Extraction, Purification and Characterization of Immunomodulatory Effects of Cyclotides Isolated from Viola Odorata

Document Type : Original Article(s)

Authors

1 Department of Pharmaceutics, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Toxicology and Pharmacology, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran.

3 Department of Pharmacognosy, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

4 Department of Chemistry, School of Basic Sciences, Azarbaijan Shahid Madani University, Tabriz, Iran.MSc Student, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Plant cyclic peptides have displayed a diverse range of biological activities. Cyclic peptides exist in different plant families, such as Violaceae.
Methods: In this study, the existence of cyclic peptides in Viola odorata has been investigated. The plant materials were subjected to maceration in methanol: CH2-Cl2 (1:1; v/v), and then the crude extract passed through the C18 column by vacuum liquid chromatography method and fractionated into 50% and 80 % ethanol extract. The obtained fractions were analyzed by HPLC and MALDI-TOF. Finally, the 50% and 80% fractions were injected into the female C57BL/6 mice intraperitoneally at doses of 5, 50, and 100 mg/kg. The lymphocyte, monocyte, neutrophil, eosinophil, and white blood cell (WBC) counts of whole blood in different groups were compared before and 24 hours after the injection. The results confirmed the presence of cyclic peptides in Viola odorata.
Findings: The MALDI-TOF showed the mass weights of fractions were in the range of cyclic peptides. The 80% fraction at all doses reduced the number of lymphocytes and WBC. On the other hand, interestingly, the 50% fraction increased them.
Conclusion: The cyclic peptides can be used as immunomodulatory agents and, thus, can be effective in a wide range of diseases related to the immune system.

Highlights

Ladan Dayani: Google Scholar, PubMed

Azade Taheri: Google Scholar, PubMed

Keywords

Main Subjects

References

  1. Afroz M, Akter S, Ahmed A, Rouf R, Shilpi JA, Tiralongo E, et al. Ethnobotany and antimicrobial peptides from plants of the solanaceae family: an update and future prospects. Front Pharmacol 2020; 11: 565.
  2. Ovesen RG, Göransson U, Hansen SH, Nielsen J, Hansen HCB. A liquid chromatography-electrospray ionization-mass spectrometry method for quantification of cyclotides in plants avoiding sorption during sample preparation. J Chromatogr A 2011; 1218(44): 7964–70.
  3. Del Gatto A, Saviano M, Zaccaro L. An overview of peptide-based molecules as potential drug candidates for multiple sclerosis. Molecules 2021; 26(17): 5227.
  4. Camarero JA, Campbell MJ. The potential of the cyclotide scaffold for drug development. Biomedicines 2019; 7(2): 31.
  5. Narayani M, Chadha A, Srivastava S. Cyclotides from the Indian Medicinal Plant Viola odorata (Banafsha): Identification and Characterization. J Nat Prod 2017; 80(7): 1972–80.
  6. Kwon S, Duarte JN, Li Z, Ling JJ, Cheneval O, Durek T, et al. Targeted Delivery of Cyclotides via Conjugation to a Nanobody. ACS Chem Biol 2018; 13(10): 2973–80.
  7. Nguyen GKT, Zhang S, Nguyen NTK, Nguyen PQT, Chiu MS, Hardjojo A, et al. Discovery and characterization of novel cyclotides originated from chimeric precursors consisting of albumin-1 chain a and cyclotide domains in the Fabaceae family. J Biol Chem 2011; 286(27): 24275–87.
  8. Rosengren KJ, Daly NL, Plan MR, Waine C, Craik DJ. Twists, knots, and rings in proteins. Structural definition of the cyclotide J Biol Chem 2003; 278(10): 8606–16.
  9. Zhang H, Chen S. Cyclic peptide drugs approved in the last two decades (2001–2021). RSC Chem Biol 2022; 3(1): 18-31.
  10. Mourenza A, Ganesan R, Camarero JA. Resistance is futile: targeting multidrug-resistant bacteria with de novo Cys-rich cyclic polypeptides. RSC Chem Biol 2023; 4(10): 722-35.
  11. Huang YH, Jiang Z, Du Q, Yap K, Bigot A, Kaas Q, Wang CK, Craik DJ. Scanning mutagenesis identifies residues that improve the long-term stability and insecticidal activity of cyclotide kalata B1. J Biol Chem 2024; 300(3): 105682.
  12. Azmi S, Mustafa M, Shoaib S, Hussain MK. Structures, functions and therapeutic potential of cyclotides. JERP 2022; 7(4): 234-42.
  13. Conzelmann C, Muratspahić E, Tomašević N, Münch J, Gruber CW. In vitro inhibition of HIV-1 by cyclotide-enriched extracts of Viola tricolor. Front Pharmacol 2022; 13: 888961.
  14. Chia LY, Kumar PV, Maki MA, Ravichandran G, Thilagar S. A review: The antiviral activity of cyclic peptides. Int J Pept Res Ther 2022; 29(1): 7.
  15. Poth AG, Colgrave ML, Lyons RE, Daly NL, Craik DJ. Discovery of an unusual biosynthetic origin for circular proteins in legumes. Proc Natl Acad Sci U S A 2011; 108(25): 10127-32.
  16. Attah A, Fagbemi A, Olubiyi O, Dada-Adegbola H, Oluwadotun A, Elujoba A, et al. Therapeutic Potentials of Antiviral Plants Used in Traditional African Medicine With COVID-19 in Focus: A Nigerian Perspective. Front Pharmacol 2021; 12: 596855.
  17. Herrmann A, Burman R, Mylne JS, Karlsson G, Gullbo J, Craik DJ, et al. The alpine violet, Viola biflora, is a rich source of cyclotides with potent cytotoxicity. Phytochemistry 2008; 69(4): 939–52.
  18. Pränting M, Lööv C, Burman R, Göransson U, Andersson DI. The cyclotide cycloviolacin O2 from Viola odorata has potent bactericidal activity against Gram-negative bacteria. J Antimicrob Chemother 2010; 65(9): 1964–71.
  19. Slazak B, Jacobsson E, Kuta E, Göransson U. Exogenous plant hormones and cyclotide expression in Viola uliginosa (Violaceae). Phytochemistry 2015; 117: 527–36.
  20. Nguyen KNT, Nguyen GKT, Nguyen PQT, Ang KH, Dedon PC, Tam JP. Immunostimulating and Gram-negative-specific antibacterial cyclotides from the butterfly pea (Clitoria ternatea). FEBS J 2016; 283(11): 2067–90.
  21. Bourque J, Hawiger D. Current and future immunotherapies for multiple sclerosis. Mo Med 2021; 118(4): 334-9.
  22. Gründemann C, Koehbach J, Huber R, Gruber C. Do plant cyclotides have potential as immunosuppressant peptides? J Nat Prod 2012; 75: 167–74.
  23. Gründemann C, Stenberg KG, Gruber CW. T20K: An Immunomodulatory Cyclotide on Its Way to the Clinic. Int J Pept Res Ther 2019; 25(1): 9–13.
  24. Fahradpour M, Keov P, Tognola C, Perez-Santamarina E, McCormick PJ, Ghassempour A, et al. Cyclotides isolated from an ipecac root extract antagonize the corticotropin releasing factor type 1 receptor. Front Pharmacol 2017; 8: 616.
  25. Claeson P, Göransson U, Johansson S, Luijendijk T, Bohlin L. Fractionation protocol for the isolation of polypeptides from plant biomass. J Nat Prod 1998; 61(1): 77–81.
  26. Hashempour H, Koehbach J, Daly NL, Ghassempour A, Gruber CW. Characterizing circular peptides in mixtures: Sequence fragment assembly of cyclotides from a violet plant by MALDI-TOF/TOF mass spectrometry. Amino Acids 2013; 44(2): 581–95.
  27. Ji X, Nielsen AL, Heinis C. Cyclic peptides for drug development. Angew Chem Int Ed Engl 2024; 63(3): e202308251.
  28. Park S, Yoo K-O, Marcussen T, Backlund A, Jacobsson E, Rosengren KJ, et al. Cyclotide evolution: insights from the analyses of their precursor sequences, structures and distribution in violets (Viola). Front Plant Sci 2017; 8: 2058.
  29. Poth AG, Colgrave ML, Philip R, Kerenga B, Daly NL, Anderson MA, et al. Discovery of cyclotides in the Fabaceae plant family provides new insights into the cyclization, evolution, and distribution of circular proteins. ACS Chem Biol 2011; 6(4): 345–55.
  30. Fazeenah A, Quamri MA. Banafsha (Viola Odorata Linn.) -A review. World J Pharm Res 2020; 9(10): 514–37.
  31. Slazak B, Kapusta M, Strömstedt AA, Słomka A, Krychowiak M, Shariatgorji M, et al. How Does the Sweet Violet (Viola odorata L.) Fight Pathogens and Pests – Cyclotides as a Comprehensive Plant Host Defense System. Front Plant Sci 2018; 9: 1296.
  32. Svangård E, Göransson U, Hocaoglu Z, Gullbo J, Larsson R, Claeson P, et al. Cytotoxic cyclotides from Viola tricolor. J Nat Prod 2004; 67(2): 144–7.
Journal of Isfahan Medical School
Volume 42, Issue 777
1st Week, October
September and October 2024
Pages 657-665

Files

History

  • Receive Date: 31 August 2024
  • Revise Date: 15 October 2024
  • Accept Date: 27 October 2024

Share

How to cite

Statistics