Classical and nonclassical furo[2,3-d]pyrimidines as novel antifolates: Synthesis and biological activities

Aleem Gangjee, Rajesh Devraj, John J. McGuire, Roy L. Kisliuk, Sherry Queener, Louis R. Barrows

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Classical antifolate analogues containing a novel furo[2,3-d]pyrimidine ring system which include N-[4-[N-[(2,4-diaminofuro[2,3-d] pyrimidin-5-yl)methyl]amino]benzoyl]-L-glutamic acid (1) and its N-9 methyl analogue 2 were synthesized as potential dual inhibitors of thymidylate synthase (TS) and dihydrofolate reductase (DHFR) and as antitumor agents. Four nonclassical antifolates, 2,4-diamino-5-(anilinomethyl)furo[2,3-d]pyrimidmes 3-6 with 3,4,5-trimethoxy, 3,4,5-trichloro, 3,4-dichloro, and 2,5-dimethoxy substituents, respectively, in the phenyl ring, were also synthesized as potential inhibitors of DHFRs including those from Pneumocystis carinii and Toxoplasma gondii, which are organisms responsible for opportunistic infections in AIDS patients. The classical and nonclassical analogues were obtained via nucleophilic displacements of the key intermediate 2,4-diamino-5-(chloromethyl)furo[2,3-d]Pyrimidine with the appropriate (p-aminobenzoyl)-L-glutamate or substituted aniline. The key intermediate was in turn synthesized from 2,4-diamino-6-hydroxypyrimidine and 1,3-dichloroacetone. The final compounds were tested in vitro against rat liver, (recombinant) human, P. carinii, T. gondii, and Lactobacillus casei DHFRs. The classical analogues showed moderate to good DHFR inhibitory activity (IC50 10-6-10-8 M) with the N-CH3 analogue 2 about twice as potent as 1. The nonclassical analogues were inactive with IC50s > 3 × 10-5 M. The classical analogues were also evaluated as inhibitors of TS (L. casei, (recombinant) human and human CCRF-CEM), glycinamide ribonucleotide formyltransferase, and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase and were found to be inactive against these enzymes. The classical analogues (particularly 2) were significantly cytotoxic toward a variety of tumor cell lines in culture. The nonclassical analogues were marginally active. Both classical compounds were good substrates for human folylpolyglutamate synthetase. Further evaluation of the cytotoxicity of 1 and 2 in CCRF-CEM cells and its sublines, having defined mechanisms of methotrexate (MTX) resistance, demonstrated that the analogues utilize the reduced folate/ MTX-transport system and primarily inhibit DHFR and that poly-γ-glutamylation was crucial to their mechanism of action. Protection studies in the FaDu squamous cell carcinoma cell line indicated that inhibition was completely reversed by leucovorin or the combination of thymidine plus hypoxanthine. Furthermore, for compounds 1 and 2, in contrast to MTX, the FaDu cells were better protected by thymidine alone than hypoxanthine alone, suggesting a predominantly antithymidylate effect.

Original languageEnglish
Pages (from-to)1169-1176
Number of pages8
JournalJournal of Medicinal Chemistry
Volume37
Issue number8
StatePublished - 1994

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Folic Acid Antagonists
Pyrimidines
Hydroxymethyl and Formyl Transferases
Bioactivity
Methotrexate
Tetrahydrofolate Dehydrogenase
Hypoxanthine
Lactobacillus casei
Pneumocystis carinii
Thymidine
Glutamic Acid
Toxoplasma
Cells
Ribonucleotides
Thymidylate Synthase
Leucovorin
Cytotoxicity
Folic Acid
Cell culture
Liver

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Gangjee, A., Devraj, R., McGuire, J. J., Kisliuk, R. L., Queener, S., & Barrows, L. R. (1994). Classical and nonclassical furo[2,3-d]pyrimidines as novel antifolates: Synthesis and biological activities. Journal of Medicinal Chemistry, 37(8), 1169-1176.

Classical and nonclassical furo[2,3-d]pyrimidines as novel antifolates : Synthesis and biological activities. / Gangjee, Aleem; Devraj, Rajesh; McGuire, John J.; Kisliuk, Roy L.; Queener, Sherry; Barrows, Louis R.

In: Journal of Medicinal Chemistry, Vol. 37, No. 8, 1994, p. 1169-1176.

Research output: Contribution to journalArticle

Gangjee, A, Devraj, R, McGuire, JJ, Kisliuk, RL, Queener, S & Barrows, LR 1994, 'Classical and nonclassical furo[2,3-d]pyrimidines as novel antifolates: Synthesis and biological activities', Journal of Medicinal Chemistry, vol. 37, no. 8, pp. 1169-1176.
Gangjee, Aleem ; Devraj, Rajesh ; McGuire, John J. ; Kisliuk, Roy L. ; Queener, Sherry ; Barrows, Louis R. / Classical and nonclassical furo[2,3-d]pyrimidines as novel antifolates : Synthesis and biological activities. In: Journal of Medicinal Chemistry. 1994 ; Vol. 37, No. 8. pp. 1169-1176.
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T1 - Classical and nonclassical furo[2,3-d]pyrimidines as novel antifolates

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