Anti‐Inflammatory and Cytotoxic Assessment of Alkaloid Rich Fraction From Sarcostigma kleinii Wight & Arn. With Pharmaco‐Informatic Studies

ABSTRACT Inflammation is an intricate physiological process that causes many illnesses. S. kleinii has been traditionally used as food and medicine, especially for rheumatic diseases associated with inflammation. However, its anti‐inflammatory properties remain scientifically unverified. A systematic protocol for the extraction and characterization of alkaloid‐rich fraction from S. kleinii fruits (SkAF) was developed. In vitro enzyme inhibitory and MTT assays are used to evaluate their anti‐inflammatory and cytotoxicity potential. The UV‐visible and FTIR methods characterized the SkAF, while marker‐based LC‐MS/MS analysis identified CPT (m/z 349) as an active alkaloid present in the fraction. The SkAF did not adversely affect RAW 264.7 and L929 cell lines. At the highest dose (100 µg/mL), cell viability remained high at 92.48% and 94.47%. In addition, HPTLC fingerprinting estimated 45.32 ng/mL of CPT in the SkAF. Molecular docking was performed between CPT and different sets of LOX‐1, COX‐2, 5‐LOX, and NF‐Kappa B proteins, and CPT was found to be more effective against 5‐LOX (‐9.7 kcal/mol) and COX‐2 (‐9.5 kcal/mol) proteins. SkAF possessed significant anti‐inflammatory activity by effectively inhibiting COX‐2 and 5‐LOX without inducing any cytotoxic effects on L929 mouse fibroblast cells. This study provides scientific validation for the traditional application of this fruit in treating inflammation.