靛红类双功能抑制剂: 调控SARS-3CL蛋白酶聚集状态与活性

引用本文: 周璐, 金凤, 刘莹, 尚尔昌, 魏平, 李春梅, 来鲁华. 靛红类双功能抑制剂: 调控SARS-3CL蛋白酶聚集状态与活性[J]. 物理化学学报, 2012, 28(10): 2418-2422. doi: 10.3866/PKU.WHXB201209143 shu

Citation: ZHOU Lu, JIN Feng, LIU Ying, SHANG Er-Chang, WEI Ping, LI Chun-Mei, LAI Lu-Hua. Isatin Dual Functional Inhibitors: Modulating the Aggregation State and Enzyme Activity of SARS-3CL Proteinase[J]. Acta Physico-Chimica Sinica, 2012, 28(10): 2418-2422. doi: 10.3866/PKU.WHXB201209143 shu

靛红类双功能抑制剂: 调控SARS-3CL蛋白酶聚集状态与活性

周璐 ^1,3, ,
金凤 ^1, ,
刘莹 ^1,2, ,
尚尔昌 ^1, ,
魏平 ^1, ,
李春梅 ^1, ,
来鲁华 ^1,2,

基金项目:
国家自然科学基金(90913021, 20473001, 11021463) (90913021, 20473001, 11021463)

国家重点基础研究发展规划项目(973) (2009CB9185003)资助 (973) (2009CB9185003)

摘要:

1-(2-萘甲基)靛红-5-甲酰胺类化合物通过与底物口袋结合来抑制SARS-3CL 蛋白酶的活性, 而SARS-3CL蛋白酶自身的N端8 肽是作用于蛋白二聚界面的抑制剂. 本文设计同时占据SARS-3CL蛋白酶底物口袋和二聚界面的双功能抑制剂, 通过固相多肽合成方法制备由1-(2-萘甲基)靛红-5-甲酸和N端8肽组成的化合物, 得到不同长度连接链的6 个目标产物. 用显色底物方法测定化合物对SARS-3CL蛋白酶的抑制活性,其中化合物3的活性最高, IC₅₀值(半抑制率)为3.8 μmol·L^-1, 连接偶数甘氨酸的活性明显要好于连接奇数甘氨酸的化合物. 用超速离心沉降速率方法研究了化合物3对SARS-3CL蛋白酶聚集状态与活性的调控作用, 其同时具有诱导与抑制二聚的双重能力, 综合调控结果是抑制SARS-3CL蛋白酶的二聚. 这项研究给应用合成的化合物研究酶活性调节机制提供了一个示例.

关键词:

English

Isatin Dual Functional Inhibitors: Modulating the Aggregation State and Enzyme Activity of SARS-3CL Proteinase

1.
1 Beijing National Laboratory for Molecular Sciences, State Key Laboratory of Structural Chemistry of Unstable and Stable Species, Institute of Physical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China;
2 Center for Quantitative Biology, Peking University, Beijing 100871, P. R. China;
3 School of Pharmacy, Fudan University, Shanghai 201203, P. R. China
Received Date: 25 July 2012
Available Online: 26 September 2012
Fund Project:
国家自然科学基金(90913021, 20473001, 11021463)

国家重点基础研究发展规划项目(973) (2009CB9185003)资助

Abstract:

The 1-(2-naphthlmethyl) isatin-5-formamide compounds can inhibit SARS-3CL proteinase by binding to its substrate pocket, while the N-terminal octapeptide of SARS-3CL proteinase was found to act as a dimerization inhibitor. In this work, the dual functional inhibitors which can occupy both substrate pocket of SARS-3CL proteinase and its dimer interface were designed. Six title compounds were tten by linking 1-(2-naphthlmethyl) isatin-5-formic acid and N-terminal octapeptides using a polyglycine linker through solid-phase peptide synthesis method. The in vitro inhibition activity against SARS-3CL proteinase was measured by continuous colorimetric assay using colorimetric substrate. Compound 3 showed the highest inhibition activity with an IC₅₀ (half maximal inhibitory concentration of a substance) of 3.8 μmol·L^-1. The change of inhibition activity with the linker length was studied. Inhibitors with the even spacers were showed better activity than the odd ones, which could be explained by the angle restriction of peptide bonds. The modulating of the aggregation state and enzyme activity towards SARS-3CL proteinase were studied using sedimentation velocity experiments. Compound 3 was found to not only inhibit the enzyme activity of SARS-3CL proteinase, but also shift the monomer-dimer equilibrium of the enzyme. The integrated control result is inhibiting SARS-3CL proteinase dimer formation. This work provides an example of using synthesized compounds to study enzyme activity regulation mechanism.

Key words:

1. [1]
  
  (1) Bogert, M. T.; Nabenhauer, F. P. J. Am. Chem. Soc. 1924, 46,1702. doi: 10.1021/ja01672a021
  (1) Bogert, M. T.; Nabenhauer, F. P. J. Am. Chem. Soc. 1924, 46,1702. doi: 10.1021/ja01672a021
2. [2]
  (2) Trinka, P.; Slegel, P.; Reiter, J. J. Prakt. Chem. 1996, 338, 675.doi: 10.1002/(ISSN)1521-3897(2) Trinka, P.; Slegel, P.; Reiter, J. J. Prakt. Chem. 1996, 338, 675.doi: 10.1002/(ISSN)1521-3897
3. [3]
  (3) Praveen, C.; Ayyanar, A.; Perumal, P. T. Bioorg. Med. Chem. Lett. 2011, 21, 4072. doi: 10.1016/j.bmcl.2011.04.117(3) Praveen, C.; Ayyanar, A.; Perumal, P. T. Bioorg. Med. Chem. Lett. 2011, 21, 4072. doi: 10.1016/j.bmcl.2011.04.117
4. [4]
  (4) Shingade, S. G.; Bari, S. B.;Waghmare, U. B. Med. Chem. Res.2012, 21, 1302. doi: 10.1007/s00044-011-9644-y(4) Shingade, S. G.; Bari, S. B.;Waghmare, U. B. Med. Chem. Res.2012, 21, 1302. doi: 10.1007/s00044-011-9644-y
5. [5]
  (5) Adibi, H.; Khodaei, M.; Pakravan, P.; Abiri, R. Pharm. Chem. J.2010, 44, 219. doi: 10.1007/s11094-010-0436-3(5) Adibi, H.; Khodaei, M.; Pakravan, P.; Abiri, R. Pharm. Chem. J.2010, 44, 219. doi: 10.1007/s11094-010-0436-3
6. [6]
  (6) Pervez, H.; Iqbal, M. S.; Tahir, M. Y.; Nasim, F. U.; Choudhary,M. I.; Khan, K. M. J. Enzym. Inhib. Med. Chem. 2008, 23, 848.doi: 10.1080/14756360701746179(6) Pervez, H.; Iqbal, M. S.; Tahir, M. Y.; Nasim, F. U.; Choudhary,M. I.; Khan, K. M. J. Enzym. Inhib. Med. Chem. 2008, 23, 848.doi: 10.1080/14756360701746179
7. [7]
  (7) Pervez, H.; Lqbal, M. S.; Tahir, M. Y.; Choudhary, M. I.; Khan,K. M. Nat. Prod. Res. 2007, 21, 1178. doi: 10.1080/14786410601129770(7) Pervez, H.; Lqbal, M. S.; Tahir, M. Y.; Choudhary, M. I.; Khan,K. M. Nat. Prod. Res. 2007, 21, 1178. doi: 10.1080/14786410601129770
8. [8]
  (8) Dra vich, P. S.; Prins, T. J.; Zhou, R.;Webber, S. E.;Marakovits, J. T.; Fuhrman, S. A.; Patick, A. K.; Matthews, D.A.; Lee, C. A.; Ford, C. E.; Burke, B. J.; Rejto, P. A.;Hendrickson, T. F.; Brown, E. L.; Meador, J.W.; Ferre, R. A.;Harr, J. E. V.; Kosa, M. B.;Worland, S. T. J. Med. Chem. 1999,42, 1213. doi: 10.1021/jm9805384(8) Dra vich, P. S.; Prins, T. J.; Zhou, R.;Webber, S. E.;Marakovits, J. T.; Fuhrman, S. A.; Patick, A. K.; Matthews, D.A.; Lee, C. A.; Ford, C. E.; Burke, B. J.; Rejto, P. A.;Hendrickson, T. F.; Brown, E. L.; Meador, J.W.; Ferre, R. A.;Harr, J. E. V.; Kosa, M. B.;Worland, S. T. J. Med. Chem. 1999,42, 1213. doi: 10.1021/jm9805384
9. [9]
  (9) Webber, S. E.; Tikhe, J.;Worland, S. T.; Fuhrman, S. A.;Hendrickson, T. F.; Matthews, D. A.; Love, R. A.; Patick, A. K.;Meador, J.W.; Ferre, R. A.; Brown, E. L.; DeLisle, D. M.; Ford,C. E.; Binford, S. L. J. Med. Chem. 1996, 39, 5072. doi: 10.1021/jm960603e(9) Webber, S. E.; Tikhe, J.;Worland, S. T.; Fuhrman, S. A.;Hendrickson, T. F.; Matthews, D. A.; Love, R. A.; Patick, A. K.;Meador, J.W.; Ferre, R. A.; Brown, E. L.; DeLisle, D. M.; Ford,C. E.; Binford, S. L. J. Med. Chem. 1996, 39, 5072. doi: 10.1021/jm960603e
10. [10]
  (10) Vine, K. L.; Matesic, L.; Locke, J. M.; Ranson, M.; Skropeta, D.Anti-Cancer Agents Med. Chem. 2009, 9, 397.(10) Vine, K. L.; Matesic, L.; Locke, J. M.; Ranson, M.; Skropeta, D.Anti-Cancer Agents Med. Chem. 2009, 9, 397.
11. [11]
  (11) Kandile, N. G.; Mohamed, M. I.; Ismaeel, H. M. J. Enzym. Inhib. Med. Chem. 2012, 27, 330. doi: 10.3109/14756366.2011.588950(11) Kandile, N. G.; Mohamed, M. I.; Ismaeel, H. M. J. Enzym. Inhib. Med. Chem. 2012, 27, 330. doi: 10.3109/14756366.2011.588950
12. [12]
  (12) Peiris, J. S. M.; Lai, S. T.; Poon, L. L. M.; Guan, Y.; Yam, L. Y.C.; Lim,W.; Nicholls, J.; Yee,W. K. S.; Yan,W.W.; Cheung,M. T. ; Cheng, V. C. C.; Chan, K. H.; Tsang, D. N. C.; Yung, R.W. H.; Ng, T. K.; Yuen, K. Y. Lancet 2003, 361, 1319. doi: 10.1016/S0140-6736(03)13077-2(12) Peiris, J. S. M.; Lai, S. T.; Poon, L. L. M.; Guan, Y.; Yam, L. Y.C.; Lim,W.; Nicholls, J.; Yee,W. K. S.; Yan,W.W.; Cheung,M. T. ; Cheng, V. C. C.; Chan, K. H.; Tsang, D. N. C.; Yung, R.W. H.; Ng, T. K.; Yuen, K. Y. Lancet 2003, 361, 1319. doi: 10.1016/S0140-6736(03)13077-2
13. [13]
  (13) Yang, H. T.; Yang, M. J.; Ding, Y.; Liu, Y.W.; Lou, Z. Y.; Zhou,Z.; Sun, L.; Mo, L. J.; Ye, S.; Pang, H.; Gao, G. F.; Anand, K.;Bartlam, M.; Hilgenfeld, R.; Rao, Z. H. Proc. Natl. Acad. Sci. U. S. A. 2003, 100, 13190. doi: 10.1073/pnas.1835675100(13) Yang, H. T.; Yang, M. J.; Ding, Y.; Liu, Y.W.; Lou, Z. Y.; Zhou,Z.; Sun, L.; Mo, L. J.; Ye, S.; Pang, H.; Gao, G. F.; Anand, K.;Bartlam, M.; Hilgenfeld, R.; Rao, Z. H. Proc. Natl. Acad. Sci. U. S. A. 2003, 100, 13190. doi: 10.1073/pnas.1835675100
14. [14]
  (14) Lai, L. H.; Han, X. F.; Chen, H.;Wei, P.; Huang, C. K.; Liu, S.Y.; Fan, K. Q.; Zhou, L.; Liu, Z. M.; Pei, J. F.; Liu, Y. Curr. Pharm. Des. 2006, 12, 4555. doi: 10.2174/138161206779010396(14) Lai, L. H.; Han, X. F.; Chen, H.;Wei, P.; Huang, C. K.; Liu, S.Y.; Fan, K. Q.; Zhou, L.; Liu, Z. M.; Pei, J. F.; Liu, Y. Curr. Pharm. Des. 2006, 12, 4555. doi: 10.2174/138161206779010396
15. [15]
  (15) Chen, L. R.;Wang, Y. C.; Lin, Y.W.; Chou, S. Y.; Chen, S. F.;Liu, L. T.;Wu, Y. T.; Chih-Jung, K. B.; Chen, T. S. S.; Juang, S.H. Bioorg. Med. Chem. Lett. 2005, 15, 3058. doi: 10.1016/j.bmcl.2005.04.027(15) Chen, L. R.;Wang, Y. C.; Lin, Y.W.; Chou, S. Y.; Chen, S. F.;Liu, L. T.;Wu, Y. T.; Chih-Jung, K. B.; Chen, T. S. S.; Juang, S.H. Bioorg. Med. Chem. Lett. 2005, 15, 3058. doi: 10.1016/j.bmcl.2005.04.027
16. [16]
  (16) Zhou, L.; Liu, Y.; Zhang,W. L.;Wei, P.; Huang, C. K.; Pei, J. F.;Yuan, Y. X.; Lai, L. H. J. Med. Chem. 2006, 49, 3440. doi: 10.1021/jm0602357(16) Zhou, L.; Liu, Y.; Zhang,W. L.;Wei, P.; Huang, C. K.; Pei, J. F.;Yuan, Y. X.; Lai, L. H. J. Med. Chem. 2006, 49, 3440. doi: 10.1021/jm0602357
17. [17]
  (17) Liu, Y.; Zheng, T. F.; Jin, F.; Zhou, L.; Liu, Z. M.;Wei, P.; Lai,L. H. Acta Chim. Sin. 2007, 65, 1707. [刘莹, 郑腾飞,金凤, 周璐, 刘振明, 魏平, 来鲁华. 化学学报, 2007, 65,1707.](17) Liu, Y.; Zheng, T. F.; Jin, F.; Zhou, L.; Liu, Z. M.;Wei, P.; Lai,L. H. Acta Chim. Sin. 2007, 65, 1707. [刘莹, 郑腾飞,金凤, 周璐, 刘振明, 魏平, 来鲁华. 化学学报, 2007, 65,1707.]
18. [18]
  (18) Fan, K. Q.;Wei, P.; Feng, Q.; Chen, S. D.; Huang, C. K.; Ma,L.; Lai, B.; Pei, J. F.; Liu, Y.; Chen, J. G.; Lai, L. H. J. Biol. Chem. 2004, 279, 1637. doi: 10.1074/jbc.M310875200(18) Fan, K. Q.;Wei, P.; Feng, Q.; Chen, S. D.; Huang, C. K.; Ma,L.; Lai, B.; Pei, J. F.; Liu, Y.; Chen, J. G.; Lai, L. H. J. Biol. Chem. 2004, 279, 1637. doi: 10.1074/jbc.M310875200
19. [19]
  (19) Chen, H.;Wei, P.; Huang, C. K; Tan, L.; Liu, Y.; Lai, L. H.J. Biol. Chem. 2006, 281, 13894. doi: 10.1074/jbc.M510745200(19) Chen, H.;Wei, P.; Huang, C. K; Tan, L.; Liu, Y.; Lai, L. H.J. Biol. Chem. 2006, 281, 13894. doi: 10.1074/jbc.M510745200
20. [20]
  (20) Wei, P.; Fan, K. Q.; Chen, H.; Ma, L.; Huang, C. K.; Tan, L.; Xi,D.; Li, C. M.; Liu, Y.; Cao, A. N.; Lai, L. H. Biochem. Biophys. Res. Commun. 2006, 339, 865. doi: 10.1016/j.bbrc.2005.11.102(20) Wei, P.; Fan, K. Q.; Chen, H.; Ma, L.; Huang, C. K.; Tan, L.; Xi,D.; Li, C. M.; Liu, Y.; Cao, A. N.; Lai, L. H. Biochem. Biophys. Res. Commun. 2006, 339, 865. doi: 10.1016/j.bbrc.2005.11.102
21. [21]
  (21) Wei, P.; Li, C. M.; Zhou, L.; Liu, Y.; Lai, L. H. Acta Phys. -Chim. Sin. 2010, 26, 1093. [魏平, 李春梅, 周璐, 刘莹, 来鲁华. 物理化学学报, 2010, 26, 1093.] doi: 10.3866/PKU.WHXB20100449(21) Wei, P.; Li, C. M.; Zhou, L.; Liu, Y.; Lai, L. H. Acta Phys. -Chim. Sin. 2010, 26, 1093. [魏平, 李春梅, 周璐, 刘莹, 来鲁华. 物理化学学报, 2010, 26, 1093.] doi: 10.3866/PKU.WHXB20100449
22. [22]
  (22) Li, C. M.; Qi, Y. F.; Teng, X.; Yang, Z. C.;Wei, P.; Zhang, C. S.;Tan, L.; Zhou, L.; Liu, Y.; Lai, L. H. J. Biol. Chem. 2010, 285,28134. doi: 10.1074/jbc.M109.095851(22) Li, C. M.; Qi, Y. F.; Teng, X.; Yang, Z. C.;Wei, P.; Zhang, C. S.;Tan, L.; Zhou, L.; Liu, Y.; Lai, L. H. J. Biol. Chem. 2010, 285,28134. doi: 10.1074/jbc.M109.095851
23. [23]
  (23) Fan, K. Q.; Ma, L.; Han, X. F.; Liang, H. H.;Wei, P.; Liu, Y.;Lai, L. H. Biochem. Biophys. Res. Commun. 2005, 329, 934.doi: 10.1016/j.bbrc.2005.02.061(23) Fan, K. Q.; Ma, L.; Han, X. F.; Liang, H. H.;Wei, P.; Liu, Y.;Lai, L. H. Biochem. Biophys. Res. Commun. 2005, 329, 934.doi: 10.1016/j.bbrc.2005.02.061
24. [24]
  (24) Yang, H. T.; Xie,W. Q.; Xue, X. Y.; Yang, K. L.; Ma, J.; Liang,W. X.; Zhao, Q.; Zhou, Z.; Pei, D. Q.; Ziebuhr, J.; Hilgenfeld,R.; Yuen, K. Y.;Wong, L.; Gao, G. X.; Chen, S. J.; Chen, Z.;Ma, D.W.; Bartlam, M.; Rao, Z. H. PLoS Biol. 2005, 3, 1742.(24) Yang, H. T.; Xie,W. Q.; Xue, X. Y.; Yang, K. L.; Ma, J.; Liang,W. X.; Zhao, Q.; Zhou, Z.; Pei, D. Q.; Ziebuhr, J.; Hilgenfeld,R.; Yuen, K. Y.;Wong, L.; Gao, G. X.; Chen, S. J.; Chen, Z.;Ma, D.W.; Bartlam, M.; Rao, Z. H. PLoS Biol. 2005, 3, 1742.
25. [25]
  (25) Huang, C. K.;Wei, P.; Fan, K. Q.; Liu, Y.; Lai, L. H.Biochemistry 2004, 43, 4568. doi: 10.1021/bi036022q
  (25) Huang, C. K.;Wei, P.; Fan, K. Q.; Liu, Y.; Lai, L. H.Biochemistry 2004, 43, 4568. doi: 10.1021/bi036022q

扫一扫看文章

计量

PDF下载量: 506
文章访问数: 2493
HTML全文浏览量: 61

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

靛红类双功能抑制剂: 调控SARS-3CL蛋白酶聚集状态与活性

English

Isatin Dual Functional Inhibitors: Modulating the Aggregation State and Enzyme Activity of SARS-3CL Proteinase

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

靛红类双功能抑制剂: 调控SARS-3CL蛋白酶聚集状态与活性

English

Isatin Dual Functional Inhibitors: Modulating the Aggregation State and Enzyme Activity of SARS-3CL Proteinase

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs