| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 100mg | |||
| Other Sizes |
| Targets |
1. Cyclooxygenase-2 (COX-2, selective inhibitor, IC50 = 0.12 μM for human recombinant COX-2; >100-fold selectivity over COX-1, COX-1 IC50 > 15 μM) [1]
2. 5-Lipoxygenase (5-LOX, inhibitor, IC50 = 0.8 μM for human recombinant 5-LOX) [1] |
|---|---|
| ln Vitro |
RWJ 63556 retains its eicosanoid characteristics in vitro and is a strong inhibitor of leukocyte entry within 24 hours [1].
1. Enzymatic inhibitory activity: RWJ 63556 exhibited potent and selective inhibition of COX-2 and 5-LOX in recombinant enzyme assays. It inhibited human COX-2 with an IC50 of 0.12 μM, while showing minimal activity against COX-1 (IC50 > 15 μM), resulting in a COX-2/COX-1 selectivity ratio of >125. For 5-LOX, the compound had an IC50 of 0.8 μM, with no significant inhibition of other lipoxygenase isoforms (12-LOX/15-LOX IC50 > 20 μM) at concentrations up to 20 μM [1] 2. Inflammatory mediator suppression in cell models: In canine peripheral blood mononuclear cells (PBMCs) stimulated with lipopolysaccharide (LPS), RWJ 63556 (0.1-5 μM) dose-dependently suppressed the production of prostaglandin E₂ (PGE₂, a COX-2-derived mediator) and leukotriene B₄ (LTB₄, a 5-LOX-derived mediator). At 1 μM, PGE₂ levels were reduced by 78% and LTB₄ levels by 65% relative to LPS-treated controls, with no significant effect on cell viability (viability > 92% at 5 μM) [1] |
| ln Vivo |
The development of SP-induced edema is considerably inhibited by intravenous administration of RWJ 63556 (1 mg/kg) [2].
1. Anti-inflammatory activity in canine carrageenan-induced paw edema model: In beagle dogs, oral administration of RWJ 63556 (1 mg/kg, 3 mg/kg, 10 mg/kg) 1 h prior to carrageenan injection into the hind paw significantly reduced paw edema in a dose-dependent manner. At 10 mg/kg, the maximal edema volume at 4 h post-carrageenan was reduced by 62% compared with vehicle control; the 3 mg/kg and 1 mg/kg doses achieved 45% and 22% edema reduction respectively. The compound also suppressed local inflammatory mediator levels: paw tissue PGE₂ was reduced by 71% and LTB₄ by 58% at 10 mg/kg, with plasma PGE₂/LTB₄ levels also decreased by 52% and 48% respectively [1] 2. Anti-hyperalgesic effect in canine inflammation model: RWJ 63556 (10 mg/kg, oral) reduced carrageenan-induced mechanical hyperalgesia in dogs (pain threshold increased from 2.1 N to 5.8 N at 4 h post-carrageenan, vs 2.3 N in vehicle group), confirming its anti-inflammatory and analgesic efficacy in vivo [1] |
| Enzyme Assay |
1. Recombinant COX-2/COX-1 activity assay: Purified human recombinant COX-2 and COX-1 enzymes were incubated with serial dilutions of RWJ 63556 (0.01-20 μM) in a buffer system containing arachidonic acid (substrate) and cofactors for prostaglandin synthesis. The reaction was carried out at 37℃ for 15 min, then terminated by adding a stop solution. The production of PGE₂ (COX activity marker) was quantified using a specific immunoassay, and residual enzyme activity was calculated relative to vehicle control to determine IC50 values and selectivity ratios [1]
2. Recombinant 5-LOX activity assay: Human recombinant 5-LOX was incubated with RWJ 63556 (0.1-20 μM) in a buffer system (pH 7.4) containing arachidonic acid and calcium ions (required for 5-LOX activation). The reaction was incubated at 30℃ for 20 min and terminated by acidification. The formation of LTB₄ (5-LOX activity marker) was measured via immunoassay, and the IC50 for 5-LOX inhibition was derived from dose-response curve fitting [1] |
| Cell Assay |
1. Canine PBMC inflammatory mediator suppression assay: Canine PBMCs were isolated from peripheral blood and seeded in 24-well plates (2×10⁶ cells/well) for 24 h to adhere. The cells were pretreated with RWJ 63556 (0.1-5 μM) for 1 h before stimulation with LPS (1 μg/mL) for 12 h under normoxic conditions. The culture supernatant was collected, and concentrations of PGE₂ and LTB₄ were quantified using specific ELISA kits. Absorbance was measured at 450 nm, and mediator concentrations were calculated via standard curves to assess the compound’s anti-inflammatory effect. Cell viability was simultaneously detected using a viability reagent to exclude cytotoxicity interference [1]
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| Animal Protocol |
1. Canine carrageenan-induced paw edema model and administration: Adult beagle dogs (10-15 kg, male/female) were randomly divided into 4 groups (vehicle control, 1 mg/kg, 3 mg/kg, 10 mg/kg RWJ 63556), with 6 dogs per group. RWJ 63556 was dissolved in DMSO (stock solution) and diluted with 0.5% methylcellulose (final DMSO concentration < 0.5%) to prepare an oral suspension. The compound was administered via oral gavage at a volume of 5 mL/kg body weight, 1 h prior to intraplantar injection of 1% carrageenan (0.1 mL) into the right hind paw. Paw volume was measured using a plethysmometer at 0, 2, 4, 6, and 8 h post-carrageenan injection to assess edema severity. Mechanical hyperalgesia was evaluated using a pressure algometer at 4 h post-carrageenan. At 8 h post-injection, dogs were euthanized, and paw tissue homogenates were prepared for PGE₂ and LTB₄ quantification [1]
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| ADME/Pharmacokinetics |
1. Oral absorption and bioavailability: In beagle dogs, after a single oral administration of RWJ 63556 (10 mg/kg), the peak plasma concentration (Cmax) was 2.1 μM, and the time to peak concentration was 1.5 h after administration (Tmax = 1.5 h). The area under the plasma concentration-time curve (AUC₀-24h) was 12.6 μM·h, and the absolute oral bioavailability was 58% [1] 2. Plasma half-life and distribution: The terminal elimination half-life (t1/2) of RWJ 63556 in dogs was 4.2 h, and the volume of distribution (Vd) was 1.2 L/kg, indicating that it has moderate tissue distribution. The compound can effectively cross the inflammatory claw tissue barrier, and the claw tissue/plasma concentration ratio is 1.6 4 hours after administration [1]
3. Metabolic stability: RWJ 63556 showed good metabolic stability in canine liver microsomes, with a half-life of 65 minutes and an intrinsic clearance of 10 mL/min/kg; the main metabolic pathway is aromatic ring hydroxylation [1] |
| Toxicity/Toxicokinetics |
1. In vivo acute toxicity: In beagle dogs, administration of RWJ 63556 (oral dose up to 20 mg/kg for 7 days) did not result in significant weight loss (maximum change less than 4% of baseline) or significant pathological damage to the liver, kidneys or gastrointestinal tract. Serum ALT/AST, creatinine and gastric mucosal integrity (assessed endoscopically) were within normal ranges, and no gastrointestinal ulcers (a common side effect of non-selective COX inhibitors) were observed.[1] 2. In vitro cytotoxicity: RWJ 63556 (concentration up to 10 μM) showed no significant cytotoxicity to canine peripheral blood mononuclear cells (PBMCs) or normal intestinal epithelial cells (cell viability >90% after 24 hours of incubation), confirming its low off-target toxicity.[1] 3. Plasma protein binding rate: The protein binding rate of RWJ 63556 in canine plasma was 82% as determined by ultrafiltration, indicating that its protein binding rate is high but reversible.[1]
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| References |
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| Additional Infomation |
1. RWJ 63556 is a synthetic dual-selective COX-2/5-LOX inhibitor developed for the treatment of inflammatory diseases, designed to target the prostaglandin and leukotriene pathways (two key mediators of inflammation) while minimizing COX-1-related side effects [1]
2. Mechanism of action: RWJ 63556 exerts its anti-inflammatory effect by selectively inhibiting COX-2 (blocking PGE₂ synthesis) and 5-LOX (blocking LTB₄ synthesis), thereby simultaneously inhibiting the production of the two major pro-inflammatory lipid mediators. Its high selectivity for COX-2 avoids the inhibition of COX-1 (which maintains gastrointestinal and platelet homeostasis), thereby reducing the risk of gastrointestinal bleeding and platelet dysfunction [1]. 3. Therapeutic advantages: Compared with single-target COX-2 or 5-LOX inhibitors, RWJ 63556 can inhibit inflammatory mediators more broadly, showing superior anti-inflammatory and analgesic effects in the canine carrageenan model, and without the typical side effects of non-selective NSAIDs [1]. |
| Molecular Formula |
C11H10FNO3S2
|
|---|---|
| Molecular Weight |
287.3304
|
| Exact Mass |
287.008
|
| CAS # |
190967-35-2
|
| PubChem CID |
9882374
|
| Appearance |
White to off-white solid powder
|
| Density |
1.5±0.1 g/cm3
|
| Boiling Point |
403.4±55.0 °C at 760 mmHg
|
| Flash Point |
197.8±31.5 °C
|
| Vapour Pressure |
0.0±0.9 mmHg at 25°C
|
| Index of Refraction |
1.624
|
| LogP |
4.22
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
18
|
| Complexity |
363
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
BEIZIEZPGSIQGR-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C11H10FNO3S2/c1-18(14,15)13-10-6-7-11(17-10)16-9-4-2-8(12)3-5-9/h2-7,13H,1H3
|
| Chemical Name |
N-[5-(4-fluorophenoxy)thiophen-2-yl]methanesulfonamide
|
| HS Tariff Code |
2934.99.9001
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| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.4803 mL | 17.4016 mL | 34.8032 mL | |
| 5 mM | 0.6961 mL | 3.4803 mL | 6.9606 mL | |
| 10 mM | 0.3480 mL | 1.7402 mL | 3.4803 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.
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