Laboratory for Pharmaceutical Biotechnology - pbiotech. Federal University of Rio de Janeiro - UFRJ, CCS, Bss24, Ilha do Fundão, 21941-590 Rio de Janeiro, RJ, Brazil.
Laboratory for Pharmaceutical Biotechnology - pbiotech. Federal University of Rio de Janeiro - UFRJ, CCS, Bss24, Ilha do Fundão, 21941-590 Rio de Janeiro, RJ, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB-INCT), Federal University of Rio de Janeiro, Rio de Janeiro 21941-590, Brazil.
Int J Pharm. 2018 Aug 25;547(1-2):621-629. doi: 10.1016/j.ijpharm.2018.06.039. Epub 2018 Jun 19.
Since the discovery of amylin its use has been discouraged by the inadequacy of the protocol involving multiple injections in addition to insulin. We aimed here to develop a combined fixed-dose formulation of pramlintide with fast-acting insulin. We have investigated the compatibility of regular and fast-acting insulin analogues (Aspart, Asp, and LisPro, LysPro) with the amylin analogue pramlintide by using electrospray ionization - ion mobility spectrometry-mass spectrometry (ESI-IMS-MS), kinetic aggregation assays monitored by thioflavin T, and transmission electron microscopy (TEM) in the evaluation of the aggregation product. Insulin interacts with pramlintide, forming heterodimers as probed by ESI-IMS-MS. While their interaction is likely to delay the amyloid aggregation of pramlintide in phosphate-buffered solution pH 7.0, they do not prevent aggregation at this condition. At acidic sodium acetate solution pH 5.0, combination of pramlintide and the fast-acting insulin analogues become stable against amyloid aggregation. The co-formulated product at high concentration of both pramlintide (600 μg/mL,150 μM) and LisPro insulin (50 IU/mL, 300 μM) showed also stability against amyloid aggregation. These data indicate the physico-chemical short-term stability of the co-formulated preparation of LisPro insulin with pramlintide, which could bring benefits for the combined therapy.
自发现胰淀素以来，由于涉及胰岛素多次注射的方案不够完善，其应用受到了阻碍。我们旨在开发一种与速效胰岛素联合使用的普兰林肽固定剂量制剂。我们使用电喷雾电离-离子淌度质谱（ESI-IMS-MS）、用硫代黄素 T 监测的动力学聚集测定法以及透射电子显微镜（TEM）研究了常规和速效胰岛素类似物（Aspart、Asp 和 LisPro、LysPro）与胰淀素类似物普兰林肽的相容性，以评估聚集产物。胰岛素与普兰林肽相互作用，通过 ESI-IMS-MS 探测形成异二聚体。虽然它们的相互作用可能会延迟磷酸缓冲溶液 pH7.0 中普兰林肽的淀粉样聚集，但在这种条件下并不能阻止聚集。在酸性乙酸钠溶液 pH5.0 下，普兰林肽和速效胰岛素类似物的组合变得稳定，不易发生淀粉样聚集。在高浓度的普兰林肽（600μg/mL，150μM）和 LisPro 胰岛素（50IU/mL，300μM）的共同配方产品也显示出对淀粉样聚集的稳定性。这些数据表明 LisPro 胰岛素与普兰林肽联合制剂的物理化学短期稳定性，这可能为联合治疗带来益处。
