Measuring Drug Metabolism Efficiency of P450 Enzyme
Abstract
Cytochrome P450 enzyme is responsible for the majority of drug metabolism in the human body, generally increasing the drugs’ solubility for secretion. Drug companies have routinely tested how fast drugs are metabolized and the metabolites they produce, but the low efficiency of drug metabolism makes this measurement a challenging task. The purpose of this project was to increase metabolism efficiency by changing the reaction conditions. The drug metabolism of cytochrome P450 enzyme was measured using commonly prescribed medications. The P450 enzyme uses NADPH to reduce the heme iron- O2 complex making it highly reactive. This highly reactive center is responsible for oxidizing the drug; however, it can also produce superoxide or hydrogen peroxide from the bound oxygen, which decreases the enzyme’s efficiency. The goal of this project was to create a reducing environment for the enzyme to minimize side reactions. In this approach, NADPH was saved and redirected to only perform drug metabolism. NADPH was supplied through a regenerative system to the P450 liver microsomes by glucose 6-phosphate dehydrogenase enzyme. This enzyme reduces NADP+ to NADPH by oxidizing glucose 6-phosphate. The reducing environment could also neutralize the damages caused by hydrogen peroxide to the P450 enzyme. The substrate 2,6-di-tert-butyl-4-methylphenol (BHT) was used as a standard, while testing the efficiency of different drugs. The metabolites were detected and quantified using gas chromatography- mass spectrometry (GC-MS) and served as evidence that metabolism occurred.
Faculty Sponsors
Dr. Reza Razeghifard
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-3-2024 12:30 PM
End Date
4-4-2024 1:30 PM
Measuring Drug Metabolism Efficiency of P450 Enzyme
Alvin Sherman Library
Cytochrome P450 enzyme is responsible for the majority of drug metabolism in the human body, generally increasing the drugs’ solubility for secretion. Drug companies have routinely tested how fast drugs are metabolized and the metabolites they produce, but the low efficiency of drug metabolism makes this measurement a challenging task. The purpose of this project was to increase metabolism efficiency by changing the reaction conditions. The drug metabolism of cytochrome P450 enzyme was measured using commonly prescribed medications. The P450 enzyme uses NADPH to reduce the heme iron- O2 complex making it highly reactive. This highly reactive center is responsible for oxidizing the drug; however, it can also produce superoxide or hydrogen peroxide from the bound oxygen, which decreases the enzyme’s efficiency. The goal of this project was to create a reducing environment for the enzyme to minimize side reactions. In this approach, NADPH was saved and redirected to only perform drug metabolism. NADPH was supplied through a regenerative system to the P450 liver microsomes by glucose 6-phosphate dehydrogenase enzyme. This enzyme reduces NADP+ to NADPH by oxidizing glucose 6-phosphate. The reducing environment could also neutralize the damages caused by hydrogen peroxide to the P450 enzyme. The substrate 2,6-di-tert-butyl-4-methylphenol (BHT) was used as a standard, while testing the efficiency of different drugs. The metabolites were detected and quantified using gas chromatography- mass spectrometry (GC-MS) and served as evidence that metabolism occurred.
