Veterinary Research Forum
Vet Res Forum

In vitro evaluation of activatable melittin encapsulated in liposome and albumin nanoparticles against Leishmania

Document Type : Original Article

Authors

Soheila Akhzari 1
Sedigheh Nabian 2
Mohammad Taheri 3

1 Department of Animal Science, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran

2 Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Rastegar Reference Laboratory, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

https://doi.org/10.30466/vrf.2025.2047654.4572
Abstract
Leishmaniasis comprises a spectrum of clinical manifestations caused by protozoan parasites of the genus Leishmania, order Trypanosomatida. Cutaneous leishmaniasis remains a significant zoonotic disease prevalent in tropical and subtropical regions, particularly in developing countries. Despite ongoing research, a definitive cure for this parasitic infection is still needed. This study explored the potential of activatable melittin (AM) as a selective treatment for cutaneous leishmaniasis caused by Leishmania major. The AM was designed using PepFold and ExPASy servers, incorporating a matrix metalloproteinase -2/9 cleavable linker to target L. major-infected macrophages selectively. To enhance drug delivery and reduce potential toxicity, AM was encapsulated within albumin nanoparticles and liposomes. The anti-leishmanial efficacy of these formulations was evaluated at AM concentrations ranging from 25.00 to 100 µg mL-1 over 48 hr, with each experiment performed in 10 independent replicates (n = 10 per group). Statistical analysis using one-way ANOVA followed by Tukey's post-hoc test revealed a significant reduction in the average number of intracellular amastigotes per macrophage in the liposome-treated and albumin nanoparticle-treated groups (7.00 ± 1.50 amastigotes per macrophage) compared to the untreated infected control group (35.00 ± 3.20 amastigotes per macrophage). Treatment with 25.00 µg mL-1 of AM encapsulated in non-toxic albumin nanoparticles and liposomes demonstrated the most promising anti-leishmanial effect, resulting in an approximately 80.00% reduction in intracellular L. major amastigotes (compared to control).

Keywords

Cytotoxicity
Drug delivery
Leishmania major
Melittin
Subjects
1.     Akhzari S, Nabian S, Taheri M, et al. Therapeutic evaluation of activatable melittin for selective treatment of leishmaniasis in vitro and in vivo. Bulg J Vet Med 2023; 27(1): 0019. doi: 10.15547/ bjvm.2022-0019.
2.     Akhzari S, Rezvan H, Zolhavarieh SM, et al. Comparison of proinflammatory gene expression in lesions caused by either burn injuries or cutaneous leishmaniasis. Gene Cell Tissue 2017; 4(1): e42887. doi: 10.17795/gct-42887.
3.     Akhzari S, Rezvan H, Nourian A, et al. Evaluation of pro-inflammatory genes expression in the spleen and wounds of BALB/c mice infected with Leishmania major [Persian]. J Ilam Univ Med Sci 2018; 26(2): 77-86.
4.     Razavi MR, Shirzadi MR, Mohebali M, et al. Human cutaneous leishmaniasis in Iran, up to date-2019. J Arthropod Borne Dis 2021; 15(2): 143-151.
5.     Rezvan H, Hamoon Navard S, Akhzari S. Comparison of pro-inflammatory gene expression profile in spleen and peripheral blood neutrophils of BALB/C mice infected with Leishmania major [Persian]. JSSU 2022; 29(11): 4268-4280.
6.     Salari S, Bamorovat M, Sharifi I, et al. Global distribution of treatment resistance gene markers for leishmaniasis. J Clin Lab Anal 2022; 36(8): e24599. doi: 10.1002/jcla.24599.
7.     Sangenito LS, da Silva Santos V, d'Avila-Levy CM, et al. Leishmaniasis and Chagas disease - neglected tropical diseases: treatment updates. Curr Top Med Chem 2019; 19(3): 174-177.
8.     Tichy J, Novak J. Detection of antimicrobials in bee products with activity against viridans streptococci. J Altern Complement Med 2000; 6(5): 383-389.
9.     Wade D, Boman A, Wåhlin B, et al. All-D amino acid-containing channel-forming antibiotic peptides. Proc Natl Acad Sci U S A 1990; 87(12): 4761-4765.
10. Bazzo R, Tappin MJ, Pastore A, et al. The structure of melittin. A 1H‐NMR study in methanol. Eur J Biochem 1988; 173(1): 139-146.
11. Adade CM, Oliveira IR, Pais JA, et al. Melittin peptide kills Trypanosoma cruzi parasites by inducing different cell death pathways. Toxicon 2013; 69: 227-239.
12. Olson ES, Aguilera TA, Jiang T, et al. In vivo characterization of activatable cell penetrating peptides for targeting protease activity in cancer. Integr Biol (Camb) 2009; 1(5-6): 382-393.
13. Kulkarni MM, McMaster WR, Kamysz W, et al. Antimicrobial peptide-induced apoptotic death of Leishmania results from calcium-dependent, caspase-independent mitochondrial toxicity. J Biol Chem 2009; 284(23): 15496-15504.
14. Alberola J, Rodríguez A, Francino O, et al. Safety and efficacy of antimicrobial peptides against naturally acquired leishmaniasis. Antimicrob Agents Chemother 2004; 48(2): 641-643.
15. Zarrinnahad H, Mahmoodzadeh A, Hamidi MP, et al. Apoptotic effect of melittin purified from Iranian honey bee venom on human cervical cancer HeLa cell line. Int J Pept Res Ther 2018; 24(4): 563-570.
16. Akhzari S, Nabian S, Shayan P, et al. Designing of RNA molecule translating for activitable melittin as selective targeting of Leishmania infected cells. Iran J Parasitol 2021; 16(3): 443-453.
17. Tempone AG, da Silva AC, Brandt CA, et al. Synthesis and antileishmanial activities of novel 3-substituted quinolines. Antimicrob Agents Chemother 2005; 49(3): 1076-1080.
18. Ghanbarzadeh S, Valizadeh H, Zakeri-Milani P. Application of response surface methodology in development of sirolimus liposomes prepared by thin film hydration technique. Bioimpacts 2013; 3(2): 75-81.
19. Pacheco-Fernandez T, Markle H, Verma C, et al. Field-deployable treatments for leishmaniasis: intrinsic challenges, recent developments and next steps. Res Rep Trop Med 2023; 14: 61-85.
20. Vartak DG, Gemeinhart RA. Matrix metalloproteases: underutilized targets for drug delivery. J Drug Target 2007; 15(1): 1-20.
21. Moghimipour E, Aghel N, Zarei Mahmoudabadi A, et al. Preparation and characterization of liposomes containing essential oil of Eucalyptus camaldulensis leaf. Jundishapur J Nat Pharm Prod 2012; 7(3): 117-122.
22. Mahmoodzadeh A, Zarrinnahad H, Bagheri KP, et al. First report on the isolation of melittin from Iranian honey bee venom and evaluation of its toxicity on gastric cancer AGS cells. J Chin Med Assoc 2015; 78(10): 574-583.
23. Vuarchey C, Kumar S, Schwendener RA. Albumin coated liposomes: a novel platform for macrophage specific drug delivery. Nanotechnol Devt 2011; 1(1): e2. doi: 10.4081/nd.2011.e2
24. Veerareddy PR, Vobalaboina V, Nahid A. Formulation and evaluation of oil-in-water emulsions of piperine in visceral leishmaniasis. Pharmazie 2004; 59(3): 194-197.
25. Su CW, Chiang CS, Li WM, et al. Multifunctional nanocarriers for simultaneous encapsulation of hydrophobic and hydrophilic drugs in cancer treatment. Nanomedicine (Lond) 2014; 9(10): 1499-1515.
26. Cui Z, Zhou Z, Sun Z, et al. Melittin and phospholipase A2: promising anti-cancer candidates from bee venom. Biomed Pharmacother 2024; 179: 117385. doi: 10.1016/j.biopha.2024.117385.
27. Malam Y, Loizidou M, Seifalian AM. Liposomes and nanoparticles: nanosized vehicles for drug delivery in cancer. Trends Pharmacol Sci 2009; 30(11): 592-599.
28. Gour JK, Srivastava A, Kumar V, et al. Nanomedicine and leishmaniasis: future prospects. Dig J Nanomater Biostructures 2009; 4(3): 495-499.
Veterinary Research Forum
Volume 17, Issue 2
February 2026
Pages 127-133

  • Receive Date 28 December 2024
  • Revise Date 12 May 2025
  • Accept Date 14 July 2025

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