The goal of this proposal is to exploit different NMR techniques in order to elucidate several pharmacological properties of a novel anticancer antimetabolite, (E) 2'fluoromethylene-2'-deoxycytidine. 19F NMR imaging and spectroscopy are relatively new approaches that we will utilize to measure in vivo, metabolism and distribution of this deoxycytidine analog. There is a real need for non-invasive assessment of tissue pharmacokinetics and metabolism of anticancer drugs as a way to establish the effectiveness of a drug and to selectively enhance the antineoplastic activity. In vivo studies in a murine tumor model will be conducted in order to improve the necessary techniques and to establish the performance limits of 19F NMR methods for monitoring drug distribution and metabolism in neoplastic and normal tissues. Similar deoxy-cytidine analogs have been shown to be misincorporated into DNA leading to critical lesions of the primary structure of the nucleic acid. Consequently, structural alterations of DNA that lead to changes in the physico-chemical and biochemical properties of the nucleic acid need to be analyzed in order to understand the recognition mechanisms by DNA repair enzymes and how the DNA damage affects both DNA replication and transcription. We will focus on the use of 1H NMR, 19F-NMR and 31P NMR to describe structural distorsions caused by the misincorporation of this antimetabolite in a DNA duplex.
