Identification of the anti-neoplastic target of bioactive FTY720 analogs

生物活性 FTY720 类似物抗肿瘤靶点的鉴定

基本信息

批准号：
8568587
负责人：
Aimee L Edinger
金额：
$ 19.45万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
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：
2013-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8568587
关键词：
Affect Affinity Affinity Labels Agonist Alkynes Animal Model Antineoplastic Agents Architecture Award Azides Binding Bioavailable Biochemical Biological Biological Assay Biology Biotin Bradycardia Cancer Model Cancer Patient Cells Ceramides Characteristics Chemicals Chemistry Coat Protein Complex I Complex Copper Coupled Data Development Diazomethane Dose Dose-Limiting Down-Regulation Drug Combinations Drug Targeting Drug resistance Employee Strikes Exhibits FDA approved Funding Growth Human Hydrocarbons In Vitro Ligands Lymphocyte Lymphoid Malignant Neoplasms Mass Spectrum Analysis Modeling Molecular Molecular Target Multiple Sclerosis Mus Neoplasm Metastasis Nutrient Oral Organ Patients Permeability Pharmaceutical Preparations Primary Neoplasm Probability Protein Phosphatase 2A Regulatory Subunit PR53 Proteins Protocols documentation Publishing Pyrrolidines Radiolabeled Reaction Receptor Activation Series Specificity Sphingolipids Sphingomyelins Sphingosine Sphingosine-1-Phosphate Receptor Staging Streptavidin Structure-Activity Relationship Techniques Testing Therapeutic Toxic effect Tumor Suppressor Proteins Water Work affinity labeling analog appendage base cancer cell cancer initiation cancer therapy cancer type catalyst crosslink design effective therapy functional group gel electrophoresis in vivo inhibitor/antagonist killings leukemia neoplastic novel novel strategies patient population prevent protein complex public health relevance pyrrolidine radiotracer success therapeutic development tumor

项目摘要

DESCRIPTION (provided by applicant): SUMMARY FTY720 is FDA-approved oral treatment for multiple sclerosis (MS). At the dose used to treat MS, FTY720's effects on sphingosine-1-phosphate receptors result in the sequestration of lymphocytes in secondary lymphoid organs. At much higher doses, many groups have demonstrated that FTY720 is a selective and effective anti-neoplastic agent that works against a wide variety of tumor types. In mouse cancer models at 5-10 mg/kg, FTY720 inhibits the growth of both primary tumors and metastases while exhibiting minimal toxicity. Despite this striking success in animal models, FTY720 cannot be tested in human cancer patients because it triggers profound bradycardia by activating S1P receptors 1 and 3 before the anti-neoplastic dose is reached. However, we have recently demonstrated that the anti-neoplastic effect of FTY720 occurs through an S1P receptor-independent mechanism that includes nutrient transporter down-regulation. While FTY720 analogs that fail to activate S1P receptors have promising anti-neoplastic activity and therapeutic potential, the molecular target of FTY720 in cancer models remains unknown. In this proposal, we will use a chemical biology approach to identify the target of FTY720 and its anti-neoplastic analogs in cancer cells. This project has two Specific Aims: 1) Generate photo-affinity labeling probes from FTY720 analogs that are active in intact cells, and 2) Isolate proteins that specifically interact with the probes using a series of negative controls to identify non-specific binding partners. These probes will include a diazirine to allow UV-crosslinking to the target and an alkyne to covalently couple a biotin to the probe-target complex via "click" chemistry. Determining the molecular target for FTY720 and its analogs in cancer models is a critical step in the development of this novel approach to cancer therapy because it will allow us to: 1) define the molecular characteristics of tumors that will be susceptible to this therapeutic approach so that drugs can be tested in the correct patient populations, 2) uncover potential mechanisms of resistance to these drugs, 3) anticipate potential toxicities, 4) make predictions about drug combinations that are likely to be synergistic, and 5) test our model for how these compounds selectively kill cancer cells using standard molecular and biochemical techniques.

描述（由申请人提供）：摘要FTY720是FDA批准的多发性硬化症（MS）的口服处理。在用于治疗MS的剂量下，FTY720对鞘氨氨酸1-磷酸受体的影响导致次生淋巴机器人中淋巴细胞的隔离。在更高剂量的情况下，许多组表明FTY720是一种选择性且有效的抗塑性剂，可与多种肿瘤类型作用。在5-10 mg/kg的小鼠癌模型中，FTY720抑制原发性肿瘤和转移的生长，同时表现出最小的毒性。尽管在动物模型中取得了惊人的成功，但在人类癌症患者中无法测试FTY720，因为它通过在达到抗肿瘤剂量之前激活S1P受体1和3来触发深度的心动过缓。但是，我们最近证明，FTY720的抗塑性作用是通过包括营养转运蛋白下调的S1P受体独立机制发生的。尽管未能激活S1P受体的FTY720类似物具有有希望的抗塑性活性和治疗潜力，但癌症模型中FTY720的分子靶标仍然未知。在此提案中，我们将使用一种化学生物学方法来识别癌细胞中FTY720及其抗塑性类似物的靶标。该项目具有两个具体的目的：1）从活性细胞中活跃的FTY720类似物中生成照片亲和力标记探针，以及2）分离蛋白，这些蛋白质使用一系列负面对照来识别非特异性结合伴侣，这些蛋白质特异性地与探针特异性相互作用。这些探针将包括重氮嗪，以使紫外线链接到靶标，并通过“单击”化学反应将生物素与探针靶向复合物共价。 Determining the molecular target for FTY720 and its analogs in cancer models is a critical step in the development of this novel approach to cancer therapy because it will allow us to: 1) define the molecular characteristics of tumors that will be susceptible to this therapeutic approach so that drugs can be tested in the correct patient populations, 2) uncover potential mechanisms of resistance to these drugs, 3) anticipate potential toxicities, 4) make predictions about drug combinations可能是协同作用的，5）测试我们的模型如何使用标准分子和生化技术选择性地杀死癌细胞。