Target identification of drug induced mitochondrial toxicity using immunocapture based OXPHOS activity assays.
Mitochondrial dysfunction has been shown to be a pharmacotoxicological response to a variety of currently-marketed drugs. In order to reduce attrition due to mitochondrial toxicity, high throughput-applicable screens are needed for early stage drug discovery. We describe, here, a set of immunocapture based assays to identify compounds that directly inhibit four of the oxidative phosphorylation (OXPHOS) complexes: I, II, IV, and V. Intra- and inter-assay variation were determined and specificity tested by using classical mitochondrial inhibitors. Twenty drugs, some with known mitochondrial toxicity and others with no known mitochondrial liability, were studied. Direct inhibition of one or more of the OXPHOS complexes was identified for many of the drugs. Novel information was obtained for several drugs including ones with previously unknown effects on oxidative phosphorylation. A major advantage of the immunocapture approach is that it can be used throughout drug screening from early compound evaluation to clinical trials.
Nadanaciva
Sashi
S
MitoSciences Inc, 1850 Millrace Drive, Eugene, OR 97403, USA. snadanaciva@mitosciences.com
Bernal
Autumn
A
Aggeler
Robert
R
Capaldi
Roderick
R
Will
Yvonne
Y
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D004353:Drug Evaluation, Preclinical / Q000379:methods; D042967:Electron Transport Complex I / Q000037:antagonists & inhibitors / Q000378:metabolism; D042963:Electron Transport Complex II / Q000037:antagonists & inhibitors / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000633:toxicity; D007120:Immunochemistry; D066298:In Vitro Techniques; D008929:Mitochondria, Heart / Q000187:drug effects*; D009840:Oligomycins / Q000633:toxicity; D010085:Oxidative Phosphorylation / Q000187:drug effects*; D011190:Potassium Cyanide / Q000633:toxicity; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism; D012402:Rotenone / Q000633:toxicity; D013384:Succinate Cytochrome c Oxidoreductase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013804:Thenoyltrifluoroacetone / Q000633:toxicity; D014475:Uncoupling Agents / Q000633:toxicity*
D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D009840:Oligomycins; D014475:Uncoupling Agents; D012402:Rotenone; D013804:Thenoyltrifluoroacetone; D013384:Succinate Cytochrome c Oxidoreductase; D042963:Electron Transport Complex II; D006180:Proton-Translocating ATPases; D042967:Electron Transport Complex I; D011190:Potassium Cyanide