Toxicological evaluation of thiol-reactive compounds identified using a la assay to detect reactive molecules by nuclear magnetic resonance.
We have recently reported on the development of a La assay to detect reactive molecules by nuclear magnetic resonance (ALARM NMR) to detect reactive false positive hits from high-throughput screening, in which we observed a surprisingly large number of compounds that can oxidize or form covalent adducts with protein thiols groups. In the vast majority of these cases, the covalent interactions are largely nonspecific (e.g., affect many protein targets) and therefore unsuitable for drug development. However, certain thiol-reactive species do appear to inhibit the target of interest in a specific manner. The question then arises as to the potential toxicology risks of developing a drug that can react with protein thiol groups. Here, we report on the evaluation of a large set of ALARM-reactive and -nonreactive compounds against a panel of additional proteins (aldehyde dehydrogenase, superoxide dismutase, and three cytochrome P450 enzymes). It was observed that ALARM-reactive compounds have significantly increased risks of interacting with one or more of these enzymes in vitro. Thus, ALARM NMR seems to be a sensitive tool to rapidly identify compounds with an enhanced risk of producing side effects in humans, including alcohol intolerance, the formation of reactive oxygen species, and drug-drug interactions. In conjunction with other toxicology assays, ALARM NMR should be a valuable tool for prioritizing compounds for lead optimization and animal testing.
Huth
Jeffrey R
JR
Global Pharmaceutical Research and Development, Abbott Laboratories, Abbott Park, IL 60064, USA.
Song
Danying
D
Mendoza
Renaldo R
RR
Black-Schaefer
Candice L
CL
Mack
Jamey C
JC
Dorwin
Sarah A
SA
Ladror
Uri S
US
Severin
Jean M
JM
Walter
Karl A
KA
Bartley
Diane M
DM
Hajduk
Philip J
PJ
D000444:Aldehyde Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D001324:Autoantigens / Q000737:chemistry*; D065607:Cytochrome P-450 Enzyme Inhibitors; D015195:Drug Design; D064420:Drug-Related Side Effects and Adverse Reactions; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D015394:Molecular Structure; D004364:Pharmaceutical Preparations / Q000032:analysis; D011485:Protein Binding; D012261:Ribonucleoproteins / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry* / Q000378:metabolism; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000378:metabolism; D000097585:SS-B Antigen
D000444:Aldehyde Dehydrogenase; D001324:Autoantigens; D065607:Cytochrome P-450 Enzyme Inhibitors; D004364:Pharmaceutical Preparations; D012261:Ribonucleoproteins; D013438:Sulfhydryl Compounds; D013482:Superoxide Dismutase; D000097585:SS-B Antigen