We recently reported 2,4,5-trimethylpyridin-3-ol with C(6)-azacyclonol, whose code name is BJ-1207, showing a promising anticancer activity by inhibiting NOX-derived ROS in A549 human lung cancer cells. The present study was focused on structural modification of the azacyclonol moiety of BJ-1207 to find a compound with better anticancer activity. Ten new compounds (3A–3J) were prepared and evaluated their inhibitory actions against proliferation of eighteen cancer cell lines as a primary screening. Among the ten derivatives of BJ-1207, the effects of compounds 3A and 3J on DU145 and PC-3, androgen-refractory cancer cell lines (ARPC), were greater than the parent compound, and compound 3A showed better activity than 3J. Antitumor activity of compound 3A was also observed in DU145-xenografted chorioallantoic membrane (CAM) tumor model. In addition, the ligand-based target prediction and molecular docking study using DeepZema® server showed compound 3A was a ligand to M3 muscarinic acetylcholine receptor (M3R) which is overexpressed in ARPC. Carbachol, a muscarinic receptor agonist, concentration dependently increased proliferation of DU145 in the absence of serum, and it also activated NADPH oxidase (NOX). The carbachol-induced proliferation and NOX activity was significantly blocked by compounds 3A in a concentration-dependent manner. This finding might become a new milestone in the development of pyridinol-based anti-cancer agents against ARPC.

我们最近报道了 2,4,5-trimethylpyridin-3-ol 和 C(6)-azacyclonol，其代号为 BJ-1207，通过抑制 A549 人肺癌细胞中 NOX 衍生的 ROS 显示出有希望的抗癌活性。本研究的重点是对 BJ-1207 的氮杂环醇部分进行结构修饰，以寻找具有更好抗癌活性的化合物。制备了 10 种新化合物 ( 3A – 3J)，并评估了它们对 18 种癌细胞系增殖的抑制作用，作为初步筛选。在BJ-1207的10种衍生物中，化合物3A和3J对DU145和PC-3、雄激素难治性癌细胞系（ARPC）的影响大于母体化合物和化合物3A显示出比3J更好的活性。在 DU145 异种移植绒毛膜尿囊膜 (CAM) 肿瘤模型中也观察到化合物3A 的抗肿瘤活性。此外，使用 DeepZema® 服务器进行的基于配体的靶标预测和分子对接研究表明，化合物3A是 M3 毒蕈碱乙酰胆碱受体 (M3R) 的配体，该受体在 ARPC 中过度表达。Carbachol 是一种毒蕈碱受体激动剂，在没有血清的情况下，浓度依赖性地增加 DU145 的增殖，并且它还激活 NADPH 氧化酶 (NOX)。化合物3A显着阻断了碳酰胆碱诱导的增殖和NOX活性以浓度依赖的方式。这一发现可能成为开发基于吡啶醇的抗 ARPC 抗癌药物的新里程碑。