Method for the poduction of cyclic molecules
AssigneeTransmit Gesellschaft fur Technologietransfer mbH
InventorsMohamed Marahiel, Stephan Sieber
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Abstract
The invention at hand describes a method for the cyclization of peptides and proteins in which linear thioesters serve as substrates. The cyclization is catalyzed by thioesterase domains of NRPS or PKS cyclases. The substrates according to the present invention are composed of one linear peptide on which a charge-stabilized aromatic, heteroaromatic or araliphatic leaving group is bound. These substrates lead to higher yields and reaction rates than linear peptides able to be cyclized with methods known so far and, furthermore, allow the cyclization of such peptides which were previously not able to be cyclized.
Claims16
Method for the production of cyclic peptides, in which
- a peptide cyclase is brought in contact with a linear peptide,
- the linear peptide contains an acyl residue, which is activated by a nucleophilic leaving group bound chemically with this acyl residue,
- the activated acyl residue of the linear peptide selectively acylates the center of the peptide cyclase, wherein the nucleophilic leaving group is cleaved off during formation of the cyclic peptide and
- cyclic peptides with rings of at least 5 atoms are formed,
- wherein
- the nucleophilic leaving group, which is chemically bound to the acyl residue of the linear peptide and which activates the latter, is charge-stabilized and
- the charge-stabilized leaving group is bound to the acyl group of the C-terminal carboxylic acid group.
Method for the production of a substrate and subsequent reaction of this substrate with peptide cyclases into a cyclic peptide, wherein the substrates are linear peptides, wherein the following steps are carried out one after the other:
- Adding a reagent activating the C-terminus of the peptide acid, a coupling additive and a charge-stabilized leaving group to the free peptide acid in a solvent
- Stirring at room temperature,
- Addition of a base and further stirring at room temperature,
- Filtration,
- Removal of the solvent,
- Deprotection of the peptide,
- Addition of a peptide cyclase,
- Purification of the cyclic peptide obtained.
Dependencies are inferred from the claim text.