Delivery of passive immunity against HIV using commensal vaginal Lactobacillus bioengineered to secrete broadly neutralizing domain antibodies

使用经生物工程改造可分泌广泛中和域抗体的共生阴道乳杆菌提供针对 HIV 的被动免疫

• 批准号: 8658658
• 负责人: Laurel A. Lagenaur
• 金额: $ 133.55万
• 依托单位: OSEL, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8658658
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Acute; Address; Antibiotic susceptibility; Antibodies; Ascaridil; Biochemical; Biological; Biological Assay; Biological Response Modifier Therapy; Biomedical Engineering; Cervical; Chromosomal Insertion; Clinical; Codon Nucleotides; Development; Drug Formulations; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epidemiologic Studies; Epitopes; Female; Female of child bearing age; Freeze Drying; Future; Genetic; Genomics; Goals; HIV; HIV Infections; HIV-1; Heterosexuals; Human; Infection; Intravaginal Administration; Investigational Drugs; Lactobacillus; Life; Macaca; Macaca mulatta; Manufacturer Name; Measures; Modeling; Mucous Membrane; Mus; Oryctolagus cuniculus; Passive Immunity; Pharmacologic Substance; Phase; Principal Investigator; Process; Production; Property; Protective Agents; Proteins; Protocols documentation; Recombinants; Research Design; Resistance profile; Risk; Sexually Transmitted Diseases; Site; Small Business Innovation Research Grant; Surface; Tablets; Technology; Testing; Toxic effect; Vagina; Viral; Virus; Virus Diseases; Woman; cost effective; efficacy testing; inhibitor/antagonist; irritation; manufacturing process development; method development; microbicide; nonhuman primate; novel; pathogen; phase 1 study; phase 2 study; pre-clinical; preclinical efficacy; preclinical safety; prevent; programs; promoter; public health relevance; rectal; safety study; safety testing; scale up; self-renewal; simian human immunodeficiency virus; social; transmission process; vaginal lactobacilli

DESCRIPTION: Women bear the brunt of the global AIDS epidemic due in part to their inherent social and biological vulnerability. Biologically, the cervical and vaginal mucosae are vulnerable targets for HIV transmission. In healthy women of childbearing age, mucosal antibodies and microbiota, including Lactobacillus jensenii, help protect these surfaces and serve to block the colonization and infection by intruding pathogens. Epidemiological studies suggest that the loss of vaginal lactobacilli is associated with an increased risk of heterosexual HIV-1 transmission and other sexually transmitted infections. Osel, Inc. has used the properties of these natural protective agents by engineering a native human vaginal Lactobacillus , L. jensenii 1153, to express potent and broadly neutralizing single domain antibodies directed against HIV, termed MucoCept-Ab. Osel recently showed proof-of-concept for this recombinant Lactobacillus approach, by expressing an HIV-1 entry inhibitor mCV-N, from L. jensenii, which reduced SHIV virus transmission in macaques by 63% (Lagenaur et al., Mucosal Immunol, (Jul 6, 2011)). Phase II studies will advance MucoCept-Ab through preclinical development. These studies will focus on developing a safe, efficacious, and stable MucoCept-Ab product, and will include strain optimization, formulation, process development, manufacturing, preclinical safety testing, and efficacy testing in a nonhuman primate (NHP) viral challenge model. The goal of this Phase II is to develop a Lactobacillus strain capable of delivering an antibody locally to prevent HIV infection at the vaginal or rectal mucosa. This strain will be formulated into a cost-effective product capable of persistent protection that is coitally-independent and controlled by women. Aim 1: MucoCept-Ab API optimization and characterization: A) Complete MucoCept-Ab strain optimization: maximize promoter strength, optimize codons for L. jensenii, remove epitope tag, and test two genomic integration sites; B) Complete microbiological characterization of the MucoCept-Ab strain; genetic stability, sequence chromosomal insertion site of expression cassette, biochemical profiling, antibiotic susceptibility/resistant profiling; AP-m36.4 expressio levels; C) Establish ability of the MucoCept-Ab strain to vaginally colonize female rhesus macaques. Aim 2: MucoCept-Ab method development: A) Develop antibodies and ELISA to detect and measure AP-m36.4 levels; B) Develop qPCR assay to measure levels of the MucoCept-Ab strain. Aim 3: MucoCept-Ab formulation, process development, and manufacturing: A) Develop formulation and lyophilization process for production of a stable MucoCept-Ab product; B) Transfer MucoCept-Ab strain and technology to GMP manufacturer for production of MCB and WCB, scale up process development, and production of MucoCept-Ab for preclinical safety studies. Aim 4: MucoCept-Ab preclinical safety and efficacy: A) Evaluate acute toxicity and systemic exposure of the MucoCept-Ab strain and purified AP-m36.4 protein following vaginal administration in mice; B) Evaluate acute toxicity of formulated MucoCept-Ab in the rabbit vaginal irritation model; C) Evaluate subchronic toxicity, vaginal colonization, and AP-m36.4 levels following vaginal administration of the MucoCept-Ab product in female rhesus macaques; D) Evaluate efficacy of the MucoCept-Ab product in female rhesus macaques using a repeated vaginal SHIV challenge protocol. If successful, this approach could provide a safe, yet inexpensive means to deliver passive immunity against HIV to the vaginal mucosa and to address the urgent need for female-controlled approaches to prevent sexually transmitted viral infection.

描述：妇女首当其冲，部分原因是她们固有的社会和生物学脆弱性。从生物学上讲，宫颈粘膜和阴道粘膜是艾滋病毒传播的脆弱靶标。在健康年龄的健康女性中，包括乳酸乳杆菌的粘膜抗体和微生物群，有助于保护这些表面，并通过侵入病原体来阻止定殖和感染。流行病学研究表明，阴道乳酸杆菌的丧失与异性恋HIV-1传播和其他性传播感染的风险增加有关。 Osel，Inc。通过工程设计了人类阴道乳酸杆菌（L. jensenii 1153）来表达针对HIV，称为Mucocept-ab的有效且广泛中和的单个结构域抗体，以表达有效且广泛中和的单域抗体。 OSEL最近通过表达L. Jensenii的HIV-1进入抑制剂MCV-N，显示了这种重组乳杆菌方法的概念验证，该方法将猕猴中的SHIV病毒传播降低了63％（Lagenaur等人，Lagenaur等人，Mucosal Immunol，（Jul 6，2011））。第二阶段的研究将通过临床前开发提高粘液copt-ab。这些研究将着重于开发安全，有效且稳定的粘膜AB产品，并包括在非人类灵长类动物（NHP）病毒挑战模型中进行应变优化，配方，过程开发，制造，临床前安全测试和功效测试。该阶段II的目的是开发能够在局部输送抗体以防止在阴道或直肠粘膜下感染HIV的乳杆菌菌株。该菌株将被配合为具有持久保护的具有成本效益的产品，该产品与女性无关并控制。 AIM 1：粘膜AB API优化和表征：a）完整的Mucocept-AB菌株优化：最大化启动子强度，优化Jensenii的密码子，删除表位标签并测试两个基因组整合位点； b）粘膜AB菌株的完全微生物特征；遗传稳定性，表达盒的序列染色体插入位点，生化分析，抗生素易感性/抗性分析； AP-M36.4表达水平； c）建立粘膜ab菌株的能力，使雌性恒河猕猴在阴道上定位。目标2：粘液Cocecept-Ab方法开发：a）开发抗体和ELISA以检测和测量AP-M36.4级别； b）开发QPCR分析以测量粘膜AB菌株的水平。 AIM 3：粘液Cocecept-ab的配方，过程开发和制造：a）制定制剂和冻干过程，以生产稳定的粘液coctect-ab产品； b）将Mucocept-AB菌株和技术转移给GMP制造商，以生产MCB和WCB，扩展过程开发以及用于临床前安全研究的Mucocept-AB的生产。 AIM 4：粘膜AB临床前的安全性和功效：a）评估小鼠阴道给药后，粘膜cocept-ab菌株和纯化的AP-M36.4蛋白质的急性毒性和全身暴露； b）评估在兔阴道刺激模型中配方粘膜AB的急性毒性； c）评估雌性恒河猕猴的粘液coptagical产物后，评估了阴道施用粘液粘膜产物后的亚慢性毒性，阴道定植和AP-M36.4水平； d）使用重复的阴道SHIV挑战方案评估粘液腹猕猴在雌性猕猴中的功效。 如果成功的话，这种方法可以提供一种安全但廉价的方法，以向阴道粘膜提供对艾滋病毒的被动免疫，并满足对女性控制方法的迫切需求，以防止性传播性传播病毒感染。