Abstract：
Nitric oxide (NO) is a crucial gaseous signaling molecules in regulating cardiovascular, immune, and nervous systems. Controlled and targeted NO delivery is imperative for treating cancer, inflammation, and cardiovascular diseases. Despite various enzyme-prodrug therapy (EPT) systems facilitating controlled NO release, their clinical utility is hindered by nonspecific NO release and undesired metabolic consequence. In this study, a novel EPT system is presented utilizing a cellobioside-diazeniumdiolate (Cel2-NO) prodrug, activated by an endocellulase (Cel5A-h38) derived from the rumen uncultured bacterium of Hu sheep. This system demonstrates nearly complete orthogonality, wherein Cel2-NO prodrug maintains excellent stability under endogenous enzymes. Importantly, Cel5A-h38 efficiently processes the prodrug without recognizing endogenous glycosides. The targeted drug release capability of the system is vividly illustrated through an in vivo near-infrared imaging assay. The precise NO release by this EPT system exhibits significant therapeutic potential in a mouse hindlimb ischemia model, showcasing reductions in ischemic damage, ambulatory impairment, and modulation of inflammatory responses. Concurrently, the system enhances tissue repair and promotes function recovery efficacy. The novel EPT system holds broad applicability for the controlled and targeted delivery of essential drug molecules, providing a potent tool for treating cardiovascular diseases, tumors, and inflammation-related disorders.
摘要：
一氧化氮(NO)是调节心血管的重要气体信号分子,免疫，和神经系统。控制和靶向的NO递送对于治疗癌症至关重要，炎症,和心血管疾病。尽管各种酶-前药治疗（EPT）系统促进控制NO释放，非特异性NO释放和不良代谢后果阻碍了它们的临床应用。在这项研究中,提出了一种新的EPT系统，利用纤维二糖苷-二氮杂双醇盐（Cel2-NO）前药，由源自胡羊瘤胃未培养细菌的内纤维素酶（Cel5A-h38）激活。该系统显示出几乎完全的正交性，其中Cel2-NO前药在内源性酶下保持优异的稳定性。重要的是,Cel5A-h38有效地加工前药而不识别内源性糖苷。通过体内近红外成像测定生动地说明了该系统的靶向药物释放能力。这种EPT系统的精确NO释放在小鼠后肢缺血模型中显示出显著的治疗潜力。展示缺血性损伤的减少，动态障碍，和炎症反应的调节。同时,该系统增强组织修复并促进功能恢复功效。新的EPT系统对于基本药物分子的受控和靶向递送具有广泛的适用性。提供治疗心血管疾病的有力工具，肿瘤，和炎症相关的疾病。
