JP2002212090A - Antibacterial agent for methicillin-resistant staphylococcus aureus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antibacterial agent which exhibits an extremely remarkable antibacterial activity against staphylococcus, especially Methicillin-resistant Staphylococcus aureus. SOLUTION: This antibacterial agent contains a hot water extract obtained by extracting the leaves and/or stems of SENRINKEI alpine tea and/or HAKUGO oolong tea produced in Taiwan as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a staphylococcus (Staphyloco
ccus sp.), a methicillin-resistant Staphylococcus aureus (methicillin resi
stant Staphylococcus aureus (hereinafter referred to as MRSA)
The present invention relates to an antibacterial agent which exerts an antibacterial action on the antibacterial agent.

[0002]

BACKGROUND OF THE INVENTION Staphylococci are known as pathogenic bacteria of various inflammations in humans and animals, such as dermatitis, conjunctivitis, sinusitis, otitis media, cystitis, pneumonia, osteomyelitis, arthritis and visceral abscess. . In particular, Staphylo
coccus aureus) is an important causative agent of purulence in humans and animals, and also a causative agent of food poisoning due to enterotoxin enterotoxin produced by it. In recent years, in particular, serious enteritis due to nosocomial infection or bacterial alternation due to MRSA showing resistance to various antibiotics has become a problem.

[0003] As the name implies, MRSA is Staphylococcus aureus resistant to the antibiotic methicillin. That is, MRSA has acquired resistance to a methicillin-based drug (for example, penicillin or cephalosporin drug) having a strong antibacterial action against Staphylococcus aureus.

[0004] MRSA is all penicillins; all cephem drugs called first, second and third generation;
Monobactams: Exhibit a wide range of drug resistance to all commercially available β-lactam drugs such as carbapenems. Furthermore, it is known that it also exhibits resistance to new quinolones. Recently, it has been reported that vancomycin, which has been used for the treatment of infections caused by MRSA, has also appeared to be resistant to MRSA. For example, it has been increasingly desired to provide an antibacterial agent effective for MRSA.

[0005] Furthermore, MRSA not only shows multidrug resistance, but also may exhibit extremely strong pathogenicity. MRSA especially for compromised patients with reduced resistance to infection
Infection is a serious problem. For example, it is extremely difficult to cure MRSA when serious patients such as patients suffering from immunodeficiency due to leukemia or cancer, severely burned patients, bedridden elderly people and premature babies are infected with MRSA.

[0006]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and an object of the present invention is to provide an antibacterial agent which exhibits an effective antibacterial action against MRSA.

[0007]

The methicillin-resistant Staphylococcus aureus antibacterial agent of the present invention, which has achieved the above objects, has, as an active ingredient, Suginrinkei Takayama tea native to Taiwan and / or tea leaves and / or white oolong oolong tea. Alternatively, it has a gist in containing a hot water extract extracted from a stem.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The present inventors have studied diligently with a particular focus on tea in order to provide an antibacterial agent capable of exhibiting an effective antibacterial action against staphylococci, especially MRSA.
Specifically, 35 tea leaves native to Japan, 1 tea leaf native to Taiwan
Using five kinds of tea leaves and five kinds of tea leaves native to China, the antibacterial activity against various staphylococci including MRSA was examined in detail. As a result, they have found that two types of tea leaves and / or stems of Suginrinkei Takayama tea and white oolong, native to Taiwan, can extremely effectively inhibit the growth of staphylococci, especially MRSA, and completed the present invention. .

[0009] In general, tea is prepared by collecting young shoots (leaves) of tea plant belonging to the family Camellia.
It is roughly classified into unfermented tea (green tea), semi-fermented tea (oolong tea, etc.), and fermented tea (black tea). Furthermore, for example, semi-fermented tea is roughly classified into oolong tea and wrapping tea according to the production method, the type and shape of the raw materials, and the like.

[0010] Because of its elegant aroma and taste, tea is known not only to be a favorite beverage but also to have various physiological effects. For example, it is known that catechin (tannin) contained in tea leaves in a large amount has an anticancer effect, an antiaging effect, a blood cholesterol lowering effect, and the like.

According to recent studies, the usefulness of catechins contained in tea has been actively promoted. For example, a Japanese tea leaching solution can kill highly virulent pathogenic Escherichia coli O157 and detoxify toxin. It has been reported that it has an action to do. The present inventors have also paid attention to catechin and examined whether it has an antibacterial action against staphylococci, particularly MRSA, but found that a high concentration of catechin alone does not provide a desired antibacterial effect. .

Further, various teas were examined while paying attention to the content of catechin, but no significant correlation was found between catechin and staphylococcal antibacterial activity. For example, Japanese green tea is drunk by pouring relatively low-temperature hot water, so that a relatively large amount of catechin or the like, which is a bitter component, elutes in the tea. Roasted tea is roasted sencha, but roasting reduces catechins. In addition, oolong tea,
Semi-fermented tea of wrapping tea is poured after boiling boiling water and steamed for 3 to 5 minutes, so that catechins are reduced to less than half of green tea. When the antibacterial activity against MRSA of various teas having different catechin contents was examined as described above, even if the oolong tea was considered to have a small amount of catechin as described above, the tea leaves of the white Australian oolong from Taiwan. It was found that when drinking, excellent antibacterial action was obtained. On the other hand, the antifreeze effect against MRSA was not obtained in the frozen oolong from Taiwan. On the other hand, Suginrinkei Takayama tea from Taiwan is used as oolong tea, but this is also the same as the above-mentioned white oolong oolong,
It was found to show excellent antibacterial action. Therefore, it can be seen that the antibacterial action according to the present invention is not necessarily due to the action of catechin alone.

By the way, when the amount of catechin was examined for the two kinds of tea leaves used in the present invention, it was found that the amount was very small or hardly detectable. For example, in the case of the white oolong, 15 g of this tea leaf and / or stem was immersed in 650 mL of water at 90 ° C., left at room temperature for 30 seconds, and the amount of catechin in the filtered filtrate was measured by the FOLIN-DENIS method. And catechin (detected as tannic acid) were not detected (detection limit: 0.01 g / 100 g). On the other hand, the measurement of Sugibayashi Takayama tea by the same method revealed that only 0.03 g / 100 g of tannic acid was detected.

It is unknown in detail why the above-mentioned Taiwanese tea leaves used in the present invention can exert an extremely excellent antibacterial action against MRSA. Considering that catechin alone, which was considered to be a medicinal component of catechin, does not show any antibacterial activity against staphylococci such as MRSA, Taiwan's soil, climate, cultivation conditions, and active ingredients in tea leaves are intricately intertwined. Thus, it is considered that the desired antibacterial action was exhibited.

Here, the tea used in the present invention will be described.

First, white oolong is produced only in Taiwan [Manufacturer: Xinhua Tea Co., Ltd.] The fermentation degree is about 60-7.
It is 0% blue tea (oolong tea), and is known as a type of oolong tea similar to black tea. Of the semi-fermented teas, the only one produced from tea buds is the white oolong dragon, which is characterized by a higher degree of fermentation than ordinary semi-fermented tea. The tea buds preferably have an enlarged heart bud, and tea trees such as Aishin Taiyu, Taicha No. 17, and white hair monkey are mainly used. Main production area is Hsinchu Beipu in Taiwan,
Miaoli, head, etc. In preparing the above white oolong, first, the tea leaves and / or stems are exposed to weak sunlight for a long time, and when the tea buds are slightly whitened and the epidermis of the stems shrink in a wrinkle-like manner, the tea leaves and indoors are transferred to the room. Stir until scent appears.
Next, the obtained tea buds are roasted at a low temperature, covered with a wet cloth, rubbed so as not to shatter, and then dried with hot air at about 90 ° C.

[0017] Suginrinkei Takayama tea is a green tea (oolong tea) having a fermentation degree of about 15 to 30%, which is cultivated in Suginrinkei in the Central Taiwan Mountains (main production areas are Chiayi County, Nantou County, etc.). Hemisphere type oolong (eg, blue heart oolong) tea tree is mainly used. The tea leaves have a bluish-green color due to the effects of alpine cultivation, have little bitterness or astringency, and are known as good-quality incense tea with a lingering scent that lasts forever. The production method is the same as that of ordinary oolong tea. After being exposed to weak sunlight, it is moved into a room, stirred and fermented, and then obtained through roasted tea, heating, kneading, and drying. Specifically, Xinhua tea line (Yes)
May be used.

The antibacterial agent of the present invention contains the above-mentioned hot water extract of tea leaves and / or stem as an active ingredient. In order to obtain a desired antibacterial effect, it is recommended to adjust the concentration of the hot water extract to about 2 to 15%, preferably 2 to 10%. As is clear from the examples described later, the higher the concentration of the hot water extract, the higher the antibacterial effect tends to increase.However, taking into account the balance with ease of taking, etc., usually, It is recommended to pour about 100 cc of hot water for about 2 to 10 g of tea leaves. For example, when used as a tea pack, pack 2.5 g or 5.0 g per pack, and add 6 g
It is preferable to pour about 100 cc of hot water at 5 to 95 ° C. for taking. It is recommended to take about 1.2 to 2.0 cc / kg of the hot water extract adjusted to such a concentration at a time, 3 to 4 times a day.

In the present invention, the above-mentioned hot water extract of the tea leaves and / or stems of Sugirinkei Takayama tea and Baiyu Oolong from Taiwan may be used alone or in combination. It does not matter. When both are used in combination, it is preferable to use approximately equal amounts so that the total content falls within the above range. Of course,
The content is not intended to be limited to this amount, and the content can be appropriately adjusted according to the condition, sex, weight, and the like of the subject within a range in which a desired antibacterial action is obtained.

Further, the above antibacterial agent may appropriately contain vitamin C, vitamin E, vitamin K, folic acid, sweetener, excipient, stabilizer and the like as long as the action of the present invention is not impaired. It is.

The antibacterial agent of the present invention is Staphylococcus, that is, Staphy
Bacteria belonging to the genus lococcus, for example, S. aureus containing MRSA, S. epidermi
dis) and saprophytic staphylococcus (S. saprophyticus), and among others, exerts an extremely strong antibacterial action on MRSA.

Hereinafter, the present invention will be described in detail based on embodiments. However, the following embodiments do not limit the present invention, and all modifications and alterations without departing from the spirit of the preceding and following embodiments are included in the technical scope of the present invention.

[0023]

EXAMPLES Example 1: Sensitivity test to MRSA using various Taiwanese teas and Chinese oolong tea The following tests were performed using various Taiwanese teas shown in Table 1 and various Chinese oolong teas shown in Table 2. Was served.

First, methicillin-resistant Staphylococcus aureus (M
RSA) was grown and cultured in a nutrient medium, and then subjected to a halo test, that is, a paper disk sensitivity test. Specifically, a disc having a diameter of 8 mm manufactured by Showa Disc was impregnated with tea of each concentration shown in Tables 1 and 2. What was picked from each bacterial colony was spread on a plate medium, and the above-mentioned three discs were set on the plate medium.
After culturing for 4 to 48 hours, the width of the growth inhibition circle was measured.
Table 1 shows the results.

[0025]

[Table 1]

[0026]

[Table 2]

From these results, of the various types of tea leaves from Taiwan and China shown in the table, even at an extremely low concentration of 2.5%, the antibacterial effect was excellent against MRSA. It can be seen that there are only the tea leaves of the white oolong oolong and the tea leaves of Sugirinkei Takayama tea.

On the other hand, other tea leaves showed almost no antibacterial effect, and showed only a slight antibacterial effect only when adjusted to an extremely high concentration of 12.5 to 15.0%. Was.

Although not shown in the table, it was confirmed by experiments that similar test results were obtained for staphylococci other than MRSA.

Example 2: Susceptibility test of Sugibayashi Takayama tea to various bacteria (1) Test bacteria Methicillin-resistant Staphylococcus aureus (MRS)
A), Staphylococcus aureus (Staphylococcus aureus wild strain and Staphylococcus aureus 209P), Staphylococcus aureus (Staphylococcus epidermidis, and Staphyloco)
ccus haemolyticus), saprophytic staphylococci (Staphylococcu)
s saprophyticus) Staphylococcus; Streptoco
ccus pneumoniae, Streptococcus agalactiae Streptococcus; Enterococcus faealis Streptococcus; and Escehr
ichia coli E. coli, enterohemohemorrhagic E. coli (O
157); Salmonella enteritidis, Salmonella thyphimu
A total of 13 strains such as Salmonella such as rium were used.

(2) Test method A susceptibility test was performed in the same manner as in Example 1 except that the above test bacteria were grown and cultured in an optimal nutrient medium common to each bacterial species in advance. Table 3 shows the results.

[0032]

[Table 3]

From Table 3, it was found that the use of the tea leaves of Sugibayashi Takayama tea exhibited an extremely excellent antibacterial effect on staphylococci, especially MRSA. Although the antibacterial activity of the same tea leaf extract is clear at a glance, the inhibition circle for MRSA was much larger than the inhibition circle for staphylococcal standard strain 209P and the wild strain.

On the other hand, antibacterial activity was hardly observed for other strains other than staphylococci, and only when the concentration was increased to a high concentration, the antibacterial activity was only slight.

For reference, MRS among staphylococci
A, Staphylococcus aureus wild strain,
And Staphylococcus aureus 209P), and a photograph showing the results of the inhibition circle for O157 are shown in FIG.

Example 3: Susceptibility test for each species of white oolong Oolong The same susceptibility test as in Example 2 was carried out except that white long oolong was used instead of Sugirinkei Takayama tea in Example 2. Was. Table 4 shows the obtained results.

[0037]

[Table 4]

From Table 4, it can be seen that even when the tea leaves of the white oolong oolong were used, an excellent antibacterial action against staphylococci was exhibited. As in the case of Sugibayashi Takayama tea, an extremely strong antibacterial action against MRSA was observed.

For reference, MRSA among staphylococci,
FIG. 2 shows photographs showing the results of inhibition circles against Staphylococcus aureus (Staphylococcus aureus wild strain and Staphylococcus aureus 209P) and O157.

Example 4: Test for Confirming Bactericidal Effect (In Vitro Test) For the purpose of finally confirming whether Sugibayashi Takayama tea and white oolong dragon used in the present invention have a bactericidal effect against staphylococci. , Add each Taiwanese tea leaf liquor to the viable bacterial suspension,
After a certain period of time, it was examined whether the bacteria were killed.

Specifically, the three types of MRS shown in Table 3
In each test tube containing 5 cc of A suspension, add Sugibayashi Takayama tea leaves /
After adding 5 cc of a 5% solution of white oolong tea leaves, the mixture was allowed to stand in an incubator at 37 ° C. for 24 hours, centrifuged, and the precipitate was collected and spread on Staphylococcus No. 110 medium. Further, the cells were cultured in the above incubator for 24 hours, and when no colony was generated, it was judged as "-".

Table 5 also shows the obtained results.

[0043]

[Table 5]

From Table 5, it was confirmed that the colonies were not observed at all when the tea leaf extract of Sugirinkei Takayama tea and Baigo oolong from Taiwan was added, and that each MRSA strain was killed. Was done.

Example 5: Test for confirming bactericidal effect (in vivo test) In order to further demonstrate the bactericidal action of the antibacterial agent of the present invention,
In addition to the in vitro test, an in vivo test using rats was performed.

In detail, a total of 24 mature SD healthy male rats derived from SPF were divided into 4 rats / group, and the three kinds of MRSA bacteria suspensions shown in Table 6 were directly injected into the stomach at 5 cc / animal using a sonde needle for rats. Was administered. After 30 minutes from the administration, 5.0% tea leaf extract from each Taiwan was similarly intragastrically administered at 5 cc / animal. After 24 to 48 hours, the bacteria were collected from the collected feces and smeared on Staphylococcus No. 110 medium.
The cells were cultured in an incubator at 24 ° C. for 24 to 48 hours. The colonies that developed were stained with Gram and examined microscopically to see if they were Staphylococcus. The results are shown in Table 6.
Before performing this experiment, it was confirmed that staphylococci were not present in the feces of the rats subjected to the experiment.

[0047]

[Table 6]

From Table 6, no colonies were observed in any of the Taiwanese tea leaf extracts, confirming that each MRSA strain was killed.

[0049]

According to the present invention, an antibacterial agent having a very remarkable antibacterial action against staphylococci, particularly MRSA, can be provided. Since the antibacterial agent of the present invention uses a hot water extract extracted from the above-mentioned two kinds of tea leaves / stalks, it is extremely useful as an antibacterial agent that can be taken safely and easily without side effects.

[Brief description of the drawings]

FIG. 1. In Example 2, MRSA, Staphylococcus aureus (Staphylococcus aureus wild strain, and Staphylococcu
21 shows photographs showing the results of inhibition circles for S aureus 209P) and O157.

FIG. 2. In Example 3, MRSA, Staphylococcus aureus (Staphylococcus aureus wild strain, and Staphylococcu
21 shows photographs showing the results of inhibition circles for S aureus 209P) and O157.

Claims (1)

[Claims] 1. A methicillin-resistant Staphylococcus aureus antibacterial agent comprising, as an active ingredient, a Taiwanese Sugirinkei Takayama tea and / or a tea extract and / or stem hot water of Bailong Oolong.