FPRL1-specific anti-inflammatory compounds: Alzheimer's

FPRL1 特异性抗炎化合物：阿尔茨海默病

• 批准号: 7107339
• 负责人: JESSE BAUMGOLD
• 金额: $ 10.6万
• 依托单位: APPLIED CELL SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7107339
• 关键词: Alzheimer' s disease; CHO cells; amyloid proteins; antiinflammatory agents; bioaccumulation; calcium flux; cell line; cell surface receptors; chemical registry /resource; drug design /synthesis /production; guanine nucleotide binding protein; inhibitor /antagonist; neural degeneration; neuritic plaques; protein structure function; radiotracer; receptor expression; western blottings

DESCRIPTION (provided by applicant): The overall objective of this project is to develop an antagonist for the G-protein-coupled receptor FPRL1 (formyl peptide receptor-like 1) to test the hypothesis that blocking this receptor will prevent accumulation of inflammatory cells into plaques in Alzheimers disease, and thereby prevent neuronal degeneration. FPRL1 receptors have been shown to be expressed on human mononuclear phagocytes which accumulate in plaques and tangles in AD brain. Further, the 42-amino acid form of beta-amyloid has been shown to activate FPRL1 receptors and induce chemotaxis in these phagocytes. Thus, by blocking the FPRL1 receptor we expect to prevent accumulation of these inflammatory cells in AD lesions and consequent neuronal degeneration resulting from inflammatory processes. In this Phase I application, we first propose constructing and validating a cell line that expresses both the FPRL1 receptor and a promiscous G protein that will allow this receptor to couple to the intracellular calcium pathway. This cell line will allow us to rapidly and inexpensively screen a highly diverse library of 14,000 compounds, for antagonist activicty at the FPRL1 receptor. This library includes novel and highly diverse compounds from 50 different structural classes that have rigid scaffold and are amenable to rapid synthetic modification for later lead optimization. Our screening criteria is to find at least one compound that blocks the FPRL1 -mediated increase in intracellular calcium by 50% or more at a concentration of 10 uM. Compounds meeting this criteria will be evaluated further in the following assays: 1) full EC50 curves will be obtained for such compounds in the same intracellular calcium assay; 2) inhibition of peptide W mediated increase in GTP-g-S binding to membranes from cells expressing the FPRL1 receptor; 3) inhibition of radio-iodinated peptide W binding to membranes from cells expressing FPRL1 receptor and 4) inhibition of amyloid beta 42 mediated increase in intracellular calcium in whole cells. Compounds having Ki values of 10 uM or lower in all of these assays (except the peptide W binding assay) will be carried forward to a Phase 2 application in which they will be optimized and rendered potent at the nanomolar level. In Phase II, optimized compounds will be tested in a mouse model of AD for their ability to prevent phagocyte accumulation into brain lesions. Ultimately, the best compound will be licensed to a pharmaceutical company for clinical trials.

描述（由申请人提供）：该项目的总体目的是为G蛋白偶联受体FPRL1（甲基肽受体样1）开发拮抗剂，以检验以下假设：阻断该受体将防止炎性细胞积累到阿尔茨海默氏病中的斑块中，并在阿尔茨海默氏病中及其预防神经元。 FPRL1受体已被证明是在人类单核吞噬细胞上表达的，该吞噬细胞积聚在AD脑中的斑块和缠结中。此外，已经显示出42个氨基酸形式的β-淀粉样蛋白会激活FPRL1受体并在这些吞噬细胞中诱导趋化性。因此，通过阻断FPRL1受体，我们期望阻止这些炎症细胞在AD病变中的积累，并导致炎症过程导致的神经元变性。在此阶段I应用中，我们首先提出构建和验证一个表达FPRL1受体和渗透G蛋白的细胞系，该细胞系将使该受体将其搭配到细胞内钙途径。该细胞系将使我们能够快速且廉价地筛选一个高度多样化的14,000种化合物的文库，以获取FPRL1受体的拮抗剂activicty。该库包括来自50种不同结构类别的新颖和高度多样的化合物，这些化合物具有刚性脚手架，并且可以随时进行合成的修饰，以便以后的铅优化。我们的筛选标准是找到至少一种化合物，该化合物以10 um的浓度阻止了FPRL1介导的细胞内钙的增加50％或更多。符合此标准的化合物将在以下测定中进一步评估：1）在同一细胞内钙测定中，将获得此类化合物的完整EC50曲线； 2）抑制肽W介导的GTP-G-S与表达FPRL1受体的细胞结合的GTP-G-S结合； 3）抑制来自表达FPRL1受体的细胞和4）淀粉样蛋白β的细胞与膜的抑制与膜的结合。42整个细胞中细胞内钙的介导的增加。在所有这些测定中（肽W结合测定法除外）的Ki值为10 um或较低的化合物将被带到2阶段应用中，在该应用中，将在纳莫尔级别进行优化并在纳摩尔水平上实现有效性。在II期中，将在AD的小鼠模型中测试优化的化合物，以防止吞噬细胞积累到脑损伤中。最终，最好的化合物将获得制药公司的临床试验许可。