以HIV-1 IN-LEDGF/p75相互作用为靶点的抑制剂筛选
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摘要
目的晶状体上皮源性生长因子p~(75)蛋白(LEDGF/p75)与HIV-1整合酶(IN)之间的蛋白-蛋白相互作用是开发抗HIV-1药物的有效靶点,本文旨在寻找以HIV-1IN-LEDGF/p75相互作用为靶点的小分子抑制剂。方法采用均相时间分辨荧光技术(HTRF)筛选了799个化合物,比对IN-LEDGF/p75相互作用的抑制活性。结果发现5个化合物——肾上腺酮、6-溴-1,2-二氢萘-1,2-二酮、头孢噻吩、根皮含柘树呫吨酮L和迷迭香酸对该相互作用表现出不同程度的抑制作用,其IC50值分别为12.3、22.7、26.2、18.5和1.13μmol/L。结论为HIV-1IN-LEDGF/p75相互作用抑制剂的发现以及新的抗HIV-1药物的开发提供了重要基础。
Objective The protein-protein interaction between the cellular chromatin associated protein lens epithelium-derived growth factor(LEDGF/p75) and the HIV-1 integrase(IN) is an effective target for the development of anti-HIV-1 drugs. This work aims to find small molecular inhibitors targeting the HIV-1 IN-LEDGF/p75 interaction.Methods The inhibitory activity of 799 compounds(from 2 compounds libraries: InterBioScreen and BioBioPha) on the interaction of IN-LEDGF/p75 was screened by a homogeneous time resolved fluorescence(HTRF)-based assay.Results It was found that five compounds, adrenalone, bonaphton, cefalotin, cudraxanthone L and rosmarinic acid, showed different inhibitory effect on the interaction of HIV-1 IN-LEDGF/p75, and their IC50 values were 12.3, 22.7, 26.2, 18.5 and 1.13 μmol/L, respectively. Conclusion This work provides an important basis for the discovery of HIV-1 IN-LEDGF/p75 interaction inhibitors and the development of new anti-HIV-1 drugs.
Objective The protein-protein interaction between the cellular chromatin associated protein lens epithelium-derived growth factor(LEDGF/p75) and the HIV-1 integrase(IN) is an effective target for the development of anti-HIV-1 drugs. This work aims to find small molecular inhibitors targeting the HIV-1 IN-LEDGF/p75 interaction.Methods The inhibitory activity of 799 compounds(from 2 compounds libraries: InterBioScreen and BioBioPha) on the interaction of IN-LEDGF/p75 was screened by a homogeneous time resolved fluorescence(HTRF)-based assay.Results It was found that five compounds, adrenalone, bonaphton, cefalotin, cudraxanthone L and rosmarinic acid, showed different inhibitory effect on the interaction of HIV-1 IN-LEDGF/p75, and their IC50 values were 12.3, 22.7, 26.2, 18.5 and 1.13 μmol/L, respectively. Conclusion This work provides an important basis for the discovery of HIV-1 IN-LEDGF/p75 interaction inhibitors and the development of new anti-HIV-1 drugs.
引文
[1]Gilbert PB,Mckeague IW,Eisen G,et al.Comparison of HIV-1 and HIV-2 infectivity from a prospective cohort study in Senegal.Stat Med,2003,22(4):573-593.
[2]Zhang X.Anti-retroviral drugs:current state and development in the next decade.Acta Pharm Sin B,2018,8(2):131-136.
[3]Dayam R,Al-Mawsawi LQ,Neamati DN.HIV-1 integrase inhibitors:an emerging clinical reality.Drugs R D,2007,8(3):155-168.
[4]Hazuda DJ.HIV integrase as a target for antiretroviral therapy.Curr Opin HIV AIDS,2012,7(5):383-389.
[5]Hazuda DJ,Felock P,Witmer M,et al.Inhibitors of strand transfer that prevent integration and inhibit HIV-1 replication in cells.Science,2000,287(5453):646-650.
[6]Métifiot M,Vandegraaff N,Maddali K,et al.Elvitegravir overcomes resistance to raltegravir induced by integrase mutation Y143.AIDS,2011,25(9):1175-1178.
[7]Van Maele B,Busschots K,Vandekerckhove L,et al.Cellular co-factors of HIV-1 integration.Trends Biochem Sci,2006,31(2):98-105.
[8]Maertens G,Cherepanov P,Pluymers W,et al.LEDGF/p75 is essential for nuclear and chromosomal targeting of HIV-1 integrase in human cells.J Biol Chem,2003,278(35):33528-33539.
[9]De Rijck J,Vandekerckhove L,Gijsbers R,et al.Overexpression of the lens epithelium-derived growth factor/p75 integrase binding domain inhibits human immunodeficiency virus replication.J Virol,2006,80(23):11498-11509.
[10]Wang HJ,Zhou ZH,Xu JC,et al.HTRF-based method for determination of HSP90-HOP inhibition activity and its application.Acta Pharm Sinica,2017,52(4):592-597.(in Chinese)王慧洁,周子涵,徐嘉辰,等.基于均相时间分辨荧光技术(HTRF)的HSP90-HOP相互作用抑制剂活性测试方法的构建及其应用.药学学报,2017,52(4):592-597.
[11]Fotopoulou MA,Ioannou PC.Post-column terbium complexation and sensitized fluorescence detection for the determination of norepinephrine,epinephrine and dopamine using high-performance liquid chromatography.Anal Chimica Acta,2002,462(2):179-185.
[12]Nikolaeva IS,Kutchak SN,Bogdanova NS,et al.Pathomorphological study of antiviral chemotherapeutic effectiveness of bonaphthon under experimental conditions.Farmakol Toksikol,1976,39(3):336-341.
[13]Carranza-Torres JM,Vargas-Daza ER,Villarreal-Ríos E,et al.Effectiveness of the scheme reimpregnation maintenance schedule vs.ceftazidime/cephalothin in dialysis patients with peritonitis.Nefrologia,2016,36(4):448-450.
[14]Lian SS.Research progress on xanthones.Guangdong Chem Ind,2014,41(2):46-47.(in Chinese)连双双.氧杂蒽酮类化合物的研究进展.广东化工,2014,41(2):46-47.
[15]You R,Ma XQ,Wu BH,et al.Advance in pharmacological effects of Rosmarinic Acid.Sichuan J Physiol Sci,2015,37(2):93-96.(in Chinese)尤茹,马雪倩,吴炳火,等.迷迭香酸药理作用研究进展.四川生理科学杂志,2015,37(2):93-96.
[16]Tewtrakul S,Miyashiro H,Nakamura N,et al.HIV-1 integrase inhibitory substances from Coleus parvifolius.Phytother Res,2003,17(3):232-239.
[2]Zhang X.Anti-retroviral drugs:current state and development in the next decade.Acta Pharm Sin B,2018,8(2):131-136.
[3]Dayam R,Al-Mawsawi LQ,Neamati DN.HIV-1 integrase inhibitors:an emerging clinical reality.Drugs R D,2007,8(3):155-168.
[4]Hazuda DJ.HIV integrase as a target for antiretroviral therapy.Curr Opin HIV AIDS,2012,7(5):383-389.
[5]Hazuda DJ,Felock P,Witmer M,et al.Inhibitors of strand transfer that prevent integration and inhibit HIV-1 replication in cells.Science,2000,287(5453):646-650.
[6]Métifiot M,Vandegraaff N,Maddali K,et al.Elvitegravir overcomes resistance to raltegravir induced by integrase mutation Y143.AIDS,2011,25(9):1175-1178.
[7]Van Maele B,Busschots K,Vandekerckhove L,et al.Cellular co-factors of HIV-1 integration.Trends Biochem Sci,2006,31(2):98-105.
[8]Maertens G,Cherepanov P,Pluymers W,et al.LEDGF/p75 is essential for nuclear and chromosomal targeting of HIV-1 integrase in human cells.J Biol Chem,2003,278(35):33528-33539.
[9]De Rijck J,Vandekerckhove L,Gijsbers R,et al.Overexpression of the lens epithelium-derived growth factor/p75 integrase binding domain inhibits human immunodeficiency virus replication.J Virol,2006,80(23):11498-11509.
[10]Wang HJ,Zhou ZH,Xu JC,et al.HTRF-based method for determination of HSP90-HOP inhibition activity and its application.Acta Pharm Sinica,2017,52(4):592-597.(in Chinese)王慧洁,周子涵,徐嘉辰,等.基于均相时间分辨荧光技术(HTRF)的HSP90-HOP相互作用抑制剂活性测试方法的构建及其应用.药学学报,2017,52(4):592-597.
[11]Fotopoulou MA,Ioannou PC.Post-column terbium complexation and sensitized fluorescence detection for the determination of norepinephrine,epinephrine and dopamine using high-performance liquid chromatography.Anal Chimica Acta,2002,462(2):179-185.
[12]Nikolaeva IS,Kutchak SN,Bogdanova NS,et al.Pathomorphological study of antiviral chemotherapeutic effectiveness of bonaphthon under experimental conditions.Farmakol Toksikol,1976,39(3):336-341.
[13]Carranza-Torres JM,Vargas-Daza ER,Villarreal-Ríos E,et al.Effectiveness of the scheme reimpregnation maintenance schedule vs.ceftazidime/cephalothin in dialysis patients with peritonitis.Nefrologia,2016,36(4):448-450.
[14]Lian SS.Research progress on xanthones.Guangdong Chem Ind,2014,41(2):46-47.(in Chinese)连双双.氧杂蒽酮类化合物的研究进展.广东化工,2014,41(2):46-47.
[15]You R,Ma XQ,Wu BH,et al.Advance in pharmacological effects of Rosmarinic Acid.Sichuan J Physiol Sci,2015,37(2):93-96.(in Chinese)尤茹,马雪倩,吴炳火,等.迷迭香酸药理作用研究进展.四川生理科学杂志,2015,37(2):93-96.
[16]Tewtrakul S,Miyashiro H,Nakamura N,et al.HIV-1 integrase inhibitory substances from Coleus parvifolius.Phytother Res,2003,17(3):232-239.