Article abstract
Nature Chemical Biology 5, 502 - 507 (2009)
Published online: 31 May 2009 | doi:10.1038/nchembio.184
Phage-encoded combinatorial chemical libraries based on bicyclic peptides
Christian Heinis1,3, Trevor Rutherford2, Stephan Freund2 & Greg Winter1
Abstract
Here we describe a phage strategy for the selection of ligands based on bicyclic or linear peptides attached covalently to an organic core. We designed peptide repertoires with three reactive cysteine residues, each spaced apart by several random amino acid residues, and we fused the repertoires to the phage gene-3-protein. Conjugation with tris-(bromomethyl)benzene via the reactive cysteines generated repertoires of peptide conjugates with two peptide loops anchored to a mesitylene core. Iterative affinity selections yielded several enzyme inhibitors; after further mutagenesis and selection, we were able to chemically synthesize a lead inhibitor (PK15; Ki = 1.5 nM) specific to human plasma kallikrein that efficiently interrupted the intrinsic coagulation pathway in human plasma tested ex vivo. This approach offers a powerful means of generating and selecting bicyclic macrocycles (or if cleaved, linear derivatives thereof) as ligands poised at the interface of small-molecule drugs and biologics.
- Laboratory of Molecular Biology, Medical Research Council, Cambridge, UK.
- Centre for Protein Engineering, Medical Research Council, Cambridge, UK.
- Present address: Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Correspondence to: Christian Heinis1,3 e-mail: christian.heinis@epfl.ch
Correspondence to: Greg Winter1 e-mail: winter@mrc-lmb.cam.ac.uk
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