Gonorrhea and HIV prevention with intravaginal ring drug delivery

摘要 性传播感染，如艾滋病毒和淋病，是全世界人类健康的主要威胁。 抗生素抗性N.淋病已经在世界各地出现。安全有效的疫苗， 淋病和艾滋病毒仍然难以捉摸。此外，淋病可以增加艾滋病毒的传播约5倍。因此，在本发明中， 迫切需要开发新的方法来预防这些STI。 我们已经鉴定了一种称为2C 7的鼠单克隆抗体（mAb），其针对淋球菌感染。 约95%的临床分离株表达脂寡糖（LOS）表位，具有补体依赖性 杀菌和调理活性并减弱小鼠模型中的淋球菌感染。的嵌合形式 与Genumab持续合作创建的mAb 2C 7（Ximab 2C 7）在以下情况下也有效： 阴道内给予小鼠。在人IgG 1 Fc中引入增加IgG的突变 六聚化和增强补体活化进一步增强Ximab 2C 7的活性。这 麻省大学，行星生物技术公司，橡树峰科学研究所 MassBiologics寻求开发阴道环（IVRs），以联合递送完全人源化的mAb 2C 7 与替诺福韦酯富马酸盐（阴道内TDF;已证明有效预防HIV传播， 妇女），以防止淋病和艾滋病毒感染。 在本提案的R61阶段（第1年和第2年），我们将i）完全人源化mAb 2C 7，其含有 补体增强Fc突变（Humab 2C 7），ii）以相对低的成本产生功能性Humab 2C 7， iii）将Humab 2C 7沿着替诺福韦一起配制成用于局部阴道递送的IVR，和iv） 准备并参加与FDA的IND前会议。 在R33阶段（第3-5年），我们将i）在替诺福韦和Humab给药后进行PK、释放和毒性研究 ii）在绵羊模型中通过IVR递送2C 7， 新型人因子H（FH）和C4 b结合蛋白（C4 BP）双转基因中的多药耐药分离株 小鼠（淋球菌以人类特异性方式结合这些补体抑制剂，因此这些小鼠将提供 Humab 2C 7在人类中必须克服的“障碍”），iii）阐明Humab 2C 7的作用机制， 使用缺乏补体成分（C1 q、C3、C5、C5 aR）和/或吞噬细胞的小鼠进行的Humab 2C 7体内试验 (PMNs巨噬细胞）和iv）开发可扩展的纯化过程和一组药物产品释放 适用于向合同生产组织进行技术转让的检测。 成功完成拟议的工作将产生预防艾滋病毒的临床有效交付成果， 在资助期结束时感染淋病。

ABSTRACT Sexually transmitted infections, such as HIV and gonorrhea, are a major threat to human health worldwide. Antibiotic-resistant N. gonorrhoeae have emerged in all parts of the world. Safe and effective vaccines against gonorrhea and HIV remain elusive. Further, gonorrhea can increase transmission of HIV about 5-fold. Thus, there is an urgent need to develop novel methods to prevent these STIs. We have identified a murine monoclonal antibody (mAb) called 2C7 that is directed against a gonococcal lipooligosaccharide (LOS) epitope that is expressed by ~95% of clinical isolates, has complement-dependent bactericidal and opsonic activity and attenuates gonococcal infection in the mouse model. A chimeric version of mAb 2C7 (Ximab 2C7), created in an ongoing collaboration with Genmab, was also efficacious when administered intravaginally to mice. Introduction of a mutation in human IgG1 Fc that increases IgG hexamerization and enhances complement activation further enhanced activity of Ximab 2C7. This collaborative proposal between UMass, Planet Biotechnology, Inc., Oak Crest Institute of Science and MassBiologics seeks to develop intravaginal rings (IVRs) to deliver fully humanized mAb 2C7 in combination with tenofovir disoproxil fumarate (intravaginal TDF; proven efficacious in preventing HIV transmission to women) to prevent gonorrhea and HIV infections. In the R61 phase (years 1 and 2) of this proposal, we will i) fully humanize mAb 2C7 that contains the complement-enhancing Fc mutation (Humab 2C7), ii) produce functional Humab 2C7 at a relatively low cost in tobacco plants, iii) formulate Humab 2C7 along with the tenofovir into IVRs for local vaginal delivery and iv) prepare for and participate in a pre-IND meeting with the FDA. In the R33 phase (years 3-5) we will i) perform PK, release and toxicity studies following tenofovir and Humab 2C7 delivery via IVR in an ovine model, ii) test the efficacy of Humab 2C7 against contemporary clinical multidrug-resistant isolates in novel human factor H (FH) and C4b-binding protein (C4BP) dual transgenic mice (gonococci bind these complement inhibitors in a human specific manner, thus these mice will provide the `obstacles' that Humab 2C7 will have to surmount in humans), iii) elucidate the mechanism of action of Humab 2C7 in vivo using mice that lack complement components (C1q, C3, C5, C5aR) and /or phagocytes (PMNs, macrophages) and iv) develop a scalable purification process and a set of drug product release assays appropriate for technology transfer to a contract manufacturing organization. Successful completion of the proposed work will result in a clinically effective deliverable to prevent HIV and gonorrhea at the end of the funding period.