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

使用关联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：
9530416
负责人：
John Michael Streicher
金额：
$ 23.03万
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/9530416
关键词：
Absence of pain sensation Active Sites Acute Address Adverse effects 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 Mono-S Morphine Mus Naloxone Nociception Nylons Opioid Opioid Analgesics Opioid Receptor Pain Peptides Pharmaceutical Preparations Pharmacology Preparation Process Rewards Role Series Signal Transduction Site 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 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，包括对阿片类药物的耐受性和抗伤害感受。许多小组发现抑制通过各种手段的DOR降低了吗啡等MOR激动剂的副作用。而这种相互作用尚不清楚，一种很大的可能性是形成MOR-DOR异二聚体（MDOR）。许多组表明，MDOR可以在体外形成异源表达系统，具有独特的与单体形式相比，药理学和信号转导轮廓。 Devi及其同事最近开发了一种MDOR选择性抗体，并使用该抗体证明了MDOR上调长期用吗啡治疗的小鼠的大脑。其他实验表明MDOR促进耐受性，依赖性和在体内寻求药物。虽然已经开发了Mdor选择性激动剂，但没有曾经创建了已知的类似药物的拮抗剂，限制了我们确定的作用的能力 Mdor在体内。为了解决这一缺乏，我们创建了一系列新型潜在的选择性肽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）长度（15-42个原子）柔性聚酰胺间隔物的药效团。我们测试了这个初步使用放射性配体结合和35S-GTPγS在拮抗剂模式下使用MOR，DOR和 MDOR表达细胞系。我们发现了令人信服的证据，表明我们的初步系列有选择地针对 MDOR，最佳化合物24原子垫片长度的选择性比为〜91倍。由此建造最初的成功，我们旨在当前的提议探索我们的MDOR结构活性关系（SAR）复合系列，并通过调节药效团来提高MDOR的复合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），适度刚性[Gly-Pro]）。这些研究将在迭代开发过程中使用最佳化合物进行从每个系列中通知下一个系列，最小化复合号。在此SAR开发之后，我们将使用我们最潜在和选择性化合物开始在体内测试MDOR活动。这将完成在脑室内（ICV）和鞘内（IT）将MDOR拮抗剂注射到小鼠之前 MOR（DAMGO），DOR（DSLET）和MDOR（CYM51010）选择性激动剂引起的伤害感受。这些研究将证明我们体内化合物的选择性。我们还将用MDOR预处理小鼠在诱导耐受性和吗啡依赖性之前，拮抗剂开始探索体内作用这些阿片类药物副作用中的MDOR，文献提出了。在短期内，这个最初的优化系列将提供有用的工具来询问MDOR在体内的作用。长期修改改善可药用性（糖基化等），这些化合物可能为选择性提供基础 MDOR的靶向治疗以改善阿片类药物治疗的副作用特征。