Characterization of compound 584, an Abl kinase inhibitor with lasting effects

Miriam Puttini, Sara Redaelli, Loris Moretti, Stefania Brussolo, Rosalind H. Gunby, Luca Mologni, Edoardo Marchesi, Loredana Cleris, Arianna Donella-Deana, Peter Drueckes, Elisa Sala, Vittorio Lucchini, Michael Kubbutat, Franca Formelli, Alfonso Zambon, Leonardo Scapozza, Carlo Gambacorti-Passerini

Research output: Contribution to journalArticle

Abstract

Background: Resistance to imatinib is an important clinical issue in the treatment of Philadelphia chromosomepositive leukemias which is being tackled by the development of new, more potent drugs, such as the dual Src/Abl tyrosine kinase inhibitors dasatinib and bosutinib and the imatinib analog nilotinib. In the current study we describe the design, synthesis and biological properties of an imatinib analog with a chlorine-substituted benzamide, namely compound 584 (cmp-584). Design and Methods: To increase the potency, we rationally designed cmp-584, a compound with enhanced shape complementarity with the kinase domain of Abl. cmp-584 was synthesized and characterized in vitro against a panel of 67 serine/threonine and tyrosine kinases using radioactive and enzyme-linked immunosorbent kinase assays. We studied inhibitory cellular activity using Bcr/Abl-positive human cell lines, murine transfectants in proliferation experiments, and a murine xenotransplanted model. Kinase assays on isolated Bcr/Abl protein were also performed. Finally, we used a wash-out approach on whole cells to study the binding kinetics of the inhibitor. Results: cmp-584 showed potent anti-Abl activity both on recombinant protein (IC50: 8 nM) and in cell-based assays (IC50: 0.1-10 nM). The drug maintained inhibitory activity against platelet-derived growth factor receptors and c-KIT and was also active against Lyn (IC50: 301 nM). No other kinase of the panel was inhibited at nanomolar doses. cmp-584 was 20- to 300-fold more active than imatinib in cells. This superior activity was evident in intact cells, in which full-length Bcr-Abl is present. In vivo experiments confirmed the activity of cmp-584. Wash-out experiments showed that short exposure to the drug impaired cell proliferation and Bcr-Abl phosphorylation for a substantially longer period of time than imatinib. Conclusions: The present results suggest a slower off-rate (dissociation rate) of cmp-584 compared to imatinib as an explanation for the increased cellular activity of the former.

Original languageEnglish
Pages (from-to)653-661
Number of pages9
JournalHaematologica
Volume93
Issue number5
DOIs
Publication statusPublished - May 2008

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Phosphotransferases
Inhibitory Concentration 50
Pharmaceutical Preparations
Platelet-Derived Growth Factor Receptors
src-Family Kinases
Protein-Serine-Threonine Kinases
Chlorine
Recombinant Proteins
Protein-Tyrosine Kinases
Imatinib Mesylate
Leukemia
Enzyme-Linked Immunosorbent Assay
Phosphorylation
Cell Proliferation
Cell Line
Proteins
Therapeutics

Keywords

  • Abl
  • Imatinib analog
  • Off-rate
  • Tyrosine kinase inhibitor

ASJC Scopus subject areas

  • Hematology

Cite this

Puttini, M., Redaelli, S., Moretti, L., Brussolo, S., Gunby, R. H., Mologni, L., ... Gambacorti-Passerini, C. (2008). Characterization of compound 584, an Abl kinase inhibitor with lasting effects. Haematologica, 93(5), 653-661. https://doi.org/10.3324/haematol.12212

Characterization of compound 584, an Abl kinase inhibitor with lasting effects. / Puttini, Miriam; Redaelli, Sara; Moretti, Loris; Brussolo, Stefania; Gunby, Rosalind H.; Mologni, Luca; Marchesi, Edoardo; Cleris, Loredana; Donella-Deana, Arianna; Drueckes, Peter; Sala, Elisa; Lucchini, Vittorio; Kubbutat, Michael; Formelli, Franca; Zambon, Alfonso; Scapozza, Leonardo; Gambacorti-Passerini, Carlo.

In: Haematologica, Vol. 93, No. 5, 05.2008, p. 653-661.

Research output: Contribution to journalArticle

Puttini, M, Redaelli, S, Moretti, L, Brussolo, S, Gunby, RH, Mologni, L, Marchesi, E, Cleris, L, Donella-Deana, A, Drueckes, P, Sala, E, Lucchini, V, Kubbutat, M, Formelli, F, Zambon, A, Scapozza, L & Gambacorti-Passerini, C 2008, 'Characterization of compound 584, an Abl kinase inhibitor with lasting effects', Haematologica, vol. 93, no. 5, pp. 653-661. https://doi.org/10.3324/haematol.12212
Puttini M, Redaelli S, Moretti L, Brussolo S, Gunby RH, Mologni L et al. Characterization of compound 584, an Abl kinase inhibitor with lasting effects. Haematologica. 2008 May;93(5):653-661. https://doi.org/10.3324/haematol.12212
Puttini, Miriam ; Redaelli, Sara ; Moretti, Loris ; Brussolo, Stefania ; Gunby, Rosalind H. ; Mologni, Luca ; Marchesi, Edoardo ; Cleris, Loredana ; Donella-Deana, Arianna ; Drueckes, Peter ; Sala, Elisa ; Lucchini, Vittorio ; Kubbutat, Michael ; Formelli, Franca ; Zambon, Alfonso ; Scapozza, Leonardo ; Gambacorti-Passerini, Carlo. / Characterization of compound 584, an Abl kinase inhibitor with lasting effects. In: Haematologica. 2008 ; Vol. 93, No. 5. pp. 653-661.
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AU - Mologni, Luca

AU - Marchesi, Edoardo

AU - Cleris, Loredana

AU - Donella-Deana, Arianna

AU - Drueckes, Peter

AU - Sala, Elisa

AU - Lucchini, Vittorio

AU - Kubbutat, Michael

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