Inhibitors Targeting Multiple Janus Kinases From Zanthoxylum simulans Mediate Inhibition and Apoptosis Against Gastric Cancer Cells via the Estrogen Pathway

Abstract

Janus kinases (JAKs) play a key role in subtly regulating proliferation, apoptosis, and differentiation of cancer cells, and their inhibitors are actively sought as new drug leads. By developing JAKs based affinity ultrafiltration method coupled with LC/Q-TOF-MS in order to discover selective JAKs inhibitors from total quaternary alkaloids (QAs) from Zanthoxylum simulans, peak 19 (Berberine) and peak 21 (Chelerythrine) were revealed to exhibit notable selectivity on JAK1, JAK2, and JAK3 over Tyk2. In addition, Chelerythrine showed stronger inhibitory activity than the positive control (Cerdulatinib) on gastric cancer cells (AGS), while Berberine, with weaker inhibition. Chelerythrine and Berberine also showed obvious inhibition on human hepatocyte cells (LO2). Furthermore, molecular docking analysis revealed their discrepancies due to different interaction bonds and characteristic residues. Quaternary N was proposed as the functional group to enhance the selectivity of JAK1, and some specific moieties towards Asp1021, Leu855, and Leu828 were suggested to increase the selectivity for JAK1, JAK2, and JAK3, respectively. As the most potential inhibitor of JAKs from QAs, Chelerythrine exhibited distinct suppression of adhesion, migration, invasion, and stimulating apoptosis of AGS cells, which was consistent with the significant down-regulation of estrogen receptors (ER-α36, ER-α66, and ER-β1) and Src expression. In conclusion, an efficient screening approach was developed to identify Berberine and Chelerythrine as potential selective candidates from Zanthoxylum simulans with significant anti-proliferative activity against gastric carcinoma. As we know, it was the first report to propose an estrogen signal pathway for Chelerythrine in anti-gastric cancer cells (AGS) study. The results supported Chelerythrine inhibitory effects on AGS by not only direct inhibiting JAKs but also down-regulating the estrogen pathway.

Molecular docking and molecular dynamics approach to identify EGFR and HER2 receptors inhibitory effect of compounds in Zanthoxylum simulans Hance

“…11 In addition, many studies have demonstrated the anticancer effects of this medicinal herb such as stomach cancer, prostate cancer, leukemia, etc. 12,13 According to the study of Ha Do Thi et al, Z. simulans seed extract showed cytotoxicity to HT-1080 with an IC50 value of 7.1 μg/mL. The leaf extract of Z. simulans also could inhibit many cancer cell lines such as A549, Huh-7, and HT-1080 with IC50 values of 2.7 ± 0.2, 3.2 ± 0.0, 2.8 ± 0.0 μg/mL, respectively.…”

“…This plant has been used in traditional medicine as a medicine mainly used to relieve pain, anti‐inflammation, and anti‐malarial 11 . In addition, many studies have demonstrated the anti‐cancer effects of this medicinal herb such as stomach cancer, prostate cancer, leukemia, etc 12,13 . According to the study of Ha Do Thi et al., Z. simulans seed extract showed cytotoxicity to HT‐1080 with an IC50 value of 7.1 µg/mL.…”

Dual COX-2/5-LOX inhibitors from Zanthoxylum simulans inhibit gastric cancer cells by cross-mediating thyroid, estrogen, and oxytocin signaling pathways

“…Therefore, further studies should be carried out to further explore the bio-active substances and potential mechanisms. Because of multiple substances detected in QAs, it is not easy to figure out the bio-active ingredients and illustrate the potential mechanisms ( Tian et al, 2022 ). Compound isolation combined with activity analysis is a traditional way to discover potential prodrugs, but the time-consuming method may not match the need of improving research efficiency ( Tian et al, 2022 ).…”

“…Because of multiple substances detected in QAs, it is not easy to figure out the bio-active ingredients and illustrate the potential mechanisms ( Tian et al, 2022 ). Compound isolation combined with activity analysis is a traditional way to discover potential prodrugs, but the time-consuming method may not match the need of improving research efficiency ( Tian et al, 2022 ). Ultrafiltration targeting of enzymes has been reported to fish potential bio-active candidates in recent years ( Tian et al, 2022 ).…”

“…Compound isolation combined with activity analysis is a traditional way to discover potential prodrugs, but the time-consuming method may not match the need of improving research efficiency ( Tian et al, 2022 ). Ultrafiltration targeting of enzymes has been reported to fish potential bio-active candidates in recent years ( Tian et al, 2022 ). For the exploration of mechanisms, some bio-informatics methods, including Gene Expression Omnibus (GEO), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG), are helpful to indicate possible mechanism assays based on public and massive data, which are reported to find the genes with significantly different expression, enrich possible pathways, and clarify the most valuable regulations ( Shi et al, 2022 ; Bing et al, 2023 ; Quan et al, 2023 ).…”