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<p>Breast cancer remains a major global health challenge, necessitating the discovery of new therapeutic agents with enhanced efficacy and minimal side effects. This study explores the potential of <i>Timonius flavescens</i> (Jacq) Baker leaf extract, rich in polyphenols, as an anti-breast cancer agent. A green solvent system using Natural Deep Eutectic Solvent (NADES), composed of choline chloride and glucose, was optimized via Central Composite Design (CCD) to maximize the extraction of phenolic and flavonoid compounds. At an MoR of 2.084 and water content of 29.581%, the optimized extract yielded a superior total phenolic content (TPC) of 66.54 ± 0.586&#xa0;mg GAE/g and total flavonoid content (TFC) of 28.89 ± 0.26&#xa0;mg QE/g, compared to conventional ethanol extraction. In addition, the optimized extract revealed higher antioxidant activity based on DPPH assays. In vitro cytotoxicity assessments against MCF-7, T47D, and 4T1 breast cancer cell lines revealed a dose-dependent reduction in cell viability, with IC<sub>50</sub> values of 116.20&#xa0;µg/mL, 258.64&#xa0;µg/mL, and 235.03&#xa0;µg/mL, respectively. The extract induced G0-G1 cell cycle arrest and apoptosis via p53 upregulation and cyclin D1 downregulation, as confirmed by flow cytometry and immunocytochemistry. Additionally, an acute toxicity evaluation in mice indicated the extract’s safety, showing no significant adverse effects on hematological and biochemical parameters or organ histology. These findings highlight the therapeutic potential of <i>T. flavescens</i> leaf extract as a natural anti-breast cancer agent, supporting its further investigation in preclinical and clinical studies. Additionally, this study underscores the efficacy of NADES-based extraction in enhancing bioactive compound recovery, promoting green chemistry in pharmaceutical research.</p> Graphical abstract <p></p>

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The potential of natural deep eutectic solvent in polyphenol extraction of Timonius flavescens (Jacq) Baker as an anti-breast cancer agent and its toxicity evaluation

  • 期刊论文

  • 发表日期：

    2025年5月27日

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  • Muhammad Fauzan Lubis,
  • Retno Murwanti,
  • Poppy Anjelisa Zaitun Hasibuan,
  • Sumaiyah Sumaiyah,
  • Nur Aira Juwita,
  • Ana Yulyana,
  • Ririn Astyka,
  • Sri Yuliasmi,
  • Lokot Donna Lubis,
  • Wardiyah Daulay,
  • Khairani Fitri

摘要

Breast cancer remains a major global health challenge, necessitating the discovery of new therapeutic agents with enhanced efficacy and minimal side effects. This study explores the potential of Timonius flavescens (Jacq) Baker leaf extract, rich in polyphenols, as an anti-breast cancer agent. A green solvent system using Natural Deep Eutectic Solvent (NADES), composed of choline chloride and glucose, was optimized via Central Composite Design (CCD) to maximize the extraction of phenolic and flavonoid compounds. At an MoR of 2.084 and water content of 29.581%, the optimized extract yielded a superior total phenolic content (TPC) of 66.54 ± 0.586 mg GAE/g and total flavonoid content (TFC) of 28.89 ± 0.26 mg QE/g, compared to conventional ethanol extraction. In addition, the optimized extract revealed higher antioxidant activity based on DPPH assays. In vitro cytotoxicity assessments against MCF-7, T47D, and 4T1 breast cancer cell lines revealed a dose-dependent reduction in cell viability, with IC50 values of 116.20 µg/mL, 258.64 µg/mL, and 235.03 µg/mL, respectively. The extract induced G0-G1 cell cycle arrest and apoptosis via p53 upregulation and cyclin D1 downregulation, as confirmed by flow cytometry and immunocytochemistry. Additionally, an acute toxicity evaluation in mice indicated the extract’s safety, showing no significant adverse effects on hematological and biochemical parameters or organ histology. These findings highlight the therapeutic potential of T. flavescens leaf extract as a natural anti-breast cancer agent, supporting its further investigation in preclinical and clinical studies. Additionally, this study underscores the efficacy of NADES-based extraction in enhancing bioactive compound recovery, promoting green chemistry in pharmaceutical research.

Graphical abstract