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NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode

NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode

Published on: June 4, 2021

Fluorine NMR-based screening on cell membrane extracts.

Marina Veronesi¹, Elisa Romeo, Chiara Lambruschini

¹Department of Drug Discovery & Development, Fondazione Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa (Italy).

|December 17, 2013

Summary

This summary is machine-generated.

We introduce n-fluorine atoms for biochemical screening (n-FABS), a novel NMR method to detect enzyme inhibitors in cell samples. This technique enables lead discovery for difficult targets, advancing drug development.

Keywords:

19F NMR spectroscopy drug discovery fragments functional screening inhibitors membrane proteins

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Area of Science:

Biochemistry
Chemical Biology
Drug Discovery

Background:

Assaying enzyme inhibitors in physiological conditions is crucial for lead discovery.
Limited biophysical techniques are suitable for intact cells, cell lysates, or membrane preparations.
Developing versatile assays for complex biological systems is a significant challenge.

Purpose of the Study:

To demonstrate the first application of n-fluorine atoms for biochemical screening (n-FABS) using (19)F NMR spectroscopy.
To detect high- and low-affinity inhibitors of a membrane enzyme in cell extracts.
To determine IC50 values for identified inhibitors.

Main Methods:

Utilized n-fluorine atoms for biochemical screening (n-FABS).
Employed homogeneous (19)F NMR spectroscopy.
Applied the assay to cell extracts and membrane preparations containing a target enzyme.

Main Results:

Successfully detected high- and low-affinity inhibitors of a membrane enzyme.
Determined IC50 values for the inhibitors.
Demonstrated the versatility of n-FABS for complex biological samples.

Conclusions:

n-FABS is a novel, homogeneous, and versatile assay for detecting enzyme inhibitors in cell extracts.
This method facilitates the discovery of novel binding fragments for challenging targets, including membrane-associated proteins.
The findings pave the way for broader applications of (19)F NMR in drug discovery for complex biological systems.