本實驗室所研發的3,6-bis(1-methyl-4-vinylpyridinium) carbazole diiodide (BMVC)是一個良好的四股結構穩定者,在細胞實驗下顯示其能有效抑制癌症細胞增生,而且與DNA作用後,其螢光性質會增強,所以亦可作為一個相當優良的螢光探針,為了研發更強而有利的四股結構穩定者,我們想要了解BMVC與DNA的作用模式為何?與不同結構的DNA的作用差異又是如何?所以我們利用拉曼光譜能以振動態角度去觀察兩物質作用機制的優勢,我們希望能由此得到更多有利的研發訊息,但由於螢光對於拉曼光譜而言是相當困擾的一件事,所以我們改採用結構類似但沒有螢光性質的3,6-bis(1-methyl -3- vinylpyrazinium) carbazole diiodide (BMVC-4),期望從分析其與DNA作用後的拉曼光譜差異,讓我們可以去模擬BMVC是如何與不同結構的DNA作用,並從中獲得有利訊息去研發更具潛力的新四股結構穩定者!

Since last few decades, it is known that inhibition of telomerase ability is a key to suppress cancer cells proliferation. In the human telomeres, there is about 150-200bs single strand with (TTAGGG)n template repeats. Of particular interest is that the G-rich sequences can form G-quadruplex structures in the presence of monovalent cations such as Na+ and K+. Therefore, it is possible to stabilize the G-quadruplex for reducing the telomerase ability and then suppressing the cancer cells proliferation. Our laboratory have developed a novel G-quadruplex stabilizer, 3,6-bis(1-methyl-4-vinyl pyridinium) carbazole diiodide (BMVC), which can inhibit telomerase activity. In order to design a better G-quadruplex stabilizer, we intended to determine the binding mode of BMVC. Raman spectroscopy has the potential to investigate the detail binding mode at point of view from vibration motions. However the fluorescence of BMVC can be a serious problem upon interact with G-quadruplex. Here we choose 3,6-bis (1-methyl-3-vinyl pyrazinium) carbazole diiodide (BMVC-4)to do Raman experiment which is structural analogous to BMVC. BMVC-4 has negligible fluorescence interference even upon interact with DNA. Our experiment results allow us to investigage how BMVC-4 interacts with G-quadruplex, and of course, the binding mode. We believe that the realization of BMVC-4/G-quadruplex binding mode can provide important information for designing a better drug to suppress cancer cell proliferation.

(1) Telomeres, ed. E. H. Blackburn and C. W. Greider, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1996.
(2) E. H. Blackburn and J. W. Szostake, Annu. Rev. Biochem., 1984,53, 163.
(3) J. R. Williamson, Annu. Rev. Biophys. Biomol. Struct., 1994, 23,703.
(4) Harley, C. B.; Futcher, A. B.; Greider, C. W. Nature 1990, 345, 458-460.
(5) Lundblad, V.; Szostak, J. W. Cell 1989, 57, 633-643.
(6) Sandell, L. L.; Zakian, V. A. Cell 1993, 75, 729-739.
(7) Harley, C. B.; Villeponteau, M. P. Curr. Opin. Genet. Dev. 1995, 5, 249-
255.
(8) Greider, C. W.; Blackburn, E. H. Cell 1987, 51, 887-898.
(9) Feng, J.; Funk, W. D.; Wang, S.-S.; Weinrich, S. L.; Avilion, A. A.; Chiu,C.-P.; Adams, R. R.; Chang, E.; Allsopp, R. C.; Yu, J.; Le, S.; West, M. D.;Harley, C. B.; Andrews, W. H.; Greider, C. W.; Villeponteau, B. Science
1995, 269, 1236-1241.
(10) Hahn, W. C.; Counter, C. M.; Lundberg, A. S.; Beijersbergen, R. L.; Brooks,M. W.; Weinberg, R. A. Nature 1999, 400, 464-468.
(11) Kim, N. W.; Piatyszek, M. A.; Prowas, K. R.; Harley, C. B.; West, M. D.; Ho,P. L.; Coviello, G. M.; Wright, W. E.; Weinrich, S. L.; Shay, J. W. Science 1994, 266, 2011-2015.
(12) Blackburn, E. H. Nature 1991, 350, 569-573.
(13) Breslow, R. A.; Shay, J. W.; Gazdar, A. F.; Srivastava, S. J. Natl. Cancer Inst.1997, 89, 618-623.
(14) Mergny, J. L.; He´ le`ne, C. Nat. Med. 1998, 4, 1366-1367.
(15) Han, H.; Hurley, L. H. Trends Pharmacol. Sci. 2001, 21, 136-142.
(16) Gellert, M.; Lipsett, M. N.; Davies, D. R. Proc. Natl. Acad. Sci. U.S.A. 1962, 48, 2013-2018.
(17) Arthanari, H.; Basu, S.; Kawano, T. L.; Bolton, P. H. Nucleic Acids Res. 1998,26, 3724-3728
(18) Zahler, A. M.; Williamson, J, R.; Cech, T. R.; Prescott, D. M. Nature 1991, 350, 718-720.
(19) Neidle, S.; Parkinson, G. N. Nat. Rev. Drug Discovery 2003, 1,383-393.
(20) Kerwin, S. M. Curr. Pharm. Des. 2000, 6, 441-478.
(21) Chang, C. C.; Wu, J. Y.; Chang, T.-C. J. Chin. Chem. Soc. 2003,50, 185-188.
(22) Chang, C. C.; Wu, J. Y.; Chien, C. W.; Wu, W. S.; Liu, H.; Kang,C. C.; Yu, L. J.; Chang, T.-C. Anal. Chem. 2003, 75, 6177-6183.
(23) Chang, C. C.; Kuo, I.-C.; Ling, I.-F.; Chen, C. T.; Chen, H. C.;Lou, P. J.; Lin, J. J.; Chang, T.-C. Anal. Chem. 2004, 76, 4490-4494.
(24) Chang, C. C.; Chu, J. F.; Kao, F. J.; Chiu, Y. C.; Lou, P. J.; Chen,
H. C.; Chang, T.-C. Anal. Chem. 2006, 78, 2810-2815.
(25) Kang, C.-C.; Chang, C.-C.; Cheng, J.-Y.; Chang, T.-C. J. Chin.Chem. Soc. 2005, 52, 1069-1072.
(26) Chang, C. C.; Chu, J. F.; Kuo, H.H.; Kang, C.-C.; Lin, S. H.; Chang, T. C. J. Lumin. 2006, 119-120, 84-90.
(27) Wang, Y.; Patel, D. J. Structure 1993, 1, 262-283.
(28) Parkinson, G. N.; Lee, M. P. H.; Neidle, S. Nature 2002, 417, 876-880.
(29) Ambrus,A., Chen,D., Dai,J., Bialis,T., Jones,R.A. and Yang,D. Nucleic Acids Res., 2006,34, 2723–2735.
(30) Rujan,I.N., Meleney,J.C. and Bolton,P.H., Nucleic Acids Res., 2005,33, 2022–2031.
(31) Li,J., Correia,J.J., Wang,L., Trent,J.O. and Chaires,J.B., Nucleic Acids Res., 2005, 33, 4649–4659.
(32) Redon,S., Bombard,S., Elizondo-Riojas,M.A. and Chottard,J.C.,Nucleic Acids Res., 2003, 31,1605–1613.
(33) He,Y., Neumann,R.D. and Panyutin,I.G., Nucleic Acids Res., 2004,32, 5339–5367.
(34) Xu,Y., Noguchi,Y. and Sugiyama,H., Bioorg. Med. Chem., 2006, 14, 5584–5591.
(35) Phan,A.T. and Patel,D.J., J. Am.Chem. Soc., 2003, 125, 15021–15027.
(36) Daekyu Sun, Wei-Jun Liu, Kexiao Guo, Jadrian J. Rusche, Scot Ebbinghaus, Vijay Gokhale, and Laurence H. Hurley MolCancer Ther 2008; 7(4), 880-889.
(37) Lundblad, V., Szostak, J. W. Cell 1989, 57, 633-643.
(38) Sandell, L. L., Zakian, V. A., Cell 1993, 75, 729-739.
(39) Harley, C. B., Villeponteau, M. P. Curr. Opin. Genet. DeV. 1995, 5, 249-255.
(40) Meyne, J., Ratliff, R. L.; Moyzis, R. K. Proc. Natl. Acad. Sci. U.S.A. 1989, 86, 7049-7053.
(41) Morin, G. B. Cell 1989, 59, 521-529.
(42) Sullivan DC, Bicknell R., J Cancer 2003;89:228–31.
(43) Giordano FJ, Johnson RS., Curr Opin Genet Dev 2001;11:35–40.
(44) Goodsell DS., Stem Cells 2003;21:118–9.
(45) Jain RK., Semin Oncol 2002;29:3–9.
(46) Loureiro RM, D’Amore PA., Cytokine Growth Factor Rev 2005;16: 77–89.
(47) Finkenzeller G, Sparacio A, Technau A., Oncogene 1997;15:669–76.
(48) Siddiqui-Jain A, Grand CL, Bearss DJ, Hurley LH., Proc Natl Acad Sci U S A 2002;99: 11593–8.
(49) De Armond R, Wood S, Sun D, et al., Biochemistry 2005;44:16341–50.
(50) Sun D, Guo K, Rusche JJ, et al., Nucleic Acids Res 2005;33:6070–80.
(51) Dexheimer TS, Sun D, Hurley LH., J Am Chem Soc 2006;128:5404–15.
(52) Cogoi S, Xodo LE., Nucleic Acids Res 2006;34:2536–49.
(53) Shirude PS, Okumus B, Ying L, et al., J Am Chem Soc 2007; 129: 7484–5.
(54) Qin Y, Rezler EM, Gokhale V, et al., Nucleic Acids Res 2007 ; 35: 7698–713.
(55) Guo K, Pourpak A, Beetz-Rogers K, et al., J Am Chem Soc 2007; 129: 10220–8.
(56) C. V. Raman and K. S. Krishnan, Nature, 121(3048), 501, March 31, 1928
(57) Alan Campion and Patanjali Kambhampati, Chemical Society Reviews, 1998, volume 27
(58) Martin Moskovits, J. Raman Spectrosc. 2005; 36: 485–496
(59) Electromagnetic Mechanism of Surface-enhanced Spectroscopy,George C. Schatz and Richard P. Van Duyne
(60) B. N. J. Persson,1,2,3 Ke Zhao,4 and Zhenyu Zhang1, PRL 2006；96, 207401
(61) Nicolae Leopold†,‡ and Bernhard Lendl*,‡,J. Phys. Chem. B 2003, 107, 5723-5727
(62) A. Bree,R. Zwarich. J. Chem. Phys 1968 , 49,3344-3355
(63) Debra S. Egolf,‡ Mark R. Waterland,§ and Anne Myers Kelley*§, J. Phys. Chem. B 2000, 104, 10727-10737
(64) R. Anandhi and S. Umapathy*,J. Raman Spectrosc. ,1998,29, 901-906
(65) M. P. M. Marques,1,2∗ P. J. Oliveira,3 A. J. M. Moreno3 and L. A. E. Batista de Carvalho1, J. Raman Spectrosc. 2002; 33: 778–783
(66) Juan Francisco Arenas, Mark Steven Woolley, Juan Carlos Otero,* and Juan Ignacio Marcos, J. Phys. Chem. 1996, 100, 3199-3206
(67) Juan F. Arenas, Mark S. Woolley, Isabel Lo´pez Toco´ n, Juan C. Otero(a) and Juan I. Marcos(b) , J.Chem.Phys. 2000,112,7669-7683
(68) Gerhard Stock, Clemens Woywod, and Wolfgang Domcke, Tim Swinneya) and Bruce S. Hudson , J.CHem.Phys.1995,103,6851-6860
(69) Henrietta Endre´dia, Ferenc Billesa,*, Ga´bor Kereszturyb , Journal of Molecular Structure (Theochem) , 2004 , 677,211–225
(70) J.F. Arenas, S.P. Centeno, I. Lo´pez-Toco´n, J.C. Otero*,Vibrational Spectroscopy ,2004,35, 39–44
(71) J.F. Arenas, I. Lo´pez Toco´n, J.C. Otero, J.I. Marcos ), Vibrational Spectroscopy ,1999,19,213–221