The potential of 19F NMR spectroscopy for rapid screening of cell cultures for models of mammalian drug metabolism

Abstract

The use of microbial cultures as a complementary model for mammalian drug metabolism has been well established previously. Here is a preliminary investigation into the potential of 19F NMR spectroscopy as a rapid screening tool to quantify the biotransformations of fluorine-containing model drugs. Biotransformations of three model drugs in 48 taxonomically diverse organisms were measured by acquiring 19F NMR spectra at 376 MHz. The presence of fluorine in the molecules allowed rapid, simultaneous detection of over 20 biotransformation products without sample pretreatment, chromatography, mass spectrometric techniques or the use of radiolabelled substrates. The detection limit at 376 MHz using 5 mm NMR tubes was ca. 0.3 microg ml(-1) using a typical analysis time of 20 min per sample. With the recent advent of flow injection NMR technology, analysis time of 5 min could be achieved with less sample. This approach may be used to develop fast small-scale microbial screens for the biosynthesis of metabolite standards and production of novel drug analogues, whilst also having a role in reducing animal experiments needed to identify animal and human metabolites of fluorinated xenobiotics.

19F NMR applications in chemical biology

“…Since animal models are becoming less attractive, microbial models of mammalian metabolism are increasingly used. Corcoran et al [33] examined the microbial transformation products of model fluorinated pharmaceutical compounds by 19 F NMR. Of the 48 strains examined, four effectively degraded the fluorinated compounds and the number and nature of the catabolites could be readily assessed using this technique, for example, 5-fluoroindole was transformed completely by Streptomyces sp.…”

Microscale nuclear magnetic resonance: a tool for soft matter research

“…155 Other remarkable applications include plant metabolite profiling through NMR spectroscopy, 156 single insect NMRanalysing the chemical composition of defensive secretions 157 and exploiting the combination of gas chromatography-mass spectrometry and NMR spectroscopy for the characterisation of dandelion species. 158 …”

Stable isotope-resolved metabolomics and applications for drug development

“…Metabolism of 19 F-containing drugs has been extensively studied by Nicholson's group (Wade, Wilson et al 1990; Spraul, Hofmann et al 1993; Corcoran, Lindon et al 2001; Blackledge, Nicholson et al 2002; Blackledge, Nicholson et al 2003; Keun, Athersuch et al 2008). In these studies, the agent was introduced to an organism such as a rat, followed by the analysis of the drug and its metabolites by 19 F and 1 H NMR in urine and or blood plasma.…”