Nonclassical 2,4-diamino-5-aryl-6-ethylpyrimidine antifolates: Activity as inhibitors of dihydrofolate reductase from Pneumocystis carinii and Toxoplasma gondii and as antitumor agents

Claire Robson, Michelle A. Meek, Jan Dierk Grunwaldt, Peter A. Lambert, Sherry Queener, Dirk Schmidt, Roger J. Griffin

Research output: Contribution to journalArticle

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Abstract

Twelve novel 2,4-diamino-5-(4'-benzylamino)- and 2,4-diamino-5-[4'-(N- methylbenzylamino)-3'-nitrophenyl]-6-ethylpyrimidines bearing 4-substituents on the benzylamino or N-methylbenzylamino aryl ring were synthesized and evaluated as nonclassical inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase (DHFR). Compounds were prepared by reaction of 2,4-diamino-5-(4'-chloro-3'-nitrophenyl)-(8) or 2,4-diamino-5-(4'-fluoro-3'- nitrophenyl)-6-ethylpyrimidine (15) with the appropriate 4-substituted (CO 2H, CO 2Me, SO 2NH 2, dioxolan-2-yl, CHO, dimethyloxazolin-2-yl) benzylamine or N-methylbenzylamine derivative. Compounds 25-29 were synthesized from 2,4-diamino-5-{4'-[N-(4''-carboxybenzyl)amino]-3'- nitrophenyl}-6-ethylpyrimidine (10) and the corresponding amine (NH 3, MeNH 2, Me 2NH, piperidine, diethyl L-glutamate) via isobutyl mixed anhydride coupling; hydrolysis of the diethyl L-glutamate 29 afforded the L-glutamate analogue 30. The compounds exhibited potent inhibitory activity against T. gondii (IC 50 values 0.0018-0.14 μM) and rat liver (IC 50 values 0.0029- 0.27 μM) DHFR, with a 4-substituent invariably enhancing binding to both enzymes relative to the unsubstituted benzoprim (5) or methylbenzoprim (6). Modest selectivity for T. gondii enzyme was observed with several analogues, whereas all of the compounds were relatively weak inhibitors of P. carinii DHFR and exhibited no selectivity. Selected analogues were evaluated for in vivo antitumor activity against the methotrexate-resistant M5076 murine reticulosarcoma, with 2,4-diamino-5-{4'-[N-[4''-(N'- methylcarbamoyl)benzyl]-N-methylamino]-3'-nitrophenyl}-6-ethylpyrimidine (14) (K(i) for rat liver DHFR = 0.000 35 ± 0.000 29 nM) combining significant antitumor activity with minimal toxicity.

Original languageEnglish
Pages (from-to)3040-3048
Number of pages9
JournalJournal of Medicinal Chemistry
Volume40
Issue number19
DOIs
StatePublished - 1997

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Folic Acid Antagonists
Pneumocystis carinii
Tetrahydrofolate Dehydrogenase
Toxoplasma
Antineoplastic Agents
Glutamic Acid
Carbon Monoxide
Liver
Rats
Bearings (structural)
Anhydrides
Enzymes
Methotrexate
Non-Hodgkin's Lymphoma
Amines
Toxicity
Hydrolysis
Derivatives

ASJC Scopus subject areas

  • Organic Chemistry

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Nonclassical 2,4-diamino-5-aryl-6-ethylpyrimidine antifolates : Activity as inhibitors of dihydrofolate reductase from Pneumocystis carinii and Toxoplasma gondii and as antitumor agents. / Robson, Claire; Meek, Michelle A.; Grunwaldt, Jan Dierk; Lambert, Peter A.; Queener, Sherry; Schmidt, Dirk; Griffin, Roger J.

In: Journal of Medicinal Chemistry, Vol. 40, No. 19, 1997, p. 3040-3048.

Research output: Contribution to journalArticle

Robson, Claire ; Meek, Michelle A. ; Grunwaldt, Jan Dierk ; Lambert, Peter A. ; Queener, Sherry ; Schmidt, Dirk ; Griffin, Roger J. / Nonclassical 2,4-diamino-5-aryl-6-ethylpyrimidine antifolates : Activity as inhibitors of dihydrofolate reductase from Pneumocystis carinii and Toxoplasma gondii and as antitumor agents. In: Journal of Medicinal Chemistry. 1997 ; Vol. 40, No. 19. pp. 3040-3048.
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abstract = "Twelve novel 2,4-diamino-5-(4'-benzylamino)- and 2,4-diamino-5-[4'-(N- methylbenzylamino)-3'-nitrophenyl]-6-ethylpyrimidines bearing 4-substituents on the benzylamino or N-methylbenzylamino aryl ring were synthesized and evaluated as nonclassical inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase (DHFR). Compounds were prepared by reaction of 2,4-diamino-5-(4'-chloro-3'-nitrophenyl)-(8) or 2,4-diamino-5-(4'-fluoro-3'- nitrophenyl)-6-ethylpyrimidine (15) with the appropriate 4-substituted (CO 2H, CO 2Me, SO 2NH 2, dioxolan-2-yl, CHO, dimethyloxazolin-2-yl) benzylamine or N-methylbenzylamine derivative. Compounds 25-29 were synthesized from 2,4-diamino-5-{4'-[N-(4''-carboxybenzyl)amino]-3'- nitrophenyl}-6-ethylpyrimidine (10) and the corresponding amine (NH 3, MeNH 2, Me 2NH, piperidine, diethyl L-glutamate) via isobutyl mixed anhydride coupling; hydrolysis of the diethyl L-glutamate 29 afforded the L-glutamate analogue 30. The compounds exhibited potent inhibitory activity against T. gondii (IC 50 values 0.0018-0.14 μM) and rat liver (IC 50 values 0.0029- 0.27 μM) DHFR, with a 4-substituent invariably enhancing binding to both enzymes relative to the unsubstituted benzoprim (5) or methylbenzoprim (6). Modest selectivity for T. gondii enzyme was observed with several analogues, whereas all of the compounds were relatively weak inhibitors of P. carinii DHFR and exhibited no selectivity. Selected analogues were evaluated for in vivo antitumor activity against the methotrexate-resistant M5076 murine reticulosarcoma, with 2,4-diamino-5-{4'-[N-[4''-(N'- methylcarbamoyl)benzyl]-N-methylamino]-3'-nitrophenyl}-6-ethylpyrimidine (14) (K(i) for rat liver DHFR = 0.000 35 ± 0.000 29 nM) combining significant antitumor activity with minimal toxicity.",
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AU - Grunwaldt, Jan Dierk

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AU - Queener, Sherry

AU - Schmidt, Dirk

AU - Griffin, Roger J.

PY - 1997

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N2 - Twelve novel 2,4-diamino-5-(4'-benzylamino)- and 2,4-diamino-5-[4'-(N- methylbenzylamino)-3'-nitrophenyl]-6-ethylpyrimidines bearing 4-substituents on the benzylamino or N-methylbenzylamino aryl ring were synthesized and evaluated as nonclassical inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase (DHFR). Compounds were prepared by reaction of 2,4-diamino-5-(4'-chloro-3'-nitrophenyl)-(8) or 2,4-diamino-5-(4'-fluoro-3'- nitrophenyl)-6-ethylpyrimidine (15) with the appropriate 4-substituted (CO 2H, CO 2Me, SO 2NH 2, dioxolan-2-yl, CHO, dimethyloxazolin-2-yl) benzylamine or N-methylbenzylamine derivative. Compounds 25-29 were synthesized from 2,4-diamino-5-{4'-[N-(4''-carboxybenzyl)amino]-3'- nitrophenyl}-6-ethylpyrimidine (10) and the corresponding amine (NH 3, MeNH 2, Me 2NH, piperidine, diethyl L-glutamate) via isobutyl mixed anhydride coupling; hydrolysis of the diethyl L-glutamate 29 afforded the L-glutamate analogue 30. The compounds exhibited potent inhibitory activity against T. gondii (IC 50 values 0.0018-0.14 μM) and rat liver (IC 50 values 0.0029- 0.27 μM) DHFR, with a 4-substituent invariably enhancing binding to both enzymes relative to the unsubstituted benzoprim (5) or methylbenzoprim (6). Modest selectivity for T. gondii enzyme was observed with several analogues, whereas all of the compounds were relatively weak inhibitors of P. carinii DHFR and exhibited no selectivity. Selected analogues were evaluated for in vivo antitumor activity against the methotrexate-resistant M5076 murine reticulosarcoma, with 2,4-diamino-5-{4'-[N-[4''-(N'- methylcarbamoyl)benzyl]-N-methylamino]-3'-nitrophenyl}-6-ethylpyrimidine (14) (K(i) for rat liver DHFR = 0.000 35 ± 0.000 29 nM) combining significant antitumor activity with minimal toxicity.

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