Oxidative coupling of mithramycin and hydroquinone catalyzed by copper oxidases and benzoquinone. Implications for the mechanism of action of aureolic acid antibiotics.
The copper oxidases human ceruloplasmin and Polyporous anceps laccase catalyze the oxidative coupling of mithramycin (1) and its aglycone chromomycinone (2) with p-hydroquinone to form new mithramycin-hydroquinone (3) and chromomycinone-hydroquinone adducts (4), respectively. Similar adducts could be formed by the nonenzymatic mimic of this reaction using benzoquinone and these aureolic acids in buffer solutions. FABMS of 3 indicated that the hydroquinone moiety was attached to the aureolic acid aglycone. Acid hydrolysis of 3 yielded a compound with the same chromatographic and spectroscopic characteristics as 4. Structure elucidation of 4 by NMR and MS revealed that the hydroquinone was attached to the C-5 position of the aglycone. NMR evidence indicated that 4 consisted of a mixture of ortho-substituted biphenyl rotamers. The mechanism of the copper oxidase catalyzed adduct formation reaction is presumed to involve radical formation through hydrogen removal at the 8-phenolic position, radical isomerization, and coupling with semiquinone radical also formed during enzymatic and nonenzymatic incubations. Identification of the covalent-hydroquinone adduct provides evidence that aureolic acid antibiotics can be metabolically converted to reactive radical intermediates, and it establishes the C-5 position of aureolic acid as an enzymatically reactive site. Unlike mithramycin, the mithramycin-hydroquinone adducts was inactive in the in vivo P388 leukemic antitumor test system.
Anyanwutaku
I O
IO
Division of Medicinal and Natural Products Chemistry, University of Iowa, College of Pharmacy, Iowa City 52242.
Petroski
R J
RJ
Rosazza
J P
JP
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000138:chemical synthesis* / Q000737:chemistry / Q000627:therapeutic use; D016227:Benzoquinones / Q000737:chemistry*; D002384:Catalysis; D002570:Ceruloplasmin / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical; D006801:Humans; D006873:Hydroquinones / Q000737:chemistry*; D042845:Laccase; D007941:Leukemia P388 / Q000188:drug therapy; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D015394:Molecular Structure; D010088:Oxidoreductases / Q000378:metabolism*; D008926:Plicamycin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000627:therapeutic use; D011124:Polyporaceae / Q000201:enzymology*; D016339:Spectrometry, Mass, Fast Atom Bombardment
D000903:Antibiotics, Antineoplastic; D016227:Benzoquinones; D006873:Hydroquinones; C004532:quinone; D010088:Oxidoreductases; D042845:Laccase; C024748:copper oxidase; D002570:Ceruloplasmin; D008926:Plicamycin; C031927:hydroquinone