曹颖莉 李少雄 陈 虹 郭 颖. 非核苷类逆转录酶抑制剂耐药型HIV-1药理评价体系的建立J. 药学学报, 2009,44(4): 355-361.
引用本文: 曹颖莉 李少雄 陈 虹 郭 颖. 非核苷类逆转录酶抑制剂耐药型HIV-1药理评价体系的建立J. 药学学报, 2009,44(4): 355-361.
CAO Ying-Chi, Li-Shao-Xiong, Chen- Gong, Guo- Ying. Establishment of pharmacological evaluation system for non-nucleoside reverse-transcriptase inhibitors resistant HIV-1J. 药学学报, 2009,44(4): 355-361.
Citation: CAO Ying-Chi, Li-Shao-Xiong, Chen- Gong, Guo- Ying. Establishment of pharmacological evaluation system for non-nucleoside reverse-transcriptase inhibitors resistant HIV-1J. 药学学报, 2009,44(4): 355-361.

非核苷类逆转录酶抑制剂耐药型HIV-1药理评价体系的建立

Establishment of pharmacological evaluation system for non-nucleoside reverse-transcriptase inhibitors resistant HIV-1

  • 摘要:

    建立9种临床常见的对非核苷类逆转录酶抑制剂 (non-nucleoside reverse-transcriptase inhibitors, NNRTIs) 耐药型HIV-1重组病毒药理评价模型。应用重叠PCR定点突变技术将耐药突变位点引入HIV-1核心基因 (pNL4-3.Luc.R-E-), 以水泡性口膜炎病毒的外壳糖蛋白 (vesicular stomatitis virus glycoprotein, VSV-G) 包装含耐药突变位点的HIV-1核心,形成耐药型HIV-1重组假病毒颗粒,简称VSVG/HIV-mut (包括VSVG/HIV-wtVSVG/HIV-K103NVSVG/HIV-Y181C
    VSVG/HIV-L100I,K103NVSVG/HIV-Y188LVSVG/HIV-K103N,Y181CVSVG/HIV-K103N,P225HVSVG/HIV-K103N,Y188LVSVG/HIV-K103N,G109AVSVG/HIV-K103N,V108I)。经验证9NNRTIs耐药型HIV-1重组病毒颗粒均具有高感染能力,对核苷类阳性药物不耐药,而对非核苷类阳性药物呈现不同程度的耐药性 (1710 000),且耐药倍数与报道的数据基本一致。所建立的耐药型模型是针对NNRTIs耐药的HIV-1复制环节的细胞水平药效学评价体系,野生型和9种耐药型HIV-1模型的联合应用,可为新型非核苷类逆转录酶抑制剂的研发提供更全面药效学评价的安全平台。

     

    Abstract:

    Consistent non-nucleoside reverse-transcriptase inhibitors (NNRTIs) resistant HIV-1 strains    occurred due to the clinical use for more than ten years of efavirenz (EFV), nevirapine (NVP), and delavirdine (DLV).  In this study, we established nine cell-based pharmacological models according to most NNRTIs-resistant clinical tested strains.  Resistant mutations were introduced into vector, pNL4-3.Luc.R-E-, by overlapping PCR.  Then, pseudovirions were produced by co-transfection of VSV-G plasmid and pNL4-3.Luc.R-E--mut.  All nine recombinant VSVG/HIV-mut pseudovirions (VSVG/HIV-wt, VSVG/HIV-K103N, VSVG/HIV-Y181C, VSVG/ HIV-L100I,K103N, VSVG/HIV-Y188L, VSVG/HIV-K103N,Y181C, VSVG/HIV-K103N,P225H, VSVG/HIV-K103N,Y188L, VSVG/ HIV-K103N,G109A and  
    VSVG/HIV-K103N,V108I) had high efficient infectivity.  Furthermore, they all showed resistant characteristics to EFV and NVP with IC50 changes consisting with clinical reports, not to nucleoside  reverse-transcriptase inhibitors (AZT and d4T).  This series safe cell-based model, which could be carried out in BSL-2 laboratory, can be used for evaluating NNRTIs candidates.

     

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