Lead Compounds Discovery Of Histone Acetylation "Reader" ENL YEATS Domain

In recent decades,there have been many mature drugs for traditional diseases in the market,so the focus of drug research and development is gradually shifting from the traditional field to the emerging science or therapeutic field which can open up new opportunities.At the same time,it benefits from the development and application of novel targets and fresh technologies,the research of drug that use new and unique mechanisms to treat certain diseases has attracted more and more attention.As a novel histone acetylation"reader",ENL YEATS domain has been reported to be closely related to the occurrence and development of many diseases in recent years,and is a potential drug target.However,the research progress of the lead compounds targeting the protein is relatively slow.Up to now,there is still no safe,reliable and effective drug targeting ENL YEATS domain protein.Therefore,the discovery of lead compounds targeting ENL YEATS domain is still a hot research topic that needs to be solved urgently.In this paper,we focused on ENL YEATS domain,a new target in epigenetic field,established a high-throughput screening platform based on ALPHAScreen technology,and screened small molecule inhibitors from the in-house compound library.After a series of repeated experiments,we selected two kinds of compounds with strong activity for characterization,and named them compound 1 and compound 3-6respectively.The IC₅₀ values of the two compounds were 7.3±1.7μM and 124.1±17.6 n M,respectively.Next,surface plasmon resonance（SPR）and protein thermal shift（PTS）,microscale thermophoresis（MST）and hydrogen deuterium exchange mass spectrometry（HDX-MS）were used to verify the binding ability of the two compounds.Then,combined with the biochemical activity verification results,compound 3-6 with better activity in vitro was selected for a series of cell level antitumor activity evaluation.The results showed that compound 3-6 could inhibit the proliferation and induce apoptosis of MV4-11 cell line,and significantly reduce the transcription and expression of its target oncogenes.In conclusion,in this paper,we constructed a high-throughput screening platform with great stability and high sensitivity,and found two kinds of lead compounds targeting ENL YEATS domain,which have the advantages of novel skeleton and good inhibition effect.This work not only provides a new chemical probe for the treatment of acute myelogenous leukemia and other YEATS family proteins related diseases,but also provides ideas and references for the subsequent modification and optimization of pharmacochemistry based on two types of skeletons.