LEA蛋白特征片段及海藻糖对热敏性蛋白药物胰岛素冻干的协同保护

发布时间：2018-12-08 11:35

【摘要】：热敏性蛋白药物胰岛素的热稳定性较差,极易受制备方式和保存条件等影响而失活。故本研究分别利用高效液相色谱(HPLC)测定冻干样品中胰岛素的效价和差示扫描量热仪(DSC)测定冻干样品的玻璃化转变温度等重要参数,来研究胚胎发育晚期富集蛋白特征片段(LEA-motif,LEAM)和海藻糖冻干保护剂对胰岛素冻干保护效果的影响。冻干实验表明,两种保护剂均对胰岛素的活性具有保护效果,且LEAM的活性保护效果优于海藻糖。在效价和热稳定性方面,LEAM保护的胰岛素冻干品均高于海藻糖保护的胰岛素冻干样品。更为重要的是,两种保护剂协同保护时,少量海藻糖对于LEAM的保护特性具有增效作用。在冻干过程中,可有效提升药物玻璃态的稳定性,防止热致变性失活。由此可见,这种LEAM和海藻糖保护剂复合保护方法有望也可以应用在热敏性蛋白药物冻干粉针制备过程以保护药物活性。
[Abstract]:The thermal stability of heat sensitive protein drug insulin is poor, and it is easily inactivated due to the influence of preparation methods and preservation conditions. In this study, the titer of insulin in freeze-dried samples was determined by high performance liquid chromatography (HPLC) and the glass transition temperature of freeze-dried samples was determined by differential scanning calorimeter (DSC). To study the effects of LEA-motif,LEAM and trehalose lyophilization protectant on the protective effect of insulin freeze-drying. The freeze-drying experiments showed that both of the two protective agents had protective effects on the activity of insulin, and the protective effect of LEAM was better than that of trehalose. In terms of titer and thermal stability, LEAM protected insulin freeze-dried products were higher than trehalose protected insulin freeze-dried samples. More importantly, a small amount of trehalose has a synergistic effect on the protective properties of LEAM. In the freeze-drying process, the stability of the glass state of the drug can be improved effectively, and the thermal denaturation and inactivation can be prevented. Therefore, the composite protection method of LEAM and trehalose protectant may also be used in the preparation of freeze dried powder for thermosensitive protein drugs to protect the drug activity.
【作者单位】：上海理工大学生物系统热科学研究所;上海东富龙科技股份有限公司;
【基金】：国家自然科学基金(51076108)资助项目上海市“创新行动计划”国际科技合作项目(12430702000) 上海市重点学科项目(T0503和P0502) 上海市自然科学基金(12ZR1420400) 上海市教委科研创新项目(14YZ092) 上海市联盟计划项目资助~~
【分类号】：R943

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