Untargeted drug screening via high-throughput metabolomics
2Computer Science, Technion - Israel Institute of Technology, Israel
Modern drug discovery typically involves target identification, validation, optimization, and preclinical development. In the last thirty years, high-throughput screening (HTS) has been widely used to eliminate hours of painstaking testing by scientists, improving the quality and accuracy of data. Only nuclear magnetic resonance (NMR) and mass spectrometry (MS) methods are suitable for untargeted metabolic in drug discovery. However, common LC-MS technics are not suitable for high throughput analysis because each sample requires 20-40 minutes.
We established a system for automated screening of drugs targeting metabolic enzymes that consist of the following steps:
1. Automated cell seeding in 96-well plates
2. Drug treatment in a series of dosages
3. Metabolomic extraction
4. Cell number estimation
5. Rapid untargeted metabolomics.
We established a rapid method for untargeted metabolomics with mass-spectrometry detection. We reduced the cycle time with an ultra-fast gradient to 2 minutes. A 2-minute LC analysis with an informative MS time of 1 minute allowed us to combine 2 columns with a cycle-shift time of 1 minute with one MS instrument continuously analyzing data from 2 LC columns working in parallel.
We assembled a list of 110 chemical inhibitors of metabolic enzymes, including FDA-approved drugs (40%), drugs in various phases of clinical trials (25%), and tool compounds (35%). We applied the developed method to explore the time- and dosage-dependent metabolic response to treatment with the above drugs. The analysis output allowed us to detect affected pathways and estimate the drug efficacy for the drugs sheared the same target.