Ahmad Niyaz, Ahmad Rizwan, Ahmad Farhan Jalees, Ahmad Wasim, Alam Md Aftab, Amir Mohd, Ali Abuzer
Department of Pharmaceutics, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.
Department of Pharmaceutical Chemistry, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.
Saudi J Biol Sci. 2020 Jan;27(1):500-517. doi: 10.1016/j.sjbs.2019.11.008. Epub 2019 Nov 22.
Here, the aim is to improve the bioavailability of Naringenin (NRG) in brain and to establish the highest remedial benefit from a novel anti-ischemic medicine i.e. NRG.
A novel Naringenin-loaded-nanoemulsion (NE)-()-gel (i.e. thermoresponsive), was formulated with the help of Poloxamer-407 (20.0% w/v). Chitosan (CS, 0.50% w/v) was used to introduce the mucoadhesive property of NE-()-gel and finally called as NRG-NE-gel + 0.50%CS. A novel UHPLC-ESI-Q-TOF-MS/MS-method was optimized and used for NRG-NE-gel + 0.50%CS to quantify the Pharmacokinetic-(PK)-parameters in plasma as well as brain and to evaluate the cerebral ischemic parameters after MCAO i.e. locomotor activity, grip strength, antioxidant activity, and quantity the infarction volume in neurons with the safety/toxicity of NRG-NE-gel + 0.50%CS after i.n. administration in the rats.
The mucoadhesive potency and gelling temperature of NRG-NE-gel + 0.50%CS were observed 6245.38 dynes/cm and 28.3 ± 1.0 °C, respectively. Poloxamer-407 based free micelles size was observed 98.31 ± 1.17 nm with PDI (0.386 ± 0.021). The pH and viscosity of NRG-NE-gel + 0.50%CS were found to be 6.0 ± 0.20 and 2447 ± 24cp (at 35.0 ± 1.0 °C temperature), respectively. An elution time and / NRG were observed 1.78 min and 270.97/150.96 with 1.22 min and / of 301.01/150.98 for Quercetin (IS) respectively. Inter and intra %precision and %accuracy was validated 1.01-3.37% and 95.10-99.30% with a linear dynamic range (1.00 to 2000.00 ng/ml). AUC of plasma & brain were observed 995.60 ± 24.59 and 5600.99 ± 144.92 (ng min/ml g) in the rats after the intranasal (i.n.) administration of NRG-NE-gel + 0.50%CS. No toxicological response were not found in terms of mortalities, any-change morphologically i.e. in the microstructure of brain as well as nasal mucosa tissues, and also not found any visual signs in terms of inflammatory or necrosis.
Intranasally administered NRG-NE-gel + 0.50%CS enhanced the bioavailability of Naringenin in the brain. In the cerebral ischemic rats, significantly improved the neurobehavioral activity (locomotor & grip strength) followed by antioxidant activity as well as infarction volume. Finally, the toxicity studies carried out and established the safe nature of optimized-NRG-NE-gel + 0.50%CS.
本研究旨在提高柚皮素（NRG）在脑中的生物利用度，并确定一种新型抗缺血药物即NRG的最大治疗益处。
借助泊洛沙姆-407（20.0% w/v）制备了一种新型载柚皮素纳米乳（NE）-（）-凝胶（即热响应性凝胶）。壳聚糖（CS，0.50% w/v）用于赋予NE-（）-凝胶粘膜粘附特性，最终称为NRG-NE-凝胶+0.50%CS。优化了一种新型超高效液相色谱-电喷雾-四极杆-飞行时间-串联质谱法（UHPLC-ESI-Q-TOF-MS/MS），用于测定NRG-NE-凝胶+0.50%CS在血浆和脑中的药代动力学（PK）参数，并评估大脑中动脉闭塞（MCAO）后的脑缺血参数，即运动活性、握力、抗氧化活性，以及在大鼠经鼻内给药后，对NRG-NE-凝胶+0.50%CS的安全性/毒性进行评估，定量神经元中的梗死体积。
观察到NRG-NE-凝胶+0.50%CS的粘膜粘附力和胶凝温度分别为6245.38达因/厘米和28.3±1.0°C。基于泊洛沙姆-407的游离胶束大小为98.31±1.17纳米，多分散指数（PDI）为0.386±0.021。发现NRG-NE-凝胶+0.50%CS的pH值和粘度分别为6.0±0.20和2447±24厘泊（在35.0±1.0°C温度下）。观察到洗脱时间和/NRG分别为1.78分钟和2分别为1.22分钟和/槲皮素（内标）的301.01/150.98。批内和批间的%精密度和%准确度在1.01-3.37%和95.10-99.30%范围内得到验证，线性动态范围为（1.00至2000.00纳克/毫升）。在大鼠经鼻内（i.n.）给予NRG-NE-凝胶+0.50%CS后，观察到血浆和脑的AUC分别为995.60±24.59和5600.99±144.92（纳克·分钟/毫升·克）。在死亡率、形态学任何变化（即脑和鼻粘膜组织的微观结构）方面均未发现毒理学反应，在炎症或坏死方面也未发现任何可见迹象。
经鼻内给药的NRG-NE-凝胶+0.50%CS提高了柚皮素在脑中的生物利用度。在脑缺血大鼠中，显著改善了神经行为活性（运动和握力），随后是抗氧化活性以及梗死体积。最后，进行了毒性研究并确定了优化后的NRG-NE-凝胶+0.50%CS的安全性。
