Identification of potential TMPRSS2 inhibitors via high-throughput screening based on oriented immobilized cell membrane chromatography technology.

The transmembrane protease serine 2 (TMPRSS2) plays a crucial role in the cellular entry of coronaviruses, making the search for its inhibitors pertinent for developing novel antiviral drugs. Cell membrane chromatography (CMC) is a novel methodology that immobilizes membrane receptors on silica gel, utilizing chromatographic techniques to discover new drugs. To enhance the accuracy of this method, this study employed styrene-maleic acid (SMA) copolymers for protein extraction His-tag for protein immobilization, which would minimize alterations to the biological structure of TMPRSS2. Methodological validation demonstrated that this model offers improved reproducibility and longer column lifespan compared to traditional CMC columns, alongside superior screening performance. Utilizing this model, a screening campaign was conducted against a commercial small molecule library containing 3010 compounds. Preliminary activity validation revealed that the screened famotidine and TS0665 effectively inhibited pseudovirus infection of cells. These findings provide an experimental foundation for the subsequent development of antiviral therapeutics.