Sialic acid analogs against multidrug-resistant gonorrhea

摘要 淋病是全球主要的公共卫生问题。全球约有8700万新淋病病例 每年一次。2018年，美国报告了583,405例病例，自 2009年的历史低点。同时感染艾滋病毒和淋病会增加艾滋病毒的传播率。 收牌。阴道微生物群失调(即细菌性阴道病(BV))增加淋病和 艾滋病毒的获取和传播。女性淋病的严重后遗症包括不孕、异位 怀孕和慢性盆腔疼痛。淋病奈瑟菌(Ng)已对几乎所有抗生素产生抗药性 在临床上使用。几乎每个大陆对头孢曲松和阿奇霉素的耐药性报告预示着一种 无法治愈的淋病时代。开发针对Ng的新疗法是全球公共卫生的优先事项。 淋球菌采用一种独特的免疫逃避策略，将其脂寡糖(LOS)的上限 使用宿主来源的CMP-Neu5Ac的表面唾液酸转移酶(LST)。LOS分析使Ng得以逃避 宿主免疫的几个方面，包括补体和阳离子抗菌肽(CAMP)。我们 发现在淋球菌中添加了非磺酸盐类(CMP-Nulos)，如MP-军团酸类(CMP-Nulos)。 Leg5，7Ac2)和CMP-酮基脱氧农磺酸盐(CMP-KDN)用这些Nulo来封顶它们的LO。成立为法团 进入Ng LOS的Nulo使细菌容易受到补体和cAMP的影响。重要的是，CMP-Nulos 显著缩短Ng在小鼠阴道内定植的时间和负担。放线菌素(该委员会成员 在《消除对妇女一切形式歧视公约》的《行动纲领》中发挥了关键作用。我们的有效性、安全性和稳定性研究 建立CMP-Leg5，7Ac2和CMP-KDn作为阴道分娩预防Ng的有前景的候选方案 收购。在目标1的第1-2年中，NRC将为本建议书中的所有工作生成CMP-Nulo，执行 药物经济学分析和放大评估。过程开发、质量释放分析、开发 已确定的铅在模拟生殖道液中的效力分析和稳定性将在3-5年进行。目标 2将阐明可能影响CMP-Nulos安全疗效的因素。其中包括CMP零的影响 有利或不利的阴道细菌的生长和活力，唾液酸酶的影响，由BV- 相关病原体和自然产生的抗Nulo抗体对CMP-Nulos疗效的影响 在体内，检测宫颈阴道分泌物中CMP-Nulo水解酶的活性，并测试其特异性 坎普将结合含Nulo的多糖，以完善针对Ng的CMP-Nulo的MOA。目标3将制定 在体外和绵羊模型中进行PK研究，并检查 IVRs上阴道微生物区系生物被膜的形成。在目标1-3的第1-2年中开展的工作将确定一条线索； 第三至第五年的工作将侧重于进一步的产品开发和支持IND的活动。AIM 4将验证 Cpn-Nulo对遗传和地理差异的Ng分离株的活性。目标5包括 对所有目标的监管指导和监督，最终将在IND前与FDA举行会议。

ABSTRACT Gonorrhea is a major public health problem globally. About 87 million new cases of gonorrhea occur worldwide annually. In 2018, 583,405 cases were reported in the U.S, an 82.6% increase in disease incidence since the historic low in 2009. Concomitant infection with HIV and gonorrhea can increase rates of HIV transmission 5- fold. Dysbiosis of the vaginal microbiome (i.e., bacterial vaginosis (BV)) increases the risk of gonorrhea and HIV acquisition and transmission. Serious sequelae of gonorrhea in women include infertility, ectopic pregnancy and chronic pelvic pain. Neisseria gonorrhoeae (Ng) has become resistant to almost every antibiotic in clinical use. Reports of resistance to ceftriaxone and azithromycin from almost every continent portends an era of untreatable gonorrhea. Development of novel therapies against Ng is a global public health priority. Gonococci deploy a unique immune evasion strategy wherein it caps its lipooligosaccharide (LOS) by a surface LOS sialyltransferase (Lst) using host-derived CMP-Neu5Ac. LOS sialylation enables Ng to evade several aspects of host immunity, including complement and cationic antimicrobial peptides (CAMPs). We discovered that gonococci fed with CMP-nonulosonates (CMP-NulOs), such as CMP-legionaminic acid (CMP- Leg5,7Ac2) and CMP-ketodeoxynonulosonate (CMP-Kdn) cap their LOS with these NulOs. Incorporation of NulOs into Ng LOS renders bacteria susceptible to complement and CAMPs. Importantly, CMP-NulOs significantly shorten the duration and burden of Ng vaginal colonization in mice. Cathelicidin (a member of the CAMP family) played a key role in the MOA of CMP-NulOs. Our efficacy, safety and stability studies have established CMP-Leg5,7Ac2 and CMP-Kdn as promising candidates for intravaginal delivery to prevent Ng acquisition. In years 1-2 of Aim 1, the NRC will produce CMP-NulOs for all work in this proposal, perform a pharmacoeconomic analysis and assess scale-up. Process development, quality release assays, development of potency assays and stability in simulated genital tract fluids of the identified lead will follow in years 3-5. Aim 2 will elucidate factors that may affect safety efficacy of CMP-NulOs. These include effects of CMP-NulOs on growth and viability of favorable or unfavorable vaginal bacteria, the effects of sialidases elaborated by BV- associated pathogens and naturally-occurring anti-NulO antibodies on the efficacy of CMP-NulOs in vitro and in vivo, examining cervicovaginal secretions for CMP-NulO hydrolase activity and testing the specificity of CAMPs to bind NulO-containing glycans to refine the MOA of CMP-NulO against Ng. Aim 3 will formulate the CMP-NulOs into intravaginal rings (IVRs), perform PK studies in vitro and in a sheep model, and examine biofilm formation by vaginal microbiota on IVRs. Work in years 1-2 of the Aims 1-3 will identify a single lead; works in years 3-5 will focus on further product development and IND-enabling activities. Aim 4 will verify activity of the lead CMP-NulO against genetically and geographically diverse Ng isolates. Aim 5 includes regulatory guidance and oversight for all Aims that will culminate in a pre-IND meeting with the FDA.