JPS61197590A - Quassinoid compound and production thereof - Google Patents

Abstract

NEW MATERIAL:The compound of formula I (Glc is glucose residue; R is acetic acid residue; 3,4-dimethyl-4-hydroxy-2-pentenoic acid residue or 3,4-dimethyl-4- acyloxy-2-pentenoic acid residue). EXAMPLE:Bruceantinol 3beta-D-glucoside of formula II. USE:Antitumor agent and antiviral agent. PREPARATION:ATANSHI [seed of Brucea javanica (L.) Merr] is extracted with an alcohol, and concentrated. The obtained extract is partitioned with an organic solvent (preferably chloroform or methylene chloride) and water, and the organic solvent fraction is collected.

Description

Detailed Description of the Invention [Industrial Application Field] The present invention relates to a compound of the general formula (1) (wherein Glc represents a glucose residue, R is an acetic acid residue, and 3,4-dimethyl-4-hydroxy-2 The present invention relates to a quassinoid compound having -A! (representing a ntthenic acid residue or a 3,4-dimethyl-4-acyloxy-2-entthenic acid residue) and a method for producing the same.

Since the compound of the present invention has antitumor activity, it can be used as an antitumor agent.
Future development as an antiviral agent is expected.

[Prior art]

BACKGROUND ART Conventionally, various natural products and compounds derived from natural products that have anti-hazardous effects have attracted attention, and a small number of these have already been put into practical use.

Among these natural products with antitumor effects, Brucea antidysenterica M, which has been used for cancer treatment in Ethiopia since ancient times,
Bruceantin (Bruceantin) is an active ingredient of
Antin) was confirmed to be a compound of the following formula (II).

H (: Kupchan et al. J, Org, Chem,
Volume 38, page 178.

1973; U.S. Patent No. 3,969,369; British Patent No. 1,440,094], and another Bitteraceae plant (Bruce
ajavanica (L,) Merr).

There have already been several reports regarding the components of Ya-dan-Zl [K, H, Lee et al. J, Org.
, Chem, vol. 44, p. 2180.

1979: J, Po1onsky et al. C, R, Aca
d, Set,, Set.

0.268 volume, 1392 pages, 1969; same magazine volume 267.

1346, 1968], and the highly polar fraction and water-soluble fraction have only been reported by the present inventors ("
28th Symposium on Flavor, Terpene, and Seed Oil Chemistry” Abstracts, pages 301 and 304).

[Problems to be solved by the present invention]

Brucea javanica (L,)Me
rr seeds) are known to have antibacterial and antitumor effects. This kumiko is made with pulsator (Bru).
satol (III) (J, Org, Chem, vol. 33, p. 429, 19
In addition to Bruceine (1963)
A, B, C, D.

It is well known that it contains quassinoids such as E, F, G, and H.

The present inventor has discovered a plant of the family Bitteraceae (Simaroubaceae).
) As part of our research, we focused on the highly polar and water-soluble fractions of this mullet. Considering the current situation in which Kumiko is used as a Chinese herbal medicine in China, it is considered that there is an urgent need to study the components of these fractions, which are thought to make a significant contribution to the activity. General formula (1) (wherein Glc represents a glucose residue, R is an acetic acid residue, a 3,4-dimethyl-4-hydroxy-2-pentenoic acid residue, or a 3,4-dimethyl-4-acyloxy- A quassinoid compound having a 2-pentenoic acid residue (representing a 2-pentenoic acid residue) can be produced from Kumiko in the following manner.

That is, after pulverizing the S muntin, it is degreased using a hydrocarbon 1, for example, pentane, hexane, petroleum ether, ligroline, or hebutane, in an amount about three times the amount of the S muntin powder. There are 2 more residues
Extraction is performed using 5 times the amount of alcohol, preferably methanol, at room temperature or under heated conditions. This alcohol extraction is 2
The procedure is repeated three times and the extract is prepared by concentrating the extract.

This extracted extract is mixed with an organic solvent such as ethyl acetate,
By partitioning using an ether-based or chlorine-based organic solvent and water, it is divided into a water-soluble part and an organic layer part. Chloroform or methylene chloride is suitable as the organic solvent used here.

After concentrating the organic layer, add methanol while heating (below 60°C) to make a homogeneous solution. When cooled, the brusatol that separates from door to door, and the p liquid is concentrated again to obtain a dark brown oil. It will be done.

This oily substance can be processed by various chromatography techniques such as adsorption chromatography, partition chromatography, r.p.
It can be purified by a combination of perchromatography or ion exchange chromatography. Purification can also be achieved by combining chromatography and recrystallization.

The pure crystals obtained here were named Yadanzioside I, L and K by the present inventor.

These new quassinoid compounds, yadanzooside I, L, and K, have antitumor effects;
It is expected to be an antiviral drug.

Next, the present invention will be described in more detail with reference to Examples, but it goes without saying that the scope of the present invention is not limited to these Examples.

Example 1 Separation of the organic layer part of the mulberry porridge After pulverizing the mulberry porcelain (60 kJ1), it was immersed in about 1,001 liters of hexane. After performing this operation twice to remove most of the fat and oil, the intestinal mound powder was immersed in 1001 methanol. This methanol extraction was carried out twice, and the methanol was concentrated under reduced pressure. To the resulting syrupy solution, an equal volume of water was added, and further 7 parts of hexane were added. After performing hexane extraction four times to completely defatte the mixture, the aqueous layer was extracted five times with methylene chloride in an amount approximately three times the amount of the aqueous layer.

The extract obtained by concentrating this methylene chloride layer is heated (
(below 60°C), methanol was added little by little to form a homogeneous solution. By leaving this solution at room temperature, the known compound Brusatol precipitated.
) was separated and concentrated again to obtain 1351, a dark brown oily substance.

Examples 2, 4 and 5 describe the method for separating compounds obtained from the organic layer and water-soluble fraction and the physical properties of each substance.
The chromatography conditions used for the separation prior to this are described below.

1. Column A Chromatography using silica Cl (Wako c-200: 85 (1: Wako Tsumugi Pharmaceutical Co., Ltd.). For fraction stores 1 to 6, elution was performed using chloroform: methanol: water (
25:4:1), fractions &7-10 are chloroform:methanol:water (20:4:1), and fractions 11-24 are chloroform:methanol:water (16:4:1'). Using the lower layer, 400 Mt of each fraction was collected.

2. Column B Silica f /L/ (Mallinkrodt 5tlicic
Ac1d; 400, F, Mallinckrodt) to water 26
Partition chromatography with 0- adsorbed. Elution was performed by adding ethanol to chloroform at increasing concentrations of 0.3, 6.9, 12, 15 and 18.21% (11% each).

A fraction of 1 flax and 250 mm was collected.

3. Column C Rover Column Ricloprep RP-8 (Size B,
Chromatography by Merck & Co.). Elution was performed with methanol:water (1:1).

Each ION was collected using a fraction collector.

46 Column D Gluchromatography using Toyonoyaichiru HW-408 (2,4X 100α, Toyo Soda). Elution was performed with methanol, and one fraction of 10/if was collected.

5. Column E Roper Column Ricropred RP-8 (Size B,
Chromatography by Merck & Co.). Elution was performed using methanol:water (3N), and fractions of 10y were collected using a fraction collector.

6. Column F Chromatography using silica gel (Wako C-200, 1.2 kg: Wako Tsumugi Pharmaceutical Co., Ltd.). Elution is frank.

For samples 1 to 12, chloroform:methanol:water (
91.50:12:3) in the lower layer of each fraction. About fractions 13 to 35, about 91 fractions were collected in the lower layer of chloroform:methanol:water (65:35:10).

Example 2 Yadansoside K (sol antinol 3β-D-glucoside) Example 1
Of the 135 g of dark brown oily substance obtained, 3611 was used for one separation and purification.

This dark brown oily substance 36.9% was mixed with silica dal (Wak.

C-200) 75 # and silica chromatograph 7E (Wako C-200, 500
9) Separation was performed.

Fractions 1 to 22. After 11 elutions of each, eluted with ethyl acetate/methanol and fractions 23.
24 (17.6 g) was obtained.

Fractions 23, 24 (17, 6
The following experiment was conducted using 10.7 and F of 1).

10.7y of fraction 23.24 containing glycosides was applied to the above-mentioned column A, and fraction shop 14 (958~)
was further applied to column B. This fraction 416.17 (524 η) of column B was then applied to column C, and the obtained fractions 17 to 24 (291 η) were separated using column D, and the 7 fractions A41 and 4
The title compound t-24119 was obtained from 2.

Physical properties IR spec) #(KBr, 3-'): 3450,17
40,1645°W spectrum ψ (ethano/'):
221.6 nm (g 16500) 252.4n
m(t 8630) [α), , +15 (C=1.0, -1-ta/-
#) Melting point: 214.6-216.5°C (recrystallization solvent: methanol) High resolution mass spectrum (C28H3401
) Calculated value: 546.2100 Actual value: 546.2100 KIffs (m/z): 546 (M"-162-60
), 109.60 (B, P) NMR spectrum (C3D
5N, 90MHz): δ1.42 (3H, s)
1.47 (3H, s), 1.68 (3H, a), 1.9
8 (3H, s).

2.04 (3H, brg), 2.26 (3H, s), 3
．． 89 (3H, s).

6.06 (IH, a), 6.85 (LH, d, J=1
3Hz) Example 3 After pulverizing mulberry kumiko (60kf), it was immersed in about 1004 hexane. After performing this operation twice to remove most of the fats and oils, the Kumiko powder was immersed in 1002 methanol. This methanol extraction was carried out twice, and the methanol was concentrated under reduced pressure.An equal volume of water was added to the resulting syrupy solution, and further 7-e of hexane was added. After performing hexane extraction four times to completely defatte the mixture, the aqueous layer was extracted five times with methylene chloride in an amount approximately three times the amount of the aqueous layer. After separating the methylene chloride layer, the aqueous layer was concentrated under reduced pressure to approximately 1.0
The water-soluble fraction of kII was obtained as a dark brown oil.

50 g of this aqueous layer (1.0 kg) was attached to the above-mentioned column FK.

Example 4 Yadandioside ■ (Prusein B, 3β-D-glucoside) When 2-lactone 16 of column F of Example 3 was concentrated, the known brucein E was reduced to 1.20. @Obtained.

This was separated and the remaining mother liquor (1,159,9) was then separated by column D.

Fractions 44 to 48 in column D were collected and concentrated to give 192Iv of the title compound.

Properties: Colorless needle crystals (recrystallization solvent: ethanol) Melting point:
287-290°C (decomposition) [α), -21° (C = 1.0.
1650°1620.1065 Old spectrum (ethanol): 254nm (g9700
) NMR spectrum (C5D5N, 90 MHz):
δ1.69 (3H, d) 2.04 (3H, a), 2.1
0 (3H, s), 3.81 (3H, a).

6.80 (IH, d, J=13Hz) High resolution mass spectrum” Calculated value as 23H28o11: 48
0.1631 (M-C6H1゜05) Actual value: 480
．． 1646 Example 5 Yadandioside L (2lucein C3β-D-glucoside) Prusein E (450J9) obtained by concentrating Araxin, N17, and N8 in column F of Example 3 was taken from house to house, and the remaining mother liquor (2
700 Da) by column DK, and the fraction 3
5-40 (935 da) was further applied to column E.

From fractions 43 to 56 of column E, 192 pieces of the title compound were obtained.

Properties: Colorless needle crystals (recrystallization medium: methanol/ether) Melting point: 199-204°C [α-knee 0.7° (C=6.2.methanol) IR
Spectrum (KBr, cIn-'): 3420.173
5.1640.

Takumi Suiktor (methanol): 220nm (ε1400
0) 254nm (#9000) NMR spectrum (C3D5N, 90 MHz)
: δ1.43 (6H, s).

1.69 (3H, a), 2.03 (3H, s), 2.3
8 (3H, +).

3.0-3.3 (2H), 3.71 (3H, s), 5.
40 (IH, d.

J=7Hz), 6.70 (IH, s), 6.85 (IH
, d, J=13Hz) Calculated value as 28H36o12: 564.2205 (M-C6H,,0,2'
) Actual value: 564.2182 [Effects of the invention] Yadandioside 1. Ni and L have superior antitumor activity against murine leukemia P388 at doses of pruseoside A, a known compound, and 5 to 101 J9/kIi. From this, these compounds are expected to develop in the future as new antitumor and antiviral drugs.

Claims (3)

[Claims] (1) General formula ▲ Numerical formula, chemical formula, table, etc. ▼ (In the formula, Glc represents a glucose residue, R is an acetic acid residue, a 3,4-dimethyl-4-hydroxy-2-pentenoic acid residue or 3,4-dimethyl-4-acyloxy-2-pentenoic acid residue). (2) Claim 1 in which R is an acetic acid residue, a 3,4-dimethyl-4-hydroxy-2-pentenoic acid residue, or a 3,4-dimethyl-4-atoxy-2-pentenoic acid residue
Compounds described in Section. (3) Brucea javanica (L.
) Merr seeds) are extracted with alcohol, and then the organic solvent fraction is collected and purified. General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, Glc represents a glucose residue. , R represents an acetic acid residue, a 3,4-dimethyl-4-hydroxy-2-pentenoic acid residue, or a 3,4-dimethyl-4-acyloxy-2-pentenoic acid residue) .