Cell-free assay technologies for the identification of active compounds

用于鉴定活性化合物的无细胞测定技术

基本信息

批准号：
10926223
负责人：
Barry Okeefe
金额：
$ 68.34万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

项目摘要

The PCMBS collaborates closely with the MTP Assay Development and Screening Section (ADSS/Henrich) to automate, validate and complete high throughput screens (HTS), with the Chemical Diversity and Development Section (CDDS/Beutler) to access new sources of chemical diversity and with the Natural Products Chemistry Section (NPCS/Grkovic, Du) to prioritize extracts for natural product isolation and biochemically evaluate pure active compounds isolated by NPCS or CDDS. Examples of PCMBS efforts during this review period, include published papers on screens of natural products for selective inhibitors of the protease MALT the ubiquitin ligase CBLB, and the phosphodiesterase TDP1. Our studies on natural product inhibitors of TDP1 also resulted in a second paper describing a new class of cyclic peptides which are the first reported allosteric inhibitors of this phosphodiesterase. We have recently also completed a screen for inhibitors of the phosphodiesterase TDP2 with a manuscript in preparation. In addition, the PCMBS has continued to increase the implementation of biophysical screening methods for the identification of specific modulators of the thermal stability of both protein and RNA structures. Initially, this work resulted in the identification of a new small molecule scaffold that altered the stability of HIV-1 TAR RNA. This technology has now been expanded upon to allow for the facile screening of pure compounds and natural product samples in a high throughput format to identify modulators of RNA and protein stability. One recently published example was the identification of a natural product that selectively modulated the thermal stability of pre-miR-21. We have recently completed two new enzymatic screens, against the chimeric kinase DNAJB1-PRKACA (PKADJ) and inositol 1,3,4-triphosphate 5/6 kinase (ITPK1), as well as four biophysical screens; against the immunologically-pertinent metabolic enzyme Immune Responsive Gene 1 (IRG1), the diagnostic target for hepatocellular carcinoma glypican-3 (GPC-3) and the SARS-CoV-2 receptor binding domain (RBD), and DNA-reading protein BRD4. The PCMBS also evaluates compounds derived from MTP screens for their specific interactions against macromolecular targets. We have used both biophysical and biochemical techniques to evaluate inhibitors of pre-miR-21 RNA, TDP1, MALT1,CBLB, and SARS-CoV-2 viral entry.

The PCMBS collaborates closely with the MTP Assay Development and Screening Section (ADSS/Henrich) to automate, validate and complete high throughput screens (HTS), with the Chemical Diversity and Development Section (CDDS/Beutler) to access new sources of chemical diversity and with the Natural Products Chemistry Section (NPCS/Grkovic, Du) to prioritize extracts for natural product isolation and biochemically evaluate pure active NPC或CDD分离的化合物。在此审查期间，PCMBS努力的示例包括有关天然产品筛选的发表论文，用于蛋白酶麦芽蛋白酶泛素连接酶CBLB和磷酸二酯酶TDP1的选择性抑制剂。我们对TDP1天然产物抑制剂的研究也导致了第二篇论文，描述了新的环状肽，这是该磷酸二酯酶的第一个报道的变构抑制剂。最近，我们还完成了用手稿进行制备的磷酸二酯酶TDP2抑制剂的屏幕。此外，PCMB继续增加生物物理筛选方法的实施，以鉴定蛋白质和RNA结构的热稳定性的特定调节剂。最初，这项工作导致鉴定出一种新的小分子支架，从而改变了HIV-1 TAR RNA的稳定性。现在已经扩展了这项技术，以允许以高吞吐量格式对纯化合物和天然产物样品进行便捷筛选，以鉴定RNA和蛋白质稳定性的调节剂。最近发表的一个例子是鉴定自然产物，该天然产物有选择地调节了MIR-21的热稳定性。我们最近完成了两个针对嵌合激酶DNAJB1-PRKACA（PKADJ）和肌醇1,3,4-三磷酸5/6激酶（ITPK1）的新酶筛选，以及四个生物物理筛选；针对免疫学上的代谢酶免疫反应基因1（IRG1），Glypican-3（GPC-3）（GPC-3）的诊断靶标和SARS-COV-2受体结合结构域（RBD）和DNA读取蛋白BRD4。 PCMB还评估了从MTP筛选中针对大分子靶标的特定相互作用的化合物。我们已经使用生物物理和生化技术来评估MIR-21 RNA，TDP1，MALT1，CBLB和SARS-COV-2病毒进入的抑制剂。

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Antiviral lectins: Selective inhibitors of viral entry.

DOI：
10.1016/j.antiviral.2017.03.007
发表时间：
2017-06
期刊：
Antiviral research
影响因子：
7.6
作者：
Mitchell CA;Ramessar K;O'Keefe BR
通讯作者：
O'Keefe BR

Dentithecamides A-H, Diacylated Zoanthoxanthin Derivatives with PAX3-FOXO1 Inhibitory Activity from the Hydroid Dentitheca habereri.

Dentithecamides A-H，来自水螅 Dentitheca habereri 的具有 PAX3-FOXO1 抑制活性的二酰化 Zoanthoxanthin 衍生物。

DOI：
10.1021/acs.jnatprod.2c00246
发表时间：
2022
期刊：
Journal of natural products
影响因子：
5.1
作者：
Jiang,Wei;Tian,Xiangrong;Wang,Dongdong;Bokesch,HeidiR;Thomas,CherylL;Woldemichael,GirmaM;Gryder,BerkleyE;Wei,JunS;Song,YoungK;Chou,Hsien-Chao;Khan,Javed;O'Keefe,BarryR;Gustafson,KirkR
通讯作者：
Gustafson,KirkR

Suberitamides A-C, Aryl Alkaloids from a Pseudosuberites sp. Marine Sponge that Inhibit Cbl-b Ubiquitin Ligase Activity.

DOI：
10.3390/md18110536
发表时间：
2020-10-28
期刊：
Marine drugs
影响因子：
5.4
作者：
Kim CK;Wang D;Wilson BAP;Saurí J;Voeller D;Lipkowitz S;O'Keefe BR;Gustafson KR
通讯作者：
Gustafson KR

Sinularamides A-G, Terpenoid-Derived Spermidine and Spermine Conjugates with Casitas B-Lineage Lymphoma Proto-Oncogene B (Cbl-b) Inhibitory Activities from a Sinularia sp. Soft Coral.

DOI：
10.1021/acs.jnatprod.1c00367
发表时间：
2021-06-25
期刊：
JOURNAL OF NATURAL PRODUCTS
影响因子：
5.1
作者：
Jiang, Wei;Wang, Dongdong;Wilson, Brice A. P.;Voeller, Donna;Bokesch, Heidi R.;Smith, Emily A.;Lipkowitz, Stanley;O'Keefe, Barry R.;Gustafson, Kirk R.
通讯作者：
Gustafson, Kirk R.

Rare Caulamidine Hexacyclic Alkaloids from the Marine Ascidian Polyandrocarpa sp.