Potential Anticancer Activity of the Furanocoumarin Derivative Xanthotoxin Isolated from Ammi majus L. Fruits: In Vitro and In Silico Studies

Abstract

Ammi majus L., an indigenous plant in Egypt, is widely used in traditional medicine due to its various pharmacological properties. We aimed to evaluate the anticancer properties of Ammi majus fruit methanol extract (AME) against liver cancer and to elucidate the active compound(s) and their mechanisms of action. Three fractions from AME (Hexane, CH2Cl2, and EtOAc) were tested for their anticancer activities against HepG2 cell line in vitro (cytotoxicity assay, cell cycle analysis, annexin V-FITC apoptosis assay, and autophagy efflux assay) and in silico (molecular docking). Among the AME fractions, CH2Cl2 fraction revealed the most potent cytotoxic activity. The structures of compounds isolated from the CH2Cl2 fraction were elucidated using 1H- and 13C-NMR and found that Compound 1 (xanthotoxin) has the strongest cytotoxic activity against HepG2 cells (IC50 6.9 ± 1.07 µg/mL). Treating HepG2 cells with 6.9 µg/mL of xanthotoxin induced significant changes in the DNA-cell cycle (increases in apoptotic pre-G1 and G2/M phases and a decrease in the S-phase). Xanthotoxin induced significant increase in Annexin-V-positive HepG2 cells both at the early and late stages of apoptosis, as well as a significant decrease in autophagic flux in cancer compared with control cells. In silico analysis of xanthotoxin against the DNA-relaxing enzyme topoisomease II (PDB code: 3QX3) revealed strong interaction with the key amino acid Asp479 in a similar fashion to that of the co-crystallized inhibitor (etoposide), implying that xanthotoxin has a potential of a broad-spectrum anticancer activity. Our results indicate that xanthotoxin exhibits anticancer effects with good biocompatibility toward normal human cells. Further studies are needed to optimize its antitumor efficacy, toxicity, solubility, and pharmacokinetics.

In Vivo Study the Cytogenetic Effect of Ammi majus Methanolic Extract on Mitotic Index, Micronucleus Formation and DNA Damage on Mitoxantrone Treated Albino Male Mice

“…The disparities between normal and cancer cells' reactions to xanthotoxin may be explained by one of the most distinguishing characteristics of cancer cells over those of normal cells 26 : increased expression of the relaxing enzyme topoisomerase II 27,28 . Finally, all these effects of plant are summarized as follow: ammi majus secondary metabolite possesses the capacity to stimulate the function of the immune system, inhibit the formation of DNA adduct with carcinogens, inhibit hormonal action and metabolic pathways associated with the development of cancer and enhance the phase I or II detoxification enzymes 29,30. Furthermore, investigations have shown that terpenoids may increase tumor latency and decrease tumor multiplicity 31 .…”

Xanthotoxin (8‐methoxypsoralen): A review of its chemistry, pharmacology, pharmacokinetics, and toxicity

“…8‐MOP can increase the content of phosphorylated Chk1 and decrease the proportion of phosphorylated cdc2 (Sumiyoshi, Sakanaka, Taniguchi, Baba, & Kimura, 2014). They assayed the cytotoxicity of XAT with HepG2 cells and showed that XAT had strong cytotoxic activity against HepG2 cells (IC 50 = 6.9 ± 1.07 μg/mL), then HepG2 cells were treated with 6.9 μg/mL of XAT and cell cycle was observed, which showed significant changes in cell cycle (increase in G1 and G2/M phases and decrease in s phase), XAT induced a significant increase in Annexin‐v‐positive HepG2 cells in the early and late stages of apoptosis compared to controls, along with a significant decrease in autophagy in cancer cells (Issa et al, 2022). Inflammatory signaling pathways are intimately related to cancer.…”

Two novel coumarin-based fluorescent probes for the detection of Cu2+ and biological applications

“…[22][23][24][25][26][27][28][29][30][31][32] Coumarin derivatives can also be used as fluorescence markers or imaging agents to identify tumor lesions and study the subcellular localization of drugs. [33][34][35][36][37][38][39][40][41][42][43][44] The development of a sensor with high selectivity and sensitivity to copper ions remains an important challenge. In this paper, two novel fluorescent probes were synthesized using coumarin with an aldehyde (CHO group) as the fluorophore.…”