CA-5f is a potent late-stage macroautophagy/autophagy inhibitor via inhibiting autophagosome-lysosome fusion.

对人类脐静脉内皮细胞(HUVECs)使用CA-5f处理1小时后，能够降低细胞骨架蛋白和膜运输蛋白的水平。CA-5f对A549非小细胞肺癌(NSCLC)细胞表现出强烈的细胞毒性，但对正常的HUVECs的细胞毒性较低，其机制是通过增加线粒体来源的活性氧(ROS)产量。

CA-5f 在体内单独使用时有效抑制了A549肺癌异种移植瘤的生长，并展现出了极佳的耐受性。Western blot、免疫荧光以及TdT介导的dUTP末端标记(TUNEL)实验的结果显示，CA-5f抑制了自噬流，诱导了凋亡，并且未影响CTSb (cathepsin B)和CTSD (cathepsin D)的水平，这些结果与体外数据一致。

A549 cells or HUVECs were seeded into 100-mm culture dishes. When cells reached 80% confluency, they were exposed to DMSO or CA-5f for 6 h or 24 h. Then the cells were fixed, dehydrated, embedded, sectioned, and stained as previously described. Finally, the ultrathin sections of these samples were observed under a JEM-1230 transmission electron microscopy.

Four-week-old BALB/c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Lt and kept in specific pathogen-free conditions at the Experimental Animal Center in the Institute of Medicine in Zhengzhou University. Nude mice underwent subcutaneous injection of A549 cells (1 × 10^6) at the right scapula. 8 days after A549 cell implantation, mice exhibited palpable tumors, then they were randomly divided into 2 groups (each group n = 5). CA-5f was injected via caudal vein every 2 days for up to 30 days. DMSO was used as a control. From the initial injection, the tumor size was measured every 2 days. Tumor volumes were calculated as tumor length × (square of width)/2. At the end of the experiment, all mice were sacrificed and the tumors were resected immediately and photographed. The weight of each tumor was measured.