High throughput screening campaign
Please go to Researchers involved for a list of researchers who contributed to this work
2023-10-25
This series was initiated by a high-throughput screen (HTS) on an Enamine 6K covalent library.The assay used was a DENV2 Rdrp picogreen assay, looking at single-point % inhibition. The compounds that fulfilled the criteria: > Average inhibition HTS + 3×SD, looking only at values between 0-100%, were included in the selection. The cut-off using this calculation was 60% inhibition, generating 123 compounds for selection. These compounds were grouped into 62 clusters based on similarity and one compound was selected from each cluster; these were submitted for DENV2 Rdrp IC50 in the picogreen assay.
The compounds represented 4 different covalent warheads outlined below. Warhead one was present in 33 of the tested compounds, warhead two in 8, warhead three in 5, and warhead four in 16.
Of the 62 compounds tested for DENV2 Rdrp IC50, 22 compounds showed an IC50 below 100 μM; 16 compounds with IC50 below 30 μM. The most hits were with warhead 1 (10 compounds), while warhead 4 only produced 3 hits. As the prevalence seems to be for warhead 1 and 2 these will be the focus of further development. Notable for all hits are that they are all more fragment-like and an expansion should probably focus on expanding the compounds to try and probe the size-limitation of binding pocket.
2023-11-01
Initial expansion of the two top warheads (1 and 2) was done by analyzing the HTS results in DataWarrior and looking for general trends in the data to guide expansion of the preliminary hits. The results are summarized below:
These efforts were terminated due to several compounds being false positives, please see Assay Interference