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

描述(申请人提供)：精神分裂症是一种慢性和破坏性的精神疾病，在美国影响0.5%-0.8%的总人口和超过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，并会产生严重的副作用，导致高度的治疗中断和复发。磷酸二酯酶10A(PDE10A)是一种治疗精神分裂症的新药靶点，有望开发出疗效更好、副作用更少的治疗药物。为了开发PDE10A抑制剂的流水线，制药和生物技术公司严重依赖无细胞酶分析和基于结构的药物设计。尽管PDE10A的细胞高通量筛选(HTS)分析将极大地加快药物发现过程，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商业产品可用。Codex Biosolution拥有一项专有的cAMP生物传感器技术(ActOne“)，该技术已成功开发为一种基于细胞的检测方法，用于在1536井HTS活动中筛选PDE4抑制剂。在这项SBIR第一阶段的资助中，我们的目标是扩大最初的成功，建立一种基于PDE10A细胞的测试，该测试将被业界用于开发精神分裂症疾病的创新疗法。我们将在我们位于马里兰州和新泽西州的设施中进行所有研究，这些设施配备了完成该项目所需的所有资源。我们还将邀请来自西弗吉尼亚大学行为医学和精神病学系的PDE研究专家张汉庭博士就该项目向我们提供咨询。1.建立稳定的HEK293-CNG-TSHR-PDE10A细胞系。2.敲除PDE4D和PDE4B基因，获得HEK293-CNG-TSHR-PDE10A-PDE4D-/--PDE4B-/-细胞系(PDE10A细胞系)。3.使PDE10A细胞系适应384孔高温超导格式。4.对化合物文库进行高通量筛选，以评估在鉴定已知PDE10A抑制剂方面的检测性能。第一阶段的成功完成将为将同样的技术扩展到其他PDE亚型铺平道路，从而建立一个针对单个PDE变体的选择性细胞分析库，这些库将服务于针对包括中枢神经系统、炎症、新陈代谢和癌症在内的广泛治疗领域的药物发现项目。 公共卫生相关性：精神分裂症是一种慢性和破坏性的精神疾病，在美国影响0.5%-0.8%的总人口和200多万人。磷酸二酯酶10A正在成为一种治疗该病的新药靶点，有望开发出疗效更好、副作用更少的治疗药物。Codex将利用专有的ActOne“技术建立一种基于PDE10A细胞的分析，这将加速制药和生物技术行业开发治疗精神分裂症疾病的创新疗法。

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. PUBLIC HEALTH RELEVANCE: 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. Phosphodiesterase 10A is emerging as a novel drug target for the disease that promises the development of therapeutics with improved efficacy and less side effects. Codex will utilize the proprietary ACTOne" technology to establish a PDE10A cell-based assay that will accelerate the pharmaceutical and biotech industry to develop innovative therapeutics for schizophrenia disease.