Oxadiazole Inhibitors of Non-Stop Ribosome Rescue to treat MDR Neisseria gonorrhoeae

摘要 CDC将MDR淋球菌（Ng）列为三种最紧迫的抗生素耐药性之一 美国的威胁。作为一种革兰氏阴性苛养微生物，Ng引起淋病， 在美国流行性传播细菌感染（STI），估计病例数超过80万例 每年。如果不及时治疗，淋病会导致女性盆腔炎，导致输卵管阻塞。 疤痕和不育或可能传播，引起关节和皮肤的表现。一旦很容易治疗，Ng 几乎对每一种用于治疗它的抗生素都产生了耐药性，留下了阿奇霉素（AZM）和 头孢曲松（CTX）是目前唯一可用的治疗选择。重要的是，最近有一组病例 在夏威夷报道，对AZM和CTX都有抗性，突出了对新的抗生素的迫切需要。 靶向抗药性Ng感染的治疗剂。 细菌翻译受到转录错误、mRNA损伤和翻译移码的困扰 这些事件导致不停止的核糖体复合物，阻止蛋白质产物的释放并进一步抑制 翻译.不停止复合物的恢复是由反式翻译介导的关键细菌过程，ArfA 或ArfB作用于在所有测序的细菌基因组中高度保守的细菌核糖体区域。 在初步研究中，我们证明了基于恶二唑的化合物抑制不停止的核糖体拯救， 作为强效抗菌剂，可对抗一系列病原体（包括Ng），MIC 90值可对抗Ng 范围为0.8-1.6 µM（0.25-0.54 µg/mL）。这些化合物对哺乳动物的毒性最小 细胞，良好的药代动力学和体内抗生素活性在F. tularensis小鼠模型。 本提案的目的是优化前导恶二唑抑制剂的不停止核糖体拯救， 一类新的广谱治疗剂，用于抗淋病奈瑟氏菌， 将该化合物推向研究性新药（IND）-使GLP毒理学和安全性 药理学研究和IND前提交。为此，我们将实现七个具体目标 目标：在目标1中，我们将通过SAR驱动的模拟生成优化电极导线系列。在目标2中， 通过体外生物学和ADME评价优先考虑先导系列类似物。在目标3中，我们将确认和 进一步探讨恶二唑类抑制剂在Ng中不停止核糖体拯救的作用机制和特异性。 在目标4中，我们将根据体内特性选择临床前候选药物和备用药物。在目标5中， 开展IND使能药代动力学、毒理学和安全药理学研究。在Aim 6中，我们将执行 CMC研究，针对GMP生产商的1片非GMP原料药。在目标7中，我们将要求 和药监局开了一个新药申请前会议

Abstract The CDC lists MDR Neisseria gonorrhoeae (Ng) as one of the three most urgent antibiotic resistance threats in the United States. A Gram-negative fastidious organism, Ng causes gonorrhea, the second-most prevalent sexually transmitted bacterial infection (STI) with >800,000 estimated cases in the United States annually. Left untreated, gonorrhea can cause pelvic inflammatory disease in women, leading to fallopian tube scarring and infertility or may disseminate, causing joint and skin manifestations. Once easily treatable, Ng has evolved resistance to nearly every antibiotic used to treat it, leaving a combination of azithromycin (AZM) and ceftriaxone (CTX) as the only currently available treatment option. Importantly, a cluster of cases was recently reported in Hawaii that was resistant to both AZM and CTX, highlighting the critical need for new therapeutics targeting antibiotic-resistant Ng infections. Bacterial translation is plagued by transcription errors, mRNA damage, and translational frameshifting events that result in non-stop ribosome complexes, preventing release of protein products and inhibiting further translation. Recovery of non-stop complexes is a crucial bacterial process mediated by trans-translation, ArfA or ArfB acting on a region of the bacterial ribosome highly conserved across all sequenced bacterial genomes. In preliminary studies, we demonstrated that oxadiazole-based compounds inhibit non-stop ribosome rescue, acting as potent antimicrobials against a range of pathogens, including Ng, with MIC90 values against Ng ranging from 0.8-1.6 µM (0.25-0.54 µg/mL). These compounds exhibit minimal toxicity towards mammalian cells, excellent pharmacokinetics and in vivo antibiotic activity in a F. tularensis mouse model. The objective of this proposal is to optimize the lead oxadiazole inhibitor of non-stop ribosome rescue into a novel class of broad-spectrum therapeutic agents for use against Neisseria gonorrhoeae and to advance this compound towards Investigational New Drug (IND)-enabling GLP toxicology and safety pharmacology studies and pre-IND submission. We will accomplish seven specific aims to achieve this objective: In Aim 1 we will optimize the lead series through SAR-driven analog generation. In Aim 2 we will prioritize lead series analogs through in vitro biology and ADME evaluations. In Aim 3 we will confirm and further explore mechanism of action and specificity of oxadiazole inhibitors of non-stop ribosome rescue in Ng. In Aim 4 we will select a preclinical candidate and backup based on in vivo properties. In Aim 5 we will conduct IND-enabling pharmacokinetic, toxicology and safety pharmacology studies. In Aim 6 we will perform CMC studies, targeting 1 non-GMP patch of drug substance at a GMP manufacturer. In Aim 7 we will request a pre-IND meeting with the FDA.