Styrylquinolines, integrase inhibitors acting prior to integration:: a new mechanism of action for anti-integrase agents
Styrylquinolines, integrase inhibitors acting prior to integration:: a new mechanism of action for anti-integrase agents
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DOI：
10.1128/jvi.78.11.5728-5736.2004
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发表时间：
2004-06-01
影响因子：
5.4
通讯作者：
Leh, H
中科院分区：
文献类型：
--
作者：
Bonnenfant, S;Thomas, CM;Leh, H
我们之前已经证明苯乙烯喹啉（sql）在体外是整合酶抑制剂。它们与长末端重复底物竞争整合酶。在这里，我们描述了这些分子的细胞作用模式。我们证明sql不会干扰病毒进入。事实上，在体外实验中，高达50%抑制浓度20倍的浓度并不会抑制细胞间融合，也不会影响GP120与CD4之间的相互作用。此外，逆转录病毒包膜的伪型不影响药物活性。定量反转录PCR实验表明，sql不抑制基因组RNA的进入。相比之下，用sql治疗人类免疫缺陷病毒i型感染的细胞减少了晚期cDNA的数量，这首次表明整合酶靶向分子可能影响逆转录过程中DNA的积累。当病毒在sql浓度增加的情况下生长时，整合酶基因出现突变，证实了sql的细胞靶标。最后，当将这些突变引入野生型序列时，这些突变导致了sql抗性病毒。相反，sql对逆转录酶抑制剂和二酮酸抗性病毒具有完全活性，将sql定位为第二组抗整合酶化合物。
We have previously shown that styrylquinolines (SQLs) are integrase inhibitors in vitro. They compete with the long terminal repeat substrate for integrase. Here, we describe the cellular mode of action of these molecules. We show that SQLs do not interfere with virus entry. In fact, concentrations of up to 20 times the 50% inhibitory concentration did not inhibit cell-to-cell fusion or affect the interaction between GP120 and CD4 in vitro. Moreover, the pseudotype of the retrovirus envelope did not affect drug activity. Quantitative reverse transcription PCR experiments showed that SQLs do not inhibit the entry of the genomic RNA. In contrast, the treatment of human immunodeficiency virus type I-infected cells with SQLs reduced the amount of the late cDNA, suggesting for the first time that integrase targeting molecules may affect the accumulation of DNA during reverse transcription. The cellular target of SQLs was confirmed by the appearance of mutations in the integrase gene when viruses were grown in the presence of increasing concentrations of SQLs. Finally, these mutations led to SQL-resistant viruses when introduced into the wild-type sequence. In contrast, SQLs were fully active against reverse transcriptase inhibitor- and diketo acid-resistant viruses, positioning SQLs as a second group of anti-integrase compounds.