Designer covalent heterobivalent inhibitors to prevent IgE-dependent allergic reactions

项目摘要： 过敏反应是由过敏原介导的免疫球蛋白E（IgE）抗体聚集引起的 在肥大细胞表面，这是启动全身反应的“关键事件”。过敏性疾病 发达国家的健康问题日益严重。根据过敏反应的严重程度， 可以从简单的瘙痒到过敏性休克，在美国每年导致1,500人死亡。那里 没有治愈过敏症的方法，目前的疗法集中在治疗急性症状或慢性免疫性疾病。 镇压即使在最严格的饮食限制下，意外接触食物过敏也是非常频繁的， 有发生危及生命的过敏反应风险的患者。因此，需要更有效的替代治疗 IgE介导的过敏反应。本申请的目的是开发过敏原特异性IgE 抑制剂（cHBI：共价异二价抑制剂），靶向过敏原结合IgE，并防止其 识别过敏原这种方法在特异性靶向开始之前抑制过敏反应， 没有广泛的免疫抑制天然存在的过敏原通常在结构上是复杂的， 具有多个过敏诱导表位的异源蛋白质。从历史上看， 单个过敏原表位对整体过敏反应的贡献。在过去的融资中 循环，我们通过开发多组分表位呈递平台解决了这个问题， 叫做纳米过敏原利用纳米过敏原筛选，我们可以研究单个过敏原的免疫原性 表位，并鉴定免疫显性表位。使用鉴定的免疫显性表位，我们合成了 模拟表位并阻断识别特异性表位的IgE抗体的cHBI抑制剂。这 方法抑制IgE识别过敏原，当它进入系统，并防止启动过敏原， 过敏反应。我们最近证明，使用cHBIs的鸡尾酒，我们可以阻止花生， 在体外细胞脱粒方法中使用患者样品进行过敏反应。在当前应用中， 我们将使用人源化小鼠模型评价花生过敏抑制的体内效果。同时，我们将 利用cHBI作为阻断其他过敏原的平台的潜力，包括贝类，尘螨和 榛子这项工作具有创新性和重要意义，因为它（i）提供了一种新的分子设计 抑制引发过敏反应的“关键事件”的方法，具有潜在的长期临床应用 在食物，环境和药物过敏，（ii）它这样做没有任何非特异性抑制免疫 系统组件，和（iii）开发一个非常需要的，生理相关的，易于调整， 可重复的平台（纳米过敏原），将用于识别免疫显性公共过敏原 表位及其在脱粒反应期间的相对意义。最后，该应用程序具有高 潜在影响，因为cHBI有可能转化为临床。

Project Summary: Allergic reactions are elicited by the allergen-mediated-clustering of the immunoglobulin E (IgE) antibodies on the surface of mast cells, which is “the key event” in initiation of the systemic reaction. Allergic diseases are increasing health concerns in developed nations. Depending on the severity of the allergic reaction, the results can vary from a simple itch to anaphylactic shock, which results in 1,500 deaths each year in the US. There are no cures for allergies, and current therapies focus on treatment of acute symptoms or chronic immune suppression. Even with the most restricted diets, accidental exposure is very frequent for food allergies, putting patients at risk for life threatening anaphylaxis. Thus, there is a need for more effective, alternative treatments for IgE-mediated allergic responses. The objective of this application is to develop allergen-specific IgE inhibitors (cHBI: covalent heterobivalent inhibitor) that target allergen-binding IgE, and prevent it from recognizing the allergen. This approach inhibits the allergic reaction before it starts with specific targeting but without broad immune suppression. Naturally occurring allergens are typically complex, structurally heterogenous proteins with multiple allergy-inducing epitopes. Historically it has been a challenge to identify contributions from individual allergen epitopes to the overall allergic reaction. During the previous funding cycle, we addressed this problem by developing a multicomponent epitope presentation platform that we named nanoallergens. Using nanoallergen screening, we can study the immunogenicity of individual allergen epitopes, and identify immunodominant epitopes. Using the identified immunodominant epitope, we synthesize cHBI inhibitors that mimic the epitope and block the IgE antibodies that recognize the specific epitope. This method inhibits the IgE from recognizing the allergen when it enters the system and prevents the initiation of the allergic reaction. We have recently demonstrated that using a cocktail of cHBIs, we can block peanut allergic reactions using patient samples in an in vitro cellular degranulation method. In the current application, we will evaluate the peanut allergy inhibition in vivo using a humanized mice model. Simultaneously, we will exploit the potential of the cHBI as a platform to block other allergens including shellfish, dust mite and hazelnut. The proposed work is innovative and significant because it (i) offers a novel molecular design approach to inhibit “the key event” triggering an allergic response with potential long-term clinical applications in food, environmental and drug allergies, (ii) it does so without any non-specific suppression of immune system components, and (iii) develops a much needed, physiologically relevant, easily adjustable and reproducible platform (nanoallergens), which will be used in identifying immunodominant public allergen epitopes and their relative significance during degranulation responses. Lastly, this application has high potential impact because cHBI has the potential for translation to the clinic.