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

描述（由申请人提供）：精神分裂症是一种慢性和毁灭性的精神障碍，影响了0.5- 0.8%的总人口和超过200万的美国人。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，导致高停药率和复发率。磷酸二酯酶10A （PDE10A）正成为治疗精神分裂症的一个新的药物靶点，有望开发出疗效更好、副作用更小的治疗方法。为了开发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分析和基于结构的药物设计。尽管基于细胞的PDE10A高通量筛选（HTS）分析将大大加快药物发现过程，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，但目前还没有商业产品可用于制药行业。Codex Biosolutions拥有专有的cAMP生物传感器技术（ACTOne”），该技术已成功开发成一种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SBIR I期融资中，我们的目标是扩大初步成功，建立基于PDE10A细胞的检测方法，该方法将被业界用于开发精神分裂症疾病的创新治疗方法。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的研究，这些设施配备了完成项目所需的所有资源。我们还将邀请到西弗吉尼亚大学行为医学与精神病学研究专家张汉廷博士为我们的项目提供咨询。我们提出SBIR第一阶段的四个具体目标。生成稳定的HEK293-CNG-TSHR-PDE10A细胞系。2. 敲除PDE4D和PDE4B基因，生成hek293 - cng - tshra -PDE10A-PDE4D-/—PDE4B-/-细胞系（PDE10A细胞系）。3. 使PDE10A细胞系适应384孔HTS格式。4. 对化合物文库进行高通量筛选，以评估鉴定已知PDE10A抑制剂的测定性能。I期的成功完成将为将相同的技术扩展到其他PDE亚型铺平道路，从而建立针对单个PDE变体的选择性细胞检测库，这将为药物发现项目提供广泛的治疗领域，包括中枢神经系统、炎症、代谢和癌症。

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.