Development And In Vitro Analysis Of Effervescent Tablet Using Swertia Chirata, Beetroot And Triphala As Chemopreventive Agents

Abstract

Background: Natural products are promising chemopreventive agents against cancer showing multi-target activity with less toxicity compared to conventional chemotherapy. Swertia chirata, beetroot, and triphala have well-known anticancer as well as antioxidant properties; however, formulation of these into patient-compliant dosage forms is quite unexplored.

Objective: To develop and evaluate the effervescent tablets of Swertia chirata, beetroot, and triphala extracts, as well as to systematically assess their cytotoxic activity, cell cycle modulatory properties and free radicals scavenging ability against A549 non-small cell lung cancer (NSCLC) cells.

Materials and methods: A standardized base consisting of citric acid, sodium bicarbonate, mannitol, and pharmaceutical excipients was used to prepare seven effervescent tablet formulations (F1-F7). Ethanol (70%) extracts were prepared for each individual botanical and their combination. Based on sensory evaluation, the optimized formulation (F6) was selected for biological assessment. In- vitro cytotoxicity was assessed using MTT assay (0–1000 µg/mL), cell cycle distribution was analyzed via flow cytometry, and antioxidant potential by DPPH radical scavenging assay. Paclitaxel was used as a positive control in an A549 lung adenocarcinoma cell line-based model.

Results: All extracts showed dose-dependent cytotoxicity against A549 cells. Cell cycle profile demonstrated that extracts S1 (Swertia chirata), S2 (beetroot) and S3 (triphala) resulted in significant S-phase arrest, with extract S3 showing the strongest effect at 25.89 µg/mL IC₅₀, resulting in approximately 49% accumulation of cells within this phase. In contrast, the equal-ratio combination (S4) and effervescent tablet formulation (F6) were significantly less effective as evaluated by cell cycle arrest activity, with cells continuing through phases similar to untreated controls. DPPH assay showed IC₅₀ values of 6.197 µg/mL for S3 (triphala), 16.53 µg/mL for S4 (combination), 151.3 µg/mL for F6 (tablet), 381.1 µg/mL for S1 (Swertia chirata), and >500 µg/mL for aptowin5490 (beetroot) as compared to 4.521µ g /mL ascorbic acid standard. Microscopic analysis (100×) at 100 µg/mL and 1000 µg/mL of concentrations demonstrated characteristic cytotoxic morphological changes such as cell shrinkage, blebbing, and reduced cell density.

Conclusion: Triphala showed more cytotoxic and cell cycle arrest activities compared to other botanical extracts against the A549 cells. Yet, the prepared effervescent tablet (F6) showed significantly reduced biological activity compared to crude extracts, indicating possible antagonistic interactions or decreased bioavailability during pharmaceutical processing. These results emphasise the necessity for formulation development to maintain the stability and bioavailability of bioactive compounds. Protective formulation approaches, alternative delivery systems, and pharmacokinetics optimization studies can be needed in the future to assure that these natural agents would still retain their chemopreventive potential.