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Screening and Development of New Inhibitors of Ftsz from M University of Dundee Screening and development of new inhibitors of FtsZ from M. Tuberculosis Mathew, Bini; Hobrath, Judith Varady; Ross, Larry; Connelly, Michele C.; Lofton, Hava; Rajagopalan, Malini Published in: PLoS ONE DOI: 10.1371/journal.pone.0164100 Publication date: 2016 Licence: CC BY Document Version Publisher's PDF, also known as Version of record Link to publication in Discovery Research Portal Citation for published version (APA): Mathew, B., Hobrath, J. V., Ross, L., Connelly, M. C., Lofton, H., Rajagopalan, M., Guy, R. K., & Reynolds, R. C. (2016). Screening and development of new inhibitors of FtsZ from M. Tuberculosis. PLoS ONE, 11(10), [e0164100]. https://doi.org/10.1371/journal.pone.0164100 General rights Copyright and moral rights for the publications made accessible in Discovery Research Portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from Discovery Research Portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain. • You may freely distribute the URL identifying the publication in the public portal. Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Download date: 09. Oct. 2021 RESEARCH ARTICLE Screening and Development of New Inhibitors of FtsZ from M. Tuberculosis Bini Mathew3, Judith Varady Hobrath4, Larry Ross3, Michele C. Connelly5, Hava Lofton6¤, Malini Rajagopalan6, R. Kiplin Guy5, Robert C. Reynolds1,2* 1 Department of Chemistry, The University of Alabama at Birmingham, Birmingham, Alabama, 35294, United States of America, 2 Division of Hematology and Oncology, The University of Alabama at Birmingham, Birmingham, Alabama, 35294, United States of America, 3 Drug Discovery Division, Southern Research Institute, 2000 Ninth Avenue South, Birmingham, Alabama, 35205, United States of America, 4 Drug Discovery Unit, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, United Kingdom, a11111 5 Dept. Chemical Biology & Therapeutics, St Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN, 38105, United States of America, 6 The University of Texas Health Science Center at Tyler, Tyler, Texas, 75708, United States of America ¤ Current address: Department of Medical Biochemistry and Microbiology, Uppsala University, SE-75123, Uppsala, Sweden * [email protected] OPEN ACCESS Citation: Mathew B, Hobrath JV, Ross L, Connelly Abstract MC, Lofton H, Rajagopalan M, et al. (2016) Screening and Development of New Inhibitors of A variety of commercial analogs and a newer series of Sulindac derivatives were screened FtsZ from M. Tuberculosis. PLoS ONE 11(10): for inhibition of M. tuberculosis (Mtb) in vitro and specifically as inhibitors of the essential e0164100. doi:10.1371/journal.pone.0164100 mycobacterial tubulin homolog, FtsZ. Due to the ease of preparing diverse analogs and a Editor: Selvakumar Subbian, Rutgers Biomedical favorable in vivo pharmacokinetic and toxicity profile of a representative analog, the Sulin- and Health Sciences, UNITED STATES dac scaffold may be useful for further development against Mtb with respect to in vitro bac- Received: May 13, 2016 terial growth inhibition and selective activity for Mtb FtsZ versus mammalian tubulin. Accepted: September 20, 2016 Further discovery efforts will require separating reported mammalian cell activity from both antibacterial activity and inhibition of Mtb FtsZ. Modeling studies suggest that these ana- Published: October 21, 2016 logs bind in a specific region of the Mtb FtsZ polymer that differs from human tubulin and, in Copyright: © 2016 Mathew et al. This is an open combination with a pharmacophore model presented herein, future hybrid analogs of the access article distributed under the terms of the Creative Commons Attribution License, which reported active molecules that more efficiently bind in this pocket may improve antibacterial permits unrestricted use, distribution, and activity while improving other drug characteristics. reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information Introduction files. Funding: Primary support for this study was Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is one of the most prevalent provided through the National Institutes of Health infectious diseases worldwide. A recent report by the World Health Organization (WHO) (http://www.nih.gov/) grants 1 R01 AI50470-01A1 reveals that in 2014 there were an estimated 9.6 million new cases of TB (12% co-infected with (P.I.: RCR) and R01 AI48417 (P.I.: MR). This work HIV) and 1.5 million people died from the disease, including 1.1 million deaths among HIV- was supported in part by the National Cancer negative individuals and 400,000 among people who were HIV-positive.[1] Moreover, the Institute (http://www.cancer.gov) grant 1 R01 increasing development of multi-drug resistant TB (MDR-TB) strains, forms of TB that do not CA131378 (P.I.: RCR) and also by the United States Department of Defense (http://www. respond to standard front line regimens, is a serious burden on public health systems through- defense.gov) Era of Hope Idea Award W81XWH- out the world. More recently, extensively drug resistant tuberculosis (XDR-TB) that is also 07-1-0463 (P.I.: RCR) through supplying samples. resistant to second line drugs has emerged in numerous areas. These stark realities emphasize PLOS ONE | DOI:10.1371/journal.pone.0164100 October 21, 2016 1 / 23 Screening and Development of New FtsZ Inhibitors for M. Tuberculosis The funders had no role in study design, data the acute need for constant reinvention of the anti-TB drug pipeline targeting novel, alternative collection and analysis, decision to publish, or drug pathways.[2–3] preparation of the manuscript. New targets for tuberculosis are regularly being identified from existing genomic data and Competing Interests: The authors have declared new screening techniques.[3] The bacterial tubulin homolog, FtsZ (Filamenting temperature- that no competing interests exist. sensitive protein Z), is essential for bacterial cell division and is an ideal target for novel antimi- crobials. In recent years, FtsZ has received considerable attention for the discovery of novel antibacterial and anti-TB drugs.[4–17] In our search for new active scaffolds against Mtb FtsZ, a variety of small molecules reported to have antibacterial activity, and, furthermore, some of which were considered to inhibit Mtb or other bacterial FtsZs were acquired and screened for both antitubercular activity and Mtb FtsZ inhibition. A consistent set of antibacterial activity data in parallel with FtsZ screening results should be useful to prioritize active scaffolds for new analog optimization. Furthermore, the potent combination of a new crystal structure and these activity data will allow advance- ment of robust consensus binding models that should help medicinal chemists enhance selec- tive activity against the bacterial protein target and whole bacteria while potentially minimizing off-target effects against the direct mammalian homolog, tubulin, as well as reduc- ing mammalian toxicity through other off-target activities. Beyond the aforementioned anti- bacterial/FtsZ actives or related compounds, we were particularly intrigued by the reported similarities of certain non-steroidal anti-inflammatory drugs (NSAIDs), e.g. Indomethacin and Sulindac analogs, to the known tubulin polymerization inhibitor Colchicine.[18,19] Colchicine has been reported to be one of the few known tubulin inhibitors that demonstrates activity against Mtb FtsZ.[15] Sulindac belongs to this chemically diverse group and, importantly, is not overtly toxic in vivo but shows clinical efficacy for longer term treatment regimens in can- cer chemoprevention.[20–23] The NSAIDs are excellent pharmacophores showing good activ- ity through animal models and in the clinic for numerous indications. As part of an ongoing program to study the chemical biology of interesting NSAID scaffolds such as Sulindac, we have investigated a variety of analogs and their on-target (COX-1 and 2) and off-target (e.g. cell cytotoxicity, PDE5, PDE10A) activities.[24–25] Among the interesting and atypical activi- ties of the NSAIDs, certain known drugs (e.g. Ibuprofen, Aspirin) have been reported to show antibacterial activity.[26–30] An indomethacine analog closely related to sulindac sulfide amide (SSA) has been reported to inhibit tubulin polymerization in vitro in a dose response manner.[18] Hence, we added this early lead Sulindac analog to our initial anti-TB/FtsZ assays and confirmed that it is a modest potency inhibitor of Mtb FtsZ while showing no inhibition of human tubulin at 100 μM concentration. The activity of this initial lead warrants the explora- tion of new sulindac analog series against FtsZ. Herein, we report the screening of a number of acquired and synthesized samples and a lead Sulindac analog available in our labs against FtsZ from M. tuberculosis. These compounds advanced in a series of assays depending on activity that include Mtb FtsZ, Mtb H37Ra, MAC NJ211 and/or Mtb H37Rv, tubulin polymerization,
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