title: A combinatorial approach for the design of complementarity-determining region-derived peptidomimetics with in vitro anti-tumoral activity.
abstract: The great success of therapeutic monoclonal antibodies has fueled research toward mimicry of their binding sites and the development of new strategies for peptide-based mimetics production. Here, we describe a new combinatorial approach for the production of peptidomimetics using the complementarity-determining regions (CDRs) from gastrin17 (pyroEGPWLEEEEEAYGWMDF-NH(2)) antibodies as starting material for cyclic peptide synthesis in a microarray format. Gastrin17 is a trophic factor in gastrointestinal tumors, including pancreatic cancer, which makes it an interesting target for development of therapeutic antibodies. Screening of microarrays containing bicyclic peptidomimetics identified a high number of gastrin binders. A strong correlation was observed between gastrin binding and overall charge of the peptidomimetic. Most of the best gastrin binders proceeded from CDRs containing charged residues. In contrast, CDRs from high affinity antibodies containing mostly neutral residues failed to yield good binders. Our experiments revealed essential differences in the mode of antigen binding between CDR-derived peptidomimetics (K(d) values in micromolar range) and the parental monoclonal antibodies (K(d) values in nanomolar range). However, chemically derived peptidomimetics from gastrin binders were very effective in gastrin neutralization studies using cell-based assays, yielding a neutralizing activity in pancreatic tumoral cell lines comparable with that of gastrin-specific monoclonal antibodies. These data support the use of combinatorial CDR-peptide microarrays as a tool for the development of a new generation of chemically synthesized cyclic peptidomimetics with functional activity.
authors.affiliation: Pepscan Therapeutics B.V., Zuidersluisweg 2, 8243 RC Lelystad, The Netherlands. p.timmerman@pepscan.com
authors.forename: Peter
authors.initials: P
authors.lastname: Timmerman
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Barderas
authors.forename: Johan
authors.initials: J
authors.lastname: Desmet
authors.forename: Danièle
authors.initials: D
authors.lastname: Altschuh
authors.forename: Susana
authors.initials: S
authors.lastname: Shochat
authors.forename: Martine J
authors.initials: MJ
authors.lastname: Hollestelle
authors.forename: Jo W M
authors.initials: JW
authors.lastname: Höppener
authors.forename: Alberto
authors.initials: A
authors.lastname: Monasterio
authors.forename: J Ignacio
authors.initials: JI
authors.lastname: Casal
authors.forename: Rob H
authors.initials: RH
authors.lastname: Meloen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000915:Antibody Affinity; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D020650:Combinatorial Chemistry Techniques; D022801:Complementarity Determining Regions / Q000737:chemistry* / Q000235:genetics*; D005755:Gastrins / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D051379:Mice; D008968:Molecular Conformation; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D022801:Complementarity Determining Regions; D005755:Gastrins; D019151:Peptide Library; D010455:Peptides