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：
7822908
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/7822908
关键词：
Adult Affect Bacteriophages Binding Biological Biological Factors Breast Cancer cell line Cancerous Carboxypeptidase Carcinoma Cell Line Cell Survival Cells Characteristics Collagen Collection Colorectal Cancer Coupling Cytotoxic agent Desmoplastic Development Dipeptidyl Peptidases Disseminated Malignant Neoplasm Dose Drug Kinetics Endothelial Cells Epithelial Epithelial Cells Epithelial Neoplasms Extracellular Matrix Fibroblasts Gelatinases Gene Expression Glutamate Carboxypeptidase II Goals Growth Growth Factor Human Hydrolysis In Vitro 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 Recombinants Regimen Research Personnel Series Serine Protease Signal Transduction Inhibitor Signaling Molecule Site Stromal Cells Stromal Neoplasm Structure Surface Thapsigargin Time Tissues Toxic effect Toxicity Tests Toxicology Xenograft procedure analog antiangiogenesis therapy antitumor agent base cancer cell cancer therapy cancer type cell killing collagenase cytotoxic cytotoxicity effective therapy 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，并可以直接调节已建立的癌细胞的生长和存活。成纤维细胞活化蛋白α（FAP）是由存在的反应性基质细胞选择性产生的，存在于上皮癌部位，但在任何其他成年组织的基质中均未表达。 FAP是一种膜结合的丝氨酸蛋白酶，具有二肽基肽酶，明胶酶和胶原酶酶活性。因此，该提案的目的是确定通过FAP激活前药选择性消除肿瘤相关的基质细胞是否有效，有针对性地治疗癌症。为了实现这一目标，提出了以下特定目标：（1）定义蛋白水解活性FAP的选择性肽底物；（2）通过将FAP肽与Thapsigargin的高效类似物耦合，以合成FAP可裂化的前药；（3）评估这些FAP激活前药在体内的药理学，毒理学和抗肿瘤功效。为了完成目标1，我们已经生成了重组FAP，并生成了重组胶原蛋白I中的裂解位点图。基于这些裂解位点的肽将被评估为假定的FAP底物。此外，我们已经生成了一个随机的噬菌体底物库，以评估更多样化的肽作为FAP底物。在AIM 2中，我们将选择其中最好的FAP底物来产生FAP活化的Thapsigargin前药，这些前药将以人和小鼠血浆中的FAP水解和稳定性为特征。此外，将测试前药的毒性对FAP阴性癌细胞系和FAP转染线作为阳性对照。通过FAP有效水解，血浆中的稳定以及对FAP阴性细胞系的最小毒性的前药将在体内进一步评估，以确定对人类癌症异种移植物产生一系列基质的功效。这些异种移植物将用于％基质，用于FAP的产生和酶活性。在进行疗效研究之前，将进行药代动力学分析和毒理学研究以确定最佳剂量方案。该提案中描述的研究将定义针对癌症内而不是癌细胞内的辅助结构（即“基质”）的疗法是否可以有效治疗。因此，FAP激活的前药可以代表针对各种人类癌症的新靶向疗法。