Discovery of a Platelet Derived Growth Factor Peptide-based Mimetic

发现基于血小板衍生生长因子肽的模拟物

• 批准号: 7210480
• 负责人: PAUL T HAMILTON
• 金额: $ 23.54万
• 依托单位: AFFINERGY ,INC
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2008-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7210480
• 关键词: bioengineering /biomedical engineering; biological signal transduction; biomaterial development /preparation; biomaterial evaluation; biomimetics; biotherapeutic agent; collagen; diabetes mellitus therapy; drug discovery /isolation; foot; growth factor receptors; medical complication; peptide chemical synthesis; peptide library; peptides; phage display; platelet derived growth factor; protein binding; protein sequence; protein structure function; receptor binding; tissue /cell culture; ulcer; wound healing

DESCRIPTION (provided by applicant): Project Summary/Abstract: Diabetic foot ulcers are a significant health problem that imposes high costs on both patients and society (Boulton, Vileikyte et al. 2005). A large proportion of foot ulcers remain unresponsive to available conventional treatment and their associated complications and costs have prompted extensive research that has led to promising breakthroughs. Recombinant growth factors, biodegradable matrices, and bioengineered skin equivalents have been developed to better promote healing (Edmonds, Bates et al. 2000). These therapies have shown promise, but still have limitations including high cost, immunogenicity, and the need for high, repeated doses of growth factor. Platelet Derived Growth Factor (PDGF) is a potent chemotactic, mitogen, and differentiation factor that promotes soft tissue formation and rhPDGF-BB is currently FDA approved to treat diabetic ulcers. Repeated application of high dose PDGF increases the cost and complexity of treatment. In general, biologic therapeutics are more expensive to develop and manufacture than synthetic therapeutics. Therefore, a synthetic alternative that also has a longer resident time could significantly reduce the cost and complexity of current therapies. The goal of this proposal is to generate a PDGF-BB mimetic that can be immobilized on a collagen scaffold used to promote soft tissue and wound healing. We plan to use phage display technology to identify peptides that mimic PDGF-like signaling of the PDGF B-receptor. These peptides will be linked to a previously identified high-affinity, collagen-binding peptide. The resulting hybrid molecule is expected to allow controlled, local delivery of a synthetic molecule from the surface of a collagen matrix. We expect that collagen binding peptides will significantly improve the retention of a bioactive PDGF mimetic in a wound. First, using phage display techniques, we will identify peptides that bind to the PDGF B-receptor. Secondly, to increase the affinity of the peptides for the B-receptors, focused libraries will be constructed based on the sequences of the initial B-receptor-binding peptides and screened again on the B-receptors. Next, synthetic peptides containing 2 copies of the B-receptor binding sequence will be chemically synthesized with a flexible linker and the resulting dimers will be tested for PDGF-BB-like activity with a fibroblast proliferation assay. Finally, we will synthesize combinations of Affinergy's existing collagen binding sequences and the newly identified PDGF-BB mimetic peptide and the hybrid molecule will be tested for binding to collagen and for its ability to induce fibroblast proliferation while bound to a collagen matrix. Project Narrative: Diabetic foot ulcers are a significant health problem that imposes high costs on both patients and society. A large proportion of foot ulcers remain unresponsive to conventional treatment and have prompted research that has led to promising breakthroughs, including recombinant growth factors, biodegradable matrices, and bioengineered skin equivalents. These therapies have shown promise, but still have limitations including high cost, immunogenicity, and the need for high doses of growth factor. In this project, we will develop a Platelet Derived Growth Factor (PDGF) peptide-based mimetic that can be immobilized on a collagen scaffold used to promote wound healing. We plan to use phage display technology to identify peptides that mimic PDGF's biologic activity. In general, biologic therapeutics are more expensive to develop and manufacture than synthetic therapeutics. Therefore, a synthetic alternative that also has a longer resident time could significantly reduce the cost and complexity of current therapies.

描述（由申请人提供）：项目摘要/摘要：糖尿病足溃疡是一个重大的健康问题，对患者和社会都施加了高昂的成本（Boulton，Vileikyte等，2005）。大部分脚步溃疡对可用的常规治疗及其相关的并发症和成本无反应，这引发了广泛的研究，这导致了有希望的突破。已经开发出重组生长因子，可生物降解的矩阵和生物工程的皮肤等效因素，以更好地促进愈合（Edmonds，Bates等，2000）。这些疗法已显示出希望，但仍有局限性，包括高成本，免疫原性以及对高剂量的生长因子的需求。血小板得出的生长因子（PDGF）是一种有效的趋化，有丝分裂原和分化因子，促进软组织形成，而RHPDGF-BB目前已批准用于治疗糖尿病性溃疡。高剂量PDGF的重复应用增加了治疗的成本和复杂性。通常，生物学治疗剂的开发和制造比合成疗法更昂贵。因此，一种居民时间也更长的合成替代方法可以显着降低当前疗法的成本和复杂性。该提案的目的是产生一种PDGF-BB模拟物，该PDGF-BB可以固定在用于促进​​软组织和伤口愈合的胶原蛋白支架上。我们计划使用噬菌体显示技术来识别模仿PDGF B受体的类似PDGF的肽。这些肽将与先前鉴定的高亲和力，胶原结合肽有关。预计所得的杂化分子将允许从胶原基质表面控制合成分子的局部递送。我们预计胶原蛋白结合肽将显着改善伤口中生物活性PDGF的保留。首先，使用噬菌体显示技术，我们将识别与PDGF B受体结合的肽。其次，为了增加肽对B受体的亲和力，将根据初始B受体结合肽的序列构建聚焦的库，并再次在B受体上进行筛选。接下来，将用柔性接头化学合成，其中包含2份B受体结合序列的合成肽，将测试所得的二聚体，以使用成纤维细胞增殖测定法对PDGF-BB样活性进行测试。最后，我们将合成Affinergy现有的胶原蛋白结合序列的组合以及新近鉴定的PDGF-BB模拟肽和混合分子的结合，以结合与胶原蛋白的结合，并能够诱导成纤维细胞增殖，同时与胶原基质结合。项目叙述：糖尿病足溃疡是一个重大的健康问题，对患者和社会都施加了高昂的成本。大部分的脚溃疡对常规治疗仍然没有反应，并促使研究导致了有希望的突破，包括重组生长因子，可生物降解的矩阵和生物工程的皮肤当量。这些疗法已显示出希望，但仍有局限性，包括高成本，免疫原性以及对高剂量生长因子的需求。在这个项目中，我们将开发基于肽的基于肽的血小板生长因子（PDGF），该因子可以固定在用于促进​​伤口愈合的胶原蛋白支架上。我们计划使用噬菌体显示技术来识别模仿PDGF生物学活性的肽。通常，生物学治疗剂的开发和制造比合成疗法更昂贵。因此，一种居民时间也更长的合成替代方法可以显着降低当前疗法的成本和复杂性。