TW586931B - Antimicrobial agent for enterococcus - Google Patents

Abstract

To obtain an antimicrobial agent having extremely excellent antimicrobial action on a vancomycin-resistant enterococcus among enterococci by making the antimicrobial agent include an extract of the cap pileus of fruit body of Ganoderma lucidum. This antimicrobial agent contains an extract of cap pileus part of fruit body of Ganoderma lucidum, preferably ripened into full maturity, containing a large amount of spores. Preferably the agent further contains royal jelly. The extract is obtained by using Ganoderma lucidum having about 14-17 cm diameter of cap pileus, >= about 35 cm thickness of cap pileus, a size of about 100 g of two of the mushrooms calculated as dry weight except stems, thinly cutting the cap pileus parts, extraction-treating the cut cap pileus parts with hot water at 95-100 DEG C, filtering a treated solution having reached the highest concentration and spray-drying.

Description

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 586931 A7 _ Η " 7 V. Description of the invention (0 [Technical Field of the Invention] The present invention relates to an antibacterial agent having an effect against enterococci. The antibacterial agent of the present invention is effective against enterococci It is effective for various inflammation symptoms caused by inflammatory bacteria. In addition, for patients with pyogenic symptoms such as Staphylococcus aureus and Streptococcus pyogenes, which spread throughout the body through blood circulation, and patients with severe enteritis or People with significantly lower physical strength and patients with debilitating conditions also have an effect. The use of the antibacterial agent of the present invention can exert the antibacterial effect of VRE: Vancomycin Resistant Enterococcus, so it is extremely useful. [Knowledge technology Enterococci are streptococci that are often present in the intestine with group D antigens classified in the Lancefield serum, and are known as Streptococcus infectious bacteria other than group A. For example, group D. faecalis (type 7 hemolysis), S.faecium (type a hemolysis) is called enterococcus, but a few cystitis and renal pelvis can cause urinary tract infections. Inflammation or endocarditis. Recently, severe enteritis caused by nosocomial infection of VRE that is resistant to vancomycin has become a problem. VRE is resistant to vancomycin developed by a pharmaceutical company in the United States in 1962. Drug-resistant Enterococcus was discovered in France in 1988. Since then, nosocomial infections have been increasing in hospitals or elderly facilities in North America throughout Europe. Even in Japan in July this year, VRE infections caused patients to die. News report. VRE itself is weak in pathogenicity, and there is no question about it in healthy human body. 'However, it will be guided in the body of low-resistance localized or critically ill ____ 4 This paper is suitable for financial standards (CNS) A view frame (21GX297 mm ------------- (Please read the precautions on the back before filling this page)

586931 A? ___________ V. Description of the Invention (2) — One 'caused death. If the patient has VRE bacteria and a large amount of vancomycin is used in the treatment, although other bacteria can be inhibited, VRE may be proliferated. On the other hand, since drugs other than vancomycin are difficult to have an effect on enterococci, there is currently a problem of lack of drugs for enterococcal treatment. [Problem to be Solved by the Invention] In view of the above-mentioned redundant matters, an object of the present invention is to provide an antibacterial agent having an effect on enterococci, especially VRE. [Method for solving the problem] In order to achieve the above object, the focus of the enterococcus antibacterial agent of the present invention is that it contains royal jelly and / or Gandema lucidum fruit body (including umbrella and stem Both). The above perennial mushrooms are mature and contain most spores. The anti-edge agent of the present invention is an antibacterial agent having antibacterial effect on vancomycin-resistant enterococci of enterococci. [Embodiments of the Invention] The present inventors have studied antibacterial agents which can exhibit extremely good antibacterial effects, such as enterococci, especially vancomycin-resistant enterococci (hereinafter, also indicated by VRE). As a result, it was found that the extract of the umbrella part of the royal jelly and the perennial fruit body (also known as ganoderma lucidum) has good antibacterial effects; these alone have a good effect, but the combined use of the two has an 'antibacterial effect. Remarkably improved, and finally completed the present invention. First, the antibacterial effect of royal jelly on enterococci will be explained. The royal jelly that can be used in the present invention is not particularly limited, but it must be confirmed that its antibacterial power will have some differences depending on the place of production, pollen properties, etc. _ 5 This paper is in general Ningguo National Standard (CNS) A4 format (21 × 297 mm) Read First Read Back 1¾ Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs ί Item

V. Description of the invention (3) ^ ~ 'one one… one…-Secondly, the antibacterial effect of Ganoderma on Enterococcus is explained. + The present inventors conducted extensive research on Ganoderma lucidum, and found that Ganoderma lucidum has anti-diarrheal effect of MG, antibacterial effect of streptococcus, and bulimia can enhance trauma treatment effect, and has completed a patent application, which is a Staphylococcus antibacterial 2. Streptococcus antibacterial agents have been approved. Later, as a result of further research using Ganoderma lucidum, it was recently discovered that Ganoderma lucidum has extremely good antibacterial effect on enterococci, especially VRE, and completed the present invention. The extract 0 of the umbrella part of Ganoderma lucidum used in the present invention will be described in detail below. First, the cultivation of Ganoderma lucidum is carried out by the "log cultivation method" using a log of the beech family. Since the selection of the log can play the role of the present invention, it is extremely important. Specifically, it is preferred to cut hardwood ladders, beech, chestnut, and other cypresses of the beech family xylem into short pieces of 10 to 20 cm. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. After sterilizing the short wood, inoculating the bacteria, then growing the bacteria in a stable temperature environment, and then allowing the bacteria to survive for a sufficient time until the fungus circulates throughout the xylem until. The short wood in which the above-mentioned fungi are buried is buried in the soil, and then water is sprayed moderately and repeatedly, and the ganoderma is grown while adjusting the temperature, light amount, and air volume. The present invention uses the above cultivation method to cultivate Ganoderma lucidum. It is very important to use the large-scale sawdust pin cultivation instead of raw wood or the growth and growth of natural objects. The umbrella part will have a strong taste. The best Ganoderma lucidum has been experimentally confirmed to be unable to exert the desired effect. This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210X297 male 586931 Λ7 ΙΓ) 5. Description of the invention (4) (Please read the notes on the back before filling this page) Among the ganoderma lucidum cultivated as described above, the most Those who are mature and contain a large number of spores are used. In other words, in the time and place of the ganoderma used in the present invention, it is best to use "developers in the lower layer (fruit layer) of the umbrella body of the mature fruiting body filled with spores." In other words, the highest period of spore production (the state in which the seed layer is full of spores) is the optimal period for Ganoderma lucidum harvest. On the other hand, after the mature period, the spores will escape from the openings of the seed layer, so the residual spores will change The antibacterial effect is significantly reduced. This is also confirmed experimentally. Therefore, specifically, the umbrella portion of the ganoderma lucidum used in the present invention has a diameter of 14 to 17 cm, a thick meat (thickness of about 35 mm or more), and a dry weight after removing the stem portion. A shape of about 100g • It looks better, and it is better to choose ganoderma filled with spores. It is to extract this ganoderma. The extraction method is, for example, to After slicing the umbrella of Ganoderma lucidum, it is extracted in hot water with a temperature of 95 ~ 100 ° C, and the extract with the highest concentration is filtered, and then the spray drying method is preferred. Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs In order to make the present invention exhibit the most effective antibacterial effect of enterococci, it is best to use a mixture of the above-mentioned extract of Ganoderma lucidum and royal jelly. With the mixture, the antibacterial effect of ganoderma and royal jelly can be multiplied, The antibacterial effect of Enterococcus is very good. The antibacterial agent of the present invention has effects on Enterococcus, that is, bacteria of the genus Enterococcus, such as Enterococcus faecalis (Stool Feces), Enterococcus faecium (Enterococcus), Enterococcus.durans (Intestine Coccus), Enterococcus.avium (Enterococcus 7 This paper size applies to the Chinese National Standard (CNS) A4 specification (21〇 > < 297mm) 5. Description of the invention (5) bacteria), VRE (Vancomycin Resistant Enterococcus), especially for the wild strain E.faecium from human origin VRE also has an antibacterial effect. Therefore, the antibacterial agent of the present invention can be widely applied to Enterococcus belonging to the heat resistance and salt tolerance. The dosage form of the antibacterial agent of the present invention is not particularly limited, but it can be turned into a powder, capsule, or liquid (Generally refreshing water drinks, etc.) for internal use, etc. In addition, in order to further enhance the antibacterial effect on VRE, the umbrella extract of Ganoderma lucidum and the royal jelly were mixed in approximately equal amounts, and then added with oligosaccharides, or accompanied by It is better to add a small amount of charcoal powder for the symptoms of chancre. In addition, when applied to human oral preparations, the dosage of the above-mentioned mixture of dry powder of Ganoderma lucidum and royal jelly is 15 to 20 mg / kg / day, divided into 2 to 3 doses. good. The antibacterial agent according to the present invention may also contain vitamin C, vitamin E, vitamin K, folic acid or excipients, stabilizers, sweeteners, etc. depending on the dosage form. Hereinafter, the present invention will be described in detail using examples. However, the present invention is not limited to the following embodiments, and the embodiments may be modified without departing from the scope described before and afterwards, and all are included in the technical scope of the present invention. [Example] Example 1 [Preparation of Ganoderma Umbrella Extraction Liquid]

The mature Ganoderma lucidum stems full of spores obtained by the above-mentioned log house cultivation method are removed, and only the umbrella portion is cut into thin slices, and then pulverized to obtain a crushed umbrella portion. Add 2L to 100g of this crushed material. The paper size is applicable to Chinese National Standard (CNS) A4 specifications (210x 297); (Please read the precautions on the back before filling this page) 丨 Order, 4 586931 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Λ7 V. Invention Description (6) The water is heated for several hours, and then the highest concentration of the extracted liquid is used as the stock solution, and the stock solution itself and the 1/2 concentration of the stock solution (2 Dilution), 1 / 10th of the concentration of the original solution (10-fold dilution), total 3 types of extracts. [Preparation of royal jelly] Royal jelly (100% raw) is called the original matrix gum. The dilution is performed using sterilized distilled water. Three types of raw matrigel itself, 1/2 concentration (2-fold dilution) of the original matrigel, and 1/10 concentration (10-fold dilution) of the original matrigel were prepared. [Preparation of Ganoderma + Royal Jelly Mixture] Prepare the same amount of mixture of the above-mentioned extract of Ganoderma lucidum (stock solution) and royal jelly (100% raw) (referred to as "stock solution" in Table 1 below); The same amount of mixture of Umbrella extract (2-fold dilution) and royal jelly (2-fold dilution) (referred to as "2-fold dilution" in Table 1 below); the umbrella extract (10-fold dilution) of Ganoderma Lucidum and royal jelly (10-fold dilution) of an equal amount of mixture (abbreviated as "10-fold dilution" in Table 1 below). [Test bacteria]

Enterococcus faecalis

Enterococcus faecium

Vancomycin Resistant Enterococcus (VRE) faecalis Vancomycin Resistant Enterococcus (VRE) faecium antibacterial test is a susceptibility test using the disc method, that is, a Halo test. In detail, each sample prepared in the above manner was immersed in a disc of 8 mm in diameter manufactured by the Showa disc, and this disc was placed on 9 pieces of paper coated with a culture medium for removing bacteria from each colony tested. The scale is applicable to the Chinese National Standard (CNS) Λ4 specification (210X29? Public address) (Please read the precautions on the back before filling out this page),-Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economy 586931 Λ? ----- ----------____ Fifth, three positions on the description of the invention (7), and then cultured at 37 ° C for 24 hours, and then measure the width of the development prevention circle. In addition, the medium used was Enterococcus medium, and three plate culture mediums were prepared for each strain for the test. A total of nine development preventing circles were measured (excluding the measured value of the disk diameter) to calculate the average value. The results paid are shown in Table 1. Developmental block Ganoderma lucidum extract royal jelly ganoderma + royal jelly collagen solution 2 times diluted 10 times diluted original solution 2 times diluted 10 times diluted original solution 2 times diluted 10 times diluted (cm) E.faecalis 19.6 9.3 0.9 19.7 9.4 0.9 22.5 12.0 2.5 E. faecium 19.7 9.5 1.0 19.7 9.5 0.9 23.5 12.7 2.7 VRE (faecalis) 20.0 11.0 0.8 19.6 9.2 0.8 22.5 12.5 2.6 VRE (faecium) 20.0 10.5 0.8 19.7 9.5 0.9 23.7 12.8 2.8 According to Table 1, if we use the extract And / or royal jelly, it has a good antibacterial effect on any type of enterococcus including VRE. Furthermore, when a mixed solution of the two is used, the antibacterial effect on Enterococcus is significantly enhanced. Example 2 This example measures the sterilization status of extracts (original solution) and royal jelly (original solution) of Ganoderma lucidum in the suspension liquid of various enterococci. First, in the same manner as in Example 1, a specific mixture of the extract of Ganoderma lucidum and royal jelly was prepared. On the other hand, it is intended to be used in Example j When _ 10 This paper size applies the Chinese National Standard (CNS) M specification (210X297 ^ 11 ------ 'a ~ (Please read the precautions on the back before filling this page }

Λ7 1— -— ~ 一 ________jy Description of the invention (8 ~ — ............— ~ ------ ~ — ^ 4 kinds of miscellaneous for test _ (about 2mg / ml). They were diluted to 1G times, 2Q times, ⑽ times, 5ig times, iggq times, and 10,000 times. # ， 身 其 夂, add 5mi of the above-mentioned bacteria in a test tube, and then add 1ml The above-mentioned mixture of Ganoderma lucidum and royal jelly is equivalent, then the test tube is sealed with a film, and then placed in a constant temperature bath at 37 ° C. After a predetermined time, it is subjected to sedimentation treatment, and then smeared on a plate medium for cultivation to produce colonies or not. Determine the life and death of the bacteria. The results are shown in Table 2. —--- M—— 丨 (Please read the precautions on the back before filling this page) E.faecalis printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (Hours) VRE (faecalis) Elapsed time (hours) Dilution factor 6 12 24 48 6 12 24 48 10 Life, death, life, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death, death: Everyday 1000 Everyday Everyone 5000 Everyday Foolproof Foolproof 1000 Foolproof-Fake paper standards apply to Chinese National Standard (CNS) A4 specifications (210X29? Public seal) 586931 Λ? ___ ΙΠ ------V. Description of the invention (9) —— the strain li E.fae i is out of date f cium 3 (hours) ----- ---- VRE (faecium) elapsed time (hours) dilution factor 6 12 24 48 6 12 24 48 10 life and death Fate Fate 20 Fate Fate 100 Fate Fate 500 Fate Fate 500 Fate Fate Fate 1000 Fate Fate 5,000 Fate Fate 5,000 Fate Fate 1000 Fate Fate It is printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs that according to Table 2, it can be known that even when a bacterial suspension liquid with a low dilution rate (10 to 20 times) is used, it is treated with the antibacterial agent of the present invention. The treatment for ~ 24 hours can kill enterococci, so it can be confirmed that the antibacterial agent of the present invention has extremely good antibacterial effect. [Effects of the Invention] The present invention is constituted as described above, and can provide the ability to produce enterococci, especially VRE. Very good antibacterial effect of antibacterial agents. ___ 12 paper scale applicable Chinese National Standard (CNS) Λ4 Specification (2ΐ〇χ 297 ^ ΪΤ (Please read the Notes on the back page and then fill in}

Claims (1)

586931 No. 87119409 Patent Application Amendment A8 B8 C8 D8 Revision Date: 92.9.12 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 6. Application for Patent Scope 1. A method for manufacturing enterococcal antibacterial agents, which includes the following steps: The extract from the umbrella body of the fruit body of Royal jelly and / or Ganoderma lucidum is mixed to obtain the enterococcus antibacterial agent; wherein the extraction system of the umbrella body of the fruit body of the perennial mushroom is extracted by the following method: A process of slicing the umbrella body of the perennial mushroom body; a process of grinding the slice of the fruit body umbrella portion; a process of extracting the ground and pulverized fruit body umbrella portion for several hours in hot water ; The extraction liquid is subjected to a spray drying process. 2. The method for producing an enterococcus antibacterial agent according to item 1 of the scope of the patent application, wherein the above-mentioned ten thousand years are mature and contain a large number of spores. 3. The method for producing an enterococcus antibacterial agent according to item i or 2 of the scope of application for a patent, wherein the enterococcal antibacterial agent has an antibacterial effect on vancomycin-resistant enterococci. 4. An enterococcus antibacterial agent, which is mainly composed of an extract from the umbrella body of the fruit body of Royal jelly (Royal jelly) and / or Ganoderma lucidum. The aforementioned royal jelly extract includes protein, carbohydrate, Fat, minerals, and water; The extracts of the fruit body of the perennial mushroom include ganoderma acid A and ganoderic acid C1 + H, and the extracts of the fruit body of the perennial mushroom are extracted by the following methods: (Please fill in the precautions on the back of Mtf first! Ϋα • Line 586931 A8 B8 C8 D8 The scope of patent application for the process of slicing the umbrella body of the perennial mushroom body; Grinding the slice of the umbrella body of the fruit body Manufacturing process: the process of extracting the ground portion of the ground fruit body and then extracting it in hot water for several hours; manufacturing process of extracting the extract solution by spray drying. 5 · the intestine as described in item 4 of the scope of patent application Coccus antibacterial agents, in which the above-mentioned perennial mushrooms are mature and contain a large number of spores. 6 · The enterococcal antibacterial agent described in item 4 or 5 of the scope of the patent application, which has an antibacterial effect on vancomycin-resistant enterococci. 4 η Remarks on the back of the reprint line Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 2 This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)