Small-molecule inhibitors of hADAM12

hADAM12 小分子抑制剂

基本信息

批准号：
8403416
负责人：
Jens Berthelsen
金额：
$ 2.5万
依托单位：
UNIVERSITY OF COPENHAGEN
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-31 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8403416
关键词：
ADAMTS Abate Abnormal Cell Active Sites Address Adverse effects Affinity Agreement Antineoplastic Agents Binding Biochemical Biological Assay Biological Process Biomedical Research Cancerous Catalytic Domain Cell Proliferation Cell division Cell membrane Cells Chemicals Chemistry Cleaved cell Clinical Clinical Research Clinical Trials Collaborations Collection Communities Complement Data Data Quality Data Set Development Disease Disintegrins Drosophila pros protein Drug Design Ensure Enzymes Epidermal Growth Factor Event Fluorescence Goals Growth Factor Human Development Hydroxamic Acids In Vitro Inflammatory Informatics Investigation Ions Language Libraries Malignant Neoplasms Malignant neoplasm of lung Measures Membrane Metalloproteases Metals Molecular Bank Monitor Non-Commercial Sources Penetration Peptide Hydrolases Peptides Performance Pharmaceutical Chemistry Phase Phenotype Property Provider Published Comment Research Resources Rheumatoid Arthritis Robotics Screening procedure Series Signal Transduction Solubility Specificity Structure Synthesis Chemistry TNF-alpha converting enzyme Testing Tumor Necrosis Factor-alpha Work Zinc analog assay development base cancer therapy design drug discovery extracellular high throughput screening human disease hydroxamate improved inhibitor/antagonist interest malignant breast neoplasm meetings miniaturize nervous system disorder phrases public health relevance receptor repository scaffold small molecule therapeutic target tool tumor

项目摘要

DESCRIPTION (provided by applicant): The progress of diseases often involves the dysregulation of cellular signaling events leading to altered abnormal cellular phenotypes. Cancerous diseases for instance are often characterized by an increase of abnormal cell division and proliferation due to enhanced release of growth factors such as epidermal growth factor (EGF) or tumor necrosis factor ¿ (TNF-¿). These growth factors are expressed as pro-proteins, which are recognized, cleaved and thereby activated by membrane-anchored metalloproteases (sheddases) such as a disintegrin and metalloproteases (ADAM's). Therefore, ADAMs have emerged as targets for a number of pathophysiological conditions such as cancer and neurological disorders. ADAM's are closely related to matrix metalloproteases (MMP's), which are validated targets in anti-cancer drug discovery. Although very potent MMP inhibitors have been developed, their use in cancer therapy has been hampered by a lack of efficacy and side effects in clinical trials, which are partially attributed to cross-inhibition of other zinc-dependent sheddases such as ADAM's. Thus, recent research has focused on the development of selective inhibitors addressing only the enzyme directly related to the pathophysiological condition. Most of the MMP and ADAM inhibitors in development harbor a strongly chelating hydroxamic acid (hydroxamate) group binding a zinc atom in the catalytic site, thereby blocking shedding activity. Research of such compounds has abated as they have been found to act in a relatively promiscuous manner. Using alternative chelating motifs could offer a better way of modulating binding strength and finally, binding selectivity, which would significantly improve the usefulness of metalloprotease inhibitors in biochemical and biomedical research as well as cancer therapy. While ADAM17, also known as TNF-¿ converting enzyme (TACE), is probably the most prominent ADAM explored as a target for the treatment of lung and breast cancer and inflammatory diseases such as rheumatoid arthritis, other ADAM's such as ADAM12 are much less characterized potential targets for therapeutic purposes. Therefore, the specific aim of this proposal is to screen the Molecular Libraries Small Molecule Repository (MLSMR) for ADAM12 inhibitors focusing on the discovery of compounds with chelating motifs other than the traditional hydroxamate group in the downstream secondary screening and medicinal chemistry efforts in order to minimize cross-inhibition of other ADAM's. A selectivity panel of five ADAM proteases related to ADAM12 will allow to assess binding selectivity in both biochemical and cell-based assays monitoring the shedding of natural ADAM substrates, thereby generating a comprehensive data set on the inhibitory profile of biomedically interesting ADAM proteases. The primary high-throughput screening will employ a fluorescence-based assay measuring the cleavage of a quenched substrate peptide. This assay has recently been miniaturized to 1536- well plate format demonstrating the robustness required for an unattended robotic screening of the MLSMR compound library and a sufficient performance to ensure data quality for subsequent analysis. Cross-validated compounds derived after primary and secondary assays will be further developed to enhance potency and selectivity for ADAM12. !

描述（由申请人提供）：疾病的进展通常涉及细胞信号传导事件的失调，从而导致异常细胞表型的改变，例如，癌症疾病的特征通常是由于生长因子（例如生长因子）的释放增加而导致异常细胞分裂和增殖的增加。作为表皮生长因子 (EGF) 或肿瘤坏死因子 ¿这些生长因子以前蛋白的形式表达，可被膜锚定的金属蛋白酶（脱落酶）如解整合素和金属蛋白酶 (ADAM) 识别、切割并激活。因此，ADAM 已成为治疗的靶点。许多病理生理学病症，例如癌症和神经系统疾病，与基质金属蛋白酶 (MMP) 密切相关。尽管已经开发出非常有效的 MMP 抑制剂，但它们在癌症治疗中的应用却因临床试验中缺乏疗效和副作用而受到阻碍，部分原因是与其他锌离子的交叉抑制。因此，最近的研究集中在开发仅针对与病理生理状况直接相关的酶的强选择性抑制剂，大多数正在开发的 MMP 和 ADAM 抑制剂都含有螯合异羟肟酸。（异羟肟酸）基团在催化位点结合锌原子，从而阻止脱落活性的研究已经减少，因为人们发现它们以相对混杂的方式发挥作用，可以提供更好的调节结合强度的方法。最后，结合选择性，这将显着提高金属蛋白酶抑制剂在生化和生物医学研究以及癌症治疗中的用途。转化酶 (TACE) 可能是作为治疗肺癌和乳腺癌以及类风湿性关节炎等炎症性疾病的最重要的 ADAM 靶点，而 ADAM12 等其他 ADAM 的潜在治疗靶点特征较少。该提案的具体目的是在分子库小分子库 (MLSMR) 中筛选 ADAM12 抑制剂，重点是发现具有除传统异羟肟酸基团之外的螯合基序的化合物下游二次筛选和药物化学工作，以最大限度地减少其他 ADAM 的交叉抑制。与 ADAM12 相关的五种 ADAM 蛋白酶的选择性小组将能够评估监测天然 ADAM 底物脱落的生化和基于细胞的测定中的结合选择性。，从而生成生物医学上感兴趣的 ADAM 蛋白酶的抑制谱的综合数据集。初级高通量筛选将采用基于荧光的测定来测量 a 的裂解。该测定最近已小型化为 1536 孔板格式，证明了无人值守的 MLSMR 化合物库机器人筛选所需的稳健性，以及确保初次和交叉验证后得出的化合物的数据质量的足够性能。将进一步开发二次测定以增强 ADAM12 的效力和选择性！