Structure-based In silico Screen for Small Molecule Inhibitors of Ets-1 Activity

描述（由申请人提供）：炎症过程有助于导致自身免疫性疾病（包括类风湿关节炎（RA））组织破坏的病理事件。对于患有RA的患者来说，能够阻断TNF1和IL-1的治疗剂的发展是重要的治疗里程碑，然而，大量患者未能对这些治疗作出反应，可能是由于它们固有的无法抑制这种复杂疾病所需的其他途径。一些实验室认为，为了进一步了解RA的病理生理，需要确定、靶向和验证新的治疗靶点。一类潜在的靶标是细胞因子诱导的转录因子；NF-k B、AP-1、C/EBP、ETS-1均在RA滑膜中检测到。最近的一些出版物支持靶向转录因子（TFs）的重要性，这将提供一种通过受控的变阻器“开关”而不是二元开/关机制重新调节基因表达途径的机制，包括细胞因子途径。此外，丰富的信息和对转录生物学和基因调控的更好理解支持tf本身是治疗干预的潜在靶点。拟议的项目是高度创新的，将我们基于转化发现的科学与体外和体内验证这一新的靶标空间结合起来，TF-DNA界面将首次用于开发男性类风湿性关节炎治疗。这一协作和真正的协同应用涉及具有互补技能的研究人员，利用基于硅结构的小分子发现、基于核磁共振波谱的靶标验证和体外表征/评估的优势，支持我们识别和开发新的小分子的方法，这些小分子专门靶向和抑制Ets-1与其序列特异性DNA启动子元件之间的相互作用界面。这些研究人员具有独特的优势和专业知识，当他们合作时，就会为创造性的、“开箱即用”的思想和执行提供重要的机会，正如支持该应用程序的重要初步数据所证明的那样。具有可证明活性的小分子，如已确定的那些，为TF依赖性转录调控提供了一个有吸引力的机会，为在TF- dna水平上选择性抑制ETS TF活性的治疗剂的“突破先导”开发提供了一种创新策略。该应用侧重于选择性靶向这种TF-DNA界面，这代表了一种新的tf -1依赖性转录调控方法，并为开发选择性靶向这种转录因子的治疗剂提供了独特的机会。通过对核因子κ B （NF-kB）的特异性转录调控，小分子抑制TF-DNA相互作用界面为RA的转录治疗提供了一个有希望的范式转变。通过丰富的信息和对转录生物学和基因调控的更好理解，包括NF-kB、HIF112等在内的tf已成为治疗干预的新靶点。拟议的项目是高度创新的，将我们基于转化发现的科学与新目标空间的体外和体内验证相结合；TF-DNA界面用于RA新疗法的鉴定和后续开发。tf是基因表达的调节因子，因此是多种生物过程所必需的，包括生长、分化和发育，以及癌症和/或炎症等病理过程。序列特异性TF-DNA相互作用在空间和时间上受到调控，从而在蛋白质- dna界面上产生精细的特异性和选择性。选择性靶向和抑制TF-DNA复合物相互作用界面的能力代表了一种新的、高度特异性的策略，用于重编程在RA和癌症中失调的特定基因通路。与其他方法不同的是；聚酰胺、人工转录因子、锌指蛋白疗法，我们建议在硅、虚拟筛选或高通量对接（HTD）方面进行合作，以筛选可公开访问的大型化学库，以选择性地靶向并抑制这种明确定义的分子界面。重要的是，硅HTD非常适合探索新的目标空间，如Ets-1 TF-DNA相互作用界面。我们的中心假设是，靶向并破坏这种界面的小分子将能够调节异常基因转录，从而为RA的治疗提供一种新的范例。这些具有显著体内活性的小分子支架将为RA的转录治疗提供一个平台。这种方法如果成功，将代表一个创新的、非常独特的机会，可以开发一种“首次在人体中”的治疗方法，通过靶向TF的选择性/特异性界面，选择性地靶向和破坏下游TF介导的基因表达。

DESCRIPTION (provided by applicant): Inflammatory processes contribute to the pathological events that lead to tissue destruction in autoimmune diseases including rheumatoid arthritis (RA). For patients suffering with RA the development of therapeutic agents that are capable of blocking TNF1 and IL-1 have been important therapeutic milestones, however a significant number of patients fail to respond to these therapies possibly due to their inherent inability to inhibit other pathways requisite in this complex disease. Several laboratories have suggested that in order to further understand the pathophysiologies of RA new therapeutic targets need to be identified, targeted and validated. One potential class of targets is the cytokine-induced transcription factors; NF-k B, AP-1, C/EBP, and ETS-1 all of which have been detected in RA synovium. Several recent publications support the importance of targeting transcription factors (TFs), which would provide a mechanism of re-regulating gene expression pathways including the cytokine pathway through a controlled, rheostat "switch" rather than a binary on/off mechanism. In addition, a wealth of information and a better understanding of transcriptional biology and gene regulation supports that TFs themselves are potential targets for therapeutic intervention. The proposed project is highly innovative, partnering our translational discovery-based science with in vitro and in vivo validation of this novel target space, the TF-DNA interface for use in the development of first in man type RA therapy. This collaborative and truly synergistic application involving investigators with complementary skills partners the strengths of in silico structure-based small molecule discovery with NMR spectroscopy based target validation and in vitro characterization/evaluation in support of our approach for identifying and developing novel small molecules that specifically target and inhibit the interaction interface between Ets-1 and its sequence specific DNA promoter element. These investigators have unique strengths and expertise, which when partnered provides a significant opportunity for creative, "out of the box" thought and execution as demonstrated by the significant preliminary data in support of this application. Small molecules with demonstrable activity such as those identified represent an attractive opportunity for TF-dependent transcriptional regulation, providing an innovative strategy for the "hit-through-lead" development of therapeutic agents that selectively inhibit ETS TF activity at the TF-DNA level. This application is focused on selectively targeting this TF-DNA interface, which represents a novel approach for TF-dependent transcriptional regulation of Ets-1 and provides a unique opportunity for the development of therapeutic agents selectively targeting this transcription factor. Small molecule inhibition of the TF-DNA interaction interface provides a promising paradigm shift in transcriptional therapy for RA through pathway specific transcriptional regulation as has been attempted for nuclear factor kappa B (NF-kB).Through a wealth of information and a better understanding of transcriptional biology and gene regulation, TFs including NF-kB, HIF112 and others have emerged as novel targets for therapeutic intervention. The proposed project is highly innovative, partnering our translational discovery-based science with in vitro and in vivo validation of novel target space; the TF-DNA interface for use in the identification and subsequent development of novel therapies for RA. TFs are established regulators of gene expression and as such are requisite for a variety of biological processes, including growth, differentiation and development as well as pathological processes such as cancer and/or inflammation. Sequence-specific TF-DNA interactions are spatially and temporally regulated, resulting in refined specificity and selectivity at the protein-DNA interface. The ability to selectively target and inhibit the interaction interface of the TF-DNA complex represents a novel, highly specific strategy for reprogramming specific gene pathways that are deregulated in RA and cancer. Unlike other approaches including; polyamides, artificial transcription factors, zinc finger protein therapeutics we have proposed to partner in silico, virtual screening or high throughput docking (HTD) to screen large publicly accessible chemical repositories for small molecules that selectively target and thus inhibit this well-defined molecular interface. Importantly, in silico HTD is ideally suited for the exploration of novel target space such as the Ets-1 TF-DNA interaction interface. Our central hypothesis is that small molecules that target and disrupt this interface would be capable of regulating aberrant gene transcription and would thus offer a novel paradigm of therapy for RA. Those small molecule scaffolds that demonstrate significant in vivo activity would provide a transcriptional therapy platform for RA. This approach, if successful, represents an innovative and very unique opportunity to develop a "first in man" therapeutic approach that will selectively target and disrupt downstream TF-mediated gene expression by targeting the selectivity/specificity interface of the TF.