Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308 Tianjin, China.
Institute of Structural Biology, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health, 85764 Neuherberg, Germany.
Mol Pharm. 2020 May 4;17(5):1663-1673. doi: 10.1021/acs.molpharmaceut.0c00098. Epub 2020 Apr 13.
Glucagon-like peptide-1 (GLP-1) is an incretin (a type of metabolic hormone that stimulates a decrease in blood glucose levels), holding great potential for the treatment of type 2 diabetes mellitus (T2DM). However, its extremely short half-life of 1-2 min hampers any direct clinical application. Here, we describe the application of the heavy chain of human ferritin (HFt) nanocage as a carrier to improve the pharmacological properties of GLP-1. The GLP-HFt was designed by genetic fusion of GLP-1 to the N-terminus of HFt and was expressed in inclusion bodies in . The refolding process was developed to obtain a soluble GLP-HFt protein. The biophysical properties determined by size-exclusion chromatography (SEC), dynamic light scattering (DLS), circular dichroism (CD), transmission electron microscopy (TEM), and X-ray crystallography verified that the GLP-HFt successfully formed a 24-mer nanocage with GLP-1 displayed on the external surface of HFt. The pharmacodynamic results demonstrated that the GLP-HFt nanocage retained the bioactivity of natural GLP-1, significantly reduced the blood glucose levels for at least 24 h in a dose-dependent manner, and inhibited food intake for at least 8-10 h. The half-life of the GLP-HFt nanocage was approximately 52 h in mice after subcutaneous injection. The prolonged half-life and sustained control of blood glucose levels indicate that the GLP-HFt nanocage can be further developed for the treatment of T2DM. Meanwhile, the HFt nanocage proves its great potential as a universal carrier that improves the pharmacodynamic and pharmacokinetic properties of a wide range of therapeutic peptides and proteins.
胰高血糖素样肽-1（GLP-1）是一种肠促胰岛素（一种能降低血糖水平的代谢激素），对 2 型糖尿病（T2DM）的治疗具有巨大的潜力。然而，其半衰期极短（1-2 分钟），限制了其直接的临床应用。在这里，我们描述了利用人铁蛋白重链（HFt）纳米笼作为载体来改善 GLP-1 的药理特性的应用。通过将 GLP-1 融合到 HFt 的 N 端，设计了 GLP-HFt，并在包涵体中表达。通过复性过程开发了可溶性 GLP-HFt 蛋白。通过尺寸排阻色谱（SEC）、动态光散射（DLS）、圆二色性（CD）、透射电子显微镜（TEM）和 X 射线晶体学确定的生物物理性质证实，GLP-HFt 成功地形成了一个 24 聚体纳米笼，GLP-1 展示在 HFt 的外表面。药效学结果表明，GLP-HFt 纳米笼保留了天然 GLP-1 的生物活性，以剂量依赖的方式显著降低血糖水平至少 24 小时，并抑制进食至少 8-10 小时。皮下注射后，GLP-HFt 纳米笼在小鼠体内的半衰期约为 52 小时。半衰期延长和血糖水平持续控制表明，GLP-HFt 纳米笼可进一步开发用于治疗 T2DM。同时，HFt 纳米笼证明了其作为一种通用载体的巨大潜力，可改善广泛的治疗性肽和蛋白质的药效学和药代动力学特性。
