The effect of absorption enhancers on the initial degradation kinetics of insulin by α-chymotrypsin
The effect of absorption enhancers on the initial degradation kinetics of insulin by α-chymotrypsin
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DOI：
10.1016/s0378-5173(01)00595-6
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发表时间：
2001-04-17
影响因子：
5.8
通讯作者：
Aboul-Enein, HY
中科院分区：
文献类型：
--
作者：
Radwan, MA;Aboul-Enein, HY
本研究的目的是建立一种快速筛选各种胰岛素吸收促进剂的方法，通过跟踪它们对胰岛素与α-糜蛋白酶在37 ℃孵育的初始动力学的影响。建立了一种简便、灵敏、重现性好的反相高效液相色谱法。在0.4 - 4 U/ml胰岛素范围内观察到线性响应（r> 0.999）。日间和日内胰岛素研究之间无显著性差异（P <0.05）。日内精密度和准确度测定结果的平均相对标准偏差（RSD %）均<12%。由于添加了任何吸收促进剂，该试验对检测任何代谢产物的存在具有灵敏度，即使在单独使用胰岛素时未观察到。仅当胰岛素与α-胰凝乳蛋白酶在37 ℃孵育时才检测到三种代谢物（A-C）。当甘氨胆酸（0.5，1%）或牛磺鹅去氧胆酸盐（0.5，1%）与胰岛素在不存在α-胰凝乳蛋白酶的情况下于37 ℃孵育时，观察到代谢产物D。显著增加胰岛素T-50%的化合物为胆酸（0.5%）>脱氧胆酸（1%）>脱氧胆酸（0.5%）>胆酸（1%）>胆酸（0.5，1%）。癸酸（0.5%）、羟丙基-α-环糊精（0.5，1%）和二甲基-α-环糊精（0.5，1，5%）对胰岛素T-50%无显著影响。胆汁盐按以下顺序增加胰岛素T-50%：脱氧胆酸盐>胆酸盐>甘氨胆酸盐>牛磺胆酸盐>牛磺脱氧胆酸盐>牛磺鹅脱氧胆酸盐>甘氨脱氧胆酸盐。所获得的结果将支持利用这种方法筛选掺入胰岛素制剂中的任何化合物的可行性。这些化合物应该以尽可能低的浓度使用，以避免或最小化胰岛素降解。(C)2001 Elsevier Science B.V.保留所有权利。
The goal of this investigation was to establish a fast method to screen various insulin absorption enhancers by following their effect on the initial kinetics of insulin incubated with alpha -chymotrypsin at 37 degreesC. A simple, sensitive and reproducible reversed phase high performance liquid chromatography (HPLC) method has been developed to carry out this goal. Linear responses (r > 0.999) were observed over the range of 0.4-4 U/ml for insulin. There was no significant difference (P < 0.05) between inter- and intra-day studies for insulin. The mean relative standard deviations (RSD%) of the results of within-day precision and accuracy of insulin were < 12%. The assay was sensitive to detect the existence of any metabolite due to the addition of any absorption enhancers, even if it was not seen with insulin alone. Three metabolites (A-C) were detected only when insulin was incubated with alpha -chymotrypsin at 37 degreesC. Metabolite D was observed when either glycocholic acid (0.5, 1%) or taurochenodeoxycholate (0.5, 1%) was incubated with insulin in the absence of alpha -chymotrypsin at 37 degreesC. The compounds that significantly increased insulin T-50% were glycyrrhizic acid (0.5%) > deoxycholic acid (1%) > deoxycholic acid (0.5%) > glycyrrhizic acid (1%) > cholic acid (0.5, 1%). Capric acid (0.5%), hydroxypropyl-alpha -cyclodextrin (0.5, 1%) and dimethyl-alpha -cyclodextrin (0.5, 1, 5%) did not significantly affect insulin T-50%. The bile salts increased insulin T-50% in this order: deoxycholate > cholate > glycocholate > taurocholate > taurodeoxycholate > taurochenodeoxycholate > glycodeoxycholate. The results obtained would support the feasibility of utilizing such method for screening any compound incorporated in insulin formulation. These compounds should be used in the minimum possible concentration to avoid or minimize insulin degradation. (C) 2001 Elsevier Science B.V. All rights reserved.