JPS62207288A - Rosin glucoside and production thereof - Google Patents

Abstract

NEW MATERIAL:The compound of formula (R1 is H or isobutanoyl; R2 is H, isobutanoyl, 2-methyl-3-hydroxybutanoyl or alpha-methylbutanoyl). USE:An evacuant. PREPARATION:Root of Ipomoea orizabensis Ledanois is crushed, extracted with an organic solvent preferably at normal temperature for 1$10 days and, if necessary, defatted with hexane, etc., and the solvent is distilled off. The extracted fraction is subjected to normal-phase column chromatography and then gelfiltration to remove foreign materials. The product is subjected further to normal-phase chromatography to obtain several fractions having different R1 and R2, which are purified by high-performance liquid chromatography.

Description

DETAILED DESCRIPTION OF THE INVENTION Background of the Invention The present invention relates to novel resin glycosides and methods for their production. This compound is derived from the plant root Ipomoea orllza.
- It can be extracted and isolated from the tuberous root of Benala Ledanoia (hereinafter referred to as Oryzaba).

PRIOR ART Morning glory (Pharbitl@N order (L,)) belonging to the Convolvulaceae (Oonvolvulaaeae) plant ch
olay ), Ipomoaa purga Hayr
r@(Exogonlum purga(Weuder)
,) Beuth,), Ipomoea oriz
abenala (Pe1let, ) Ledanoi
The seeds or tuberous roots of A, etc., have been used as a laxative since ancient times. ) Its cathartic components include pharbltln and pharbltln.
(Jalapin), orizabine (orizabln)
) and other resin glycosides are known.

The above-mentioned resin glycosides are all polymers and are a mixture of several types with similar structures and properties, the constituent organic acids are diverse, and the ester bonds in the molecules are unstable. Isolation and chemical structure studies of many substances have been very difficult.

In 1993, Mannioh et al.
aald Ipurolla acid,
It has been proposed that the aglycone is a compound in which several organic acids are bonded to the sugar hydroxyl group of an oligoglycoside (glycoside acid) (Aroh, Pharm, etc.).
, , 2ヱmutual, J// (/ri3j)).

After that, various reports disappeared [Tetrahedron Letters (Tetra'hem on L@tters)
), Mu 3/23 (/ri 70), f3. European Journal of the Pharmaceutical Society of Japan (
ahmlcalPharmaoutlaal Bu
ll@t1n), 20.114' (lri 7), Nobel, Yuyo, Ko 3! (/ri7J)
, Phytochemistry (Phytoch@m1-at
ry, ヱL, Tata 7 (/ri 7 Jr)), all of which are concerned only with constituent organic acids or glycosides, and the resin glycosides themselves have not yet been elucidated.

[Summary of the invention]

Summary The present invention relates to a resin glycoside having a cyclic ester structure in the molecule.

That is, the resin glycoside according to the present invention has the following formula (11% formula %).
The invention is characterized by obtaining resin glycosides from the tuberous roots of the plant.

H5 Effect The resin glycoside according to the present invention belongs to the Convolvulaceae (Convolvulaa@aa) plant (Ipomoea orzabansla), which is used as a purifying herbal medicine.
) Since it is obtained from the tuberous root of Ledanota (Oryzaba root), it has great potential as a laxative (laxative).

In addition, these resin glycosides are new substances that have been isolated and purified for the first time (F), and are expected to have the following pharmacological effects.

The resin glycoside in the present invention is a compound represented by the above formula (1).

Specific examples of these compounds along with their properties are as follows.

"J1 Arashi Ω" Mouth 1 Compound name Type of substituent R2==hydrogen atom R2 = imbutanoyl group R2=comethyl-3-hydro xybutanoyl group Properties (a) Oc-/ (1) Appearance: Colorless needle-like crystals (2) Specific rotation: [α)Dz, J.

(3) Melting point: //4 cm//℃ (4) 1H-NMR (Pyridine d6,3 tata, 63
M output) See Table-7 below (5) FD--MS 2 m/z //2 (
M+Na)” (6) Negative TAB-MS:
m/, /10! (M-H)-(B) OC-λ (1) Appearance: Colorless acicular crystals (2) Specific rotation: [α]n-jj' (3) Melting point: //J'-/2/"C (4) 1
H-NMR (pyridine d9,3tata, jjMHz) See below (5) FD-MS: m7'z / / 4'j
(M + Na)” (6) Negative FAB-MS
: V2///ri(MH)-←→ 0C-3 (1) Appearance: Colorless acicular crystals (2) Specific rotation: [α]D-2,7° (3) Melting point: /1lI- 772℃ (4)')I-NM)L(pyridine d9,3tatajjM
(5) FD-MS: m/z//2 (M +
Na ) + (6) Negatia”TAB-MS:
mA/10j (MH)-(→ OC-≠ (1) External: Colorless acicular crystals (2) Specific rotation: [α].-13,3° (3) Melting point +
//r-723℃ (4) H-NMR (pyridine 9,3 tata, lt(
5) FD-MS7 m/z //! Y(M+Na
)”(6) Negatia”FAB-MS:mA //
J! The (M-H)-hydrogenated compound is I, ortza, which belongs to the Convolvulaceae family and is used as a folk medicine.
Since it is the main component of the tuberous roots of b@nal Hatake Le-danoia, its toxicity is thought to be extremely low.

Obtaining a Compound The compound of formula <1) can be obtained by any suitable method or means. Because of its complex structure, extraction from Oryzaba roots is preferable.

Oryzapa root Oryzapa root is a plant belonging to the Convolvulaceae family that lives in eastern Mexico.
pomoea orlzabensia (
Pe1let,) The tuberous root of Ledanois, its resin component resin glycosides, is widely used as a laxative together with resin glycosides of Kengo roe, Yarappa root, etc. (
The book ``Herbal Pharmacology'' edited by Michi Fujita, published by Nanzando (l.62), Planta Madlaa (ヱ,, /≠j (/l.6/)).

When obtaining resin glycosides from the above-mentioned Oryzaba root, the material part can be any part of this plant, such as the vine, but preferably the shredded tuberous root, which can be dried or used for extraction as is. be able to.

Although the extraction of the target substance can be carried out in any suitable manner, one preferred embodiment consists of the following steps.

(b) Extraction Crush oryzaba roots into appropriate sizes and extract by immersing them in an organic solvent.The organic solvent to be used as an extractant can be any organic solvent.Preferably, the organic solvent has a carbon number At least one organic solvent selected from lower aliphatic alcohols (preferably -hydric alcohols), ethyl acetate, acetone, ethyl ether, chloroform, and dichloromethane can be used.

Extraction can be performed either under heating or at room temperature, but extraction time is long at room temperature (several days to several weeks), and short at heating (J to 70°C) (several hours to several weeks). Needless to say, the temperature should be kept at room temperature for 7 to 10 days.

Further, if necessary, stir or
10th Edition Japanese Pharmacopoeia Explanation, A, B, A-/ko/ (/
The extraction efficiency can also be increased by PI/ )).

(Cut Purification) After degreasing the above extract with hexane, petroleum ether, etc. as necessary (this degreasing operation may be performed in advance before subjecting the plant to extraction), the extraction solvent is distilled off ( (Usually, this is carried out under reduced pressure at 60° C. or lower).Then, the resin glycoside fraction can be obtained by purifying by normal phase column chromatography and then by gel filtration to remove impurities.

Here, 5-phase column chromatography is performed using silicic acid,
Column chromatography using a highly polar resin solid phase carrier such as silica gel, and the mobile phase is usually
A solvent that is less polar than the same phase carrier can be used, such as a mixture of a nonpolar solvent such as chloroform, dichloromethane, hexane, or ethyl acetate, and a lower alcohol containing water or not. As a specific example of the above method, as shown in Examples below, silica gel chromatography [developing solvent: chloroform-methane (10
: / v/v ) ”].

On the other hand, gel filtration is a method for separating water-soluble polymeric substances with different molecular weights using a polymeric gel filtration agent that has a three-dimensional network structure with a uniform pore size. As a gel filtration agent, dextran gel (e.g. 5ephada:t
■(Pharmacla Fine Che-rolc
ala)), polyacrylamide gels (e.g. old o-Ge1 (Bio-Rad Laboratory
ea)) or agarose gel polystyrene-divinylbenzene copolymer, etc., may be used, but preferably, a gel filtration agent made of a new oil-based polystyrene-divinylbenzene copolymer is used. As a specific example of the above method, polystyrene-divinylbenzene copolymer (Bio-Bea
It can be purified using a dsS-X2 column [solvent: benzene-ethyl acetate (/:/'/v))].

The resin glycoside fraction obtained by the above method was further purified by normal phase column chromatography to a fraction that showed an almost single spot on TLC, and then purified by high performance liquid chromatography (HPLC). Glycoside Oc-/
~Oc・-da can be isolated in pure form.

Note that the normal phase column chromatography referred to here is the same as described above, but as a specific example of the developing solvent, a water-saturated chloroform-methanol mixture (Li: / v/v %) is used as shown in the example below. Purification was performed using

Further, high-performance liquid chromatography may be of any type, but preferred is Bunparatib HPI, C with a high theoretical plate number (/~!0,000).

Experimental Example (1) Isolation of Oa-/~Oc-Hiroshi Dried oryzaba root (roog) was ground into a fine powder, and n-hexane (
1,! liter, 7 days), CHCj5 (j liter,
3 consecutive days), MeOH (/, j liters, 1 day)
(Most of the resin glycosides are transferred to the CHCl5 soluble portion.) C
Concentrate the HCl5 extract under reduced pressure (below 60°C) to obtain CHC.
15 extract (λ) 3 g) was obtained.

This material was subjected to about 9 times the amount of silica gel column chromatography ("Merck Art 77317 Kiear g@l 60
J (manufactured by Merck & Co.).Solvent: CHCl5-MeOH
, 10:/v/v) to obtain a crude resin glycoside lactone (770 g).
After decolorization and purification by passing through a Beada 5X-j column (manufactured by Bio-Rad) (solvent: benzene-EtOAc/s/V/v), a Rover column (Merck Silkagel, solution g + CHCl5-MeOH, reagent: / v/
v) on the TLC.G Pr@packe
d sillaagel (Kusano Science), 2φ×30
(B×2, solvent: CHCl5-MsOH-H2O(
F i /: 0. /'7v%) Flow rate: j,!
The compound Oc- /~Oa-μ tomg, uOmy, respectively
, tsuzo mtp, was isolated with a yield of 3. Yield from oryzaba roots, o, t3 superior, s, yt s, 3) 1%
, 1. It was [O].

Confirmation of structure The resin glycosides of orlzabi roots are converted into orlzabic acid (orlzabi acid) by saponification as shown in Table 7 below.
a acid) and organic acids (Abstracts of the Octopus Annual Meeting of the Pharmaceutical Society of Japan, λ≠! (/ri72)
).

In addition, in the mass spectrometry of Oa-/~Oc-Hiro, as mentioned above, the FD-MS spectrum shows (M+ Na)'',
In the negative FAB-MS spectrum, ions assigned to (NH)- are observed, respectively.
Oc-/~Oc-φ is a combination of organic acids (isobutyric acid, α-methylbutyric acid, nylic acid) to oryzavic acid C (hereinafter referred to as Oa-aatd), and further aglycone yarabinolic acid. It is suggested that there is a cyclic ester bond.

In addition, "HNMR of Oa-/~Oc-φ and Oa-2 peracetate and Oa-aald in deuterated pyridine
By comparing the spectra, the sugar moiety bonding position of the organic acid was determined to be O
6th position of glucose at a-/, 2.3rd position of rhamnose, 3rd position of quinovose, 4th position of glucose at Oc-1 to Oc-1
Ranked 2. Rhamnose. The j-position is estimated to be a kinobose site (see Table 2). The symbols in the table have the following meanings.

☆: Signal overlaps with other signals [unconfirmable☆☆: ddd(/l, 0, ta, 0.!
, 0) Root proton of underlined nearsiloxy group qul: quinovose rba: rhamnose fuc: fucose jla: yarabinolic acid tga: tiglic acid nla: nylic acid lba: isobutyric acid mba iα-methylbutyric acid Ac: acetic acid, then Oc-/ The bonding site of each acyl group in ~0c-44 was determined using an acyl group rearrangement reaction during alkali treatment. That is, 0c-2 to Oa-'A was left in a triethylamine-methanol (/:j) solution at room temperature for 3 minutes, and the resulting product was structurally analyzed using 1H-NMR. As a result, Oc-/~0c-4' has a common substructure (glc-nlaSrha-tga, rba5-
jla,quill-OHa (This partial structure corresponds to the compound of formula (1) above in which R1 and R2 are both H, so this will be referred to as compound A.Glc nigercose, It was revealed that rha: rhamnose, qul: quinovose, nla: nilic acid, tga: tiglic acid, and jla: yarabinolilic acid are bound.

Next, Compound A was investigated using deacylation reaction and acyl group rearrangement reaction in alkali treatment (
Compound A with 7thiammonia (acetone-water (/:/))
The reaction was carried out in a solution at room temperature for 3 hours, and the resulting product was
The structure was analyzed using H-NMR. As a result, compound A
It was suggested that nilic acid is bound to the 6-position of glucose, and tiglic acid and lactone are bound to the 2°3-position of rhamnose.

However, for rhamnose in which both the 2- and 3-positions are acylated, deacylation of one may be accompanied by rearrangement of the other, so the ozonolysis of 0c-2 (-/g
``C, in methanol) to confirm that no rearrangement occurred.

Furthermore, in 1H-NMRVc of compound A, rhamnose is considered to have an unstable conformation close to C1fil K, so it is assumed that rhamnose is in a deacylated form of compound A (niliku acid is removed from the 6-position of glucose). Released compound) Ri! Heating in timethanol for 30 minutes yielded a compound in which the lactone bond was quantitatively rearranged to the 2-position, with no reverse rearrangement occurring. This revealed that tiglic acid was bound to the 2nd position of rhamnose, and lactone was bound to the 3rd position.

From the above results, the structure of Oc-/=Oc-4' was determined.

Claims (1)

[Claims] 1. A resin glycoside represented by the following formula. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R_1 is a hydrogen atom or an isobutanoyl group, and R_2 is a hydrogen atom, an isobutanoyl group, a 2-methyl-3-hydroxybutanoyl group, or an α-methylbutanoyl group. 2. Ipomoea oriza-bens
A method for producing a resin glycoside represented by the following formula, which comprises obtaining the resin glycoside from the roots of Ledanois. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [where R_1 is a hydrogen atom or an isobutanoyl group, and R_2 is a hydrogen atom, an isobutanoyl group,
3-hydroxybutanoyl group or α-methylbutanoyl group] 3. The production method according to claim 2, wherein the method for obtaining resin glycosides from Oryzaba root comprises the following steps. (a) Subjecting Orizaba roots to extraction with an organic solvent. (b) Subjecting the resulting extracted fraction to normal phase column chromatography and then purification by gel filtration. (c) The resin glycoside fraction obtained from the above step is subjected to normal phase column chromatography to be fractionated into different R_1 to R_3. (d) The resulting resin glycoside is further purified by high performance liquid chromatography.