Self Assembling High Affinity Peptides for Point of Care Drug-Device Combinations

描述（由申请人提供）：金属植入物周围的感染是许多领域（包括口腔、颅颌面（CMF）、骨科和心血管手术）中植入物失效的常见原因，有时甚至是破坏性原因。这些感染是由于在器械表面建立生物膜而引起的，不仅需要进行新的手术，而且本身对生命和肢体构成重大威胁。减少金属植入物微生物定植的新技术将减少这些感染，减轻其严重后果。Affinergy正在开发双功能亲和肽，称为界面生物材料（IFBM），可促进药物，蛋白质和细胞在医疗器械表面的附着和保留。使用噬菌体展示，我们已经确定了特定的肽序列，以高亲和力结合到一些金属和另一组特定的肽序列，以高亲和力结合到万古霉素。使用适当的接头将这些序列合成为单个IFBM允许万古霉素在即时护理时直接连接到金属植入物的表面。然而，到目前为止，Affinergy的药物递送涂层仅限于在肽分子和抗生素之间安排1对1的相互作用。虽然我们已经观察到使用抗生素的这种技术的有希望的初步数据，但如果加载到给定表面的药物量可以增加，我们可能会靶向其他药物。由于我们的涂层由高亲和力肽组成，因此使用利用肽的结构和化学性质的方法来扩展我们的技术将是期望的。因此，我们在这里提出我们的材料和药物结合肽的耦合到组装成更高阶的纳米结构的序列。我们的目标是从自组装亲和肽产生三维表面涂层，能够将显著更高浓度的药物结合并保留到装置表面。来自我们的初步研究的数据表明，与传统的IFBM相比，能够有限自组装的肽与对化学挑战者具有改善的抗性的材料结合。 本研究项目试图继续这些研究，以开发一种自组装涂层系统，将万古霉素附着在金属上。在目标1中，我们将合成和表征万古霉素和金属结合肽，并添加各种自组装肽序列。在目标2中，我们将表征溶液中和金属表面上的自组装亲和肽的生物化学和生物物理性质。 我们将比较负载密度，结合率，和万古霉素从金属涂层与传统的IFBM和新的自组装亲和肽的释放。最后，在目标3中，我们将用传统IFBM和自组装亲和肽表征与金属连接的万古霉素的抗微生物活性。这些目标的成功完成将鼓励我们利用Affinergy药物、蛋白质、细胞和材料亲和肽的额外组合将自组装亲和肽扩展到其他临床应用。 公共卫生相关性：金属硬件周围的感染是许多医疗领域（包括口腔、颅颌面（CMF）、骨科和心血管外科）中植入物失效的常见原因，有时甚至是破坏性原因。 由于在器械表面上建立致病性生物膜，这些感染不仅需要进行新的手术，而且本身对生命和肢体构成重大威胁。在金属硬件上建立自己的生物膜细菌基本上是不可能通过任何手段消除的，除了去除。降低与金属植入物相关的感染率的方法将明显有益于社会。我们建议开发一种自组装亲和肽涂层，该涂层将促进抗生素在护理点与各种金属植入物的附着，以减少其表面上的微生物定植。

DESCRIPTION (provided by applicant): Infection surrounding metal implants is a common and sometimes devastating cause of implant failure in a number of fields including oral, craniomaxillofacial (CMF), orthopedic, and cardiovascular surgery. These infections, which arise from the establishment of biofilms on device surfaces, not only necessitate new surgeries but in themselves present a significant threat to life and limb. New technologies that decrease microbial colonization of metal implants would reduce these infections, mitigating their severe consequences. Affinergy is developing bifunctional affinity peptides, called interfacial biomaterials (IFBMs), that promote the attachment and retention of drugs, proteins, and cells on the surface of medical devices. Using phage display, we have identified specific peptide sequences that bind with high affinity to a number of metals and another specific set of peptide sequences that bind with high affinity to vancomycin. Synthesizing these sequences as a single IFBM using appropriate linkers permits the attachment of vancomycin directly to the surface of a metal implant at point-of-care. Thus far however, Affinergy's drug delivery coatings have been limited to arranging a 1-to-1 interaction between a peptide molecule and an antibiotic. While we have observed promising preliminary data with this technique using antibiotics, there are likely additional drugs we might target if the amount of drug loaded to a given surface could be increased. Because our coatings are comprised of high affinity peptides, expanding our technology using an approach which employs the structural and chemical properties of peptides would be desirable. We therefore propose here the coupling of our material and drug-binding peptides to sequences which assemble into higher-order nanostructures. Our goal is to generate a three-dimensional surface coating from self-assembling affinity peptides, capable of binding and retaining significantly higher concentrations of a drug to a device surface. Data from our preliminary studies, suggest that peptides capable of limited self-assembly bind to materials with improved resistance to chemical challengers compared to traditional IFBMs. This research program attempts to continue these studies, toward developing a self assembling coating system, attaching vancomycin to metal. In Aim 1, we will synthesize and characterize vancomycin and metal binding peptides with the addition of various self-assembling peptide sequences. In Aim 2, we will characterize the biochemical and biophysical nature of self-assembling affinity peptides in solution and on metal surfaces. We will compare the loading density, on-rate, and release of vancomycin from metal coated with traditional IFBMs and new self-assembling affinity peptides. Finally, in Aim 3, we will characterize the anti-microbial activity of vancomycin linked to metals with traditional IFBMs and self-assembling affinity peptides. Successful completion of these aims would encourage us to expand self-assembling affinity peptides to other clinical applications utilizing additional combinations of Affinergy drug, protein, cell, and material affinity peptides. Public Health Relevance: Infection surrounding metal hardware is a common and sometimes devastating cause of implant failure in a number of medical fields including oral, craniomaxillofacial (CMF), orthopedic, and cardiovascular surgery. Arising from the establishment of pathogenic biofilms on device surfaces, these infections not only necessitate new surgeries but in themselves present a significant threat to life and limb. The biofilm bacteria that establish themselves on metal hardware are essentially impossible to eradicate by any means except explantation. Methods that decrease infection rates associated with metal implants would clearly benefit society. We propose to develop a self-assembling affinity peptide coating that will promote attachment of antibiotics at point of care to a wide range of metal implants to decrease microbial colonization on their surfaces.