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

• 批准号: 7320472
• 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
• 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/7320472
• 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

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（申请人提供）：基质细胞产生的信号分子参与肿瘤转化，可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)是由存在于上皮癌部位的反应性基质细胞选择性产生的，但在任何其他成人组织的基质中不表达。FAP是一种膜结合丝氨酸蛋白酶，具有二肽基肽酶、明胶酶和胶原酶活性。因此，本研究的目的是确定通过FAP激活的前药选择性消除肿瘤相关基质细胞是否能有效地靶向治疗癌症。为了实现这一目标，提出了以下具体目标：(1)定义蛋白水解活性FAP的选择性肽底物；(2)通过将FAP肽与高效的thapsigarin类似物偶联，合成FAP可切割的前药；(3)评价fap活化前药的体内药理学、毒理学及抗肿瘤效果。为了实现目标1，我们生成了重组FAP，并生成了重组胶原蛋白中切割位点的图谱。基于这些切割位点的肽将被评估为推测的FAP底物。此外，我们已经生成了一个随机噬菌体底物库，以评估更多样化的肽作为FAP底物的集合。在Aim 2中，我们将从这些FAP底物中选择最好的来生产FAP激活的thapsigargin前药，这些前药将被表征为FAP在人和小鼠血浆中的水解和稳定性。此外，将测试前药对一组FAP阴性的癌细胞系和一组FAP转染的癌细胞系的毒性。被FAP有效水解、在血浆中稳定、对FAP阴性细胞系毒性最小的前药将在Aim 3中进一步进行体内评估，以确定对产生一系列基质的人类癌症异种移植物的疗效。这些异种移植物将以基质的百分比、FAP的产生和酶活性为特征。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。本提案中描述的研究将定义针对支持结构(即。“基质”)而不是癌细胞本身可以是有效的治疗方法。因此，FAP激活的前药可能代表一种新的针对多种人类癌症的靶向治疗方法。