Application of a high-content multiparameter cytotoxicity assay to prioritize compounds based on toxicity potential in humans.
Prioritization of compounds based on human hepatotoxicity potential is currently a key unmet need in drug discovery, as it can become a major problem for several lead compounds in later stages of the drug discovery pipeline. The authors report the validation and implementation of a high-content multiparametric cytotoxicity assay based on simultaneous measurement of 8 key cell health indicators associated with nuclear morphology, plasma membrane integrity, mitochondrial function, and cell proliferation. Compounds are prioritized by (a) computing an in vitro safety margin using the minimum cytotoxic concentration (IC(20)) across all 8 indicators and cell-based efficacy data and (b) using the minimal cytotoxic concentration alone to take into account concentration of drug in tissues. Feasibility data using selected compounds, including quinolone antibiotics, thiazolidinediones, and statins, suggest the viability of this approach. To increase overall throughput of compound prioritization, the authors have identified the higher throughput, plate reader-based CyQUANT assay that is similar to the high-content screening (HCS) assay in sensitivity of measuring inhibition of cell proliferation. It is expected that the phenotypic output from the multiparametric HCS assay in combination with other highly sensitive approaches, such as microarray-based expression analysis of toxic signatures, will contribute to a better understanding and predictivity of human hepatotoxicity potential.
Abraham
Vivek C
VC
Lead Discovery, Abbott Laboratories, Abbott Park, IL 60064-6217, USA. vivek.abraham@abbott.com
Towne
Danli L
DL
Waring
Jeffrey F
JF
Warrior
Usha
U
Burns
David J
DJ
D001681:Biological Assay; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D064420:Drug-Related Side Effects and Adverse Reactions / Q000145:classification; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D020128:Inhibitory Concentration 50; D008113:Liver Neoplasms / Q000473:pathology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D018675:Toxicity Tests / Q000379:methods*; D015262:Xenobiotics / Q000145:classification / Q000633:toxicity*
D015262:Xenobiotics