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目的研究双修饰眼镜蛇神经毒素脂质体[(Pep2(Pep1)-αCT-LP)]鼻黏膜给药后药物在大鼠脑中的药动学行为。方法通过Michael加成反应合成二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep1及二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep2,通过核磁共振氢谱(~1H-NMR)和红外光谱(FTIR)验证产物结构。采用薄膜分散-后插入法制备双修饰眼镜蛇神经毒素脂质体,制得的脂质体考察形态、粒径、Zeta电位;采用超滤离心法测定脂质体包封率。应用大鼠脑微透析技术测定鼻黏膜给药后大鼠脑中枢中脑导水管周围灰质(periaqueductal gray,PAG)部位未修饰、单修饰以及双修饰眼睛蛇神经毒素脂质体药物浓度经时变化,用PKSolver软件处理药动学参数。结果核磁共振氢谱和红外光谱证实二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep1及二硬脂酰甘油磷脂乙醇胺-聚乙二醇-Pep2结构。制备的双修饰眼镜蛇神经毒素脂质体为球形或近球形,大小均一,粒径较小(115.8±1.86)nm,Zeta电位为(-13.77±0.75)m V,包封率为(32.75±1.12)%。在中脑导水管周围灰质部位,双修饰眼镜蛇神经毒素脂质体最大药物浓度(ρ_(max))显著高于其他各组脂质体(P<0.05),ρ_(max)、t_(max)、AUC_(0→∞)分别为(244.72±3.15)ng·mL~(-1)、(88.01±4.19)min、(89 199.02±1 922.99)ng·min·mL~(-1)。结论双修饰眼镜蛇神经毒素脂质体可以显著提高药物在中脑导水管周围灰质部位浓度,该结果为研究蛋白多肽类药物经鼻黏膜给药的脑靶向制剂的开发提供了参考。
OBJECTIVE: To study the pharmacokinetics of the drug in the rat brain after double-modification of nasal mucosa with modified zebra neurotoxin liposome [(Pep2 (Pep1) -αCT-LP)]. Methods Distearoylglycerophospholipid ethanolamine-polyethylene glycol-Pep1 and distearoyl-glycerophosphatidylethanolamine-polyethylene glycol-Pep2 were synthesized by Michael addition reaction. Their structures were characterized by 1H nuclear magnetic resonance (1H-NMR) and infrared Spectral (FTIR) validation of the product structure. Liposomes with double-modified cobra neurotoxin were prepared by membrane dispersion-post insertion method. The morphology, particle size and Zeta potential of the prepared liposomes were investigated. The entrapment efficiency of liposomes was determined by ultrafiltration centrifugation. Rat brain microdialysis technique was used to determine the concentration of unmodified, unmodified and double-modified zebra neurotoxin liposomes in the central cerebral pachycardia of rats after nasal mucosa administration for a time period , Using PKSolver software pharmacokinetic parameters. Results Nuclear magnetic resonance (1H) and infrared spectroscopy confirmed the distearoylglycerophospholipid ethanolamine-polyethylene glycol-Pep1 and distearoylglycerophosphatidylethanolamine-polyethylene glycol-Pep2 structures. The prepared double-modified serpentine neurotoxin liposomes were spherical or nearly spherical, with uniform size, smaller particle size (115.8 ± 1.86) nm, Zeta potential of (-13.77 ± 0.75) mV and encapsulation efficiency of (32.75 ± 1.12 )%. The maximal drug concentration (ρ max) of double-modified cobra neurotoxin liposomes was significantly higher than that of the other liposomes (P <0.05), ρ max, t max, , And the AUC_ (0 → ∞) were (244.72 ± 3.15) ng · mL -1, (88.01 ± 4.19) min and (89 199.02 ± 1 922.99) ng · min · mL -1, respectively. Conclusions Double modified cobra neurotoxin liposomes can significantly increase the concentration of drug in the midbrain periaqueductal gray, which provides a reference for the development of brain-targeting agents for the delivery of protein peptides via nasal mucosa.