Figure 3.

Schematic overview of the screening campaign used on the Janssen POPS library containing 5676 compounds. The high‐throughput screen was performed on 384‐well plates with the described PR or HCI VeroE6‐eGFP assay. After the initial screening, 228 compounds showed activity against SARS‐CoV‐2 with EC₅₀ values <20 µM in either assay. These compounds were retested in triplicate in the same assay for confirmation, which yielded 52 active compounds. Finally, the eight most promising compounds were selected for further profiling, and the top six candidates were selected for additional testing using the Caco‐2 assay. EC₅₀, concentration of the compound that inhibited 50% of the infection; HCI, high content imaging; POPS, Phase One Passed Structures; PR, plate reader; VeroE6‐eGFP, VeroE6 African green monkey kidney epithelial cells expressing a stable enhanced green fluorescent protein.