The methods of the present invention allow for the measurement of
ribonucleotide reductase (RR) activity, an important
enzyme in the de novo
DNA synthesis pathway.
Ribonucleotide reductase converts all four ribonucleotides to their deoxy form and is a rate-controlling step in this pathway. Biosynthetic pathways of deoxyribonucleotides (dN) have received considerable attention in the context of anti-proliferative
chemotherapy. Inhibitors of various steps in dN
biosynthesis, including inhibitors of RR are among the most useful chemotherapeutic agents in
cancer, viral infections, and other therapeutic uses.
DNA synthesis from the dN salvage pathway is also an important component to
DNA replication. The relative contributions from RR vs. salvage pathways are critical to the actions and effectiveness of chemotherapeutic agents that act on
nucleoside metabolic pathways. Until now, however, it has not been possible to study these metabolic processes
in vivo. Disclosed within are methods of measuring RR activity
in vivo and
in vitro which find use, among other things, in
drug discovery, development, and approval.