Development of Efficacious and Stable Nasal Vaccine Formulations

描述（由申请人提供）：目前，唯一可用于全世界人类使用的佐剂是明矾，这是一种诱导粘膜免疫能力有限的不良佐剂。正在研究另外的佐剂，例如Toll样受体（TLR）配体，但这些佐剂在功效和成本方面是有限的。最近，我们描述了一类新的有效粘膜佐剂，其具有针对广泛疫苗抗原的广泛适用性（Nature Medicine April 20，2008）。我们报道了无针鼻腔给药含有小分子肥大细胞（MC）活化化合物的疫苗制剂，诱发小鼠血清中高水平的抗原特异性IgG和沿着粘膜表面的伊加。这些反应在致死性攻击模型中具有保护性，而且不会产生任何有害的副作用。这些MC活化化合物中的许多是长度约14个氨基酸的小肽序列（肥大细胞活化肽，MCAP），其可以容易且经济地以高纯度制备。MCAP与TLR配体MPL或CpG组合提供优于单独使用的任何佐剂的上级佐剂活性。我们还确定，疫苗抗原的干粉制剂在室温储存后保持效力，并且在鼻免疫后高度有效，这表明它们在运输期间不需要冷藏，同时保持诱导保护性免疫的能力。因此，本项目的目标是开发一种高效、安全的干粉疫苗制剂，用于西尼罗病毒的鼻腔免疫。具体目标是：1。利用重复肽设计确定MC活化肽（MCAP）的最佳结构，以在小鼠中对西尼罗病毒抗原具有最大佐剂活性。2.确定与TLR配体佐剂组合的MC活化肽是否提供能够减少诱导保护性WNV免疫所需的抗原剂量的附加或协同佐剂活性。3.在兔子中开发和测试WNV干粉鼻疫苗制剂，其与人类鼻腔通道具有相同的结构特征。

DESCRIPTION (provided by applicant): Currently, the only adjuvant available for world-wide human use is alum, a poor adjuvant with limited capacity to induce mucosal immunity. Additional adjuvants such as Toll-like receptor (TLR) ligands are being investigated, but these adjuvants are limiting with respect to efficacy and cost. Recently, we described a novel class of potent mucosal adjuvants with broad applicability against a wide range of vaccine antigens (Nature Medicine April 20, 2008). We reported that needle free, nasal administration of vaccine formulations containing small molecule mast cell (MC) activating compounds, evoked high levels of antigen-specific IgG in the serum and IgA along mucosal surfaces in mice. These responses were protective in lethal challenge models, and moreover, did not incur any harmful side effects. Many of these MC activating compounds are small peptide sequences (mast cell activating peptides, MCAP) ~ 14 amino acids in length, which can be readily and economically prepared with high purity. MCAP combined with TLR ligands MPL or CpG provided adjuvant activity that was superior to any adjuvant used alone. We have also determined that dry powder formulations of vaccine antigens maintain potency after room temperature storage and are highly efficacious after nasal immunization, suggesting that they will not require refrigeration during transport while maintaining the ability to induce protective immunity. Therefore the goal of this project is to develop a highly efficacious and safe dry powder vaccine formulation for nasal immunization against West Nile Virus. The Specific Aims are therefore: 1. Utilize reiterative peptide design to determine the optimal structure of MC activating peptides (MCAP) for maximal adjuvant activity with West Nile Virus antigens in mice. 2. Determine if MC activating peptides combined with TLR ligand adjuvants provides additive or synergistic adjuvant activity able to reduce the antigen dose required for induction of protective WNV immunity. 3. Develop and test WNV dry powder nasal vaccine formulations in rabbits, which share structural features with human nasal passages.