Novel VLP Adjuvant based on the VesiVax system

描述（由申请人提供）：Molecular Express专门从事用于药物递送和疫苗开发的功能化脂质体的设计和工程。在先前的研究中，筛选了一组广泛的脂质体制剂，以建立满足某些理化参数并在动物中提供强免疫应答的最佳脂质组合。这些单层囊泡具有约100 nm的直径，是稳定的（即，在生物缓冲溶液中一年以上没有组分的聚集、沉淀或降解），并且是商业上可放大的。免疫原性脂质体组合物形成称为VesiVax系统的疫苗平台技术的基础。VesiVax（r）系统设计为易于操作，使得靶抗原可以从免疫原性脂质体的表面展示，用于随后的免疫研究。基于VesiVax系统的疫苗可以使用两种形式进行工程化：i）靶抗原或佐剂蛋白可以被工程化以表达为具有疏水结构域（HD）的融合蛋白，这有助于其掺入脂质体中;或ii）蛋白质、肽或碳水化合物形式的靶抗原或佐剂可以通过缀合附接至脂质体的表面（即，可缀合的佐剂脂质囊泡; CALV）。VesiVax（r）平台的灵活性允许抗原和佐剂一起配制成脂质体，从而最大限度地提高免疫应答。 在SBIR AT I期提案中，我们打算将VesiVax（r）CALV平台作为病毒样颗粒（VLP）的有效佐剂。为了证明这一概念，我们建议制备含有不同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）偶联（具体目标1）。然后将在小鼠和家兔模型中测试VesiVax（r）CALV HIV-VLP（特定目标2）。这些研究具有潜在的高影响力，因为证明：I）VesiVax（r）CALV可以辅助VLP，将通过增加CALV系列研究试剂盒的销售产生直接影响; II）舌下给药途径刺激有效的免疫应答将是VesiVax（r）技术的另一个有用应用; III）一种或多种VesiVax（r）CALV HIV-VLP制剂在体外试验中刺激有效的HIV抗体中和活性，这将为选择一种或多种这些候选药物进行进一步的非药物试验奠定基础。在最终选择将进入临床试验的VesiVax（r）CALV HIV-VLP候选物之前，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奠定基础。

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.