Department of Pharmaceutics, Faculty of Pharmacy, The Maharaja Sayajirao University of Baroda, Kalabhavan Campus, Vadodara 390001, Gujarat, India.
Department of Pharmaceutics, Faculty of Pharmacy, The Maharaja Sayajirao University of Baroda, Kalabhavan Campus, Vadodara 390001, Gujarat, India; Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, 12901 Bruce B. Downs Blvd., MDC30, Tampa, FL 33612, United States.
Int J Pharm. 2019 May 30;563:324-336. doi: 10.1016/j.ijpharm.2019.04.006. Epub 2019 Apr 5.
Rational design of novel ionizable lipids for development of lipid-nucleic acid nanoparticles (LNP) is required for safe and effective systemic gene delivery for osteoporosis. LNPs require suitable characteristics for intravenous administration and effective accumulation in bone marrow for enhanced transfection. Hence, lipids with C18 tail and ionizable headgroup (Boc-His-ODA/BHODA and His-ODA/HODA) were synthesized and characterized physicochemically. LNPs were prepared with bone morphogenetic protein-9 gene (BHODA-LNP, HODA-LNP, and bone-homing peptide targeted HODA-LNP - HODA-LNPT). Thorough physicochemical (electrolyte stability, DNase I and serum stability) and biological (hemolysis, ROS induction, cytotoxicity and transfection) characterization was carried out followed by acute toxicity studies and therapeutic performance studies in ovariectomized rat model. Lipids with pH dependent ionization were successfully synthesized. LNPs thereof were ∼100 nm size with stability against electrolytes, DNase I and serum and exhibited low hemolytic potential demonstrating suitability for intravenous administration. LNPs exhibited minimal cytotoxicity, non-significant ROS induction and high transfection. In vivo studies demonstrated safety and improved bone regeneration in OVX rats with HODA-LNPT showing significantly better performance. Synthesized ionizable lipids offer safe and effective alternative for preparation of LNPs for gene delivery. Targeted BMP-9 LNP show potential for systemic osteoporosis treatment.
需要设计新型可离子化脂质以用于开发用于骨质疏松症的安全有效的全身基因传递的脂质 - 核酸纳米颗粒（LNP）。LNPs 需要适合静脉内给药的特性，并在骨髓中有效积累以增强转染。因此，合成了具有 C18 尾部和可离子化头基（Boc-His-ODA/BHODA 和 His-ODA/HODA）的脂质，并对其进行了理化特性的表征。用骨形态发生蛋白-9 基因（BHODA-LNP、HODA-LNP 和靶向骨归肽的 HODA-LNP-HODA-LNPT）制备了 LNP。进行了彻底的理化（电解质稳定性、DNase I 和血清稳定性）和生物学（溶血、ROS 诱导、细胞毒性和转染）表征，随后进行了急性毒性研究和去卵巢大鼠模型的治疗性能研究。成功合成了具有 pH 依赖性离解的脂质。其 LNP 的大小约为 100nm，具有对电解质、DNase I 和血清的稳定性，并且溶血潜力低，表明适合静脉内给药。LNP 表现出低细胞毒性、无显著 ROS 诱导和高转染。体内研究表明，HODA-LNPT 在 OVX 大鼠中具有安全性和改善的骨再生，表现出更好的性能。合成的可离子化脂质为制备用于基因传递的 LNP 提供了安全有效的替代方法。靶向 BMP-9 LNP 具有用于全身骨质疏松症治疗的潜力。
