Mutagenicity Assessment Of A Benzopentathiepine Derivative In Ames Test

V. Bykov, head of Pharmacological Research Department, I. Sedykh, Ph.D. in Biology, Deputy President for general issues, A. Bykova, junior scientist LLC Innovative Pharmacology Research 634021, Russia, Tomsk, Elizarovikh str., 79/4 Е-mail: preclin5_dep@iphar.ru

Abstract

Summary. Assessment of mutagenic properties of a new drug is an important part of its preclinical trials, because allows to study the drug’s ability to induce different mutations in fetal and somatic cells. An important aspect of mutagenicity assessment of a new drug is its structural similarity to the known mutagens. This structural similarity is analyzed with the aid of a large database of mutagenic properties of a wide variety of chemical compounds and with special computer programs. Experimental assessment of mutagenicity of new drug candidates requires the use of a battery of at least 2 tests, including an in vivo chromosomal damage test and a genetic mutation induction test, for which Ames test is usually used, which an accepted alternative to animal testing. On one hand, replacement of animals with less developed organisms is one of the 3R principles, on the other hand, using bacterial strains allows to screen compounds and quickly assess the potential genotoxicity of a compound. The obtained data can then be used to properly design subsequent toxicity tests. The present study assessed the ability of the test compound to induce mutations in S. typhimurium (ТА98, ТА100, ТА1535, ТА1537) and E. сoli (WP2 uvrA and WP2/pKM101) strains in six concentrations: 15,6; 31,2; 62,4; 125,0; 250,0 and 500,0 μg/ml. To assess potential genotoxicity of the test compound and its metabolites, we have performed testing with metabolic activation in the presence of rat liver S9 fraction (Aroclor-1254 inducer) and without it. The study has demonstrated the absence of mutagenic properties of the test compounds in all used bacterial strains, however, we can’t rule out possible genotoxicity after multiple administration.

References

  1. Freeman B.A., Crapo J.D. Free radical and tissue injury. Adv. Biol. Disease. 1984; 1: 26–40.
  2. Barnes D.E., Lindahl T., Segwick B. DNA Repair. Curr. Opin. Cell Biol. 1993; 5: 424–33.
  3. Guengerich, F.P. Metabolic activation of carcinogens. Pharmac. Ther., 1992; 54: 17–61.
  4. Miller E.C. and Miller J.A. Searches for ultimate carcinogens and their reaction with cellular macromolecules. Cancer. 1981; 47: 2327–45.
  5. Rukovodstvo po provedeniyu doklinicheskih issledovaniy lekarstvennyh sredstv, ch. I.
  6. Zhengyin Yan, Gary Caldwell. Optimization in drug discovery: in vitro methods. Chapter «Improvement of the Ames test using human liver S9 preparation». Methods in pharmacology and toxicology. Humana Press, 2004.
  7. Venitt S., Bartsch H., Becking G. et. al. Short-term assays using bacteria. In: LongTerm and Short-Term Assays for Carcinogens: A Critical Appraisal. IARC Scientific Publications, no. 83, Lyon, Chap. 5, 1986. Oxford University Press, Oxford, UK: 143–56.
  8. Anderson, D. Genotoxicity assays. In: Molecular Aspects of Oxidative Drug Metabolizing Enzymes. Arinç, E., Schenkman, J.B., and Hodgson, E., Eds., NATO ASI Series, 1995; H90, Springer-Verlag, Berlin: 303–96.
  9. Austin E.A., Graves J.M., Hite L.A., Parker C.T., and Schnaitman C.A. Genetic analyses of lipopolysaccharide core biosynthesis by Escherichia coli K12: insertion mutagenesis of th rfa locus. J. Bacteriol. 1990; 172: 5312–25.
  10. Parker C.T., Kloser A.W., Schnaitman C.A., Stein M.A., Gottesman S., and Gibson B.W. Role of the rfaG and rfaP genes in determining the lipopolysaccharide core structure and cell surface propertie s of Escherichia coli K-12. 1992.
  11. Anderson D. Genotoxicity assays. In: Molecular Aspects of Oxidative Drug Metabolizing Enzymes.
  12. Arinc E., Schenkman J.B. and Hodgson E., Eds., NATO ASI Series. 1995; H90, Springer-Verlag, Berlin: 303–96.
  13. Maron D.M. and Ames B.N. Revised methods for Salmonella mutagenicity test. Mutat. Res. 1983; 113: 173–215.
  14. Khomenko T.N., Korchagina D.V., Komarova N.I., Volcho K.P., Salakhutdinov N.F. Synthesis of 6-Amino-benzopentathiepines by Reactions of 4-Nitro-benzyldithiol-2-ones with NaHS Lett. Org. Chem. 2011; 8 (3): 193–7.
  15. Kulikova E.A., Volcho K.P., Salakhutdinov N.F., Kulikov A.V. Benzopentathiepine Derivative, 8-(Trifluoromethyl)-1,2,3,4,5-Benzopentathiepin-6-Amine Hydrochloride (TC-2153), as a Promising Anti-depressant of New Generation Letters in Drug Design & Discovery. 2017; 14; 8: 974–84 DOI:10.2174/1570180814666161121112417.
  16. Davidson B.S.; Molinski T.F.; Barrows L.R.; Ireland C.M. J. Am. Chem. Soc. 1991; 113: 4709–10.
  17. Barrows L.R.; Paxton M.B.; Kennedy K.A.; Thompson L.H. Carcinogenesis. 1991; 12: 805.
  18. Sato R.; Ohyama T.; Ogawa S. Heterocycles 1995; 416 893–6.
  19. Ang E.L., Obbard J.P., Zhao H.M. Probing the molecular determinants of aniline dioxygenase substrate specificity by saturation mutagenesis. FEBS J. 2007; 274: 928–93.
  20. Gus`kova T.D. Ocenka bezopasnosti lekarstvennyh sredstv na stadii doklinicheskogo izucheniya. Himiko-farmacevtich. Zhurn., 1990; 7: 10–5.
  21. OECD Bacterial Reverse Mutation Test 471, Adopted 21st July. 1997.
  22. Instrukciya po primeneniyu testa E`ymsa v mikroplanshetnom formate Ames MPF™ Penta I. S. typhimurium TA98, TA100, TA1535, TA1537, E. coli uvrA + E. coli /pKM101. Xenometrix. Versiya 5.5, yanvar` 2017.
  23. Edwards D.I. Reduction of nitroimidazoles and DNA damage. Biochem Pharmacol. 1986; 35: 53–8.
  24. Lopez Nigro M.M., Carballo M.A. Genotoxicity and cell death induced by tinidazole (TNZ). ToxicolLett. 2008; 180: 46–52.
  25. Alja Plosnik, Marjan Vracko, Marija Sollner Dolenc, Mutagenic and carcinogenic structural alerts and their mechanisms of action, Plosnik A, Vracko M, Sollner Dolenc M. Mutagenic and carcinogenic structural alerts and their mechanisms of action Arh Hig Rada Toksikol 2016; 67: 169–82.
  26. Koblyakov V.A. Induktory supersemeystva citohroma p450 kak promotory kancerogeneza. Biohimiya, 1998; 63 (8): 1043–58.

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