Evaluating the therapeutic potential of a non-natural nucleotide that inhibits human ribonucleotide reductase.
Evaluating the therapeutic potential of a non-natural nucleotide that inhibits human ribonucleotide reductase.
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DOI：
10.1158/1535-7163.mct-12-0199
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发表时间：
2012-10
影响因子：
5.7
通讯作者：
Dealwis C
中科院分区：
文献类型：
--
作者：
Ahmad MF;Wan Q;Jha S;Motea E;Berdis A;Dealwis C
人核糖核苷酸还原酶 (hRR) 是参与 dNTP 从头合成的关键酶，因此代表了针对过度增殖性疾病（尤其是癌症）的重要治疗靶点。本研究的目的是评估非天然吲哚基-2’-脱氧核苷三磷酸抑制 hRR 活性的能力。这些类似物与 dATP 的结构相似性预测它们将通过结合 hRR 的变构位点来抑制 hRR 活性。计算机分析和体外表征确定了一种特定的类似物，称为 5-硝基-吲哚基-2'-脱氧核糖三磷酸 (5-NITP)，可抑制 hRR。通过等温滴定量热法测定 5-NITP 与 hRR 的结合。测定了与 RR1 结合的 5-NITP 的 X 射线晶体结构。基于细胞的研究证明了相应的非天然核苷对白血病细胞的抗癌作用。 5-NITP 以微摩尔亲和力与 hRR 结合。结合不会像 dATP 那样诱导 hRR1 六聚化，dATP 是 hRR 的天然变构抑制剂，以高亲和力与 A 位点结合。酿酒酵母 RR1-5-NITP (ScRR1-5-NITP) 复合物的 X 射线晶体结构以 2.3 Å 分辨率测定，表明 5-NITP 不与 A 位结合，而是与 S 位结合。无论如何，5-NIdR 通过改变细胞周期进程对人类白血病细胞产生细胞抑制和细胞毒性作用。我们的研究为开发具有改进的 hRR 效力和功效的新型抑制剂提供了有用的见解。
Human ribonucleotide reductase (hRR) is the key enzyme involved in de novo dNTP synthesis and thus represents an important therapeutic target against hyperproliferative diseases, most notably cancer. The purpose of this study was to evaluate the ability of non-natural indolyl-2’-deoxynucleoside triphosphates to inhibit the activity of hRR. The structural similarities of these analogs with dATP predicted that they would inhibit hRR activity by binding to its allosteric sites. In silico analysis and in vitro characterization identified one particular analog designated as 5-nitro-indolyl-2'-deoxyribose triphosphate (5-NITP) that inhibits hRR. 5-NITP binding to hRR was determined by isothermal titration calorimetry. X-ray crystal structure of 5-NITP bound to RR1 was determined. Cell-based studies demonstrated the anti-cancer effects of the corresponding non-natural nucleoside against leukemia cells. 5-NITP binds to hRR with micromolar affinity. Binding does not induce hexamerization of hRR1 like dATP, the native allosteric inhibitor of hRR that binds with high affinity to the A-site. The X-ray crystal structure of S. cerevisiae RR1-5-NITP (ScRR1-5-NITP) complex determined to 2.3 Å resolution shows that 5-NITP does not bind to the A-site but rather at the S-site. Regardless, 5-NIdR produces cytostatic and cytotoxic effects against human leukemia cells by altering cell-cycle progression. Our studies provide useful insights towards developing new inhibitors with improved potency and efficacy against hRR.
影响因子：
4
作者：
Eng, Kevin;Scouten-Ponticelli, Sarah K.;Sutton, Mark;Berdis, Anthony
通讯作者：
Berdis, Anthony