Structural basis for activation of α-boranophosphate nucleotide analogues targeting drug-resistant reverse transcriptase
Structural basis for activation of α-boranophosphate nucleotide analogues targeting drug-resistant reverse transcriptase
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DOI：
10.1093/emboj/19.14.3520
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发表时间：
2000-07-17
期刊：
影响因子：
11.4
通讯作者：
Canard, B
中科院分区：
文献类型：
--
作者：
Meyer, P;Schneider, B;Canard, B
艾滋病化疗受到细胞内活性三磷酸形式核苷类似物浓度不足的限制，导致病毒复制的不完全抑制和耐药病毒的出现。比较了核苷二磷酸激酶对药物的激活作用和对HIV-1逆转录酶的抑制作用。我们合成了在α-磷酸上具有硼（BH 3-）基团的类似物，并发现它们是两种酶的底物。与核苷酸二磷酸激酶的复合物的X射线结构提供了其激活的结构基础。该复合物与d4 T三磷酸显示分子内CH。O键有助于催化，胸苷α-硼三磷酸的R-p非对映异构体像正常底物一样结合。使用临床相关化合物AZT和d4 T的α-（R-p）-硼烷磷酸衍生物，α-硼烷基团的存在改善了核苷酸二磷酸激酶的磷酸化和逆转录的抑制。此外，修复阻断的DNA链焦磷酸解显着减少在耐药病毒中发现的变异逆转录酶轴承取代。因此，靶向逆转录酶的类似物的α-硼烷修饰可能在对抗病毒耐药性方面具有通用价值。
AIDS chemotherapy is limited by inadequate intracellular concentrations of the active triphosphate form of nucleoside analogues, leading to incomplete inhibition of viral replication and the appearance of drug-resistant virus. Drug activation by nucleoside diphosphate kinase and inhibition of HIV-1 reverse transcriptase were studied comparatively. We synthesized analogues with a borano (BH3-) group on the alpha-phosphate, and found that they are substrates for both enzymes. X-ray structures of complexes with nucleotide diphosphate kinase provided a structural basis for their activation. The complex: with d4T triphosphate displayed an intramolecular CH...O bond contributing to catalysis, and the R-p diastereoisomer of thymidine alpha-boranotriphosphate bound like a normal substrate. Using alpha-(R-p)-boranophosphate derivatives of the clinically relevant compounds AZT and d4T, the presence of the alpha-borano group improved both phosphorylation by nucleotide diphosphate kinase and inhibition of reverse transcription. Moreover, repair of blocked DNA chains by pyrophosphorolysis was reduced significantly in variant reverse transcriptases bearing substitutions found in drug-resistant viruses. Thus, the alpha-borano modification of analogues targeting reverse transcriptase may be of generic value in fighting viral drug resistance.