Identification of small molecule inhibitors of the DDI2 protease

DDI2 蛋白酶小分子抑制剂的鉴定

基本信息

批准号：
10638837
负责人：
Gary L. Kleiger
金额：
$ 73.1万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-10 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10638837
关键词：
Achievement Acquired Immunodeficiency Syndrome Address Affect Apoptosis Applications Grants Area Binding Biological Assay Biophysics Cancer Patient Catalytic Domain Cell Survival Cells Cellular Assay Clinic Clinical Trials Data Detection Development Disease remission Dose Drug Targeting Dryness Enzymes Event Exhibits FDA approved Fruit Funding Future Genes Goals Grant HIV Protease Human Immune system Length Libraries Marketing Measurement Modality Multiple Myeloma Mus Normal Cell Pathway interactions Patient-Focused Outcomes Patients Peptide Hydrolases Play Powder dose form Predisposition Production Protease Inhibitor Proteasome Inhibition Proteasome Inhibitor Protein Kinase Proteins Protocols documentation Recovery Reproducibility Risk Role Signal Transduction Pathway System Testing Therapeutic Transcript Ubiquitin Up-Regulation Validation Work Xenograft Model cancer cell cancer therapy cheminformatics counterscreen expectation high throughput screening immune modulating agents inhibitor liquid chromatography mass spectrometry malignant breast neoplasm multicatalytic endopeptidase complex programs protein degradation proteostasis response scaffold scale up screening side effect small molecule inhibitor synergism targeted cancer therapy transcription factor tumor tumor growth

项目摘要

PROJECT SUMMARY Proteasome inhibitors are currently being used in the clinic against Multiple myeloma. This approach is thought to work at least in part because cancer cells appear to rely more heavily on proteasomes than do normal cells. However, our previous studies showed that proteasome inhibition invokes an adaptive program driven by the transcription factor NRF1 which upregulates proteasome genes resulting in the recovery of proteasome activity, thus limiting the efficacy of this approach. Consistent with this notion, our preliminary data suggest that depletion of NRF1 potentiates the action of proteasome inhibition therapy in a breast cancer xenograft model. Taking advantage of the fact that a protease DDI2 is essential for NRF1 activation, here we propose to identify small molecule inhibitors of this protease that could be used to enhance the efficacy of proteasome inhibitors. Moreover, based on our preliminary data that showed depleting DDI2 in itself retards tumor growth, DDI2 inhibitors could also find use as a single-agent. Based on our pilot screen that demonstrates feasibility and consistent with the goals of PAR-20-271, this 4-year project will pursue the following specific aims. In AIM 1, we will express and purify DDI2 protein and perform high-throughput screens using a protein thermal shift (PTS) assay to identify compounds that bind DDI2. In AIM 2, we will perform hit selection, confirmation and profiling using a panel of secondary assays that includes orthogonal and counter-screen assays as well as biophysical assays. In AIM 3, we will perform hit validation, hit expansion, probe selection and profile the mechanism of action of hits, followed by cellular assays to assess DDI2 target engagement, impact on the DDI2-NRF1 axis and cancer cell apoptosis.

项目概要蛋白酶体抑制剂目前在临床上用于治疗多发性骨髓瘤。这人们认为这种方法至少部分有效，因为癌细胞似乎更依赖于蛋白酶体比正常细胞更重要。然而，我们之前的研究表明，蛋白酶体抑制会调用由转录因子 NRF1 驱动的适应性程序，该程序上调蛋白酶体基因，导致蛋白酶体活性恢复，从而限制这种方法的功效。与这一观点一致，我们的初步数据表明 NRF1 的消耗增强了蛋白酶体抑制疗法在乳腺癌中的作用异种移植模型。利用蛋白酶 DDI2 对于 NRF1 至关重要的事实激活，在这里我们建议鉴定这种蛋白酶的小分子抑制剂用于增强蛋白酶体抑制剂的功效。此外，根据我们的初步数据研究表明，消耗 DDI2 本身会延缓肿瘤生长，DDI2 抑制剂也可用作单一代理人。基于我们的试点屏幕，该屏幕展示了可行性并与 PAR-20-271 的目标，这个为期 4 年的项目将追求以下具体目标。在 AIM 1 中，我们将表达和纯化 DDI2 蛋白并使用蛋白质进行高通量筛选热位移 (PTS) 测定可鉴定结合 DDI2 的化合物。在AIM 2中，我们将执行命中使用一组二次分析（包括正交分析）进行选择、确认和分析和反筛选测定以及生物物理测定。在AIM 3中，我们将执行命中接下来是验证、命中扩展、探针选择和分析命中的作用机制通过细胞测定来评估 DDI2 靶标参与情况、对 DDI2-NRF1 轴的影响以及癌细胞凋亡。