Structure-based design of novel low molecular weight c-Myc inhibitors

基于结构的新型低分子量 c-Myc 抑制剂的设计

• 批准号: 8469738
• 负责人: Edward Victor Prochownik
• 金额: $ 49.37万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8469738
• 关键词: Binding; Binding Sites; Biochemical; Biological; Biological Assay; Cell Proliferation; Cells; Computer Simulation; Computing Methodologies; Dimerization; Disease; Drug Kinetics; Drug or chemical Tissue Distribution; Generations; Health; Heterodimerization; Human; In Vitro; Institution; Link; Malignant Neoplasms; Mediating; Metabolism; Modeling; Modification; Molecular; Molecular Abnormality; Molecular Conformation; Molecular Weight; NMR Spectroscopy; Neoplasms; Normal Cell; Oncogene Proteins; Oncogenes; Outcome Study; Pediatric Oncologist; Regulation; Research Personnel; Series; Site; Specificity; Structural Models; Structure; Testing; Toxic effect; Xenograft Model; ZIP protein; analog; base; c-myc Genes; design; efficacy testing; improved; in vivo; inhibitor/antagonist; insight; member; monomer; multidisciplinary; novel; pre-clinical; prevent; small molecule; synthetic peptide; therapeutic target; three dimensional structure; three-dimensional modeling; transcription factor; tumor

DESCRIPTION (provided by applicant): De-regulation of the C-MYC oncogene is among the most frequent molecular abnormalities in human cancer. c-Myc is a bHLH-ZIP transcription factor whose dimerization with another bHLH-ZIP protein, Max, is necessary for transformation and all other biological activities. That c-Myc is indispensable for cell proliferation, that most normal cells are non-cycling and thus non-c-Myc-expressing, and that transient inhibition of c-Myc may be sufficient to achieve permanent tumor regression, makes therapeutic targeting of the oncoprotein highly appealing. We have previously identified 7 low molecular weight compounds that abrogate/prevent bHLH-ZIP-mediated c-Myc-Max heterodimerization ("Myc compounds"). Non structure- guided (i.e. "unbiased") modifications of one of these has resulted in the generation of "2nd generation" analogs with up to 8-fold enhanced potencies, thus demonstrating that significant improvements in compound efficacy can be obtained even in the absence of a model of the compound in association with its target site. In order to identify even better analogs in a more rational and meaningful way, we have demonstrated that 3 of our original Myc compounds bind to distinct sites on the intrinsically disordered c- Myc bHLH-ZIP monomer, alter its secondary structure, and prevent its association with Max. The remaining 4 compounds are redundant and bind to one of these sites. Using NMR spectroscopy, we have generated structural models of the first three compounds in association with synthetic peptides comprising their cognate binding sites. Thus, in Specific Aim 1, we will utilize computational methods and high throughput in silico screens to identify new and more potent "3rd generation" analogs whose structures are rationally predicated on those of the original 3 founding members. In Specific Aim 2, we will compare the abilities of these new compounds to inhibit c-Myc-Max association and c-Myc-mediated neoplastic growth. In Specific Aim 3, we will conduct in vivo pharmacologic testing of the most promising analogs prioritized by their "ADMET" profiles. Finally, in Specific Aim 4, selected analogs identified in Specific Aims 2 and 3 will be chemically linked in order to generate "4th generation" compounds capable of simultaneous binding to two distinct sites on the c-Myc bHLH-ZIP domain. This proposal is highly interdisciplinary and interactive in that it utilizes the expertise of a pediatric oncologist/molecular biologist (Prochownik), an organic chemist (Metallo), computational biologists (Behar and Mustata), and pharmacologists (Lazo, Eiseman and Egorin). Together, this diverse, complementary, and synergistic group of investigators, from two institutions, proposes studies that promise new insights regarding the rational design of Myc compounds and the mechanisms by which they disable this critical oncoprotein.

描述（由申请方提供）：C-MYC癌基因的失调是人类癌症中最常见的分子异常之一。c-Myc是一种bHLH-ZIP转录因子，其与另一种bHLH-ZIP蛋白Max的二聚化是转化和所有其他生物活性所必需的。c-Myc对于细胞增殖是不可或缺的，大多数正常细胞是非周期性的，因此不表达c-Myc，并且c-Myc的短暂抑制可能足以实现永久性肿瘤消退，这使得针对癌蛋白的治疗靶向非常有吸引力。我们先前已经鉴定了7种消除/防止bHLH-ZIP介导的c-Myc-Max异源二聚化的低分子量化合物（“Myc化合物”）。这些之一的非结构引导的（即“无偏的”）修饰已经导致产生具有高达8倍增强的效力的“第二代”类似物，因此证明即使在不存在与其靶位点相关的化合物的模型的情况下也可以获得化合物功效的显著改善。为了以更合理和有意义的方式鉴定甚至更好的类似物，我们已经证明了我们的3种原始Myc化合物结合到固有无序的c-Myc bHLH-ZIP单体上的不同位点，改变其二级结构，并防止其与Max缔合。其余4种化合物是多余的，与这些位点之一结合。使用NMR光谱，我们已经生成了前三个化合物的结构模型与合成肽，包括其同源结合位点。因此，在具体目标1中，我们将利用计算方法和高通量计算机筛选来鉴定新的和更有效的“第三代”类似物，其结构合理地预测了原始3个创始成员的结构。在具体目标2中，我们将比较这些新化合物抑制c-Myc-Max结合和c-Myc介导的肿瘤生长的能力。在具体目标3中，我们将对最有前途的类似物进行体内药理学测试，这些类似物按其“ADMET”特征进行优先排序。最后，在特定目的4中，特定目的2和3中鉴定的选定类似物将被化学连接，以产生能够同时结合c-Myc bHLH-ZIP结构域上的两个不同位点的“第4代”化合物。该提案是高度跨学科和互动的，因为它利用了儿科肿瘤学家/分子生物学家（Prochownik），有机化学家（Metallo），计算生物学家（Behar和Mustata）和药理学家（Lazo，Escheman和Egorin）的专业知识。总之，这个来自两个机构的多样化，互补和协同的研究小组提出了一些研究，这些研究有望对Myc化合物的合理设计以及它们使这种关键癌蛋白失活的机制产生新的见解。

项目成果

Pharmacophore identification of c-Myc inhibitor 10074-G5.

• DOI: 10.1016/j.bmcl.2012.10.013
• 发表时间: 2013-01-01
• 期刊: BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
• 影响因子: 2.7
• 作者: Yap, Jeremy L.;Wang, Huabo;Hu, Angela;Chauhan, Jay;Jung, Kwan-Young;Gharavi, Robert B.;Prochownik, Edward V.;Fletcher, Steven
• 通讯作者: Fletcher, Steven

A quantitative, surface plasmon resonance-based approach to evaluating DNA binding by the c-Myc oncoprotein and its disruption by small molecule inhibitors.

一种基于表面等离子共振的定量方法，用于评估 c-Myc 癌蛋白对 DNA 的结合以及小分子抑制剂对 DNA 的破坏。

• DOI: 10.14440/jbm.2015.54
• 发表时间: 2015
• 期刊: Journal of biological methods
• 作者: Wang,Huabo;Ramakrishnan,Anand;Fletcher,Steven;Prochownik,EdwardV
• 通讯作者: Prochownik,EdwardV

MicroRNA-based screens for synthetic lethal interactions with c-Myc.

• DOI: 10.14800/rd.1330
• 发表时间: 2016-05
• 期刊: RNA & disease
• 作者: Youjun Li;Yahui Zhu;E. Prochownik

Discovery of methyl 4'-methyl-5-(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)-[1,1'-biphenyl]-3-carboxylate, an improved small-molecule inhibitor of c-Myc-max dimerization.

• DOI: 10.1002/cmdc.201402189
• 发表时间: 2014-10
• 期刊: CHEMMEDCHEM
• 影响因子: 3.4
• 作者: Chauhan, Jay;Wang, Huabo;Yap, Jeremy L.;Sabato, Philip E.;Hu, Angela;Prochownik, Edward V.;Fletcher, Steven
• 通讯作者: Fletcher, Steven

Disruption of Myc-Max heterodimerization with improved cell-penetrating analogs of the small molecule 10074-G5.

• DOI: 10.18632/oncotarget.1108
• 发表时间: 2013-06
• 期刊: Oncotarget
• 作者: Wang H;Chauhan J;Hu A;Pendleton K;Yap JL;Sabato PE;Jones JW;Perri M;Yu J;Cione E;Kane MA;Fletcher S;Prochownik EV
• 通讯作者: Prochownik EV