KR20190047182A - Composition for sterilization or disinfection comprising carnosic acid, carnosol or glabridin as active ingredient - Google Patents

Abstract

The present invention relates to a composition for disinfection containing carnosic acid, carnosol, or glabridin as active components and, more specifically, to a composition for disinfection containing any one compound or two or more compounds selected from a group consisting of carnosic acid, carnosol, glabridin, and salt thereof as active components. The composition for disinfection by the present invention can be safely used for the disinfection of food, groceries, devices for food, or animal skin since mankind uses natural materials separated and purified from plants whose safety is proven by being used as cosmetics or medicine and planted by mankind for long time. The composition of the present invention can be used as a disinfectant by having excellent disinfection properties with a process of short time at a lower concentration to be distinguishable from previously known antibiotics. Moreover, the present invention has the remarkably low risk of developing bacteria resistant to existing antibiotics by killing microorganisms in a short time.

Description

TECHNICAL FIELD The present invention relates to a composition for sterilizing and disinfecting carnosic acid or carnosol or glabridin as active ingredient containing carnoic acid,

The present invention relates to a composition for sterilizing and disinfecting a carnosic acid, a carnosol, a glabridin and a combination thereof, which comprises a carnosic acid, a carnosol or a glabridin as an active ingredient. And salts thereof as an active ingredient. The present invention also relates to a composition for sterilization and disinfection containing one or more compounds selected from the group consisting of salts thereof.

Disease patterns and incidents of foodborne illness and accidents are increasing as the income level of the economy grows and the dietary pattern is diversified and the consumption of food and instant food increases. According to the WHO report, more than 33 million people worldwide are exposed to food poisoning annually due to rising temperatures due to global warming, and are now becoming a public health risk factor for humans. Also, due to the recent humidifier disinfectant, hamburger disease, insecticide egg, and sanitary noxious substance controversy such as humidifier disinfectant in Korea, there is a growing interest in the sense of rejection and public hygiene in synthetic preservatives and synthetic disinfectants.

Currently, various disinfecting products for domestic and foreign purposes are sold and sold. However, since they do not meet the sanitization and disinfection test standards stipulated in the Ministry of Food and Drug Administration and contain harmful substances in the human body, the safety problem of sterilization disinfectant still remains as a social issue It does not meet consumer demand for environmentally friendly non-toxic disinfectant.

The disinfection test method disclosed in the Food Additives Code is a test method different from the method for measuring the MIC of the antimicrobial agent specified in The Clinical & Laboratory Standards Institute (CLSI). The sterilization test is a method for measuring the reduction rate (%) of viable cells (cfu / ml) against the initial number of bacteria (cfu / ml) by reacting the test substance with the target microorganism for 5 minutes and measuring the MIC value of the antibacterial agent specified in CLSI Is a method of measuring the minimum concentration that inhibits microbial growth after a reaction of 16 hours or more by mixing a test substance with a target microorganism. Both test methods are the same in terms of measuring the ability of the test substance to inhibit microbial growth. However, the reaction time between the test substance and the target microorganism is very different from that of the sterilization disinfection test method of 5 minutes and the MIC measurement method is more than 16 hours .

The disinfection power is a measure of the unique time-kill rate capability of the antimicrobial material, which is due to the microbial death mechanism that is distinct from the conventional antimicrobial material. For these reasons, antimicrobial substances reported to have antimicrobial activity often have no sanitizing power.

Disinfecting agents such as utensils in food additives should contain an active ingredient which has a disinfecting action against harmful microorganisms. The components are hydrogen peroxide, peroxyacetic acid, citric acid, ethanol, iodine, chlorine dioxide, hypochlorous acid Water, etc., and these components contain danger and harmfulness such as carcinogenicity, corrosiveness, skin irritation and the like.

Drugs for skin treatment by microbial infections are divided into general drugs and specialty drugs. The main ingredients of generic drugs are fuidic acid, mucilage, gentamycin, bacitracin and neomycin. Clinical drugs include clindamycin, erythromycin . These antibiotics are excellent in antimicrobial activity, but they also kill microorganisms by inactivating microbial replication, nucleic acid and protein synthesis, and other major metabolic processes, so that the time-kill rate is more than 24 hours, Can not be killed. In addition, it causes various side effects such as systemic toxicity and generation of antibiotic resistant bacteria, burning, rash, itching, and is not suitable for disinfection composition.

Accordingly, the present inventor has developed a sterilizing and disinfecting composition which can be used as an antiseptic disinfectant by showing excellent disinfecting power even in a short time, and using a natural substance which is not a chemical synthetic substance.

Produce handling and processing practices. Emerg. Infect. Dis. 3: 459-463,1997. Liao CH, Sapers GM. Attachment and growth of salmonella chester on apple fruits and in vivo response of bacteria to sanitizer treatments. J. Food Protect. 63: 876-883,2000. Romanova N, Favrin S, Griffith MW. Sensitivity of Listeria monocytogenes to sanitizers used in the meat processing industry. Appl. Environ Microbiol. 68: 6405-6409, 2002. Jang JH, Jang JS, Lee SY, Kim HS, Kang SM, Park JH. Growth inhibition effects of ethanol and sodium chloride on Bacillus cereus. Korean J. Food Sci. Technol. 35: 998-1002, 2003. Luppers SBI, Reji MW, Van der Heijden RWL, Rombouts FM, Abee T. Development of a standard test to assess resistance to Staphylococcus aureus biofilm cells to disinfectants. Appl. Environ Microbiol. 68: 4194-4200, 2002.

Accordingly, a main object of the present invention is to provide a composition for sterilizing and disinfecting safely using a natural substance which can be used as an antiseptic disinfectant by showing an excellent sterilizing disinfecting power even in a short time of treatment and not a chemical synthetic substance.

According to one aspect of the present invention, there is provided a pharmaceutical composition containing one or two or more compounds selected from the group consisting of carnosic acid, carnosol, glabridin and salts thereof as an active ingredient The present invention provides a composition for sterilization and disinfection.

The composition for sterilization and disinfection of the present invention preferably contains carnosic acid or a salt thereof.

It is preferable that the composition for sterilization and disinfection of the present invention is for disinfecting food, foodstuff, food appliance or animal skin.

According to another aspect of the present invention, there is provided a food prepared by adding the composition for sterilization.

According to still another aspect of the present invention, there is provided a food additive prepared by adding the composition for sterilization.

According to still another aspect of the present invention, there is provided an external preparation for skin prepared by adding the sterilizing composition for sterilization.

According to still another aspect of the present invention, there is provided a cosmetic product prepared by adding the composition for sterilization.

Since the composition for sterilization and disinfection of the present invention uses a natural substance which is separated and purified from a plant in which human beings have been eating for a long time and used as a cosmetic or pharmaceutical material and its safety has been proved, it is possible to sterilize foods, foodstuffs, It can be used safely for disinfection. In particular, the composition of the present invention can be used as an antiseptic disinfectant because it exhibits excellent disinfecting power even at a low concentration and a short time of treatment, in contrast to known antibiotics. It also has the advantage that the risk of developing resistant microorganisms, which are problematic in existing antibiotics, is remarkably low because microorganisms are killed in a short time.

Figure 1 shows the chemical structure of carnosic acid.
Figure 2 shows the chemical structure of carnosol.
Figure 3 shows the chemical structure of glabridin.

The composition for sterilization and disinfection of the present invention is characterized by containing one or two or more compounds selected from the group consisting of carnosic acid, carnosol, glabridin and salts thereof as an active ingredient .

In the present invention, a disinfecting power of the compounds is first confirmed, and a composition for disinfection using them is provided.

The term " disinfection " in the present invention means inhibiting the growth or proliferation of microorganisms within 5 minutes or killing microorganisms, which is distinguished from the usual antimicrobial activity.

The carnoic acid of the present inventor is represented by the following formula (1).

[Chemical Formula 1]

The carnosol of the present invention is represented by the following formula (2).

(2)

The glabridin of the present invention is represented by the following formula (3).

(3)

The carnosic acid (CAS # 3650-09-7) and carnosol (CAS # 5957-80-2) of the present invention are diterpene isolated from Rosmarinus officinalis or Salvia officinalis ) Antioxidant activity and antimicrobial activity as a natural substance of the family, and carnosic acid and carnosol have been reported to inhibit cancer cell growth by inducing apoptosis of cancer cells. In addition, carinoic acid has been reported to have various physiological activities such as anti-viral activity of inhibiting protease activity of HIV (human immunodeficiency virus). However, the disinfecting power of carnosol and carnosol is not yet known.

The glabridin (CAS # 59870-68-7) of the present invention is an isoflavan-based natural substance isolated from licorice ( Glycyrrhiza uralensis Fischer, Glycyrrhiza glabra L.) A variety of physiological activities such as whitening, antibacterial, antiinflammatory, neuroprotection, and atherosclerosis have been reported, but there has been no report on the activity of glaubridin disinfection.

In the present invention, the salt of the compound is preferably in the form of a pharmaceutical, food or cosmetically acceptable salt. It may be used in the form of acid addition salts or salts such as metal complexes such as zinc and iron.

The composition of the present invention preferably contains 200 ppm or more of the carnoic acid, 500 ppm or more of the carnosol, and 500 ppm or more of the glabridin.

The composition of the present invention may further include other materials as long as it does not significantly lower the disinfecting power of the composition of the present invention. The material to be added at this time is preferably a material which can be used for food or food additives.

The composition of the present invention may contain a solvent for dissolving or dispersing each of the above materials, and may be formulated into various forms such as pharmaceutical, food or cosmetically acceptable.

The composition of the present invention may be added to provide a microorganism as a sterilized food or a food additive capable of disinfecting a microorganism present in the food. It can also be added to external preparations for skin for the purpose of wound healing and the like, and can also be added to cosmetics. At this time, it is preferable that the addition amount of the present invention is 200 ppm or more for carnoic acid, 500 ppm or more for carnosol, and 500 ppm or more for glabridin based on the form applied to the final disinfection object.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example 1. Preparation of composition for sterilization

Carnolic acid (S / N C0609), carnosol (S / N C9617) and glabridin (S / N G9548), which are raw materials for the composition of the present invention, were purchased from Sigma.

Each raw material was dissolved in 100% ethanol and made into a 1% (w / v) stock solution and stored frozen at -20 ° C until the test. Each storage solution was diluted with water or alcohol to a final concentration of 100 ppm, 200 ppm, and 500 ppm of the active ingredient immediately before the sterilization test to prepare a composition in the form of 20% (v / v) ethanol. This was used as a test solution in the following experimental example.

Comparative Example 1. Preparation of Control Composition

As a control for comparison with the composition of the present invention, gentamicin sulfate (S / N G1914), fusidic acid (S / N F0881), bacitracin (S / N 11702 ), Ampicillin trihydrate (S / N A6140), and vancomycin hydrochloride (S / N V1130) were purchased from Sigma-Aldrich.

Each control stock was dissolved in sterile distilled water to make a 1% (w / v) stock solution and stored frozen at -20 ° C until the test. Each of the storage solutions was diluted with water or alcohol to the same concentration as in Example 1 immediately before the sterilization test to prepare a composition in the form of 20% (v / v) ethanol. This was used as a test solution in the following experimental example.

[Test method for sanitizing disinfection]

The disinfection test of the following experimental examples was carried out according to the method specified in No. 2016-32 of the Food and Drug Administration Notice.

The sterilization test was evaluated by the number of viable cells (cfu / mL) reduction rate (%) against the initial number of cfu / mL. The test strain suspension containing the interfering substance is added to the test solution and allowed to react at room temperature for 5 minutes. The appropriate neutralizing agent is then immediately used to inhibit the reaction, and the number of viable cells in each sample is measured in advance to measure the reduction rate of viable cells.

The test solution should be diluted in an appropriate amount by taking a certain amount of the sample.

E. coli ATCC 10536 and S. aureus ATCC 6538 are used as test strains. The test strain is plated on TSA (Tryptic soy agar) medium and cultured at 36 ° C for 18-24 hours. Prepare 10 mL of TSB (Tryptic soy broth) culture in a 100 mL Erlenmeyer flask and transfer the active culture medium of TSA medium using platinum beads. After culturing at 36 ° C for 18 to 24 hours, dilute the solution to adjust the number of live cells to 1.5 to 5.0 × 10 ⁸ cfu / mL and use this as a test strain suspension.

Add 1 mL of the interfering substance and 1 mL of the test strain suspension, mix immediately, allow to stand in a constant-temperature water bath at 20 ° C for 2 minutes, add 8 mL of the test solution, mix and react in a constant-temperature water bath at 20 ° C for 5 minutes. Take 1 mL of this reaction mixture, place in a test tube containing 8 mL of neutralizing agent and 1 mL of water, and neutralize in a constant-temperature water bath at 20 ° C for 5 minutes. After completion of neutralization, each 1 mL of neutralization reaction solution is added to two Petri dishes, and TSA medium is added to culture. Count the maximum number of colonies in the Petri dish.

The neutralizing agent is a solution containing 3% (V / V) polysorbate 80, 4 g / L sodium lauryl sulfate and 3 g / L lecithin.

The interfering substances are prepared by dissolving the dried yeast extract for microorganisms in water to a concentration of 100 g / L, adjusting the pH to 7.0 ± 0.2 using sodium hydroxide (NaOH), and sterilizing at 121 ° C for 15 minutes.

The reduction rate of viable cell count is calculated according to the calculation formula for each test bacterium and test solution, and sterilization disinfection power is considered to be effective when the reduction rate of viable cell count is 99.999% or more.

≪ Calculation formula of number of living cells of sterilization disinfectant test suspension suspensions >

Number of live cells (cfu / mL) in this test and verification test = c / (nⅹdⅹV)

* c: sum of the number of colonies counted in the Petri dish

* n: Number of counted petri dishes

* d: dilution factor (10 ^{-1 for} dilution of the test strain suspension)

* V: Volume of the sample (1.0 mL in case of dilution of the test strain suspension)

<Antimicrobial Disinfection Test Test>

(%) = (N-10Na) / N ⅹ 100

* N: initial number of bacteria

* Na: Number of live cells (cfu / mL) due to sterilizing action of test solution

* If the number of colonies counted in this test is less than 15, 1.5 × 10 ² cfu / mL of Na is applied, and when the number of counted colonies is more than 300, the number of viable cells (Na) is 3.0 × 10 ³ cfu / mL.

Experimental Example 1. Evaluation of disinfection ability

The sterilizing power of carnoic acid, carnosol, and glabridin was evaluated at concentrations of 100 ppm, 200 ppm, and 500 ppm.

Test results of disinfection of carnosic acid, carnosol and gabridine Applied bacteria Test strain suspension
(cfu / ml) Test substance content
(ppm) Number of live bacteria after sterilization
(cfu / mL) Disinfection power (%) E. coli 2.13x10 ⁸ Carnoic acid 100 > ³ x 10 ³ 99.971 200 <1.5 x 10 ² > 99.999 500 <1.5 x 10 ² > 99.999 Carnosol 100 > ³ x 10 ³ 99.952 200 > ³ x 10 ³ 99.981 500 <1.5 x 10 ² > 99.999 Gabridine 100 > ³ x 10 ³ 99.976 200 > ³ x 10 ³ 99.983 500 <1.5 x 10 ² > 99.999 S. aureus 1.87x10 ⁸ Carnoic acid 100 > ³ x 10 ³ 99.988 200 <1.5 x 10 ² > 99.999 500 <1.5 x 10 ² > 99.999 Carnosol 100 > ³ x 10 ³ 99.913 200 > ³ x 10 ³ 99.975 500 <1.5 x 10 ² > 99.999 Gabridine 100 > ³ x 10 ³ 99.984 200 <1.5 x 10 ² > 99.999 500 <1.5 x 10 ² > 99.999

As shown in Table 1, carnoic acid showed a disinfecting power of 99.999% or more within 5 minutes for both E. coli and S. aureus at a concentration of 200 ppm or more, and showed no specificity for Gram-positive bacteria or Gram-negative bacteria. Carnosol showed more than 99.999% bactericidal disinfection ability at the concentration of 500ppm or more and showed no specificity for Gram-positive bacteria or Gram-negative bacteria.

Glaubridin showed more than 99.999% disinfection ability at over 200 ppm for S. aureus at over 500ppm for E. coli and slightly stronger activity against S. aureus .

In order to verify the distinctive activity of the composition of Example 1 and conventional antibiotics with known antibacterial activity, gentamicin, fucidin, bacitracin, ampicillin, and vancomycin were added at concentrations of 100 ppm, 200 ppm, and 500 ppm The sterilizing power was measured and shown in Table 2.

The results of the sterilization test of gentamicin, fuccidin, bacitracin, ampicillin, and vancomycin Applied bacteria Test strain suspension
(cfu / ml) Test substance content
(ppm) Number of live bacteria after sterilization
(cfu / mL) Disinfection power (%) E. coli 2.13x10 ⁸ Gentamicin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Fusidic acid 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Bashitra Sin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Ampicillin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Vancomycin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC S. aureus 1.87x10 ⁸ Gentamicin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Fusidic acid 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Bashitra Sin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Ampicillin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC Vancomycin 100 > ³ x 10 ³ TNTC 200 > ³ x 10 ³ TNTC 500 > ³ x 10 ³ TNTC

※ TNTC: Too Numerous To Count

As shown in Table 2, the antimicrobial activity of five antibiotics, gentamycin, fucydinic acid, bacitracin, ampicillin, and vancomycin, which are commercially available, was found to be less than 99.999% Respectively.

Taken together, these results indicate that the compositions of Example 1 have a microbicidal mechanism that is different from conventional antibiotics and that they exhibit a significantly shorter microbial killing rate than conventional antibiotics.

This confirms that the compositions of Example 1 can be used as an excellent disinfectant.

Claims (7)

A composition for sterilization and disinfection, which comprises, as an active ingredient, any one or two or more compounds selected from the group consisting of carnosic acid, carnosol, glabridin and salts thereof. The method according to claim 1,
A composition for sterilization and disinfection characterized by comprising carnosic acid or a salt thereof. The method according to claim 1,
Wherein the composition is for sterilizing food, foodstuff, food appliance or animal skin. A food prepared by adding the composition for sterilization and disinfection of claim 1 or 2. A food additive prepared by adding the composition for sterilization and disinfection of claim 1 or 2. An external preparation for skin prepared by adding the composition for sterilizing and disinfecting according to claim 1 or 2. A cosmetic product prepared by adding the composition for sterilization and disinfection of claim 1 or 2.

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