KR20100007122A

KR20100007122A - Anti-bacterial or anti-fungal composition

Info

Publication number: KR20100007122A
Application number: KR1020080067609A
Authority: KR
Inventors: 한만덕; 백종윤; 김용현; 권현정; 맹주양
Original assignee: 순천향대학교 산학협력단
Priority date: 2008-07-11
Filing date: 2008-07-11
Publication date: 2010-01-22

Abstract

PURPOSE: An antifungal or antibacterial composition containing Caesalpinia sappan L. extract is provided to have high sensitivity to bacteria and fungi and relieve diseases caused by bacteria or fungi. CONSTITUTION: An antibacterial or antifungal composition contains Caesalpinia sappan L. extract as an active ingredient. The composition has antibacterial activity to gram-positive bacteria including Streptococcus faecium, Staphylococcus aureus, and Propionibacterium acnes. The composition also has antibacterial activity to gram negative bacteria including Klebsiella oxytoca, Klebsiella aerogenes, and Enterobacter cloacae. The composition has antifungal activity to Candida albicans.

Description

Anti-bacterial or Anti-fungal Composition

The present invention relates to an anti-bacterial or anti-fungal composition exhibiting excellent antibacterial or antifungal effects against various bacteria or fungi.

Natural antibiotics are selected from natural or microbial sources to maximize biochemical and physiological differences, thereby selectively controlling viruses, bacteria, yeasts, molds, etc., which are harmless to humans or minimize toxicity. do. Among natural antibiotics, especially plant-derived antimicrobials have been used as a source of development of new therapeutic drugs. This is because plants produce antimicrobials in the most efficient way as a defense mechanism for keeping them safe from infectious diseases in the wild.

Joiner ( Caesalpinia sappan L. ) is a deciduous shrub legume distributed in tropical Asia, including India, Malaysia, and southern China. The wood is hard and the heart is bright red. Used for medicinal purposes, bark and fruit have pigments, which are used as red dyes. The dye component is contained in the core, and the main component of the pigment is brazilein (6a, 7-dihydro-3,6a, 10-trihydroxy-benz [b] indeno [1,2-) belonging to the benzopyrane family. d] pyran-9 (6H) -one).

The main components of the joiner are brazilein and sappanchalcone (Formula 1). Especially, brazilian is a compound that accounts for 2% of the total joiner, and can be oxidized in air to form brazilin (Formula 2). It turned out.

Formula 1

Formula 2

The present invention aims to solve the technical problem that has been requested from the past. Specifically, it is an object of the present invention to provide an antimicrobial or antifungal composition which is highly sensitive to various bacteria and fungi and can exert an excellent antimicrobial or antifungal effect.

The antimicrobial or antifungal composition according to the present invention for achieving this object is composed of including the joiner extract as an active ingredient.

The antimicrobial or antifungal composition according to the present invention is highly sensitive to various bacteria and fungi, and can exert an excellent antimicrobial or antifungal effect, thereby exhibiting an improvement effect of a disease caused by bacteria or fungi.

The inventors of the present application, in the case of an antimicrobial or antifungal composition comprising a seedling extract as an active ingredient, the seedling extract active ingredient shows a relatively low minimum inhibitory concentration (MIC) of various bacteria and fungi, It was found that it can exhibit antibacterial and antifungal effects, and it was confirmed that there is an excellent improvement effect of diseases caused by bacteria and fungi.

The composition exhibits antimicrobial activity, for example, may be active against Gram-positive bacteria or Gram-negative bacteria, and as shown in the following examples, the composition according to the present invention is particularly effective against Gram-positive bacteria. It can be seen that it works.

The Gram-positive bacteria is, for example, Streptococcus faecium Staphylococcus aureus ( Staphylococcus) aureus ), Propionibacterium acnes : KCTC 3314) and one or more selected from the group consisting of Streptococcus mutans (ATCC 25175), preferably, as demonstrated in the following examples, Streptococcus faecium MD8b ( Streptococcus faecium MD8b), Staphylococcus aureus Reus Staphylococcus aureus SG511, Staphylococcus aureus 503 ( Staphylococcus aureus 503), Propionibacterium acnes : KCTC 3314) and caries ( Streptococcus mutans : ATCC 25175) can exhibit excellent antimicrobial activity.

In one preferred embodiment, the composition is Streptococcus faecium MD8b, Staphylococcus aureus Staphylococcus aureus SG511 and Staphylococcus aureus When acting on 503 ( Staphylococcus aureus 503), the minimum inhibitory concentration of the composition for exerting antimicrobial activity against each Gram-positive bacterium may be 0.01 to 200 mg / ml. In addition, when the composition acts on P. acnes to give antibacterial activity, the minimum inhibitory concentration (MIC) of the composition against P. acnes may be 40 to 200 mg / ml, When the composition acts on S. mutans and exhibits antimicrobial activity, the minimum inhibitory concentration of the composition against S. mutans may be 2 to 200 mg / ml.

The composition is a Gram-negative bacterium Escherichia coli coli ), Pseudomonas aeruginosa), Salmonella typhimurium (Salmonella typhimurium), keulrep when Ella oxy cytokine (Klebsiella oxytoca), keulrep when Ella Oh Eroge Ness (Klebsiella aerogenes) and Enterobacter keulroeyi time (be at least one selected from the group consisting of Enterobacter cloacae), and , Preferably Escherichia coli DC 2, Pseudomonas aeruginosa 1592E, and Klebsiella oxytoca 1082E ( Klebsiella oxytoca 1082E) has an action of inhibiting growth at a concentration of 0.8 mg / ml to 6.3 mg / ml.

Antifungal compositions according to the invention also, Candida albicans (Candida albicans) to act on the fungi can exhibit the activity, the minimum inhibitory concentration for Candida albicans (Candida albicans) of the composition may be from 90 to 200 mg / ml.

The antimicrobial or antifungal composition according to the present invention may be a pharmaceutical composition, a skin cleansing composition, an antibacterial building material and household composition or a cosmetic composition.

The pharmaceutical composition may be administered orally in the form of solid, semi-solid or liquid by adding a commercially available inorganic or organic carrier, or be administered parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, or the like. Can be.

The preparation for oral administration includes tablets, pills, granules, soft and hard capsules, powders, fine granules, powders, emulsions, syrups, pellets and beverages. Etc. can be mentioned. In addition, preparations for parenteral administration include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like.

In order to formulate the active ingredient of the present invention, it can be easily formulated according to a conventional method, and surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffers, suspensions, and other commonly used auxiliaries can be suitably used.

In addition, the dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of one of ordinary skill in the art and generally dosages may range from 0.001 mg / kg / day to approximately 2000 mg / kg / day, although any method is intended to limit the scope of the invention. no.

Specifically, the skin cleansing composition may be a functional cosmetic soap, and the functional cosmetic soap is particularly effective in treating acne of the skin, as it has an antibacterial activity effect against acne bacteria.

The content of the joiner extract included in the cosmetic soap is not particularly limited as long as it contains an antimicrobial or antifungal activity, but may be preferably 0.5 to 2 parts by weight (W / W%) based on the total weight of the soap. Through this, it is possible to solve problems such as discoloration, odor and the like appearing in the soap containing plant components.

The cosmetic composition may be a flexible cosmetics, nourishing cosmetics, massage creams, nutrition creams, packs, gel or skin adhesive cosmetics, and may also be a transdermal dosage form such as lotions, ointments, gels, creams, patches or sprays.

The invention also relates to (a) joinery ( Caesalpinia ) adding water or an organic solvent to sappan L. ) to obtain an extract; (b) filtering the extract to obtain a filtrate; (C) a method for producing the antimicrobial or antifungal composition comprising the step of concentrating and drying the filtrate under reduced pressure.

When water is added to the joiner in the step (a) to obtain an extract, the temperature of the water may be 70 to 120 ℃, preferably 80 ℃ to 110 ℃, it can be extracted by heating in a bath in the temperature.

In one preferred example, the extract has a water temperature of 90 ° C. to 110 ° C. and is obtained under high pressure conditions of 0.3 kg / cm ² to 0.5 kg / cm ² , preferably 0.318 kg / cm ² to 0.429 kg / cm ² . Can be. As such, when the temperature is controlled to a high temperature and the extraction is performed by adding a high pressure condition, the yield is remarkably increased as compared with the method of extracting the joiner by heating the bath at a normal heating, as evidenced by the following experimental example. You can check it.

When the organic solvent is added to the joiner in the step (a) to obtain an extract, the organic solvent may be at least one selected from the group consisting of hexane, chloroform, ethyl acetate, butanol and methanol.

At this time, the extraction is, for example, hexane and water are added to the extract extracting the joiner with methanol, the hexane layer and the water layer is separated, and then the water layer can be further separated, to obtain a hexane extract of the joiner. By adding chloroform to the layer separated from the hexane layer, the chloroform extract is obtained, and ethyl acetate is added to the upper layer separated by the chloroform layer, followed by extraction to obtain the ethyl acetate extract. Butanol extract may be obtained by adding butanol to the layer separated by ethyl acetate, and finally, the water layer separated by butanol may be concentrated to obtain a water fraction. In addition, methanol extract can be obtained by separating the first extract extracted with methanol.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the following Examples are provided to illustrate the present invention, and the scope of the present invention is not limited thereto.

Example 1 Preparation of Joiner Extract-Hot Water Extraction

First, the seedlings were hydrothermally extracted. In the hot water extraction method, the core material was pulverized and quantified by 50 g, then placed in a Erlenmeyer flask, 1 L of distilled water was added to submerge the sample, and then extracted at a water bath using a reflux cooler at 100 ° C. for 1 hour. The filtrate was obtained by filtration, and the residue was subjected to extraction and filtration again to obtain a filtrate. The filtrates were combined, dried under reduced pressure using a rotary concentrator, and water extract was prepared.

In order to obtain a higher yield, the extraction was carried out using an autoclave under high pressure for 1 hour at 100 ℃, the results are shown in Table 1. In this way, extraction with a 100 ° C. autoclave yielded an increase in yield of 68.3% over 80 ° C. extraction.

[Table 1] Yield variation according to the method of extracting hot water of joiner

(Yield (%) = dry weight of extract (g) / dry weight of joiner (g) X 100)

Example 2 Preparation of Joiner Extract-Organic Solvent Extraction

Joiner extraction was carried out through an organic solvent, the extraction process is shown in FIG. First, 1 v / v was added to the separating funnel in a separating funnel, and the ratio of 10 v / v of hexane and 9 v / v of water (methanol extract: water: hexane = 1: 9: 10) was added. Each solvent was shaken repeatedly to mix well, and then allowed to stand to separate into hexane and water layers. The water layer formed on the lower layer was well separated and stored in another separatory funnel, and the organic solvent hexane was added and extracted three times.

The separated hexane layer was concentrated under reduced pressure at 80 ° C. to obtain a hexane extract of joiner (SIP-1). The lower layer separated from the hexane layer was extracted by adding chloroform 1: 1 v / v, and extracted three times, thereby obtaining a chloroform extract (SIP-2) by concentrating under reduced pressure at 80 ° C. The ethyl acetate was added 1: 1 v / v to the upper layer separated by the chloroform layer, mixed well, extracted three times, and concentrated under reduced pressure at 80 ° C. to obtain an ethyl acetate extract (SIP-3). The lower layer separated with ethyl acetate was added with a 1: 1 amount of butanol, mixed, and extracted three times. The separated upper layer was concentrated under reduced pressure at 80 ° C. as a butanol layer to obtain a butanol extract (SIP-4). The water layer finally separated with butanol was concentrated under reduced pressure at 80 ° C. to obtain a water fraction (SIP-5). In addition, the seedling extract first extracted with methanol was separated at 80 ℃ to obtain a methanol extract (SIP-6) was used in the activity test, the yield of each extract is shown in Table 2 below.

[Table 2] Yield of joiner extract

Referring to Table 2, when fractional extraction of the dried seedlings from various organic solvents, methanol, water, butanol, ethyl acetate, chloroform, hexane extracts showed high yields in that order. The purity and absorbance of the ethyl acetate extract used in the experimental example is shown in Figures 2 and 3, the purity of the ethyl acetate extract was 96%, the main absorbance was confirmed to be 215nm, 257.6nm, 287.2nm.

Experimental Example 1 Screening Test of Antimicrobial Activity by Joiner Extract

The strain used for the antimicrobial experiment was S. mutans ATCC 25175, and Fleisch extract broth (Difco, USA) was used for all strains except S. mutans ATCC 25175 as a growth medium. Incubate for 18-24 hours. S. mutans ATCC 25175 bacteria were cultured in BHI (Brain heart infusion, Difco, USA) medium after 1st to 2nd subculture, and then incubated in the same medium at 37 ° C for 48 hours under anaerobic conditions. MHA (M ㆌ eller hinton agar, Difco, USA) was used for the antimicrobial activity experiment.

The antimicrobial activity screening experiment of the extract of the joiner used a paper disc method. The S. mutans ATCC 25175 strain used for antimicrobial screening was inoculated into 1 ml of each strain cultured on a slope medium and inoculated into bacterial growth medium in 10 ml BHI (Brain heart infusion, Difco, USA) medium to activate for 15-18 hours. Antimicrobial test plate medium was hardened by dispensing 15 ml each into a culture dish using sterile MHA medium. 50 μl of each strain previously activated on the dried MHA medium was plated with a cotton swab. A sterilized filter paper disc (filter paper disc-8 mm: 1.5 mm, Toyo Roshi Kaisha, ltd., Japan) was placed on the surface of the inoculated medium and adhered to it. 100 μl each was absorbed. The medium in which the seedling extract was absorbed into the disk was left to stand at room temperature for 15 minutes to dry the disk, and then incubated at 37 ° C. under anaerobic conditions for 15 to 18 hours. After incubation, the size of the inhibitory zone diameter (mm) produced around the disc was measured by a vernier caliper (Mitutoyo, Japan), and the degree of antimicrobial activity of each extract was shown in FIG. 4.

Referring to Figure 4, joiner extract (SCH-29) is Compared to the genus extract (SCH-30: Phlomis umberosa ) , Equisetum hiemale (SCH-31: Equisetum hiemale ) , and equestrian extract (SCH-32: Cimicifuga heracleifolia ), it showed the lowest activity of 36mm against S. mutans ATCC 25175. You can see that it shows.

Experimental Example 2 Secondary Antimicrobial Activity Test by Joiner Extract

The minimum inhibitory concentration (MIC) was determined by a continuous dilution method in agar medium, and the antimicrobial activity against S. mutans ATCC 25175 in the agar medium was determined by the paper disk method. As in 3, MICs (minimum inhibitory concentrations) for 18 pathogenic strains including S. mutans ATCC 25175 (4 Gram-positive bacteria and 14 Gram-negative bacteria) were measured.

BHI medium (Brain heart infusion broth) and Fleich extract broth were used for S. mutans ATCC 25175 and 18 pathogenic strains, respectively. All used MHA medium. Each strain 1 loop (l loop) was inoculated in a 10ml liquid medium, and then used by incubating for 24 to 48 hours at 37 ℃. Each joiner extract was diluted to the 17th consecutively with an initial concentration of 10 mg / ml until the next concentration was 1/2 of the initial concentration, and then mixed and hardened together before dispensing on the plate medium. The strains were inoculated in a constant concentration plate and incubated for 24 hours at 37 ° C. for anaerobic or aerobic incubation, and then visually determined to determine the proliferated and inhibited concentrations, and the results are shown in Table 3. It was.

Table 3 Minimum Inhibitory Concentration Test of Joiner Extracts (MIC test)

S. mutans ATCC 25175 and 18 kinds of pathogenic results of the measurement of the relative minimum inhibitory concentrations of the joiner extract having a high antibacterial activity against the strain, showed a higher effect in Gram-positive bacteria, S. feacium MD8b, SG511 S. aureus and S. The bacteria of aureus 503 showed the minimal inhibitory effect at the concentration of 0.2 mg / ml. In Gram-negative bacteria, E. coli DC 2, P. aeruginosa 1592E and K. oxytoca 1082E showed minimal inhibitory effects at concentrations of 0.8 mg / ml. As a result of measuring the minimum inhibitory concentration, the extract of the joiner was confirmed to exhibit particularly excellent activity in Gram-positive bacteria.

Experimental Example 3 Antiacne and Antifungal Activity Test

The antifungal effect of the fungus ethyl acetate extract on the fungus Candida albicans KCTC 7965 is shown in Tables 4 and 5 below, and FIGS. 5 to 7. Referring to Table 4 and Figure 5, it can be seen that the antifungal effect is shown in the medium of 200 mg / ml concentration. Referring to Table 4 and Figure 6, the antibacterial effect against the acne bacterium P. acnes KCTC 3314 was shown at 50 mg / ml. In addition, referring to Table 4 and Figure 7 in the test for the caries S. mutans ATCC 25175 showed a growth zone of 41 mm at a concentration of 25 mg / ml.

[Table 4] Antimicrobial Effects of Acanthus, Fungi and Caries on the Joiner Extracts

* ND: no antimicrobial activity

[Table 5] Minimum Inhibitory Concentration of Joiner (MIC)

As shown in Table 5, the Candida KCTC 7965 strain had an antifungal effect at 100 mg / ml concentration, the acne bacterium KCTC 3314 strain at 50 mg / ml concentration and the caries ATCC 25175 strain at 3.1 mg / ml concentration. It can be confirmed that the suppression.

Therefore, it can be seen that the composition according to the present invention has excellent antibacterial and antifungal efficacy against various bacteria and fungi.

Example 3 Preparation of Soap

Soap was prepared in the composition shown in Table 6 so as to maintain the antibacterial effect by diluting the selected ethyl acetate extract of the seedlings by concentration.

Table 6 Composition of soap

Experimental Example 4 Test Evaluation of Soap

1. Evaluation of the feeling of soap

Example 1 (containing the joiner extract) and control (soap without the joiner extract) were tested as follows. That is, using the joiner soap (Example 1) and the soap prepared by Comparative Example 1, the acne improvement effect, the feeling of use and the like were evaluated for those who have acne on the skin. The comparison test and the acne improvement effect comparison test was conducted on the soap of soap and soap of Comparative Example 1 in male and female college students with 90 acne.

The method of use was the same as when using normal makeup soap for 30 days, to wash twice a day, and after 30 days to evaluate the results by a self-evaluation method. For the comparison of the feeling of use on the skin, the average score of 90 people was calculated. The evaluation score is 5 points for "skin is very good" than normal makeup soap, 4 points for "skin is better" than general makeup soap, 3 points for "same" with normal makeup soap, and normal makeup soap 2 points for "Slightly rough skin" and 1 point for skin rougher than normal makeup soap "were added, and the percentage of respondents for each item was calculated. For statistical analysis, all data obtained through the survey were used for SPSS WIN statistics program.

[Table 7] Feeling test of soap

As shown in the test evaluation results of Table 7, among the people who used the soap of Comparative Example 1, the respondents who responded positively to "Improvement of skin" and "Improvement" were 44% and 24%, respectively. In the case of soap, 60% responded, indicating that it showed a better feeling than ordinary soap.

Comparative Example 1 is a soap containing all the vegetable extract components except the joiner extract, unlike Example 1 including the joiner extract, there were more positive answers than ordinary soap. This is because general soap stimulates inflammation of acne patients because of its alkaline property, whereas soap according to Example 1 of the present invention has an effect of relieving irritation to the skin by the antibacterial and antifungal effects of joiner extract. .

2. Test of acne improvement effect of soap

In the comparison test of acne improvement effect, 5 points of "Improvement of acne was very good". 4 points of "Improvement of acne." 1 point of "Acne is very deteriorated." 1 point was added for each point, 3 points for no change, 4 points for "the improvement effect of acne", and 5 points for "the improvement effect of acne is very good", and each item was statistically processed. For statistical analysis, all data obtained through the survey were used for SPSS WIN statistics program.

[Table 8] Acne improvement effect test

As can be seen in Table 8, among those who used the soap of Comparative Example 1, 42.3% and 25.0% responded positively to "the improvement of acne" and "the improvement of the acne was very good", respectively. On the other hand, in the case of using the soap of Example 1, in response to 67.3%, it can be seen that there is an effect of relieving acne than the soap of Comparative Example 1. In particular, Example 1 containing the joiner extract shows an improvement of 36.3% over the soap of Comparative Example 1 does not include the joiner extract.

Example 4 Preparation of Lotion

Joiner extract of Example 1 3.00

L-ascorbic acid-2-magnesium phosphate salt 1.00

Water Soluble Collagen (1% Aqueous Solution) 1.00

Sodium Citrate 0.10

Citric Acid 0.05

Licorice Extract 0.20

1,3-butylene glycol 3.00

Purified water level

(Unit: weight%)

Example 5 Preparation of Cream

Joiner Extract 1.00 of Example 1

Polyethylene Glycol Monostearate 2.00

Self-emulsifying glycerin monostearate 5.00

Cetyl Alcohol 4.00

Squalene 6.00

Tri2-ethylhexaneglyceryl 6.00

Sphingolipids 1.00

1.3-butylene glycol 7.00

Purified water level

(Unit: weight%)

Example 6 Preparation of Pack

Joiner extract of Example 1 5.00

Polyvinyl Alcohol 13.00

L-ascorbic acid-2-magnesium phosphate salt 1.00

Lauroylhydroxyproline 1.00

Water Soluble Collagen (1% Aqueous Solution) 2.00

1,3-butylene glycol 3.00

Ethanol 5.00

Purified water level

(Unit: weight%)

Those skilled in the art to which the present invention pertains will be able to perform various applications and modifications within the scope of the present invention based on the above contents.

1 is a schematic diagram showing a process for preparing an extract from joiner through an organic solvent;

2 and 3 are graphs showing the results of measuring the purity and absorbance of the joiner extract extracted with ethyl acetate;

4 is a graph showing the results of the antimicrobial activity of each extract by measuring the size of the inhibition zone diameter (mm) with a vernier caliper (Mitutoyo, Japan);

5 to 7 are photographs showing the anti acne and antifungal activity test results of the fungi, acne and cavities of the composition according to an embodiment of the present invention.

Claims

Anti-bacterial or anti-fungal composition comprising a joiner extract as an active ingredient.

The method of claim 1, wherein the composition exhibits antimicrobial activity against one or more Gram-positive bacteria selected from the group consisting of Streptococcus faecium , Staphylococcus aureus , and Propionibacterium acnes . Antimicrobial or antifungal composition, characterized in that.

According to claim 1, wherein the composition is Escherichia coli, Pseudomonas aeruginosa , Salmonella typhimurium, Klebsiella oxytoca, Klebsiella aerogenes (Klebsiella aerogenes) And Enterobacter cloacae ( Enterobacter cloacae ) Antimicrobial or antifungal composition, characterized in that it exhibits antimicrobial activity to one or more Gram-negative bacteria selected from the group consisting of.

According to claim 1, wherein the composition is antimicrobial or antifungal composition, characterized in that the activity by acting on acne ( P. acnes ).

The antimicrobial or antifungal composition of claim 1, wherein the composition exhibits activity by acting on Candida albicans fungi.

(a) adding water to Caesalpinia sappan L. at a temperature of 85 to 120 ° C. under high pressure of 0.3 kg / cm ² to 0.5 kg / cm ² , or adding an organic solvent to obtain an extract;

(b) filtering the extract to obtain a filtrate;

(C) a method for producing a composition for anti-bacterial or anti-fungal according to claim 1 comprising the step of concentrating and drying the filtrate under reduced pressure.

The method of claim 6, wherein the organic solvent is at least one selected from the group consisting of hexane, chloroform, ethyl acetate, butanol and methanol.