胆固醇修饰的非天然HR序列肽类HIV-1融合抑制剂的合成及活性初筛

doi:10.7503/cjcu20140572

高等学校化学学报 ›› 2014, Vol. 35 ›› Issue (12): 2542.doi: 10.7503/cjcu20140572

胆固醇修饰的非天然HR序列肽类HIV-1融合抑制剂的合成及活性初筛

张沙, 史卫国(), 王潮, 蔡利锋, 郑保华, 王昆, 冯思良, 贾启燕, 刘克良()

军事医学科学院毒物药物研究所, 北京100850

收稿日期:2014-06-23 出版日期:2014-12-10 发布日期:2014-11-29
作者简介:联系人简介: 刘克良, 男, 博士, 研究员, 博士生导师, 主要从事多肽药物化学、药用高分子材料及核酸化学研究. E-mail:keliangliu@yahoo.com;史卫国, 男, 博士, 副研究员, 主要从事多肽药物化学、新药设计与合成研究. E-mail:shiwg1988@126.com
基金资助:
国家自然科学基金(批准号: 81373266)和“重大新药创制”科技重大专项项目(批准号: 2012ZX09301003)资助

Synthesis of Cholesterol Conjugated Non-native Derived Heptad Repeat Sequence Peptides as Potent HIV-1 Fusion Inhibitors^†

ZHANG Sha, SHI Weiguo^*(), WANG Chao, CAI Lifeng, ZHENG Baohua, WANG Kun, FENG Siliang, JIA Qiyan, LIU Keliang^*()

Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China

Received:2014-06-23 Online:2014-12-10 Published:2014-11-29
Contact: SHI Weiguo,LIU Keliang E-mail:shiwg1988@126.com;keliangliu@yahoo.com
Supported by:
† Supported by the National Natural Science Foundation of China(No.81373266) and the "Creation of Major New Drugs" Science and Technology Major Projects, China(No.2012ZX09301003)

摘要/Abstract

摘要：

将胆固醇分子通过1个半胱氨酸侧链硫醚键和1个β-丙氨酸连接臂引入到所设计的非天然HR序列抗HIV融合活性多肽的C端和N端, 合成了与天然C肽序列同源性很低的非天然序列的类肽抗HIV融合分子, 以考察胆固醇修饰对非天然HR序列活性的影响, 探讨克服耐药性的新思路. 生物活性评价结果表明, 胆固醇与HR肽C端结合物抑制HIV融合活性显著提高, 而连接到N端的序列则完全失去了抗病毒活性, 表明所设计的非天然序列确实具有与天然序列类似的作用机制.

关键词: HIV融合抑制剂, 多肽, 耐药性, 非天然序列

Abstract:

As addition of a cholesterol group to nature peptide fusion inhibitor can dramatically increase its antiviral potency, we designed and synthesized a series of cholesterol conjugated artificial peptide with low sequence homologous from HIV-1 gp41 as HIV-1 fusion inhibitors. The cholesterol moiety was introduced into C- or N-terminal of the peptide through a thioether in cysteine side chain and a β-alanine linker to study the effect of cholesterol modification to the activity of non-natural HR sequence and explore new method to overcome the problem of HIV drug resistance. Cell-cell fusion assays showed that the fusion inhibitory activity of the C-terminal cholesterol conjugated peptide was significantly increased, while the activity of N-terminal conjugated peptide was completely lost, consistent with the reported peptide conjugation using native peptide sequence, indicated that the designed non-natural sequences had a similar mechanism of action with natural sequences. This work provided a useful base for further drug optimization and discovery of highly active non-natural peptide sequences.

Key words: HIV fusion inhibitor, Peptide, Drug resistance, Non-natural sequence

中图分类号:

O629.72

TrendMD:

张沙, 史卫国, 王潮, 蔡利锋, 郑保华, 王昆, 冯思良, 贾启燕, 刘克良. 胆固醇修饰的非天然HR序列肽类HIV-1融合抑制剂的合成及活性初筛. 高等学校化学学报, 2014, 35(12): 2542.

ZHANG Sha, SHI Weiguo, WANG Chao, CAI Lifeng, ZHENG Baohua, WANG Kun, FENG Siliang, JIA Qiyan, LIU Keliang. Synthesis of Cholesterol Conjugated Non-native Derived Heptad Repeat Sequence Peptides as Potent HIV-1 Fusion Inhibitors^†. Chem. J. Chinese Universities, 2014, 35(12): 2542.

图/表 2

参考文献 21

[1]	Chan D. C., Fass D., Berger J. M., Kim P. S., Cell,1997, 89(2), 263—273
[2]	Lu M., Blacklow S. C., Kim P. S., Nat. Struct. Biol., 1995, 2(12), 1075—1082
[3]	Melikyan G. B., Markosyan R. M., Hemmati H., Delmedico M. K., Lambert D. M., Cohen F. S., J. Cell Biol., 2000, 151(2), 413—423
[4]	Kliger Y., Gallo S. A., Peisajovich S. G., Munoz-Barroso I., Avkin S., Blumenthal R., Shai Y., J. Biol. Chem., 2001, 276(2), 1391—1397
[5]	Kaushik-Basu N., Basu A., Harris D., BioDrugs,2008, 22(3), 161—175
[6]	Wei X., Decker J. M., Liu H., Zhang Z., Arani R. B., Kilby J. M., Saag M. S., Wu X., Shaw G. M., Kappes J. C., Antimicrob. Agents Chemother., 2002, 46, 1896—1905
[7]	Greenberg M. L., Cammack N., J. Antimicrob. Chemother., 2004, 54, 333—340
[8]	Xu L., Pozniak A., Wildfire A., Stanfield-Oakley S. A., Mosier S. M., Ratcliffe D., Workman J., Joall A., Myers R., Smit E., Cane P. A., Greenberg M. L., Pillay D., Antimicrob. Agents Chemother., 2005, 49, 1113—1119
[9]	Perez-Alvarez L., Carmona R., Ocampo A., Asorey A., Miralles C., Perez C., Pinilla M., Contreras G., Taboada J. A., Najera R., J. Med. Virol., 2006, 78, 141—147
[10]	Ling Y. B., Wang K., Jiang X. F., Cai L. F., Liu K. L., Chem. J. Chinese Universities, 2013, 34(9), 2102—2107
	(凌彦博, 王昆, 姜喜凤, 蔡利锋, 刘克良. 高等学校化学学报, 2013, 34(9), 2102—2107)
[11]	Xin J. L., Zhang Y., Zhang L. Z., Lin Y., Zhou Q. L., Chem. Res. Chinese Universities, 2013, 29(5), 924—928
[12]	Wang Y. X., Wang Y., Han W. W., Feng Y., Chem. Res. Chinese Universities, 2012, 28(4), 707—711
[13]	Qi Z., Shi W., Xue N., Pan C., Jing W., Liu K., Jiang S., J. Biol. Chem., 2008, 283(44), 30376—30384
[14]	Shi W., Qi Z., Pan C., Xue N., Debnath A. K, Qie J., Jiang S., Liu K., Biochem. Bioph. Res. Co., 2008, 374(4), 767—772
[15]	Wang C., Shi W., Cai L., Lu L., Yu F., Wang Q., Jiang X., Xu X., Wang K., Xu L., Jiang S., Liu K., J. Antimicrob. Chemother., 2014, 69, 1537—1545
[16]	Wang C., Shi W., Cai L., Lu L., Wang Q., Zhang T., Li J., Zhang Z., Wang K., Xu L., Jiang X., Jiang S., Liu K., J. Med. Chem., 2013, 56(6), 2527—2539
[17]	Ingallinella P., Bianchi E., Ladwa N. A., Wang Y., Hrin R., Venezianoc M., Bonellic F., Ketasd T. J., Moored J. P., Millerb M. D., Pessia A., Proc. Natl. Acad. Sci. USA,2009, 106(14), 5801—5806
[18]	Pan C., Cai L., Lu H., Qi Z., Jiang S., J. Virol., 2009, 83(16), 7862—7872
[19]	Liu S., Jing W., Cheung B., Lu H., Sun J., Yan X., Niu J., Farmar J., Wu S., Jiang S., J. Biol. Chem., 2007, 282, 9612—9620
[20]	Otaka A., Nakamura M., Nameki D., Kodama E., Uchiyama S., Nakamura S., Nakano H., Tamamura H., Kobayashi Y., Matsuoka M., Fujii N., Angew. Chem. Int. Ed. Engl., 2002, 41(16), 2937—2940
[21]	Oishi S., Ito S., Nishikawa H., Watanabe K., Tanaka M., Ohno H., Izumi K., Sakagami Y., Kodama E., Matsuoka M., Fujii N., J. Med. Chem., 2008, 51(3), 388—391

Peptide	Sequence	M_w(calcd.)	MALDI-TOF-MS, m/z	Purity by HPLC(%)
4HR	(IEEYTKK)₄	3627	3626.8	>95
5HR	(IEEYTKK)₅	4519	4518.2	>95
4HR-βAla-C	(IEEYTKK)₄-βAla-C	3801	3801.5	>95
5HR-βAla-C	(IEEYTKK)₅-βAla-C	4693	4693.2	>95
C-βAla-4HR	C-βAla-(IEEYTKK)₄	3801	3800.9	>95
C-βAla-5HR	C-βAla-(IEEYTKK)₅	4693	4693.1	>95

Peptide	Sequence	M_w(calcd.)	MALDI-TOF-MS, m/z	Purity by HPLC(%)
4HR	(IEEYTKK)₄	3627	3626.8	>95
5HR	(IEEYTKK)₅	4519	4518.2	>95
4HR-βAla-C	(IEEYTKK)₄-βAla-C	3801	3801.5	>95
5HR-βAla-C	(IEEYTKK)₅-βAla-C	4693	4693.2	>95
C-βAla-4HR	C-βAla-(IEEYTKK)₄	3801	3800.9	>95
C-βAla-5HR	C-βAla-(IEEYTKK)₅	4693	4693.1	>95

Peptide	Sequence	M_w(calcd.)	MALDI-TOF-MS, m/z	Purity by HPLC^a(%)	Inhibiting cell fusion at 200 nmol/L(%)
4HR	(IEEYTKK)₄	3627	3626.8	96.1	ND^b
5HR	(IEEYTKK)₅	4519	4518.2	95.4	10
4HR-βAla-C-Chol	(IEEYTKK)₄-βAla-C-Chol	4228	4227.9	95.3	42
5HR-βAla-C-Chol	(IEEYTKK)₅-βAla-C-Chol	5119	5118.3	94.7	30
Chol-C-βAla-4HR	Chol-C-βAla-(IEEYTKK)₄	4185	4183.8	92.3	0
Chol-C-βAla-5HR	Chol-C-βAla-(IEEYTKK)₅	5077	5077.5	93.1	0
T-20	YTSLIHSLIEESQNQQEKNEQE	4492	4491.9	95.3	98
	LLELDKWASLWNWF

Peptide	Sequence	M_w(calcd.)	MALDI-TOF-MS, m/z	Purity by HPLC^a(%)	Inhibiting cell fusion at 200 nmol/L(%)
4HR	(IEEYTKK)₄	3627	3626.8	96.1	ND^b
5HR	(IEEYTKK)₅	4519	4518.2	95.4	10
4HR-βAla-C-Chol	(IEEYTKK)₄-βAla-C-Chol	4228	4227.9	95.3	42
5HR-βAla-C-Chol	(IEEYTKK)₅-βAla-C-Chol	5119	5118.3	94.7	30
Chol-C-βAla-4HR	Chol-C-βAla-(IEEYTKK)₄	4185	4183.8	92.3	0
Chol-C-βAla-5HR	Chol-C-βAla-(IEEYTKK)₅	5077	5077.5	93.1	0
T-20	YTSLIHSLIEESQNQQEKNEQE	4492	4491.9	95.3	98
	LLELDKWASLWNWF

胆固醇修饰的非天然HR序列肽类HIV-1融合抑制剂的合成及活性初筛

Synthesis of Cholesterol Conjugated Non-native Derived Heptad Repeat Sequence Peptides as Potent HIV-1 Fusion Inhibitors^†

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胆固醇修饰的非天然HR序列肽类HIV-1融合抑制剂的合成及活性初筛

Synthesis of Cholesterol Conjugated Non-native Derived Heptad Repeat Sequence Peptides as Potent HIV-1 Fusion Inhibitors†

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Synthesis of Cholesterol Conjugated Non-native Derived Heptad Repeat Sequence Peptides as Potent HIV-1 Fusion Inhibitors^†