Chemotherapy targeting human thymidylate synthase messenger RNA

针对人胸苷酸合酶信使 RNA 的化疗

• 批准号: 8249765
• 负责人: Thomas C Hermann
• 金额: $ 31.55万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8249765
• 关键词: Affinity; Antineoplastic Agents; Binding; Biochemical; Biological; Biological Assay; Biological Factors; Cell Line; Cell Membrane Permeability; Chemicals; Colorectal Cancer; Complex; Crystallization; DNA biosynthesis; Data; Development; Drug Delivery Systems; Elements; Enzymes; Genome; Human; Human Cell Line; Hydroxyl Radical; In Vitro; Indium; Initiator Codon; Lead; Ligand Binding; Ligands; Link; Malignant Neoplasms; Messenger RNA; Modeling; Oligonucleotides; Pharmaceutical Preparations; Process; Production; Proteins; RNA; RNA Binding; RNA library; Regulatory Element; Research; Resistance; Resistance development; Series; Site; Source; Specificity; Structure; Structure-Activity Relationship; Testing; Therapeutic; Therapeutic Uses; Thymidylate Synthase; Thymidylate Synthase Inhibitor; Translations; Up-Regulation; X-Ray Crystallography; anti-cancer therapeutic; base; chemotherapy; design; functional group; improved; in vitro Model; in vivo; inhibitor/antagonist; malignant breast neoplasm; novel; piperidine; prevent; public health relevance; research study; scaffold; small molecule; three dimensional structure; thymidylate; translation assay

DESCRIPTION (provided by applicant): The proposed research is intended to discover novel anti-cancer agents that inhibit expression of thymidylate synthase (TS), the sole intracellular de novo source of thymidylate for DNA biosynthesis. Specifically, we want to synthesize and identify compounds that selectively bind and stabilize a structured regulatory element in the 5' nontranslated region (NTR) of the TS mRNA which contains the translation start codon sequestered in an RNA hairpin element. By preventing access of the cellular translation machinery to the initiation site, TS mRNA-binding compounds would mimic the action of TS protein which acts as a ligand and repressor of its own mRNA. In preliminary experiments we have validated the TS mRNA as a target for translation inhibition via stabilization of the mRNA secondary structure. In the proposed research we will establish an iterative process aiming at the discovery of lead compounds for the development of novel cancer therapeutics for use in combination with existing anti-cancer drugs that target the enzyme but suffer from resistance development via upregulation of TS expression. The specific aims of this project are to: 1) design and synthesize novel RNA-biased ligands based on two classes of heterocycles privileged for RNA binding; 2) design and validate model oligonucleotides representing the regulatory RNA element in the 5'-NTR of TS mRNA for ligand binding studies; 3) determine the three- dimensional structure of the regulatory RNA element in the TS 5'-NTR by X-ray crystallography; 4) develop RNA affinity and target specificity assays for the TS mRNA and identify small molecule binders among the newly synthesized RNA-biased ligands and known RNA-binding natural products; 5) evaluate TS mRNA binders for specific target recognition in binding and in vitro translation assays; 6) test TS-specific translation inhibitors in human cell lines to determine membrane permeability, interference with TS expression and antiproliferative activity; 7) determine the three-dimensional structure of TS mRNA-ligand complexes by X-ray crystallography; 8) design modified ligands with potentially improved binding affinity by using biological activity data and structural information (structure activity relationships, SAR). PUBLIC HEALTH RELEVANCE: This research is aimed at the discovery of novel anti-cancer therapeutics which will greatly increase the efficacy of existing drugs for the treatment of resistant breast and colorectal cancer.

描述（由申请人提供）：拟议的研究旨在发现抑制胸苷酸合酶表达（TS）的新型抗癌药，这是胸中胸甲基甲基甲酯的唯一NOVO来源，用于DNA生物合成。具体而言，我们希望合成和识别在TS mRNA的5'非翻译区（NTR）中选择性结合和稳定结构化调节元件的化合物，该元件包含在RNA发夹元件中隔离的翻译起始密码子。通过防止细胞翻译机械进入起始位点，TS mRNA结合化合物将模仿TS蛋白的作用，TS蛋白充当其自己的mRNA的配体和阻遏物。在初步实验中，我们通过稳定mRNA二级结构验证了TS mRNA作为翻译抑制的靶标。在拟议的研究中，我们将建立一个迭代过程，旨在发现铅化合物开发新型癌症治疗剂，以与现有的抗癌药物结合使用，该抗癌药物针对酶，但通过上调TS表达而遭受耐药性的发展。该项目的具体目的是：1）基于两类用于RNA结合的杂环的设计和合成新型RNA偏置的配体； 2）在TS mRNA的5'-NTR中，设计和验证代表调节RNA元件的模型寡核苷酸用于配体结合研究； 3）通过X射线晶体学确定TS 5'-NTR中调节RNA元件的三维结构； 4）开发了TS mRNA的RNA亲和力和靶特异性测定，并鉴定新合成的RNA偏置配体和已知的RNA结合天然产物之间的小分子粘合剂； 5）评估TS mRNA粘合剂在结合和体外翻译测定中的特定靶标识别； 6）在人细胞系中测试TS特异性翻译抑制剂，以确定膜的渗透性，干扰TS表达和抗增殖活性； 7）通过X射线晶体学确定TS mRNA-配体复合物的三维结构； 8）设计通过使用生物学活动数据和结构信息（结构活动关系，SAR）来设计具有潜在改善结合亲和力的修饰配体。 公共卫生相关性：这项研究旨在发现新型的抗癌治疗剂，这将大大提高现有药物治疗耐药性乳腺癌和结直肠癌的功效。