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Medical application of granaticin type compound

A technology for compounds and uses, applied in the field of medical use of punicillin compounds, can solve the problems of different structures, low sequence similarity and the like

Inactive Publication Date: 2017-11-17
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, although IDO and TDO catalyze the same reaction, the two enzymes have very low sequence similarity and different structures (Badawy AbdullaA-B. et al, Bioscience Reports 2015;35:e00261)

Method used

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  • Medical application of granaticin type compound
  • Medical application of granaticin type compound
  • Medical application of granaticin type compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060]Example 1 Extraction and preparation of durianin compounds A-F of the present invention

[0061] Preparation of seed solution of N01W-352

[0062] The slant culture or spore liquid of strain N01W-352 was inoculated on the seed medium, and cultured at 28 °C and 200 rpm for 48 h to obtain the seed liquid;

[0063] The seed medium mentioned therein was prepared as follows: 20.0 grams of corn starch, 4.0 grams of glucose, 3.0 grams of peptone, 4.0 grams of beef extract, 2.0 grams of yeast extract powder, added water to 1000 mL; pH value 7.0, 121 ℃ Sterilize for 30 minutes;

[0064] Preparation of fermented broth of N01W-352

[0065] The above seed liquid was inoculated into the fermentation medium with an inoculation amount of 8% by volume, and fermented in a shake flask or a fermenter at a fermentation temperature of 27°C and a fermentation time of 120 h;

[0066] The fermentation medium mentioned therein was prepared according to the following method: 30.0 grams of gluc...

Embodiment 2

[0084] Example 2 Determination of the inhibitory activity of the duracin compounds of the present invention on IDO or TDO

[0085] Using genetic engineering technology, construct expression plasmids containing human IDO and human TDO genes, carry out recombinant expression in Escherichia coli, isolate and purify recombinant human IDO and TDO, and establish an activity detection system for IDO or TDO. Activity method of IDO or TDO Detection method: 96-well plate method is adopted, and the detection buffer contains 50 mM potassium phosphate buffer (pH 6.5) containing 40 mM vitamin C, 400 μg / ml hydrogen peroxide and 20 μM methylene blue. Add an appropriate amount of IDO or TDO to the detection buffer, make an enzyme mixed solution and add it to a 96-well plate, then add the substrate L-tryptophan, react at 37°C for 60min, add 30% (w / v) Trichloroacetic acid terminated the reaction. Incubate the 96-well plate at 65°C for 15 minutes to complete the conversion from formylkynurenine ...

Embodiment 3

[0088] Example 3 Inhibitory activity of durugin compounds of the present invention on IDO induced by gamma-interferon in Hela cells

[0089] Hela cells were cultured in a 96-well plate, the medium was high-sugar DMEM containing 50 U / ml penicillin, 50 U / ml streptomycin, 10% FBS, and the culture conditions were 37°C, 5% CO 2 . Stimulate Hela cells with γ-interferon at a concentration of 200IU, use IDO inhibitor 1-methyltryptophan (l-MT) as a positive drug, add different concentrations of positive drugs and compounds and 50 μM substrate L-tryptophan After culturing for 48 hours, remove the cell culture supernatant, add 30% (w / v) trichloroacetic acid, and incubate the reaction system at 65°C for 15 minutes to complete the transformation from formylkynurenine to kynurenine For conversion, centrifuge at 12,000rpm for 10min, take the supernatant and mix with an equal volume of 2% (w / v) acetic acid solution of p-dimethylaminobenzaldehyde to react, detect the OD value at 490nm, and ca...

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PUM

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Abstract

The invention belongs to the technical filed of medicinal chemistry and particularly relates to a medical application of a granaticin type compound. The compound is used for preventing or treating diseases, caused by IDO and / or TDO, with pathological characteristics related to tryptophan metabolism, wherein the diseases comprise but is not limited to tumor immune escape, or the virus infectious diseases, or the nervous system diseases, or the mental diseases or the cardiovascular diseases or the like.

Description

technical field [0001] The invention belongs to the technical field of medicinal chemistry, and in particular relates to the medical application of durianin compounds. Background technique [0002] Heme-containing proteins have diverse functions, including oxygen transport, storage, and one-electron transfer activation. Both indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) belong to the family of heme-containing proteins (Sedlmayr. et al, Frontiers in Immunology 2014;5:230) . TDO was first discovered around 1930. TDO is generally considered to be expressed only in the liver of mammals, but studies have confirmed that it exists in both eukaryotes and prokaryotes. IDO was discovered in 1967 and was isolated from the intestines of rabbits. Studies have confirmed that IDO also exists in many other eukaryotes (Ball HJ. et al, Front Immunol 2014;5:485). [0003] The active function of TDO and IDO is to catalyze the oxidation of tryptophan to formylkynureni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/365A61K31/352A61P35/00A61P35/02A61P31/14A61P31/20A61P31/22A61P31/16A61P31/18A61P25/28A61P25/14A61P25/16A61P21/00A61P9/00A61P25/22A61P25/24A61P25/00A61P25/20A61P25/18
CPCA61K31/352A61K31/365Y02A50/30
Inventor 郑智慧任晓张雪莲郑海洲路新华朱京童马瑛丁彦博穆云龙高健
Owner NCPC NEW DRUG RES & DEV
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