and the completion of the reaction was monitored by TLC; after the reaction was completed, the pure product was obtained by column chromatography as a white solid with a yield of 85%.

[0452] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.36 (d, J=68.9 Hz, 2H), 7.43-7.32 (m, 9H), 7.30-7.25 (m, 1H), 6.61 (d, J=8.4 Hz, 1H), 5.51 (d, J=8.7 Hz, 1H), 5.24 (s, 1H), 5.16 (q, J=12.7 Hz, 2H), 4.42 (td, J=8.7, 5.5 Hz, 2H), 4.20 (d, J=7.8 Hz, 1H), 4.08-3.89 (m, 1H), 1.81-1.52 (m, 5H), 1.42 (s, 18H), 1.36-1.27 (m, 2H), 0.86 (t, J=6.1 Hz, 6H).

[0453] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.5, 163.6, 160.6, 155.7, 136.7, 134.6, 128.7, 128.7, 128.4, 128.2, 127.7, 127.7, 81.5, 79.7, 68.8, 66.9, 53.9, 51.2, 44.0, 41.4, 28.4, 28.2, 27.9, 24.8, 24.6, 22.6, 21.9 ppm.

[0454] HRMS m/z (ESI) calcd for C.sub.37H.sub.54N.sub.5O.sub.9 (M+H).sup.+: 712.3916, found: 712.3921.

Example A20

[0455] ##STR00111##

[0456] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 21) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide (DMF), followed by H-L-Thr(tBu)-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 15 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 99%.

[0457] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.75 (d, J=7.5 Hz, 2H), 7.59 (d, J=7.4 Hz, 2H), 7.39 (t, J=7.4 Hz, 2H), 7.30 (t, J=7.4 Hz, 2H), 6.74 (d, J=8.8 Hz, 1H), 5.72 (d, J=8.1 Hz, 1H), 4.52 (q, J=7.2 Hz, 1H), 4.42-4.36 (m, 2H), 4.33 (dd, J=8.9, 1.9 Hz, 1H), 4.26-4.18 (m, 2H), 2.76-2.62 (m, 2H), 2.17-2.00 (m, 5H), 1.46 (s, 9H), 1.16 (s, 9H).

[0458] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.1, 169.3, 155.7, 143.8, 143.7, 141.2, 127.6, 127.0, 125.0, 125.0, 119.9, 119.9, 82.0, 73.8, 66.9, 66.9, 58.6, 53.3, 47.1, 32.2, 29.7, 28.6, 28.0, 21.1, 14.8 ppm.

[0459] HRMS m/z (ESI) calcd for C.sub.32H.sub.45N.sub.2O.sub.6S (M+H).sup.+: 585.2993, found: 585.2991.

Example A21

[0460] ##STR00112##

[0461] In a clean 4 mL reaction vial were added α-carbonyl alkenyl ester (from Preparative Example 30) (0.20 mmol) and 1.5 mL of N,N-dimethylformamide (DMF), followed by H-L-Tyr(tBu)-OtBu (0.22 mmol); then the reaction was stirred at room temperature for 20 min, and the completion of the reaction was monitored by TLC; after the reaction, the pure product was obtained by column chromatography as a white solid with a yield of 99%.

[0462] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.38-7.27 (m, 5H), 7.06 (d, J=8.1 Hz, 2H), 6.91-6.87 (m, 2H), 6.82 (d, J=6.6 Hz, 1H), 5.27 (d, J=8.3 Hz, 1H), 5.14-5.06 (m, 2H), 4.68 (q, J=6.5 Hz, 1H), 4.43-4.36 (m, 1H), 4.28-4.19 (m, 1H), 3.81 (dd, J=8.8, 3.7 Hz, 1H), 3.32 (t, J=8.4 Hz, 1H), 3.07-2.96 (m, 2H), 1.73-1.59 (m, 2H), 1.57-1.48 (m, 1H), 1.35 (s, 9H), 1.31 (s, 9H), 1.17 (s, 9H), 0.92 (t, J=6.2 Hz, 6H).

[0463] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 172.1, 170.0, 169.5, 156.0, 154.3, 136.2, 131.0, 129.9, 128.5, 128.1, 128.0, 123.9, 82.0, 78.2, 74.3, 67.0, 61.1, 53.9, 53.5, 52.9, 41.8, 37.5, 28.8, 27.9, 27.3, 24.7, 22.9, 21.9 ppm.

[0464] HRMS m/z (ESI) calcd for C.sub.38H.sub.58N.sub.3O.sub.8 (M+H).sup.+: 684.4218, found 684.4220.

Application Example B

[0465] A one-pot two-step method was used to prepare amides with the general formula (V).

Example B1

[0466] ##STR00113##

[0467] In a clean 100 mL round-bottom flask were added allenone (10 mmol) and 40 mL of 1,2-dichloroethane (DCE), followed by Fmoc-L-Phe-OH (11 mmol).); then the reaction was stirred at room temperature for 10 h, and the completion of the reaction was monitored by TLC; after the reaction was completed, the reaction solvent was removed in vacuo; thereto 20 mL of N,N-dimethylformamide (DMF) and H-L-Leu-OtBu (11 mmol) were added and stirred at room temperature for 10 min, and the completion of the reaction was monitored by TLC; after the reaction was completed, 100 mL of water was added to the reaction system, and 100 mL of ethyl acetate was used to extract three times, the organic layers combined, the organic layer washed 3 times with 100 mL of water, dried over anhydrous magnesium sulfate and recrystallized to obtain a pure product as a white solid with a yield of 82%.

[0468] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.77 (d, J=7.5 Hz, 2H), 7.55 (t, J=7.8 Hz, 2H), 7.40 (t, J=7.5 Hz, 2H), 7.35-7.15 (m, 7H), 6.59 (s, 1H), 5.63 (d, J=7.1 Hz, 1H), 4.57 (d, J=5.7 Hz, 1H), 4.52-4.41 (m, 2H), 4.33-4.23 (m, 1H), 4.18 (t, J=7.0 Hz, 1H), 3.11 (d, J=5.3 Hz, 2H), 1.68-1.50 (m, 3H), 1.47 (s, 9H), 0.90 (t, J=5.1 Hz, 6H).

[0469] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.5, 170.4, 155.8, 143.70, 143.6, 141.2, 136.2, 129.3, 128.5, 127.6, 127.0, 126.9, 125.0, 124.9, 119.8, 81.8, 67.0, 55.8, 51.4, 47.0, 41.7, 38.5, 27.9, 24.7, 22.6, 22.0 ppm.

[0470] HRMS m/z (ESI) calcd for C.sub.34H.sub.40N.sub.2NaO.sub.5 (M+Na).sup.+: 579.2829, found: 579.2830.

Example B2

[0471] ##STR00114##

[0472] In a clean 100 mL round bottom flask were added allenone (10 mmol) and 40 mL of 1,2-dichloroethane (DCE) followed by Fmoc-L-Ser(tBu)-OH (11 mmol); then the reaction was stirred at room temperature for 5 h, and the completion of the reaction was monitored by TLC; after the reaction was completed, the reaction solvent was removed in vacuo; thereto 20 mL of N,N-dimethylformamide (DMF) and H-L-Leu-OtBu (11 mmol) were then added, the reaction was stirred at room temperature for 10 min, and the completion of the reaction was monitored by TLC; 100 mL of water was added to the reaction system after the reaction, and 100 mL of ethyl acetate was used to extract 3 times, the organic layers combined, the organic layer washed 3 times with 100 mL of water, dried over anhydrous magnesium sulfate, and recrystallized to obtain the pure product as a white solid with a yield of 86%.

[0473] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.75 (d, J=7.5 Hz, 2H), 7.65-7.54 (m, 2H), 7.39 (t, J=7.5 Hz, 2H), 7.31 (t, J=7.5 Hz, 2H), 7.23 (d, 1H), 5.82-5.76 (m, 1H), 4.48 (q, J=7.4 Hz, 1H), 4.43-4.35 (m, 2H), 4.30-4.16 (m, 2H), 3.83 (dd, J=8.5, 3.8 Hz, 1H), 3.40 (t, J=8.4 Hz, 1H), 1.73-1.59 (m, 2H), 1.57-1.51 (m, 1H), 1.46 (s, 9H), 1.22 (s, 9H), 0.95 (d, J=6.4 Hz, 6H).

[0474] .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.6, 169.9, 156.0, 143.9, 143.8, 141.3, 127.7, 127.1, 125.2, 120.0, 81.7, 74.3, 67.1, 61.8, 54.3, 51.7, 47.2, 41.9, 28.0, 27.4, 24.9, 22.8, 22.2 ppm.

[0475] HRMS m/z (ESI) calcd for C.sub.32H.sub.44N.sub.2NaO.sub.6 (M+Na).sup.+: 575.3092, found: 575.3090.

Application Example C

[0476] A solid-phase synthesis method of leucine enkephalin comprising:

[0477] 1) using 2-CTC resin as a carrier, adding Fmoc-Leu-OH and coupling in a solvent in the presence of a base to obtain Fmoc-Leu-resin;

[0478] 2) removing the Fmoc protecting group on the Fmoc-Leu-resin by using a DMF solution containing piperidine to obtain the H-Leu-resin;

[0479] 3) in the presence of a catalyst, adding the α-carbonyl alkenyl ester compound corresponding to Fmoc-Phe-OH, reacting in a solvent (the end of reaction being monitored by Kaiser color test), and washing with a solvent after the reaction is completed (sequentially washing with DCM and DMF) and draining to obtain Fmoc-Phe-Leu-resin;

[0480] 4) repeating the two-step reaction of 2) and 3), and sequentially linking amino acid residue into the peptide chain using α-carbonyl alkenyl esters corresponding to amino acids such as Gly and Tyr according to the amino acid sequence;

[0481] 5) deprotecting, cleaving, and removing the side chain protecting groups and resin, followed by precipitation in ether to obtain crude peptide H-Tyr-Gly-Gly-Phe-Leu-OH (SEQ ID NO:1);

[0482] 6) purifying and freeze-drying to obtain the target polypeptide chain.

[0483] therein, the structure of the H-Tyr-Gly-Gly-Phe-Leu-OH (SEQ ID NO:1) sequence is:

##STR00115##

[0484] wherein, the way of coupling the first amino acid with the 2-CTC resin is to add the first Fmoc-Protected amino acid to a solid-phase reaction vessel, and to link the protected amino acid to the 2-CTC resin under a base reaction condition in a solvent.

[0485] The following examples are only typical examples, which will help to further illustrate and understand the present invention, but these typical examples do not limit the content of the present invention.

Example C1

[0486] 1) Synthesis of Fmoc-Leu-Resin:

[0487] 64.9 mg of 2-CTC resin (loading capacity 0.77 mmol/g) was added to a solid-phase synthesis tube, and 3 mL of dichloromethane was added to soak for 20 min. After soaking, the excess solvent was removed by suction filtration, thereto 0.15 mmol Fmoc-Leu-OH, 0.15 mmol N,N-diisopropylethylamine (DIEA), 1.5 mL DMF and 1.5 mL DCM were added, and the reaction was stirred for 2 h. The reaction solution was filtered off with suction, washed with solvent (DMF), and dried by suction to obtain Fmoc-Leu-resin.

[0488] 2) Synthesis of H-Leu-Resin:

[0489] 0.75 mL of 20% DMF solution of piperidine was added to the solid-phase synthesis tube containing Fmoc-Leu-resin, and the reaction was stirred for 20 min. The reaction solution was filtered off with suction, washed with solvent (DMF) and dried by suction to obtain H-Leu-resin.

[0490] 3) Synthesis of Fmoc-Phe-Leu-Resin:

[0491] To a solid-phase synthesis tube containing H-Leu-resin were add 0.015 mmol HOBt, 0.15 mmol α-carbonyl alkenyl ester corresponding to Fmoc-Phe-OH and 3 mL DMF, and the reaction was stirred (the end of reaction being monitored by Kaiser color test), the reaction solution was filtered off with suction, washed with solvent (DMF), and dried by suction to obtain Fmoc-Phe-Leu-resin.

[0492] 4) Synthesis of H-Tyr(tBu)-Gly-Gly-Phe-Leu-Resin (SEQ ID NO:1 Wherein the First Amino Acid Tyr is Tyr(tBu)):

[0493] Steps 2) and 3) were repeated according to the sequence of H-Tyr(tBu)-Gly-Gly-Phe-Leu-resin (SEQ ID NO:1 wherein the first amino acid Tyr is Tyr(tBu)), the respective α-carbonyl alkenyl esters corresponding to Tyr, Gly, etc. in the sequence were sequentially added to obtain H-Tyr(tBu)-Gly-Gly-Phe-Leu-resin (SEQ ID NO:1 wherein the first amino acid Tyr is Tyr(tBu)).

[0494] 5) The Leucine Enkephalin Resin was Cleaved to Obtain Crude Enkephalin:

[0495] A cleavage cocktail (TFA:TIS:H.sub.2O=95:2.5:2.5) was added to the solid-phase synthesis tube containing H-Tyr(tBu)-Gly-Gly-Phe-Leu-resin (SEQ ID NO:1 wherein the first amino acid Tyr is Tyr(tBu)), reacted at room temperature for 2 h, the solution was filtered out, the resin was washed three times with TFA and filtered, the filtrates were combined, thereto 10 mL of ether was added, the crude product was precipitated, washed with 3×10 mL of ether, and dried to obtain the crude leucine enkephalin product.

[0496] 6) Purification of Crude Leucine Enkephalin:

[0497] 2 mg of the crude leucine enkephalin product was dissolved in 0.25 mL of water, and filtered through a membrane to obtain an aqueous solution of the crude product for later use. C18 reverse chromatographic column was used for separation, mobile phase A: 0.1% TFA, 10% H.sub.2O, 90% MeCN, mobile phase B: 0.1% TFA, 100% H.sub.2O, gradient elution to obtain pure leucine enkephalin solution. The solution was concentrated to 2 mL and lyophilized to obtain pure leucine enkephalin.

[0498] The obtained pure leucine enkephalin was detected by high performance liquid chromatography, and the results are shown in FIG. 1. The obtained pure leucine enkephalin was detected by liquid chromatography-mass spectrometry, and the results are shown in FIG. 2.

Application Example D

[0499] A solid-phase synthesis method for a linear skeleton of oxytocin, comprising:

[0500] 1) using MBHA resin as the carrier, adding α-carbonyl alkenyl ester corresponding to Fmoc-Gly-OH protected by N-terminal Fmoc group, and coupling in solvent in the presence of catalyst to obtain Fmoc-Gly-resin;

[0501] 2) removing the Fmoc protecting group on the Fmoc-Gly-resin by using a DMF solution containing piperidine to obtain the H-Gly-resin;

[0502] 3) in the presence of a catalyst, adding the α-carbonyl alkenyl ester corresponding to Fmoc-Leu-OH and carrying out reaction in a solvent (the end of reaction being monitored by Kaiser color test), washing the resulting resin with a solvent after the reaction is completed (sequentially washed with DCM and DMF) and draining to obtain Fmoc-Leu-Gly-resin;

[0503] 4) repeating the two-step reaction of 2) and 3), and sequentially linking amino acid residue into the peptide chain using α-carbonyl alkenyl esters corresponding to amino acids such as Cys(Me), Tyr, Ile, Gln, Asn, Cys(Me), and Pro according to the amino acid sequence;

[0504] 5) deprotecting, cleaving, and removing the side chain protecting group and resin, and then precipitating in ether to obtain the crude peptide H-Cys(Me)-Tyr-Ile-Gln-Asn-Cys(Me)-Pro-Leu-Gly-OH (SEQ ID NO:2 wherein the first and sixth amino acids Cys are Cys(Me), respectively);

[0505] 6) Purifying and freeze-drying to obtain the target polypeptide chain, wherein, the structure of H-Cys(Me)-Tyr-Ile-Gln-Asn-Cys(Me)-Pro-Leu-Gly-NH.sub.2 sequence (SEQ ID NO:2 wherein the first and sixth amino acids Cys are Cys(Me), respectively) is:

##STR00116##

[0506] wherein, the way of coupling the first amino acid with the MBHA resin is to add the α-carbonyl alkenyl ester corresponding to the first amino acid in a solid-phase reaction vessel, and then link the protected amino acid residue to the MBHA resin in the presence of a catalyst.

Example D1

[0507] 1) Synthesis of Fmoc-Gly-Resin:

[0508] 39.0 mg MBHA resin (loading capacity 0.77 mmol/g) was added to a solid-phase synthesis tube, and 3 mL of dichloromethane was added to soak for 20 min. After soaking, the excess solvent was filtered off with suction, thereto 0.15 mmol of α-carbonyl alkenyl ester corresponding to Fmoc-Gly-OH, 0.015 mmol HOBt and 3 mL DMF were added, and the reaction was stirred to complete (the end of reaction being monitored by Kaiser color test), the reaction solution was filtered off with suction, washed with solvent (DMF), and dried by suction to obtain Fmoc-Gly-resin.

[0509] 2) Synthesis of H-Gly-Resin:

[0510] 0.75 mL of 20% piperidine in DMF solution was added to the solid-phase synthesis tube containing Fmoc-Leu-resin, and the reaction was stirred for 20 min. The reaction solution was filtered off with suction, washed with solvent (DMF), and dried by suction to obtain H-Gly-resin.

[0511] 3) Synthesis of Fmoc-Leu-Gly-Resin:

[0512] In a solid-phase synthesis tube containing H-Gly-resin were added 0.015 mmol HOBt, 0.15 mmol α-carbonyl alkenyl ester corresponding to Fmoc-Leu-OH and 3 mL DMF, and the reaction was stirred to complete (the end of reaction being monitored by Kaiser color test), the reaction solution was filtered off with suction, washed with solvent (DMF), and dried by suction to obtain Fmoc-Leu-Gly-resin.

[0513] 4) Synthesis of H-Cys(Me)-Tyr-Ile-Gln-Asn-Cys(Me)-Pro-Leu-Gly-Resin (SEQ ID NO:2 Wherein the First and Sixth Amino Acids Cys are Cys(Me), Respectively):

[0514] Steps 2) and 3) were repeated according to the sequence of H-Cys(Me)-Tyr-Ile-Gln-Asn-Cys(Me)-Pro-Leu-Gly-resin (SEQ ID NO:2), α-carbonyl alkenyl esters corresponding to amino acids such as Cys(Me), Tyr(tBu), Ile, Gln(Trt), Asn(Trt), Pro were linked sequentially added to obtain H-Cys(Me)-Tyr-Ile-Gln-Asn-Cys(Me)-Pro-Leu-Gly-resin (SEQ ID NO:2 wherein the first and sixth amino acids Cys are Cys(Me), respectively).

[0515] 5) The Oxytocin Linear Skeleton Resin is Cleaved to Obtain the Crude Oxytocin Linear Skeleton:

[0516] The cleavage cocktail (TFA:EDT:TIS:HO=92.5:2.5:2.5:2.5) was added to the solid-phase synthesis tube containing H-Cys(Me)-Tyr-Ile-Gln-Asn-Cys(Me)-Pro-Leu-Gly-resin (SEQ ID NO:2 wherein the first and sixth amino acids Cys are Cys(Me), respectively) and reacted at room temperature for 2 h, the solution was filtered out, the resin washed with TFA for 3 times and filtered, the filtrates were combined, thereto 10 mL of ether was added, and the crude product was separated out, washed with 3×10 mL of ether and dried to obtain crude oxytocin linear peptide.

[0517] 6) Purification of Crude Oxytocin Linear Skeleton:

[0518] 2 mg of the crude oxytocin linear skeleton was dissolved in 0.25 mL of water, and filtered through a membrane to obtain an aqueous solution of the crude peptide for later use. C18 reverse chromatographic column was used for separation, mobile phase A: 0.1% TFA, 10% H.sub.2O, 90% MeCN, mobile phase B: 0.1% TFA, 100% H.sub.2O, gradient elution to obtain pure oxytocin linear skeleton solution. The solution was concentrated to 2 mL and lyophilized to obtain pure oxytocin linear peptide.

[0519] The obtained pure oxytocin linear peptide was detected by high performance liquid chromatography, and the results are shown in FIG. 3. The obtained pure oxytocin linear peptide was tested by liquid chromatography-mass spectrometry, and the results are shown in FIG. 4.

[0520] The above-described examples only describe the preferred embodiment of the invention, and do not limit the scope of the invention. Without departing from the concept and spirit of the invention, various modifications and improvements made by those skilled in the art to the technical scheme of the invention should fall within the protection scope determined by the claims of the invention.

REFERENCES TO THE SEQUENCE LISTING

[0521] Applicant hereby makes reference to the sequence listing that is submitted in electronic format. The Sequence Listing is provided as a file entitled 53208_SEQLIST.txt, created on Jan. 16, 2022 which is 681 bytes in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.