Pyran-annulated derivatives synthesized via multi-component reactions as multi-target agents with anti-inflammatory, anti-melanogenesis, and anti-cancer activities

Pyran-annulated derivatives are multifunctional scaffolds that exhibit diverse biological activities, including anti-inflammatory, anti-melanogenesis, and anti-cancer effects. This study aimed to synthesize a series of pyran-annulated derivatives and evaluate their biological properties in vitro, to identify potential multi-target therapeutic candidates. The synthesized compounds were screened for anti-inflammatory activity by measuring nitric oxide (NO) inhibition in lipopolysaccharide (LPS)-stimulated J774A.1 macrophages. Anti-melanogenesis activity was evaluated in B16F10 melanoma cells, while cytotoxicity was assessed against MDA-MB-231 triple-negative breast cancer cells. Western blot assays were conducted to examine the expression of inflammation and melanogenesis-related proteins, while molecular docking was performed to investigate binding affinity to the cyclooxygenase-2 (COX-2) active site. In addition, wound-healing assays were carried out to assess the impact of cancer cell migration. Among the tested compounds, A05 emerged as the most active compound, displaying potent multi-target effects. It showed an IC₅₀ value of 10.2 µM for NO inhibition, surpassing indomethacin (IC₅₀ = 44.5 µM), 13.3 µM for melanin inhibition, and 24.3 µM for cytotoxicity against MDA-MB-231 cells. Western blot analysis confirmed that A05 significantly downregulated COX-2, iNOS, and TNF-α in J774A.1 macrophages, while suppressing MITF and TRP-1 expression in B16F10 cells. Molecular docking results revealed strong binding of A05 to the COX-2 active site, supporting its anti-inflammatory action. Furthermore, wound-healing assays demonstrated that A05 effectively inhibited cancer cell migration. These findings highlight A05 as a promising lead compound with combined anti-inflammatory, anti-melanogenesis, and anti-cancer properties. Its ability to simultaneously modulate multiple biological pathways underscores its potential as a candidate for the development of multi-target therapeutic agents.