Targeting and Inhibiting Plasmodium berghei Growth in Balb/c Mice Using Kojic Acid-Solid Lipid Nanoparticles and Kojic Acid-Nanostructured Lipid Carriers

  • Mehdi Nateghpour* Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Aref Faryabi Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran
  • Afsaneh Motevalli Haghi* Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran
  • Khadijeh Khezri Department of Nursing, Khoy University of Medical Sciences, Khoy, Iran. Neurophysiology Research Center, Cellular and Molecular Medicine Research Institute, Urmia University of Medical Sciences, Urmia, Iran
  • Bahman Rahimi-Esboei Department of Medical Parasitology and Mycology, School of medicine, Islamic Azad University, Tonekabon branch, Iran
  • Abbas Rahimi Foroushani Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Nader Shahrokhi Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran
  • Amir Amani Department of Natural Products and Medicinal Plants Research Center, North Khorasan University of Medical Sciences, Bojnurd, Iran
  • Fatemeh Bayat Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran
Keywords: Malaria, Plasmodium berghei, Kojic acid, Nanoparticles, Nanostructure


  Background and Aim: Malaria is a life-threatening infection in the world. The emergence of strains of Plasmodium species that are resistant to anti-parasitic drugs, and the lack of licensed high-performance malaria vaccines have raised serious concerns worldwide. In recent years, new treatment strategies such as nanoformulations have been suggested as effective drug delivery systems to enhance the therapeutic efficiency of various drugs. Materials and Methods: In this study, kojic acid-solid lipid nanoparticles (KA-SLNs) and kojic acid-nanostructured lipid carriers (KA-NLCs) were synthesized using high-speed homogenization and ultra-probe sonication methods to improve their antiplasmodial activities. The obtained nanoformulations were evaluated against the Plasmodium berghei malaria parasite in mice. Anti-plasmodium activities and cytotoxicity of the nanoparticles were assed. Furthermore, the spleen and liver biochemical analyses of the subjected mice were evaluated for each group of mice in comparison with the control group. Fifty percent effective dose (ED50) was calculated as well. Moreover, ex vivo human red blood cells (RBCs) hemolysis was assessed. Results: Kojic acid solution was significantly effective in all concentrations on the seventh day (D7) and the tenth day (D10) (P. value <0.05). The toxicity test revealed no toxic impact on the subjects. ED50 was obtained at 150 mg/kg concentration for KA-NLCs and 400 mg/kg concentration for KA-SLNs on D10. The results of the evaluation of KA nanoformulations and KA solution on RBCs indicated that KA nanoformulations could reduce the lysis of RBCs.  These results also showed that the lysis of RBCs increased with raising drug concentration in KA nanoformulations, and KA-NLCs (100 mg/kg) gave the least lysis. KA nanoformulations (especially KA-NLCs) and KA solution significantly reduced parasite growth. Conclusion: These results revealed that the KA solution was safe and had no side effects on the subjects in the range of evaluated concentrations. Moreover, the results of this study showed that


World Health Organization(WHO). World malaria report. 2020; Available from: http://

Basir R, Rahiman SF, Hasballah K, Chong W, Talib H, Yam M, et al. Plasmodium berghei ANKA Infection in ICR Mice as a Model of Cerebral Malaria. Iranian Journal of Parasitology. 2012;7(4):62-74.

Druilhe P, Hagan P, Rook GAW. The importance of models of infection in the study of disease resistance. Trends in Microbiology. 2002; 10(10): 38-46.

Olliaro P, Cattani J, Wirth D. Malaria, the submerged disease. Jama. 1996;275(3):230-3.

Haidara M, Haddad M, Denou A, Marti G, Bourgeade Delmas S, Sanogo R, et al. In vivo validation of anti-malarial activity of crude extracts of Terminalia macroptera, a Malian medicinal plant. Malaria Journal. 2018; 17(1): 1-10.

Saeedi M, Eslamifar M, Khezri K. Kojic acid applications in cosmetic and pharmaceutical preparations. Biomedicine & Pharmacotherapy. 2019; 110: 582-93.

Rosita N, Jayanti S, Erawati T. Kojic Acid Penetration: Effect of Carbomer-Tween 80 Agregate Formation. in Proceedings of International Conference on Applied Pharmaceutical Sciences (ICoAPS). 2018.

Khezri K, Saeedi M, Morteza Semnani K, Akbari J, Rostamkalaei SS. An emerging technology in lipid research for targeting hydrophilic drugs to the skin in the treatment of hyperpigmentation disorders: kojic acid-solid lipid nanoparticles. Artificial Cells, Nanomedicine, and Biotechnology, 2020; 48(1): 841-53.

Khezri, K, Saeedi M, Morteza Semnani K, Akbari J, Hedayatizadeh Omran A. A promising and effective platform for delivering hydrophilic depigmenting agents in the treatment of cutaneous hyperpigmentation: kojic acid nanostructured lipid carrier. Artificial Cells, Nanomedicine, and Biotechnology, 2021; 49(1): 38-47.

Omwoyo WN, Ogutu B, Oloo F, Swai H, Kalombo L, Melariri P, et al. Preparation, characterization, and optimization of primaquine-loaded solid lipid nanoparticles. International Journal of Nanomedicine., 2014; 9: 3865-74.

Agbo CP, Ugwuanyi TC, Ugwuoke WI, McConvilla C, Attama AA, Ofokansi KC. Intranasal artesunate-loaded nanostructured lipid carriers: A convenient alternative to parenteral formulations for the treatment of severe and cerebral malaria. Journal of Controlled Release. 2021; 334: 224-36.

Rashidzadeh H, Salimi M, Sadighian S, Rostamizadeh K, Ramazani A. In vivo antiplasmodial activity of curcumin-loaded nanostructured lipid carriers. Current drug delivery. 2019. 16(10): 923-30.

Paliwal R, Paliwal S, Kenwat R, kurmi BD, Sahu MK. Solid lipid nanoparticles: A review on recent perspectives and patents. Expert opinion on therapeutic patents, 2020; 30(3): 179-194.

de Oliveira IF, Barbosa EJ, Peters MCC, Henostroza MAB, Yukuyama MN, Neto EDS, et al. Cutting-edge advances in therapy for the posterior segment of the eye: Solid lipid nanoparticles and nanostructured lipid carriers. International Journal of Pharmaceutics. 2020; 589: 119831.

Araujo VHS, Di Filippo LD, Duarte JL, Sposito L, de Camargo BAF, da Silva PB, et al. Exploiting solid lipid nanoparticles and nanostructured lipid carriers for drug delivery against cutaneous fungal infections. Critical reviews in microbiology. 2021; 47(1): 79-90.

Salvi VR, Pawar P. Nanostructured lipid carriers (NLC) system: A novel drug targeting carrier. Journal of Drug Delivery Science and Technology. 2019; 51: 255-67.

Montazeri M, Emami S, Asgarian-Omran H, Azizi S, Sharif M, Sarvi Sh, et al. In vitro and in vivo evaluation of kojic acid against Toxoplasma gondii in experimental models of acute toxoplasmosis. Experimental Parasitology. 2019; 200: 7-12.

Lenzi J, Costa TM, Alberton MD, Goulart JAG, Tavares LBB. Medicinal fungi: a source of antiparasitic secondary metabolites. Applied microbiology and biotechnology. 2018; 102(14): 5791-810.

Rodrigues APD, Farias LHS, Carvalho ASC, Santos AS, do Nascimento JLM, Silva EO. A novel function for kojic acid, a secondary metabolite from Aspergillus fungi, as antileishmanial agent. PLoS One. 2014; 9(3): e91259.

Liu X, Xu L, Zhan X, Xie W, Yang X, W Cui S, et al. Development and properties of new kojic acid and chitosan composite biodegradable films for active packaging materials. International Journal of Biological Macromolecules. 2020; 144: 483-90.

Fitzpatrick JM, Hiria Y, Hiria H, Hoffmann KF. Schistosome egg production is dependent upon the activities of two developmentally regulated tyrosinases. The FASEB Journal. 2007; 21(3): 823-35.

Peters, W. and B. Robinson, The chemotherapy of rodent malaria. XLVII. Studies on pyronaridine and other Mannich base antimalarials. Annals of Tropical Medicine & Parasitology, 1992; 86(5): 455-65.

Motevalli haghi A, Nateghpour M, Edirssian GhH, Sori E, Satvat MT. Evaluation Of The Effectivenrss Of Ethanolic Extract of Peganum harmala L. Against Plasmodium berghel In Comparison with Chloroquine in sourian mice using in vivo tests. Journal of School of Public Health and Institute of Public Health Research. 2003; 2(1): 47-54.

Karbalaei Pazoki Z, Nateghpour M, Maghsood AH, Souri E, Motevali Haghi A, Farivar L, et al. Comparison between the effects of ethanolic extract of Artemisia annua and chloroquine on Plasmodium berghei in white mice. Scientific Journal of Kurdistan University of Medical Sciences. 2014; 19(2): 9-20.

Khodadadi M, Nateghpour M, Souri E, Motevali Haghi A, Rahimi Froushani A, Karbalaei Z. Evaluation of effectiveness of ethanolic extract of Artemisia aucheri, individually and in combination with chloroquine, on chloroquine-sensitive strain of Plasmodium berghei in sourian mice. Iranian Journal of Public Health. 2013; 42(8): 883-8.

Pestechian N, Abedi Madseh S, Ghanadian M, Nateghpour M. Effect of Crocus sativus Stigma (saffron) alone or in combination with chloroquine on chloroquine sensitive strain of Plasmodium berghei in mice. Journal of HerbMed Pharmacology. 2015. 4(4): 110-14.

Original Article