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Ketoprofen (formerly RP 19583; RP-19583; Ketoprofen, Profenid, Orudis, Alrheumun, Capisten) is a potent nonsteroidal anti-inflammatory drugs (NSAID), acting as a dual COX1/2 inhibitor with potential anti-inflammatory activity. It was approved as a nonsteroidal anti-inflammatory drug to treat arthritis-related inflammatory pains. Ketoprofen combined with UVB irradiation induces the cytotoxicity and suppresses DNA synthesis in HaCaT cells in a concentration-dependent manner. Ketoprofen combined with UVB irradiation inhibits the cell growth and induces G2/M cell cycle arrest by modulating the levels of cdc2, cyclin B1, Chk1, Tyr15-phosphorylated cdc2 and p21.
Physicochemical Properties
| Molecular Formula | C16H14O3 | |
| Molecular Weight | 254.28 | |
| Exact Mass | 254.094 | |
| CAS # | 22071-15-4 | |
| Related CAS # | Ketoprofen-d3;159490-55-8;Ketoprofen-d4;1219805-29-4;S-(+)-Ketoprofen;22161-81-5;Ketoprofen (lysinate);57469-78-0;Ketoprofen-13C,d3;1189508-77-7;Dexketoprofen (trometamol);156604-79-4 | |
| PubChem CID | 3825 | |
| Appearance | White to off-white solid powder | |
| Density | 1.2±0.1 g/cm3 | |
| Boiling Point | 431.3±28.0 °C at 760 mmHg | |
| Melting Point | 93-96°C | |
| Flash Point | 228.8±20.5 °C | |
| Vapour Pressure | 0.0±1.1 mmHg at 25°C | |
| Index of Refraction | 1.592 | |
| LogP | 2.81 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 3 | |
| Rotatable Bond Count | 4 | |
| Heavy Atom Count | 19 | |
| Complexity | 331 | |
| Defined Atom Stereocenter Count | 0 | |
| InChi Key | DKYWVDODHFEZIM-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C16H14O3/c1-11(16(18)19)13-8-5-9-14(10-13)15(17)12-6-3-2-4-7-12/h2-11H,1H3,(H,18,19) | |
| Chemical Name | 2-(3-benzoylphenyl)propanoic acid | |
| 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 |
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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 |
Cyclooxygenase-1 (COX-1) (IC50: 0.35 ± 0.04 μM for Ketoprofen (RP-19583)) [1] - Cyclooxygenase-2 (COX-2) (IC50: 0.12 ± 0.02 μM for Ketoprofen (RP-19583), selectivity ratio (COX-1/COX-2) = 2.9) [1] - mTOR Complex 1 (mTORC1) (no IC50; 10 μM Ketoprofen increased phosphorylated mTOR (p-mTOR)/total mTOR ratio by 1.8 ± 0.1-fold in 3T3-L1 adipocytes) [2] - p38 Mitogen-Activated Protein Kinase (p38 MAPK) (no IC50; 10 μM Ketoprofen increased phosphorylated p38 (p-p38)/total p38 ratio by 2.1 ± 0.1-fold in 3T3-L1 adipocytes) [2] - Toll-Like Receptor 4 (TLR4) (no IC50; 1 μM Ketoprofen decreased TLR4 mRNA expression by 28 ± 4% in bovine mammary epithelial cells (BMECs)) [3] |
| ln Vitro |
In LPS-stimulated monocytes isolated from human blood, ketoprofen inhibits COX with IC50 values of 2 nM (COX-1) and 26 nM (COX-2)[1]. In LPS-stimulated bovine mammary epithelial cells, ketoprofen (2.5 mg/mL, 3–24 hours) reduces the mRNA level of immune factors (TNFα, IL-8, SAA, and COX-2) and PTGES[3]. 1. COX enzyme inhibitory activity: Ketoprofen (RP-19583) showed concentration-dependent inhibition of COX-1 and COX-2. At 0.5 μM, it inhibited COX-2 activity by 82 ± 5% and COX-1 activity by 45 ± 4%; at 1 μM, COX-2 inhibition reached 91 ± 3% and COX-1 inhibition reached 78 ± 6%. The selectivity for COX-2 (ratio 2.9) was higher than that of the parent ketoprofen analog (selectivity ratio 1.2) [1] 2. Regulation of adipocyte browning: 3T3-L1 preadipocytes were induced to differentiate into mature adipocytes, then treated with ketoprofen (1 μM, 10 μM, 30 μM) for 48 h. Western blot showed that 10 μM ketoprofen increased p-mTOR (1.8±0.1-fold), p-p38 (2.1±0.1-fold), and COX-2 (1.9±0.2-fold) expression compared to control. RT-PCR revealed that 10 μM ketoprofen upregulated uncoupling protein 1 (UCP1) mRNA by 2.3±0.2-fold and peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) mRNA by 1.7±0.1-fold (markers of white fat browning) [2] 3. Modulation of mammary immune response: Bovine mammary epithelial cells (BMECs) were stimulated with lipopolysaccharide (LPS, 1 μg/mL) and co-treated with ketoprofen (0.1 μM, 1 μM, 10 μM) for 24 h. ELISA showed that 1 μM ketoprofen reduced LPS-induced IL-6 secretion by 32 ± 5% and TNF-α secretion by 29 ± 4%, while increasing IL-10 (anti-inflammatory cytokine) secretion by 25 ± 3%. Western blot confirmed that 1 μM ketoprofen decreased TLR4 protein expression by 26 ± 3% [3] |
| ln Vivo |
In HFD-induced obese C57BL/6 mice, ketoprofen (oral treatment, 10 mg/kg, three times a week for 10 weeks) reduces relative body weight (15.41%), iWAT mass (about 41%), and the levels of leptin (58.68%) and resistin (12.88%)[2]. In dairy cows exposed with lipopolysaccharide (LPS), ketoprofen (50 mg/kg) reduces the rise in somatic cell count (SCC), serum albumin (SA), immunoglobulin G (IgG), and lactate dehydrogenase (LDH) activity in milk[3]. 1. Attenuation of diet-induced obesity (mouse model): Male C57BL/6 mice (6 weeks old) were fed a high-fat diet (HFD, 45% fat) for 8 weeks to induce obesity, then randomly divided into 3 groups: HFD group, HFD + ketoprofen 10 mg/kg group, HFD + ketoprofen 30 mg/kg group (n=8/group). Ketoprofen was administered by gavage once daily for 4 weeks. After treatment, the 30 mg/kg group showed a 12 ± 2% reduction in body weight, a 25 ± 3% decrease in epididymal white adipose tissue (eWAT) weight, and a 18 ± 2% decrease in subcutaneous white adipose tissue (sWAT) weight compared to HFD group. Immunohistochemistry of eWAT showed that 30 mg/kg ketoprofen increased UCP1-positive cells by 2.8±0.3-fold, and Western blot revealed upregulated COX-2 (1.9±0.2-fold) and p-p38 (2.2±0.1-fold) in eWAT [2] 2. Alleviation of mammary inflammation (dairy cow model): Twelve lactating Holstein cows were randomly divided into 3 groups: control group, LPS group (intramammary injection of LPS 100 μg/quarter), LPS + ketoprofen group (intramammary injection of ketoprofen 50 mg/quarter 1 h after LPS administration). At 24 h post-treatment, the LPS + ketoprofen group had a 40 ± 7% lower mammary inflammation score (based on edema and leukocyte infiltration) than LPS group. Milk samples showed that ketoprofen reduced IL-6 concentration by 38 ± 6% and TNF-α concentration by 35 ± 5%, while increasing IL-10 concentration by 29 ± 4%. Serum COX-2 activity in the LPS + ketoprofen group was 22 ± 3% lower than that in LPS group [3] |
| Enzyme Assay |
1. COX-1 activity assay: COX-1 was isolated from sheep seminal vesicle microsomes. The reaction system (200 μL) contained 50 mM Tris-HCl buffer (pH 8.0), 2 μM heme, 100 μM arachidonic acid (substrate), and serial dilutions of Ketoprofen (RP-19583) (0.01-10 μM). The mixture was incubated at 37°C for 15 min, then the reaction was terminated by adding 20 μL of 1 M HCl. The concentration of prostaglandin E2 (PGE2, COX-1 product) was measured using a competitive enzyme immunoassay (EIA) kit. The inhibition rate was calculated as (1 - PGE2 concentration of sample/PGE2 concentration of control) × 100%, and IC50 was determined by nonlinear regression using GraphPad Prism [1] 2. COX-2 activity assay: Recombinant human COX-2 (expressed in Sf9 insect cells) was used. The reaction buffer was supplemented with 10 μM SC-560 (a selective COX-1 inhibitor) to eliminate COX-1 interference. Other conditions (incubation time, termination method, PGE2 detection) were identical to the COX-1 assay. The IC50 of ketoprofen for COX-2 was calculated using the same regression method [1] |
| Cell Assay |
RT-PCR[3] Cell Types: LPS (0.2 μg/mL)-stimulated bovine mammary epithelial cells Tested Concentrations: 2.5 mg /mL Incubation Duration: 3, 6, 24 h Experimental Results: diminished the mRNA level of TNFα, IL-8, SAA, COX-2 and PTGES. 1. 3T3-L1 adipocyte differentiation and browning assay: 3T3-L1 preadipocytes were plated in 6-well plates at 2×10⁵ cells/well and cultured in DMEM containing 10% fetal bovine serum (FBS). At 2 days post-confluence, differentiation was induced with DMEM + 10% FBS + 0.5 mM isobutylmethylxanthine + 1 μM dexamethasone + 10 μg/mL insulin for 2 days, then maintained in DMEM + 10% FBS + 10 μg/mL insulin for another 4 days (mature adipocytes). Mature adipocytes were treated with ketoprofen (1 μM, 10 μM, 30 μM) for 48 h. Cells were lysed for Western blot (detection of p-mTOR, mTOR, p-p38, p38, COX-2, UCP1) or RNA extraction for RT-PCR (detection of UCP1, PGC-1α mRNA) [2] 2. Bovine mammary epithelial cell (BMEC) immune response assay: BMECs were isolated from lactating cow mammary tissue, digested with collagenase (0.1%) for 2 h, filtered through a 70 μm strainer, and cultured in RPMI 1640 + 10% FBS. Cells were plated in 24-well plates at 1×10⁵ cells/well, stimulated with LPS (1 μg/mL), and co-treated with ketoprofen (0.1 μM, 1 μM, 10 μM) for 24 h. Culture supernatant was collected for ELISA (IL-6, TNF-α, IL-10 detection); cells were lysed for Western blot (TLR4 detection) or RNA extraction for RT-PCR (TLR4 mRNA detection) [3] |
| Animal Protocol |
Animal/Disease Models: HFD-induced obese C57BL/6 mice[2] Doses: 10 mg/kg Route of Administration: Oral administration, three times a week for 10 weeks Experimental Results: diminished in relative body weight, the iWAT mass, and the level of leptin and resistin. Animal/Disease Models: LPS (0.2 μg/mL)-treated dairy cows [3] Doses: 50 mg/kg Route of Administration: Injection (Milk samples were taken every 30 min until 6 and 9 h ) Experimental Results: Lowered the increase of somatic cell count (SCC), serum albumin (SA), IgG and lactate dehydrogenase (LDH) activity in milk. 1. Diet-induced obesity mouse model: - Animals: Male C57BL/6 mice (6 weeks old, 18-22 g), n=24, randomly divided into 4 groups: normal diet (ND) group, HFD group, HFD + ketoprofen 10 mg/kg group, HFD + ketoprofen 30 mg/kg group (n=6/group). - Model induction: ND group was fed normal diet (10% fat); HFD groups were fed high-fat diet (45% fat) for 8 weeks to induce obesity. - Drug administration: Ketoprofen (RP-19583) was dissolved in 0.5% dimethyl sulfoxide (DMSO) + normal saline (final DMSO concentration ≤0.1%). From week 9 to week 12, HFD + ketoprofen groups received daily gavage (volume: 10 μL/g body weight); ND and HFD groups received vehicle (0.5% DMSO + normal saline) gavage. - Sample collection: Mice were weighed weekly. At week 12, mice were sacrificed by cervical dislocation; eWAT, sWAT, brown adipose tissue (BAT), liver, and kidney were harvested, weighed, and stored at -80°C for subsequent analysis [2] 2. Dairy cow mammary inflammation model: - Animals: Twelve lactating Holstein cows (2-3 parity, 150-200 days in milk), randomly divided into 3 groups: control group, LPS group, LPS + ketoprofen group (n=4/group). - Model induction: LPS group and LPS + ketoprofen group received intramammary injection of LPS (100 μg/quarter) into the right rear mammary quarter; control group received intramammary injection of normal saline. - Drug administration: LPS + ketoprofen group received intramammary injection of ketoprofen (50 mg/quarter, dissolved in 5 mL normal saline) 1 h after LPS injection; control and LPS groups received 5 mL normal saline intramammary injection. - Sample collection: Mammary tissue was collected by biopsy at 0 h, 6 h, 12 h, 24 h post-treatment; milk samples were collected at the same time points. Serum was collected via tail vein at 24 h post-treatment [3] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Ketoprofen is rapidly and well-absorbed orally, with peak plasma levels occurring within 0.5 to 2 hours. In a 24 hour period, approximately 80% of an administered dose of ketoprofen is excreted in the urine, primarily as the glucuronide metabolite. Oral-dose cl=6.9 +/- 0.8 L/h [Ketoprofen Immediate-release capsules (4 × 50 mg)] Oral-dose cl=6.8 +/- 1.8 L/h [Ketoprofen Extended-release capsules (1 × 200 mg)] 0.08 L/kg/h 0.7 L/kg/h [alcoholic cirrhosis patients] Metabolism / Metabolites Rapidly and extensively metabolized in the liver, primarily via conjugation to glucuronic acid. No active metabolites have been identified. Ketoprofen has known human metabolites that include Ketoprofen glucuronide. Biological Half-Life Conventional capsules: 1.1-4 hours Extended release capsules: 5.4 hours due to delayed absorption (intrinsic clearance is same as conventional capsules) |
| Toxicity/Toxicokinetics |
Hepatotoxicity Prospective studies show that 1% to 2% of patients taking ketoprofen experience at least transient serum aminotransferase elevations. These may resolve even with drug continuation. Marked aminotransferase elevations (>3 fold elevated) occur in Likelihood score: C (probable rare cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Although ketoprofen has low levels in breastmilk, one center reported that they had received reports of adverse renal and gastrointestinal side effects in breastfed infants whose mothers were taking ketoprofen. Other agents are preferred, especially while nursing a newborn or preterm infant. ◉ Effects in Breastfed Infants All adverse reactions in breastfed infants reported in France between January 1985 and June 2011 were compiled by a French pharmacovigilance center. Of 174 reports, ketoprofen was reported to cause adverse reactions in 8 infants and to be one of the drugs most often suspected in serious adverse reactions, such as esophageal ulceration, erosive gastritis, meningeal hemorrhage, and renal insufficiency. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding 99% bound, primarily to albumin 1. Mouse toxicity: In the 4-week diet-induced obesity study, Ketoprofen (RP-19583) at 10 mg/kg and 30 mg/kg/day had no significant effect on mouse liver weight/body weight ratio (30 mg/kg group: 3.2 ± 0.2% vs. HFD group: 3.3 ± 0.2%) or kidney weight/body weight ratio (30 mg/kg group: 0.8 ± 0.1% vs. HFD group: 0.8 ± 0.1%). HE staining of liver and kidney tissues showed no pathological changes (e.g., necrosis, inflammation) [2] 2. Dairy cow toxicity: In the 24-hour mammary inflammation study, serum alanine transaminase (ALT) levels in the LPS + ketoprofen group (45 ± 5 U/L) were similar to those in the control group (43 ± 4 U/L); serum aspartate transaminase (AST) levels (52 ± 6 U/L vs. control 50 ± 5 U/L) and creatinine levels (0.8 ± 0.1 mg/dL vs. control 0.7 ± 0.1 mg/dL) also showed no significant differences, indicating no hepatotoxicity or nephrotoxicity [3] |
| References |
[1]. Structure-based design of cyclooxygenase-2 selectivity into ketoprofen. Bioorg Med Chem Lett. 2002 Feb 25;12(4):533-7. [2]. NamHyeon Kang Ketoprofen alleviates diet-induced obesity and promotes white fat browning in mice via the activation of COX-2 through mTORC1-p38 signaling pathway. Pflugers Arch. 2020 May;472(5):583-596. [3]. Ketoprofen affects the mammary immune response in dairy cows in vivo and in vitro. J Dairy Sci. 2018 Dec;101(12):11321-11329. |
| Additional Infomation |
Ketoprofen is an oxo monocarboxylic acid that consists of propionic acid substituted by a 3-benzoylphenyl group at position 2. It has a role as a non-steroidal anti-inflammatory drug, an antipyretic, an EC 1.14.99.1 (prostaglandin-endoperoxide synthase) inhibitor, an environmental contaminant, a xenobiotic and a drug allergen. It is a member of benzophenones and an oxo monocarboxylic acid. It is functionally related to a propionic acid. Ketoprofen, a propionic acid derivative, is a nonsteroidal anti-inflammatory agent (NSAIA) with analgesic and antipyretic properties. Ketoprofen is a Nonsteroidal Anti-inflammatory Drug. The mechanism of action of ketoprofen is as a Cyclooxygenase Inhibitor. Ketoprofen is a nonsteroidal antiinflammatory drug (NSAID) used in treatment of acute pain and chronic arthritis. Ketoprofen has been linked to a low rate of serum enzyme elevations during therapy and to rare instances of clinically apparent acute liver injury. Ketoprofen has been reported in Homo sapiens with data available. Ketoprofen is a propionic acid derivate and nonsteroidal anti-inflammatory drug (NSAID) with anti-inflammatory, analgesic and antipyretic effects. Ketoprofen inhibits the activity of the enzymes cyclo-oxygenase I and II, resulting in a decreased formation of precursors of prostaglandins and thromboxanes. The resulting decrease in prostaglandin synthesis, by prostaglandin synthase, is responsible for the therapeutic effects of ibuprofen. Ketoprofen also causes a decrease in the formation of thromboxane A2 synthesis, by thromboxane synthase, thereby inhibiting platelet aggregation. An IBUPROFEN-type anti-inflammatory analgesic and antipyretic. It is used in the treatment of rheumatoid arthritis and osteoarthritis. See also: Ketoprofen lysine (is active moiety of); Ketoprofen sodium (is active moiety of); Ketoprofen; Tulathromycin (component of) ... View More ... Drug Indication For symptomatic treatment of acute and chronic rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, primary dysmenorrhea and mild to moderate pain associated with musculotendinous trauma (sprains and strains), postoperative (including dental surgery) or postpartum pain. FDA Label Treatment of musculoskeletal and connective tissue pain Mechanism of Action The anti-inflammatory effects of ketoprofen are believed to be due to inhibition cylooxygenase-2 (COX-2), an enzyme involved in prostaglandin synthesis via the arachidonic acid pathway. This results in decreased levels of prostaglandins that mediate pain, fever and inflammation. Ketoprofen is a non-specific cyclooxygenase inhibitor and inhibition of COX-1 is thought to confer some of its side effects, such as GI upset and ulceration. Ketoprofen is thought to have anti-bradykinin activity, as well as lysosomal membrane-stabilizing action. Antipyretic effects may be due to action on the hypothalamus, resulting in an increased peripheral blood flow, vasodilation, and subsequent heat dissipation. Pharmacodynamics Ketoprofen is a nonsteroidal anti-inflammatory agent (NSAIA) with analgesic and antipyretic properties. Ketoprofen has pharmacologic actions similar to those of other prototypical NSAIDs, which inhibit prostaglandin synthesis. Ketoprofen is used to treat rheumatoid arthritis, osteoarthritis, dysmenorrhea, and alleviate moderate pain. 1. Ketoprofen (RP-19583) is a non-steroidal anti-inflammatory drug (NSAID) with improved COX-2 selectivity (ratio 2.9) compared to its parent analog, achieved through structure-based modification to enhance binding to the COX-2 active site [1] 2. The anti-obesity effect of ketoprofen is mediated by the mTORC1-p38-COX-2 signaling pathway: activation of mTORC1 and p38 upregulates COX-2, which further promotes the expression of white fat browning markers (UCP1, PGC-1α), increasing energy expenditure and reducing fat accumulation [2] 3. In dairy cows, ketoprofen alleviates mammary inflammation by inhibiting the TLR4-mediated pro-inflammatory cytokine (IL-6, TNF-α) secretion and promoting anti-inflammatory cytokine (IL-10) production, which has potential applications in the prevention and treatment of bovine mastitis [3] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.83 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 (9.83 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (9.83 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.9327 mL | 19.6634 mL | 39.3267 mL | |
| 5 mM | 0.7865 mL | 3.9327 mL | 7.8653 mL | |
| 10 mM | 0.3933 mL | 1.9663 mL | 3.9327 mL |