Targeting Oral Biofilms with 2-Aminoimidazole/Triazole Conjugates

描述（由申请人提供）：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。虽然这种疾病会影响所有人，但对于那些经济地位低的人来说，这尤其是一个问题，他们无法获得专业的口腔护理。龋齿是由于口腔表面的生物膜或菌斑内存在产酸细菌变形链球菌。生物膜是由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性比自由漂浮的细菌高1000倍以上。因此，用于口腔冲洗的抗菌剂不 有效地消除生物膜;因此，牙菌斑继续积累，导致龋齿。基于该疾病的患病率和经济差异，需要一种负担得起的非处方（OTC）广泛分布的斑块去除解决方案。 在第一阶段，敏捷科学公司确定了五种有效分散口腔生物膜的主要抗生物膜分子 会导致龋齿这些“生物分子”来自一种天然产物，可以控制海洋生物的生物污垢。先导化合物能够选择性地抑制和分散沙门氏菌生物膜。变形链球菌，而不影响链球菌种，S。gordonii和S.吸血鬼在模拟口腔环境内唾液流动的生长条件下证明了化合物功效。先导化合物在活性浓度下对细菌无杀灭作用，对真核细胞无细胞毒性。这些化合物还能够分散由人类唾液在牙齿样表面上形成的多物种生物膜。用目前的OTC口腔冲洗液处理唾液来源的生物膜显示，Escheroyte TM化合物能够将活性成分的功效提高2倍以上，并且在更能代表口腔环境的流动条件下，口腔冲洗活性的增强是7倍。在这个II期项目中，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OTC口腔冲洗液的抗生物膜添加剂。在目标1中，将确定先导化合物的毒性特征。在目的2中，将使用口服定殖的体内模型来确定体外观察到的生物膜减少能力是否等同于制剂内体内减少菌斑负荷的能力。目标2中确定的单一先导化合物将在目标3中评价制剂内的稳定性，并将进行大规模生产的合成优化。在该第二阶段项目中要实现的主要里程碑将是鉴定出无毒的、在体内有活性的并且可以大规模合成用于商业生产的先导分子。在第三阶段，配制的”口腔清洗剂“将进行额外的临床前和临床测试，这将促进OTC口腔清洗剂产品的商业化，该产品在去除牙齿上的致病生物膜方面非常有效。该产品有可能改善个人口腔保健，从而降低龋齿的发病率，特别是对于那些受疾病影响最大的经济地位较低的人。

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.