# Metabolomics-guided identification of bioactive phytometabolites from South African plants targeting neuroblastoma

## TL;DR

This study identifies bioactive compounds from two South African plants that show potential in fighting neuroblastoma, a deadly childhood cancer.

## Contribution

The study introduces an integrated metabolomics and mitochondrial assay approach to discover phytometabolites with neuroblastoma-targeting potential.

## Key findings

- Acorus calamus and Lippia javanica extracts inhibited SH-SY5Y cell viability with IC50 values of 0.2886 and 0.3066 μg/μL, respectively.
- Metabolomic profiling revealed distinct phytochemical signatures, including flavonoids and alkaloids, with significant species-level differentiation.
- Key metabolites showed favorable ADME properties and blood-brain barrier permeability, suggesting therapeutic potential.

## Abstract

Neuroblastoma constitutes a solid tumor in pediatric populations, characterized by a dismal prognosis and a scarcity of effective therapeutic interventions. Medicinal flora from South Africa represents valuable sources of bioactive phytometabolites with potential relevance to neuroblastoma. This study employed an integrated workflow merging untargeted UPLC-MS/MS metabolomics, mitochondrial functional assays, and in silico absorption, distribution, metabolism, and excretion (ADME) prediction to systematically identify bioactive metabolites from Acorus calamus and Lippia javanica with activity against SH-SY5Y neuroblastoma cells. Cytotoxic effects were quantified utilizing the CCK-8 assay, while mitochondrial membrane potential (ΔΨm) was conducted through JC-1 flow cytometry. Untargeted UPLC-MS/MS profiling yielded metabolomic fingerprints, through PCA, PLS-DA, and OPLS-DA. ADME and drug-likeness were predicted using SWISSADME. Both plant extracts exhibited dose-dependent inhibition of SH-SY5Y cell viability, with IC50 values determined at 0.2886 μg/μL for A. calamus and 0.3066 μg/μL for L. javanica. The ΔΨm assessment indicated enhanced mitochondrial polarization (68.2% and 65.4% compared to 58.8% in untreated controls), implying modulation of mitochondrial functional status. Metabolomic profiling unveiled distinct phytochemical signatures, including flavonoids, phenolics, jasmonates, and alkaloids, exhibiting significant species-level differentiation (F = 936.71, R

2
= 0.989, p = 0.005). Notable metabolites such as isopropyl β-glucoside, 6β-hydroxymethandienone, and 7-epi-12-hydroxyjasmonic acid demonstrated favorable ADME characteristics and permeability across the blood-brain barrier. This investigation elucidates that A. calamus and L. javanica possess potential efficacy against neuroblastoma, underscoring the translational potential of African medicinal flora in pediatric oncology and necessitating further preclinical exploration.