2-adamantanamine Based Therapeutic for Recurrent Oropharyngeal Candidiasis

基于 2-金刚烷胺的复发性口咽念珠菌病治疗

• 批准号: 8060513
• 负责人: Thomas A Dahl
• 金额: $ 29.92万
• 依托单位: ARIETIS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-10-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8060513
• 关键词: Adherence; Amantadine; Antiviral Agents; Candida; Candida albicans; Candidiasis; Cells; Clinical; DNA; Data; Epithelium; Evaluation; Event; Goals; HIV; Human; Hyphae; In Vitro; Individual; Infection; Measures; Miconazole; Modeling; Mycoses; Pathogenesis; Patients; Pharmaceutical Preparations; Phase; Phenotype; Predisposition; Recurrence; Refractory; Relapse; Reporting; Resistance; Rimantadine; Surface; Testing; Therapeutic; Validation; analog; antimicrobial; base; cellular targeting; drug candidate; drug development; drug production; genome sequencing; mutant; novel; oral cavity epithelium; oropharyngeal thrush; overexpression; pathogen; prevent; protein expression; reconstitution; resistance mutation

DESCRIPTION (provided by applicant): The goal of this Phase I project is to evaluate 2-adamantanamine compounds as a potential therapeutic to treat recurrent oropharyngeal candidiasis (OPC) in HIV patients. 2-adamantanamine is a close structural analog of the antivirals amantadine and rimantadine, making it an attractive drug candidate. Refractory OPC is currently reported in 4-5% of HIV infected individuals. Upon adherence to a surface, C. albicans forms invasive hyphae. We found that 2-adamantanamine prevented invasion and hyphal elongation, two key events in Candida pathogenesis. Recalcitrance to antimicrobial treatment is another important feature of OPC. Apart from forming hyphae, attachment leads to production of drug-tolerant persister cells (LaFleur et al., 2006). We find that patients with unresolved OPC carry high-persistent (hip) mutants of C. albicans, apparently selected by antimicrobial therapy. A combination of AC17 and miconazole was able to eradicate C. albicans cells, including persisters in hip strains. We will evaluate the ability of 2-adamantanamine compounds alone and in combination with miconazole to suppress C. albicans in a reconstituted human oral epithelium model of candidiasis. This will serve as a basis for a Phase II drug development project. The Specific Aims of the project are: Aim 1. In vitro validation of 2-adamantanamine compounds. Many 2-adamantanamine analogs are available commercially, facilitating the evaluation of this class of compounds. The efficacy of 2- adamantanamine and its analogs in inhibiting invasion and hyphal elongation of Candida albicans clinical isolates in vitro will be measured. 26 available analogs will be examined, and 3 of the most potent compounds will be tested against a panel of 20 clinical isolates in order to determine the minimal concentration inhibiting hyphal elongation and invasion in 90% of the strains (MHE90). Milestone: Identify three 2-adamantanamine analogs with a satisfactory MHE90. Aim 2. Efficacy of 2-adamantanamine analogs in a reconstituted human oral epithelium model of candidiasis. The efficacy of the selected 2-adamantanamine analogs against Candida will be evaluated in the reconstituted human oral epithelium model. Compounds will be tested prophylactically and as a treatment for established infection alone and in combination with miconazole and clotrimazole. Compound efficacy will be measured by colony count, LDH release by the epithelium, and microscopically. Milestone: For adamantanamine stand-alone studies: significant reduction in infection. In combination studies, significant reduction in pathogen burden compared to miconazole and clotrimazole alone. Aim 3. Target and mechanism of action of 2-adamantanamine. Identification of resistance mutations by whole genome sequencing, and drug profiling with DNA arrays will be used to determine the target of 2- adamantanamine. Once candidate targets are identified, they will be validated by constructing strains with decreased vs. increased expression of the protein in Candida. We expect increased susceptibility of a strain diminished in target expression as compared to a strain overexpressing the target. Milestone: Identify cellular target(s) responsible for preventing invasion and potentiating miconazole in Candida. )

描述（由申请人提供）：该第一阶段项目的目标是评估 2-金刚烷胺化合物作为治疗 HIV 患者复发性口咽念珠菌病 (OPC) 的潜在疗法。 2-金刚烷胺是抗病毒药物金刚烷胺和金刚乙胺的结构类似物，使其成为有吸引力的候选药物。 据报道，目前 4-5% 的 HIV 感染者患有难治性 OPC。一旦粘附到表面，白色念珠菌就会形成侵入性菌丝。我们发现 2-金刚烷胺可防止念珠菌发病机制中的两个关键事件——入侵和菌丝伸长。对抗菌治疗的抵抗力是 OPC 的另一个重要特征。除了形成菌丝之外，附着还导致耐药持久细胞的产生（LaFleur 等人，2006）。我们发现 OPC 未解决的患者携带白色念珠菌高持久性（臀部）突变体，这些突变体显然是通过抗菌治疗选择的。 AC17 和咪康唑的组合能够根除白色念珠菌细胞，包括髋部菌株中的持续存在细胞。我们将评估 2-金刚烷胺化合物单独以及与咪康唑组合在重建的念珠菌病口腔上皮模型中抑制白色念珠菌的能力。这将作为二期药物开发项目的基础。该项目的具体目标是： 目标 1. 2-金刚烷胺化合物的体外验证。许多 2-金刚烷胺类似物可商购，有助于评估此类化合物。将测量2-金刚烷胺及其类似物在体外抑制白色念珠菌临床分离株的侵袭和菌丝伸长的功效。将检查 26 种可用的类似物，并将针对 20 种临床分离株测试 3 种最有效的化合物，以确定抑制 90% 菌株 (MHE90) 菌丝伸长和侵袭的最低浓度。里程碑：鉴定出三种具有令人满意的 MHE90 的 2-金刚烷胺类似物。目标 2. 2-金刚烷胺类似物在念珠菌病重建人口腔上皮模型中的功效。所选2-金刚烷胺类似物对抗念珠菌的功效将在重建的人口腔上皮模型中进行评估。化合物将进行预防性测试，并作为单独和与咪康唑和克霉唑组合治疗已确定的感染的方法。化合物功效将通过菌落计数、上皮释放的 LDH 以及显微镜来测量。里程碑：对于金刚烷胺独立研究：感染显着减少。在联合研究中，与单独使用咪康唑和克霉唑相比，病原体负荷显着减少。目标 3. 2-金刚烷胺的作用靶点和机制。通过全基因组测序鉴定抗性突变，并使用 DNA 阵列进行药物分析，以确定 2-金刚烷胺的靶标。一旦确定了候选靶点，将通过构建念珠菌中蛋白质表达降低与增加的菌株来验证它们。我们预计与过度表达靶标的菌株相比，靶标表达减少的菌株的敏感性增加。里程碑：确定负责预防念珠菌入侵和增强咪康唑作用的细胞靶点。 ）