The Synthesis And Identification Of The Efficient And Specific Inhibitors,4-[(6-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-droquinazoline)Methyl]-N-(4-sulfamoylphenethyl) Benzamide And Its Derivative, Based On Oncogene SHP-2(PTPN11)Tyrosine Phosphatase

【background】Malignant tumor is a common disease in clinic, whose incidence andmortality rate has been raised. Main reasons causing the death of cancerpatients are the metastasis and recurrence, so far no satisfying treatment. Inrecent years, more and more studies show tumor occurrence and metastasis areclosely related to mutation and overexpression of SHP-2. But the mechanismof tumor occurrence, development, evolution and metastasis are not yet clear.SHP-2is a Src containing2homeodomain protein tyrosine phosphatase,which belongs to the protein tyrosine phosphatase family, has very importantrole for in the regulation of signal transduction and mediate various biologicalprocesses. Tyrosine phosphorylation is controlled by protein tyrosine kinaseand protein tyrosine phosphatase, regulating a variety of important cellularprocesses. Studying in the early stage of PTPs, it focuses on antagonistic effectof PTKs, in order to find the more important tumor suppressor gene. Amongthem, the discovery of human chromosome10deletion phosphatase and tensinhomolog (phosphatase and tensin homologue deleted on chromosome ten,PTEN, MMAC1, TEP-1)are the important sense of tumor research. With thedepth study, a variety of PTPs including SHP-2superfamily member as apositive regulator of signal transduction are found, who play a vital role in theproliferation and differentiation of normal cells and tumor cell proliferation, adhesion, transfer process. Studying of the gene structure and function ofSHP-2are continuously deepening. Cell survival and apoptosis maintainnormal growth and stability of the body’s internal environment. Cell growthfactor mediating signal transduction pathway plays an important role in thisinteraction process. Many biological processes including cell growth,proliferation and apoptosis are regulated by tyrosine phosphorylation. SHP-2as a widely expressed cytosolic proteins involves in signal transductionpathways, which mediated by cellur and growth factor, such as Ras-Raf-MAPK,JAK-STAT, PI3K/Akt pathway. In most cases, although SHP-2is a tyrosinephosphatase, but it works positive regulatory in the transduction of receptorPTK. In recent years, the mutation of SHP-2in solid tumors, leukemia andNoonan syndrome synthesis have been found in early stage and it is defined asthe oncogene tyrosine phosphatase. SHP-2has become the target of cancer andseeking effective drugs to inhibit the SHP-2mutation has become an importantdirection of tumor therapy. The highly specific small molecule inhibitors ofSHP-2have the very big development space. After a long time study, ourlaboratory screened9compounds of three kinds of structure may be combinedwith the molecular structure of SHP-2, or can inhibit the abnormal increase ofphosphatase activity. Compounds in which three kinds of representative named#162,#216,#220compounds. However, for such compounds could specificallyinhibits tumor cells whose SHP-2activity was significantly enhanced havingnot been reported in the literature. This experiment attempts to synthesis ofthese compounds and to detect its effect on tumor cells of SHP-2mutations,and provide reference for development of such drugs.【Methods】In order to study the effect of tumor cells with high SHP-2expression by target compounds, this paper synthesize the target compound#216:4-[(6-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-droquinazoline)methyl]-N-(4-sulfamoylphenethyl)benzamide and its derivative:4-[(6,8-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-droquinazoline)methyl]-N-(4-sulfamoylphenethyl)benzamide(Figure1). Based on the inverse synthetic method, the target product isdecomposed into three parts of brominated-2-amino benzoic acid, theisothiocyanate methyl benzoate and4-aminoethyl benzene sulfonic acidamide(Figure2). using anthranilic acid as raw material through brominationreaction synthesize brominated-2-amino benzoic acid; using p-aminobenzoicacid as raw material through esterification, substitution and Gabriel reactionsynthesize isothiocyanate methyl benzoate; using phenylethylamine as rawmaterials through acetylation, chlorosulfonation, ammoniation, hydrolysisreaction synthesize4-aminoethyl benzene sulfonic acid amide. Then usingbromo-2-amino benzoic acid, isothiocyanate methyl benzoate and4-ammon-iaethyl benzene sulfonic acid amide through cyclization, hydrolysis,condensation reaction synthesize the target compound.(Figure3).Then using the two target compounds to treat abnormal tumor cells ofhigh SHP-2phosphatase activity identify its high specific inhibition for SHP-2tyrosine phosphatase and discuss its possible mechanism.【Results】(1)The successful synthesis of#216compound:4-[(6-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-droquinazoline)methyl]-N-(4-sulfamoylphenethyl)benzamide and its derivative:4-[(6,8-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-droquinazoline)methyl]-N-(4-sulfamoylphenethyl)benzamide. Their structureswere confirmed by1H-NMR and ESI-MS.(2)Improve the synthetic route, improve yield and reduce cost.(3)Identify inhibition of proliferation of tumor cells with high activity of SHP-2by#216compound and its derivative treating and prove that#216compound its derivative can effectively inhibit proliferation of tumor cellswith high activity of SHP-2, and it show the time and concentrationdependence.【Conclusion】(1)The successful synthesis of#216compound:4-[(6-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-droquinazoline)methyl]-N-(4-sulfamoylphenethyl)benzamide and its derivative:4-[(6,8-bromo-4-oxo-2-thioxo-1,2,3,4-tetrahy-dro-quinazoline)methyl]-N-(4-sulfamoylphenethyl)benzamide. The target com-pounds were purified by column chromatography and confirmed theirstructures by melting point determination,1H-NMR and ESI-MS.(2)The synthesis method of#216compound and its derivatives is simple,many methods are used for the first time, and it is fit for the synthesis oflaboratory because low toxicity, high yield. The synthesis methods were notreported in the literatures, providing the basis and reference for the synthesisof these compounds.(3)Through the experimental verificates the#216compound and itsderivative have efficient, specific inhibition on SHP-2tyrosine phosphataseand inhibitit proliferation of tumor cells with high activity of SHP-2.