Manrique-Suárez Viana, Mangui Catota Bryan A, Camacho Casanova Frank, Jara Mendoza Nery A, Contreras Vera Maria A, Maura Pérez Rafael, Reyes López Fátima, Toledo Alonso Roberto, Castro Henriquez Pablo Ignacio, Sánchez Ramos Oliberto
Pharmacology Department, School of Biological Sciences, Recombinant Biopharmaceuticals Laboratory, University of Concepcion, Concepcion, Chile.
Pharmacology Department, School of Biological Sciences, Cellular Pharmacology Laboratory, University of Concepcion, Concepcion, Chile.
J Drug Target. 2025 Apr;33(4):546-555. doi: 10.1080/1061186X.2024.2434908. Epub 2024 Dec 2.
Effective drug delivery to the central nervous system (CNS) remains a challenge due to the blood-brain barrier (BBB). Macromolecules such as proteins and peptides are unable to cross BBB and have poor therapeutic efficacy due to little or no drug distribution. A promising alternative is the conjugation of a drug to a shuttle molecule that can reach the CNS via receptor-mediated transcytosis (RMT). Several receptors have been described for RMT, such as low-density lipoprotein receptor-related protein 1 (LRP1). We used phage display technology combined with an BBB model to identify LRP1 ligands. A single domain antibody (dAb) library was used to enrich for species that selectively bind to immobilised LRP1 ligand. We obtained a novel nanobody, dAb D11, that selectively binds to LRP1 receptor and mediates internalisation of phage particles in brain endothelial cells, with a dissociation constant Kd of 183.1 ± 85.8 nM. The high permeability of D11 was demonstrated by an biodistribution assay in mice. We discovered D11, the first LRP1 binding dAb with BBB permeability. Our findings will contribute to the development of RMT-based drugs for the treatment of CNS diseases.
由于血脑屏障（BBB）的存在，将药物有效递送至中枢神经系统（CNS）仍然是一项挑战。蛋白质和肽等大分子无法穿过血脑屏障，且由于药物分布极少或没有分布，治疗效果不佳。一种有前景的替代方法是将药物与穿梭分子偶联，该穿梭分子可通过受体介导的转胞吞作用（RMT）到达中枢神经系统。已经描述了几种用于RMT的受体，如低密度脂蛋白受体相关蛋白1（LRP1）。我们使用噬菌体展示技术结合血脑屏障模型来鉴定LRP1配体。一个单域抗体（dAb）文库用于富集选择性结合固定化LRP1配体的物种。我们获得了一种新型纳米抗体dAb D11，它选择性地结合LRP1受体并介导噬菌体颗粒在内皮细胞中的内化，解离常数Kd为183.1±85.8 nM。通过在小鼠体内的生物分布试验证明了D11的高渗透性。我们发现了D11，第一个具有血脑屏障渗透性的LRP1结合dAb。我们的发现将有助于开发基于RMT的治疗中枢神经系统疾病的药物。
