Chris Savich and Mark Santoyo
LSSU Chemistry and Biology Students
Chris Savich and Mark Santoyo, were awarded a grant from the Fund for LSSU to purchase an EGFR enzyme system, ADP-Glo Kinase Assay Kit, and Staurosporine, so that they could conduct an interdisciplinary research project. The project consisted of Chris Savich synthesizing eight different 3,4-Dihydroquinazolines compounds through novel organic synthetic techniques, and then providing the compounds to Mark Santoyo to test for anti-cancer properties through the inhibition of the EGFR pathway.
Figure 1. Above is a picture of Chris Savich and Mark Santoyo
During the spring and early summer of 2017, Christopher Savich under the advisorship of Dr. R. Adam Mosey employed novel organic synthetic techniques to make a total of eight 3,4-Dihydroquinazolines. 3,4-Dihydroquinazolines are a class of nitrogen containing aromatic compounds that are of particular interest for their pharmaceutical properties, such as anti-fungal, anti-bacterial, and in the case of this study, their possibility for anti-cancer properties. Eight different 3,4-Dihydroquinazolines were made through a novel four step one-pot synthesis. Each of the eight 3,4-Dihydroquinazolines had different functional groups attached to the left side of the molecule so as to test which of the functional groups would produce the greatest effect.
During the summer of 2017, Mark Santoyo under the advisorship of Dr. Martha Hutchens began the second half of the project. Mark’s objective was to test whether any of the eight 3,4-Dihydroquinazoline compounds that Chris synthesized could inhibit the activity of an enzyme called EGFR (epidermal growth factor receptor) kinase. Excessive or uncontrolled activation of this enzyme contributes to the development of certain cancers. EGFR converts ATP to ADP, so if the enzyme was inhibited, less ADP would be produced. First Mark used the ADP-Glo Assay Kit to construct a standard curve relating luminescence intensity to ADP concentration. Next, he used the ADP-Glo kit together with the EGFR Enzyme System to optimize the concentration of EGFR kinase and its substrate for maximum ADP production. Mark then conducted the reaction but introduced a known kinase inhibitor Staurosporine, which was used as a positive control, providing data on how much inhibition would be needed from the 3,4-Dihydroquinazoline compounds to be considered significant. (Figure 2)
Figure 2. Above is a picture of Mark micropipetting a master mix into a 96 well plate.
Once the reference data was established and compounds were ready and purified (Figure 3), the reactions were repeated but instead of using Staurosporine, the 3,4-Dihydroquinazoline compounds were used. ADP production was measured by change in luminescence, read via a microplate reader. The result of this experiment were that the compounds exhibited no significant inhibition of ADP production. However, provided key information that can aid further research in the search for anti-cancer compounds by eliminating compounds with functional groups of the variety tested from the pool of possible functional groups.
Figure 3. Above is a picture of the compounds 51,53, 57 after purification.
In conclusion, Chris and Mark were given the ability to, respectively, utilize and practice organic synthetic chemistry in an applied setting and molecular biology techniques in a manner that came together to further our interdisciplinary understanding of signal transduction pathways, and their importance in current cancer research. We would like to thank the Fund of LSSU Grant for this great opportunity given to us, allowing us to further not only our education but also the understanding of cancer and the possible treatment of it.
The 250 MHz GPR mapped lakeward-sloping sediment units, interpreted as former foreshore deposits, topped by horizontal units, the product of wind processes. Data reveal changes to this basic architecture due to onlap, truncation, and erosion of surfaces. This is caused by influences of different water levels on sedimentation throughout the decades. A beach chronology based on aerial photos, derived from GIS technologies, show that patterns of onlap mapped in GPR coincide with lake-level rise.