Identification and Validation of the VEGF/p38MAPK/HSP27 Pro-tumor Inflammatory Pathway: Screening of Active Components from Patrinia Villosa and Evaluation of Their Drug-likeness

Xiaochen Li^1,2,3Yang Ju⁴Xinxin Yang^1,2,3,5Tianjiao Li^1,2,3Shuai Wang^1,2,3,5Yongrui Bao^1,2,3,5*Xiansheng Meng^1,2,3

• ¹School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Da Lian, China
• ²Liaoning Multi-dimensional Analysis of Traditional Chinese Medicine Technical Innovation Center, Dalian, China
• ³Liaoning Province Modern Chinese Medicine Research Engineering Laboratory, Dalian, China
• ⁴Shenyang Orthopaedic Hospital, Shenyang, China
• ⁵Shenyang Key Laboratory for Causes and Drug Discovery of Chronic Diseases, Shenyang, China

Colorectal cancer (CRC) remains one of the leading causes of cancer related mortality worldwide, with pro-tumor inflammation playing a critical role in its initiation and progression. Chronic inflammation not only serves as a major driving force in CRC development but also represents a distinct mechanism of tumorigenesis. Previous studies have demonstrated that the VEGF/p38MAPK and p38MAPK/HSP27 signaling pathways each play important roles in CRC associated inflammation. However, a comprehensive understanding of the entire pro-tumor inflammatory mechanism is still lacking. This mechanism includes membrane level signal perception, cytoplasmic signal transduction, nuclear response, and apoptosis regulation. Our preliminary research revealed that anti-inflammatory and anti-tumor effects are closely associated with signaling pathways such as PI3K and p38MAPK. In this study, we focused on the p38MAPK pathway and, through a combination of network pharmacology and in vivo pharmacodynamic analysis using a p38MAPK pathway inhibitor, systematically identified and validated the pro-tumor inflammatory VEGF/p38MAPK/HSP27 signaling axis. Western blot analysis further confirmed VEGF, p38MAPK, and HSP27 as key target proteins within this pathway. We subsequently identified three active compounds rutin, nicotiflorin, and 4,5-Dicqa from the traditional anti-inflammatory herb Patrinia villosa (PV) through molecular docking and microscale thermophoresis (MST) experiments, based on their high-affinity binding to the aforementioned targets. Finally, molecular dynamics (MD) simulations were employed to evaluate the stability of these compound–target interactions and to assess their potential drug-likeness. Collectively, this study provides both theoretical and experimental evidence for identifying pharmacological targets involved in pro-tumor inflammation. It also offers valuable insights for the development novel therapeutics based on key inflammatory targets.