Loading...
|
Please use this identifier to cite or link to this item:
http://libir.tmu.edu.tw/handle/987654321/36369
|
| Title: | 以天然物為cap與氮-羥基肉桂酸醯胺結合作為組蛋白去乙醯酶抑制劑 |
| Authors: | 趙世偉 |
| Contributors: | 生藥學研究所 |
| Date: | 2010 |
| Issue Date: | 2010-10-20 12:42:21 (UTC+8) |
| Abstract: | 研究顯示，組蛋白去乙醯酶抑制劑可望發展成為抗癌藥物，所有已知之組蛋白去乙醯酶抑制劑 (HDACi)，從化學結構上包含了三個藥理活性基團：與鋅產生螯合之基團，疏水性鏈長，與疏水性的辨認部位。在本研究我們以具有疏水性之中藥成份蛇床子素 (osthole)，來當作疏水性的辨認部位，合成一系列以N-hydroxycinnamides為基本架構之組蛋白去乙醯酶抑制劑 (HDACi)，以探討蛇床子素 (osthole) 對於活性的效果。我們合成了九個結合蛇床子素 (osthole) 之N-hydroxacinnamides (9a~9i)，並同時進行酵素抑制活性測試，發現化合物9d, 9e, 9g 具有顯著抑制子宮頸癌細胞萃取之組蛋白去乙醯酶 (HeLa cell nuclear extract HDAC) (IC50: 24.5, 20.0, 19.6 nM)，其抑制活性相當於目前臨床上用於治療表皮T細胞淋胞癌 (cutaneous T-cell lymphoma, CTCL) 之組蛋白去乙醯酶抑制劑 (HDACi) suberoylanilide hydroxamic acid (SAHA, IC50: 24.5 nM)，另外在組蛋白去乙醯酶1型 (HDAC1) 和組蛋白去乙醯酶6型 (HDAC6) 之酵素抑制活性上，化合物9d及9e具有和SAHA類似之活性，9g則對組蛋白去乙醯酶抑制劑1型 (HDAC1) 具有較好的選擇性，從乙醯化組蛋白3型 (acetylated H3) 與乙醯化α-微管蛋白 (acetylated α-tubulin) 之促進實驗中，化合物9d, 9e, 9g與SAHA相比較，9d有明顯之促進乙醯化之作用，顯示其具有好之細胞效果，接著將9d與SAHA進行分子結合模擬分析，顯示9d之蛇床子素 (osthole) 基團與SAHA的苯環都是作用酵素口袋表面疏水區，且蛇床子素 (osthole) 之側鏈經過修飾，可能產生對第Ⅱa型組蛋白去乙醯酶 (class Ⅱa HDAC) 選擇性抑制，以上這些結果證實，以蛇床子素 (osthole) 為疏水性cap與N-hydroxacinnamides相結合，可發展出有效之組蛋白去乙醯酶抑制劑 (HDACi)。
Histone deacetylase (HDAC) inhibitors are regarded as promising therapeutics for the treatment of cancer. All reported HDAC inhibitors contain three pharmacophoric features: a zinc-chelating group, a hydrophobic linker, and a hydrophobic cap for surface recognition. In this study we investigated the effectiveness of osthole, a hydrophobic Chinese herbal compound, as the surface recognition cap in hydroxamate-based compounds as inhibitors of HDAC. Nine novel osthole-based N-hydroxycinnamides were synthesized and screened for enzyme inhibition activity. Compounds 9d, 9e, 9g exhibited inhibitory activities (IC50=24.5, 20.0, 19.6 nM) against nuclear HDACs in HeLa cells comparable to that of suberoylanilide hydroxamic acid (SAHA; IC50=24.5 nM), a potent inhibitor clinically used for the treatment of cutaneous T-cell lymphoma (CTCL). While compounds 9d and 9e showed SAHA-like activity towards HDAC1 and HDAC6, compound 9g was more selective for HDAC1. Compound 9d exhibited the best cellular effect, which was comparable to that of SAHA, of enhancing acetylation of either α-tubulin or histone H3. Molecular docking analysis showed that the osthole moiety of compound 9d may interact with the same hydrophobic surface pocket exploited by SAHA and it may be modified to provide class-specific selectivity. These results suggest that osthole is an effective hydrophobic cap when incorporated into N-hydroxycinnamide-derived HDAC inhibitors. |
| Relation: | 137頁 |
| Description: | 中文摘要 i
Abstract iii
表目錄 vii
流程圖目錄 viii
圖目錄 ix
一、緒論及研究目的 - 1 -
二、結果與討論 - 9 -
1. 化學合成 - 9 -
2. 化合物 (9a-i) 對於抑制子宮頸癌細胞萃取之組蛋白去乙醯酶(HeLa cell nuclear extract histone deacetylase) 與抑制肝癌細胞 (HepG2) 生長之活性 - 12 -
3. 化合物9d, 9e, 9g, 9i對組蛋白去乙醯酶1、6型 (HDAC1，-6)
酵素抑制活性 - 14 -
4. 化合物9d, 9e, 9g對於肝癌細胞HepG2α-微管蛋白乙醯化
(acetylation α-tubulin) 及組蛋白三型乙醯化 (acetylation
histone 3) 之影響 - 15 -
5. 化合物9d之分子模擬(molecular modeling) - 17 -
6. 化合物 9d 對於正常細胞之細胞毒性 - 19 -
三、結論 - 20 -
四、實驗部份 - 21 -
I. 儀器與實驗材料 - 21 -
1. 一般儀器及方法 - 21 -
II. 化學合成步驟及物理資料 - 24 -
III. 生物活性測試方法 - 49 -
1. HDAC activity assay - 49 -
2. The sulforhodamine B (SRB) growth inhibition assay - 50 -
3. Western blot analysis - 51 -
五、參考文獻(References) - 52 -
六、附圖 - 60 -
1.Matsudaira, P.T.L.; Harvey, F; Arnold, B.; Kaiser, C.;
Monty, K.; Matthew, P.; Anthony, B.; Hidde, P., Molecular
Cell Biology, 2007.
2.Alberts, B., Molecular biology of the Cell. 2002, New
York.
3.Jemal, A.; Murray, T.; Ward, E.; Samuels, A.; Tiwari,
R.C.; Ghafoor, A.; Feuer, E.J.; Thun, M.J., CA Cancer J.
Clin., 2005, 55, 10.
4.Harper, D.M.; Franco, E.L.; Wheeler, C.; Ferris, D.G.;
Jenkins, D.; Schuind, A.; Zahaf, T.; Innis, B.; Naud, P.;
De Carvalho, N.S.; Roteli-Martins, C.M.; Teixeira, J.;
Blatter, M.M.; Korn, A.P.; Quint, W.; Dubin, G., Lancet,
2004, 364, 1757.
5.David, M.K.; Anna, C., Nature Reviews Microbiology, 2008,
6, 211.
6.Luger, K.; Mader, A.W.; Richmond, R.K.; Sargent, D.F.;
Richmond, T.J., Nature, 1997, 389, 251.
7.Davie, J.R., Curr Opin Genet Dev, 1998. 8, 173.
8.Cress, W.D. ; Seto, E.; J Cell Physiol, 2000, 184, 1.
9.Munshi, A.; Shafi, G.; Aliya. N.; Jyothy A., J. Genet.
Genomics, 2009, 36, 75.
10.Takayoshi, S.; Naoki, M., Curr. Med. Chem., 2006, 13,
935.
11.Herman, J.G.; Baylin, S.B., New. Engl. J. Med., 2004,
350, 947.
12.Roth, S.Y.; Denu, J.M.; Allis, C.D., Annu. Rev.
Biochem., 2001, 70, 81.
13.David, E.S.; Shelley, L.B., Microbiol. Mol. Biol. Rev.,
2000, 64, 435.
14.Hassig, C.A.; Schreiber, S.L., Curr. Opin. Chem. Biol,
1997, 1, 300.
15.Gregoire, S.; Yang, X.J., Mol. Cell. Biol., 2005, 25,
2873.
16.Khochbin, S.; Verdel, A.; Lemercier, C.; Seigneurin-
Berny, D., Curr. Opin. Genet. Dev., 2001, 11, 162.
17.Annemieke, J.M.; Ruijter, D.; Albert, H.; GENNIP, V.;
CARON, H.N.; Kemp, S.; Andre, B.P.; Kuilenburg V.,
Biochem. J., 2003, 370, 737.
18.Gregoretti, I.V.; Lee, Y.M.; Goodson, H.V., J. Mol.
Biol, 2004, 338, 17.
19.Robertson, K.D.; Wolffe, A.P., Nature Rev. Genet., 2000,
1, 11.
20.Feng, Q.; Zhang, Y., Genes & Dev., 2001, 15, 827.
21.Fuks, F.B.; Wendy, A.; Godin, N.; Kasai, M.; Kouzarides,
T., EMBO J., 2001, 20, 2536.
22.Verdin, E.; Dequiedt, F.; Kasler, H.G., Trends Genet.,
2003, 19, 286.
23.Mckinsey, T.A.; Zhang, C.L.; Lu, J.; Olson, E. N.,
Nature, 2000, 408, 106.
24.Wade, P.A., Hum Mol Genet, 2001, 10, 693.
25.Hubbert, C.; Guardiola, A.; Shao, R.; Kawaguchi, Y.;
Ito, A.; Nixon, A.; Yoshida, M.; Wang, X.F.; Yao, T.P.,
Nature, 2002, 417, 455.
26.North, B.J.; Marshall, B.L.; Borra, M.T.; Denu, J.M.;
Verdin, E., Mol. Cell, 2003, 11, 437.
27.Vigushin, D.M.; Coombes, R.C., Anti-Cancer Drugs, 2002,
13, 1.
28.Olaf, W. ; Hedwig, E.D.; Till, M.; Ina, O., Cancer
Lett., 2009, 277, 8.
29.Abraham, J., Community Oncology, 2007, 4, 384.
30.Glaser, K.B.; Li, J.; Staver, M.J.; Wei, R.Q.; Albert,
D.H.; Davidsen, S.K., Biochem Biophys Res Commun, 2003,
310, 529.
31.Paris, M.; Porcelloni, M.; Binaschi, M.; Fattori, D., J
Med Chem, 2008, 51, 1505.
32.Yoshida, M.; Kijima, M.; Akita, M.; Beppu, T., J Biol
Chem, 1990, 265, 17174.
33.Han, J.W.; Ahn, S.H.; Park, S.H.; Wang, S.Y.; Bae, G.U.;
Seo, D.W.; Kwon, H. K.; Hong, S.; Lee, H.Y.; Lee, Y.W.;
Lee, H.W., Cancer Res, 2000, 60, 6068.
34.Yoshida, M.; Horinouchi, S.; Beppu, T., BioEssays, 1995,
17, 423.
35.Closse, A.; H. R., Helv. Chim. Acta, 1974, 57, 553.
36.Kawai, M.; Rich, D.H.; Walton, J.D., Biochem. Biophys.
Res. Comm., 1983, 111, 398.
37.Itazaki, H.; Nagashima, K.; Sugita, K.; Yoshida, H.;
Kawamura, Y.; Yasuda, Y.; Matsumoto, K.; Ishii, K.;
Uotani, N.; Nakai, H.; Terui, A.; Yoshimatsu, S.;
Ikenishi, Y.; Nakagawa, Y., J. Antibiot., 1990, 63, 1524.
38.Furumai, R.; Matsuyama, A.; Kobashi, N.; Lee, K.H.;
Nishiyama, M.; Nakajima, H.; Nakajima, A.; Komatsu, Y.;
Nishino, N.; Yoshida, M.; Horinouchi, S., Cancer Res.,
2002. 62, 4916.
39.Fournel, M.; Trachy-Bourget, M.C.; Theresa Y.P.; Kalita,
A., Cancer Res., 2002, 62, 4325.
40.Richon, V.M.; Webb, Y.; Merger, R.; Sheppard, T.;
Jursic, B.; Ngo, L.; Civoli, F.; Breslow, R.; Rifkind,
R.A.; Marks, P.A., Proc. Natl. Acad. Sci. U.S.A., 1996,
93, 5705.
41.Richon, V.M.; Emiliani, S.; Verdin, E.; Webb, Y.;
Breslow, R.; Rifkind R.A.; Paul, A.M., Proc. Natl. Acad.
Sci. U.S.A., 1998, 95, 3003.
42.Cohen, L.A.; Amin, S.; Marks, P.A.; Rifkind, R.A.;
Desai, D.; Richon, V.M., Anticancer Res., 1999, 19, 4999.
43.Wittich, S.; Scherf, H.; Xie, C.; Brosch, G.; Loidl, P.;
Gerhauser, C.; Jung, M., J. Med. Chem., 2002, 45, 3296.
44.Furumai, R.; Komatsu, Y.; Nishino, N.; Khochbin, S.;
Yoshida, M.; Horinouchi, S., Proc. Natl. Acad. Sci.
U.S.A., 2001, 98, 87.
45.Komatsu, Y.; Tomizaki, K.Y.; Tsukamoto, M.; Kato, T.;
Nishino, N.; Sato, S.; Sato, T.; Tsuruo, T.; Furumai,
R.; Yoshida, M.; Horinouchi, S.; Hayashi, H., Cancer
Res., 2001, 61, 4459.
46.Plumb, J.A.; Finn, P.W.; Williams, R.J.; Bandara, M.J.;
Romero, M.R.; Watkins, C.J.; Thangue, N.B.; Brown, R.,
Mol. Cancer. Ther., 2003, 2, 721.
47.Remiszewski, S.W., Curr. Med. Chem., 2003, 10, 2393.
48.Remiszewski, S.W.; Sambucetti, L.C.; Bair, K.W.;
Bontempo, J., J. Med. Chem., 2003, 46, 4609.
49.Atadja, P.; Gao, L.; Kwon, P.; Trogani, N.; Walker, H.;
Hsu, M., Cancer Res., 2004, 64, 689.
50.Bouchain, G.; Leit, S.; Frechette, S.; Khalil, E.A.;
Lavoie, R., J. Med. Chem., 2003, 46, 820.
51.Besterman, J.M., Pacifichem, 2005.
52.Ueda, H.; Nakajima, H.; Hori, Y.; Fujita, T.; Nishimura,
M.; Goto, T.; Okuhara, M., J Antibiot., 1994, 47, 301.
53.Richon, V.M.; Emiliani, S.; Verdin, E.; Webb, Y.;
Breslow, R.; Rifkind, R.A.; Marks, P. A., Proc Natl Acad
Sci U S A 1998, 95, 3003.
54.Monks, A.; Hose, C.D.; Pezzoli, P.; Kondapaka, S.;
Vansant, G.; Petersen, K. D.; Sehested, M.; Monforte,
J.; Shoemaker, R.H., Anticancer Drugs, 2009, 20, 682.
55.Wang, H.; Yu, N.; Song, H.; Chen, D.; Zou, Y.; Deng, W.;
Lye, P.L.; Chang, J.; Ng, M.; Sun, E.T.; Sangthongpitag,
K.; Wang, X.; Wu, X.; Khng, H.H.; Fang, L.; Goh, S.K.;
Ong, W.C.; Bonday, Z.; Stunkel, W.; Poulsen, A.;
Entzeroth, M., Bioorg. Med. Chem. Lett., 2009, 19, 1403.
56.Bottomley, M.J.; Lo, S. P.; Di Giovine, P.; Cirillo, A.;
Scarpelli, R.; Ferrigno, F.; Jones, P.; Neddermann, P.;
De Francesco, R.; Steinkuhler, C.; Gallinari, P.; Carfi,
A., J. Biol. Chem., 2008, 283, 26694.
57.Nielsen, T.K.; Hildmann, C.; Dickmanns, A.;
Schwienhorst, A.; Ficner, R., J. Mol. Biol, 2005, 354,
107.
58.Taunton, J.; Hassig, C.A.; Schreiber, S.L., Science,
1996, 272, 408.
59.Vaisburg, A.; Bernstein, N.; Frechette, S.; Allan, M.;
Abou-Khalil E., Bioorg. Med. Chem. Lett., 2004, 14, 283.
60.Bouchain, G.; Delorme, D., Curr. Med. Chem., 2003, 10,
2359.
61.Woo, S.H.; Frechette, S.; Khalil, E.A.; Bouchain, G.;
Vaisburg, A.; Moradei, O.; Bernstein, N.; Leit, S.;
Allan, M.; Fournel, M.; Trachy-Bourget, Z.; Besterman,
J. M.; Delorme, D., J. Med. Chem., 2002, 45, 2877.
62.Frey, R.R.; Wada, C.K.; Garland, R.B.; Curtin, M.L.;
Michaelides, R.L.; Michaelides, J.; Pease, L.J.; Glaser,
K.B.; Marcotte, P.A.; Bouska,J.J.; Murphy, S.S.;
Davidsen, S.K., Bioorg. Med. Chem. Lett., 2002, 12, 3443.
63.Finnin, M.S.; Donigian, J.R.; Cohen, A.; Richon, V.M.;
Rifkind, R.A.; Marks, P.A.; Pavletich, N. P., Nature,
1999, 401, 188.
64.Somoza, J.R.; Skene, R.J.; Katz, B.A.; Mol, C.; Ho,
J.D., Structure, 2004, 12, 1325.
65.Vannini, A.; Volpari, C.; Filocamo, G.; Casavola, E. C.;
Brunetti, M.; Renzoni, D.; Chakravarty, P.; Paolini, C.;
De Francesco, R.; Gallinari, P.; Steinkuhler, C.; Di
Marco, S., Proc. Natl. Acad. Sci., 2004, 101, 15064.
66.Vannini, A.; Volpari, C.; Filocamo, G.; Casavola, E.C.;
Brunetti, M.; Renzoni, D.; Chakravarty, P.; Paolini, C.;
De Francesco, R.; Gallinari, P.; Steinkuhler, C.; Di
Marco, S., Proc. Natl. Acad. Sci. USA 2004, 101, 15064.
67.Schuetz, A.; Min, J.; Allali-Hassani, A.; Schapira, M.;
Shuen, M.; Loppnau, P.; Mazitschek, R.; Kwiatkowski,
N.P.; Lewis, T.A.; Maglathin, R.L.; McLean, T. H.;
Bochkarev, A.; Plotnikov, A.N.; Vedadi, M.; Arrowsmith,
C.H., J Biol Chem, 2008, 283, 11355.
68.You, L.; Feng, S.; An, R.; Wang, X., Nat. Prod. Commun.,
2009, 4, 297.
69.Huang, R.L.; Chen, C.C.; Huang, Y.L.; Hsieh, D.J.; Hu,
C.P.; Chen, C.F.; Chang, C., Hepatology, 1996, 24, 508.
70.Zhang, Q.; Qin, L.; He, W.; Van Puyvelde, L.; Maes, D.;
Adams, A.; Zheng, H.; De Kimpe, N., Planta Med., 2007,
73, 13.
71.Fujioka, T.; Furumi, K.; Fujii, H.; Okabe, H.; Mihashi,
K.; Nakano, Y.; Matsunaga, H.; Katano, M.; Mori, M.,
Chem. Pharm. Bull., 1999, 47, 96.
72.Kuo, P.L.; Hsu, Y.L.; Chang, C.H.; Chang, J.K., J.
Pharmacol. Exp. Ther., 2005. 314, 1290.
73.Tosun, A.; Akkol, E.K.; Yesilada, E., Z. Naturforsch.
C., 2009, 64, 56. |
| Appears in Collections: | [Graduate Institute of Pharmacognosy Science] Dissertation/Thesis
|
Files in This Item:
There are no files associated with this item.
|
All items in TMUIR are protected by copyright, with all rights reserved.
|
