Determination of CYP3A4 Inducing Properties of Compounds Using a Laboratory-Developed Cell-Based Assay.
A cell-based assay to measure cytochrome P450 3A4 (CYP3A4) induction was developed to screen for potential CYP3A4 inducers. This 96-well format assay utilizes HepG2 cells transfected with a gene construct of CYP3A4 proximal promoter linked to green fluorescence protein (GFP) gene, and the expression of the GFP is then measured quantitatively. Bergamottin at 5 to 25 µmol/L produced low induction relative to the positive control. Both curcumin and lycopene were not found to affect the expression of GFP, suggesting no induction properties toward CYP3A4. Interestingly, resveratrol produced significant induction from 25 µmol/L onward, which was similar to omeprazole and may warrant further studies. In conclusion, the present study demonstrated that this cell-based assay can be used as a tool to evaluate the potential CYP3A4 induction properties of compounds. However, molecular docking data have not provided satisfactory pointers to differentiate between CYP3A4 inducers from noninducers or from inhibitors, more comprehensive molecular screening may be indicated.
Malaysian Institute of Pharmaceuticals and Nutraceuticals, Ministry of Science, Technology and Innovation, Pulau Pinang, Malaysia.
Tiong Chai
TC
Seah
Malaysian Institute of Pharmaceuticals and Nutraceuticals, Ministry of Science, Technology and Innovation, Pulau Pinang, Malaysia.
Yea Lu
YL
Tay
Malaysian Institute of Pharmaceuticals and Nutraceuticals, Ministry of Science, Technology and Innovation, Pulau Pinang, Malaysia.
Heng Kean
HK
Tan
School of Fundamental Science, Universiti Malaysia Terengganu, Kuala Terengganu, Terengganu, Malaysia.
Tengku Sifzizul Tengku
TS
Muhammad
School of Pharmaceutical Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia.
Habibah Abdul
HA
Wahab
Malaysian Institute of Pharmaceuticals and Nutraceuticals, Ministry of Science, Technology and Innovation, Pulau Pinang, Malaysia Advanced Medical and Dental Institute, Universiti Sains Malaysia, Pulau Pinang, Malaysia tanml@usm.my.
Mei Lan
ML
Tan
D001681:Biological Assay; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis* / Q000235:genetics; D065693:Cytochrome P-450 Enzyme Inducers / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D007753:Laboratories; D062105:Molecular Docking Simulation; D011401:Promoter Regions, Genetic
D065693:Cytochrome P-450 Enzyme Inducers; D049452:Green Fluorescent Proteins; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human
CYP3A4 induction; cell-based assay; drug–drug interaction; drug–herb interaction