Box–Behnken Design Optimization of High-Pressure Processed Bitter Melon (Momordica charantia) Leaf Extract Enhancing Phytochemicals, Anticancer, and Anti-Inflammatory Activities
3
Issued Date
2026-06-01
Resource Type
ISSN
16616596
eISSN
14220067
Scopus ID
2-s2.0-105041571789
Journal Title
International Journal of Molecular Sciences
Volume
27
Issue
11
Rights Holder(s)
SCOPUS
Bibliographic Citation
International Journal of Molecular Sciences Vol.27 No.11 (2026)
Suggested Citation
Kulchim K., Braspaiboon S., Pitchakarn P., Imsumran A., Buacheen P., Laowanitwattana T., Temviriyanukul P., Chairatvit K., Wongnoppavich A. Box–Behnken Design Optimization of High-Pressure Processed Bitter Melon (Momordica charantia) Leaf Extract Enhancing Phytochemicals, Anticancer, and Anti-Inflammatory Activities. International Journal of Molecular Sciences Vol.27 No.11 (2026). doi:10.3390/ijms27114945 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/117402
Title
Box–Behnken Design Optimization of High-Pressure Processed Bitter Melon (Momordica charantia) Leaf Extract Enhancing Phytochemicals, Anticancer, and Anti-Inflammatory Activities
Corresponding Author(s)
Other Contributor(s)
Abstract
Bitter Melon Leaf Extract (BMLE) possesses potential anticancer and anti-inflammatory properties; however, conventional extraction methods restrict phytochemical yield and bioactivity. Here, we optimized extraction using High-Pressure Processing (HPP) with Box–Behnken Design (BBD) and Response Surface Methodology (RSM). The optimized extract (O-BMLE) demonstrated significantly higher total flavonoid content (27.7 vs. 8.7 mg RE/g) and FRAP antioxidant capacity (96.5 vs. 71.2 μmol TE/g) compared to conventional BMLE. Additionally, O-BMLE exhibited enhanced cytotoxicity (A549 IC<inf>50</inf>: 58.7 vs. 147 μg/mL) and selectivity (SI: 5.03 vs. 2.60) against A549, HepG2, and SKOV3 cancer cells while showing minimal effects on 3T3-L1 fibroblasts. In LPS-stimulated RAW264.7 macrophages, O-BMLE selectively inhibited JNK phosphorylation without affecting NF-κB phosphorylation, resulting in suppression of iNOS, COX-2, IL-1β, IL-6, and TNF-α expression as well as nitric oxide production. HPLC analysis revealed equivalent momordicine-I levels (~28 mg/g) between extracts. In contrast, HPLC-qTOF-MS profiling revealed that O-BMLE was enriched in stearidonic acid (66% increase in relative abundance), 4-hydroxybenzoic acid (19.5%), monolinolenin, 6-gingerol, and pedunculoside, which are compounds linked to JNK inhibition, antioxidant activity, and cytokine suppression. These results indicate that HPP-BBD/RSM optimization selectively modifies the bitter melon leaf metabolome, thereby enhancing anticancer and anti-inflammatory activities independently of momordicine-I content alone. O-BMLE may therefore serve as a promising candidate for the development of functional foods and nutraceuticals targeting inflammation-associated cancers.