Screening of HIV-1 replication inhibitors by using pseudotyped virus system
- Author:
- No author available
- Journal Title:
- ACTA PHARMACEUTICA SINICA
- Issue:
- 3
- DOI:
- 10.3321/j.issn:0513-4870.2008.03.005
- Key Word:
- HIV-1;药物筛选;假病毒;复制抑制剂
Abstract： 建立以HIV-1复制为靶点的细胞水平假病毒药理筛选模型,并运用该模型筛选化合物库中随机选取的化合物.细胞水平的HIV-1假病毒药理筛选模型是应用水泡性口膜炎病毒的外壳糖蛋白(vesicular stomatitis virus glycoprotein,VSV-G)包装HIV-1核心建立的,简称VSVG/HIV模型.细胞被VSVG/HIV感染后,病毒携带的报告基因的表达水平(荧光素酶活性或GFP阳性细胞比例)反映了HIV-1的复制水平.对HIV-1复制有抑制作用的化合物应用VSVG/MLV模型对其特异性地进一步检测.被感染细胞中报告基因的表达水平与病毒稀释度呈显著的剂量依赖效应.阳性药物齐多夫定(zidovudine,AZT)、拉米夫定(lamivudine,3TC)、去羟基苷(didanosine,DDI)可剂量依赖性地抑制HIV-1的复制,其IC50分别为48.5 nmol·L-1、 0.13 μmol·L-1和1.73 μmol·L-1,与文献报道一致.在随机筛选的500个化合物中有3个化合物可以抑制HIV-1的复制,IC50分别为1.92、 5.38和3.39 μmol·L-1,而对MLV的复制无影响.VSVG/HIV假病毒系统是一种安全、有效的针对HIV-1复制的药理筛选模型.当将此模型与VSVG/MLV模型联合使用时,可判断化合物作用的特异性.实验表明:3个化合物,2-甲硫基-5-(4-甲基苯并)酰胺基-1,3,4-噻二唑(化合物A)、N-(3-羟基苯基)-2-(4-异丁基苯基)丙酰胺(化合物B)和N-(4-甲基吡啶基)-4-甲基苯磺酰胺(化合物C)可以特异性抑制HIV-1的复制.
- [1]杨勤刚,何煦昌,白东鲁.HIV蛋白酶抑制剂研究进展[J].2005,(5).
- [2]Jones PS.Strategies for antiviral drug discovery.[J].1998,9(4).
- [3]Reeves JD,Piefer AJ.Emerging drug targets for antiretroviral therapy.[J].2005,65(13).
- [4]Pauwels R.Aspects of successful drug discovery and development.[J].2006,71(2).
- [5]M, Onishi,S, Kinoshita,Y, Morikawa,etc.Applications of retrovirus-mediated expression cloning.[J].1996,24.
- [6]Landau NR.,Walker R.,Choe S.,etc.Human Immunodeficiency Virus Type 1 Viral Protein R (Vpr) Arrests Cells in the G(2) Phase of the Cell Cycle by Inhibiting P34(Cdc2) Activity[J].1995,69(11).
- [7]Adelson ME,Peltz SW,Pacchia AL,etc.Toward the development of a virus-cell-based assay for the discovery of novel compounds against human immunodeficiency virus type 1.[J].2003,47(2).
- [8]Blair WS,Cao J,Isaacson J,etc.A novel HIV-1 antiviral high throughput screening approach for the discovery of HIV-1 inhibitors.[J].2005,65(2).
- [9]Richman DD,Jacobsen DM,Cooper DA,etc.Antiretroviral Treatment for Adult HIV Infection in 2002: Updated Recommendations of the International AIDS Society-USA Panel.[J].2002,288(2).
- [10]Benjamin Kuan Chen,Ruth I. Connor,Sunny Choe,etc.Vpr Is Required for Efficient Replication of Human Immunodeficiency Virus Type-1 in Mononuclear Phagocytes[J].1995,206(2).
- [11]KATHLEEN A. PAGE,TERI LIEGLER,MARK B. FEINBERG.Use of a Green Fluorescent Protein as a Marker for Human Immunodeficiency Virus Type 1 Infection[J].1997,13(13).
- [12]Blair WS,Cao J,Isaacson J,etc.High-throughput human immunodeficiency virus type 1 (HIV-1) full replication assay that includes HIV-1 Vif as an antiviral target.[J].2005,49(9).
- [13]Griffiths JC,Cannon PM,Ramsdale EE,etc.A transient three-plasmid expression system for the production of high titer retroviral vectors.[J].1995,23(4).
- [14]Pak CC,Puri A,Blumenthal R.Conformational changes and fusion activity of vesicular stomatitis virus glycoprotein: [125I]iodonaphthyl azide photolabeling studies in biological membranes[J].1997,36(29).
- [15]Parkin NT,Winslow GA,Limoli KL,etc.A novel phenotypic drug susceptibility assay for human immunodeficiency virus type 1.[J].2000,44(4).
- [16]Fran?ois, Clavel,Allan J, Hance.HIV drug resistance.[J].2004,350(10).