Identification of HMOX-1-Targeting Natural Compounds in Camellia nitidissima Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches

Lingqiu Zhang; Fan Zhang; Haimei Liang; Xiangling Qin; Chunmei Liang; Manlu Zhong; Yuemi Mo; Jinling Xie; Xiaotao Hou; Jiagang Deng; Erwei Hao; Zhengcai Du

doi:10.3390/ph18060824

Identification of HMOX-1-Targeting Natural Compounds in Camellia nitidissima Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches

Pharmaceuticals (Basel). 2025 May 30;18(6):824. doi: 10.3390/ph18060824.

Authors

Lingqiu Zhang^{1

2}, Fan Zhang^{1

2

3}, Haimei Liang^{1

2}, Xiangling Qin^{1

2}, Chunmei Liang^{1

2}, Manlu Zhong^{1

2}, Yuemi Mo^{1

2}, Jinling Xie^{1

2}, Xiaotao Hou^{1

2}, Jiagang Deng^{1

2

3}, Erwei Hao^{1

2}, Zhengcai Du^{1

2}

Affiliations

¹ Guangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, China.
² Guangxi Key Laboratory of TCM Formulas Theory and Transformation for Damp Diseases, Institute of Traditional Chinese and Zhuang-Yao Ethnic Medicine, Guangxi University of Chinese Medicine, Nanning 530200, China.
³ International Zhuang Medical Hospital, Guangxi University of Chinese Medicine, Nanning 530000, China.

Abstract

Background/Objectives:Camellia nitidissima Chi (C. nitidissima), a traditional Chinese "food and medicine homology" plant, has demonstrated potential anti-tumor properties. However, its mechanisms of anti-lung cancer activity via ferroptosis remain unclear. This study aimed to construct an integrated research system of "natural product extraction-purification, bioactivity evaluation, and computational drug screening" to explore the bioactive compounds in C. nitidissima leaves targeting HMOX-1-mediated ferroptosis and their anti-lung cancer mechanisms. Methods: Active fractions were prepared using ethanol extraction combined with polyamide column chromatography. The anti-lung cancer activity was evaluated using the NCI-H1975 cell model. Ferroptosis was verified via transmission electron microscopy (TEM), biochemical indicators, a PCR Array, and immunofluorescence. The bioactive compounds were identified using UPLC-Q Exactive MS, and their binding affinity to HMOX-1 was evaluated via molecular docking and dynamics simulations, followed by cellular validation. Results: The 95% F1 fraction from the extracts of C. nitidissima leaves exhibited the strongest anti-lung cancer activity, which could be significantly reversed by Ferrostatin-1. Furthermore, it induced typical ferroptosis-related structural damage in mitochondria, including shrinkage and a reduction in size, increased membrane density, and a reduction or even the disappearance of cristae structures. At the molecular level, this fraction significantly increased the levels of oxidative stress markers (ROS↑, MDA↑, Fe²⁺↑, and GSH↓) and upregulated the expression of key ferroptosis-related genes, including HMOX-1, CHAC1, and NOX1. Using UPLC-Q Exactive MS combined with computational simulation methods, four bioactive compounds with high affinity for HMOX1 were successfully identified, including isochlorogenic acid A (-8.4 kcal/mol), isochlorogenic acid C (-8.4 kcal/mol), apigenin (-7.8 kcal/mol), and chrysin (-7.3 kcal/mol). Cellular experiments validated that these compounds exhibited dose-dependent anti-proliferative effects. Conclusions: The leaves of C. nitidissima induce anti-lung cancer effects via HMOX-1-mediated ferroptosis. Isochlorogenic acid A/C, apigenin, and chrysin were identified as key bioactive components. These findings lay the foundation for the development of natural ferroptosis-targeted drugs.

Keywords: Camellia nitidissima leaves; NSCLC; bioactive screening; ferroptosis; preparative separation.

Abstract

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