Exploring Anticancer Potential of Aqueous, Hydroethanolic and Methanolic Extract of an Age-old <i>Unani</i> Formulation

Background: Cancer poses a foremost public health challenge worldwide. Several cancer types are preventable and treatable, provided that cancer is diagnosed at an early stage. The derivatives of plant metabolites such as camptothecin, podophyllotoxin, vinca, and others have been a crucial source of numerous clinically effective anti-cancer agents. Aim: The present study was conducted to evaluate the cytotoxic potential of a traditional Unani formulation against cancer cell lines. Methods: A compound Polyherbal Unani Formulation (PUF) mentioned in the Unani text Kamil-Us-Sana to evacuate Khilt-e-Sawdā’ in the chapter of Saraṭān (Cancer) was selected to screen its anti-cancer potential against A-549, HeLa, MCF-7 and B16F10 cells. Aqueous, hydroethanolic (1:1), and methanolic extract of PUF was prepared and tested. Results: Total phenolics and flavonoids were detected in significant quantities in PUF. PUF and its ingredients exhibited powerful anti-oxidant potential in DPPH, ABTS, and FRAP assay. Methanolic extract of PUF was found most potent cytotoxic (IC50 value of 28.50±1.53 μg/mL) in B16F10 cells, almost equal to positive control doxorubicin (IC50 24.62±0.19 μg/mL). PUF also increased the caspase-3 activity in HeLa and A-549 cells, suggesting apoptosis induction. The cytotoxic activity of PUF may be attributed to the effect of its ingredients in MCF-7 cells and other cancer cell lines. Conclusion: Present findings validate the traditional claim mentioned in the Unani text and open up a new avenue for further research to explore molecular mechanisms and development of this Unani formulation for effective cancer management. Major Findings: Present study demonstrated the cytotoxic potential of a traditional polyherbal Unani formulation with IC50 values ranging from 28.50±1.53 μg/mL to 207.86±1.24 μg/mL in HeLa, A-549, MCF-7, and B16F10 cancer cells. The methanolic extract of PUF exhibited greater cytotoxic potential compared to hydro-ethanolic and aqueous extract. The cytotoxic effects of PUF are likely due to its ingredients, which have been reported to have cytotoxic activity in other cancer cell lines.