JPH0245501A - Chikusetsu ginseng polysaccharide and use thereof - Google Patents

Abstract

NEW MATERIAL:A compound having a mol.wt. of 20000-180000 and containing saccharides selected from arabinose, xylose, rhamnose, fucose, glucose, galactose and galacturonic acid as constituting components thereof. USE:A biophylaxis ability-accelerating agent. Low toxicity. PREPARATION:CHIKUSETSU ginseng (Panax Schinseng Nees var. japonicum Makino) belonging to the genus Panax in the family Arallaceae is ground and subsequently extracted with a water-soluble organic solvent such as methanol. The extraction residue is extracted with water and subsequently mixed with a water-soluble organic solvent such as ethanol to produce precipitates. The precipitates are finally subjected to a chromatography using an ion exchange resin or gel filtration agent.

Description

DETAILED DESCRIPTION OF THE INVENTION [Background of the Invention] Technical Field The present invention relates to a novel polysaccharide component isolated from ginseng and a biodefense enhancer containing the polysaccharide component as an active ingredient.

BACKGROUND OF THE INVENTION Bamboo knot ginseng is a perennial herb that inhabits China's Siyong and Yunnan provinces, and is used in China as a treatment for cough 1, blood, and rays.

Furthermore, regarding the components in bamboo knot ginseng, saponins such as Chiksetsusaponin L Ia, Ib, III and ■ have been isolated and their structures have been determined. However, there are almost no reports on polysaccharide components, and there is still room for research.

By the way, as humans grow older, their ability to resist various diseases generally decreases, making them more susceptible to various diseases. The cause is a decline in various internal organs and metabolic processes, as well as a decline in the function of phagocytes, which process foreign substances and waste.
It is thought that the body's defense ability declines.

As a result of repeated research on the components of Bamboo-bushi carrots, the present inventor discovered a new polysaccharide component in Bamboo-bushi carrots, and discovered that this polysaccharide component has biological defense ability, and was able to complete the present invention. It's arrived.

[Summary of the invention]

Summary The present invention aims to provide a novel polysaccharide component and a biodefense enhancer containing the polysaccharide component as an active ingredient, and is based on the unexpected discovery, even for those skilled in the art, that the polysaccharide component in bamboo-fried carrots has this effect. be.

Therefore, the polysaccharide derived from bamboo knot carrot according to the present invention is
It is characterized by having a molecular weight of about 20,000 to 180,000, and a constituent sugar selected from ``7-rabinose, xylose, rhamnose, fucose, glucose, galactose, and galacturonic acid.''

Furthermore, the biological defense ability enhancer according to the present invention has a molecular weight of about 2
0,000 to 180,000 and whose constituent sugars are selected from arabinose, xylose, rhamnose, fucose, glucose, galactose and galacturonic acid, as an active ingredient;
It is characterized by:

Effect The novel polysaccharide component of the bamboo-fried carrot according to the present invention has an excellent biological defense ability, and the biological defense ability enhancer according to the present invention containing this polysaccharide component as an active ingredient is effective against bacteria, viruses, etc. that invade the body. A substance that activates biological defense reactions such as phagocytosis of macrophages against foreign substances, increases the ability to eliminate foreign substances from the body, and thereby increases resistance to various diseases, and has low toxicity. It is. Therefore, the provision of a polysaccharide component and a biodefense enhancer containing the polysaccharide component as a nail-acting component according to the present invention will make a significant contribution to measures against various diseases related to bioresistance.

[Detailed description of the invention]

Bamboo-bush carrots Bamboo-bushi carrots as used in the present invention refer to Arallaceae (Arallaceae).
aceae), Panax Sehinseng Nees var belonging to the genus Panax
, japonlcum Makin.

(-P, japonlucus C, A, Mey,)
means.

The root is particularly preferred as the part from which polysaccharide components should be obtained;
Considering the acquisition efficiency, it is convenient to use dry ones. It is also possible to subject an appropriate part of a bamboo joint carrot to tissue culture by a conventional method and use the cultured product.

Polysaccharide component and method for obtaining the same The polysaccharide component derived from bamboo knot carrots as used in the present invention has a molecular weight of 20,000 to 180,000, and the constituent sugars are arabinose (Ara), xylose (Xyl), rhamnome (Rha), and fucose ( Fuc), glucose (G l
c) selected from galactose (Gal) and galacturonic acid (GalA). Therefore, this polysaccharide component essentially consists of only the above-mentioned sugars, and the constituent sugars may be only one of the above-mentioned sugars,
It may be a combination of multiple types.

Specific examples of such polysaccharide components are as follows.

In order to obtain the polysaccharide component as referred to in the present invention, any method capable of isolating the component from bamboobrush carrots may be used. For example, after crushing bamboo-fried carrots,
Extraction is performed by immersion in a water-soluble organic solvent, the residue is extracted with water, and a water-soluble organic solvent is added to obtain a precipitate. The desired product can then be obtained by subjecting the precipitate to chromatography using an ion exchange resin or a gel filter agent. The raw material carrot may be subjected to a defatting operation, and the desired product may also be obtained by subjecting the precipitate to dialysis.

For details regarding the isolation of this polysaccharide component, please refer to the Examples below.

Biodefense Enhancing Agent The biodefense enhancing agent of the present invention is composed of the polysaccharide component of Bamboo Bush Carrot itself or mixed with appropriate excipients, binders, and diluents for formulation, and can be used in powders, granules, tablets, It can be administered orally or parenterally in the form of capsules, syrups, injections, and the like. Further, other drugs may be mixed as necessary. The dosage may vary depending on age, body weight, and symptoms, but it is usually 0.1 to 0.1 b per day for adults per day as a polysaccharide component. It consists of Another preferred embodiment of the present invention is a unit dosage form in which the above-mentioned daily dose is divided into one or several doses.

In addition, the toxicity of the polysaccharide component in the present invention is, for example, in the case of PI3-2, PI3-3, PJN-1, and PJN-3 in the experimental examples described later, in the case of ICR male mice (5 mice).
No abnormalities were observed in general symptoms after intraperitoneal administration of 4 mg/kg to
．． 1084 (1981)) and PS-K (Krestin (Sankyo)), etc., are generally considered to have low toxicity, as they are well known to have low toxicity.

Enhancement of biological defense ability When foreign substances (bacteria, viruses, etc.) invade the living body, biological defense reactions such as phagocytosis of macrophages occur in response, and an ingenious mechanism is created to eliminate the foreign enemy. . However, when immunity or protective factors decrease, attacking factors (
foreign substances) become predominant, making you more susceptible to various diseases such as opportunistic infections.

In the present invention, the action of enhancing biological defense function refers to the action of activating the biological defense reactions shown above to eliminate foreign substances from the living body, and has secondary antiviral, antitumor, antiparasitic, and antiviral effects. It concerns the elimination of bacterial action and other foreign antigens.

Experimental example 1) Acquisition of polysaccharide components According to the process diagram in Figure 1, bamboo section carrots were mixed with methanol (M
The residue extracted with eOH) was dissolved in water and ethanol (EOH) was extracted.
DEAE-TOYOPEARL 65
P J N was applied to a column (40 mm x 700 mm) packed with 0M.

Obtain fractions of PJI, PI3, PI3, PI4,
The PJN and PI3 fractions were DEAE-T again.
PJN-1 (yield 0.28%), PJN-3 (yield 0.28%),
PJ2-2 (yield 0, 088%>, PJ2-3 (
(0,057%) was obtained (however, each yield is the weight % based on the initial bamboo-fried carrot (dry)).
.

The sugar content of these fractions was determined by the phenol-sulfuric acid method, the uronic acid content by the carbazole-sulfuric acid method, and the protein content by the Lovry method.

Further, in determining the constituent sugars, the ice was thawed with 1N sulfuric acid, reduced and acetylated by a conventional method, and then analyzed by gas chromatography (GLC).

In addition, in determining the binding mode of sugars, after methylation and acid hydrolysis, gas chromatography mass spectrometry (G C-M
Analyzed by S).

These results are shown below.

(i) PJN-1 Molecular weight: 23,000. Cα)D+49.5°(c-0
, 5, water) Elemental analysis: C39, 29%, H6, 36% N0.03% Protein analysis: Not detected (Lowry method) General FFI: 95.5% (standard material: Gal, phenol sulfuric acid method) Uronic acid Analysis: Undetected (carbazole sulfuric acid method) Acid hydrolysis: D-Gal Methylation analysis: Bonded to the 4-C position of Gal'H-NMR (in D20) δ: 4.45 (Ano 7-H, d, J -6,7Hz) 13C-NMR (in D20) δ: 105.1 (
C-1), 72.8 (C-2), 75.3 (C-
3), 78.3 (C-4), 74.2 (C-5)
, 61.6 (C-6).

Structure of PJN-1 β-1,4- is raftan.

n: positive integer N) PJN-3 Molecular weight: 40,000, [・'t'　+56. O.

(c-0,5, water) Elemental analysis: C37,47%, H6,22% NO,04% Protein analysis: Not detected (Lowry method) Total amount: 94.0% (phenol sulfuric acid method) (Mark material:
Gal, AraSGlc.

Ga1A) (26:2.4:0.5:1) (molar ratio) Uronic acid analysis, 3.5% (standard substance: Ga1A, carbazole sulfuric acid method) Reduction of carboxyl group (Taylor-Conrad reduction) PJN-3 to PJN-3R (reduced product) Acid hydrolysis = (IN H2SO4, 100°C, 4h) L-Ara D-Gal D-GlcPJN-31
,010,90,2 PJN-3 structure R = side chain 2 -Ga L-Glc, -Ara (f)-
(5”-1)-Ara (f) f: Furanose type (all others are pyranose type) X: 2 or 6 or 2.3
or 3.6 (2+3:2:5) (molar ratio). Note that when X has two sites, R has two molecules.

nl: positive integer (iii) P J 2-2 Molecular weight: 170.000, [α), +57. C(c-
0.3, water) Elemental analysis: C36,96%, H6,12$N0.06% Protein analysis: Not detected (Lowry method) Total amount: 90.2% (phenol sulfuric acid method) (Standard material:
Ara, Rha, Gal.

Ga1A) (3:1.5:4.5:1) (-j ratio) Uronic acid analysis: 10.0% (Standard material: Ga1A, carbazole sulfuric acid method) Reduction of carboxyl group (Taylor-Conrad reduction) ) PI3-2-PI3-2R (reduced product) Acid hydrolysis: (IN H2SO4, 100°C, 4h) L-Ara D-Xyl L-Rha L-Fue
D-Glc DG; PI3-2 2.5 1.
0 1.4 0.5 1.0 4.・PI3-2
R2,51,01,40,51,05゜(all molar ratio (Iv) P J 2-3 molecules]・40,000, [α't7 +65.0゜(c
-0.3, water) Elemental analysis: C38, 12%, H6, 32% NO, 08% Protein analysis: Not detected (Lowry method) General FFI: 87.0% (phenol sulfuric acid method) (Standard material: Ara , Rha, Ga 1.Ga IA) (2,1
:1.6:2.1:1. O) (molar ratio) uronic acid analysis:
14.7% (Standard substance: Ga1A, carbazole sulfuric acid method) Reduction of carboxyl group (Taylor-Conrad reduction) P J 2-3-P J 2-3R (reduced product) Acid hydrolysis = (1N H2SO4, 100°C, 4h) L-Ara D-Xyl L-Rha L-Fuc D
-Glc D-GalPJ2-3 3.2 1.0
2. Ili O,50,63,7PJ2-31?
3.2 10 2. G O, 50, 65, 7 (all molar ratios) Structure of 3PJ2-3 [■] R-side chain: -Rha, -Fuc, -Xy 1 partial formula [■
], [■], ■old, gate] = 4 combinations are in any order n6,
n7, n8, n9, positive integer 2) Carbon clearance of polysaccharide component
(1) Experimental Animals After breeding ICR male mice for a certain period of time, 5-week-old mice that appeared to be healthy were used in this experiment as a group of 10 mice.

(2) Experimental method Carbon clearance method of Ilalpern et al. (Ilalpern, B, 11., et al.
al,, J, ReLieulo-endotl+e
1. Soc, I, 77 (1964)), a test solution (PI3-2 and PI3-3 were used as polysaccharide components) was administered intraperitoneally to the above mice at the concentrations shown below, and 24 hours later, carbon suspension was applied. A suspension (Pelican Inc. (manufactured by Pelican) diluted 4 times with physiological saline) was administered into the tail vein at a rate of 5 ml/kg. After 2 and 10 minutes, 20 μl of blood was collected from the mouse's orbit using a micropipette.

After adding 2 ml of 0.1% Na2CO3 solution to the obtained blood to cause hemolysis, 6 ml of 0.1% Na2CO3 solution was added using a spectrophotometer (Kinsei Seisakusho).
Absorbance at 75 nm (OD 675 ) of Al11
The rate of disappearance of the carbon suspension in the blood was determined using the following parameters (control: 1).

Similarly, other test solutions (polysaccharide components: PJN-1 and PJN-3) were also tested (control 2).

(Dosage of test solution) (a) Physiological saline (controls 1 and 2) 5 ml/
kg (b) Polysaccharide component 4II1g/kg -=0.
8mg/ml (5ml/kg) The statistical processing of the results was performed by Studepund.
nt's) to test Nite row Natsuta.

Polysaccharides obtained from bamboobrush carrots, PJN-1, PJN-
FIG. 2 shows the results when 3, P J 2-2, and PI3-3 were administered to mice. All of these carrot polysaccharide administration groups showed a faster elimination rate than the non-administration control group, indicating that the ability to dispose of foreign substances was significantly enhanced.

[Brief explanation of the drawing]

FIG. 1 is a diagram showing the process of separating the target polysaccharide from bamboobrush carrots. FIG. 2 is a graph showing the effect of enhancing carbon clearance ability of the polysaccharide derived from bamboo knot carrots.

Claims (1)

[Scope of Claims] 1. A bamboo-bush carrot-derived product having a molecular weight of about 20,000 to 180,000 and having a constituent sugar selected from arabinose, xylose, rhamnose, fucose, glucose, galactose and galacturonic acid. Polysaccharide. 2. The active ingredient is a polysaccharide derived from ginseng, which has a molecular weight of about 20,000 to 180,000 and whose constituent sugars are selected from arabinose, xylose, rhamnose, fucose, glucose, galactose, and galacturonic acid. Biodefense enhancer.