Identification of Shc Src Homology 2 Domain-Binding Peptoid-Peptide Hybrids

A fluorescence anisotropy (FA) competition-based Shc Src homology 2 (SH2) domain-binding was established using the high affinity fluorescein isothiocyanate (FITC) containing peptide, FITC-NH-(CH2)(4)-CO-pY-Q-G-L-S-amide (8; K-d = 0.35 mu M). Examination of a series of open-chain bis-alkenylamide containing peptides, prepared as ring-closing metathesis precursors, showed that the highest affinities were obtained by replacement of the original Gly residue with N-alpha-substituted Gly (NSG) peptoid residues. This provided peptoid-peptide hybrids of the form Ac-pY-Q-[NSG]-L-amide. Depending on the NSG substituent, certain of these hybrids exhibited up to 40-fold higher Shc SH2 domain-binding affinity than the parent Gly-containing peptide (IC50 = 248 mu M) (for example, for N-homoallyl analogue 50, IC50 = 6 mu M). To our knowledge, this work represents the first successful example of the application of peptoid-peptide hybrids in the design of SH2 domain-binding antagonists. These results could provide a foundation for further structural optimization of She SH2 domain-binding peptide mimetics.