ktd Kocatepe Tıp Dergisi 3061-9904 Afyonkarahisar Sağlık Bilimleri Üniversitesi 10.18229/kocatepetip.883250 Clinical Sciences Klinik Tıp Bilimleri DNA BINDING ACTIVITIES OF THE VINCA ALKALOIDS AND PACLITAXEL AS ANTI-MICROTUBULE DRUGS USED IN CANCER THERAPY KANSER TEDAVİSİNDE KULLANILAN ANTİ-MİKROTÜBÜL İLAÇLARDAN VİNCA ALKALOİDLER VE PAKLİTAKSEL’İN DNA BAĞLANMA AKTİVİTELERİ https://orcid.org/0000-0003-4106-1056 Öksüzoğlu Emine AKSARAY UNIVERSITY 01 17 2022 23 1 51 57 02 19 2021 04 12 2021 Copyright © 1999, Kocatepe Tıp Dergisi 1999 Kocatepe Tıp Dergisi

OBJECTIVE: Microtubules are an essential part of the intracellular cytoskeletal structure and possess unique polymerization dynamics that are critical for many cellular functions, including cell division. Anti-microtubule drugs that interfere with microtubule formation are important chemotherapeutic agents for the treatment of various cancer. These drugs that block mitosis seem to work by a common mechanism, which suppresses the dynamics of microtubules, slows cells, induces apoptosis and subsequently kills tumor cells. Vinca alkaloids (vinblastine, vincristine and vinorelbine) and Taxanes (paclitaxel) are two different classes of anti-microtubule drugs that cause microtubule dysfunction and inhibit cancer cell proliferation. The main activity of vinca alkaloids and taxanes result from their binding interactions with tubulin proteins. However, studies on DNA interactions of these anti-microtubule drugs are not sufficient. In this study, it was aimed to investigate the DNA binding activities of the vinca alkaloids (vinblastine, vincristine, vinorelbine) and paclitaxel.MATERIAL AND METHODS: The interactions of the drugs with DNA were analyzed by agarose gel electrophoresis assay. Three types of DNA were used in each experiment, including 100bp marker DNA, pUC19 plasmid DNA (2686 bp), and pBR322 plasmid DNA (4361 bp). After the DNAs were incubated with different concentrations of the drugs under certain conditions, agarose gel electrophoresis was performed. DNA band distributions were analyzed with a gel analysis system so that the drugs-DNA interactions could be interpreted.RESULTS: According to our results, it was found that among the vinca alkaloids, especially vinorelbine binds to DNA with higher activity than vincristine and vinblastine. The Vinca alkaloids have structural properties required for DNA binding activity and there is a similarity in their DNA binding models. However, the results showed that paclitaxel, which is from the taxane group, did not have DNA binding activity. This may be because the chemical structure of paclitaxel is not suitable for binding to DNA.CONCLUSIONS: The interaction of drugs with DNA play an important role in determining the pathways of drugs action and their ability to cause DNA damage. Consequently, the findings of our study will contribute to elucidating the effect mechanisms and the genotoxic potentials of these drugs, which are microtubule inhibitors.

AMAÇ: Mikrotübüller, hücre içi hücre iskeleti yapısının önemli bir parçasıdır ve hücre bölünmesi dahil birçok hücresel işlev için kritik olan benzersiz polimerizasyon dinamiklerine sahiptirler. Mikrotübüllere müdahale eden anti-mikrotübül ilaçlar, çeşitli kanserlerin tedavisi için çok önemli kemoterapötik ajanlardır. Mitozu bloke eden bu ilaçlar, mikrotübüllerin dinamiğini baskılayan, hücreleri yavaşlatan, apoptozu indükleyen ve ardından tümör hücrelerini öldüren ortak bir mekanizma ile çalışmaktadır. Vinka alkaloidleri (vinblastin, vincristin ve vinorelbin) ve Taksanlar (paklitaksel), mikrotübüllerin işlev bozukluğuna neden olan ve kanser hücresi proliferasyonunu inhibe eden iki farklı anti-mikrotübül ilaç sınıfıdır. Vinka alkaloidlerinin ve taksanların ana aktivitesi, tübülin proteinleri ile bağlanma etkileşimlerinden kaynaklanmaktadır. Ancak, bu anti-mikrotübül ilaçların DNA etkileşimleri ile ilgili çalışmalar yeterli değildir. Bu çalışmada, vinka alkaloidleri (vinblastin, vincristin, vinorelbin) ve paklitakselin DNA bağlanma aktivitelerinin araştırılması amaçlanmıştır.GEREÇ VE YÖNTEM: İlaçların DNA ile etkileşimleri agaroz jel elektroforez deneyi ile analiz edildi. Her deneyde, 100 bp marker DNA, pUC19 plazmid DNA (2686 bp) ve pBR322 plazmid DNA (4361 bp) dahil olmak üzere üç tip DNA kullanıldı. DNA'lar belirli koşullar altında farklı ilaç konsantrasyonları ile inkübe edildikten sonra agaroz jel elektroforezi yapıldı. İlaç-DNA etkileşimlerinin yorumlanabilmesi için DNA bant dağılımları jel analiz sistemi ile analiz edildi.BULGULAR: Sonuçlarımıza göre vinka alkaloidlerinden özellikle vinorelbinin, vinkristin ve vinblastine göre daha yüksek aktiviteyle DNA'ya bağlandığı bulunmuştur. Vinca alkaloidleri, DNA bağlanma aktivitesi için gerekli yapısal özelliklere sahiptir ve DNA bağlanma motiflerinde benzerlik vardır. Ancak sonuçlar, taksan grubundan olan paklitakselin DNA bağlama aktivitesine sahip olmadığını gösterdi. Bunun nedeni, paklitakselin kimyasal yapısının DNA'ya bağlanmaya uygun olmaması olabilir.SONUÇ: İlaçların DNA ile interaksiyonu, ilaçların etki yollarını ve DNA hasarına neden olma yeteneklerini belirlemede önemli bir rol oynar. Sonuç olarak, çalışmamızın bulguları mikrotübül inhibitörü olan bu ilaçların etki mekanizmalarını ve genotoksik potansiyellerini aydınlatmaya katkı sağlayacaktır.

 Anti-microtubule drugs Vinca Alkaloids Paclitaxel DNA-binding Cancer Therapy Anti-mikrotübül ilaçlar Vinca alkaloidleri Paklitaksel DNA-bağlanma Kanser terapi 1. Daniel R, Bumdan, Alan H. Calvert and Rowinski, EK, eds. Handbook of Anticancer Drug Development. Lippincott Williams & Wilkins, Baltimore, Maryland, USA, 2003. 2. Pommier Y, Yu Q and Kohn KW. Chapter 2: Novel Targets in the Cell Cycle and Cell Cycle Checkpoints, In: Baguley, BC and Kerr DJ, eds. Anticancer Drug Development, Elsevier Inc. 2002;3-30. 3.Fischer, PM. Chapter 11: Cell Cycle Inhibitors in Cancer:Current Status and Future Directions. In: Neidle S. ed. Cancer Drug Design and Discovery, Elsevier Inc. 2008;253-83. 4. Boyer, MJ and Tannock, IF. Chapter 17: Cellular and Molecular Basis of Drug Treatment for Cancer, In: Tannock, IF, Hill, RP. Bristow RG and Harrington, L. The Basic Science of Oncology, 4th Edition, The McGraw-Hill Companies, Inc. 2005;349-75. 5.Schummel PH, Gao M, Winter R. Modulation of thePolymerization Kinetics of alpha/beta-Tubulin by Osmolytes and Macromolecular Crowding. Chemphyschem. 2017;18:189- 97. 6.Waterman-Storer CM, Salmon ED. Microtubule dynamics: treadmilling comes around again. Curr Biol.1997;7(6):369-72. 7.Avendaño, C. and Menéndez, JC. Medicinal Chemistryof Anticancer Drugs. Chapter 8: Anticancer Drugs Targeting Tubulin and Microtubules, Elsevier B. V. 2008;229-49. 8.Checchi PM, Nettles JH, Zhou J, Snyder JP, Joshi HC.Microtubule-interacting drugs for cancer treatment. Trends Pharmacol Sci. 2003;24:361–65. 9.Jordan MA, Wilson L. Microtubules as a target for anticancer drugs. Nat Rev Cancer. 2004;4:253–65. 10.Zhou J, Giannakakou P. Targeting microtubulesfor cancer chemotherapy. Curr Med Chem Anticancer Agents. 2005;5(1): 65-71. 11.Jordan MA, Kamath K. How do microtubule-targeted drugs work? An overview. Curr Cancer Drug Targets 2007;7(8):730-42. 12.Jordan MA. Mechanism of action of antitumor drugs that interact with microtubules and tubulin. Curr Med Chem Anticancer Agents 2002;2:1-17. 13. Estève MA, Carré M, Braguer D. Microtubules in apoptosis induction: are they necessary? Curr Med Chem Anticancer Agents. 2003;3(4): 291-306. 14.Beck W, Cass CE, Houghton PJ. Microtubule-targetinganticancer drugs derived from plants and microbes: vinca alkaloids, taxanes and epothilones. In: Cancer Medicine, 5th edition. Bast, Kufe, Pollock, Weichselbaum, Holland, Frei. B. C. Decker, 2000. 15. Bates D, Eastman A. Microtubule destabilising agents: far more than just antimitotic anticancer drugs. Br J Clin Pharmacol. 2017;83:255-68. 16.Mukhtar E, Adhami VM, Mukhtar H. Targeting Microtubules by Natural Agents for Cancer Therapy. Mol Cancer Ther. 2014;13(2): 275–84. 17.Moudi MR, Go R, Yien CY, Nazre M. Vinca alkaloids. IntJ Prev Med. 2013;4(11):1231-5. 18. Rowinsky EK, Onetto N, Canetta RM, Arbuck SG. Taxol: the first of the taxanes, an important new class of antitumor agents. Semin Oncol. 1992;19:646-62. 19.Abal M, Andreu JM, Barasoain I. Taxanes: microtubule and centrosome targets, and cell cycle dependent mechanisms of action. Curr Cancer Drug Targets. 2007;7(8):713- 29. 20.Rowinsky E. The Vinca Alkaloids. In: Kufe DW, PollockRE, Weichselbaum RR, et al., editors. Holland-Frei Cancer Medicine. 6th edition. Hamilton (ON): BC Decker; 2003. 21.Risinger AL, Giles FJ, Mooberry SL. Microtubule dynamics as a target in oncology. Cancer Treat Rev. 2009;(3): 255-61. 22. Weaver BA. How Taxol/paclitaxel kills cancer cells. Mol Biol Cell. 2014;25(18):2677-81. 23.Pandya P, Gupta SP, Pandav K, Barthwal R, Jayaram B,Kumar S. DNA binding studies of Vinca alkaloids: experimental and computational evidence. Nat Prod Commun. 2012;7(3): 305-9. 24.Mohammadgholi A, Rabbani-Chadegani A, Fallah S.Mechanism of the interaction of plant alkaloid vincristine with DNA and chromatin: spectroscopic study. DNA Cell Biol. 2013;32(5):228-35. 25. Bischoff G, Hoffmann S. DNA-binding of drugs used in medicinal therapies. Curr Med Chem. 2002; 9(3): 312-48. 26.Weber GF. DNA Damaging Drugs. Molecular Therapies of Cancer. 2014; 9-112. 27.Chen JG, Yang CP, Cammer M, Horwitz SB. Gene expression and mitotic exit induced by microtubule-stabilizing drugs. Cancer Res. 2003;63:7891–9. 28.Selvaraj C, Singh SK. Computational and Experimental Binding Mechanism of DNA-drug Interactions. Curr Pharm Des. 2018;24(32): 3739-57.