title: Thiomers: Inhibition of cytochrome P450 activity.
abstract: The aim of the present study was to investigate the potential of different thiolated polymers (thiomers) on the catalytic activity of CYP450s on one hand and to explore new inhibitors for CYP activity on the other hand. Several thiolated polymers including poly(acrylic acid)-cysteine (PAA-cysteine), chitosan-thioglycolic acid (chitosan-TGA), and thiolated PEG-g-PEI copolymer along with brij 35, myrj 52 and the well-established CYPP450 inhibitor verapamil were screened for their CYP3A4 and CYP2A6 inhibitory activity, and their IC(50) values were determined. Both enzyme inhibition assays were performed in 96-well microtiter plates. 7-Benzyloxy-4-(trifluoromethyl)-coumarin (BFC) and 7-hydroxycoumarin (7-HC) were used as fluorescent substrates in order to determine CYP3A4 and CYP2A6 catalytic activity, respectively. All investigated compounds inhibited CYP3A4 as well as CYP2A6 activity. All tested (thiolated) polymers were found to be more potent inhibitors of CYP3A4 than of CYP2A6 catalytic activity. Apart from verapamil that is a known CYP3A4 inhibitor, brij 35 and myrj 52 were explored as potent inhibitors of CYP3A4 and CYP2A6 catalytic activity. Among the tested polymers, the rank order for CYP3A4 inhibition was PAA-cysteine (100 kDa)>brij 35>thiolated PEG-g-PEI copolymer (16 kDa)>myrj 52>PAA (100 kDa)>PAA-cysteine (450 kDa)>verapamil>PAA (450 kDa)>chitosan-TGA (150 kDa)>chitosan (150 kDa). On the other hand, the rank order of CYP2A6 inhibition was brij 35>PAA-cysteine (100kDa)>chitosan-TGA (150 kDa)>PAA (100 kDa)>thiolated PEG-g-PEI copolymer (16 kDa)>PAA-cysteine (450 kDa)>chitosan (150 kDa)>verapamil>PAA (450 kDa)>myrj 52. Thus, this study suggests that (thiolated) polymers display a promising potential to inhibit cytochrome P450s activity and might turn out to be potentially valuable tools for improving the oral bioavailability of actively secreted compounds by avoiding intestinal metabolism.
authors.affiliation: Department of Pharmaceutical Technology, University of Innsbruck, Innrain, Innsbruck, Austria.
authors.forename: Javed
authors.initials: J
authors.lastname: Iqbal
authors.forename: Duangkamon
authors.initials: D
authors.lastname: Sakloetsakun
authors.forename: Andreas
authors.initials: A
authors.lastname: Bernkop-Schnürch
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000037:antagonists & inhibitors / Q000378:metabolism; D048271:Chitosan / Q000737:chemistry; D003374:Coumarins / Q000737:chemistry; D003545:Cysteine / Q000032:analysis / Q000737:chemistry / Q000494:pharmacology; D065637:Cytochrome P-450 CYP2A6; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D065692:Cytochrome P-450 CYP3A Inhibitors; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004337:Drug Carriers; D004339:Drug Compounding; D016503:Drug Delivery Systems; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D005079:Excipients; D007202:Indicators and Reagents / Q000737:chemistry; D011108:Polymers / Q000737:chemistry* / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000737:chemistry* / Q000494:pharmacology; D014700:Verapamil / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
chemical_list: D003374:Coumarins; D065692:Cytochrome P-450 CYP3A Inhibitors; D065607:Cytochrome P-450 Enzyme Inhibitors; D004337:Drug Carriers; D004791:Enzyme Inhibitors; D005079:Excipients; D007202:Indicators and Reagents; D011108:Polymers; D013438:Sulfhydryl Compounds; D048271:Chitosan; D003577:Cytochrome P-450 Enzyme System; D014700:Verapamil; D001189:Aryl Hydrocarbon Hydroxylases; C585710:CYP2A6 protein, human; D065637:Cytochrome P-450 CYP2A6; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D003545:Cysteine