Schneider D J, Feigon J, Hostomsky Z, Gold L
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder 80309, USA.
Biochemistry. 1995 Jul 25;34(29):9599-610. doi: 10.1021/bi00029a037.
The reverse transcriptase (RT) of HIV-1 is a plausible target for therapeutic agents aimed at inhibiting propagation of the virus. We have used "irrational drug design", that is, combinatorial chemistry with oligonucleotide libraries, to identify high-affinity ligands aimed at HIV-1 RT. The methodology, termed SELEX (systematic evolution of ligands by exponential enrichment), was employed with a single-stranded DNA library. The selected ssDNA ligands bind HIV-1 RT with Kd values as low as 1 nM and inhibit the RNA-dependent DNA-polymerase activity of the enzyme with Ki values as low as 0.3 nM. We also demonstrate the high specificity of one ligand able to selectively discriminate between the reverse transcriptases of HIV-1, AMV, and MMLV. These ssDNA molecules may be useful as inhibitors or as models for the design of small molecule inhibitors of HIV-1 RT in vivo.
人类免疫缺陷病毒1型（HIV-1）的逆转录酶（RT）是旨在抑制该病毒传播的治疗药物的一个合理靶点。我们采用了“非理性药物设计”，即利用寡核苷酸文库进行组合化学，来鉴定针对HIV-1 RT的高亲和力配体。这种方法被称为SELEX（指数富集配体的系统进化），使用的是单链DNA文库。所选择的单链DNA配体与HIV-1 RT结合的解离常数（Kd）低至1 nM，并以低至0.3 nM的抑制常数（Ki）抑制该酶的RNA依赖性DNA聚合酶活性。我们还证明了一种配体具有高度特异性，能够在HIV-1、禽成髓细胞瘤病毒（AMV）和莫洛尼氏鼠白血病病毒（MMLV）的逆转录酶之间进行选择性区分。这些单链DNA分子可能作为抑制剂，或作为在体内设计HIV-1 RT小分子抑制剂的模型。
