Light-responsive ruthenium compounds for applications in disease

描述（由申请人提供）：该提案旨在开发一类新的光响应无机材料作为化疗剂。顺铂及其类似物仍然是各种癌症的标准疗法，尽管它们与核酸的非特异性作用机制导致一般细胞毒性和使人衰弱的副作用。将提供几个关键优点的有希望的方法是将金属基药物的反应性与光动力疗法（PDT）的选择性组合以增加恶性组织的靶向并减少副作用。我们最近已经表明，结构畸变可用于促进光化学反应，以产生在基于细胞的测定中具有强亲电性和高度光毒性的反应性钌物质。我们假设，畸变可用于控制光激活的细胞毒性，以产生选择性和有效的化疗药物。我们提出了一种新的类基于铼的化合物，其具有易于修改的模块化设计，以促进关键分子组分的快速掺入，从而有效地开发具有选择性和受控反应性的材料。配位化学将被用来产生一个结构多样的家庭的三维手性钌化合物在自组装的方式。将使用高通量细胞存活测定将材料中的结构变化与细胞毒性功效相关联，并且将评估最有希望的化合物的药代动力学（PK）特性、最大耐受剂量（MTD）和在肺癌和黑色素瘤的小鼠异种移植模型中的功效。作用机制研究将用于确定目前未知的细胞杀伤过程。为了实现这一目标，提出了一种新的化学标记策略，以便于使用电子显微镜成像的活性化合物的亚细胞和潜在的亚细胞器定位。如果成功验证，这种方法可以扩展到组织中的其他成像应用。替代方法利用生物素标记的钌材料用于靶下拉，通过凝胶电泳、色谱法和质谱法的过程鉴定细胞靶。体外和细胞内转录和翻译测定将支持这些发现。在所有作用机制研究中，将使用基准无机化疗药物顺铂进行比较。目标是确定所提出的钌材料是否利用与顺铂相同的生物靶点，或者是否可以通过结构修饰针对不同的功能靶点。在完成这些研究后，我们会 扩展了我们对钌络合物的基本光化学的知识，开发了有效的和选择性的细胞毒性化学实体，确定了它们的作用机制，并开发了一种用于生物系统中小分子成像的新工具。1

DESCRIPTION (provided by applicant): This proposal seeks to develop a new class of light-responsive inorganic materials as chemotherapeutic agents. Cisplatin and its analogues remain the standard therapy for a variety of cancers, despite their nonspecific mechanism of action with nucleic acids, resulting in general cytotoxicity and debilitating side-effects. A promising approach that will provide several key advantages is to combine the reactivity of metal- based drugs with the selectivity of photodynamic therapy (PDT) to increase the targeting of malignant tissues and reducing side-effects. We have recently shown that structural distortion can be utilized to promote photochemical reactions to create reactive ruthenium species that are strongly electrophilic and highly photo-toxic in cell based assays. We hypothesize that distortion can be used to control light-activated cytotoxicity for the creation of selective and potent chemotherapeutics. We propose a new class of ruthenium-based compounds that have a readily modifiable modular design to facilitate rapidly incorporation of key molecular components to efficiently develop materials exhibiting selectivity and controlled reactivity. Coordination chemistry will be used to generate a structurally diverse family of three-dimensional chiral ruthenium compounds in a self-assembled manner. The structural variations in the materials will be correlated to cytotoxic efficacy using high-throughput cell survival assays, and the most promising compounds will be assessed for Pharmacokinetic (PK) properties, Maximum Tolerated Dose (MTD) and efficacy in mouse xenograft models of lung cancer and melanoma. Mechanism of action studies will be used to define the currently unknown process of cell killing. In order to accomplish this aim, a novel chemical labeling strategy is proposed to facilitate imaging of the subcellular and potentially sub-organellar localization of active compounds using Electron Microscopy. If successfully validated, this approach could be extended to additional imaging applications in tissues. Alternative approaches utilize biotin-labeled ruthenium materials for target pull-down, with cellular targets identified through a process of gel electrophoresis, chromatography, and mass spectrometry. In vitro and in cell transcription and translation assays will support these findings. Cisplatin, the benchmark inorganic chemotherapeutic, will be used for comparison in all mechanism of action studies. The goal is to determine if the proposed ruthenium materials utilize the same biological target(s) as cisplatin, or may be directed to different functional targets via structural modification. Upon completion of these studies, we will have expanded our knowledge of the fundamental photochemistry of ruthenium complexes, developed potent and selective cytotoxic chemical entities, identified their mechanism of action, and developed a new tool for the imaging of small molecules in biological systems. 1