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Ibrutinib (PCI-32765) Btk inhibitor

Name: Ibrutinib (PCI-32765)
Brand: Selleck Chemicals
SKU: S2680
Availability: InStock
Rating: 5 (543 reviews)

Cat.No.S2680

Ibrutinib is a potent and highly selective Brutons tyrosine kinase (Btk) inhibitor with IC50 of 0.5 nM in cell-free assays, modestly potent to Bmx, CSK, FGR, BRK, HCK, less potent to EGFR, Yes, ErbB2, JAK3, etc. Ibrutinib is applicable as a Btk ligand in the synthesis of a series of PROTACs including P13I.

Chemical Structure

Molecular Weight: 440.5

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Quality Control

Batch: Purity: 99.98%

99.98

Related Targets	EGFR VEGFR JAK PDGFR FGFR Src HIF FLT FLT3 HER2
Other Target Protein Ligand Inhibitors	(+)-JQ1 Quizartinib (AC220) BI-4464

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
human Rec1 cells	Function assay	2.5 μM	6 h	Inhibition of Lyn phosphorylation in human Rec1 cells at 2.5 uM incubated for 6 hrs by Western blotting method	25222877
human WSU-NHL cells	Cytotoxic assay		72 h	Cytotoxicity against human WSU-NHL cells assessed as growth inhibition after 72 hrs by CellTiter-Glo luminescent cell viability assay, IC50=1.09 μM	24915291
human SU-DHL6 cells	Cytotoxic assay		72 h	Cytotoxicity against human SU-DHL6 cells assessed as growth inhibition after 72 hrs by CellTiter-Glo luminescent cell viability assay, IC50=0.58 μM.	24915291
human DOHH2 cells	Cytotoxic assay		72 h	Cytotoxicity against human DOHH2 cells assessed as growth inhibition after 72 hrs by CellTiter-Glo luminescent cell viability assay, IC50=0.41 μM.	24915291
Sf9 cells	Function assay		1 h	Inhibition of LYN-A expressed in Sf9 cells after 60 mins by TR-FRET Assay, IC50=0.2 μM.	21958547
human Ramos cells	Function assay		1 h	Inhibition of Btk in human Ramos cells assessed as inhibition of PLC-gamma2 phosphorylation at Tyr1217 after 1 hr by Western blot analysis, IC50=14 nM.	24915291
human Pfeiffer cells	Function assay		72 h	Cytotoxicity against human Pfeiffer cells assessed as growth inhibition after 72 hrs by CellTiter-Glo luminescent cell viability assay, IC50=2 nM.	24915291
Sf9 cells	Function assay		1 h	Inhibition of human full-length BTK expressed in Sf9 cells using FAM-Srctide peptide as substrate after 60 mins by TR-FRET Assay， IC50=0.5 nM.	21958547
Sf9 insect cells	Function assay		60 mins	IC50 = 0.0003 μM	29146136
sf9 cells	Function assay			IC50 = 0.0003 μM	27994736
Sf9 insect cells	Function assay		60 mins	IC50 = 0.00034 μM	27912175
Sf9 insect cells	Function assay		60 mins	IC50 = 0.00034 μM	28432946
Sf9 insect cells	Function assay		60 mins	IC50 = 0.00034 μM	27956037
Sf9 cells	Function assay		60 mins	IC50 = 0.0004 μM	30006143
Sf9 cells	Function assay		60 mins	IC50 = 0.0005 μM	21958547
Ramos cells	Function assay		1 h	IC50 = 0.0005 μM	28280261
Sf9 cells	Function assay		60 mins	IC50 = 0.001 μM	30077608
Pfeiffer cells	Cytotoxicity assay		72 h	GI50 = 0.002 μM	24915291
B cells	Function assay		1 h	IC50 = 0.0046 μM	30290988
Sf9 insect cells	Function assay		2 to 60 mins	Ki = 0.0048 μM	28315597
Ramos cells	Function assay		1 h	IC50 = 0.0075 μM	24915291
Sf9 insect cells	Function assay		60 mins	IC50 = 0.008 μM	28315597
TMD8 cells	Antiproliferative activity assay		72 h	IC50 = 0.01 μM	29715023
CD19+ B cells	Function assay		1 h	IC50 = 0.012 μM	29457982
Sf9 insect cells	Function assay		60 mins	IC50 = 0.012 μM	28315597
Ramos cells	Function assay		1 h	IC50 = 0.014 μM	24915291
Sf9 insect cells	Function assay		1 h	IC50 = 0.0144 μM	29715023
Sf9 insect cells	Function assay		60 mins	IC50 = 0.0161 μM	29146136
HCC827 cells	Antiproliferative activity assay		72 h	IC50 = 0.039 μM	28734581
PC9 cells	Function assay		72 h	GI50 = 0.05 μM	28282122
Sf9 cells	Function assay		60 mins	IC50 = 0.1 μM	30006143
H3255 cells	Function assay		72 h	GI50 = 0.11 μM	28282122
BaF3 cells	Function assay		72 h	GI50 = 0.12 μM	26630553
BAF3 cells	Antiproliferative activity assay		72 h	GI50 = 0.12 μM	28956923
Sf9 insect cells	Function assay		60 mins	IC50 = 0.123 μM	28315597
Sf9 insect cells	Function assay		60 mins	IC50 = 0.146 μM	28315597
BAF3 cells	Function assay		72 h	GI50 = 0.16 μM	28282122
Sf9 cells	Function assay		60 mins	IC50 = 0.2 μM	21958547
MV411 cells	Growth inhibition assay			GI50 = 0.25 μM	28315597
MV4-11 cells	Antiproliferative activity assay		72 h	GI50 = 0.33 μM	26630553
MV4-11 cells	Antiproliferative activity assay		72 h	GI50 = 0.33 μM	28956923
DOHH2 cells	Cytotoxicity assay		72 h	GI50 = 0.41 μM	24915291
HCC827 cells	Antiproliferative activity assay		96 h	EC50 = 0.45 μM	28853575
SU-DHL6 cells	Cytotoxicity assay		72 h	GI50 = 0.58 μM	24915291
M07e cells	Growth inhibition assay			GI50 = 0.59 μM	28315597
NCI-H1975 cells	Antiproliferative activity assay		96 h	EC50 = 0.64 μM	28853575
SU-DHL-2 cells	Growth inhibition assay			GI50 = 0.64 μM	28315597
HEK293T cells	Function assay		1 h	IC50 = 0.9 μM	28280261
Ramos cells	Antiproliferative activity assay		72 h	IC50 = 0.92 μM	29715023
BA/F3 cells	Antiproliferative activity assay		72 h	IC50 = 1 μM	26258521
WSU-NHL cells	Cytotoxicity assay		72 h	GI50 = 1.09 μM	24915291
NCI-H1975 cells	Function assay		72 h	GI50 = 1.2 μM	28282122
NCI-H1975 cells	Antiproliferative activity assay		72 h	IC50 = 1.27 μM	28734581
Raji cells	Antiproliferative activity assay		48 h	IC50 = 1.49 μM	29567295
Pfeiffer cells	Growth inhibition assay			GI50 = 1.6 μM	28315597
A431 cells	Antiproliferative activity assay		96 h	EC50 = 2.38 μM	28853575
BAF3 cells	Antiproliferative activity assay		72 h	GI50 = 2.5 μM	28956923
U937 cells	Growth inhibition assay			GI50 = 2.9 μM	28315597
NB4 cells	Growth inhibition assay			GI50 = 3 μM	28315597
Ramos cells	Growth inhibition assay			GI50 = 3.4 μM	28315597
SKM1 cells	Growth inhibition assay			GI50 = 3.6 μM	28315597
U2932 cells	Growth inhibition assay			GI50 = 4.4 μM	28315597
Ramos cells	Antiproliferative activity assay		72 h	IC50 = 5.14 μM	30006143
Ramos cells	Antiproliferative activity assay		48 h	IC50 = 5.14 μM	27956037
Ramos cells	Antiproliferative activity assay		48 h	IC50 = 5.88 μM	28432946
Ramos cells	Antiproliferative activity assay		72 h	IC50 = 6.62 μM	29146136
K562 cells	Antiproliferative activity assay		48 h	IC50 = 7.5 μM	30077608
HL60 cells	Antiproliferative activity assay		48 h	IC50 = 8 μM	30077608
Ramos cells	Antiproliferative activity assay		48 h	IC50 = 8.11 μM	27994736
Ramos cells	Antiproliferative activity assay		48 h	IC50 = 8.26 μM	29567295
OCI-AML3 cells	Growth inhibition assay			GI50 = 9.2 μM	28315597
BAF3 cells	Cytotoxicity assay		72 h	GI50 = 10 μM	26630553
BAF3 cells	Antiproliferative activity assay		72 h	GI50 = 10 μM	28956923
BAF3 cells	Growth inhibition assay		72 h	GI50 = 10 μM	28282122
NAMALWA cells	Antiproliferative activity assay		72 h	IC50 = 10.45 μM	29146136
Ramos cells	Antiproliferative activity assay		48 h	IC50 = 12.6 μM	27912175
Raji cells	Antiproliferative activity assay		48 h	IC50 = 14.2 μM	30077608
Raji cells	Antiproliferative activity assay		48 h	IC50 = 15.2 μM	27994736
MIAPaCa2 cells	Cytotoxicity assay		3 days	IC50 = 16.6 μM	27077228
HeLa cells	Cytotoxicity assay		3 days	IC50 = 16.8 μM	27077228
Raji cells	Antiproliferative activity assay		48 h	IC50 = 19.3 μM	27912175
Raji cells	Antiproliferative activity assay		48 h	IC50 = 19.3 μM	28432946
Raji cells	Antiproliferative activity assay		72 h	IC50 = 19.5 μM	30006143
Raji cells	Antiproliferative activity assay		48 h	IC50 = 19.5 μM	27956037
NAMALWA cells	Antiproliferative activity assay		72 h	IC50 = 19.6 μM	30006143
A2780 cells	Cytotoxicity assay		3 days	EC50 = 20.1 μM	27077228
Raji cells	Antiproliferative activity assay		72 h	IC50 = 20.88 μM	29146136
A549 cells	Antiproliferative activity assay		72 h	IC50 = 21.79 μM	28734581
SW480 cells	Cytotoxicity assay		3 days	IC50 = 25.6 μM	27077228
Ramos cells	Cytotoxicity assay		24 h	IC50 = 28.7 μM	28274675
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 88 mg/mL (199.77 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 8 mg/mL

Water : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
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.
Clear solution	2%DMSO 1 40%PEG300 2 5%Tween80 3 53%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	1.760mg/ml (4.00mM)	Taking the 1 mL working solution as an example, add 20 μL 88 mg/ml clarified DMSO stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween80 to the above system, mix evenly to clarify it; then continue to add 530 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	7.500mg/ml (17.03mM)	Taking the 1 mL working solution as an example, add 50 μL of 150 mg/ml clarified DMSO stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	1.650mg/ml (3.75mM)	Taking the 1 mL working solution as an example, add 50 μL of 33 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
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.
Clear solution	5%DMSO 1 30%PEG300 2 5%Tween80 3 60%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	10.000mg/ml (22.70mM)	Taking the 1 mL working solution as an example, add 50 μL of 200 mg/ml clarified DMSO stock solution to 300 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 600 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
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.
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.
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.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg

Average weight of animals: g

Dosing volume per animal: μL

Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO

% Tween 80

% ddH₂O

% DMSO

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Chemical Information, Storage & Stability

Molecular Weight	440.5	Formula	C₂₅H₂₄N₆O₂	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	936563-96-1	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	PCI-32765			Smiles	C=CC(=O)N1CCCC(C1)N2C3=NC=NC(=C3C(=N2)C4=CC=C(C=C4)OC5=CC=CC=C5)N

Mechanism of Action

Targets/IC₅₀/Ki	BTK (Cell-free assay) 0.5 nM BLK (Cell-free assay) 0.5 nM Bmx (Cell-free assay) 0.8 nM CSK (Cell-free assay) 2.3 nM FGR (Cell-free assay) 2.3 nM BRK (Cell-free assay) 3.3 nM Hck (Cell-free assay) 3.7 nM EGFR (Cell-free assay) 5.6 nM YES (Cell-free assay) 6.5 nM ErbB2 (Cell-free assay) 9.4 nM ITK (Cell-free assay) 10.7 nM JAK3 (Cell-free assay) 16.1 nM FRK (Cell-free assay) 29.2 nM LCK (Cell-free assay) 33.2 nM RET (Cell-free assay) 36.5 nM
In vitro	Ibrutinib shows the potent and irreversible inhibitory effect and selectivity for Btk enzymatic activity. In BCR pathway-activated DOHH2 cell line, this compound 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. It exhibits a significant dose-dependent and time-dependent induction of cytotoxicity in chronic lymphocytic leukemia (CLL) cells. In addition, this compound induces cell death depending on caspase pathway activation and antagonizes the ability of CLL cells to proliferate after TLR signaling. A recent study shows that this chemical 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.
Kinase Assay	Kinase Assays
Kinase Assay	In vitro kinase IC50 values are measured using ³³P filtration binding assay after 1 hour incubation of kinase, ³³P-ATP, Ibrutinib, and substrate [0.2 mg/mL poly(EY)(4:1]. Assays are performed at Reaction Biology.
In vivo	In a collagen-induced arthritis model, Ibrutinib significantly reduces clinical arthritis scores reflecting paw swelling and joint inflammation by inhibiting B-cell activation. In a MRL-Fas(lpr) lupus model, this compound reduces renal disease and autoantibody production. In an adoptive transfer TCL1 mouse model of CLL, this compound (25 mg/kg/day) causes a transient early lymphocytosis, and delays CLL disease progression.
References	[1] https://pubmed.ncbi.nlm.nih.gov/20615965/ [2] https://pubmed.ncbi.nlm.nih.gov/21422473/ [3] https://pubmed.ncbi.nlm.nih.gov/21752263/ [4] https://pubmed.ncbi.nlm.nih.gov/22180443/

Applications

Methods	Biomarkers	PMID
Western blot	pEGFR(Tyr1068) / EGFR pBTK / pPLCγ2 / pAKT / pERK / pJNK	28061447
Immunofluorescence	CD11b COX-2	30231870
ELISA	hTNFα IL-10	26627823

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT06084923	Not yet recruiting	Chronic Lymphocytic Leukemia	Gruppo Italiano Malattie EMatologiche dell''Adulto	May 2024	--
NCT06224452	Not yet recruiting	Hematological Malignancy\|Atrial Fibrillation	University Hospital Caen	March 1 2024	--
NCT05694312	Recruiting	Autoimmune Hemolytic Anemia\|Chronic Lymphocytic Leukemia\|Small Lymphocytic Lymphoma\|Monoclonal B-Cell Lymphocytosis CLL-Type	Gruppo Italiano Malattie EMatologiche dell''Adulto	November 24 2023	Phase 2

Tech Support

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

Frequently Asked Questions

Question 1:
How to reconstitute it for in vivo studies?

Answer:
For in vivo study, we suggest to use 5% DMSO+30% PEG 300+5% Tween 80+ddH2O up to 10mg/ml for it.

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