JPH11246319A - Antimicrobial and antifungal agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antimicrobial and antifungal agent that can largely alleviate the irritating smell and can be applied to various kinds of beverage and food by using a specific ω-alkylsulfinylalkykl isothiocyanate as an active ingredient. SOLUTION: This antimicrobial and antifungal agent comprises an ω- alkylsulfinylalkyl isothiocyanate represented by the formula: R-S(O)-(CH2 )n -NCS (n is 1-10; R is a 1-4C alkyl). In an embodiment, this isothiocyanate is 3- methylsulfinylpropyl isothiocyanate, 6-methylsulfinyl-propyl isothiocyanate or the like. The compound in formula I where n is <=7 is included in the flavor components of horse radish, but its content is low. The compound of the formula is prepared by oxidizing ω-methylthioalkyl isothiocyanate of the formula: CH3 S-(CH2 )n -NCS with a peroxide.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an antibacterial antifungal agent comprising an ω-alkylsulfinylalkylisothiocyanate compound. By mixing the antibacterial antifungal agent of the present invention into various foods and drinks or oral hygiene agents such as toothpastes, mouthwashes and breath fresheners (hereinafter collectively referred to as foods and drinks), various fungi such as bacteria and fungi can be obtained. Etc. can be effectively inhibited.

[0002]

2. Description of the Related Art It has long been known that extracts of cruciferous plants such as wasabi and mustard have a growth inhibiting and bactericidal action against microorganisms. Among them, allyl isothiocyanate (CH ₂ ＣＨCHCH ₂ NC) which is a main component in an aroma extract
S) is an antibacterial agent against Escherichia coli and Staphylococcus aureus, which are the causative agents of food poisoning, and has been studied and practically used as a freshness preserving agent for fish meat and animal meat from the viewpoint of safety for the human body. I have.

However, allyl isothiocyanate has a high volatility, so that it is necessary to maintain a constant volatilization at an appropriate concentration for a long period of time, and it is necessary to devise a formulation such as microencapsulation. Since it has an irritating odor (detection threshold: 0.046 ppm), it has a drawback that the original taste and aroma of the added food and drink are impaired and versatility is poor.

On the other hand, regarding the antibacterial properties of isothiocyanate compounds other than allyl isothiocyanate, Salvador
a persica L. That benzyl isothiocyanate extracted from bacterium is antibacterial against Streptococcus mutans, a causative fungus,
In addition, it has been reported that benzyl isothiocyanate and 4-methoxybenzyl isothiocyanate extracted from the roots of Hippocrata welwitschii have antibacterial properties against common bacteria, but, for example, that the pungent odor is not improved At present, problems such as these remain, and they have not yet been put to practical use.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to eliminate the above-mentioned drawbacks in the prior art, to greatly reduce the irritating odor, to be safe and widely applicable to various foods and drinks, and to obtain various kinds of bacteria. An object of the present invention is to provide an antibacterial antifungal agent excellent in inhibiting the growth of fungi. In particular, we provide an excellent antibacterial antifungal agent for foods and drinks with a high threshold value, which has the same antibacterial antifungal activity as allyl isothiocyanate, which has been widely used in spite of the conventional problems, and has greatly reduced irritation odor. It is to be.

[0006]

Means for Solving the Problems The present inventors have newly found that the ω-alkylsulfinylalkylisothiocyanate compound contained in wasabi and horseradish (horseradish) is excellent as an antibacterial and antifungal agent. The invention has been completed. That is, the present invention has the general formula (I) R-S (O ) - (CH 2) n -NCS (I) ( wherein, n represents an integer of 1 to 10, R is alkyl of C ₁ -C ₄ Which represents an alkylsulfinylalkylisothiocyanate compound having the following formula: Further, the present invention is a food and drink and an oral hygiene agent containing the above antibacterial antifungal agent.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail. (1) ω-alkylsulfinylalkylisothiocyanate compound In the general formula: R—S (O) — (CH ₂ ) _n —NCS (I), n represents an integer of 1 to 10, but 2 to 8, especially 3
To 7, especially 3 to 6 are preferred. R represents a C ₁ -C ₄ alkyl group, and preferred examples include methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl.

Examples of the compound having the formula (I) include methylsulfinylmethylisothiocyanate, ethylsulfinylmethylisothiocyanate, n-propylsulfinylmethylisothiocyanate, isopropylsulfinylmethylisothiocyanate, and n-butylsulfinylmethyl Isothiocyanate, 2-methylsulfinylethyl isothiocyanate, 2-ethylsulfinylethyl isothiocyanate, 2-n-propylsulfinylethyl isothiocyanate, 3-methylsulfinylpropyl isothiocyanate, 4-methylsulfinylbutyl isothiocyanate , 5-methylsulfinylpentylisothiocyanate, 6-methylsulfinylhexylisothiocyanate, 7-methylsulfinylheptylisothiocyanate Cyanate, 8-methylsulfinyl-octyl isothiocyanate, etc. 9 methylsulfinyl nonyl isothiocyanate and the like.

In the above formula (I), it is known that compounds having n of 7 or less are contained in aroma components such as wasabi and horseradish. In order to obtain a target, it is appropriate to use synthesis. When R is methyl, the compound having the above formula (I) can be prepared, for example, according to the method described in JP-A-7-247497 in an inert solvent in the following formula (II): CH ₃ S— (CH ₂ ) _n − It can be produced by oxidizing ω-methylthioalkylisothiocyanate represented by NCS (II) (wherein n has the same meaning as described above) with a peroxide.

In the above process, the inert solvent is not particularly limited, but methylene chloride, chloroform and the like are preferable, and as the peroxide, m-chloroperbenzoic acid, hydrogen peroxide, peracetic acid and the like are preferably used. Such a peroxide is used in a proportion of 1 to 1.5 equivalents, preferably 1 to 1.1 equivalents, based on the compound of the formula (II), and the reaction is carried out at -10 to 2
It is carried out at a temperature of 0 ° C, preferably -5 to 5 ° C for 1 to 5 hours. After completion of the reaction, the reaction mixture is purified by distillation or silica gel chromatography to obtain an ω-methylsulfinylalkylisothiocyanate compound.

(2) Application of antibacterial and antifungal agents The antibacterial and antifungal agents of the present invention are incorporated into foods and drinks at a concentration of preferably 1 to 100 ppm, particularly preferably 10 to 50 ppm, according to a conventional method. If it is less than 1 ppm, the antibacterial and antifungal effect is not sufficiently exhibited, while if it exceeds 100 ppm, its own odor tends to impair the taste and aroma of foods and drinks.

The oral hygiene preparation containing the antibacterial and antifungal agent of the present invention may be in the form of a liquid, a solid, or a semi-solid, a dentifrice, a troche, a liquid or powdery mouthwash. It is applied to oral solution, bad breath inhibitor, chewing gum and the like. In addition, foods and beverages containing the antibacterial antifungal agent of the present invention include fishery products such as kamaboko, chikuwa, hampan, fish meat sausage / ham, surimi products, dried seafood / smoked products, etc .; Semi-solid marine products such as taro, mustard, etc .; livestock meat products such as ham, (winner) sausage, bacon, minced meat; salads, hamburgers, gyoza, shumai, tsukudani, boiled beans, etc .; Chinese cabbage, cucumbers, vegetables Pickles such as shallow pickles, kimchi, zasai, takuan, mustard lotus root, etc .; Seasoning liquids such as sauce and soup; Seasonings such as miso, soy sauce; Noodles such as raw or boiled soba, udon, pasta; Vegetables such as vegetables and fruits, semi-cooked products of these; other semi-processed frozen foods; fruit juice, carbonated drinks, tea, milk drinks, lactic acid drinks,
Various drinks such as coffee, cocoa, soy milk; condensed milk, powdered milk, fermented milk, butter, cheese, ice cream, fresh cream and other dairy products; caramel, candy, chewing gum,
Jam, margarine and the like.

Further, the antibacterial antifungal agent of the present invention can be used for various products called so-called antibacterial goods (products), for example,
Kitchen utensils such as cutting boards; toiletries such as toothbrushes; stationery such as writing utensils, erasers and notebooks; clothing such as underwear; car interiors such as handles and sheets; home appliances such as word processors and refrigerators; It can be widely applied to interior materials and the like. That is, the antibacterial and antifungal agent is kneaded and mixed with the raw materials in the production stage of the product, or the agent is liquefied by adding an auxiliary agent such as a surfactant as necessary using a solvent to the surface of the product. It is applied by spraying or spraying. In addition, the liquid or sprayed this agent,
It can be applied as a disinfectant or fungicide for kitchen toilets, toilet seats and bathroom dill. In addition, the liquid preparation of the present invention can be impregnated into a liquid-absorbing support such as paper or cloth to be applied as sanitary cleaners or hygiene articles such as wet tissues.

The antibacterial and antifungal agent of the present invention can be used in combination with other antibacterial agents such as alcohol; spice components such as sage and rosemary; and organic acids such as citric acid, lactic acid and acetic acid.

Next, the minimum inhibitory concentrations (MI
The effect of the antibacterial antifungal agent of the present invention is shown by measuring C). (1) Staphylococcus aureus IFO 12
Antimicrobial test against 732) During culture of the bacterium, a medium A having the following composition was used. (Medium A) Peptone 5.0 g Yeast extract 1.5 g Meat extract 1.5 g Sodium chloride 3.5 g Glucose 1.0 g Disodium hydrogen phosphate 3.0 g Potassium dihydrogen phosphate 1.32 g Distilled water 1000 ml

The test bacteria were inoculated into the medium A and cultured with shaking at 35 ° C. for 12 hours. Then, 1 × 10 ⁶ c
It was adjusted to be ells / ml. 324 μl of the bacterial solution was placed in the first row of a 96-well microplate, and 180 µl was placed in the second to 11th rows.
μl of each of the isothiocyanate solutions 3 shown in Table 1.
6 μl was added to the first row. After stirring, the first row of solution 18
Transfer 0 ml to the second row and make a serial two-fold dilution series,
The culture was allowed to stand at a temperature of 6 ° C. for 6 hours. The growth of the bacteria in each well was determined with a microplate reader (when the absorbance (optical density) at a wavelength of 630 nm was 0.05 or less) and the naked eye.
Table 1 shows the minimum inhibitory concentrations of various isothiocyanates.

(2) Escherichia coli IFO 330
Antibacterial test for 1) The test was performed in the same manner as that for Staphylococcus aureus described above. Table 1 shows the minimum inhibitory concentrations of various isothiocyanates.

(3) Pseudomonas aeruginosa I
FO 3080) An antibacterial test was performed in the same manner as described above for Staphylococcus aureus. Table 1 shows the minimum inhibitory concentrations of various isothiocyanates.

(4) Bacillus subtilis IFO 31
Antibacterial test against 34) The test bacteria were inoculated into medium A and cultured with shaking at 35 ° C for 12 hours. Then, the spores in this bacterial solution were 1 × 10 ⁶ cells
/ Ml. 324 μl of the bacterial solution was placed in the first row of the 96-well microplate, and 180 μl was placed in the second to eleventh rows, and 36 μl of the various isothiocyanate solutions shown in Table 1
Was added to the first row. After agitation, 180 ml of the solution in the first row was transferred to the second row to form a serial two-fold dilution series, which was incubated at 35 ° C for 6 hours. The growth of the bacteria in each well was determined with a microplate reader (when the absorbance (optical density) at a wavelength of 630 nm was 0.05 or less) and the naked eye. Table 1 shows the minimum inhibitory concentrations of various isothiocyanates.

(5) Black mold (Aspergillus niger IFO)
Antifungal test against 4414) During culture, a medium B having the following composition was used. (Medium B) Peptone 10.0 g Glucose 40.0 g Distilled water 1000 ml

The test bacteria were inoculated into the medium B and cultured with shaking at a temperature of 25 ° C. for 24 hours. Then the spores in this bacterial solution were
It was adjusted to be × 10 ⁴ cells / ml. 324 μl of the bacterial solution was added to the first row of the 96-well microplate, and 180 μl to the second to eleventh rows, and 36 μl of the various isothiocyanate solutions shown in Table 1 were added to the first row. After agitation, 180 ml of the solution in the first row was transferred to the second row to form a serial two-fold dilution series, which was incubated at 25 ° C. for 15 hours. Germination of spores in each well was determined microscopically. Table 1 shows the minimum inhibitory concentrations of various isothiocyanates.

(6) Candida albicans IFO
Antibacterial test against 1835) A test bacterium was inoculated into the above-mentioned medium B and cultured with shaking at a temperature of 25 ° C. for 24 hours. Then, this bacterial solution was added to 1 × 10 ⁵ cells / ml.
It was adjusted to be. 324 μl of the bacterial solution was placed in the first row of a 96-well microplate, and 180 μl was placed in the second to 11th rows.
36 μl of the various isothiocyanate solutions described in Table 1 were added to the first row. After stirring, add 180 ml of the solution in the first row to the second
The cells were transferred to a column, and a serial two-fold dilution series was prepared, followed by stationary culture at a temperature of 25 ° C. for 22 hours. The growth of the bacteria in each well was determined with a microplate reader (when the absorbance (optical density) at a wavelength of 630 nm was 0.05 or less) and the naked eye. Table 1 shows the minimum inhibitory concentrations of various isothiocyanates.

[0023]

[Table 1]

Next, the threshold values of various isothiocyanates were measured as follows. For the threshold measurement, the 2: 5 point method was used according to the following Amoore's method. 20 ml of water was placed in five brown glass cups, two of which were diluted with an aqueous solution 2 diluted to a concentration at which the evaluator could detect isothiocyanate in the sample.
Add 0 ml. Water is used after removing residual chlorine by passing through a filter ("D-4" manufactured by Organo) equipped with activated carbon. The temperature of the aqueous solution is 20 ± 1 ° C, the room temperature is 2
The temperature is set at 0 ± 5 ° C. and the humidity is set at 60 to 80%, and the measurement is performed in the morning. The evaluators consisted of 10 males in their twenties, and each evaluator selected two cups that contained the water or sample solution in the five cups little by mouth and felt that the sample was contained. If this selection is correct, the same sensory evaluation is performed with the sample concentration further reduced by 10 ^-0.5 times, and the process is repeated until a wrong answer is given. The geometric mean value of the sample concentration at the time when each evaluator finally gives the correct answer is set as the threshold. The results of the threshold test were as shown in FIG.

The results of the above antibacterial test and threshold test are summarized as follows. It has been found that the ω-methylsulfinylalkylisothiocyanate of the present invention wherein R is methyl in the formula (I) has a strong antibacterial effect against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and black mold. In particular, 4-methylsulfinylbutyl isothiocyanate was found to outperform allyl isothiocyanate's antibacterial activity against Staphylococcus aureus, Bacillus subtilis and Escherichia coli. Further, ω-methylsulfinylalkyl isothiocyanate has a higher threshold value than ω-alkenyl isothiocyanate such as allyl isothiocyanate, which is another isothiocyanate compound, and ω-methylthioisothiocyanate. It was also found that when the same amount was added, the original flavor of food and drink was not impaired.

[0026]

EXAMPLES Next, the present invention will be described more specifically with reference to Reference Examples and Examples. Synthesis Example of ω-alkylsulfinylalkylisothiocyanate Reference Example 1 Production of 3-methylsulfinylpropylisothiocyanate 12.8 g of 3-methylthiopropylisothiocyanate
Is dissolved in 200 g of methylene chloride and cooled to -5 ° C.
15.6 g of m-chloroperbenzoic acid in methylene chloride (32
0 g) The solution is added dropwise and stirred for 1 hour. The reaction solution is washed with a saturated aqueous solution of sodium bicarbonate and a saturated saline solution and then separated to obtain an organic layer. Then, after concentration under reduced pressure, purification by silica gel chromatography gave 12.1 g (yield 85%) of 3-methylsulfinylpropyl isothiocyanate.

Reference Example 2 Production of 6-methylsulfinylhexyl isothiocyanate 14.4 g of 6-methylthiohexyl isothiocyanate
Is dissolved in 225 g of methylene chloride and cooled to -5 ° C.
14.8 g of m-chloroperbenzoic acid in methylene chloride (34
0 g) The solution is added dropwise and stirred for 3 hours. The reaction solution is washed with a saturated aqueous solution of sodium bicarbonate and a saturated saline solution and then separated to obtain an organic layer. Then, after concentration under reduced pressure, purification by silica gel chromatography gave 13.8 g (88% yield) of 6-methylsulfinylhexylisothiocyanate.

Example 1 Preparation of candy A candy containing the antibacterial antifungal agent of the present invention was prepared by a conventional method using a candy composition comprising the following components. Sugar 50 starch syrup 33 organic acid 2 flavor 0.2 water balance 6-methylsulfinylhexyl isothiocyanate 0.002 total 100g

Example 2 Preparation of chewing gum Chewing gum containing the antibacterial antifungal agent of the present invention was prepared by a conventional method using a chewing gum composition comprising the following components. Gum base 20.09 sugar 53 glucose 10 starch syrup 16.4 fragrance 0.5 5-methylsulfinylpentyl isothiocyanate 0.01 total 100g

Example 3 Toothpaste A toothpaste containing the antibacterial antifungal agent of the present invention was prepared by a conventional method using a toothpaste composition comprising the following components. Calcium hydrogen phosphate 20 Silicic anhydride 10 Sorbit liquid 40 Glycerin 10 Benzalkonium chloride 0.01 Thickener 0.6 Fragrance 0.8 Water Remainder 6-methylsulfinylhexyl isothiocyanate 0.002 Total 100 g

Example 4 Mouthwash (Mouthwash) A mouthwash containing the antibacterial antifungal agent of the present invention was prepared by a conventional method using a mouthwash composition comprising the following components. Ethyl alcohol 10.0 Polyoxyethylene hydrogenated castor oil 2.0 Saccharin sodium 0.2 Sodium benzoate 0.1 Glycerin 5.0 Fragrance 0.2 Sodium citrate (pH adjuster) Water Water balance 4-Methylsulfinylbutyl isothiocyanate 0.002 total 100g

[0032]

Industrial Applicability The antibacterial antifungal agent of the present invention can effectively inhibit the growth of fungi such as various kinds of bacteria and mold which are frequently in contact with daily life, and is a component derived from edible plants such as wasabi and mustard. Therefore, it has high safety to the human body and is suitable for addition to foods and drinks and oral hygiene agents. Also, compared to other isothiocyanate compounds such as allyl isothiocyanate which have been conventionally used, the irritating odor is remarkably reduced and the threshold value is high, so even if added at a high concentration, the original taste of food and drink etc. No odor or smell. Therefore, it can be added in a wide range of low to high concentrations depending on the type of the bacterium to be treated, and is excellent in practicality. Furthermore, since it has low volatility, it is excellent in handleability in compounding.

[Brief description of the drawings]

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1: The threshold value of the ω-methylsulfinylalkyl isothiocyanate of the present invention when R is a methyl group in the general formula: RS (O) — (CH ₂ ) _n -NCS and other isothiocyanate systems 3 is a graph showing a comparison with a threshold value of a compound. Other compared isothiocyanate compounds are ω-methylthioalkylisothiocyanates represented by the general formula CH ₃ —S— (CH ₂ ) _n —NCS, and the general formula CH ₂ ＣC
Ω-Alkenyl isothiocyanate represented by H- (CH ₂ ) _n-2- NCS. The vertical axis represents the threshold value (unit: ppb), and the horizontal axis represents n in each of the above equations.

Claims (3)

[Claims] 1. A compound represented by the general formula (I): RS (O)-(CH ₂ ) _n -NCS (I) wherein n is an integer of 1 to 10, and R is a C _{1 to} C ₄ alkyl. Which represents an ω-alkylsulfinylalkylisothiocyanate compound having the formula: 2. A food or drink comprising the antibacterial antifungal agent according to claim 1. 3. An oral hygiene agent comprising the antibacterial antifungal agent according to claim 1.