Method for the synthesis and selective biocatalytic modification of peptides, peptide mimetics and proteins

The method as claimed in one or more of the preceding claims,

• characterized in that
• the amino component has a size of from 1 to 1000 amino acids, preferably from 30 to 500 amino acids, more preferably from 30 to 250 amino acids.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the carboxyl end of the amino component is present protected or unprotected in nonactivated form.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the Nα-amino function of the amino component is present unprotected, and this Nα-amino function reacts with the carboxyl function of the carboxyl component.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the carboxyl component is a polypeptide or protein.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the carboxyl component has a size of from 1 to 1000 amino acids, preferably from 30 to 500 amino acids, more preferably from 30 to 250 amino acids.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the carboxyl group of the carboxyl component forms a carboxylic ester or a carboxamide with the leaving group.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the leaving group of the carboxyl component is selected from the group consisting of unsubstituted and substituted —O-alkyl-, —O-aryl-, —S-alkyl-, —S-aryl radicals, —NH-alkyl-, —NH-aryl-, —N,N-dialkyl-, —N,N-diaryl- and —N-aryl-N-alkyl-radicals.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the leaving group is a 4-guanidinophenyl, 4-amidinophenyl, 4-guanidinophenylthio or 4-amidinophenylthio radical, or a compound structurally homologous thereto.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the leaving group is adjusted to the specificity of the protease, peptidase and/or hydrolase used.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the steps a) to c) and optionally d) are carried out twice or more, in order to prepare sequentially a polypeptide or protein which may have a label or reporter group.

The method as claimed in one or more of the preceding claims,

• characterized in that
• a compound prepared according to steps a) to c) and optionally d) is used as the amino component and another compound prepared according to steps a) to c) and optionally d) is used as the carboxyl component.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the reaction medium has exclusively one or more ionic liquids.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the proportion of ionic liquids in the reaction medium is 50-100% by volume, preferably 70-100 or 80-100% by volume, more preferably 90-100% by volume, likewise preferably from 95 to 100% by volume or 95-99% by volume.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the cations of the ionic liquids used are quaternized alkylimidazolium ions, quaternized alkylammonium ions, quaternized alkylpyridinium ions and/or quaternized alkylphosphonium ions.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the ionic liquids used are 1-ethyl-3-methylimidazolium, 1-butyl-3-methylimidazolium and/or 4-methyl-N-butylpyridinium salts.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the anions of the ionic liquids used are chloride, bromide, chloroaluminate, nitrate, benzenesulfonate, triflate(trifluoromethanesulfonate), tosylate and/or tetrafluoroborate.

The method as claimed in one or more of the preceding claims,

• characterized in that
• the protease used is a cysteine protease or serine protease.

The use of ionic liquids as an exclusive solvent or in combination with water and/or organic solvents for the synthesis and/or N-terminal modification of peptides, peptide mimetics and/or proteins.

The use of a protease, peptidase and/or hydrolase for the synthesis and/or N-terminal modification of peptides, peptide mimetics and proteins, said peptide, peptide mimetic and protein or N-terminally labeled species thereof being prepared by ligation of an amino component and a carboxyl component, and said carboxyl component having a leaving group.