Small Molecule Inhibitors of Anthrax Lethal Factor

炭疽致死因子小分子抑制剂

• 批准号: 7092076
• 负责人: Norton P Peet
• 金额: $ 71.64万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7092076
• 关键词: Bacillus anthracis; X ray crystallography; anthrax; anthrax toxin; antibacterial agents; bacterial proteins; bioterrorism /chemical warfare; cell line; chemical structure function; combinatorial chemistry; cytoprotection; drug design /synthesis /production; drug screening /evaluation; laboratory rat; liver cells; macrophage; metalloendopeptidases; microorganism disease chemotherapy; microsomes; nonhuman therapy evaluation; pharmacokinetics; protease inhibitor; small molecule; structural biology

DESCRIPTION (provided by applicant): Aerosolized spores of Bacillus anthracis represent one of the most serious bio-terrorist threats to the security of the United States. New therapeutics based upon novel chemical scaffolds are vital to the biodefense armory because they are likely to be effective against both natural and engineered resistant forms of B. anthracis. Our strategy is to build effective anthrax therapeutics not by targeting the viability of the organism but by targeting the mechanism of pathogenesis directly. Toward this aim, we have identified drug-like small molecule LF inhibitors through a combination of screening of chemical libraries and 3D sub-structure database mining. These inhibitors have been shown to specifically bind to LF with low (M potencies in enzymatic assays and to protect macrophages against challenge with toxin containing LF. The identified inhibitor classes are novel and devoid of non-drug like features such as the hydroxamic acid and peptidic backbone, which are common in many known protease inhibitors. Thus, this validated hit series forms the basis for development of efficacious, safe and orally bioavailable drugs against anthrax. To further enhance the probability of success of this project, the X-ray structure of the inhibitor bound in the active site of LF has been solved thus providing a powerful tool, which we will use to guide the inhibitor refinement process through structure-based drug design. The overall goal of this project is to develop a small molecule LF inhibitor to treat anthrax. In Phase I, we will apply proven techniques of medicinal and combinatorial chemistry; inhibitor-enzyme complex X-ray crystallography and structure-based drug design (SBDD) to rapidly synthesize and evaluate derivatives of this new validated hit series. In an iterative process, we will probe focused compound libraries for features contributing to tighter binding and more potent inhibition of LF by measuring the enzymatic, cellular activity and specificity of derivatives. We will determine the X-ray structures of improved inhibitors bound to the LF active site, and use these data to develop refined pharmacophore models to guide further probing of the structure activity relationship (SAR). In addition, we will assess compounds for optimal ADME (Absorption, Distribution, Metabolism, Elimination) properties (e.g., liver microsome stability, Cyp isoform inhibition and Caco2 permeability). Compounds with sufficient enzymatic, cellular potency and ADME properties will be synthesized at the 1-2 g level and tested for efficacy in a LF induced rat death model. Successful rescue of rats from LF-induced death will qualify compounds as in vivo-validated leads. In Phase II, we will further evaluate these leads for in vivo efficacy, pharmacokinetic properties, toxicity and safety pharmacology, in two species, in order to develop them into pre-IND clinical candidates, suitable for human clinical trials (Phase III).

描述（由申请人提供）：炭疽杆菌的气溶胶孢子是对美国安全最严重的生物恐怖威胁之一。基于新型化学支架的新疗法对于生物防御军械库至关重要，因为它们可能对天然和工程耐药形式的炭疽杆菌有效。我们的策略是建立有效的炭疽疗法，不是通过针对生物体的生存能力，而是直接针对发病机制。为了实现这一目标，我们通过化学库筛选和 3D 子结构数据库挖掘相结合，鉴定出了类药小分子 LF 抑制剂。这些抑制剂已被证明在酶测定中以低 (M) 特异性与 LF 结合，并保护巨噬细胞免受含有 LF 的毒素的攻击。所鉴定的抑制剂类别是新颖的，并且缺乏非药物样特征，例如异羟肟酸和肽因此，这一经过验证的命中系列奠定了开发有效、安全和口服生物可利用的抗炭疽药物的基础。该项目成功的可能性在于，结合在 LF 活性位点的抑制剂的 X 射线结构已经得到解决，从而提供了一个强大的工具，我们将用它来通过基于结构的药物设计来指导抑制剂的细化过程。该项目的目标是开发一种小分子 LF 抑制剂来治疗炭疽病，在第一阶段，我们将应用成熟的药物和组合化学技术；抑制剂-酶复合物 X 射线晶体学和基于结构的药物设计 (SBDD) 来快速治疗炭疽病。合成和评估衍生物这个新的经过验证的热门系列。在迭代过程中，我们将通过测量衍生物的酶活性、细胞活性和特异性来探索重点化合物库，以寻找有助于更紧密结合和更有效抑制 LF 的特征。我们将确定与 LF 活性位点结合的改进抑制剂的 X 射线结构，并利用这些数据开发精细的药效团模型，以指导进一步探测结构活性关系 (SAR)。此外，我们将评估化合物的最佳 ADME（吸收、分布、代谢、消除）特性（例如肝微粒体稳定性、Cyp 亚型抑制和 Caco2 通透性）。具有足够酶促、细胞效力和 ADME 特性的化合物将以 1-2 g 水平合成，并在 LF 诱导的大鼠死亡模型中测试功效。成功地将大鼠从 LF 诱导的死亡中拯救出来将使化合物成为体内验证的先导化合物。在第二阶段，我们将进一步评估这些先导药物在两个物种中的体内功效、药代动力学特性、毒性和安全药理学，以便将它们开发成适合人体临床试验的pre-IND临床候选药物（第三阶段）。

项目成果

Small molecule inhibitors of anthrax lethal factor toxin.

炭疽致死因子毒素的小分子抑制剂。

• 发表时间: 2014-01-01
• 影响因子: 3.5
• 作者: Williams, John D;Khan, Atiyya R;Cardinale, Steven C;Butler, Michelle M;Bowlin, Terry L;Peet, Norton P
• 通讯作者: Peet, Norton P

A Survey of Solvents for the Conrad-Limpach Synthesis of 4-Hydroxyquinolones.

4-羟基喹诺酮类药物 Conrad-Limpach 合成溶剂的调查。

• 发表时间: 2009-01-01
• 影响因子: 2.1
• 作者: Brouet JC;Gu S;Peet NP;Williams JD
• 通讯作者: Williams JD