KR0131288B1 - Aerosol spray composition for removing fungus using new fungicides - Google Patents

Abstract

Here is provided an aerosol spray composition for eradicating mildew -to remove mildew from its habitat or inhabitable place by convenient spray. An antifungal AF-021A of molecular formula C52H85022N11 and molecular weight 1215, composed of glypeptide antibiotics, includes betahydroxyl asparagine, aspartic acid, serine, glycine, 2,4-diamino butyric acid, betahydroxyl tyrosine, methylene chain compound and gyrase at the ratio of 1:1:2:1:1:1:1:1.

Description

Aerosol Spray Compositions for Mold Removal Using New Antifungal Agents

The present invention relates to an aerosol spray composition containing a new antifungal agent by spraying in the mold, that is, the vehicle air conditioner, bathroom, damp indoor wall, carpet, etc. to give excellent mold removal effect.

Fungi are not only non-pathogenic bacteria that are generally known, but also a lot of deadly bacteria. Representative diseases caused by these pathogenic fungi are bronchial fungal disease and fungal pneumonia caused by infection with Aspergillus fumigatus.

In addition to bronchial fungal disease, fungal diseases include acute sinusitis and skin diseases (USP No. 4,950,477). Bronchococcosis is very difficult to treat once an infection has developed. Conventionally, amphotericin-B (Amphotericin-B) injection and surgery have been used as a therapeutic means for the treatment of bronchial aseptic disease. However, amphotericin-B is very painful and difficult for patients to endure. It is reported that a large amount, such as, elongation.

Diseases caused by fungal infections are mostly caused by inhalation of mold spores parasitic in living and living environment, and prevention of such diseases is made possible by removing parasitic fungi around. An easy way to get rid of these fungi is to use aerosol sprays to spray them where they live or where they live, potentially eliminating mold in the surrounding environment.

The following requirements must be met to make an aerosol spray type mold removal composition.

1) As a small amount of antifungal agent, it should have great antifungal activity.

2) There should be no strong irritation to skin and eyes.

3) Environmental residual activity should be strong.

The present inventors have developed and filed an anti-fungal regulation AF-021A that satisfies all these items (Korean Patent Application No. 93-22090), and this AF-021A is a glycopeptide antibiotics having a molecular weight of 1215 and represented by the molecular formula of C ₅₂ H ₈₅ O ₂₂ N ₁₁ , betahydroxy asparagine, aspartic acid, serine, glycine, 2,4-diamino butyric acid (DAB), betahydroxy tyrosine, methylene chain structure Amino acid and Xylose are composed of 1: 1: 2: 1: 1: 1: 1: 1, and the amino acid binding sequence from the N-terminus to the C-terminus of the peptide is serine-betahydrate. Roxy tyrosine-aspartic acid-serine-glycine-2,4-diaminobutyric acid-betahydroxy asparagine-methylene chain structure Amino acid sequence, and xylose is a cyclic peptide bound to methylene chain tank amino acid .

The details of the antifungal agent AF-021A are well specified in Korean Patent Application No. 93-22090.

The above anti-development AF-021A developed by the present inventors has two kinds of AF-021A ₁ and AF-021A ₂ (hereinafter, collectively referred to as 021A), and cultured Pseudomonas cepacia KFCC 10773 as a microbial origin. It was produced, it was found that the minimum inhibitory concentration (MIC) value is superior to the existing amphotericin-B, ketoconazole and is suitable antifungal agent for use in the present invention.

In the present invention, the above antifungal agent AF-021A is dissolved in a solvent such as ethanol or isopropyl alcohol to prepare an aerosol spray composition. In this case, additives commonly used in spray-type compositions may be used, if necessary.

Hereinafter, the present invention will be described in detail with reference to Examples and Examples.

Reference Example 1.

MIC comparison of antifungal agent and AF-021A.

Yeast is first cultured in a liquid medium, and molds such as Aspergillus are spread on potato dextrose agar (PDA), and then spores are collected and used for testing. It was. The MIC test used the Agar plate method, not the dilution method.

Agar platelets were prepared by adding 0, 1, 10, 15, 20, 25 μg / ml of amphotericin-B, ketoconazole and AF-021A to PDA. The spores and bacteria prepared above were plated on the medium to determine the range where the bacteria did not grow first, and then the concentration range was narrowed again to determine the concentration at which the respective bacteria did not grow. The results are shown in Table 1 below.

As shown in Table 1, it was found that AF-021A was superior to conventional antifungal agents in the MIC value. Therefore, if AF-021A was used to make a mold remover, it could have a greater effect at a lower concentration than conventional products. I could confirm it.

Reference Example 2. Skin irritation test of AF-021A

The anti-fungal agents of amphotericin-B, ketoconazole and AF-021A used in the present invention were investigated for irritation to the skin. Human skin patch test was used as a test method. First of all, 15 subjects were selected from 20 to 30 years old regardless of female and male subjects.

Dissolve each antifungal agent to 200μg / ml, absorb each solution in paper disc (diameter 0.8mm), and put it in a white chamber (Finn chamber: Norgesplaster A / S, Norway) in the lower part of the arm of the subject. It adhered for time and determined on the following criteria.

[Criteria]

++: In case of severe redness (2 points)

+: If there is some redness (1 point)

±: When redness is weak (0.5 point)

-: No redness (0 points)

The test results are shown in Table 2 below.

As shown in Table 2, AF-021A was relatively less irritating to the skin than other conventional antifungal agents, and was found to be suitable for aerosol spray production.

Reference Example 3. Eye irritation test of AF-021A

Tests on eye irritation were carried out by concentration (100, 150, 200, 250 ppm) before and after the same concentration (200 μg / ml) as in the skin irritation test of Reference Example 2, and the test method was an Eyetex system (Eytex (Ropak). G) system) was performed using the Eytex reagent UMA, and finally determined using the software provided by Ropak (Softeware). Criteria are shown in Table 3.

As shown in Table 4, AF-021A was determined to be the minimum stimulus below 220ppm, and amphotericin-B and ketoconazole were judged to be slight stimulation, so that AF-021A was much less irritating to eyes than conventional antifungal agents. It can be seen.

Example. Preparation of Aerosol Spray Using AF-021A and Its Performance Test.

An aerosol spray was prepared according to the composition of Table 5 using AF-021A.

Each unit of Table 5 was added in order to dissolve well, and then put into a 250 ml can, and filled with 70 g of liquid propane gas (hereinafter referred to as LPG) to prepare an aerosol spray.

In addition, a comparative experiment was conducted by dissolving the same composition in ethanol instead of isopropyl alcohol.

First, disperse Aspergillus spores on PDA medium with 0.1% Tween 20 solution. Dilute to cell / ml. The spores were washed with the same solution and washed twice with distilled water. 10 with distilled water again After diluting to cell / ml, Whatman filter paper No. 2 was cut into circles 9 cm in diameter, and 3 ml of distilled water was absorbed therein, and 1 ml of the spore solution was evenly distributed again. The filter paper was attached to a disposable petri dish, and the prepared aerosol spray was sprayed once and incubated at 28 ° C. for 2 weeks to investigate whether or not the spores were killed. The results are shown in Table 6 below.

As shown in Table 6 above, when the solvent is isopropyl alcohol and ethanol, the mortality rate is different and it can be seen that more than 99% sterilization in ethanol.

Reference Example 4. Stability Test of AF-021A

The following items were focused on the stability of the AF-021A in the environment.

(1) In order to investigate the stability of AF-021A at acidic pH and alkaline pH, AF-021A was added to 0.2M Citrate-Phosphate buffer (pH 3, 5, 7, 9, 10) at 100 ppm, respectively, at 30 ° C. Antimicrobial activity was examined at intervals of 2, 4, 6 and 8 weeks. The antimicrobial activity test used the method of the Example. As a result, after 8 weeks, 80% of the activity was maintained in acid (pH 4), and at least 95% of the activity remained at alkaline pH.

(2) In order to find out the high temperature stability of AF-021A, it was tested by dissolving 100ppm in 50% ethanol (pH7), and the antimicrobial activity was maintained even when stored at 60 ℃ for 1 day. Activity was reduced by 50%.

(3) Stability over time in LPG gas filled state

When the aerosol spray was prepared according to the composition of the example, the pressure in the can was about 5-7 kgf / cm 2. How stable the AF-021A under such pressure was examined at one month intervals. The antimicrobial activity test used the method of the Example. As a result, there was no decrease in antimicrobial activity for 14 months when the pressure was below 7kgf / ㎠. As described above, AF-011A maintained high stability at high temperature, high pressure, acidic and alkaline pH, and was found to be the best antifungal agent for aerosol spray.

Claims (2)

It has a molecular weight of 1215 and is represented by a molecular formula of C ₅₂ H ₈₅ O ₂₂ N ₁₁ , betahydroxy asparagine, aspartic acid, serine, glycine, 2,4-diamino butyric acid, betahydroxy tyrosine, methylene chain amino acid Xylose is composed of 1: 1: 2: 1: 1: 1: 1: 1, and the amino acid binding sequence of the peptide from the N-terminus to the C-terminus is serine-betahydroxytyrosine-aspar. Tic acid-serine-glycine-2,4diaminobutyric acid-betahydroxy asparagine-methylene chain amino acid sequence, containing antifungal agent AF-021A which is a cyclic peptide in which xylose is bound to methylene chain amino acid Aerosol Spray Composition For Removing Mold. The aerosol spray composition of claim 1 wherein the concentration of AF-021A is 100-200 ppm.