Development of a Selective Mu-Delta Opioid Receptor Heterodimer Antagonist Using a Linked Bivalent Pharmacophore Approach

使用链接二价药效团方法开发选择性 Mu-Delta 阿片受体异二聚体拮抗剂

基本信息

批准号：
9926491
负责人：
John Michael Streicher
金额：
$ 0.79万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9926491
关键词：
Absence of pain sensation Active Sites Acute Address Affect Affinity Agonist Antibodies Avidity Behavioral Binding Binding Sites Biological Assay Brain Cell Line Chinese Hamster Ovary Cell Chronic Complex Coupling DSLET Data Dependence Development Dose Entropy Female G-Protein-Coupled Receptors Heterodimerization Immune response Implant In Vitro Individual Intrathecal Injections Lead Length Ligand Binding Ligands Link Literature Measures Mediating Membrane Methodology Modification Morphine Mus Naloxone Nociception Nylons Opioid Opioid Analgesics Opioid Receptor Pain Peptides Pharmaceutical Preparations Pharmacology Preparation Process Rewards Role Series Signal Transduction Site Structure Structure-Activity Relationship System Tail Testing Therapeutic Up-Regulation Water Withdrawal base delta opioid receptor design drug seeking behavior experimental study flexibility glycosylation glycylproline improved in vitro Model in vivo in vivo evaluation male member novel opiate tolerance opioid therapy pharmacophore prolyl-glycyl-glycine prolyl-tyrosine radioligand receptor response reverse tolerance side effect success targeted treatment therapeutic evaluation therapeutic target tool treatment duration tyrosyl-proline

项目摘要

The mu and delta opioid receptors (MOR, DOR) modulate many of the same brain processes in vivo, including tolerance and anti-nociception in response to opioid drugs. Many groups have found that inhibiting the DOR through various means decreases the side effects of MOR agonists like morphine. While the basis for this interaction is unknown, one strong possibility is the formation of a MOR-DOR heterodimer (MDOR). Many groups have shown that the MDOR can form in heterologous expression systems in vitro, with a unique pharmacology and signal transduction profile when compared to the monomeric forms. Devi and colleagues recently developed an MDOR selective antibody, and used this antibody to demonstrate MDOR upregulation in the brains of mice chronically treated with morphine. Other experiments suggested that the MDOR promotes tolerance, dependence, and drug seeking in vivo. While MDOR selective agonists have been developed, no known drug-like antagonist has ever been created to our knowledge, limiting our ability to determine the role of MDOR in vivo. To address this lack, we created a novel series of potential selective peptide MDOR antagonists by connecting low affinity MOR (H-Tyr-Pro-Phe-D1Nal-NH2) and moderate affinity DOR (Tyr-Tic- OH) pharmacophores with a variable length (15-42 atom) flexible polyamide spacer. We tested this preliminary series in vitro using radioligand binding and 35S-GTPγS coupling in antagonist mode using MOR, DOR, and MDOR expressing cell lines. We found compelling evidence that our preliminary series selectively targets the MDOR, with a selectivity ratio of ~91 fold for our best compound, the 24 atom spacer length. Building from this initial success, we aim in the current proposal to explore the MDOR structure-activity relationship (SAR) of our compound series, and improve compound potency and selectivity at the MDOR by modulating pharmacophore affinity (increased MOR, decreased DOR) as well as linker rigidity (minimally rigid [gly-gly-pro], moderately rigid [gly-pro]). These studies will be performed in an iterative development process, using the best compound from each series to inform the next series, minimizing compound number. After this SAR development, we will use our most potent and selective compound to begin to test MDOR activity in vivo. This will be accomplished by intracerebroventricular (icv) and intrathecal (it) injection of MDOR antagonist into mice prior to tail-flick anti- nociception evoked by MOR (DAMGO), DOR (DSLET), and MDOR (CYM51010) selective agonists. These studies will demonstrate the selectivity of our compound in vivo. We will also pre-treat mice with MDOR antagonist prior to the induction of tolerance and dependence with morphine to begin to explore the in vivo role of the MDOR in these opioid side effects, which has been suggested by the literature. In the short term, this initial optimized series will provide a useful tool to interrogate the role of the MDOR in vivo. Long term, through modifications to improve drugability (glycosylation, etc.), these compounds may provide the basis for selective MDOR targeted therapeutics to improve the side effect profile of opioid therapy.

mu 和 delta 阿片受体（MOR、DOR）在体内调节许多相同的大脑过程，许多研究小组发现，阿片类药物具有抑制作用，包括耐受性和抗伤害性。 DOR 通过多种方式降低 MOR 激动剂（如吗啡）的副作用。这种相互作用是未知的，一种强烈的可能性是形成 MOR-DOR 异二聚体 (MDOR)。研究小组表明，MDOR 可以在体外异源表达系统中形成，具有独特的与单体形式相比，药理学和信号转导特性。最近开发了一种 MDOR 选择性抗体，并使用该抗体证明了 MDOR 上调其他实验表明，MDOR 可以促进长期接受吗啡治疗的小鼠的大脑。耐受性、依赖性和体内药物寻求虽然已经开发出 MDOR 选择性激动剂，但还没有。据我们所知，已知的药物样拮抗剂已经被创造出来，限制了我们确定其作用的能力为了解决这一缺陷，我们创建了一系列新型潜在选择性肽 MDOR。通过连接低亲和力 MOR (H-Tyr-Pro-Phe-D1Nal-NH2) 和中等亲和力 DOR (Tyr-Tic- OH）具有可变长度（15-42 个原子）柔性聚酰胺间隔基的药效团我们对此进行了初步测试。使用 MOR、DOR 和拮抗剂模式使用放射性配体结合和 35S-GTPγS 偶联进行体外系列研究我们发现了令人信服的证据，表明我们的初步系列选择性地针对 MDOR 表达细胞系。 MDOR，我们最好的化合物的选择性比约为 91 倍，即由此构建的 24 个原子间隔基长度。初步成功后，我们的目标是在当前的提案中探索我们的 MDOR 结构-活性关系（SAR）化合物系列，并通过调节药效团提高化合物在 MDOR 的效力和选择性亲和力（MOR 增加，DOR 减少）以及接头刚性（最小刚性 [gly-gly-pro]，中等刚性）这些研究将使用最好的化合物在迭代开发过程中进行。在此 SAR 开发之后，我们将从每个系列中得知下一个系列，从而最大限度地减少化合物数量。使用我们最有效和最具选择性的化合物开始测试体内 MDOR 活性。通过在甩尾之前向小鼠脑室内（icv）和鞘内（it）注射MDOR拮抗剂 MOR (DAMGO)、DOR (DSLET) 和 MDOR (CYM51010) 选择性激动剂引起的伤害。研究将证明我们的化合物在体内的选择性。我们还将用 MDOR 预处理小鼠。在诱导吗啡耐受和依赖性之前使用拮抗剂，开始探索其体内作用这些阿片类药物副作用中的 MDOR 的影响，这在短期内是由文献提出的。初始优化系列将提供一个有用的工具来探究 MDOR 在体内的长期作用。修饰以提高成药性（糖基化等），这些化合物可能为选择性 MDOR 靶向治疗可改善阿片类药物治疗的副作用。