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

摘要 CDC将耐多药淋球菌(Ng)列为三种最紧迫的抗生素耐药性之一 在美国的威胁。作为一种革兰氏阴性的挑剔微生物，Ng引起的淋病排在第二位 美国流行的性传播细菌感染(STI)估计有80万例 每年一次。如果不进行治疗，淋病会导致女性盆腔炎，导致输卵管 结疤和不孕，或可能扩散，引起关节和皮肤症状。一旦很容易治疗，Ng就有 对几乎所有用于治疗它的抗生素都产生了进化的耐药性，留下了阿奇霉素(Azm)和 头孢曲松(CTX)是目前唯一可用的治疗方案。重要的是，最近发生了一系列病例 在夏威夷报告了对Azm和CTX都耐药的情况，突显了新的 针对耐药Ng感染的治疗。 细菌翻译受到转录错误、信使核糖核酸损伤和翻译移码的困扰 导致不间断核糖体复合体的事件，阻止蛋白质产物的释放并进一步抑制 翻译。不间断复合体的恢复是一个关键的细菌过程，由反式翻译介导 或ARFB作用于细菌核糖体的一个区域，该区域在所有已测序的细菌基因组中高度保守。 在初步研究中，我们证明了恶二唑类化合物抑制了不间断的核糖体拯救， 作为对包括Ng在内的一系列病原体的有效抗菌剂，对Ng具有MIC90值 范围为0.8-1.6微米(0.25-0.54微克/毫升)。这些化合物对哺乳动物的毒性最小。 细胞，出色的药代动力学和体内抗生素活性在图拉氏丝虫小鼠模型中。 这项建议的目的是优化恶二唑先驱抑制剂的不间断核糖体拯救 一类新型广谱抗淋病奈瑟菌药物 将该化合物推向研究新药(IND)--实现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。 目的：在目标1中，我们将通过合成孔径雷达驱动的模拟生成来优化导线系列。在《目标2》中我们将 通过体外生物学和ADME评估确定铅系列类似物的优先顺序。在目标3中，我们将确认和 进一步探讨恶二唑类药物不间断拯救核糖体的作用机制及特异性。 在目标4中，我们将根据体内特性选择临床前候选和后备方案。在《目标5》中，我们将 进行IND药代动力学、毒理学和安全性药理学研究。在目标6中，我们将表演 CMC研究，针对一家GMP制造商的1个非GMP药物贴片。在Aim 7中，我们将要求 IND前与FDA的会议。

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.