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Application of laminaria japonica/laminarin extracts to preparing EV71 resistant medicine

The technology of laminarin and extract is applied in the application field of preparing anti-EV71 drugs, and can solve the problems of EV71 infection without effective treatment drugs and vaccines, etc.

Inactive Publication Date: 2015-12-16
INST OF BASIC MEDICINE OF SAMS +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no effective treatment and vaccine for EV71 infection

Method used

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  • Application of laminaria japonica/laminarin extracts to preparing EV71 resistant medicine
  • Application of laminaria japonica/laminarin extracts to preparing EV71 resistant medicine
  • Application of laminaria japonica/laminarin extracts to preparing EV71 resistant medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Research on the anti-EV71 effect of laminaria (kelp) polysaccharide water extract

[0030] (1) Preparation of sea-tangle (kelp) polysaccharide water extract:

[0031] Take 200g of kelp (kelp), wash it with PBS, homogenize it with a tissue homogenizer, add 500ml of deionized water and boil for 15min; filter the solid tissue through gauze and centrifuge at 10000g for 10min, and the supernatant (435ml) is kelp (kelp) Sugar water extract. Using glucose as a standard, the polysaccharide content in the extract was determined by the anthrone sulfate method to be 2.68 mg / mL.

[0032] (2) Cell strains and virus strains:

[0033] HEL( figure 1 ) is an EV71-sensitive host cell, prepared and preserved by our laboratory; EV71 virus was isolated by our laboratory from fecal samples of children with hand, foot and mouth disease in Jinan Hospital of Infectious Diseases, and the full sequence has been determined and submitted to NCBI (GenBank: JQ074187.1). Virus lesions o...

Embodiment 2

[0039] Example 2: Research on the anti-EV71 effect of kelp (kelp) polysaccharides extracted from alcohol and precipitated matter

[0040] (1) Preparation of laminaria (kelp) polysaccharide water extraction and alcohol precipitation:

[0041] Take 200g of kelp (kelp), rinse it with PBS, homogenize it with a tissue homogenizer, add 500ml of deionized water and boil for 15min; filter out solid tissue with gauze and centrifuge at 10000g for 10min, add absolute ethanol to the supernatant to a final concentration of 85% After standing still at room temperature for 20 minutes, it was centrifuged at 10,000 g for 10 minutes to obtain 0.758 grams of precipitation, which was the water-extracted alcohol precipitation of kelp (kelp) polysaccharides. After the precipitation was dissolved in 50 ml of deionized water, glucose was used as a standard, and the concentration of polysaccharides determined by the sulfate anthrone method was 7.21 mg / mL.

[0042] (2) Cell strains and virus strains: ...

Embodiment 3

[0049] Example 3: Research on the anti-EV71 effect of kelp (kelp) polysaccharide DEAE cellulose column separation fraction

[0050] (1) Preparation of kelp (kelp) polysaccharide DEAE cellulose column separation fraction:

[0051] Take 200g of kelp (kelp), wash it with PBS, homogenize with a tissue homogenizer, add 500ml of deionized water and boil for 15min; filter the solid tissue through gauze and centrifuge at 10000g for 10min; pass through a DEAE cellulose column, wash with 3 times the column volume 0.3M NaCl After elution, use 0.4M NaCl to elute and collect the 0.4M NaCl eluted fraction, which is the laminaria (kelp) polysaccharide DEAE cellulose column separation fraction, desalt by ultrafiltration tube centrifugation and concentrate to 25ml, and measure the washing by anthrone sulfate method. The polysaccharide content of defractionated was 1.55mg / mL.

[0052] (2) Cell strains and virus strains:

[0053] HEL is an EV71 sensitive host cell, prepared and preserved by ou...

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Abstract

The invention discloses novel application of laminaria japonica / laminarin extracts. The novel application includes application of the laminaria japonica / laminarin extracts to preparing EV71 resistant medicine. A method for preparing the laminarin extracts can be one of a method (1), a method (2) and a method (3). The method (1) for preparing the extracts from laminaria japonica water includes washing laminaria japonica by the aid of PBS (poly butylenes succinate); boiling the laminaria japonica in double-distilled water; filtering the double-distilled water to obtain boiled juice; centrifuging the boiled juice to obtain supernatant so as to obtain the extracts. The method (2) for preparing aqueous extraction ethanol sediments from laminaria japonica includes washing the laminaria japonica by the aid of PBS; boiling the laminaria japonica in double-distilled water; filtering the double-distilled water to obtain boiled juice; adding anhydrous ethanol into the boiled juice until the concentration of the ethanol reaches 75-95% to obtain mixed liquid; centrifuging the mixed liquid to obtain the aqueous extraction ethanol sediments so as to obtained the extracts. The method (3) for preparing separation components of laminaria japonica by the aid of DEAE (diethyl-aminoethanol) cellulose columns includes separating the aqueous extraction ethanol sediments from one another by the aid of the DEAE cellulose columns; eluting the aqueous extraction ethanol sediments by the aid of NaCl solution; collecting eluted components to obtain the extracts. The novel application has the advantages that experimental bases are provided for clinically treating EV71 infectious diseases by the aid of the laminaria japonica extracts, and the application has a certain guidance significance on developing the EV71 resistant medicine and has important reference value.

Description

technical field [0001] The invention relates to the application of kelp / kelp polysaccharide extract in the preparation of anti-EV71 medicine. Background technique [0002] Kelp, also known as river cabbage and kelp, belongs to the genus Laminaria kelp of the Phaeophyta phytozoa, order Laminaria, brown algae, generally 2 to 4 meters long, and up to 7 meters long. It can be divided into three parts: the holder, the handle and the blade. The fork-shaped branch of the fixator is used to attach the rocks of the seabed. Stem short, cylindrical. The leaves are long and narrow, band-shaped. It grows in the sea with low water temperature, and is distributed along the coasts of northern China, North Korea, Japan, and the Pacific Ocean of the Soviet Union. It is known as "longevity vegetable", "vegetable of the sea", and "champion of iodine". Kelp is a kind of large brown algae with large individual, fast growth and high economic value. It is cultivated in large quantities in the n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/02A61K31/715A61P31/14
Inventor 李志会李鹏岳盈盈宋楠楠李冰清孟红秦立增
Owner INST OF BASIC MEDICINE OF SAMS
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