重组水蛭素-2鼻腔黏附纳米粒及其冻干粉药代动力学和抗凝活性研究

目的: 对重组水蛭素-2(rHV2)鼻腔壳聚糖(CS)黏附纳米粒及其冻干粉进行药代动力学和抗凝活性评价。 方法: 采用呈色肽比色法检测rHV2血药浓度;以正常麻醉大鼠为模型,测定经鼻腔给予rHV2生理盐水溶液、rHV2纳米粒溶液及其纳米粒冻干粉复溶液的药-时曲线,计算药代动力学参数及与rHV2生理盐水溶液皮下注射给药的相对生物利用度;同时以部分凝血活酶时间(APTT)为指标比较它们的抗凝活性。 结果: rHV2纳米粒比其生理盐水溶液鼻腔给药的相对生物利用度提高了8.99倍,rHV2纳米粒冻干粉复溶液也较其生理盐水溶液鼻腔给药的相对生物利用度提高了8.48倍。rHV2纳米粒及其冻干粉鼻腔给药后,均表现出显著的抗凝血活性,而rHV2生理盐水鼻腔给药则基本无效。 结论: 制备的rHV2鼻腔壳聚糖黏附纳米粒制剂具有较高的生物利用度,制成冻干粉后,仍保持较高的生物利用度。由于rHV2冻干粉制剂稳定性更高,有望成为rHV2经鼻腔给药的有效制剂。

Objective: To evaluate of pharmacokinetic and anticoagulant activity on recombinant hirudin-2(rHV2)bio-adhesive nanoparticles and its frozen-dried powder for nasal delivery. Methods: Healthy SD male rats were divided into rHV2 sailine solution subcutaneous administration(s.c.)group, rHV2 sailine solution intranasal(i.n.)administration group, rHV2 nanoparticles intranasal administration(i.n.)group and rHV2 nanoparticles frozen-dried powder intranasal administration(i.n.)group, 5-6 rats for each group. rHV2 concentration in plasma was determined by chromogenic substrate method. Compared to rHV2 sailine solution s.c. administration group, the relative bioavailabilities(F_r)and pharmacokinetic parameters of each rHV2 i.n. administration groups were calculated by the software DAS. Taking activated partial thromboplastin time(APTT)as the index and rHV2 sailine solution s.c. administration groups, the anticoagulation effects of above groups were investigated. Results: rHV2 chitosan nanoparticles and its frozen-dried powder could improve F_r of rHV2 i.n. administration greatly. F_r of two preparations respectively increased 8.99 times and 8.48 times more than rHV2 sailine solution i.n. administration. Significant effects of anticoagulation were observed in normal rats after i.n. administration of rHV2 nanoparticles and its frozen-dried powder. Conclusion: rHV2 chitosan nanoparticles and its frozen-dried powder showed higher bioavailability following i.n. administration, so they should be a better nasal preparation of rHV2.

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