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Research Area
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Description
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Inflammation |
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Biological Activity
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Description
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Ibuprofen (Advil, Dolgesic) is an anti-inflammatory inhibitor targeting COX-1 and COX-2 with IC50 of 13 μM and 370 μM, respectively. |
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Targets
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COX-1 |
COX-2 |
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IC50 |
13 μM |
370 μM [1] |
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In Vitro
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Ibuprofen works by inhibiting the enzyme cyclooxygenase COX-1 and COX-2, which convert arachidonic acid to prostaglandin H2 (PGH2). Its action is similar to aspirin, indomethacin and all other NSAIDs in intact cells, broken cells, and purified enzyme preparations. [1] Ibuprofen inhibits the constitutive activation of NF-κB and IKKα in the androgen-independent prostate tumor cells PC-3 and DU-145. It sensitizes prostate cells to ionizing radiation and blocks stimulated activation of NF-κB following exposure to TNFα or ionizing radiation in the androgen-sensitive prostate tumor cell line LNCaP. Both of these cannot be attributed directly to inhibition of IκB-α kinase but to inhibition of an upstream regulator of IKKα. [2] Ibuprofen exerts an anticancer effect by reducing survival of cancer cells. Ibuprofen is more efficacious than aspirin and acetaminophen, and comparable with (R)-flurbiprofen and indomethacin in induction of p75NTR protein (a tumor and metastasis suppressor) expression in cell lines from bladder and other organs. [3] |
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In Vivo
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Ibuprofen reacts with the heme group of cyclooxygenase to prevent arachidonic acid conversion. Prior exposure to Ibuprofen in vivo protects cyclooxygenase completely from the irreversible effects of aspirin in platelets. [4] Ibuprofen treatment is effective in attenuating joint inflammation and early articular cartilage degeneration in the adult female Sprague-Dawley rat model induced by high-repetition and high-force (HRHF) task. It dose this by blocking the increases in serum C1 and 2C (a biomarker of collagen I and II degradation) as well as the ratio of collagen degradation to synthesis (C1, 2C/CPII, the latter a biomarker of collage type II synthesis) induced by HRHF. [5] |
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Clinical Trials
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Phase IV completed in evaluating the analgesic effect of Ibuprofen 400, 600 and 800 mg, Paracetamol 500 and 1000 mg, and Paracetamol 1000 mg plus 60 mg Codeine on acute pain after third molar surgery. |
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Features
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Ibuprofen is a core medicine in the World Health Organization's "WHO Model List of Essential Medicines", which is a list of the minimum medical requirements for a basic healthcare system. |
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Protocol
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Kinase Assay
[1]
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| Radiochemical enzyme assays for COX-1 and COX-2 |
10 μL of purified COX-1 (0.7-0.8 μg) or COX-2 (3.0 units, 0.3μg) is activated with 50 μL of cofactor solution [l-epinephrine (1.3 mg/mL), reduced glutathione (0.3 mg/mL), and hematin (1.3 mg/mL) in oxygen-free Tris-HCl buffer (pH 8.0)]. The enzyme solution (60 μL) is added to Ibuprofen solutions or DMSO (20 μL) after [14C]arachidonic acid is added in 0.2 mL eight-strip test tubes and preincubated 10 minutes on ice. Samples are incubated for 15 minutes at 37 °C, after which the reaction is terminated by addition of 10 μL of 2 M HCl and 5 μL of carrier solution (PGE2 and PGF2α, 0.2 μg/mL of each in EtOH). The unmetabolized arachidonic acid is separated from the prostaglandin products by column chromatography and eluted with n-hexane-dioxane-glacial acetic acid (70:30:1). The prostaglandin products are then eluted with EtOAc-MeOH (85:15), and the samples are counted in a Packard scintillation spectrometer. IC50 values are obtained by linear regression analysis. |
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Cell Assay
[3]
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| Cell Lines |
Bladder epithelial cell line T24, RT-4 transitional cell papilloma bladder cell line, 5637 primary carcinoma bladder cell line, HCT-116, MDAMB231, MCF7, HEK293, A549, SKOV3 and DU145 |
| Concentrations |
Dissolved in DMSO at a concentration of 200 mM for making the stock solution, final concentration ~2 mM |
| Incubation Time |
48 hours |
| Methods |
Each cell line is incubated with Ibuprofen of various concentrations for 48 hours. Cell survival is estimated by the MTT assay, and cell death is determined by Hoechst staining used to distinguish between intact cell nuclei and fragmented nuclei undergoing cell death. Cells are lysed and analyzed by western blotting for detection of the p75NTR protein. |
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Animal Study
[5]
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| Animal Models |
Female Sprague-Dawley rats with joint inflammation induced by high-repetition and high-force (HRHF) tasks |
| Formulation |
Dissolved in DMSO and diluted in saline |
| Doses |
45 mg/kg |
| Administration |
Taken orally every day |
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References |
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[1] Noreen Y, et al. J Nat Prod, 1998, 61(1), 2-7.
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[2] Palayoor ST, et al. Oncogene, 1999, 18(51), 7389-7394.
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[3] Khwaja F, et al. Cancer Res, 2004, 64(17), 6207-6213.
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[4] Rao GH, et al. Arteriosclerosis, 1983, 3(4), 383-388.
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[5] Driban JB, et al. J Biomed Biotechnol, 2011, 2011, 691412-691428.
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