Discovery of inhibitors that elucidate the role of UCH-L1 activity in the H1299 lung cancer cell line
Discovery of inhibitors that elucidate the role of UCH-L1 activity in the H1299 lung cancer cell line
复制标题
DOI：
10.1016/j.chembiol.2003.08.010
复制
发表时间：
2003-09-01
影响因子：
--
通讯作者：
Lansbury, PT
中科院分区：
文献类型：
--
作者：
Liu, YC;Lashuel, HA;Lansbury, PT
神经元泛素 C 末端水解酶 (UCH-L1) 与帕金森病 (PD)、某些非神经元肿瘤的进展和神经性疼痛有关。某些肺肿瘤来源的细胞系表达 UCH-L1，但在正常肺组织中不表达，这表明这种酶在肿瘤进展中发挥着作用，无论是作为触发因素还是作为反应。 UCH-L1 的小分子抑制剂将有助于区分这些情况。通过利用高通量筛选 (HTS) 寻找抑制剂和传统药物化学来优化其亲和力和特异性，我们鉴定了一类靛红 O-酰基肟，与其全身同种型 UCH-L3 相比，它可以选择性抑制 UCH-L1。该类别的三个代表（30、50、51）对于 UCH-L1 的 IC50 值为 0.80-0.94 μM，对于 UCH-L3 的 IC50 值为 17-25 μM。 UCH-L1 的 K-i 为 30 为 0.40 muM，抑制是可逆的、竞争性的和活性位点定向的。两种靛红肟抑制剂增加了 H1299 肺肿瘤细胞系的增殖，但对不表达 UCH-L1 的肺肿瘤细胞系没有影响。使用 RNAi 抑制 H1299 细胞系中的 UCH-L1 表达具有类似的增殖作用，表明 UCH-L1 酶活性具有抗增殖作用，并且 UCH-L1 表达可能是对肿瘤生长的反应。这种反应的分子机制仍有待确定。
Neuronal ubiquitin C-terminal hydrolase (UCH-L1) has been linked to Parkinson's disease (PD), the progression of certain nonneuronal tumors, and neuropathic pain. Certain lung tumor-derived cell lines express UCH-L1 but it is not expressed in normal lung tissue, suggesting that this enzyme plays a role in tumor progression, either as a trigger or as a response. Small-molecule inhibitors of UCH-L1 would be helpful in distinguishing between these scenarios. By utilizing high-throughput screening (HTS) to find inhibitors and traditional medicinal chemistry to optimize their affinity and specificity, we have identified a class of isatin O-acyl oximes that selectively inhibit UCH-L1 as compared to its systemic isoform, UCH-L3. Three representatives of this class (30, 50, 51) have IC50 values of 0.80-0.94 muM for UCH-L1 and 17-25 muM for UCH-L3. The K-i of 30 toward UCH-L1 is 0.40 muM and inhibition is reversible, competitive, and active site directed. Two isatin oxime inhibitors increased proliferation of the H1299 lung tumor cell line but had no effect on a lung tumor line that does not express UCH-L1. Inhibition of UCH-L1 expression in the H1299 cell line using RNAi had a similar proproliferative effect, suggesting that the UCH-L1 enzymatic activity is antiproliferative and that UCH-L1 expression may be a response to tumor growth. The molecular mechanism of this response remains to be determined.