Faculty of Biological Sciences, University of Leeds, Leeds, West Yorkshire LS2 9JT, United Kingdom.
Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, Cambridge, United Kingdom.
Toxicol Appl Pharmacol. 2020 Dec 1;408:115279. doi: 10.1016/j.taap.2020.115279. Epub 2020 Oct 14.
High-throughput, automation-friendly and therapeutically-predictive assays are needed in early drug discovery in order to prioritise compounds and reduce the risk of new drugs causing Drug-Induced Liver Injury (DILI). We evaluated the suitability of high-throughput 3D liver spheroid models of HepG2 (C3A clone) and HepaRG cell lines to predict DILI in early drug development. Spheroids were formed in 384-well ultra-low-attachment plates and dosed via direct acoustic droplet ejection at nine half-log spaced concentrations per compound. Spheroid viability was quantified with an ATP endpoint after a 4-day incubation with 150 drugs with known DILI liability. We derived a margin of safety for each cell line defined as the ratio between the IC values generated for each compound to their maximum plasma concentration C which resulted in optimal classification accuracy. The margin of safety can be used to estimate a maximum safe C for compounds in early drug discovery for which C is not yet known. Both cell lines had similar level of accuracy in predicting DILI, with HepG2 spheroids being more sensitive. HepG2 spheroids had a sensitivity of 58% and a specificity of 83%, while HepaRG spheroids had a sensitivity of 47% and specificity of 86%. Ninety-nine of the 150 compounds were used to compare the relative sensitivities of HepG2 and HepaRG spheroids. HepaRG spheroids were more sensitive to 7 compounds and HepG2 spheroids were more sensitive to 34 compounds. In conclusion, across a diverse group of drugs HepG2 spheroids were more predictive of DILI compared to HepaRG spheroids.
高通量、自动化友好和具有治疗预测性的测定法在早期药物发现中是必要的，以便对化合物进行优先排序，并降低新药导致药物性肝损伤（DILI）的风险。我们评估了 HepG2（C3A 克隆）和 HepaRG 细胞系的高通量 3D 肝球体模型在早期药物开发中预测 DILI 的适用性。球体在 384 孔超低附着板中形成，并通过直接声滴喷射在每个化合物的九个半对数间隔浓度下进行给药。在 4 天的孵育后，用 ATP 终点法对 150 种具有已知 DILI 倾向的药物进行球体活力定量，这些药物的浓度间隔为半对数。我们为每个细胞系定义了安全性裕度，定义为为每个化合物生成的 IC 值与导致最佳分类准确性的最大血浆浓度 C 的比值。安全性裕度可用于估计早期药物发现中 C 尚未确定的化合物的最大安全 C。两种细胞系在预测 DILI 方面具有相似的准确性，其中 HepG2 球体更敏感。HepG2 球体的敏感性为 58%，特异性为 83%，而 HepaRG 球体的敏感性为 47%，特异性为 86%。150 种化合物中的 99 种用于比较 HepG2 和 HepaRG 球体的相对敏感性。HepaRG 球体对 7 种化合物更敏感，而 HepG2 球体对 34 种化合物更敏感。总之，在一组多样化的药物中，与 HepaRG 球体相比，HepG2 球体对 DILI 的预测性更高。
