Directed Evolution Using Stabilized Bacterial Peptide Display
- 10 January 2020
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 142 (4), 1882-1894
- https://doi.org/10.1021/jacs.9b10716
Abstract
Chemically stabilized peptides have attracted intense interest by academics and pharmaceutical companies due to their potential to hit currently ‘undruggable’ targets. However, engineering an optimal sequence, stabilizing linker location, and physicochemical properties is a slow and arduous process. By pairing non-natural amino acid incorporation and cell surface click chemistry in bacteria with high-throughput sorting, we developed a method to quantitatively select high affinity ligands and applied the SPEED (Stabilized Peptide Evolution by E. coli Display) technique to develop disrupters of the therapeutically relevant MDM2-p53 interface. Through in situ stabilization on the bacterial surface, we demonstrate rapid isolation of stabilized peptides with improved affinity and novel structures. Several peptides evolved a second loop including one sequence (Kd = 1.8 nM) containing an i, i+4 disulfide bond. NMR structural determination indicated a bent helix in solution and bound to MDM2. The bicyclic peptide had improved protease stability, and we demonstrated that protease resistance could be measured both on the bacterial surface and in solution, enabling the method to test and/or screen for additional drug-like properties critical for biologically active compounds.Keywords
Funding Information
- National Science Foundation (1553860)
- U.S. Department of Health and Human Services (R35 GM128819)
- National Science Foundation
This publication has 72 references indexed in Scilit:
- Preliminary Results of the Stapled Peptide ALRN-6924, a Dual Inhibitor of MDMX and MDM2, in Two Phase IIa Dose Expansion Cohorts in Relapsed/Refractory TP53 Wild-Type Peripheral T-Cell LymphomaBlood, 2018
- Macrocyclic α helical peptide therapeutic modality: A perspective of learnings and challengesBioorganic & Medicinal Chemistry, 2018
- Iterative optimization yields Mcl-1–targeting stapled peptides with selective cytotoxicity to Mcl-1–dependent cancer cellsProceedings of the National Academy of Sciences of the United States of America, 2018
- Biophysical determinants for cellular uptake of hydrocarbon-stapled peptide helicesNature Chemical Biology, 2016
- A Helix-Stabilizing Linker Improves Subcutaneous Bioavailability of a Helical Peptide Independent of Linker LipophilicityBioconjugate Chemistry, 2016
- Discovery of the Once-Weekly Glucagon-Like Peptide-1 (GLP-1) Analogue SemaglutideJournal of Medicinal Chemistry, 2015
- Stapled α−helical peptide drug development: A potent dual inhibitor of MDM2 and MDMX for p53-dependent cancer therapyProceedings of the National Academy of Sciences of the United States of America, 2013
- Reactivation of the p53 Tumor Suppressor Pathway by a Stapled p53 PeptideJournal of the American Chemical Society, 2007
- Small-molecule inhibitors of protein–protein interactions: progressing towards the dreamNature Reviews Drug Discovery, 2004
- The druggable genomeNature Reviews Drug Discovery, 2002