LEVISOPRENALINE L-ISOPRENALINE BDBM50407518 ISOPROTERENOL
US9492405, 27 (-)ISOPROTERENOL BDBM50407517 CHEMBL1160723
4-{1-hydroxy-2-[(propan-2-yl)amino]ethyl}benzene-1,2-diol, 2 Isoprenaline ISOPROTERONOL Isoproterenol (-) CHEMBL434 4-[1-hydroxy-2-(propan-2-ylamino)ethyl]benzene-1,2-diol Isoproterenol BDBM25392 Isoproterenol,(+) Norisodrine Isoproterenol-l Novodrin 4-[1-hydroxy-2-(isopropylamino)ethyl]pyrocatechol;hydrochloride
3,4-Dihydroxy-alpha-((isopropylamino)methyl)benzyl alcohol Diazossido (+-)-Isoprenaline BDBM50241108 Isoprenaline CHEMBL181 N-Isopropylnorepinephrine N-Isopropylnoradrenaline 4-[1-hydroxy-2-(propan-2-ylamino)ethyl]benzene-1,2-diol7-chloro-3-methyl-2H-1,2,4-benzothiadiazine 1,1-dioxide Isoproterenol DIAZOXIDE
cid_5807 4-[1-hydroxy-2-(propan-2-ylamino)ethyl]benzene-1,2-diol;hydrochloride DL-ISOPROTERENOL HYDROCHLORIDE SMR000058267 BDBM84344 MLS001335893 4-[1-hydroxy-2-(isopropylamino)ethyl]pyrocatechol;hydrochloride 4-[1-oxidanyl-2-(propan-2-ylamino)ethyl]benzene-1,2-diol;hydrochloride
cid_6852409 4-[(1S)-1-hydroxy-2-(isopropylamino)ethyl]pyrocatechol;tartaric acid 2,3-bis(oxidanyl)butanedioic acid;4-[(1S)-1-oxidanyl-2-(propan-2-ylamino)ethyl]benzene-1,2-diol SMR000058532 2,3-dihydroxybutanedioic acid;4-[(1S)-1-hydroxy-2-(propan-2-ylamino)ethyl]benzene-1,2-diol BDBM34652 (+)-ISOPROTERENOL (+)-BITARTRATE SALT MLS000028736
- Bosak, A; Gazic Smilovic, I; Sinko, G; Vinkovic, V; Kovarik, Z Metaproterenol, isoproterenol, and their bisdimethylcarbamate derivatives as human cholinesterase inhibitors. J Med Chem 55: 6716-23 (2012)
- Brink, CB; Wade, SM; Neubig, RR Agonist-directed trafficking of porcine alpha(2A)-adrenergic receptor signaling in Chinese hamster ovary cells: l-isoproterenol selectively activates G(s). J Pharmacol Exp Ther 294: 539-47 (2000)
- ChEMBL_38798 (CHEMBL652339) Evaluated for Adenylyl cyclase activation as % of the maximal stimulation with isoproterenol at 100 nM
- ChEBML_1689808 Inhibition of human GRK2 expressed in HEK-B2 cells assessed as isoproterenol-stimulated cAMP accumulation preincubation for 20 mins followed by isoproterenol stimulation measured after 20 mins by cAMP ALPHA-screen assay
- ChEMBL_1689808 (CHEMBL4040378) Inhibition of human GRK2 expressed in HEK-B2 cells assessed as isoproterenol-stimulated cAMP accumulation preincubation for 20 mins followed by isoproterenol stimulation measured after 20 mins by cAMP ALPHA-screen assay
- ChEMBL_1801542 (CHEMBL4273834) Inhibition of Gs coupled adrenergic beta2 receptor in HEK293 cells assessed as reduction in isoproterenol-induced cAMP production pretreated for 1 hr followed by isoproterenol addition and measured after 30 mins by FRET assay
- ChEMBL_1873769 (CHEMBL4375058) Antagonist activity at beta2 adrenergic receptor (unknown origin) expressed in HEK293 cell assessed as inhibition of isoproterenol-induced cAMP production preincubated for 15 mins followed by isoproterenol addition and measured after 15 mins
- ChEMBL_1903849 (CHEMBL4406071) Agonist activity at human Gi/o-coupled D3 receptor expressed in HEK293T cells assessed as inhibition of isoproterenol-induced cAMP accumulation preincubated for 15 mins followed by isoproterenol addition by Glosensor-based luminescence assay
- ChEMBL_1892552 (CHEMBL4394473) Partial agonist activity at human Gi/o-coupled D2R expressed in HEK293T cells assessed as inhibition of isoproterenol-induced cAMP accumulation preincubated for 15 mins followed by isoproterenol-stimulation and measured by Glosensor-based luminescence assay
- ChEMBL_2110690 (CHEMBL4819540) Agonist activity at beta2 adrenoceptor (unknown origin) expressed in HEK293 cells assessed as increase in isoproterenol-induced cAMP production preincubated for 20 mins followed by isoproterenol stimulation and measured after 5 mins by chemiluminescence-based assay
- ChEMBL_1742451 (CHEMBL4158201) Agonist activity at FFA2R in mouse mature adipocytes assessed as inhibition of isoproterenol-induced glycerol production
- ChEMBL_349361 (CHEMBL865067) Inhibition of isoproterenol-induced increase of cAMP in HEK293 cell expressing human adenosine A3 receptor
- ChEMBL_1873766 (CHEMBL4375055) Antagonist activity at beta1 adrenergic receptor (unknown origin) stably expressed in HEK293 cell membranes assessed as inhibition of isoproterenol-induced cAMP production preincubated for 15 mins followed by isoproterenol addition and measured after 30 mins by TR-FRET assay
- ChEMBL_1873767 (CHEMBL4375056) Antagonist activity at beta2 adrenergic receptor (unknown origin) stably expressed in HEK293 cell membranes assessed as inhibition of isoproterenol-induced cAMP production preincubated for 15 mins followed by isoproterenol addition and measured after 30 mins by TR-FRET assay
- ChEMBL_31398 (CHEMBL645644) Inhibition of isoproterenol-stimulated cAMP accumulation in HEK 293 cells expressing human Adenosine A3 receptor
- ChEMBL_1774534 (CHEMBL4231526) Antagonist activity at human beta3 adrenergic receptor expressed in HEK293 cells co-expressing Galpha16 assessed as inhibition of isoproterenol-induced intercellular calcium mobilization pre-incubated for 10 mins before isoproterenol addition by Fluo-4AM dye-based fluorescence assay
- ChEMBL_1774535 (CHEMBL4231527) Antagonist activity at human beta2 adrenergic receptor expressed in HEK293 cells co-expressing Galpha16 assessed as inhibition of isoproterenol-induced intercellular calcium mobilization pre-incubated for 10 mins before isoproterenol addition by Fluo-4AM dye-based fluorescence assay
- ChEMBL_1774536 (CHEMBL4231528) Antagonist activity at human beta1 adrenergic receptor expressed in HEK293 cells co-expressing Galpha16 assessed as inhibition of isoproterenol-induced intercellular calcium mobilization pre-incubated for 10 mins before isoproterenol addition by Fluo-4AM dye-based fluorescence assay
- ChEMBL_1807450 (CHEMBL4306809) Agonist activity at human Muscarinic acetylcholine receptor M4 expressed in HEK cells co-expressing Glosensor construct assessed as decrease in isoproterenol-induced cAMP accumulation incubated for 7 mins in presence of isoproterenol by Glosensor cAMP reagent/plate reader based luminescence assay
- ChEMBL_1807452 (CHEMBL4306811) Agonist activity at human Muscarinic acetylcholine receptor M2 expressed in HEK cells co-expressing Glosensor construct assessed as decrease in isoproterenol-induced cAMP accumulation incubated for 7 mins in presence of isoproterenol by Glosensor cAMP reagent/plate reader based luminescence assay
- ChEBML_29593 Adenosine receptor agonistic activity against isoproterenol (1 uM) stimulated cAMP accumulation in DDT MF-2 cells
- ChEMBL_472907 (CHEMBL922121) Agonist activity at adenosine A1 receptor assessed as inhibition of isoproterenol-stimulated cAMP accumulation in DDT1MF-2 cells
- ChEBML_37979 Beta-2 adrenergic receptor blocking was assessed from 50% inhibition of the isoproterenol submaximal response on isolated Guinea pig trachea(IT)
- ChEMBL_451676 (CHEMBL901889) Agonist activity at adenosine A1 receptor expressed in DDT1 MF2 cells assessed as inhibition of (-)-isoproterenol-stimulated cAMP accumulation
- ChEBML_38603 Compound was evaluated for its binding affinity towards Beta-2 adrenergic receptor in rat soleus membrane by displacing (-)-isoproterenol (50 uM)
- ChEBML_38604 Compound was evaluated for its binding affinity towards beta-2 adrenergic receptor in rat soleus membrane by displacing (-)-isoproterenol (50 uM)
- ChEBML_39187 Beta-1 adrenergic receptor blocking was assessed from 50% inhibition of the isoproterenol submaximal response on isolated Guinea pig atria(IA)
- ChEMBL_1539001 (CHEMBL3738004) Antagonist activity at human beta2 receptor expressed in CHO-K1 cells assessed as isoproterenol-induced cAMP level by HTRF assay
- ChEMBL_1539002 (CHEMBL3738005) Antagonist activity at human beta1 receptor expressed in CHO-K1 cells assessed as isoproterenol-induced cAMP level by HTRF assay
- ChEMBL_625100 (CHEMBL1112549) Inhibition of PDE4B3 expressed in CHO cells assessed as isoproterenol-induced [125I]cAMP accumulation after 15 mins by scintillation counting
- ChEMBL_625101 (CHEMBL1112550) Inhibition of PDE4D4 expressed in CHO cells assessed as isoproterenol-induced [125I]cAMP accumulation after 15 mins by scintillation counting
- ChEBML_216165 Tested for beta-1-adrenergic blocking effect by measuring the ability to inhibit the positive inotropic effect of isoproterenol on the isolated right atrium of guinea pigs
- ChEMBL_216165 (CHEMBL815336) Tested for beta-1-adrenergic blocking effect by measuring the ability to inhibit the positive inotropic effect of isoproterenol on the isolated right atrium of guinea pigs
- ChEMBL_39350 (CHEMBL654957) In vitro beta-1 adrenergic receptor activity was determined via inhibition of the positive chronotropic actions of isoproterenol in isolated guinea pig atrial preparations
- ChEMBL_665484 (CHEMBL1261259) Antagonist activity at human recombinant adrenergic beta2 receptor expressed in CHO cells assessed as inhibition of isoproterenol-induced cyclic AMP formation by HTRF assay
- ChEMBL_665490 (CHEMBL1261321) Antagonist activity at human recombinant adrenergic beta3 receptor expressed in CHO cells assessed as inhibition of isoproterenol-induced cyclic AMP formation by HTRF assay
- ChEMBL_859681 (CHEMBL2169116) Agonist activity at human D2L receptor expressed in HEK293T cells coexpressing Gi subunit assessed as inhibition of isoproterenol-stimulated cAMP production by luminescence assay
- ChEMBL_1793338 (CHEMBL4265257) Activation of human Y1R expressed in HEK293 cells assessed as inhibition of isoproterenol-induced increase in intracellular cAMP levels by calcium 5 dye-based FLIPR assay
- ChEMBL_1793339 (CHEMBL4265258) Activation of human Y2R expressed in HEK293 cells assessed as inhibition of isoproterenol-induced increase in intracellular cAMP levels by calcium 5 dye-based FLIPR assay
- ChEMBL_1793340 (CHEMBL4265259) Activation of human Y4R expressed in HEK293 cells assessed as inhibition of isoproterenol-induced increase in intracellular cAMP levels by calcium 5 dye-based FLIPR assay
- ChEMBL_1793341 (CHEMBL4265260) Activation of human Y5R expressed in HEK293 cells assessed as inhibition of isoproterenol-induced increase in intracellular cAMP levels by calcium 5 dye-based FLIPR assay
- ChEMBL_38001 (CHEMBL651635) In vitro beta-2 adrenergic receptor activity was determined by measuring inhibition of the isoproterenol induced relaxation in isolated guinea pig tracheal chains contracted with PGF2-alpha
- ChEMBL_578238 (CHEMBL1051553) Agonist activity at human cloned adrenergic beta3 receptor expressed in CHO cells assessed as [125I]cAMP accumulation after 2 days by radioimmunoassay relative to isoproterenol
- ChEMBL_578240 (CHEMBL1051555) Agonist activity at human cloned adrenergic beta2 receptor expressed in CHO cells assessed as [125I]cAMP accumulation after 2 days by radioimmunoassay relative to isoproterenol
- ChEMBL_859677 (CHEMBL2168914) Partial agonist activity at human D2L receptor expressed in HEK293T cells coexpressing Gi subunit assessed as inhibition of isoproterenol-stimulated cAMP production by luminescence assay
- ChEMBL_859788 (CHEMBL2169616) Agonist activity at human A1AR expressed in HEK293T/17 cells assessed as inhibition of isoproterenol-induced cAMP accumulation incubated for 10 mins by luciferase reporter assay
- ChEMBL_1329165 (CHEMBL3225181) Antagonist activity at LPAR1/LPAR3 in rat glioma C62B cells assessed as inhibition of LPA-induced reduction in isoproterenol-stimulated [3H]cAMP accumulation by liquid scintillation counting
- ChEMBL_578242 (CHEMBL1051557) Agonist activity at human cloned adrenergic beta-1 receptor expressed in CHO cells assessed as [125I]cAMP accumulation after 2 days by radioimmunoassay relative to isoproterenol
- ChEMBL_1282125 (CHEMBL3102009) Agonist activity at human NPYY5 receptor expressed in HEK293 cells assessed as decrease in isoproterenol-induced cAMP level measured every 30 secs for 360 secs by FLIPR assay
- ChEMBL_1282126 (CHEMBL3102010) Agonist activity at human NPYY4 receptor expressed in HEK293 cells assessed as decrease in isoproterenol-induced cAMP level measured every 30 secs for 360 secs by FLIPR assay
- ChEMBL_1282127 (CHEMBL3102011) Agonist activity at human NPYY2 receptor expressed in HEK293 cells assessed as decrease in isoproterenol-induced cAMP level measured every 30 secs for 360 secs by FLIPR assay
- ChEMBL_1282128 (CHEMBL3102012) Agonist activity at human NPYY1 receptor expressed in HEK293 cells assessed as decrease in isoproterenol-induced cAMP level measured every 30 secs for 360 secs by FLIPR assay
- ChEMBL_1699995 (CHEMBL4050977) Antagonist activity at human KOR expressed in HEK293T cells coexpressing Gi assessed as Gi-mediated inhibition of cAMP production incubated for 15 mins followed by Gs activator isoproterenol addition measured after 15 mins by luciferase reporter gene assay
- ChEMBL_1903851 (CHEMBL4406073) Antagonist activity at human Gi/o-coupled D3 receptor expressed in HEK293T cells assessed as suppression of dopamine-induced inhibition of isoproterenol-stimulated cAMP accumulation preincubated for 10 mins followed by dopamine stimulation and measured after 20 mins by Glosensor-based luminescence assay
- cAMP GloSensor Assay HEK293T cells co-expressing the cAMP biosensor GloSensor-22F (Promega) and hD2 receptors were seeded (10,000 cells/20 ul/well) into white, clear-bottom, tissue culture plates in HBSS, 10% FBS, 20 mM HEPES, pH 7.4. After a one- to two-hour recovery, cells were treated with 10 ul of 3x test or reference drug prepared in HBSS, 10% FBS, 20 mM HEPES, pH 7.4. After 30 minutes, cells were challenged with 10 ul of 1,200 nM (4x) isoproterenol in 8% (4x) GloSensor reagent. Luminescence per well per second was read on a Wallac TriLux microbeta plate counter. Data were normalized to the isoproterenol response (100%) and the maximal quinpirole-induced inhibition thereof (0%) and regressed using the sigmoidal dose-response function built into GraphPad Prism 4.0. Notably, HEK293T cells expressing the GloSensor-22F alone (no hD2) were assayed in parallel and displayed no inhibition of isoproterenol-stimulated cAMP.
- ChEMBL_1807451 (CHEMBL4306810) Positive allosteric modulatory activity at human Muscarinic acetylcholine receptor M4 expressed in HEK cells co-expressing Glosensor construct assessed as as increase in acteylcholine-induced cAMP accumulation incubated for 7 mins in presence of isoproterenol/acetylcholine by Glosensor cAMP reagent/plate reader based luminescence assay
- Homogeneous Time-Resolved Fluorescence Assay Compounds were dissolved and serially diluted (5-fold) in DMSO to generate a 10-point dose response stock. The stock was then diluted 100-fold in assay buffer (PBS containing 1 mM IBMX) before a volume of 2.5 μL was added to the cells (the final, top concentration of compound in the dose-response is typically 10 or 100 μM). After a brief incubation, 2.5 μL of isoproterenol stock, prepared at a concentration 4 times its EC90 at the receptor of interest, was added to the wells. The EC90 for isoproterenol, a beta-adrenergic agonist, was determined in separate experiments using standard methods to measure agonist potencies.Following a 1-hour incubation at room temperature, 5 μL of cAMP-D2 Reagent diluted in Lysis Buffer was added to each well followed by 5 μL of Cryptate Reagent. Plates were further incubated at room temperature for 1 hour prior to reading. Time resolved fluorescence measurements were collected on a suitable, HTRF-capable plate reader.
- Homogeneous Time-Resolved Fluorescence (HTRF) cAMP Antagonist Assay HTRF cAMP assays were performed according to manufacturer's instructions (Cisbio, cAMP Dynamic 2 Assay Kit; #62AM4PEJ). CHO-K1 cells stably expressing recombinant receptor were harvested and suspended in warm PBS to make a 300,000 cells/mL stock. This cell suspension was dispensed into 384 well assay plates (PerkinElmer ProxiPlate #6008280) at 5 μL per well (1500 cells/well) along with a cAMP standard curve.Compounds were dissolved and serially diluted (5-fold) in DMSO to generate a 10-point dose response stock. The stock was then diluted 100-fold in assay buffer (PBS containing 1 mM IBMX) before a volume of 2.5 μL was added to the cells (the final, top concentration of compound in the dose-response is typically 10 or 100 μM). After a brief incubation, 2.5 μL of isoproterenol stock, prepared at a concentration 4 times its EC90 at the receptor of interest, was added to the wells. The EC90 for isoproterenol, a beta-adrenergic agonist, was determined in separate experiments using standard methods to measure agonist potencies.Following a 1-hour incubation at room temperature, 5 μL of cAMP-D2 Reagent diluted in Lysis Buffer was added to each well followed by 5 μL of Cryptate Reagent. Plates were further incubated at room temperature for 1 hour prior to reading. Time resolved fluorescence measurements were collected on a suitable, HTRF-capable plate reader.Counts from the plate reader were fit to the cAMP standard curve on the assay plate in order to determine cAMP concentrations in each well, and these values were used to construct dose-response curves to obtain IC50 values.
- cAMP Inhibition Assay HEK293T cells were co-transfected with plasmids encoding the cAMP biosensor GloSensor-22F (Promega) and the different human KOR variants as described [Allen et al., Proc. Natl. Acad. Sci. U.S.A., 108:18488-18493]. After an 18-h incubation at 37 °C, the cells were seeded (at 20,000 cells/20 μl/well) into white, clear bottom, 384-well tissue culture plates precoated with poly-L-lysine (25 mg/liter; Sigma). Cells were plated in DMEM containing 1% dialyzed FBS. After a 24-h recovery, the medium was replaced with 20 μl of Drug Buffer (1× Hanks' balanced salt solution, 20 mM HEPES, pH 7.4), and the cells were treated with 10 μl of 3× test or reference drug prepared in Drug Buffer. After 20 min, cAMP production was stimulated and detected by treatment with 10 μl of 1.2 μM (4×) isoproterenol in 8% (4×) GloSensor reagent. Luminescence per well per second was read on a Wallac Micro-Beta TriLux plate scintillation counter.
- cAMP Gs dynamic HTRF assay Compound preparation. Candidate beta-adrenergic compounds, dissolved to 10 mM in DMSO, were diluted in 1× stimulation buffer 1 (Cisbio Part #64SB1FDD) containing 1 mM 3-Isobutyl-1-methylxanthene (IBMX; Cayman Chemical Company, catalog #13347). Serial dilutions were made in a 96 well V-bottom polypropylene compound microplate (Corning, catalog #3363) in stimulation buffer containing 1 mM IBMX, to 2× of the final desired concentration. Standard serial dilution curves were 10-point, 5-fold dilutions starting from a highest concentration of 10 μM. Controls present on every assay plate were 0.1% DMSO (vehicle control), 1 μM isoproterenol (full beta-adrenergic agonist control) and 15 μM xamoterol (partial beta-adrenergic agonist control). 5 μL from the 2× compound plate was stamped into a white 384 round well small volume HiBase assay plate (Greiner Bio-One; catalog #784075) to provide 4 technical replicates per concentration, per compound. Assay plates were centrifuged at 500×g for 10 seconds. Compounds and IBMX were prepared at 2× final dose to compensate for addition of cells.Cell preparation. 1× stimulation buffer, washing PBS (Dulbecco's phosphate-buffered saline, −Mg −Ca; Caisson Labs, catalog #PBL01), assay PBS (Dulbecco's phosphate-buffered saline, +Mg, +Ca; Caisson Labs, catalog #PBL02) and Versene (0.02% EDTA disodium salt solution in PBS without calcium or magnesium; Caisson Labs, catalog #EDL01) were pre-warmed to 37° C. Cells expressing beta-adrenergic receptor were washed in washing PBS to remove growth medium and then released from the surface by incubating with Versene for 5-10 minutes at 37° C. Cells were harvested using assay PBS, counted manually by hemocytometer or by an automated cell counter, pelleted by centrifugation (200×g, 5 minutes) and resuspended in 37° C. 1× stimulation buffer to a final density of 1.5×10{circumflex over ( )}6 cells/mL. 5 μL of the suspended cell solution (7500 cell total) were added to all wells of the 384 well assay plate, the assay plate was covered with an Axygen plate seal (Corning PCR-SP) and incubated in a humidified 37° C. environment supplemented with 5% CO2 for 30 minutes.HTRF reagent addition, reading and data analysis. After 30 minutes of cell stimulation with test compound, the assay plates were centrifuged at 500×g for 10 seconds, and incubation was stopped with the addition of 5 μL cAMP-D2 acceptor, diluted 1:21 in detection and lysis buffer 2 (Cisbio 62CL2FDF) was added to all cells. Subsequently, 5 μL Anti-cAMP-Eu Donor, diluted 1:21 in detection and lysis buffer 2, was added to cells. Plates were sealed and reactions gently vortexed at 900 rpm on a Heidolph Titramax 1000 for at least 30 minutes at room temperature. Plates were centrifuged again at 500×g for 10 seconds, and HTRF was measured using a Tecan Spark plate reader at 50 flashes per well. HTRF ratios (665 nm/620 nm×10,000) were determined and plotted in GraphPad Prism to generate a concentration-effect curve. Potency estimates (EC50) were derived from the four-parameter nonlinear regression of the concentration-effect curve and an estimate of relative efficacy was determined by comparing the magnitude of the test compound HTRF signal window (min−max dose) with the signal window of the full agonist control, isoproterenol.