In Vitro Antimalarial Activity of Chloroquine-Crocus Sativus Conjugated to Chitosan Nanocomposits against 3D7 and K1 Strains of Plasmodium falciparum
-
Aram Khezri
,
Mahdi Nateghpour
,
Afsaneh Motevali Haghi
,
Taher Elmi
,
Abbas Rahimi Foroushani
,
Mehdi Shafii Ardestani
,
Haleh Hanifian
Abstract
Background: The use of nanocarriers in combination with other treatments shows significant promise in addressing drug-resistant diseases, particularly malaria. Given the high prevalence of drug-resistant malaria, research into innovative therapies is crucial. This study focuses on a nanoform of chitosan, a biodegradable polymer, combined with Crocus sativus (saffron) and chloroquine to enhance their antimalarial effects.
Methods: Saffron extract and chloroquine were separately conjugated with chitosan, followed by confirmation tests to determine conjugation efficiency. Both chloroquine-resistant and sensitive strains of Plasmodium falciparum were cultured to calculate the IC50 values of various treatments in vitro. This study was conducted at the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2024.
Results: Confirmation tests (FTIR, DLS, Zeta potential, TEM) verified proper drug conjugation to nanocomposites, with observed nanosize, the percentage of conjugation was 64.4% for chloroquine and 42.9% for saffron. Toxicity and hemolysis tests confirmed safe doses. The IC50s values for Chloroquine, Nanoparticle-Chloroquine, Saffron, and Nanoparticle-Saffron were 0.3, 0.8, 42.5, and 6.24 µg/ml, respectively, for the sensitive strain, and 5, 1, 12.5, and 3.12 µg/ml, respectively, for the resistant strain. Combination therapy with the fixed ratio method showed synergistic effects. Statistical analysis revealed synthesized nanocomposites' superior inhibition of P. falciparum growth compared to non-nano. Significant differences were observed in some cases (P< 0.05).
Conclusion: Utilizing nanocarriers and combination therapy is an appropriate strategy for addressing drug resistance. Saffron's anti-malarial effects on P. falciparum were notably increased when linked to chitosan nanocomposites. Furthermore, employing a fixed ratio technique enhanced the therapeutic effectiveness of saffron when combined with chloroquine and chloroquine-nanocomposites across all concentrations.
2. world-malaria-report-2024. Available from: https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2024
3. Gallup JL, Sachs JD. The economic burden of malaria. CID Working Paper No 52. 2000;
4. White N, Pukrittayakamee S, Hien T, Faiz M, Mokuolu O, Dondorp A. Erratum: Malaria Lancet. 2014; 383(9918):723-35.
5. Mita T, Jombart T. Patterns and dynamics of genetic diversity in P. falciparum: what past human migrations tell us about malaria. Parasitol Int. 2015;64(3):238–43.
6. Ataba E, Dorkenoo AM, Nguepou CT, et al. Potential emergence of Plasmodium resistance to artemisinin induced by the use of Artemisia annua for malaria and COVID-19 prevention in Sub-African Region. Acta Parasitol. 2022; 67(1):55-60.
7. Heidari A, Keshavarz H. The drug resistance of Plasmodium falciparum and P. vivax in Iran: a review article. Iran J Parasitol. 2021;16(2):173-185.
8. Million E, Mulugeta T, Umeta B. Traditional medicine practice and its role in the management of malaria in jimma town, Oromia, Ethiopia. Infect Drug Resist. 2022; 2022:2187–98.
9. Appiah EO, Appiah S, Oti-Boadi E, et al. Practices of herbal management of malaria among trading mothers in Shai Osudoku District, Accra. PLoS One. 2022; 17(7):e0271669.
10. Olasehinde GI, Ojurongbe O, Adeyeba AO, et al. In vitro studies on the sensitivity pattern of Plasmodium falciparum to anti-malarial drugs and local herbal extracts. Malar J. 2014; 13:63.
11. Agarwal N, Kolba N, Jung Y, Cheng J, Tako E. Saffron (Crocus sativus L.) flower water extract disrupts the cecal microbiome, brush border membrane functionality, and morphology in vivo (gallus gallus). Nutrients. 2022;14(1):220.
12. Samarghandian S, Borji A. Anticarcinogenic effect of saffron (Crocus sativus L.) and its ingredients. Pharmacognosy Res. 2014;6(2):99-107.
13. Khorasany AR, Hosseinzadeh H. Therapeutic effects of saffron (Crocus sativus L.) in digestive disorders: a review. Iran J Basic Med Sci. 2016;19(5):455-69.
14. Li B, Elango J, Wu W. Recent advancement of molecular structure and biomaterial function of chitosan from marine organisms for pharmaceutical and nutraceutical application. Appl Sci. 2020;10(14):4719.
15. Rashedi J, Haghjo AG, Abbasi MM, et al. Anti-tumor effect of quercetin loaded chitosan nanoparticles on induced colon cancer in wistar rats. Adv Pharm Bull. 2019;9(3):409-415.
16. M. Ways TM, Lau WM, Khutoryanskiy VV. Chitosan and its derivatives for application in mucoadhesive drug delivery systems. Polymers (Basel). 2018;10(3):267.
17. Baharlouei P, Rahman A. Chitin and chitosan: Prospective biomedical applications in drug delivery, cancer treatment, and wound healing. Mar Drugs. 2022;20(7):460.
18. Li J, Cai C, Li J, et al. Chitosan-based nanomaterials for drug delivery. Molecules. 2018;23(10):2661.
19. Kremsner PG, Krishna S. Antimalarial combinations. Lancet. 2004;364(9430):285–94.
20. Alven S, Aderibigbe B. Combination therapy strategies for the treatment of malaria. Molecules. 2019;24(19):3601.
21. Global Partnership to Roll Back Malaria. (2001). Antimalarial drug combination therapy: report of a WHO technical consultation, 4-5 April 2001. World Health Organization.
22. Elmi T, Rahimi Esboei B, Sadeghi F, et al. In vitro antiprotozoal effects of nano-chitosan on Plasmodium falciparum, Giardia lamblia and Trichomonas vaginalis. Acta Parasitol. 2021; 66:39–52.
23. Kamiloglu S, Sari G, Ozdal T, Capanoglu E. Guidelines for cell viability assays. Food Front. 2020;1(3):332–49.
24. Hajialiani F, Elmi T, Mohamadi M, et al. Analysis of the active fraction of Iranian Naja naja oxiana snake venom on the metabolite profiles of the malaria parasite by 1HNMR in vitro. Iran J Basic Med Sci. 2020;23(4):534-543.
25. Mainardes RM, Evangelista RC. PLGA nanoparticles containing praziquantel: effect of formulation variables on size distribution. Int J Pharm. 2005;290(1–2):137–44.
26. Bhattacharyya N, Mukherjee H, Bose D, et al. Clinical case of artesunate resistant Plasmodium falciparum malaria in Kolkata: A first report. J Trop Dis. 2014;2(128):2.
27. Roux AT, Maharaj L, Oyegoke O, et al. Chloroquine and sulfadoxine–pyrimethamine resistance in Sub-Saharan Africa—A review. Front Genet. 2021; 12:668574.
28. Retno S, Widyawaruyanti A, Anindita FBT, Astuti SK, Setyawan D. Development of andrographolide-carboxymethyl chitosan nanoparticles: characterization, in vitro release and in vivo antimalarial activity study. Turk J Pharm Sci. 2018;15(2):136-141.
29. Momenfam F, Nateghpour M, Haghi AM, et al. Interaction between chitosan and chloroquine against Plasmodium berghei and P. falciparum using in-vivo and in-vitro tests. Iran J Parasitol. 2021; 16(2):261-269.
30. Zein U, Fitri LE, Saragih A. Comparative study of antimalarial effect of sambiloto (Andrographis paniculata) extract, chloroquine and artemisinin and their combination against Plasmodium falciparum in-vitro. Acta Med Indones. 2013;45(1):38-43.
31. Popović-Djordjević JB, Kostić AŽ, Kiralan M. Antioxidant activities of bioactive compounds and various extracts obtained from saffron. In: Saffron. Elsevier; 2021. p. 41–97.
32. Azghandi Fardaghi A, Es-haghi A, Feizy J, Lakshmipathy R. Antioxidant capacity and chemical composition of different parts of saffron flowers. Journal of Food and Bioprocess Engineering. 2021;4(1):69–74.
33. Sacco P, Pedroso-Santana S, Kumar Y, Joly N, Martin P, Bocchetta P. Ionotropic gelation of chitosan flat structures and potential applications. Molecules. 2021;26(3):660.
34. Patel B, Tripathi Sk, Shukla S, Pandey A. Preparation, characterization and in vitro drug release study of chloroquine phosphate-loaded chitosan nanoparticles as malaria treatment. Oxid Commun. 2021;44(4).
| Files | ||
| Issue | Vol 20 No 2 (2025) | |
| Section | Original Article(s) | |
| Keywords | ||
| Plasmodium falciparum Saffron Chitosan Therapeutic combination In vitro Nanocomposite | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
