Discovery of β2 Adrenergic Receptor Ligands Using Biosensor Fragment Screening of Tagged Wild-Type Receptor.
G-protein coupled receptors (GPCRs) are the primary target class of currently marketed drugs, accounting for about a quarter of all drug targets of approved medicines. However, almost all the screening efforts for novel ligand discovery rely exclusively on cellular systems overexpressing the receptors. An alternative ligand discovery strategy is a fragment-based drug discovery, where low molecular weight compounds, known as fragments, are screened as initial starting points for optimization. However, the screening of fragment libraries usually employs biophysical screening methods, and as such, it has not been routinely applied to membrane proteins. We present here a surface plasmon resonance biosensor approach that enables, cell-free, label-free, fragment screening that directly measures fragment interactions with wild-type GPCRs. We exemplify the method by the discovery of novel, selective, high affinity antagonists of human β2 adrenoceptor.
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Tonia
T
Aristotelous
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States.
Seungkirl
S
Ahn
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States.
Arun K
AK
Shukla
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Sylwia
S
Gawron
NIMH Psychoactive Drug Screening Program, Department of Pharmacology, The University of North Carolina Chapel Hill School of Medicine , Chapel Hill, North Carolina 27759, United States.
Maria F
MF
Sassano
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States.
Alem W
AW
Kahsai
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States.
Laura M
LM
Wingler
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States.
Xiao
X
Zhu
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States.
Prachi
P
Tripathi-Shukla
NIMH Psychoactive Drug Screening Program, Department of Pharmacology, The University of North Carolina Chapel Hill School of Medicine , Chapel Hill, North Carolina 27759, United States.
Xi-Ping
XP
Huang
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Jennifer
J
Riley
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Jérémy
J
Besnard
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Kevin D
KD
Read
NIMH Psychoactive Drug Screening Program, Department of Pharmacology, The University of North Carolina Chapel Hill School of Medicine , Chapel Hill, North Carolina 27759, United States ; Department of Pharmacology and Division of Chemical Biology and Medicinal Chemistry, The University of North Carolina Chapel Hill School of Medicine , Chapel Hill, North Carolina 27759, United States.
Bryan L
BL
Roth
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Ian H
IH
Gilbert
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Andrew L
AL
Hopkins
Department of Medicine, Duke University Medical Center , Durham, North Carolina 27710, United States ; Howard Hughes Medical Institute, Duke University Medical Center , Durham, North Carolina 27710, United States ; Department of Biochemistry, Duke University Medical Center , Durham, North Carolina 27710, United States.
Robert J
RJ
Lefkowitz
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
Iva
I
Navratilova
Fragment screening; G-protein coupled receptors; surface plasmon resonance; β2 adrenoceptor