COX-1 (IC50 = 28 nM)

The anti-platelet actions of 1-[(4,5-bis(4-methoxyphenyl)-2- thiazoyl)carbonyl]-4-methylpiperazine hydrochloride (FR122047) were investigated in vitro and in vivo. FR122047 was 100 times more potent than aspirin against arachidonic acid- and collagen-induced human and guinea-pig platelet aggregation in vitro. Its actions on platelets were a result of cyclooxygenase inhibition[1].

The single oral dose of FR122047 inhibited arachidonic acid- and collagen-induced aggregation with an ED50 of 280 micrograms/kg and 530 micrograms/kg, respectively, in guinea-pigs. The anti-platelet action was augmented 5-10 times by repeated administration for 4 days. At 1 mg/kg the inhibitory actions were prolonged for 48 h and the drug concentration was < 0.1 ng/ml in platelet-poor plasma at 24 h and 0.282 ng/ml in platelet-rich plasma at 48 h. The safety margin in rats (minimum ulcerogenic dose/ED50 for anti-platelet aggregation) of FR122047 was more than 70, while that of aspirin was only 1.2. These results indicate that FR122047 is concentrated in platelets and may be a useful anti-platelet agent.[1]

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Oral administration of FR122047 (3.2-100 mg/kg) dose dependently reduced the phase 2 response (10-60 min) of the formalin test in rats. This effect was 3 times less potent than that of indomethacin. FR122047 (1-32 mg/kg; p. o.) showed a dose-dependent analgesic effect against the acetic acid-induced writhing response in mice. Furthermore, FR122047 (0. 01-10 mg/kg, p.o.) inhibited the increase in 6-keto prostaglandin F(1alpha) level in acetic acid-injected mouse peritoneal cavity. However, a selective cyclooxygenase-2 inhibitor, NS-398, had no effect in these cyclooxygenase-1 sensitive pain models. These results suggest that FR122047, a selective cyclooxygenase-1 inhibitor, shows an analgesic effect in chemical nociceptive models and may be a useful analgesic agent[2].

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In this study, researchers investigated the effects of FR122047 (1-[(4,5-bis(4-methoxyphenyl)-2-thiazoyl)carbonyl]-4-methylpiperazine hydrochloride), a selective cyclo-oxygenase (COX)-1 inhibitor, in rat type II collagen-induced arthritis (CIA) and adjuvant-induced arthritis (AIA). Using an ex vivo rat whole blood assay, FR122047 (0.032 - 3.2 mg kg(-1)) inhibited COX-1-derived thromboxane (TX) B(2) production with ED(50) value of 0.059 mg kg(-1), indicating that it was orally active, but did not inhibit lipopolysaccharide-induced prostaglandin (PG) E(2) production derived by COX-2. Oral administration of FR122047 showed a dose-dependent anti-inflammatory effect in rat CIA with ED(50) value of 0.56 mg kg(-1). This drug also dose dependently suppressed the levels of PGE(2) and TXB(2) in CIA rat paws with ED(50) values of 0.24 and 0.13 mg kg(-1), respectively. FR122047 had no effect in rat AIA model. In contrast, indomethacin, a non-selective COX inhibitor, was anti-inflammatory and reduced the formation of PGs in AIA rat paws. Unlike indomethacin, chronic treatment of FR122047 did not damage the stomach mucosa in CIA rats. These results demonstrate that COX-1 contributes to the oedema and the formation of PGE(2) and TXB(2) in rat CIA model, but not in rat AIA model. We conclude that FR122047 has an orally active and anti-inflammatory effect mediated by inhibition of PGE(2) and TXB(2) produced by COX-1 at a site of inflammation induced by type II collagen and it may be a useful tool for studying the involvement of COX-1 in various in vivo models of inflammation[3].

The pharmacological profile of the analgesic agent, 1-[(4, 5-bis(4-methoxyphenyl)-2-thiazoyl)carbonyl]-4-methylpiperazine hydrochloride (FR122047), was investigated. In recombinant human cyclooxygenase enzyme assays, the inhibition of prostaglandin E(2) formation by FR122047 was 2300 times more selective for cyclooxygenase-1 than cyclooxygenase-2[2].

In COX-1 assay, blood was collected in siliconized tubes containing no anticoagulants. Aliquots of 500 μl were immediately transferred to siliconized microcentrifuge tubes. The tubes were vortexed and incubated at 37°C for 1 h to allow the blood to clot. Reactions were terminated by the addition of 5 μl of indomethacin 1 mm, and serum was obtained by centrifugation at 12,000× g for 5 min at 4°C. A 100 μl aliquot of serum was mixed with 400 μl methanol for protein precipitation. The supernatant was obtained and was assayed for thromboxane (TX) B2 by radioimmunoassay.[3]

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In COX-2 assay, blood was collected in heparinized tubes. Aliquots of 500 μl blood were immediately transferred to siliconized microcentrifuge tubes, and were incubated for 15 min at 37°C. This was followed by incubation of the blood with 10 μl lipopolysaccharide (LPS) ( #L-2630 from E. coli serotype 0111:B4, 100 μg ml−1 final concentration, in phosphate-buffered saline) for 24 h at 37°C for induction of COX-2. Reactions were terminated by the addition of 5 μl of indomethacin 1 mm, and the blood was centrifuged at 12,000× g for 5 min at 4°C to obtain plasma. A 100 μl aliquot of plasma was mixed with 400 μl methanol for protein precipitation. The supernatant was obtained and assayed for PGE2 by radioimmunoassay[3]

The technique of Brideau et al. (1996) was modified. Single oral doses of 0.032 – 3.2 mg kg−1 of drugs were administered to rats. Blood samples were taken at 1.5 h after dosing. FR122047 was suspended and diluted in 0.5% methylcellulose.[3]

Induction of type II collagen-induced arthritis[3]
Type II collagen (CII) isolated and purified from bovine articular cartilage was purchased from Collagen Research Center (Tokyo, Japan) and dissolved overnight at 4°C in 0.01 m acetic acid at a concentration of 2 mg ml−1. The solution was emulsified in an equal volume of incomplete Freund's adjuvant. Each rat was immunized with 0.5 ml of the cold emulsion (0.5 mg CII) by several intradermal injections on the back and one or two injections into the base of the tail (Inamura et al., 1988). They were challenged with 0.2 ml of the emulsion (0.2 mg CII) into the base of the tail on day 7 after immunization. The drugs were given orally once a day prophylactically from day 1 to day 24 after the first CII immunization. For the time course study, paw volume was measured before and 7, 10, 14, 18, 21 and 24 days after the first immunization with the Volume Meter TK-105 (Neuroscience, Tokyo, Japan), and oedema was expressed as the increase in paw volume (Δ ml) after CII immunization relative to the pre-immunization value for each animal. The anti-inflammatory effect was expressed as the difference in paw oedema compared with that of vehicle-treated CIA-control rats.[3]

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Induction of adjuvant-induced arthritis[3]
A suspension of heat-killed and dried Mycobacterium tuberculosis H37 RA (0.5 mg; DIFCO) in 0.05 ml liquid paraffin was administered by intradermal injection into the plantar surface of the right hind paw at day 0 to induce adjuvant arthritis (Newbould, 1963; Walz et al., 1971). The drugs were given orally once a day prophylactically from day 1 to day 24. Paw volume was measured before and 24 days after adjuvant injection with the Volume Meter TK-105, and the anti-inflammatory effect was expressed as the difference in paw oedema compared with that of vehicle-treated AIA-control rats.[3]

[1]. The anti-platelet actions of FR122047, a novel cyclooxygenase inhibitor. Eur J Pharmacol. 1993 Oct 19;243(2):179-84.

[2]. Ochi T, Motoyama Y, Goto T. The analgesic effect profile of FR122047, a selective cyclooxygenase-1 inhibitor, in chemical nociceptive models. Eur J Pharmacol. 2000 Mar 10;391(1-2):49-54.

[3]. Ochi T, Goto T. Differential effect of FR122047, a selective cyclo-oxygenase-1 inhibitor, in rat chronic models of arthritis. Br J Pharmacol. 2002 Feb;135(3):782-8.

See also: FR122047 (annotation moved to).

C23H26CLN3O3S

459.989

459.138

C, 60.06; H, 5.70; Cl, 7.71; N, 9.14; O, 10.43; S, 6.97

130717-51-0

196840

White to light yellow solid powder

562.7ºC at 760 mmHg

294.1ºC

1.1E-12mmHg at 25°C

4.559

1

6

5

31

556

0

Cl.COC1C=CC(C2=C(C3C=CC(OC)=CC=3)SC(C(N3CCN(C)CC3)=O)=N2)=CC=1

YWMAVHIKOAOSFM-UHFFFAOYSA-N

InChI=1S/C23H25N3O3S.ClH/c1-25-12-14-26(15-13-25)23(27)22-24-20(16-4-8-18(28-2)9-5-16)21(30-22)17-6-10-19(29-3)11-7-17;/h4-11H,12-15H2,1-3H3;1H

[4,5-bis(4-methoxyphenyl)-1,3-thiazol-2-yl]-(4-methylpiperazin-1-yl)methanone;hydrochloride

FR-122047; FR122047; R 122047 hydrochloride; 130717-51-0; (4,5-Bis(4-methoxyphenyl)thiazol-2-yl)(4-methylpiperazin-1-yl)methanone hydrochloride; 1-[[4,5-BIS(4-METHOXYPHENYL)-2-THIAZOLYL]CARBONYL]-4-METHYLPIPERAZINE HYDROCHLORIDE; FR-122047 HCl; Methanone, [4,5-bis(4-methoxyphenyl)-2-thiazolyl](4-methyl-1-piperazinyl)-, hydrochloride (1:1); FR 122047

2934.99.9001

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.

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

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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