Targeted Mass Spectrometry Approaches (RFMS and AEMS)

Targeted mass spectrometry based approaches such as the RapidFire^® High Throughput MS (RFMS) system and the Acoustic Ejection MS (AEMS) system are important platforms for assessing the biophysics of molecules and play a role in target-based drug discovery. Both mass spectrometry platforms provide rapid, high-quality data with the flexibility to analyse a range of different molecules.

The RapidFire^® High-throughput MS System (RFMS) provides fast and efficient analyses of functional biochemical assays and samples in biological matrices. The devices are running with 384-well plates. Typical throughput/day are approximately 6,000 samples/device and are part of Evotec’s Hit Identification platform.​

The readout is mass-based, and our MS-based devices can even differentiate between mass changes of 1 Da or less, if fragmentation patterns change sufficiently. The analysis of substrate/product analytes is currently possible for small molecules, metabolites, and peptides. Workflows for larger molecules like RNA or proteins have been established. ​

Acoustic Ejection Mass Spectrometry (AEMS) system delivers speed, scale and unyielding data quality to high-throughput screening laboratories working with complex biology. This technology was announced in 2019 and Sciex' product was launched in 2020. 2.5 nl droplets of sample solutions are directly ejected by the Echo device into the MS device boosting the throughput to more than 30,000 samples/day. The Sciex device is accepting 1,536- as well as 384-well plates.

The following list of case studies, published with collaborating partners, demonstrate the application of RFMS technology:

• Structural basis of SARM1 activation, substrate recognition, and inhibition by small molecules
• Pharmacological SARM1 inhibition protects axon structure and function in paclitaxel-induced peripheral neuropathy
• Discovery of novel UDP-N-acetylglucosamine acyltransferase (LpxA) inhibitors with activity against Pseudomonas aeruginosa
• Small-molecule inhibition of METTL3 as a strategy against myeloid leukaemia
• Development of a series of aryl pyrimidine kynurenine monooxygenase inhibitors as potential therapeutic agents for the treatment of Huntington’s Disease
• Tetrahydroindazoles as interleukin-2 inducible T-cell kinase inhibitors. Part II. Second-generation analogues with enhanced potency, selectivity, and pharmacodynamic modulation in vivo
• Property- and structure-guided discovery of a tetrahydroindazole series of interleukin-2 inducible T-cell kinase inhibitors
• Development of LC/MS/MS, high-throughput enzymatic and cellular assays for the characterization of compounds that inhibit kynurenine monooxygenase (KMO)
• A label-free LC/MS/MS-based enzymatic activity assay for the detection of genuine caspase inhibitors and SAR development