JPS5955895A - Soyasaponin - Google Patents

Abstract

NEW MATERIAL:The compound of formula I [R<1> is H or hydroxymethyl; R<2> is H, alpha-L-rhamnopyranosyl or beta-D-glucopyranosyl; R<3> is H or OH; R<4> is H or beta-D-glucopyranosyl (1 3)-alpha-L-arabinopyranosyl]. EXAMPLE:3-O-[ alpha-L-Rhamnopyranosyl( 1 2 )-beta-D-galactopyranosyl( 1 2 )-beta-D- glucuronopyranosyl]soyasapogenol B of formula II. USE:A soybean component effective to prevent the formation of lipid peroxide. PROCESS:Soybean flour is defatted and extracted e.g. with methanol. The extracted active component is concentrated by evaporation to obtained the extract transferred to water-saturated n-butanol. The component is dissolved, e.g. in methanol, precipitated with ether, and separated by column chromatography and centrifugal liquid chromatography.

Description

[Detailed description of the invention] Glycine max Merri
The present invention relates to novel Nyazabonins isolated from ll).

Soybeans are a major source of vegetable protein and vegetable oil, and are commercially processed into soybean flour, soybean oil, soybean protein, and the like. This is due to the fact that soybeans contain a large amount of protein and oil, and other components have also been discovered, such as lecithin and flavones.

The inventors of this discovery further researched the components of soybeans, and as a result discovered new saponins and came up with this invention.Formula (I): [In the formula, 1 is hydrogen LE? - or hydroxymethyl group;
2 is water [atom, α-L-rhamnopyranosyl or β-
D-Guyacobyranosyl group; V is a hydrogen atom or a hydroxy group; R4 is a hydrogen atom or a β-D-glucopyranosyl (l→3)-α-L-arabinopyranosyl group; however, when R1 is a hydrogen atom, R2 is When R2 is a β-D-glucopyranosyl group, 3 is a hydrogen atom. When R2 is a β-D-glucopyranosyl group, the second is a hydroxymethyl group and 3 is a hydroxy group. When V is a hydrogen atom, R4 is a hydrogen atom. When R3 is a hydroxy group. 4 is β-D-glucopyranosyl-(l→
3) -α-L-Arabinobyranosyl group] The compounds and chemical names of the present invention are as follows.

(+) 8-0-(α-L-rhamnopyranosyl(1-
'2)-β-D-galactopyranosyl (l→2)-β-
D-glucuronopyranosyl] soyasapogenol B (abbreviated as soyasaponin I) (tl) 8-0- [a -L-rhamnopyranosyl (l→2) -a-L-arabinopyranosyl (l →2)-
β-D-glucuronopyranosyl] soyasapogenol B
(abbreviated as soya saponin) (Ill) B -0-(β-D-galactopyranosyl (l→2)-β-D-glucuronopyranosyl) Soya zabogenol B (abbreviated as soya saponin) )0 (IV) 8-0-(β-D-glucopyranosyl(1
→2)-β-D-galactopyranosyl(1→2)-β-
D-glucuronopyranosyl)-22-0-[β-D-glucopyranosyl (l→3)-α-L-arabinopyranosyl] Soyasapogenol A (abbreviated as Soyasaponin A1) (v) a-0-(β-D-galactopyranosyl(l→
2)-β-D-glucuronopyranosyl)-22-〇-[
β-1)-glucopyranosyl (1+8)-α-L-arabinopyranosyl] Soyasapogenol A (abbreviated as Soyasaponin A2) The soyasaponins of the present invention can be obtained from soybeans by, for example, the following method. There is a method.

That is, the soybean powder used as the raw material is defatted using a normal fat-soluble organic solvent, the active ingredients are extracted using water, lower aliphatic alcohols, or water-containing lower aliphatic alcohols, and the extract is concentrated by evaporation. Extract. Dissolve this in water-saturated n-butanol, add water to the solution, shake it, let it stand, separate the n-butanol layer, and evaporate to dryness. 1:1) may be dissolved in a mixed suspension and allowed to stand to separate the n-butanol layer, followed by evaporation to dryness. This evaporation residue is dissolved in lower aliphatic alcohol, stirred and poured into ether, and the resulting precipitate is filtered to obtain soybean saponin, which is a mixture of soya saponins.

Another method is to degrease soybean powder, water,
Extract with lower alcohol or hydrous lower alcohol,
The concentrate is dissolved in water or water containing less than about 30% lower alcohol, and this solution is contacted with a macro-network porous cross-linked polystyrene resin adsorbent to adsorb soybean saponin. , lower alcohol or about 80
Soybean saponin is obtained by elution treatment with water containing a lower alcohol of 96 or more.

Furthermore, in order to separate the soya saponin of the present invention from these soybean saponins, the obtained soybean saponin was subjected to silica gel column chromatography [chloroform:methanol:water (f'i 5:85:10, lower layer to 6:4:1).
) mixed solvent] and then centrifugal liquid chromatography [chloroform:methanol:water (10:8:1, lower layer to 7:3:11 lower layer to 14 to 65:85:10, lower layer to 6
:4; ]) mixed solvent] to separate Soya saponin 1.1.1. A and A2 are isolated respectively.

The resulting saponin has been found to be effective in preventing the production of lipid peroxides. This effect is also seen in vivo, and it is involved in lipid metabolism, and can be expected to have various beneficial effects. In addition, the safety factor is that it is a component found in natural soybeans, which are commonly eaten, so it is extremely safe, as evidenced by the results of toxicity tests on mice and rats showing LD, o of 8.2 f/kg or more. .

Next, the present invention will be further explained with reference to Examples.

Example Soya saponin I * J * I m Ale A2
Isolation method: Extract soybean powder (2 to 9) by heating with methanol (101ml) for 5 hours.Filter and extract the methanol extract.
(Methanol-4101 for residual liquid No. 11) was added and extracted by heating for 5 hours. A similar extraction operation is performed three times in total, and the methanol extracts are combined and concentrated under reduced pressure. The concentrated extract was mixed with n-hexane (51
) and extracted twice with defatted methanol extract (16
(lF) is obtained.

Defatted methanol extract (16ON) was distributed into n-butanol-water (]: 1.21), and the water transition part was water-saturated n-butanol-water (1.21).
- Extract with butanol (11).

The n-butanol transition parts were combined and the solvent was distilled off under reduced pressure to give n-butanol.
Obtain a butanol transition extract (50f).

N-butanol transition part extract (50f) was mixed with methanol (
100 fresh) and added dropwise to ethyl acetate (21) to obtain a precipitate (8fi 9).

The precipitate (85F) was subjected to silica gel column chromatography [Merck's Silica Gel G (70-280 mesh) 1
kg; Elution solvent: Chloroform: Methanol-luminium water (65
:85:10. Lower layer~6:4=1) mixed solvent] to obtain total soya saponin ('If) KT-2061 + Elution solvent: Chloroform; Methanol: Water (10:8:1. Lower layer to 7:
3:1. Lower layer -65:85:10. Lower R)~6:4:
1) Mixed solvent], soyasaponin l (3
, 2g).

Soya saponin l (0,5F), soya saponin jaw (0,2F), soya saponin AI (0,8F), and soya saponin A2 (0,7 g) are obtained.

Next, each soya saponin thus obtained has the following properties.

1) mp 288-240°C.

2) [α) D14 B, 0 with an optical rotation of 5° (C = 1.0. methanol) 3) Infrared absorption spectrum (KBr, an) is 8400
(Broad, Strong), 2940, 1785° 1640
．． 4) It exhibits no ultraviolet absorption at wavelengths longer than 210 nm.

5) -fl, G1+ gas resonance spectrum (d5-pyridine.

δ) is 0.70 (8)1), 0.97 (6I[
).

1.29 (80), 1.87 (611),
1.51 (aII) (each singlet), 1.82 (
8H, double line).

6) Soluble in methanol, water, pyridine, dimethyl sulfoxide and ethanol, insoluble in acetone, chloroform, ether, benzene and hexane.

7) It has no odor and its aqueous solution is weakly acidic.

8) Colorless needle-like crystals (crystallized from methanol).

9) Thin layer chromatography using silica gel (pre-coated silica gel 60F254@0.25, manufactured by Merck & Co., developing solvent: chloroform: methanol: water (6
5:85:10. Rf value is 0 if there is an error in the lower layer)
．． 4 is shown.

When a 10% sulfuric acid aqueous solution of 1% cerium sulfate is sprayed on a thin layer chromatograph and heated, it becomes reddish-purple (10) Methane rinse (9)
% hydrochloric acid - heated under reflux in dry methanol) to give 1 mol each of methyl galactoside, methyl glucuronide and methyl rhamnoside.

In addition, when a fully methylated derivative (molecular formula C59Hxoo 01B) obtained by methylation by the Hakomori method (methyl iodide, dimethyl sulfoxide, sodium hydride) is reduced and metathurised, methyl 8,4-diO-methylglucopyranoside, methyl 8 .4. Soak 6-17-0-methylgalactopyranoside, methyl 2,3,4-trilomethylrhamnopyranoside.

Give 1 mole each. In addition, as savogenol, 22
．． 24-di-0-methylsawyasapogenol B (C3
Give 1 mole of 2Hs40B).

Soya saponin 1 1) has an optical rotation of 0 2) [α) D29 + 9.6° (C = 9.5. methanol) with mp 212-215°C.

3) Infrared absorption spectrum (KBr, cm) is 84
00 (broad, strong), 2950.178B.

1 ti 41. , l 050 (Broad, '
-;Qt) has a characteristic absorption maximum at 4) It does not exhibit ultraviolet absorption at wavelengths longer than 210 Bm.

5) 111-4? dda'2LJ #tQx hec) #
(d5-hi+3shin.

δ) is 0.70 (811), 0.95 (611
).

1.19 (911), 1.85 (81-1)
6) Soluble in methanol, water, pyridine, dimethyl sulfoxide, and ethanol, and insoluble in acetone, chloroform, ether, benzene, and hexane.

7) It has no odor and its aqueous solution is weakly acidic.

8) Colorless fine crystals (crystallized from methanol).

9) Thin/li chromatography using silica gel (
Pre-coated silica gel 60 F254.0.25 gate,
Manufactured by Merck & Co., developing solvent: chloroform: methanol: water (65:85:1 (1, lower layer) Rf value 0.
43 is shown.

1% C acid 2 cerium on thin IA chromatograph
When an aqueous solution of lifi acid is sprayed and heated, it becomes reddish-purple.

10) Methyl arabinocytes after metathurisis.

Give 1 mol each of methyl glucuronide and methyl rhamnocyte.In addition, when the fully methylated derivative (Csy H9e 017) obtained by methylation by the Hakomori method is reduced and metathurised, methyl 3,4-di-0-methylarabinopyrano is obtained. One mole each of methyl 3°4-di-0-methylglucopyranoside, methyl 2,8.4-)-o-methylrhamnopyranoside is obtained. Moreover, 22.24-di-0-methylsawyasapogenol B is obtained as sapogenol.

1)'mp 215-216°C.

2) [α]D”, has an l1 but luminosity of +7.9° (C=1.0.methanol).

3) Infrared? ? jJ IJ & yield spectrum (llr, on
) is 8400 (broad, strong), 2955.178
4°164 (1,1080,10150 (broad).

It has a unique absorption maximum.

4) It does not exhibit ultraviolet absorption at wavelengths longer than 210 nm.

5) Soluble in methanol, water, pyridine, dimethyl sulfoxide, and ethanol, and insoluble in acetone, chloroform, ether, benzene, and hydrogen.

6) It has no odor and its aqueous solution is weakly acidic.

7) Colorless crystals (crystallized from methanol).

8) Thin layer chromatography using silica gel (pre-coated silica gel 60 F254.0.25III
*Merck, developing solvent: chloroform: methanol:
Water=65:8fi:10. Rf value 0 when attached to lower layer)
．． 45 is shown.

When sprayed with a 10% biased acid aqueous solution of 1% dicerium sulfate on a thin layer chromatograph and heated, a reddish color appears.

9) Methylgalactoside upon metatolysis.

One mole of methyl glucuronide is obtained in each case.

The fully methylated derivative (C51H86014) obtained by methylation by the Hakomori method is reduced and then subjected to metalysis to produce methyl 2.8.4.6-tetra-
0-methylgalactopyranoside, methyl 3.4-diO
-Methylglucopyranoside is obtained in an amount of 1 mol, and 22.24-di-0-methylsawyasapogenol Bfe is obtained as methylated bogenol in an amount of 1 mol.

1) I[1p is 240-242°C.

2) It has an optical rotation of (ff)D+28.2° (C=0.9.methanol).

8) Infrared absorption spectrum (KBr, Gll'
) is 8B75C broad, strong),, 2918.178
7゜1632, 1885 (broad), 107
4 (broad, strong), 1027 (broad) has a characteristic absorption I box dog.

4) It does not exhibit ultraviolet absorption at wavelengths longer than 210 nm.

5) 13G-nuclear magnetic resonance spectrum (d5-pyridine).

δC) is 172.1 (COOII), 144.0
(C13)e and 108.1.106.5°105.7
．． 104.5.102.9 (Anomeric C)
shows.

6) Soluble in methanol, water, pyridine, dimethyl sulfoxide, ethanol, insoluble in acetone, chloroporum, ether, benzene, hexano.

7) There is no odor, and the aqueous solution is weakly acidic (a solution prepared by dissolving soyazabonin All0''F in 1 liter of water shows pl (5)).

8)%5 colored fine powder crystals (crystallized from aqueous methanol).

9) Thin layer chromatography using silica gel (pre-coated silica gel 60 F254.0.253 & J Merck, developing solvent: chloroform: methanol: water (
6:4:1), the Rf value is 0.25.

1% ceric sulfate and 10% on thin layer chromatography
When the mixture with sulfuric acid aqueous solution is sprayed and heated, WJ#:
exhibit color.

10) Metasurysis (heating under reflux in 9% hydrochloric acid-dry methanol) yields 1 mol of methyl arabinosite, methyl galactoside, and methyl glucuronide and 2 liters of methyl glucoside.

In addition, when the fully methylated product (molecular formula: C7711ia2029) obtained by methylation by the Hakomori method (methyl iodide, dimethyl sulfoxide, sodium hydride) is reduced and metathurised, methyl 3,4-di-0-methylglucopyranoside, methyl B , 4.6-) di-O-methylgalactopyranoside, methyl 2,4-di-0-methylarabinopyranoside and methyl 2,8.
4. Give 2 moles of 6-Citler O-methylglucopyranoside ◎Also, Zavogenol and [,Te2], 24-Di-O-methylsapogenol A (32FN54Q)
Give 1 mole of I) 1) mp is 231-233°C.

2) (a) ”+28.9° (C=0.9.methanol) 1) Has an optical rotation of 3) Infrared absorption spectrum (KBr, cm ) is 83
50 (broad, strong), 2925.1740°16
4-0-1885 (Broad), 1027 (Broad,
has a unique absorption maximum.

4) It shows no ultraviolet absorption at wavelengths longer than 210 nm.

5) 13G-nuclear magnetic resonance spectrum (ds-pyridine) is 172.2 (COOH), 144.0 (C1
3) and 10B, 3.106.0.105.8 and 10
4.7 (hereinafter referred to as Anomeric C).

6) Soluble in methanol, water, pyridine, dimethyl sulfoxide and ethanol, insoluble in acetone, chloroform, ether, benzene and hexane.

7) There is no odor, and the aqueous solution does not show weak acidity (a solution of Soya saponin A25πf dissolved in 1wi of water shows p)16)0 8) Colorless fine crystals (crystallized from aqueous methanol).

9) Thin layer chromatography using silica gel (pre-coated silica gel 60 F254.0.25M shoulder, manufactured by Merck & Co., developing solvent: chloroform; methanol: water =
When applied to 6:4:1), the Rf value did not show 0.3, and the chromatograph showed 1% ceric sulfate and 10%
When a mixture of sulfuric acid is poured and heated, a liquified color appears. 10) Metatrilysis gives 1 mole each of methyl glucoside, methyl galactoside, methyl arabinosite, and methyl glucuronide.

Completely methylated product (obtained by methylation using the Kumo method)
When Ce5lh1oU24) is reduced and metathulysed,
Methylated λ1, - as methyl 3゜4-di-0-methylglucopyranoside, methyl 2,4-diO-methylarabinopyranoside, methyl 2,3.4.6-tetra-0-
Methyl galactopyranoside, methyl 2,8.4゜6-tetra-0-methylglucopyranoside, I7zoi'11
give mol and methylated bogenol, 11.24-
Give 1 mole of di-0-methylsawyasapogenol A.

Claims (1)

[Claims] [In the formula, 1 is a hydrogen atom or a hydroxymethyl group; R2
is a hydrogen atom, a-L-rhamnopyranosyl or β-D-
Glucopyranosyl group, R3 is a hydrogen atom or hydroxy group, R4 is a hydrogen atom or β-D-glucopyranosyl (1-8)-α-L-arabinopyranosyl group; however, 1 (when one is a hydrogen atom, R2 is an α-L-rhamnopyranosyl group, and 3 is a hydrogen atom; when R2 or β-1]-glucopyranosyl group, 1 is a hydroxymethyl group and R3 is a hydroxy group; when R3 is a hydrogen atom, 1 is a hydrogen atom; R3
is a hydroxy group, and 4 is a β-D-glucopyranosyl-(l→3)-a-L-arabinopyranosyl group]. 2. The compound of formula → is B-o-(α-L-rhamnopyranosyl(l→2)-β-
D-galactopyranosyl (l→2)-β-D-glucuronopyranosyl] soyasapogenol B (abbreviated as soyasaponin ■), B-o-(α-L-rhamnopyranosyl (1
→2)-α-L-arabinopyranosyl (l→2)-β-
D-glucuronopyranosyl] soyasapogenol B (abbreviated as soyasaponin l), B-o- (β-D-galactopyranosyl (l→2)-β-D-glucuronopyranosyl) Soya savogenol B (abbreviated as soya saponin ■). 3-o-(β-D-glucopyranosyl (l→2)-β-
D-galactopyranosyl (l→2)-β-Df-lucuronohyranosyl)-22-0-[β-D-glucopyranosyl(,1→3)-α-L-arabinopyranosyl] Soyasapogenol A (abbreviated to Soyasaponin), or 3
-0- (β-D-galactopyranosyl(l→2)-β
-D-glucuronopyranosyl)-22-〇-[β-D-
The compound according to claim 1, which is glucopyranosyl (l→3)-a -L-arabinopyranosyl] soyasapogenol A (abbreviated as soyasaponin).