Human Immunodeficiency Virus (HIV) infects every year many people in the world. Unfortunately, the efficacy of the anti-virus used in clinic is very small. The project is focused on the three most used HIV reverse transcriptase inhibitors. The research has as its objectives the characterization of the molecular structure in water solution and to analyse the interactions with ATP and with RT-primer template. This study will give a new perspective in the knowledge of the three phosphorylation steps that are responsible of the low activity of the anti-HIV drugs used today. The process investigated could help for developing drugs with high anti-HIV activity and low toxicity. The expected impact will be high due to the millions of HIV infected people. The donor partner has a large experience in vibrational spectroscopy on biomolecules and biological material. Norway group has several IR and Laser-Raman spectrophotometers of great accuracy. The collaboration will help the scientific community in this important area of antivirus drug-design using theoretical quantum chemical methods.
Summary of project results
Human Immunodeficiency Virus (HIV) infects every year many people in the world, and unfortunately, the efficacy of the anti-virus used in clinic is very small. The project focused on the three most used today HIV reverse transcriptase inhibitors D4T, AZT and Nikavir, and in the three phosphorylation steps by the corresponding thymidine kinase enzymes. Thus, due to the present project these steps are being simulated through a simplified model that includes the antiviral molecule and the ATP molecule. Aims of the project have been: 1) The determination of the most stable conformer in the isolated state and in water solution of the anti-HIV nucleosis analogues AZT, D4T and Nikavir, as well as the natural nucleoside deoxythumidine (dT) and the adenosine 5’-triphosphate (ATP) molecule. 2) The accurate characterization of all the bands of the experimental Raman spectra in water solution of all the molecules under study, as well as the IR bands using the special microhydration technique developed by promoters. 3) The simulation of the crystal unit cell of all the molecules under study, and the further determination of their IR and Raman spectra. 4) The resolution of the first phosphorylation step mechanism of the anti-HIV nucleosides under study, as well as on dT, through a simplified model including the interaction with the optimum structure of ATP, firstly in the isolated state, and then in water solution with different number of water molecules surrounding the system. 5) The experimental interpretation of the interaction of ATP with the molecules under study in the first phosphorylation step through an accurate characterization of all the bands in the IR and FT-Raman spectra. 6) The determination of the second and third phosphorylation step mechanisms of the mono- and diphosphate forms of the anti-HIV nucleosides under study, as well as on dT, through a simplified model which includes the interaction with the optimum structure of ATP, firstly in the isolated state, and then in water solution. 7) Detailed analysis of the last step through the bonding of the triphosphorylated anti-HIV nucleosides analogues in the DNA viral through the reserves transcriptase enzyme (RT) in a simplified mode of the enzyme cavity with few residues. A number of scientific papers have been published, prepared and submitted and others are in preparation. Works continue after NILS funding ending. http://nofima.no/en/prosjeckt/nils-projekt http://nils-ucm-netau.net
Summary of bilateral results
The project activities were performed in close cooperation, with Spanish researchers from University Complutense of Madrid working at the Nofima As during more than a year. Complementarily to the project research activity, partners organized a workshop entitled “Raman and fluorescence spectroscopy: applications in life sciences”. This workshop helped partners to establish a better collaboration between Norwegian and Spanish research environments within this field of research. At the end of the project, partners were planning new cooperation activities and preparing applications to be submitted to other funding schemes, as well as working on several joint publications.