Development of a novel broad spectrum antifungal therapeutic targeting Glycosylphosphatidylinositol (GPI) biosynthesis and cell wall biogenesis

念珠菌和曲霉属中广泛存在唑类耐药。沿着出现棘白菌素 C.抗性glabrata和C.耳感染引起了无法治疗的多药耐药真菌感染的幽灵， 尽管医学取得了进步（癌症化疗、器官移植、早产儿、艾滋病毒/艾滋病治疗） 增加了弱势群体的规模。我们的建议旨在开发一种新的广谱 靶向糖基磷脂酰肌醇（GPI）生物合成以治疗危及生命的抗真菌治疗剂 由于耐药念珠菌和曲霉菌引起的感染，对现有药物无交叉耐药性。我们 目标是： 目标1（第1阶段）。证明通过优化药代动力学（PK）可改善疗效 系列的属性。对M743、M720和水解产物进行代谢鉴别（MetID）研究 缺乏侧链的核心，以识别氧化代谢热点，从而指导有限的铅优化（铅 Opt）努力改善系列的PK特性，同时保持效价、光谱、靶标选择性，以及 最小化细胞毒性。用和在疟原虫的鼠全身感染模型中测试多达2种新的类似物 没有1-氨基苯并三唑（ABT）PK增强剂共同给药。里程碑1.基于MetID研究， 多达30种新的M743类似物。类似物显示IP给药的剂量依赖性降低> 3 log 10， 在有或没有ABT联合给药的鼠疟原虫模型中的真菌负荷（即上级于M720的功效）将 确定该系列的关键（和可解决的）责任，并保证将该计划推进到2期。 目标2（第2阶段）。M743规模放大，Lead Opt和化合物的体外表征。生产M743 规模足以提供基于MetID数据和新兴SAR的完整的Lead Opt工作。表征 与目标1类似，另外强调PK、MOA、降低的血清结合和细胞毒性。里程碑 2.获得3克M743;半合成多达100种新的类似物。识别多达3种具有可接受效力的类似物 和PK（没有ABT共同给药），沿着与经验证的靶点接合，FOR <1 × 109，与 Gwt 1抑制剂，APX 001 A（FICI<0.5），在血清中的进行性活性，和可接受的毒性（体内细胞毒性 vs. HepG 2，体外IC 50>10 μ M，相对于离子通道，CYP，关键PANLABS靶标），以达到目标3。 目标3（第2阶段）。体内表征。将在小鼠中测试多达2种化合物的功效。 肺结核（包括与APX 001共同给药以评估体内协同作用）和侵袭性肺结核（包括与APX 001共同给药以评估体内协同作用） 无ABT联合给药的曲霉病感染模型。里程碑3.半合成200 mg各试验 化合物.证明念珠菌/曲霉属的MIC 90可接受。确定IP的合适制剂 剂量。进行剂量范围研究，以评估较高剂量下的暴露量和耐受性，以指导剂量 选择.在每种感染模型中达到剂量依赖性疗效的最佳类似物（负荷减少> 3个对数 在治疗期间）和总体上有利的药物样性质将被选择作为临床前候选物。

Widespread azole resistance among Candida and Aspergillus spp. along with emerging echinocandin resistance in C. glabrata and C. auris raises the specter of untreatable multidrug resistant fungal infections, even as advances in medicine (cancer chemotherapy, organ transplant, premature infants, HIV/AIDS therapy) have increased the size of the vulnerable population. Our proposal aims to develop a novel broad spectrum antifungal therapeutic targeting Glycosylphosphatidylinositol (GPI) biosynthesis to treat life threatening infections due to drug resistant Candida and Aspergillus with no cross resistance to existing agents. Our Aims are: Aim 1 (Phase 1). Demonstrate improved efficacy is achievable by optimizing pharmacokinetic (PK) properties of the series. Perform metabolic identification (MetID) studies on M743, M720, and the hydrolyzed core lacking the sidechain to identify oxidative metabolic hotspots to guide a limited Lead Optimization (Lead Opt) effort to improve PK properties of the series while maintaining potency, spectrum, target selectivity, and minimizing cytotoxicity. Test up to 2 new analogs in a murine systemic infection model of Candidiasis with and without 1-aminobenzotriazole (ABT) PK enhancer codosing. Milestone 1. Based on MetID studies, synthesize up to 30 new M743 analogs. An analog showing an IP-administered dose-dependent > 3 log10 reduction in fungal burden in a murine Candidiasis model (i.e. superior to M720 efficacy) with or without ABT codosing will identify the key (and addressable) liability of the series and warrant advancement of the program to Ph 2. Aim 2 (Phase 2). M743 scale up, Lead Opt and in vitro characterization of compounds. Produce M743 on scale sufficient to supply a full Lead Opt effort based on MetID data and emerging SAR. Characterize analogs as in Aim 1 with additional emphasis on PK, MOA, reduced serum binding, and cytotoxicity. Milestone 2. Obtain 3g of M743; semisynthesize up to 100 new analogs. Identify up to 3 analogs with acceptable potency and PK (without ABT codosing), along with validated target engagement, FOR <1 x 109, in vitro synergy with Gwt1 inhibitor, APX001A (FICI<0.5), progressible activity in serum, and acceptable toxicity (in vivo cytotoxicity vs. HepG2, in vitro IC50>10 uM vs ion channels, CYPs, critical PANLABS targets) to advance to Aim 3. Aim 3 (Phase 2). In vivo Characterization. Efficacy of up to 2 compounds will be tested in a murine Candidiasis (including codosing with APX001 to evaluate in vivo synergy) and Invasive Pulmonary Aspergillosis infection models without ABT codosing. Milestone 3. Semisynthesize 200 mg of each test compound. Demonstrate acceptable MIC90 across Candida/Aspergillus spp. Identify suitable formulation for IP dosing. Conduct dose-ranging studies to gauge exposures and tolerability at higher doses to guide dose selection. Top analog achieving dose-dependent efficacy in each infection model (> 3 log reduction in burden over therapeutic duration) and overall favorable drug-like properties will be selected as a preclinical candidate.