Potential antitumor activity of a new plant-derived alkaloid isolated from Petasites hybridus (L.), in silico and in vitro

Introduction. Some cytostatic drugs are based on the compounds of plant origin. Development of the new plant metabolites-derived antitumor drugs is an actual and perspective trend in oncology. Recently we isolated a compound from the rhizome of Petasites hybridus (L.) and identified it as a corynan-like indole alkaloid P1.

Aim. To identify of the possibility of the Petasites hybridus (L.)-derived alkaloid to bind with molecular targets mediating carcinogenesis and tumor growth and the assessment of its effect on tumor and normal cell cultures.

Materials and methods. The research was performed in silico by molecular docking and in vitro by cultural methods. Molecular docking of alkaloid Р1 with receptors of epidermal growth factor (EGFR), platelet growth factor (PDGFR), MET, MRP2 and NOX4 was carried out using software AutoDock Vina 4.0. The docking area grid was built in AutoDockTools 1.5.7. Cultural experiments were performed on tumor cell line highly expressing EGFR (Н1299) and normal lung fibroblasts. Cells were cultured with various concentrations of the alkaloid, their viability was assessed in ХТТ-test and by direct count of alive and dead cells.

Results. Maximal binding energy of alkaloid Р1 with molecular targets was noted for МеТ (–8,6 kcal/M), minimal – for NOX4 (–5,9 kcal/M). Binding energy of alkaloid Р1 with EGFR was –6,4; with PDGFR –7,2; with MRP2 –6,3 kcal/M. Binding of alkaloid P1 occurred with the amino acid residues of the active centers of the majority of the studied receptors. Alkaloid showed the ability to inhibit the growth of H1299 cell line (lung adenocarcinoma) in the wide range of concentrations more actively than of normal fibroblasts: half-maximal inhibitory concentration for P1 was 127,24 and 256,29 μM/l respectively. Maximal difference of the dead cells amount between H1299 line and fibroblasts was observed in alkaloid concentrations 21,7–87 μM/l, but, in spite of some signs of mitotic failure (multinucleate and giantnucleate H1299 cells) mitotic index did not change.

Conclusion. Alkaloid isolated from the rhizome of Petasites hybridus (L.) is able to bind with the targets mediating tumor growth and impairs lung adenocarcinoma Н1299 cells. Further research is necessary for detailed study of its` antitumor effect in in vitro and in vivo models.

1. Mushtaq S., Abbasi B.H., Uzairm B., Abbasi R. Natural products as reservoirs of novel therapeutic agents. EXCLI J 2018;4(17):420. DOI: 10.17179/excli2018-1174

2. Kit O.I., Zhukova G.V., Tolkachev O.N. et al. Antitumor factors of natural origin and some approaches to the development of effective regimens of phytotherapy in oncology (literature review including the findings of authors’ own research). Voprosy onkologii = Problems in Oncology 2022;68(5):27–38. (In Russ.). DOI: 10.37469/0507-3758-2022-68-5-527-538

3. Morgillo F., Della Corte C.M., Fasano M., Ciardiello F. Mechanisms of resistance to EGFR-targeted drugs: lung cancer. ESMO Open 2016;1(3):e000060. DOI: 10.1136/esmoopen-2016-000060

4. Du H., Gu J., Peng Q. et al. Berberine suppresses EMT in liver and gastric carcinoma cells through combination with TGFβR regulating TGF-β/Smad pathway. Oxid Med Cell Longev 2021;18(21):2337818. DOI: 10.1155/2021/2337818

5. Cosme P., Rodríguez A.B., Espino J., Garrido M. Plant phenolics: bioavailability as a key determinant of their potential health promoting applications. Antioxidants (Basel) 2020;9(12):1263. DOI: 10.3390/antiox9121263

6. Zlatnik E.Yu., Enin Ya.S., Burov O.N. et al. Molecular and cellular aspects of the impact of secondary metabolites from common barberry and hybrid goat’s rue on the HeLa cell line. Issledovaniya i praktika v meditsine = Research’n Practical Medicine Journal 2023;10(4):31 47. (In Russ.). DOI: 10.17709/2410-1893-2023-10-4-3

7. Fu W., Liu L., Tong S. Berberine inhibits the progression of breast cancer by regulating METTL3-mediated m6A modification of FGF7 mRNA. Thorac Cancer 2024;15(17):1357. DOI: 10.1111/1759-7714.15321

8. Achi I.T., Sarbadhikary P., Blassan P.G., Abrahamse H. Multi-target potential of berberine as an antineoplastic and antimetastatic agent: a special focus on lung cancer treatment. Cells 2022;11(21):3433. DOI: 10.3390/cells11213433

9. Hassan S.M., Farid A., Panda S.S. et al. Indole compounds in oncology: therapeutic potential and mechanistic insights. Pharmaceuticals 2024;17(7):922. DOI: 10.3390/ph17070922

10. Roehm N.W., Rodgers G.H., Hatfield S.M., Glasebrook A.L. An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J Immunol Methods 1991;142(2):257–65. DOI: 10.1016/0022-1759(91)90114-u

11. Jabbarzadeh K.P., Leong M.P., Ismail P., Ling K.H. Antitumor effects of berberine against EGFR, ERK1/2, P38 and AKT in MDA-MB231 and MCF-7 breast cancer cells using molecular modelling and in vitro study. Pharmacol Rep 2019;71(1):13. DOI: 10.1016/j.pharep.2018.07.005

12. Huang B., Wen G., Li R. et al. Integrated network pharmacology, bioinformatics, and molecular docking to explore the mechanisms of berberine regulating autophagy in breast cancer. Medicine (Baltimore) 2023;102(36):e35070. DOI: 10.1097/MD.0000000000035070

13. Zhang M., Wang C.M., Li J. et al. Berberine protects against palmitate-induced endothelial dysfunction: involvements of upregulation of AMPK and eNOS and downregulation of NOX4. Mediators Inflamm 2013;(4):260464. DOI: 10.1155/2013/260464

14. Sun A., Yang H., Li T. et al. Molecular mechanisms, targets and clinical potential of berberine in regulating metabolism: a review focussing on databases and molecular docking studies. Front Pharmacol 2024;15(6). DOI: 10.3389/fphar.2024.1368950

15. Hu M., Yan H., Li H. et al. Use of network pharmacology and molecular docking to explore the mechanism of action of curcuma in the treatment of osteosarcoma. Sci Rep 2023;13:9569. DOI: 10.1038/s41598-023-36687-z

16. Kang H.N., Kim S.H., Yun M.R. et al. ER2, a novel human anti-EGFR monoclonal antibody inhibit tumor activity in non-small cell lung cancer models. Lung Cancer 2016;5(95):57–64. DOI: 10.1016/j.lungcan.2016.02.013

17. Liu W.J., Wang L., Zhou F.M. et al. Elevated NOX4 promotes tumorigenesis and acquired EGFR-TKIs resistance via enhancing IL-8/PD-L1 signaling in NSCLC. Drug Resist Updat 2023;9(70):100987. DOI: 10.1016/j.drup.2023.100987

18. Pandey P., Khan F., Upadhyay T.K. et al. New insights about the PDGF/PDGFR signaling pathway as a promising target to develop cancer therapeutic strategies. Biomed Pharmacother 2023;161:114491. DOI: 10.1016/j.biopha.2023.114491

19. Chen Z., Vallega K.A., Chen H. et al. The natural product berberine synergizes with osimertinib preferentially against MET-amplified osimertinib-resistant lung cancer via direct MET inhibition. Pharmacol Res 2022;1(175):105998. DOI: 10.1016/j.phrs.2021.105998

20. Chin T.M., Boopathy G.T.K., Man E.P.S. et al. Targeting microtubules sensitizes drug resistant lung cancer cells to lysosomal pathway inhibitors. Theranostics 2020;10(6):2727–43. DOI: 10.7150/thno.38729