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A kind of pyridine-containing compound and its extraction method

An extraction method and compound technology are applied in the field of pyridine-containing compounds and their extraction, and can solve the problems such as no reports of compounds inhibiting COX-2.

Active Publication Date: 2021-01-15
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The compounds involved in the present invention have no reports on inhibiting COX-2 and anti-inflammation

Method used

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  • A kind of pyridine-containing compound and its extraction method
  • A kind of pyridine-containing compound and its extraction method
  • A kind of pyridine-containing compound and its extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Extraction, separation and structural identification of the compound of the present invention

[0024] Japanese Pipa A 50kg, crushed, extracted with 70% ethanol cold soaking for 3 times, each time for 48h, 6 times the volume, stirring continuously, filtering, recovering ethanol under reduced pressure, to obtain 4kg extract, suspending in acid water, adjusting pH 1–2 , extracted 4 times with equal volume of ethyl acetate to obtain 1 kg of non-alkaloid fraction. The pH of the remaining acidic aqueous layer was adjusted to neutral or slightly alkaline with alkali, and extracted 4 times with equal volume of ethyl acetate to obtain 300 g of alkaloid fraction. The alkaloid fraction (300g) was chromatographed on MCI gel CHP 20P column, methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20 , 90:10, 100:0) gradient elution, TLC detection and combining the same fractions to obtain Fractions A-E a total of 5 fractions. Fr.D (20 g) was separated by Seph...

Embodiment 2

[0029] Embodiment 2 compound inhibits COX-2 test method

[0030] In this experiment, Cayman’s COX Fluorescent Inhibitor Screening Assay Kit was used to detect the enzymatic activity of COX-2. Specific steps are as follows:

[0031] Compound gradient dilution: Dissolve the compound in DMSO into a 10mM stock solution, and then use DMSO to dilute it 1:3 into a 1000×compound serial concentration stock solution with 10 concentration gradients, and then transfer it to a 384PP plate adapted to the compound transfer instrument for later use.

[0032] Add 10 μl of enzyme diluent to each well, and add 10 μl of 1xassay buffer to blank control wells without adding enzyme.

[0033] The compound was transferred to 20 nl of 1000× gradient diluted compound and added to the reaction solution, and DMSO was added to the blank control and DMSO control wells. Incubate at room temperature for 10-15min.

[0034] Dilute the fluorescent substrate ADHP and coenzyme Heme with 1xassay buffer, the dilu...

Embodiment 3

[0038] The compound in Example 1 can be prepared into an injection solution after adding injection solvent according to the conventional method, finely filtering, filling and sterilizing.

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Abstract

The invention provides a pyridine-containing compound, relating to the field of medicines. The chemical structure of the compound is shown as in a formula I: The formula I is shown in the description.The compound can be taken as an inhibitor of cyclooxygenase 2 for treating inflammation. The invention further provides an extration method of the compound for preparing the pyridine-containing compound which has the effect of inhibiting cyclooxygenase 2 and has application value in the aspect of inflammation-related diseases.

Description

technical field [0001] The invention belongs to the field of medicines, and in particular relates to a pyridine-containing compound and an extraction method thereof. Background technique [0002] Since the listing of aspirin (ASA) in 1899, non-steroidal anti-inflammatory drugs (NSAIDs) have been used clinically for a hundred years. Until 1971, it was discovered for the first time that ASA exerts anti-inflammatory, antipyretic and analgesic effects mainly by inhibiting cyclooxygenase (COX) to synthesize prostaglandin (PG). Among them, cyclooxygenase is the rate-limiting enzyme that catalyzes the conversion of arachidonic acid into prostaglandins and related compounds. Currently, there are two common cyclooxygenases, COX-1 and COX-2, whose amino acid sequences share 60% homology. The difference is that COX-1 exists widely in various cells of the body, continuously expresses under physiological conditions and promotes the synthesis of prostaglandins to maintain some normal ph...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D213/30A61K31/4418A61P29/00
Inventor 程永现晏永明涂正超
Owner SHENZHEN UNIV
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