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：
8710117
负责人：
Aimee L Edinger
金额：
$ 14.34万
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 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8710117
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) 口服治疗药物。在用于治疗 MS 的剂量下，FTY720 对 1-磷酸鞘氨醇受体的作用导致二级淋巴器官中淋巴细胞的隔离。许多研究小组已经证明，在更高剂量下，FTY720 是一种选择性且有效的抗肿瘤药物，可对抗多种肿瘤类型。在小鼠癌症模型中，5-10 mg/kg 剂量的 FTY720 可抑制原发肿瘤和转移瘤的生长，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 FTY720 无法在人类癌症患者中进行测试，因为它在达到抗肿瘤剂量之前会通过激活 S1P 受体 1 和 3 来引发严重的心动过缓。然而，我们最近证明 FTY720 的抗肿瘤作用是通过 S1P 受体独立机制发生的，其中包括营养转运蛋白下调。虽然无法激活 S1P 受体的 FTY720 类似物具有良好的抗肿瘤活性和治疗潜力，但 FTY720 在癌症模型中的分子靶点仍然未知。在该提案中，我们将使用化学生物学方法来识别癌细胞中FTY720及其抗肿瘤类似物的靶点。该项目有两个具体目标：1) 从在完整细胞中具有活性的 FTY720 类似物生成光亲和标记探针，2) 使用一系列阴性对照分离与探针特异性相互作用的蛋白质，以识别非特异性结合伴侣。这些探针将包括二氮丙啶以允许与靶标进行紫外线交联，以及炔烃以通过“点击”化学将生物素共价偶联至探针-靶标复合物。确定癌症模型中 FTY720 及其类似物的分子靶标是开发这种新型癌症治疗方法的关键一步，因为它将使我们能够：1) 定义对这种治疗方法敏感的肿瘤的分子特征，以便药物可以在正确的患者群体中进行测试，2）揭示这些药物的潜在耐药机制，3）预测潜在的毒性，4）对可能具有协同作用的药物组合进行预测，5）测试我们的模型这些化合物使用标准分子和生化技术选择性地杀死癌细胞。