Inhibition of HIV-1 Replication by a Bis-Thiadiazolbenzene-1,2-Diamine That Chelates Zinc Ions from Retroviral Nucleocapsid Zinc Fingers
Inhibition of HIV-1 Replication by a Bis-Thiadiazolbenzene-1,2-Diamine That Chelates Zinc Ions from Retroviral Nucleocapsid Zinc Fingers
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DOI：
10.1128/aac.01671-09
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发表时间：
2010-04-01
影响因子：
4.9
通讯作者：
Daelemans, Dirk
中科院分区：
文献类型：
--
作者：
Pannecouque, Christophe;Szafarowicz, Beata;Daelemans, Dirk
人类免疫缺陷病毒1型（HIV-1）核衣壳p7（NCp 7）蛋白含有两个高度保守的“CCHC”锌指结构域，这是病毒复制的几个阶段所需的。碱性残基侧翼的锌指，这两个决定因素都需要高亲和力结合RNA。之前发现几种化合物通过与锌指半胱氨酸残基的巯基反应来靶向NCp 7。在这里，我们已经鉴定了N，N '-双（1，2，3-噻二唑-5-基）苯-1，2-二胺（NV 038），其有效地阻断广谱HIV-1、HIV-2和猿猴免疫缺陷病毒（SIV）株的复制。添加时间实验表明，NV 038干扰病毒进入后但在早期逆转录反应之前或同时的病毒复制周期的一个步骤，表明与核壳蛋白p7相互作用。事实上，在体外，NV 038有效地从NCp 7中消耗锌，这是通过抑制NCp 7诱导的cTAR（TAR的互补DNA序列）的不稳定化来实现的。一个化学模型表明，在这种化合物中的酯的两个羰基氧参与螯合的Zn 2+离子。因此，该化合物通过与先前报道的锌喷射剂不同的机制起作用，因为其结构特征不允许酰基转移到Cys或硫醇-二硫化物交换。这种新的领导和机制研究提供了深入了解未来一代抗NCp 7化合物的设计。
The human immunodeficiency virus type 1 (HIV-1) nucleocapsid p7 (NCp7) protein holds two highly conserved "CCHC" zinc finger domains that are required for several phases of viral replication. Basic residues flank the zinc fingers, and both determinants are required for high-affinity binding to RNA. Several compounds were previously found to target NCp7 by reacting with the sulfhydryl group of cysteine residues from the zinc fingers. Here, we have identified an N,N'-bis(1,2,3-thiadiazol-5-yl)benzene-1,2-diamine (NV038) that efficiently blocks the replication of a wide spectrum of HIV-1, HIV-2, and simian immunodeficiency virus (SIV) strains. Time-of-addition experiments indicate that NV038 interferes with a step of the viral replication cycle following the viral entry but preceding or coinciding with the early reverse transcription reaction, pointing toward an interaction with the nucleocapsid protein p7. In fact, in vitro, NV038 efficiently depletes zinc from NCp7, which is paralleled by the inhibition of the NCp7-induced destabilization of cTAR (complementary DNA sequence of TAR). A chemical model suggests that the two carbonyl oxygens of the esters in this compound are involved in the chelation of the Zn2+ ion. This compound thus acts via a different mechanism than the previously reported zinc ejectors, as its structural features do not allow an acyl transfer to Cys or a thiol-disulfide interchange. This new lead and the mechanistic study presented provide insight into the design of a future generation of anti-NCp7 compounds.