AT-56

Catalog No.S9678 Batch:S967801

Technical Data

Formula	C₂₅H₂₇N₅
Molecular Weight	397.52	CAS No.	162640-98-4
Solubility (25°C)*	In vitro	DMSO	80 mg/mL (201.24 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

AT-56 is an orally active and selective inhibitor of lipocalin-type prostaglandin (PG) D synthase (L-PGDS) with Ki of 75 μM and IC50 of 95 μM.

Targets

L-PGDS ^[1] (Cell-free assay)	L-PGDS ^[1] (Cell-free assay)
75 μM(Ki)	95 μM

In vitro

AT-56 inhibits human and mouse L-PGDSs in a concentration (3–250 μm)-dependent manner. AT-56 inhibits the L-PGDS activity in a competitive manner against the substrate PGH2 (Km = 14 μm) with a Ki value of 75 μm. AT-56 inhibits the production of PGD2 by L-PGDS-expressing human TE-671 cells after stimulation with Ca2+ ionophore (5 μm A23187) with an IC50 value of about 3 μm without affecting their production of PGE2 and PGF2α.^[1]

In vivo

Orally administered AT-56 (<30 mg/kg body weight) decreases the PGD2 production to 40% in the brain of H-PGDS-deficient mice after a stab wound injury in a dose-dependent manner without affecting the production of PGE2 and PGF2α and also suppresses the accumulation of eosinophils and monocytes in the bronco-alveolar lavage fluid from the antigen-induced lung inflammation model of human L-PGDS-transgenic mice.^[1]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines TE-671 cells, MEG-01S cells Concentrations 0 - 100 μM Incubation Time 10 min Method TE-671 (1×10⁶ cells/well) and MEG-01S (5×10⁵ cells/well) are seeded into multiplates and cultured in Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum, 4 mM L-glutamine, 4.5 g/liter glucose, 100 units/ml penicillin, and 100 μg/ml streptomycin sulfate under a 5% CO2 atmosphere at 37 ℃. MEG-01S cells are caused to differentiate by treatment with 12-O-tetradecanoylphorbol-13-acetate to express H-PGDS and COX-1. After the cells have been cultured for 1 day, AT-56 is added to them at different doses ranging from 0 to 100 μM, and the cells are then incubated at 37 ℃ for 10 min. Thereafter, they are stimulated with a calcium ionophore, A23187 (5 μM), at 37 ℃ for 15 min. The culture media were harvested and centrifuged at 10,000 × g for 5 min to remove the cells, and the supernatant is removed and stored at 80 ℃ until the measurements of PGs can be made.
Animal Study:^{^[1]}	Animal Models H-PGDS KO mice (14–16-week-old males or females weighing 25–30 g, C57BL/6 strain) Dosages 1 mg/kg, 3 mg/kg, 10 mg/kg, 30 mg/kg Administration Oral gavage

Cell Assay:^{^[1]}

Cell lines
TE-671 cells, MEG-01S cells
Concentrations
0 - 100 μM
Incubation Time
10 min
Method
TE-671 (1×10⁶ cells/well) and MEG-01S (5×10⁵ cells/well) are seeded into multiplates and cultured in Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum, 4 mM L-glutamine, 4.5 g/liter glucose, 100 units/ml penicillin, and 100 μg/ml streptomycin sulfate under a 5% CO2 atmosphere at 37 ℃. MEG-01S cells are caused to differentiate by treatment with 12-O-tetradecanoylphorbol-13-acetate to express H-PGDS and COX-1. After the cells have been cultured for 1 day, AT-56 is added to them at different doses ranging from 0 to 100 μM, and the cells are then incubated at 37 ℃ for 10 min. Thereafter, they are stimulated with a calcium ionophore, A23187 (5 μM), at 37 ℃ for 15 min. The culture media were harvested and centrifuged at 10,000 × g for 5 min to remove the cells, and the supernatant is removed and stored at 80 ℃ until the measurements of PGs can be made.

Animal Study:^{^[1]}

Animal Models
H-PGDS KO mice (14–16-week-old males or females weighing 25–30 g, C57BL/6 strain)
Dosages
1 mg/kg, 3 mg/kg, 10 mg/kg, 30 mg/kg
Administration
Oral gavage

References

[1] Daisuke Irikura, et al. J Biol Chem. 2009 Mar 20;284(12):7623-30.

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