Small molecular weight targeted killing agents against a periodontal pathogen

针对牙周病原菌的小分子量靶向杀灭剂

• 批准号: 8583754
• 负责人: MARK j YOUNG
• 金额: $ 18万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-25 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8583754
• 关键词: Affinity; Animal Model; Animals; Antibodies; Bacteria; Bacteriophages; Biological Models; Bone Tissue; Cell Wall; Cells; Chemistry; Chronic; Collection; Communities; Complementarity Determining Regions; Coupled; Development; Disease; Disease Progression; Eligibility Determination; Engineering; Environment; Etiology; Event; Exhibits; Future; Goals; Health; Homeostasis; Homo; Human; Immune; Immune response; Immune system; In Vitro; Infection; Inflammation; Inflammatory; Knowledge; Lead; Length; Life; Ligands; Light; Membrane; Membrane Proteins; Methods; Microbe; Microbial Biofilms; Molecular Biology; Molecular Weight; Monoclonal Antibodies; Montana; Oxygen; Peptides; Periodontal Diseases; Periodontal Pocket; Periodontitis; Phage Display; Photosensitizing Agents; Play; Process; Research; Risk; Role; Scheme; School Dentistry; Series; Staphylococcus aureus; Sterility; Symptoms; System; Techniques; Testing; Universities; Work; antimicrobial; base; bone loss; chemotherapy; dimer; effective therapy; in vivo; killings; meetings; member; microbial community; oral biofilm; pathogen; periopathogen; prototype; public health relevance; receptor; response; tool; virology

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.

描述（由申请人提供）：牙周病提供了了解从免疫稳态到慢性炎症的过程的机会。从生物膜群中消除选定微生物的能力将为确定假定的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潜力提供重要工具。更广泛地说，共生微生物群落在维持许多粘膜系统的健康中发挥着不可或缺的作用，并且需要选择性杀灭剂来有效治疗这些非无菌界面处的粘膜感染。我们成功地用光敏剂靶向牙周病原菌放线菌聚集菌（Aa）的生物膜，并实现了光激活杀灭。我们使用单克隆抗体 (mAb) 作为靶向部分。单克隆抗体大小的生物分子向生物膜的转运受到严重阻碍。目前还没有针对 Aa 的小分子量靶向杀灭剂，并且没有针对任何给定细菌病原体产生小分子量选择性靶向部分的通用方法。我们的目标是填补这两个空白。小分子量药物不仅具有快速转运至牙周袋和Aa生物膜的优点，而且不太可能具有抗原性，更适合常规合成和应用。我们将采用两种方法来生产针对 Aa 的靶向杀灭剂：1）噬菌体展示（具体目标 1）。我们将使用已建立的方案来筛选对三个靶标具有高亲和力的肽：Aa 生物膜、Aa LPS 和 Aa 外膜蛋白。我们将构建与光敏剂 (PS) 缀合的高亲和力肽的同源和异源二聚体对。 2) 抗体CDR二聚体（具体目标2）。使用抗 Aa mAb 集合作为起点，我们将生产由互补决定区 (CDR) 组成的小分子量靶向肽。对 Aa 生物膜表现出高亲和力的 CDR 将通过亲水间隔基偶联并与 PS 缀合。成功完成我们的具体目标将产生一种工具，可用于在动物和人类研究中探讨 Aa 在牙周病进展中的作用，并提供一种生产针对多种牙周病原体的小分子量靶向杀灭剂的方法。