TY - JOUR
T1 - Cellular pharmacokinetics and pharmacodynamics of the deoxycytidine analog 2′-C-cyano-2′-deoxy-1-β-D-arabino-pentofuranosylcytosine (CNDAC)
AU - Azuma, Atsushi
AU - Huang, Peng
AU - Matsuda, Akira
AU - Plunkett, Williamm
N1 - Funding Information:
This work was supported, in part, by Grant CA28596 from the National Cancer Institute, Department of Health and Human Safety and Cancer Center Support Grant P30 CA16672, and by Sankyo Co. Ltd., Tokyo, Japan.
PY - 2001/6/15
Y1 - 2001/6/15
N2 - The pharmacokinetics and pharmacodynamics of the novel clinical candidate 2′-C-cyano-2′-deoxy-1-β-D-arabino-pentofuranosylcytosine (CNDAC) were investigated in human lymphoblastoid CCRF-CEM cells and human myeloblastic leukemia ML-1 cells. Formation of CNDAC 5′-mono-, di-, and triphosphate (CNDACTP) was concentration-dependent; nucleotide accumulation was greater in the lymphoid cells than in the myeloid cells. The nucleotides were eliminated with linear kinetics from both lines, but were retained more effectively by the ML-1 cells. DNA synthesis was selectively inhibited by a 4-hr treatment with CNDAC in CCRF-CEM and ML-1 cells; the IC50 values were 1 and 0.8 μM, respectively. Evaluation of the polymerization reaction of a primer on an M13mp19(+) template by human DNA polymerase α indicated that CNDACTP was incorporated effectively (Km = 0.22 μM) opposite a complementary dGMP in the template strand. CNDACTP competed with the normal substrate, dCTP, for incorporation, and the two nucleotides showed similar substrate efficiencies (Vmax/Km: dCTP = 0.91; CNDACTP = 0.77). Primer extension was potently inhibited by CNDAC triphosphate (Ki = 23 nM); once the analog had been incorporated, further extension was not observed in vitro, suggesting that primers containing a 3′-terminal nucleotide analog were high Km substrates for polymerase α. Thus, the ability of human leukemia cells to effectively accumulate and retain CNDACTP, coupled with the favorable kinetics of competition for incorporation into DNA, and the relatively strong ability of the analog to terminate further extension, are likely to contribute to the cytotoxic action of CNDAC.
AB - The pharmacokinetics and pharmacodynamics of the novel clinical candidate 2′-C-cyano-2′-deoxy-1-β-D-arabino-pentofuranosylcytosine (CNDAC) were investigated in human lymphoblastoid CCRF-CEM cells and human myeloblastic leukemia ML-1 cells. Formation of CNDAC 5′-mono-, di-, and triphosphate (CNDACTP) was concentration-dependent; nucleotide accumulation was greater in the lymphoid cells than in the myeloid cells. The nucleotides were eliminated with linear kinetics from both lines, but were retained more effectively by the ML-1 cells. DNA synthesis was selectively inhibited by a 4-hr treatment with CNDAC in CCRF-CEM and ML-1 cells; the IC50 values were 1 and 0.8 μM, respectively. Evaluation of the polymerization reaction of a primer on an M13mp19(+) template by human DNA polymerase α indicated that CNDACTP was incorporated effectively (Km = 0.22 μM) opposite a complementary dGMP in the template strand. CNDACTP competed with the normal substrate, dCTP, for incorporation, and the two nucleotides showed similar substrate efficiencies (Vmax/Km: dCTP = 0.91; CNDACTP = 0.77). Primer extension was potently inhibited by CNDAC triphosphate (Ki = 23 nM); once the analog had been incorporated, further extension was not observed in vitro, suggesting that primers containing a 3′-terminal nucleotide analog were high Km substrates for polymerase α. Thus, the ability of human leukemia cells to effectively accumulate and retain CNDACTP, coupled with the favorable kinetics of competition for incorporation into DNA, and the relatively strong ability of the analog to terminate further extension, are likely to contribute to the cytotoxic action of CNDAC.
KW - CNDAC
KW - DNA polymerase
KW - DNA synthesis
KW - Nucleoside analog
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U2 - 10.1016/S0006-2952(01)00617-7
DO - 10.1016/S0006-2952(01)00617-7
M3 - Article
C2 - 11377379
AN - SCOPUS:0035876975
SN - 0006-2952
VL - 61
SP - 1497
EP - 1507
JO - Biochemical Pharmacology
JF - Biochemical Pharmacology
IS - 12
ER -