JPS5841894A - Sangenone - Google Patents

Abstract

NEW MATERIAL:A sangenone having physical and chemical properties as follows (sangenone D): description: yellow powder, molecular weight: 708 (according to the field desorption mass spectrography), melting point: 175-185 deg.C (decomposed), [alpha]D<25>=-145 deg. (c=0.173 in methanol); positive to ferric chloride reaction and to hydrochloric acid-magnesium reaction, hard soluble in water, hexane, chloroform, soluble in ethyl ether or ethyl acetate, easy soluble in methanol, ethanol, DMF, acetone; acidic substance. USE:Hypotensor with low toxicity. PREPARATION:For example, barks of mulberry tree roots are finely cut, defatted with a low-polarity organic solvent, then extracted with a lower alcohol. The extract is filtered, the filtrate is dried under reduced pressure to obtain an extract. The extract is subjected to column chromatography to separate impurities and collect fractions containing sangenone D. Then, sangenone is isolated and purified.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel compound (Sangenone D) that has a blood pressure lowering effect and is useful as a pharmaceutical.

Until now, mulberry root bark has been referred to as mulberry white bark in Chinese medicine, and has been included in Chinese medicine preparations as an anti-inflammatory diuretic, laxative, and antitussive expectorant.

Recently, the blood pressure lowering effect of methanol extract (
99th Annual Meeting of the Pharmaceutical Society of Japan, Abstracts, 162 pages, 29A
2-3), Hyperglycemic lowering effect of extract (Tokuko Sho 5)
3-44530), interferon-inducing effect of high molecular weight polysaccharide in extract (Japanese Patent Application Laid-Open No. 1983-1999)
99313) etc. are known.

However, regarding the active substance in the blood pressure lowering effect, two types of 8-substituted cyclohexenylflavone derivatives, quanon G and quanon H (Chemical and Pharmaceutical Pretin (Chem. .

Pharm, Bull, ) Vol. 28, No. 8, p. 2548 (1980), Heterocycles (, HETEROC
YCLES, Vol. 14, No. 12, p. 1943 (1980)] have not yet been elucidated.

As a result of intensive research, the present inventors have discovered that the family Moraceae (Moraceae)
tLs) The present invention was completed by extracting plants with an organic solvent, and discovering that a new compound obtained by isolating from this extract had a blood pressure lowering effect, and named it Sangenone D.

First, the method for producing Sangenone D will be explained.

Mulberry root bark is cut into small pieces, degreased with a relatively non-polar organic solvent, extracted with lower alcohol, low Mk fatty acid ester, ether or acetone, filtered, and the P liquid is dried under reduced pressure to obtain an extracted extract. This extracted extract is subjected to column chromatography according to a conventional method to separate impurities, and a fraction containing Sangenone D is obtained. Next, these two fractions are subjected to preparative thin layer chromatography or high performance liquid chromatography to isolate and purify them to produce Sangenone D.

Here, the raw material, mulberry, is used as a commercially available medicine from China such as white bark and magwa (
Moruv alba L,), Yamaguwa (MorLL)
s bambycisKoidz) or Rosou (Mo
rus LhoLLKoidz or Morwmulti
caulis perr, ), etc., and its root bark is particularly preferred. These should be thoroughly dried into fine powders.

Suitable organic solvents with relatively low polarity for use in the degreasing treatment include n-hexane and benzene.

As the lower alcohol for 711, methanol, ethanol, etc. can be used, as the lower fatty acid ester, methyl acetate, ethyl acetate, butyl acetate, etc. can be used, and as the ether, methyl ether, ethyl ether, etc. can be used.

For column chromatography using a conventional method, for example, using a column packed with silica gel, acetone-benzene mixture, methanol-benzene mixture, ethyl acetate-
A fraction containing Sangenone D can be obtained by elution with a benzene mixture or an acetone-hexane mixture.

For the subsequent purification operation, for example, hexane, benzene, ethyl ether, chloroform, acetone, and ethyl acetate are developed by preparative thin layer chromatography on silica gel using a solvent such as methanol alone or in combination, and then preparatively separated. Sangenone D can be obtained by doing this or by repeating this operation.

Note that it is also possible to omit the degreasing treatment during the above extraction/separation operation.

The physical and chemical properties of Sangenone D obtained by the above treatment are as follows.

(a) Properties: Yellow powder (A) Molecule ffi: 708 (from M+ field desorption mass spectrum) (C) Melting point: 175-185°C (decomposition) (d) Optical rotation: [α seat- 145° (C=0.173, methanol)
(e) Infrared absorption spectrum (nu, )'ol ) (c
Fn″″1) (See Figure 1) Size: 3200.16
65 (sh), 1645 (sh), 1620
, (br,), 1555 (sA) (a) Ultraviolet absorption spectrum (njn) (see Figure 2)'λ, 933H (log
g): 220 (Eh, 4.65), 22
6 (, rA, 4.57), 283 (4,33),
288 (Sh, 4.32), 309 (4,30
) λ0H30H+A'. '3(eOgε):225(4,
62), 292 (JA, 4.28), ax 309 (4,39), 352 (Sk, 3.94)
, 400 (, rA, 3.38) (y) Nuclear magnetic resonance spectrum (d6-DMSO1200
MHz, 120°) (711F) δ: 1.37 (3
H,s ), 1.43(3H,,r), 1.67(3H
, s), 1.90-2.20 (IH, m), 2.30
~2.50 (IH,m), 2.59 (IHSddX
J=7.14Hz), 2.89(10, ddXJ=B
, 14Hz), 3.52(I)-1, ddd, J=
4.11.11JIz), 4.26 (IHXbr, d
, J = 11 Hz), 4.78 (IH,t, J = l
lHz), 5.09 (IHlm), 5.16 (IH,b
r, JP), 5.88 (IHXs), 5.96 (
IH, d, J=2Hz), 5.98(l)(, dd,
J=2.8Hz), 6.05 (IHXdd, J=2
．． 8H, 6.14 (IH, dXJ=2H, 6.31
(IHXd, J=2Hz), 641 (kawa, dd,
J=2.8H, 6.76 (IHXd, J=8Hz)
, 7.20 (IH, d, J=8Hz), 7.60 (I
H, d, J = 8Hz) (Gradient color reaction Ferric chloride reaction Positive (reddish purple) Hydrochloric acid-magnesium reaction Positive (orange) Sodium borohydride reaction Positive (orange) (') Molecular formula:
C40H36o12 0) Solubility in solvents 11 Soluble: Water, hexane, benzene, chloroform, dichloromethane Soluble: Ethyl ether, ethyl acetate Easily soluble: Methanol, ethanol, dimethylformamide, acetone (k) Distinction between acidic, neutral, and basic : The strong pressure-lowering effect of the acidic Sangenone D was confirmed by the following animal experiment.

First, the sample was dissolved in ethanol and propylene glycol, and diluted with physiological saline.
Bendoparbital ao m was prepared under anesthesia.
y/kg (su) and urethane 1.5 g/! ? (J?
C) Wistar rats (male, weight 350-4
00g) and spontaneously hypertensive rats (SHR) (JI, weight 200-230g) were administered into the hind limb vein, and blood pressure was measured using a polygraph from the common carotid artery.

As a result, in Wistar rats, a decrease in blood pressure of 35 vmHy was observed at a dose of 1897/9, and in SHR, a decrease in blood pressure of 8011H,9 was observed at a dose of 1 my/kg.

In addition, the LD50 for acute toxicity was determined in mice (5 weeks old, dd
Y strain, male) M intracavitary administration was 300 IIg/kg or more.

As mentioned above, Sangenone D, which has the above-mentioned physical and chemical properties, can be produced from mulberry root bark through a relatively simple extraction and purification procedure, exhibits an excellent blood pressure lowering effect, and is also toxic. It is suitable as an antihypertensive agent because of its low antihypertensive properties.

The method for producing the Sangenone compound (Sangenone D) of the present invention will be specifically explained below.

Production example: 8 kg of shredded dried commercially available medicinal root bark (produced in Guangdong, China) was
Degreased twice with 40/hexane and then twice with 40/benzene at room temperature for one week, then with 40% methanol.
Extraction was carried out twice at room temperature for 3 days each, the extract was collected, the solvent was distilled off under reduced pressure, and ethyl acetate 11 was added.
The soluble portion was separated.

The solvent was distilled off under reduced pressure from this ethyl acetate soluble portion to obtain an extracted extract of 70.9%.

This extract 36.9 was dissolved in the minimum necessary amount of acetone and adsorbed onto silica gel (Wakogel C-2OO, manufactured by Wako Pure Chemical Industries, Ltd.) 7 (L9). After the solvent was removed by evaporation, silica gel 300.9 was filled. Layer packing on top of the column,
First, 10/15 thiacetone-benzene was used as a developing solvent.
After removing impurities by elution, elution was carried out with 20 thiacetone-benzene 5/5, and this eluate was concentrated under reduced pressure to obtain two fractions containing Sangenone D, 113.0, li'.

This fraction was subjected to preparative thin layer chromatography [Wakogel B
-5F (manufactured by Wako Tsumugi Kogyo Co., Ltd.), developing solvent: methanol: chloroform = 1:5, acetone: hexane = 1:1, ethyl ether only (ri next)], and Sandinone I
)t,oy was obtained. Its physical and chemical properties were as follows.

(α) Properties: Yellow powder (h) Molecular weight: 708 (from M+ field desorption mass spectrum) (C) Melting point: 175-185°C (decomposition) (dl specific optical rotation: [α]”-145 °(C=0.173, methanol) (e) Infrared absorption spectrum (nu)'ot)
(cm-') νm(1,z: 320Q, 1665
(tA), 1645 (SA), 1620 (br,), 1
555 (SA) UV absorption spectrum (1L) λOH3°H (C09'): 220 (yA, 4
．． 65), 226 (Sk, 4.57), ax 283 (, -4,33), 288 (, rA, 4.32
), 309(4,30) λOH3°"""' (log ff): 225(
4,62), 292(sh, 4.28), 309(4,
39), 352 (,tA, 3.94), 400(,r
A, 3.38) (y) Color reaction Ferric chloride reaction Positive (reddish purple) Salt-magnesium reaction Positive (orange) Sodium borohydride reaction Positive (orange) (A) Solubility in solvents all : Water, hexane, benzene, chloroform,
Soluble in dichloromethane: Ethyl ether, ethyl acetate Easily soluble: Methanol, ethanol, dimethylformamide, acetone (L) #Distinction between neutrality and basicity: Acidic 0) Elemental analysis value Experimental value (H); C: 64. 75, H: 5.20 (k
) R7 value [Silica gel thin layer chromatography (Wako Tsumugi Kogyo Co., Ltd.: Wakogel B-5F)] 0.31 (Methanomyl: Nyoroform-1:5) 0.61
(acetone:benzene-1:1)

[Brief explanation of drawings]

FIG. 1 is a diagram showing the infrared absorption spectrum (rLtL)ol) of the sungenone compound of the present invention, and FIG. 2 is a diagram showing the ultraviolet absorption spectrum (in methanol) of the sungenone compound of the present invention. Patent applicant Zenyaku Kogyo Co., Ltd. Patent applicant agent Patent applicant agent

Claims (1)

[Claims] 1) Sangenone compound (Sangenone D) 0 having the following physical and chemical properties (a) Properties: Yellow reaction (A1 molecule 1: 708 (from M+ in field desorption mass spectrum) (C) Melting point: 175-185°C (decomposition) (d) Specific optical rotation: Ca] v-145° (C = 0.173, )
(α-1) Shir, Layo: 3200.1665 (zh
), 1645 (sh), 1620 (br, )
, 1555 (sh) (power ultraviolet absorption spectrum (n
) λ0H3°H (log e): 220 (sh, 4
．． 65), 226 (, t=A, 4.57), mayo283 (4,33), 288 (lh, 4.32), 3
09(4,30)' λ2νH+klC1s (e, gε): 225 (
4,62), 292 (, rA, 4.28) 309 (
4,39), 352 (SA, 3.94) 400 (J?
h, 3.38) (y) Color reaction Ferric chloride reaction Positive Salt-magnesium reaction Positive Sodium borohydride reaction Positive (Solubility in gradient solvent SS: water, hexane, benzene, chloroform, dichloromethane ° Soluble: Ethyl ether, ethyl acetate Easily soluble: Methanol, ethanol, dimethylformamide, acetone (t) Distinction between acidic, neutral, and basic: #2) Extract Morus plants with a polar solvent; The sungenone compound according to claim 1, which is obtained by subjecting the compound to silica gel column chromatography and then preparative thin layer chromatography.