@article {43267, title = {High-throughput screening of RNA polymerase inhibitors using a fluorescent UTP analog}, journal = {Journal of Biomolecular Screening}, volume = {11}, year = {2006}, month = {DEC}, pages = {968-976}, abstract = {

RNA polymerase (RNAP) is a well-validated target for the development of antibacterial and antituberculosis agents. Because the purification of large quantities of native RNA polymerase from pathogenic mycobacteria is hazardous and cumbersome, the primary screening was carried out using Escherichia coli RNAP. The authors have developed a high-throughput screening (HTS) assay to screen for novel inhibitors of RNAP. In this assay, a fluorescent analog of UTP, gamma-amino naphthalene sulfonic acid (gamma-AmNS) UTP, was used as one of the nucleotide substrates. Incorporation of UMP in RNA results in the release of gamma-AmNS-PPi, which has higher intrinsic fluorescence than (gamma-AmNS) UTP. The assay was optimized in a 384-well format and used to screen 670,000 compounds at a concentration of 10 mu M. About 0.1\% of the compounds showed more than 60\% inhibition in the primary HTS. All the primary actives tested for dose response using the same assay had an EC50 below 100 mu M. Eighty percent of the primary HTS actives obtained using E. coli RNAP showed comparable activity against Mycobacterium smegmatis RNAP in the conventional radioactive assay. Activity of hits selected for the hit-to-lead optimization was also confirmed against Mycobacterium bovis RNAP which has \> 99\% sequence identity with Mycobacterium tuberculosis RNAP subunits.

}, author = {Bhat, Jyothi and Rane, Rajendra and Solapure, Suresh M. and Sarkar, Dhiman and Sharma, Umender and Harish, M. N. and Lamb, Sarah and Plant, Darren and Alcock, Peter and Peters, Steve and Barde, Shubhada and Roy, Raman K.} } @article { ISI:000330241000045, title = {Highly efficient one-pot multi-component synthesis of alpha-aminophosphonates and bis-alpha-aminophosphonates catalyzed by heterogeneous reusable silica supported dodecatungstophosphoric acid (DTP/SiO2) at ambient temperature and their antitubercular eval}, journal = {RSC Advances}, volume = {4}, number = {15}, year = {2014}, month = {JAN}, pages = {7666-7672}, publisher = {ROYAL SOC CHEMISTRY}, address = {THOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLAND}, abstract = {

A highly efficient one-pot multi-component reaction (MCR) protocol over DTP/SiO2 has been developed for the synthesis of alpha-aminophosphonate derivatives (4a-x) in excellent yields. The alpha-aminophosphonate derivatives were for the first time evaluated for their antitubercular activity against the M. tuberculosis H37Ra (MTB) strain. An evaluation of the data on the cytotoxicity and antimicrobial activity shows that 4n and 4v are promising antitubercular agents.

}, issn = {2046-2069}, doi = {10.1039/c3ra45853a}, author = {Mulla, Shafeek Abdul Rashid and Pathan, Mohsinkhan Y. and Chavan, Santosh S. and Gample, Suvarna P. and Sarkar, Dhiman} } @conference {42974, title = {High throughput screening of inhibitors for tuberculosis at NCL : a novel drug discovery initiative from CSIR}, booktitle = {73rd CSIR Foundation Day, at CSIR-National Chemical Laboratory, Pune}, year = {2015}, month = {SEP}, publisher = {CSIR-National Chemical Laboratory, Pune}, organization = {CSIR-National Chemical Laboratory, Pune}, address = {CSIR-National Chemical Laboratory, Pune}, abstract = {

Discover and develop novel drugs that are effective at curing latent, drug sensitive and drug resistant TB Study the biology of tuberculosis bacilli in great detail to identify new metabolic pathways that may be vulnerable to drugs Test thousands of potential drugs in our screening facility to find new compounds or new compound classes that kill TB bacteria Increase throughput up to 20,000 compounds at a time Develop novel screens using conditions that mimic those the TB bacteria encounters in the human body Utilize medical chemistry to optimize the structures of compounds so that they kill TB bacteria in a more potent fashion and are non-toxic Meet the challenges from industry

}, author = {Manek, Hardik and Nawale, Laxman and Jadhav, Nandadeep J. and Sarkar, Dhiman} } @article {43264, title = {Hybrids of thienopyrimidinones and thiouracils as anti-tubercular agents: SAR and docking studies}, journal = {European Journal of Medicinal Chemistry}, volume = {127}, year = {2017}, month = {JAN}, abstract = {

A number of hybrid molecules containing thienopyrimidinones and thiouracil moieties were designed, synthesized and tested against Mycobacterium tuberculosis H37Ra wherein it was observed that the compounds 11-1514 exhibited antitubercular activity in vitro (MIC 7.6{\textendash}19.1 μg/ml, 12{\textendash}35 μM) against dormant stage while compound 15 exhibited antitubercular activity in vitro against dormant (MIC 23.4 μg/ml, 41 μM) as well as active (MIC 25.4 μg/ml, 45 μM) stage. Structural modifications of compound 15 were carried out to study the structure-activity relationship and it was observed that compound 18 exhibited antitubercular activity comparable to compound 15. Cytotoxicity studies revealed that these molecules were non-toxic. The docking study of compound 15 showed that there was binding with the active site of mycobacterial pantothenate synthetase. Further docking studies led to the synthesis of the compounds 16 and 17 and the antitubercular activity screening results showed that these compounds have significant antitubercular activity. The compounds 15-18 (MIC 11{\textendash}29 μg/ml, 19{\textendash}51 μM) can be used as starting points for further optimization. The synthetic strategies used in the present work have potential to prepare a large number of compounds for further refinement of structures and the present results will be very useful in the development of a new class of antimycobacterial agents.

}, doi = {10.1016/j.ejmech.2017.01.009}, author = {Pisal, Mahesh M. and Nawale, Laxman U. and Patil, Manoj and Bhansali, Sujit G. and Gajbhiye, Jayant M. and Sarkar, Dhiman and Chavan, Subhash P. and Borate, Hanumant B.} } @article {46284, title = {High content screening assay for identifying inhibitors against active and dormant state intracellular Mycobacterium tuberculosis}, journal = {Journal of Microbiological Methods}, volume = {164}, year = {2019}, month = {SEP}, type = {Article}, abstract = {

The antitubercular drug development pipeline could start with an in vitro investigation of several compounds to examine their effect on active and dormant state Mycobacterium tuberculosis (Mtb). However, in vitro screening of dormant state bacilli cannot provide enough information on the simultaneous effect of a compound on the host. Therefore, we developed a live cell fluorescence based screening protocol by utilizing the high content system for determining the effect of inhibitors against active and dormant state intracellular mycobacteria. THP-1 macrophages infected with an actively growing and hypoxia derived dormant Mtb culture were standardized to develop the screening protocol. The signal to noise ratio and the Z{\textquoteright} factor of this assay were found to be 7.5-29 and 0.6-0.8, respectively, which confirm the robustness of the protocol. The protocol was then validated with standard inhibitors. This newly developed drug screening assay offers an easy, safe, image based high content screening tool to search for novel antitubercular inhibitors against both active and dormant state intracellular mycobacteria. Therefore, this assay could fill in the gap between in vitro and in vivo latent tuberculosis drug screening programs.

}, doi = {10.1016/j.mimet.2019.105687}, author = {Yeware, Amar and Agrawal, Sonia and Sarkar, Dhiman} }