Development of opioid and ketamine probes for in vivo photopharmacology

用于体内光药理学的阿片类药物和氯胺酮探针的开发

• 批准号: 10401573
• 负责人: Matthew R. Banghart
• 金额: $ 190.58万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10401573
• 关键词: Absence of pain sensation; Animals; Antidepressive Agents; Basic Science; Behavioral; Behavioral Assay; Biological Assay; Brain; Chemistry; Clinical; Communities; Consultations; Cultured Cells; Dendritic Spines; Development; Diffusion; Dissociative Anesthetics; Dopamine; Drug Kinetics; Drug usage; Electrophysiology (science); Exhibits; Family; Fiber; Fiber Optics; Fluorescent Probes; Generations; Goals; Image; Implant; In Vitro; Ketamine; Lead; Light; Link; Location; Measurement; Methods; Mus; N-Methyl-D-Aspartate Receptors; Naloxone; Nervous system structure; Neurophysiology - biologic function; Opioid; Opioid Analgesics; Opioid Peptide; Opioid Receptor; Optics; Outcome; Oxymorphone; Pain; Pain Disorder; Penetrance; Penetration; Performance; Pharmaceutical Preparations; Pharmacology; Photometry; Photons; Photophobia; Phototherapy; Prefrontal Cortex; Prodrugs; Property; Reagent; Research; Resolution; Rewards; Risk; Route; Signal Transduction; Site; Slice; Specificity; Spinal Cord; Stimulus; Structure; Therapeutic; Time; Toxic effect; Ultraviolet Rays; Variant; Work; analog; antagonist; awake; base; behavioral response; blood-brain barrier penetration; brain tissue; design; drug action; experience; fluorophore; improved; in vivo; insight; minimally invasive; neurobiological mechanism; neurochemistry; neuromechanism; novel; opioid use; optical fiber; photoactivation; photolysis; receptor; relating to nervous system; side effect; small molecule; spatiotemporal; subcutaneous; tool; two-photon; uptake; water solubility

Project Summary Pharmacological probes are widely used to study the nervous system. Despite often exhibiting exquisite specificity for target receptors, due to diffusion, traditional small molecule drugs act slowly and with spatial imprecision. This impedes neuropharmacological studies in vivo, particularly those involving high-resolution electrophysiological, imaging, and behavioral tracking methods. Such studies greatly benefit from the ability to correlate measurements with well-defined, time-locked stimuli that can be readily varied in intensity and duration. To meet this need, we are developing “caged” drugs that can be applied systemically in an inactive form and subsequently released in the brain with high spatial and temporal precision using short light flashes. To enable compatibility with optical measurements of neural function involving fluorescent probes, we will develop new photochemical protecting groups that are wavelength tuned to be spectrally orthogonal to common green and red fluorophores, in terms of both one-photon and two-photon excitation. We will further optimize these “caging” groups to facilitate the brain penetrance of the resulting caged drugs. We will evaluate the utility of new caging groups by incorporating them into 2nd generation caged opioid drugs that should provide significant experimental advantages over our 1st generation variants, which could only be photoactivated with ultraviolet light. We will rigorously validate new caged opioid drugs using in vitro, ex vivo, and in vivo experimental paradigms, culminating in behavioral assays and fiber photometry recordings of opioid-evoked neurochemical signaling. In addition, we will develop caged ketamine derivatives that can be used to study the neural mechanisms underlying dissociative states, as well as ketamine’s rapid antidepressant actions. Caged ketamine variants will also be evaluated using in vitro, ex vivo, and in vivo experimental paradigms, including measurements of spinogenesis and neural activity in the prefrontal cortex. These efforts will involve the development of an optical configuration for simultaneous two-photon imaging and one photon photolysis through implanted prisms. To maximize end-user uptake, performance criteria for both caged drug families are determined through extensive consultation with the scientific community.

项目摘要 药理学探针被广泛用于研究神经系统。尽管经常表现出对靶受体的精确特异性，但由于扩散，传统的小分子药物作用缓慢且具有空间不精确性。这阻碍了体内神经药理学研究，特别是那些涉及高分辨率电生理学，成像和行为跟踪方法的研究。这些研究极大地受益于将测量与定义明确的、时间锁定的刺激相关联的能力，这些刺激可以在强度和持续时间上容易地变化。为了满足这一需求，我们正在开发“笼”药物，这些药物可以以非活性形式全身应用，随后使用短闪光以高空间和时间精度在大脑中释放。为了能够与涉及荧光探针的神经功能的光学测量兼容，我们将开发新的光化学保护基团，其波长调谐为在单光子和双光子激发方面与常见的绿色和红色荧光团在光谱上正交。我们将进一步优化这些“笼”组，以促进所得笼药物的脑外转运。我们将通过将其纳入第二代笼状阿片类药物中来评估新笼状基团的效用，这些药物应该比我们的第一代变体提供显着的实验优势，第一代变体只能用紫外线光活化。我们将严格验证新的笼阿片类药物在体外，离体和体内实验范例，最终在阿片类药物诱发的神经化学信号的行为测定和纤维光度记录。此外，我们还将开发笼状氯胺酮衍生物，可用于研究解离状态下的神经机制，以及氯胺酮的快速抗抑郁作用。还将使用体外、离体和体内实验范例评价笼式氯胺酮变体，包括测量前额叶皮层中的棘突发生和神经活动。这些努力将涉及开发一种光学配置，用于通过植入棱镜同时进行双光子成像和单光子光解。为了最大限度地提高最终用户的使用率，通过与科学界的广泛协商，确定了两种笼内药物系列的性能标准。