Compound Safety Prediction, Worldwide Medicinal Chemistry, Pfizer Inc., Groton, CT 06340, USA.
Toxicol In Vitro. 2011 Apr;25(3):715-23. doi: 10.1016/j.tiv.2010.12.010. Epub 2010 Dec 22.
Lysosomes are acidic organelles that are essential for the degradation of old organelles and engulfed microbes. Furthermore, lysosomes play a key role in cell death. Lipophilic or amphiphilic compounds with a basic moiety can become protonated and trapped within lysosomes, causing lysosomal dysfunction. Therefore, high-throughput screens to detect lysosomotropism, the accumulation of compounds in lysosomes, are desirable. Hence, we developed a 96-well format, high content screening assay that measures lysosomotropism and cytotoxicity by quantitative image analysis. Forty drugs, including antidepressants, antipsychotics, antiarrhythmics and anticancer agents, were tested for their effects on lysosomotropism and cytotoxicity in H9c2 cells. The assay correctly identified drugs known to cause lysosomotropism and revealed novel information showing that the anticancer drugs, gefitinib, lapatinib, and dasatinib, caused lysosomotropism. Although structurally and pharmacologically diverse, drugs that were lysosomotropic shared certain physicochemical properties, possessing a ClogP>2 and a basic pKa between 6.5 and 11. In contrast, drugs which did not lie in this physicochemical property space were not lysosomotropic. The assay is a robust, rapid screen that can be used to identify lysosomotropic, as well as, cytotoxic compounds, and can be positioned within a screening paradigm to understand the role of lysosomotropism as a contributor to drug-induced toxicity.
溶酶体是酸性细胞器，对于降解衰老的细胞器和吞噬的微生物至关重要。此外，溶酶体在细胞死亡中发挥着关键作用。具有碱性部分的亲脂性或两亲性化合物可以质子化并被困在溶酶体中，导致溶酶体功能障碍。因此，需要进行高通量筛选以检测溶酶体亲嗜性，即化合物在溶酶体中的积累。因此，我们开发了一种 96 孔格式的高通量筛选测定法，通过定量图像分析来测量溶酶体亲嗜性和细胞毒性。该测定法在 H9c2 细胞中测试了 40 种药物（包括抗抑郁药、抗精神病药、抗心律失常药和抗癌药）对溶酶体亲嗜性和细胞毒性的影响。该测定法正确识别了已知引起溶酶体亲嗜性的药物，并揭示了新的信息，表明抗癌药吉非替尼、拉帕替尼和达沙替尼引起了溶酶体亲嗜性。尽管结构和药理学上有差异，但溶酶体亲嗜性的药物具有某些物理化学性质，具有 ClogP>2 和碱性 pKa 在 6.5 到 11 之间。相比之下，不在该物理化学性质空间中的药物则不是溶酶体亲嗜性的。该测定法是一种稳健、快速的筛选方法，可用于识别溶酶体亲嗜性以及细胞毒性化合物，并可在筛选范式中定位，以了解溶酶体亲嗜性作为药物诱导毒性的一个贡献因素的作用。
