Novel VLP Adjuvant based on the VesiVax system

描述(申请人提供)：分子快递公司专门从事用于药物输送和疫苗开发的功能化脂质体的设计和工程。在以前的研究中，人们筛选了一系列广泛的脂质体配方，以建立满足某些物理化学参数并在动物身上提供强大免疫反应的最佳脂类组合。这些单层囊泡的直径约为100 nm，在生物缓冲溶液中稳定(即一年以上没有聚集、沉淀或成分降解)，并且可商业扩展。免疫原脂质体组合物构成了称为VesiVax(R)系统的疫苗平台技术的基础。VesiVax(R)系统被设计成易于操作，因此目标抗原可以从免疫原脂质体的表面展示出来，供随后的免疫研究使用。基于VesiVax(R)系统的疫苗可以使用两种形式进行工程设计：i)靶抗原或佐剂蛋白可以被设计为具有疏水结构域(HD)的融合蛋白，这有助于将其整合到脂质体中；或者ii)靶抗原或佐剂以蛋白质、多肽或碳水化合物的形式通过接合连接到脂质体表面(即可结合佐剂脂囊泡；CALV)。VesiVax(R)平台的灵活性允许将抗原和佐剂一起制成脂质体，从而使免疫反应最大化。在这份SBIR AT第一阶段提案中，我们打算建立VesiVax(R)CALV平台作为病毒样颗粒(VLP)的有效佐剂的效用。为了证明这一概念，我们建议制备包含不同Toll样受体(TLR)激动剂的VesiVax(R)Calv制剂，然后将它们与设计用于表达HIV抗原的VLP(HIV-VLP)结合(特定目标1)。VesiVax(R)Calv HIV-VLP随后将在小鼠和兔模型上进行测试(特定目标2)。这些研究可能具有很高的影响力，因为这些研究表明：i)VesiVax(R)CALV可以佐剂VLP将通过增加CALV系列研究试剂盒的销售产生立竿见影的影响；ii)舌下给药途径刺激强大的免疫反应将是VesiVax(R)技术的另一个有用的应用；3)一种或多种VesiVax(R)Calv HIV-VLP配方可在体外测试中激发强大的HIV抗体中和活性，这将为挑选其中一种或多种候选者在非人类灵长类动物中进行进一步测试奠定基础，之后将最终选出VesiVax(R)Calv HIV-VLP候选者，并将其推向临床测试。这种疫苗将为可在全球范围内实施的具有成本效益的艾滋病毒免疫战略提供基础。

DESCRIPTION (provided by applicant): Molecular Express specializes in the design and engineering of functionalized liposomes for drug delivery and vaccine development. In previous studies, an extensive set of liposome formulations were screened to establish the optimal lipid combination that meets certain physicochemical parameters and provides strong immune responses in animals. These unilamellar vesicles have diameters of approximately 100 nm, are stable (i.e., no aggregation, precipitation or degradation of the components for over a year) in biological buffer solutions, and are commercially scalable. The immunogenic liposome compositions form the basis of a vaccine platform technology called the VesiVax(r) system. The VesiVax(r) system was designed to be easily manipulated, so that target antigens could be displayed from the surface of the immunogenic liposomes for subsequent use in immunization studies. Vaccines based on the VesiVax(r) system can be engineered using two formats: i) target antigens or adjuvant proteins can be engineered to be expressed as a fusion protein with a hydrophobic domain (HD), which facilitates its incorporation into the liposomes or; ii) target antigens or adjuvants in the form of proteins, peptides or carbohydrates can be attached via conjugation to the surface of the liposomes (i.e., conjugatable adjuvant lipid vesicles; CALVs). The flexibility of the VesiVax(r) platform allows both antigens and adjuvants to be formulated together into the liposomes thus maximizing the immune response. In this SBIR AT Phase I proposal, we intend to establish the utility of the VesiVax(r) CALV platform as an effective adjuvant for virus-like particles (VLPs). To demonstrate this concept, we propose to prepare VesiVax(r) CALV formulations containing different Toll-like Receptor (TLR) agonists and then conjugating them to VLPs that have been designed to express HIV antigens (HIV-VLPs) (Specific Aim 1). The VesiVax(r) CALV HIV-VLPs will then be tested in mouse and rabbit models (Specific Aim 2). These studies are potentially of high impact because demonstration that: I) the VesiVax(r) CALVs can adjuvant VLPs would have an immediate impact through increasing sales of the CALV line of research kits; II) the sublingual route of administration stimulates potent immune responses would be another useful application of the VesiVax(r) technology and; III) one or more VesiVax(r) CALV HIV-VLP formulations stimulate potent HIV antibody neutralization activity in the in vitro assays will set the stage for the selection of one or more of these candidates to undergo further testing in non-human primates prior to final selection of a VesiVax(r) CALV HIV-VLP candidate that will be advanced to clinical testing. Such a vaccine will provide the basis for a cost effective HIV immunization strategy that could be implemented on a global scale.