Purine Synthesis Inhibitors with Selective Folate Receptor Tumor Transport

具有选择性叶酸受体肿瘤转运的嘌呤合成抑制剂

基本信息

批准号：
8613474
负责人：
Charles E. Dann
金额：
$ 54.62万
依托单位：
DUQUESNE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-05 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8613474
关键词：
4-Aminobenzoic Acid 5&apos -AMP-activated protein kinase Affinity Aminopterin Anabolism Antineoplastic Agents Binding Blood Circulation Breast Cell Death Cell Line Cells Cervical Chemotherapy-Oncologic Procedure Chinese Hamster Clinical Clinical Trials Collaborations Coupled Crystallization Development Dose-Limiting Drug Design Drug Transport Drug usage Endometrial Enzymes Equilibrium Evaluation Exhibits Failure Folate Folic Acid Folic Acid Antagonists Glycine Goals Hamsters Hand Human Hydroxymethyltransferases Implant In Situ In Vitro Intracellular Transport Kidney Label Lead Literature Lometrexol Lung Malignant Neoplasms Malignant neoplasm of ovary Measures Metabolic Metabolism Methotrexate Modeling Molecular Molecular Target Non-Small-Cell Lung Carcinoma Normal Cell Nucleosides Nucleotide Biosynthesis Nucleotides Ovarian Ovary Partner in relationship Pathway interactions Patients Pattern Pemetrexed Pharmaceutical Preparations Pharmacology Property Protons Pteridines Purine Nucleotides Purines Radiolabeled Raltitrexed Recombinants Resistance Ribonucleotides Roentgen Rays SCID Mice SKOV3 cells SLC19A1 gene Series Signal Transduction Specificity Staging Structural Biologist Structure System Testing Thiophenes Thymidylate Synthase Toxic effect Treatment Failure Tumor Cell Line analog antitumor agent apical membrane base cancer cell cancer therapy cellular targeting chemotherapeutic agent cytotoxic cytotoxicity design drug development efficacy trial folate-binding protein glycine amide human FRAP1 protein imidazole-4-carboxamide imidazolecarboxamide improved in vivo inhibitor/antagonist killings kinase inhibitor nano neoplastic cell novel overexpression phosphoribosylaminoimidazolecarboxamide formyltransferase polyglutamate public health relevance purine radiotracer receptor expression subcutaneous tumor tumor specificity tumor xenograft uptake

项目摘要

DESCRIPTION (provided by applicant): The failure of cancer chemotherapy can be attributed in large measure to a lack of tumor selectivity of chemotherapeutic agents. Currently approved antitumor antifolates, methotrexate, pemetrexate, raltitrexed and pralatrexate, are all transported by the ubiquitous, reduced folate carrier (RFC) into both tumor and normal host cells, and hence are nonselective for tumor cells and are consequently toxic. If an agent could selectively attack only cancer cells, a qualitative and not just a quantitative difference between cancer cells and normal cells would be at hand for exploitation. We have discovered compounds that specifically target tumor cells by selective transport by folate receptor (FR) -? and -?. These are folic acid transport systems (almost) exclusively expressed by several tumor cells (e.g., ovarian, non-small cell lung cancer, endometrial, renal, breast, cervical, lung) but nt normal cells. Our compounds, once selectively transported into tumors, target the de novo purine nucleotide biosynthesis pathways dependent on folate, i.e. glycinamide ribonucleotide formyltransferase (GARFTase) or 5-amino-4- imidazolecarboxamide ribonucleotide (AICAR) formyltransferase (AICARFTase). These are both novel enzyme targets, as there are no known clinical agents that target de novo purine biosynthesis as their primary mechanism of action. Antipurine antifolates are cytotoxic independent of p53 status. Selectivity of antipurine agents can result from the loss of purine salvage in a large number of human tumors. Inhibition of AICARFTase causes an accumulation of AICAR (ZMP) that, via AMPK, inhibits mTOR, resulting in an additional mechanism of cytotoxic antitumor activity. The goal of this proposal is to optimize our lead structures for tumor specificity via FR? and -? over RFC, and GARFTase or AICARFTase inhibitory activities. In Aim 1, we will synthesize novel analogs from 16 series (two in each series) based on structure-activity profiles of our lead compounds. In Aim 2, we will evaluate compounds from Aim 1 for cytotoxicity in isogenic hamster and human tumor cell line models with established RFC and FR expression, and will identify molecular mechanisms including cellular targets by nucleoside protection, in situ metabolic labeling, analysis of intracellular metabolites, and studies with isolated enzymes. Additional studies will characterize transport properties of the novel analogs with FRs vis ¿ vis RFC, polyglutamate synthesis, and mechanisms of cell death. In Aim 3, we will determine in vivo efficacies of the most potent FR-targeted GARFTase or AICARFTase inhibitors by in vivo toxicity and efficacy trials in FR-expressing human tumor implants in SCID mice. Finally, in Aim 4, we will determine X-ray crystal structures of the most potent and selective analogs with FR? and/or -?, as well as GARFTase or AICARFTase. All the Specific Aims will be performed concurrently from year 1 to year 5. Collectively, our studies will afford a molecular understanding of the interactions of nove analogs with FRs and enzyme targets to guide the design of selective analogs. We anticipate advancing one or more of our novel folate analogs with optimized FR-selective antitumor agents to clinical trials to be used alone or in combination with other agents.

描述（由适用提供）：癌症化学疗法的失败可以很大程度上归因于缺乏化学治疗剂的肿瘤选择性。目前经批准的抗肿瘤抗毒酸酯，甲氨蝶呤，吡甲酸酯，raltrexrex和pralatrexate均由普遍存在的叶酸载体（RFC）转运到肿瘤和正常宿主细胞中，因此，肿瘤细胞不受欢迎，因此是非肿瘤细胞的。如果代理只能选择性地攻击癌细胞，则可以进行定性，而不仅仅是癌细胞和正常细胞之间的定量差异，以进行剥削。我们发现了通过叶酸接收器（FR） - 选择性转运特异性靶向肿瘤细胞的化合物？和 -？。这些是由几个肿瘤细胞（例如卵巢，非小细胞肺癌，子宫内膜，肾脏，乳房，颈椎，肺）独家表达的叶酸转运系统（几乎）。我们的化合物曾经有选择地转运成肿瘤，靶向依赖叶酸的新嘌呤生物合成途径，即乙二胺核糖核苷酸甲基转移酶（Garftase）或5-氨基-4-氨基-4-氨基二唑烷酰胺盒酰胺核苷酸核苷酸核苷酸（aicar）emyltransyltransylansylansylansylansylansransfrase（aicaryltransfrase）。这些都是新型的酶靶标，因为没有已知的临床剂将从头开始的嘌呤生物合成作为其主要作用机理。抗肽抗染料是细胞毒性，与p53状态无关。抗磷脂药物的选择性可能是由于大量人类肿瘤中嘌呤挽救的丧失而导致的。抑制AICARFTase会导致AICAR（ZMP）的积累，通过AMPK抑制MTOR，从而导致了细胞毒性抗肿瘤活性的附加机制。该提案的目的是通过FR优化我们的肿瘤特异性铅结构？和 -？超过RFC，Garftase或AICARFTase抑制活性。在AIM 1中，我们将根据我们的铅化合物的结构 - 活性谱，合成16个系列的新型类似物（每个系列中的两个）。在AIM 2中，我们将评估来自AIM 1的化合物，用于具有已建立的RFC和FR表达的等源性仓鼠和人类肿瘤细胞系模型中的细胞毒性，并将确定分子机制，包括通过核斑点保护，原位代谢标记，分析细胞内代谢物的分析以及与分离的Enzymes的研究。其他研究将表征新型类似物的运输特性，具有VIS RFC，聚谷氨酸合成和细胞死亡的机制。在AIM 3中，我们将通过体内毒性和在SCID小鼠中表达FR的人类肿瘤不屑一顾的体内毒性和有效试验来确定最潜在的FR靶向Garftase或AICARFTase抑制剂的体内效率。最后，在AIM 4中，我们将确定与FR的最潜力和选择性类似物的X射线晶体结构？和/或 - ？，以及garftase或aicarftase。从1年级到5年级，将同时执行所有具体目标。总的来说，我们的研究将对新型类似物与FRS和酶靶标的相互作用有分子的理解，以指导选择性类似物的设计。我们预计，将我们的一种或多种新型的叶酸类似物推进了具有优化的FR选择性抗肿瘤剂的临床试验，可以单独使用或与其他药物结合使用。