Pharmakoökonomie: Open Access

Biotransformation monitoring, protein-ligand interactions and In-cell NMR spectroscopy in natural products research

Abstract

I.P. Gerothanassis

A critical overview of the following developments of NMR spectroscopy will be summarized:
(a) rapid ‘in situ’ monitoring of enzymatic reaction products [1], and enriching the biological space of natural products, through real time biotransformation monitoring in the
NMR tube. Investigation of interactions with the intrinsically disordered protein α-synuclein which is abundant in the human brain and a relevant target for neurodegenerative
diseases [2].
(b) The combined use of saturation transfer difference (STD), Tr-NOESY and INPHARMA (Interligand Noes for PHArmacophore MApping) NMR techniques for mapping
interactions, specific binding sites and structure elucidation of lipids with non-labelled serum albumin and the anti-apoptotic protein Bcl-2 [3,4].
(c) Application of in-cell NMR analytical methodology in the monitoring of the interaction of ligands with Bcl-2 inside living human cancer cells without requiring prior isotopic
labeling of the target protein. STD and Tr-NOESY NMR were employed to evaluate the direct binding of the ligand to the nonlabelled Bcl-2 protein intracellularly [5], which
was further validated in vitro. This approach has proved a very promising strategy for the real-time screening of the interaction profiling of drugs with their therapeutic targets
in their native cellular environment in living eukaryotic cells, paving the way to the new field of intracellular rational drug design.