Laccase-mediated synthesis of 2-methoxy-3-methyl-5-(alkylamino)- and 3-methyl-2,5-bis(alkylamino)-[1,4]-benzoquinones

The synthesis of 5-alkylamino- and 2,5-bis(alkylamino)-[1,4]-benzoquinones, showing structural similarity to natural mitomycins, was performed through coupling of 2-methoxy-3-methylhydroquinone with primary amines such as n-octylamine, geranylamine and cyclooctylamine using laccases from Myceliophthora thermophila (MtL) and Pycnoporus cinnabarinus SBUG-M 1044 (PcL). Product spectra of laccase reactions differ due to reaction systems pH values (pH 7.0 for MtL and pH 5.0 for PcL) applied to assure enzymes optimal catalytic efficiency. The MtL- and PcL-mediated formation of monoaminated products was achieved at equimolar reactant concentrations with amine coupling at the meta-position to benzoquinones methyl group. Increased formation of diaminated products occurred in PcL-mediated reactions and generally when the amine was supplied in excess. Diamination entailed elimination of the benzoquinone methoxy group (amination in para-position to the first amine substituent). Six products were synthesised and characterised by NMR and HR-MS analysis. The laccase-mediated amine coupling to 2-methoxy-3-methylhydroquinone confers two of the essential pharmaceutical active motifs from mitomycins: (i) a stable 1,4-benzoquinoic parent structure and (ii) a biological active alkylation function (NH).

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► The laccase-mediated CN coupling reaction between 2-methoxy-3-methylhydroquinone and n-octylamine, geranylamine and cyclooctylamine has been investigated.
► Laccases from Myceliophthora thermophila (MtL) and Pycnoporus cinnabarinus (PcL) were used.
► Monoamination occurred at benzoquinones C-5 atom and preferentially at equal reactant concentrations and pH 7.0 using MtL.
► Diamination occurred with excess of amines and pH 5.0 using PcL, respectively, via elimination of benzoquinones methoxy group.
► Products revealed pharmaceutical active structure motifs with alikeness to natural occurring mitomycins.