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

描述(申请人提供)：在口腔、颅颌面(CMF)、矫形外科和心血管外科等领域，金属种植体周围的感染是导致种植体失败的常见原因，有时甚至是毁灭性的。这些感染是由于在设备表面建立生物膜而引起的，不仅需要进行新的手术，而且本身就对生命和肢体构成了重大威胁。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。AffinEnergy正在开发被称为界面生物材料(IFBMs)的双功能亲和肽，这种材料可以促进药物、蛋白质和细胞在医疗器械表面的附着和保持。利用噬菌体展示，我们已经鉴定出与多种金属高亲和力结合的特定多肽序列，以及另一组与万古霉素高亲和力结合的特定多肽序列。使用适当的连接物将这些序列合成为单一的IFBM，允许在护理点将万古霉素直接附着到金属植入物的表面。然而，到目前为止，AffinEnergy的药物输送涂层仅限于在多肽分子和抗生素之间安排1对1的相互作用。虽然我们已经用这项使用抗生素的技术观察到了有希望的初步数据，但如果能够增加加载到给定表面的药物的量，我们可能会针对其他药物。因为我们的涂料是由高亲和力的多肽组成的，所以使用一种利用多肽的结构和化学性质的方法来扩展我们的技术将是可取的。因此，我们在这里建议将我们的材料和药物结合肽与组装成更高阶纳米结构的序列偶联。我们的目标是从自组装的亲和肽中生成三维表面涂层，能够将更高浓度的药物结合并保持在设备表面。来自我们初步研究的数据表明，与传统的IFBM相比，能够有限自组装的多肽与材料结合在一起，具有更好的抗化学挑战者的能力。本研究计划试图继续这些研究，开发一种将万古霉素附着在金属上的自组装涂层系统。在目标1中，我们将通过添加不同的自组装肽序列来合成和表征万古霉素和金属结合肽。在目标2中，我们将表征在溶液中和在金属表面上自组装亲和肽的生化和生物物理性质。我们将比较传统的IFBms和新的自组装亲和多肽对万古霉素在金属表面的负载密度、载药率和释放率的影响。最后，在目标3中，我们将表征万古霉素与传统的IFBM和自组装亲和多肽连接到金属上的抗微生物活性。这些目标的成功实现将鼓励我们利用更多的亲和药物、蛋白质、细胞和材料亲和多肽的组合，将自组装亲和多肽扩展到其他临床应用。 公共卫生相关性：金属五金周围的感染是口腔、颅颌面(CMF)、矫形外科和心血管外科等多个医疗领域种植失败的常见原因，有时甚至是毁灭性的。由于在设备表面建立了致病生物膜，这些感染不仅需要进行新的手术，而且本身就对生命和肢体构成了重大威胁。建立在金属硬件上的生物膜细菌基本上不可能通过任何方法根除，除非植入法。降低与金属植入物相关的感染率的方法显然将使社会受益。我们建议开发一种自组装的亲和肽涂层，该涂层将促进抗生素在护理点附着到广泛的金属植入物上，以减少其表面的微生物定植。

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.