JPH07188001A - Benzaldehyde-based antiviral agent - Google Patents

Abstract

PURPOSE:To obtain a new antiviral agent containing a specified benzaldehyde which is an active component existing in a herb bennet extract. CONSTITUTION:An antiviral agent containing 3,4,5-trihydroxybenzaldehyde represented by the formula as the active component. This compound of the formula can be obtained by drying the whole plant of a herb bennet, boiling and extracting it in deionized water or distilled water, extracting the extracted solution with ethyl acetate, removing ethyl acetate to obtain the extract residue, treating it by the silica gel column chromatography method, eluting it with a chloroform-methanol mixture solvent (9:1), treating the eluted fraction by the silica gel chromatography method again, eluting it with an ethyl acetate- chloroform mixture solvent (1:4) and further purifying the eluted fraction by the reverse phase thin layer chromatography method using an acetonitrile-water mixture solvent (1:9).

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel antiviral agent.

[0002]

2. Description of the Related Art Although radish root has been used as a diuretic and a remedy for purulent disease since ancient times, the present inventors discovered for the first time that radish root had an antiviral activity, and it was found that the herb medicine consisting of radish root An application for a virus agent has been filed as Japanese Patent Application No. 5-123341.
At that stage, however, it was unclear which compound was effective because the radish plant contained a wide variety of compounds.

[0003]

Therefore, an object of the present invention is to identify the active ingredient present in the Radish root extract and to clarify the use of the compound.

[0004]

[Means for Solving the Problems] The present inventors have studied the antiviral activity of radish extract, and investigated the antiviral activity of each compound group whose chemical structure was determined by separating and purifying from the extract. . As a result, the present inventors have confirmed that 3,4,5-trihydroquinbenzaldehyde is present in Radish and have antiviral activity, and completed the present invention.

Radish (Geum Japanic)
500 g of dried whole um Thunb) is boiled and extracted with 5.4 liters of deionized water or distilled water, and the extract is extracted with 6 liters of ethyl acetate. The ethyl acetate was removed to obtain 5.3 g of an extraction residue. 30 g of the extraction residue was subjected to silica gel column chromatography, and 14.4 g of the fraction eluted with chloroform: methanol (9: 1) was subjected to silica gel column chromatography again, and the fraction eluted with ethyl acetate: chloroform (1: 4) was reversed. Further purification by thin-layer chromatography with acetonitrile: water (1: 9)
4,5-Trihydroxybenzaldehyde was obtained. The above fractionation method is as follows.

[0006]

[Table 1]

Infrared spectrum, melting point, NM of this product
Each R was identical to that of 3,4,5-trihydroxybenzaldehyde obtained separately, and the structure was confirmed by this. Figure 1 shows the infrared spectrum of 3,4,5-trihydroxybenzaldehyde.

In order to examine the antiviral activity, herpes simplex virus type I 7401H strain, poliovirus (vaccine strain, Sabin strain) and measles virus (vaccine strain, Tanabe strain) were used as viruses. Virus is Vero
Infect cells (African green colander cells), and after a certain period of time, freeze and thaw the infected cells three times and then centrifuge (3000
(rpm, 15 minutes) to give a supernatant virus solution, which was used for the virus test. The virus solution was stored at -80 ° C. All cells were cultured in a carbon dioxide incubator at 37 ° C.

Cytopathic effect [Cytophathic
The plaque formation test for quantifying virus using the effect; (CPE)] as an index was conducted as follows. Vero cells monolayer-cultured on a 60 mm plastic petri dish were inoculated with the diluted virus solution (0.2 ml) prepared by adjusting the virus solution to a concentration of 100 plaques / 0.2 ml, and the cells were allowed to adsorb the virus for 1 hour at 37 ° C. It was After adsorption, a solution of radish root extract or 3,4,5-trihydroxybenzaldehyde having a concentration shown in Table 2 and 2 wt% fetal calf serum-containing MEM medium (5 ml) containing 0.8 wt% methylcellulose was used to "adsorb the virus. Single layer Vero
The cells were overlaid and an attempt was made to form plaques at 37 ° C for 2-5 days. To calculate the number of plaques, the monolayer cells were fixed with formalin and then stained with 0.03% methylene blue solution. The antiviral activity was examined a plurality of times as described above using the drug extract solution having the same concentration, and it was confirmed that the assay results were within the plaque confirmation judgment error.

[0010]

[Table 2] (Note) The drug I is 3,4,5-trihydroxybenzaldehyde, and the drug II is a hot water extract of Radish root.

From Table 2, it became clear that 3,4,5-trihydroxybenzaldehyde exhibits antiviral activity. At this time, no cytotoxicity due to 3,4,5-trihydroxybenzaldehyde was observed. This data is for the case where 3,4,5-trihydroxybenzaldehyde was used alone as an antiviral agent.

[0012]

【effect】

(1) It was confirmed that 3,4,5-trihydroxybenzaldehyde was a new antiviral agent. (2) Radish root extract containing 3,4,5-trihydroxybenzaldehyde is also useful as an antiviral agent. (3) The compound does not show cytotoxicity.

[Brief description of drawings]

FIG. 1 shows an infrared absorption spectrum of 3,4,5-trihydroxybenzaldehyde.

Front page continuation (72) Inventor Kimayasu Shiraki 2556-4 2-202 (72) Inventor Shigetoshi Kadota 2556-4 2-401 Gosufuku-cho, Toyama City, Toyama Prefecture Inventor Kurokawa Masahiko 2-2 2-101 Minamitaikoyama, Kosugi-cho, Imizu-gun, Toyama Prefecture

Claims (1)

[Claims] Claims: An antiviral agent comprising 3,4,5-trihydroxybenzaldehyde represented by the formula (3) as an active ingredient.