DEB025 (Alisporivir) inhibits hepatitis C virus replication by preventing a cyclophilin A induced cis-trans isomerisation in domain II of NS5A.
DEB025 (Alisporivir) inhibits hepatitis C virus replication by preventing a cyclophilin A induced cis-trans isomerisation in domain II of NS5A.
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DOI：
10.1371/journal.pone.0013687
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发表时间：
2010-10-27
期刊：
影响因子：
3.7
通讯作者：
Neyts J
中科院分区：
文献类型：
--
作者：
Coelmont L;Hanoulle X;Chatterji U;Berger C;Snoeck J;Bobardt M;Lim P;Vliegen I;Paeshuyse J;Vuagniaux G;Vandamme AM;Bartenschlager R;Gallay P;Lippens G;Neyts J
DEB025/Debio 025 （Alisporivir）是一种亲环蛋白（Cyp）结合分子，在体内和体外均具有抗丙型肝炎病毒（HCV）的活性。目前正在II期临床试验中进行评估。DEB025与CypA结合，CypA是一种肽基脯氨酸顺式反式异构酶，是HCV复制的重要辅助因子。在这里，我们报告说，选择DEB025的耐药复制子（基因型1b）非常困难，平均需要20周（4个独立实验），而蛋白酶或聚合酶抑制剂通常不到2周。这表明DEB025对抗性具有较高的遗传屏障。NS5A中的突变D320E是复制子基因组中唯一一致选择的突变。仅这一突变就产生了低水平（3.9倍）的耐药性。用DEB025res复制子的相应序列替换野生型（WT）基因组中的NS5A基因（而不是NS5B基因）导致抗性转移。观察到与环孢素A （CsA）的交叉抗性，而NS3蛋白酶和NS5B聚合酶抑制剂对DEB025res复制子保持wt活性。与WT不同，DEB025res复制子在CypA敲除细胞中有效复制。然而，无论NS5A蛋白是来自WT还是DEB025res复制子，DEB025都破坏了CypA与NS5A的相互作用。从NS5A的WT或DEB025res结构域II衍生的肽的核磁共振滴定实验以定量的方式证实了这一观察结果。有趣的是，对含有D320或E320的两种20-mer NS5A肽的比较核磁共振研究显示，主要构象和次要构象之间的种群发生了变化。这些数据表明，D320E可能通过减少NS5A对cypa依赖性异构化的需要而赋予对DEB025的低水平耐药性。长期的DEB025治疗和多种基因型改变可能是产生对DEB025的显著抗性所必需的，这是抗性高屏障的基础。
DEB025/Debio 025 (Alisporivir) is a cyclophilin (Cyp)-binding molecule with potent anti-hepatitis C virus (HCV) activity both in vitro and in vivo. It is currently being evaluated in phase II clinical trials. DEB025 binds to CypA, a peptidyl-prolyl cis-trans isomerase which is a crucial cofactor for HCV replication. Here we report that it was very difficult to select resistant replicons (genotype 1b) to DEB025, requiring an average of 20 weeks (four independent experiments), compared to the typically <2 weeks with protease or polymerase inhibitors. This indicates a high genetic barrier to resistance for DEB025. Mutation D320E in NS5A was the only mutation consistently selected in the replicon genome. This mutation alone conferred a low-level (3.9-fold) resistance. Replacing the NS5A gene (but not the NS5B gene) from the wild type (WT) genome with the corresponding sequence from the DEB025res replicon resulted in transfer of resistance. Cross-resistance with cyclosporine A (CsA) was observed, whereas NS3 protease and NS5B polymerase inhibitors retained WT-activity against DEB025res replicons. Unlike WT, DEB025res replicon replicated efficiently in CypA knock down cells. However, DEB025 disrupted the interaction between CypA and NS5A regardless of whether the NS5A protein was derived from WT or DEB025res replicon. NMR titration experiments with peptides derived from the WT or the DEB025res domain II of NS5A corroborated this observation in a quantitative manner. Interestingly, comparative NMR studies on two 20-mer NS5A peptides that contain D320 or E320 revealed a shift in population between the major and minor conformers. These data suggest that D320E conferred low-level resistance to DEB025 probably by reducing the need for CypA-dependent isomerisation of NS5A. Prolonged DEB025 treatment and multiple genotypic changes may be necessary to generate significant resistance to DEB025, underlying the high barrier to resistance.
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影响因子：
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