Small molecular weight targeted killing agents against a periodontal pathogen

描述(申请人提供)：牙周病提供了一个机会，以了解导致从免疫动态平衡到慢性炎症的过程。从生物膜联合体中消除选定微生物的能力将为确定假定病原体在导致慢性炎症的事件进展中的作用以及共生体减轻这种破坏性免疫反应的可能性提供重要工具。更广泛地说，共生微生物群落在维持一些粘膜系统的健康方面发挥着不可或缺的作用，需要选择性杀伤剂来有效治疗这些非无菌界面上的粘膜感染。我们用光敏剂成功地靶向牙周病原体放线菌伴生菌(Aggregatibacter放线菌伴生菌，Aggregatibacter放线菌，Aggregatibacter放线菌伴生菌，Aggregatibacter放线菌，Aggregatibacter放线杆菌，Aggregatibacter放线菌，Aggregatibacter，Aggregatibacter放线菌)的我们使用了单抗作为靶向部分。单抗大小的生物分子进入生物膜的运输明显受阻。目前还没有针对AA的小分子靶向杀伤剂，也没有通用的方法来制备针对任何给定细菌病原体的小分子选择性靶向部分。我们的目标是填补这两个空白。小分子药物不仅具有快速进入牙周袋和AA生物膜的优点，而且抗原性较小，更适合常规合成和应用。我们将寻求两种方法来生产针对AA的靶向杀伤剂：1)噬菌体展示(特定目标1)。我们将使用已建立的方法来筛选对三个靶点具有高亲和力的多肽：AA生物膜、AA内毒素和AA外膜蛋白。我们将构建连接到光敏剂(PS)上的高亲和力多肽的同源和异源二聚体对。2)抗体CDR二聚体(特异靶2)。以一系列抗AA单抗为起点，我们将制备由互补决定区(CDRs)组成的小分子量靶向多肽。对AA生物膜具有高亲和力的CDR将通过亲水性间隔物偶联到PS上。我们的特定目标的成功完成将产生一种工具，可用于在动物和人类研究中探索AA在牙周病进展中的作用，并提供一种生产针对各种周围病原体的小分子靶向杀伤剂的方法。

DESCRIPTION (provided by applicant): Periodontal disease provides an opportunity to understand the processes that lead from immune homeostasis to chronic inflammation. The ability to eliminate a selected microbe from a biofilm consortium would provide an essential tool for determining the role of putative pathogens in the progression of events that lead to chronic inflammation and the potential of commensals to mitigate this destructive immune response. More broadly, commensal microbial communities play an integral role in maintaining the health of a number of mucosal systems and selective killing agents will be needed for effective treatment of mucosal infections at these non- sterile interfaces. We successfully targeted biofilms of the periodontal pathogen Aggregatibacter actinomycetemcomitans (Aa) with a photosensitizer and obtained light activated killing. We used a monoclonal antibody (mAb) as the targeting moiety. Transport of biomolecules the size of mAb into biofilms is significantly hindered. There are currently no small molecular weight targeted killing agents for Aa, and there are no general methods for producing small molecular weight selective targeting moieties against any given bacterial pathogen. Our objective is to fill both these gaps. Small molecular weight agents will not only have the advantage of rapid transport into periodontal pockets and Aa biofilm, but are less likely to be antigenic and will be more appropriate for routine synthesis and application. We will pursue two approaches for producing targeted killing agents against Aa: 1) Phage display (Specific aim 1). We will use established protocols for screening peptides with high affinity for three targets: Aa biofilm, Aa LPS and an Aa outer membrane protein. We will construct homo and hetero dimer pairs of high affinity peptides conjugated to a photosensitizer (PS). 2) Antibody CDR dimers (Specific aim 2). Using a collection of anti-Aa mAb as a starting point we will produce small molecular weight targeting peptides consisting of complementarity determining regions (CDRs). CDRs exhibiting high affinity to Aa biofilm will be coupled via a hydrophilic spacer and conjugated to a PS. Successful completion of our specific aims will produce a tool that can be used to probe the role of Aa in periodontal disease progression in both animal and human studies and provide an approach for producing small molecular weight targeted killing agents against a variety of periopathogens.