Cyclodextrin-AmB-IL-10 antagonist peptide nanoparticles treat leishmaniasis more effectively than conventional AmB

This study aimed to develop a novel nanotherapeutic approach by combining an interleukin-10 (IL-10) peptide antagonist with amphotericin B (AmB) to enhance antileishmanial efficacy while reducing cytotoxicity. A peptide antagonist targeting IL-10, identified via <i>in-silico</i> analysis and showing minimal cytotoxicity (95% promastigote viability at 20 µg/mL), was synthesized and conjugated with AmB using chemical cross-linkers. The conjugate was encapsulated in gamma-cyclodextrin to produce uniform nanoparticles (~40 nm). Characterization was performed using Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). In vitro and in vivo studies were conducted to assess parasite burden, immune response, and safety parameters. The nanoformulation significantly reduced parasite burden and improved amastigote clearance, as confirmed by real-time PCR. Cytokine profiling revealed elevated IL-12 and other protective cytokines, indicating enhanced immune modulation. Hematological, biochemical, and splenomegaly analyses demonstrated improved safety and therapeutic efficacy compared to AmB alone. The IL-10 antagonist – AmB nanoformulation represents a promising immunomodulatory therapeutic strategy for leishmaniasis. Its enhanced efficacy and safety highlight its potential for application, particularly in immunocompromised patients.

本研究旨在开发一种新型纳米治疗策略，将白细胞介素-10（interleukin-10, IL-10）肽拮抗剂与两性霉素B（amphotericin B, AmB）联合应用，以提升抗利什曼原虫疗效同时降低细胞毒性。研究通过虚拟（in-silico）分析筛选得到靶向IL-10的肽拮抗剂，该拮抗剂在20 μg/mL浓度下对利什曼原虫前鞭毛体的存活率影响极小（存活率达95%）；随后完成该肽拮抗剂的合成，并通过化学交联剂将其与AmB偶联。将所得偶联物包封于γ-环糊精中，制备得到粒径约40 nm的均一纳米颗粒。采用傅里叶变换红外光谱（Fourier Transform Infrared Spectroscopy, FTIR）、扫描电子显微镜（Scanning Electron Microscopy, SEM）及透射电子显微镜（Transmission Electron Microscopy, TEM）对纳米制剂进行表征。通过体外与体内实验评估寄生虫载量、免疫应答及安全性相关参数。实时荧光定量PCR结果证实，该纳米制剂可显著降低寄生虫载量并促进无鞭毛体清除。细胞因子谱分析显示IL-12及其他保护性细胞因子水平显著升高，表明免疫调控作用得以增强。血液学检测、生化指标分析及脾脏肿大评估结果表明，相较于单独使用AmB，该纳米制剂的安全性与治疗功效均得到显著改善。IL-10拮抗剂-AmB纳米制剂为利什曼病提供了一种极具前景的免疫调节治疗策略，其增强的疗效与安全性凸显了其临床应用潜力，尤其适用于免疫功能低下患者。