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Trametes robiniophia Murr. polysaccharide protein and preparation method and application thereof

A technology of polysaccharide protein and Huaier polysaccharide, which is applied in the field of Huaier polysaccharide protein and its preparation, can solve the problems of lack of research data, inability to clarify the principle of action, inability to understand the process of polysaccharide receptor action, etc., to achieve uniform molecular weight, The effect of clear structural features

Active Publication Date: 2014-10-29
QIDONG GAITIANLI MEDICINES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Similarly, in the research on the biological activity of Huaier polysaccharide protein, crude polysaccharides are mostly used as the research object, and there is a lack of in vivo and in vitro pharmacological research data obtained from the application of uniform polysaccharide components, resulting in the inability to deeply understand the relationship between Huaier polysaccharide protein and human immune cells or tumors. The action process of related polysaccharide receptors on the cell surface, so the mechanism of action cannot be elucidated in depth

Method used

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  • Trametes robiniophia Murr. polysaccharide protein and preparation method and application thereof
  • Trametes robiniophia Murr. polysaccharide protein and preparation method and application thereof
  • Trametes robiniophia Murr. polysaccharide protein and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Separation and purification method of Huaier polysaccharide protein (TP-1)

[0046] Accurately weigh 300g of Huaier fungus extract Huaierqing paste (provided by Qidong Gaitianli Pharmaceutical Co., Ltd.), add appropriate amount of distilled water to dilute it into a dilute solution with a concentration of about 8% (mass / volume), and then use The Sevage method was used to remove free protein, and the operation was repeated 5 times. The sugar fraction (TCP) and the crude total free protein fraction (TFP) of Huaier were collected respectively. (TCP) After concentrating under reduced pressure to remove residual chloroform and n-butanol, carry out fractional alcohol precipitation. First, slowly add absolute ethanol to TCP to make it into a sugar solution with an alcohol volume concentration of 40% (volume). After standing at 4°C for 24 hours, centrifuge at 3000 rpm for 10 minutes. The precipitation number is TCP-40. Press and dry for further separation and purifi...

Embodiment 2

[0050] Example 2: Research on the Chemical Structure of Huaier Polysaccharide Protein (TP-1)

[0051] This example studies the chemical structure of Huaier polysaccharide protein TP-1 prepared in Example 1.

[0052] 1. Method for studying the chemical structure of Huaier polysaccharide protein (TP-1)

[0053] 1. Determination of purity and molecular weight

[0054] UPLC-GPC-ELSD instrument configuration and chromatographic conditions: American Waters UPLC, TSK-3000GPC column, autosampler, Millipore ultrapure water ion exchanger to prepare high-purity water (0.45μm cellulose acetate membrane filtration); flow rate 0.3ml / min .

[0055] Preparation of the standard curve: Weigh an appropriate amount of dextran standard substance, add deionized water, and configure it into a reference solution with a concentration of 0.5 mg / ml, and then perform UPLC detection one by one. The results are shown in image 3 .

[0056] Preparation of sample solution: Weigh a certain amount of TP-1,...

Embodiment 3

[0097] Example 3 Huaierqing ointment and its chemical components in vivo immune activity research

[0098] 1. Sample source

[0099] Huaier Qing Ointment was provided by Qidong Gaitianli Pharmaceutical Co., Ltd. The crude polysaccharide TCP, the homogeneous polysaccharide TP-1, TP-2, TP-3, TP-4, and the low molecular weight components TO-1, TO-2, TO-3, TO-4 were prepared by Example 1.

[0100] 2. Promote the proliferation of mouse splenocytes

[0101] 1) To detect whether Huaier polysaccharide can stimulate the proliferation of mouse splenocytes

[0102] Method: Each component of Huaier Qing Gao (crude polysaccharide TCP; Huaier polysaccharide TP-1, TP-2, TP-3, TP-4; low molecular weight components TO-1, TO-2, TO-3, TO -4; Huaier crude total free protein fraction TFP) and negative control drug dextran (Dextran) were co-cultured with mouse splenocytes, after 42h, mixed with 3 H-TdR, after 48h, the cell harvester Filterate harvester was used to detect its proliferation index...

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Abstract

The present invention provides trametes robiniophila polysaccharide protein, a preparation method therefor, and an application thereof. Monosaccharide compositions of the trametes robiniophila polysaccharide protein of the present invention are fucose, arabinose, galactose, glucose, xylose, and mannose, a mass ratio thereof is 3.2:2.0:33.5:5.5:1.2:50.4, and the weight-average molecular weight of the polysaccharide protein is 7.0x105-5.0x106Da and preferably is 2.69x106Da. The polysaccharide protein of the present invention can be used to prepare medicine for curing tumour.

Description

technical field [0001] The invention relates to a Huaier polysaccharide protein, a preparation method and application thereof. Background technique [0002] Huaier is the fruiting body of the fungus Trametes robiniophila Murr. in the Polyporaceae family. It is an important folk medicinal fungus with a history of more than 1600 years in China. The Ainsworth fungal classification system assigns it to the phylum Fungi, Basidiomycotina, Phytomycetes, Polyporaceae, and Trametes. The fruiting body of Huai Er is medium to large, corky, sterile, and is born on the trunks of broad-leaved trees such as Sophora japonica and Black locust. It is distributed in Hebei, Shaanxi, Liaoning, Hunan, Guangxi, Fujian and other places. It is a kind of tree that can cause Heartwood rot fungi. The taste is bitter and pungent, and its nature is flat and non-toxic. It has the effects of "curing wind", "breaking blood" and "reinforcing strength". It is clinically used to treat various diseases. Due ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C07K1/36C07K1/30C07K1/18C07K1/16A61K38/16A61P35/00
CPCA61K38/00C07K14/37A61P35/00C07K14/375
Inventor 徐无为陆正鑫
Owner QIDONG GAITIANLI MEDICINES
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