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. 2024 Jul 22:15:1417372.
doi: 10.3389/fphar.2024.1417372. eCollection 2024.

Synthesis, characterization, and mechanistic insights into the enhanced anti-inflammatory activity of baicalin butyl ester via the PI3K-AKT pathway

Hongxu Du  1   2 , Zhangxun Li  1 , Lijuan Su  1 , Zhengke He  1 , Xiaoyan Tan  1 , Fengzhi Hou  1 , Tanjie He  1 , Yu Pan  1 , Shuang Xu  1 , Liting Cao  1   2 , Shiqi Dong  1 , Yue Ma  1   2
Affiliations

Synthesis, characterization, and mechanistic insights into the enhanced anti-inflammatory activity of baicalin butyl ester via the PI3K-AKT pathway

Hongxu Du et al. Front Pharmacol. .

Abstract

Objective: To investigate the anti-inflammatory activity and mechanism of Baicalin derivative (Baicalin butyl ester, BE).

Methods: BE was synthesized and identified using UV-Vis spectroscopy, FT-IR spectroscopy, mass spectrometry (MS) and high-performance liquid chromatography (HPLC) methods. Its anti-inflammatory potential was explored by an in vitro inflammation model. Network pharmacology was employed to predict the anti-inflammatory targets of BE, construct protein-protein interaction (PPI) networks, and analysis topological features and KEGG pathway enrichment. Additionally, molecular docking was conducted to evaluate the binding affinity between BE and its core targets. qRT-PCR analysis was conducted to validate the network pharmacology results. The organizational efficiency was further evaluated through octanol-water partition coefficient and transmembrane activity analysis.

Results: UV-Vis, FT-IR, MS, and HPLC analyses confirmed the successfully synthesis of BE with a high purity of 93.75%. In vitro anti-inflammatory research showed that BE could more effectively suppress the expression of NO, COX-2, IL-6, IL-1β, and iNOS. Network pharmacology and in vitro experiments validated that BE's anti-inflammatory effects was mediated through the suppression of SRC, HSP90AA1, PIK3CA, JAK2, AKT1, and NF-κB via PI3K-AKT pathway. Molecular docking results revealed that the binding affinities of BA to the core targets were lower than those of BE. The Log p-value of BE (1.7) was markedly higher than that of BA (-0.5). Furthermore, BE accumulated in cells at a level approximately 200 times greater than BA.

Conclusion: BE exhibits stronger anti-inflammatory activity relative to BA, possibly attributed to its better lipid solubility and cellular penetration capabilities. The anti-inflammatory mechanism of BE may be mediated through the PI3K-AKT pathway.

Keywords: baicalin butyl ester; inflammation; molecular docking; network pharmacology; traditional Chinese medicine.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Synthesis of BE.
FIGURE 2
FIGURE 2
The characterization of BE. (A) Apparent property of BA and BE. (B) Full wavelength scanning of BA and BE. (C) FT-IR spectrum analysis of BA and BE. (D) Mass spectrometry detection of BE. (E) HPLC chromatograms of BA and BE.
FIGURE 3
FIGURE 3
Anti-inflammatory effect of BA and BE in RAW264.7 cells. (A) Cell viability. (B) NO. (C) IL-1β. (D) IL-6. (E) COX-2. (F) iNOS. Note: #p < 0.05, ##p < 0.01 compared with the CC group; △ p < 0.05, △△ p < 0.01 compared with the LPS group; *p < 0.05, **p < 0.01 compared with BA group (N = 3).
FIGURE 4
FIGURE 4
Network Pharmacology analysis. (A) BA intersects with inflammatory targets. (B) BE intersects with inflammatory targets. (C) Intersect targets PPI diagram of BA. (D) Intersect targets PPI diagram of BE. (E) Topology analysis of BA’s PPI network. (F) Topology analysis of BE’s PPI network. (G) The KEGG signaling pathways of BA. (H) The KEGG signaling pathways of BE.
FIGURE 5
FIGURE 5
Molecular docking analysis.
FIGURE 6
FIGURE 6
Molecular docking detail. Note: Green is the drug structure of BA or BE, blue is the protein, red is the amino acid residues interacting with the drug, and the below right corner of the figure shows the binding position of the drug to the protein.
FIGURE 7
FIGURE 7
Relative mRNA expression level detection. (A) SRC. (B) JAK2. (C) HSP90AA1. (D) PIK3CA. (E) AKT-1. (F) NF-κB. Note: # P < 0.05, ## P < 0.01 compared with the CC group; ▵ P < 0.05,▵▵ P < 0.01 compared with the LPS group; * P < 0.05, ** P < 0.01 compared with BA group (N=3).
FIGURE 8
FIGURE 8
Oil-water partition and transmembrane ability detection of BA and BE. (A) Solubility of BA and BE in octanol and water. (B) log p-value of BA and BE. (C) Transmembrane ability detection of BA and BE.
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