Evaluation of novel synthetic anillinoquinazolines as potential angiogenic modulators

Vahlas, Zoi

dc.contributor.author	Vahlas, Zoi	fr
dc.date.accessioned	2021-05-14T11:13:42Z
dc.date.available	2021-05-14T11:13:42Z
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/53433
dc.identifier.uri	http://dx.doi.org/10.26240/heal.ntua.21131
dc.rights	Default License
dc.subject	Angiogenesis	en
dc.subject	Vascular endothelial growth factor receptor (VEGFR)	en
dc.subject	Epidermal growth factor receptor (EGFR)	en
dc.subject	Anillinoquinazoline	en
dc.subject	Cancer	en
dc.title	Evaluation of novel synthetic anillinoquinazolines as potential angiogenic modulators	en
dc.contributor.department	Synthesis and Development of Industrial Processes	el
heal.type	bachelorThesis
heal.classification	Biochemistry	en
heal.classificationURI	http://skos.um.es/unesco6/2302
heal.language	en
heal.access	free
heal.recordProvider	ntua	el
heal.publicationDate	2015-09-17
heal.abstract	Angiogenesis is the sprouting of new blood vessels from pre-existing ones. It is an important process during the development of the embryo and the wound healing. The cells at the inner surface of the blood vessels, named endothelial cells (ECs), enter in division phase and create the new blood vessel. The angiogenesis plays also a key role in some diseases like in cancer. In order to grow, tumors need oxygen and nutriments to feed the cancer cells. Such feeding can be done only by blood vessel. Consequently, in this, as well as in other diseases involving angiogenesis, the division of the endothelial cells is upregulated so as to create new blood vessel to feed the cancer cells. This is the driving force behind thousands of studies which took place up to now with the aim to find a solution to downregulate this mechanism of anarchic, uncontrolled and incessant cell proliferation. The angiogenic process takes place through several steps: 1) Induction by growth factors such as the vascular endothelial growth factor (VEGF) and the epidermal growth factor (EGF). They are realized in response to different types of stress (hypoxia, cut and other). These growth factors bind to tyrosine kinase receptor like vascular endothelial growth factor receptor (VEGFR) or epidermal growth factor receptor (EGFR). 2) Stimulated Endothelial cells secrete protease (like plasminogen activator and collagenases) which locally degrades the basement membrane of the initial blood vessel and the extracellular matrix. 3) Endothelial cells proliferate and migrate through the formed gaps of the basal membrane in the direction of the angiogenic stimuli (known as chemotactic signal). Adhesion molecules (integrins), serve as grappling hooks in order to help the sprouting of new blood vessels. 4) The angiogenesis process is terminated with the formation of new basal membranes and the accumulation of pericyte cells (cells that inhibit the proliferation of endothelial cells) and other perivascular cells near the newly formed capillaries. From the perspective to inhibit the cell proliferation, some studies have been done and tyrosine kinase receptors inhibitors have been found to be very effective because they act on the region of tyrosine kinase of the growth factor receptor and inhibit it to be phosphorylated and to activate cell proliferation. The objective of this internship was to study 22 novel anillinoquinazoline compounds. They were chosen because they are tyrosine kinase receptor inhibitors and it has already been proved that they present an inhibiting effect on EGFR. It will be interesting to verify this inhibition for VEGFR, as well. The study was performed by using an in vitro model involving the fusion of adherent endothelial cells from fusing human umbilical vein endothelial cells (HUVECs) with cells of cancerous lung tissue (EAhy 926). The methods used in this internship were, first, the passage of cells. It is a method used to dilute the cells into a new Petri dish to prevent the confluence and cell death. Another method was the counting of cells, for this a hemocytometer and a microscope are used. Then, in order to assess the cell viability, the MTT assay is employed. It is a colorimetric assay which uses the NAD(P)H-dependent oxydoreductase enzymes produced in mitochondria of the cell to reduce the MTT. The result is in an insoluble product, the formazan which is break down by formazan. Only the viable cells can produce this enzyme so, by knowing the quantity of formazan, the number of living cells in suspension can be found. Lastly, in order to evaluate the results, a statistic analysis was used, the Mann-Whitney t-test analysis which informs about how close is the relationship between two price series. From the 22 compounds tested, it was found that four compounds present a promising impact to the inhibition of cell proliferation (72, 77, 168 and 80). Based on this result, it can also be expected that they present an inhibition action on angiogenesis. Moreover, those results have been compared with the results taken from Kostakis’s group unpublished data on the action of those compounds on cancer cell lines and it has been seen that the effects of the four compounds that are interesting for our research presented similar behaviors with the different cells lines even if the effect on the cancer cell lines was lower. In conclusion, it can be said that the four compounds (72, 77, 168 and 80) have an impact on cell proliferation probably by interacting with the VEGFR on angiogenesis and it seems that they have also an impact on cancer cell lines through the EGFR. The work performed in this internship is the first step of a research which aims at identifying quinazoline based compounds which inhibit angiogenesis. It is premature to make general conclusions from this work, because the behavior of cells in vitro is far from the realty and the complexity of the reactions that takes place in vivo. Also, because signaling pathways in which each compound act are very complex. Finally, the best concentration at which the compound must be administrated should be known. For those reasons, some perspectives are given in order to continue this work: • The IC50 of the compounds 72, 77, 168 and 80 must be determined. The IC50 is the concentration of a drug that gives half-maximal response. By this way, it will be clear which concentration must be used for each of these compounds to have a specific cell proliferation. • Then, it will be interesting to isolate the proteins of the cell cultured with the 4 compounds and to do a western blot in order to see the influence of these compounds on proteins involved in angiogenesis (EGFR, VEGFR…). • After, those compounds should be investigated in in vivo models like mice.	en
heal.advisorName	Topakas, E	en
heal.committeeMemberName	Topakas, E	en
heal.committeeMemberName	Boudouvis, A	en
heal.committeeMemberName	Kolisis, Fr	en
heal.committeeMemberName	Kekos, D	en
heal.academicPublisher	Εθνικό Μετσόβιο Πολυτεχνείο. Σχολή Χημικών Μηχανικών	el
heal.academicPublisherID	ntua
heal.numberOfPages	44 p.	en
heal.fullTextAvailability	true