Multiplex cell-based platform for kinase drug discovery

用于激酶药物发现的多重细胞平台

• 批准号: 8394905
• 负责人: Peter Krutzik
• 金额: $ 18.84万
• 依托单位: PRIMITY, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-18 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8394905
• 关键词: Address; Antigens; Antineoplastic Agents; Apoptosis; Biochemical; Biological Assay; Cell Line; Cell membrane; Cells; Clinical Trials; Complex; Coupled; Development; Dimerization; Disease; Drug Delivery Systems; Environment; Exhibits; FDA approved; Flow Cytometry; Generations; Grant; Growth; Hour; Human Genome; In Vitro; Individual; Induction of Apoptosis; Interleukin-3; Lead; Libraries; Malignant Neoplasms; Measurement; Measures; Neurologic; Oncogenic; Pathogenesis; Pattern; Pharmaceutical Preparations; Phosphotransferases; Population; Preclinical Drug Evaluation; Protein Tyrosine Kinase; Reading; Receptor Protein-Tyrosine Kinases; Reporting; Research Personnel; Screening procedure; Signal Pathway; Staging; System; Technology; Time; Toxic effect; Tube; Tyrosine; Tyrosine Kinase Inhibitor; Validation; Vendor; base; cancer therapy; cost; density; design and construction; drug discovery; effective therapy; high throughput screening; improved; inhibitor/antagonist; kinase inhibitor; mutant; novel; oncology; programs; response; small molecule; stability testing; success; tool; vector

DESCRIPTION (provided by applicant): There are 518 kinases in the human genome, of which at least 30 have been targeted in clinical trials for different diseases. Most of these targeted kinases are tyrosine kinases, and particularly receptor tyrosine kinases, which are targeted by over half of all FDA, approved kinase inhibitors. In many cancers, drugs that target a single kinase often lack efficacy due to the presence of alternate oncogenic signaling pathways that may compensate for the inhibition of the targeted kinase, or mutant versions of the targeted kinase that are unaffected by the candidate compound. Although broadly non-selective kinase inhibitors are highly toxic, it has been shown that selective and simultaneous inhibition of a number of kinases can lead to more effective cancer therapies. Thus, one of the major challenges facing kinase drug discovery is how to efficiently create second generation inhibitors that block multiple selected kinases while avoiding toxicity associated with broad spectrum activity. Although current in vitro biochemical assays are available to measure selectivity of compounds against most of the kinome, these assays are cost-prohibitive to perform on a large scale, and do not adequately replicate the complex cellular environment of the target kinases. Current cell-based assays are usually limited to measuring one kinase per assay well and therefore are not amenable to rapid screening for multipotent inhibitors. Here we propose the creation of a second generation cell-based kinase screening tool that measures the effects of compounds across dozens of kinases in a single well. The ability to simultaneously screen multiple targets will promote the discovery of selectively non-selective compounds that promise more effective therapies. The proposed assay platform takes advantage of Primity's CellCode multiplexing technology which encodes cell populations with unique fluorescent signatures such that multiple cell lines can be combined in a single tube. This technology coupled with the adaptation and optimization of an established, universal, cell-based assay for discovering tyrosine kinase inhibitors enables detailed selectivity measurements and compound screening against the tyrosine kinome at a scale that was previously not possible. PUBLIC HEALTH RELEVANCE: Kinases are implicated in the development and pathogenesis of a wide array of diseases ranging from cancer to neurological and immunological disorders. Here we describe a platform that determines the effects of kinase inhibitors across dozens of targets simultaneously to discover novel drugs with unique patterns of inhibition and to understand potential toxicities of current therapies. This system will improve kinase drug screening and accelerate discovery of novel cancer treatments.

描述(申请人提供)：人类基因组中有518个激酶，其中至少有30个已被用于不同疾病的临床试验。这些靶向的激酶大多是酪氨酸激酶，特别是受体酪氨酸激酶，超过一半的FDA批准的激酶抑制剂都以受体酪氨酸激酶为靶点。在许多癌症中，靶向单一激酶的药物往往缺乏疗效，这是因为存在替代的致癌信号通路，可以补偿靶向激酶的抑制，或者靶向激酶的突变版本不受候选化合物的影响。尽管广泛的非选择性激酶抑制剂毒性很高，但已有研究表明，选择性地同时抑制一些激酶可以导致更有效的癌症治疗。因此，激酶药物开发面临的主要挑战之一是如何有效地创造第二代抑制剂来阻断多个选定的激酶，同时避免与广谱活性相关的毒性。虽然目前的体外生化分析可以用来测量化合物对大多数亲缘关系的选择性，但这些分析成本高昂，不能大规模进行，并且不能充分复制目标激酶的复杂细胞环境。目前基于细胞的检测通常仅限于每次检测一种激酶，因此不适用于快速筛选多能抑制剂。在这里，我们建议创建一种第二代基于细胞的激酶筛选工具，该工具可以测量单个井中数十个激酶的化合物的影响。同时筛选多个靶点的能力将促进选择性非选择性化合物的发现，这些化合物有望提供更有效的治疗。建议的分析平台利用Primity的CellCode多路传输技术，该技术使用独特的荧光信号对细胞群进行编码，从而可以将多个细胞系组合在一个试管中。这项技术再加上发现酪氨酸激酶抑制剂的成熟的、通用的、基于细胞的分析的适应和优化，使得能够以以前不可能的规模进行详细的选择性测量和针对酪氨酸激酶的化合物筛选。 与公共卫生相关：从癌症到神经和免疫疾病，一系列疾病的发生和发病都与激酶有关。在这里，我们描述了一个平台，它可以同时确定激酶抑制剂在数十个靶点上的效果，以发现具有独特抑制模式的新药，并了解当前治疗方法的潜在毒性。该系统将改进激酶药物的筛选，并加速发现新的癌症治疗方法。