Synthesis and in vitro and in vivo screening of fused and tethered heterocyclic peptidomimetics for the discovery of new analgesics with decreased side effects

融合和束缚杂环肽模拟物的合成以及体外和体内筛选，以发现副作用减少的新型镇痛药

• 批准号: 10297832
• 负责人: Jay P. McLaughlin
• 金额: $ 29.71万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-11 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10297832
• 关键词: Absence of pain sensation; Acetic Acids; Address; Affinity; Agonist; Amino Acids; Analgesics; Area; BCL2L11 gene; Behavior assessment; Binding; Binding Proteins; Biological; Biological Assay; Biological Availability; Blood; Blood - brain barrier anatomy; Brain; Clinical; Constipation; Cyclic AMP; Data; Development; Dose; Drug Kinetics; Evaluation; Exhibits; Failure; Generations; Goals; Harvest; Heterocyclic Compounds; Imidazolidines; Imines; In Vitro; Lead; Lead levels; Libraries; Liver; Mass Spectrum Analysis; Measurement; Mediating; Metabolic; Molecular Probes; Morphine; Motor Activity; Mus; Naloxone; Opiate Addiction; Opioid; Opioid Analgesics; Opioid Antagonist; Opioid Receptor; Opioid agonist; Oral; Oral Administration; Outcome; Pain; Parents; Peptides; Peripheral; Permeability; Physiological; Piperazines; Plasma Proteins; Property; Research; Rewards; Sedation procedure; Series; Side; Signal Transduction; Suggestion; System; Tail; Testing; Time; Ventilatory Depression; Vertebral column; Water; Withdrawal; abuse liability; antagonist; aqueous; base; beta-arrestin; computational chemistry; conditioned place preference; design; gastrointestinal epithelium; imidazolone; improved; in vivo; innovation; interest; intravenous administration; kappa opioid receptors; liquid chromatography mass spectrometry; novel; peptidomimetics; radioligand; receptor; recruit; respiratory; scaffold; screening; side effect; small molecule; therapeutic opioid

We hypothesize that the development of peripherally-restricted, mixed-activity opioid receptor agonists will produce robust analgesia while exhibiting reduced side effects, including a lack of respiratory depression or reinforcing properties. In preliminary studies, the in vitro screening of novel bis-imidazolidines and fused heterocyclic lead compounds BIM-22 and FDC-14 identified mixed-opioid receptor agonist activity. After peripheral (i.p.) administration to mice, both compounds demonstrated antinociception equivalent to morphine in the 55oC warm-water tail withdrawal (tail-flick) assay, but FDC-14 was 25 times more potent than morphine in the acetic acid writhing test, with an antinociceptive potency ratio of 72.7, suggestive of peripherally- restricted activity. Confirming this, LC-MS/MS studies detected BIM-22 and FDC-14 in harvested mouse blood, but not brain, after i.v. administration. This proposal addresses two research areas of particular interest for this FOA: i) The development of new and innovative molecular probes for receptors and new strategies for innovative peptidomimetic design, and ii) The identification of structurally diverse, orally active, metabolically stable peripherally-restricted opioid agonists. We propose six interacting aims: In Aim 1, we propose the computationally guided synthesis of heterocyclic peptidomimetics: bis-imidazolidin-2-imines, piperazines, bis- piperazine, bis-imidazolone and fused heterocyclic libraries. In Aim 2, we will screen all compounds in competition radioligand binding assays to determine affinity for μ- (MOR), δ- (DOR), and κ- (KOR) opioid receptors, and in functional assays to identify dual δ/κ or dual δ/µ agonists. In Aim 3, ten selected compounds will be evaluated in vitro for pharmacokinetic properties and screened in vivo for antinociception using the mouse tail-flick assay. Lead compounds identified from this aim will guide 2nd generation SAR studies to enhance peripherally-selective activity. In Aim 4: we will perform full in vivo antinociceptive characterization of lead agonist compounds. The most stable, active 4 agonists not crossing the BBB in Aims 2+3 will be evaluated after i.p. administration with mouse tail-flick and acetic-acid writhing assays for efficacy, duration of action, and opioid receptor selectivity. In Aim 5: we will characterize the bioavailability in mice of the selected 4 agonists following oral administration and confirm their inability to penetrate the blood brain barrier. In Aim 6: The two most potent bioavailable novel agonists identified in Aims 1-5 will be examined for liabilities, specifically antinociceptive tolerance, respiratory and hyperlocomotor effects in the CLAMS physiological and behavioral assessment system, sedation and disruption of coordinated locomotor activity in the rotorod assay, and assessment for rewarding or aversive effects in the conditioned place preference (CPP) assay. Effects on GI transit will also be evaluated to assess effects on constipation. In summary, we expect to generate novel peripherally-restricted, mixed-activity peptidomimetic opioid receptor agonists as both probes and analgesics with reduced side effects, thereby significantly impacting analgesic development.

我们假设外周限制的混合活性阿片受体激动剂的发展将 在副作用降低时产生健壮的镇痛作用，包括缺乏呼吸道抑郁或 加强特性。在初步研究中，新型双咪唑啉和融合的体外筛查 杂环铅化合物BIM-22和FDC-14鉴定出混合阿片受体激动剂活性。后 外围（i.p.）给小鼠给药，两种化合物均表现出与吗啡相等的抗伤害感受 在55oC温暖的水尾撤离（尾式）测定中，FDC-14的潜力是吗啡的25倍 在乙酸写作测试中，抗感染性效力比为72.7 受限活动。确认这一点，LC-MS/MS研究在收获的小鼠血液中检测到BIM-22和FDC-14， 但不是大脑，静脉注册之后行政。该提案针对这两个研究领域特别感兴趣 FOA：i）开发新的和创新的分子问题，用于受体和新策略的新策略 创新的肽概性设计，ii）结构上的潜水员，口服活性，代谢 稳定的外围限制性阿片类药物激动剂。我们提出了六个互动目的：在AIM 1中，我们提出了 杂环辣椒材料的计算指导性合成：bis-imidazolidin-2- imines，哌嗪，bis- 哌嗪，二咪唑酮和融合的杂环库。在AIM 2中，我们将筛选所有化合物 竞争放射线结合的主张，以确定对μ-（MOR），δ-（DOR）和κ-（kor）阿片类药物的亲和力 受体，以及在功能测定中识别双δ/κ或双δ/μ激动剂。在AIM 3中，十种选择的化合物 将在体外评估药代动力学特性，并在体内筛选使用抗伤害感受 鼠标尾灯测定法。从此目标中确定的铅化合物将指导第二代SAR研究 增强外围选择性活性。在AIM 4中：我们将执行完整的体内抗伤害感受性表征 铅激动剂化合物。最稳定的，最稳定的4个激动剂未在目标2+3中越过BBB 在腹腔内评估用小鼠尾灯和乙酸酸的效率进行管理，持续时间 动作和阿片类药物的选择性。在AIM 5中：我们将表征所选4的小鼠的生物利用度 口服给药后的激动剂并确认其无法穿透血脑屏障。在AIM 6中： 在目标1-5中确定的两个最潜在的生物利用新颖的激动剂将用于负债， 特定于蛤lam生理和 行为评估系统，镇静和在Rotorod测定中协调的运动活性的破坏， 以及评估条件地点偏好（CPP）测定中的奖励或厌恶作用。影响 GI Transit还将评估以评估对便秘的影响。总而言之，我们希望产生小说 外围限制性的，混合活性辣椒imimetic ipio-topior受体激动剂是问题和镇痛药 副作用降低，从而显着影响镇痛的发育。

项目成果

Bis-Cyclic Guanidine Heterocyclic Peptidomimetics as Opioid Ligands with Mixed μ-, κ- and δ-Opioid Receptor Interactions: A Potential Approach to Novel Analgesics.

• DOI: 10.3390/ijms23179623
• 发表时间: 2022-08-25
• 期刊: International journal of molecular sciences
• 影响因子: 5.6