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Patent Agency Ranking
公开(公告)号：KR101052621B1
公开(公告)日：2011-07-29
申请号：KR1020080095258
申请日：2008-09-29
Applicant: 한국과학기술연구원
Abstract: 본 발명은 세로토닌 6 수용체 리간드의 고효율 검색방법에 관한 것으로, HA-5-HT
6 R을 안정적으로 발현하는 세포주를 이용한 세로토닌 6 수용체 리간드의 고효율 검색방법에 관한 것이다. 본 발명의 세포주는 HA-5-HT
6 R을 안정적으로 발현함으로써, 5-HT
6 R에 선택적으로 작용하는 리간드의 검색 효율을 증가시키며, 5-HT
6 R과 결합하는 단백질 연구에 유용하게 이용될 수 있다. 이에, 5-HT
6 R이 관련된 우울증과 알츠하이머 등의 뇌질환 및 정신질환 예방과 진단 및 치료제 개발에 유용할 것이다.
세로토닌 6 수용체, HA, 리간드, 우울증, 알츠하이머Abstract translation: 本发明涉及高通量检测血清素6受体配体的方法，其中HA-5-HT
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公开(公告)号：KR1020100096501A
公开(公告)日：2010-09-02
申请号：KR1020090015404
申请日：2009-02-24
Applicant: 한국과학기술연구원
Abstract: PURPOSE: A transformed cell line which expresses human serotonin 6 receptor(5-HT6R) is provided to stably express Myc-5-HT6R on the cell surface and to use in screening human serotonin 6-receptor-specific ligand by fluorescence imaging. CONSTITUTION: A HELA cell line stably expresses Myc-5-HT6R by transducing vector containing Myc-labeled 5-HT6R. The Myc-labeld 5-HT6R gene is a gene of sequence number 1. A HELA cell line is Hela cell(deposit number KCLRF=BR-00200) which stably expresses Myc-5-HT6R. A method for screening 5-HT6R ligand of high efficiency comprises: a step of treating test chemical to a cell line; a step of measuring 5-HT6R activity of the cell line; and a step of selecting test chemical which increases or decreases 5-HT6R in the cell line.
Abstract translation: 目的：提供表达人血清素6受体（5-HT6R）的转化细胞系，以稳定表达细胞表面的Myc-5-HT6R，并用于通过荧光成像筛选人5-羟色胺6受体特异性配体。 构成：HELA细胞系通过转导含有Myc标记的5-HT6R的载体稳定表达Myc-5-HT6R。 Myc-labeld 5-HT6R基因是序列号1的基因.HELA细胞系是稳定表达Myc-5-HT6R的Hela细胞（保藏号KCLRF = BR-00200）。 用于筛选高效率的5-HT6R配体的方法包括：将化学试剂处理至细胞系的步骤; 测量细胞系的5-HT6R活性的步骤; 以及选择在细胞系中增加或减少5-HT6R的测试化学品的步骤。
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公开(公告)号：KR1020080004994A
公开(公告)日：2008-01-10
申请号：KR1020060064037
申请日：2006-07-07
Applicant: 한국과학기술연구원
IPC: G01N33/53 , G01N33/68 , G01N33/533
Abstract: A high-throughput screening(HTS) method for a 5-HT6 receptor ligand s provided to develop a selective controlling material of the 5-HT6 receptor and shorten a target protein identifying period by overcoming the defects of a conventional radio isotope technique or a non-reproducible HTS technique, thereby capable of finding an accurate mechanism of the 5-HT6 receptor and being very useful for preventing and treating brain diseases such as melancholia, cognitive impairment, and Alzheimer's disease. A high-throughput screening method for an active agent of a 5-HT6 receptor such as 5-HT, 5-CT, methoxytryptamine and tryptamine comprises the steps of: (a) transfecting the 5-HT6 receptor and a chimeric G-protein and then selecting a cell line such as HEK293, COS7, CHO-K1 and CHO-G5A and a G-protein for searching an optimum ligand through condition comparison tests; (b) introducing the selected cell line into a 96-well plate under the condition of transfecting the 5-HT6 receptor and the G-protein into DMEM(Dulbecco's modified Eagle's medium) and labeling it with a tracer dye; (c) after preparing a 96-well plate including a 5-HT6 receptor active agent with a concentration capable of activating the 5-HT6 receptor and a 96-well plate only including buffer, obtaining a Z'(signal window) value using a Ca^2+ signal regarding each of the 96-well plate including the 5-HT6 receptor active agent and the 96-well plate including buffer and determining the screening efficiency regarding the 5-HT6 receptor; and (d) after preparing 96-well plates including a 5-HT6 receptor active agent with different concentrations for activating the 5-HT6 receptor and obtaining a Ca^2+ signal in accordance with the 5-HT6 receptor activity through a fluorescent imaging technique, drawing a graph about the reaction regarding the active agent and obtaining an EC50 value of a testing material from the graph to search the active agent of the 5-HT6 receptor.
Abstract translation: 提供5-HT6受体配体的高通量筛选（HTS）方法，用于开发5-HT6受体的选择性控制材料，并通过克服常规放射同位素技术或非 - 从而能够找到5-HT6受体的精确机制，对于预防和治疗脑疾病如忧郁症，认知障碍和阿尔茨海默氏病非常有用。 用于5-HT 6受体如5-HT，5-CT，甲氧基色胺和色胺的活性剂的高通量筛选方法包括以下步骤：（a）转染5-HT 6受体和嵌合G蛋白， 然后选择细胞系如HEK293，COS7，CHO-K1和CHO-G5A和G蛋白，通过条件比较检验来搜索最佳配体; （b）在将5-HT6受体和G蛋白转染到DMEM（Dulbecco's改良的Eagle's培养基）中并用示踪剂染料标记的条件下将所选择的细胞系导入96孔板中; （c）在制备包含能够活化5-HT 6受体的浓度的5-HT 6受体活性剂和仅包含缓冲液的96孔板的96孔板后，使用 关于包括5-HT 6受体活性剂的96孔板和包含缓冲液的96孔板的每个96孔板的Ca 2+信号，并确定关于5-HT6受体的筛选效率; 和（d）在制备96孔板后，包括具有不同浓度的5-HT 6受体活性剂，用于激活5-HT 6受体，并通过荧光成像技术获得根据5-HT6受体活性的Ca 2+信号 绘制有关活性剂的反应图，并从图中获得试验材料的EC50值，以搜索5-HT6受体的活性剂。
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公开(公告)号：KR1020100035883A
公开(公告)日：2010-04-07
申请号：KR1020080095258
申请日：2008-09-29
Applicant: 한국과학기술연구원
Abstract: PURPOSE: A cell line expressing HA-5-HT6R and a method for detecting serotonin 6 receptor ligand are provided to enhance detection efficiency and to diagnose and treat brain diseases such as depression and Alzheimer. CONSTITUTION: A cell line which stably expresses HA-5-HT6R is prepared by transducing a vector containing a 5-HT6R gene construct connected with HA(hemagglutinin) to host cells. The cell line is deposited in a deposit number KCLRF-BP-00187. The host cells are HEK293, COS7, CHO-K1 or CHO-G5A. A method for detecting 5-HT6R ligand at high efficiency comprises: a step of treating a test compound to cell line which stably expressing HA-5-HT6R; a step of measuring 5-HT6R activity of treated cell line; and a step of selecting test compound which increases or decreases 5-HT6R activity in the cell line comparing to untreated cell line.
Abstract translation: 目的：提供表达HA-5-HT6R的细胞系和检测血清素6受体配体的方法，以提高检测效率，诊断和治疗脑疾病如抑郁症和阿尔茨海默病。 构成：通过将含有与HA（血凝素）连接的5-HT6R基因构建体的载体转导到宿主细胞来制备稳定表达HA-5-HT6R的细胞系。 细胞系以保藏号KCLRF-BP-00187沉积。 宿主细胞是HEK293，COS7，CHO-K1或CHO-G5A。 一种以高效率检测5-HT6R配体的方法包括：将测试化合物处理稳定表达HA-5-HT6R的细胞系的步骤; 测定处理细胞系5-HT6R活性的步骤; 以及与未处理的细胞系相比，选择增加或减少细胞系中5-HT6R活性的试验化合物的步骤。
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公开(公告)号：KR100852284B1
公开(公告)日：2008-08-14
申请号：KR1020060064037
申请日：2006-07-07
Applicant: 한국과학기술연구원
IPC: G01N33/53 , G01N33/68 , G01N33/533
Abstract: 본 발명은 형광 이미징법을 이용한 5-HT
6 수용체 리간드 고효율 검색방법에 관한 것으로서, 더욱 상세하게는 기존의 방사선 동위원소법이나 재현성이 없는 형광이미징법의 5-HT
6 수용체 리간드 검색방법의 단점을 극복하여 96-웰 플레이트 형태로 형광 이미징법을 이용한 5-HT
6 수용체 리간드 고효율 검색법(high-throughput screening, HTS)과, YTH(yeast two-hybrid) 검색법을 이용한 5-HT
6 수용체에 결합하는 Fyn 단백질 발견을 통한 Fyn 단백질이 5-HT
6 수용체의 기능을 증가시킴을 HTS의 활용법에 관한 것이다. 따라서, 본 발명은 우울증, 인지 기능, 알츠하이머 병 등과 관련된 뇌질환의 예방 및 치료에 유용하다.
5-HT6 수용체, 형광 이미징법, 고효율 검색법, YTH(yeast two-hybrid) 검색법, Fyn 단백질
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