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Inhibitor Screening - Contract Research Services (In vitro and Whole Cell)
Purine and pyrimidine nucleotides are universal metabolites of all earth-living organisms. Buiding blocks of DNA, RNA and coenzymes, they are also key players of numerous cell metabolism processes: energy vehicles, precursors of lipids and sugars biosynthesis, source of methylation, mediators of cell signalling and neurotransmission. Nucleosides and nucleotides metabolism, and therefore purine metabolism enzymes are therefore an important area of research in cell biology and in health science.
For the Drug Discovery industry, purine metabolism represents a rich source of validated or high-potential targets in Oncology, Infectiology and Immunology. The impact of drugs on nucleotides metabolism is a major concern for these therapeutic fields.
On the basis of its expertise in nucleotide metabolism and of its enzyme portfolio, NOVOCIB offers Contract Research Services for the screening of potential inhibitors. Screenings can be performed in vitro with purified recombinant enzymes, or in Whole-Cell assays. These Contract Research Services are provided under a Service Agreement.
Purine and pyrimidine nucleotides are universal metabolites of all earth-living organisms. Buiding blocks of DNA, RNA and coenzymes, they are also key players of numerous cell metabolism processes: energy vehicles, precursors of lipids and sugars biosynthesis, source of methylation, mediators of cell signalling and neurotransmission. Nucleosides and nucleotides metabolism, and therefore purine metabolism enzymes are therefore an important area of research in cell biology and in health science.
For the Drug Discovery industry, purine metabolism represents a rich source of validated or high-potential targets in Oncology, Infectiology and Immunology. The impact of drugs on nucleotides metabolism is a major concern for these therapeutic fields.
On the basis of its expertise in nucleotide metabolism and of its enzyme portfolio, NOVOCIB offers Contract Research Services for the screening of potential inhibitors. Screenings can be performed in vitro with purified recombinant enzymes, or in Whole-Cell assays. These Contract Research Services are provided under a Service Agreement.
| Purine Metabolism Enzyme Inhibition | ||
| Screening | Enzymes used | Aims |
| IMPDH Inhibition | Inosine Monophosphate Dehydrogenase
• Human Type II, recombinant, expressed in E.coli • Bacterial (Staphylococcus aureus) recombinant, expressed in E.coli |
Screening of IMPDH inhibitors. Inhibition kinetics and IC50 measurement. |
| PNP Inhibition | Purine Nucleoside Phosphorylase
• Human recombinant, expressed in E.coli |
Screening of PNP inhibitors. Inhibition kinetics and IC50 measurement. |
| Nucleoside Kinase Inhibition | ||
| Screening | Enzymes used | Aims |
| AK Inhibition | Adenosine Kinase
• Human, recombinant, expressed in E.coli |
Screening of AK inhibitors. Inhibition kinetics and IC50 measurement. |
| dCK Inhibition | Deoxycytidine Kinase
• Human recombinant, expressed in E.coli |
Screening of dCK inhibitors. Inhibition kinetics and IC50 measurement. |
| Coupled Assay | ||
| Screening | Enzymes used | Aims |
| Nucleoside Kinase-IMPDH Coupled Assay | Adenosine Kinase or
cytosolic 5'-nucleotidase (cN-II) and Inosine Monophosphate Dehydrogenase • Human recombinant, expressed in E.coli (eventually S. aureus IMPDH, recombinant, expressed in E.coli) |
Rapid evaluation of monophosphate forms of nucleoside analogues as IMPDH inhibitors |
| Screening | Principle | Aims |
| IMPDH inhibition | • Cell culture and treatment (with MPA for positive control) • Quantification of intracellular GMP, GDP, GTP and IMP by HPLC |
Validation of IMPDH inhibition in cultured cells |
| RNR inhibition | • Cell culture and treatment (with hydroxyurea or Gemcitabine for positive control)
• Quantification of intracellular deoxynucleotides di- and triphosphate by HPLC |
Validation of RNR inhibition in cultured cells |
| Multi-targeted Antifolates | • Cell culture and treatment (with Methotrexate for positive control)
• Quantification of intracellular nucleotides by HPLC |
Validation of the inhibition of purine and pyrimidne nucleotides de novo biosynthesis in cultured cells |