Aminodehalogenation of 3-chloropyrazine-2-carboxamide with variously substituted benzylamines yielded a series of fifteen 3-benzylaminopyrazine-2-carboxamides. Four compounds possessed in vitro whole cell activity against Mycobacterium tuberculosis H37Rv that was at least equivalent to that of the standard pyrazinamide. MIC values ranged from 6 to 42 μM. The best MIC (6 μM) was displayed by 3-[(4-methylbenzyl)amino]pyrazine-2-carboxamide (8) that also showed low cytotoxicity in the HepG2 cell line (IC50 ≥ 250 μM). Only moderate activity against Enterococcus faecalis and Staphylococcus aureus was observed. No activity was detected against any of tested fungal strains. Molecular docking with mycobacterial enoyl-ACP reductase (InhA) was performed to investigate the possible target of the prepared compounds. Active compounds shared common binding interactions of known InhAinhibitors. Antimycobacterial activity of the title compounds was compared to the previously published benzylamino-substituted pyrazines with differing substitution on the pyrazine core (carbonitrile moiety). The title series possessed comparable activity and lower cytotoxicity than molecules containing a carbonitrile group on the pyrazine ring.

Department of Biological and Medical Sciences Teaching and Research Center of Charles University Faculty of Pharmacy in Hradec Kralove Charles University Zborovska 2089 Hradec Kralove 50003 Czech Republic

Department of Biophysics and Physical Chemistry Faculty of Pharmacy in Hradec Kralove Charles University Akademika Heyrovskeho 1203 8 Hradec Kralove 50005 Czech Republic

Department of Clinical Microbiology University Hospital Hradec Kralove Sokolska 581 Hradec Kralove 50005 Czech Republic

Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Faculty of Pharmacy in Hradec Kralove Charles University Akademika Heyrovskeho 1203 8 Hradec Kralove 50005 Czech Republic

Department of Pharmaceutical Technology Faculty of Pharmacy in Hradec Kralove Charles University Akademika Heyrovskeho 1203 8 Hradec Kralove 50005 Czech Republic

Department of Pharmacology and Toxicology Faculty of Pharmacy in Hradec Kralove Charles University Akademika Heyrovskeho 1203 8 Hradec Kralove 50005 Czech Republic

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World Health Organization . Global Tuberculosis Report 2016. WHO Press; Geneva, Switzerland: 2016. WHO/HTM/TB/2016.13.

Palomino J.C., Martin A. TMC207 becomes bedaquiline, a new anti-TB drug. Future Microbiol. 2013;8:1071–1080. doi: 10.2217/fmb.13.85. PubMed DOI

Zhang Y., Mitchison D. The curious characteristics of pyrazinamide: A review. Int. J. Tuberc. Lung Dis. 2003;7:6–21. PubMed

Zhang Y., Wade M.M., Scorpio A., Zhang H., Sun Z. Mode of action of pyrazinamide: Disruption of Mycobacterium tuberculosis membrane transport and energetics by pyrazinoic acid. J. Antimicrob. Chemother. 2003;52:790–795. doi: 10.1093/jac/dkg446. PubMed DOI

Peterson N.D., Rosen B.C., Dillon N.A., Baughn A.D. Uncoupling environmental pH and intrabacterial acidification from pyrazinamide susceptibility in Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 2015;59:7320–7326. doi: 10.1128/AAC.00967-15. PubMed DOI PMC

Sayahi H., Zimhony O., Jacobs W.R., Shekthman A., Welch J.T. Pyrazinamide, but not pyrazinoic acid, is a competitive inhibitor of NADPH binding to Mycobacterium tuberculosis fatty acid synthase I. Bioorg. Med. Chem. Lett. 2011;21:4804–4807. doi: 10.1016/j.bmcl.2011.06.055. PubMed DOI PMC

Shi W.L., Zhang X.L., Jiang X., Yuan H.M., Lee J.S., Barry C.E., Wang H.H., Zhang W.H., Zhang Y. Pyrazinamide inhibits trans-translation in Mycobacterium tuberculosis. Science. 2011;333:1630–1632. doi: 10.1126/science.1208813. PubMed DOI PMC

Shi W., Chen J., Feng J., Cui P., Zhang S., Weng X., Zhang W., Zhang Y. Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis. Emerg. Microbes Infect. 2014;3:e58. doi: 10.1038/emi.2014.61. PubMed DOI PMC

Zitko J., Dolezal M., Svobodova M., Vejsova M., Kunes J., Kucera R., Jilek P. Synthesis and antimycobacterial properties of N-substituted-6-amino-5-cyanopyrazine-2-carboxamides. Bioorg. Med. Chem. 2011;19:1471–1476. doi: 10.1016/j.bmc.2010.12.054. PubMed DOI

Zitko J., Jampilek J., Dobrovolny L., Svobodova M., Kunes J., Dolezal M. Synthesis and antimycobacterial evaluation of N-substituted 3-aminopyrazine-2,5-dicarbonitriles. Bioorg. Med. Chem. Lett. 2012;22:1598–1601. doi: 10.1016/j.bmcl.2011.12.129. PubMed DOI

Servusova B., Paterova P., Mandikova J., Kubicek V., Kucera R., Kunes J., Dolezal M., Zitko J. Alkylamino derivatives of pyrazinamide: Synthesis and antimycobacterial evaluation. Bioorg. Med. Chem. Lett. 2014;24:450–453. doi: 10.1016/j.bmcl.2013.12.054. PubMed DOI

Zitko J., Servusova B., Janoutova A., Paterova P., Mandikova J., Garaj V., Vejsova M., Marek J., Dolezal M. Synthesis and antimycobacterial evaluation of 5-alkylamino-N-phenylpyrazine-2-carboxamides. Bioorg. Med. Chem. 2015;23:174–183. doi: 10.1016/j.bmc.2014.11.014. PubMed DOI

Servusova-Vanaskova B., Jandourek O., Paterova P., Kordulakova J., Plevakova M., Kubicek V., Kucera R., Garaj V., Naesens L., Kunes J., et al. Alkylamino derivatives of N-benzylpyrazine-2-carboxamide: synthesis and antimycobacterial evaluation. MedChemComm. 2015;6:1311–1317. doi: 10.1039/C5MD00178A. DOI

Servusova B., Eibinova D., Dolezal M., Kubicek V., Paterova P., Pesko M., Kralova K. Substituted N-benzylpyrazine-2-carboxamides: Synthesis and biological evaluation. Molecules. 2012;17:13183–13198. doi: 10.3390/molecules171113183. PubMed DOI PMC

Servusova B., Vobickova J., Paterova P., Kubicek V., Kunes J., Dolezal M., Zitko J. Synthesis and antimycobacterial evaluation of N-substituted 5-chloropyrazine-2-carboxamides. Bioorg. Med. Chem. Lett. 2013;23:3589–3591. doi: 10.1016/j.bmcl.2013.04.021. PubMed DOI

Zitko J., Paterova P., Kubicek V., Mandikova J., Trejtnar F., Kunes J., Dolezal M. Synthesis and antimycobacterial evaluation of pyrazinamide derivatives with benzylamino substitution. Bioorg. Med. Chem. Lett. 2013;23:476–479. doi: 10.1016/j.bmcl.2012.11.052. PubMed DOI

Jandourek O., Dolezal M., Paterova P., Kubicek V., Pesko M., Kunes J., Coffey A., Guo J., Kralova K. N-Substituted 5-amino-6-methylpyrazine-2,3-dicarbonitriles: Microwave-assisted synthesis and biological properties. Molecules. 2014;19:651–671. doi: 10.3390/molecules19010651. PubMed DOI PMC

Hayes B.L. Microwave Synthesis: Chemistry at the Speed of Light. CEM Pub.; Matthews, NC, USA: 2002.

Jandourek O., Dolezal M., Klementova M., Kralova K., Pesko M. Microwave assisted synthesis of new pyrazinamide analogues and their biological evaluation; Proceedings of the 16th International Electronic Conference on Synthetic Organic Chemistry; 29 November 2012; Basel, Switzerland: MDPI; 2012. p. 12.

Kralova K., Pesko M., Paterova P., Kunes J., Tauchman M., Eibinova D., Carillo C., Zitko J., Dolezal M. Substituted N-benzylpyrazine-2-carboxamides, Their Synthesis, Hydro-lipophilic Properties and Evaluation of Their Antimycobacterial and Photosynthesis-inhibiting Activities; Proceedings of the 15th International Electronic Conference on Synthetic Organic Chemistry; 1–30 November 2011; Basel, Switzerland: MDPI; 2011. p. 6.

Kuo M.R., Morbidoni M.R., Alland D., Sneddon S.F., Gourlie B.B., Staveski M.M., Leonard M., Gregory J.S., Janjigian A.D., Yee C., et al. Targeting tuberculosis and malaria through inhibition of enoyl reductase compound activity and structural data. J. Biol. Chem. 2003;278:20851–20859. doi: 10.1074/jbc.M211968200. PubMed DOI

Ghattas M.A., Mansour R.A., Atatreh N., Bryce R.A. Analysis of Enoyl-Acyl Carrier Protein Reductase Structure and Interactions Yields an Efficient Virtual Screening Approach and Suggests a Potential Allosteric Site. Chem. Biol. Drug Des. 2016;87:131–142. doi: 10.1111/cbdd.12635. PubMed DOI

Yang J., Liu Y., Bi J., Cai Q., Liao X., Li W., Guo C., Zhang Q., Lin T., Zhao Y., et al. Structural basis for targeting the ribosomal protein S1 of Mycobacterium tuberculosis by pyrazinamide. Mol. Microbiol. 2015;95:791–803. doi: 10.1111/mmi.12892. PubMed DOI

Petrella S., Gelus-Ziental N., Maudry A., Laurans C., Boudjelloul R., Sougakoff W. Crystal structure of the pyrazinamidase of Mycobacterium tuberculosis: insights into natural and acquired resistance to pyrazinamide. PLoS ONE. 2011;6:e15785. doi: 10.1371/journal.pone.0015785. PubMed DOI PMC

Pandey B., Grover S., Tyagi C., Goyal S., Jamal S., Singh A., Kaur J., Grover A. Molecular principles behind pyrazinamide resistance due to mutations in panD gene in Mycobacterium tuberculosis. Gene. 2016;581:31–42. doi: 10.1016/j.gene.2016.01.024. PubMed DOI

Ciccarelli L., Connell S.R., Enderle M., Mills D.J., Vonck J., Grininger M. Structure and conformational variability of the Mycobacterium tuberculosis fatty acid synthase multienzyme complex. Structure. 2013;21:1251–1257. doi: 10.1016/j.str.2013.04.023. PubMed DOI

Dlabal K., Palat K., Lycka A., Odlerova Z. Synthesis and 1H and 13C NMR spectra of sulfur derivatives of pyrazine derived from amidation product of 2-chloropyrazine and 6-chloro-2-pyrazinecarbonitrile. Tuberculostatic activity. Collect. Czech Chem. Commun. 1990;55:2493–2500. doi: 10.1135/cccc19902493. DOI

Jampilek J., Dolezal M., Kunes J., Satinsky D., Raich I. Novel regioselective preparation of 5-chloropyrazine-2-carbonitrile from pyrazine-2-carboxamide and coupling study of substituted phenylsulfanylpyrazine-2-carboxylic acid derivatives. Curr. Org. Chem. 2005;9:49–60. doi: 10.2174/1385272053369312. DOI

Jandourek O., Dolezal M., Kunes J. Microwave-Assisted Synthesis of Pyrazinamide Derivatives: The Coupling Reaction of 3-Chloropyrazine-2-Carboxamide and Ring-Substituted Anilines. Curr. Org. Synth. 2015;12:189–196. doi: 10.2174/1570179411999141106101501. DOI

Ebeid F.M., Gheit A.K.A., Ezzo E.M., Ali L.I. Decomposition of Triethylamine over Acid Catalysts. J. Chin. Chem. Soc.-Taip. 1982;29:125–129. doi: 10.1002/jccs.198200020. DOI

Dickinson J.M., Mitchison D.A. Observations in vitro on the suitability of pyrazinamide for intermittent chemotherapy of tuberculosis. Tubercle. 1970;51:389–396. doi: 10.1016/0041-3879(70)90004-8. PubMed DOI

McDermott W., Tomsett R. Activation of pyrazinamide and nicotinamide in acidic environments in vitro. Am. Rev. Tuberc. 1954;70:748–754. PubMed

Heifets L., Lindholm-Levy P. Pyrazinamide sterilizing activity in vitro against semidormant Mycobacterium tuberculosis bacterial populations. Am. Rev. Respir. Dis. 1992;145:1223–1225. doi: 10.1164/ajrccm/145.5.1223. PubMed DOI

Butler W.R., Kilburn J.O. Improved method for testing susceptibility of Mycobacterium tuberculosis to pyrazinamide. J. Clin. Microbiol. 1982;16:1106–1109. PubMed PMC

Portaels F., Pattyn S.R. Growth of mycobacteria in relation to the pH of the medium. Ann. Microbiol. 1982;133:213–221. PubMed

Sula J. WHO Co-operative studies on a simple culture technique for the isolation of mycobacteria: 1. Preparation, lyophilization and reconstitution of a simple semi-synthetic concentrated liquid medium; culture technique; growth pattern of different mycobacteria. Bull. Wld. Hlth. Org. 1963;29:589–606. PubMed PMC

Sula J., Sundaresan T.K. WHO Co-operative studies on a simple culture technique for the isolation of mycobacteria: 2. Comparison of the efficacy of lyophilized liquid medium with that of Löwenstein-Jensen (LJ) medium. Bull. Wld. Hlth. Org. 1963;29:607–625. PubMed PMC

Alegre O.S. Estudio comparativo de las posibilidades de los medios de Löwenstein-Jensen y liofilizado reconsititudo de Sula para el aislamiento del Myocbaterium tuberculosis. Bol. Oficina Sanit. Panam. 1967;63:13–16. PubMed

Zitko J., Servusova B., Paterova P., Mandikova J., Kubicek V., Kucera R., Hrabcova V., Kunes J., Soukup O., Dolezal M. Synthesis, antimycobacterial activity and in vitro cytotoxicity of 5-chloro-N-phenylpyrazine-2-carboxamides. Molecules. 2013;18:14807–14825. doi: 10.3390/molecules181214807. PubMed DOI PMC

Bielenica A., Stefańska J., Stępień K., Napiórkowska A., Augustynowicz-Kopeć E., Sanna G., Madeddu S., Boi S., Giliberti G., Wrzosek M., et al. Synthesis, cytotoxicity and antimicrobial activity of thiourea derivatives incorporating 3-(trifluoromethyl) phenyl moiety. Eur. J. Med. Chem. 2015;101:111–125. doi: 10.1016/j.ejmech.2015.06.027. PubMed DOI

Kratky M., Vinsova J., Novotna E., Mandikova J., Trejtnar F., Stolarikova J. Antibacterial activity of salicylanilide 4-(trifluoromethyl)-benzoates. Molecules. 2013;18:3674–3688. doi: 10.3390/molecules18043674. PubMed DOI PMC

Keir W.F., MacLennan A.H., Wood H.C. Amidinoacetamides in the synthesis of pyrazines and pteridines. J. Chem. Soc. Perk. T. 1. 1977;11:1321–1325. doi: 10.1039/p19770001321. DOI

Franzblau S.G., Witzig R.S., McLaughlin J.C., Torres P., Madico G., Hernandez A., Degnan M.T., Cook M.B., Quenzer V.K., Ferguson R.M., et al. Rapid, low-technology MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar Blue assay. J. Clin. Microbiol. 1998;36:362–366. PubMed PMC

Jones R.N., Barry A.L. Optimal dilution susceptibility testing conditions, recommendations for MIC interpretation, and quality control guidelines for the ampicillin-sulbactam combination. J. Clin. Microbiol. 1987;25:1920–1925. PubMed PMC

National Committee for Clinical Laboratory Standards . Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts: Proposed Standard M 27-P. National Committee for Clinical Laboratory Standards; Villanova, PA, USA: 1992.

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