Application of golden thread detoxication decoction as compound preparation in preparing IOD active medicaments
A technology of Huanglian Jiedu Decoction and compound preparation, applied in the field of traditional Chinese medicine, can solve problems such as lack of new IDO inhibitor drugs
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Embodiment 1
[0019] The preparation of recombinant IDO was carried out as described in the literature.
[0020] The IDO activity detection was carried out on a 96-well plate. First, 50 mM potassium phosphate buffer (pH 6.5), 40 mM vitamin C, 400 μg / ml catalase, 20 μM methylene blue and IDO enzyme were mixed. Then add the substrate L-tryptophan and the sample to be tested into the above mixed solution. The reaction was carried out at 37°C for 60 minutes, and the reaction was terminated by adding 30% (w / v) trichloroacetic acid. The 96-well plate was heated at 65°C for 15 minutes to complete the conversion from formylkynurenine to kynurenine, and then spun at 6000 g for 5 minutes. Take out 100 μl of supernatant from each well and transfer it to a new 96-well plate, add 2% (w / v) p-(dimethylamino)benzaldehyde in acetic acid solution, the yellow color produced by the reaction of kynurenine It can be observed at 490nm using a microplate reader.
Embodiment 2
[0021] Example 2. IDO inhibitory activity of Huanglian Jiedu Decoction
[0022] Coptis chinensis, Scutellaria baicalensis, Cortex Phellodendron and Gardenia were decocted twice according to the original ratio (3:2:2:3), adding 10 times and 8 times of water to decoct for 60 and 40 minutes respectively, combined the decoction liquid, concentrated to each One milliliter is equivalent to 1 gram of crude drug. The IDO inhibitory activity test of Huanglian Jiedu Decoction and IDO inhibitor 1-methyltryptophan (1-MT) was compared, and the results showed that the Huanglian Jiedu Decoction had obvious inhibitory efficiency on IDO.
[0023] Table 1. Inhibition efficiency of Huanglian Jiedu Decoction on IDO
[0024]
Embodiment 3
[0025] Example 3. IDO inhibitory activity of each single herb in Huanglian Jiedu Decoction
[0026] Each crude drug is soaked in ten times the volume of methanol for seven days, the soaking solution is concentrated into a paste, and dissolved in an appropriate amount of methanol, so that each milliliter of the solution is equivalent to 1 gram of crude drug. The IDO inhibitory activity of each crude drug was tested and compared with the IDO inhibitor 1-MT, and the results showed that each single drug in the Coptidis Jiedu Decoction showed inhibition of IDO activity, and Coptis chinensis had a significant inhibitory effect on IDO activity.
[0027] .
[0028] Table 2. Inhibition efficiency of each single drug in Huanglian Jiedu Decoction on IDO
[0029]
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