in vitro PHARMACOLOGY 2011 CATALOG - Cerep

70 in vitro pharmacology 2011 catalog
❚ serotonin
5-HT 2A - agonist radioligand
binding
Ref. 0471
Q 3 weeks
Included in:
BioPrint ® profile
Organ safety profile
Source
Ligand
Kd
Non specific
Reference
human recombinant (HEK-293 cells)
[ 125 I](±)DOI (0.1 nM)
0.3 nM
(±)DOI (1 µM)
(±)DOI (IC 50 : 0.27 nM)
Bryant, H.U. et al. (1996) Life Sci., 15: 1259-1268.
specific binding (% of control)
100
50
0
-11 -10 -9 -8 -7 -6
log [drug] (M)
(+)DOI -
serotonin
mesulergine
ketanserin
5-HT 2A
cellul ar
Ref. 3196
Ref. 3198
Q 3 weeks
Included in:
Agonist effect
Antagonist effect
Cellular functional GPCR profile
Source
human recombinant (HEK-293 cells)
Measured product IP 1
Detection method HTRF
Agonist effect Control serotonin (10 µM)
Reference serotonin (EC 50 : 27 nM)
Antagonist effect Stimulant serotonin (100 nM)
Reference ketanserin (IC 50 : 24.2 nM)
Porter, R.H.P. et al.(1999) Brit. J. Pharmacol., 128: 13-20.
Ca 2+ mobilization (% of control)
100
50
0
-12 -11 -10 -9 -8 -7 -6 -5 -4
log [agonist] (M)
serotonin
(±)DOI
5-methoxytryptamine
BW 723C86
100
50
0
-12 -11 -10 -9 -8 -7 -6 -5 -4
log [antagonist] (M)
ketanserin
mesulergine
SB 206553
SB 204741
5-HT 2A
tissue
Ref. 0334
Q 4 weeks
Source
rabbit aorta (endothelium-denuded)
Agonist serotonin (pD 2 = 6.5)
Antagonist ketanserin (pA 2 = 8.7)
Test concentrations 3 concentrations, n=2 (2 tissues)
for both activities
[Solvent] must be kept ≤ 0.1%
Rinaldi-Carmona, M. et al. (1992) J. Pharmacol. Exp. Ther., 262: 759-768.
tension (% of max.)
-12 -11
100
50
-10 -9 -8 -7 -6 -5
-9 -8 -7 -6 -5
-10 -4
0
-8 -7 -6 -5 -4 -3
log [agonist] (M)
ketanserin
none
3 nM
10 nM
30 nM
5-HT 2B - antagonist radioligand
Source
human recombinant (CHO cells)
Ligand
[ 3 H]mesulergine (2 nM)
Kd
2.4 nM
Non specific mesulergine (10 µM)
binding
Reference mesulergine (IC 50 : 2.35 nM)
Ref. 1609
Q 3 weeks
Kursar, J.D. et al. (1994) Mol. Pharmacol., 46: 227-234.
specific binding (% of control)
100
50
0
-11
-10 -9 -8 -7 -6 -5 -4
log [drug] (M)
mesulergine
SB 206553
SB 204741
ketanserin
selected cerep assays
❚ binding assays
-10 -9 -8 -7 -6 -5 -4 -3
-11
-10 -9 -8 -7 -6 -5 -4 -3
-12 -11 -10 -9 -8 -7 -6 -5 -4
Binding assays are developed according to the competition assay principle. There are three assay formats -13 -12 used: -11 -10filtration, -9 -8 -7 -6scintillation
-5
-10 -9 -8 -7 -6
proximity assay (SPA) and centrifugation. All formats utilize a radiolabeled ligand and a source of receptor (membranes, soluble/purified).
Large batches of cells, from which prepared membranes are frozen and stored, are produced in-house. Membrane preparations are
-10 -9 -8 -7 -6 -5 -4
quality controlled before use.
-12 -11 -10 -9 -8 -7 -6 -5 -4 -3
❚ cellular Functional GPCR assays
-12 -11 -10 -9 -8 -7 -6 -5
Functional GPCR assays are cell-based assays to measure agonist-like, inverse agonist, antagonist and modulator
-11 -10 -9
activity
-8 -7 -6of -5compounds.
-4
Various technologies are used: TR-FRET to determine cAMP concentration and IP1 levels, real time fluorescence to monitor calcium flux,
cellular dielectric spectroscopy to measure impedance modulation. Large batches of cells, from which whole cells are frozen and stored
for functional assays, are produced in-house. Cells are quality controlled before use. A novel patented host cell line was designed and
constructed for Gi protein coupled receptors.
❚ tissue bioassays
Tissue bioassays are functional assays designed to evaluate the agonist and antagonist activities of compounds at various receptors and
ion channels in whole tissues. The tissues used are isolated from contractile cardiac and smooth muscles of the cardiovascular, respiratory,
gastrointestinal and urogenital tracts of rodents, rabbit or human. The selected tissues are assayed in organ bath systems to record the
contractile activity in conditions as selective as possible to avoid any potential interaction with other receptors known to be present in the
tissue used.