Abstract：
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.
摘要：
可注射抗凝剂广泛用于医疗程序中以防止不需要的血液凝固。然而,许多人缺乏安全，有效的逆转剂。这里,我们提供了关于先前描述的基于RNA折纸的新数据，直接凝血酶抑制剂(HEX01)。我们描述了一个新的，快速行动，具体,单分子逆转剂（解毒剂），并首次提供体内数据，包括功效，可逆性,初步安全,和初步的生物分布研究。HEX01包含附加在RNA折纸上的多个结合凝血酶的适体。它在体外和体内表现出优异的抗凝血活性。新的单分子,DNA解毒剂(HEX02)在体外30秒内逆转人血浆中HEX01的抗凝活性,并且在鼠肝裂伤模型中有效地起作用。使用离体成像的HEX01在整个小鼠中的生物分布研究显示,在24小时内主要在肝脏中积累,并且在肾脏中浓度降低10倍。此外,我们显示HEX01/HEX02系统对上皮细胞系无细胞毒性且在体外无溶血性。此外，我们在小鼠模型中没有发现血清细胞因子对HEX01/HEX02的反应。HEX01和HEX02代表安全有效的凝血控制系统，具有快速作用，特定的逆转剂显示出潜在药物开发的希望。
