Synthesis of a phosphotyrosyl analogue having X-1, X-2 and phi angles constrained to values observed for an SH2 domain-bound phosphotyrosyl residue

Src homology 2 (SH2) domains provide connectivity in protein- tyrosine kinase (PTK)-dependent signaling through their high affinity association with phosphotyrosyl (pTyr)-containing peptide sequences. Because recognition of pTyr residues is central to SH2 domain-binding affinity, design of pTyr- mimicking residues has been one component of SH2 domain signaling antagonist development. Reported herein is the synthesis of (+/-)-(rel-1R,2R,5S)-3-acetyl-1,2,3,4,5,6- hexahydro-8-O-phosphoryl-1,5-methano-3-benzazocine-2carboxylic acid methyl ester (3c) as a monomeric pTyr-mimicking analogue that constrains three torsion angles (chi(1) = 168degrees; chi(2)= -85degrees; phi(1) = -113degrees) to values approximating those observed for a pTyr residue bound to the Grb2 SH2 domain (chi(1) = 182degrees; chi(2)= -89degrees; phi(1) = -132degrees). Compound 3c differs from our previously reported analogue, (+/-)-(rel-1R,2R,5S)-3-acetyl-1,2,3,4,5,6- hexahydro-1-methyl-1,5-methano-3-benzazocin-8-ol. in lacking a methyl substituent at the bridgehead 1-position. Molecular modeling studies had indicated that this methyl group could potentially hinder SH2 domain binding. Synthesis of the desmethyl derivative was achieved by formation of the methanobenzazocine ring system using an intramolecular electrophilic cyclization that proceeds through an activated acyliminium intermediate. Importantly, the correct relative (2R) stereochemistry at the 'alpha-carboxyl'-bearing carbon is obtained through base-catalyzed equilibration of a (2S/2R) diastereomeric mixture that results from intramolecular ring closure. Comparison of Grb2 SH2 domain-binding affinity of 3c (IC50=1167 muM) with conformationally flexible phosphorylated (+/-)-N-acetyl-tyrosine methyl ester (15; IC50=1469 muM) revealed no apparent enhancement in affinity. This apparent ineffectiveness of 'local conformational constraint' on SH2 domain-binding affinity of the monomeric pTyr mimetic is consistent with previous reports obtained by conformationally constraining pTyr-mimicking residues that were contained within peptide platforms. Although not providing high binding affinity in its current form, the novel 1,5-methano-3-benzazocine ring system may afford a novel platform for further elaboration and development of small molecule SH2 domain signaling antagonists. (C) 2003 Elsevier Ltd. All rights reserved.

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