Fibroblast Activation Protein-alpha Activated Anti-Stromal Prodrug Therpay for Ca

成纤维细胞激活蛋白-α 激活抗基质前药治疗 Ca

基本信息

批准号：
7452354
负责人：
Samuel R Denmeade
金额：
$ 31.16万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7452354
关键词：
Adult Affect Appendix Bacteriophages Binding Biological Biological Factors Breast Cancer cell line Cancerous Carboxypeptidase Carcinoma Cell Line Cell Survival Cells Characteristics Class Collagen Collection Colorectal Cancer Coupling Cytotoxic agent Desmoplastic Development Dipeptidyl Peptidases Disseminated Malignant Neoplasm Dose Drug Kinetics Endopeptidases Endothelial Cells Epithelial Epithelial Cells Epithelial Neoplasms Extracellular Matrix Fibroblasts Gelatinases Gene Expression Glutamate Carboxypeptidase II Goals Growth Growth Factor Human Hydrolysis In Vitro Invasive Libraries Liquid substance Lung Lymphocyte Malignant Neoplasms Malignant neoplasm of prostate Maps Mediator of activation protein Membrane Methods Molecular Profiling Morphology Mus Neoplasms in Vascular Tissue Neoplastic Cell Transformation Normal tissue morphology Nutrient Organ Peptide Hydrolases Peptides Pharmacology and Toxicology Plasma Population Heterogeneity Pro-X dipeptidase Prodrugs Production Proliferating Prostate-Specific Antigen Range Rate Recombinants Research Personnel Series Serine Protease Signal Transduction Inhibitor Signaling Molecule Site Standards of Weights and Measures Stromal Cells Stromal Neoplasm Structure Surface Thapsigargin Time Tissues Toxic effect Toxicity Tests Toxicology Treatment Protocols Xenograft procedure analog antiangiogenesis therapy antitumor agent base cancer cell cancer therapy cancer type cell killing collagenase cytotoxic cytotoxicity fibroblast activation protein alpha in vivo killings macrophage neoplastic cell novel peptide I programs response selective expression success therapeutic angiogenesis therapeutic target trait tumor

项目摘要

DESCRIPTION (provided by applicant): Stromal cell production of signaling molecules is involved in neoplastic transformation and can directly regulate growth and survival of established cancer cells. Fibroblast activation protein alpha (FAP) is selectively produced by reactive stromal cells present within sites of epithelial cancers but is not expressed within stroma of any other adult tissues. FAP is a membrane bound serine protease with dipeptidyl peptidase, gelatinase and collagenase enzymatic activities. Therefore, the objective of this proposal is to determine if selective elimination of tumor associated stromal cells by a FAP activated prodrug would be effective, targeted treatment for cancer. To achieve this objective the following Specific Aims are proposed: (1) to define selective peptide substrates for the proteolytic activity FAP; (2) to synthesize FAP-cleavable prodrugs by coupling the FAP peptide to highly potent analogs of thapsigargin; (3) to evaluate pharmacology, toxicology and antitumor efficacy of these FAP-activated prodrugs in vivo. To accomplish Aim 1 we have generated recombinant FAP and generated a map of cleavage sites within recombinant collagen I. Peptides based on these cleavage sites will be evaluated as putative FAP substrates. In addition, we have generated a random phage substrate library to evaluate a more diverse collection of peptides as FAP substrates. In Aim 2, we will select the best of these FAP substrates to produce FAP activated thapsigargin prodrugs which will be characterized for FAP hydrolysis and stability in human and mouse plasma. In addition, the prodrugs will be tested for toxicity against a panel of FAP negative cancer cell lines and a FAP transfected line as a positive control. Prodrugs that are efficiently hydrolyzed by FAP, stable in plasma, and minimally toxic to FAP negative cell lines will be further evaluated in Aim 3 in vivo to determine efficacy against human cancer xenografts producing a range of stroma. These xenografts will be characterized for % stroma, for FAP production and enzymatic activity. Prior to efficacy studies, pharmacokinetic analysis and toxicology studies will be performed to determine optimal dosing regimen. The studies described in this proposal will define whether therapies that target the supporting structures (i.e. "stroma") within cancers rather than the cancer cells themselves can be effective therapy. The FAP- activated prodrug, therefore, could represent a new targeted therapy for a variety of human cancers.

描述（由申请人提供）：信号分子的基质细胞产生参与肿瘤转化并且可以直接调节已建立的癌细胞的生长和存活。成纤维细胞激活蛋白 α (FAP) 由上皮癌部位内的反应性基质细胞选择性产生，但在任何其他成体组织的基质内不表达。 FAP 是一种膜结合丝氨酸蛋白酶，具有二肽基肽酶、明胶酶和胶原酶酶活性。因此，本提案的目的是确定通过 FAP 激活的前药选择性消除肿瘤相关基质细胞是否会是癌症的有效、靶向治疗。为了实现这一目标，提出了以下具体目标： (1) 定义具有蛋白水解活性的 FAP 的选择性肽底物； (2)通过将FAP肽与毒胡萝卜素的高效类似物偶联来合成FAP可裂解的前药； (3)评价这些FAP激活的前药的体内药理学、毒理学和抗肿瘤功效。为了实现目标 1，我们生成了重组 FAP 并生成了重组胶原 I 内的裂解位点图谱。基于这些裂解位点的肽将作为假定的 FAP 底物进行评估。此外，我们还生成了一个随机噬菌体底物库来评估作为 FAP 底物的更多样化的肽集合。在目标 2 中，我们将选择这些 FAP 底物中最好的来生产 FAP 激活的毒胡萝卜素前药，其特征在于 FAP 水解和在人和小鼠血浆中的稳定性。此外，还将测试前药对一组 FAP 阴性癌细胞系和作为阳性对照的 FAP 转染细胞系的毒性。被 FAP 有效水解、在血浆中稳定且对 FAP 阴性细胞系毒性最小的前药将在 Aim 3 体内进一步评估，以确定对产生一系列基质的人类癌症异种移植物的功效。这些异种移植物将表征基质百分比、FAP 产生和酶活性。在功效研究之前，将进行药代动力学分析和毒理学研究以确定最佳给药方案。该提案中描述的研究将确定针对癌症内支持结构（即“基质”）而不是癌细胞本身的疗法是否可以是有效的疗法。因此，FAP 激活的前药可以代表多种人类癌症的新靶向治疗。