Development, Characterisation, and Pharmacological Assessment of <i>Eclipta alba</i> Phytosomes as Potential Anti-cancer formulation

Background: Eclipta alba leaf extract has demonstrated promising anticancer potential; however, its clinical application is limited by poor bioavailability and rapid metabolism. Aim: This study aimed to enhance the therapeutic efficacy and bioavailability of Eclipta alba leaf extract by encapsulating it within phytosomes and evaluating its antiproliferative effects on 4T1 breast cancer cells, as well as its safety in vivo. Methods: Phytosomes were synthesised using the nanoprecipitation technique and characterised by dialysis membrane studies and Dynamic Light Scattering (DLS). Key parameters such as particle size, zeta potential, Polydispersity Index (PDI), encapsulation efficiency, and total phenolic content were evaluated. Drug release profiles were assessed, and in vitro cytotoxicity and antiproliferative activity were determined using the MTT assay. Acute toxicity was assessed in Swiss albino mice according to OECD guidelines. Results: The prepared phytosomes showed a particle size of 296 ± 0.29 nm, zeta potential of −40.1 ± 1.19 mV, and a PDI of 0.106 ± 0.002, indicating good stability and uniformity. The encapsulation efficiency was 90.32 ± 0.11%, and the total phenolic content was 50.81 ± 0.02 mg GAE/g. Drug release exhibited a biphasic pattern with sustained release following an initial burst. Phytosomes displayed higher antiproliferative activity (IC₅₀ = 7.73 ± 2.87 μg/mL, selectivity index > 3) against 4T1 cells compared to both the free extract (IC₅₀ = 212.9 ± 1.30 μg/mL) and doxorubicin (IC₅₀ = 68.35 ± 3.508 μg/mL), with statistically significant results (p<0.001). Acute toxicity studies confirmed safety up to 2000 mg/kg. Conclusion: Phytosomes loaded with Eclipta alba leaf extract significantly enhance therapeutic efficacy and show a favourable safety profile, suggesting their strong potential as a novel drug delivery system for breast cancer treatment. Major Findings: Phytosomes loaded with Eclipta alba leaf extract demonstrated enhanced antiproliferative activity and reduced cytotoxicity against 4T1 breast cancer cells compared to the free extract and doxorubicin. They were well tolerated in vivo at doses below 2000 mg/kg, indicating their potential as a safe and effective drug delivery system for breast cancer therapy.