Isolation of two highly potent and non-toxic inhibitors of human immunodeficiency virus type 1 (HIV-1) integrase from Salvia miltiorrhiza
Isolation of two highly potent and non-toxic inhibitors of human immunodeficiency virus type 1 (HIV-1) integrase from Salvia miltiorrhiza
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DOI：
10.1016/s0166-3542(02)00011-6
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发表时间：
2002-07-01
影响因子：
7.6
通讯作者：
Huang, RCC
中科院分区：
文献类型：
--
作者：
Abd-Elazem, IS;Chen, HS;Huang, RCC
丹参水溶性提取物在体外对HIV-1整合酶活性和体内病毒复制均有明显抑制作用。我们开发了一种广泛的纯化方案，利用整合酶的3'加工活性作为纯化指导和测定，分离出有效的、无毒的针对人类免疫缺陷病毒1型（HIV-1）的抑制剂。从丹参根中分离得到两种水溶性化合物M，22和M，32，经核磁共振波谱分析，其纯度为bb0 ~ 99.5%，产率分别为0.018和0.038%。结构鉴定表明M(5)22为石精酸，M(5)32为石精酸b。这两种结构相关的化合物是有效的抗hiv抑制剂，高浓度时对H9细胞无细胞毒性(M（5)22为CC100 bbb297 muM， M(5)32为> 223 muM）。M(5)22和M(5)32抑制HIV-1整合酶3'加工的IC50分别为0.83和0.48 muM。此外，M(5)22和M(5)32抑制了HIV-1整合酶3'-连接目标DNA的催化活性，M(5)22和M(5)32的IC50分别为0.48和0.37 muM。此外，动力学和机制研究表明，药物与HIV-1整合酶结合并抑制酶活性的速度很快。M(5)22和M(5)32都不能阻止HIV进入H9细胞。它们对感染细胞的逆转录酶活性也没有抑制作用。细胞内强阻断和全长病毒DNA水平在药物治疗后保持不变。然而，这两种抑制剂对H9细胞的急性HIV-1感染均有较强的抑制作用，M(5)22和M(5)32的IC50值分别为2和6.9 muM。因此，这两种选择性整合酶抑制剂因其高效力和无细胞毒性而有望成为一类新的艾滋病治疗药物。(c) 2002 Elsevier Science B.V.版权所有
Water soluble extracts of the herbal plant, Salvia miltiorrhiza (Danshen) exhibited potent effect against HIV-1 integrase activity in vitro and viral replication in vivo. We have developed an extensive purification scheme to isolate effective, non-toxic inhibitors against human immunodeficiency virus type 1 (HIV-1) using the 3'-processing activity of integrase as a purification guide and assay. Two water soluble compounds, M,22 and M,32, have been discovered by isolating them from S. miltiorrhiza roots in purities of >99.5%, as shown by NMR spectral analysis with yields of 0.018 and 0.038%, respectively. Structural determination revealed that M(5)22 is lithospermic acid and M(5)32 is lithospermic acid B. These two structurally related compounds are potent anti-HIV inhibitors and showed no cytotoxicity to H9 cells at high concentrations (CC100 > 297 muM for M(5)22 and > 223 muM for M(5)32). The IC50 for inhibition of 3'-processing by HIV-1 integrase was found to be 0.83 muM for M(5)22 and 0.48 muM for M(5)32. In addition, M(5)22 and M(5)32 inhibited HIV-1 integrase catalytic activities of 3'-joining to the target DNA with IC50 of 0.48 muM for M(5)22 and 0.37 muM for M(5)32. Furthermore, kinetic and mechanistic studies suggested that drug binding to HIV-1 integrase and inhibition of enzymatic activity occur at a fast rate. Both M(5)22 and M(5)32 do not prevent HIV entry in H9 cells. They also show no inhibition of reverse transcriptase activity in infected cells. The levels of intracellular strong stop and full-length viral DNA remained unchanged following drug treatment. However, both inhibitors strongly suppressed the acute HIV-1 infection of H9 cells with IC50 values of 2 and 6.9 muM for M(5)22 and M(5)32, respectively. Thus these two selective integrase inhibitors hold promise as a novel class of therapeutic drugs for AIDS based on their high potencies and absence of cytotoxicity. (c) 2002 Elsevier Science B.V. All rights reserved.