Integrated in silico and in vitro evaluation of cocoa pod husk as potential anticancer agents against hepatocellular carcinoma.

BACKGROUND

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide, with limited efficacy of current therapies and frequent drug resistance. Cocoa pod husk (CPH), an abundant agro-industrial by-product, is rich in bioactive compounds but remains underutilized. This study investigated the anti-HCC potential of CPH using integrated green extraction, in silico prediction, and in vitro validation.

METHODS

CPH extracts were prepared via ultrasound-assisted extraction (CPH-UE) and enzymatic extraction (CPH-EE). Phytochemical profiling was conducted using UHPLC-ESI-HRMS/MS. In silico analyses included structure-activity relationship prediction, ADMET profiling, network pharmacology, and molecular docking against key HCC targets (EGFR, PIK3CA, and SRC). Anticancer activity was evaluated in HepG2 and Huh7 cell lines using MTT assays, with THLE-2 normal liver cells as control.

RESULTS

CPH-UE was enriched in polyphenols (catechin, epicatechin, quercetin), whereas CPH-EE contained bioactive peptides and procyanidin B2. In silico analyses highlighted EGFR, PIK3CA, and SRC as central targets, with favorable binding affinities for major polyphenols. In vitro assays demonstrated dose-dependent antiproliferative effects, with CPH-UE showing lower IC₅₀ values and better selectivity toward HCC cells than CPH-EE.

CONCLUSION

Cocoa pod husk exhibits promising anti-HCC activity through multi-component, multi-target mechanisms. Ultrasound-assisted extraction yields polyphenol-rich extracts with superior anticancer potency, supporting CPH valorization as a sustainable source of functional anticancer agents.