In Vitro and In Silico Evaluation of Cladophora sp. as an Anti-Inflammatory Agent via COX-2 (Cyclooxygenase-2) Inhibition

Department of Medicine, Universitas Islam Malang, Malang 65144, Indonesia

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Abstract

Cladophora sp. is known to contain chemical constituents with pharmacological properties. The secondary metabolites present in Cladophora sp. include alkaloids, phenolic compounds, saponins, and terpenoids, and it has been reported to exhibit anti-inflammatory activity. To evaluate its potential as an anti-inflammatory agent, a study was conducted using in silico and in vitro approaches. The in silico analysis involved screening the physicochemical characteristics and pharmacokinetic profiles of the compounds. Meanwhile, the in vitro analysis was performed using a bovine serum albumin (BSA) protein denaturation assay. Docking studies with the receptor protein showed that compounds from the ethanol extract of Cladophora with lower binding affinity to COX-II (PDB ID: 5kir) compared to the control drug rofecoxib were (2R)-5-hydroxy-7-methoxy-2-phenyl-3,4-dihydro-2H-1-benzopyran-4-one (-9 kcal/mol) and pinocembrin (-8.9 kcal/mol). The compound (2R)-5-hydroxy-7-methoxy-2-phenyl-3,4-dihydro-2H-1-benzopyran-4-one showed 100% similarity in amino acid residues with the control, forming hydrogen bonds at His90 and Arg513. The in vitro anti-inflammatory assay produced a linear regression equation of y = 352.52x - 1506.3 with an r² value of 0.9179, and an IC₅₀ value of 82.664 ppm, indicating strong anti-inflammatory activity. Further studies are recommended to isolate (2R)-5-hydroxy-7-methoxy-2-phenyl-3,4-dihydro-2H-1-benzopyran-4-one for subsequent in vitro and in vivo antiinflammatory evaluations.