A New Method for Purifying Brazilin from Lignum Sappan – Cytotoxic and Anti‑Proliferative Effects in Cell Lines and Improved Survival in Mice Bearing Urinary Bladder Carcinoma

Objectives: The present study describes a methodology for extracting and obtaining high purity brazilin from Lignum sappan and evaluates its effects on urinary bladder carcinoma, in vitro and in vivo. Materials and Methods: The brazilin was extracted from Lignum sappan, purified and identified. The purified brazilin was assessed in vitro for its effects on tumor cell viability and growth using human (T24) and mouse urinary bladder (BTT) cancer cell lines. The role of brazilin in the inhibition of the cell cycle was determined by flow cytometry. The in vivo activity of brazilin was assessed based on the survival of tumor‑bearing mice. Results: Brazilin was purified of 99.10% purity and was bioactive as it significantly (P < 0.05) inhibited the growth of T24 and BTT cells. The IC50 was 10.08 µg/ml for T24 cells and 8.76 µg/ml for BTT cells after 48 h of treatment. Brazilin blocked the cell cycle progression in the G2 phase. Brazilin (100, 200 and 300 mg/kg) significantly (P < 0.05) increased the survival of BTT tumor‑bearing T739 mice and T24 tumor‑bearing BALB/c nude mice. Conclusion: The methodology described for the extraction of brazilin from Lignum sappan yields high purity and bioactive brazilin. The extracted brazilin exerts marked inhibitory effects on urinary bladder cancer cells in vitro and in vivo.