Microbial production of phase I and phase II metabolites of midazolam

Abstract

1. Midazolam, a potent benzodiazepine derivative and a typical substrate of CYP3A4/3A5, is essentially metabolized in human into 1′-hydroxymidazolam, then eliminated as the corresponding phase II metabolite, the 1′-O-β-d-glucuronide derivative.

2. A high yield alternative to the current multistep synthesis of 1′-hydroxymidazolam is described, using a biotransformation of midazolam by a fungal microorganism, Beauveria bassiana.

3. The corresponding phase II metabolite, 1′-hydroxymidazolam-1′-O-β-d-glucuronide, has been then prepared by chemical synthesis (3 steps, 20% yield), or by microbial glucuronidation (one step, 20% yield) using a Streptomyces sp. strain.

4. The use of the same Streptomyces strain allows a direct and expeditive synthesis of the same glucuronide conjugate from midazolam itself in an advantageous 17% yield.

Keywords::

• 1′-hydroxymidazolam
• hydroxylation
• CYP3A4
• CYP3A5
• Beauveria bassiana
• Streptomyces sp.
• 1′-hydroxymidazolam-1′-O-β-d-glucuronide
• glucuronidation