Inhibition of HIV entry is a promising strategy that has not yet been optimally exploited by current drugs. This proposal seeks to develop and characterize a novel class of protease-resistant D-peptides that bind to the highly conserved pocket region of HIV gp41 and block viral entry. D-peptides, composed of mirror-image D- amino acids, are not degraded by natural proteases. Therefore, they have the potential to persist in the body for extended periods of time compared to traditional L-peptide inhibitors, enabling dramatically lower and less frequent dosing at a much lower cost. PIE12-trimer is a highly potent D-peptide inhibitor that broadly inhibits all major HIV clades. It was designed with novel """"""""resistance capacitor"""""""", a reserve of binding energy that is predicted to provide a high genetic barrier to resistance. Indeed, the emergence of resistant strains is much slower for PIE12-trimer than earlier generation D-peptides or the approved entry inhibitor Fuzeon. This proposal will build on the promise of D-peptides and PIE12-trimer by designing and characterizing even more potent membrane-localized variants with pM potency. The properties of these D-peptides in the body will be examined to understand their immunogenicity and metabolic fate. The mechanism by which HIV ultimately resists PIE12-trimer and earlier generation D-peptides will be characterized using deep sequencing to help predict its clinical utility and inform the design of next-generation inhibitors. Novel high-throughput peptide screening techniques will be used to develop next-generation D-peptides that tolerate PIE12-trimer resistance mutations. These studies will advance D-peptide entry inhibitors for use as HIV preventative and therapeutic agents. Understanding how HIV resists this novel class of inhibitors will also improve our understanding of resistance mechanisms and guide the design of inhibitors with more robust resistance profiles. These studies will also validate a rapid modular D-peptide design strategy that can be applied more broadly to inhibit protein- protein interactions for diverse biomedical applications, particularly emerging infectious diseases.
The goal of this project is to optimize novel protease-resistant D-peptide HIV entry inhibitors for use as preventative and therapeutic agents. Detailed studies of how HIV develops resistance to these drugs will inform the design of next-generation entry inhibitors with improved defenses against drug resistance.
|Jacobsen, Michael T; Erickson, Patrick W; Kay, Michael S (2017) Aligator: A computational tool for optimizing total chemical synthesis of large proteins. Bioorg Med Chem 25:4946-4952|
|Jacobsen, Michael T; Petersen, Mark E; Ye, Xiang et al. (2016) A Helping Hand to Overcome Solubility Challenges in Chemical Protein Synthesis. J Am Chem Soc 138:11775-82|
|Petersen, Mark E; Jacobsen, Michael T; Kay, Michael S (2016) Synthesis of tumor necrosis factor ? for use as a mirror-image phage display target. Org Biomol Chem 14:5298-303|
|Weinstock, Matthew T; Jacobsen, Michael T; Kay, Michael S (2014) Synthesis and folding of a mirror-image enzyme reveals ambidextrous chaperone activity. Proc Natl Acad Sci U S A 111:11679-84|
|Weinstock, Matthew T; Francis, J Nicholas; Redman, Joseph S et al. (2012) Protease-resistant peptide design-empowering nature's fragile warriors against HIV. Biopolymers 98:431-42|
|Francis, J Nicholas; Redman, Joseph S; Eckert, Debra M et al. (2012) Design of a modular tetrameric scaffold for the synthesis of membrane-localized D-peptide inhibitors of HIV-1 entry. Bioconjug Chem 23:1252-8|
|Denton, Paul W; Othieno, Florence; Martinez-Torres, Francisco et al. (2011) One percent tenofovir applied topically to humanized BLT mice and used according to the CAPRISA 004 experimental design demonstrates partial protection from vaginal HIV infection, validating the BLT model for evaluation of new microbicide candidates. J Virol 85:7582-93|
|Welch, Brett D; Francis, J Nicholas; Redman, Joseph S et al. (2010) Design of a potent D-peptide HIV-1 entry inhibitor with a strong barrier to resistance. J Virol 84:11235-44|
|Eckert, Debra M; Kay, Michael S (2010) Stalking influenza. Proc Natl Acad Sci U S A 107:13563-4|
|Eckert, Debra M; Shi, Yu; Kim, Sunghwan et al. (2008) Characterization of the steric defense of the HIV-1 gp41 N-trimer region. Protein Sci 17:2091-100|