The drug Neovasculgen® is a highly purified
supercoiled form of the plasmid pCMV-VEGF165 encoding endothelial vascular growth factor (VEGF) under the control of the promoter (DNA control region). The recombinant plasmid DNA consists of the following components: a fragment of the regulatory region (22 nucleotide pairs) that determines the transcription of the gene, the mini VEGF gene, in the expression of which the 165-amino acid isoform VEGF, the splicing signal, the polyadenylation signal and the SV40 transcription terminator are synthesized synthesis of the mature RNA gene and the auxiliary regions necessary for the efficient biosynthesis of plasmid DNA in the cells of the E. coli strain. When molecules of this plasmid penetrate into the cells of mammals, VEGF is produced, which stimulates endothelial cells,which leads to the growth of blood vessels (vascularization) in the area of administration. Endothelial cells are involved in such diverse processes as vasoconstriction and vasodilation, presentation of antigens, and also serve as very important elements of all blood vessels, such as capillaries, veins or arteries.
Thus, by stimulating endothelial cells, VEGF plays a central role in the process of angiogenesis. There are two different, but structurally similar, VEGF receptors located on the surface of vascular endothelial cells. These receptors, known as the VEGF1 type receptor (Flt-1) and the VEGF type 2 receptor (KDR / Flk-1) are receptor tyrosine kinases that, after binding to the VEGF ligand, undergo phosphorylation. Activation of these receptors leads to the inclusion of numerous intracellular post-receptor signaling cascades,
triggering angiogenesis. Penetration of the plasmid into cells is a spontaneous process, and plasmid DNA can not reproduce in mammalian cells. The molecular mass of the plasmid is -2817091.85 g / mol. The size is 4559 base pairs.
The peculiarity of the supercoiled circular DNA is the sensitivity to rupture of one of 9116 phosphodiester chemical bonds.A DNA molecule with a single broken bond, loses supercoiling, passing into a relaxed annular form (a related compound). In the experiment, Neovasculgen® stimulated the proliferation of human endothelial cells, comparable to a recombinant vascular endothelial growth factor, and Neovasculgen® significantly stimulated the growth of mouse blood vessels. The drug has been developed as a therapeutic tool to reduce the frequency of amputation and mortality in a population of patients with chronic limb ischemia, especially in patients who can not undergo standard revascularization for occlusive peripheral arterial disease.