in vitro PHARMACOLOGY 2011 CATALOG - Cerep

15
Adenosine ❚
A 2B - antagonist radioligand
binding
Ref. 0005
Q 3 weeks
Included in:
BioPrint ® profile
Organ safety profile
Source
Ligand
Kd
Non specific
Reference
human recombinant (HEK-293 cells)
[ 3 H]MRS 1754 (2 nM)
1 nM
NECA (100 µM)
NECA (IC 50 : 1.5 µM)
Ji, X. et al.(2001) Biochem. Pharmacol., 61: 657-663.
Cerep
services
Receptors
[GPCRs]
A 2B
cellul ar
Ref. 0452
Ref. 0455
Q 3 weeks
Included in:
Agonist effect
Antagonist effect
Cellular functional GPCR profile
Source
human recombinant (HEK-293 cells)
Measured product cAMP
Detection method HTRF
Agonist effect Control NECA (10 µM)
Reference NECA (EC 50 : 330 nM)
Antagonist effect Stimulant NECA (1 µM)
Reference XAC (IC 50 : 51 nM)
Cooper, J. et al. (1997) Brit. J. Pharmacol., 122: 546-550.
cAMP modulation (% of control)
100
100
50
50
0
0
-9 -8 -7 -6 -5 -4
-9 -8 -7 -6 -5 -4
log [agonist] (M)
log [antagonist] (M)
NECA
XAC
adenosine
CGS 15943
CPA
DPCPX
IB-MECA
MRS 1754
[Solvent] must be kept ≤ 0.3%
Ion
channels
Transporters
A 2B
tissue
Source
guinea-pig aorta
(precontracted with 1 µM phenylephrine)
Agonist NECA (pD 2 = 6.4)
Antagonist 8-phenyltheophylline (pA 2 = 6.2)
Test concentrations 3 concentrations, n=2 (2 tissues)
for both activities
tension (% of control)
100
50
0
8-phenyltheophylline
none
1 µM
3 µM
10 µM
Kinases
Ref. 0783
Q 4 weeks
[Solvent] must be kept ≤ 0.1%
Gurden, M.F. et al. (1993) Brit. J. Pharmacol., 109: 693-698.
-8 -7 -6 -5 -4
log [agonist] (M)
Epigenetic &
DNA-related
enzymes
A 3 - agonist radioligand
binding
Ref. 0006
Q 3 weeks
Included in:
ExpresS Profile
High-throughput profile
Diversity profile
BioPrint ® profile
Organ safety profile
Source
Ligand
Kd
Non specific
Reference
human recombinant (HEK-293 cells)
[ 125 I]AB-MECA (0.15 nM)
0.22 nM
IB-MECA (1 µM)
IB-MECA (IC 50 : 0,27 nM)
Salvatore, C.A. et al. (1993) Proc. Natl. Acad. Sci. USA, 90: 10365-10369.
Other
enzymes
Specialized
cellular
assays
A 3
cellul ar
Ref. 0982
Ref. 0983
Q 3 weeks
Included in:
Agonist effect
Antagonist effect
Cellular functional GPCR profile
Source
human recombinant (CHO cells)
Measured product cAMP
Detection method HTRF
Agonist effect Control IB-MECA (100 nM)
Reference IB-MECA (EC 50 : 1.5 nM)
Antagonist effect Stimulant IB-MECA (30 nM)
Reference MRS 1220 (IC 50 : 125 nM)
Yates, L. et al. (2006) Auton. Autacoid. Pharmacol., 26: 191-200.
[Solvent] must be kept 0.3%
❚ See other Adenosine assays, page 103
Standard
profiles
Testing
conditions
❚ For radioligand binding assays, how should I choose between the agonist and the antagonist models when both are
available?
For some binding assays two models are available using either agonist or antagonist as radioligand.
G-protein-coupled receptors have both high-affinity and low-affinity states that are bound differently by agonists and antagonists. Whereas
the antagonists bind with an equal affinity to both affinity states, agonists bind poorly to the low affinity state of the receptor. Therefore,
it is advisable to use an antagonist radioligand to evaluate the binding of antagonists knowing that this may fail to reveal the binding
of agonists. On the other hand, an assay using an agonist radioligand is suitable to evaluate both agonists and antagonists.
The testing of a compound in both assays and the comparison of its competition curves against each radioligand may provide information
about its functional activity at the receptor.
Ordering
information
Assay list
& index