02207nas a2200265 4500008004100000245009000041210006900131260000800200300001200208490000700220520136600227100001701593700001901610700002501629700001901654700002001673700001901693700001601712700001801728700001801746700001801764700002001782700001901802856012001821 2006 eng d00aHigh-throughput screening of RNA polymerase inhibitors using a fluorescent UTP analog0 aHighthroughput screening of RNA polymerase inhibitors using a fl cDEC a968-9760 v113 a

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.

1 aBhat, Jyothi1 aRane, Rajendra1 aSolapure, Suresh, M.1 aSarkar, Dhiman1 aSharma, Umender1 aHarish, M., N.1 aLamb, Sarah1 aPlant, Darren1 aAlcock, Peter1 aPeters, Steve1 aBarde, Shubhada1 aRoy, Raman, K. uhttps://library.ncl.res.in/content/high-throughput-screening-rna-polymerase-inhibitors-using-fluorescent-utp-analog01444nas a2200193 4500008004100000022001400041245026000055210006900315260011000384300001400494490000600508520048100514100003100995700002701026700002401053700002401077700001901101856013001120 2014 eng d a2046-206900aHighly 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 eval0 aHighly efficient onepot multicomponent synthesis of alphaaminoph aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcJAN a7666-76720 v43 a

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.

1 aMulla, Shafeek, Abdul Rash1 aPathan, Mohsinkhan, Y.1 aChavan, Santosh, S.1 aGample, Suvarna, P.1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/highly-efficient-one-pot-multi-component-synthesis-alpha-aminophosphonates-and-bis-alpha-001370nas a2200145 4500008004100000245011400041210006900155260009000224520069800314100001801012700001901030700002601049700001901075856013001094 2015 eng d00aHigh throughput screening of inhibitors for tuberculosis at NCL : a novel drug discovery initiative from CSIR0 aHigh throughput screening of inhibitors for tuberculosis at NCL  aCSIR-National Chemical Laboratory, PunebCSIR-National Chemical Laboratory, PunecSEP3 a

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

1 aManek, Hardik1 aNawale, Laxman1 aJadhav, Nandadeep, J.1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/high-throughput-screening-inhibitors-tuberculosis-ncl-novel-drug-discovery-initiative-csir02168nas a2200205 4500008004100000245010200041210006900143260000800212490000800220520142500228100002201653700002301675700001701698700002401715700002501739700001901764700002401783700002501807856013001832 2017 eng d00aHybrids of thienopyrimidinones and thiouracils as anti-tubercular agents: SAR and docking studies0 aHybrids of thienopyrimidinones and thiouracils as antitubercular cJAN0 v1273 a

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 7.6–19.1 μg/ml, 12–35 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 11–29 μg/ml, 19–51 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.

1 aPisal, Mahesh, M.1 aNawale, Laxman, U.1 aPatil, Manoj1 aBhansali, Sujit, G.1 aGajbhiye, Jayant, M.1 aSarkar, Dhiman1 aChavan, Subhash, P.1 aBorate, Hanumant, B. uhttps://library.ncl.res.in/content/hybrids-thienopyrimidinones-and-thiouracils-anti-tubercular-agents-sar-and-docking-studies01839nas a2200145 4500008004100000245013400041210006900175260000800244490000800252520125400260100001701514700001901531700001901550856012401569 2019 eng d00aHigh content screening assay for identifying inhibitors against active and dormant state intracellular Mycobacterium tuberculosis0 aHigh content screening assay for identifying inhibitors against  cSEP0 v1643 a

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' 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.

1 aYeware, Amar1 aAgrawal, Sonia1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/high-content-screening-assay-identifying-inhibitors-against-active-and-dormant-state