Ll capsomeres as a next generation preventive HPV vaccine

Ll 壳粒作为下一代预防性 HPV 疫苗

• 批准号: 7727548
• 负责人: Robert L Garcea
• 金额: $ 31.33万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7727548
• 关键词: Adjuvant; Adverse effects; Animal Experiments; Antibodies; Antigens; Award; Canis familiaris; Capsid; Cell Culture Techniques; Cells; Chimeric Proteins; Clinical; Clinical Trials; Cold Chains; Country; Cytotoxic T-Lymphocytes; Development; Dose; Drug Formulations; Escherichia coli; Gardasil; Generations; Genital Human Papilloma Virus Infection; Goals; HPV-High Risk; Human; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papillomavirus 16; Humoral Immunities; Immune; Immune response; Immune system; Immunity; Immunization; Immunology; Infection; Inferior; Instruction; Knowledge; L1 viral capsid protein; Laboratory Study; Life; Link; Malignant neoplasm of cervix uteri; Medicine; Modeling; Mus; Patients; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Placebos; Preparation; Preventive; Production; Property; Proteins; Receptor Activation; Recombinants; Refrigeration; Reproduction spores; Research Personnel; Scheme; Specimen; Structure; Temperature; Testing; Time; Toll-like receptors; Toxic effect; Toxicology; Vaccinated; Vaccination; Vaccines; Vagina; Virus-like particle; Woman; Women' s Health; aluminum sulfate; base; cost; crosslink; immunogenic; immunogenicity; mouse model; neutralizing antibody; next generation; preclinical study; prevent; programs; prophylactic; recombinant virus; response; success

The recent development of prophylactic vaccines protective against "high risk" human papillomaviruses (HPV) is a landmark in medicine. The current vaccine is composed of recombinant virus-like particles (VLPs) of the major capsid protein, L1, including the high risk HPV types 16 and 18. Phase ll/lll clinical trials have shown >95% efficacy of these VLP preparations in preventing HPV 16, 18 infection, and thus hopefully preventing the eventual development of about 70% of cervical cancers associated with these types. Despite this success the VLP vaccines have the adverse attributes of high cost ($360 for the initial three immunizations In the USA) and a requirement for refrigeration {i.e., cold chain). We have previously characterized the immunogenic properties of VLP subunits, pentameric L1 capsomeres, and compared these "subunits" to VLPs in a canine infection model. In this model capsomeres appeared near equivalent to VLPs in inducing protection against infection. Capsomeres are purified after recombinant expression of L1 in E. coli as a GST-fusion protein, at levels suggesting a significant reduction in manufacturing expense. The protein is readily purified, and can be precipitated, resuspended, and stored at room temperature without loss of immunogenicity. In the context of an ongoing NCI RAPID award GST-L1 capsomeres have been proposed as a "next generation" HPV vaccine that might be ideally suited for production and use in underdeveloped countries of the world where cervical cancer is particularly prevalent. We propose to use the GST-L1 protein that has been GMP produced (by Shantha Biotechnica), vialed, and toxicology screened under the auspices of a RAPID award for study in a phase I human trial. In aims 1 and 2 we propose a dose escalation scheme of 15 subjects at each of three dose levels (10, 50, 500 micrograms and placebo), repeated three times, with concomitant analysis of toxicity, development of neutralizing antibodies, cytotoxic T-cell responses, and possible adverse effects of GST antibody development. In the third aim, laboratory studies with clinical specimens and animal experiments will test new adjuvant strategies including Toll-like receptor activation, and evaluate the effects on humoral immunity of antigen spacing and linking by comparing capsomeres, cross-linked capsomeres, and VLPs. Knowledge of L1 capsomere structure and interaction with the innate immune system will be utilized to generate longer-lived and broader immunity with lower and fewer doses. The results of this project will determine whether the GST-LI preparation can proceed to further phase II testing. RELEVANCE (See instructions): Persistent infection with high risk HPV is a necessary cause of cervical cancer. Our goal is the development of a next generation, non-inferior HPV preventive vaccine that is economically feasible for production for use in the underdeveloped world where the impact of cervical cancer on women's health is greatest.

针对“高危”人乳头瘤病毒的预防性疫苗的最新发展 (HPV)是医学上的里程碑目前的疫苗由重组病毒样颗粒组成 主要衣壳蛋白L1的VLP，包括高危型HPV 16和18。II/III期临床试验 已经显示这些VLP制剂在预防HPV 16、18感染方面的功效>95%，因此有希望 预防与这些类型相关的约70%的宫颈癌的最终发展。尽管 这种成功的VLP疫苗具有高成本的不利属性（最初的三种疫苗为360美元 在美国的免疫接种）和对冷藏的要求（即，冷链）。我们先前已经 表征了VLP亚基，五聚体L1壳粒的免疫原性，并比较了 这些“亚基”在犬感染模型中转化为VLP。在这个模型中，壳粒似乎几乎相当于 VLP诱导抗感染保护。在L1的重组表达后纯化壳粒 在大肠大肠杆菌作为GST-融合蛋白，在水平上表明在制造费用的显着降低。 蛋白质易于纯化，并且可以沉淀、重悬和在室温下储存 不损失免疫原性。在正在进行的NCI RAPID奖的背景下，GST-L1壳粒具有 被提议作为“下一代”HPV疫苗，可能非常适合在 子宫颈癌是宫颈癌的主要病因之一。我们建议使用 GST-L1蛋白已通过GMP生产（由Shantha Biotechnica生产）、装瓶和毒理学筛选 在第一阶段人体试验研究的RAPID奖的赞助下。在目标1和2中，我们提出了一种剂量， 三个剂量水平（10、50、500微克和安慰剂）各15例受试者的递增方案， 重复三次，同时分析毒性、中和抗体的产生、细胞毒性 T细胞反应和GST抗体产生的可能不良影响。第三，实验室 临床标本和动物实验研究将测试新的佐剂策略，包括Toll样 受体激活，并评估抗原间隔和连接对体液免疫的影响， 比较壳粒、交联壳粒和VLP。了解L1壳粒结构， 与先天免疫系统的相互作用将被用来产生更长寿命和更广泛的免疫力， 更低更少的剂量。该项目的结果将决定GST-LI制剂是否能够 继续进行进一步的第二阶段测试。 相关性（参见说明）： 高危型HPV的持续感染是宫颈癌的一个必要原因。我们的目标是发展 下一代非劣效HPV预防性疫苗的生产在经济上是可行的 在不发达国家，宫颈癌对妇女健康的影响最大。