Comparative analysis between four model nanoformulations of amphotericin B-chitosan, amphotericin B-dendrimer, betulinic acid-chitosan and betulinic acid-dendrimer for treatment of Leishmania major: real-time PCR assay plus

Abstract

Tahereh Zadeh Mehrizi,1,2 Ali Khamesipour,3,* Mehdi Shafiee Ardestani,4 Hasan Ebrahimi Shahmabadi,5 Mostafa Haji Molla Hoseini,6 Nariman Mosaffa,6 Amitis Ramezani1,* 1Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran; 2Pishtaz Teb Zaman Diagnostics, Tehran, Iran; 3Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran; 4Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; 5Department of Microbiology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran; 6Department of Medical Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran*These authors contributed equally to this workCorrespondence: Ali KhamesipourCenter for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, 415 Taleghani Avenue, P.O. Box 14155-6383, Tehran 1416613675, IranTel [phone]Email [email protected] RamezaniClinical Research Dept, Pasteur Institute of Iran, 13164, Pasteur Ave., Tehran 1416613675, IranTel [phone]Email [email protected]: Amphotericin B (Amp) and Betulinic acid (BA) as antileishmanial agents have negligible water solubility and high toxicity. To solve these problems, for the first time, chitosan nanoparticles and Anionic Linear Globular Dendrimer (D) were synthesized for the treatment of Leishmania major (L. major).Method: Chitosan and dendrimer nanoparticles were synthesized, and Amp and BA were loaded into the nanoparticles. The particles were then characterized using various methods and their efficacy was evaluated in vitro and in vivo environments (parasite burden was confirmed using pathological studies and real-time PCR methods).Result: The results of docking showed that Amp and BA can be loaded into chitosan and dendrimer nanoparticles. The results of physically drug loading efficiency for AK (Amphotericin B-chitosan), BK (Betulinic acid-chitosan), AD (Amphotericin B-Dendrimer) and BD (Betulinic acid- Dendrimer) were 90, 93, 84 and 96 percent, respectively. The characterization results indicated that the drugs were loaded into nanoparticles physically. Moreover, the increased solubility rate for AD=478, BD=790, AK=80 and BK=300 folds. Furthermore, the results of the drug delivery system showed the slow controlled drug release pattern with cellular uptake of more than 90%. The treatment results showed a 100 percent decrease of toxicity for the all nanodrugs was observed in vivo and in vitro environments. Moreover, AK10 and BK20 mg/kg reduced parasite burden by 83 percent (P<0.001), while AD50 and BD40 mg/kg reduced it to a lesser extent compared to glucantime.Conclusion: All the synthesized nanodrugs were completely succeeded by 100% to recovery the L. major induced pathological effects in the infected footpad. Also, the results of present study were confirmed with real-time PCR and the results showed that AK and BK were succeeded in a large extent to the treatment of L. major infection (P<0.001), therefore AK and BK could be considered as proper alternatives of choices drugs.Keywords: chitosan, L. major, anionic linear globular dendrimer, treatment

Keywords

• chitosan
• L. Major
• Anionic Linear Globular Dendrimer
• treatment.