KR102504101B1 - Chrysin Derivatives and Antibacterial compositions comprising the Same - Google Patents

Abstract

The present invention relates to a chrysin derivative having antibacterial activity and an antibacterial composition comprising the same, and the chrysin derivative according to an embodiment of the present invention is useful as an antibacterial composition because it has excellent antibacterial activity compared to chrysin. It can be.

Description

Chrysin derivatives having antibacterial activity and antibacterial compositions containing the same

The present invention relates to a chrysine derivative having antibacterial activity and an antibacterial composition comprising the same.

Antibiotics generally refer to antimicrobial agents, in particular, substances that have antibacterial action against bacteria, specifically substances that have excellent antibacterial action by inhibiting the system in which bacteria synthesize cell walls or proteins, or substances manufactured from such substances. it means. The most representative antibiotic is penicillin, which was manufactured by British physician Alexander Fleming in 1928, and a representative antibiotic developed after penicillin is methicillin, which is recognized as more effective than penicillin. Methicillin is produced by partially altering the chemical structure of penicillin.

Antibiotic-resistant bacteria are bacteria that are resistant to certain antibiotics and do not work. For example, there are penicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA), which do not respond to the drug effect of penicillin at all. MRSA has been found to have a unique gene that can show resistance to antibiotics such as penicillin or methicillin. It is reported that MRSA does not infect healthy people and mainly infects patients with weak immunity or patients who have undergone surgery, causing sepsis or pneumonia and causing death. Due to the emergence of such antibiotic-resistant bacteria, interest has been drawn to the development of new antibacterial agents that surpass existing antibiotics.

The present inventors have completed the present invention by synthesizing a chrysin derivative having excellent antibacterial activity against antibiotic-sensitive and antibiotic-resistant microorganisms.

1. Republic of Korea Patent No. 10-1142598

One object of the present invention is to provide an antibacterial composition comprising a compound represented by the following general formula I or a salt thereof as an active ingredient:

[Formula I]

In the above formula, R is substituted or unsubstituted glucose or phosphoric acid.

One aspect of the present invention provides an antibacterial composition comprising a compound represented by the following general formula I or a salt thereof as an active ingredient:

[Formula I]

In the above formula, R is substituted or unsubstituted glucose or phosphoric acid.

According to one embodiment of the present invention, the compound of Formula I may be synthesized from chrysin, for example, it may be synthesized by a bioconversion method by culturing chrysin with an edible microorganism, but is not limited thereto.

According to one embodiment of the present invention, the substituted glucose may be glucose substituted with dicarboxylic acid, preferably glucose substituted with succinic acid.

According to one embodiment of the present invention, the antibacterial composition may have antibacterial activity against microorganisms selected from the genus Staphylococcus or the genus Bacillus . Specifically, the microorganism of the genus Staphylococcus may be methicillin-sensitive Staphylococcus aureus or methicillin-resistant Staphylococcus aureus .

As used herein, the term 'methicillin-resistant Staphylococcus aureus', also called methicillin-resistant Staphylococcus aureus, refers to a bacterium with strong resistance to almost all antibiotics such as penicillin or methicillin. It is the main culprit of hospital infection and is found in large hospitals that use a lot of antibiotics. It penetrates the respiratory system and the affected area of surgical patients and causes serious symptoms such as high fever, chills, pneumonia and low blood pressure.

In the present invention, the antimicrobial composition may have the same meaning as antibiotics, which is a generic term for antimicrobial agents, and may also mean the same as antibacterial agents, bactericides, preservatives, preservatives, or disinfectants, and preferably include microorganisms mentioned below. It refers to substances that can inhibit or inhibit growth and life functions.

The antibacterial composition of the present invention may include the compound of formula I in an amount of 0.001% to 99.99% by weight, preferably 0.1% to 99% by weight, based on the total weight of the composition, and the method of using the antimicrobial composition and Depending on the purpose of use, the content of the active ingredient can be appropriately adjusted.

Meanwhile, the antimicrobial composition may be used as a component of a pharmaceutical composition for preventing or treating a disease requiring an antibacterial effect.

Since the chrysin derivative according to an embodiment of the present invention has excellent antibacterial activity compared to chrysin, it can be usefully used as an antibacterial composition.

1 is a graph showing the results of analyzing the culture products of microorganisms and chrysin by HLPC.
Figure 2 is a graph showing the mass spectrometry results of chrysine derivatives.

Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are intended to illustrate one or more specific examples, and the scope of the present invention is not limited to these examples.

Example One: Chrysin derivative synthesis

Chrysin derivatives were synthesized using the microbial bioconversion method developed by the present inventors.

Pediococcus from the Korean Collection for Type Culture (KCTC), Korea Research Institute of Bioscience and Biotechnology acidilactici (KCTC 3101), Lactobacillus sakei (KCTC 13416) and Bacillus amyloliquefaciens (KCTC 13588) was distributed and cultured for 24 hours in 500 ml of GPY medium (4% glucose, 0.5% peptone and 0.5% yeast extract). After 24 hours, microbial pellets were obtained by centrifugation at 3,000 rpm for 15 minutes.

The pellets of the three obtained microorganisms were mixed, washed with PG buffer (50 mM phosphate pH 7.2 and 2% glycerol), and then suspended in 5 ml of PG buffer. Chrysin was added to the microbial suspension and cultured at 30° C. for 48 hours. After the culture was finished, 3 ml of acetone was added to the cultured product, and vortexing was performed for 3 minutes to suspend the mixture, and then it was allowed to stand at room temperature for 2 hours. Then, the mixed solution of the culture product and acetone was centrifuged to collect the supernatant, the collected supernatant was concentrated and dissolved in 80% methanol, and HPLC (high performance liquid chromatography) was used to determine whether chrysine derivatives were produced. confirmed.

As a result of the confirmation, it was confirmed that three new peaks were formed in addition to chrysin, indicating that the chrysin derivative was synthesized by incubation with microorganisms.

Figure 1 is a graph showing the results of analyzing the culture products of microorganisms and chrysin by HLPC, and it can be seen that three peaks are newly formed in addition to chrysin, and thus chrysin derivatives are synthesized.

Example 2: synthesized Chrysin derivative analysis

The chrysine derivative synthesized in Example 1 was analyzed by LCMS (liquid chromatography-mass spectrometry, liquid chromatography-mass spectrometry).

As a result of the analysis, it was possible to identify three types of derivatives synthesized from chrysine, and the mass of each derivative was also confirmed. The mass spectrometry results are shown in Table 1 below.

mass spectrometry chemical structure chemical formula Chrysin 254.0579

C ₁₅ H ₁₀ O ₄ chrysine derivative 1 334.02

C ₁₅ H ₁₁ O ₇ P chrysine derivative 2 416.11

C ₂₁ H ₂₀ O ₉ chrysine derivative 3 516.13

C ₂₅ H ₂₄ O ₁₂

Figure 2 is a graph showing the mass spectrometry results of chrysine derivatives. The number 1 indicated in the peak means chrysine, and the numbers 2 to 4 represent chrysine derivatives 1 to 3, respectively.

Example 3: Chrysin Confirmation of antibacterial activity of derivatives

The antibacterial activity of the chrysin derivatives against the strains shown in Table 2 below was confirmed through minimum inhibitory concentration (MIC) measurement.

Specifically, 158 μl of MHB medium was dispensed in the first vertical line of a 96-well plate and 80 μl was dispensed in the remaining vertical lines, and 2 μl of a chrysin derivative was added, followed by serial dilution. Next, 20 μl (5x10 ⁵ cfu/ml) of the strains shown in Table 1 below were inoculated into each well and then cultured in a 37° C. incubator for 16 to 20 hours. After the incubation, IC ₅₀ values were measured using an absorbance microplate reader.

As a result of the measurement, as shown in Table 2 below, chrysin has a MIC of 128 μg / ml or more and has little antibacterial activity, but in the case of chrysin derivatives 1 to 3, the MIC is in the range of 0.5 to 128 μg / ml and has antibacterial activity could confirm that In particular, in the case of chrysine derivative 1, it was found that antibacterial activity against antibiotic-resistant microorganisms was remarkably excellent.

MIC (μg/ml) Chrysin Chrysin
derivative 1 Chrysin
derivative 2 Chrysin
derivative 3 S. aureus CCARM 0205 (MSSA) >128 0.5 16 32 S. aureus CCARM 0204 (MSSA) >128 32 >128 >128 S. aureus CCARM 0027 (MSSA) 128 0.5 16 32 S. aureus CCARM 3634 (MRSA) >128 128 128 >128 S. aureus CCARM 3635 (MRSA) >128 One 32 128 S. aureus CCARM 3089 (MRSA) >128 16 >128 >128 S. aureus CCARM 3640 (MRSA) 128 4 64 64 S. aureus ATCC 33591 (MRSA) >128 64 >128 >128 Bacillus subtilis ATCC 6633 >128 16 >128 >128 Kocuria rhizophila NBRC 12708 >128 >128 >128 >128 E. coli ATCC 25922 >128 >128 >128 >128 Salmonella enterica ATCC 14028 >128 128 >128 >128 Enterococcus faecalis 19433 >128 >128 >128 >128

So far, the present invention has been looked at with respect to its preferred embodiments. Those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent scope will be construed as being included in the present invention.

Claims (4)

An antimicrobial composition comprising a compound represented by the following general formula I or a salt thereof as an active ingredient and having antibacterial activity against microorganisms selected from Staphylococcus aureus or Bacillus subtilis:
[Formula I]

In the above formula, R is phosphoric acid.
delete delete The method of claim 1, wherein the Staphylococcus aureus is methicillin-sensitive Staphylococcus aureus or methicillin-resistant Staphylococcus aureus . antibacterial composition.

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