JPS6287591A - Novel quassinoids and their preparation - Google Patents

Abstract

NEW MATERIAL:A compound of formula I [R is residue of 3,4-dimethyl-2- pentenoic acid, residue of 3-ethyl-methl-4-acyloxy-2-pentenoic acid; formula II is formula III-V (Glc is residue of clucose residue; Y is H, glucose residue); where when R is 3,4-dimethyl-2-pentenoic acid residue, formula II is a group other than formula III and formula IV]. USE:Antitumor agent, antiviral agent. PREPARATION:Seeds of Brucea Javanica (L.) Merr are extracted with alcohol to collect organic solvent fractions and purified or the extract is collected, purified and subjected to degluconization reaction.

Description

Detailed Description of the Invention [Industrial Application Field] The present invention relates to a compound of the general formula (I) It represents a 4-acyloxy-2-pentenoic acid residue (where Glc represents a glucose residue and Y represents a hydrogen atom or a glucose residue).

However, when R is a 3,4-dimethyl-2-pentenoic acid residue, the present invention relates to a novel quassinoid compound having a group other than H3 (where Glc is as defined above) and a method for producing the same.

More specifically, since the compound of the present invention has antitumor activity, it is a compound that is expected to develop in the future as an antitumor agent and an antiviral agent.

[Prior art]

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

Among these natural products that have antitumor effects, Brucea antidysenterica M. has been used for cancer treatment in Ethiopia since ancient times.
Pruseanthin (Bruce
ar+tin) was confirmed to be a compound of the following formula (II).

S□ "・ (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
Regarding the ingredients of Yadan-Zi, which is the seed of A. javanica (L.) Merr,
There have already been several reports (K, H, Lee et al. J, O
rg, Chem, vol. 44, p. 2180, 1979
Year; J, Po1onsky et al. C, R, Acad,
Sci, Ser, C+ vol. 268, p. 1392, 1969; same magazine vol. 267, p. 1346, (
1968], and its highly polar fraction and water-soluble fraction Q
The inventors have only reported on
F "28th Symposium on Flavor, Terpene and Essential Oil Chemistry" Abstracts, pages 301 and 304 and [Chemical Society of Japan 50th Spring Annual Meeting J Lecture Proceedings ■ pages 87 and 888]
.

Problems to be solved by the present invention] Brucea javaniea (L,) M
err seeds) are known to have antibacterial and antitumor effects. This gall child has brusatol (Br)
USA to l): ■) υ J, Org, Chem, vol. 33, p. 429,
1963)〉Also Bruceine
A, B, C, D.

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

The present inventor is a plant of the family Bitteraceae (Simaroubaeeae).
) As part of this research, we focused on the highly polar and water-soluble fractions of this gall gall. Considering the current situation in which lily gall is used as a herbal medicine in China, it is considered that there is an urgent need to research the components of these two components, which are thought to make a major contribution to the activity.
We conducted research to elucidate the ingredients.

[Means for solving problems]

The novel quassinoid compound of the present invention having the general formula (I) can be produced from F. oleracea in the following manner.

That is, after pulverizing the Iq gall powder, it is defatted using a hydrocarbon, such as pentane, hexane, petroleum ether, ligroline, or heptane, in an amount approximately three times that of the Iq gall 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 performed two or three times, and the extract is prepared by concentrating the extract.

This extracted extract is mixed with an organic solvent such as an acetate ester,
By partitioning using an ether-based or chlorine-based organic solvent and water, the water-soluble portion and the organic layer portion S are separated. Chloroform or methylene chloride is suitable as the organic solvent used here.

After concentrating the organic layer, add methanol while heating (below 60°C) (7) to make a homogeneous solution, filter out the brusatol that separates by cooling, and concentrate the filtrate again to form a dark brown oil. can get.

This oily substance can be purified by a combination of various chromatographies, for example adsorption chromatography, partition chromatography, gel filtration chromatography or ion exchange chromatography. Purification can also be achieved by a combination of chromatography and recrystallization.

The pure crystals obtained here were named Yadanzioside N and O by the present inventor.

This Yadandioside N is methyl 13.20-epoxy-
2-o-β-D-glucoside 11β, 12α-dihydroxy-15-((3,4-dimethyl-1-oxo-
2E-pentenyl)oxy)-Li2-thioxobiclas-2-ene-21-oate.

On the other hand, Yadandiosito O is methyl 13.20-epoxy-
2-o-β-D-glucoside 11β, 12α-dihydroxy-15-C (4-acetoxy-3-ethyl-4
-methyl-2E-pentenyl)oxy]-3,16-thioxobiclas-1-ene-21-oate.

These novel quassinoid compounds, Yadandioside N and 0, undergo a deglucose reaction by hydrolysis with an acid or an enzyme, and after the reaction, pure crystals can be obtained by using the above-mentioned purification method.

Here, the acid is preferably hydrochloric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, etc., and the enzyme is preferably β-glucosidase.

The present inventor named the crystals obtained here as Yadandioside N-aglycone and Yadandioside O-aglycone.

The novel quassinoid compound of the present invention having general formula (I) has an antitumor effect and is therefore antitumor.

It is expected to be an anti-inflammatory drug.

EXAMPLES 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 of Togo bile (60 kg) After crushing Tochi bile (60 kg), approximately 1,007 g. It was immersed using Nohehonsan. After performing this operation twice to remove most of the fat and oil, the Tomoko powder was immersed in ioog methanol. This methanol extraction was performed twice, and the methanol was concentrated under reduced pressure to obtain a syrupy solution, to which an equal volume of water was added, and further 7β hexane was added. -. After the xane extraction was performed four times to completely defatte the mixture, it 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 filtered off and concentrated again to obtain 135 g of a dark brown oily substance.

In Examples 2 and 3, the separation method and physical properties of Yadandioside N and 0 will be described, but prior to this, the chromatography conditions used for separation will be described below.

Column A Silica gel (Wako C200; 2.5 kg
i Chromatography using Wako Pure Chemical Industries, Ltd. Elution was performed using the lower layer of chloroform:methanol:water (25:4:1) for fractions 1 to 10, and the lower layer of chloroform:methanol:water (25:4:1) for fractions 11 to 30.
Using a lower layer of chloroform:methanol:water (20:4:1), 500n/2 of each fraction was collected.

Column B Roper Column Ricrobrep RP-8 (Size C,
Chromatography by Merck & Co.).

The mixture was extracted with methanol:water (4:6 to 1:1), and 10 g each was collected using a fraction collector.

Column C Toyopearl HW-4OS (2,4X 100an,
Gel chromatography using Toyo Soda). The elution is
This was carried out using methanol, and 10 g of each fraction was collected.

Column D Rover column Ricrobrep RP-8 (size B,
Chromatography by Merck & Co.).

Elution was performed with methanol:water (4:6 to 1:1), and 10 g portions were collected using a fraction collector.

Example 2 Yadandioside N ♀H HOOHCHx 122 g of the extract obtained in Example 1 was used for separation and purification.

122 g of this extract was transferred to silica gel (Wako CC-2
00) 260 and separated by silica gel chromatography (Wako C-200, 2 kg).

Fractions 1 to 30 with Hensen-ethyl acetate, 1.
After eluting 51, 35 to 37.55.3g of fractions containing glycosides were eluted with ethyl acetate/methanol.
I got it.

This was coated with silica gel (Wako C-200) 10
0g and subjected to silica gel chromatography (column A), fractions 26 and 27 (2,
7g) was further separated by column D. From these fractions 70 to 76, 45 parts of the title Yadandioside N were obtained.

Melting point: 175-180℃ [α) ”: +8 @ (CC1,8, ethanol) I
R spectrum (KBr, cm-'): 3420, 1
740.1680.1640.1080.1045.1
020UV spectrum (ethanol, λmax): 2
22 nm (ε=13000), 255 nm (t=
5000) high-resolution mass spectra (CzallziO
) Calculated value: 548.2258 Actual value: 548.2264 NMR spectrum (Pyridine-ds+δ): 0.85 (
6)1. d, J=7.0Hz), 0.87(3fl,
d, J=8.81(z)1.87(3H,s), 2.
18 (3B, s), 3.77 (311, s) 4.90
(LH, brs), 5.12(II(, dd, J=7
．． 7Hz) 5.13 (IH, brs), 5.47 (l
tl, d, J = 7.0IIz) 5.89 (LH, s),
6.10 (IH, d, J = 2.2Hz) 6.20 (I
II, d, J = 4.4 Hz) Example 3 Yadandioside O? H HOOHCHz Fraction 21 (3.8 g) of chromatography using Column A in Example 2 was separated by the column, and the obtained fraction 16l62-215 (315 was further purified with Column C to obtain the title from fraction 39-41. 53 ■ of the compound were obtained.

Melting point: 183-188°C 3430, 1740.1680.1640.1065.
1050.1020UV spectrum (ethanol, λw
ax) :224nm (ε=15000), 2
54nm (ε=8000) mass spectrum (m/z
): 560 (M-162-60), 43B.

420.402, 392.141, 123.6ONMR
Spectrum (pyridine-dl, δ): 1.16 (38,
d, J=8.11Lz), 1.24(3H,t,J=
8.1Hz) 1.49(3)1. s), 1.51(3
H,s), 1.63(3H,s) 1.94(3H,s
), 2.50 (LH, d, J = 5.41 [z) 2.6
7 (IH, w), 2.78 (111, m), 3.8
6(LH,s).

5.05 (18, d, J = 8.1 Hz), 5.13 (
IH, d, J = 5.4Hz) 5.28 (LH, d, J =
8.1Hz), 6.10(IH,s).

7.27 (IH, s) Example 4 Yadandioside N-aglycone? 8 Yadandioside N25 obtained in Example 2 was dissolved in 15 ml of 3N sulfuric acid water-methanol (1:1) and heated under reflux for 3 hours. After concentration, extraction was performed with methylene chloride, and the resulting organic layer was washed with an aqueous sodium bicarbonate solution and saturated brine, and dried over anhydrous magnesium sulfate. The resulting residue was subjected to silica gel column chromatography (Wa
The reaction mixture was subjected to chromatography (Ko C-200) and eluted with methanol/chloroform (5:95) to obtain the title compound 12.

Melting point: 180-183℃ (α) z4: + 30'' (C-0,7,xl
)/-le) IR spectrum (KBr, cm-')
? 3460, 1?50.1685.1645.1050
．． 1010UV spectrum (ethanol, λn+ax)
:220nm (ε-16000), 267nn+
(ε=5000) High resolution mass spectrum (C!1H
j60□) Calculated value: 548.2257 Actual value: 548,2282 NMR spectrum (CDCf,, δ): 1.06 (6
H, d, J = 6.8 Hz) 1.11 (38, d, J = 7
．． 0) lz).

1.62 (3H, s), 2.14 (3H, d, J=1
．． 1Hz) 3.78 (3H, s), 4.29 (LH,
brs).

4.66 (ltl, brs), 4.70 (IH, d
, J=7.9Hz) 5.44 (1B, br), 5.
66 (IH, brs).

5.74 (LH, d, J=2.4Hz) Example 5 Yadandioside O-aglycone ♀H Yadandioside 072■ obtained in Example 3 was dissolved in 5 m (l) of water with 30■ The reaction solution was kept for 2 weeks. A small amount of ethanol was added to the reaction solution and heated. The coagulated enzyme was removed by filtration, and the solution was concentrated under reduced pressure.

The residue was subjected to column chromatography on silica gel (Wako C-200) and methanol/chloroform (
1:9) and chloroform/methanol/water (13
: 4 : When the lower layer of 1) was poured out, the title compound 13 Akebono and the recovered raw material (Yadandioside O) 3°3
I got ■.

Melting point: 138-143℃ [α] Temperature: +23° (C=0.6.ethanol) I
R spectrum (KBr, Cl11-'): 3450
, 1740.1640.1060 UV spectrum (ethanol, λmax): 222 nm (t = 15000
), 278 nm (g = 7000) mass spectrum (
m/z): 560 (M-60), 438,420
゜402.392,354,278.140.123.
111.6ONMR spectrum (CDCJ3.δ): 1
．． 13 (3H, t, J=7.3Hz), 1.40 (3
H, s) 1.57 (6H, s), 1.85 (3) 1.
d, J=2.2Hz) 2.03 (3H, 3), 2.5
9 (2H, Q, J = 7.2H2) 3.81 (3H9s)
+4.22 (IH, brs).

4.26 (LH,brd,J=4.3Hz).

4.73 (LH, d, J = 7.6Hz), 4.82 (
LH, brs).

5.78 (IH, s) 1 example: Honshu suction substitute investigation responsibility (7)? 11-
Effect on murine leukemia P388 The compounds of the present invention were tested for their effects on murine leukemia P388 using the compounds prepared in Examples 4 and 5.

The test was carried out by inoculating 10 b cells of murine leukemia P388 and administering the test compound intraperitoneally to a group of 6 BDFI mice (female) at the indicated dose for 5 consecutive days.

The considerations shown in the table below indicate the number of mice that died due to drug toxicity during the drug administration period (denominator) against the total number of mice (denominator), and the survival rate (ILS%) is calculated using the following formula: Calculated with.

Survival prolongation rate (ILS%) = [(Survival days of administration group)/(Survival days of control group))X100-100 (%) This control represents a drug-free group.

〔Effect of the invention〕

As shown in the table above, the yadandioside N-aglycone and the yadandioside O-aglycone of the present invention have excellent antitumor activity at low doses.

From this, these compounds are expected to develop in the future as new antitumor and antiviral drugs.

Claims (1)

[Claims] 1. General formula ▲ Numerical formula, chemical formula, table, etc. ▼ [In the formula, R is a 3,4-dimethyl-2-pentenoic acid residue or 3-ethyl-4-methyl-4-acyloxy- Represents a 2-pentenoic acid residue, and the group ▲ has mathematical formulas, chemical formulas, tables, etc. ▼ has ▲ mathematical formulas, chemical formulas, tables, etc. ▼, ▲ mathematical formulas, chemical formulas,
There are tables, etc. ▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (where Glc represents a glucose residue and Y represents a hydrogen atom or a glucose residue). However, when R is a 3,4-dimethyl-2-pentenoic acid residue, the group ▲ has a mathematical formula, chemical formula, table, etc. ▼ has a ▲ mathematical formula, chemical formula,
A new quassinoid compound that has a table, etc. ▼ or ▲ a mathematical formula, a chemical formula, a table, etc. ▼ (where Glc represents a group other than as defined above). 2, R is a 3,4-dimethyl-2-pentenoic acid residue, 3-
Claim 1, which is an ethyl-4-methyl-4-acetoxy-2-pentenoic acid residue or a 3-ethyl-4-methyl-4-propionyloxy-2-pentenoic acid residue.
Novel quassinoid compounds described in Section. 3. Brucea Javanica (L.)
Merr seeds) are extracted with alcohol, then the organic solvent fraction is collected and purified, or the general formula is characterized by a deglucose reaction after collection and purification ▲There are mathematical formulas, chemical formulas, tables, etc.▼ In the formula, R represents a 3,4-dimethyl-2-pentenoic acid residue or a 3-ethyl-4-methyl-4-acyloxy-2-pentenoic acid residue, and the group ▲ has mathematical formulas, chemical formulas, tables, etc. ▼ ▲There are mathematical formulas, chemical formulas, tables, etc.▼, ▲Mathematical formulas, chemical formulas,
There are tables, etc. ▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (where Glc represents a glucose residue and Y represents a hydrogen atom or a glucose residue). However, when R is a 3,4-dimethyl-2-pentenoic acid residue, the group ▲ has a mathematical formula, chemical formula, table, etc. ▼ has a ▲ mathematical formula, chemical formula,
▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ where Glc represents a group other than (as defined above)] A method for producing a novel quassinoid compound.