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    • 8. 发明申请
    • PROTEIN AGENT FOR DIABETES TREATMENT AND BETA CELL IMAGING
    • 蛋白质类药物治疗糖尿病和β细胞成像
    • WO2011050052A3
    • 2011-09-15
    • PCT/US2010053361
    • 2010-10-20
    • UNIV GEORGIA STATE RES FOUNDLIU ZHI-RENYANG JIEXU BINGZHOU WANGDAXUE SHENGUI
    • LIU ZHI-RENYANG JIEXU BINGZHOU WANGDAXUE SHENGUI
    • C07K19/00A61K38/26A61K49/14A61P3/10G01N33/52
    • A61K38/26A61K38/00A61K47/60A61K49/085C07K14/605C07K2319/00
    • Glucagon-like peptide-1 (GLP-1 ) is a member of a large family of incretin hormones secreted in nutrient-dependent response. GLP-1 acts on GLP-1 receptor (GLP-1 R) that is highly expressed on pancreatic ß-cells. The peptide has great potential for development of diabetes treatment and diagnosis. However, the pharmaceutical effects of the peptide suffer from in vivo instability and short life due to degradation by Dipeptidylpeptidase-1 (DPP-4). The 30 amino acid peptide GLP-1 has been integrated into a stable host protein human calbindin D9k. The fusion protein binds to GLP-1 R as demonstrated by immunostaining analyses of GLP-1 R expressing cells. The fusion protein agents can be useful for both diabetes treatment and GLP-1 R receptor targeting MR imaging. The fusion protein has a size about 14 kDa, which enables efficient tissue penetration and retention, and an extended circulation time, is stable, remaining intact and retaining activity after 48 hours incubation with 75% human serum. The protein retains its native folded structure after boiling for ten minutes forming the basis of an experimental protocol for large scale production of the fusion protein (>30 mg/l bacterial culture). No toxicity has been observed with tests on mice. One aspect of the disclosure, therefore, provides a fusion protein comprising a first peptide characterized by selectively binding to a site of a target cell and linked to a second peptide, where the fusion protein is more stable than the first peptide alone. In embodiments of this aspect of the disclosure, the fusion protein may further comprise a detectable label attached thereto. In some embodiments of this aspect of the disclosure, the first peptide of the fusion protein may be glucagon-like peptide-1 (GLP-1 ), glucagon-like peptide-1 (GLP-1 )(7-36), or glucagon-like peptide-1 (GLP-1 ) (9-36), or a conservative variant thereof.
    • 胰高血糖素样肽-1(GLP-1)是营养物依赖性反应中分泌的肠降血糖素激素大家族的成员。 GLP-1作用于在胰岛β细胞上高表达的GLP-1受体(GLP-1R)。 该肽具有发展糖尿病治疗和诊断的巨大潜力。 然而,由于二肽基肽酶-1(DPP-4)的降解,肽的药物作用受到体内不稳定性和短寿命的影响。 已将30个氨基酸的肽GLP-1整合到稳定的宿主蛋白人类钙结合蛋白D9k中。 如通过表达GLP-1R的细胞的免疫染色分析所证实的,融合蛋白与GLP-1R结合。 融合蛋白剂可用于糖尿病治疗和GLP-1R受体靶向MR成像。 该融合蛋白具有约14kDa的大小,其使得有效的组织穿透和保留以及延长的循环时间是稳定的,在用75%人血清孵育48小时后保持完整并保持活性。 沸腾10分钟后,蛋白质保留其天然折叠结构,形成大规模生产融合蛋白(> 30mg / l细菌培养物)的实验方案的基础。 对小鼠的测试没有观察到毒性。 因此,本公开的一个方面提供了包含第一肽的融合蛋白,所述第一肽的特征在于选择性结合靶细胞的位点并与第二肽连接,其中融合蛋白比单独的第一肽更稳定。 在本公开内容的该方面的实施方案中,融合蛋白可以进一步包含与其连接的可检测标记。 在本公开的该方面的一些实施方案中,融合蛋白的第一肽可以是胰高血糖素样肽-1(GLP-1),胰高血糖素样肽-1(GLP-1)(7-36)或胰高血糖素 (GLP-1)(9-36)或其保守变体。