JPH0523243B2 - - Google Patents

Description

[Detailed description of the invention]

Industrial Application Field The present invention is based on the following structural formula () The present invention relates to an anti-inflammatory therapeutic agent containing a sesquiterpenoid compound (named dehydrocurdione) as a main component. Prior Art The sesquiterpenoid compound represented by the above structural formula () is, for example, Chem.Pharm.Bull 20(5)
987 (1972) H. Hikino et al., it was isolated from the organic solvent extract of Zedoariae Rhizoma (scientific name: Zedoariae Rhizoma, ginger family) and is a known substance. By the way, Gajiyutsu 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, starch, stickiness, and rubber, and are used as aromatic stomachic medicines. It is used as. For example, Japanese Patent Application No. 60-066534
No. 227575) describes that gajiyutsu has an anti-ulcer effect, and there is also a proposal to use it as an anti-ulcer therapeutic agent. The above-mentioned sesquiterpenoid compounds were isolated from among the components of this gajiyutsu as substances with anti-ulcer effects. Problems to be Solved by the Invention However, it is not known at all that the compound represented by the above structural formula () has an anti-inflammatory effect, and it is long-awaited to discover this effect and develop an anti-inflammatory therapeutic agent. It had been. Means for Solving the Problems In order to solve the above problems, the present invention provides the following structural formula () The present invention provides an anti-inflammatory therapeutic agent containing a sesquiterpenoid compound as an active ingredient. The compound of the above-mentioned structural formula () can be produced by isolating the compound from gajiyutsu as described in JP-A-57-163373. To perform this isolation, first use n-hexane, ether,
Extraction is performed with nonpolar organic solvents such as benzene and ethyl acetate, and the extracted extract is fractionated using column chromatography or the like. The solvent of the fractionated compound of the above structural formula () is distilled off under reduced pressure to obtain the compound of the above structural formula (2) in the form of colorless plate-like crystals. The physical properties of the compound of the above structural formula () thus obtained include melting point, specific rotation, elemental analysis values, molecular weight, ultraviolet absorption spectrum, as described in detail in the above Chem.Pharm.Bull. It can be identified by infrared absorption spectrum, nuclear magnetic resonance spectrum, etc. Next, the pharmacological effects, effective amount, formulation method, etc. of the sesquiterpenoid compound of the above structural formula () will be explained. (a) Pharmacological effect Creation of experimental model Using male Wistar rats (female), 80 mg/Kg and 200 mg/Kg of the compounds with the above structural formula were orally administered by force, and 30 minutes later, the compounds were dissolved in physiological saline. An experimental inflammation model was created by subcutaneously administering 1% λ-Carrageenin as an inflammatory agent to the right hind leg footpad at a rate of 0.1 ml/head. Footpad volume measurement method Place a 30ml beaker containing 1% Tween 80 aqueous solution on an electronic dish balance with a sensitivity of 1mg, and after adjusting the display scale to 0, gradually measure the volume of the footpad up to a certain point without touching the container. Immersed in liquid. The weight displayed is the weight of the liquid equivalent to the volume of the foot in the liquid.
Therefore, the foot volume is the weight unit (g) of the same volume as water.
It was expressed as Method for Measuring Response Threshold for Pain Sensation Using a Randall-Selitto pressure device, the response threshold for pain at the footpad inflammation site induced by administration of the inflammatory agent was measured and expressed in weight (×10 g). Evaluation of anti-inflammatory effect As an index of anti-inflammatory effect, (a) edema suppression rate and (b) pain suppression rate were calculated as follows, and the presence or absence of a significant difference between them and the control was examined by Student's t test. (a) Edema rate = Edema footpad volume - Normal footpad volume / Normal footpad volume × 100 (b) Pain suppression rate = Edema pain threshold - Normal pain threshold / Normal pain threshold × 100 Positive control Indomethacin (1.2 mg/Kg), which has edema-suppressing and analgesic effects, was used as a control to examine the degree of effectiveness.

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[Table] As a result, the sesquiterpenoid compound represented by the above structural formula () was found to have a strong anti-inflammatory effect against carrageenan edema. Next, the results of the acute toxicity test will be explained. Using 5-week-old male and female SLC:ICR mice, they were preliminarily bred for 8 to 10 days in a test environment, and it was confirmed that there were no abnormalities. The animals were fasted from the evening of the day before drug administration until 3 hours after administration (20 hours in total), during which time they were given only water. After each concentration solution was prepared 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 LD ₅₀ 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.

[Table] As shown in the table above, the effective dose of the anti-inflammatory therapeutic agent is very small compared to the LD ₅₀ value, so there is no problem with acute toxicity. Effects of the Invention As explained above, according to the present invention, it has been found that the compound represented by the above structural formula () has an anti-inflammatory effect, and thereby it is possible to provide an anti-inflammatory therapeutic agent containing this compound as an active ingredient. . If this anti-inflammatory therapeutic agent is made from crude drugs, its toxicity has already been proven to some extent, so it is possible to provide a drug with high reliability in terms of safety. EXAMPLE A detailed description will be given below based on an example. As shown in the figure, Yakushima gajiyutsu (dry flakes, 10.8 kg) was extracted by digestion (water bath, 50°C) with methanol (20) for 7 hours. The same operation was repeated 9 times, the methanol extracts were combined, and the solvent was distilled off under reduced pressure to obtain the methanol extract (abbreviated as YK-1, yield 1.12 kg, yield 10.4% from crude drugs, the same applies hereinafter). Obtained. Dissolve YK-1 (200 g) in methanol (600 ml) and extract three times with hexane (3). Remaining
A portion of YK-1 (800 g) is 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, 680g, 7.1%) was obtained. The hexane layers were combined, the solvent was distilled off under reduced pressure, and the hexane layer extract (YK
-3, 310g, 3.2%) was obtained. YK-3 (100g) was subjected to silica gel column chromatography [silica gel 3.0Kg (MERCK, Kieselgel 60), elution solvent: (1) hexane-ethyl acetate system (hexane:ethyl acetate = 50:1, 20:1, 15 :1,10:1,8:
(20 each in the order of 1), (2) Chloroform-methanol system (chloroform:methanol = 10:1, 0:
Fractionation using each 8] in the order of 100 (YK-4 to 8)
do. That is, the solvent of the eluate from the elution solvent of hexane:ethyl acetate = 20:1 was distilled off under reduced pressure to obtain the fourth fraction YK-4 (23.3 g, 0.75%). Similarly, hexane:ethyl acetate = 15:1, 10:1, 8:1
The fifth fraction (YK-5, 14.3 g, 0.46%) and the sixth fraction (YK-6,
11.8g, 0.38%), 7th fraction (YK-7, 6.5g, 0.21
%) and the eighth fraction (YK-8, 22.9 g, 0.74%) were obtained from the chloroform:methanol=10:1 eluate. 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℃ (hexane) (2) Specific optical rotation: [α] _D +67.9° (C = 0.51, chloroform) (3) Elemental analysis value: C ₁₅ H ₂₂ O ₂ (4) Infrared absorption spectrum ν ^CCl _Max 4cm ^-1 : 1710 (cyclodecanone), 1687 (cyclodecanone) (5) Ultraviolet absorption spectrum λ ^MeOH _Max (logε): 224inf (3.63), 254inf (3.46)
),
305inf (2.56) (6) ¹ H−NMR (100MHzCCl ₄ , δ): 0.99 (3H,
d, J=6), 1.60 (3H, d, J=15) 1.72
(3H, S), 1.74 (3H, S), 2.94 (1H, d,
J=11), 3.10 (1H, d, J=18), 3.14 (1H,
d, J=11), 3.14 (1H, d, J=18), 5.08
(1H, dd, J=757.5) (7) ^1H -NMR (100MHz, _d6 -benzene, δ):
0.75 (3H, d, J=6), 1.48 (3H, S),
1.60 (3H, S), 1.66 (3H, S), 2.90 (1H,
d, J=11), 2.97 (2H, S), 3.11 (1H, d,
J = 11), 5.00 (1H, dd, J = 7.575. (8) It has no odor and is a colorless plate-like crystal. (9) Slightly soluble in methanol and ethanol, easily soluble in hexane and petroleum ether, It is insoluble in water. The structure and carbon positions determined above are as follows. When the pharmacological effects of each of the fractions thus prepared were investigated according to the pharmacological effect test method described above, anti-inflammatory effects were observed when 80 and 200 mg/Kg of each fraction was orally administered. Among these, the YK-5 fraction was particularly strong. Therefore, when 80 mg/Kg of the compound of the above structural formula () 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) Dehydrocurdione 8.0g Potato starch 11.5g Magnesium stearate 0.5g Total amount 20.0g Mix 8g of dehydrocurdione and 11.5g potato starch well, and add water to the mixture. In addition, the kneaded mixture is granulated using a screen having a mesh size of 1 mm x 1 mm to form granules. After drying, sieve with a No. 16 mesh (BS) sieve. Next, 0.5 g of magnesium stearate was mixed with the granules, and each tablet was made into a 200 mg tablet using a tablet machine. In addition, it is also possible to coat this tablet with a usual easily soluble film coating, if necessary. Formulation example 2 (100 tablets) Dehydrocurdione 8.0g Lactose (200 mesh) 70.0g Potato starch 12.0g Gelatin 10.0g Total amount 100.0g Dehydrocurdione 8g was mixed with an equal amount of potato starch 8g. Grind the mixture.
Add 4g of potato starch and 70g of lactose to this and mix. Separately, add 10g of gelatin to purified water.
After cooling, add 100 ml of ethanol to this solution while stirring to obtain a gelatin solution as a binder. This gelatin solution is added to the above-mentioned mixture of dehydrocurdione, etc., 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 a method for producing the substance represented by the above structural formula () used in this anti-inflammatory therapeutic agent.
The process of isolating this substance by active fractionation is shown.

Claims (1)

[Claims] 1. An anti-inflammatory therapeutic agent containing a sesquiterpenoid compound represented by the following structural formula () as an active ingredient.