Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1993 Oct;37(10):2179–2186. doi: 10.1128/aac.37.10.2179

Drug features that contribute to the activity of quinolones against mammalian topoisomerase II and cultured cells: correlation between enhancement of enzyme-mediated DNA cleavage in vitro and cytotoxic potential.

S H Elsea 1, P R McGuirk 1, T D Gootz 1, M Moynihan 1, N Osheroff 1
PMCID: PMC192247  PMID: 8257142

Abstract

CP-115,953 [6,8-difluoro-7-(4'-hydroxyphenyl)-1-cyclopropyl-4- quinolone-3-carboxylic acid] is a novel quinolone that is highly active against topoisomerase II in vitro and in mammalian cells in culture (M. J. Robinson, B. A. Martin, T. D. Gootz, P. R. McGuirk, M. Moynihan, J. A. Sutcliffe, and N. Osheroff, J. Biol. Chem. 266:14585-14592, 1991). However, the features of the drug that contribute to its activity towards mammalian systems have not been characterized. Therefore, CP-115,953 and a series of related quinolones were examined for their activity against calf thymus topoisomerase II and cultured mammalian cells. CP-115,953 stimulated DNA cleavage mediated by the type II enzyme with a potency that was approximately 600-fold greater than that of the antimicrobial quinolone ciprofloxacin and approximately 50-fold greater than that of the antineoplastic drug etoposide. As determined by the ability to enhance enzyme-mediated DNA cleavage, quinolone activity towards calf thymus topoisomerase II was enhanced by the presence of a cyclopropyl group at the N-1 ring position and by the presence of a fluorine at C-8. Furthermore, the 4'-hydroxyphenyl substituent at the C-7 position was critical for the potency of CP-115,953 towards the mammalian type II enzyme. In this regard, the aromatic nature of the C-7 ring as well as the presence and the position of the 4'-hydroxyl group contributed greatly to drug activity. Finally, the cytotoxicity of quinolones in the CP-115,953 series towards mammalian cells paralleled the in vitro stimulation of DNA cleavage by topoisomerase II rather than the inhibition of enzyme-catalyzed DNA relaxation. This correlation strongly suggests that these quinolones promote cell death by converting topoisomerase II to a cellular poison.

Full text

PDF
2179

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andersen A. H., Christiansen K., Zechiedrich E. L., Jensen P. S., Osheroff N., Westergaard O. Strand specificity of the topoisomerase II mediated double-stranded DNA cleavage reaction. Biochemistry. 1989 Jul 25;28(15):6237–6244. doi: 10.1021/bi00441a015. [DOI] [PubMed] [Google Scholar]
  2. Anderson A. H., Sørensen B. S., Christiansen K., Svejstrup J. Q., Lund K., Westergaard O. Studies of the topoisomerase II-mediated cleavage and religation reactions by use of a suicidal double-stranded DNA substrate. J Biol Chem. 1991 May 15;266(14):9203–9210. [PubMed] [Google Scholar]
  3. Barrett J. F., Gootz T. D., McGuirk P. R., Farrell C. A., Sokolowski S. A. Use of in vitro topoisomerase II assays for studying quinolone antibacterial agents. Antimicrob Agents Chemother. 1989 Oct;33(10):1697–1703. doi: 10.1128/aac.33.10.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Domagala J. M., Bridges A. J., Culbertson T. P., Gambino L., Hagen S. E., Karrick G., Porter K., Sanchez J. P., Sesnie J. A., Spense F. G. Synthesis and biological activity of 5-amino- and 5-hydroxyquinolones, and the overwhelming influence of the remote N1-substituent in determining the structure-activity relationship. J Med Chem. 1991 Mar;34(3):1142–1154. doi: 10.1021/jm00107a039. [DOI] [PubMed] [Google Scholar]
  5. Domagala J. M., Hanna L. D., Heifetz C. L., Hutt M. P., Mich T. F., Sanchez J. P., Solomon M. New structure-activity relationships of the quinolone antibacterials using the target enzyme. The development and application of a DNA gyrase assay. J Med Chem. 1986 Mar;29(3):394–404. doi: 10.1021/jm00153a015. [DOI] [PubMed] [Google Scholar]
  6. Elsea S. H., Osheroff N., Nitiss J. L. Cytotoxicity of quinolones toward eukaryotic cells. Identification of topoisomerase II as the primary cellular target for the quinolone CP-115,953 in yeast. J Biol Chem. 1992 Jul 5;267(19):13150–13153. [PubMed] [Google Scholar]
  7. Froelich-Ammon S. J., McGuirk P. R., Gootz T. D., Jefson M. R., Osheroff N. Novel 1-8-bridged chiral quinolones with activity against topoisomerase II: stereospecificity of the eukaryotic enzyme. Antimicrob Agents Chemother. 1993 Apr;37(4):646–651. doi: 10.1128/aac.37.4.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gale K. C., Osheroff N. Uncoupling the DNA cleavage and religation activities of topoisomerase II with a single-stranded nucleic acid substrate: evidence for an active enzyme-cleaved DNA intermediate. Biochemistry. 1990 Oct 16;29(41):9538–9545. doi: 10.1021/bi00493a007. [DOI] [PubMed] [Google Scholar]
  9. Glisson B., Gupta R., Smallwood-Kentro S., Ross W. Characterization of acquired epipodophyllotoxin resistance in a Chinese hamster ovary cell line: loss of drug-stimulated DNA cleavage activity. Cancer Res. 1986 Apr;46(4 Pt 2):1934–1938. [PubMed] [Google Scholar]
  10. Hooper D. C., Wolfson J. S. Fluoroquinolone antimicrobial agents. N Engl J Med. 1991 Feb 7;324(6):384–394. doi: 10.1056/NEJM199102073240606. [DOI] [PubMed] [Google Scholar]
  11. Hoshino K., Sato K., Akahane K., Yoshida A., Hayakawa I., Sato M., Une T., Osada Y. Significance of the methyl group on the oxazine ring of ofloxacin derivatives in the inhibition of bacterial and mammalian type II topoisomerases. Antimicrob Agents Chemother. 1991 Feb;35(2):309–312. doi: 10.1128/aac.35.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoshino K., Sato K., Une T., Osada Y. Inhibitory effects of quinolones on DNA gyrase of Escherichia coli and topoisomerase II of fetal calf thymus. Antimicrob Agents Chemother. 1989 Oct;33(10):1816–1818. doi: 10.1128/aac.33.10.1816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hussy P., Maass G., Tümmler B., Grosse F., Schomburg U. Effect of 4-quinolones and novobiocin on calf thymus DNA polymerase alpha primase complex, topoisomerases I and II, and growth of mammalian lymphoblasts. Antimicrob Agents Chemother. 1986 Jun;29(6):1073–1078. doi: 10.1128/aac.29.6.1073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kohlbrenner W. E., Wideburg N., Weigl D., Saldivar A., Chu D. T. Induction of calf thymus topoisomerase II-mediated DNA breakage by the antibacterial isothiazoloquinolones A-65281 and A-65282. Antimicrob Agents Chemother. 1992 Jan;36(1):81–86. doi: 10.1128/aac.36.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kreuzer K. N., Cozzarelli N. R. Escherichia coli mutants thermosensitive for deoxyribonucleic acid gyrase subunit A: effects on deoxyribonucleic acid replication, transcription, and bacteriophage growth. J Bacteriol. 1979 Nov;140(2):424–435. doi: 10.1128/jb.140.2.424-435.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Liu L. F. DNA topoisomerase poisons as antitumor drugs. Annu Rev Biochem. 1989;58:351–375. doi: 10.1146/annurev.bi.58.070189.002031. [DOI] [PubMed] [Google Scholar]
  17. Maxwell A. The molecular basis of quinolone action. J Antimicrob Chemother. 1992 Oct;30(4):409–414. doi: 10.1093/jac/30.4.409. [DOI] [PubMed] [Google Scholar]
  18. Miller K. G., Liu L. F., Englund P. T. A homogeneous type II DNA topoisomerase from HeLa cell nuclei. J Biol Chem. 1981 Sep 10;256(17):9334–9339. [PubMed] [Google Scholar]
  19. Moreau N. J., Robaux H., Baron L., Tabary X. Inhibitory effects of quinolones on pro- and eucaryotic DNA topoisomerases I and II. Antimicrob Agents Chemother. 1990 Oct;34(10):1955–1960. doi: 10.1128/aac.34.10.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oomori Y., Yasue T., Aoyama H., Hirai K., Suzue S., Yokota T. Effects of fleroxacin on HeLa cell functions and topoisomerase II. J Antimicrob Chemother. 1988 Oct;22 (Suppl 500):91–97. doi: 10.1093/jac/22.supplement_d.91. [DOI] [PubMed] [Google Scholar]
  21. Osheroff N. Effect of antineoplastic agents on the DNA cleavage/religation reaction of eukaryotic topoisomerase II: inhibition of DNA religation by etoposide. Biochemistry. 1989 Jul 25;28(15):6157–6160. doi: 10.1021/bi00441a005. [DOI] [PubMed] [Google Scholar]
  22. Osheroff N., Shelton E. R., Brutlag D. L. DNA topoisomerase II from Drosophila melanogaster. Relaxation of supercoiled DNA. J Biol Chem. 1983 Aug 10;258(15):9536–9543. [PubMed] [Google Scholar]
  23. Osheroff N., Zechiedrich E. L. Calcium-promoted DNA cleavage by eukaryotic topoisomerase II: trapping the covalent enzyme-DNA complex in an active form. Biochemistry. 1987 Jul 14;26(14):4303–4309. doi: 10.1021/bi00388a018. [DOI] [PubMed] [Google Scholar]
  24. Osheroff N., Zechiedrich E. L., Gale K. C. Catalytic function of DNA topoisomerase II. Bioessays. 1991 Jun;13(6):269–273. doi: 10.1002/bies.950130603. [DOI] [PubMed] [Google Scholar]
  25. Reece R. J., Maxwell A. DNA gyrase: structure and function. Crit Rev Biochem Mol Biol. 1991;26(3-4):335–375. doi: 10.3109/10409239109114072. [DOI] [PubMed] [Google Scholar]
  26. Robinson M. J., Martin B. A., Gootz T. D., McGuirk P. R., Moynihan M., Sutcliffe J. A., Osheroff N. Effects of quinolone derivatives on eukaryotic topoisomerase II. A novel mechanism for enhancement of enzyme-mediated DNA cleavage. J Biol Chem. 1991 Aug 5;266(22):14585–14592. [PubMed] [Google Scholar]
  27. Robinson M. J., Martin B. A., Gootz T. D., McGuirk P. R., Osheroff N. Effects of novel fluoroquinolones on the catalytic activities of eukaryotic topoisomerase II: Influence of the C-8 fluorine group. Antimicrob Agents Chemother. 1992 Apr;36(4):751–756. doi: 10.1128/aac.36.4.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Robinson M. J., Osheroff N. Effects of antineoplastic drugs on the post-strand-passage DNA cleavage/religation equilibrium of topoisomerase II. Biochemistry. 1991 Feb 19;30(7):1807–1813. doi: 10.1021/bi00221a012. [DOI] [PubMed] [Google Scholar]
  29. Schneider E., Hsiang Y. H., Liu L. F. DNA topoisomerases as anticancer drug targets. Adv Pharmacol. 1990;21:149–183. doi: 10.1016/s1054-3589(08)60342-7. [DOI] [PubMed] [Google Scholar]
  30. Shen L. L., Mitscher L. A., Sharma P. N., O'Donnell T. J., Chu D. W., Cooper C. S., Rosen T., Pernet A. G. Mechanism of inhibition of DNA gyrase by quinolone antibacterials: a cooperative drug--DNA binding model. Biochemistry. 1989 May 2;28(9):3886–3894. doi: 10.1021/bi00435a039. [DOI] [PubMed] [Google Scholar]
  31. Sutcliffe J. A., Gootz T. D., Barrett J. F. Biochemical characteristics and physiological significance of major DNA topoisomerases. Antimicrob Agents Chemother. 1989 Dec;33(12):2027–2033. doi: 10.1128/aac.33.12.2027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Suto M. J., Domagala J. M., Roland G. E., Mailloux G. B., Cohen M. A. Fluoroquinolones: relationships between structural variations, mammalian cell cytotoxicity, and antimicrobial activity. J Med Chem. 1992 Dec 11;35(25):4745–4750. doi: 10.1021/jm00103a013. [DOI] [PubMed] [Google Scholar]
  33. Yamashita Y., Ashizawa T., Morimoto M., Hosomi J., Nakano H. Antitumor quinolones with mammalian topoisomerase II mediated DNA cleavage activity. Cancer Res. 1992 May 15;52(10):2818–2822. [PubMed] [Google Scholar]
  34. Yoshida H., Bogaki M., Nakamura M., Yamanaka L. M., Nakamura S. Quinolone resistance-determining region in the DNA gyrase gyrB gene of Escherichia coli. Antimicrob Agents Chemother. 1991 Aug;35(8):1647–1650. doi: 10.1128/aac.35.8.1647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zimmer C., Störl K., Störl J. Microbial DNA topoisomerases and their inhibition by antibiotics. J Basic Microbiol. 1990;30(3):209–224. doi: 10.1002/jobm.3620300312. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES