Background: NEDD8 activating enzyme (NAE) plays a critical role in various cellular functions in carcinomas. 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.

Objective: In this article, we decided to study the synthesis and screening of 4-amino substituted 2H-chromen-2-one derivatives against cancer cell lines, specifically the human pancreatic cancer cell line BxPC-3.

Methods: After synthesis of twenty targeted compounds, we evaluated their anti-proliferative activity against six cancer cell lines, cytotoxicity against three normal cell lines through MTT assay, and hemolysis to screen out the candidate compound, which was further conducted drug-like physical property measurement, target confirmation by enzyme-based experiment, cell apoptosis, and synergistic effect research.

Results: Starting from intermediates 4 and 5, several new 4-amino substituted 2H-chromen-2-one derivatives (9-28) were synthesized and evaluated for their cell activities using six cancer cell lines. We performed tests of cytotoxicity, hemolysis, ATP-dependent NAE inhibition in the enzyme- based system, apoptosis, and synergistic effect in BxPC-3 cells against the best candidate compound 21.

Conclusion: Based on these results, we found that compound 21 inhibited NAE activity in an ATP-dependent manner in the enzyme-based system, induced apoptosis in BxPC-3 cells, and synergized with bortezomib on BxPC-3 cell growth inhibition. Additionally, it had low toxicity with reasonable Log P-value and water solubility.

背景：NEDD8活化酶（NAE）在癌症的各种细胞功能中起着至关重要的作用。NAE的选择性抑制可以介导泛素化的速度以及随后与癌症相关的蛋白质的降解，从而达到治疗目的。

目的：在本文中，我们决定研究针对癌细胞系（特别是人胰腺癌细胞系BxPC-3）的4-氨基取代的2H-chromen-2-one衍生物的合成和筛选。

方法：合成20种目标化合物后，我们通过MTT分析评估了它们对六种癌细胞系的抗增殖活性，对三种正常细胞系的细胞毒性，并通过溶血作用筛选出候选化合物，从而进一步进行了类药物的物理性质测量，通过基于酶的实验确定靶标，细胞凋亡以及协同效应研究。

结果：从中间体4和5开始，合成了几种新的4-氨基取代的2H-chromen-2-one衍生物（9-28），并使用6种癌细胞系评估了它们的细胞活性。我们进行了细胞毒性，溶血，基于酶的系统中ATP依赖性NAE抑制，细胞凋亡以及BxPC-3细胞针对最佳候选化合物21的协同效应的测试。

结论：基于这些结果，我们发现化合物21在基于酶的系统中以ATP依赖性方式抑制NAE活性，诱导BxPC-3细胞凋亡，并与硼替佐米协同抑制BxPC-3细胞的生长。此外，它具有低毒性，具有合理的Log P值和水溶性。