Poulsen Christian, Jacobsen Dorte, Palm Lisbeth
Pharmaceutics, Diabetes Department, Novo Nordisk A/S, Novo Alle, room 6B1.58.1, 2880 Bagsvaerd, Denmark.
Pharm Res. 2008 Nov;25(11):2534-44. doi: 10.1007/s11095-008-9670-8. Epub 2008 Jul 8.
To examine the effect of different amine compounds on the chemical degradation of insulin aspart at pharmaceutical formulation conditions.
Insulin aspart preparations containing amine compounds or phosphate (reference) were prepared and the chemical degradation was assessed following storage at 37 degrees C using chromatographic techniques. Ethylenediamine was examined at multiple concentrations and the resulting insulin-ethylenediamine derivates were structurally characterized using matrix assisted laser desorption ionization time-of-flight mass spectroscopy. The effects on ethylenediamine when omitting glycerol or phenolic compounds from the formulations were investigated.
Ethylenediamine was superior in terms of reducing formation of high molecular weight protein and insulin aspart related impurities compared to the other amine compounds and phosphate. Monotransamidation of insulin aspart in the presence of ethylenediamine was observed at all of the six possible Asn/Gln residues with Asn(A21) having the highest propensity to react with ethylenediamine. Data from formulations studies suggests a dual mechanism of ethylenediamine and a mandatory presence of phenolic compounds to obtain the effect.
The formation of high molecular weight protein and insulin aspart related impurities was reduced by ethylenediamine in a concentration dependant manner.
考察不同胺类化合物在药物制剂条件下对门冬胰岛素化学降解的影响。
制备含胺类化合物或磷酸盐（对照）的门冬胰岛素制剂，并使用色谱技术评估在37℃储存后的化学降解情况。对多种浓度的乙二胺进行考察，并使用基质辅助激光解吸电离飞行时间质谱对所得的胰岛素-乙二胺衍生物进行结构表征。研究了制剂中省略甘油或酚类化合物时对乙二胺的影响。
与其他胺类化合物和磷酸盐相比，乙二胺在减少高分子量蛋白质和门冬胰岛素相关杂质的形成方面表现更优。在乙二胺存在下，观察到门冬胰岛素在所有六个可能的天冬酰胺/谷氨酰胺残基处发生单酰胺化，其中天冬酰胺（A21）与乙二胺反应的倾向最高。制剂研究数据表明乙二胺的作用机制具有双重性，且酚类化合物的存在是获得该效果的必要条件。
乙二胺以浓度依赖的方式减少了高分子量蛋白质和门冬胰岛素相关杂质的形成。
