| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
Isoxicam is a non-selective inhibitor of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). [2]
X-ray crystallography of the murine COX-2-isoxicam complex reveals that it binds in the cyclooxygenase active site via a novel mode, interacting indirectly with key residues (Arg-120, Tyr-355, Tyr-385, and Ser-530) through two highly coordinated water molecules. It forms a direct hydrogen bond between its 4-hydroxyl group on the thiazine ring and Ser-530 (distance ~2.7 Å). [2] |
|---|---|
| ln Vitro |
The study determined the crystal structure of the murine COX-2-isoxicam complex at 2.0 Å resolution, revealing a novel binding pose. The drug binds in the COX channel in a planar conformation with an intramolecular hydrogen bond between the carboxamide nitrogen and the 4-hydroxyl oxygen of the benzothiazine. It interacts with the hydrophobic channel mainly through van der Waals interactions. A direct hydrogen bond is formed between the 4-hydroxyl group of isoxicam and Ser-530. Two highly ordered water molecules mediate additional polar bridges: one water bridges the carboxamide oxygen and the thiazine nitrogen of isoxicam with Tyr-355 and Arg-120; the other water bridges the heteroatom of the isoxazole ring with Tyr-385 and Ser-530. [2]
The binding of isoxicam induces conformational changes in COX-2, including a rotation of the side chain of Leu-531 (~3.5 Å shift of the Cδ atom) that opens a new hydrophobic pocket, and a movement of helix D (>1.0 Å) to accommodate the benzyl ring of the benzothiazine. [2] |
| Enzyme Assay |
COX Inhibition Assay: Cyclooxygenase inhibition was monitored by O₂ consumption using a Clark-type oxygen-sensitive electrode. The assay solution contained 100 nM purified protein, 2 equivalents of hematin, 100 mM Tris-Cl (pH 8.0), 5 mM phenol, and 2% DMSO. The inhibitor was added and incubated with the protein for 12 min at room temperature, followed by an additional 3 min at 37°C. The reaction was initiated by the addition of arachidonic acid, and oxygen consumption was monitored for 2 min. [2]
Crystallization of mCOX-2-Isoxicam Complex: Purified murine COX-2 (mCOX-2) protein was reconstituted with a 2-fold molar excess of Fe³⁺-protoporphyrin IX. A 10-fold molar excess of isoxicam was added to the protein sample. Crystallization was performed using the hanging drop method. Drops containing 3 µl of the protein-inhibitor complex and 3 µl of crystallization solution (50 mM EPPS, pH 8.0, 120 mM MgCl₂, 22-26% PEG MME-550) were equilibrated against a reservoir of the same solution. Crystals appeared in approximately 5 days and grew to full size in about 3 weeks. Crystals were cryoprotected in a solution containing 50 mM EPPS, pH 8.0, 120 mM MgCl₂, and 28% PEG MME-550 before flash freezing in liquid nitrogen. [2] |
| References |
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| Additional Infomation |
Isoxicam is a monocarboxylic acid amide, with the structural formula of piroxicam, wherein the pyridine-2-yl group is replaced by a 5-methyl-1,2-oxazol-3-yl group. It is a nonsteroidal anti-inflammatory drug (NSAID) that was withdrawn from the market in the 1980s due to its association with Stevens-Johnson syndrome. It can be used as both an NSAID and an antirheumatic drug. It is a benzothiazide compound, belonging to the isoxazole class, and is also a monocarboxylic acid amide. Isoxicam is an NSAID and is not currently marketed in the United States. There are reports of isoxicam's presence in Brassica napus, and relevant data are available.
Isoxicam is a nonselective COX inhibitor belonging to the oxicam class of nonsteroidal anti-inflammatory drugs (NSAIDs). This study provides the first crystal structure determination of an oxicam bound to cyclooxygenase (mCOX-2). The binding mode of isoxicam is novel compared to other NSAIDs, involving a two-water-mediated hydrogen-bonding network and inducing a rotation of Leu-531, which opens a new binding pocket. The structure explains previous structure-activity relationship (SAR) studies of oxicams, which indicated that a methyl group at the 2-position of the benzothiazine ring and a meta-substituted anilide on the carboxamide are optimal for anti-inflammatory activity. [2] |
| Molecular Formula |
C14H13N3O5S
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|---|---|
| Molecular Weight |
335.334
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| Exact Mass |
335.057
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| CAS # |
34552-84-6
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| PubChem CID |
54677972
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| Appearance |
White to off-white solid powder
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
518.8ºC at 760mmHg
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| Melting Point |
265-271℃
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| Flash Point |
267.5ºC
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| Index of Refraction |
1.686
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| LogP |
1.38
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
23
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| Complexity |
627
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
YYUAYBYLJSNDCX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H13N3O5S/c1-8-7-11(16-22-8)15-14(19)12-13(18)9-5-3-4-6-10(9)23(20,21)17(12)2/h3-7,18H,1-2H3,(H,15,16,19)
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| Chemical Name |
4-hydroxy-2-methyl-N-(5-methyl-1,2-oxazol-3-yl)-1,1-dioxo-1λ6,2-benzothiazine-3-carboxamide
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| Synonyms |
BRN 0577221Isoxicam W-8495 W8495W 8495
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| 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)
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| Solubility (In Vitro) |
DMSO : ~5 mg/mL (~14.91 mM)
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|---|---|
| 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 | 2.9821 mL | 14.9107 mL | 29.8214 mL | |
| 5 mM | 0.5964 mL | 2.9821 mL | 5.9643 mL | |
| 10 mM | 0.2982 mL | 1.4911 mL | 2.9821 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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