PF-06726304

CatalogusnummerS8494 Batch:S849401

Afdrukken

Technische gegevens

Formule

C₂₂H₂₁Cl₂N₃O₃

Moleculair gewicht

446.33

CAS-nr.

1616287-82-1

Oplosbaarheid (25°C)*

In vitro

Ethanol

22 mg/mL (49.29 mM)

DMSO

10 mg/mL (22.4 mM)

Water

Insoluble

In vivo (Voeg oplosmiddelen afzonderlijk en in volgorde toe aan het product.)

Homogeneous suspension

CMC-NA

≥5mg/ml

Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

* <1 mg/ml betekent licht oplosbaar of onoplosbaar.
* Houd er rekening mee dat Selleck de oplosbaarheid van alle verbindingen intern test en de werkelijke oplosbaarheid enigszins kan afwijken van gepubliceerde waarden. Dit is normaal en is te wijten aan lichte batch-tot-batch variaties.
* Verzending op kamertemperatuur (Stabiliteitstests tonen aan dat dit product zonder koelmaatregelen kan worden verzonden.)

Voorbereiden van stamoplossingen

Biologische activiteit

Beschrijving

PF-06726304 is een selectieve EZH2-remmer met Ki-waarden van respectievelijk 0,7 nM en 3 nM voor WT EZH2 en Y641N; deze verbinding remt ook H3K27me3 met een IC50-waarde van 15 nM.

Doelen

Ezh2 (wild-type) (cell-free)	EZH2 Y641N (cell-free)
0.7 nM(Ki)	3 nM(Ki)

In vitro

PF-06726304 vertoont zeer krachtige biochemische EZH2-remming, evenals goede activiteit in de H3K27Me3-reductie en antiproliferatie-effect in cellen.

In vivo

PF-06726304 vertoont robuuste in vivo antitumorale groei-activiteit en dosisafhankelijke derepressie van EZH2-doelgenen. Deze verbinding vertoont goede werkzaamheid in een diffuus grootcellig B-cellymfoom Karpas-422 tumormodel en vertoonde on-target farmacodynamische effecten in vivo.

Protocol (uit referentie)

Celassay:^{^[1]}	Cellijnen Karpas-422 cell line Concentraties -- Incubatietijd 72 h Methode Karpas-422 cells are plated in complete cell culture medium (100 μL/well) in a 96-well clear, V bottom polystyrene cell culture plate at a density of 2500 cells/well. The cells are then incubated for 2−3 h at 37 °C and 5% CO2. Compound dilution plates are prepared in 96-well clear, U-bottom polypropylene plates in 100% DMSO at 10 mM stock concentration in duplicate wells using an 11-point serial dilution (1:3 dilutions). This compound is further diluted in growth medium, and 25 μL is added to each well of the cell plates. The highest this chemical concentration tested is 50 μM final, with a 0.5% final DMSO concentration. The plates are then incubated for 72 h at 37 °C and 5% CO2. At the end of the incubation period with this compound, the plates are centrifuged at 2000 rpm for 5 min at room temperature, and the medium is removed. Then 100 μL of acid-extracted solution is added to each well. The plates are shaken for 50 min at 4 °C to lyse the cells, and then 38 μL of neutralization buffer is added. The plates are then frozen at −80 °C. The next day, the plates are shaken at room temperature to thaw. Cell lysates (50 μL/well) are then transferred to ELISA plates. The plates are covered and incubated for 2.5 h at room temperature with constant slow-speed shaking. After incubation, the plates are washed seven times (300 μL/well) with 1× wash buffer. Then 100 μL of biotinylated trimethylhistone H3K27 detection antibody is added to each well and incubated for 2 h at room temperature with constant slow-speed shaking. After incubation, the plates are washed as described above. Horseradish peroxidase-linked antibody (100 μL) is added to each well and incubated for 60 min at room temperature with constant slow-speed shaking. After incubation, the plates are washed. Finally, 100 μL of TMB substrate reagent is added to each well and incubated for 5 min at room temperature in the dark with slow shaking, after which 100 μL of stop solution is added to stop the reaction.
Dierstudie:^{^[1]}	Dierlijke modellen Female Scid beige mice Doseringen 30, 100, 300 mg/kg Toediening oral

Celassay:^{^[1]}

Cellijnen
Karpas-422 cell line
Concentraties
--
Incubatietijd
72 h
Methode
Karpas-422 cells are plated in complete cell culture medium (100 μL/well) in a 96-well clear, V bottom polystyrene cell culture plate at a density of 2500 cells/well. The cells are then incubated for 2−3 h at 37 °C and 5% CO2. Compound dilution plates are prepared in 96-well clear, U-bottom polypropylene plates in 100% DMSO at 10 mM stock concentration in duplicate wells using an 11-point serial dilution (1:3 dilutions). This compound is further diluted in growth medium, and 25 μL is added to each well of the cell plates. The highest this chemical concentration tested is 50 μM final, with a 0.5% final DMSO concentration. The plates are then incubated for 72 h at 37 °C and 5% CO2. At the end of the incubation period with this compound, the plates are centrifuged at 2000 rpm for 5 min at room temperature, and the medium is removed. Then 100 μL of acid-extracted solution is added to each well. The plates are shaken for 50 min at 4 °C to lyse the cells, and then 38 μL of neutralization buffer is added. The plates are then frozen at −80 °C. The next day, the plates are shaken at room temperature to thaw. Cell lysates (50 μL/well) are then transferred to ELISA plates. The plates are covered and incubated for 2.5 h at room temperature with constant slow-speed shaking. After incubation, the plates are washed seven times (300 μL/well) with 1× wash buffer. Then 100 μL of biotinylated trimethylhistone H3K27 detection antibody is added to each well and incubated for 2 h at room temperature with constant slow-speed shaking. After incubation, the plates are washed as described above. Horseradish peroxidase-linked antibody (100 μL) is added to each well and incubated for 60 min at room temperature with constant slow-speed shaking. After incubation, the plates are washed. Finally, 100 μL of TMB substrate reagent is added to each well and incubated for 5 min at room temperature in the dark with slow shaking, after which 100 μL of stop solution is added to stop the reaction.

Dierstudie:^{^[1]}

Dierlijke modellen
Female Scid beige mice
Doseringen
30, 100, 300 mg/kg
Toediening
oral

Referenties

http://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.6b00515

Sellecks PF-06726304 Is geciteerd door 3 Publicaties

Genetic and epigenetic characterization of sarcoma stem cells across subtypes identifies EZH2 as a therapeutic target [ NPJ Precis Oncol, 2025, 9(1):7]	PubMed: 39789291
Cathepsin-facilitated invasion of BMI1-high hepatocellular carcinoma cells drives bile duct tumor thrombi formation [ Nat Commun, 2023, 10.1038/s41467-023-42930-y]	PubMed: 37923799
Quantitative control of noise in mammalian gene expression by dynamic histone regulation [ Elife, 2021, 10e65654]	PubMed: 34379055

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VERZENDING EN OPSLAG
Selleck producten worden bij kamertemperatuur vervoerd. Als u het product op kamertemperatuur ontvangt, wees dan gerust, de Selleck kwaliteitsinspectieafdeling heeft experimenten uitgevoerd om te controleren of de normale temperatuurplaatsing van één maand de biologische activiteit van poederproducten niet beïnvloedt. Na ontvangst dient u het product op te slaan volgens de vereisten beschreven in het gegevensblad. De meeste Selleck producten zijn stabiel onder de aanbevolen omstandigheden.

NIET VOOR HUMANE, VETERINAIRE DIAGNOSTISCHE OF THERAPEUTISCHE DOELEINDEN.