Design, synthesis, biological evaluation and X-ray crystal structure of novel classical 6,5,6-tricyclic benzo[4,5]thieno[2,3-d]pyrimidines as dual thymidylate synthase and dihydrofolate reductase inhibitors

DOI

10.1016/j.bmc.2011.03.067

Document Type

Journal Article

Publication Date

6-1-2011

Publication Title

Bioorganic and Medicinal Chemistry

Volume

19

Issue

11

First Page

3585

Last Page

3594

ISSN

9680896

Keywords

DHFR, Dual, Inhibitor, TS

Abstract

Classical antifolates (4-7) with a tricyclic benzo[4,5]thieno[2,3-d] pyrimidine scaffold and a flexible and rigid benzoylglutamate were synthesized as dual thymidylate synthase (TS) and dihydrofolate reductase (DHFR) inhibitors. Oxidative aromatization of ethyl 2-amino-4-methyl-4,5,6,7-tetrahydro-1- benzothiophene-3-carboxylate (±)-9 to ethyl 2-amino-4-methyl-1- benzothiophene-3-carboxylate 10 with 10% Pd/C was a key synthetic step. Compounds with 2-CH3 substituents inhibited human (h) TS (IC 50 = 0.26-0.8 μM), but not hDHFR. Substitution of the 2-CH 3 with a 2-NH2 increases hTS inhibition by more than 10-fold and also affords excellent hDHFR inhibition (IC50 = 0.09-0.1 μM). This study shows that the tricyclic benzo[4,5]thieno[2,3-d]pyrimidine scaffold is highly conducive to single hTS or dual hTS-hDHFR inhibition depending on the 2-position substituents. The X-ray crystal structures of 6 and 7 with hDHFR reveal, for the first time, that tricyclics 6 and 7 bind with the benzo[4,5]thieno[2,3-d]pyrimidine ring in the folate binding mode with the thieno S mimicking the 4-amino of methotrexate. © 2011 Elsevier Ltd. All rights reserved.

Open Access

Green Accepted

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