title: Nucleosides with a twist. Can fixed forms of sugar ring pucker influence biological activity in nucleosides and oligonucleotides?
abstract: The sugar moiety of nucleosides in solution is known to exist in a rapid dynamic equilibrium between extreme Northern and Southern conformations as defined in the pseudorotational cycle. In the present work, we describe how the bicyclo[3.1.0]hexane template fixes the ring pucker of 2'-deoxy-methanocarba-nucleosides 1-5 and 12 to values corresponding to either one of these two extreme conformations that are typical of nucleosides. The syntheses of the fixed Northern conformers 1-5 were performed by Mitsunobu coupling of the heterocyclic bases with the chiral carbocyclic alcohol 6 [(1R,2S,4R,5S)-1-[(benzyloxy)methyl]-2-(tert-butyloxy)-4-hydrox ybicyclo[3.1.0]hexane], while the synthesis of the Southern conformer, (S)-methanocarba-T (12), was reported earlier. Carbocyclic thymidine (carba-T, 13) was used as a reference, flexible carbocyclic nucleoside. Antiviral evaluation of these compounds revealed a very potent antiherpetic activity associated with the Northern thymidine analogue 2, which was more powerful than the reference standard acyclovir against both HSV-1 and HSV-2. (N)-Methanocarba-T (2) was further evaluated as a component of a short oligodeoxynucleotide (ODN) phosphorothioate (5'-CTTCATTTTTTCTTC-3') where all thymidines were replaced by 2. The expected thermodynamic stability resulting from the preorganization of the pseudosugar rings into a Northern conformation, typical of A-DNA, was evident by the increase in Tm of the corresponding DNA/RNA heteroduplex. However, the rigid A-tract ODN caused loss of RNase H recruitment. A detailed conformational analysis of (N)-methanocarba-T (2) and (S)-methanocarba-T (12), as representative examples of conformationally rigid pseudorotational antipodes, revealed that in addition to their different forms of ring pucker, (S)-methanocarba-T appears to be a rather stiff molecule with fewer low-energy conformational states available compared to (N)-methanocarba-T. The syn/anti-energy barrier for these nucleoside analogues is 5-6 kcal/mol higher than for common nucleosides.
authors.affiliation: Laboratory of Medicinal Chemistry, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: V E
authors.initials: VE
authors.lastname: Marquez
authors.forename: M A
authors.initials: MA
authors.lastname: Siddiqui
authors.forename: A
authors.initials: A
authors.lastname: Ezzitouni
authors.forename: P
authors.initials: P
authors.lastname: Russ
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: R W
authors.initials: RW
authors.lastname: Wagner
authors.forename: M D
authors.initials: MD
authors.lastname: Matteucci
mesh_terms: D000212:Acyclovir / Q000494:pharmacology; D016375:Antisense Elements (Genetics) / Q000737:chemistry; D000998:Antiviral Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D001483:Base Sequence; D003588:Cytopathogenic Effect, Viral; D004355:Drug Stability; D018259:Herpesvirus 1, Human / Q000187:drug effects; D018258:Herpesvirus 2, Human / Q000187:drug effects; D006801:Humans; D006860:Hydrogen Bonding; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry; D009705:Nucleosides / Q000737:chemistry* / Q000494:pharmacology; D009841:Oligonucleotides / Q000737:chemistry* / Q000494:pharmacology; D016914:Ribonuclease H / Q000378:metabolism; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D010948:Viral Plaque Assay
chemical_list: D016375:Antisense Elements (Genetics); D000998:Antiviral Agents; D009692:Nucleic Acid Heteroduplexes; D009705:Nucleosides; D009841:Oligonucleotides; D016914:Ribonuclease H; D000212:Acyclovir