血糖仪在4种胰岛素类药物研究中的应用

目的：采用血糖仪进行德谷胰岛素和门冬胰岛素的生物学活性以及利拉鲁肽、索马鲁肽的葡萄糖耐量研究。方法：生物学活性研究：小鼠分4组，分别皮下注射标准品及高、低剂量样品40 min后，用血糖仪快速测定小鼠血糖值，第1次给药后按双交叉设计，对小鼠进行第2次给药；按照生物检定统计法中量反应平行线测定双交叉设计法计算效价及可信限率。葡萄糖耐量研究：小鼠分3组，分别皮下注射利拉鲁肽（剂量320 μg·kg^-1）、索马鲁肽（剂量320 μg·kg^-1）和空白对照，给药后24、48、72、96 h腹腔注射40%葡萄糖0.2 mL，免疫前及免疫后15、30、60 min尾静脉采血并用血糖仪测定血糖值，评价给药后小鼠葡萄糖耐量情况。结果：生物学活性研究：德谷胰岛素（CA501）制剂用3批原研德谷胰岛素为标准品，测定效价分别为97.3、108.1、91.6 U·mL^-1。德谷胰岛素（CA501）原料药和门冬胰岛素原料药效价结果分别为34.1、22.3 U·mg^-1。葡萄糖耐量研究：与空白对照相比，利拉鲁肽、索马鲁肽在给药24、48、72、96 h后注射葡萄糖15 min及30 min时仍有明显降糖作用。结论：采用血糖仪检测血糖值具有操作简便，测量快速准确，用血量少，重复性好等优点，适用于胰岛素类药物的研究。

Objective:To study the application of blood glucose meter in insulin degludec/insulin aspart biological activity assay,and in liraglutide/semaglutide glucose tolerance measurement. Methods:For biological activity assay,mice were divided into 4 groups,and after 40 minutes of immunized by high or low dose standards and samples,the blood glucose values of mice were measured by blood glucose meter immediately. A second administration were conducted as in the double cross design. The potency and fiducial limits were derived from the parallel design of the reaction volume and the design method by the bioassay statistics. For glucose tolerance measurement,the mice were divided into 3 groups,and were administrated subcutaneously with liraglutide(320 μg·kg^-1),semaglutide(320 μg·kg^-1) and blank control by injection. After 24 hours,48 hours,72 hours and 96 hours the mice were intraperitoneally injected with 40% glucose 0.2 mL,respectively. Blood glucose was measured by blood glucose meter before immunization,as well as at 15,30 and 60 minutes after immunization for glucose tolerance evaluation. Results:Using three batches of insulin degludec standard as reference,the biological activities of insulin degludec(CA501) injections were determined as 97.3,108.1 and 91.6 U·mL^-1,respectively,and that of insulin degludec(CA501) and insulin aspart active pharmaceutical ingredient was determined as 34.1 U·mg^-1,22.3 U·mg^-1,respectively. Compared with the blank control,liraglutide and semaglutide showed significant hypoglycemic efficacy even 24 hours,48 hours,72 hours and 96 hours after administration. Conclusion:Using blood glucose meter in insulin derivative biological activity assay/glucose tolerance measurement is feasible. Methods can be quick,simplified,less blood consumption with good repeatability and accuracy.

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