Combination studies with 3′-azido-3′-deoxythymidine (AZT) plus ICI D1694. Cytotoxic and biochemical effects

Josie Pressacco, Charles Erlichman

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The cytotoxicity of 3′-azido-3′-deoxythymidine (AZT), a thymidine analogue, and ICI D1694, a folate-based thymidylate synthase (TS) inhibitor, was examined individually and in combination in two human tumor cell lines, MGH-U1 bladder cancer and HCT-8 colon cancer cells, grown as a monolayer culture with and without thymidine (TdR). In addition, TS inhibition, [3H]AZT incorporation into DNA, [3H]AZT-MP (monophosphate) production, and DNA double-strand breaks were measured. Twenty-four hour exposure of AZT at 0.5, 5, 50 and 500 μM was not cytotoxic to MGH-U1 or HCT-8 cells in a colony-forming assay. ICI D1694 cytotoxicity increased with drug concentration, and the ic50 and ic90, respectively, were 0.0064 and 0.01 μM in MGH-U1 cells and 0.009 and 0.018 μM in HCT-8 cells. TdR in concentrations of 0.1 to 1.0 μM did not affect ICI D1694 cytotoxicity in either cell line. AZT at 5, 50 or 500 μM increased ICI D1694 cell kill. The ic50 and ic50 for MGH-U1 were 0.0037 and 0.0075 μM for 50 μM AZT combined with ICI D1694. The ic90 for HCT-8 were 0.0075 and 0.015 μM for 50 μM AZT plus ICI D1694. The incorporation of [3H]AZT into DNA increased with increasing concentrations of ICI D1694. Concentrations producing an ic50 and ic90 of ICI D1694, respectively, increased incorporation of [3H]AZT into DNA by 319 and 569% in MHG-U1, and 243 and 400% in HCT-8 cells. The formation of [3H]AZT-MP paralleled the increase in [3H]AZT incorporated into DNA. AZT, ICI D1694 and the combination of AZT and ICI D1694 caused DNA double-strand breaks, with the combination of these agents being additive. CFU-GM survival, exposed to drug concentrations, as those used in the tumor cell lines, revealed that the therapeutic index was greater for AZT plus ICI D1694 than for ICI D1694 alone. These findings suggest that AZT plus ICI D1694 may increase antitumor effect with minimal myelosuppression. We conclude that AZT increases the cytotoxicity of ICI D1694 with increasing AZT incorporation into DNA.

Original languageEnglish (US)
Pages (from-to)1989-1997
Number of pages9
JournalBiochemical Pharmacology
Volume46
Issue number11
DOIs
StatePublished - Dec 3 1993
Externally publishedYes

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Zidovudine
Cytotoxicity
DNA
Inhibitory Concentration 50
Cells
Thymidylate Synthase
Double-Stranded DNA Breaks
raltitrexed
Tumor Cell Line
Thymidine
Tumors
Granulocyte-Macrophage Progenitor Cells
Folic Acid
Cell culture
Urinary Bladder Neoplasms
Pharmaceutical Preparations
Colonic Neoplasms
Monolayers
Assays

ASJC Scopus subject areas

  • Pharmacology

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Combination studies with 3′-azido-3′-deoxythymidine (AZT) plus ICI D1694. Cytotoxic and biochemical effects. / Pressacco, Josie; Erlichman, Charles.

In: Biochemical Pharmacology, Vol. 46, No. 11, 03.12.1993, p. 1989-1997.

Research output: Contribution to journalArticle

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abstract = "The cytotoxicity of 3′-azido-3′-deoxythymidine (AZT), a thymidine analogue, and ICI D1694, a folate-based thymidylate synthase (TS) inhibitor, was examined individually and in combination in two human tumor cell lines, MGH-U1 bladder cancer and HCT-8 colon cancer cells, grown as a monolayer culture with and without thymidine (TdR). In addition, TS inhibition, [3H]AZT incorporation into DNA, [3H]AZT-MP (monophosphate) production, and DNA double-strand breaks were measured. Twenty-four hour exposure of AZT at 0.5, 5, 50 and 500 μM was not cytotoxic to MGH-U1 or HCT-8 cells in a colony-forming assay. ICI D1694 cytotoxicity increased with drug concentration, and the ic50 and ic90, respectively, were 0.0064 and 0.01 μM in MGH-U1 cells and 0.009 and 0.018 μM in HCT-8 cells. TdR in concentrations of 0.1 to 1.0 μM did not affect ICI D1694 cytotoxicity in either cell line. AZT at 5, 50 or 500 μM increased ICI D1694 cell kill. The ic50 and ic50 for MGH-U1 were 0.0037 and 0.0075 μM for 50 μM AZT combined with ICI D1694. The ic90 for HCT-8 were 0.0075 and 0.015 μM for 50 μM AZT plus ICI D1694. The incorporation of [3H]AZT into DNA increased with increasing concentrations of ICI D1694. Concentrations producing an ic50 and ic90 of ICI D1694, respectively, increased incorporation of [3H]AZT into DNA by 319 and 569{\%} in MHG-U1, and 243 and 400{\%} in HCT-8 cells. The formation of [3H]AZT-MP paralleled the increase in [3H]AZT incorporated into DNA. AZT, ICI D1694 and the combination of AZT and ICI D1694 caused DNA double-strand breaks, with the combination of these agents being additive. CFU-GM survival, exposed to drug concentrations, as those used in the tumor cell lines, revealed that the therapeutic index was greater for AZT plus ICI D1694 than for ICI D1694 alone. These findings suggest that AZT plus ICI D1694 may increase antitumor effect with minimal myelosuppression. We conclude that AZT increases the cytotoxicity of ICI D1694 with increasing AZT incorporation into DNA.",
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N2 - The cytotoxicity of 3′-azido-3′-deoxythymidine (AZT), a thymidine analogue, and ICI D1694, a folate-based thymidylate synthase (TS) inhibitor, was examined individually and in combination in two human tumor cell lines, MGH-U1 bladder cancer and HCT-8 colon cancer cells, grown as a monolayer culture with and without thymidine (TdR). In addition, TS inhibition, [3H]AZT incorporation into DNA, [3H]AZT-MP (monophosphate) production, and DNA double-strand breaks were measured. Twenty-four hour exposure of AZT at 0.5, 5, 50 and 500 μM was not cytotoxic to MGH-U1 or HCT-8 cells in a colony-forming assay. ICI D1694 cytotoxicity increased with drug concentration, and the ic50 and ic90, respectively, were 0.0064 and 0.01 μM in MGH-U1 cells and 0.009 and 0.018 μM in HCT-8 cells. TdR in concentrations of 0.1 to 1.0 μM did not affect ICI D1694 cytotoxicity in either cell line. AZT at 5, 50 or 500 μM increased ICI D1694 cell kill. The ic50 and ic50 for MGH-U1 were 0.0037 and 0.0075 μM for 50 μM AZT combined with ICI D1694. The ic90 for HCT-8 were 0.0075 and 0.015 μM for 50 μM AZT plus ICI D1694. The incorporation of [3H]AZT into DNA increased with increasing concentrations of ICI D1694. Concentrations producing an ic50 and ic90 of ICI D1694, respectively, increased incorporation of [3H]AZT into DNA by 319 and 569% in MHG-U1, and 243 and 400% in HCT-8 cells. The formation of [3H]AZT-MP paralleled the increase in [3H]AZT incorporated into DNA. AZT, ICI D1694 and the combination of AZT and ICI D1694 caused DNA double-strand breaks, with the combination of these agents being additive. CFU-GM survival, exposed to drug concentrations, as those used in the tumor cell lines, revealed that the therapeutic index was greater for AZT plus ICI D1694 than for ICI D1694 alone. These findings suggest that AZT plus ICI D1694 may increase antitumor effect with minimal myelosuppression. We conclude that AZT increases the cytotoxicity of ICI D1694 with increasing AZT incorporation into DNA.

AB - The cytotoxicity of 3′-azido-3′-deoxythymidine (AZT), a thymidine analogue, and ICI D1694, a folate-based thymidylate synthase (TS) inhibitor, was examined individually and in combination in two human tumor cell lines, MGH-U1 bladder cancer and HCT-8 colon cancer cells, grown as a monolayer culture with and without thymidine (TdR). In addition, TS inhibition, [3H]AZT incorporation into DNA, [3H]AZT-MP (monophosphate) production, and DNA double-strand breaks were measured. Twenty-four hour exposure of AZT at 0.5, 5, 50 and 500 μM was not cytotoxic to MGH-U1 or HCT-8 cells in a colony-forming assay. ICI D1694 cytotoxicity increased with drug concentration, and the ic50 and ic90, respectively, were 0.0064 and 0.01 μM in MGH-U1 cells and 0.009 and 0.018 μM in HCT-8 cells. TdR in concentrations of 0.1 to 1.0 μM did not affect ICI D1694 cytotoxicity in either cell line. AZT at 5, 50 or 500 μM increased ICI D1694 cell kill. The ic50 and ic50 for MGH-U1 were 0.0037 and 0.0075 μM for 50 μM AZT combined with ICI D1694. The ic90 for HCT-8 were 0.0075 and 0.015 μM for 50 μM AZT plus ICI D1694. The incorporation of [3H]AZT into DNA increased with increasing concentrations of ICI D1694. Concentrations producing an ic50 and ic90 of ICI D1694, respectively, increased incorporation of [3H]AZT into DNA by 319 and 569% in MHG-U1, and 243 and 400% in HCT-8 cells. The formation of [3H]AZT-MP paralleled the increase in [3H]AZT incorporated into DNA. AZT, ICI D1694 and the combination of AZT and ICI D1694 caused DNA double-strand breaks, with the combination of these agents being additive. CFU-GM survival, exposed to drug concentrations, as those used in the tumor cell lines, revealed that the therapeutic index was greater for AZT plus ICI D1694 than for ICI D1694 alone. These findings suggest that AZT plus ICI D1694 may increase antitumor effect with minimal myelosuppression. We conclude that AZT increases the cytotoxicity of ICI D1694 with increasing AZT incorporation into DNA.

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