Development and neuroprotective testing of nanocarrier formulations for intranasal administration of curcumin derivatives CNB-001, J-147 and T-006

姜黄素是一种具有抗炎、抗氧化和神经保护特性的天然化合物。它可以减轻线粒体功能障碍，减少成人和新生儿缺血/再灌注和炎症模型的损伤。姜黄素临床应用的主要限制是其极低的水溶性（0.0004mg/ml）和生物利用度，缺乏确定的分子靶点及其无干扰特性。CNB-001， J-147和T-006是合成姜黄素类似物，因此允许生产和纯度的一致性。虽然这三种衍生物都有更好的分子靶点特征，并且已经在神经变性和中风中进行了试验，但它们具有低水溶性，并且溶解在DMSO中，这对人类是有毒的。此外，它们已被口服或静脉注射，这限制了通过全身消除给药到大脑，鼻内给药途径尚未实施。鼻内应用是非侵入性的，可以穿透血脑屏障，通过三叉神经和嗅觉途径更快地进入大脑，绕过外周清除。此外，鼻内给药比全身给药有效10倍，因此更具有临床意义。该项目旨在重新利用已经在临床试验中的药物，并开发用于微创鼻内应用的新型纳米颗粒制剂CNB-001，J-147和T-006。该项目将在体内评估最成功的配方的药代动力学，并在轮换项目中进行体外毒性测试。通过高效液相色谱法测定naïve新生小鼠和成年小鼠经鼻给药后CNB-001、J-147和T-006的血清和脑浓度。每种衍生物具有最佳药代动力学的配方将在新生儿缺氧缺血小鼠模型（Rocha-Ferreira et al., 2019）和成人面神经切开时进行组织学和行为学上的短期和长期体内神经保护评估，此前Hristova实验室已经观察到全身姜黄素给药的神经保护。

Curcumin is a natural compound with anti-inflammatory, anti-oxidant and neuroprotectivecharacteristics. It attenuates mitochondrial dysfunction and reduces damage in adult andneonatal ischemia/reperfusion and inflammation models. The major limitation to clinicalimplementation of curcumin is its extremely low water-solubility (0.0004mg/ml) andbioavailability, the lack of identified molecular target and its pan essay interferencecharacteristics. CNB-001, J-147 and T-006 are synthetic curcumin analogs, thus allowingconsistency in production and purity. Although all three derivatives have better characterizedmolecular targets and are already trialed in neurodegeneration and stroke, they possess lowwater solubility and dissolve in DMSO, which is toxic for humans. Additionally, they have beenapplied orally or intravenously, which limits delivery to the brain through systemicelimination, and intranasal route of administration has not been implemented. Intranasalapplication is non-invasive, allows penetration through the blood brain barrier, it is quicker forbrain delivery through the trigeminal and olfactory routes, and by-passes peripheralelimination. Moreover, intranasal delivery is 10-fold more effective than systemicadministration and is thus more clinically relevant. This project aims to repurpose drugs thatare already in clinical testing and to develop novel nanoparticle formulations of CNB-001,J-147 and T-006 for minimally invasive intranasal application.The project will assess in vivo the pharmacokinetics of the most successful formulationsgenerated and tested in vitro for toxicity in the rotation project. Serum and brainconcentration of CNB-001, J-147 and T-006 post-intranasal administration of the nanoparticleformulations applied in naïve neonatal and adult mice will be assessed through HPLC.The formulations with best pharmacokinetics for each derivative will be histologically andbehaviourally assessed for short- and long-term in vivo neuroprotection when appliedintranasally in mouse models of neonatal hypoxia-ischaemia (Rocha-Ferreira et al., 2019) andadult facial nerve axotomy, where previously the Hristova lab has observed neuroprotectionwith systemic curcumin administration.