JP2602295B2 - Polysaccharide, method for isolating the same, and pharmaceutical composition containing the polysaccharide - Google Patents

Description

The present invention relates to novel polysaccharides, in particular calsan A, calsan B, calsan C, calsan D and calsan E, which can be isolated from medicinal carrots or their tissue cultures. Polysaccharides,
The present invention relates to a method for isolating the same and a pharmaceutical composition containing the polysaccharide and having a hypoglycemic effect.

( Prior art) Medicinal carrots, especially Panax ginseng
CAMeyer) has been used as a crude drug for a long time and is known to have anti-fatigue, anti-stress, intestinal, diuretic, and metabolic hyperactivity. It is also used as a treatment for diabetes. Ginsenoside is known as an active ingredient of medicinal carrots. However, for example, there is no known ingredient that exhibits a hypoglycemic effect which is considered to be effective for the above-mentioned diabetes.

(Object of the Invention) An object of the present invention is to find a component having a hypoglycemic action contained in a medicinal carrot. It is another object of the present invention to provide a method for isolating said components. Still another object of the present invention is to provide a pharmaceutical composition containing the component as an active ingredient.

(Constitution of the Invention) The present inventors have repeated studies based on the finding that a water-soluble fraction of a medicinal carrot contains a medicinal component, and as a result, have found a group of novel polysaccharides.

The polysaccharide of the present invention has a hypoglycemic effect and is at least one selected from the group consisting of calsan A, calsan B, calsan C, calsan D and calsan E having the following physicochemical properties, whereby: The above object is achieved: Calsan A molecular weight (gel filtration method): 9.1 × 10 ³ Specific rotation: [α] _D −22.9 ゜ (c 0.14, water) Infrared absorption spectrum (KBr) ν _max (cm ⁻¹ ) : 3440,1070 ¹ H nuclear magnetic resonance (D ₂ O) δ: 4.91 (wide) 5.16 (wide) 5.34 (wide) Elemental analysis (%): C, 37.34; H, 6.41; N, 0.47 Glass Fiber filter paper electrophoresis: 13.0 cm (glucose 11.0 cm) Polyacrylamide gel (30%) electrophoresis: 1.3 cm (bromophenol blue 3.3 cm) Calsan B molecular weight (gel filtration method): 1.4 × 10 ⁴ Specific rotation: [Α] _D -17.6 ゜ (C 0.14, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3450,1060 ¹ H nucleus Magnetic resonance δ: 4.96 (wide) 5.21 (wide) 5.40 (wide) Elemental analysis (%): C, 33.84; H, 4.83; N, 1.34 Glass fiber filter electrophoresis: 16.3 cm (glucose 14.5 cm) ) Polyacrylamide gel (10%) electrophoretic mobility: 1.5 cm (bromophenol blue 3.9 cm) Calsan C molecular weight (gel filtration method): 5.6 × 10 ³ Specific rotation: [α] _D +30.4 ゜ (C 0.12, Water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3400,1050 ¹ H nuclear magnetic resonance δ: 4.94 (wide) 5.19 (wide) 5.36 (wide) Elemental analysis (%): C, 36.75; H, 5.90; N, 2.09 Glass fiber filter electrophoresis: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (30%) electrophoresis: 1.0 cm (bromophenol blue 4.7 cm) Calsan D molecular weight (gel filtration method) ): 1.9 × 10 ³ Specific rotation: [α] _D +21.6 ゜ (C 0.11, water) Infrared absorption spectrum (KBr) ν _max (cm ⁻¹ ) ): 3400,1041 ¹ H nuclear magnetic resonance δ: 5.12 (wide) 5.26 (wide) Elemental analysis (%): C, 35.22; H, 6.14; N, 3.19 Glass fiber filter electrophoresis: 16.3 cm ( Glucose 14.5 cm) Polyacrylamide gel (30%) Electrophoretic mobility: 1.0 cm (bromophenol blue 4.7 cm) Calsan E Molecular weight (gel filtration method): 5.8 × 10 ⁵ Specific rotation: [α] _D -28.8 ゜ (C 0.16, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3450,106 ¹ H Nuclear magnetic resonance δ: 5.15 (wide) 5.35 (wide) Elemental analysis (%): C, 36.59; H , 6.31; N, 0.89 Glass fiber filter paper electrophoresis: 14.5 cm (glucose 14.5 cm) Polyacrylamide gel (30%) electrophoresis: 0.5 cm (bromophenol blue 3.4 cm).

The method for isolating the polysaccharide of the present invention comprises a step of extracting a root of medicinal carrot belonging to the family Ascodae or a tissue culture of medicinal carrot, or a dried product thereof with water and / or an aqueous organic solvent, and an obtained extract. Is subjected to dialysis or ultrafiltration to obtain a calsan mixture containing calsan A, calsan B, calsan C, calsan D and calsan E, thereby achieving the above object.

The pharmaceutical composition of the present invention contains, as an active ingredient, at least one polysaccharide selected from the group consisting of calsan A, calsan B, calsan C, calsan D and calsan E, and has a hypoglycemic effect, Thereby, the above object is achieved.

The polysaccharide of the present invention is a novel compound having the above physicochemical properties and not described in the literature. These compounds are contained in medicinal carrots belonging to the genus Orbaceae or in tissue cultures thereof. The medicinal carrots include Panax ginseng CAMeyer, Panax japonicus CAMeyer, American ginseng ( Panax quinquefolium L.), and Panax ginseng ( Pan ginseng).
ax notoginseng (Burk) FHChen) and Himalayan carrot ( Panax pseudoginseng subsp.himalaicus)
Hara). Of these medicinal carrots, Panax ginseng is particularly preferred. In order to obtain calsan, the root of the medicinal carrot or a dried product thereof (for example, provided as a Chinese herb such as white ginseng or red ginseng) is used. When calsan is obtained from a medicinal carrot tissue culture, the tissue culture is prepared by conventional methods. For example, it is prepared by aseptically culturing a part of the medicinal carrot tissue on a solid medium containing plant hormones to generate calli, and culturing the callus on a liquid or solid medium.

The calsans of the present invention are isolated from the above-mentioned medicinal carrot or its tissue culture, for example, by the following method.
First, the medicinal carrot or its tissue culture is dried if necessary. If necessary, this is degreased using a normal fat-soluble organic solvent, and then extracted with water and / or an aqueous organic solvent. Although the extraction can be carried out sufficiently with water, an aqueous organic solvent may be used to prevent spoilage of the extract and promote the extraction. Also, both may be extracted. As the aqueous organic solvent, a mixed solution of a lower alcohol such as methanol and ethanol, a water-soluble organic solvent such as acetone, acetonitrile, dimethylformamide, and dimethylacetamide and water is used. The concentration varies depending on the type of raw material and the like, but one having a low concentration of about 1% to a temperature of about 60% is used. It is preferable to heat at the time of extraction or to pulverize the medicinal carrot or its tissue culture to be extracted, since the extraction is promoted.

The above extract is subjected to dialysis or ultrafiltration to obtain a calsan mixture. When subjecting to dialysis or ultrafiltration, the above extract may be used as it is or may be used after being concentrated. In addition, after adding a water-soluble organic solvent such as methanol or ethanol to the above-mentioned extract to precipitate a precipitate containing the desired calsan, an appropriate amount of water or the above-mentioned aqueous organic solvent is added to the obtained precipitate. In addition, a dilution may be used.

The calsan mixture thus obtained usually contains at least 60% of the total calsan A, B, C, D and E.
It contains above. It is desirable that the calsan content be 80% or more. Such a calsan mixture may itself be used as a hypoglycemic agent. However, the mixture may be further isolated, for example, by the following fractionation treatment into each calsan component and used as a hypoglycemic agent.

First, the above-mentioned calsan mixture is treated with an anion exchange resin, for example, DEAE-Sephadex (trade name), and fractionated into a fraction containing calsan A, B, C and D and a fraction containing calsan E. . The fractions containing calsan A, B, C and D thus obtained are then subjected to molecular weight fractionation. Examples of the molecular weight fractionation method include a gel filtration method using Sepharose, Sephacryl, Sephadex, and agarose beads (all of which are trade names); a method using an ultrafiltration membrane having a molecular weight cut-off of 1,000 to 100,000. Thus, by the molecular weight fractionation method, a fraction containing calsan A; and calsan B, C
And a fraction containing D. The fraction containing calsan B, C and D was treated with an anion exchange resin (eg, chromatography using DEAE-Sephadex, DEAE-Toyopearl), and further purified using Sephadex-G50 (trade name). Carusan B,
Calsan C and Calsan D are obtained respectively. Carusan A
And the fraction containing Calsan E are further purified by chromatography on an anion exchange resin (eg DEAE-Sephadex, DEAE-Toyopearl).

Calsan A, B, which is a polysaccharide obtained as described above,
C, D, and E were found to have excellent hypoglycemic action as shown in the experimental examples described below, and almost no side effects were observed. Therefore, according to the present invention, calsan A, B, C, D
A polysaccharide which is at least one of E and E as an active ingredient, and has a hypoglycemic effect. The dosage of this pharmaceutical composition varies depending on the disease state, but in the case of oral administration to an adult, 50 to 500 mg per day as calsans (at least one of calsan A, B, C, D and E), Preferably it is 100 to 250 mg. By administering this amount of calsans in two or three divided doses, the desired efficacy can be achieved. Such a pharmaceutical composition can be administered both internally and as an injection. Examples of the dosage form of the internal medicine include powders, tablets including sublingual tablets, emulsions, capsules, teas, granules, and liquids (including fluid extracts, syrups and the like). Pharmaceutical compositions usually contain solid or liquid excipients in addition to the calsans. As the excipient, those known in the art are used, and it is desirable that the excipient is formulated so as to contain calsans necessary for a single dose. Excipients in powders and other powders for internal use include lactose, starch, dextrin, calcium phosphate, calcium carbonate, synthetic and natural aluminum silicate, magnesium oxide, dried aluminum hydroxide, magnesium stearate, sodium bicarbonate, dried Yeast and the like. In addition, transdermal absorbents prepared by adding ordinary excipients are also included in the pharmaceutical composition of the present invention.

(Example) Next, an example of the present invention will be described.

Example (A) Isolation of Callus A part of the tissue obtained from the root of a medicinal carrot was aseptically cultured on an agar medium containing plant hormones to generate calli, and the callus was liquid-cultured. The obtained granular callus (tissue culture) was dried to obtain 20 kg of dry callus. Methanol 40 was added to this dried callus, and extraction was performed three times, once a day. Water 40 was added to the dried callus defatted in this manner, and extracted three times. The extracts were combined and concentrated under reduced pressure to obtain 1.5 kg of extract. Dissolve this extract in water 15 and add ethanol
60 was added to precipitate a precipitate. The precipitate was collected from the precipitate (610 g) and the supernatant (870 g) and dissolved in water. Next, put this in a cellulose tube (Viskin
g, Co; 36/32) for 7 days. 60 g of this inner dialysis solution was applied to a DEAE-Toyopearl 650M column (C1 type; 2.2 cm ID × 40 cm), first eluted with water, and then eluted with a 1 M aqueous NaCl solution. Thus, fraction C-1 containing 800 mg of solids and fraction C-2 containing 2.5 g of solids were obtained.

Fraction C-1 was applied to a Sephacryl S-200 column (4.0 cm inner diameter × 90 cm), and eluted with (0.1 M Tris-HCl buffer (pH 7.0) -0.5 M salt). Thus, fractions C-3 and C-4 were obtained. Fraction C-3 was applied to a DEAE-Toyopearl 650M column (OH type, 2.2 cm
Inner diameter x 40cm; 0.05M Tris-HCl buffer (pH 8.0)-0-1
M salt) to obtain a white powder. This was desalted with Sephadex G-10 (2.2 cm inner diameter x 80 cm), and it was confirmed by gel filtration and electrophoresis that it was a single substance. Examination of the physicochemical properties of this white powder gave the following measurements, which were determined to be calsan A: Molecular weight (gel filtration): 9.1 × 10 ³ Specific rotation: [α _D- 22.9 ゜ (c 0.14, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3440,1070 ¹ H nuclear magnetic resonance (D ₂ O) δ: 4.91 (wide) 5.16 (wide) 5.34 (wide) Elemental analysis (%): C, 37.34; H, 6.41; N, 0.47 Glass fiber filter electrophoresis: 13.0 cm (glucose 11.0 cm) Polyacrylamide gel (30%) Electrophoresis: 1.3 cm (Bromophenol blue 3.3 cm).

Next, the fraction C-4 was applied to a DEAE-Toyopearl 650M column (OH type; 2.2 cm inner diameter x 45 cm; 0.05 M Tris-hydrochloric acid buffer (pH 8.0)-0 to 1 M salt), followed by Sephadex G-50 column. (4.0 cm inner diameter x 60 cm; 0.1 M Tris-HCl buffer (pH 7.0)-0.5 M salt). Three kinds of white powders (I), (II) and (III) were obtained by different elution times. Each of these was separated by Sephadex G-10 (2.2 cm
The solution was desalted through a column (inner diameter x 80 cm) and gel filtration and electrophoresis confirmed that it was a single substance. Examination of each physicochemical property gave the following measured values, (I) for Calsan B, (II) for Calsan C, and (II)
I) was found to be Calsan D: Physicochemical properties of (I) Molecular weight (gel filtration method): 1.4 × 10 ⁴ Specific rotation: [α] _D -17.6 ゜ (C 0.14, water) Infrared absorption Spectrum (KBr) ν _max (cm ^-1 ): 3450, 1060 ¹ H nuclear magnetic resonance δ: 4.96 (wide) 5.21 (wide) 5.40 (wide) Elemental analysis (%): C, 33.84; H, 4.83; N, 1.34 Glass fiber filter paper electrophoresis: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (10%) electrophoresis: 1.5 cm (bromophenol blue 3.9 cm) Physicochemical properties of (II) Molecular weight (gel) Filtration method): 5.6 × 10 ³ Specific rotation: [α] _D +30.4 ゜ (C 0.12, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3400,1050 ¹ H nuclear magnetic resonance δ: 4.94 (wide) 5.19 (wide) 5.36 (wide) Elemental analysis (%): C, 36.75; H, 5.90; N, 2.09 Glass fiber filter electrophoresis: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (30%) electrophoretic mobility: 1.0 cm (bromophenol blue 4.7 cm) physical chemical properties molecular weight of (III) (gel filtration method): 1.9 × 10 ³ Specific rotation: [α] _D +21. 6 ゜ (C 0.11, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3400,1041 ¹ H nuclear magnetic resonance δ: 5.12 (wide) 5.26 (wide) Elemental analysis (%): C H, 6.14; N, 3.19 Glass fiber filter electrophoresis: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (30%) electrophoresis: 1.0 cm (bromophenol blue 4.7 cm).

Next, the fraction C-2 was applied to a Sephacryl S-200 column (4.0 cm ID × 90 cm; 0.1 M Tris-HCl buffer (pH 7.
0) -0.5M salt) and then DEAE-Toyopearl 6
A 50 M column (OH type; 2.2 cm inner diameter × 40 cm; 0.05 M Tris-HCl buffer (pH 8.0) -0 to 1 M salt) was obtained to obtain a white powder. This is Sephadex G-10 (2.2cm inner diameter x 80cm)
And gel filtration and electrophoresis confirmed that it was a single substance. When the physicochemical properties of this white powder were examined, the following measured values were obtained.
Molecular weight (gel filtration method): 5.8 × 10 ⁵ Specific rotation: [α] _D −28.8 ゜ (C 0.16, water) Infrared absorption spectrum (KBr) ν _max (cm ⁻¹ ): 3450 , 1062 ¹ H Nuclear magnetic resonance δ: 5.15 (wide) 5.35 (wide) Elemental analysis (%): C, 36.59; H, 6.31; N, 0.89 Glass fiber filter electrophoresis: 14.5 cm (glucose 14.5 cm) ) Polyacrylamide gel (30%) electrophoretic mobility: 0.5 cm (bromophenol blue 3.4 cm).

Among the above data, measurement of molecular weight, glass fiber filter electrophoresis, and polyacrylamide gel electrophoresis were performed as follows. In addition, C, H,
Components other than N are 0.

1) Molecular Weight Each calsan was subjected to gel filtration using Sephacryl S-200 or 500 to determine the respective retention capacity, and the molecular weight was calculated from a standard curve prepared using Dextran T series.

Sephacryl Calsan A used for molecular weight measurement 9.1 × 10 ³ S-200 Calsan B 1.4 × 10 ⁴ S-200 Calsan C 5.6 × 10 ³ S-200 Calsan D 1.9 × 10 ³ S-200 Calsan E 5.8 × 10 ⁵ S- 500 2) Electrophoresis of glass fiber filter paper Glass fiber filter paper (Whatman GF / C, 15 × 40cm)
The moving distance was measured using.

Conditions: alkaline borate buffer, pH 9.3, 450 V, 2 hours; color development, after spraying p-anisidine sulfate solution, heating at 100 ° C. for 10 minutes.

3) Polyacrylamide gel electrophoresis The moving distance was measured using a column (5 mm inner diameter x 7 cm) packed with 10% or 30% polyacrylamide gel.

Conditions: borate buffer pH 9.3, 2 mA, 2 hours; color development is thymol sulfate method.

(B) Measurement of sugar content and peptide content of calsan The sugar content and peptide content of the above calsans A, B, C, D and E were examined.

Add 2N sulfuric acid to each of Calsan A, B, C, D and E,
The mixture was heated at 100 ° C. for 6 hours to perform hydrolysis. This was reduced with sodium borohydride in water and further acetylated with acetic anhydride in pyridine. 3% ECNSS-M / Gasch
Using a rom Q column (100-200 mesh, inner diameter 3 mm, length 2 m), gas chromatography was performed at 190 ° C with nitrogen (60 ml / min.) to confirm the presence of the following neutral sugars.
The numbers in parentheses indicate the molar ratio present.

Calsan A: rhamnose, arabinose, xylose, galactose (0.2: 0.8: 1.4: 1.0) Calsan B: arabinose, xylose (1.1: 1.0) Calsan C: arabinose, galactose (0.3: 1.0) Calsan D: arabinose, xylose, mannose, Galactose, glucose (1.3: 0.3: 0.4: 1.0: 1.0) Calsan E: rhamnose, arabinose, xylose, galactose (0.1: 0.7: 0.1: 1.0) The following results were obtained when measured by the method.

The following results were obtained when the acidic sugar content was measured by the modified carbazole sulfate method.

Next, when the peptide content was measured by the Lowry method, the following results were obtained.

(C) Blood glucose lowering test using calsan Experimental example 1 (blood glucose lowering test of normal mice) Five normal dd mice having a blood glucose level of 140 to 170 mg / dl and a weight of 25 to 30 g were grouped. The calsans of the present invention were administered intraperitoneally at the doses shown in Table 1 for each of these groups, and the relative blood glucose levels after 7 hours and 24 hours (% relative to the control)
Was measured. Table 1 shows the results. The dialysis solution in Table 1 indicates the dialysis solution of the tissue culture obtained in section (A).

As a result, it is clear that the relative blood sugar was lower in the calsan-administered group than in the control group.

Experimental Example 2 (Hypoglycemic test of alloxan diabetic mouse) Alloxan 35 mg / kg was intravenously injected into the same normal dd mouse as used in Experimental Example 1. After 5 days, blood sugar level is 250 ~
This was 450 mg / dl, which was used as the alloxan diabetic mouse group. In contrast, calsan E, which is the main component of calsans, was intraperitoneally administered at the doses shown in Table 2 and the relative blood glucose level (% relative to control) after a certain period of time was measured. Table 2 shows the results.

As a result, it is clear that calsans also have a hypoglycemic effect in alloxan diabetic mice.

(Effect of the Invention) According to the present invention, the medicinal carrot and / or
Alternatively, calsan A, calsan B, calsan C, calsan D and calsan E, which are novel polysaccharides having hypoglycemic action, can be obtained from the tissue culture. These calsans are easily isolated, have significant pharmacological effects, and have few side effects. Since the pharmaceutical composition containing these calsans can be used as an oral preparation or an injection, it can be widely used, for example, as a therapeutic agent for diabetes.

Claims (8)

(57) [Claims] 1. A polysaccharide selected from the group consisting of calsan A, calsan B, calsan C, calsan D and calsan E, which has a hypoglycemic effect and has the following physicochemical properties: calsan A Molecular weight (gel filtration method): 9.1 × 10 ³ Specific rotation: [α] _D −22.9 ゜ (c 0.14, water) Infrared absorption spectrum (KBr) ν _max (cm ⁻¹ ): 3440,1070 ¹ H nuclear magnetic resonance (D ₂ O) δ: 4.91 (wide) 5.16 (wide) 5.34 (wide) Elemental analysis (%): C, 37.34; H, 6.41; N, 0.47 Glass fiber filter electrophoresis: 13.0 cm ( Glucose 11.0 cm) Polyacrylamide gel (30%) Electrophoretic mobility: 1.3 cm (bromophenol blue 3.3 cm) Calsan B Molecular weight (gel filtration method): 1.4 × 10 ⁴ Specific rotation: [α] _D -17.6 ゜ (C 0.14, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3450, 1060 ¹ H nuclear magnetic resonance δ: 4.96 (wide) 5.21 (wide) 5.40 (wide) Elemental analysis (%): C, 33.84; H, 4.83; N, 1.34 Glass fiber filter electrophoresis: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (10%) Electrophoresis: 1.5 cm (bromophenol blue 3.9 cm) Calsan C molecular weight (gel filtration method): 5.6 × 10 ³ Specific rotation: [α] _D +30.4 ゜ (C 0.12, water) Infrared absorption spectrum (KBr) ν _max (cm ^{− 1} ): 3400,1050 ¹ H Nuclear magnetic resonance δ: 4.94 (wide) 5.19 (wide) 5.36 (wide) Elemental analysis (%): C, 36.75; H, 5.90; N, 2.09 Glass fiber filter paper Electrophoretic mobility: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (30%) Electrophoretic mobility: 1.0 cm (bromophenol blue 4.7 cm) Calsan D Molecular weight (gel filtration method): 1.9 × 10 ³ Specific rotation: [α] _D +21.6 ゜ (C 0.11, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3400,1041 ¹ H nuclear magnetic resonance δ: 5.12 (wide 5.26 (wide) Elemental analysis (%): C, 35.22; H, 6.14; N, 3.19 Glass fiber filter electrophoresis: 16.3 cm (glucose 14.5 cm) Polyacrylamide gel (30%) Electrophoresis: 1.0 cm (bromophenol blue 4.7 cm) Calsan E Molecular weight (gel filtration method): 5.8 × 10 ⁵ Specific rotation: [α] _D -28.8 ゜ (C 0.16, water) Infrared absorption spectrum (KBr) ν _max (cm ^-1 ): 3450,1062 ¹ H Nuclear magnetic resonance δ: 5.15 (wide) 5.35 (wide) Elemental analysis (%): C, 36.59; H, 6.31; N, 0.89 Glass fiber filter electrophoresis: 14.5 cm ( Glucose 14.5 cm) Polyacrylamide gel (30%) Electrophoretic mobility: 0.5 cm (bromophenol blue 3.4 cm). 2. A step of extracting the roots of medicinal carrots belonging to the family Ascidaceae or a tissue culture of medicinal carrots, or a dried product thereof with water and / or an aqueous organic solvent, and dialysis or ultrafiltration of the obtained extract. Carsan A, Carsan B, Carsan C according to claim 1 after filtration.
Obtaining a calsan mixture comprising calsan D and calsan E. 3. The method according to claim 2, further comprising the step of subjecting said calsan mixture to a fractionation treatment to isolate said calsan A, calsan B, calsan C, calsan D and calsan E, respectively. Isolation method. 4. The medicinal carrot is a Panax ginseng ( Pa)
nax ginseng CAMeyer), Panax j
aponicus CAMeyer), American carrot ( Panax qui)
nquefolium L.), Panax notogins
eng (Burk) FHChen) and Himalayan carrot ( Pan
ax pseudoginseng subsp. himalaicus Hara). 5. The extract is concentrated, or a water-soluble organic solvent is added to the extract to obtain a precipitate. The concentrate or the precipitate is dissolved in water or an aqueous organic solvent and then dialyzed. Alternatively, the isolation method according to claim 2, comprising a step of subjecting to ultrafiltration. 6. The simple organic solvent according to claim 2, wherein said aqueous organic solvent is an aqueous solution containing at least one selected from the group consisting of lower aliphatic alcohols, acetone, acetonitrile, dimethylformamide and dimethylacetamide. Separation method. 7. The isolation method according to claim 2, wherein said fractionation treatment is performed by a molecular weight fractionation treatment and an ion exchange resin treatment. 8. The carsan A according to claim 1,
A pharmaceutical composition having a hypoglycemic effect, comprising as an active ingredient at least one polysaccharide selected from the group consisting of calsan B, calsan C, calsan D and calsan E.