Acalabrutinib acp 196

Obinutuzumab was a kind gift from Roche Glycart. Rituximab and alemtuzumab were obtained from the hospital pharmacy. Antibodies were used at a concentration of 10 μg/ml that is known to elicit maximal effects. The PI3K inhibitors idelalisib, duvelisib (IPI-145), and copanlisib (BAY 80-6946) as well as the irreversible BTK inhibitors ibrutinib, acalabrutinib (ACP-196), and tirabrutinib (ONO/GS-4059) were purchased from Selleck via AbSource (Munich, Germany) and used as stock solutions prepared in DMSO. The DMSO concentration in cell culture media was limited to 0.5%.

Human cell lines MEC-1 (CLL) and ABC-DLBCL (RIVA and TMD8) were cultured in RPMI-1640 medium supplemented with 10% FBS (Atlanta Biologicals, Lawrenceville, GA), and antibiotic-antimycotic (Gibco, Life Technologies, Gaithersburg, MD). Ibrutinib-R (IB-R) and Acalabrutinib-R (Aca-R) cells were cultured with 5% FBS. Cell lines were routinely screened for Mycoplasma, variations in growth rates, changes in morphological characteristics, and their response to stress with Annexin V FITC-PI staining; their passage number did not exceed 20. Ibrutinib was obtained from Santa Cruz (Chicago, IL); Acalabrutinib (ACP-196) from Selleck Chemicals (Houston, TX).
Peripheral blood samples were obtained from CLL patients after informed consent according to protocols approved by the institutional review board (IRB) according to the Declaration of Helsinki. Patient characteristics were as described earlier [18 ]. Lymphocytes from these blood samples were purified by Ficoll-Paque PLUS (Amersham Biosciences, Piscataway, NJ) gradient centrifugation.

Zanubrutinib (BGB-3111), ibrutinib (S2680), and acalabrutinib (ACP-196) were purchased from Selleckchem. The kinase inhibitor library was obtained from the Lerner Research Institute Molecular Screening Core. [³H]-labeled DHEA (100 nM, 300,000–600,000 cpm) was purchased from PerkinElmer, and steroids were purchased from Steraloids.