Bronchorelaxant and anti-inflammatory profile of ensifentrine alone and combined with inhaled medications in lipopolysaccharide-challenged human airways from chronic obstructive pulmonary disease donors

The ENHANCE trials demonstrated that ensifentrine, an inhaled dual phosphodiesterase 3/4 inhibitor, offers additional benefits in chronic obstructive pulmonary disease (COPD). Its bifunctional activity warrants pharmacological investigation to elucidate mechanisms behind lung function improvements and exacerbation reduction. Human bronchial tissues from COPD donors were examined after lipopolysaccharide challenge. The effects of ensifentrine on bronchial relaxation were evaluated in response to carbachol and electrical field stimulation (EFS), also in combination with long-acting muscarinic antagonist (LAMA), long-acting β 2 -adredoreceptor agonists (LABA), and inhaled corticosteroids (ICS). Anti-inflammatory effects were assessed by profiling cytokines, alarmins, and growth factors. Drug interactions were analyzed using the Bliss-Loewe Uncertainty Range (BLUR) model. Ensifentrine induced concentration-dependent relaxation to carbachol in both epithelium-intact and epithelium-denuded tissues (E max 81.97±6.17%, pEC 50 4.86±0.17) and inhibited EFS-induced contractility from 0.1 μM, with maximum reduction of 29.01±2.50%. When combined with a LAMA, dual LABA/LAMA, or triple ICS/LABA/LAMA combinations, ensifentrine showed synergistic bronchorelaxant effect (overall increase in relaxation 20.47±0.39%), leading to near-complete relaxation (>90%) at higher concentrations. At lower concentrations, ensifentrine reduced inflammation by 84.34±8.69% in medium bronchi and 64.75±8.96% in small airways; at the higher concentrations, inflammation was almost abolished (>90%), comparable to roflumilast. Drug combinations showed synergistic anti-inflammatory interactions in 31.43% of cases, mainly affecting interleukin (IL)-10, IL-25, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor‐α. This study supports ensifentrine as an adjunct in COPD therapy, due to its dual bronchodilator and anti-inflammatory actions. Further research is warranted to optimize combination strategies to maximize clinical benefits and to identify additional activities in the target tissue.