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Isatis root polysaccharides, and preparation method and use thereof

A technology of isatidis polysaccharide and Radix isatidis, which is applied in the field of medicine, can solve the problems of great differences in physical and chemical properties, no mention of the preparation process and physical and chemical properties of the total polysaccharide, and no reported biological activity, etc.

Active Publication Date: 2013-09-11
BEIJING ZHONGAN ADJUVANT BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] It can be seen from the above documents: (1) Radix isatidis water extract and total polysaccharides have vaccine adjuvant activity, but the preparation process and physical and chemical properties of the total polysaccharides are not mentioned, and the total polysaccharides obtained by different preparation methods have the same physical and chemical properties. may vary greatly
(2) It is reported in the literature that two purified polysaccharides were isolated from Radix Radix, with molecular weights of 11700 and 2.24×10 5 , but no biological activity has been reported

Method used

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  • Isatis root polysaccharides, and preparation method and use thereof
  • Isatis root polysaccharides, and preparation method and use thereof
  • Isatis root polysaccharides, and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Embodiment 1: the preparation of polysaccharide S1, S3 and S4

[0106] Chinese herbal medicine Radix isatidis 1kg, crushed, soaked in 10L of 75% ethanol at room temperature for 24 hours, filtered, centrifuged (3000r / min×10min), the residue was leached again under the same conditions, the filtrate was combined, and the extract was recovered under reduced pressure at 40°C-45°C. After drying the Radix isatidis residue extracted with 75% ethanol at 50°C, add 15L of distilled water and extract at 50°C for 5 hours, stirring from time to time; Radix isatidis residue was extracted for the second time under the same conditions. The water extracts extracted twice were combined, concentrated under reduced pressure at 50° C. to 55° C. to 1000 ml, and then 3 times the volume (3000 ml) of 95% ethanol was added for alcohol precipitation for 72 hours. Centrifuge the alcohol precipitation solution (3000r / min×10min), add 1000ml of water to the precipitated part, stir and dissolve, centr...

Embodiment 2

[0110] Example 2: Physicochemical properties of polysaccharides S1, S3 and S4

[0111] 1. Molecular weight determination of polysaccharides S1, S3 and S4 (HPLC method):

[0112] Gel chromatography column: TSK-4000

[0113] Mobile phase: 0.1MNa2SO4

[0114] Flow rate: 0.6ml / min

[0115] Detector: Differential

[0116] S1, S3 and S4 molecular weight measurement results see Figure 5 , Image 6 with Figure 7 .

[0117] The results of molecular weight determination showed that the peak molecular weight of polysaccharide S1 was 3600Da, the peak molecular weight of S3 was 66400Da, and the molecular weight of S4 peak height was 6460Da.

[0118] 2. Determination of monosaccharide composition of polysaccharides S1, S3 and S4 (gas chromatography)

[0119] S1, S3 and S4 are completely acid hydrolyzed, reduced and acetylated to produce sugar alcohol acetate derivatives, which are analyzed by gas chromatography. The gas chromatographic results of monosaccharide standards and S1, S...

Embodiment 3

[0133] Example 3: Adjuvant activity of polysaccharides S1, S3 and S4 to influenza vaccines

[0134] The polysaccharides S1, S3 and S4 prepared in Example 1 were used as vaccine adjuvants (0.1 mg / mouse in all doses), respectively mixed with H1N1 influenza vaccine (H1N1 influenza virus lysate, 30 μg / ml, purchased from Beijing Kexing Biological Products Co., Ltd. company) mixed with intramuscular injection to immunize mice, and the antibody titer was determined by ELISA method after 2 weeks of immunization. The experiment was divided into normal saline group, H1N1 vaccine group, H1N1+S1 group, H1N1+S3 group and H1N1+S4 group. The results show that the H1N1 virus lysate is an immune antigen, and polysaccharides S1, S3 and S4 can produce higher titer specific antibodies as adjuvants for the first immunization (see Figure 19 ). Increased antibody subtypes IgG, IgM, IgG2a, and IgG2b (see Figure 20 ).

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Abstract

The invention belongs to the technical field of medicine, relates to polysaccharide components extracted from isatis roots, and in particular relates to polysaccharide components S1, S3 and S4 extracted from isatis roots. The invention also relates to a preparation method of the polysaccharide components S1, S3 and S4, and use thereof as a vaccine adjuvant or an immunomodulator or for preparing a vaccine composition. The results of activity experiments indicate that the separated polysaccharide components S1, S3 and S4 provided by the invention have vaccine adjuvant activity and immunoregulation effec, thereby providing a new option for immunologic adjuvants.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to polysaccharide components extracted from Radix Radix, in particular to polysaccharide components S1, S3 and S4 extracted from Radix Radix, the invention also relates to a preparation method of the polysaccharide components S1, S3 and S4, and their use It is used as an immune adjuvant, an immune modulator or in the preparation of a vaccine composition. Background technique [0002] Isatis indigotica is the dried root of the cruciferous plant Isatis indigotica. It has the effects of clearing heat and detoxifying, cooling blood and relieving throat. Modern pharmacological studies have shown that Banlangen has the functions of improving immune function and anti-tumor. The main chemical components in Radix Radix are flavonoids, lignin, alkaloids and polysaccharides. In recent years, the following literatures have reported the effects of Radix isatidis water extract and total polysacc...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K39/39A61K39/395A61K31/715A61P37/02
Inventor 单俊杰王玉霞朱婷董娜李倩武军华姜威赵修南贾培媛
Owner BEIJING ZHONGAN ADJUVANT BIOTECH
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