公开(公告)号：WO2002073199A1

公开(公告)日：2002-09-19

申请号：PCT/US2002/007434

申请日：2002-03-13

Applicant: DUKE UNIVERSITY ,  BARAK, Larry, S. ,  OAKLEY, Robert, H.

Inventor： BARAK, Larry, S. ,  OAKLEY, Robert, H.

IPC: G01N33/53

CPC classification number: C12N15/1089 ,  C12N2503/00 ,  G01N33/5076 ,  G01N33/566 ,  G01N2333/726 ,  G01N2500/04 ,  G01N2500/10

Abstract: Methods of detecting G protein-coupled receptor (GPCR) activity in vitro and in vivo are provided. In one embodiment, the method includes providing at least one cell that expresses a GPCR and a plurality of conjugated proteins. Each of the plurality of conjugated proteins is formed by conjugating an arrestin protein and a detectable molecule. The plurality of conjugated proteins are substantially evenly distributed in the cytoplasm of the at least one cell. A first image of the at least one cell is obtained by detecting an amount of energy emitted from the detectable molecules and storing a value relative to the amount of energy. The at least one cell is treated with an agonist. A second image of the at least one cell is obtained. The first image and the second image are compared to detect the localization of at least some of the plurality of conjugated proteins at endocytic vesicles and/or endosomes.

Abstract translation: 提供了在体外和/或体内检测G蛋白偶联受体（GPCR）活性的方法。 在一个实施方案中，该方法包括提供至少一个表达GPCR和多个缀合蛋白的细胞。 通过将抑制蛋白和可检测分子结合来形成多个共轭蛋白质中的每一个。 多个缀合的蛋白质基本上均匀地分布在至少一个细胞的细胞质中。 通过检测从可检测分子发射的能量的量并且存储相对于能量的值来获得至少一个细胞的第一图像。 用激动剂治疗至少一个细胞。 获得至少一个单元的第二图像。 比较第一图像和第二图像以检测多个缀合蛋白质中的至少一些在胞吞小泡和/或内体上的定位。

公开(公告)号：WO2003088924A2

公开(公告)日：2003-10-30

申请号：PCT/US2003/008208

申请日：2003-03-18

Applicant: DUKE UNIVERSITY ,  ROCKMAN, Howard, A. ,  NAGA PRASAD, Sathyamangla, V. ,  LAPORTE, Stephane, A. ,  BARAK, Larry, S. ,  CARON, Marc, G.

Inventor： ROCKMAN, Howard, A. ,  NAGA PRASAD, Sathyamangla, V. ,  LAPORTE, Stephane, A. ,  BARAK, Larry, S. ,  CARON, Marc, G.

IPC: A61K

CPC classification number: A01K67/0275 ,  A01K2217/05 ,  G01N33/566 ,  G01N2333/726 ,  G01N2500/10

Abstract: The present invention relates to compounds that alter GPCR internalization and new methods for their identification. Compounds of this invention include modified phosphoinositide 3-kinase (PI3K), modified HEAT domain, modified -adrenergic receptor kinase 1 ( ARK1), as well as other peptides or small molecules that alter GPCR internalization. The present invention also includes the use of such compounds to treat GPCR-related diseases, such as cardiovascular disease, heart failure, asthma, nephrogenic diabetes insipidus, or hypertension.

Abstract translation: 本发明涉及改变GPCR内化的化合物及其鉴定方法。 本发明的化合物包括修饰的磷酸肌醇3-激酶（PI3K），修饰的HEAT结构域，修饰的 - 肾上腺素能受体激酶1（ARK1），以及改变GPCR内化的其它肽或小分子。 本发明还包括使用这些化合物治疗GPCR相关疾病，例如心血管疾病，心力衰竭，哮喘，肾源性尿崩症或高血压。

公开(公告)号：WO2003088924A3

公开(公告)日：2003-10-30

申请号：PCT/US2003/008208

申请日：2003-03-18

Applicant: DUKE UNIVERSITY ,  ROCKMAN, Howard, A. ,  NAGA PRASAD, Sathyamangla, V. ,  LAPORTE, Stephane, A. ,  BARAK, Larry, S. ,  CARON, Marc, G.

Inventor： ROCKMAN, Howard, A. ,  NAGA PRASAD, Sathyamangla, V. ,  LAPORTE, Stephane, A. ,  BARAK, Larry, S. ,  CARON, Marc, G.

IPC: C12Q1/00

Abstract: The present invention relates to compounds that alter GPCR internalization and new methods for their identification. Compounds of this invention include modified phosphoinositide 3-kinase (PI3K), modified HEAT domain, modified -adrenergic receptor kinase 1 ( ARK1), as well as other peptides or small molecules that alter GPCR internalization. The present invention also includes the use of such compounds to treat GPCR-related diseases, such as cardiovascular disease, heart failure, asthma, nephrogenic diabetes insipidus, or hypertension.

公开(公告)号：WO2002059267A2

公开(公告)日：2002-08-01

申请号：PCT/US2002/001701

申请日：2002-01-23

Applicant: DUKE UNIVERSITY

Inventor： BARAK, Larry, S. ,  OAKLEY, Robert, H. ,  CARON, Marc, G. ,  LAPORTE, Stephane, A. ,  WILBANKS, Alyson

IPC: C12N

CPC classification number: A61K31/64 ,  A01K2217/05 ,  A61K31/41 ,  A61K38/00 ,  C07K14/705 ,  C07K14/723

Abstract: The present invention relates to modified G-protein coupled receptors (GPCRs). The modified GPCRs of the present invention include GPCRs that have been modified to have altered DRY motifs such that the modified GPCRs are constitutively desensitized. As such, the modified GPCRs of the present invention preferably localize to endocytic vesicles or endosomes in an agonist-independent manner. The invention also relates to methods of screening compounds and sample solutions for GPCR activity using the modified GPCRs.

Abstract translation: 本发明涉及修饰的G蛋白偶联受体（GPCR）。 本发明的修饰的GPCR包括已被修饰为具有改变的DRY基序的GPCR，使得修饰的GPCRs被组成型脱敏。 因此，本发明的修饰的GPCR优选以不依赖于激动剂的方式定位于内吞囊泡或内体。 本发明还涉及使用修饰的GPCR筛选化合物和GPCR活性的样品溶液的方法。