Novel immunotherapeutics against multidrug-resistant Neisseria gonorrhoea

针对多重耐药淋病奈瑟菌的新型免疫疗法

• 批准号: 10207360
• 负责人: SANJAY RAM
• 金额: $ 86.28万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-11 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207360
• 关键词: Academia; Amino Acids; Anti-Infective Agents; Antibiotics; Antimicrobial Resistance; Attenuated; Binding; Biological Sciences; Biotechnology; Cell surface; Cells; Centers for Disease Control and Prevention (U.S.); Clinical; Clostridium difficile; Collaborations; Complement; Complement Activation; Complement Factor H; Complement Inactivators; Complement Membrane Attack Complex; Cytolysis; Drug resistance; Drug-resistant Neisseria Gonorrhoeae; Ectopic Pregnancy; Epitopes; Gonorrhea; Head; Health; High Risk Woman; Human; Immunoglobulin G; Immunotherapeutic agent; Immunotherapy; In Vitro; Individual; Industry; Infection; Infertility; Institutes; Lead; Length; Macaca fascicularis; Mediating; Microbe; Modeling; Monoclonal Antibodies; Multi-Drug Resistance; Mus; Mutation; N-terminal; Neisseria gonorrhoeae; Nicotiana; Pathogenesis; Pelvic Inflammatory Disease; Pharmaceutical Preparations; Planets; Plants; Point Mutation; Polysaccharides; Positioning Attribute; Preparation; Production; Proteins; Rattus; Reporting; Resistance; Sexually Transmitted Agents; Sexually Transmitted Diseases; Superbug; Surface; System; Tail; Testing; Tissues; Tobacco; Topical application; Toxic effect; Toxicokinetics; Toxicology; Transgenic Mice; United States; Vagina; Vaginal Ring; Virulence; Woman; arm; bactericide; carbapenem-resistant Enterobacteriaceae; comparative; complement 4b-binding protein; cost; cross reactivity; drug development; human monoclonal antibodies; immunogenic; immunoprophylaxis; in vivo; lipooligosaccharide; macrophage; man; meetings; men; multi-drug resistant pathogen; novel; novel therapeutics; pathogen; pre-clinical; preclinical development; synergism; vaccine candidate

ABSTRACT In 2013, the CDC listed Neisseria gonorrhoeae (Ng) is listed as one of the three multidrug-resistant pathogens that represented ‘urgent’ threats to human health worldwide. Ng has become resistant to almost every antibi-otic and has achieved “superbug” status. Complications of gonorrhea include pelvic inflammatory disease, which may lead to infertility and ectopic pregnancy. Novel therapeutics against this pathogen are urgently needed. This collaboration between industry and academia seeks to further preclinical development of two immunotherapeutics against drug-resistant Ng. Each molecule targets a distinct epitope on Ng; both are im-portant for pathogenesis and thus ubiquitously expressed in vivo. Advantages of targeting distinct epitopes im-portant for virulence include i) synergistic activity and ii) raising the barrier for the development of drug re-sistance, if it were to occur. A chimeric mAb targets Ng and mediates complement (C′)-dependent killing of Ng in vivo. Several pathogens including Ng evade C′ by binding to a host C′ inhibitor called factor H (FH). FH comprises 20 domains, arranged in an extended head-to-tail fashion. FH domains 18-20 (lacks C′ inhibitory function) fused to IgG Fc effects C′-dependent killing of Ng. FH domains 19 and 20 are important to limit un-wanted C′ activation on host tissue. We introduced a D→G amino acid point mutation in domain 19 of FH18-20/Fc. This protein, called FH*/Fc, retains its efficacy against Ng in vitro and in vivo but does not lyse host cells. In Aim 1 Planet Biotechnology, Inc. will produce the chimeric mAb and FH*/Fc in tobacco plants. This will permit efficient and low cost production. The efficacy of plant-produced FH*/Fc and the chimeric mAb against Ng in vitro and in mice will be tested by UMass in Aim 2. The drugs will be administered: i) systemically (mod-eling adjunctive treatment of established infection in men and women) or ii) intravaginally (modeling topical immunoprophylaxis in high-risk women). Each molecule will be tested individually and in combination for syn-ergy and to raise the barrier for drug resistance. Efficacy will also be assessed in novel transgenic (Tg) mice that express the human C′ inhibitors, FH and C4b-binding protein, which Ng bind to in a human-specific man-ner to evade C′, and therefore represent barriers to the drugs that may be encountered in humans. In Aim 3, Oak Crest Institute will formulate both drugs in combination in slow-release vaginal rings for use as topical im-munoprophylactics. In Aim 4 Xenometrics will perform toxicology and toxicokinetic (TK) studies in rats and cynomolgus monkeys and Comparative Biosciences will perform tissue-cross reactivity studies. In Aim 5, the mechanism of action of the chimeric mAb and FH*/Fc and will be defined in vivo using mice that lack C3, C5, PMNs and/or macrophages to define correlates of protection in vivo. These studies may lead to preparation for and participation in a pre-IND meeting with FDA (Aim 6).

摘要 2013年，美国疾病控制和预防中心将淋病奈瑟菌（Ng）列为三种多重耐药病原体之一，对全球人类健康构成“紧迫”威胁。Ng已经对几乎所有抗生素产生耐药性，并获得了“超级细菌”的地位。淋病的并发症包括盆腔炎，这可能导致不孕和异位妊娠。迫切需要针对这种病原体的新疗法。工业界和学术界之间的这种合作旨在进一步临床前开发两种针对耐药Ng的免疫治疗药物。每个分子靶向Ng上的不同表位;两者对于发病机制都很重要，因此在体内普遍表达。靶向对毒力重要的不同表位的优势包括i）协同活性和ii）提高耐药性发展的屏障（如果发生的话）。嵌合mAb靶向Ng并介导体内Ng的补体（C′）依赖性杀伤。包括Ng在内的几种病原体通过与称为因子H（FH）的宿主C′抑制剂结合来逃避C′。FH包含20个结构域，以扩展的头对尾方式排列。与IgG Fc融合的FH结构域18-20（缺乏C′抑制功能）影响Ng的C′依赖性杀伤。FH结构域19和20对于限制宿主组织上不需要的C′活化是重要的。我们在FH 18 -20/Fc的结构域19中引入了一个D→G的氨基酸点突变。这种蛋白质称为FH*/Fc，在体外和体内保留其对Ng的功效，但不裂解宿主细胞。In Aim 1 Planet Biotechnology，Inc.将在烟草植物中产生嵌合mAb和FH*/Fc。这将允许有效和低成本的生产。植物产生的FH*/Fc和嵌合mAb在体外和小鼠中抗Ng的功效将由UMass在Aim 2中测试。这些药物将通过以下方式给药：i）全身给药（对男性和女性中已确定感染进行模式化预防性治疗）或ii）阴道内给药（对高危女性进行模式化局部免疫预防）。每种分子都将单独或组合进行测试，以提高耐药性的屏障。还将在表达人C′抑制剂FH和C4 b结合蛋白的新型转基因（Tg）小鼠中评估疗效，Ng以人类特异性方式与其结合以逃避C′，因此代表了人类可能遇到的药物屏障。在目标3中，Oak Crest研究所将在缓释阴道环中配制两种药物的组合，用作局部免疫抑制剂。在目标4中，Xenometrics将在大鼠和食蟹猴中进行毒理学和毒代动力学（TK）研究，Comparative Biosciences将进行组织交叉反应性研究。在目的5中，嵌合mAb和FH*/Fc的作用机制将使用缺乏C3、C5、PMN和/或巨噬细胞的小鼠在体内定义，以定义体内保护的相关性。这些研究可能导致准备和参与与FDA的IND前会议（目标6）。

项目成果

Antibacterial Fusion Proteins Enhance Moraxella catarrhalis Killing.

抗菌融合蛋白增强卡他莫拉菌的杀灭作用。

• DOI: 10.3389/fimmu.2020.02122
• 发表时间: 2020
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Laabei,Maisem;Colineau,Lucie;Bettoni,Serena;Maziarz,Karolina;Ermert,David;Riesbeck,Kristian;Ram,Sanjay;Blom,AnnaM
• 通讯作者: Blom,AnnaM

An optimized Factor H-Fc fusion protein against multidrug-resistant Neisseria gonorrhoeae.

• DOI: 10.3389/fimmu.2022.975676
• 发表时间: 2022
• 期刊: Frontiers in immunology
• 影响因子: 7.3

A Novel Sialylation Site on Neisseria gonorrhoeae Lipooligosaccharide Links Heptose II Lactose Expression with Pathogenicity.

淋病奈瑟菌脂寡糖上的新型唾液酸化位点将庚糖 II 乳糖表达与致病性联系起来。

• DOI: 10.1128/iai.00285-18
• 发表时间: 2018
• 期刊: Infection and immunity
• 影响因子: 3.1
• 作者: Ram,Sanjay;Gulati,Sunita;Lewis,LisaA;Chakraborti,Srinjoy;Zheng,Bo;DeOliveira,RosaneB;Reed,GeorgeW;Cox,AndrewD;Li,Jianjun;StMichael,Frank;Stupak,Jacek;Su,Xiao-Hong;Saha,Sudeshna;Landig,CorinnaS;Varki,Ajit;Rice,PeterA
• 通讯作者: Rice,PeterA

Development of Complement Factor H-Based Immunotherapeutic Molecules in Tobacco Plants Against Multidrug-Resistant Neisseria gonorrhoeae.

• DOI: 10.3389/fimmu.2020.583305
• 发表时间: 2020
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Shaughnessy J;Tran Y;Zheng B;DeOliveira RB;Gulati S;Song WC;Maclean JM;Wycoff KL;Ram S
• 通讯作者: Ram S

Human Factor H Domains 6 and 7 Fused to IgG1 Fc Are Immunotherapeutic against Neisseria gonorrhoeae.

与 IgG1 Fc 融合的人因子 H 结构域 6 和 7 可针对淋病奈瑟菌进行免疫治疗。

• DOI: 10.4049/jimmunol.1701666
• 发表时间: 2018
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Shaughnessy,Jutamas;Lewis,LisaA;Zheng,Bo;Carr,Caleb;Bass,Isaac;Gulati,Sunita;DeOliveira,RosaneB;Gose,Severin;Reed,GeorgeW;Botto,Marina;Rice,PeterA;Ram,Sanjay
• 通讯作者: Ram,Sanjay