含咔唑和吡唑的抗癌化合物的潜在靶点和作用机制。
The Potential Targets and Mechanisms of a Carbazole and Pyrazole Containing Anticancer Compound.
机构信息
Vanderbilt University Institute of Imaging Science, Vanderbilt University, Nashville, TN 37232, United States.
Department of Radiology and Radiological Sciences, Vanderbilt University, Nashville, TN 37232, United States.
出版信息
Curr Cancer Drug Targets. 2020;20(5):364-371. doi: 10.2174/1568009620666200115162343.
AIMS
Characterization of a small anticancer compound.
BACKGROUND
The development of small molecules as new anti-cancer therapeutics is necessary to improve anti-tumor efficacy and reduce toxicities, especially for the treatment of brain tumors, where only small molecules can effectively cross the brain-blood barrier. Several novel hits were previously selected by concurrently screening colon and glioma cancer cell lines with a sensorconjugated reporter system. Here, we focused on one of them.
OBJECTIVE
Elucidating the potential target(s) of a novel anticancer compound.
METHODS
Computer-assisted structural and motif analysis (least absolute shrinkage and selection operator or LASSO score) was used to assess compound's targets, then direct kinase activity assays were used for the confirmation; Western blot of phosphorylated kinases, as well as FACS and caspase 3/7 activity assays, were used to decipher the action mechanisms. Finally, the expression profiling of proteins involved in various G-protein pathways by real-time PCR was performed.
RESULTS
The small chemical, (4E)-4-[2-(9-ethyl-9H-carbazol-3-yl)hydrazin-1-ylidene]-3-methyl- 4,5-dihydro-1H-pyrazol-5-one, with a formula C18H17N5O and MW of 319.36, designated as VUGX01, was predicted to be a ligand/inhibitor to receptor tyrosine kinases (RTKs) by computer analysis (least absolute shrinkage and selection operator or LASSO score). However, direct analysis with recombinant kinases showed that it is not an effective inhibitor to the popular receptor kinases at 1μM concentration. This compound can activate caspases in some tumor cell lines but has minimal effects on the cell cycle. Drug treatments lead to the changes in phosphorylation of AKT and c- RAF, as well as the expression level of MAP2K, suggesting this compound may interact with Gprotein coupled receptors (GPCRs). The expression profiling of 82 proteins involved in various Gprotein pathways by real-time PCR showed that the treatment up-regulates the expression of several proteins, including angiotensinogen, angiotensin II receptor, and IP3-kinase catalytic subunit gamma.
CONCLUSION
VUGX01 can effectively block proliferation and induce apoptosis of certain types of cancer cells, even it is predicted by high LASSO score, but it is not an effective RTKs inhibitor, it may inhibit cell growth through acting as a novel ligand to one or several GPCRs.
目的
对一种小分子抗癌化合物进行表征。
背景
开发小分子作为新的抗癌治疗药物对于提高抗肿瘤疗效和降低毒性是必要的，特别是对于治疗脑肿瘤，只有小分子才能有效地穿过血脑屏障。以前通过与传感器偶联报告系统同时筛选结肠和神经胶质瘤癌细胞系，选择了几种新型化合物。在此，我们专注于其中一种。
目的
阐明新型抗癌化合物的潜在靶标。
方法
计算机辅助结构和基序分析（最小绝对收缩和选择算子或 LASSO 评分）用于评估化合物的靶标，然后直接进行激酶活性测定进行确认；磷酸化激酶的 Western blot 以及 FACS 和 caspase 3/7 活性测定用于破译作用机制。最后，通过实时 PCR 进行涉及各种 G 蛋白途径的蛋白质表达谱分析。
结果
小分子化合物（4E）-4-[2-（9-乙基-9H-咔唑-3-基）腙-1-基]-3-甲基-4,5-二氢-1H-吡唑-5-酮，化学式为 C18H17N5O，MW 为 319.36，命名为 VUGX01，通过计算机分析（最小绝对收缩和选择算子或 LASSO 评分）预测为受体酪氨酸激酶（RTKs）的配体/抑制剂。然而，在 1μM 浓度下，用重组激酶进行直接分析表明，它不是一种有效的流行受体激酶抑制剂。该化合物可在一些肿瘤细胞系中激活半胱天冬酶，但对细胞周期影响较小。药物处理导致 AKT 和 c-RAF 的磷酸化以及 MAP2K 的表达水平发生变化，表明该化合物可能与 G 蛋白偶联受体（GPCRs）相互作用。通过实时 PCR 对涉及各种 G 蛋白途径的 82 种蛋白质的表达谱进行分析表明，该处理可上调几种蛋白质的表达，包括血管紧张素原、血管紧张素 II 受体和 IP3-激酶催化亚基 γ。
结论
VUGX01 可有效阻断某些类型癌细胞的增殖并诱导其凋亡，尽管其 LASSO 评分较高，但它不是有效的 RTKs 抑制剂，它可能通过作为一种新型配体作用于一种或几种 GPCR 来抑制细胞生长。
Zotero 插件