title: Mechanism(s) regulating inhibition of thymidylate synthase and growth by gamma-L-glutaminyl-4-hydroxy-3-iodobenzene, a novel melanin precursor, in melanogenic melanoma cells.
abstract: A proposed mechanism for the melanoma specific activity of phenolic amines is based upon the ability of the enzyme tyrosinase to oxidize these prodrugs to toxic intermediates. In this study, we synthesized an iodinated analog of gamma-L-glutaminyl-4-hydroxybenzene (GHB) with increased antimelanoma activity in both human and murine melanoma cell lines. GHB and gamma-L-glutaminyl-4-hydroxy-3-iodobenzene (I-GHB) were shown to be substrates for both mammalian and mushroom tyrosinase. Glutathione, a cellular antioxidant, inhibited tyrosinase mediated formation of gamma-L-glutaminyl-3,4-benzoquinone (GBQ) from GHB, inhibited melanin production, and blocked the inhibition of the enzyme thymidylate synthase by oxidized GHB. Buthionine sulfoximine (BSO) depletion of cellular glutathione enhanced the growth inhibitory activity and the inhibition of in situ thymidylate synthase by phenolic amines in melanoma cells. GHB and I-GHB were shown to be approximately 5- and 10-fold more cytotoxic, respectively, in highly metastatic B16-BL6 cells than in weakly metastatic B16-F1 cells with approximately equal tyrosinase activity. B16-BL6 cells had approximately 20-fold higher gamma-glutamyltranspeptidase (gamma-GTPase) activity than B16-F1 cells which suggested the possible involvement of this enzyme in the activation of the cytotoxicity of the phenolic amines. 4-Aminophenol, a product of gamma-GTPase reaction with GHB, was a substrate for tyrosinase and a potent inhibitor of in situ thymidylate synthase activity in melanogenic cells. In pigmented melanoma cells containing the enzyme tyrosinase, the quinone mediated mechanism of phenolic amine cytotoxicity may be uniquely important and the cellular antioxidant glutathione essential in the detoxification of these quinone-generated intermediates.
authors.affiliation: Department of Radiation Oncology, University of Pittsburgh, PA.
authors.forename: J A
authors.initials: JA
authors.lastname: Prezioso
authors.forename: K M
authors.initials: KM
authors.lastname: Damodaran
authors.forename: N
authors.initials: N
authors.lastname: Wang
authors.forename: W D
authors.initials: WD
authors.lastname: Bloomer
mesh_terms: D000818:Animals; D001711:Biotransformation; D019328:Buthionine Sulfoximine; D002455:Cell Division / Q000187:drug effects; D020558:GTP Phosphohydrolases / Q000032:analysis; D005973:Glutamine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D005978:Glutathione / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D008543:Melanins / Q000096:biosynthesis*; D008545:Melanoma / Q000201:enzymology*; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors / Q000378:metabolism; D010084:Oxidation-Reduction; D010636:Phenols / Q000138:chemical synthesis / Q000494:pharmacology*; D011498:Protein Precursors / Q000378:metabolism*; D013778:Tetrazolium Salts; D013844:Thiazoles; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism
chemical_list: D008543:Melanins; D010636:Phenols; D011498:Protein Precursors; D013778:Tetrazolium Salts; D013844:Thiazoles; D005973:Glutamine; C079662:gamma-glutaminyl-4-hydroxy-3-iodobenzene; D008717:Methionine Sulfoximine; C001298:gamma-L-glutaminyl-4-hydroxybenzene; D019328:Buthionine Sulfoximine; D014442:Monophenol Monooxygenase; D013940:Thymidylate Synthase; D020558:GTP Phosphohydrolases; C022616:thiazolyl blue; D005978:Glutathione