title: Extractive biotransformation for production of metabolites of poorly soluble compounds: synthesis of 32-hydroxy-rifalazil.
abstract: A novel reaction system was developed for the production of metabolites of poorly water-soluble parent compounds using mammalian liver microsomes. The system includes the selection and use of an appropriate hydrophobic polymeric resin as a reservoir for the hydrophobic parent compounds and its metabolites. The utility of the extractive biotransformation approach was shown for the production of a low-yielding, synthetically challenging 32-hydroxylated metabolite of the antibiotic rifalazil using mouse liver microsomes. To address the low solubility and reactivity of rifalazil in the predominantly aqueous microsomal catalytic system, a variety of strategies were tested for the enhanced delivery of hydrophobic substrates, including the addition of mild detergents, polyvinylpyrrolidone, glycerol, bovine serum albumin, and hydrophobic polymeric resins. The latter strategy was identified as the most suitable for the production of 32-hydroxy-rifalazil, resulting in up to 13-fold enhancement of the volumetric productivity compared with the standard aqueous system operating at the solubility limit of rifalazil. The production process was optimized for a wide range of reaction parameters; the most important for improving volumetric productivity included the type and amount of the polymeric resin, cofactor recycling system, concentrations of the biocatalyst and rifalazil, reaction temperature, and agitation rate. The optimized extractive biotransformation system was used to synthesize 32-hydroxy-rifalazil on a multimilligram scale.
authors.affiliation: AMRI, Department of Metabolism and Biotransformations, 21 Corporate Circle, P.O. Box 15098, Albany, NY 12212-5098, USA. dima.mozhaev@amriglobal.com
authors.forename: Vadim V
authors.initials: VV
authors.lastname: Mozhaev
authors.forename: Lyudmila V
authors.initials: LV
authors.lastname: Mozhaeva
authors.forename: Peter C
authors.initials: PC
authors.lastname: Michels
authors.forename: Yuri L
authors.initials: YL
authors.lastname: Khmelnitsky
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis* / Q000493:pharmacokinetics; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D005260:Female; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism; D012294:Rifamycins / Q000138:chemical synthesis* / Q000493:pharmacokinetics; D012995:Solubility
chemical_list: C533852:32-hydroxyrifalazil; D000900:Anti-Bacterial Agents; D012294:Rifamycins