Lokhande Amit S, Maurya Vikas, Rani Komal, Parashar Palak, Gaind Rajni, Tandon Vibha, Devarajan Padma V
Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, N. P. Marg, Matunga, Mumbai 400019, Maharashtra, India.
Special Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi 110067, India.
Int J Pharm. 2024 Apr 25;655:123982. doi: 10.1016/j.ijpharm.2024.123982. Epub 2024 Mar 8.
Recently, World Health Organization declared antimicrobial resistance as the third greatest threat to human health. Absence of known cross-resistance, new class, new target, and a new mode of action are few major strategies being undertaken by researches to combat multidrug resistant pathogen. PPEF.3HCl, a bisbenzimidazole was developed as highly potent antibacterial agent against ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogens, targeting topoisomerase IA. The present work encompasses a radical on-site generation of In-situ nanosuspension of PPEF.3HCl with enhanced efficacy against methicillin resistant S. aureus in septicemia model. We have generated instantaneously a PPEF.3HCl nanosuspension (IsPPEF.3HCl-NS) by mixing optimized monophasic PPEF.3HCl preconcentrate in propylene glycol into an aqueous medium comprising tween 80 as stabilizer. The IsPPEF.3HCl-NS showed precipitation efficiency of > 90 %, average particle size < 500 nm, retained upto 5 h, a negative zeta potential and bi/trimodal particle size distribution. Differential scanning calorimetry, X-ray diffraction confirmed partial amorphization and transmission electron microscopy revealed spherical particles. IsPPEF.3HCl-NS was non-hemolytic and exhibited good stability in serum. More significantly, it exhibited a ∼ 1.6-fold increase in macrophage uptake compared to free PPEF.3HCl in the RAW 264.7 macrophage cell line. Confocal microscopy revealed accumulation of IsPPEF.3HCl-NS within the lysosomal compartment and cell cytosol, proposing high efficacy. In terms of antimicrobial efficacy, IsPPEF.3HCl-NS outperforms free PPEF.3HCl against clinical methicillin sensitive and resistant S. aureus strains. In a pivotal experiment, IsPPEF.3HCl-NS exhibited over 83 % survival at 8 mg/kg.bw and an impressive reduction of ∼ 4-5 log-fold in bacterial load, primarily in the kidney, liver and spleen of septicemia mice. IsPPEF.3HCl-NS prepared by the In-situ approach, coupled with enhanced intramacrophage delivery and superior efficacy, positions IsPPEF.3HCl-NS as a pioneering and highly promising formulation in the battle against antimicrobial resistance.
最近，世界卫生组织宣布抗菌药物耐药性是对人类健康的第三大威胁。缺乏已知的交叉耐药性、新类别、新靶点和新作用模式是研究人员为对抗多重耐药病原体而采取的一些主要策略。PPEF.3HCl是一种双苯并咪唑，被开发为针对ESKAPE（粪肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌属）病原体的高效抗菌剂，其作用靶点为拓扑异构酶IA。目前的工作包括在败血症模型中通过自由基原位生成PPEF.3HCl的纳米悬浮液，其对耐甲氧西林金黄色葡萄球菌的疗效增强。我们通过将优化的单相PPEF.3HCl预浓缩物与丙二醇混合到含有吐温80作为稳定剂的水性介质中，瞬间生成了PPEF.3HCl纳米悬浮液（IsPPEF.3HCl-NS）。IsPPEF.3HCl-NS的沉淀效率>90%，平均粒径<500nm，可保持5小时，具有负的zeta电位和双/三峰粒径分布。差示扫描量热法、X射线衍射证实了部分非晶化，透射电子显微镜显示为球形颗粒。IsPPEF.3HCl-NS无溶血作用，在血清中表现出良好的稳定性。更显著的是，与RAW 264.7巨噬细胞系中的游离PPEF.3HCl相比，它的巨噬细胞摄取增加了约1.6倍。共聚焦显微镜显示IsPPEF.3HCl-NS在溶酶体区室和细胞质中积累，表明其具有高效性。在抗菌效果方面，IsPPEF.3HCl-NS在对抗临床甲氧西林敏感和耐药金黄色葡萄球菌菌株方面优于游离PPEF.3HCl。在一项关键实验中，IsPPEF.3HCl-NS在8mg/kg体重时的存活率超过83%，细菌载量显著降低约4-5个对数级，主要在败血症小鼠的肾脏、肝脏和脾脏中。通过原位方法制备的IsPPEF.3HCl-NS，结合增强的巨噬细胞内递送和卓越的疗效，使其在对抗抗菌药物耐药性的斗争中成为一种开创性且极具前景的制剂。
