Quinoline-4-carboxamide skeleton derivative and application thereof
A compound and unsubstituted technology, applied in the field of chemical medicine, can solve the problems such as little understanding of tumor occurrence, and achieve good stimulating effect, good anti-cancer and good selectivity
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Embodiment 1
[0354] Enzyme activity agonistic test of the compound of embodiment 1
[0355] The purpose of this experiment is to detect the agonistic activity of compounds on sirtuin in vitro. In this experiment, the fluorescence intensity detection method was used to test the activity of SIRT6 in vitro. MDL-800 is the reference molecule (or called positive control). The sirtuin agonist activity of the test compound was measured by EC 50 (half agonistic concentration or the activation rate of the test compound to the sirtuin SIRT6 activity at a concentration of 20 μM). EC 50 Values can be calculated from the activation rate of sirtuin activity by the test compound at a range of different concentrations.
[0356] 1.1 Experimental materials
[0357] Human SIRT6 enzyme (constructed in E. coli M15[pREP4] with vector pQE80L.1), the buffer is improved Buffer, 100% DMSO (dimethyl sulfoxide), NAD (nicotinamide adenine dinucleotide) , acetylated peptide substrate, trypsin and microplate rea...
Embodiment 2
[0390] Embodiment 2 compound 12q in vitro enzyme level for the selectivity test experiment of the same family
[0391] The purpose of this experiment is to detect the agonistic activity of compound 12q on sirtuins SIRT1, SIRT2 and SIRT3 and other HDAC families in vitro. The sirtuin agonist activity of the test compound was measured by EC 50 (half median agonist concentration). EC 50 Values can be calculated from the agonism of the sirtuin activity by the test compound at a range of different concentrations.
[0392] 2.1 Experimental materials
[0393] Human SIRT1 enzyme (BPS Biotechnology Co., Ltd., product number 50012), human SIRT2 enzyme (BPS Biotechnology Co., Ltd., product number 50013), human SIRT3 enzyme (Cayman Company, product number 10011194), the buffer is improved Tris-Buffer (Tris-labeled buffer), 100% DMSO (dimethyl sulfoxide), NAD (nicotinamide adenine dinucleotide), acetylated polypeptide substrate, trypsin and microplate reader (Synergy MX), above Raw m...
Embodiment 3
[0405] Example 3 The in vitro activity of compound 12q on human pancreatic cancer cell line PANC-1 and BxPC-3 cells
[0406] The purpose of this experiment is to detect the anti-proliferation level of compound 12q on cells in vitro. In this experiment, human pancreatic cancer PANC-1 cells and BxPC-3 cells cultured in the laboratory were used. After being treated with drugs for a certain period of time, MTT (tetramethylazozolium salt) colorimetry, cell clone formation, and EDU incorporation tests were used. , cell migration assay, cell cycle and apoptosis assay and Western Blot method detection.
[0407] 3.1 Experimental materials
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[0410] 3.2 Experimental method
[0411] 3.2.1 Cell lines and culture
[0412] Human pancreatic cancer cell lines BxPC-3, PANC-1, AsPC-1 and MIAPaCa-2 were purchased from American TypeCulture Collection (ATCC, Rockville, MD, USA). It was cultured in DMEM supplemented with 10% FBS (Gibco, Eggenstein, Germany), 100 units / mL ...
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