Department of Materials Science and Engineering, College of Engineering, North Carolina State University, Raleigh, NC 27695, USA.
Joint Department of Biomedical Engineering, College of Engineering, North Carolina State University and University of North Carolina-Chapel Hill, Raleigh, NC 27695, USA.
Mol Ther. 2024 Jul 3;32(7):2286-2298. doi: 10.1016/j.ymthe.2024.05.002. Epub 2024 May 7.
Injectable anticoagulants are widely used in medical procedures to prevent unwanted blood clotting. However, many lack safe, effective reversal agents. Here, we present new data on a previously described RNA origami-based, direct thrombin inhibitor (HEX01). We describe a new, fast-acting, specific, single-molecule reversal agent (antidote) and present in vivo data for the first time, including efficacy, reversibility, preliminary safety, and initial biodistribution studies. HEX01 contains multiple thrombin-binding aptamers appended on an RNA origami. It exhibits excellent anticoagulation activity in vitro and in vivo. The new single-molecule, DNA antidote (HEX02) reverses anticoagulation activity of HEX01 in human plasma within 30 s in vitro and functions effectively in a murine liver laceration model. Biodistribution studies of HEX01 in whole mice using ex vivo imaging show accumulation mainly in the liver over 24 h and with 10-fold lower concentrations in the kidneys. Additionally, we show that the HEX01/HEX02 system is non-cytotoxic to epithelial cell lines and non-hemolytic in vitro. Furthermore, we found no serum cytokine response to HEX01/HEX02 in a murine model. HEX01 and HEX02 represent a safe and effective coagulation control system with a fast-acting, specific reversal agent showing promise for potential drug development.
注射用抗凝剂广泛用于医疗程序中，以防止不必要的血栓形成。然而，许多抗凝剂缺乏安全、有效的逆转剂。在这里，我们提出了一种新的基于 RNA 折纸的、直接凝血酶抑制剂（HEX01）的新数据。我们描述了一种新的、快速作用、特异性、单分子逆转剂（解毒剂），并首次提供了体内数据，包括疗效、逆转性、初步安全性和初步体内分布研究。HEX01 包含多个附在 RNA 折纸上的凝血酶结合适体。它在体外和体内均表现出优异的抗凝活性。新的单分子 DNA 解毒剂（HEX02）可在 30 秒内在人血浆中逆转 HEX01 的抗凝活性，并在小鼠肝裂伤模型中有效发挥作用。使用离体成像在整个小鼠中进行的 HEX01 的体内分布研究表明，其在 24 小时内主要在肝脏中积累，在肾脏中的浓度低 10 倍。此外，我们表明 HEX01/HEX02 系统对上皮细胞系无细胞毒性，且在体外无溶血作用。此外，我们在小鼠模型中未发现血清细胞因子对 HEX01/HEX02 的反应。HEX01 和 HEX02 代表了一种安全有效的凝血控制系统，具有快速作用、特异性逆转剂，为潜在药物开发提供了希望。
