Ghrelin Deacylase as a Treatment for Opioid Polysubstance Abuse

生长素释放肽脱酰酶治疗阿片类多物质滥用

• 批准号: 10510245
• 负责人: CHANG-GUO ZHAN
• 金额: $ 170.09万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10510245
• 关键词: Acute; Adverse effects; Affect; Aftercare; Alcohols; Amino Acids; Amphetamines; Animals; Attenuated; Behavior; Binding; Biological Assay; Blood Circulation; Brain; Butyrylcholinesterase; Chimeric Proteins; Chronic; Cocaine; DR1 gene; Data; Dependence; Development; Disease; Dopamine; Dopamine Receptor; Down-Regulation; Drug Exposure; Drug abuse; Enzymes; Esthesia; Euphoria; Feeling; Fentanyl; G-Protein-Coupled Receptors; Gastric Acid; Heroin; High Prevalence; Human; Hydrolase; Hyperactivity; Hypesthesia; In Vitro; Ingestion; Investigation; Lead; Ligands; Literature; Methamphetamine; Morphine; National Institute of Drug Abuse; Neurons; Nicotine; Octanoic Acids; Opioid; Pathway interactions; Peptides; Perception; Pharmaceutical Preparations; Play; Presynaptic Terminals; Process; Production; Property; Proteins; Rattus; Receptor Signaling; Recombinant Proteins; Relapse; Reporting; Research; Rewards; Rodent; Rodent Model; Role; Self Administration; Side; Signal Transduction; Site-Directed Mutagenesis; Somatotropin; Stimulant; Stimulus; Stomach; Structure; Substance Use Disorder; Substance of Abuse; Synapses; System; Testing; Therapeutic; Travel; Withdrawal; Work; animal data; antagonist; base; behavioral study; conditioned place preference; deacylation; design; dopamine system; dopamine transporter; dopaminergic neuron; drug reward; effective therapy; enzyme activity; ghrelin; growth hormone secretagogue receptor; improved; in vivo; in vivo evaluation; methamphetamine effect; methamphetamine use; mutant; novel; novel therapeutic intervention; novel therapeutics; opioid use disorder; opioid user; pleasure; polysubstance abuse; polysubstance use; protein expression; protein purification; rational design; receptor; receptor expression; response; substance use; substance use treatment; success; therapeutic candidate; treatment strategy

With currently available medications for opioid use disorder (OUD), the relapse rate is very high. Further, co- use of an opioid and a stimulant, such as methamphetamine (METH), undermines success in treatment for OUD. It is highly desired to develop novel therapeutic options for treatment of OUDs including polysubstance use disorders (PSUDs). Recently reported studies have revealed an interesting role of ghrelin in drug abuse and reward-relevant behaviors. Ghrelin is known as a “growth-hormone-releasing acylated peptide from stomach”, a 28 amino-acid peptide with the third residue (Ser3) acylated by n-octanoic acid. Ghrelin is produced in the stomach, travels to the brain through blood circulation, and acts on ghrelin (or growth hormone secretagogue) receptor (GHSR) to stimulate the mesolimbic dopamine reward pathway and increase rewarding behaviors in rodents. GHSR antagonism has been shown to attenuate rewarding effects induced by various substances including METH, amphetamine, fentanyl, morphine, heroin, cocaine, nicotine, and alcohol. For example, administration of a selective GHSR antagonist significantly reduced the fentanyl‐seeking/relapse‐like behavior in rats. On the other hand, GHSR has diverse regulatory roles associated with its constitutive activity (the activity in the absence of ghrelin ligand) and, hence, GHSR antagonism could also result in unwanted adverse effects. Thus, alternative strategies targeting ghrelin itself could be more interesting. However, whether targeting ghrelin itself would be effective to attenuate the drug rewarding effects remains controversial in literature as ghrelin levels before and after treatment were not measured in previous animal studies targeting ghrelin itself. It is unclear whether any of the previously used approaches was able to decrease the ghrelin level significantly enough to attenuate the substance reward. We propose to use our recently identified ghrelin deacylase (a mutant of human butyrylcholinesterase or BChE) as a safe and effective ghrelin modulator to attenuate substance rewarding effects. Such a ghrelin modulator is expected to be as effective as a GHSR antagonist in treatment of substance use disorders (SUDs) including PSUDs, but without interacting with any brain receptors/transporters. Specifically, in this investigation, we will first evaluate the BChE mutant for its in vivo potency in modulating ghrelin and attenuating the rewarding and reinforcing effects of representative opioids (fentanyl and heroin) and their combinations with METH in various rodent models. Then, we will design and discover a new BChE mutant as a more potent ghrelin deacylase with further improved catalytic activity for ghrelin diacylation, develop its long-acting fusion protein form, and examine the long-term in vivo effects of the long-acting ghrelin deacylase. Accomplishment of this investigation will determine whether ghrelin itself is a truly effective target and whether a potent ghrelin deacylase as a ghrelin modulator is truly effective for treatment of PSUDs. If the answers are all positive, the most effective long-acting ghrelin deacylase (recombinant protein) to be tested and developed may also serve as a promising therapeutic candidate for treatment of PSUDs.

借助目前可用于用于绿化疾病的药物（OUD），继电器速率非常高。此外，共同 使用阿片类药物和刺激剂，例如甲基苯丙胺（METH），破坏了OUD治疗的成功。 非常希望开发新颖的治疗选择来治疗Oud，包括使用PolySubstance 疾病（psuds）。最近报道的研究表明，生长素释放肽在药物滥用和 与奖励相关的行为。生长素素被称为“从Stalloch产生的生长激素释放酰化肽”，A 28氨基酸胡椒粉，其第三居住地（Ser3）被N-辛酸酰化。生长素是在 胃，通过血液循环向大脑传播，并作用于生长素（或成长的霍斯纳分泌物） 受体（GHSR）刺激中比明胺奖励途径并增加奖励行为 啮齿动物。 GHSR拮抗作用已被证明可以减弱各种物质引起的奖励效应 甲基苯丙胺，苯丙胺，芬太尼，吗啡，海洛因，可卡因，尼古丁和酒精。例如， 选择性GHSR拮抗剂的给药可显着降低寻求芬太尼/复发的行为 在老鼠中。另一方面，GHSR具有与其本构活性相关的潜水调节作用（该活动 在没有生长素蛋白配体的情况下），因此，GHSR拮抗作用也可能导致不良反应。 这是针对生长素本身的替代策略可能会更有趣。但是，是否针对生长素 在ghrelin中，衰减药物奖励效果的衰减本身将有争议 在以前针对生长素蛋白本身的动物研究中，未测量治疗前后的水平。这是 尚不清楚任何先前使用的方法是否能够显着降低生长素素水平 足以减轻物质奖励。我们建议使用我们最近鉴定出的生长素蛋白脱酰基酶（突变体 人丁酰胆碱酯酶或BCHE）作为安全有效的生长素调节剂，可衰减底物 奖励效果。预计这种生长素蛋白调节剂在治疗中与GHSR拮抗剂一样有效 物质使用障碍（SUD），包括psuds，但没有与任何大脑受体/转运蛋白相互作用。 具体而言，在这项调查中，我们将首先评估BCHE突变体的体内效力 生长素并衰减代表性阿片类药物（芬太尼和海洛因）的奖励和增强作用 它们与各种啮齿动物模型中的甲基苯丙胺组合。然后，我们将设计并发现一个新的BCHE突变体 作为具有进一步提高催化活性的尿素二酰基化催化活性的较高潜在的生长素蛋白脱酰基酶，发展其发展 长效融合蛋白形式，并检查长效生长素蛋白脱酰基酶的长期体内效应。 完成这项调查将确定Ghrelin本身是否是真正有效的目标，以及是否是否 潜在的生长素蛋白脱酰基酶作为生长素蛋白调节剂对PSUD的治疗确实有效。如果答案都是 阳性，最有效的长作用生长素蛋白脱酰基酶（重组蛋白）要测试和开发 还可以作为PSUD治疗的有前途的治疗候选者。