AZD6482

2 years -80 in solvent

Human platelet pellet

0–60 nM, dissolved in DMSO

AZD6482 also inhibits PI3Kalpha, gamma, and delta, with IC50 of 80 nM to 1.4 uM, which are significantly lower than its (+)-enantiomer (S-form). AZD6482 is an antiplatelet agent, it blocks platelet activation adhesion/aggregation and promotes platelet disaggregation in assay of washed platelet aggregation (WPA), with an IC50 value of 6 nM. Furthermore, by targeting PI3Kbeta, AZD6482 specifically inhibits thrombosis without interfering with normal haemostasis. Therefore, AZD6482 is used as an anti-thrombotic drug for the prophylaxis of thrombotic disorders. [1]

Assay of PI3K enzyme inhibition, The inhibition of PI3Kbeta, PI3Kalpha, PI3Kgamma, and PI3Kdelta is evaluated in an AlphaScreen based enzyme activity assay using human recombinant enzymes. The assay measures PI3K-mediated conversion of PIP2 to PIP3. Biotinylated PIP3, a GST-tagged pleckstrin homology (PH) domain and the two AlphaScreen beads form a complex that elicits a signal upon laser excitation at 680 nm. The PIP3 formed in the enzyme reaction competes with the biotinylated PIP3 for binding to the PH domain thus reducing the signal with increasing enzyme product., The AZD6482 is dissolved in DMSO and added to 384 well plates. PBKbeta, PBKalpha, PBKgamma, or PBKdelta is added in a Tris buffer (50 mM Tris pH 7.6, 0.05% CHAPS, 5 mM DTT, and 24 mM MgCl2) and allowed to preincubate with AZD6482 for 20 minutes prior to the addition of substrate solution containing PIP2 and ATP. The enzyme reaction is stopped after 20 minutes by addition of stop solution containing EDTA and biotin-PIP3, followed by addition of detection solution containing GST-grpl PH and AlphaScreen beads. Plates are left for a minimum of 5 hours in the dark prior to analysis. The final concentration of DMSO, ATP and PIP2 in the assay are, 0.8%, 4 uM, and 40 uM, respectively. IC50 values are calculated according to the equation, y = {a+[(b-a)/(l+(x/IC50)s)]}, where y = % inhibition, a = 0%, b = 100%, s = the slope of the concentration-response curve, x = AZD6482 concentration.

DMSO 82 mg/mL, Water <1 mg/mL, Ethanol 28 mg/mL

(R)-2-(1-(7-methyl-2-morpholino-4-oxo-4H-pyrido[1, 2-a]pyrimidin-9-yl)ethylamino)benzoic acid