Immunoadjuvant chemotherapy of visceral leishmaniasis in hamsters using Amphotericin 1

39 The accessible treatment options for life-threatening neglected visceral leishmaniasis (VL) 40 disease have problems regarding efficacy, stability, adverse effects and cost, making treatment a 41 complex issue. Herein, we formulated nanometric amphotericin B (AmB) encapsulated chitosan42 nanocapsules (CNC-AmB) using polymer deposition technique mediated by nano-emulsion 43 template fabrication. CNC-AmB exhibited good steric stability in vitro where chitosan content 44 was found to be efficient in preventing their destabilization in the presence of protein and Ca. 45 Toxicity study on J774A model cell line and erythrocytes revealed less toxicity of CNC-AmB 46 compared to commercialized AmB formulations such as Fungizone® and AmBisome®. 47 Experimental results of in vitro (macrophage amastigote system, IC50 = 0.19±0.04 μg AmB/ml) 48 and in vivo (Leishmania donovani infected hamsters, 86.1±2.08% parasite inhibition) in 49 conjunction with effective internalization by macrophages illustrated the efficacy of CNC-AmB 50 to augment antileishmanial property. Quantitative mRNA analysis by RT-PCR showed that 51 improved effect was synergized with up regulated Tumor Necrosis Factor-α (TNF-α), 52 Interleukin-12 (IL-12) and inducible nitric oxide synthase, and down regulated Transforming 53 Growth Factor-β (TGFβ), IL-10 and IL-4. These research findings suggest that developed cost54 effective CNC-AmB immunoadjuvant chemotherapeutic delivery system could be a viable 55 alternative to the current high-cost commercial lipid based formulations. 56 57