JP2000191520A - Microbicide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a microbicide capable of being relatively stably and continuously supplied and scarcely having side effects by confirming the effectiveness of a chemical substance which is an industrial product having a desirable microbicidal activity. SOLUTION: This microbicide contains one or more microbicidally active compounds selected from guaiacol, lignin sulfonic acid, 2,6-dimethoxyphenol, 3,5-dimethoxyphenol, lignosulfonic acid sodium salt, lignosulfonic acid sodium salt acetate, lignin organosorb propionate, 4-benzyloxyguaiacylglycerol-β-guaiacyl ether, syringic aldehyde and veratryl alcohol. Since the microbicide is an industrial product stable in the quantity and the property differently from natural substances, the antiviral property and antibacterial property of the microbicide can safely be achieved at a low cost.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an antimicrobial agent, and more specifically, it comprises guaiacol and lignin, and chemicals related to them as active ingredients, and comprises AIDS virus, influenza virus, etc. The present invention relates to an antimicrobial agent having an inhibitory effect on viruses and various bacteria.

[0002]

BACKGROUND OF THE INVENTION In recent years, with respect to health foods, so-called dietary fiber, polyphenols, and the like have attracted high interest.
These are chemical substances originally contained in relatively easily available materials, and many of them have been ingested without knowing them since ancient times. For example, polyphenols in chocolate, which has been promoted in recent years, were originally contained in cacao and were unknowingly eaten by people even before they were overtly disturbed. The same is true for polyphenols in wine, as is the fiber in konjac and burdock. Similarly, chemicals that were relatively accessible, readily available and inexpensive, but were left unnoticed for their chemical or biological activity, or were intended for entirely different uses There are countless. Considering that some of them may have unexpected antiviral and antibacterial properties, if such chemicals can be identified effectively, cheap and powerful medicinal substances can be stably and continuously provided to society. It is one of the ideas that started the present invention that we thought could bring the gospel to mankind.

On the other hand, in recent years, viruses such as AIDS and influenza have spread and afflicted mankind around the world. In response to this, researchers have made tremendous research efforts. However, satisfactory medicinal substances are not readily developed. Absent. The same applies to bacteria. In particular, synthetic new drugs, even if they have a strong effect, also have strong side effects, often producing undesirable results of suppressing pathogenic microorganisms and exhausting the human body. In addition to synthetic substances, so-called natural products are also known to have anticancer, antiviral, and antibacterial properties, and these are also used for folk remedies. Despite the benefits of natural products, there is concern about the stable and continuous supply of them in quantity and quality, and there is a problem in widespread use in society.

[0004]

SUMMARY OF THE INVENTION Accordingly, the present invention examines a number of chemicals that can be supplied relatively stably and continuously, have few side effects, and exhibit desirable antimicrobial activity. It has been made with a task to be determined and used as an antimicrobial agent.

[0005]

Means for Solving the Problems In order to solve this problem, the present invention first selects compounds having a benzene ring broadly, focuses on phenols among them, and further has a hydroxyl group and a methoxy group. The antiviral activity and the antibacterial activity were evaluated and confirmed by aiming at chemical substances, particularly guaiacol and lignins, and chemical substances related thereto. The virus targets two AIDS viruses and three influenza viruses, and the fungi target Escherichia coli O-157, Klebsiella pneumoniae, S. enteritidis (Salmonella), Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis. did. Although the chemical substances taken up in the initial test of the present invention were in excess of 100 kinds, lignins were tested on a very large number of them. From that process, even the same substance that is enclosed by the word "lignin" has characteristics and is not uniform, and even if it is effective, for example, it works for AIDS but not for influenza, or for viruses Works on bacteria but not on bacteria, or conversely, works on bacteria but does not work on AIDS. Also, the same AIDS virus has an inhibitory effect on the third day of culture, but the virus concentration rises on the 6th. There were various situations such as no inhibitory effect on the eyes (no activity on the 6th) (specifically, the main ones will be described in detail later). After all, each substance is examined for its properties, its molecular weight is measured, and its constituent elements are analyzed. The present invention has been made by strictly and strictly selecting that some chemical substances (eg, lignins) are effective or ineffective. Those which are determined to be useful through such enormous work / test / research / consideration and which are disclosed in the present invention are the 14 chemical substances (shown in the chemical formulas 1 to 14) described in detail below (hereinafter referred to as chemical substances). , These are referred to as samples 1 to 14) and their related substances.

Description of Samples The samples disclosed in the present invention are the following 14 types. (1) Guaiacol This substance is also called 1-hydroxy-2-methoxybenzene, 2-methoxyphenol, o-methoxyphenol, o-hydroxyanisole, 2-hydroxyanisole, methylcatechol, etc., in addition to guaiacol. This structure is shown below.
It is represented by the following equation.

Embedded image This guaiacol has been conventionally produced in large quantities as an industrial material, and was usually used for applications such as analytical reagents and preservatives. Guaiacol used in the present invention was manufactured by West West Milwaukee, Wisconsin, USA.
98% purity manufactured and sold by Aldrich Chemical Company, Inc. of St. Paul 1001
Guayacol. Its properties are colorless or slightly yellowish liquid, boiling point 205 ° C, melting point 17-29 ° C, flash point 82 ° C, specific gravity 1.129. As described above, in view of the conventional use of this substance, it was very unlikely that the substance had antiviral properties, in particular, anti-AIDS activity or anti-influenza activity, but these activities were found by the present invention. It is surprising that it was confirmed and found to have multifaceted antimicrobial properties including antibacterial properties. In selecting guaiacol, the present inventor paid attention to potassium guaiacol-4-sulfonate 0.5 wettable powder as an analog. This is made by adding potassium to guaiacol to make it easily soluble in water. However, when an antiviral test was carried out using this analog, cell damage occurred and measurement became impossible. Even though they are similar, simple inferences must be strictly discouraged, as small differences in components can make a big difference in results. In addition to the synthetic products mentioned above, guaiacol is a natural guaiac tree (Guaiacum).
sanctum). The stem is cut to about 1 meter and a vertical hole is made to elute the components by direct heat, or a small piece of wood (chip) is extracted by boiling. Sap may be leached by damaging the standing tree. In any case, "guaiac fat" collected from guaiac trees is known to be used today as an antioxidant for foods, as a reagent for detecting oxidants and oxidases, and as a reagent for analysis. Therefore, it is a product that is industrially produced. This guaiac fat was tested for anti-AIDS activity together with guaiacol in the present invention and produced equal or better results. This will be described in detail in a test example described below (see the section of guaiacol (sample 1) in "100% growth inhibitory activity of AIDS virus (1)").

(2) Lignin sulfonic acid The structure of this substance is represented by the following formula.

Embedded image (In this formula, R ₁ is mainly OCH ₃ , R ₂ is H or another lignin unit, and R ₃ is another lignin unit.) This lignin sulfonic acid is obtained from Kanto Chemical Co., Ltd. This product is manufactured and sold in Nihonbashi-Honmachi 3-2-8) and has the property of black powder with a thin vanilla odor. The results of measuring the molecular weight distribution of this substance by the measurement method described later are shown in the graph of FIG. 2, and most of the substances have a molecular weight of 20,000 to 45,000. It was confirmed that this substance also has excellent antiviral properties (anti-AIDS properties, anti-influenza properties) and antibacterial properties according to the present invention. The present inventor focused on lignin alkali (manufactured by Aldrich) at first among so-called lignins and carried out a method (culturing for 3 days,
(Cultured for 6 days). Lignin alkali is a form in which H (or another lignin unit) is added in place of SO ₃ H in the formula of chemical formula 2 (lignin sulfonic acid), and R ₁ and R ₂ are exchanged. This is very closely related to ligninsulphonic acid and would normally be expected to be equally effective, but in fact it was not. The test results showed that, contrary to expectation, there was no "day 6 activity" (for the significance and importance of day 6 activity, see "100% growth inhibitory activity of AIDS virus" described later (Table 2). Will be described). From this, it is understood that the validity cannot be correctly determined by mere guesswork or prediction with the color glasses of "common sense", and that the truth must be actually tested, considered, and confirmed individually.

(3) 2,6-dimethoxyphenol The structure of this substance is represented by the following chemical formula 3.

Embedded image This substance is a liquid product sold by Wako Pure Chemical Industries, Ltd. (Doshu-cho, Chuo-ku, Osaka-shi). It is brownish liquid and has a strong smell like oxyfur. The anti-AIDS virus and antibacterial properties of this substance are comparable to guaiacol.

(4) 3,5-dimethoxyphenol The structure of this substance is represented by the following chemical formula 4.

Embedded image This substance is a product manufactured by Aldrich of the United States mentioned above,
It is an off-white crystalline powder with a purity of 99%. Boiling point 17
It has a melting point of 2 to 175 ° C, a melting point of 45 to 47 ° C, and a flash point of 78 ° C. Irritating (eye, skin, respiratory system, etc.). This substance has been found to have a broad and versatile efficacy with anti-AIDS, anti-influenza, and antimicrobial properties.

(5) Lignosulfonic acid sodium salt (Lig
nosulfonic acid, sodium salt) The unit structure of this substance (also called sodium ligninsulfonate) is represented by the following chemical formula (5).

Embedded image (In this formula, R ₁ is mainly H or another lignin unit, R ₂ is mainly OCH ₃ , and R ₃ is mainly another lignin unit.) This substance is manufactured and sold by Aldrich of the United States. Substance. This lignosulfonic acid sodium salt is
It is a sulfonated lignin polymer mainly isolated from pulp mills using European spruce (Norway spruce) as a raw material. Ion exchange (from calcium to sodium) and filtration are used for isolation. The branched macromolecular structure of lignosulfonic acid sodium salt contains sulfonate groups (sulfonation degree is 0.46 per phenylpropane repeating unit, corresponding to 6.7% sulfur content and 5.5% sodium content). ). Lignosulfonic acid sodium salt includes primary and secondary aliphatic OH and phenolic OH. The adjacent phenylpropane repeat units are linked via a CO-C as well as a C = C bond. The methoxy content is 10.8% calculated, 46.17% carbon and 4.70% hydrogen by elemental analysis. It is a free-flowing (smooth) non-toxic powder with a bulk density of 0.
5 g / cm ₃ . Soluble in water. The color is brown. It smells like thin 'jintan'. Conventionally, this substance has been used exclusively as an industrial material as an anionic surfactant and as an additive to phenol formaldehyde resin, so that the antiviral and antibacterial properties as noted by the present invention were not known at all. . However, it is surprising that the present invention has been found to have very good anti-AIDS virus properties, anti-influenza virus properties, and certain antibacterial properties. The molecular weight distribution of this substance measured by the measurement method described below is as shown in the graph of FIG.
Most of the molecular weight is 20,000, molecular weight 12,500 and 3,800
Is also found to be present.

(6) Lignosulfonic acid, sodium salt, acetate The unit structure of this substance is represented by the following chemical formula (6).

Embedded image (In this formula, R ₁ is mainly H or another unit, R ₂ is mainly OCH ₃ , and R ₃ is mainly another unit.) This substance is also produced and sold by Aldrich of the United States as above. It is a derivative polymer of lignosulfonic acid sodium salt. It is prepared from a homogeneous phase solution using acetic anhydride as a reagent. Contains aliphatic and aromatic acetoxy groups. Each phenylpropane repeating unit has 0.46 sulfonate groups. The adjacent phenylpropane repeat units are linked via C = C and C-O-C bonds. Elemental analysis shows 47.16% carbon, 4.72% hydrogen, 3.77% nitrogen
% In the ICP assay is S5.4% and Na3.1%. Aliphatic OH predominates over phenolic OH. The molecular weight measured in the same manner as above is shown in the graph of FIG.
It is recognized as 18,000. It is a smooth, non-toxic yellow-brown powder with a slightly weak acetate (acetate) odor. Soluble in water at any pH, but insoluble in most organic solvents. Usually, it is used as an industrial material as a hydroxyl-free water-soluble material which is modified into a derivative soluble in an organic solvent by a chemical treatment. Of course, use as a medicinal ingredient such as antiviral properties has not been expected at all. However, according to the present invention,
(5) Like lignosulfonic acid sodium salt, it was found to have versatile effects such as excellent anti-AIDS virus and anti-influenza virus properties and a certain antibacterial property.

(7) Lignin, organosolve (Lignin,
organosolv) The unit structure of this substance is represented by the following chemical formula 7.

Embedded image (In this formula, R ₁ is OCH ₃ or H or another unit, R ₂ is mainly OCH ₃ , R ₃ is OH or another unit, and R ₄ is another unit.) Similarly, it is a chemical manufactured and sold by Aldrich, USA. It is a polymer lignin isolated from ordinary pulp mills using a mixture of hard wood (50% maple, 35% hippo, 15% poplar) as a raw material. It contains primary and secondary aliphatic OH and phenolic OH.
6.5%, hydrogen 6.1%, OCH ₃ 18.9%, sucrose 0.5% or less, ash content 1% or less. It is a non-toxic powder that flows smoothly and has a light alcoholic odor. It is soluble in alkaline aqueous solutions and certain organic solvents. Conventionally, it has been usually used industrially as an additive for phenol formaldehyde resin, but other uses have not been known. According to the present invention, some anti-AIDS virus properties were recognized.

(8) Lignin, hydrolytic (Lig)
nin, hydrolytic) The unit structure of this substance is represented by the following chemical formula 8.

Embedded image (In this formula, R ₁ is OCH ₃ or H or another unit, R ₂ is H or OCH ₃ , R ₃ is mainly another unit, and R ₄ is mainly another unit.) Is a chemical substance manufactured and sold by Aldrich, Inc. of the United States, and is an auto-hydrolyzable polymer lignin mainly isolated from the hydrolysis process using Sugar Cane Bacchus as a raw material. Branched macromolecular structures include primary and secondary aliphatic OH and more prevalent phenolic OH. The methoxy group content ranges from 9 to 11%. Linking to a neighboring phenylpropane repeating unit via a C ＝ C, C—O—C bond. It is a smooth, non-toxic brown powder that is soluble in aqueous alkaline solutions and certain organic solvents such as methanol or ethanol + acetone, methylene chloride, chloroform or benzene. It is usually used as an additive for phenol formaldehyde resin. According to the present invention, the anti-AIDS virus property of this substance was partially recognized.

(9) Lignin, Organosolv, Acetate The unit structure of this substance is represented by the following formula.

Embedded image (In this formula, R ₁ is H, OCH ₃ or another unit, R ₂ is mainly OCH ₃ , R ₃ is mainly another unit.) This is also a chemical substance manufactured and sold by Aldrich Corporation of the United States. It is a polymer derived from organosolve.
The branched macromolecular structure contains primary, secondary aliphatic and aromatic acetoxy groups. Small amount of CO, COOH
Including groups. It is a non-toxic, light brown powder that flows smoothly and is soluble in most organic solvents but insoluble in water. This substance is used as a thermoplastic material added to other polymer materials as a viscosity modifier, a colorant, or the like. According to the present invention, some anti-AIDS virus properties were recognized.

(10) Lignin, hydrolytic,
Hydroxymethyl derivatives (Lignin, hydrolitic, hrdro
xymethyl derivative) The unit structure of this substance is
0.

Embedded image (In this formula, R ₁ is mainly OCH ₃ or another unit, R ₂ is mainly OH or another unit, and R ₃ is mainly another unit.) This is also a product of Aldrich, and ) Is a derivative polymer of lignin and hydrolytic. It is prepared by treating lignin with formaldehyde in a homogeneous solution (aqueous alkaline solution). It contains primary, secondary aliphatic and phenolic OH groups and a small amount of carboxy functions. The property is a non-toxic black powder that flows smoothly,
It is soluble in alkaline aqueous solutions and certain organic solvents. This material has previously been used as an additive to phenol formaldehyde resins. According to the present invention, anti-AIDS virus activity was partially recognized.

(11) Lignin, organosolve, propionate The unit structure of this substance is represented by the following formula.

Embedded image (In this formula, R ₁ is mainly H or OCH ₃ or another unit, R ₂ is mainly OCH ₃ , R ₃ is mainly another unit.) This is also a chemical substance manufactured and sold by Aldrich, USA, and has a molecular weight distribution. Is as shown in the graph of FIG. Molecular weight 15,000
Is the center, and molecular weights of 9,200, 7,500, and 1,650 are mixed. This is a derivative polymer of the lignin organosolve (7), and is produced by a homogeneous phase reaction of lignin in propionic acid using propionic anhydride as a reagent and sodium propionate as a catalyst. Prior to isolation, it is partially bleached with H ₂ O ₂ to a brown powder. 57.39% carbon by elemental analysis,
5.58% hydrogen. It is a non-toxic powder that flows smoothly. Conventionally, colorants for polymerizable materials and plastics,
It is used as a soluble thermoplastic lignin derivative for purposes such as viscosity modifiers. According to the present invention, lignin organosolv propionate was found to be useful in anti-AIDS virus and antibacterial properties.

(12) 4-Benzyloxyguaiacylglycerol-β-guaiacylether The structure of this substance is represented by the following formula.

Embedded image This substance is a chemical supplied by Hokkaido University. The properties are whitish powder. According to the invention, this substance was found to have some anti-AIDS and anti-influenza virus properties. The SOS related to this substance will be described later.

(13) Syringaldehyde The structure of this substance is represented by the following formula.

Embedded image This substance is also a chemical manufactured and sold by Aldrich, USA. According to the present invention, syringaldehyde has a considerably strong anti-AIDS property, but also has strong cell damage. Therefore, it is desirable to reduce the damage by using a mixture with other chemical substances when applying.

(14) Veratryl alcohol The structure of this substance is represented by the following formula.

Embedded image This substance is often used as a simplified model of lignin units. In this sense, when tested as an analog of lignin according to the present invention, antiviral properties were found.

Among the various lignins described above, the following 4
The species and molecular weight determinations performed on humic acid (see "General Analysis of Lignin" below (Table 1)) in connection with the species are described below. [1] Reagents / equipment (1) Test lignin 1. Lignin sulfonic acid 2. Lignin sulfonic acid sodium salt 3. Lignin sulfonic acid sodium salt acetate 4. Lignin organosolv propionate (5. humic acid) (2) Reagent 1. Blue Dextran 200 (Pharmacia Fine Chemicals) 2. Polyethylene glycol 400, 1000, 4000, 6000, 20
000 (Wako first grade) (abbreviated as PEG) 3. Methanol (Wako special grade) (The molecular weight distribution of reagent 1.2 is shown in the graph of Fig. 1.) (3) Equipment used 1. Fraction collector (ADVANTEC SF-2120) Operating conditions Simple Mode Wait time 0 minutes Fractionation 85 drop / tube (molecular weight marker) 120 drop / tube (lignin) (both 2.5 ml tubes) 2. Spectrophotometer (BECRUN DU 7400) 3. Brix meter (ATAGO HAND REFRACTOMETER N-1E) (4 ) Column Gellot agent Toyopearl HW50F (Tosoh) Column with cock (φ2.5 × 120 cm, glass) [2] Method (1) Column preparation The column was fixed vertically to a stand. After washing the gel filtration agent with deionized water, the column was packed into the column according to an ordinary method by gravity. The upper part of the column was connected to the solvent tank installed above, and the column was thoroughly washed with three times the amount of deionized water of the gel filtration agent. The cock at the lower end of the column was connected to the fraction collector. (2) Elution of Molecular Weight Markers Five kinds of molecular weight markers (blue dextolan) and PEG were dissolved in deionized water so as to be about 5 to 10%, respectively. The deionized water in the column was dropped to the tip of the gelling agent, and the lower cock was closed once. Then, 5 ml of the marker solution was added to the column, taking care not to disturb the surface of the gelling agent. The lower cock was opened to drop the additive solution to the gel filtration agent surface, and then elution was started with deionized water.
The eluate was fractionated by a fraction collector. Each fraction was analyzed by a spectrophotometer (blue dextran, measurement wavelength 59
5 nm), elution of the marker was detected with a Brix meter (PEG), and an elution curve was prepared. A molecular weight curve was created using the maximum elution fraction as the elution position of each marker. (3) Elution of lignin The column was washed and equilibrated with 50% methanol / deionized water (v / v) twice as much as the gel peroxidant. About 20 mg of each lignin was dissolved in a 50% aqueous methanol solution and added to the column.
Elution was performed with 50% methanol in the same manner as for the molecular weight marker, and the eluate was fractionated. The elution of lignin in each fraction was measured using a spectrophotometer. The measurement was performed at the maximum absorption wavelength of each lignin solution. Lignin measurement wavelength (nm) 1. Lignin sulfonic acid 275 350 2. Lignin sulfonic acid sodium salt 275 305 3. Lignin sulfonic acid sodium salt acetate 275 310 4. Lignin organosolv propionate 275 290 Prepare elution curve of each lignin To determine the maximum elution fraction, and prepare the molecular weight curve (each lignin 1. ~ 4.
Was estimated from the respective graphs of FIGS. 2 to 5). [3] Result 1. Lignin sulfonic acid: molecular weight of 20,000 or more and 45,000 or less, mostly 11,000
There are molecules. 2. Lignin sulfonic acid sodium salt… Molecular weight of 20,000 is mostly and molecules of molecular weight of 12500 and 3800 are also present. 3. Lignin sulfonic acid sodium salt acetate… Molecular weight 18000 4. Lignin organosolve propionate… Molecular weight 15,000 is the center, and molecular weights 9200, 7500 and 1650 are mixed. 5. Molecular weight of humic acid: For humic acid, the molecular weight of humic acid was estimated in the same manner as in the analysis of the molecular weight of lignin. However, 30 mg of humic acid was dissolved in 5 ml of a 0.1 N aqueous sodium hydroxide solution and eluted with a 0.1 N aqueous sodium hydroxide solution. Fraction collector condition: Simple mode 75drop /
Fr. (25 ml) As shown in the graph of FIG. 6, there was an elution peak at Fr. No. 117, and the molecular weight of humic acid was determined to be about 3500 from the molecular weight calibration curve.
It was estimated.

The general items of lignin and other natural lignin used in the present invention and humic acid in relation to the lignin have been analyzed, and their contents are described below. The samples used are as follows. Specimen 1) ligninsulfonate (sample 2) 2) lignosulfonic acid sodium salt (sample 5) 3) lignosulfonic acid sodium salt acetate (sample 6) 4) lignin organosolv propionate (sample 11) of A mushroom sulfate Lignin fraction B Lignin hydrochloride fraction of mushroom C Humic acid [1] Method Each sample (C will be described later) was accurately measured to about 200 mg / ml and dissolved in deionized water. After thorough stirring, the mixture was centrifuged at 3000 rpm for 15 minutes to separate the supernatant, which was subjected to the following test. [2] Test (1) OD and pH at 500 nm A 1% (W / V) aqueous solution of each sample was prepared, and the absorbance and pH at 500 nm were measured by a conventional method. (2) Protein The amount of protein in each sample aqueous solution was measured by the method of Bradford. 20 μl of the sample solution was placed in a 1.5 ml test tube, mixed with 1 ml of the Bradford solution, allowed to stand at room temperature for 5 minutes, and the absorbance at 595 nm was measured. A control obtained by adding 20 μl of deionized water to a Bradford solution was used as a control. The sample solution was measured after dilution with an enemy machine, and care was taken not to be affected by sample coexisting substances. The amount of protein in the sample solution was calculated from a calibration curve prepared using bovine serum albumin as a standard sample.
It was converted to the amount of protein per g. (3) Glucose The amount of glucose in the sample was measured using Glucose C-II Test Wako (Wako Pure Chemical Industries). This measurement reagent is an enzyme method reagent and has high specificity. A 20 μl sample solution was placed in a test tube, and 3.0 ml of a coloring reagent was added and mixed. After heating at 37 ° C. for 5 minutes, the absorbance at 505 nm was measured. As a control, a solution obtained by adding 3.0 ml of deionized water to each sample solution was used to remove the influence of the color of the sample solution. At the same time, a standard curve was prepared by reacting a glucose standard solution. The amount of glucose in the sample solution was calculated from the calibration curve and converted to the amount of glucose per 1 g of the sample. (4) Total sugar The total amount of sugar in the sample was measured by the phenol-sulfuric acid method.
Dispense 200 μl of sample solution into a test tube, and add 5% phenol solution
Add 200 μl. Then, 1 ml of concentrated sulfuric acid was added dropwise, followed by rapid stirring. After standing at room temperature for 20 minutes, the absorbance at 490 nm was measured. As a control, distilled water was reacted in the same manner and used.
At the same time, a standard curve was prepared by reacting glucose as a standard solution. The total sugar in the sample solution was calculated from the calibration curve as the amount of glucose, and converted to the amount per 1 g of the sample. The results of each of the above analysis tests are shown in Table 1 below.

[0022]

TABLE 1 General item analysis 1% aqueous solution of the protein glucose total sugar specimen 500NmOD pH of such lignin (mg / g) (mg / g) (mg / g) 1) ( specimen 2) 1.405 (× 10) 8.87 61.36 0.0 357.1 2) (Sample 5) 0.579 8.79 19.19 4.56 239.1 3) (Sample 6) 0.347 4.00 24.68 0.27 224.8 4) (Sample 11) 0.300 4.86 4.38 0.44 0.399 A (sulfuric acid fraction) 0.024 3.24 (0.36) 0.35 0.323 B (hydrochloric acid) (Fraction) 0.014 3.12 (0.43) 0.28 0.318 C (humic acid) 0.030 5.36 1.30 0.18 0.141

In the present invention, as described above, more than 100 kinds of candidate substances were selected and intensive tests and researches were conducted on antimicrobial properties. From the above, it was carefully selected and found to be effective or partially effective. There are 14 chemicals. The tests performed were 100% inhibitory activity against AIDS virus growth, 100% inhibitory activity against influenza virus growth, and antimicrobial activity as described below.
The 0% growth inhibitory activity is described below.

AIDS virus 100% growth inhibitory activity : The test method is as follows. Each sample (more than 100 kinds of the above-mentioned chemical substances) is dissolved in water to prepare an aqueous sample solution. The initial concentration of the aqueous solution is 1 mg / ml (1 mg of sample powder per 1 ml of aqueous solution =
1000 μg). AIDS virus (HIV-1 and HIV-2
2) and a suspension of MT-4 cells. Using a microplate having twelve wells (holes), the sample aqueous solution is diluted at a concentration of 1000 μg / ml in the first well, and then diluted two-fold from the second well. That is, the sample concentration of the 2 wells was diluted to 500 μg / ml, 3 wells to 250 μg / ml, 4 wells to 125 μg / ml, 5 wells to 62.5 μg / ml, and finally 0.49 μg / ml (2048) for the 12 wells. (Double dilution). For easy understanding, the table shows the sample concentration (A) (μg / ml) and dilution factor (B) of each well. Well 1 2 3 4 5 6 7 8 9 10 10 11 12 A 1000 500 250 125 62.5 31.3 15.6 7.81 3.91 1.95 0.97 0.49 B1 24 8 16 32 64 128 256 512 1024 2048

In each well of the microplate, 100 μl of an aqueous sample solution of each concentration described above and an AIDS virus suspension 10
Inject 0 μl each. The AIDS virus coexisting with the specimen was cultured in each well, and all the wells were observed on the third and sixth days of culture, and the growth of AIDS virus was reduced by 10%.
Determine the 0% blocking well. 100% growth inhibition means that MT-4 cells coexisting in the wells are healthy without being destroyed or deformed by the AIDS virus. On the third day of culture, the virus concentration is 10 TCD
However, when the cells are cultured until day 6, the virus concentration of the control increases to 100 TCD. "Day 6 activity", which suppresses virus growth by 100% with this increasing concentration,
Therefore, it is an important criterion for evaluating the validity / invalidity of the sample. From this evaluation test, those showing remarkable results were selected, and among them, samples 1 to 7 (the above-described chemicals 1 to 7) were selected.
Are shown in Table 2 below. In Table 2, it is shown that AIDS growth was inhibited by 100% up to the wells marked with a circle (the numbers in parentheses indicate the sample concentration at that time in μg /
ml), and to the right (higher numbered) wells, 100% did not block virus growth. Further, in the wells marked with *, it is indicated that cell damage was caused not by the virus but by the test compound itself.

[0026]

[Table 2] 100% growth inhibitory activity of AIDS virus (1) (Samples 1 to 7 correspond to Chemicals 1 to 7) Well number Sample culture 12 3 4 5 6 7 8 9 10 11 12 (abbreviation) 13 Sun * ○ (31.3) 6T ₃ 6th * ○ (125) 4T ₃ 23 days * ○ (7.81) 8T ₄ 6th * ○ (15.6) 7T ₄ 33 days * ○ (31.3) 6T ₄ 6th * ○ (31.3) 6T ₅ 4 3 days * ○ (125) 4T ₂ 6th * ○ (125) 4T ₃ 5 3 days ○ 10T ₀ 6th ○ 9T ₀ 6 3 days ○ 9T ₀ 6th ○ 10T ₀ 73 days * ○ 7T ₅ 6 days * <<6T ₅ Sample 1 = Guaiacol Sample 2 = Lignin sulfonic acid Sample 3 = 2,6-dimethoxyphenol Sample 4 = 3,5-dimethoxyphenol Sample 5 = Lignosulfonic acid sodium salt Sample 6 = Lignosulfonic acid sodium salt Sample 7 = Lignin organo Solve

Thus, guaiacol (specimen 1) increased the AIDS virus growth in 6 wells (31.3 μg / ml) after 3 days of culture.
, A 100% inhibition up to 4 wells (125 μg / ml) in 6-day culture (activity on day 6). Cell damage was observed up to 3 wells (250 μg / ml). As shown on the right side of Table 2, the results were 6T ₃ (activity on day 3), 4T ₃
Abbreviated as T ₃ (activity on day 6) (T stands for Toxicity). In the present invention, in addition to guaiacol (sample 1), guaiac oil containing guaiacol (extract from a natural guaiac tree) was tested for its anti-AIDS effect. Result is,
Day 3 activity 3.9 μl / ml (cell damage 7.8 μl / ml) (abbreviated 9T ₈ ), day 6 activity 3.9 μl / ml (cell damage 7.8 μl / m
l) A good result of (9T ₈ ) was obtained. Therefore, in the present invention, guaiac fat can be said to be equivalent to guaiacol at least in terms of antiviral properties, particularly anti-AIDS virus properties, and although it is a natural product, it can be easily supplied as an industrial product as described above. This is also an object of the present invention, as is Guaiacol.

The lignin sulfonic acid of the specimen 2 was cultured in 8 wells (7.81 μg / ml) for 3 days and 7 wells (15.
(6 μg / ml), respectively, with 100% inhibition and cytotoxicity up to 4 wells each. Day 3 activity 8T ₄ ,
Figure of 6 day activity 7T ₄ shows that an extremely excellent anti-AIDS virus agents. 2,6- of sample 3
100% inhibitory activity of dimethoxy phenol, day 3 activity 6T _4, 6 day activity is a 6T _5, strong cytotoxicity. The same is true for the anti-influenza virus described below.The ability to suppress the virus itself is not weak, but it does not exert a higher effect because it damages the cells culturing the virus itself. That is. However, the fact that the 6-day activity of 6 wells is achieved is an excellent effect that has not been foreseen so far, and thus can be used as a useful anti-AIDS agent.

The 3,5-dimethoxyphenol of sample 4 is inferior to sample 3, but still has 100% inhibition of the AIDS virus at a concentration of 4 wells (125 μg / ml) at day 6 activity. The activity of 4T ₃ on day 6 causes cell damage in 3 wells immediately before the 100% inhibitory effect. In such a case, an appropriate amount (for example, 1: 1) of 3,4-dimethoxyphenol ( When a methoxy group is attached to the 3-position and 4-position in the formula of Chemical Formula 4, the concentration of each component is reduced by half,
It was observed that the cytotoxicity was reduced to 1 well without decreasing the 0% inhibitory effect. Other substances such as albumin may be mixed instead of 3,4-dimethoxyphenol. Such a reduction effect is generally effective for other samples, and by mixing two or three agents,
It was recognized that cytotoxicity can be reduced without diminishing the respective virus-suppressing effects. Although not directly an object of the present invention, referring to the lignin fractions derived from mushrooms described above in "General item analysis of lignin" (Table 1), these lignin fractions were also excellent. have been confirmed by the same test procedures that a beginning with various antibacterial activity anti-AIDS activity, albumin to reduce cytotoxicity caused case 1: the T ₃ by mix at a ratio of 1 T An effect of reducing to ₁ has been found (the identity of these lignin fractions is currently being identified by the present inventors).

The 100% inhibitory effect of sample 5, ie, lignosulfonate sodium salt, and sample 6, ie, lignosulfonate sodium salt acetate, in Table 1 is surprising.
10T ₀ means that at a concentration as low as 10 wells (1.95 μg / ml), virus growth is inhibited by 100% and cell damage is not caused. Even in the 9T ₀ 3.91μg /
100% inhibition at low concentrations of ml, with zero damage.
This suggests that both of these substances are extremely good anti-AIDS agents, and that they can be used in foods and drinks to produce non-toxic anti-AIDS foods and drinks. This is expected. Both of these substances show extremely high efficacy in anti-influenza virus properties described below, and antibacterial properties are also expected to a considerable extent. Therefore, one agent (one substance) has antiviral properties (AIDS and influenza) and antibacterial properties. A multifaceted antimicrobial agent (food,
(Beverage), it is strongly expected to be welcomed with high multi-sided utility that is frequently used on a daily basis.

In the 6-day culture of the specimen 7 (Chemical formula 7 = lignin organosolv) in Table 2, 100% inhibition was not achieved in the well 6;
Indicates 6 wells or less (<). However, in reality, cell damage is caused in well 5 (<6T ₅ ), so that it is practically difficult to use this substance as it is to inhibit the 100% growth of AIDS virus. However, as described above, cell damage can be reduced by using it in combination with other agents.

Samples other than the above, that is, chemical formulas 8 to 14
For the chemical substances corresponding to the above, the test results of 100% inhibitory activity using a microplate as described above are shown in Table 3 below using the above-mentioned abbreviation method.

TABLE 3 test body 3 days culture 6 days culture 8 lignin, hydro Li tick <6T ₅ 5T ₄ 9 Lignin, Organosolve, Acetate 6T ₅ <6T ₅ 10 Lignin, hydro Li ticks, hydro <6T ₅ 3T ₂ Kishimechiru derivatives 11 lignin, organosolv, propionate 6T ₅ 5T ₄ 12 4-Benzyloxy grayed cradling Le glycerol 5T ₃ 4T ₃ -Β-guaiacyl ether 13 syringaldehyde 11T ₈ 10T ₉ 14 Veratryl alcohol 5T ₄ 4T ₃

The sample 11 (lignin organosolv propionate) has a well of 100% inhibition and a well of cell injury, but the combination of 5% or 5% with the above-mentioned other agent having an effect of reducing the injury is used. Well (62.5μg / ml)
Utilizing the 100% inhibitory activity (activity on day 6), it is possible to obtain a sufficiently useful anti-AIDS virus substance. This lignin propionate is also effective in antibacterial properties, as described later, and thus can be a multifaceted antimicrobial agent, similarly to the above-mentioned specimens 5 and 6.

In connection with sample 12 (4-benzyloxyguaiacylglycerol-β-guaiacylether),
So-called SOS (syringyl glycerol-β-syringyl ether, also known as 3,5-dimethoxy-4-hydroxyphenylglycerin-β- (2,6-dimethoxyphenol) ether) having a similar lignin model structure When the anti-AIDS virus activity was tested in the same manner, the activity on the third day was 15.6 μl / ml (cell damage 31.3 μl / ml)
(7T ₆ ), but unfortunately no activity was obtained on day 6. On the other hand, 4-benzyloxyguaiacylglycerol-β-guaiacylether (sample 12), which is also a model for lignin, has an activity of 5 days.
T ₃ and 125 μl / ml (4T ₃ ) of activity on the 6th day were obtained, and it was confirmed that they had excellent anti-AIDS activity. From this, it is understood that it is incorrect to infer lightly that all lignins have the same effect.

The syringaldehyde in sample 13 has a remarkable anti-AIDS virus property in that it achieves 100% growth inhibition at a low concentration of 10 wells (concentration: 3.91 μg / ml) in activity on day 6. Is recognized. Syringaldehyde is also effective in the anti-influenza virus properties described below, and also exhibits antibacterial properties depending on the concentration, and thus can be said to be a multifaceted antimicrobial agent. As for the other specimens 8, 9, and 10, the usefulness as an anti-AIDS agent is hard to be recognized as it is, similarly to the specimen 7 (lignin organosolve). As a reference, a similar test at the cell level was also performed for potassium guaiacol-4-sulfonate 0.5 wettable powder, but this was not found to be useful as an anti-AIDS virus agent.

In general, the mechanism by which the above-mentioned various chemical substances according to the present invention exhibit anti-AIDS virus properties is not intended to limit the present invention thereto. In other words, it is presumed that the chemical substance of the present invention physically inhibits the action of the protease produced by the AIDS virus from attaching to the target cells, by physically removing the protease from the target cells. Similarly,
It is presumed that the chemical substance of the present invention has a similar inhibitory effect mechanism for influenza virus described below.

Next, among the chemical substances of the present invention, those which have been found to be useful in the anti-AIDS virus activity test as described above, ie, guaiacol (Chem. 1), ligninsulfonic acid (Chem. 2), 2,6 -Dimethoxyphenol (formula 3),
3,5-dimethoxyphenol (chemical formula 4), ligninsulfonic acid sodium salt (chemical formula 5), ligninsulfonic acid sodium salt acetate (chemical formula 6), lignin organosolv propionate (chemical formula 11), 4-benzyloxyguaiacyl Glycerol-β-guaiacyl ether
An evaluation test was performed on 2), syringaldehyde (Chemical Formula 13), and veratryl alcohol (Chemical Formula 14) to evaluate the effect of suppressing influenza virus growth. The viruses used were: 1) Influenza virus A / Hokkaido / 1/96 (H1N1) (hereinafter abbreviated as H1N1) 2) Influenza virus A / Hokkaido / 1/97 (H3N2) (hereinafter abbreviated as H3N2) 3) Influenza Virus B / Hokkaido / 1/97 (hereinafter abbreviated as type B).

Preparation of MDCK Cell Maintenance Solution An MDCK cell maintenance solution is prepared to maintain MDCK cells for inoculating influenza virus. Composition of the maintenance solution : 1. Basal medium: 88 ml of Eagle's MEM 2. Penicillin and stomay: 200 units / ml each (1 ml of 20,000 units / ml solution is added to 100 ml) 3. Glutamine: 0.03% added (3% solution 4. Add glucose: 0.01% (add 1 ml of 1% solution to 100 ml) 5. Add bovine albumin (fraction V): 0.2% (10%
(Add 2 ml of solution to 100 ml) 6. Add vitamin: 4% (add 4 ml of 100-fold solution to 100 ml) 7. Modify pH: Modify pH to 7.6-7.8 (10% of 5% solution)
(Put almost 3ml in 0ml)

Each of the test substances (Chemical Formulas 1 to 6, 11 to 14) is serially diluted with the above-mentioned culture solution of MDCK cells. The initial concentration (stock solution) of each sample is 10 mg / ml (liquid,
Guaiacol was 10 μl / ml and 2,6-dimethoxyphenol was 2 μl / ml-* in Table 4. Prepare a large number of test tubes, prepare the sample stock solution, and serially dilute up to 128-fold (in addition to 2-fold serial dilution, 2.6-fold dilution, 5.3-fold dilution, 10-fold dilution, and 100-fold dilution) 0.2 The ml is mixed with 0.2 ml of 10 TCID virus solution into 1 ml of culture in a number of tubes containing cells. In the determination method, when the virus grew to 100 TCID (3 to 4 days after inoculation), the dilution concentration at which the sample suppressed virus growth was defined as an effective amount. The minimum concentration shows a (MIC) (in parentheses the growth of the influenza virus in the following Table 4 was 100% blocked by the same abbreviations, for example, a 3.5T _1, tube No.3 (4-fold dilution ) And No.4 (8-fold dilution)
% Inhibition was observed).

[0040]

(Table 4) Minimum inhibitory effective dose (MIC) (Sample numbers correspond to Chemicals 1 to 6, 11 to 14) Influenza virus sample H1N1 H3N2 B 1 3.8 μl / ml (2.5 T ₁ ) 1.9 μl / ml (3.5 T _{1) 3.8μl / ml (2.5T 1} ) 2 1.9mg / ml (3.5T 1) 1.9mg / ml (3.5T 1) 2.5mg (3T 1) * 3 1.0μl / ml or less (<2T ₁₎ to the left the same left same _{4 5.0mg / ml (2T 1)} 1.9mg / ml (3.5T 1) 3.8mg / ml (2.5T 1) 5 0.16mg / ml (7T 0) 0.10mg / ml (8T 0) 0.63mg / ml (5T ₀ ) 6 0.16 mg / ml (7T ₀ ) 0.16 mg / ml (7T ₀ ) 1 mg / ml (4.5 T ₀ ) 11 2.5 mg / ml (3T ₁ ) 2.5 mg / ml (3T ₁ ) 3.8 mg / ml (2.5T ₁ ) 12 8.0 mg / ml (2T ₁ ) 2.5 mg / ml (3T ₁ ) 2.5 mg / ml (3T ₁ ) 13 3.8 mg / ml (2.5 T ₁ ) 1.9 mg / ml (3.5 T ₁ ) _{1) 3.8mg / ml (2.5T 1} ) 14 3.8mg / ml (2.5T 1) 1.9mg / ml (3.5T 1) 3.8mg / ml (2.5T 1)

In Table 4, all the other specimens were obtained, except that 2,6-dimethoxyphenol had no initial influenza virus-suppressing effect, partly due to the initial concentration of 2 μl / ml. Has achieved a useful suppression effect. Also of particular interest here are lignosulfonic acid sodium salt (sample 5) and the same acetate (sample 6). That is, the sodium salt (sample 5) is H3N
2 Inhibition of influenza is inhibited by 100% at very low concentration of 100-fold dilution, and at worst, 100% inhibition is achieved at a low concentration of 16-fold dilution (No. 5 test tube) against type B. 7T ₀ (H1N1) of sodium salt acetate (sample 6)
And H3N2) are also amazing results, at worst 4.5 T ₀ (10-fold dilution) for type B. In addition, guaiacol (sample 1), lignin sulfonic acid (sample 2), 3,5-dimethoxyphenol (sample 4), lignin organosolve propionate (sample 11), 4-benzyloxyguaiacylglycerol-β-g Ayasyl ether (Sample 1
2), syringaldehyde (specimen 13), and veratryl alcohol (specimen 14) have also been shown to be useful as effective anti-influenza agents (food and beverages).

Subsequently, another suppression evaluation test for the same influenza virus will be described. The target influenza viruses are the same as before (1) H1N1, (2) H3N2, (3) B
Type, and the specimens are lignosulfonic acid sodium salt (specimen 5) and the same acetate (specimen 6). The difference from the previous test is that the initial concentration of the sample stock solution was 2 mg / ml (2 times). Surprisingly, no cytotoxicity occurred even at this stock concentration (zero). Table 5 shows the results (MIC)
Shown in

TABLE 5 Influenza virus suppression Evaluation (MIC) (2) (high concentration) influenza virus sample H1N1 H3N2 B 5 83.3μg / ml 10.4μg / ml 666.7μg / ml (24 fold dilution) (192-fold dilution) (3 times (Dilution) 6 83.3 μg / ml 125 μg / ml 666.7 μg / ml (24-fold dilution) (16-fold dilution) (3-fold dilution) It is noted that even if the stock solution concentration was increased from before, cell damage did not occur. In the test shown in Table 5, 24-fold dilution refers to a concentration that was lower than that of No. 6 test tube diluted by 32 times with 2-fold serial dilution even if it did not return to No. 5 (16 times). that was effective in 24-fold dilution of the, in the shorthand notation can be represented as 5~6T _0. Similarly, 192
The doubling dilution had no effect on the 256-fold No. 9 test tube,
No.8 even without going back to (128-fold) that the was more potent in the middle, represented as 8~9T _0, 3 fold dilutions are No.3 (4
Fold dilution) at no effects, that it was intermediate in effect with the No.2 (2 times), it can be represented as 2~3T _0. Thus, the superiority in anti-influenza virus properties of Samples 5 and 6 in Table 4 above was further supported.

Next, among the chemical substances of the present invention, guaiacol (sample 1), ligninsulfonic acid (sample 2),
For the four types of 6-dimethoxyphenol (sample 3) and 3,5-dimethoxyphenol (sample 4), tests for evaluating antibacterial properties were performed by the following methods. (Other specimens 5, 6, 1
Test methods: Agar plates are prepared by adding 10% of each sample to a sensitive disk medium. Table 6 below shows this plate.
Test bacterial solutions (prepared at 106 cells / ml) of the ^six strains shown in
After inoculating each ml, and culturing at 37 ° C. for 48 hours, the presence or absence of growth of each bacterium is confirmed. The results are shown in Table 6 below. In the table,
(-) Indicates no bacterial growth, and (+) indicates bacterial growth.

[0044]

[Table 6] Antibacterial test results (1) Number of bacterial strains (number / ml) 1 sample 2 samples 3 samples 4 E. coli O-157 2.4 × 10 ⁶ (-) (-) (-) (-) (E .coli O157) Klebsiella pneumoniae 3.1 × 10 ⁶ (-) (-) (-) (-) (K. pneumoniae) Enteritidis 1.9 × 10 ⁶ (-) (-) (-) (-) (-.) (S. Enteritidis) Pseudomonas aeruginosa 4.2 × 10 ⁶ (-) (-) (-) (-) (P. aerginosa) Staphylococcus aureus 2.7 × 10 ⁶ (-) (-) (-) (-) (S. aureus) Bacillus subtilis 6.1 × 10 ⁶ (−) (−) (−) (−) (B. subtilis) From Table 6, guaiacol (sample 1), ligninsulfonic acid (sample 2), 2,6-dimethoxyphenol (sample 3) , And 3,5-dimethoxyphenol (sample 4)
Has a growth inhibitory effect on all test strains. In addition, the guaiac fat described above showed a bacterial growth inhibitory effect almost equal to or slightly better than that of guaiacol in the same antibacterial test.

Next, the remaining samples, ie, lignosulfonic acid sodium salt (sample 5), lignosulfonic acid sodium salt acetate (sample 6), lignin organosolv propionate (sample 11), 4-benzyloxyguaiacylglycerol -Β-guaiacyl ether (specimen 1
2) For syringaldehyde (sample 13) and veratryl alcohol (sample 14), the same six bacteria were evaluated for their inhibitory effect by a different test method. Test method: Prepare an 8% aqueous solution of each sample (powder), dilute it 40-fold and serially dilute (40- to 400-fold) the sample stock solution for MIC measurement. After inoculating 6 kinds of test bacteria inoculated into the agar medium and culturing at 30 ° C. for 17 hours, the growth inhibition zone on the agar medium was observed and judged. The results are shown in Table 7 below. The stock solution was obtained by diluting 25 ml of an 8% aqueous solution with 1000 cc of distilled water to make it 40 times, and it was 2 mg / ml in terms of solid content.

[0046]

[Table 7] Antibacterial test results (2) Strains 5 specimens 6 specimens 11 specimens 12 specimens 13 specimens 14 E. coli O-157 <40 times <40 times <40 times <40 times <40 times <40 times <40 times (E. E. coli O157) K. pneumoniae <40 times <40 times <40 times <40 times <40 times <40 times (K.pneumoniae) Enteritidis <40 times <40 times <40 times <40 times <40 times <40 times <40 times (S Enteritidis) P. aerginosa <40 times <40 times <40 times <40 times <40 times <40 times <40 times < 40 times <40 times <40 times <40 times <40 times <40 times <40 times (S. aureus) Bacillus subtilis <40-fold <40-fold <40-fold <40-fold <40-fold <40-fold (B. subtilis) In Table 7, <40-fold indicates that growth was completely inhibited by a 40-fold dilution. And a concentration of 40 times or less (for example, 30 times = 3
mg / ml, or 20-fold = 4 mg / ml). An example in which growth was completely inhibited by 40-fold dilution (2 mg / ml) is against staphylococcus aureus of lignin organosolv (sample 11). However, other examples of <40-fold (less than 40-fold) also showed partial inhibition, and it is expected that complete inhibition would be sufficiently realized by increasing the concentration. In that sense, all of the samples 5, 6, 11 to 14 are determined to have sufficiently useful antibacterial properties.

As mentioned above, the antiviral and antibacterial chemical substances identified by the present invention are all used alone in the disclosed test examples, but 3,5-dimethoxyphenol is used in combination with other agents before. As described above, not only the specimens 1 to 14 may be used alone, but also two or more specimens may be mixed or used in combination, and if necessary, one or more specimens other than the product of the present invention may be used. Can also be used in combination with the above substances. As a result, the antiviral and antibacterial properties of each substance are not diminished,
Rather, they have the effect of enhancing or reducing cell damage.

The substances having excellent antiviral and antibacterial properties according to the present invention include antiviral agents or antibacterial agents, or broadly as antimicrobial agents, as well as health foods and health drinks having multiple effects. It can also be implemented in the form of The safety when taking orally taking (or ingesting) is, of course, safe if a cell-level test confirms zero damage, such as lignosulfonic acid sodium salt (sample 5) or acetate (6). Yes (these manufacturers have also stated that they are non-toxic), but substances that show relatively strong cytotoxicity, such as 2,6-dimethoxyphenol, can be treated as safe based on previous knowledge .
According to conventionally known technical literature, 2,6-dimethoxyphenol, together with guaiacol, syringaldehyde, etc., has an LD ₅₀ value of 10 or _{less when} administered intraperitoneally or orally to mice.
It is described as being greater than 00 mg / kg and excellent in safety. In addition, it is described in the literature that when humans take guaiacol for internal use or external use, the usual dose is 0.2 g once a day and 0.6 g per day.
It is also specified that HMN is 43 mg / kg. Thus, the knowledge of the state of the art today indicates that there is no problem in the safety of all substances according to the invention.

As described in detail above, the present invention selects a wide range of candidates for a large number of industrially produced chemicals and evaluates them for antiviral (AIDS and influenza) and antibacterial properties in various aspects. Evaluated and confirmed, the 14 chemicals disclosed according to the present invention selected from among them not only exert excellent effects not previously expected for specific individual targets such as viruses and bacteria, but also It has established a new position as a versatile antimicrobial agent that exerts versatile effects including antiviral to antibacterial properties. This will provide society with a relatively low cost, stable and continuous effective antimicrobial agent, and add it to foods and beverages and take it daily for human health and well-being. Practical effects that can make a significant contribution to are achieved.

[Brief description of the drawings]

FIG. 1 is a graph showing a molecular weight distribution of a reagent used for molecular weight measurement in the present invention.

FIG. 2 is a graph showing the molecular weight distribution of ligninsulfonic acid, one of the antimicrobial agents disclosed in the present invention.

FIG. 3 is a graph showing the molecular weight distribution of the lignosulfonic acid sodium salt according to the present invention.

FIG. 4 is a graph showing the molecular weight distribution of lignosulfonic acid sodium salt acetate according to the present invention.

FIG. 5 is a graph showing the molecular weight distribution of the lignin organosolv propionate according to the present invention.

FIG. 6 is a graph showing the molecular weight distribution of humic acid examined in connection with the present invention.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (Reference) A61K 31/185 A61K 31/185 31/22 31/22

Claims (8)

[Claims] Guaiacol, lignin sulfonic acid,
2,6-dimethoxyphenol, 3,5-dimethoxyphenol, lignosulfonic acid sodium salt, lignosulfonic acid sodium salt acetate, lignin organosolve propionate, 4-benzyloxyguaiacylglycerol-β-guaiacylether, An antimicrobial agent comprising one or more active ingredients having an antimicrobial activity selected from ringaraldehyde and veratryl alcohol. 2. The method according to claim 1, wherein the active ingredient is guaiacol, ligninsulfonic acid, 2,6-dimethoxyphenol, 3,5.
-Selected from dimethoxyphenol, lignosulfonic acid sodium salt, lignosulfonic acid sodium salt acetate, lignin organosolv propionate, 4-benzyloxyguaiacylglycerol-β-guaiacylether, syringaldehyde and veratryl alcohol The antimicrobial agent according to claim 1, wherein the antimicrobial activity is antiviral activity and antibacterial activity. 3. The active ingredient comprises guaiacol, ligninsulfonic acid, 2,6-dimethoxyphenol, 3,5
-Dimethoxyphenol, lignosulfonic acid sodium salt, lignosulfonic acid sodium salt acetate, lignin organosolv propionate, 4-benzyloxyguaiacyl glycerol-β-guaiacyl ether, syringaldehyde and veratryl alcohol 2. The antimicrobial agent according to claim 1, wherein the antimicrobial activity is antiviral activity and antibacterial activity. 4. The antiviral activity is anti-AIDS virus activity and anti-influenza activity, and the antibacterial activity is against Escherichia coli O-157, Klebsiella pneumoniae, Enteritidis, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis. The antimicrobial agent according to claim 2 or 3, which has antibacterial activity. 5. The antimicrobial agent according to claim 2, wherein the active ingredient is lignosulfonic acid sodium salt or lignosulfonic acid sodium salt acetate, and has particularly remarkable anti-AIDS virus activity. 6. The antimicrobial agent according to claim 2, wherein the active ingredient is lignin organosolv propionate and has excellent antibacterial activity. 7. The antimicrobial agent according to claim 2, wherein the active ingredient is syringaldehyde, and the antimicrobial activity is anti-AIDS virus activity and anti-influenza virus activity. 8. The antimicrobial agent according to claim 2, wherein the active ingredient is guaiac oil containing guaiacol, and the antimicrobial activity is an antiviral activity.