| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg | |||
| 250mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
Endothelin ETA receptor (IC50 = 360 ± 160 nM on recombinant human ETA; IC50 = 220 ± 60 nM on human smooth muscle cells; IC50 = 430 ± 140 nM on rat mesangial cells)
Endothelin ETB receptor (IC50 = 530 ± 150 nM on recombinant human ETB; IC50 = 160 ± 77 nM on human placenta membranes; IC50 = 227 ± 92 nM on porcine cerebellum membranes; IC50 = 1,140 ± 340 nM on rat endothelial cells) |
|---|---|
| ln Vitro |
Ro 46-2005 competed for [125I]ET-1 binding on human smooth muscle cells (ETA) with IC50 = 220 ± 60 nM, and on rat mesangial cells (ETA) with IC50 = 430 ± 140 nM. [1]
On ETB-containing preparations, it competed with IC50 = 160 ± 77 nM (human placenta), 227 ± 92 nM (porcine cerebellum), and 1,140 ± 340 nM (rat endothelial cells). [1] On recombinant human ETA receptors, Ro 46-2005 showed IC50 = 360 ± 160 nM for [125I]ET-1 binding; on recombinant human ETB receptors, IC50 = 530 ± 150 nM. [1] It competed similarly for [125I]ET-1, [125I]ET-2, and [125I]ET-3 binding on recombinant ETA (IC50 = 360 ± 160 nM, 397 ± 53 nM, and 212 ± 45 nM, respectively) and on human placenta ETB (IC50 = 160 ± 77 nM, 97 ± 5 nM, and 113 ± 4 nM, respectively). [1] Saturation binding experiments showed that Ro 46-2005 increased the apparent KD for ET-1 without changing Bmax, indicating competitive binding on both ETA and ETB receptors. On recombinant ETA receptors, KD increased from 68 pM (control) to 163 pM (0.65 µM Ro 46-2005), 198 pM (1.3 µM), and 477 pM (2.5 µM), with constant Bmax ~3,000 fmol/mg. On placental ETB receptors, KD increased from 20 pM (control) to 29 pM (0.1 µM), 57 pM (0.2 µM), and 82 pM (0.4 µM), with constant Bmax ~138 fmol/mg. [1] Ro 46-2005 inhibited ET-1-induced arachidonic acid release from rat mesangial cells with an IC50 of 1.8 ± 0.3 µM. [1] |
| ln Vivo |
In rats, Ro 46-2005 was shown to prevent cerebral vasoconstriction following subarachnoid hemorrhage and post-ischemic renal vasoconstriction (as referenced in discussion, but no detailed experimental data provided in this paper). [1]
|
| Enzyme Assay |
Competition binding assays were performed on membrane preparations (human placenta, porcine cerebellum, baculovirus-infected insect cells expressing recombinant ETA or ETB receptors) or on whole attached cells (rat mesangial cells, human smooth muscle cells, rat endothelial cells). Membrane assays were conducted in 250 µl of 50 mM Tris buffer (pH 7.4) containing 25 mM MnCl2, 1 mM EDTA, 0.5% (w/v) BSA, with 5-35 µg protein, 32 pM [125I]-labeled ET (or 213 pM [125I]ET-3 for ETA), and increasing concentrations of unlabeled ligands. After 2 h incubation at 22°C, bound and free ligand were separated by filtration. Non-specific binding was assessed with 100 nM unlabeled ET-1. IC50 values were determined after logit/log transformation. [1]
Saturation binding experiments were performed with membranes of recombinant ETA receptor (5 µg protein/well) or human placenta membranes (35 µg protein/well, ~90% ETB). Membranes were incubated for 3 h with increasing [125I]ET-1 (6.4-660 pM) in the presence of varying concentrations of Ro 46-2005. Non-specific binding was determined with 100 nM unlabeled ET-1. KD and Bmax were calculated by linear regression analysis of Scatchard plots. [1] For arachidonic acid release measurement, confluent rat mesangial cells in 24-well plates (400,000 cells/well) were incubated for 24 h with [3H]arachidonic acid (1 µCi/well), washed three times with DMEM containing 2 mg/ml BSA and 25 mM HEPES (pH 7.4), then incubated in this medium for 30 min at 37°C. After an additional wash, the reaction was started by adding fresh medium containing various amounts of ET-1 and Ro 46-2005. All steps were performed with or without 100 µM phosphoramidon. After 0-30 min incubation, supernatant was removed, centrifuged to sediment detached cells, and [3H]arachidonic acid release was quantified by liquid scintillation counting. [1] |
| Cell Assay |
Binding assays on whole attached cells: Rat mesangial cells, human smooth muscle cells, or rat endothelial cells were cultured in 24-well plates. Cells were incubated in 500 µl DMEM containing 2 mg/ml BSA and 25 mM HEPES with 35 pM [125I]ET-1 and increasing concentrations of Ro 46-2005 for 2 h at 22°C. After extensive washing, cells were solubilized in 1% (w/v) SDS, 0.5 M NaOH, 100 mM EDTA. Non-specific binding was assessed with 100 nM unlabeled ET-1. Each assay was performed three times in triplicate. [1]
Arachidonic acid release assay (functional cell assay): Rat mesangial cells were pre-labeled with [3H]arachidonic acid as described in Enzyme Assay. ET-1 induced a dose-dependent release with an EC50 of 4.4 nM and maximal stimulation of 870 ± 40% above baseline at 330 nM ET-1. In the presence of 0.1 mM phosphoramidon, the maximal response increased to 1,400 ± 60% without affecting EC50. Ro 46-2005 (0.1-100 µM) dose-dependently inhibited the ET-1 (10 nM)-stimulated arachidonic acid release measured after 30 min incubation in the presence of 0.1 mM phosphoramidon, with an IC50 of 1.8 ± 0.3 µM. [1] |
| Animal Protocol |
Renal ischaemia model: Male Wistar rats were anaesthetized and instrumented. Renal ischaemia was induced by clamping the left renal artery at its origin for 45 min. Ten min before ischaemia, rats received an intravenous bolus injection of Ro 46-2005 (3 mg/kg, n=6) or saline vehicle (n=7). Upon reperfusion, renal blood flow stabilized within 20-30 min, and percent decreases were measured [2].
Subarachnoid haemorrhage (SAH) model: Rats were prepared and SAH induced as described. Ro 46-2005 (3 mg/kg, n=9) or glucose placebo (n=10) was injected intravenously 10 min before SAH in a blinded, randomized fashion. Cerebral blood flow (CBF) was measured before and 30, 60, 120 min after SAH using the radioactive microspheres technique. Percent changes in cerebellar blood flow were calculated [2]. Squirrel monkey blood pressure model: Sodium depletion was achieved by intramuscular injection of furosemide (3 mg/kg) for three consecutive days before the experiment. Ro 46-2005 (10, 30, 100 mg/kg, n=5,5,4 respectively) or control vehicle (n=6) was administered by gavage. Blood pressure was monitored in conscious, unrestrained monkeys chronically instrumented with a telemetry system. Baseline mean arterial pressure was 96-99 mmHg. Change in mean arterial pressure was recorded after drug administration [2]. |
| References |
|
| Additional Infomation |
Endothelin (ET) is a potent vasoconstrictor peptide with three isopeptides (ET-1, ET-2, ET-3). Two G-protein-coupled ET receptor subtypes exist: ETA (selective for ET-1/ET-2 over ET-3) and ETB (binds all three with similar potency). ETA receptors on smooth muscle mediate vasoconstriction; ETB receptors on endothelium mediate vasodilation, but vasoconstrictive ETB receptors have also been described. The peptide ETA-selective antagonists BQ-123 and FR-139317 potently blocked recombinant ETA (IC50 = 63 ± 16 nM and 13 ± 4 nM, respectively) but did not compete for ETB binding up to 100 µM. The ETB-selective ligand sarafotoxin S6c did not bind to ETA but had IC50 = 0.120 ± 0.01 nM on recombinant ETB. Ro 46-2005 is a non-peptide antagonist that blocks both ETA and ETB competitively, which may be advantageous in diseases where both subtypes contribute to vasoconstriction. It has been shown to prevent cerebral vasoconstriction after subarachnoid hemorrhage and post-ischemic renal vasoconstriction in rats. [1]
|
| Molecular Formula |
C23H27N3O6S
|
|---|---|
| Molecular Weight |
473.544
|
| Exact Mass |
473.162
|
| Elemental Analysis |
C, 58.34; H, 5.75; N, 8.87; O, 20.27; S, 6.77
|
| CAS # |
150725-87-4
|
| Related CAS # |
150725-87-4
|
| PubChem CID |
122044
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
617.5±65.0 °C at 760 mmHg
|
| Flash Point |
327.3±34.3 °C
|
| Vapour Pressure |
0.0±1.9 mmHg at 25°C
|
| Index of Refraction |
1.593
|
| LogP |
2.22
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
33
|
| Complexity |
687
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=S(C1=CC=C(C(C)(C)C)C=C1)(NC2=NC=NC(OCCO)=C2OC3=CC=CC(OC)=C3)=O
|
| InChi Key |
ZNXOKLWCOWOECF-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C23H27N3O6S/c1-23(2,3)16-8-10-19(11-9-16)33(28,29)26-21-20(22(25-15-24-21)31-13-12-27)32-18-7-5-6-17(14-18)30-4/h5-11,14-15,27H,12-13H2,1-4H3,(H,24,25,26)
|
| Chemical Name |
4-tert-butyl-N-[6-(2-hydroxyethoxy)-5-(3-methoxyphenoxy)pyrimidin-4-yl]benzenesulfonamide
|
| Synonyms |
RO462005; RO-462005; RO 462005; Ro-46-2005; Ro 46-2005; Ro46-2005
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO: ~100 mg/mL (~211.2 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.28 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.28 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.28 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1118 mL | 10.5588 mL | 21.1175 mL | |
| 5 mM | 0.4224 mL | 2.1118 mL | 4.2235 mL | |
| 10 mM | 0.2112 mL | 1.0559 mL | 2.1118 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA