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: 9378823
• 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 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9378823
• 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; Recruitment Activity; 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; mutant; neoplastic cell; next generation; novel; novel therapeutics; pressure; protein protein interaction; 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结构域(PBD)是PLK1有丝分裂功能的关键，也是底物所必需的 招募和亚细胞定位。麦金尼斯实验室已经取得了重大进展， 基于PBD抑制作用的PLK1选择性抑制剂的研究进展 抑制并保持对临床使用的三磷酸腺苷耐药的PLK1位点突变体的活性 竞争性抑制药。这些化合物将成为高价值的化学生物学探针，为进一步建立 通过PBD以非ATP竞争性方式抑制PLK1的表型后果，在验证中 PBD作为抗有丝分裂药物的靶点及其作为下一代治疗药物的潜力。这项建议是 在以下方面具有重大意义和创新性： 1)开发了一种称为替换的创新策略来生成细胞周期特异性CDK 作为抗肿瘤治疗药物的抑制剂。此外，McInnes实验室验证了这一战略 PLK1 PBD揭示被发现片段连接的肽抑制物被取代 值得尊敬的抗增殖活性和表型与靶点抑制一致。 2)针对PBD的化学生物探针对PLK1具有高度的选择性，并具有 对PLK3肿瘤抑制因子的最小亲和力。这对于进一步建立表型很重要。 选择性阻断PLK1-PBD的后果以及避免PLK3的有害影响 抑制抗肿瘤治疗药物的发展。我们已经证明了我们最大的 有效的PLK1PBD结构域抑制剂对PLK3的活性最低。 3)PBD靶向化合物将保持对已获得的肿瘤细胞的抗肿瘤活性 对基于三磷酸腺苷的抑制剂的抗药性。如上所述，单点突变(C67V)能够 使PLK1对结构不同的基于ATP的抑制剂具有抵抗力。靶向非催化功能 PLK1的使用应该会降低耐药的选择性压力，多溴联苯D抑制剂可用于 联合应用以三磷酸腺苷为基础的抑制剂作为PLK1靶向临床的协同手段。 4)目前的铅化合物在处理过的细胞中诱导了一种重塑PLK1的表型 击倒对手。这与报道的弱结合小分子PBD抑制剂形成了鲜明的对比 诱导的部分效应，可能不具有理想的抗肿瘤活性水平。我们观察到的一些人 表型是新奇的，提示后期灾难，已知的促进肿瘤选择性。 5)新型PBD抑制剂将成为肿瘤选择性化合物。最近的发现，PLK1抑制是 在PTEN缺陷、突变的RAS和p53缺陷的癌症中合成致命的将被利用来产生 肿瘤选择性化合物。非肽类抑制剂将用于细胞研究以确认肿瘤细胞 特异性、PLK1选择性和细胞表型与On靶标抑制一致。