human COX-1 (IC50 = 18 nM in CHO cells); hCOX-2 (IC50 = 26 nM in CHO cells)

In vitro antitumor activity of indomethacin sodium hydrate (0-150 μM; 24 hours; 3LL-D122 cells) has been shown [2]. By activating PKR and phosphorylating eLF2α, indomethacin sodium hydrate (0-1000 μM) inhibits viral replication (IC50=2 μM) and stops viral protein translation, protecting host cells from viral harm [3]. NaCl indomethacin hydrate

Carrageenan-induced hyperalgesia is dose-dependently reversed by indomethacin (Indometacin) sodium hydrate (0.01-10 mg/kg; oral; 3 hours; male Sprague-Dawley rats) which also causes paw edema and hyperalgesia [1]. Tumor growth is inhibited in vivo by indomethacin (10 mg/mL; oral; once daily for 29 days; male C57BL/6J mice) [2].

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Using indomethacin (IND), gastric ulcer model can be generated in animals as detailed below:
Generation of Gastric Ulcer Model: all animals fasted 24 h before drug administration. Except for the control group, ulcers were induced by administering IND to the three experimental study groups, namely IND, IND+ ESP and IND + CA. The same volume of physiological saline was administered to the experimental animals as to the control group. 50 mg/kg ketamine and 5 mg/kg xylazine were administered to rats 6 h after IND administration. Anesthetized rats were euthanized by cervical dislocation, after which tissue samples were collected. Specifically, the stomach was opened along the greater curvature and washed with physiological saline at 4 °C. Washed stomach tissues were stored in tubes containing 10% formalin for histological procedures and at −800 °C for biochemical determination until analyses. Hematoxylin-eosin staining of the taken tissues was evaluated histopathologically and immunohistochemically.[4]

Determination of Ki and k2 values for the time-dependent inhibition of COX-2[1]
Purified COX-2 (2.3 μg) was preincubated with inhibitor for 0–15 min in 180 μl of the reaction buffer described above, before the initiation of the reaction with a mixture of arachidonic acid and TMPD. The cyclo-oxygenase activity was determined by the spectrophotometric method as described above. For experiments performed without preincubation of the inhibitor, the reaction was initiated by addition of the assay mixture containing the enzyme to the inhibitor and arachidonic acid/TMPD ethanolic solution. The rate constants (kobs) for the time-dependent loss of activity at each inhibitor concentration were calculated by fitting of the data to a first order equation of the form y=a + b.exp(–kobst) by use of Sigmaplot software. Data were analysed in terms of the model developed by Rome and Lands (1975) for the time-dependent inhibition of ovine COX-1. In this model (Scheme 1), an initial reversible binding of enzyme and inhibitor (characterized by the dissociation constant Ki) is followed by a first order inactivation process (characterized by a first order rate constant k2). The rate of reversal of this process (k–2) is considered to be negligible.

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Determination of the stoichiometry of inhibitor binding[1]
Aliquots of purified COX-2 (0.25 mg ml-1, concentration of subunit of 3.4 μm) were incubated in buffer (100 mm Tris-HCl, pH 8.0, 5 mm EDTA, 1 mm phenol) in the presence of varying concentrations of inhibitors (0–8 μm) for 15 or 30 min. An aliquot (20 μl) was then removed for determination of the remaining cyclo-oxygenase activity by oxygen uptake as described above. Enzyme concentration was determined by amino acid concentration following acid hydrolysis (Percival et al., 1994).

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Competition of time-dependent inhibition of COX-2 by arachidonic acid[1]
Purified COX-2 (3.6 μg) was diluted into preincubation buffer (0.03 ml, 100 mm Tris-HCl, pH 8.0, 5 mm EDTA, 2 mm phenol) containing 60 mm diethyldithiocarbamic acid to prevent substrate oxygenation (Lands et al., 1974) and either 10 μm inhibitor, or 10 μm inhibitor plus 5 μm arachidonic acid, or 10 μm inhibitor plus 30 μm arachidonic acid. After a preincubation period of 0–4 min, the total enzyme was assayed for enzymatic activity by oxygen consumption at 30°C as described above.

Cell Viability Assay[2]
Cell Types: 3LL-D122 cells (highly metastatic variant of mouse LLcarcinoma cells)
Tested Concentrations: 0, 20, 50, 100 and 150μM
Incubation Duration: 24 hrs (hours)
Experimental Results: Inhibited cell viability at 20 mM, with 50% inhibition at 60 mM.

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Cell Cycle Analysis[2]
Cell Types: 3LL-D122 cells (highly metastatic variant of mouse LLcarcinoma cells)
Tested Concentrations: 0, 30 and 80μM
Incubation Duration: 24 hrs (hours)
Experimental Results: diminished in the percentage of cells at the G2/M phase and increased in the percentage of cells at G1 phase.

Animal/Disease Models: Male SD (Sprague-Dawley) rats[1]
Doses: 0.01-10 mg/kg
Route of Administration: Oral administration; for 3 hrs (hours)
Experimental Results: Inhibited the carrageenan- induced rat paw oedema (ED50=2.0 mg/kg) and hyperalgesia (ED50=1.5 mg/kg) in a dose-dependent manner.

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Animal/Disease Models: Male C57BL/6J mice[2]
Doses: 10 mg/mL
Route of Administration: Oral administration ; daily, for 29 days
Experimental Results: Delayed the onset of tumor growth and the initial growth rate of the footpad tumors.

[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]. Comparative effects of indomethacin on cell proliferation and cell cycle progression in tumor cells grown in vitro and in vivo. Biochem Pharmacol. 2001 Mar 1;61(5):565-71.

[3]. Inhibition of viral protein translation by indomethacin in vesicular stomatitis virus infection: role of eIF2α kinase PKR. Cell Microbiol. 2015 Sep;17(9):1391-404.

Indomethacin Sodium is the sodium salt of indomethacin, a methylated indole derivative with anti-inflammatory, analgesic-antipyretic and tocolytic effects. Indomethacin is a non-selective, reversible, and competitive inhibitor of cyclooxygenases 1 and 2, thereby blocking the conversion of arachidonic acid into prostaglandin precursors. Consequently, prostaglandin synthesis is decreased, and prostaglandin-mediated activities are prevented, including pain, inflammation, fever and uterine contraction.
See also: Indomethacin Sodium (annotation moved to).

C19H21CLNNAO7

433.82

357.076

C, 52.60; H, 4.88; Cl, 8.17; N, 3.23; Na, 5.30; O, 25.82

74252-25-8

Indomethacin;53-86-1;Indomethacin-d4;87377-08-0;Indomethacin;53-86-1;Indomethacin sodium;7681-54-1; Indomethacin sodium hydrate;74252-25-8; 7681-54-1 (sodium)

23674731

Light yellow to yellow solid powder

1.3±0.1 g/cm3

499.4±45.0 °C at 760 mmHg

162ºC

255.8±28.7 °C

0.0±1.3 mmHg at 25°C

1.619

3.11

3

7

4

29

512

0

UHYAQBLOGVNWNT-UHFFFAOYSA-M

InChI=1S/C19H16ClNO4.Na.3H2O/c1-11-15(10-18(22)23)16-9-14(25-2)7-8-17(16)21(11)19(24)12-3-5-13(20)6-4-12;;;;/h3-9H,10H2,1-2H3,(H,22,23);;3*1H2/q;+1;;;/p-1

sodium;2-[1-(4-chlorobenzoyl)-5-methoxy-2-methylindol-3-yl]acetate;trihydrate

Indomethacin sodium trihydrate; 74252-25-8; Indomethacin sodium salt trihydrate; Indometacin sodium; Indomethacin sodium hydrate; 0IMX38M2GG; Indometacin (sodium hydrate); Indometacin sodium trihydrate;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : 25 mg/mL (57.63 mM)
DMSO : 12.5 mg/mL (28.81 mM )

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.79 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 20.8 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.79 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 20.8 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.

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Solubility in Formulation 3: 5 mg/mL (11.53 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).

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 (Please use freshly prepared in vivo formulations for optimal results.)