Molecular Weight (MW)

364.4

Formula

C15H17FN6O2S

CAS No.

1159824-67-5

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 5 mg/mL warming (13.72 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In vivo)

0.5% CMC: 30 mg/mL

Chemical Name

5-(2-amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide; CZC-24832; CZC24832; CZC 24832

Kinase Assay: Competition binding assays using the LK matrix are performed essentially as described above but adapted to a 384-well format. 0.25 mg of cell lysate and 2.5 μL of beads are used per well. Compounds from the screening library including reference compounds as standards are added at 40 μM final concentration from 50× DMSO stocks. Each plate contains 15 positive and 17 negative controls. After 2 hrs binding at 4 °C, the non-bound fraction is removed by washing the beads with lysis buffer. Proteins retained on the beads are eluted in SDS sample buffer and spotted on nitrocellulose membranes (400 nL/spot) using an automated pin tool liquid transfer. After drying, the membranes are rehydrated in 20% ethanol, and processed for detection with specific antibodies as indicated, followed by incubation with a labeled secondary antibody for visualization. Spot intensities are quantified using a scanner and percentage inhibition is calculated using positive and negative controls as 100% and 0% inhibition, respectively.

Cell Assay: CZC24832 has excellent selectivity to PI3Kγ. Out of the 154 identified lipid and protein kinases and the 922 other proteins, only two off targets (PI3Kβ and PIP4K2C) are detected within a 100-fold selectivity window. Despite the high sequence conservation of the human and rodent class I PI3K isoforms, the potency of CZC24832 for PI3Kγ and PI3Kβ is consistently lower by a factor of 2 to 4 in mice and rats compared to humans, but selectivity windows are largely retained. In the BT system, treatment with CZC24832 results in profound inhibition of IL-17A (IC50 =1.5 μM) as well as of B-cell activation markers such as IL-6 and IgG. In addition, strong inhibition of IL17A production is observed in T-cell systems such as human umbilical vein endothelial cells grown in the presence of TH2 blasts, indicating a general role for PI3Kγ kinase activity in the control of TH17 function. Thus, CZC24832 inhibits TH17 cell differentiation

In an IL-8–dependent air pouch model, CZC24832 shows a dose dependent reduction of granulocyte recruitment consistent with the degree of inhibition observed in PI3Kγ-null mice. In a therapeutic collagen induced arthritis (CIA) model, mice treated orally with 10 mg CZC24832 per kg body weight twice per day shows a substantial decrease of bone and cartilage destruction as well as of overall clinical parameters.

Animal model

CIA mouse model

Formulation & Dosage

Dissolved in 0.5% CMC in water; 3, 10 mg/kg; Oral gavage

References

[[1] Bergamini G, et al. Nat Chem Biol, 2012, 8(6), 576-582.

Design and characterization of CZC24832, a selective PI3Kγ inhibitor. Nat Chem Biol, 2012, 8(6), 576-582.

Cellular activity profile of CZC24832