Iterative in Situ Click Chemistry Assembles a Branched Capture Agent and Allosteric Inhibitor for Akt1

作者：Millward Steven W; Henning Ryan K; Kwong Gabriel A; Pitram Suresh; Agnew Heather D; Deyle Kaycie M; Nag Arundhati; Hein Jason; Lee Su Seong; Lim Jaehong; Pfeilsticker Jessica A; Sharpless K Barry; Heath James R^*

来源：Journal of the American Chemical Society, 2011, 133(45): 18280-18288.

DOI：10.1021/ja2064389

We describe the use of iterative in situ click chemistry to design an Akt-specific branched peptide triligand that is a drop-in replacement for monoclonal antibodies in multiple biochemical assays. Each peptide module in the branched structure makes unique contributions to affinity and/or specificity resulting in a 200 nM affinity ligand that efficiently immunoprecipitates Akt from cancer cell lysates and labels Akt in fixed cells. Our use of a small molecule to preinhibit Akt prior to screening resulted in low micromolar inhibitory potency and an allosteric mode of inhibition, which is evidenced through a series of competitive enzyme kinetic assays. To demonstrate the efficiency and selectivity of the protein-templated in situ click reaction, we developed a novel QPCR-based methodology that enabled a quantitative assessment of its yield. These results point to the potential for iterative in situ click chemistry to generate potent, synthetically accessible antibody replacements with novel inhibitory properties.

• 出版日期2011-11-16