PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells

EXPERIMENTAL STUDIES

ES Kolotova1, VV Tatarskii1, AA Zeifman2,3, OV Stroganov2,3, VS Stroilov2,3, IYu Titov2,3, FN Novikov2,3, AA Kalinina1, GG Chilov2,3, AA Shtil’1

1 N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478

2 N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991

3 Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530

For correspondence: Aleksandr Al’bertovich Shtil’, DSci, 24 Kashirskoye sh., Moscow, Russian Federation, 115478; Tel.: + 7(499)612-78-34; e-mail: shtilaa@yahoo.com

For citation: Kolotova ES, Tatarskii VV, Zeifman AA, et al. PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells. Clinical oncohematology. 2016;9(1):1–5 (In Russ).

DOI: 10.21320/2500-2139-2016-9-1-1-5

ABSTRACT

Background & Aims. The chimeric tyrosine kinase Bcr-Abl triggers malignant transformation of myeloid cells via phosphorylation of a number of substrates including the CrkL adaptor protein. Pharmacological inhibition of Bcr-Abl mediated signaling is a major strategy in treatment of patients with chronic myeloid leukemia (CML). A new specific Bcr-Abl inhibitor (PF-114) was designed using a molecular modeling approach. The paper defines the cytotoxicity of PF-114 against CML cells and its effect on the CrkL phosphorylation.

Methods. The cytotoxicity was determined using the MTT assay. The total intracellular CrKL pool (phosphorylated and non-phosphorylated forms) was determined by means of flow cytometry.

Results. Exposure of Bcr-Abl-positive, K562 cell line to PF-114 blocked intracellular CrkL phosphorylation and caused cell death. In contrast, virtually no phosphorylated CrkL was detectable in Bcr-Abl-negative HL60, U937 and Jurkat leukemia cell lines.

Conclusion. Absence of phosphorylation in Bcr-Abl-negative cells (HL60, U937 and Jurkat) and death of HL60 cells under the effect of PF-114 at concentrations exceeding those required to kill K562 cells supports the emergence of PF-114 as a promising drug candidate for CML.

Keywords: chronic myeloid leukemia, Bcr-Abl tyrosine kinase, protein phosphorylation, flow cytometry, cytotoxicity.

Received: September 15, 2015

Accepted: October 8, 2015

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