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10-formyl-5,10-dideaza-acyclic-5,6,7,8-tetrahydrofolic acid (10-formyl-DDACTHF) a potent cytotoxic agent acting by selective inhibition of human GAR Tfase and the de novo purine biosynthetic pathway

10-formyl-5,10-dideaza-acyclic-5,6,7,8-tetrahydrofolic acid (10-formyl-DDACTHF) a potent cytotoxic agent acting by selective inhibition of human GAR Tfase and the de novo purine biosynthetic pathway
10-formyl-5,10-dideaza-acyclic-5,6,7,8-tetrahydrofolic acid (10-formyl-DDACTHF) a potent cytotoxic agent acting by selective inhibition of human GAR Tfase and the de novo purine biosynthetic pathway
The synthesis of 10-formyl-DDACTHF (3) as a potential inhibitor of glycinamide ribonucleotide transformylase (GAR Tfase) and aminoimidazole carboxamide ribonucleotide transformylase (AICAR Tfase) is reported. Aldehyde 3, the corresponding gamma- and alpha-pentaglutamates 21 and 25 and related agents were evaluated for inhibition of folate-dependent enzymes including GAR Tfase and AICAR Tfase. The inhibitors were found to exhibit potent cytotoxic activity (CCRF-CEM IC(50) for 3=60nM) that exceeded their enzyme inhibition potency [K(i) (3)=6 and 1 microM for Escherichia coli GAR and human AICAR Tfase, respectively]. Cytotoxicity rescue by medium purines, but not pyrimidines, indicated that the potent cytotoxic activity is derived from selective purine biosynthesis inhibition and rescue by AICAR monophosphate established that the activity is derived preferentially from GAR versus AICAR Tfase inhibition. The potent cytotoxic compounds including aldehyde 3 lost activity against CCRF-CEM cell lines deficient in the reduced folate carrier (CCRF-CEM/MTX) or folylpolyglutamate synthase (CCRF-CEM/FPGS(-)) establishing that their potent activity requires both reduced folate carrier transport and polyglutamation. Unexpectedly, the pentaglutamates displayed surprisingly similar K(i)'s versus E. coli GAR Tfase and only modestly enhanced K(i)'s versus human AICAR Tfase. On the surface this initially suggested that the potent cytotoxic activity of 3 and related compounds might be due simply to preferential intracellular accumulation of the inhibitors derived from effective transport and polyglutamation (i.e., ca. 100-fold higher intracellular concentrations). However, a subsequent examination of the inhibitors against recombinant human GAR Tfase revealed they and the corresponding gamma-pentaglutamates were unexpectedly much more potent against the human versus E. coli enzyme (K(i) for 3, 14nM against rhGAR Tfase versus 6 microM against E. coli GAR Tfase) which also accounts for their exceptional cytotoxic potency.
antineoplastic agents, chemical synthesis/pharmacology carrier proteins/physiology cell division/drug effects enzyme inhibitors/chemical synthesis/pharmacology humans hydroxymethyl and formyl transferases, antagonists & inhibitors peptide synthases/physiology phosphoribosylaminoimidazolecarboxamide formyltransferase phosphoribosylglycinamide formyltransferase purines/antagonists & inhibitors, biosynthesis *receptors, cell surface structure-activity relationship tetrahydrofolates, chemical synthesis tumor cells, cultured
0968-0896
2739-2749
Marsilje, Thomas H.
2d64bf3a-143b-4a00-8b9c-825beceb810e
Labroli, Marc A.
aa05ca91-73ec-49bf-b2b7-7542945efa56
Hedrick, Michael P.
ba690321-bfda-496d-98de-c3d383497035
Jin, Qing
3597a036-15bf-4ad2-a5b5-fe909811d2aa
Desharnais, Joel
f4d5b1db-e99b-4487-b37a-f64f24c7914a
Baker, Stephen J.
98c076ff-72c9-4ee2-a027-e58560fdfd43
Gooljarsingh, Lata T.
e63899ea-fc7a-4adf-aaf7-b9921b37d1fc
Ramcharan, Joseph
53296e87-e26b-485c-8f13-f31642a58632
Tavassoli, Ali
d561cf8f-2669-46b5-b6e1-2016c85d63b2
Zhang, Yan
8552a39d-0c4d-4712-8ad8-d0d5cc9d1779
Wilson, Ian A.
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Beardsley, G. Peter
85b7ede0-bcc4-4d31-980e-a0aee69c842e
Benkovic, Stephen J.
f85498a5-27fc-4a22-9069-ff61a4297520
Boger, Dale L.
d81e0270-0561-47df-8904-85718bd462ff
Marsilje, Thomas H.
2d64bf3a-143b-4a00-8b9c-825beceb810e
Labroli, Marc A.
aa05ca91-73ec-49bf-b2b7-7542945efa56
Hedrick, Michael P.
ba690321-bfda-496d-98de-c3d383497035
Jin, Qing
3597a036-15bf-4ad2-a5b5-fe909811d2aa
Desharnais, Joel
f4d5b1db-e99b-4487-b37a-f64f24c7914a
Baker, Stephen J.
98c076ff-72c9-4ee2-a027-e58560fdfd43
Gooljarsingh, Lata T.
e63899ea-fc7a-4adf-aaf7-b9921b37d1fc
Ramcharan, Joseph
53296e87-e26b-485c-8f13-f31642a58632
Tavassoli, Ali
d561cf8f-2669-46b5-b6e1-2016c85d63b2
Zhang, Yan
8552a39d-0c4d-4712-8ad8-d0d5cc9d1779
Wilson, Ian A.
7865d500-d638-4a67-ad6d-fefad0ae83bb
Beardsley, G. Peter
85b7ede0-bcc4-4d31-980e-a0aee69c842e
Benkovic, Stephen J.
f85498a5-27fc-4a22-9069-ff61a4297520
Boger, Dale L.
d81e0270-0561-47df-8904-85718bd462ff

Marsilje, Thomas H., Labroli, Marc A., Hedrick, Michael P., Jin, Qing, Desharnais, Joel, Baker, Stephen J., Gooljarsingh, Lata T., Ramcharan, Joseph, Tavassoli, Ali, Zhang, Yan, Wilson, Ian A., Beardsley, G. Peter, Benkovic, Stephen J. and Boger, Dale L. (2002) 10-formyl-5,10-dideaza-acyclic-5,6,7,8-tetrahydrofolic acid (10-formyl-DDACTHF) a potent cytotoxic agent acting by selective inhibition of human GAR Tfase and the de novo purine biosynthetic pathway. Bioorganic & Medicinal Chemistry, 10 (8), 2739-2749. (doi:10.1016/S0968-0896(02)00102-5).

Record type: Article

Abstract

The synthesis of 10-formyl-DDACTHF (3) as a potential inhibitor of glycinamide ribonucleotide transformylase (GAR Tfase) and aminoimidazole carboxamide ribonucleotide transformylase (AICAR Tfase) is reported. Aldehyde 3, the corresponding gamma- and alpha-pentaglutamates 21 and 25 and related agents were evaluated for inhibition of folate-dependent enzymes including GAR Tfase and AICAR Tfase. The inhibitors were found to exhibit potent cytotoxic activity (CCRF-CEM IC(50) for 3=60nM) that exceeded their enzyme inhibition potency [K(i) (3)=6 and 1 microM for Escherichia coli GAR and human AICAR Tfase, respectively]. Cytotoxicity rescue by medium purines, but not pyrimidines, indicated that the potent cytotoxic activity is derived from selective purine biosynthesis inhibition and rescue by AICAR monophosphate established that the activity is derived preferentially from GAR versus AICAR Tfase inhibition. The potent cytotoxic compounds including aldehyde 3 lost activity against CCRF-CEM cell lines deficient in the reduced folate carrier (CCRF-CEM/MTX) or folylpolyglutamate synthase (CCRF-CEM/FPGS(-)) establishing that their potent activity requires both reduced folate carrier transport and polyglutamation. Unexpectedly, the pentaglutamates displayed surprisingly similar K(i)'s versus E. coli GAR Tfase and only modestly enhanced K(i)'s versus human AICAR Tfase. On the surface this initially suggested that the potent cytotoxic activity of 3 and related compounds might be due simply to preferential intracellular accumulation of the inhibitors derived from effective transport and polyglutamation (i.e., ca. 100-fold higher intracellular concentrations). However, a subsequent examination of the inhibitors against recombinant human GAR Tfase revealed they and the corresponding gamma-pentaglutamates were unexpectedly much more potent against the human versus E. coli enzyme (K(i) for 3, 14nM against rhGAR Tfase versus 6 microM against E. coli GAR Tfase) which also accounts for their exceptional cytotoxic potency.

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Published date: August 2002
Keywords: antineoplastic agents, chemical synthesis/pharmacology carrier proteins/physiology cell division/drug effects enzyme inhibitors/chemical synthesis/pharmacology humans hydroxymethyl and formyl transferases, antagonists & inhibitors peptide synthases/physiology phosphoribosylaminoimidazolecarboxamide formyltransferase phosphoribosylglycinamide formyltransferase purines/antagonists & inhibitors, biosynthesis *receptors, cell surface structure-activity relationship tetrahydrofolates, chemical synthesis tumor cells, cultured
Organisations: Chemistry

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Local EPrints ID: 45316
URI: http://eprints.soton.ac.uk/id/eprint/45316
ISSN: 0968-0896
PURE UUID: ab4df612-31bb-4298-81e2-2d3c16e9d06d
ORCID for Ali Tavassoli: ORCID iD orcid.org/0000-0002-7420-5063

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Date deposited: 21 Mar 2007
Last modified: 09 Jan 2022 03:22

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Contributors

Author: Thomas H. Marsilje
Author: Marc A. Labroli
Author: Michael P. Hedrick
Author: Qing Jin
Author: Joel Desharnais
Author: Stephen J. Baker
Author: Lata T. Gooljarsingh
Author: Joseph Ramcharan
Author: Ali Tavassoli ORCID iD
Author: Yan Zhang
Author: Ian A. Wilson
Author: G. Peter Beardsley
Author: Stephen J. Benkovic
Author: Dale L. Boger

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