Antiviral and immunosuppressive nucleosides phosphorylation

Human adenosine kinase enzyme is a purine salvage enzyme that has a broad substrate specificity and provides the phosphorylation of purine/pyrimidine nucleosides and pyrimidine ribonucleosides. Adenosine kinase is critical for phosphorylation of adenosine analogs and provides first steps of activation of highly effective anti-viral and immunosuppressive prodrugs, such as mizoribine, tubercidin and ribavirine

Principle

PRECICE® ADK Phosphorylation Assay Kit is based on competitive inhibition of suboptimal substrate inosine (IR) phosphorylation. In the absence of nucleoside competitor, adenosine kinase phosphorylates inosine resulting in the formation of IMP. IMP is further oxidized by IMPDH to XMP and NADH2 continuously monitored at 340nm. In the presence of nucleoside competitor, the phosphorylation of inosine, poor ADK substrate, is inhibited detected as a decrease in NADH2 formation.
PRECICE® ADK Phosphorylation Assay Kit is a first tool for rapid evaluation of substrates properties of novel nucleoside analogues for human adenosine kinase.

ADK phosphorylation reaction schema

PRECICE® ADK Phosphorylation Assay Kit

#K0507-02
#REF SIZE PRICE
#K0507-02 1 plate (96 assays) 420.00 € Inquiry

Updated on October 3rd, 2024.

Kit is provided in stable lyophilized form and shipped without dry ice

Download ADK Phosphorylation Assay protocol (User manual)

Validated with antiviral nucleoside analogues

Such as mizoribine, tubercidine, ribavirin.

Nucleoside Analogues Mizoribine Tubercidine Ribavirin
IC50
(Competitive Inhibition)
45µM 90µM 550µM

Simple

Homogenous and continuous (add-and-measure)

High-Throughput Analysis

The assay allows the test of 12 analogues at 7 different concentrations at the same time (or 6 analogues in duplicata)

Readout is peformed with plate reader fitted with 340nm filter

Scientific Works citing NOVOCIB Adenosine Kinase and PRECICE® ADK Assay kits:
  1. M. Orlicka-Płocka, A. Fedoruk-Wyszomirska, D. Gurda-Wozna, P. Pawelczak , P. Krawczyk, M. Giel-Pietraszuk, G. Framski, T. Ostrowski, E. Wyszko Implications of Oxidative Stress in Glioblastoma Multiforme Following Treatment with Purine Derivatives. Antioxidants 2021, 10, 950.
  2. L.M. Johnson, O.J. Smith, D.A. Hahn, C.F. Baer. Short-term heritable variation overwhelms 200 generations of mutational variance for metabolic traits in Caenorhabditis elegans. Evolution. 2020 Nov;74(11):2451-2464.
  3. K. DANIELYAN, R. D. VARDANYAN, A. SIMONYAN, A. S. SAGYAN The sole role of PRPS-1 in the regenerative processes after experimental stroke Poster#: 459.07/A7 2017 Neuroscience Meeting Planner.
    Washington, DC: Society for Neuroscience, 2017.
  4. U. Nayar, J. Sadek, J. Reichel, D. Hernandez-Hopkins, G. Akar, P.J. Barelli, M.A. Sahai, H. Zhou, J. Totonchy, D. Jayabalan, R. Niesvizky, I. Guasparri, D. Hassane, Y. Liu, S. Sei, R.H. Shoemaker, J. D. Warren, O. Elemento, K.M. Kaye, E. Cesarman. Identification of a nucleoside analog active against adenosine kinase-expressing plasma cell malignancies. J Clin Invest. 2017 Jun 1;127(6):2066-2080.
  5. K.S. Toti, D. Osborne, A. Ciancetta, D. Boison, K.A. Jacobson. South (S)- and North (N)-Methanocarba-7-Deazaadenosine Analogues as Inhibitors of Human Adenosine Kinase Kiran J. Med. Chem. 2016, 59, 14, 6860-6877.
  6. M.K. Bjursell, H.J. Blom, J.A. Cayuela, M.L. Engvall, N. Lesko, S. Balasubramaniam, G. Brandberg, M. Halldin, M. Falkenberg, C. Jakobs, D. Smith, E. Struys, U. von Döbeln, C. M. Gustafsson, J. Lundeberg, A. Wedell. Adenosine Kinase Deficiency Disrupts the Methionine Cycle and Causes Hypermethioninemia, Encephalopathy, and Abnormal Liver Function. Am J Hum Genet. 2011 Oct 7; 89(4): 507-515.
  7. E. CESARMAN, U. NAYAR, J.D. WARREN, J. SADEK US20190225643A1 Novel nucleoside analogs and use thereof in therapeutic treatment. Patent application filed by Cornell University