Abstract

Background

Intracarotid injection of mannitol has been applied for decades to support brain entry of therapeutics that otherwise do not effectively cross the blood–brain barrier (BBB). However, the elaborate and high-risk nature of this procedure has kept its use restricted to well-equipped medical centers. We are developing a more straightforward approach to safely open the BBB, based on the intra-arterial (IA) injection of NEO100, a highly purified version of the natural monoterpene perillyl alcohol.

Methods

In vitro barrier permeability with NEO100 was evaluated by transepithelial/transendothelial electrical resistance and antibody diffusion assays. Its mechanism of action was studied by western blot, microarray analysis, and electron microscopy. In mouse models, we performed ultrasound-guided intracardiac administration of NEO100, followed by intravenous application of Evan’s blue, methotrexate, checkpoint-inhibitory antibodies, or chimeric antigen receptor (CAR) T cells.

Results

NEO100 opened the BBB in a reversible and nontoxic fashion in vitro and in vivo. It enabled greatly increased brain entry of all tested therapeutics and was well tolerated by animals. Mechanistic studies revealed effects of NEO100 on different BBB transport pathways, along with translocation of tight junction proteins from the membrane to the cytoplasm in brain endothelial cells.

Conclusion

We envision that this procedure can be translated to patients in the form of transfemoral arterial catheterization and cannulation to the cerebral arteries, which represents a low-risk procedure commonly used in a variety of clinical settings. Combined with NEO100, it is expected to provide a safe, widely available approach to enhance brain entry of any therapeutic.

中文翻译：

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NEO100 可实现血脑屏障不可渗透疗法的大脑递送

摘要

背景

颈动脉内注射甘露醇已经应用了几十年，以支持治疗剂进入大脑，否则这些治疗剂不能有效地穿过血脑屏障 (BBB)。然而，该程序的复杂性和高风险性质使其使用仅限于设备齐全的医疗中心。我们正在开发一种更直接的方法来安全地打开 BBB，基于动脉内 (IA) 注射 NEO100，一种高度纯化的天然单萜紫苏醇。

方法

NEO100 的体外屏障通透性通过跨上皮/跨内皮电阻和抗体扩散试验进行评估。通过蛋白质印迹、微阵列分析和电子显微镜研究其作用机制。在小鼠模型中，我们在超声引导下对 NEO100 进行了心内给药，然后静脉注射了伊文蓝、甲氨蝶呤、检查点抑制性抗体或嵌合抗原受体 (CAR) T 细胞。

结果

NEO100 在体外和体内以可逆和无毒的方式打开 BBB。它大大增加了所有测试疗法的大脑进入率，并且动物耐受性良好。机理研究揭示了 NEO100 对不同 BBB 转运途径的影响，以及紧密连接蛋白从脑内皮细胞膜到细胞质的易位。

结论

我们设想该程序可以以经股动脉导管插入术和脑动脉插管的形式转化为患者，这是一种常用于各种临床环境的低风险程序。结合 NEO100，它有望提供一种安全、广泛可用的方法来增强任何治疗的大脑进入。