Merck
  • Home
  • Search Results
  • Assessment of Mycobacterium tuberculosis pantothenate kinase vulnerability through target knockdown and mechanistically diverse inhibitors.

Assessment of Mycobacterium tuberculosis pantothenate kinase vulnerability through target knockdown and mechanistically diverse inhibitors.

Antimicrobial agents and chemotherapy (2014-04-02)
B K Kishore Reddy, Sudhir Landge, Sudha Ravishankar, Vikas Patil, Vikas Shinde, Subramanyam Tantry, Manoj Kale, Anandkumar Raichurkar, Sreenivasaiah Menasinakai, Naina Vinay Mudugal, Anisha Ambady, Anirban Ghosh, Ragadeepthi Tunduguru, Parvinder Kaur, Ragini Singh, Naveen Kumar, Sowmya Bharath, Aishwarya Sundaram, Jyothi Bhat, Vasan K Sambandamurthy, Christofer Björkelid, T Alwyn Jones, Kaveri Das, Balachandra Bandodkar, Krishnan Malolanarasimhan, Kakoli Mukherjee, Vasanthi Ramachandran
ABSTRACT

Pantothenate kinase (PanK) catalyzes the phosphorylation of pantothenate, the first committed and rate-limiting step toward coenzyme A (CoA) biosynthesis. In our earlier reports, we had established that the type I isoform encoded by the coaA gene is an essential pantothenate kinase in Mycobacterium tuberculosis, and this vital information was then exploited to screen large libraries for identification of mechanistically different classes of PanK inhibitors. The present report summarizes the synthesis and expansion efforts to understand the structure-activity relationships leading to the optimization of enzyme inhibition along with antimycobacterial activity. Additionally, we report the progression of two distinct classes of inhibitors, the triazoles, which are ATP competitors, and the biaryl acetic acids, with a mixed mode of inhibition. Cocrystallization studies provided evidence of these inhibitors binding to the enzyme. This was further substantiated with the biaryl acids having MIC against the wild-type M. tuberculosis strain and the subsequent establishment of a target link with an upshift in MIC in a strain overexpressing PanK. On the other hand, the ATP competitors had cellular activity only in a M. tuberculosis knockdown strain with reduced PanK expression levels. Additionally, in vitro and in vivo survival kinetic studies performed with a M. tuberculosis PanK (MtPanK) knockdown strain indicated that the target levels have to be significantly reduced to bring in growth inhibition. The dual approaches employed here thus established the poor vulnerability of PanK in M. tuberculosis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Isopropyl β-D-1-thiogalactopyranoside, ≥99% (TLC)
Supelco
Isoniazid, analytical standard, ≥99% (TLC)
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
Isopropyl β-D-thiogalactopyranoside solution, ReadyMade IPTG solution for Blue-white screening
Supelco
Isoniazid, Vetec, reagent grade, 98%
Isoniazid, European Pharmacopoeia (EP) Reference Standard
SAFC
Isopropyl β-D-1-thiogalactopyranoside
Isopropyl β-D-thiogalactoside, Vetec, reagent grade, ≥99%