title: Inactivation of anthracyclines by serum heme proteins.
abstract: We have previously shown that the anticancer agent doxorubicin undergoes oxidation and inactivation when exposed to myeloperoxidase-containing human leukemia HL-60 cells, or to isolated myeloperoxidase, in the presence of hydrogen peroxide and nitrite. In the current study we report that commercial fetal bovine serum (FBS) alone oxidizes doxorubicin in the presence of hydrogen peroxide and that nitrite accelerates this oxidation. The efficacy of inactivation was dependent on the concentration of serum present; no reaction was observed when hydrogen peroxide or serum was omitted. Peroxidase activity assays, based on oxidation of 3,3',5,5'-tetramethylbenzidine, confirmed the presence of a peroxidase in the sera from several suppliers. The peroxidative activity was contained in the >10000 MW fraction. We also found that hemoglobin, a heme protein likely to be present in commercial FBS, is capable of oxidizing doxorubicin in the presence of hydrogen peroxide and that nitrite further stimulates the reaction. In contrast to intact doxorubicin, the serum + hydrogen peroxide + nitrite treated drug appeared to be nontoxic for PC3 human prostate cancer cells. Together, this study shows that (pseudo)peroxidases present in sera catalyze oxidation of doxorubicin by hydrogen peroxide and that this diminishes the tumoricidal activity of the anthracycline, at least in in vitro settings. Finally, this study also points out that addition of H2O2 to media containing FBS will stimulate peroxidase-type of reactions, which may affect cytotoxic properties of studied compounds.
authors.affiliation: Department of Internal Medicine, University of Iowa, USA.
authors.forename: Brett A
authors.initials: BA
authors.lastname: Wagner
authors.forename: Lynn M
authors.initials: LM
authors.lastname: Teesch
authors.forename: Garry R
authors.initials: GR
authors.lastname: Buettner
authors.forename: Bradley E
authors.initials: BE
authors.lastname: Britigan
authors.forename: C Patrick
authors.initials: CP
authors.lastname: Burns
authors.forename: Krzysztof J
authors.initials: KJ
authors.lastname: Reszka
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology; D018943:Anthracyclines / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001560:Benzidines / Q000494:pharmacology; D001798:Blood Proteins / Q000737:chemistry* / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D004319:Doxylamine / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006420:Hemeproteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013058:Mass Spectrometry; D008706:Methemoglobin / Q000737:chemistry / Q000378:metabolism; D008713:Methimazole / Q000494:pharmacology; D015394:Molecular Structure; D010084:Oxidation-Reduction / Q000187:drug effects; D010544:Peroxidases / Q000737:chemistry / Q000378:metabolism; D010795:Phthalic Acids / Q000737:chemistry / Q000378:metabolism; D012459:Salicylates / Q000737:chemistry / Q000378:metabolism; D012977:Sodium Nitrite / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
chemical_list: C483040:3-methoxyphthalic acid; D000814:Aniline Compounds; D018943:Anthracyclines; D000903:Antibiotics, Antineoplastic; D001560:Benzidines; D001798:Blood Proteins; D006420:Hemeproteins; D010795:Phthalic Acids; D012459:Salicylates; C021758:3,3',5,5'-tetramethylbenzidine; C523953:3-methoxysalicylic acid; C034343:4-aminobenzhydrazide; D008713:Methimazole; D004317:Doxorubicin; D008706:Methemoglobin; D004319:Doxylamine; D006861:Hydrogen Peroxide; D010544:Peroxidases; D012977:Sodium Nitrite