JPH0196120A - Anti-inflammation remedying agent - Google Patents

Abstract

PURPOSE:To obtain an anti-inflammation remedying agent having high safety, by using a sesquiterpenoid compound as an active component. CONSTITUTION:The objective agent contains the sesquiterpenoid compound of formula (named as dehydrocurdione) as an active component. The compound of formula can be separated from an organic solvent extract of GAJUTSU (rhizome of Curcuma zedoaria, a herb drug). Concretely, the separation is carried out by extracting GAJUTSU with a nonpolar organic solvent such as n-hexane, ether, benzene, ethyl acetate, etc., and fractionating the extract by column chromatography, etc. The fraction of the compound of formula can be obtained after distilling out the solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to an anti-inflammatory therapeutic agent containing a sesquiterpenoid compound (named devidrocludione) represented by the following structural formula (I) as a main component.

BACKGROUND OF THE INVENTION Sesquiterpenoid compounds represented by the above structural formula
) 987 (1972) Il, 1Iikino et al., it was isolated from the organic solvent extract of zedoariae Rhizoma (scientific name: Zedoariae Rhizoma, Lt.), and is a known substance. It is.

By the way, kajutsu is a herbal medicine that has been known since ancient times, and its ingredients include essential oils such as cineole, sesquiterpene alcohol, and camphene, other fatty oils, starches, sticky substances, and rubbers, which have aromatic and healthful properties. It is used as a stomach medicine.

For example, Japanese Patent Application No. 60-066534 describes that gajutsu has an anti-ulcer effect, and proposals have also been made to use it as an anti-ulcer therapeutic agent. The above-mentioned sesquiterpenoid compounds were isolated from among the components of this gajutsu as substances with anti-ulcer effects.

Problems to be Solved by the Invention However, it is not known that the compound represented by the above structural formula (1) has an anti-inflammatory effect, and it is difficult to discover this effect and develop an anti-inflammatory therapeutic agent. It was long awaited.

Means for Solving the Problems In order to solve the above problems, the present invention provides an anti-inflammatory therapeutic agent containing a sesquiterpenoid compound represented by the following structural formula (1) as an active ingredient.

In order to produce the compound of the above structural formula (1), JP-A No. 57
This is carried out by isolating it from Gajutsu as described in Japanese Patent No. 163373. To perform this isolation, first, extraction is performed with a nonpolar organic solvent such as n-hexane, ether, penzole, and ethyl acetate, and the extracted extract is fractionated using column chromatography or the like. The solvent of both of the fractionated compounds of the structural formula (1) is distilled off under reduced pressure to obtain colorless plate-like crystals of the compound of the structural formula (1).

The physical properties of the compound of structural formula-(1) thus obtained include melting point, specific optical rotation, elemental analysis values, as described in detail in the above Chem, Pharm, Bull.
It can be identified by molecular weight, ultraviolet absorption spectrum, infrared absorption spectrum, nuclear magnetic resonance spectrum, etc.

Next, the pharmacological effects, effective amount, formulation method, etc. of the sesquiterpenoid compound of structural formula (I) will be explained.

(a) Pharmacological effect ■ Creation of experimental model Using Wistar male rats (female), the compound of the above structural formula was administered at 80 mg/Kg and 200 mg/Kg, respectively.
Thirty minutes later, 1% λ-Carrageenin dissolved in physiological saline was administered subcutaneously to the footpad of the right hind leg (0.1 ta It) as an inflammatory agent.
/head to create an experimental inflammation model.

■ Standard volumetric method Place a 30ml beaker containing 1% Tween 80 aqueous solution on a 1mg electronic dish balance, adjust the display scale to 0, and then press the foot! One part was gradually immersed into the liquid up to a certain point without contacting the container. The weight displayed is the weight of the liquid equivalent to the volume of the foot in the liquid. Therefore, the paw volume was expressed in weight units (g) equivalent to the volume of water.

(2) Measurement of response threshold for pain sensation Using a Randall-Selitto pressurizer, the response threshold value for pain at the inflamed site and foot stepping induced by administration on the inflamed side was measured and expressed in weight 1 (X10 g).

(2) Evaluation of anti-inflammatory effect As an index of anti-inflammatory effect, the pain suppression rate (al edema suppression rate and peak) was calculated as follows, and the presence or absence of a significant difference between it and the control was examined using a 5student test.

Indomethacin (Indoa+ethacin)? ,
The degree of effect was examined using 2 mg/Kg as a control.

(Left space on this page) Weight change in anti-inflammatory effect test Edema rate and pain in anti-inflammatory effect test! r! fIIl
Rate umbrella: P<0.05, **: P(0,01, m: P<0
．． 005 Near 7)
As a result, the sesquiterpenoid compound represented by the above structural formula (I) was found to have a strong anti-inflammatory effect against carrageenan edema.

As described in detail, according to the present invention, the above structural formula (1)
It has been discovered that the compound represented by the above has an anti-inflammatory effect, and as a result, it is possible to provide an anti-inflammatory therapeutic agent containing this compound as an active ingredient. If this anti-inflammatory treatment is made from crude drugs, its toxicity has already been proven to some extent.
It is possible to provide pharmaceuticals with high reliability in terms of safety.

Example A detailed explanation will be given below based on examples.

As shown in the figure, gajutsu from Yakushima (dried thin pieces, 10.
8Kg) in methanol (206) for 7 hours (water bath,
50°C) Extract. The same operation was repeated 9 times, the methanol extracts were combined, and the solvent was distilled off under reduced pressure to obtain methanol extract (abbreviated as YK-1, yield: 1.12 kg, yield from crude drug: 10.4%, below) (similar) was obtained.

Dissolve YK-1 (200 g) in methanol (600+Jt) and extract three times with hexane (3Il). remaining Y
A portion (800 g) of K-1 was also partitioned and extracted with methanol-hexane. The methanol layers were combined, the solvent was distilled off under reduced pressure, and the methanol layer extract (YK-2, 680 g,
7.1%). The hexane layers were combined, the solvent was distilled off under reduced pressure, and the hexane layer extract (YK-3, 310 g,
3.2%).

YK-3 (100g) was subjected to silica gel column chromatography [silica gel 3.0Kg (manufactured by MERCK, K
ieselge160), elution solvent; (1) hexane-
Ethyl acetate system (hexane: ethyl acetate = 50:1.20
:1.15:1.10:1.8:1 in the order of 20 liters each, (21 chloroform-methanol system (chloroform:methanol = 10:1.0:100 in the order of 8!)) (YK-4 to 8).

That is, the solvent in the eluate from the elution solvent of hexane:ethyl acetate = 20:1 was distilled off under reduced pressure, and the 4th minute @YK-
4C23,'3a, 0.75%) was obtained. Similarly, hexane:ethyl acetate-15:1. The 5th minute WJ (YK-5
, 14.3g, 0.46%), 6th minute WJ (YK-6,
1), 8g, 0.38%), 7th fraction (YK-7,
6.5g, 0.21%), 8th fraction from the chloroform:methanol=10:1 eluate (YK-8, 22.9g
, 0.74%) were obtained.

The solvent of YK-5 was distilled off under reduced pressure to obtain the sesquiterpenoid compound of the above structural formula (1) in the form of colorless plate-like crystals.

The physical properties of the obtained compound are as follows.

(1) Melting point: 37-39°C (hexane) (2)
Specific optical rotation: [α) D +67.9° (C = 0.51, chloroform) (3) Elemental analysis value: C,, +1□20□ (4
) Infrared absorption spectrum': "n"j1710 (cyclodecanone), 1
687 (cyclodecanone) (5) Ultraviolet absorption spectrum λ'5::" (fogg): 2241nf (3,63
),254inf(3,46)305inf(2
,56) (61'll-NMR(100, MIIz CC14,
δ): 0.99 (3H, d, J=6), 1.60 (3H
, d, J=15) 1.72 (3H, S) , 1.74 (
31), S) 2.94 (IH, d, J=1)), 3.
10(1)1. d, J=18) 3.14(III, d
, J = 1)) , 3.14 (Ill, d, J = 1
8) 5.08 (1) 1, dd, J = 157.5) (
7)') l-NMR (100 MIIz, da-benzene, δ): 0.75 (31, d, J=(i), 1.
48 (3H, S) 1.60 (3H, S), 1.66 (3
8, S) 2.90 (1) 1. d, J = 1)),
2.97(2H,S)3.1)(IH,d,J =1
)) , 5.00 (IH, dd, J = 7.575.

(8) It has no odor and is colorless plate-like crystals.

(9) Slightly soluble in methanol and ethanol, easily soluble in hexane and petroleum ether, and insoluble in water.

The structure and carbon positions determined as above are as follows.

When the pharmacological effects of each fraction prepared in this way were investigated according to the above-mentioned pharmacological effect test method, each fraction was found to have an 80.2
Anti-inflammatory effects were observed when administered orally at 00 mg/Kg. Among these, the YK-5 fraction was particularly strong. Therefore, when 80 mg/Kg of the compound of structural formula (1) obtained from YK-5 was orally administered, a stronger anti-inflammatory effect was observed as described above.

Examples of formulations are shown below.

Formulation example 1 (100 tablets) Dividerocurdione 8.0g potato starch 1). 5 g Magnesium stearate 0.5 g Total amount 20
．． 0 g Divide Rocludione 8 g and potato starch 1). 5 g was mixed well, water was added to the mixture, the mixture was kneaded, and the mixture was granulated using a screen having a mesh size of 1■HXl+u to form granules. After drying, the particles are sized using a No. 16 mesh (B, S) sieve. Next, 0.5 g of magnesium stearate was mixed with the granules, and one tablet of 20
It was made into a 0 mg tablet. In addition, it is also possible to apply a usual easily soluble film coating to this tablet, if necessary.

Formulation example 2 (100 tablets) Dividerocludione 8.0g Lactose (20
0 mesh) 70.0 g potato starch 12.0 g gelatin
10.0 g total amount 100
．． 8 g of 0 g divided locludione is mixed with an equal amount of 8 g of potato starch and the mixture is ground. Add 4g of potato starch and 70g of lactose to this and mix. Separately, add 10g of gelatin to 100ml of purified water.
After cooling, ethanol Loom It is added to this solution while stirring to obtain a gelatin solution as a binder.

This gelatin solution is added to the above-mentioned mixture of dividrocludione and the like, and granulated and sized according to a conventional method.

Although the above description has been made for internal use, it can also be used as an external preparation.

[Brief explanation of the drawing]

The figure shows an example of the method for producing the substance represented by the above structural formula (1) used in this anti-inflammatory therapeutic agent, in which the substance is isolated by active fractionation of Yakushima Gajutsu. Show the process. October 7, 1988 Jin\Idrokuru 32 Reprimand Procedural Amendment (Voluntary) January 20, 1988 Patent Office Commissioner Kunio Ogawa 1, Indication of Case 1988 Patent Application No. 25171) 2. Name of the invention Anti-inflammatory therapeutic agent 3. Relationship with the case of the person making the amendment Patent applicant 3-5-21 Shakujiidai, Nerima-ku, Tokyo Representative of Long-established Keimeido Co., Ltd. Ken Shiba Ken Satoru 4, Agent ■ 105 5. Date of amendment order Voluntary 6. Number of inventions to be increased by amendment None 7. Subject of amendment "Column for detailed explanation of the invention in the specification" 8. Contents of amendment (1) Specification, page 2, No. 16 In the line, "Japanese Patent Application No. 1988-066534" is corrected to "Specification of Japanese Patent Application No. 60-066534." (2) On page 2, line 20 of the specification, the phrase "sesquiterpenoid compound" is corrected to "sesquiterpenoid compound." (3) Add the following sentence between the third and fourth lines of page 9 of the specification. Next, we will explain the results of the acute toxicity test. SLC: Using 5-week-old male and female ICR mice, they were preliminarily housed in a test environment for 8 to 10 days, and it was confirmed that there were no abnormalities. From evening until 3 hours after administration (
The animals were fasted for 20 hours in total and given only water for that amount. After each concentration solution was cleaved so that the administration volume was 0.2 ml per 10 g of body weight, it was forcibly administered directly into the stomach using a metal stomach tube. Five male and female animals were used for each dose. Under the above conditions, the LD5Q value was determined by the Litchfield-Wilcoxon method based on the cumulative mortality rate on day 14 (SOP: ETN-14). The results were as shown in the table below. As shown in the table above, the effective dose of the anti-inflammatory therapeutic agent is very small compared to the LD5Q value, so it is considered that there is no problem with acute toxicity. "that's all

Claims (1)

[Claims] (1) An anti-inflammatory therapeutic agent containing a sesquiterpenoid compound represented by the following structural formula (I) as an active ingredient. ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I)