C3 chemical conjugation to improve the anti-cocaine immune response

C3化学缀合改善抗可卡因免疫反应

• 批准号: 8233313
• 负责人: PAUL WENTWORTH
• 金额: $ 28.43万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8233313
• 关键词: Active Immunization; Adjuvant; Affinity; Alkynes; Antigens; Azides; B-Lymphocyte Epitopes; Binding; Chemicals; Chemistry; Cocaine; Complement; Complex; Coupled; Development; Female; Human; Immune response; Immune system; Immunization; Immunoglobulin G; Keyhole Limpet Hemocyanin; Laboratories; Measures; Mengovirus; Methamphetamine; Mus; Pharmaceutical Preparations; Phencyclidine; Plasma; Process; Proteins; Research; Science; Serum; Specificity; Techniques; Testing; Thinking; Vaccines; analog; base; chemical synthesis; comparative; design; drug abuser; drug of abuse; improved; novel; programs; public health relevance; research study; small molecule; vaccine development

DESCRIPTION (provided by applicant): This R21 proposal under PA-10-069 offers to be a leap-forward in immunopharmacotherapy and NIDA's vaccine program for drugs of abuse and 'tests a novel and significant hypothesis which if confirmed by experiment would have a substantial impact on thinking regarding vaccines against small molecules'. The core platform exploits the newly discovered ability to chemically modify complement protein C3 in its native state with functional hydrazide molecules that was made in the PI's research. group to generate revolutionary vaccines based on small molecule drug of abuse molecules as immunogens complexed to C3. The four step theoretical approach to vaccine development is shown in Figure 1. The concept is at once simple, globally applicable and potentially ground breaking. The immunogen is a host protein (not foreign) and should not require an external adjuvant. The development and application of this new immunogen platform will be tested under the remit of the following two aims: Aim 1 Design, synthesis and attachment of linear B-cell epitope-small molecules hydrazides to murine C3 (STEPS 1-3 of Figure 1). The aplication of this new immunopharmacotherapy platform primarily to cocaine (COC), and then to phencyclidine (PCP) and methamphetamine (MET) in mice will be investigated. Thus, mouse C3 (not human) will be isolated and purified for Step 1 of the process using a technique routine in the PI's lab. For Step 2, two hydrazide alkynes, incorporating the Mengo virus VP1259-277 linear B-cell epitope, will be synthesized in the PI's laboratory and these B-cell epitope hydrazide alkynes will be reacted with native murine C3 using conditions already developed in the PI's laboratory. For Step 3, azide analogs of cocaine, PCP and methamphetamine will be synthesized in the PI's laboratory and will be reacted with the C3-alkyne conjugates from Step 2 using standard 'click' chemistry conditions. Aim 2. Active immunization of mice with C3-drug bioconjugates and quantification of the immune response (STEP 4 of Figure 1). The C3-bioconjugates from Aim 1 will be used to immunize female Balb/c mice (3-5 per group), in the presence and absence of external adjuvant (RIBIs). Anti-drug serum titers, polyclonal IgG binding affinity, binding specificity (drug versus B-cell epitope), and IgG persistence all being measured. Comparative immunizations with KLH-conjugates of the drugs will be performed as a control. PUBLIC HEALTH RELEVANCE: The ability to produce effective vaccines that allow immune system recognition of small molecules is a current unmet need in dug abuse sciences. This proposal harnesses the power of the innate immune system by chemical synthesis of small molecule vaccines coupled to complement protein C3 to generate novel small molecule vaccines.

描述（由申请人提供）：根据PA-10-069下的R21提案提议是免疫药物治疗和NIDA的滥用药物疫苗计划的跃迁，并检验了一种新颖而重要的假设，如果通过实验证实，这将具有实验性的实质性。对针对小分子的疫苗思考的影响。核心平台利用了新发现的能力，可以通过PI研究中制作的功能性夹层分子化学修改其本地蛋白质C3的补体C3。基于滥用分子的小分子药物，将革命性疫苗产生，因为免疫原子与C3复合。疫苗开发的四个步骤理论方法如图1所示。该概念既简单，全球适用且有潜在的突破。免疫原是一种宿主蛋白​​（不是异物），不需要外部佐剂。该新的免疫原平台的开发和应用将在以下两个目的的位置下进行测试：AIM 1设计，线性B-cell表位表位分子hidyazides hildazides hildrazides to Murine C3（图1的步骤1-3） 。将研究这种新的免疫药物治疗平台，主要是可卡因（COC），然后研究小鼠中苯基二肽（PCP）和甲基苯丙胺（MET）。因此，将使用PI实验室中的技术例程将小鼠C3（不是人）分离并纯化为该过程的步骤1。对于步骤2，将在PI的实验室中合成两个夹层藻类，其中包含Mengo病毒VP1259-277线性B细胞表位，这些B细胞表位hirdrazide Alkynes将使用PI在PI的条件中与天然鼠类C3进行反应实验室。对于第3步，可卡因，PCP和甲基苯丙胺的叠氮化物类似物将在PI的实验室中合成，并将使用标准“点击”化学条件与C3-Alkyne结合物反应。 AIM 2。用C3-毒物生物缀合物对小鼠进行主动免疫和免疫反应的定量（图1的步骤4）。在存在和不存在外部佐剂（ribis）的情况下，来自AIM 1的C3-Bioconjugates将用于免疫雌性BALB/C小鼠（每组3-5）。抗药物血清滴度，多克隆IgG结合亲和力，结合特异性（药物与B细胞表位）和IgG持久性均可测量。用药物的KLH偶联物进行比较免疫将作为对照进行。 公共卫生相关性：生产有效疫苗以允许免疫系统识别小分子的能力是当前未满足的滥用科学的需求。该提议通过化学合成的小分子疫苗结合以补充蛋白C3来生成新型的小分子疫苗，从而利用了先天免疫系统的能力。