Cyto‑genotoxicity Assessment of Potential Anti‑tubercular Drug Candidate Molecule‑trans‑cyclohexane‑1, 4‑diamine Derivative‑9u in Human Lung Epithelial Cells A549


Affiliations

  • Atma Ram Sanatan Dharma College, New Delhi, India
  • Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India
  • Kumaun University, Department of Pharmaceutical Sciences, Nainital
  • Atma Ram Sanatan Dharma College, New Delhi
  • Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Dehradun, Uttarakhand, India

Abstract

Increasing incidences of multiple drug‑resistance (MDR) in Mycobacterium tuberculosis are emerging as one among the serious public health threats and socio‑economic burden to the third world countries including India. Last couples of decades are witnesses of the dedicated and sustained efforts made toward the development of target specific and cost‑effective antimicrobial agents against MDR‑M. tuberculosis. However, the drugs in use are still incapable of controlling the upsurge of MDR. Thus, in order to address the issue, we synthesized a library of symmetrical trans‑cyclohexane‑1, 4‑diamine derivatives and evaluated their anti‑mycobacterium activity in H37RV strain of M. tuberculosis. A range of efficacy has been recorded in different derivatives of synthesized compounds and compound “9u” having i‑propyl group substitution at p‑position, was found to have more significant detrimental effects against the tested strain of M. tuberculosis. The present investigations were aimed to study whether the effective anti‑mycobacterium concentrations of “9u” are biologically safe to human cells or not? The human lung epithelial cell line‑A549 were exposed to a range of concentrations, i.e., at and above the anti‑mycobacterium effective dose of “9u” for a period of 0‑96 h. The standard endpoints of cytotoxicity viz., tetrazolium bromide salt (3‑[4,5‑dimethylthiazol‑2‑yl]‑2,5‑diphenyl tetrazolium bromide), neutral red uptake, lactate dehydrogenase release, trypan blue dye exclusion assays; and genotoxicity viz., micronucleus and chromosomal aberrations assays were used to evaluate the bio‑safety of test compound. The compound “9u” shows no significant cytotoxicity and genotoxicity in A549 cells exposed to 10 − 5 M for 72 h, a concentration substantially higher than the concentration kill the H37Rv strain of M. tuberculosis. The compound 9u was found to be safe up to 10 − 4 M if given for 24 h. The data reveal the therapeutic potential of compound 9u against M. tuberculosis without any having any cytotoxicity and genotoxicity responses.

Keywords

A549 cell line, chromosomal aberration, cytotoxicity, micronucleus assay, Mycobacterium tuberculosis

Full Text:

References

WHO. Global Tuberculosis Report. WHO/HTM/TB/2013.15 : IX‑XI; 2013.

Patil SD, Angadi KM, Modak MS, Bodhankar MG. Studies on drug‑resistance pattern by phenotypic methods in Mycobacterium tuberculosis isolates in a tertiary care hospital. Int J Microbiol Res 2013;5:497‑501.

Michalska K, Karpiuk I, Król M, Tyski S. Recent development of potent analogues of oxazolidinone antibacterial agents. Bioorg Med Chem 2013;21:577‑91.

Russell DG. Mycobacterium tuberculosis: Here today, and here tomorrow. Nat Rev Mol Cell Biol 2001;2:569‑77.

Clemens DL. Characterization of the Mycobacterium tuberculosis phagosome. Trends Microbiol 1996;4:113‑8.

Deretic V, Fratti RA. Mycobacterium tuberculosis phagosome. Mol Microbiol 1999;31:1603‑9.

Vergne I, Chua J, Deretic V. Mycobacterium tuberculosis phagosome maturation arrest: Selective targeting of PI3P‑dependent membrane trafficking. Traffic 2003;4:600‑6.

Sinha K, Marak IT, Singh WA. Adverse drug reactions in tuberculosis patients due to directly observed treatment strategy therapy: Experience at an outpatient clinic of a teaching hospital in the city of Imphal, Manipur, India. J Assoc Chest Physicians. 2013:1:50‑53.

Manca ML, Cassano R, Valenti D, Trombino S, Ferrarelli T, Picci N, et al. Isoniazid‑gelatin conjugate microparticles containing rifampicin for the treatment of tuberculosis. J Pharm Pharmacol 2013;65:1302‑11.

Clemens DL, Lee BY, Xue M, Thomas CR, Meng H, Ferris D, et al. Targeted intracellular delivery of antituberculosis drugs to Mycobacterium tuberculosis‑infected macrophages via functionalized mesoporous silica nanoparticles. Antimicrob Agents Chemother 2012;56:2535‑45.

Dye C, Espinal MA, Watt CJ, Mbiaga C, Williams BG. Worldwide incidence of multidrug‑resistant tuberculosis. J Infect Dis 2002;185:1197‑202.

Farmer P, Kim JY. Community based approaches to the control of multidrug resistant tuberculosis: Introducing “DOTS‑plus”. BMJ 1998;317:671‑4.

Cantwell MF, Snider DE Jr, Cauthen GM, Onorato IM. Epidemiology of tuberculosis in the United States, 1985 through 1992. JAMA 1994;272:535‑9.

McIlleron H, Meintjes G, Burman WJ, Maartens G. Complications of antiretroviral therapy in patients with tuberculosis: Drug interactions, toxicity, and immune reconstitution inflammatory syndrome. J Infect Dis 2007;196 Suppl 1:S63‑75.

Beena, Joshi S, Kumar N, Kidwai S, Singh R, Rawat DS. Synthesis and antitubercular activity evaluation of novel unsymmetrical cyclohexane‑1,2‑diamine derivatives. Arch Pharm (Weinheim) 2012;345:896‑901.

Sharma M, Joshi P, Kumar N, Joshi S, Rohilla RK, Roy N, et al. Synthesis, antimicrobial activity and structure‑activity relationship study of N, N‑dibenzyl‑cyclohexane‑1,2‑diamine derivatives. Eur J Med Chem 2011;46:480‑7.

Sharma M, Roy N, Rohilla RK, Rawat DS. Indian Patent Appl No: 1462/DEL; 2008.

Kumar D, Joshi S, Rohilla RK, Roy N, Rawat DS. Synthesis and antibacterial activity of benzyl‑[3‑(benzylamino‑methyl)‑c yclohexylmethyl]‑amine derivatives. Bioorg Med Chem Lett 2010;20:893‑5.

Kumar N, Kapoor E, Singh R, Kidwai S, Kumbukgolla W, Bhagat S, et al. Synthesis and antibacterial/antitubercular activity evaluation of symmetrical trans‑cyclohexane‑1,4‑diamine derivatives. Indian J Chem 2013;52B: 1441‑50.

Kashyap MP, Singh AK, Siddiqui MA, Kumar V, Tripathi VK, Khanna VK, et al. Caspase cascade regulated mitochondria mediated apoptosis in monocrotophos exposed PC12 cells. Chem Res Toxicol 2010;23:1663‑72.

Siddiqui MA, Singh G, Kashyap MP, Khanna VK, Yadav S, Chandra D, et al. Influence of cytotoxic doses of 4‑hydroxynonenal on selected neurotransmitter receptors in PC‑12 cells. Toxicol in vitro 2008;22:1681‑8.

Pant AB, Agarwal AK, Sharma VP, Seth PK. In vitro cytotoxicity evaluation of plastic biomedical devices. Hum Exp Toxicol 2001;20:412‑7.

Srivastava RK, Lohani M, Pant AB, Rahman Q. Cyto‑genotoxicity of amphibole asbestos fibers in cultured human lung epithelial cell line: Role of surface iron. Toxicol Ind Health 2010;26:575‑82.

Li AP, Lu C, Brent JA, Pham C, Fackett A, Ruegg CE, et al. Cryopreserved human hepatocytes: Characterization of drug‑metabolizing enzyme activities and applications in higher throughput screeningassays for hepatotoxicity, metabolic stability, and drug‑drug interaction potential. Chem Biol Interact 1999;121:17‑35.

Heussner AH, Dietrich DR, O’Brien E. In vitro investigation of individual and combined cytotoxic effects of ochratoxin A and other selected mycotoxins on renal cells. Toxicol in vitro 2006;20:332‑41.

White RE. High‑throughput screening in drug metabolism and pharmacokinetic support of drug discovery. Annu Rev Pharmacol Toxicol 2000;40:133‑57.

Ní Shúilleabháin S, Mothersill C, Sheehan D, O’Brien NM, O’ Halloran J, Van Pelt FN, et al. In vitro cytotoxicity testing of three zinc metal salts using established fish cell lines. Toxicol in vitro 2004;18:365‑76.

Ehret R, Baumann W, Brischwein M, Schwinde A, Stegbauer K, Wolf B. Monitoring of cellular behaviour by impedance measurements on interdigitated electrode structures. Biosens Bioelectron 1997;12:29‑41.

Vahdati‑Mashhadian N, Jaafari MR, Nosrati A. Differential toxicity of rifampin on HepG2 and Hep2 cells using MTT test and electron microscope. Pharmacologyonline 2007;3:405‑13.

Isefuku S, Joyner CJ, Simpson AH. Toxic effect of rifampicin on human osteoblast‑like cells. J Orthop Res 2001;19:950‑4.

United States Pharmacopeial Convention Inc. USPDI, Drug Information for the Health Care Professional. Vol. 1. Englewood, USA: Micromedex; 2001. p. 2597.

Knasmüller S, Parzefall W, Sanyal R, Ecker S, Schwab C, Uhl M, et al. Use of metabolically competent human hepatoma cells for the detection of mutagens and antimutagens. Mutat Res 1998;402:185‑202.

Strolin Benedetti M, Dostert P. Induction and autoinduction properties of rifamycin derivatives: A review of animal and human studies. Environ Health Perspect 1994;102 Suppl 9:101‑5.

Olufunsho A, Alade A. Rifampicin: Cytotoxic and genotoxic action. Niger J Health Biomed Sci 2010;9:2.

Masjedi MR, Heidary A, Mohammadi F, Velayati AA, Dokouhaki P. Chromosomal aberrations and micronuclei in lymphocytes of patients before and after exposure to anti‑tuberculosis drugs. Mutagenesis 2000;15:489‑94.

Russell DG, Mwandumba HC, Rhoades EE. Mycobacterium and the coat of many lipids. J Cell Biol 2002;158:421‑6.

Ernst JD. Macrophage receptors for Mycobacterium tuberculosis. Infect Immun 1998;66:1277‑81.

Cegielski JP. Extensively drug‑resistant tuberculosis: “there must be some kind of way out of here”. Clin Infect Dis 2010;50 Suppl 3:S195‑200.

Grosset JH, Singer TG, Bishai WR. New drugs for the treatment of tuberculosis: Hope and reality. Int J Tuberc Lung Dis 2012;16:1005‑14.

Mitchison D, Davies G. The chemotherapy of tuberculosis: Past, present and future. Int J Tuberc Lung Dis 2012;16:724‑32.

Kolpakova TA, Kolpakov MA, Bashkirova IuV, Rachkovskaia LN, Burylin SIu, Liubarskiĭ MS. Effects of the enterosorbent SUMS‑1 on isoniazid pharmacokinetics and lipid peroxidation in patients with pulmonary tuberculosis and drug‑induced hepatic lesions. Probl Tuberk 2001; 3:34‑6.

de Souza AF, de Oliveira e Silva A, Baldi J, de Souza TN, Rizzo PM. Hepatic functional changes induced by the combined use of isoniazid, pyrazinamide and rifampicin in the treatment of pulmonary tuberculosis. Arq Gastroenterol 1996;33:194‑200.


Refbacks

  • There are currently no refbacks.