Identification of LSD1 inhibitors targeting epigenetic regulation in tumor cells

针对肿瘤细胞表观遗传调控的 LSD1 抑制剂的鉴定

• 批准号: 8444579
• 负责人: Patrick M Woster
• 金额: $ 23.26万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8444579
• 关键词: Active Sites; Affinity; Amino Acids; Animals; Azacitidine; Biguanides; Binding; Biological; Biological Assay; Carcinoma; Catalytic Domain; Cell Survival; Cell physiology; Chemicals; Chromatin; Colon Carcinoma; Colonic Adenoma; Computer Simulation; Cultured Tumor Cells; DNA Methyltransferase Inhibitor; Data; Databases; Development; Dose; Enzymes; Epigenetic Process; Evaluation; Family; GATA4 transcription factor; Gene Expression; Gene Expression Regulation; Gene Silencing; Genes; Genetic Transcription; Goals; Guanidines; HCT116 Cells; Histone Deacetylase Inhibitor; Histone H3; Homologous Gene; Human; In Situ; In Vitro; Inhibitory Concentration 50; Kinetics; Lead; Lethal Dose 50; Libraries; Lysine; MS-275; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Methyltransferase; Molecular Weight; Monitor; Monoamine Oxidase A; Monoamine Oxidase B; Monoamine Oxidase Inhibitors; Mus; Nitrogen; Nude Mice; Outcome; Oxidases; Peptides; Pharmaceutical Chemistry; Play; Probability; Proteins; Publishing; Recombinants; Regulation; Reporting; Role; Route; Sequence Homology; Series; Specificity; Structure; Tranylcypromine; Trichostatin A; Tumor Cell Line; Tumor Suppressor Genes; Tumor Suppressor Proteins; Xenograft Model; abstracting; analog; antitumor agent; base; cell growth; chemical synthesis; chemotherapeutic agent; chromatin modification; chromatin remodeling; cyclopropylamine; demethylation; design; high throughput screening; human lysine specific demethylase 1; in vivo; inhibitor/antagonist; lysine analog; neoplastic cell; new therapeutic target; peptidomimetics; polyamine oxidase; preclinical study; promoter; research study; screening; small molecule libraries; tumor growth; tumor initiation; tumor xenograft; tumorigenesis; virtual

Abstract. The recent discovery of the enzyme lysine-specific demethylase 1 (LSD1) has illuminated an important cellular mechanism for epigenetic control of gene expression. In particular, dimethyl lysine 4, histone H3 (H3K4me2) is a transcription activating chromatin mark at gene promoters, and aberrant demethylation of this mark by LSD1 may broadly repress the expression of tumor suppressor genes that are important in human cancer. We and others have conducted studies verifying that LSD1 is an exciting new therapeutic target. We recently reported a series of (bis)guanidines and (bis)biguanides that are potent inhibitors of recombinant human LSD1. These inhibitors significantly increase H3K4me2 levels, initiate chromatin remodeling and induce the re-expression of tumor suppressor genes, making them suitable leads for analogue development. We were the first to demonstrate antitumor effects of LSD1 inhibitors in vitro and have recently demonstrated their significant antitumor effects in vivo. These studies provide proof or principle that inhibition of LSD1 can lead to significant antitumor effects. The central hypothesis of this proposal is that compounds that inhibit LSD1 can be identified and developed for use in the treatment of human cancer. Along these lines, the specific aims of this proposal are: Specific aim 1. Design and synthesis of multiple series of analogues as potential inhibitors of LSD1. We will use a systematic medicinal chemistry approach that includes analogue synthesis and high-throughput evaluation to produce rationally designed libraries of small-molecule LSD1 inhibitors. These analogues will be structurally related to our lead compounds, or to the LSD1 substrate. We will also use virtual screening to identify leads from commercial databases, and to suggest more potent analogues for synthesis and screening. Specific aim 2. Evaluation of newly synthesized analogues as LSD1 inhibitors and study of their epigenetic effects in cultured tumor cells. Each analogue will be evaluated as an inhibitor of purified LSD1, and the kinetics of inhibition will be determined. The cellular effects of all analogues will be monitored in the HCT116 tumor cell line. Each compound will be evaluated alone, and in combination with a DNA methyltransferase inhibitor and/or a class I/II histone deacetylase inhibitor. We will monitor specific chromatin marks and gene products to determine whether each compound causes tumor suppressor gene re-expression, and cell growth and viability will be measured. Specific aim 3. Evaluation of LSD1 inhibitors and combination treatments in vivo. Promising compounds and combination treatments will be advanced to dosing and efficacy studies in human HCT116 tumor xenografts. Using this approach, there is a high probability of identifying potent LSD1 inhibitors that have the potential to become an important new class of antitumor agent.

抽象的。最近发现的赖氨酸特异性脱甲基酶1(LSD1)揭示了一种 表观遗传调控基因表达的重要细胞机制。尤其是二甲基赖氨酸4， 组蛋白H3(H3K4me2)是一种位于基因启动子上的转录激活染色质标记 LSD1使该标记去甲基化可能广泛抑制肿瘤抑制基因的表达 它们在人类癌症中很重要。我们和其他人已经进行了研究，证实LSD1是一种 令人振奋的新治疗目标。我们最近报道了一系列(双)胍类和(双)双胍类化合物。 它们是重组人LSD1的有效抑制剂。这些抑制剂显著增加了H3K4me2 水平，启动染色质重塑，并诱导肿瘤抑制基因的重新表达，使 它们适合用于模拟开发。我们是第一个展示其抗肿瘤作用的人 LSD1抑制剂在体外，最近已经在体内证明了它们显著的抗肿瘤作用。这些 研究提供了证据或原理，即抑制LSD1可以导致显著的抗肿瘤作用。这个 这一提议的中心假设是，可以识别出抑制LSD1的化合物并 开发用于治疗人类癌症的药物。按照这些思路，这一行动的具体目标是 建议有：具体目标1.作为潜力的多系列类似物的设计和合成 LSD1的抑制剂。我们将使用一种系统的药物化学方法，包括类似物 合理设计小分子文库的合成与高通量评价 LSD1抑制剂。这些类似物将在结构上与我们的先导化合物或LSD1相关 底物。我们还将使用虚拟筛选来从商业数据库中识别线索，并 建议更有效的类似物用于合成和筛选。具体目标2.评价新的 作为LSD1抑制剂的合成类似物及其在培养的肿瘤细胞中的表观遗传效应研究。 每个类似物都将被评估为纯化的LSD1的抑制剂，其抑制动力学将为 下定决心。所有类似物的细胞效应将在HCT116肿瘤细胞系中进行监测。每个人 化合物将单独进行评估，并与DNA甲基转移酶抑制剂和/或 I/II类组蛋白脱乙酰酶抑制剂。我们将监测特定的染色质标记和基因产品，以 确定每种化合物是否导致肿瘤抑制基因的重新表达，以及细胞生长和 生存能力将被衡量。具体目标3.对LSD1抑制剂及其联合治疗的评价 活着。有希望的化合物和联合治疗将在剂量和疗效方面取得进展 人HCT116肿瘤移植瘤的研究使用这种方法，很有可能 鉴定有可能成为一类重要的新抗肿瘤药物的有效LSD1抑制剂 探员。