Finding novel platinum(II) complex anti-cancer drugs with reduced ototoxicity

 描述（由申请人提供）：总体目标是找到对癌细胞显示毒性的铂（II）络合物药物，同时避免耳毒性和听力损失。这项研究将在体外和斑马鱼模型中进行。目前的铂类药物如顺铂具有二胺配体并且具有高度耳毒性，即，产生感觉毛细胞和听力损失。一些新的铂（II）络合物具有三胺或杂环配体，其表现出不同于具有二胺配体的络合物的独特的DNA和蛋白质结合性质。西肯塔基州大学（WKU）的Kevin威廉姆斯博士（生物化学）最近发现铂（II）二胺和三胺复合物在与氨基酸和核苷酸靶点反应时的差异。这些性质可导致减少药物副作用，例如耳毒性。Michael Smith博士（神经生物学）最近表明，可以使用两种耳毒性化学物质-氨基糖苷类抗生素庆大霉素和铂基抗癌药物顺铂-在斑马鱼内耳中检查耳毒性。拟议的项目是WKU这两个实验室之间的跨学科合作。该提议具有两个主要目的：1）确定新的铂（II）络合物是否引起对癌细胞的细胞毒性，和2）确定新的铂（II）络合物是否是耳毒性的（即，损害毛细胞并导致听力损失）。在目标1中，我们将使用细胞毒性试验、电感耦合等离子体发射光谱法（ICP-ES）测量铂进入癌细胞核的吸收和DNA结合试验，在六种不同的癌细胞系中测试六种新型化合物的抗癌活性。在目的2中，将在斑马鱼内耳模型中测试在目的1中发现对癌细胞具有细胞毒性的那些化合物的耳毒性。在用铂化合物或缓冲液对照进行显微注射后，将使用听觉诱发电位（AEP）记录来测量斑马鱼听力阈值。然后解剖内耳，并定量感觉上皮细胞中的细胞死亡和毛细胞损失。这项拟议的研究建立了一个框架，以了解这些新型铂（II）络合物对癌细胞和毛细胞的影响，最终目标是为未来开发避免听力损失副作用的新化疗药物的研究提供基础。该提案实现了区域方案的目标：1）支持有价值的研究; 2）使本科生和研究生接触研究; 3）加强非研究密集型大学的研究环境。西肯塔基州大学立志成为肯塔基州最好的综合性公共机构。该项目支持这一使命，通过改善研究环境，并提供本科生和研究生有机会从事生物医学研究，并支持两个PI的职业目标领先的独立资助的研究实验室。

 DESCRIPTION (provided by applicant): The overarching goal is to find platinum(II) complex drugs that show toxicity against cancer cells, while avoiding ototoxicity and hearing loss. This research will be conducted both in vitro, and in vivo in the zebrafish model. Current platinum-based drugs like cisplatin have diamine ligands and are highly ototoxic, i.e., produce sensory hair cell and hearing loss. Some novel platinum(II) complexes have triamine or heterocyclic ligands that exhibit unique DNA and protein binding properties that differ from that of complexes with diamine ligands. Dr. Kevin Williams (Biochemistry) at Western Kentucky University (WKU) has recently found differences in platinum(II) diamine and triamine complexes in their reactions with amino acid and nucleotide targets. These properties may result in reduced drug side effects such as ototoxicity. Dr. Michael Smith (Neurobiology) recently showed that ototoxicity could be examined in the zebrafish inner ear using two ototoxic chemicals- the aminoglycoside antibiotic gentamicin, and the platinum-based anti- cancer drug cisplatin. The proposed project is an interdisciplinary collaboration between these two laboratories at WKU. This proposal has two primary aims: 1) to determine whether novel platinum(II) complexes cause cytotoxicity to cancer cells, and 2) to determine whether the novel platinum(II) complexes are ototoxic (i.e., damage hair cells and cause hearing loss). In aim 1, we will test six novel compounds in six different cancer cell lines for anticancer activity using cytotoxicity assays, inductively coupled plasma-emission spectroscopy (ICP-ES) to measure uptake of platinum into cancer cell nuclei, and DNA binding assays. In aim 2, those compounds that are found to be cytotoxic to cancer cells in aim 1 will be tested for ototoxicity in a zebrafish inner ear model. Following microinjection with either a platinum-compound or buffer control, zebrafish hearing thresholds will be measured using auditory evoked potential (AEP) recording. Then the inner ears will be dissected and cell death and hair cell loss will be quantified in the sensory epithelia. The proposed study establishes a framework for understanding the effects of these novel platinum(II) complexes on cancer cells and hair cells, with the ultimate goal of providing the foundation for future investigations developing new chemotherapy drugs that avoid the side effect of hearing loss. This proposal accomplishes the AREA program objectives of: 1) supporting meritorious research; 2) exposing undergraduate and graduate students to research; and 3) strengthening the research environment in non- research intensive universities. Western Kentucky University aspires to be the best comprehensive public institution in Kentucky. This project supports this mission by enhancing the research environment and by providing undergraduate and graduate students with opportunities to be engaged in biomedical research, and supports both P.I.s' career goals of leading independently-funded research laboratories.