Identification of LSD1 inhibitors targeting epigenetic regulation in tumor cells

抽象的。最近发现的赖氨酸特异性脱甲基酶1（LSD 1）阐明了一种新的脱甲基酶。 表观遗传控制基因表达的重要细胞机制。特别是二甲基赖氨酸4， 组蛋白H3（H3 K4 me 2）是基因启动子上转录激活染色质标记， LSD 1对该标记的去甲基化可能广泛抑制肿瘤抑制基因的表达， 在人类癌症中很重要。我们和其他人已经进行了研究，证实LSD 1是一种 新的治疗靶点。我们最近报道了一系列（双）胍和（双）双胍类化合物 它们是重组人LSD 1的有效抑制剂。这些抑制剂显著增加H3 K4 me 2 水平，启动染色质重塑和诱导肿瘤抑制基因的重新表达， 它们是模拟开发的合适线索。我们是第一个证明 LSD 1抑制剂在体外和最近已证明其显着的体内抗肿瘤作用。这些 研究提供了抑制LSD 1可导致显著的抗肿瘤作用的证据或原理。的 该建议的中心假设是可以鉴定出抑制LSD 1的化合物， 开发用于治疗人类癌症。沿着这些路线， 具体目标1。多系列潜在类似物的设计与合成 LSD 1的抑制剂。我们将使用系统的药物化学方法，包括类似物 以产生合理设计小分子文库 LSD 1抑制剂。这些类似物将在结构上与我们的先导化合物或LSD 1相关。 衬底我们还将使用虚拟筛选从商业数据库中识别线索， 提出了用于合成和筛选的更有效的类似物。具体目标2。评价新 作为LSD 1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，并将抑制动力学进行比较。 测定将在HCT 116肿瘤细胞系中监测所有类似物的细胞效应。每个 化合物将单独评估，以及与DNA甲基转移酶抑制剂和/或DNA甲基转移酶抑制剂组合评估。 I/II类组蛋白去乙酰化酶抑制剂。我们将监测特定的染色质标记和基因产物， 确定每种化合物是否引起肿瘤抑制基因再表达和细胞生长， 将测量可行性。具体目标3。LSD 1抑制剂和联合治疗在糖尿病中的评价 vivo.有前景的化合物和联合治疗将推进剂量和疗效 在人HCT 116肿瘤异种移植物中的研究。使用这种方法， 鉴定有潜力成为重要的新一类抗肿瘤药物的有效LSD 1抑制剂 剂

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.