Integrated Network Pharmacology, Molecular Modeling, LC-MS Profiling, and Semisynthetic Approach for the Roots of Rubia tinctorum L. Metabolites in Cancer Treatment

Sep 23, 2026

Rubia tinctorum L. metabolites are investigated for anticancer efficacy using a multilevel strategy combining network pharmacology, molecular docking, and in vitro cytotoxicity. Network analysis highlights acacetin, alizarin, anthragallol, 2-hydroxyanthraquinone, and xanthopurpurin as key constituents, with PLCG1, BCL2, CYP1B1, NSD2, and ESR2 as enriched targets. Docking links acacetin and alizarin to lowest binding energies, and anthragallol to stabilized interactions, followed by LC/MS quantification and a semisynthetic alizarin route yielding 2-methyl alizarin with the strongest HepG2 activity.

LanguageEnglish

CategoryHealthcare

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Pages19

Reading Time48 min

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Abstract

Introduction

This article is licensed under CC-BY-NC-ND 4.0 [http://pubs.acs.org/journal/acsodf](http://pubs.acs.org/journal/acsodf) Article Integrated Network Pharmacology, Molecular Modeling, LC−MS Profiling, and Semisynthetic Approach for the Roots of Rubia tinctorum L. Metabolites in Cancer Treatment Alaa A. El-Banna,‡‡ Enas E. Eltamany,‡‡ Asmaa S. A. Yassen, Ahmed Lotfy, Aya H. H. El-Tanahy, Jihan M. Badr, * Mardi M. Algandaby, Samar S. Murshid, Sameh S. Elhady, and Reda F. A. Abdelhameed * Cite This: ACS Omega 2025, 10, 13027−13045 Read Online | | | | | | | | --- | --- | --- | --- | --- | --- | | ACCESS | Metrics & More | | Article Recommendations | | *sı Supporting Information | ABSTRACT: Rubia [tinctorum](tinctorum L. is)[ L. is](tinctorum L. is) one of the most widely used plants in folk medicine, with many reported pharmacological activities. One of these valuable activities is its anticancer efficacy. The aim of this study is to explore the multilevel mechanisms of R. tinctorum metabolites in cancer treatment using network pharmacology, together with molecular docking and in vitro studies. The network pharmacology analysis enabled us to reveal the hit anticancer R. tinctorum constituents, which were found to beacacetin, alizarin, anthragallol, 2-hydroxyanthraquinone, and xanthopurpurin. The most enriched cancer-linked target genes were PLCG1, BCL2, CYP1B1, NSD2, and ESR2. The pathways that were mostly involved in the anticancer mechanism of R. tinctorum metabolites were found to be metabolic pathways as well as pathways in cancer and apoptosis. Molecular docking of the identified hit anticancer constituents on the active sites of the most enriched genes unveiled that acacetin and alizarin possessed the lowest binding energies on the active sites of NSD2 and BCL2, respectively. While anthragallol showed the most stabilized interaction on the active sites of PLCG1, CYP1B1, and ESR2. Consequently, R. tinctorum extracts were evaluated for their in vitro cytotoxicity on a panel of cancerous cells. Among the tested R. tinctorum extracts, the chloroform extract was the strongest one with an IC50 = 3.987 μg/mL on the MCF-7 breast cancer cell line. Consequently, it was subjected to chromatographic separation and purification to isolate its major components with reported anticancer activity (scopoletin, rubiadin, chrysophanic acid, alizarin, purpurin, nor-damnacanthal, emodin, and rutin). Alizarin and purpurin constituted the main anthraquinones in R. tinctorum. Thus, they were quantified using LC/MS analysis. Moreover, asemisynthetic approach of alizarin toward the enhancement of its anticancer effect on the tested cancer cells was attained. Among the synthesized compounds, 2-methyl alizarin was the most active one with an IC50 = 8.878 μg/mL against the HepG2 cell line. This study provides deep insights into the anticancer mechanisms of R. tinctorum metabolites for the first time using network pharmacology and valorizes their significance as valuable anticancer agents. 1. INTRODUCTION Cancer is a serious health problem that is characterized by uncontrollable body cell growth that can pervade throughout the body.1 It is the second main cause of death globally, behind cardiovascular ailments.2 In the past decade, there have been 10 million cancer-related deaths and 20 million new instances of cancer worldwide. By 2040, it is anticipated that there will be roughly 30 million additional instances of cancer worldwide, with the largest increases occurring in middle-and low-income nations, if no new steps are taken to control and prevent cancer.3 For many years, patients with cancer had only a limited number of treatment options, which included surgery, radiation therapy, and chemotherapy, either individually or in combination.4 Although these treatment methods were somewhat effective, they exhibited many adverse side effects and problems such as the drug nonselectivity and resistance, in addition to fatigue

FAQs

What methods were used to study Rubia tinctorum L. metabolites for cancer treatment?

The study combines network pharmacology, molecular docking, and in vitro cytotoxicity assays, followed by chromatographic separation, LC/MS quantification, and a semisynthetic approach using alizarin.

Which anticancer-related constituents and targets were identified by network pharmacology?

Key hit constituents include acacetin, alizarin, anthragallol, 2-hydroxyanthraquinone, and xanthopurpurin. Enriched cancer-linked target genes reported are PLCG1, BCL2, CYP1B1, NSD2, and ESR2.

What were the strongest in vitro cytotoxicity results and the key semisynthetic outcome?

The chloroform extract showed the strongest activity with IC50 = 3.987 μg/mL against MCF-7. The semisynthetic 2-methyl alizarin exhibited the strongest activity among synthesized compounds with IC50 = 8.878 μg/mL against HepG2.

Sep 23, 2026

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