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: 8092919
• 负责人: PAUL WENTWORTH
• 金额: $ 23.69万
• 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 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8092919
• 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; 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）：PA-10-069 下的 R21 提案是免疫药物疗法和 NIDA 滥用药物疫苗计划的一次飞跃，并“测试了一个新颖且重要的假设，如果通过实验证实，该假设将具有重大意义”对小分子疫苗思维的影响”。该核心平台利用了新发现的能力，可以用 PI 研究中的功能性酰肼分子对天然状态的补体蛋白 C3 进行化学修饰。该小组以小分子滥用药物分子作为与 C3 复合的免疫原，生产革命性疫苗。疫苗开发的四步理论方法如图 1 所示。这个概念非常简单，在全球范围内适用，并且具有潜在的突破性。免疫原是宿主蛋白（非外来蛋白），不需要外部佐剂。这种新的免疫原平台的开发和应用将在以下两个目标的范围内进行测试： 目标 1 线性 B 细胞表位-小分子酰肼的设计、合成和连接至鼠 C3（图 1 中的步骤 1-3） 。将研究这种新的免疫药物治疗平台主要应用于可卡因（COC），然后应用于小鼠中的苯环己哌啶（PCP）和甲基苯丙胺（MET）。因此，将使用 PI 实验室的技术例程分离和纯化小鼠 C3（非人类），用于该过程的第 1 步。对于第 2 步，将在 PI 实验室中合成包含 Mengo 病毒 VP1259-277 线性 B 细胞表位的两个酰肼炔，并且这些 B 细胞表位酰肼炔将使用 PI 中已开发的条件与天然鼠 C3 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 实验室中合成，并将使用标准“点击”化学条件与第 2 步中的 C3-炔烃缀合物发生反应。目标 2. 使用 C3 药物生物共轭物对小鼠进行主动免疫并对免疫反应进行定量（图 1 中的步骤 4）。来自 Aim 1 的 C3-生物缀合物将用于在存在和不存在外部佐剂 (RIBI) 的情况下对雌性 Balb/c 小鼠（每组 3-5 只）进行免疫。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 持久性均被测量。将使用药物的 KLH 缀合物进行比较免疫接种作为对照。 公共卫生相关性：生产允许免疫系统识别小分子的有效疫苗的能力是目前挖掘滥用科学中尚未满足的需求。该提案利用先天免疫系统的力量，通过化学合成与补体蛋白 C3 偶联的小分子疫苗来产生新型小分子疫苗。