DEVELOPMENT OF NOVEL ANTI-CANCER AGENTS BASED ON FUSAROCHROMANONE

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 本研究的主要目的是开发基于镰刀菌色满酮（FC 101 a）的新型抗癌药物，FC 101 a是一种天然真菌毒素和由木贼镰刀菌产生的真菌代谢产物。 FC 101 a由于其多种生物学特性，包括有效的抗血管生成和直接抗肿瘤活性，已被确定为有吸引力的先导候选药物。 与大多数其他生物活性天然化合物一样，FC 101 a的效力在体内会受到影响，该项目的目标是在FC 101 a的一些更有效的类似物之间建立结构-功能关系。 我们在化学和生物合成FC 101 a方面都取得了进展。 我们采用了一种新的途径来合成母体化合物中独特的多官能四碳侧链。 我们还成功地利用Suzuki反应将三氟甲磺酸酯中间体（6-OTf-色满酮和5-氨基-6-OTf-色满酮）偶联到基于苯乙烯的模型侧链上。 13 C和1H-NMR分析证实了两种偶联产物的正确结构。 此外，我们还合成了两个新的FC 101 a的缩醛类似物，并将提出选择这些结构类似物的合理性，为设计这一系列化合物的构效关系。 对于生物合成，我们已经从两个镰刀菌菌株（F4482和F8505）的水稻培养物中获得了类似于FC 101 a的高荧光代谢产物，基于TLC分析。我们还开发了一种高效的HPLC纯化方案，以获得高纯度的该化合物样品。我们目前正在努力扩大这种分离，并通过1H，13 C-NMR以及高分辨率质谱分析来确认这种化合物的结构。 我们已经扩展了我们对FC 101 a生物学功能的初步研究，包括对参与细胞增殖信号通路的关键蛋白的表达水平和磷酸化状态的研究、细胞分选谱和FACS分析。 此外，我们已经启动了几个实验，以评估母体FC 101 a的抗血管生成特性。 最后，我们利用INBRE资金购买或升级了进行这项研究所需的几个关键仪器，并完成了安装和开发适当的学生培训协议，以使用仪器。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The primary goal of this research is to develop novel anti-cancer agents based on fusarochromanone (FC101a), a natural mycotoxin and a fungal metabolite produced by Fusarium equiseti. FC101a has been identified as an attractive lead drug candidate because of its diverse biological properties, including potent anti-angiogenic and direct anti-tumor activity. Like most other bioactive natural compounds, the potency of FC101a is compromised in-vivo, and the project's goal involves the establishment of structure-function relationships among some of FC101a's more potent analogs. We have made progress towards both the chemical and biological synthesis of FC101a. We have employed a new pathway for synthesizing the unique multifunctional four-carbon side chain in the parent compound. We have also been successful in utilizing the Suzuki reaction to couple both triflate intermediates (6-OTf-chromanone and 5-amino-6-OTf-chromanone) to a model side chain based on styrene. 13C and 1H-NMR analyses have confirmed the correct structure for both coupling products. Additionally, we have synthesized two novel acetal analogs of FC101a and will present the rational for choosing these structural analogs for devising structure activity relationships for this series of compounds. For the biological synthesis, we have obtained a highly fluorescent metabolite from the rice culture of two Fusarium strains (F4482 and F8505) that resembles FC101a, based on TLC analyses. We have also developed an efficient HPLC purification protocol to obtain samples of this compound with high purity. We are currently working to scale up this separation and confirm the structure of this compound through 1H, 13C-NMR as well as high-resolution mass spectrometry analyses. We have extended our preliminary studies of the biological function of FC101a, including investigation of the expression levels and phosphorylation states of key proteins involved in cell proliferation signaling pathways, cell sorting profiles, and FACS analyses. In addition, we have initiated several experiments to assess the anti-angiogenic properties of the parent FC101a. Finally, we have utilized the INBRE funding to purchase or upgrade several key pieces of instrumentation needed to conduct this research and have completed the installation and development of appropriate student training protocols for the use of the instrumentation.