A liquid chromatography high-resolution mass spectrometry in vitro assay to assess metabolism at the injection site of subcutaneously administered therapeutic peptides
A liquid chromatography high-resolution mass spectrometry in vitro assay to assess metabolism at the injection site of subcutaneously administered therapeutic peptides
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DOI：
10.1016/j.jpba.2018.07.009
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发表时间：
2018-09-10
影响因子：
3.4
通讯作者：
Monteagudo, Edith
中科院分区：
文献类型：
--
作者：
Esposito, Simone;de Leonibus, Maria Lucia;Monteagudo, Edith
皮下（SC）注射是肽治疗药物最常见的给药途径。注射部位的分解代谢可能是许多皮下注射肽的特定且主要的降解途径。在某些情况下，它会显着影响药代动力学，特别是生物利用度，并对药物的功效产生不利影响。这项工作描述了一种基于液相色谱-高分辨率质谱的体外测定法，用于评估 SC 组织中的肽代谢（SCiMetPep 测定）。 SCiMetPep 测定是使用人、Sprague-Dawley 大鼠和哥廷根小型猪 SC 组织匀浆上清液开发的，可测定母肽的降解率（半衰期）并鉴定酶促蛋白水解产生的代谢物。该测定是使用已知肽（包括人胰岛素和四种 GLP-1 类似物（利西那肽、艾塞那肽、利拉鲁肽和索马鲁肽））开发和验证的。为了优化 SC 组织的均质过程和肽孵育条件，评估了不同的实验参数。这些肽的体外代谢与文献报道的体内数据非常一致。最后，当 SCiMetPep 检测应用于一系列结构相关的肽时，发现 SC 代谢稳定性和生物利用度之间存在相当好的相关性，这表明注射部位的分解代谢在肽治疗药物的吸收、分布、代谢和排泄 (ADME) 中发挥着重要作用。 SCiMetPep 显示出能够识别具有改善的 SC 代谢稳定性并因此具有更高生物利用度的类似物。该测定可用于药物发现的早期阶段，以识别注射部位的肽代谢软点并指导肽药物发现过程。 (C) 2018 Elsevier B.V. 保留所有权利。
Subcutaneous (SC) injection is the most common administration route for peptide therapeutics. Catabolism at the injection site can be a specific and major degradation pathway for many SC administered peptides. In some cases, it can significantly affect pharmacokinetics, particularly bioavailability, and have detrimental effects on the efficacy of the drug.This work describes a liquid chromatography-high resolution mass spectrometry based in vitro assay to assess peptide metabolism in the SC tissue (SCiMetPep assay). The SCiMetPep assay was developed using human, Sprague-Dawley rat and Gottingen minipig SC tissue homogenate supernatant, and allows for both determination of degradation rate (half-life) of the parent peptide and identification of metabolites generated from enzymatic proteolysis.The assay was developed and validated using known peptides including human insulin and four GLP-1 analogues (lixisenatide, exenatide, liraglutide and semaglutide). Different experimental parameters were evaluated in order to optimize the homogenization process of the SC tissue and the peptide incubation conditions. In vitro metabolism of these peptides was in good agreement with in vivo data reported in the literature. Finally, when SCiMetPep assay was applied on a series of structurally related peptides, a fairly good correlation was found between SC metabolic stability and bioavailability, suggesting that catabolism at the injection site can have a major role in the absorption, distribution, metabolism, and excretion (ADME) of peptide therapeutics. The SCiMetPep showed the ability to identify analogs with improved SC metabolic stability and hence higher bioavailability. The assay can be used in the early phases of drug discovery to identify peptide metabolic soft spots at the injection site and guide the peptide drug discovery process. (C) 2018 Elsevier B.V. All rights reserved.