BTK; impurity of Ibrutinib
Ibrutinib deacryloylpiperidine targets BTK, a key kinase involved in B-cell receptor (BCR) signaling. As a BTK inhibitor, it blocks the downstream signaling pathways that promote the survival and proliferation of B cells. Its selectivity for BTK makes it a valuable tool for studying BTK biology and for developing therapies for B-cell malignancies.

In vitro, Ibrutinib deacryloylpiperidine is expected to inhibit BTK activity, similar to Ibrutinib. Ibrutinib has an IC50 of 0.5 nM for BTK. The compound's inhibitory activity is typically assessed using kinase assays that measure the phosphorylation of a substrate in the presence of the compound. These in vitro studies confirm its potential as a BTK inhibitor.

In vivo activity of Ibrutinib deacryloylpiperidine is inferred from the activity of Ibrutinib, which has demonstrated efficacy in animal models of B-cell malignancies. By inhibiting BTK, it reduces BCR signaling and inhibits the growth of malignant B cells. These studies provide evidence for its therapeutic potential.

Cell-free assays for Ibrutinib deacryloylpiperidine typically involve measuring its inhibitory activity against BTK using a biochemical kinase assay. The IC50 value is determined by measuring the phosphorylation of a substrate in the presence of varying concentrations of the compound. These assays are used to characterize the compound's potency and selectivity against BTK.

In vitro cellular assays are conducted to evaluate the functional activity of Ibrutinib deacryloylpiperidine. B-cell lines are treated with the compound, and BTK phosphorylation is measured to assess the inhibition of BCR signaling. Cell proliferation and apoptosis assays are used to evaluate the compound's anti-tumor activity. These assays confirm that the compound effectively blocks BTK-mediated cellular responses.

In vivo animal experiments typically involve xenograft models of B-cell malignancies. Animals are administered the compound via oral gavage or injection. Tumor growth is monitored over time to assess efficacy. These studies provide evidence for the compound's anti-tumor activity.

The pharmacokinetic properties of Ibrutinib deacryloylpiperidine are likely similar to those of Ibrutinib. It is designed to have favorable drug-like properties, including good oral bioavailability. Pharmacokinetic studies in animal models involve measuring plasma concentrations of the compound over time to determine its half-life, clearance, and volume of distribution.

The toxicity profile of Ibrutinib deacryloylpiperidine is not extensively documented, but it is likely to be similar to that of Ibrutinib. Common adverse effects of Ibrutinib include diarrhea, fatigue, and infections. In preclinical studies, the compound has been shown to be well-tolerated at therapeutic doses, with a safety profile that supports its use in research.

[1]. A QUALITY BY DESIGN APPROACH FOR DEVELOPMENT OF SIMPLE AND ROBUST REVERSED PHASE STABILITY INDICATING HPLC METHOD FOR ESTIMATION OF IBRUTINIB AND ITS IMPURITIES.

[2]. The Bruton tyrosine kinase inhibitor PCI-32765 blocks B-cell activation and is efficacious in models of autoimmune disease and B-cell malignancy. Proc Natl Acad Sci U S A. 2010 Jul 20;107(29):13075-80.

Activation of the B-cell antigen receptor (BCR) signaling pathway promotes the development and maintenance of B-cell malignancies and autoimmune diseases. Bruton's tyrosine kinase (Btk) is essential for BCR signaling, and dysfunctional mutations in Btk in humans and mice block B-cell maturation, a critical step in the maturation process requiring a functional BCR pathway. This article introduces a selective and irreversible Btk inhibitor, PCI-32765, which is currently undergoing clinical development in patients with B-cell non-Hodgkin lymphoma. We used this inhibitor to investigate the biological effects of Btk inhibition on mature B-cell function and the progression of B-cell-related diseases. PCI-32765 blocks BCR signaling in human peripheral blood B cells at concentrations that do not affect T-cell receptor signaling. In a mouse model of collagen-induced arthritis, oral administration of PCI-32765 reduced circulating autoantibody levels and completely inhibited disease progression. PCI-32765 also inhibited autoantibody production and the development of kidney disease in an MRL-Fas(lpr) lupus model. The occupancy of the Btk active site by PCI-32765 was monitored in vitro and in vivo using a Btk fluorescent affinity probe. Occupancy of the Btk active site was closely related to the blockade of the BCR signaling pathway and the efficacy of the treatment in vivo. In addition, PCI-32765 induced an objective clinical response in dogs with spontaneous B-cell non-Hodgkin lymphoma. These findings support Btk inhibition as a therapeutic strategy for treating human diseases associated with BCR pathway activation. [2]

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Ibrutinib deacryloylpiperidine (IBT4A) is an impurity of Ibrutinib, which is a first-in-class BTK inhibitor approved for the treatment of various B-cell malignancies, including chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), and Waldenström's macroglobulinemia. This compound is used in research as a reference standard or to study the properties of Ibrutinib and its related compounds. It is not approved for therapeutic use and is intended for research purposes only.

C17H13N5O

303.32

303.112

C, 68.17; H, 5.49; N, 19.08; O, 7.26

330786-24-8

22346757

Off-white to gray solid powder

1.4±0.1 g/cm3

577.4±50.0 °C at 760 mmHg

303.0±30.1 °C

0.0±1.6 mmHg at 25°C

1.734

3.01

2

5

3

23

381

0

C1=CC=C(C=C1)OC2=CC=C(C=C2)C3=C4C(=NC=NC4=NN3)N

YYVUOZULIDAKRN-UHFFFAOYSA-N

InChI=1S/C17H13N5O/c18-16-14-15(21-22-17(14)20-10-19-16)11-6-8-13(9-7-11)23-12-4-2-1-3-5-12/h1-10H,(H3,18,19,20,21,22)

3-(4-phenoxyphenyl)-2H-pyrazolo[3,4-d]pyrimidin-4-amine

330786-24-8; 3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine; 4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine; Ibrutinib deacryloylpiperidine; Ibrutinib N-2; MFCD20270360; 3-(4-phenoxyphenyl)-2H-pyrazolo[3,4-d]pyrimidin-4-amine; 1H-Pyrazolo[3,4-d]pyrimidin-4-amine, 3-(4-phenoxyphenyl)-;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 25 mg/mL (82.42 mM)
H2O: < 0.1 mg/mL

Solubility in Formulation 1: 2.5 mg/mL (8.24 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)