title: Metabolism of capsaicin by cytochrome P450 produces novel dehydrogenated metabolites and decreases cytotoxicity to lung and liver cells.
abstract: Capsaicin is a common dietary constituent and a popular homeopathic treatment for chronic pain. Exposure to capsaicin has been shown to cause various dose-dependent acute physiological responses including the sensation of burning and pain, respiratory depression, and death. In this study, the P450-dependent metabolism of capsaicin by recombinant P450 enzymes and hepatic and lung microsomes from various species, including humans, was determined. A combination of LC/MS, LC/MS/MS, and LC/NMR was used to identify several metabolites of capsaicin that were generated by aromatic (M5 and M7) and alkyl hydroxylation (M2 and M3), O-demethylation (M6), N- (M9) and alkyl dehydrogenation (M1 and M4), and an additional ring oxygenation of M9 (M8). Dehydrogenation of capsaicin was a novel metabolic pathway and produced unique macrocyclic, diene, and imide metabolites. Metabolism of capsaicin by microsomes was inhibited by the nonselective P450 inhibitor 1-aminobenzotriazole (1-ABT). Metabolism was catalyzed by CYP1A1, 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4. Addition of GSH (2 mM) to microsomal incubations stimulated the metabolism of capsaicin and trapped several reactive electrophilic intermediates as their GSH adducts. These results suggested that reactive intermediates, which inactivated certain P450 enzymes, were produced during catalytic turnover. Comparison of the rate and types of metabolites produced from capsaicin and its analogue, nonivamide, demonstrated similar pathways in the P450-dependent metabolism of these two capsaicinoids. However, production of the dehydrogenated (M4), macrocyclic (M1), and omega-1-hydroxylated (M3) metabolites was not observed for nonivamide. These differences may be reflective of the mechanism of formation of these metabolites of capsaicin. The role of metabolism in the cytotoxicity of capsaicin and nonivamide was also assessed in cultured lung and liver cells. Lung cells were markedly more sensitive to cytotoxicity by capsaicin and nonivamide. Cytotoxicity was enhanced 5 and 40% for both compounds by 1-ABT in BEAS-2B and HepG2, respectively. These data suggested that metabolism of capsaicinoids by P450 in cells represented a detoxification mechanism (in contrast to bioactivation).
authors.affiliation: Department of Pharmacology and Toxicology, Center for Human Toxicology, University of Utah, Salt Lake City, Utah 84112, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Reilly
authors.forename: William J
authors.initials: WJ
authors.lastname: Ehlhardt
authors.forename: David A
authors.initials: DA
authors.lastname: Jackson
authors.forename: Palaniappan
authors.initials: P
authors.lastname: Kulanthaivel
authors.forename: Abdul E
authors.initials: AE
authors.lastname: Mutlib
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Espina
authors.forename: David E
authors.initials: DE
authors.lastname: Moody
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: Crouch
authors.forename: Garold S
authors.initials: GS
authors.lastname: Yost
mesh_terms: D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D002211:Capsaicin / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity*; D002478:Cells, Cultured; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000737:chemistry / Q000494:pharmacology; D006041:Goats; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D006865:Hydrogenation; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011817:Rabbits; D051381:Rats; D014230:Triazoles / Q000494:pharmacology
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D014230:Triazoles; C033020:1-aminobenzotriazole; D003577:Cytochrome P-450 Enzyme System; D005978:Glutathione; D002211:Capsaicin