Preclinical Evaluations of Drug Candidates Using a Cell Viability Assay and Sensorimotor Evaluation Study
Mentor 1
Alexander Arnold
Start Date
28-4-2023 12:00 AM
Description
Preclinical evaluation following the synthesis of potential drugs candidates is an important part of drug discovery. The purpose of preclinical evaluation is to determine safety and adverse effects of new compounds. This evaluation includes two components: a cytotoxicity assay and sensorimotor study call the rotarod test. Cytotoxicity assays are used to determine the concentration of a new compound which induces 50% of cell death. For this purpose, human kidney cells plated in 384 well plates are combined with serial diluted compounds dissolved in dimethyl sulfoxide. The treated cell plate is then incubated for 18 hours before the addition of CellTiter Glo, a luminescence-based viability assay. Given the amount of ATP remaining in the living cells, a fluorescent glow can be quantified. The data was analyzed by non-linear regression to calculate LD50 values, which represent the compound concentration that produces 50% cell death. The rotarod tests is a sensorimotor test to evaluate a given compound’s effect on the central nervus system. Mice are trained to balance on a rotating rod for three minutes without falling. For the evaluation, compounds are administered orally by gavage and then placed on a rotating rod at 10, 30, and 60 minutes after administration for three minute intervals. Compounds that inhibit sensorimotor coordination will prevent animal to balance for three minutes. Data was analyzed by ANOVA repeated measures.
Preclinical Evaluations of Drug Candidates Using a Cell Viability Assay and Sensorimotor Evaluation Study
Preclinical evaluation following the synthesis of potential drugs candidates is an important part of drug discovery. The purpose of preclinical evaluation is to determine safety and adverse effects of new compounds. This evaluation includes two components: a cytotoxicity assay and sensorimotor study call the rotarod test. Cytotoxicity assays are used to determine the concentration of a new compound which induces 50% of cell death. For this purpose, human kidney cells plated in 384 well plates are combined with serial diluted compounds dissolved in dimethyl sulfoxide. The treated cell plate is then incubated for 18 hours before the addition of CellTiter Glo, a luminescence-based viability assay. Given the amount of ATP remaining in the living cells, a fluorescent glow can be quantified. The data was analyzed by non-linear regression to calculate LD50 values, which represent the compound concentration that produces 50% cell death. The rotarod tests is a sensorimotor test to evaluate a given compound’s effect on the central nervus system. Mice are trained to balance on a rotating rod for three minutes without falling. For the evaluation, compounds are administered orally by gavage and then placed on a rotating rod at 10, 30, and 60 minutes after administration for three minute intervals. Compounds that inhibit sensorimotor coordination will prevent animal to balance for three minutes. Data was analyzed by ANOVA repeated measures.
