A method for screening folic acid metabolism related drug
A technology for folic acid metabolism and drugs, which is applied in the field of screening drugs related to folic acid metabolism, which can solve the problems that compounds cannot be applied to in vivo experiments or clinical experiments, and cannot effectively evaluate the physiological response of drugs, and achieve strong efficacy, small side effects, and high screening success rate Effect
Active Publication Date: 2019-01-15
SHANGHAI CHEMPARTNER CO LTD
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Problems solved by technology
[0004]
The technical problem to be solved by the present invention is to overcome the inability to effectively evaluate the physiological response triggered by the drug in the cells by using the enzymatic reaction detection method to screen the drug targeting DHFR in the prior art, and the screened compound cannot be applied to downstream in vivo experiments or Provide a method for screening drugs related to folic acid metabolism due to defects such as clinical trials
Method used
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Embodiment 2
[0043] Verification of embodiment 2 detection method
[0044] According to the method in Example 1, use MV-4-11 (purchased from U.S. ATCC company), HL-60 (purchased from U.S. ATCC company) or HEK293 (purchased from U.S. ATCC company) to test Pemetrexed (Shanghai) Biotechnology Co., Ltd.] activity. Specific results such as Figures 5a-5c and shown in Table 1 below.
[0045] Table 1
[0046] cell
[0047] It can be seen that the activity of Pemetrexed can be tested by carrying out experiments according to this method.
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The invention discloses a method for screening a folic acid metabolism related drug. The method comprises the following steps: (1) mixing a cell line with stable physiological state and a compound tobe detected at different concentrations respectively, wherein the cell line has a serine hydroxymethyltransferase expression function; (2) adding stable isotope labeled serine and incubating until thereaction is complete; 3, obtaining the supernatant of the cell culture solution and detecting the content of glycine label by the stable isotope; (4) judging the activity of the compound to be detected by using the glycine content and the corresponding concentration of the compound to be detected. The screening success rate of the detection method is high, and the screened compounds have strongerefficacy and less side effects.
Description
technical field [0001] The invention belongs to the field of biological detection, in particular to a method for screening drugs related to folic acid metabolism. Background technique [0002] Dihydrofolate reductase (DHFR, EC1.5.1.3) is an oxidoreductase that utilizes NADPH to reduce dihydrofolate (dihydrofolate) in organisms to produce tetrahydrofolate (tetrahydrofolate). Tetrahydrofolate and its derivatives are carriers of one-carbon units in cells and are essential coenzymes involved in the synthesis of DNA and amino acids. DHFR is an important target in drug development, and research on DHFR inhibitors has long been an important direction in the development of anticancer drugs: when tumors grow rapidly, they need folic acid-mediated one-carbon unit metabolism to provide sufficient DNA and protein synthesis Raw materials; DHFR inhibitors specifically combine with DHFR to block its function, so that dihydrofolate cannot be converted into tetrahydrofolate, hinder folic ac...
Claims
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IPC IPC(8): C12Q1/02G01N30/02
CPCG01N30/02G01N33/5008
Inventor 魏照阳孟俊伟
Owner SHANGHAI CHEMPARTNER CO LTD
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