Chemotherapy targeting human thymidylate synthase messenger RNA

针对人胸苷酸合酶信使 RNA 的化疗

基本信息

批准号：
7844811
负责人：
Thomas C Hermann
金额：
$ 32.53万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
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/7844811
关键词：
Affinity Antineoplastic Agents Binding Biochemical Biological Biological Assay Biological Factors Breast 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 novel piperidine prevent public health relevance research study scaffold small molecule three dimensional structure thymidylate translation assay

项目摘要

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（TS）表达的新型抗癌药物，胸苷酸合酶是 DNA 生物合成的唯一细胞内胸苷酸从头来源。具体来说，我们想要合成和鉴定能够选择性结合和稳定 TS mRNA 5' 非翻译区 (NTR) 中的结构化调节元件的化合物，该元件包含隔离在 RNA 发夹元件中的翻译起始密码子。通过阻止细胞翻译机器进入起始位点，TS mRNA 结合化合物将模仿 TS 蛋白的作用，TS 蛋白充当其自身 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 作为翻译抑制的靶点。在拟议的研究中，我们将建立一个迭代过程，旨在发现先导化合物，用于开发新型癌症疗法，与现有的抗癌药物联合使用，这些抗癌药物以该酶为目标，但由于 TS 表达上调而产生耐药性。该项目的具体目标是：1）基于两类专门用于RNA结合的杂环，设计和合成新型RNA偏向配体； 2) 设计并验证代表 TS mRNA 5'-NTR 中调节 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中调控RNA元件的三维结构； 4) 开发 TS mRNA 的 RNA 亲和力和靶点特异性测定，并在新合成的 RNA 偏向配体和已知的 RNA 结合天然产物中鉴定小分子结合物； 5) 评估 TS mRNA 结合剂在结合和体外翻译测定中的特异性靶标识别； 6) 在人类细胞系中测试TS特异性翻译抑制剂，以确定膜通透性、对TS表达的干扰和抗增殖活性； 7)通过X射线晶体学确定TS mRNA-配体复合物的三维结构； 8) 利用生物活性数据和结构信息（结构活性关系，SAR）设计具有潜在改善的结合亲和力的修饰配体。公共健康相关性：这项研究旨在发现新型抗癌疗法，这将大大提高现有药物治疗耐药性乳腺癌和结直肠癌的疗效。