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BODIPY-Tagged β-Lactams as Selective Quenched Activity-Based Probes to Target Human Neutrophil Elastase

Felix, R.; Carvalho, L. A. R.; Guedes, R.; Madureira, A. M.; Mallo-Abreu, A.; Goncalves, L.; Genilloud, O.; Fernandez-Godino, R.; Ramos, M. C.; Moreira, R.

2026-03-22 pharmacology and toxicology
10.64898/2026.03.19.712884 bioRxiv
Show abstract

Human Neutrophil Elastase (HNE) plays a vital role in several inflammatory diseases, however its role in the tumour microenvironment and the potential in cancer treatment is still unrevealed. Considering the potential of {beta}-lactams as HNE inhibitors, the present work describes the development of a synthetic strategy to obtain two different types (Type I and Type II) of quenched activity-based probes (qABPs), using a {beta}-lactam ring as a warhead and BODIPY-FL as a fluorophore. The two types differ in mechanism and relative position between the fluorophore and the quencher moiety. The qABPs synthesized presented IC50 values against HNE lower than 0.5 {micro}M, and high selectivity compared with homologous serine hydrolases. Type II qABPs showed a more efficient turn-on mechanism, and selectively targeted HNE in different cell lysates. The qABP 22 was internalized in U937 cells and in human neutrophils and successfully targeted HNE in both.

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