Preparation method and application of indole-3-formaldehyde

A formaldehyde and indole technology, applied in the field of medicine, can solve the problems of difficult separation of reactants and products, non-unique products, harsh experimental conditions, etc., achieve huge development potential and application prospects, strong xanthine oxidase inhibitory activity, steps simple effect

Active Publication Date: 2018-10-09
HAIKOU EXPERIMENTAL STATION CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1945, the Blume group produced indole-3 carboxaldehyde by Reimer-Tiemann reaction, obtaining a yield of 52%, but the reaction selectivity was poor and the product was not unique (Blume R C, Lindwall H G.J.Org.Chem., 1945,10:255 )
In 1994, Macor etc. used acid chloride and Grignard reagent to synthesize 3-formylindole, but Grignard reagent is a strong nucleophile, which is easy to react with acid chloride, which also causes the low yield of this reaction method (45 %) (Macor J E, Blank D H, Fox C B, etal.J.Med.Chem.,1994,37:2509)
In 2010, Taylor et al. improved the Friedel-Crafts acylation method for the synthesis of 3-acylindole. Unlike the traditional Friedel-Crafts acylation reaction, the catalyst used in the improved reaction was DBN, so that the yield could reach 57%-100%, which solves the harsh conditions that require strict anhydrous in the traditional process, but the catalyst is expensive, the reactants and products are difficult to separate, the harsh experimental conditions and the lower yield limit its industrial application (Taylor JE, Matthew D J, Jonathan M.J.Org.Lett.,2010,12:5740201)

Method used

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  • Preparation method and application of indole-3-formaldehyde
  • Preparation method and application of indole-3-formaldehyde
  • Preparation method and application of indole-3-formaldehyde

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of indole-3-carbaldehyde

[0034] 1. Cultivation and fermentation of Vibrio New Caledonia

[0035]From the 2216E solid plate, use an inoculation loop to inoculate a single colony CGJ02-2 of Vibrio New Caledonia (deposited in the China Center for Type Culture Collection (CCTCC) on December 18, 2017, and the preservation number is: CCTCC M 2017802) to In 6 250 ml Erlenmeyer flasks each containing 100 ml of LB liquid medium containing 2% (g / ml) NaCl, cultivate overnight at 28° C. to 30° C., 160 rpm. The activated bacterial liquid was expanded and cultured with LB medium containing 2% NaCl at a ratio of 1:50, with a total volume of 16L, at 28°C to 30°C, and 120rpm for 10 to 12 days. The fermented liquid was centrifuged at 8000rpm for 10 minutes with a high-speed centrifuge, the bacterial cells were removed, and the supernatant was harvested.

[0036] 2. Extraction of fermentation broth

[0037] The culture supernatant was submerged in an equal volume of ethyl...

Embodiment 2

[0044] Determination of the Structure of Indole-3-Carboxaldehyde

[0045] Take part F6.2.2.1 obtained in Example 1, and use 2D-NMR (nuclear magnetic resonance technique) (BRUKER AVANCE 500MHz) for structural identification. in, figure 1 is the H-NMR spectrum, figure 2 for 13 C-NMR spectrum, image 3 is the COZY map, Figure 4 is the HMBC spectrum, Figure 5 It is the HSQC chart. Through structural analysis, the compound prepared by the present invention is indole-3-carboxaldehyde, and its chemical structural formula is:

[0046]

Embodiment 3

[0048] Determination of the Inhibitory Activity of Indole-3-Carboxaldehyde on Xantine Oxidase (XO)

[0049] Add 120μL of 0.02mol / L PBS (pH 7.4), 20μL of different concentrations of indole-3-carbaldehyde, 40μL of 1.5mmol / L xanthine, and 20μL of 0.5U / mL xanthine oxidase in a 96-well plate, and mix well , the absorbance was measured at a wavelength of 295nm with a full-wavelength microplate reader, and allopurinol was used as a positive control, and the average value was obtained by repeating 3 times. Enzyme activity experiment is set as experimental group, experimental blank group, negative control, negative blank control, positive control and positive blank control 6 groups.

[0050] Experimental blank group: 40μl xanthine + 20μl sample + 120μl PBS + 20μl PBS

[0051] Negative control: 40 μl xanthine + 20 μl PBS + 120 μl PBS + 20 μl enzyme

[0052] Negative blank control: 40μl xanthine + 20μl PBS + 120μl PBS + 20μl PBS

[0053] Positive control: 40 μl xanthine + 20 μl allopu...

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Abstract

The invention provides a method for preparing indole-3-formaldehyde. The method comprises the following steps: (1) inoculating vibrio neocaledonicus sp CGJ02-2 into an LB liquid culture medium and culturing; after the culturing is finished, centrifuging to remove thalli, so as to obtain fermented supernatant; (2) extracting the fermented supernatant by utilizing an organic solvent; decompressing and concentrating an organic phase to obtain crude extract; (3) separating the indole-3-formaldehyde from the obtained crude extract. According to the method provided by the invention, the indole-3-formaldehyde is prepared through a microorganism fermentation method; a lot of the indole-3-formaldehyde can be obtained only if microorganisms are cultured through a simple culture medium. The method provided by the invention is low in cost and is more environmentally friendly than a chemical synthesis manner. An in vitro enzyme activity experiment of the indole-3-formaldehyde shows that the indole-3-formaldehyde has high xanthine oxidase inhibition activity and has great development potential and application prospect in the aspects of treating hyperuricemia and gout.

Description

technical field [0001] The present invention relates to the technical field of medicine, in particular to a method for preparing indole-3-carboxaldehyde by microbial fermentation and the application of indole-3-carboxaldehyde in the preparation of medicines for treating hyperuricemia and gout . Background technique [0002] Uric acid is a product of purine metabolism in the human body. Due to the limited solubility of uric acid in body fluids, when purine metabolism in the body is disordered or excessive intake of purines leads to excessive purine content in the body, excessive uric acid is produced / or uric acid excretion is reduced, and the concentration of uric acid accumulated in body fluids When the saturated concentration is exceeded, crystals are easily precipitated in the tissue, causing hyperuricemia (hyperuricemia), and about 5% to 12% of patients with hyperuricemia eventually develop into gout (gout). Gout has become the second most common metabolic disease after ...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12P17/10C07D209/12A61P19/06C12R1/63
CPCA61P19/06C07D209/12C12P17/10
Inventor谭琳谭凯伊莎贝尔范梦凯孙香荣
OwnerHAIKOU EXPERIMENTAL STATION CHINESE ACAD OF TROPICAL AGRI SCI