PCI-32765

• Catalog No.: S2680
• CAS Number: 936563-96-1
• M. F.: C25H24N6O2
• M. W.: 440.49706
• Storage: -20°C

SYNONYMS

• IUPAC name: 1-{3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl}prop-2-en-1-one
• IUPAC Traditional name: 1-{3-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl}prop-2-en-1-one

DATABASE IDS

• CAS Number: 936563-96-1

PROPERTIES

• Salt Data: Free Base
• Storage Condition: -20°C

DETAILS

Description (English)

Research Area

• Description: Mantle cell lymphoma ,Chronic lymphocytic leukaemia

Biological Activity

• Description: PCI-32765 (Ibrutinib) is a potent and highly selective Btk inhibitor with IC50 of 0.5 nM.
• Targets:

  Btk

• IC50:

  0.5 nM ^[1]

• In Vitro: PCI-32765 shows the potent and irreversible inhibitory effect and selectivity for Btk enzymatic activity. In BCR pathway-activated DOHH2 cell line, PCI-32765 inhibits autophosphorylation of Btk, phosphorylation of Btk's physiological substrate PLCγ, and phosphorylation of further downstream kinase, ERK with IC50 of 11 nM, 29 nM and 13 nM, respectively. ^[1] PCI-32765 exhibits a significant dose-dependent and time-dependent induction of cytotoxicity in chronic lymphocytic leukemia (CLL) cells. In addition, PCI-32765 induces cell death depending on caspase pathway activation and antagonizes the ability of CLL cells to proliferate after TLR signaling. ^[2] A recent study shows that PCI-32765 inhibits BCR-activated primary B cell proliferation with IC50 of 8 nM and results in inhibition of TNFα, IL-1β and IL-6 production in primary monocytes with IC50 of 2.6 nM, 0.5 nM and 3.9 nM, respectively. ^[3]
• In Vivo: In a collagen-induced arthritis model, PCI-32765 significantly reduces clinical arthritis scores reflecting paw swelling and joint inflammation by inhibiting B-cell activation. In a MRL-Fas(lpr) lupus model, PCI-32765 reduces renal disease and autoantibody production. ^[1] In an adoptive transfer TCL1 mouse model of CLL, PCI-32765 (25 mg/kg/day) causes a transient early lymphocytosis, and delays CLL disease progression. ^[4]
• Clinical Trials: PCI-32765 is currently in Phase II clinical trials in patients with Chronic Lymphocytic Leukemia (CLL) or Small Lymphocytic Lymphoma (SLL). Combination of PCI-32765 and Rituximab is currently in Phase II clinical trials in patients with High-Risk Chronic Lymphocytic Leukemia (CLL) and Small Lymphocytic Lymphoma (SLL).

Combination Therapy

• Description: The use of PCI-32765 in combination with Bendamustine and Rituximab is currently in Phase III clinical trials in patients with Chronic Lymphocytic Leukemia and Small Lymphocytic Lymphoma.

Protocol

• Protocol: Kinase Assay ^[1]
• Kinase Assays: In vitro kinase IC50 values are measured using ^33P filtration binding assay after 1 hour incubation of kinase, ^33P-ATP, PCI-32765, and substrate [0.2 mg/mL poly(EY)(4:1]. Assays are performed at Reaction Biology.
• Protocol: Cell Assay ^[2]
• Cell Lines: Chronic lymphocytic leukemia (CLL) cells
• Concentrations: 0.01-100 μM
• Incubation Time: 48 hours
• Methods:

  MTT (3'[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assays are performed to determine cytotoxicity. Briefly, cells (CLL B cells or healthy volunteer T cells or NK cells) are incubated for 48 hours with different concentrations of PCI-32765, or vehicle control. MTT reagent is then added, and plates are incubated for an additional 20 hours before washing with protamine sulfate in phosphate-buffered saline. Dimethyl sulfoxide is added, and absorbance is measured by spectrophotometry at 540 nm in a Labsystems plate reader. Cell viability is also measured at various time points with the use of annexin/PI flow cytometry. Data are analyzed with Expo-ADC32 software package. Results are expressed as the percentage of total positive cells over untreated control. Experiments examining caspase-dependent apoptosis includes the addition of 100μM Z-VAD.

• Protocol: Animal Study ^[1]
• Animal Models: MRL-Fas(lpr) lupus model and collagen-induced arthritis model.
• Formulation: PCI-32765 is dissolved in DMSO.
• Doses: ≤50 mg/kg
• Administration: Administered via p.o.

References

• References: [1] Honigberg LA, et al. Proc Natl Acad Sci U S A. 2010, 107(29), 13075-13080.
• References: [2] Herman SE, et al. Blood. 2011, 117(23), 6287-6296.
• References: [3] Chang BY, et al. Arthritis Res Ther. 2011, 13(4), R115.
• References: [4] Ponader S, et al. Blood. 2012, 119(5), 1182-1189.

REFERENCES

• Honigberg LA, et al. Proc Natl Acad Sci U S A. 2010, 107(29), 13075-13080.
• Herman SE, et al. Blood. 2011, 117(23), 6287-6296.
• Chang BY, et al. Arthritis Res Ther. 2011, 13(4), R115.
• Ponader S, et al. Blood. 2012, 119(5), 1182-1189.