Characterization of small molecule inhibitors of HDACs and DNMTs

HDAC 和 DNMT 小分子抑制剂的表征

基本信息

批准号：
7933340
负责人：
ELISABETH D MARTINEZ
金额：
$ 6.05万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2010-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7933340
关键词：
Acetylation Affect Automation Biological Biological Assay Breast Cancer Cell Cancer Biology Cancer Cell Growth Cancer cell line Cell Line Cells Cellular Assay Chemicals Chimeric Proteins Chromatin Clinical Trials DNA DNA Methyltransferase DNA Methyltransferase Inhibitor DNA Modification Methylases Development Dose Drug Evaluation Enzymes Epigenetic Process Evaluation Event Family Fatty acid glycerol esters Fluorescence Foundations Funding Gene Activation Gene Expression Gene Silencing Gene Targeting Genes Goals Growth Histone Deacetylase Inhibitor Histones Human Image In Vitro Inhibitory Concentration 50 Lead Luciferases Lung Malignant Neoplasms Malignant neoplasm of lung Mammalian Cell Mammary gland Measures Mediating Methods Methylation Modification Monitor Mus Nude Mice Pathway interactions Pharmaceutical Preparations Phase Process Promoter Regions Property Proteins Protocols documentation Reporter Genes Research Personnel Reverse Transcriptase Polymerase Chain Reaction System Testing Therapeutic Transcriptional Regulation Transferase Tumor Suppressor Genes Work Xenograft Model Xenograft procedure analog base cancer cell cancer therapy cell growth drug candidate drug sensitivity enzyme activity enzyme pathway gene repression improved in vivo inhibitor/antagonist innovation neoplastic cell novel pre-clinical programs promoter protein function public health relevance small molecule small molecule libraries transcription factor tumor tumor growth tumorigenic

项目摘要

DESCRIPTION (provided by applicant): Over the last several years, a few drugs have been identified that inhibit the methylation or de-acetylation pathways mediated by DNA methyltransferases (DNMT) and histone deacetylases (HDAC). These compounds have had immediate application in the treatment of cancers because of their ability to reactivate aberrantly silenced tumor suppressor genes. The need to identify more small molecules that target these two enzyme families is clear and has been repeatedly stated in the field of cancer biology. Our overall goal is to expand the available repertoire of drugs that can modulate gene expression and to evaluate their basic mechanism of action. Toward this goal, we have recently developed a unique mammalian cell-based system to screen for such small molecules in a reliable, simple fashion that is amenable to automation. Unlike methods used to date, our system has the advantage of measuring the exact biological event it seeks to target: the reversal of transcriptional repression in mammalian cells. The assay has been tested in screens of small molecule libraries and has successfully resulted in eight hits. Preliminary evaluation of one hit compound and one analog has revealed HDAC inhibitor properties. Because these compounds are structurally distinct from known HDAC/DNMT inhibitors, they will contribute to improving the diversity of this drug type. The funding of this proposal will allow us to characterize the mechanism of action of the identified compounds and to evaluate their anti-cancer activity. Our specific aims are: 1: To measure the ability of the compounds identified in our high content screen to inhibit the enzymatic activity of HDAC or DNMT enzymes in vitro; 2: To evaluate the ability of the candidate epigenetic modulators to inhibit the growth of human cancer cells and to re-activate the expression of aberrantly silenced genes in human cancer cell lines; and 3: To measure the effects of these epigenetic modulators on tumor growth in xenograft models. These studies will provide the foundation for further work in the development of these potential cancer therapeutic compounds and allow us to move into pre-clinical drug evaluation. Public health relevance: We have developed a system to identify drugs with the ability to turn on genes that are abnormally turned off in cancer cells. Using this system, we have already identified eight candidate drugs and propose to characterize them measuring their ability to block the growth of human cancer cells.

描述（由申请人提供）：在过去的几年中，已经发现了一些药物抑制由DNA甲基转移酶（DNMT）和组蛋白脱乙酰基酶（HDAC）介导的甲基化或去乙酰化途径。这些化合物在癌症的治疗中立即应用，因为它们能够重新激活抑制肿瘤基因的异常沉默。鉴定靶向这两个酶家族的更多小分子的需求是明显的，并且在癌症生物学领域反复说明。我们的总体目标是扩大可以调节基因表达并评估其基本作用机理的药物的可用曲目。为了实现这一目标，我们最近开发了一种独特的基于哺乳动物的细胞系统，以可靠，简单的方式筛选出如此小的分子，该分子可以自动化。与迄今为止使用的方法不同，我们的系统具有测量其试图靶向的确切生物事件的优点：哺乳动物细胞中转录抑制的逆转。该测定已在小分子库的屏幕上进行了测试，并成功导致了八次命中。对一种HIT化合物和一种类似物的初步评估揭示了HDAC抑制剂特性。由于这些化合物在结构上与已知的HDAC/DNMT抑制剂不同，因此它们将有助于改善该药物的多样性。该提案的资金将使我们能够表征已确定的化合物的作用机理，并评估其抗癌活性。我们的具体目的是：1：测量在我们的高含量筛选中鉴定的化合物在体外抑制HDAC或DNMT酶的酶活性的能力； 2：评估候选表观遗传调节剂抑制人类癌细胞生长的能力，并重新激活人类癌细胞系中异常沉默的基因的表达；和3：测量这些表观遗传调节剂对异种移植模型中肿瘤生长的影响。这些研究将为进一步开发这些潜在的癌症治疗化合物的发展提供基础，并使我们能够进行临床前的药物评估。公共卫生相关性：我们已经开发了一种系统来识别具有打开癌细胞异常关闭基因的药物。使用该系统，我们已经确定了八种候选药物，并提出了测量其阻断人类癌细胞生长的能力的表征。