Enhanced Topical Treatment of Cutaneous Leishmaniasis Using Niosomal Gel of Tioxolone-Benzoxonium Chloride: In silico, in vitro, and in vivo Evaluation

Maryam Hakimi Parizi,1 Iraj Sharifi,1 Elaheh Molaakbari,1 Ehsan Salarkia,1 Mahshid Mostafavi,1 Mohammad Hossein Daie Parizi,1 Abbas Pardakhty,2 Ali Khamesipour,3 Daryoush Ghaffari11Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran; 2Pharmaceutics Research Center, Pharmaceutical Sciences Institute, Kerman University of Medical Sciences, Kerman, Iran; 3Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, IranCorrespondence: Iraj Sharifi, Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, 76169-14115, Iran, Tel +98-341-3224616, Fax +98-341-3239843, Email iraj.sharifi@yahoo.com Elaheh Molaakbari, Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, 76169-14115, Iran, Tel +98-341-3224616, Fax +98-341-3239843, Email e.molaakbari@gmail.comBackground: Cutaneous leishmaniasis (CL) is a neglected tropical disease that poses significant therapeutic challenges owing to its toxicity and low compliance with existing treatments. In this study, we formulated and evaluated niosomal gels containing tioxolone (NT), benzoxonium chloride (NB), and their combination (NTB) for topical application against CL.Methods: Various in silico methods, including homology modeling and molecular docking, were used to assess pharmacological potential. In vitro anti-leishmanial activity was tested against Leishmania major promastigotes and amastigotes, whereas in vivo efficacy was evaluated in a BALB/c mouse model.Results: Docking studies showed strong interactions between NT, NB, and NTB and the target proteins. The ADMET analysis indicated that NTB had favorable aqueous solubility (LogS = − 3.71) and low toxicity (2.88 mol/kg). In vitro, NTB demonstrated the lowest IC50 values (79.0± 3.2 μg/mL for promastigotes and 44.4± 0.8 μg/mL for amastigotes) against both parasite forms and the highest selectivity index (9.8), outperforming the standard drug meglumine antimoniate (MA). In vivo tests revealed that NTB significantly reduced lesion size and splenic parasite burden compared with placebo, drug-blank niosome, and MA (p < 0.05).Conclusion: Overall, these findings suggest that the NTB formulation exhibits enhanced pharmacological properties and effective antileishmanial activity, indicating the potential for further clinical research.Keywords: cutaneous leishmaniasis, niosomes, tioxolone-benzoxonium chloride, topical therapy, in silico analysis, in vivo efficacy