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

在念珠菌和曲霉菌中广泛存在的唑类耐药性。伴随着新兴的Echinocandin 光滑葡萄球菌和金黄色葡萄球菌的耐药性增加了无法治愈的多重耐药真菌感染的幽灵， 即使是医学上的进步(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 以糖基磷脂酰肌醇(GPI)生物合成为靶点的抗真菌治疗 由耐药念珠菌和曲霉菌引起的感染，对现有药物没有交叉耐药性。我们的 目标是： 目标1(阶段1)。证明通过优化药代动力学(PK)可以提高疗效 该系列的属性。对M743、M720及其水解物进行代谢鉴定(MetID)研究 缺乏侧链的核心识别氧化代谢热点，指导有限的铅优化(铅 OPT)努力改善该系列的PK属性，同时保持效力、光谱、靶选择性和 将细胞毒性降至最低。在念珠菌病小鼠全身感染模型中测试多达2个新的类似物 不添加1-氨基苯并三氮唑(ABT)PK增强剂。里程碑1.基于MetID研究，合成 多达30款新的M743模拟器。一种显示IP给药剂量依赖的&gt；3log10减少的模拟 用或不用ABT联合用药的小鼠念珠菌病模型中的真菌负荷(即优于M720疗效) 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PH2。 目标2(阶段2)。M743放大，铅OPT和化合物的体外表征。生产M743 根据MetID数据和新出现的SAR，规模足以提供全面的牵头OPT努力。刻画 与目标1中的类似物，特别强调PK、MOA、降低血清结合和细胞毒性。里程碑 2.获得M743的3G；半合成多达100个新的类似物。识别最多3个效价可接受的类似物 和PK(无ABT配伍)，以及有效的靶点参与，用于1 x 109，体外协同作用 Gwt1抑制剂APX001A(FICI&lt；0.5)，血清中的进展活性和可接受的毒性(体内细胞毒性 VS HepG2，体外IC50和GT；10um vs离子通道，Cyps，关键的PANLABS靶标)前进到目标3。 目标3(阶段2)。活体表征。多达2种化合物的疗效将在小鼠身上进行测试 念珠菌病(包括使用APX001评估体内协同作用)与侵袭性肺 无ABT协同作用的曲霉病感染模型。里程碑3.每次测试半合成200毫克 化合物。在念珠菌/曲霉属中展示可接受的MIC90。确定适用于IP的配方 给药。进行剂量范围研究，以衡量较高剂量下的暴露和耐受性，以指导剂量 选择。在每个感染模型中实现剂量依赖疗效的顶级模拟(&gt；3log减负 治疗期间)和总体良好的类药物特性将被选为临床前候选药物。

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.