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

姜黄素是一种天然化合物，具有抗炎、抗氧化和神经保护作用。在成人和新生儿缺血/再灌注和炎症模型中，它可减轻线粒体功能障碍并减少损伤。姜黄素的水溶性极低（0.0004mg/ml），生物利用度低，缺乏可识别的分子靶点，且具有广谱的干扰特性，是限制其临床应用的主要因素。CNB-001、J-147和T-006是合成的姜黄素类似物，因此在生产和纯度方面具有一致性。虽然这三种衍生物都有更好的分子靶点，并且已经在神经退行性疾病和中风中进行了试验，但它们具有低水溶性，并溶于对人类有毒的DMSO中。此外，它们已被口服或静脉内应用，这限制了通过全身消除向大脑的递送，并且尚未实施鼻内给药途径。鼻内给药是非侵入性的，可以穿透血脑屏障，通过三叉神经和嗅觉途径更快地向大脑递送，并绕过外周消除。此外，鼻内给药比全身给药有效10倍，因此更具有临床意义。本项目旨在重新利用已在临床试验中的药物，开发CNB-001、J-147和T-006的新型纳米粒制剂，用于微创鼻内应用。本项目将在体内评估最成功的制剂的药代动力学，并在轮换项目中进行体外毒性测试。将通过HPLC评估鼻内施用纳米颗粒制剂后CNB-001、J-147和T-006在未处理的新生小鼠和成年小鼠中的血清和脑浓度。将在组织学和行为学上评估每种衍生物具有最佳药代动力学的制剂在鼻内施用于新生儿缺氧-缺血小鼠模型中时的短期和长期体内神经保护作用（Rocha-Ferreira et al.，2019）和成人面神经轴突切断术，之前Hristova实验室已经观察到全身姜黄素给药的神经保护作用。

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.