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

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...

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Main Authors: ES Kolotova, VV Tatarskii, AA Zeifman, OV Stroganov, VS Stroilov, IYu Titov, FN Novikov, AA Kalinina, GG Chilov, AA Shtil’
Format: Article
Language:Russian
Published: Practical Medicine Publishing House 2016-01-01
Series:Клиническая онкогематология
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Online Access:http://bloodjournal.ru/en/pf-114-a-novel-inhibitor-of-bcr-abl-chimeric-tyrosine-kinase-attenuates-intracellular-crkl-phosphorylation-and-kills-chronic-myeloid-leukemia-cells/
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author ES Kolotova
VV Tatarskii
AA Zeifman
OV Stroganov
VS Stroilov
IYu Titov
FN Novikov
AA Kalinina
GG Chilov
AA Shtil’
author_facet ES Kolotova
VV Tatarskii
AA Zeifman
OV Stroganov
VS Stroilov
IYu Titov
FN Novikov
AA Kalinina
GG Chilov
AA Shtil’
author_sort ES Kolotova
collection DOAJ
description 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.
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spelling doaj-art-97b83956983546b4abd133eb6e2d4a9b2025-08-20T01:58:35ZrusPractical Medicine Publishing HouseКлиническая онкогематология1997-69332500-21392016-01-01911510.21320/2500-2139-2016-9-1-1-5PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia CellsES Kolotova0VV Tatarskii1AA Zeifman2OV Stroganov3VS Stroilov4IYu Titov5FN Novikov6AA Kalinina7GG Chilov8AA Shtil’9N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991; Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991; Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991; Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991;Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991; Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478N.D. Zelinskii Institute of Organic Chemistry, 47 Leninskii pr-t, Moscow, Russian Federation, 119991; Fusion Pharma, 18 bld. 2 Generala Dorokhova str., Moscow, Russian Federation, 119530N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478Background & 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.http://bloodjournal.ru/en/pf-114-a-novel-inhibitor-of-bcr-abl-chimeric-tyrosine-kinase-attenuates-intracellular-crkl-phosphorylation-and-kills-chronic-myeloid-leukemia-cells/chronic myeloid leukemiaBcr-Abl tyrosine kinaseprotein phosphorylationflow cytometrycytotoxicity
spellingShingle ES Kolotova
VV Tatarskii
AA Zeifman
OV Stroganov
VS Stroilov
IYu Titov
FN Novikov
AA Kalinina
GG Chilov
AA Shtil’
PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells
Клиническая онкогематология
chronic myeloid leukemia
Bcr-Abl tyrosine kinase
protein phosphorylation
flow cytometry
cytotoxicity
title PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells
title_full PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells
title_fullStr PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells
title_full_unstemmed PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells
title_short PF-114, a Novel Inhibitor of Bcr-Abl Chimeric Tyrosine Kinase, Attenuates Intracellular CrkL Phosphorylation and Kills Chronic Myeloid Leukemia Cells
title_sort pf 114 a novel inhibitor of bcr abl chimeric tyrosine kinase attenuates intracellular crkl phosphorylation and kills chronic myeloid leukemia cells
topic chronic myeloid leukemia
Bcr-Abl tyrosine kinase
protein phosphorylation
flow cytometry
cytotoxicity
url http://bloodjournal.ru/en/pf-114-a-novel-inhibitor-of-bcr-abl-chimeric-tyrosine-kinase-attenuates-intracellular-crkl-phosphorylation-and-kills-chronic-myeloid-leukemia-cells/
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