In vitro activities do not always translate into in vivo activities, and this is true in Mycobacterium tuberculosis (Mtb) therapeutics. For example, the two first line anti-Mtb drugs isoniazid and rifampicin kill Mtb rapidly in vitro, but their steriliation abilities are slowed and reduced in vivo. Heterogeneity of Mtb populations imparts varied susceptibility to the drugs. In particular, slowly-replicating or non-replicating (collectively """"""""NR) Mtb is non-heritably resistant to most first line anti-TB drugs. Our long-term goal is to develop anti-Mtb drugs that kill NR Mtb populations to complement drugs that kill replicating Mtb populations. During the past eight years, components of a prokaryotic ubiquitin-like protein (Pup)-proteasome system have been discovered. Although the Mtb proteasome is dispensable under standard growth conditions, genetic evidence demonstrates its essentiality for Mtb to survive in mice. We also established the concept that despite the essential role of the proteasome in mammals, small chemical molecules can be discovered with extensive (>1000-fold) species selectivity for inhibiting the Mtb proteasome over the human proteasome, and that such inhibition leads to killing of NR Mtb. In other bacteria, disruption of another regulated protein degradation machine, Clp, has led to killing. Thus, either inhibition or forced activation f chambered proteases may represent a new anti-infective strategy. In our recent screening of 1600 capped dipeptides, we identified a lead compound that potently and species-selectively inhibited the Mtb proteasome over the human proteasome. The inhibitors were bactericidal for non-replicating Mtb in synergy with nitric oxide. Using a competition assay, I confirmed that the dipeptides penetrate the mycobacteria and inhibit the proteasome within them. In this application, we will expand our lead compounds into a small focused compound library whose design and synthesis are guided by substrate-profiling, structural analysis, mycobactericidal activity, and metabolic stability.

Public Health Relevance

Mycobacterium tuberculosis (Mtb) infection worldwide causes 1.5 million deaths with 8-9 million new cases annually. Our goal is to develop small chemical molecules that target a protein-degrading enzyme of Mtb that plays pivotal roles in its survival in the host. The study will provide leads compounds for further development as anti-TB therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI101393-01A1
Application #
8510791
Study Section
Drug Discovery and Mechanisms of Antimicrobial Resistance Study Section (DDR)
Program Officer
Lacourciere, Karen A
Project Start
2013-02-01
Project End
2015-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
1
Fiscal Year
2013
Total Cost
$253,500
Indirect Cost
$103,500
Name
Weill Medical College of Cornell University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
060217502
City
New York
State
NY
Country
United States
Zip Code
10065
Kirkman, Laura A; Zhan, Wenhu; Visone, Joseph et al. (2018) Antimalarial proteasome inhibitor reveals collateral sensitivity from intersubunit interactions and fitness cost of resistance. Proc Natl Acad Sci U S A 115:E6863-E6870
Totaro, Kyle A; Barthelme, Dominik; Simpson, Peter T et al. (2017) Rational Design of Selective and Bioactive Inhibitors of the Mycobacterium tuberculosis Proteasome. ACS Infect Dis 3:176-181
Hsu, Hao-Chi; Singh, Pradeep K; Fan, Hao et al. (2017) Structural Basis for the Species-Selective Binding of N,C-Capped Dipeptides to the Mycobacterium tuberculosis Proteasome. Biochemistry 56:324-333
Santos, Ruda de Luna Almeida; Bai, Lin; Singh, Pradeep K et al. (2017) Structure of human immunoproteasome with a reversible and noncompetitive inhibitor that selectively inhibits activated lymphocytes. Nat Commun 8:1692
Singh, Pradeep K; Fan, Hao; Jiang, Xiuju et al. (2016) Immunoproteasome ?5i-Selective Dipeptidomimetic Inhibitors. ChemMedChem 11:2127-2131
Lin, Gang; Chidawanyika, Tamutenda; Tsu, Christopher et al. (2013) N,C-Capped dipeptides with selectivity for mycobacterial proteasome over human proteasomes: role of S3 and S1 binding pockets. J Am Chem Soc 135:9968-71