Darbufelone mesylate (CI-1004 mesylate)
(Synonyms: 甲磺酸达布非龙,CI-1004 mesylate) 目录号 : GC30604
Darbufelone mesylate (CI-1004 mesylate) (CI-1004 mesylate) 是细胞 PGF2α 的双重抑制剂；和 LTB4 生产。
Cas No.:139340-56-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Kinase experiment: | The effect of Darbufelone on the cyclooxygenase activity of PGHS-2 is determined in assays with no enzyme-inhibitor preincubation. HoloPGHS-2 (30 nM final concentration) is added to reaction mixtures that contain 20 mM Tris-HCl buffer (pH 7.4), 100 μM TMPD, and varying levels of Arachidonic acid (0-60 μM) and Darbufelone (0-30 μM). The cyclooxygenase activity is measured by monitoring the oxidation of TMPD at 610 nm using a microplate reader[1]. |
Cell experiment: | A549 (CCL-185, lung adenocarcinoma cancer cell line), NCI-H520 (HTB-182, lung squamous cancer cell line) and NCI-H460 (HTB-177, lung large cell cancer cell line) cells are cultured in RPMI-1640, supplemented with 10% (v/v) heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 U/mL Penicillin G, and 100μg/mL Streptomycin. Cells are grown at 37°C in a humidified atmosphere of 95% air and 5% CO2 and routinely passaged using 0.25% trypsin–EDTA. The effect of Darbufelone on human lung carcinoma cell viability is determined by MTT reduction assay. In brief, tumor cells growing in log-phase are trypsinized and seeded at 5×103 cells per well into 96-well plates and allowed to attach overnight. Medium in each well is replaced with fresh medium or medium containing various concentrations of Darbufelone (5-60 μM) in at least triplicate wells. Cells are cultured to another 72 h. After treatment, 1/10 volume of MTT solution (5 mg/mL) is added to each well, and the plate is incubated at 37°C for another 4 h. Two hundred microliters of DMSO is added to each well to solubilize the MTT-formazan product after removal of the medium. Absorbance at 595 nm is measured with a multi-well spectrophotometer. Growth inhibition is calculated as a percentage of the untreated controls[2]. |
Animal experiment: | Mice[2]C57Bl/6 male mice at 4-5 weeks are used. These mice are housed in air-conditioned quarters and are provided food and water ad libitum. On the day 0, Lewis Lung Carcinoma cells (1×106) are implanted into the left armpit of C57Bl/6 mice. The mice are randomly divided into four treatment groups of ten animals each. The day after inoculation (day 1), control group is treated with CMC-Na, and other groups are administered Darbufelone by gavage at doses of 20, 40, and 80 mg/kg/day. The treatment is continued till the end of the study. On day 14, animals are killed, and tumors are excised, weighed, and fixed in formalin for the further histochemical analysis. |
References: [1]. Johnson AR, et al. Slow-binding inhibition of human prostaglandin endoperoxide synthase-2 with darbufelone, an isoform-selective antiinflammatory di-tert-butyl phenol. Biochemistry. 2001 Jun 26;40(25):7736-45. | |
Darbufelone mesylate is a dual inhibitor of cellular PGF2α and LTB4 production. Darbufelone potently inhibits PGHS-2 (IC50 = 0.19 μM) but is much less potent with PGHS-1 (IC50= 20 μM).
Darbufelone is a noncompetitive inhibitor of PGHS-2 (Ki=10±5 μM). Darbufelone quenches the fluorescence of PGHS-2 at 325 nm (lambda(ex)=280 nm) with Kd=0.98±0.03 μM[1].To test the putative anti-proliferative effect of Darbufelone, A549, H520 and H460 cell lines are used, which are established from three distinct pathological subtypes of NSCLC (adenocarcinoma, squamous and large cell lung cancer respectively). Increasing concentrations of Darbufelone, ranging from 5 to 60 μM, are tested for 72 h. The cell growth inhibition of these three cell lines gradually increases with higher drug concentration. The IC50 of A549 and H520 are 20±3.6 and 21±1.8 μM, respectively, while the H460 has much lower IC50 (15±2.7 μM) [2].
Darbufelone is a dual inhibitor of cellular PGF2R and LTB4 production. Darbufelone is orally active and nonulcerogenic in animal models of inflammation and arthritis[1]. When mice are treated with Darbufelone at dosage of 80 mg/kg/day, the tumor volumes decrease in a time-dependent manner. In contrast, lower dose of Darbufelone (20 or 40 mg/kg/day) dos not show any significant inhibition of tumor weight. At necropsy, the tumor weight in mice treated with Darbufelone (80 mg/kg/day) is reduced by 30.2% in comparison with control group[2].
[1]. Johnson AR, et al. Slow-binding inhibition of human prostaglandin endoperoxide synthase-2 with darbufelone, an isoform-selective antiinflammatory di-tert-butyl phenol. Biochemistry. 2001 Jun 26;40(25):7736-45. [2]. Ye X, et al. Darbufelone, a novel anti-inflammatory drug, induces growth inhibition of lung cancer cells both in vitro and in vivo. Cancer Chemother Pharmacol. 2010 Jul;66(2):277-85.
| Cas No. | 139340-56-0 | SDF | |
| 别名 | 甲磺酸达布非龙,CI-1004 mesylate | ||
| Canonical SMILES | O=C1N=C(N)S/C1=C\C2=CC(C(C)(C)C)=C(O)C(C(C)(C)C)=C2.CS(=O)(O)=O | ||
| 分子式 | C19H28N2O5S2 | 分子量 | 428.57 |
| 溶解度 | DMSO : 110 mg/mL (256.67 mM; Need ultrasonic); H2O : < 0.1 mg/mL (insoluble) | 储存条件 | 4°C, protect from light |
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.3333 mL | 11.6667 mL | 23.3334 mL |
| 5 mM | 0.4667 mL | 2.3333 mL | 4.6667 mL |
| 10 mM | 0.2333 mL | 1.1667 mL | 2.3333 mL |
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Effect of arachidonic acid cascade inhibitors on body temperature and cognitive functions in rats in the Morris water maze after acute cold injury
Objective: The aim of the study was to evaluate the effect of arachidonic acid cascade inhibitors on body temperature and cognitive functions of rats (spatial memory, learning ability) in the Morris water maze test (MWM) after acute cold injury (CI). Methods: Animals were trained to find an escape platform in the MWM for two consecutive days. On the third day, rats were treated with saline (10 ml/kg), diclofenac sodium (7 mg/kg), etoricoxib (5 mg/kg), darbufelone mesylate (20 mg/kg) or montelukast (1 mg/kg) intragastrically (i.g.), 30 minutes before CI modeling. Air hypothermia with an acute general cooling (AGC) model was used as a kind of CI. Animals were subjected to cooling for 2 hours at -18°C. Body temperature was measured before and after CI modeling. MWM experimental trials tests were carried out after cold exposure. Number of successful trials, escape latency, distance moved, velocity, meander, and behavioral patterns in individual quadrants were recorded. Results: In the control pathology group, a statistically significantly body temperature decrease was observed (p<0.05 compared to the initial value). All of the studied drugs reduced hypothermia severity, but only in the sodium diclofenac group this reduction reached a significant level in comparison with the untreated animals (p<0.01). A tendency to reduce the severity of hypothermia was observed in the group of animals treated by etoricoxib, darbufelone mesylate, and montelukast. In the control pathology group, the number of successful trials was significantly decreased (p<0.01), velocity (p<0.05), and escape latency (p<0.05) were increased compared with intact animals. Diclofenac sodium significantly reduced escape latency (p<0.05) and increased the number of successful trials in comparison with the control pathology group (p<0.01). Montelukast tended to improve, etoricoxib and darbufelone mesylate did not improve cognitive functions of rats with CI. Conclusions: The results experimentally substantiate the possibility of effective pharmacoprophylaxis of CI and its negative effects on cognitive functions while applying arachidonic acid cascade inhibitors, particularly the non-selective COX inhibitor diclofenac sodium.
Synthesis and biological evaluation of 3-[4-(amino/methylsulfonyl)phenyl]methylene-indolin-2-one derivatives as novel COX-1/2 and 5-LOX inhibitors
Fourteen new 3-[4-(amino/methylsulfonyl)phenyl]methylene-indolin-2-one derivatives were synthesized. Six compounds displayed potent inhibitory activities against COX-1/2 and 5-LOX with IC(50) in the range of 0.10-9.87 μM. Particularly, 10f exhibited well balanced inhibitory action on these enzymes (IC(50)=0.10-0.56 μM). More importantly, 10f and several other compounds had comparable or stronger anti-inflammatory and analgesic activities, but better gastric tolerability in vivo, as compared with darbufelone mesilate and tenidap sodium. Therefore, our findings may aid in the design of new and safe anti-inflammatory reagents for the intervention of painful inflammatory diseases, such as rheumatoid arthritis at clinic.
Slow-binding inhibition of human prostaglandin endoperoxide synthase-2 with darbufelone, an isoform-selective antiinflammatory di-tert-butyl phenol
The antiinflammatory agent darbufelone, ((Z)-5-[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl] methylene]-2-imino-4-thiazolidinone, methanesulfonate salt), was discovered as a dual inhibitor of cellular prostaglandin and leukotriene production. To study the mechanism of action of this drug, we expressed human prostaglandin endoperoxide synthase-1 (PGHS-1) and PGHS-2 and purified the recombinant enzymes using buffers that contain octylglucoside. In cyclooxygenase assays following a 15-min incubation of enzyme with inhibitor, darbufelone potently inhibits PGHS-2 (IC(50) = 0.19 microM) but is much less potent with PGHS-1 (IC(50) = 20 microM). Interestingly, when the assay buffer contains traces of Tween 20 (0.0001%), darbufelone appears inactive with PGHS-2 due to a detergent interaction that is detectable by absorption spectroscopy. We therefore used octylglucoside, which does not affect darbufelone in this way, in place of Tween 20 in our PGHS buffers. Inhibition of PGHS-2 with darbufelone is time dependent: with no preincubation, darbufelone is a weak inhibitor (IC(50) = 14 microM), but after a 30-min incubation it is 20-fold more potent. Plots of PGHS-2 activity vs preincubation time at various darbufelone concentrations reach a plateau. This finding is inconsistent with irreversible or one-step slow-binding inhibition. A two-step slow-binding inhibition model is proposed in which the E.I complex (K(i) = 6.2 +/- 1.9 to 14 +/- 1 microM) slowly transforms (k(5) = 0.015-0.030 s(-)(1)) to a tightly bound E.I form with K(i) = 0.63 +/- 0.07 microM and k(6) = 0.0034 s(-)(1). In steady-state kinetics inhibition experiments performed with no preincubation, we find that darbufelone is a noncompetitive inhibitor of PGHS-2 (K(i) = 10 +/- 5 microM). Darbufelone quenches the fluorescence of PGHS-2 at 325 nm (lambda(ex) = 280 nm) with K(d) = 0.98 +/- 0.03 microM. The PGHS substrate, arachidonate, and various cyclooxygenase inhibitors do not alter this binding affinity of darbufelone but a structural analogue of darbufelone competes directly for binding to PGHS-2. Di-tert-butyl phenols such as darbufelone may inhibit PGHS-2 by exploiting a previously unrecognized binding site on the enzyme.