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，筛选了大量脂质体配方，以建立符合某些物理化学参数的最佳脂质组合，并在动物中提供强大的免疫反应。这些单层囊泡直径约为100纳米，在生物缓冲溶液中稳定（即一年以上不会聚集、沉淀或降解成分），并且具有商业可扩展性。免疫原性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)系统设计为易于操作，因此可以从免疫原性脂质体表面显示目标抗原，以便随后用于免疫研究。基于VesiVax(r)系统的疫苗可以采用两种形式进行工程设计：i)可以设计靶抗原或佐剂蛋白，使其表达为具有疏水结构域（HD）的融合蛋白，从而促进其融入脂质体；ii)蛋白、多肽或碳水化合物形式的靶抗原或佐剂可通过偶联方式附着在脂质体表面（即，可偶联佐剂脂囊；calv）。VesiVax(r)平台的灵活性允许抗原和佐剂一起配制到脂质体中，从而最大限度地提高免疫反应。在这个SBIR AT I期提案中，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，我们建议制备含有不同toll样受体（TLR）激动剂的VesiVax(r) CALV制剂，然后将它们偶联到设计用于表达HIV抗原的VLPs （HIV-VLPs）上（Specific Aim 1）。VesiVax(r) CALV HIV-VLPs随后将在小鼠和兔子模型中进行测试（Specific Aim 2）。这些研究可能具有很高的影响，因为证明：I) VesiVax(r) CALV可以辅助VLPs，将通过增加CALV研究试剂盒系列的销售产生直接影响；II)舌下给药途径刺激有效的免疫反应将是VesiVax(r)技术的另一个有用的应用；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，将为选择一种或多种候选药物在非人灵长类动物中进行进一步测试奠定基础，然后最终选择一种VesiVax(r) CALV HIV- vlp候选药物进行临床测试。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.