Development of a Novel Cell Based PDE10A Assay for Antipsychotic Drug Discovery

开发用于抗精神病药物发现的新型细胞 PDE10A 检测方法

基本信息

批准号：
8337753
负责人：
Wenshan Hao
金额：
$ 31.98万
依托单位：
CODEX BIOSOLUTIONS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-23 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8337753
关键词：
Adenylate Cyclase Adopted Adverse effects Affect Agonist Animal Model Antipsychotic Agents Area Behavioral Medicine Biochemical Biological Assay Biosensor Boxing Brain Cell Line Cells Chronic Clinical Clone Cells Complement Consult Coupled Cyclic AMP Cyclic GMP Detection Development Disease Dopamine D2 Receptor Drug Delivery Systems Drug Design Drug Industry Engineering Fluorescence Fluorescent Dyes Forskolin Funding G Protein-Coupled Receptor Genes General Population Genes Grant Individual Industry Inflammation Inhibitory Concentration 50 Knock-out Libraries Life Malignant Neoplasms Maryland Mental disorders Metabolism Neurobehavioral Manifestations New Jersey Nucleotides PDE4B Papaverine Performance Pharmaceutical Preparations Pharmacologic Substance Phase Process Property Protein Isoforms Protocols documentation Psychiatry Reader Receptor Cell Relapse Research Research Personnel Resources Schizophrenia Screening procedure Signal Transduction Small Business Innovation Research Grant Structure Symptoms Technology Therapeutic Thyrotropin Receptor Time Treatment Protocols United States Universities Variant West Virginia Zinc Fingers base cyclic-nucleotide gated ion channels drug candidate drug discovery high throughput screening improved inhibitor/antagonist innovation knockout gene novel nuclease phosphoric diester hydrolase practical application prevent small molecule success therapeutic development tool

项目摘要

DESCRIPTION (provided by applicant): Schizophrenia is a chronic and devastating mental disorder that affects 0.5-0.8 percent general population and over 2 million people in the United States. The current treatment regimen depends on D2 dopamine receptor-targeted drugs that are partially effective on symptoms and cause severe side effects, leading to high treatment discontinuation and relapse. Phosphodiesterase 10A (PDE10A) is emerging as a novel drug target for schizophrenia that promises the development of therapeutics with improved efficacy and less side effects. To develop a pipeline of PDE10A inhibitors, pharmaceutical and biotech companies are depending heavily on cell-free enzymatic assays and structure-based drug design. Although cell-based high-throughput screening (HTS) assays for PDE10A would greatly accelerate the drug discovery process and assist in discovering drug candidates with diverse mechanisms of actions and better pharmacological properties, there are no currently commercial products available to the pharmaceutical industry. Codex Biosolutions has a proprietary cAMP biosensor technology (ACTOne") that has been successfully developed into a cell-based assay for screening PDE4 inhibitors in a 1536-well HTS campaign. In this SBIR phase I funding, our objective is to extend the initial success to establish a PDE10A cell-based assay that will be used by the industry to develop innovative therapeutics for schizophrenia disease. We will conduct all the research in our facilities located in Maryland and New Jersey, which are equipped with all necessary resources to complete the project. We will also have Dr. Hanting Zhang from Departments of Behavioral Medicine & Psychiatry at West Virginia University, an expert on PDE research, to consult us on the project. We propose the following four specific aims for SBIR Phase I. 1. Generate stable HEK293-CNG-TSHR-PDE10A cell line. 2. Knock out PDE4D and PDE4B genes to generate HEK293-CNG-TSHR-PDE10A-PDE4D-/--PDE4B-/- cell line (PDE10A cell line). 3. Adapt the PDE10A cell line to the 384-well HTS format. 4. Perform high-throughput screening of the compound library to evaluate the assay performance in identifying known PDE10A inhibitors. The successful completion of Phase I will pave the way to extend the same technology to other PDE isoforms, leading to the establishment of a repertoire of selective cell-based assays for individual PDE variants that will serve drug discovery projects targeting broad therapeutic areas including CNS, inflammation, metabolism, and cancer.

描述（由申请人提供）：精神分裂症是一种慢性、毁灭性的精神障碍，影响美国 0.5-0.8% 的总人口，超过 200 万人。目前的治疗方案依赖于 D2 多巴胺受体靶向药物，这些药物对症状部分有效，但会引起严重的副作用，导致治疗中断和复发率很高。磷酸二酯酶 10A (PDE10A) 正在成为精神分裂症的新型药物靶点，有望开发出疗效更高、副作用更少的治疗方法。为了开发一系列 PDE10A 抑制剂，制药和生物技术公司在很大程度上依赖于无细胞酶测定和基于结构的药物设计。尽管基于细胞的 PDE10A 高通量筛选 (HTS) 检测将大大加速药物发现过程，并有助于发现具有不同作用机制和更好药理学特性的候选药物，但目前制药行业还没有可用的商业产品。 Codex Biosolutions 拥有专有的 cAMP 生物传感器技术 (ACTOne")，该技术已成功开发为基于细胞的检测方法，用于在 1536 孔 HTS 活动中筛选 PDE4 抑制剂。在本次 SBIR 第一阶段资助中，我们的目标是扩大最初的成功建立基于 PDE10A 的细胞检测方法，供业界用于开发精神分裂症疾病的创新疗法。我们将在位于马里兰州和新州的设施中进行所有研究。泽西岛拥有完成该项目所需的所有资源，我们还将聘请西弗吉尼亚大学行为医学和精神病学系的张汉廷博士，他是 PDE 研究的专家，为我们提供该项目的咨询。 SBIR 第一阶段的四个具体目标。 1. 生成稳定的 HEK293-CNG-TSHR-PDE10A 细胞系 2. 敲除 PDE4D 和 PDE4B 基因以生成。 HEK293-CNG-TSHR-PDE10A-PDE4D-/--PDE4B-/- 细胞系（PDE10A 细胞系） 3. 将 PDE10A 细胞系调整为 384 孔 HTS 格式。 4. 对化合物库进行高通量筛选，以评估鉴定已知 PDE10A 抑制剂的检测性能。第一阶段的成功完成将为将相同技术扩展到其他 PDE 亚型铺平道路，从而建立针对单个 PDE 变体的选择性细胞分析方法库，这些分析将为针对广泛治疗领域（包括 CNS、炎症、新陈代谢和癌症。