Biochemical pharmacology and resistance to 2-chloro-2'-arabinofluoro-2'- deoxyadenosine, a novel analogue of cladribine in human leukemic cells

Kourosh Lotfi, Emma Månsson, T. Spasokoukotskaja, Birgitta Pettersson, Jan Liliemark, Curt Peterson, Staffan Eriksson, Freidoun Albertioni

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

The objective of the present study was to investigate the biochemical pharmacology of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (CAFdA) - a fluorinated analogue of cladribine [2-chloro-2'-deoxyadenosine, Leustatin (CdA)] with improved acid and metabolic stability - in human leukemic cell lines and in mononuclear cells isolated from patients with chronic lymphocytic leukemia (CLL) and acute myelocytic leukemia (AML). We have also made and characterized two cell lines that are not sensitive to the growth inhibitory and cytotoxic effects of CAFdA. Incubation of cells isolated from the blood of CLL and AML patients with various concentrations of CdA or of CAFdA accumulated CdA and CAFdA nucleotides in a dose-dependent manner. A significantly higher rate of phosphorylation to monophosphates was observed for CAFdA than for CdA in cells from CLL patients (n = 14; P = 0.04). The differences in the phosphorylation were even more pronounced for the respective triphosphates in both CLL (n = 14; P = 0.001) and AML (n - 4; P = 0.04) cells. Retention of CAFdA 5'-triphosphate (CAFdATP) was also longer than that for CdA 5'-triphosphate (CdATP) in cells from leukemic patients. The relative efficacy of CAFdA as a substrate for purified recombinant deoxycytidine kinase (dCK), the key enzyme in the activation of nucleoside analogues, was very high and exceeded that of CdA as well as the natural substrate, deoxycytidine, by a factor of 2 and 8, respectively. The K(m) for CAFdA with dCK was also lower than that for CdA, as measured in crude extracts from the human acute lymphoblastic leukemia cell line CCRF-CEM and the promyelocytic leukemia cell line HL60. Acquired resistance to CAFdA in HL60 and in CCRF-CEM cell lines was directly correlated to the decreased activity of the nucleoside phosphorylating enzyme, dCK. Resistant cells also showed a considerable degree of cross-resistance to analogues that were activated by dCK. These observations demonstrated that dCK phosphorylates CAFdA more efficiently than CdA. Furthermore, CAFdATP is apparently more stable than CdATP and the mechanisms of resistance to CAFdA are similar to those leading to CdA resistance. These results encourage studies on the clinical effect of CAFdA in lymphoproliferative diseases.

Original languageEnglish
Pages (from-to)2438-2444
Number of pages7
JournalClinical Cancer Research
Volume5
Issue number9
Publication statusPublished - Sep 1999

Fingerprint

Cladribine
Pharmacology
Deoxycytidine Kinase
B-Cell Chronic Lymphocytic Leukemia
Cell Line
Acute Myeloid Leukemia
2'-chloro-2'-deoxyadenosine
Nucleosides
clofarabine
Phosphorylation

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Biochemical pharmacology and resistance to 2-chloro-2'-arabinofluoro-2'- deoxyadenosine, a novel analogue of cladribine in human leukemic cells. / Lotfi, Kourosh; Månsson, Emma; Spasokoukotskaja, T.; Pettersson, Birgitta; Liliemark, Jan; Peterson, Curt; Eriksson, Staffan; Albertioni, Freidoun.

In: Clinical Cancer Research, Vol. 5, No. 9, 09.1999, p. 2438-2444.

Research output: Contribution to journalArticle

Lotfi, K, Månsson, E, Spasokoukotskaja, T, Pettersson, B, Liliemark, J, Peterson, C, Eriksson, S & Albertioni, F 1999, 'Biochemical pharmacology and resistance to 2-chloro-2'-arabinofluoro-2'- deoxyadenosine, a novel analogue of cladribine in human leukemic cells', Clinical Cancer Research, vol. 5, no. 9, pp. 2438-2444.
Lotfi, Kourosh ; Månsson, Emma ; Spasokoukotskaja, T. ; Pettersson, Birgitta ; Liliemark, Jan ; Peterson, Curt ; Eriksson, Staffan ; Albertioni, Freidoun. / Biochemical pharmacology and resistance to 2-chloro-2'-arabinofluoro-2'- deoxyadenosine, a novel analogue of cladribine in human leukemic cells. In: Clinical Cancer Research. 1999 ; Vol. 5, No. 9. pp. 2438-2444.
@article{e0141649cc5a4f96a94f3af8fe1ef25a,
title = "Biochemical pharmacology and resistance to 2-chloro-2'-arabinofluoro-2'- deoxyadenosine, a novel analogue of cladribine in human leukemic cells",
abstract = "The objective of the present study was to investigate the biochemical pharmacology of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (CAFdA) - a fluorinated analogue of cladribine [2-chloro-2'-deoxyadenosine, Leustatin (CdA)] with improved acid and metabolic stability - in human leukemic cell lines and in mononuclear cells isolated from patients with chronic lymphocytic leukemia (CLL) and acute myelocytic leukemia (AML). We have also made and characterized two cell lines that are not sensitive to the growth inhibitory and cytotoxic effects of CAFdA. Incubation of cells isolated from the blood of CLL and AML patients with various concentrations of CdA or of CAFdA accumulated CdA and CAFdA nucleotides in a dose-dependent manner. A significantly higher rate of phosphorylation to monophosphates was observed for CAFdA than for CdA in cells from CLL patients (n = 14; P = 0.04). The differences in the phosphorylation were even more pronounced for the respective triphosphates in both CLL (n = 14; P = 0.001) and AML (n - 4; P = 0.04) cells. Retention of CAFdA 5'-triphosphate (CAFdATP) was also longer than that for CdA 5'-triphosphate (CdATP) in cells from leukemic patients. The relative efficacy of CAFdA as a substrate for purified recombinant deoxycytidine kinase (dCK), the key enzyme in the activation of nucleoside analogues, was very high and exceeded that of CdA as well as the natural substrate, deoxycytidine, by a factor of 2 and 8, respectively. The K(m) for CAFdA with dCK was also lower than that for CdA, as measured in crude extracts from the human acute lymphoblastic leukemia cell line CCRF-CEM and the promyelocytic leukemia cell line HL60. Acquired resistance to CAFdA in HL60 and in CCRF-CEM cell lines was directly correlated to the decreased activity of the nucleoside phosphorylating enzyme, dCK. Resistant cells also showed a considerable degree of cross-resistance to analogues that were activated by dCK. These observations demonstrated that dCK phosphorylates CAFdA more efficiently than CdA. Furthermore, CAFdATP is apparently more stable than CdATP and the mechanisms of resistance to CAFdA are similar to those leading to CdA resistance. These results encourage studies on the clinical effect of CAFdA in lymphoproliferative diseases.",
author = "Kourosh Lotfi and Emma M{\aa}nsson and T. Spasokoukotskaja and Birgitta Pettersson and Jan Liliemark and Curt Peterson and Staffan Eriksson and Freidoun Albertioni",
year = "1999",
month = "9",
language = "English",
volume = "5",
pages = "2438--2444",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "9",

}

TY - JOUR

T1 - Biochemical pharmacology and resistance to 2-chloro-2'-arabinofluoro-2'- deoxyadenosine, a novel analogue of cladribine in human leukemic cells

AU - Lotfi, Kourosh

AU - Månsson, Emma

AU - Spasokoukotskaja, T.

AU - Pettersson, Birgitta

AU - Liliemark, Jan

AU - Peterson, Curt

AU - Eriksson, Staffan

AU - Albertioni, Freidoun

PY - 1999/9

Y1 - 1999/9

N2 - The objective of the present study was to investigate the biochemical pharmacology of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (CAFdA) - a fluorinated analogue of cladribine [2-chloro-2'-deoxyadenosine, Leustatin (CdA)] with improved acid and metabolic stability - in human leukemic cell lines and in mononuclear cells isolated from patients with chronic lymphocytic leukemia (CLL) and acute myelocytic leukemia (AML). We have also made and characterized two cell lines that are not sensitive to the growth inhibitory and cytotoxic effects of CAFdA. Incubation of cells isolated from the blood of CLL and AML patients with various concentrations of CdA or of CAFdA accumulated CdA and CAFdA nucleotides in a dose-dependent manner. A significantly higher rate of phosphorylation to monophosphates was observed for CAFdA than for CdA in cells from CLL patients (n = 14; P = 0.04). The differences in the phosphorylation were even more pronounced for the respective triphosphates in both CLL (n = 14; P = 0.001) and AML (n - 4; P = 0.04) cells. Retention of CAFdA 5'-triphosphate (CAFdATP) was also longer than that for CdA 5'-triphosphate (CdATP) in cells from leukemic patients. The relative efficacy of CAFdA as a substrate for purified recombinant deoxycytidine kinase (dCK), the key enzyme in the activation of nucleoside analogues, was very high and exceeded that of CdA as well as the natural substrate, deoxycytidine, by a factor of 2 and 8, respectively. The K(m) for CAFdA with dCK was also lower than that for CdA, as measured in crude extracts from the human acute lymphoblastic leukemia cell line CCRF-CEM and the promyelocytic leukemia cell line HL60. Acquired resistance to CAFdA in HL60 and in CCRF-CEM cell lines was directly correlated to the decreased activity of the nucleoside phosphorylating enzyme, dCK. Resistant cells also showed a considerable degree of cross-resistance to analogues that were activated by dCK. These observations demonstrated that dCK phosphorylates CAFdA more efficiently than CdA. Furthermore, CAFdATP is apparently more stable than CdATP and the mechanisms of resistance to CAFdA are similar to those leading to CdA resistance. These results encourage studies on the clinical effect of CAFdA in lymphoproliferative diseases.

AB - The objective of the present study was to investigate the biochemical pharmacology of 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (CAFdA) - a fluorinated analogue of cladribine [2-chloro-2'-deoxyadenosine, Leustatin (CdA)] with improved acid and metabolic stability - in human leukemic cell lines and in mononuclear cells isolated from patients with chronic lymphocytic leukemia (CLL) and acute myelocytic leukemia (AML). We have also made and characterized two cell lines that are not sensitive to the growth inhibitory and cytotoxic effects of CAFdA. Incubation of cells isolated from the blood of CLL and AML patients with various concentrations of CdA or of CAFdA accumulated CdA and CAFdA nucleotides in a dose-dependent manner. A significantly higher rate of phosphorylation to monophosphates was observed for CAFdA than for CdA in cells from CLL patients (n = 14; P = 0.04). The differences in the phosphorylation were even more pronounced for the respective triphosphates in both CLL (n = 14; P = 0.001) and AML (n - 4; P = 0.04) cells. Retention of CAFdA 5'-triphosphate (CAFdATP) was also longer than that for CdA 5'-triphosphate (CdATP) in cells from leukemic patients. The relative efficacy of CAFdA as a substrate for purified recombinant deoxycytidine kinase (dCK), the key enzyme in the activation of nucleoside analogues, was very high and exceeded that of CdA as well as the natural substrate, deoxycytidine, by a factor of 2 and 8, respectively. The K(m) for CAFdA with dCK was also lower than that for CdA, as measured in crude extracts from the human acute lymphoblastic leukemia cell line CCRF-CEM and the promyelocytic leukemia cell line HL60. Acquired resistance to CAFdA in HL60 and in CCRF-CEM cell lines was directly correlated to the decreased activity of the nucleoside phosphorylating enzyme, dCK. Resistant cells also showed a considerable degree of cross-resistance to analogues that were activated by dCK. These observations demonstrated that dCK phosphorylates CAFdA more efficiently than CdA. Furthermore, CAFdATP is apparently more stable than CdATP and the mechanisms of resistance to CAFdA are similar to those leading to CdA resistance. These results encourage studies on the clinical effect of CAFdA in lymphoproliferative diseases.

UR - http://www.scopus.com/inward/record.url?scp=0345466497&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0345466497&partnerID=8YFLogxK

M3 - Article

C2 - 10499616

AN - SCOPUS:0345466497

VL - 5

SP - 2438

EP - 2444

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 9

ER -