Novel Chemical Biology Probes based on Selective Inhibitors of the Polo-Box Domain of PLK1

基于 PLK1 Polo-Box 结构域选择性抑制剂的新型化学生物学探针

• 批准号: 9517781
• 负责人: Campbell McInnes
• 金额: $ 7.16万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9517781
• 关键词: Active Sites; Affinity; Aftercare; Anaphase; Antimitotic Agents; Antineoplastic Agents; Binding; Biology; Catalytic Domain; Cell Cycle; Cell Line; Cell model; Cells; Chemicals; Clinic; Clinical; Clinical Trials; Data; Development; Drug Targeting; Family; Family member; Foundations; Generations; Goals; Investigation; Laboratories; Lead; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Measures; Methods; Mitosis; Mitotic; Modeling; Molecular; PLK1 gene; PLK3 gene; PTEN gene; Peptides; Permeability; Pharmaceutical Preparations; Phenotype; Phosphopeptides; Phosphorylation; Phosphotransferases; Polo-Box Domain; Prodrugs; Reporting; Resistance; Resistance development; Role; Site; Specificity; Structure-Activity Relationship; Substrate Interaction; Suggestion; TP53 gene; Therapeutic; Tumor Suppressor Proteins; Validation; antitumor agent; antitumor drug; antitumor effect; base; clinically relevant; drug discovery; high throughput screening; inhibitor/antagonist; innovation; knock-down; lead optimization; mutant; neoplastic cell; next generation; novel; novel therapeutics; pressure; protein protein interaction; recruit; small molecule; success; tumor

The Polo-box domain (PBD) is critical for mitotic functions of PLK1 and has shown to be required for substrate recruitment and sub-cellular localization. The McInnes laboratory has made significant progress toward the development of PLK1 selective inhibitors based on PBD inhibition that have minimal activity on the PLK3 tumor suppressor and which retain activity against PLK1 site mutants that are resistant to clinically utilized ATP competitive inhibitors. These compounds will be high value chemical biology probes for further establishing the phenotypic consequences of inhibiting PLK1 in a non-ATP competitive fashion through the PBD, in validating the PBD as an anti-mitotic drug target and in their potential as next generation therapeutics. This proposal is significant and innovative with respect to the following: 1) An innovative strategy called REPLACE has been developed to generate cell cycle specific CDK inhibitors as anti-tumor therapeutics. Furthermore the McInnes laboratory validated this strategy for the PLK1 PBD revealing that fragment ligated peptidic inhibitors discovered though REPLACE have respectable anti-proliferative activity and phenotypes consistent with target inhibition. 2) Chemical biology probes that target the PBD will be highly selective for PLK1 and possess minimal affinity for the PLK3 tumor suppressor. This is important to further establish the phenotypic consequences of blocking the PLK1 PBD selectively and also to avoid deleterious effects of PLK3 inhibition in the development of anti-tumor therapeutics. We have already demonstrated that our most potent PLK1 PBD domain inhibitors have minimal activity against PLK3. 3) PBD targeted compounds will retain antitumor activity against tumor cells that have acquired resistance to ATP-based inhibitors. As described above, a single point mutant (C67V) is capable of rendering PLK1 resistant to structurally diverse ATP-based inhibitors. Targeting non-catalytic functions of PLK1 should result in less selective pressure for resistance and PBD inhibitors might be used in combination with ATP-based inhibitors as a synergistic means of PLK1 targeting in the clinic. 4) Current lead compounds induce a phenotype in treated cells that recapitulates a PLK1 knockdown. This stands in contrast to weakly binding small molecule PBD inhibitors reported that only induced partial effects, and may not possess a desirable level of anti-tumor activity. Some of our observed phenotypes are novel and suggestive of anaphase catastrophe, known to promote tumor selectivity. 5) Novel PBD inhibitors will be tumor selective compounds. Recent discoveries that PLK1 inhibition is synthetically lethal in PTEN deficient, mutant Ras and p53 deficient cancers will be exploited to generate tumor selective compounds. Non-peptidic inhibitors will be used in cellular studies to confirm tumor cell specificity, PLK1 selectivity, and cellular phenotypes consistent with on target inhibition.

polo-box域（PBD）对于PLK1的有丝分裂功能至关重要，并且已证明是底物所需的 招聘和细胞局部定位。麦金尼斯实验室已取得了重大进展 基于对PLK3肿瘤活性最小的PBD抑制剂的PLK1选择性抑制剂的开发 抑制剂并保留对PLK1位点突变体的活性 竞争性抑制剂。这些化合物将是高价值的化学生物学探针，以进一步确定 通过PBD以非ATP竞争方式抑制PLK1的表型后果，验证 PBD作为抗溶毒靶靶标的及其潜力作为下一代治疗剂。该提议是 关于以下内容的重要性和创新性： 1）已经开发了一种称为替代的创新策略来产生细胞周期特定的CDK 抑制剂作为抗肿瘤疗法。此外，麦金尼斯实验室还确认了这一策略 PLK1 PBD揭示了碎片结扎的肽抑制剂，但替换已发现 与目标抑制一致的可观的抗增殖活性和表型。 2）靶向PBD的化学生物学探针对PLK1具有很高的选择性并具有 PLK3肿瘤抑制剂的最小亲和力。这对于进一步建立表型很重要 有选择地阻止PLK1 PBD的后果，也避免PLK3的有害影响 抑制抗肿瘤疗法的发展。我们已经证明了我们的最大 有效的PLK1 PBD结构域抑制剂对PLK3的活性很小。 3）PBD靶向化合物将保留对已获得的肿瘤细胞的抗肿瘤活性 对基于ATP的抑制剂的抗性。如上所述，单点突变体（C67V）能够 对基于ATP的抑制剂具有抗性PLK1。靶向非催化功能 PLK1的抗药性应降低，并且可以使用PBD抑制剂的选择性较小 结合基于ATP的抑制剂作为诊所中PLK1靶向的协同手段。 4）当前铅化合物诱导经过概括PLK1的处理细胞中的表型 击倒。这与弱结合的小分子PBD抑制剂相反，只有 诱导的部分作用，可能没有理想水平的抗肿瘤活性。我们观察到的一些 表型是新颖的，暗示了后期灾难，已知可以促进肿瘤选择性。 5）新型的PBD抑制剂将是肿瘤选择性化合物。最近发现PLK1抑制是 将利用缺乏症状，突变体Ras和p53缺乏癌症的综合致命。 肿瘤选择性化合物。非肽抑制剂将用于细胞研究以证实肿瘤细胞 特异性，PLK1选择性和细胞表型与目标抑制一致。