Thanks to a multidisciplinary approach that included the participation of experts in computational biology, in the study of proteins and lipids and in the use of experimental models of cystic fibrosis (CF), the researchers have identified new inhibitors of both human and fungal sphingosine-1-phosphate lyase (SPL), potentially able of performing the dual anti-inflammatory and antimicrobial activity in CF. During the FFC#16/2020 pilot project, the researchers have developed the methodology necessary to achieve the main objective, which was then applied in the FFC#19/2021 extension. In particular, 50 potential inhibitors were identified by computational analyses starting from the screening of more than 200,000 molecules. The researchers have then obtained the purified forms of human and fungal SPL with a dual purpose: i) obtain the three-dimensional structure of the two proteins to verify and optimize the computational analysis; ii) test the inhibitory activity of the compounds identified by computational analysis in order to select the most promising compounds. As a result, 4 compounds were selected. Due to their poor solubility, which prevented their analysis in in vitro models, the researchers are currently developing a powder formulation to be administered directly to the CF mouse model. In parallel, the researchers have performed an analysis of the lipid composition of the lung of CF mice, as a prerequisite for evaluating any “off target” effects of the compounds that could impact their safety. In conclusion, the experiments allowed not only to confirm the potential efficacy of human and fungal SPL inhibitors in CF, but also to identify candidates for subsequent optimization for drug development. Further safety and efficacy studies, with the models already developed, will be necessary in this bench-to-bedside translational effort.