新型纳豆激酶-γ-聚谷氨酸复合物注射液的制备和毒性评价  

Preparation and Toxicity Evaluation of a Novel Nattokinase-γ-polyglutamic Acid Complex Injection

作　　者：李静 王梦静[2] 陈杰鹏 段丽丽 骆翔[2] 邓意辉[2] 宋艳志[2] LI Jing;WANG Mengjing;CHEN Jiepeng;DUAN Lili;LUO Xiang;DENG Yihui;SONG Yanzhi(The Third Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China;School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China;Sungen Biotech Co.,Ltd.,Shantou 515000,China)

机构地区：[1]锦州医科大学附属第三医院,辽宁锦州121000 [2]沈阳药科大学药学院,沈阳110016 [3]广东双骏生物科技有限公司,广东汕头515000

出　　处：《中国现代应用药学》2020年第2期180-186,共7页Chinese Journal of Modern Applied Pharmacy

摘　　要：目的制备一种低毒性的纳豆激酶(nattokinase,NK)注射液。方法在NK注射液中加入γ-聚谷氨酸(γ-polyglutamic acid,γ-PGA),制备NK-γ-PGA复合物注射液。采用急性毒性试验,以小鼠给药后的生存时间、生存状态和注射后48h存活小鼠尾部的状态作为毒性评价的指标,分别考察pH和γ-PGA浓度对NK与γ-PGA复合情况的影响。利用体外溶栓试验对NK-γ-PGA注射液毒性降低的原因进行分析。结果当以120 kU·kg^-1的剂量单次尾静脉注射NK-γ-PGA复合物注射液(NK 10 kU·mL^-1,γ-PGA 0.924 mg·mL^-1,pH 6.0)时,48 h后小鼠的死亡率为0,且尾部无红肿及坏死,说明在pH 6.0和γ-PGA的浓度为0.924mg·mL^-1时,NK与γ-PGA复合得最好。体外溶栓试验结果表明,γ-PGA的加入未降低NK活性,毒性的降低主要是因为NK与γ-PGA形成了复合物,起到了缓释作用。结论本研究中制备的NK-γ-PGA复合物注射液(NK 10 kU·mL^-1,γ-PGA 0.924 mg·mL^-1,pH 6.0)在不改变NK活性的前提下,降低了NK注射液毒性。OBJECTIVE To prepare a low toxicity nattokinase(NK) injection.METHODS By adding the γ-polyglutamic acid(γ-PGA) to the NK solution,the NK-γ-PGA complex injection was prepared.Taking survival time,survival state and the tail state of the surviving mouse at 48 h after a single tail intravenous injection as toxicity evaluation indexes,the acute toxicity test was used to study the effects of pH and γ-PGA concentration on the complexation of NK and γ-PGA separately.The in vitro thrombolytic test was used to analyze the reason of the toxicity reduction of NK-γ-PGA complex injection.RESULTS When the NK-γ-PGA complex injection(NK 10 kU·mL^-1,γ-PGA 0.924 mg·mL^-1,pH 6.0) was administrated by a single tail intravenous injection at a dose of 120 kU·kg^-1,the death rate of mice was 0,and the mouse tail had no swelling and necrosis,which indicated that when the pH was 6.0 and the concentration of γ-PGA was 0.924 mg·mL^-1,the complexation of NK and γ-PGA was the best.The results of in vitro thrombolytic test showed that the addition of γ-PGA did not reduce the activity of the NK,but had a sustained-release effect to reduce drug toxicity.CONCLUSION The NK-γ-PGA complex injection(NK 10 kU·mL^-1,γ-PGA 0.924 mg·mL^-1,pH 6.0) prepared in this study achieved the goal of reducing NK injection toxicity without changing NK activity.

关 键 词：纳豆激酶 注射液 聚谷氨酸 复合物 急性毒性试验 体外溶栓试验 

分 类 号：R944.1[医药卫生—药剂学]