DEB025 (Alisporivir) Inhibits Hepatitis C Virus Replication by Preventing a Cyclophilin A Induced Cis-Trans Isomerisation in Domain II of NS5A

被引:143
作者
Coelmont, Lotte [1 ]
Hanoulle, Xavier [2 ]
Chatterji, Udayan [3 ]
Berger, Carola [4 ]
Snoeck, Joke [1 ]
Bobardt, Michael [3 ]
Lim, Precious [3 ]
Vliegen, Inge [1 ]
Paeshuyse, Jan [1 ]
Vuagniaux, Gregoire [5 ]
Vandamme, Anne-Mieke [1 ]
Bartenschlager, Ralf [4 ]
Gallay, Philippe [3 ]
Lippens, Guy [2 ]
Neyts, Johan [1 ]
机构
[1] Katholieke Univ Leuven, Dept Microbiol & Immunol, Rega Inst Med Res, Louvain, Belgium
[2] Univ Lille 1, Ctr Natl Rech Sci, Unite Mixte Rech 8576, F-59655 Villeneuve Dascq, France
[3] Scripps Res Inst, Dept Immunol & Microbial Sci, La Jolla, CA 92037 USA
[4] Heidelberg Univ, Dept Mol Virol, Heidelberg, Germany
[5] Debiopharm, Pharmacol & Screening Unit, Lausanne, Switzerland
来源
PLOS ONE | 2010年 / 5卷 / 10期
关键词
NONSTRUCTURAL PROTEIN 5A; CYCLOSPORINE-A; ISOMERASE ACTIVITY; RNA REPLICATION; RESISTANCE; BINDING; DEBIO-025; IDENTIFICATION; COMBINATION; POLYMERASE;
D O I
10.1371/journal.pone.0013687
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 DEB025(res) 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 DEB025(res) replicons. Unlike WT, DEB025(res) 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 DEB025(res) replicon. NMR titration experiments with peptides derived from the WT or the DEB025(res) 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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页数:14
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