Cellular transformation of the investigational new anticancer drug NB1011, a phosphoramidate of 5-(2-bromovinyl)-2'-deoxyuridine, results in modification of cellular proteins not DNA.
NB1011 [E-5-(2-bromovinyl)-2'-deoxyuridine-5'-(L-methylalaninyl)-phenylphosphoramidate], a phosphoramidate prodrug of E-5-(2-bromovinyl)-2'-deoxyuridine-5'-monophosphate (BVdUMP), is an investigational new anticancer drug. NB1011 targets thymidylate synthase (TS), which catalyzes the transformation of BVdUMP into cytotoxic reaction products. Due to the elevated levels of TS expression in tumor cells compared to normal cells, these cytotoxic products are preferentially generated inside tumor cells, and, as expected, NB1011 is more toxic to cells with higher levels of TS expression. Therefore, NB1011 therapy should kill tumor cells without severely damaging normal cells. Radiolabeled NB1011 was used to determine the intracellular fate of NB1011 reaction products and, possibly, the mechanism of action of this investigational new drug. We found significant incorporation of the radiolabel into cellular macromolecules. In contrast to our expectations that NB1011 product(s) would be incorporated into DNA, we discovered that cellular proteins were the labeled macromolecular fraction. Herein, we report that the intracellular transformation of NB1011 involves formation of the corresponding monophosphate, TS-dependent transformation into highly reactive intermediates, and subsequent incorporation into cellular proteins. TS itself appears to escape irreversible inactivation. Our data suggest that protein modification not DNA incorporation accounts for the therapeutic effect of NB1011. The proposed mechanism is rather unexpected for a nucleotide analogue and could lead to the discovery of new cellular protein targets for future drug design.
Sergeeva
Maria V
MV
NewBiotics, Inc., 4939 Directors Place, San Diego, CA 92121, USA. msergeeva@newbiotics.com
Cathers
Brian E
BE
D000970:Antineoplastic Agents / Q000009:adverse effects* / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000009:adverse effects* / Q000031:analogs & derivatives / Q000378:metabolism; D002455:Cell Division; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced*; D002586:Cesium / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D004273:DNA, Neoplasm / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D011499:Protein Processing, Post-Translational; D013940:Thymidylate Synthase / Q000378:metabolism; D014238:Trichloroacetic Acid / Q000737:chemistry; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D002712:Chlorides; D004273:DNA, Neoplasm; C421011:NB1011; D002586:Cesium; D014238:Trichloroacetic Acid; D013940:Thymidylate Synthase; D001973:Bromodeoxyuridine; C028019:cesium chloride