Application Notes

APPLICATION NOTE 2 ©2019 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. Introduction The inefficiency of the drug discovery process is a widely accepted problem by everyone in the pharmaceutical industry. Investment in pharmaceutical research and development continues to increase, while the number of new drugs approved by the US Food and Drug Administration remains low 1 . One area that is crucial to making drug discovery more efficient is the screening process. Current trends demand technologies that can characterize binding events involving more heterogeneous targets and compound libraries that are not larger, but rather made up of more diverse molecules from large and new chemical spaces. There is also a need for faster tools, as computer-aided iterative screening is becoming more popular as an alternative to high throughput screening. Dianthus is the screening tool that makes drug discovery more cost-effective — it detects and quantifies molecular interactions in solution, independent of the mass and size of the two interacting molecules, and does it at an unmatched speed with flexible throughput. To quantify molecular interactions, Dianthus uses Temperature Related Intensity Change (TRIC), a technology with 10 years of proven success in molecular interaction research 2 . TRIC measures the strength of the interaction by detecting a variation in the fluorescence signal of a fluorescently labeled or intrinsically fluorescent target as a result of a very rapid and precise IR- laser induced temperature change. The binding of a ligand in close proximity to the fluorophore or a ligand-induced conformational change in the target strongly affects the extent of the change in fluorescence between the unbound (no ligand) and bound (with ligand) state of the target. Changes in fluorescence upon activation of the IR laser are monitored to characterize the interaction and derive affinity constants 2 . The fragment-based lead discovery study presented here, utilizes a library with 2,490 fragments from a large chemical space in two steps: single-dose screening and affinity screening. The target molecule, G9a (also known as EHMT2), has been the focus of oncological drug discovery research over the last decade and Irregularities in its activity have been associated with a variety of oncogenic phenotypes, such as breast or lung cancer 3 . G9a is a histone methyltransferase (HMT) that acts in methylation of H3K9 (lysine 9 in histone 3). HMTs are enzymes involved in epigenetic gene regulation by methylation of histone proteins in either Lysine or Arginine residues 4 . Finding inhibitors with different modes

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