Exploring the anti-asthmatic effects of HMM: OVA-induced murine model and multi-target in silico analysis

Asthma is a widespread chronic inflammatory condition of the airways, often triggered by allergens such as ovalbumin. This study aimed to evaluate the potential anti-asthmatic effects of a synthetic isoxazolone derivative, (Z)-4-(4-hydroxy-3-methoxybenzylidene)-3-methylisoxazole-5(4H)-one (HMM), in a mouse model of allergic asthma. Molecular docking was carried out to explore the HMM interaction with asthma-associated proteins, providing insights into its potential therapeutic targets. Thirty-six albino mice were randomly divided into six groups: negative control, positive control, standard drug group (methylprednisolone, 15 mg/kg), and three groups receiving HMM at doses of 3.2, 6.3, and 12.6 mg/kg. Mice were sensitized with ovalbumin on days 0 and 14, followed by intranasal exposure from days 15 to 21 to induce allergic airway inflammation. Blood and bronchoalveolar lavage fluid (BALF) were collected to measure total and differential leukocyte counts. Cytokine expression (IL-4, IL-5) and aquaporins (AQP-1, AQP-5) were quantified via real-time PCR. HMM-treated groups showed reduced inflammation, lowered Th2 cytokine levels, and improved lung water transport markers. Computational studies supported these findings, revealing strong binding affinities between HMM and TNF-α (- 6.07 kcal/mol), AKT1 (- 6.32 kcal/mol), and COX-2 (- 5.87 kcal/mol), forming stable hydrogen-bond interactions with key active-site residues. These results suggest HMM holds promise as a potential anti-asthmatic agent.