Targeting Oral Biofilms with 2-Aminoimidazole/Triazole Conjugates

描述(申请人提供)：龋齿是美国最流行的疾病之一，85%的成年人和大多数儿童至少接受过一次治疗。虽然这种疾病影响所有人，但对于那些经济地位较低、获得专业口腔护理机会有限的人来说，这尤其是一个问题。龋齿是由于口腔表面生物膜或菌斑中存在产酸菌变形链球菌所致。生物膜是被保护性基质包围的表面附着的细菌群落。生物膜中的细菌对目前使用的抗菌剂的抗药性是自由漂浮细菌的1000倍以上。因此，口腔漱口水中使用的抗菌剂不会 有效消除生物被膜；因此，菌斑不断堆积，导致龋齿。基于这种疾病的患病率和经济差距，需要一种负担得起的、非处方药(OTC)、广泛分布的斑块去除解决方案。在第一阶段，雅居乐科学公司确定了五种能够有效分散口腔生物膜的先导抗生物膜分子 会导致龋齿的物质。这些“Agilyte”分子是从一种控制海洋生物生物污垢的天然产品中提取的。先导化合物能够选择性地抑制和分散变形链球菌的生物被膜，而不影响共生的链球菌种戈登链球菌和血链球菌。在模拟口腔环境中唾液流动的生长条件下，复合药效被证明。这些先导化合物在活性浓度下对细菌无杀伤性，对真核细胞无细胞毒性。这些化合物还能够将人类唾液形成的多种生物膜分散在牙齿状表面。用目前的OTC口腔漱口水对唾液生物膜进行处理的结果表明，AgilyteTM化合物能够将活性成分的功效提高2倍，在更能代表口腔环境的流动条件下，这种提高口腔漱口活性的能力是7倍。在这个第二阶段的项目中，我们将使用体外和体内分析来进一步开发这些先导化合物作为非处方药口腔漱口水的抗生物被膜添加剂。在目标1中，将确定先导化合物的毒性分布。在目标2中，口腔定植的体内模型将被用来确定在体外看到的生物被膜减少能力是否转化为在体内降低制剂中的菌斑负荷的能力。目标2中确定的单一先导化合物将在目标3中进行稳定性评估，并将为大规模生产进行合成优化。这一二期工程的主要里程碑将是鉴定一种无毒的、在体内具有活性的铅分子，并可以大规模合成用于商业生产。在第三阶段，配制的Agilyte“口腔漱口水将接受额外的临床前和临床测试，这将促进一种高效去除牙齿病原体生物膜的非处方药口腔漱口产品的商业化。这种产品有可能改善个人的口腔保健，从而降低龋齿的发生率，特别是对于那些受这种疾病影响最大的低经济地位的人。 公共卫生相关性：龋齿发病率惊人，发病率仅次于普通感冒。龋齿是由菌斑内的病原菌引起的，菌斑是一种被称为生物膜的混合细菌群落，很难从牙齿和口腔组织表面去除。该项目涉及在口腔漱口水中加入强大的抗生物膜分子，以有效地清除和杀灭菌斑内的细菌，从而降低龋齿的发生率。

DESCRIPTION (provided by applicant): Dental caries is one of the most prevalent diseases in the U.S. with 85% of adults and the majority of children having been treated for at least one incidence. While this disease impacts all individuals, it is especially an issue for those of low economic status who have limited access to professional oral care. Dental caries is attributed to the presence of the acidogenic bacterium Streptococcus mutans within biofilms, or plaque, on oral surfaces. Biofilms are surface-attached communities of bacteria that are surrounded by a protective matrix. Bacteria in biofilms are upwards of 1000 times more resistant to currently used antimicrobials than free-floating bacteria. As such, antimicrobials used in oral rinses do not effectively eliminate biofilms; therefore, plaque continues to accumulate, resulting in dental caries. Based on the prevalence and economic disparity of the disease, an affordable, over-the-counter (OTC), widely-distributed solution to plaque removal is needed. In Phase I, Agile Sciences identified five lead anti-biofilm molecules that are effective at dispersing oral biofilms that cause dental caries. These "Agilyte"" molecules are derived from a natural product that controls biofouling of a marine organism. The lead compounds are able to selectively inhibit and disperse biofilms of S. mutans, while not affecting commensal Streptococcal species, S. gordonii and S. sanguinis. Compound efficacy was demonstrated under growth conditions that mimic saliva flow within the oral environment. The lead compounds are non-cidal to bacteria and not cytotoxic to eukaryotic cells at active concentrations. The compounds are also able to disperse multi-species biofilms formed by human saliva on a tooth-like surface. Treatments of saliva-derived biofilms with current OTC oral rinses revealed that the AgilyteTM compounds are able to increase the efficacy of the active ingredients by >2-fold, and this enhancement of oral rinse activity is seven times greater under flow conditions that are more representative of the oral environment. In this Phase II project, we will use in vitro and in vivo analyses to further develop these lead compounds as anti-biofilm additives for OTC oral rinses. In Aim 1, toxicity profiles of the lead compounds will be determined. In Aim 2, an in vivo model of oral colonization will be used to determine if the biofilm reduction capabilities seen in vitro translat to the ability to decrease plaque load in vivo within formulation. A single lead compound identified in Aim 2 will be evaluated in Aim 3 for stability within formulation and will undergo synthetic optimization for large- scale production. The major milestone to be achieved in this Phase II project will be identification of a lead molecule that is non-toxic, active in vivo, and cn be synthesized on a large scale for commercial production. In Phase III, the formulated Agilyte" oral rinse will undergo additional preclinical and clinical testing that will facilitate commercialization of an OTC oral rinse product that is highly effective at removing pathogenic biofilms from teeth. This product has the potential to improve personal oral healthcare, thereby decreasing the incidence of dental caries, particularly for those of low economic status that are most affected by the disease. PUBLIC HEALTH RELEVANCE: The incidence of dental caries is staggering with occurrence of the disease being second only to the common cold. Dental caries are caused by pathogenic bacteria within plaque, a mixed community of bacteria known as a biofilm that is difficult to remove from tooth and oral tissue surfaces. This project involves incorporating potent anti-biofilm molecules into oral rinses to effectively remove and kill bacteria within plaque so that th incidence of dental caries will be reduced.