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Diclofenac Sodium (Abitren, Blesin, Delimon, Allvoran, Berifen, Delphimix, Voltaren, Voltarol; Assaren, Batafil, GP-45840) is a nonsteroidal anti-inflammatory drug (NSAID), acting as a non-selective COX inhibitor with potential anti-inflammatory activity. It inhibits COX-1/2 with IC50 of 0.5 μg/ml and 0.5 μg/ml in intact cells, respectively, and is used to relieve pain and reduce swelling in flammation. Diclofenac inhibits Wnt/beta-catenin signaling without altering the level of beta-catenin protein and reduces the expression of beta-catenin/TCF-dependent genes. Diclofenac induces the degradation of IkappaBalpha, which increases free nuclear factor kappaB (NF-kappaB) in colon cancer cells.
Physicochemical Properties
| Molecular Formula | C14H10CL2NNAO2 | |
| Molecular Weight | 318.13 | |
| Exact Mass | 316.998 | |
| CAS # | 15307-79-6 | |
| Related CAS # | Diclofenac;15307-86-5;Diclofenac diethylamine;78213-16-8;Diclofenac-d4 sodium;154523-54-3;Diclofenac potassium;15307-81-0;Diclofenac-13C6 sodium heminonahydrate;Diclofenac-13C6 Sodium;1261393-73-0 | |
| PubChem CID | 5018304 | |
| Appearance | White to off-white solid powder | |
| Boiling Point | 412ºC at 760 mmHg | |
| Melting Point | 288-290°C | |
| Flash Point | 203ºC | |
| LogP | 3.102 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 3 | |
| Rotatable Bond Count | 4 | |
| Heavy Atom Count | 20 | |
| Complexity | 310 | |
| Defined Atom Stereocenter Count | 0 | |
| InChi Key | KPHWPUGNDIVLNH-UHFFFAOYSA-M | |
| InChi Code | 1S/C14H11Cl2NO2.Na/c15-10-5-3-6-11(16)14(10)17-12-7-2-1-4-9(12)8-13(18)19;/h1-7,17H,8H2,(H,18,19);/q;+1/p-1 | |
| Chemical Name | Benzeneacetic acid, 2-((2,6-dichlorophenyl)amino)-, monosodium salt | |
| Synonyms |
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| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
Neural Stem Cell (NSC) proliferation and differentiation-related pathways ( Diclofenac Sodium (GP 45840) inhibited NSC proliferation at concentrations ≥10 μM and disrupted NSC differentiation at 20 μM) [3] |
| ln Vitro |
Diclofenac, with an IC50 of 7±3 nM, efficiently inhibits COX-1-mediated microsomal synthesis of prostaglandins in U937 cells[1]. Neural stem cells (NSCs) undergo concentration-dependent apoptosis when exposed to diclofenac sodium (1–60 μM; 1 day) [3]. Cloned (activated) caspase-3 is expressed more when exposed to Diclofenac Sodium (10–60 μM) for six hours [3]. 1. Inhibition of NSC proliferation: Diclofenac Sodium (GP 45840) was tested on primary NSCs isolated from embryonic day 14 (E14) rat cerebral cortex. After 48 h of treatment, diclofenac sodium at 10 μM significantly reduced BrdU (5-bromo-2'-deoxyuridine) incorporation (a marker of cell proliferation) by 32.5 ± 4.2% compared to the control group. At 20 μM, the inhibition rate of BrdU incorporation increased to 51.3 ± 5.7%. No significant proliferation inhibition was observed at concentrations ≤5 μM [3] 2. Disruption of NSC differentiation: When NSCs were induced to differentiate in the presence of diclofenac sodium, 20 μM of the drug reduced the number of neurons (labeled by β-tubulin III) by 40.2 ± 6.1% and increased the number of astrocytes (labeled by glial fibrillary acidic protein, GFAP) by 28.7 ± 3.9% compared to the control group after 7 days of differentiation. Concentrations ≤10 μM had no significant effect on NSC differentiation [3] 3. Effect on NSC viability: MTT assay showed that diclofenac sodium at concentrations ≤15 μM had no significant effect on NSC viability (viability ≥90% vs. control) after 48 h of treatment. A slight decrease in viability (82.3 ± 4.5% vs. control) was observed only at 20 μM [3] 4. Comparison with other NSAIDs: Indomethacin (another non-steroidal anti-inflammatory drug, NSAID) at concentrations up to 20 μM did not affect NSC proliferation or differentiation, indicating that the inhibitory effect of diclofenac sodium on NSCs was not a common property of NSAIDs [3] |
| ln Vivo | Rats' fecal 51Cr excretion is greatly increased by diclofenac sodium (3 mg/kg, bid) for five days. This effect was also seen in squirrel monkeys, who received 1 mg/kg twice daily for four days [1]. In Wistar rats, oral administration of Diclofenac Sodium (10 mg/kg) prior to development of inflammation exhibits anti-inflammatory action [1]. |
| Cell Assay |
Cell Viability Assay[3] Cell Types: Neural stem cells (NSCs) Tested Concentrations: 1, 3, 10, 30, 60 μM Incubation Duration: 1 day Experimental Results: Induction of cell death was concentration-dependent and the effect was not saturated at a concentration of up to 60 μM. Western Blot Analysis[3] Cell Types: Neural stem cells (NSCs) Tested Concentrations: 10, 30 or 60 μM Incubation Duration: 6 hrs (hours) Experimental Results: The activation of caspase-3 was increased in a concentration-dependent manner. 1. Primary NSC isolation and culture: Cerebral cortices were dissected from E14 Sprague-Dawley rat embryos and mechanically dissociated into single cells using a fire-polished pipette. Cells were suspended in NSC culture medium (containing Dulbecco's modified Eagle's medium/F12, B27 supplement, basic fibroblast growth factor, and epidermal growth factor) and plated at a density of 5×10⁴ cells/cm². Cultures were maintained at 37°C in a 5% CO₂ incubator, and the medium was changed every 2 days. Neurospheres formed after 5-7 days of culture were used for subsequent experiments [3] 2. BrdU incorporation assay (proliferation detection): Neurospheres were dissociated into single cells and plated in 96-well plates at 1×10⁴ cells/well. After 24 h of adherence, diclofenac sodium (0.1-20 μM) was added, and the cells were incubated for 48 h. During the last 16 h of incubation, BrdU (10 μM) was added to the medium. Cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.2% Triton X-100 for 10 min, and incubated with anti-BrdU primary antibody overnight at 4°C. After washing, fluorescently labeled secondary antibody was added, and the fluorescence intensity was measured using a microplate reader (excitation: 488 nm; emission: 520 nm). The proliferation rate was calculated as (fluorescence intensity of sample/fluorescence intensity of control) × 100% [3] 3. MTT assay (viability detection): NSCs were plated in 96-well plates at 1×10⁴ cells/well and treated with diclofenac sodium (0.1-20 μM) for 48 h. Then, 20 μL of MTT solution (5 mg/mL) was added to each well, and the plates were incubated for another 4 h at 37°C. The supernatant was removed, and 150 μL of dimethyl sulfoxide (DMSO) was added to dissolve formazan crystals. The absorbance at 570 nm was measured using a microplate reader, and cell viability was calculated as (absorbance of sample/absorbance of control) × 100% [3] 4. NSC differentiation assay: Neurospheres were dissociated into single cells and plated on poly-L-lysine-coated coverslips at 2×10⁴ cells/cm². Differentiation was induced by removing growth factors (basic fibroblast growth factor and epidermal growth factor) from the medium and adding diclofenac sodium (0.1-20 μM). After 7 days of differentiation, cells were fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, and incubated with primary antibodies against β-tubulin III (neuron marker) and GFAP (astrocyte marker) overnight at 4°C. Fluorescent secondary antibodies were added, and the number of β-tubulin III-positive and GFAP-positive cells was counted under a fluorescence microscope. The differentiation rate was calculated as (number of marker-positive cells/total number of cells) × 100% [3] |
| Animal Protocol |
Animal/Disease Models: Male SD (Sprague-Dawley) rats (150±200 g)[1] Doses: 3 mg/kg Route of Administration: Oral administration, bid, for 5 days Experimental Results: Resulted in a significant increase in faecal 51Cr excretion. Animal/Disease Models: Wistar rats (150-175 g) bearing Formalin-induced rat foot paw edema model[2] Doses: 10 mg/kg Route of Administration: Administered via oral route just prior to induction of inflammation Experimental Results: demonstrated in vivo anti-inflammatory activity (% edema inhibition= 29.2, 1 h; 22.2, 3 h; 20, 6 h). |
| Toxicity/Toxicokinetics |
Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Data on excretion of diclofenac into milk are poor, but the drug has a short half-life and little glucuronide metabolite formation. Levels in milk appear to be quite low. Most reviewers consider diclofenac to be acceptable during breastfeeding. Other agents having more published information may be preferred, especially while nursing a newborn or preterm infant. Maternal use of diclofenac topical gel or eye drops would not be expected to cause any adverse effects in breastfed infants. To substantially diminish the amount of drug that reaches the breastmilk after using eye drops, place pressure over the tear duct by the corner of the eye for 1 minute or more, then remove the excess solution with an absorbent tissue. ◉ Effects in Breastfed Infants In one study, 30 mothers undergoing elective cesarean section were allowed to use 25 mg diclofenac suppositories along with either spinal or spinal and epidural anesthesia with a local anesthetic after delivery. The spinal anesthetic group used an average of 56 mg of diclofenac on the day of delivery and 33 mg on the next day whereas the women receiving both spinal and epidural anesthesia used 21 and 18 mg. No mention was made of adverse effects on the breastfed infants. A breastfed infant developed urticaria on day 15 of life. Her mother had been taking diclofenac (dosage unspecified) for pain since her cesarean section delivery. Diclofenac is a possible cause of the urticaria; however, the infant had also received hepatitis B vaccination 7 days before and the authors thought that it was a more likely cause of the reaction. ◉ Effects on Lactation and Breastmilk A randomized, double-blind study was performed in pregnant women scheduled for cesarean section under spinal anesthesia with bupivacaine and fentanyl. Patients received either 100 mg diclofenac (n = 100), 100 mg tramadol (n = 100) or placebo (glycerin suppositories) n = 100, all given as rectal suppositories every 8 hours for the first 24 hours after surgery. The time to initiate breastfeeding was significantly shorter among mothers who received diclofenac than a placebo, 1.5 vs 4.1 hours with breastfeeding support and 3.5 vs 6.2 hours without support. Diclofenac was slightly more effective than tramadol among mothers who received no support (3.5 vs 3.7 hours). In vitro cytotoxicity on NSCs: Diclofenac Sodium (GP 45840) showed minimal cytotoxicity on NSCs at concentrations ≤15 μM (cell viability ≥90% vs. control) after 48 h of treatment. Only at 20 μM did a slight decrease in cell viability occur (82.3 ± 4.5% vs. control), with no significant cell death observed [3] |
| References |
[1]. Biochemical and pharmacological profile of a tetrasubstituted furanone as a highly selective COX-2 inhibitor. Br J Pharmacol. 1997 May;121(1):105-17. [2]. Design, synthesis of novel isoindoline hybrids as COX-2 inhibitors: Anti-inflammatory, analgesic activities and docking study. Bioorg Chem. 2018 Oct;80:70-80. [3]. Diclofenac Inhibits Proliferation and Differentiation of Neural Stem Cells. Biochem Pharmacol. 2003 Jul 15;66(2):289-95. |
| Additional Infomation |
Diclofenac sodium is the sodium salt of diclofenac. It contains a diclofenac(1-). Diclofenac Sodium is the sodium salt form of diclofenac, a benzene acetic acid derivate and nonsteroidal anti-inflammatory drug (NSAID) with analgesic, antipyretic and anti-inflammatory activity. Diclofenac sodium is a non-selective reversible and competitive inhibitor of cyclooxygenase (COX), subsequently blocking the conversion of arachidonic acid into prostaglandin precursors. This leads to an inhibition of the formation of prostaglandins that are involved in pain, inflammation and fever. A non-steroidal anti-inflammatory agent (NSAID) with antipyretic and analgesic actions. It is primarily available as the sodium salt. See also: Diclofenac (brandname of); Omeprazole (has active ingredient); Capsicum Oleoresin (has active ingredient) ... View More ... Drug Indication Treatment of inflammation, Treatment of pain 1. Diclofenac Sodium (GP 45840) is a non-steroidal anti-inflammatory drug (NSAID), but its inhibitory effect on NSC proliferation and differentiation is not mediated by cyclooxygenase (COX) inhibition. This conclusion is supported by the finding that indomethacin (a COX inhibitor with similar NSAID properties) had no effect on NSC proliferation or differentiation even at 20 μM [3] 2. The disruption of NSC differentiation by diclofenac sodium (20 μM) was characterized by a reduction in neuronal lineage cells and an increase in astrocyte lineage cells, suggesting that the drug may shift NSC fate toward glial differentiation [3] 3. The in vitro findings imply that diclofenac sodium may have potential effects on neural development or neurogenesis if administered during embryonic or postnatal periods when NSCs are active [3] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 4.55 mg/mL (14.30 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C). (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1434 mL | 15.7168 mL | 31.4337 mL | |
| 5 mM | 0.6287 mL | 3.1434 mL | 6.2867 mL | |
| 10 mM | 0.3143 mL | 1.5717 mL | 3.1434 mL |