KR101211669B1 - Antimicrobial compounds from Scilla scilloides and composition containing thereof - Google Patents

Abstract

The present invention relates to a silacylside compound extracted from Scilla scilloides and exhibiting antimicrobial activity, and an antimicrobial composition comprising the compound.
The silacylside-based compound of the present invention and the antimicrobial composition comprising the same are Aspergillus spp., Colletotrichum spp., Pyricularia spp., Fusarium genus ( Fusarium spp.) Or Candida sp. It shows excellent antibacterial effect against various microorganisms including fungi, and because it is extracted from natural products, it is less concerned about side effects. And may be widely used for prevention.

Description

 Antimicrobial compounds and antimicrobial compositions comprising the same

The present invention relates to a silascilloside-based compound extracted from Scilla scilloides and exhibiting antimicrobial activity, and an antimicrobial composition comprising the compound.

Scilla scilloides , one of Korean native plant species, is known as cotton bud and is known to have strong heart -like action, such as digitaris. Aqueous extracts of rhizome alcohol showed significant cardiac effect on toad extraction heart, increased contractile force of heart, increased tension of myocardium and stopped systolic after treatment, but no significant effect on heart rate. Leaf extract action is stronger than the root and significant diuretic activity has been reported in anesthetized dogs.

In particular, it has been used for edible in civilian, and its toxicity to human body has not been reported.

On the other hand, Korean Patent Laid-Open Publication No. 1999-48808 discloses a subterranean section of a dry plant, which is extracted with a mixed solution of water and methanol and concentrated. It has been disclosed that the extract obtained by washing and fractioning exhibits antioxidant activity.

However, no studies have yet been made regarding the antimicrobial activity of the extract.

The present inventors have completed the present invention by discovering that the silacylside-based compound extracted from the plant has antibacterial activity against a specific microorganism.

Therefore, an object of the present invention is to provide the silacylside-based compound exhibiting antibacterial activity.

In addition, another object of the present invention is to provide an antimicrobial composition comprising the silacyl side-based compound.

The present invention is represented by the following formula (1), characterized by a compound exhibiting antimicrobial activity.

In Formula 1, R ₁ is H or OH, R ₂ is H or OCOCH ₃ , R ₃ is rhamnopynosyl or arabinofuranosyl, R ₄ is glucopyranosyl or glucopyra Glucopyranosyl-galactopyranosyl-glucopyranosyl.

In addition, the present invention is characterized by an antimicrobial composition comprising the compound.

Eusterol oligosaccharide compounds of the present invention and antimicrobial compositions comprising the same are Aspergillus spp., Colletotrichum spp., Pyricularia spp., Fusarium genus ( Fusarium spp.) Or Candida sp. Shows excellent antibacterial effect against various microorganisms including fungi, and since it is extracted from natural products, there is little concern of side effects, animal diseases, plant diseases, etc. It can be widely used for the treatment and prevention of

1 is a photograph showing that the silacylside compounds (E-2, E-3) of the present invention exhibit inhibitory activity on the medium against Aspergillus flavers (left) and Chlorella regularis (right).

Hereinafter, the present invention will be described in detail.

The present invention is characterized by a compound represented by the following formula (1) and exhibiting antimicrobial activity.

[Formula 1]

In Formula 1, R ₁ is H or OH, R ₂ is H or CH ₃ COO, R ₃ is rhamnopynosyl or arabinofuranosyl, R ₄ is glucopyranosyl or glucopyranosyl Pyranosyl (glucopyranosyl)-galactopyranosyl-glucopyranosyl.

Further, more preferably in the compound of Formula 1,

R ₁ is H, R ₂ is H, R ₃ is α-L-ramnopyranosyl and R ₄ is β-D-glucopyranosyl; R ₁ is OH, R ₂ is H, R ₃ is α-L-arabinofuranosyl and R ₄ is β-D-glucopyranosyl; R ₁ is H, R ₂ is CH ₃ COO, R ₃ is α-L-lamnopyranosyl and R ₄ is β-D-glucopyranosyl; And R ₁ is H, R ₂ is H, R ₃ is α-L-lamnopyranosyl and R ₄ is β-D-glucopyranosyl- ³ β-D-galactopyranosyl- ³ β-D-glucopyra It is characterized by a compound selected from the group consisting of compounds which are nosyl.

The compound may be derived from any one, and may be obtained by appropriately combining known methods used for separation and extraction of plant components, such as extraction by organic solvents such as alcohol, ether, ketone, column chromatography, Modifications of this are possible within the scope of the known art generally applicable in the art. In addition, the crude extract may be purified by repeating as many times as necessary.

The chromatography used in the present invention may be applied to silica gel column chromatography, LH-20 column chromatography, thin layer chromatography (TLC), high performance liquid chromatography, and the like. Can be.

The compound of the present invention has an antimicrobial effect against various pathogenic microorganisms, in particular Aspergillus sp., Colletotrichum spp., Pyricularia spp., Fusarium genus ( Fusarium spp.) Or Candida sp. Fungi have excellent antimicrobial effects, and also have a killing effect on algae such as Chlorella sp.

In addition, the present invention includes the extract of the extract with a C ₁ ~ C ₆ alcohol to the antibacterial activity of the extract as a right range, wherein the alcohol is preferably used methanol or ethanol.

In addition, the present invention includes the compound represented by the formula (1) or an antimicrobial composition comprising the root extract as a right range.

The antimicrobial composition of the present invention may contain 0.001 to 99% by weight of the euosterol oligosaccharide compound represented by the formula (1) alone or an additive appropriately selected according to the purpose of use. The present invention is not particularly limited to the selective use and content of these additives, and can be appropriately adjusted according to the method of use and the dosage form at the expert level in the art. That is, the additive of the present invention may be a material included in a conventional antibacterial agent, for example, an excipient or a diluent, and examples thereof include saline solution, buffer solution, dextrose, isoparaffin, water, glycerol, and ethanol. It is by no means limited.

The formulation of the antimicrobial agent can be appropriately adjusted according to the method of use and use, such as liquids, suspensions, granules, fragrances, powders, syrups, aerosols, extracts, ointments, derived extracts, emulsions, whole preparations, eye drops, injections, It may be a pill, a capsule, a cream, or a pill.

The antimicrobial composition of the present invention is used for inhibiting the production and growth of human pathogenic and phytopathogenic fungi, and can be used as a detergent, a pharmaceutical, a food additive, a fragrance, a cosmetic, and a biologic. When used as a cleaning agent, it can be prepared as a spray, cleaning agent, shampoo, rinse, soap, etc., wherein the antimicrobial composition may include 0.01 to 50% by weight, but is not limited thereto. When used as a medicament can be used as a disinfectant, therapeutic agent, prophylactic agent, etc. At this time, the dosage of the antimicrobial composition may be 1 to 1000 mg per day, but is not limited thereto. When used as a food additive, the antimicrobial composition is preferably adjusted appropriately according to the type of food and the method of use, for example, it can be used in 0.0001 to 10% by weight relative to the food weight. When used as a fragrance, it may further include a volatile substance, the antimicrobial composition may be used in 0.0001 to 90% by weight. When used as a cosmetic, the antimicrobial composition may be included in 0.0001 to 30% by weight.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments of the present invention, which is intended to illustrate the present invention, but the present invention is not limited thereto.

Example 1 Isolation and Purification of the Silacylside Compound

Bulbs were harvested and dried, and 2.5 kg of bulbs were dried and immersed in 5 L of methanol for 7 days, and extracted three times at room temperature. The methanol extract obtained through the extraction process was suspended in 1 L of water, and the solvent was fractionated twice with 1 L of ethyl acetate and 3 times with 1 L of saturated n -butanol. 10 g of the n -butanol fraction was gel filtered through Sephadex LH 20 (3.0 ㅧ 70 cm, 210 g) using methanol as an elution buffer to obtain 20 fractions (Fr.1 to Fr. 20), each 100 ml. Aqueous methanol solution (75%, 1.5 ml / min, det. At 210 nm) was flowed into YMC-Pak ODS-AQ column (300 ㅧ 10 mm ID, 120A) from elution buffer from 6 g of fractions 3 to 5 of the fraction. First, preparative-HPLC was performed to obtain fifteen subfractions (Fr. 1 'to Fr. 15') of 50 ml each. The sub fraction was further subjected to preparative-HPLC while flowing 75% aqueous methanol solution. Compound 1 (100 mg) was obtained from Fr.5 'and Fr.6', and Compound 2 from Fr.8 '. (18 mg), Fr. 10 ', 11' from compound 3 (12 mg), Fr. Compound 4 (14 mg) was obtained from 11 '.

Example 2 Characterization and Structure Analysis of the Extracted Compound

Melting point (Electrothermal instrument, Dubuque, IA, USA), optical rotational force (DIP-370, Digital Polarimeter, JASCO) were measured to analyze the properties of the compounds 1 to 4, UV spectra (Milton Roy 3000 spectrometer, Ivyland, PA, USA). In addition, the molecular weight was measured by ESI mass spectrometer (JMS 700 mass spectrometer, JEOL, Tokyo, Japan) to analyze the structure of the compound.

(1) Silacylside E-1 (Compound 1)

Shape: white amorphous powder

Melting Point: 221-223 ℃

[α] _D ²⁵ : -57.1 ° (c0.08, MeOH)

MW: 1222

ESI-MS m / z: 1245.6 [M + Na] ⁺ , 1221.7 [MH] ⁻

(2) Silacylside E-2 (Compound 2)

Shape: white amorphous powder

Melting Point: 210-216 ℃

[α] _D ²⁵ : -38.1 ° (c0.08, MeOH)

MW: 1224

ESI-MS m / z: 1247.7 [M + Na] ⁺ , 1223.7 [MH] ⁻

(3) Silacylside E-3 (Compound 3)

Shape: white amorphous powder

Melting Point: 218-221 ℃

[α] _D ²⁵ : -51.9 ° (c0.08, MeOH)

MW: 1220

ESI-MS m / z: 1303.6 [M + Na] ⁺ , 1219.6 [MH] ⁻

(4) Silacylside G-1 (Compound 4)

Shape: white amorphous powder

Melting Point: 240-245 ℃

[α] _D ²⁵ : -46.8 ° (c0.08, MeOH)

MW: 1547

ESI-MS m / z: 1570.2 [M + Na] ⁺ , 1546.2 [MH] ⁻

Compounds 1 to 4 of the silacylside form have the structure of Formula 1 (Table 1).

[Formula 1]

R ₁ R ₂ R ₃ R ₄ Compound 1 H H rha glc Compound 2 OH H ara glc Compound 3 H Oac rha glc Compound 4 H H rha glc- ³ gal- ³ glc rha: α-L-rhamnopyranosyl, ara: α-L-arabinofuranosyl,
glc: β-D-glucopyranosyl, gal: β-D-galactopyranosyl.

Example  3: Determination of antimicrobial activity of compounds 1 to 4

Bacillus subtilis KCTC 1914, Escherichia coli KCTC 1924, Salmonella typhimurium KCTC 1926, Staphylococcus to measure the antimicrobial activity of the extracted compounds 1-4 Aureus ( Staphylococcus aureus KCTC 1916, KCTC 1928), Streptococcus mutans DSM 6178), Aspergillus flavus EML-AF01), Chlorella regyulra less (Chlorella regularis EML-CR02) and Candida albicans KCTC 1940 were measured for antimicrobial activity by paper disc method.

The Bacillus subtilis and Staphylococcus aureus were incubated in Nutrient agar medium (peptone 5 g / l, meat extract 3 g / l, agar 15 g / l) at pH 7.0 and Candida albican Was incubated in YMPG agar medium (3 g / l yeast extract, 3 g / l malt extract, 5 g / l soybean extract, 10 g / l glucose, 15 g / l agar), and Aspergillus Pulabus agar It was incubated in medium (200 ml / l potato extract, 20 g / l dextrose, 15 g / l agar). In addition, Chlorella regularus is Arnon's A5 medium (KH ₂ PO ₄ 1 g / l, MgSO ₄ ~ 7H ₂ 0 1 g / l, FeSO ₄ ~ 7H ₂ 0 0.005 g / l, yeast extract 5 g / l, glucose 20 g / l, agar 20 g / l).

After dissolving the extracted compounds 1 to 4 in methanol and dropping 250 μg each on a paper disk (Whatman NO. 1, 8 mm diameter), the strain was placed on a 200 μl smeared agar plate and completely dried for 24 hours. Then, the antimicrobial activity was confirmed by measuring the size of the inhibitory zone (inhibitory zone), and the results are shown in Table 2 and FIG. 1.

Strain Inhibition zone (mm) Compound 1 Compound 2 Compound 3 Compound 4 Escherichia coli KCTC 1924 - - S - Salmonella typhimurium KCTC 1926 - - - - Staphylococcus aureus KCTC 1916 - - - - Staphylococcus aureus KCTC 1928 - - - - Bacillus subtilis KCTC 1914 - - - - Chlorella regularis EML - CR02 12 11 14 13 Aspergillus flavus EML-AF01 21 16 22 15 Candida albicans KCTC 1940 15 11 16 10 -: No activity, S: weak activity

In addition, anthrax pathogens ( Colletotrichum acutatum EML-CA1), wilted pathogens ( Fusarium oxysporum EML-WIL1), bacillus pathogens ( Pyricularia Oryza e EML-PO1), such as phytopathogens were examined for inhibitory activity by replacement culture method, the results are shown in Table 3 below.

Strain Replacement Culture Effect of Extracts Anthrax pathogen ( Colletotrichum acutatum EML-CA1) ++ Fusarium oxysporum EML-WIL1 ++ Pyricularia oryza e EML-PO1 +++ +++: very active, ++: active steel,-: no active

Through the above experiments, it was confirmed that the silacylside compounds 1 to 4 showed excellent antibacterial activity against fungal species and algae including Aspergillus, and in particular, the compound 3 showed the best antifungal effect. Confirmed. However, the compound did not show antimicrobial activity against gram positive and negative bacteria.

Claims (8)

A compound represented by the following Chemical Formula 1 and showing antibacterial activity:
[Formula 1]

In Formula 1, R ₁ is H or OH, R ₂ is H or CH ₃ COO, R ₃ is rhamnopynosyl or arabinofuranosyl, R ₄ is glucopyranosyl or glucopyranosyl Pyranosyl (glucopyranosyl)-galactopyranosyl-glucopyranosyl.
The method of claim 1,
R ₁ is H, R ₂ is H, R ₃ is α-L-ramnopyranosyl and R ₄ is β-D-glucopyranosyl;
R ₁ is OH, R ₂ is H, R ₃ is α-L-arabinofuranosyl and R ₄ is β-D-glucopyranosyl;
R ₁ is H, R ₂ is CH ₃ COO, R ₃ is α-L-lamnopyranosyl and R ₄ is β-D-glucopyranosyl; And
R ₁ is H, R ₂ is H, R ₃ is α-L-lamnopyranosyl and R ₄ is β-D-glucopyranosyl- ³ β-D-galactopyranosyl- ³ β-D-glucopyranosyl A compound selected from the group consisting of phosphorus compounds.
The compound of claim 1 or 2, wherein the compound is derived from Scilla scilloide .
4. The compound of claim 3, wherein the compound is isolated from any C ₁ to C ₆ alcohol extract.
The genus Aspergillus spp., Colletotrichum spp., Pyricularia sp., Fusarium spp. Or) Candida sp. A compound characterized by having an antimicrobial activity against fungi.
delete An antimicrobial composition comprising the compound of claim 1 as an active ingredient.
8. The composition of claim 7, wherein the composition is of the genus Aspergillus sp., Colletotrichum spp., Pyricularia spp., Fusarium spp. Or Candida. Antimicrobial composition, characterized in that it has an antimicrobial activity against the genus Candida sp.

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