| 摘要 |
<p>METHOD FOR PREPARING PEPTIDES BY ASSEMBLING MULTIPLE PEPTIDE FRAGMENTS5Method for preparing a peptide assembly of n fragments and n-1 amino acids bearing a thiol function, represented by the formula:10 A1-C1-A2-C2-A3- -C1-An (I)in which A1, A2, A3, A, ..., An are peptide fragments, Ci , C2, C3 CI-1 Cn-i are aminoacid residues bearing a thiol function, n is comprised between 3 and 50, and i is 2 to n, in which a peptide-thioester is prepared of formula: Al-SR (II) in which A1 is a peptide fragment and SR is an alkyl thioester residue, R being alkyl optionally substituted, starting from a15 bis(2-sulphanylethyl)amino peptide: Al-SEAoff in which SEAoff is a cyclic bis(2- sulphanylethyl)amino group, by action of a thiol R-SH, in the presence of a cyclic disulphide reducing agent, followed by condensation with a peptide fragment of structure: H-C1(SR')-A2- SEAoff (III) in which (SR') represents a disulphide residue on the thiol of the amino acid C1, in the presence of an aromatic thiol ArSH, followed by conversion of the obtained new20 peptide, of structure:A1-C1-A2-SEAoff (IV), to a peptide-thioester of formula: A1-C1-A2-SR (II'), by action of a thiol R-SH, in the presence of a cyclic disulphide reducing agent, and condensation with a peptide fragment of structure H-C2(SIT)-A3-SEAoff (III') in the presence of an aromatic thiol ArSH, in order to produce a peptide fragment of structure A1-C1-A2-C2-A3- SEAoff (IV') and reiteration of these 2 operations up to n-2 times, in order to obtain a peptide25 fragment of structure ArC1-A2-C2-A3---C1_1-A,----C,2-An_1-SEAoff (IVm), then implementation of a native ligation reaction of this obtained peptide fragment (IVm), with a peptide of formula: H-C1-An (Vn) by reduction in the presence of a cyclic disulphide reducing agent, in order to produce the multiple peptide assembly of general formula (I).30</p> |
