JPS5953477A - Coumarin derivative - Google Patents

Abstract

NEW MATERIAL:A coumarin derivative shown by the formula I (R is group shown by the formula II or formula III). USE:Useful as a drug having antiallergic actions. PROCESS:Cnidii monnieri fructus is extracted conventionally with an extraction solvent such as an alcohol to give an extracted essence, this extracted essence is fractionated by column chromatography in combination with thin layer chromatography to give a compound shown by the formula I . In a concrete preparation example, Cnidii monnieri fructus of China growth on the market is coarsely powdered, soaked in 50wt% warm aqueous solution of methanol to give a methanol essence. The essence is shaked with water-n-butanol, components transferred to the n-butanol layer are subjected to column chromatography using silica gel as an adsorbent and benzene: acetone of (50:1) 5:1 as an eluting solvent, and fractionated by thin layer chromatography using benzene or acetone as a developing solvent. The prepared fraction II is fractionated by thin layer chromatography to give a compound shown by the formula I .

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel marine derivative contained in japonica.

Snake beetroot is a herbal medicine that has long been listed in the Shennong Bencaojing, and has been used as an astringent anti-inflammatory2, but its active ingredients are unknown, and its effectiveness itself has not been evaluated pharmacologically. Not proven.

The present inventor conducted numerous experiments with the aim of extracting components with anti-allergic effects from natural products.
We found that the methanol extract of Jatoko has a mild but key anti-allergic effect, and as a result of further research, we discovered the following new marine derivatives as ingredients with anti-allergic effects: CH, -CH = CCCJI, )2 The present invention relates to a compound of formula (1) and a compound of formula (1), which have relatively similar structures among the compounds of the above formula.

In order to obtain the compound of formula (1) and the compound of formula (1) from japonica, the japonica is subjected to a conventional extraction process using alcohol as an extraction solvent to obtain an extract, and then this extracted extract is diluted. Fractionation is performed by column chromatography in combination with layer chromatography to obtain a fraction containing mainly coumarin derivatives.

In addition to the compound of the formula (1) and the compound of the formula (1), which are the compounds of the present invention, the fraction containing mainly l-coumarin derivatives includes the above-mentioned compound of the formula (1) and these novel compounds. Although a known compound of the formula (ff) structurally similar to the compound is present, the fraction mainly containing the coumarin derivative was separated by column chromatography rather than by thin layer chromatography. This can be done by separating the compound of the present invention - the compound of formula (member) and the compound of formula (1), the compound of formula (I), and the compound of formula (II').

The compound of formula (member) and the compound of formula (1) in Jaboko are:
It is easier to obtain these as a mixture than to obtain them completely separated, and when the compound of the present invention is used as an active ingredient of an antiallergic agent, the compound of formula (1) and ( 1) The use of a mixture of compounds of the formula does not cause any pharmacological disadvantage.

Therefore, when the compound of the present invention is obtained using Jatoko as a raw material and is used together with the active ingredient of an antiallergic agent,
Rather, it is better to obtain the compound of formula (1) and the compound of formula (1) as a mixture without separation, and use this mixture as an active ingredient of an antiallergic agent.

In this case, the dosage in terms of active ingredient may be set in the same way for the former anti-allergic agent using a single compound and the latter anti-allergic agent using a mixture.

Hereinafter, the present invention will be explained in detail with reference to production examples, pharmacological test examples, toxicity test examples, etc.

Production example 1) After coarsely powdering a commercially available china beetroot, it was
% methanol aqueous solution three times to obtain a methanol extract.

2) The methanol extract obtained above was shaken with water-n-butanol. The components transferred to the n-butanol layer were mixed with benzene/acetone 50% using silica gel as an adsorbent.
1 → 5:1 as an eluent, and thin layer chromatography using benzene:acetone as an 8-di opening solvent. A fraction (Ily yield: 4.5%), a fraction (Ily yield: 522%), a fraction (yield: 4.0%), and a fraction - (yield: 39.5%) were obtained.

5) Fraction 125.98F obtained above was mixed with n-hexane:ethyl acetate/L/4:1→2: using silica gel as an adsorbent.
1 as an elution solvent.
The figure shows, in order, a compound of formula (ff), a compound of formula (1), and (
Figures 5 to 8 show the NMR spectra of the compound of formula (Rin) and the compound of formula (1).
PEAK iFigure 5 M/B 2440 NT 234.
4, Fig. 6 M/m 202.0 Engineering NT 715.3, Fig. 7 M/'lRi 19Q, QINT 204.5, Fig. 8 M/l! ! 43.0 engineering NT 422.7).

Pharmacological Test Example 1) Effect on Delayed Dermatitis An experiment was conducted using picryl chloride as an allergic reaction inducer using 1 group of male BDF mice weighing around 21 mm. That is, a 7% picrylic chloride ethanol solution was applied to the abdomen of a mouse with prickly hair to perform the first sensitization, and immediately thereafter, the test drug dissolved in distilled water was orally administered twice a day for 10 days. .

The dosage of the test drug was as follows.

Example 2) Fraction 1; Fraction 1 of 1100rnΔ;
250mfA fraction ■; 100ml AQ min iJj y ; 250m1AQ To determine the medicinal efficacy, apply a 1% chlorinated picryl olive oil solution to both ears of a mouse, and immediately before the reaction and after the reaction 12.1B, 24
After 36 hours, the degree of antiflag inflammation caused by applying 1% picryl chloride was measured using a dial six gauge (PEAC).
The measurement was carried out using an OK-G model (Amagasaki Seisakusho).

The results are as shown in Figure 9, and are the edema curve of fraction 1 in the diagram of production example 2) and the edema curve of fraction 1 (edema curve of 2 in the diagram).
Particularly effective was seen.

Regarding fraction 1 of Production Example 2), the same test as above was conducted using a dose of 550 ml as a test drug, and the results shown in Figure 11) were obtained.

Furthermore, for the compound of Production Example 3), the same test as above was conducted using a dose of 100rnlA9 as a test drug. For comparison, for fraction 1 of Production Example 2), the test was conducted again at a dose of 1 oorrnAq.

The results are as shown in Figure 11, and are shown in Production Example 2)
Minute l! ! ! Il (edema curve 2 in the figure) and Production Example 3
), the compound of formula (1) (edema curve 6 in the figure) was particularly effective, and i! I! ! The compound of formula (1) in Example 3) (
A clear effect, although weaker than the former, was observed for the compound of formula (1) (edema curve 7 in the figure) and edema curve 1 (curve 8 in the figure). In Production Example 3), the compound of formula (IV) (edema curve 5 in the figure) had almost no effect.

2) Effect on PCA reaction Method for preparing antiserum Dissolve bovine 111 serum albumin (BSA) in a ratio of 2 parts in physiological saline, and add enough Freund's Combrete Azierband in a ratio of 1:1. Emulsify and mix 1. A male guinea pig weighing around 300 g was lightly anesthetized with ether, and the milk 1 solution was applied to a total of 4 sites on the front and back legs at 0.11 μm.
Injected subcutaneously. After this injection, the antigen BSA was injected into the back for the 300th time in two places with 0.1 ml in the same manner as described above, and the second sensitization was carried out. On the 14th day after the second cropping, whole blood was collected to obtain serum, which was used as anti-BSA serum in the following experiments.

Experiment method: Hair pricked with electric clippers on the day before the experiment 3
Immediately after oral administration of the test drug to a group of 5 male Hartley guinea pigs weighing approximately 0.00 g, the diluted antiserum was injected subcutaneously into the back at 13 locations of 0.1 m. 3 hours after injection, the antigen BSA O, 2 and physiological saline solution O, 5
Shoulder l and Epansub 1v-5% physiological saline solution O,S
Mix it with crocodile t and inject it through the front leg vein. 30 minutes after intravenous injection
Bleed to death for a minute and peel off the skin on the back. The degree of anti-allergic effect was determined based on the area of pigment spots induced on the skin using a planimeter. The results are shown in Table 1.

It is clear from Table 1 that the QI is weak compared to prednisolone, which is a control drug, but it is still as good as 11 in production example 2).
111 was clearly shown to have a PCA reaction inhibitory effect. Still, 9;! ! We also have the tsukunor extract in example 1).
It was f.

Acute Maternal Test LD5o was determined from lethal grains 3 days after oral administration of the test drug using 15 lineal male mice weighing 1B-20f per group. The results were as shown in Table 2.

From the results shown in Table 2, it can be seen that the compounds of the formula (1) and the compound of the formula (■), which are the compounds of the present invention, can be used for 400 to 600 m/d, 1
It is estimated to be effective in treating allergies in adults when taken two to three times a day. In particular, the compound of the present invention is thought to exhibit remarkable antiallergic effects when formulated with a compound of formula (2).
1) An anti-allergic agent that is effective for treating allergies in adults when taken in smaller doses than the anti-allergic agent consisting of the compound of formula 200-300 mt/dose, 2-3 times a day. It is estimated that this can be achieved.

Below, further examples of formulations in which the compound of formula (1) is used are given.

A compound of formula (1) is thoroughly mixed with Paleisi starch,
Add water and mix. The mixture is granulated using a screen having a mesh size of 1JIJl x 1MM to form granules. After drying &16
Sort the particles using a Metsushi (B, $.) sieve. Next, the granules were mixed with magnesium stearate and 1T30
There are 0 tablets and 1~.

[Brief explanation of drawings]

Figures 1 to 4 show the NMR spectra of the compound of the formula (IV), the compound of the formula (■), the compound of the formula (sulfur), and the compound of the plaster formula, and Figures 5 to 8 show the MA8B spectra. show. Figures 9 to 11 are graphs showing the test results for delayed-type relapsing inflammation (due to picryl chloride). In the figures, 10 indicates the edema curve of the blind group, and 10 indicates the edema curve of the control group. means. Also, the sign of the ability has the following meaning. Meaning of symbols in FIG. 9 1; Fraction 1 of 2) of Production Example (10o#, 4Lj administration)
Edema curve of 2N I fraction 1 (250 kgq administration) 13N I fraction I (9 in 100 mg/kg)
Administration) 14; l Fraction IV (2501q administration) #Meaning of the symbols in Figure 1+1 2; Edema curve of fraction l (350ml/b administration) of Production Example 2) Meaning of the symbols in Figure 11 Patent applicant Research Institute for Production Development

Claims (1)

[Claims] ■ General formula Marine derivative.