一种稳定的门冬胰岛素结晶制备方法的研究

摘要

目的：本文旨在初步研究出一种稳定的门冬胰岛素结晶制备的方法，优化生产工艺，降低生产成本，减少原料药生产过程中的损失，提高门冬胰岛素的稳定性，为制剂提供优质的原料药产品。方法在室温下经过有机酸对比，有机溶剂以及其他胰岛素结晶所必须添加的物质的选择，再经过一系列对比实验优化，并通过显微镜图片分析，要求晶型稳定均一性好，晶体形状规则；超高效液相色谱仪（HPLC）检测，各项指标符合药典标准，继而找到一种稳定且结晶收率高的方案。结果研究发现有机酸宜选用0.5M ～1.5M 的甘氨酸，同时采用体积分数12％～18％的乙腈、体积分数为0.2％～0.3％的苯酚、0.2～0.3M氯化钠溶液以及含2.0～5.0g· L －1重组门冬胰岛素，在初始pH为6.0～6.5的条件下加入醋酸锌组成的结晶液，经放大验证得到单斜针状晶体，形状规则均一，结晶收率可达99％以上。结论通过以上实验表明，门冬胰岛素原料药的结晶方法符合生产要求，结晶收率良好，易于工业化生产，可为制剂提供优质的原料药。%ABSTRACT:OBJECTIVE To study a kind of preparation method for the stable crystalline insulin aspart optimize production processes.Reduce production costs , reduce the loss of raw material medicine in production process , im-prove the stability of insulin aspart ,and provide high quality raw material medicine for the preparation.METHODS Compared by organic acid at room temperature.Selected the organic solvent and other insulin crystal material ,and then through a series of comparative experiments and optimization ,through the microscope image analysis for stable crystal form,good uniformity,crystal shape;ultra performance liquid chromatography ( HPLC) detection,the indica-tors are in line with the standard of Chinese Pharmacopoeia ,and then find a stable solution that the crystallization is of high yield.RESULTS The study found that organic acid should be selected as the best 0.5M~1.5M of glycine, at the same time ,using acetonitrile volume fraction of the volume fraction of 12%~18%,0.2~0.3%of phenol ,0.2~0.3 M Sodium Chloride Solution 2.0~5.0 g· L-1 containing recombinant insulin aspart ,crystallization solution at the initial pH to 6.0~6.5 under the conditions of adding zinc acetate.After amplification ,verification of monoclinic crystal regular shape ,uniform,the crystallization yield can reach more than 99%.CONCLUSION Through the a-bove experiments show that the method of crystallization ,insulin aspart raw materials meet the requirements of pro-duction,the yield of crystallization is good ,and it is easy for industrialized production ,can provide raw materials for preparation of high quality.