# Integrative analysis of Trichosanthes kirilowii maxim formula granules’ anti-triple-negative breast cancer mechanism via network pharmacology, metabolomics, and molecular pharmacology

## TL;DR

This study explores how Trichosanthes kirilowii granules may treat triple-negative breast cancer by analyzing their effects on cell cycles, apoptosis, and key signaling pathways.

## Contribution

The study integrates network pharmacology, metabolomics, and molecular experiments to reveal multi-target mechanisms of a herbal formula against TNBC.

## Key findings

- TKM promotes apoptosis and induces G2/M-phase cell-cycle arrest in TNBC cells.
- Schisandrin binds to key targets like MYC and AKT1 with strong docking performance.
- TKM regulates amino acid metabolism and signaling pathways like PI3K/AKT and Wnt/β-catenin.

## Abstract

Triple-negative breast cancer (TNBC) urgently needs effective therapies due to limited targeted options and unfavorable outcomes. We investigated Trichosanthes kirilowii Maxim formula granules (TKM) using integrated network pharmacology, metabolomics, and molecular pharmacology to clarify potential multi-target mechanisms relevant to TNBC.

Liquid chromatography coupled with mass spectrometry was employed to identify the components of TKM formula granules. Network pharmacology-based prediction was used to uncover potential mechanisms by which TKM counteracts TNBC. Potential targets were identified, and pathway enrichment analysis was performed. Subsequently, TNBC cells and 4T1 tumor-bearing mice were used to verify the molecular mechanisms of TKM.

We identified 151 active compounds in TKM. Through network pharmacology analysis, 214 TNBC-related targets were found, with 28 core targets, including cell cycle and apoptosis regulators MYC, TP53, AKT1, CCND1, CASP3, PIK3CA, BCL2L1, and CDC42. The compound-target-pathway-disease network showed that schisandrin binds to many treatment targets with satisfactory docking performance, especially for MYC and AKT1. Experimentally, TKM was found to significantly promote apoptosis and induce G2/M-phase cell-cycle arrest in MDA-MB-231 cells. Western blot analysis showed that TKM suppressed PI3K/AKT and Wnt/β-catenin signaling pathways. Surface plasmon resonance experiment revealed that schisandrin binds to recombinant AKT1 with an equilibrium dissociation constant (K_D) of 1.525 × 10−4 M. According to untargeted metabolomics results, TKM can regulate amino acid, glycine, serine, and threonine metabolism, mineral absorption, protein digestion and absorption, central carbon metabolism, and arginine biosynthesis to exert therapeutic effects on TNBC. All findings were consistent with predicted targets and pathways.

This study comprehensively explores the multi-target mechanisms of TKM against TNBC using network pharmacology, molecular pharmacology, and metabolomics approaches. These findings provide a foundation for future mechanistic investigations and may support the further preclinical development of TKM-based strategies for TNBC.