In Vitro Resistance Selection with Doravirine (MK-1439), a Novel Nonnucleoside Reverse Transcriptase Inhibitor with Distinct Mutation Development Pathways
In Vitro Resistance Selection with Doravirine (MK-1439), a Novel Nonnucleoside Reverse Transcriptase Inhibitor with Distinct Mutation Development Pathways
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DOI：
10.1128/aac.04201-14
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发表时间：
2015-01-01
影响因子：
4.9
通讯作者：
Lai, Ming-Tain
中科院分区：
文献类型：
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作者：
Feng, Meizhen;Wang, Deping;Lai, Ming-Tain
多拉韦林（DOR，以前称为MK - 1439）是一种1型人类免疫缺陷病毒（HIV - 1）非核苷类逆转录酶抑制剂（NNRTI），目前正处于2b期临床试验阶段。在96孔板中，在低感染复数条件下，用多拉韦林、利匹韦林（RPV）和依非韦伦（EFV）对B亚型病毒（MT4 - 绿色荧光蛋白[GFP]细胞）以及A和C亚型病毒（MT4 - GFP/CCR5细胞）进行了体外耐药性筛选。在含10%胎牛血清的情况下，使用从95%有效浓度（1×EC95）到1000×EC95递增浓度的NNRTI进行耐药性筛选。在用多拉韦林对B亚型病毒进行耐药性筛选时，一种V106A突变病毒导致了两条突变途径，随后分别出现F227L或L234I突变。在对A和C亚型病毒进行耐药性筛选时，检测到了类似的突变发展途径，其中V106A或V106M突变体也是这些途径中的起始病毒。在这项体外耐药性筛选研究中，在B亚型病毒中还鉴定出了在临床环境中通常与利匹韦林和依非韦伦相关的突变，例如分别为E138K和K103N突变体。评估了由多拉韦林、利匹韦林和依非韦伦筛选出的B亚型突变病毒对NNRTI的敏感性。结果表明，多拉韦林筛选出的突变病毒对利匹韦林和依非韦伦敏感，利匹韦林和依非韦伦筛选出的突变体对多拉韦林敏感。当将V106A突变体的复制能力与野生型（WT）病毒进行比较时，突变病毒的适应性比野生型病毒低4倍。
Doravirine (DOR, formerly known as MK-1439) is a human immunodeficiency type 1 virus (HIV-1) nonnucleoside reverse transcriptase inhibitor (NNRTI) that is currently in phase 2b clinical trials. In vitro resistance selection of subtype B virus (MT4-green fluorescent protein [GFP] cells), as well as subtype A and C viruses (MT4-GFP/CCR5 cells) was conducted with DOR, rilpivirine (RPV), and efavirine (EFV) under low-multiplicity-of-infection conditions in a 96-well format. Resistance selection was performed with escalating concentrations of the NNRTIs ranging from the 95% effective concentration (1 x EC95) to 1,000 x EC95 in the presence of 10% fetal bovine serum. In the resistance selection of subtype B virus with DOR, a V106A mutant virus led to two mutation pathways, followed by the emergence separately of either F227L or L234I. In the resistance selection of subtype A and C viruses, similar mutation development pathways were detected, in which a V106A or V106M mutant was also the starting virus in the pathways. Mutations that are commonly associated with RPV and EFV in clinical settings were also identified in subtype B viruses such as the E138K and K103N mutants, respectively, in this in vitro resistance selection study. The susceptibility of subtype B mutant viruses selected by DOR, RPV, and EFV to NNRTIs was evaluated. Results suggest that mutant viruses selected by DOR are susceptible to RPV and EFV and mutants selected by RPV and EFV are susceptible to DOR. When the replication capacity of the V106A mutant was compared with that of the wild-type (WT) virus, the mutant virus was 4-fold less fit than the WT virus.