School of Pharmaceutical Sciences, Nanjing Technical University, No. 5 Xinmofan Road, Nanjing 210009, People's Republic of China.
Med Chem. 2022;18(6):679-693. doi: 10.2174/1573406418666211210163817.
NEDD8 (neural precursor cell expressed developmentally downregulated protein 8) is one of the ubiquitin-like proteins which is activated by the NEDD8 activating enzyme (NAE). The overexpressed NAE can cause a variety of diseases such as numerous cancer types and inflammatory diseases. The selective inhibition of NAE could mediate the rate of ubiquitination and the subsequent degradation of proteins associated with cancer so as to achieve the purpose of treatment.
In this article, we decided to study the synthesis and screening of coumarin scaffold derivatives against cancer cell lines, specifically the human pancreatic cancer cell line BxPC-3.
Twenty-four targeted compounds were synthesized, and their anti-proliferative activity against three cancer cell lines, cytotoxicity against three normal cell lines through CCK-8 and MTT assay were evaluated to screen out the candidate compound. Then the target was further confirmed by both enzyme and cell-based experiments, as well as cell apoptosis research.
Several new 4-position substituted coumarin derivatives (12a~x) were synthesized and most of them exhibit antiproliferative activity in three cancer cell lines. A series of experiments were performed to identify the best candidate compound 12v. This compound displayed the highest potency against BxPC-3 with an IC50 value of 0.28 μM. It can also inhibit NAE activity in enzyme and cellbased assay, and induce CRLs-mediated accumulation of the substrate and apoptosis in BxPC-3 cells. Meanwhile, it exhibited relatively low toxicity in three normal cells.
Based on these results, we found that compound 12v inhibited NAE activity in enzyme and cell-based systems and induced apoptosis in BxPC-3 cells. Additionally, it also had a low toxicity. These results suggested that 12v may be promising lead compounds for the development of new anticancer drugs.
NEDD8（神经前体细胞表达发育下调蛋白 8）是一种泛素样蛋白，可被 NEDD8 激活酶（NAE）激活。NAE 的过表达可导致多种疾病，如多种癌症类型和炎症性疾病。NAE 的选择性抑制可以调节与癌症相关的蛋白质的泛素化和随后的降解速度，从而达到治疗的目的。
本文旨在研究香豆素骨架衍生物对癌细胞系，特别是人胰腺癌细胞系 BxPC-3 的合成和筛选。
合成了 24 个靶向化合物，并通过 CCK-8 和 MTT 测定法评估它们对三种癌细胞系的抗增殖活性和对三种正常细胞系的细胞毒性，以筛选出候选化合物。然后通过酶和基于细胞的实验以及细胞凋亡研究进一步确认靶标。
合成了几种新的 4-位取代香豆素衍生物（12a~x），它们大多数对三种癌细胞系均具有抗增殖活性。进行了一系列实验以鉴定最佳候选化合物 12v。该化合物对 BxPC-3 的抑制作用最强，IC50 值为 0.28 μM。它还可以抑制酶和基于细胞的测定中的 NAE 活性，并诱导 CRLs 介导的底物积累和 BxPC-3 细胞凋亡。同时，它在三种正常细胞中表现出相对较低的毒性。
基于这些结果，我们发现化合物 12v 抑制了 NAE 在酶和基于细胞的系统中的活性，并诱导了 BxPC-3 细胞凋亡。此外，它的毒性也较低。这些结果表明，12v 可能是开发新型抗癌药物的有前途的先导化合物。
