Wisconsin Alumni Research Foundation

Drug Discovery & Development
Drug Discovery Development
Methods for Synthesizing Natural and “Unnatural” UDP- and TDP-Nucleotides
WARF: P04021US

Inventors: Jon Thorson, Dimitar Nikilov

The Wisconsin Alumni Research Foundation (WARF) is seeking commercial partners interested in developing methods of modifying nucleotidylyltransferases to vary their substrate specificity.
Many current therapeutic drugs originated as glycosylated bacterial secondary metabolites that consist of a central core structure (aglycon) and various sugar (glycosyl) attachments. Nucleotidylyltransferases activate sugar phosphates as nucleotide diphosphosugar (NDP-sugar) donors during metabolite biosynthesis.

Because the sugar moieties define the biological activities of many of these metabolites, altering these carbohydrate ligands can produce valuable new pharmaceuticals. However, there is a lack of efficient methods to synthetically alter the carbohydrate ligands.
The Invention
UW-Madison researchers have developed methods of modifying nucleotidylyltransferases to vary their substrate specificity in a directed manner. The researchers discovered a nucleotidylyltransferase from Salmonella enterica, called LT2 alpha-D-glucopyranosyl phosphate thymidylyltransferase (Ep), that displays unexpected promiscuity toward both its nucleotide triphosphate (NTP) and sugar phosphate substrates. Ep can convert a wide variety of phosphates to their corresponding dTDP- and UDP-nucleotide sugars.

The researchers used the 3-D structure and other molecular details of Ep substrate recognition to design nucleotidylyltransferase mutants with varying substrate specificity. The nucleotidylyltransferases are preferably mutated at one or more amino acids in their active sites, divalent cation binding sites and/or auxiliary sites.
  • Producing large numbers of known and novel bioactive metabolites in vitro for use as potential therapeutics for cancer or infectious diseases
  • May broadly impact efforts to understand and exploit the biosynthesis of glycosylated, bioactive natural products
Key Benefits
  • Can produce an enormous number of UDP- and dTDP-sugar substrates, including sugars that were previously difficult or impossible to produce in vitro and sugars that have never been produced before
  • Provides a rapid method of converting sugar phosphates to nucleotide diphosphosugars
  • Offers an effective method for constructing libraries of the desired UDP- and dTDP-nucleotide diphosphosugars for in vitro glycorandomization
Additional Information
Related Intellectual Property
For current licensing status, please contact Rafael Diaz at [javascript protected email address] or 608-960-9847