Mechanism of cytostatic action of novel 5-(thien-2-yl)- and 5-(furan-2-yl)-substituted pyrimidine nucleoside analogues against tumor cells transfected by the thymidine kinase gene of herpes simplex virus.

Abstract

Several novel 5-substituted 2'-deoxyuridine (dUrd) analogues were evaluated as substrates for highly purified herpes simplex virus type 1 (HSV-1)-encoded thymidine kinase (TK) derived from HSV-1 TK gene-transfected murine mammary carcinoma FM3A cells, and human platelet thymidine (dThd) phosphorylase. The Ki of 5-(furan-2-yl)-dUrd, 5-(thien-2-yl)-dUrd and 5-(thien-2-yl)-dCyd for HSV-1 TK was 0.94, 0.71, and 1.32 microM, respectively. Inhibition was competitive with respect to the natural substrate dThd. With dCyd as substrate, the Ki of 5-(thien-2-yl)-dCyd for HSV-1 TK was 4.5 microM (Ki/Km = 0.17). In striking contrast with (E)-5-(2-bromovinyl)-dUrd, the 5-(thien-2-yl)- and 5-(furan-2-yl)-dUrd derivatives were not substrates for human dThd phosphorylase. 5-(Thien-2-yl)-dUrd, 5-(furan-2-yl)-dUrd and 5-(thien-2-yl)-dCyd were at least 100-fold more cytostatic to the HSV TK gene-transfected FM3A tumor cells than wild-type FM3A/0 cells. The viral TK expressed in the HSV-1 TK gene-transfected tumor cells merely acts as an activating enzyme, whereas thymidylate synthase serves as the target enzyme for the cytostatic action of the compounds. The novel 5-substituted dUrd analogues should be further pursued as candidate drugs in the treatment of HSV TK gene-transfected tumors.