Study On The Mechanism Of Action Of Small Molecular Structural Analogues With FTO And Its Inhibitory Effect On K562 Cells

FTO gene is related to the formation of human obesity,and the overexpression of FTO gene may also lead to the occurrence of a variety of diseases and cancers.Therefore,the research on small molecule inhibitors targeting FTO protein has become one of the hot spots in the fields of chemistry,biology,medicine and so on.In this paper,we studied the interaction mechanism between five series of small molecule compounds and FTO protein,providing theoretical basis for screening small molecule inhibitors of FTO.In this paper,the interaction mechanism of 4 isoflavones、14 acrylonitrile derivatives、3 flavanones、12 isoquinoline derivatives and 10 oxazoles with FTO protein was studied by spectrophotometry and molecular docking method.Studies have shown that overexpression of FTO can increase the incidence of leukemia,and FTO may be a target protein of leukemia.Therefore,the inhibitory effects of 14 acrylonitrile derivatives、12 isoquinoline derivatives、10 oxazoles on the activity of leukemia K5 62 cells were studied by CCK8 method.The research content is mainly divided into the following parts:In chapter one,the research method of protein-small molecule interaction and the CCK8 method of detecting the inhibition of small molecule compounds on the activity of leukemia cells are summarized.In chapter two,we studied the interaction between four isoflavones and FTO protein by fluorescence spectroscopy,circular dichroism spectroscopy and molecular docking.The results showed that the four isoflavones bound to FTO protein through the static quenching mechanism,and the conformation of FTO protein changed.Biochanin A has the strongest binding ability to FTO protein,and steric hindrance can affect the binding process of small molecules to FTO protein.In the binding process of Biochanin A and FTO,the hydrophobic force is the main force,while the main forces between Sophoricoside、Daidzin、Genistin and FTO are hydrogen bond and van der Waals force.In chapter three,the interaction between acrylonitrile derivatives and FTO protein was studied by spectrometric method and molecular butting technique,and the inhibitory effect of acrylonitrile derivatives on K562 cells was detected by CCK8 assay.By comparing the binding constants of acrylonitrile derivatives and FTO protein,it was found that the order of the binding ability of derivatives and FTO protein was:1e>1h>1m>1d>1g>1c>1f>1k>1b>1i>11>1j>1n>1a.By comparing the molecular structure,it was found that compounds containing fluorine、chlorine、bromine、methoxy 1e、1h、1m had stronger binding ability to FTO protein.At the same time,it was found that 1e、1h、1m had a better inhibitory effect on the proliferation of K562 cells.In chapter four,the interaction between 7-hydroxy-flavanone、6-hydroxyflavanone and 6-methoxy flavanone with FTO protein was studied.The results showed that 6-hydroxy-flavanone had the strongest binding ability with FTO protein,which changed the conformation of FTO protein.The steric hindrance may affect the binding ability of small molecules with FTO protein.The hydrophobic force between 7hydroxyflavanone、6-hydroxyflavanone and FTO protein is the main force.Hydrogen bond and van der Waals force are the main forces between 6-methoxy flavanone and FTO protein.In Chapter five,we studied the interaction of 12 isoquinoline derivatives with FTO protein.The binding ability of isoquinoline derivatives to FTO was as follows:2k>2j>2i>2e>2f>2c>2a>2g>2b>2d>2h>21,among which 2k、2j、2i had better inhibitory effect on leukemia K562 cells.In chapter six,the interaction between 10 oxazole compounds and FTO protein was studied.The experimental results showed that the interaction mechanism between oxazole compounds and FTO protein was a static quenching mechanism,and the order of their ability to bind to FTO protein was:3j>3d>3b>3e>3g>3i>3f>3h>3c>3a.Cytotoxicity test results showed that compounds 3b、3d、3j had better inhibitory effects on the proliferation of K562 cells.