An integrated array chip for high-throughput screening species differences in metabolism.

Species differences in metabolism may produce failure prediction of drug efficacy/toxicity in humans. Integration of metabolic competence and cellular effects assay in vitro can provide insight into the species differences in metabolism, however, require a co-culture platform with features of high-throughput, operationally simple, low sample consumption, and independent layouts for potential usage in industrial test settings. Herein, we developed an integrated array chip (IAC) to evaluate the species differences in metabolism through metabolism-induced anticancer bioactivity as a case. The IAC consisted of two functional parts of a micropillar chip for immobilization of liver microsomes and a microwell chip for 3D tumor cell culture. Firstly, optimized parameters of the micropillar chip for microsomal encapsulation were obtained by cross-shaped protrusion and 2.5 μL volume of 3D agarose spot. Next, we examined factors influencing metabolism-induced anticancer bioactivity. Feasibility of the IAC was validated by four model prodrugs using image-based bioactivity detection and mass spectrometry (MS)-based metabolites analysis. Finally, species-specific IAC was used for selection of animal species that best resembles metabolism-induced drug response to humans at throughputs. Overall, the IAC provides a promising co-culture platform for identifying species differences in metabolism and selection of animal models to accelerate drug discovery.