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: 8049747
• 负责人: Patrick M Woster
• 金额: $ 25.9万
• 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/8049747
• 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; Principal Investigator; Probability; Proteins; Publishing; Recombinants; Regulation; Reporting; Role; Route; Screening procedure; Sequence Homology; Series; Specificity; Structure; Tranylcypromine; Trichostatin A; Tumor Cell Line; Tumor Suppressor Genes; Tumor Suppressor Proteins; Xenograft Model; 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; programs; promoter; public health relevance; research study; small molecule libraries; tumor growth; tumor initiation; tumor xenograft; tumorigenesis; virtual

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The recently discovered enzyme lysine-specific demethylase 1 (LSD1) has been shown to play an important role in epigenetic control of gene expression, and elevated levels of LSD1 lead to a reduced expression of tumor suppressor factors that are important in human cancer. Our group has discovered a series of guanidines and biguanides that act as potent LSD1 inhibitors and reactivate these tumor suppressor genes, making these compounds a potential new class of antitumor agents. In this proposal, we describe the design and chemical synthesis of multiple new series of related analogues that will reversibly inhibit or irreversibly inactivate LSD1, and propose experiments to determine their effects on gene expression, cellular function and their ability to inhibit tumor cell growth in culture and in a mouse xenograft model.

描述（由申请人提供）：最近发现的赖氨酸特异性脱甲基酶1（LSD 1）阐明了基因表达表观遗传控制的重要细胞机制。特别地，二甲基赖氨酸4，组蛋白H3（H3 K4 me 2）是基因启动子处的转录激活染色质标记，并且LSD 1引起的该标记的异常去甲基化可能广泛地抑制在人类癌症中重要的肿瘤抑制基因的表达。我们和其他人已经进行了研究，证实LSD 1是一个令人兴奋的新治疗靶点。我们最近报道了一系列的（双）胍和（双）双胍，是重组人LSD 1的有效抑制剂。这些抑制剂显著增加H3 K4 me 2水平，启动染色质重塑并诱导肿瘤抑制基因的再表达，使其成为类似物开发的合适先导。我们是第一个在体外证明LSD 1抑制剂的抗肿瘤作用，最近已经证明了它们在体内的显着抗肿瘤作用。这些研究提供了证据或原理，即抑制LSD 1可导致显著的抗肿瘤作用。该提案的中心假设是，可以鉴定和开发抑制LSD 1的化合物用于治疗人类癌症。沿着这些思路，本提案的具体目标是：作为LSD 1潜在抑制剂的多系列类似物的设计和合成。我们将使用系统的药物化学方法，包括类似物合成和高通量评价，以产生合理设计的小分子LSD 1抑制剂库。这些类似物将在结构上与我们的先导化合物或LSD 1底物相关。我们还将使用虚拟筛选来识别商业数据库中的线索，并建议更有效的类似物进行合成和筛选。具体目标2。新合成的类似物作为LSD 1抑制剂的评价及其在培养的肿瘤细胞中的表观遗传效应的研究。将每种类似物作为纯化的LSD 1的抑制剂进行评价，并测定抑制动力学。将在HCT 116肿瘤细胞系中监测所有类似物的细胞效应。将单独评价每种化合物，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联合评价。我们将监测特定的染色质标记和基因产物，以确定每种化合物是否会引起肿瘤抑制基因的重新表达，并测量细胞生长和活力。具体目标3。LSD 1抑制剂和组合治疗的体内评价。有前途的化合物和联合治疗将推进到人类HCT 116肿瘤异种移植物的剂量和疗效研究。使用这种方法，有很高的可能性，确定有效的LSD 1抑制剂，有可能成为一个重要的新一类抗肿瘤药物。 公共卫生关系：最近发现的酶赖氨酸特异性脱甲基酶1（LSD 1）已被证明在基因表达的表观遗传控制中发挥重要作用，LSD 1水平升高导致在人类癌症中重要的肿瘤抑制因子表达减少。我们的团队已经发现了一系列胍和双胍，它们可以作为有效的LSD 1抑制剂并重新激活这些肿瘤抑制基因，使这些化合物成为潜在的新型抗肿瘤药物。在这个提议中，我们描述了多个新系列的相关类似物的设计和化学合成，这些类似物将可逆地抑制或不可逆地抑制LSD 1，并提出实验来确定它们对基因表达，细胞功能及其在培养物和小鼠异种移植模型中抑制肿瘤细胞生长的能力的影响。