公开(公告)号：US10584159B2

公开(公告)日：2020-03-10

申请号：US15512267

申请日：2015-09-21

申请人： The Royal Institution for the Advancement of Learning/McGill University ,  Université de Montréal

发明人： Stéphane Alain Laporte ,  Yoon Namkung ,  Michel Bouvier ,  Christian Le Gouill ,  Mireille Hogue ,  Viktoriya Lukasheva ,  Hiroyuki Kobayashi ,  Denis Deblois ,  Étienne Durette

IPC分类号： C07K14/72 ,  G01N33/50

摘要： Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use a Renilla GFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intracellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.

公开(公告)号：US20170313762A1

公开(公告)日：2017-11-02

申请号：US15512267

申请日：2015-09-21

申请人： The Royal Institution for the Advancement of Learning/McGill University ,  Université de Montréal

发明人： Stéphane Alain Laporte ,  Yoon Namkung ,  Michel Bouvier ,  Christian Le Gouill ,  Mirelle Hogue ,  Viktoriya Lukasheva ,  Hiroyuki Kobayashi ,  Denis Deblois ,  Étienne Durette

IPC分类号： C07K14/72 ,  G01N33/50

摘要： Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use a Renilla GFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intra-cellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.

公开(公告)号：US20200157184A1

公开(公告)日：2020-05-21

申请号：US16777504

申请日：2020-01-30

申请人： The Royal Institution for the Advancement of Learning/McGill University ,  Universite de Montreal

发明人： Stéphane Alain Laporte ,  Yoon Namkung ,  Michel Bouvier ,  Christian Le Gouill ,  Mireille Hogue ,  Viktoriya Lukasheva ,  Hiroyuki Kobayashi ,  Denis Deblois ,  Étienne Durette

IPC分类号： C07K14/72 ,  G01N33/50

摘要： Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use a Renilla GFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intracellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.

公开(公告)号：US20200256869A1

公开(公告)日：2020-08-13

申请号：US16755976

申请日：2018-10-15

申请人： UNIVERSITÉ DE MONTRÉAL ,  DOMAIN THERAPEUTICS

发明人： Christian Le Gouill ,  Michel Bouvier ,  Viktoriya Lukasheva ,  Mireille Hogue ,  Billy Breton

IPC分类号： G01N33/566 ,  G01N33/542

摘要： Systems and methods for the monitoring of G protein activation at various cell compartments, such as the plasma membrane and the endosomes, and in a Gα protein subunit family-selective manner are described. These systems and methods also allows the monitoring of G protein-coupled receptor (GPCR)-mediated as well as non-receptor guanine nucleotide exchange factor (GEF)-mediated G protein activation, and are based on the use of the G protein-binding domains of specific effectors of G proteins and cellular compartment markers, tagged with suitable energy (BRET) donors and acceptors.

公开(公告)号：US20170234870A1

公开(公告)日：2017-08-17

申请号：US15518888

申请日：2015-10-14

申请人： UNIVERSITÉ DE MONTRÉAL

发明人： Michel Bouvier ,  Christian Le Gouill ,  Mireille Hogue ,  Viktoriya Lukasheva

IPC分类号： G01N33/566 ,  G01N21/64 ,  C07K14/47 ,  G01N33/542 ,  C12Q1/66

CPC分类号： G01N33/566 ,  C07K14/4722 ,  C07K14/723 ,  C07K2319/00 ,  C07K2319/01 ,  C07K2319/60 ,  C12N9/0069 ,  C12Q1/66 ,  C12Y113/12007 ,  G01N21/6428 ,  G01N33/542 ,  G01N2021/6439 ,  G01N2333/4719 ,  G01N2333/726 ,  G01N2500/02

摘要： Resonance energy transfer (RET)- or protein-fragment complement assay (PCA)-based biosensors useful for assessing the activity of G-proteins are described. These biosensors are based on the competition between the Got subunit and a Gβγ interacting protein (βγ IP) for the binding to the Gβγ dimer. These biosensors comprises (1) a βγ IP and (2) a Gβ or Gγ protein; a GPCR; or a plasma membrane targeting domain, fused to suitable RET or PCA tags. Methods using such biosensors for different applications, including the identification of agents that modulates G-protein activity or for the characterization of GPCR signaling/regulation, such as G-protein preferences and activation profiles of GPCRs, are also described.

公开(公告)号：US10324085B2

公开(公告)日：2019-06-18

申请号：US15107266

申请日：2014-12-23

申请人： UNIVERSITÉ DE MONTRÉAL

发明人： David Cotnoir-White ,  Sylvie Mader ,  Étienne Gagnon ,  Michel Bouvier

IPC分类号： G01N33/68 ,  G01N33/542 ,  G01N33/74

摘要： Described herein is a bioluminescence resonance energy transfer (BRET) based system and method to monitor ternary complex formation in real-time in live cells with high sensitivity and accuracy. This system transfers energy simultaneously between a luciferase donor and intermediate and terminal acceptors, appropriately chosen to also enable transfer from the intermediate to terminal acceptor while minimizing contaminating signals. The system may also be adapted for quaternary complex detection by including a protein complementation assay (PCA) component. The system is broadly applicable to the detection of any protein ternary/quaternary complex such as those involving nuclear receptors, GPCRs, Receptor Tyrosine Kinase (RTKs), multimeric enzymes or structural proteins.

公开(公告)号：US09493456B2

公开(公告)日：2016-11-15

申请号：US14669666

申请日：2015-03-26

申请人： Université de Montréal

发明人： Michel Bouvier ,  Anne Marinier ,  Réjean Ruel ,  Patricia René ,  Yves Chantigny ,  Philippe Dagneau ,  Stéphane Gingras

IPC分类号： C07D487/00 ,  C07D471/04 ,  C07D487/04 ,  G01N33/74 ,  G01N33/50

CPC分类号： C07D471/04 ,  C07D487/04 ,  G01N33/5008 ,  G01N33/74 ,  G01N2333/72 ,  G01N2500/04 ,  G01N2500/10

摘要： Disclosed herein is a compound of Formula I: wherein X, R1, R2, and R3 are as defined herein, or a pharmaceutically acceptable salt thereof to allow the drug to penetrate the cell membrane; or a prodrug, or the compound is labeled with a detectable label or an affinity tag thereof. Also disclosed is a pharmaceutical composition, a method of treating a disorder mediated by melanocortin-4 receptors, and a method of treating obesity using the compounds described.

公开(公告)号：US20150197520A1

公开(公告)日：2015-07-16

申请号：US14669666

申请日：2015-03-26

申请人： Université de Montréal

发明人： Michel Bouvier ,  Anne Marinier ,  Réjean Ruel ,  Patricia René ,  Yves Chantigny ,  Philippe Dagneau ,  Stéphane Gingras

IPC分类号： C07D471/04 ,  C07D487/04

CPC分类号： C07D471/04 ,  C07D487/04 ,  G01N33/5008 ,  G01N33/74 ,  G01N2333/72 ,  G01N2500/04 ,  G01N2500/10

摘要： Disclosed herein is a compound of Formula I: wherein X, R1, R2, and R3 are as defined herein, or a pharmaceutically acceptable salt thereof to allow the drug to penetrate the cell membrane; or a prodrug, or the compound is labeled with a detectable label or an affinity tag thereof. Also disclosed is a pharmaceutical composition, a method of treating a disorder mediated by melanocortin-4 receptors, and a method of treating obesity using the compounds described.

摘要翻译： 本文公开了式I的化合物：其中X，R 1，R 2和R 3如本文所定义，或其药学上可接受的盐，以允许药物穿透细胞膜; 或前体药物，或者该化合物用可检测标记或其亲和标签进行标记。 还公开了药物组合物，治疗由黑皮质素-4受体介导的病症的方法，以及使用所述化合物治疗肥胖症的方法。