Different active sites of the proteasome as drug targets in cancer

蛋白酶体的不同活性位点作为癌症药物靶点

• 批准号: 8075421
• 负责人: Alexei Kisselev
• 金额: $ 26.52万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-20 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8075421
• 关键词: Active Sites; Address; Apoptosis; Apoptotic; Bortezomib; Breast Cancer Cell; Breast Cancer Model; Cancer cell line; Caspase; Catalytic Domain; Cell Line; Cell Proliferation; Cells; Clinical Treatment; Clinical Trials; Data; Development; Disease; Drug Delivery Systems; Epithelial Cells; Estrogen receptor negative; Goals; Growth; Human; Malignant Neoplasms; Mammary gland; Molecular; Molecular Target; Multiple Myeloma; Non-Malignant; Pathway interactions; Peripheral Blood Lymphocyte; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase I Clinical Trials; Play; Property; Proteasome Inhibitor; Proteins; Public Health; Quality Control; Research Personnel; Resistance; Review Literature; Role; Site; Specificity; Staging; Structure; Testing; Therapeutic; Toxic effect; Trypsin; Ubiquitin; Work; base; cancer cell; cancer therapy; cancer type; caspase-2; chymotrypsin; cytotoxic; cytotoxicity; design; inhibitor/antagonist; killings; malignant breast neoplasm; multicatalytic endopeptidase complex; neoplastic; novel; pre-clinical; programs; protein degradation; salinosporamide A; tumorigenesis

DESCRIPTION (provided by applicant): Protein degradation by proteasomes plays an essential role in the proliferation of malignant cells. The proteasome inhibitor VELCADE (bortezomib, PS-341) is being used for the treatment of multiple myeloma and is in clinical trials for the treatment of other cancers. The proteasome has three types of active sites: chymotrypsin-like (Beta 5), trypsin-like (Beta 2), and caspase-like (Beta 1). Our long-term goal is to understand the precise roles of these active sites as targets of anti-neoplastic agents and to use this information to design new, more potent, less toxic drugs of this class. Bortezomib was developed as an inhibitor of the Beta 5 site, which has long been considered the only suitable target for inhibition. However, bortezomib also inhibits the (Beta 1 site, and we have recently obtained data showing that inhibition of the Beta 1 site is needed to achieve maximal cytotoxicity. Based on these data and on critical review of the literature, we hypothesize that (a) all three active sites are drug molecular targets, (b) inhibition of at least two sites is required to achieve optimal cytotoxicity, (c) the therapeutic windows of proteasome inhibitors depend on which active sites they target, and (d) the exact pathways by which proteasome inhibitors induce apoptosis in malignant cells depend on which active sites they target. We will test this hypothesis in multiple myeloma and breast cancer cells using unique, cell-permeable, specific inhibitors of proteasome catalytic sites that we have developed and are developing. The specific aims of this proposal are as follows: (1) To determine the growth inhibitory and cytotoxic effects of specific inhibitors of (Beta 5, Beta1, and Beta 2 catalytic sites, alone and in combination with each other, on cells derived from multiple myeloma and breast cancers, on immortalized and transformed human mammary epithelial cells, and on human peripheral blood lymphocytes; (2) To determine whether the mechanisms by which proteasome inhibitors kill multiple myeloma cells depend on the catalytic sites being targeted by these agents; The results of these studies will define what active-site pharmacological specificities proteasome inhibitors must have to achieve optimal anti-neoplastic activity and therapeutic windows. Taken together, these studies will provide a strong rationale for development of novel inhibitors with desired molecular pharmacological properties. Such compounds have potentially broad applicability for cancer therapy.

描述（由申请人提供）：蛋白酶体的蛋白质降解在恶性细胞的增殖中起着重要作用。蛋白酶体抑制剂 VELCADE（硼替佐米，PS-341）用于治疗多发性骨髓瘤，并正在进行治疗其他癌症的临床试验。蛋白酶体具有三种类型的活性位点：胰凝乳蛋白酶样 (Beta 5)、胰蛋白酶样 (Beta 2) 和半胱天冬酶样 (Beta 1)。我们的长期目标是了解这些活性位点作为抗肿瘤药物靶标的精确作用，并利用这些信息设计新的、更有效、毒性更低的此类药物。硼替佐米被开发为 Beta 5 位点的抑制剂，长期以来一直被认为是唯一合适的抑制靶点。然而，硼替佐米也会抑制 (Beta 1 位点，我们最近获得的数据表明，需要抑制 Beta 1 位点才能实现最大的细胞毒性。根据这些数据和对文献的严格审查，我们假设 (a)所有三个活性位点都是药物分子靶标，(b) 需要抑制至少两个位点才能实现最佳细胞毒性，(c) 蛋白酶体抑制剂的治疗窗口取决于它们靶向的活性位点，以及(d) 蛋白酶体抑制剂诱导恶性细胞凋亡的确切途径取决于它们靶向的活性位点，我们将使用我们研究的独特的、细胞可渗透的、特异性的蛋白酶体催化位点抑制剂在多发性骨髓瘤和乳腺癌细胞中测试这一假设。该提案的具体目标如下：(1) 确定（Beta 5、Beta1 和 Beta 2 催化位点单独和）的特定抑制剂的生长抑制和细胞毒性作用。彼此组合，作用于源自多发性骨髓瘤和乳腺癌的细胞、作用于永生化和转化的人乳腺上皮细胞、以及作用于人外周血淋巴细胞； (2) 确定蛋白酶体抑制剂杀死多发性骨髓瘤细胞的机制是否取决于这些药物靶向的催化位点；这些研究的结果将确定蛋白酶体抑制剂必须具有哪些活性位点药理学特异性才能实现最佳的抗肿瘤活性和治疗窗。总而言之，这些研究将为开发具有所需分子药理学特性的新型抑制剂提供强有力的理论依据。此类化合物对于癌症治疗具有潜在广泛的适用性。

项目成果

Selective inhibitor of proteasome's caspase-like sites sensitizes cells to specific inhibition of chymotrypsin-like sites.

• DOI: 10.1016/j.chembiol.2009.11.015
• 发表时间: 2009-12-24
• 期刊: Chemistry & biology
• 作者: Britton M;Lucas MM;Downey SL;Screen M;Pletnev AA;Verdoes M;Tokhunts RA;Amir O;Goddard AL;Pelphrey PM;Wright DL;Overkleeft HS;Kisselev AF
• 通讯作者: Kisselev AF

Azido-BODIPY acid reveals quantitative Staudinger-Bertozzi ligation in two-step activity-based proteasome profiling.

叠氮基-BODIPY 酸揭示了基于两步活性的蛋白酶体分析中的定量 Staudinger-Bertozzi 连接。

• DOI: 10.1002/cbic.200800231
• 发表时间: 2008
• 期刊: Chembiochem : a European journal of chemical biology
• 作者: Verdoes,Martijn;Florea,BogdanI;Hillaert,Ulrik;Willems,LianneI;vanderLinden,WouterA;Sae-Heng,Myra;Filippov,DmitriV;Kisselev,AlexeiF;vanderMarel,GijsbertA;Overkleeft,HermanS
• 通讯作者: Overkleeft,HermanS

Cell-line-specific high background in the Proteasome-Glo assay of proteasome trypsin-like activity.

蛋白酶体胰蛋白酶样活性的 Proteasome-Glo 测定中的细胞系特异性高背景。

• DOI: 10.1016/j.ab.2014.01.009
• 发表时间: 2014
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: Wilkins,OwenM;Downey,SondraL;Weyburne,EmilyS;Williams,DavidA;Mirabella,AnneC;Overkleeft,HermanS;Kisselev,AlexeiF
• 通讯作者: Kisselev,AlexeiF

Discovery of a potent and highly β1 specific proteasome inhibitor from a focused library of urea-containing peptide vinyl sulfones and peptide epoxyketones.

从含尿素肽乙烯基砜和肽环氧酮的重点库中发现一种有效且高度β1 特异性的蛋白酶体抑制剂。

• DOI: 10.1039/c1ob06554h
• 发表时间: 2012
• 期刊: Organic & biomolecular chemistry
• 影响因子: 3.2
• 作者: vanderLinden,WouterA;Willems,LianneI;Shabaneh,TamerB;Li,Nan;Ruben,Mark;Florea,BogdanI;vanderMarel,GijsA;Kaiser,Markus;Kisselev,AlexeiF;Overkleeft,HermanS
• 通讯作者: Overkleeft,HermanS

Incorporation of non-natural amino acids improves cell permeability and potency of specific inhibitors of proteasome trypsin-like sites.

• DOI: 10.1021/jm3016987
• 发表时间: 2013-02-14
• 期刊: JOURNAL OF MEDICINAL CHEMISTRY
• 影响因子: 7.3
• 作者: Geurink, Paul P.;van der Linden, Wouter A.;Mirabella, Anne C.;Gallastegui, Nerea;de Bruin, Gerjan;Blom, Annet E. M.;Voges, Mathias J.;Mock, Elliot D.;Florea, Bogdan I.;van der Marel, Gijs A.;Driessen, Christoph;van der Stelt, Mario;Groll, Michael;Overkleeft, Herman S.;Kisselev, Alexei F.
• 通讯作者: Kisselev, Alexei F.