KR20200040984A - Pharmaceutical composition for preventing or treating gastrointestinal disorder comprising as an active ingredient a compound isolated from Bacillus amyloliquefaciens RWL-1 culture medium - Google Patents

Abstract

The present invention relates to a composition for preventing, solving, and treating gastrointestinal diseases comprising a compound isolated from a culture medium of Bacillus amyloliquefaciens RWL-1 strain and is the first to describe the enzyme inhibitory activity of a bioactive compound isolated from Bacillus amyloliquefaciens species. Since the compound of the present invention has excellent urease inhibitory activity, the compound has an effect of inhibiting the urease activity of Helicobacter pylori and making Helicobacter pylori difficult to survive in the gastric mucosa. Therefore, the compound is effective for treating, preventing, and solving gastrointestinal diseases. The compound uses a secondary metabolite of microorganisms and is expected to be useful in the medical and food industries by using it as an important source of a bioactive metabolite with high therapeutic value.

Description

Pharmaceutical composition for preventing or treating gastrointestinal disorder comprising as an active ingredient a compound isolated from Bacillus amyloliquefaciens RWL-1 culture medium}

The present invention relates to a composition for the prevention, amelioration, or treatment of gastrointestinal diseases comprising a compound isolated from a culture of Bacillus amyloliquefaciens RWL-1 strain.

Helicobacter pylori is a spiral-shaped Gram negative bacillus living in the stomach. It is found in epithelial cells among the tissues of the stomach because it is very sensitive to low pH (acidity) and lives in a low-oxygen environment. Colonies are formed between epithelial cells. Helicobacter pylori can also survive in acidic stomachs because it produces an enzyme called urease. Helicobacter pylori causes diseases such as gastritis, gastric ulcer, and duodenal ulcer. More than 90% of patients with duodenal ulcer are infected with Helicobacter pylori, and the removal of these bacteria reduces the recurrence rate of ulcers. In addition, because Helicobacter causes stomach cancer, the International Cancer Institute (IARC) under the International Health Organization (WHO) defines Helicobacter pylori as a first-class carcinogen. Helicobacter infection rates tend to be higher in older countries, such as Asia and Africa, and higher with age.

On the other hand, endophyte (endophyte) refers to a microorganism that has the ability to live in plant tissue and form colonies in the plant's internal tissues without causing obvious disease. Plant endogenous organisms spread throughout all parts of the plant and are isolated from other parts of the plant. They do not damage plants and interact with host plants, but the interaction depends on many factors and can change over time. In this symbiotic relationship, plants provide shelter, protection, and essential nutrients to plant endogenous organisms. Instead, plant endogenous organisms form beneficial interactions with host plants through the regulation of endogenous plant hormones and nutrients, and plants are able to respond to rapidly changing environments. Improves its adaptability. However, phytoendophytes exist in all plant species, but the interaction between phytoendophytes and the host has not been fully studied.

In addition, phytoendophytes show many important biological activities in addition to the interaction between plants and microorganisms. Many bacteria in plant endogenous organisms are known to produce a variety of bioactive and health promoting compounds such as volatile organic compounds, antibiotics, immunosuppressive compounds, anticancer agents, antiviral agents and antifungal agents. In clinical trials, exopolysaccharides with anti-tumor efficacy were found in the first study of Bacillus amyloliquefaciens, which is more than 60% of anticancer drugs, natural products and plant endogenous bacteria. In addition to these anti-tumor substances, plant endogenous species belonging to the genus Bacillus (Bacillus genus) produce a variety of secondary metabolites. For example, Bacillus thuringiensis produces the anti-infective compound anthracene, B. subtilis produces the antibiotic protein YbdN, and these results suggest promising medical possibilities in the genus Bacillus. have.

Separation and identification of biologically active urease inhibitors in recent years is of increasing interest as it can be used as a tool to understand biochemical processes and function as a promising therapeutic agent, although plants are an important source of bioactive metabolites. Microorganisms are also considered to be an important source of bioactive metabolites with high therapeutic value. In this regard, although the biological potential of Bacillus amyloliquefaciens has been widely reported, studies on the enzyme inhibitory activity of bioactive compounds isolated from Bacillus amyloliquefaciens species have been incomplete.

Republic of Korea Patent Registration Publication 10-0673605

The present inventors confirmed the urease inhibitory effect of the crude extract of Bacillus amyloliquefaciens RWL-1 strain culture medium to isolate a compound having a specific structure from the crude extract and experimentally showed that the separated compound has excellent urease inhibitory activity. Confirmed to complete the present invention.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating gastrointestinal disease and a food composition for improving gastrointestinal disease, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

In one embodiment of the present invention, the compound may be isolated from a culture of Bacillus amyloliquefaciens RWL-1 strain (KCTC 13637BP).

In another embodiment of the present invention, the composition may have a urease inhibitory activity effect.

In another embodiment of the present invention, the gastrointestinal disease may be gastritis, gastric ulcer, or gastric cancer caused by Helicobacter pylori.

In addition, the present invention provides a food composition for improving gastrointestinal disease, comprising the compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

In one embodiment of the present invention, the compound may be isolated from a culture of Bacillus amyloliquefaciens RWL-1 strain (KCTC 13637BP).

In another embodiment of the present invention, the composition may have a urease inhibitory activity effect.

In another embodiment of the present invention, the gastrointestinal disease may be gastritis, gastric ulcer, or gastric cancer caused by Helicobacter pylori.

In addition, the present invention provides a method for preventing or treating gastrointestinal diseases, comprising the step of administering to a subject a pharmaceutical composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

In addition, the present invention provides a pharmaceutical composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, for preventing or treating gastrointestinal diseases.

[Formula 1]

The present invention relates to a composition for preventing, improving, and treating gastrointestinal diseases comprising a compound isolated from a culture of Bacillus amyloliquefaciens RWL-1 strain, the composition of the present invention has excellent urease inhibitory activity By having, it has the effect of inhibiting the urease activity of Helicobacter pylori, making Helicobacter pylori difficult to survive in the gastric mucosa. Since it can be used as an important source of biologically active metabolites, it is expected to be useful in the medical and food industries.

1 schematically shows an experimental method for separating a compound of the present invention and confirming its structure.
Figure 2a is a result of experimenting the urease inhibitory activity effect of the crude extract of Bacillus amyloliquefaciens RWL-1 culture.
Figure 2b is the result of the experiment of the cytotoxicity of the crude extract of Bacillus amyloliquefaciens RWL-1 culture medium by MTT assay.
Figure 2c is a result of testing the effect of AChE (acetylcholinesterase) inhibitory activity of crude extract of Bacillus amyloliquefaciens RWL-1 culture.
Figure 3 shows the structure of the compound isolated from the crude extract of Amyloliquefaciens RWL-1 culture.
4 is a result of testing the urease inhibitory activity of the compound isolated from the crude extract.

The present inventors confirmed the urease inhibitory effect of the crude extract of Bacillus amyloliquefaciens RWL-1 strain culture medium to isolate a compound having a specific structure from the crude extract and experimentally confirmed that the separated compound has excellent urease inhibitory activity. The invention was completed.

In an embodiment of the present invention, the crude extract of Bacillus amyloliquefaciens RWL-1 strain was treated to test the urease inhibitory activity, cytotoxicity, and AChE inhibitory activity, and it was confirmed that the crude extract had excellent urease inhibitory activity (( See Example 2).

In another embodiment of the present invention, the compound isolated from the crude extract of Bacillus amyloliquefaciens RWL-1 strain culture was analyzed to confirm its chemical structure (see Example 3).

In another embodiment of the present invention, as a result of evaluating the enzyme inhibitory activity of the isolated compound, it was confirmed that the urease inhibitory activity is excellent (see Example 4).

Therefore, considering the above, the compound isolated from the crude extract has excellent urease inhibitory activity, and thus can be used for the treatment, prevention, or improvement of gastrointestinal diseases.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

As used in the present invention, the term “prevention” refers to all actions that suppress gastrointestinal disease or delay the onset of disease by administration of the pharmaceutical composition according to the present invention.

As used in the present invention, the term “treatment” refers to all actions in which symptoms caused by gastrointestinal diseases are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.

The pharmaceutical composition according to the present invention includes a compound isolated from the culture of Bacillus amyloliquefaciens RWL-1 strain (KCTC 13637BP) as an active ingredient, and may include a pharmaceutically acceptable carrier. . The pharmaceutically acceptable carrier is commonly used in formulation, and includes, but is not limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc. If necessary, it may further contain other conventional additives, such as antioxidants, buffers, if necessary. In addition, diluents, dispersants, surfactants, binders, lubricants, and the like can be additionally added to formulate into injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, the formulations described in Remington's literature can be used to prepare the formulations accordingly. The pharmaceutical composition of the present invention is not particularly limited in the formulation, but may be formulated as an injection or oral intake.

The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously or subcutaneously) according to a desired method, and the dosage may include the patient's condition and weight, the degree of disease, the drug form, It depends on the route and time of administration, but can be appropriately selected by those skilled in the art.

The composition according to the invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, and activity of the drug , Sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the composition according to the present invention may vary depending on the patient's age, sex, and weight, and may be increased or decreased depending on the administration route, disease severity, sex, weight, age, and the like.

As used herein, the term "pharmaceutically acceptable salts thereof" may be prepared by a conventional method in the art, for example, hydrochloric acid, hydrogen bromide, sulfuric acid, sodium hydrogen sulfate, phosphoric acid, carbonic acid Pharmaceuticals with salts with inorganic acids such as or formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestic acid, fumaric acid, lactobionic acid, salicylic acid, or acetylsalicylic acid (aspirin) To form a salt of their acid, or to react with an alkali metal ion such as sodium or potassium to form their metal salt, or to react with an ammonium ion to form another pharmaceutically acceptable salt thereof It means to form.

In the present invention, the compound may be isolated from the culture of Bacillus amyloliquefaciens RWL-1 strain (KCTC 13637BP), but is not limited thereto.

In the present invention, the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) RWL-1 strain was deposited with the Korea Research Institute of Bioscience and Biotechnology Biological Resource Center (KCTC) on September 06, 2018, and was given an accession number KCTC 13637BP.

The strain of the present invention may be isolated from rice seeds, but is not limited thereto.

The composition of the present invention may have an urease inhibitory activity effect.

The term “urease” used in the present invention is also called urease, and is widely distributed in gastric mucus and red blood cells of microorganisms (bacteria, yeast, staphylococcus), higher plants, lower animals, and higher animals. Plants are abundant in legumes, and small beans contain about 0.1% of the dry weight. It is the first crystalline enzyme extracted from J.B. Sumner in 1926 by small beans and is a colorless octahedral crystal with a molecular weight of 483,000, an isoelectric point pH 5.0, and an optimum pH of 8.0. As an active group, it has a -SH group, and enzyme activity is inhibited by heavy metal ions such as mercury, silver, and copper, and oxidizing agents, and is recovered by hydrogen sulfide, cyanate, glutathione, and the like. Bacteria obtain ammonia as a nitrogen source by this enzyme and neutralize hydrochloric acid in the gastric mucosa to protect gastric cells.

In the present invention, the gastrointestinal disease may be gastritis, gastric ulcer, or gastric cancer caused by Helicobacter pylori, but is not limited thereto.

Helicobacter pylori secretes urease, a urea degrading enzyme, to hydrolyze one molecule of urea (H ₂ NCONH ₂ , urea) in gastric juice to form two molecules of ammonia (NH ₃ ). Urease has been closely related to infecting Helicobacter pylori bacteria in human gastrointestinal epidermal cells and assisting in colonization. Specifically, the Helicobacter pylori strain that has inactivated urease is unable to colonize gastric mucosal cells. It has been reported that activity is essential for colonization of Helicobacter pylori, and ammonia produced by Helicobacter pylori urease increases the pH in gastric juice, damages the gastric mucus layer, ammonia itself consumes oxygen in gastric mucosa cells and ATP production in mitochondria , And ultimately ammonia forms monochloroamine to generate reactive oxygen species, causing cell damage to cause chronic inflammation, and further DNA damage to accelerate the cancer-producing process. There are reports.

As another aspect of the present invention, the present invention provides a food composition for improving gastrointestinal disease, comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The food composition includes a health functional food composition.

As used in the present invention, the term "improvement" refers to any action that at least reduces the severity of parameters associated with the condition being treated, such as symptoms.

In the food composition of the present invention, the active ingredient may be added to the food as it is or used with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (for prevention or improvement). Generally, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, with respect to the raw materials in the manufacture of a food or beverage. However, in the case of long-term intake for health and hygiene purposes or for health control purposes, the amount may be below the above range.

The health functional food composition of the present invention is an essential component in the indicated proportions, and there is no particular limitation on other components other than those containing the active ingredient, and may contain various flavoring agents or natural carbohydrates, etc., as additional components, such as ordinary beverages. have. Examples of the natural carbohydrates described above include monosaccharides, for example, glucose, fructose, etc .; Disaccharides such as maltose, sucrose, etc .; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatine, stevia extract (for example, rebaudioside A, glycyrrhizine, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates can be appropriately determined by the choice of those skilled in the art.

In addition to the above, the functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like. These ingredients can be used independently or in combination. The proportions of these additives can also be appropriately selected by those skilled in the art.

Hereinafter, preferred embodiments are provided to help understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Experimental materials and methods

Example  1-1. Bacillus Amyloliquefaciens RWL -1 strain culture and secondary metabolite extraction

The endogenous Bacillus amyloliquefaciens RWL-1 isolated from rice seeds was cultured in 8 liters of LB medium (tryptone, 10 g; yeast extract, 5 g; sodium chloride) ), 10 g; autoclaving for 15 minutes at pH 7.0 ± 0.2, 121 ° C). The inoculated RWL-1 was incubated in a shake incubator (120 rpm) for 7 days at 28 ° C., and then the cells were separated by centrifugation at 5000 × g for 15 minutes, and the culture solution from which the RWL-1 cells were removed was adjusted to pH 2.5. Then, it was completely extracted three times with the same amount of ethyl acetate. Then, the ethyl acetate extract was completely dried on a rotary evaporator to obtain a concentrated extract (1.6 g).

Example 1-2. Separation of secondary metabolites

Based on the bioassay results, silica gel column chromatography using a crude ethyl acetate solvent gradient (1% ethyl acetate / n-hexane to 85% ethyl acetate / n-hexane) was performed. Analysis. Thin layer chromatography (TLC; pre-coated aluminum sheets, silica gel 60F-254, E. Merck, Darmstadt, Germany) experiments were performed to investigate and determine different fractions of bacterial culture extracts. TLC plates were observed under ultraviolet light of 254 nm and 365 nm. For further purification, bioactive fractions were obtained using high performance liquid chromatography (HPLC) combined with a UV-VIS detector (SPD-10A) and Shimadzu CBM-10. Samples were analyzed with a C18 column (Luna 5 μm, 100 mm 2, 250 × 4.6 mm). As the solvent, solvent A-100% methanol (MeOH) and solvent B-water containing 5% acetic acid were used, and a solvent program; 0-20 min (50% = A; 50% = B), It was analyzed using 20-40 min (80% A, 20% B), 40-60 min (100% A, 0% B), flow rate of 1 mL / min. The remaining impurities were removed by loading the semi-refined secondary metabolite in a preparative TLC plate, and 4.5 mg of the compound was purified in DCM / n-hexane (95: 5) (see FIG. 1).

Example 1-3. Confirmation of the structure of the compound separated from the crude extract

Spectral analysis (UV (Ultraviolet Ray), IR (infrared), 1H-NMR (nuclear magnetic resonance), 13C-NMR, electrospray ionization (ESI) and mass spectrometry (MS) / MS for identification and characterization of purified compounds Study). Optical measurements were performed using a polarimeter (JASCO DIP360) and a Bruker ATR-Tensor 37 spectrophotometer was used to record the IR spectrum. A 5-5.5 kV QSTAR mass spectrometer was used to obtain the ESI mass spectrum. NMR spectra (1H and 13C) were obtained using a Bruker NMR spectrometer operating at 600 MHz and 150 MHz, respectively.

Example 1-4. Evaluation of urease inhibitory activity

The effect of urease inhibitory activity of the compound isolated from the ethyl acetate concentrated extract of RWL-1 and the crude extract was investigated. 100 mM urea (0.05 mL) in 8.2 pH phosphate buffer containing 0.01 M LiCl ₂ , 0.1 mM EDTA, 0.01MK ₂ HPO ₄ 3H ₂ O, 3 units / mL jack bean uriase enzyme solution (0.025 mL) and crude extracts at various concentrations were reacted in a 96-well plate at 37 ° C for 15 minutes. The urease inhibitory activity was evaluated by measuring ammonia production by the indophenol blue method. Absorbance was measured at 630 nm, thiourea was used as a standard inhibitor, and enzyme inhibitory activity was calculated through the following equation, and the graph value was expressed as the average of 3 replicates with standard error.

Enzyme inhibitory activity (%) = 100-(OD test / OD control) × 100

Example 1-5. Cytotoxicity evaluation

Cytotoxicity evaluation of the ethyl acetate crude extract and the compound isolated from the crude extract was performed by MTT analysis. HCT-15 cells were purchased from the American Type Culture Collection CCL-25 (USA), and the atmospheric conditions were 95% air, 5% CO ₂ , and humidified subconfluent conditions at 37 ° C. RPM-1640 medium, stored under 10% fetal bovine serum and 1% (v / v) streptomycin, was used for passage. HCT-15 cells were treated with crude acetate extracts of RWL-1 for 24 hours, 48 hours and 72 hours at 10 ⁵ cells / well in 96-well plates at each concentration (250, 500, 750 μg / mL) and untreated. Cultured by density. The medium was removed and 20 μL MTT solution (5 mg / mL in PBS (phosphate buffered saline)) was added to each well of a 96-well plate and incubated at 37 ° C. for 2 hours. After incubation, MTT medium was replaced with dimethylsulfoxide (DMSO; 200 μl).

Then, the plate was gently shaken for 1 minute, the absorbance was measured at 540 nm, and the following equation was used to calculate cell viability, and the graph value was expressed as the average of 3 repetitions with standard error.

Viable cells% = (OD treated sample / OD untreated sample) × 100

Example 1-6. AChE (acetylcholinesterase) inhibitory activity evaluation

AChE inhibitory activity was evaluated based on the Elman's colorimetric method. 15 mM acetylthiocholine iodide-deionized water (25 μL) solution containing 0.1 M sodium chloride (NaCl) and 0.02 M MgCl ₂ 6H ₂ O, 3 mM 5,5-Diti Obis-2-nitrobenzoic acid (5,5-dithiobis-2-nitrobenzoic acid; DTNB, 125 μL), 50 mM Tris-hydrogen chloride buffer (Tris-HCl buffer, pH 8.0) was added to the 96-well plate Did. 50 mM tris-hydrogen chloride buffer (pH 8.0) containing 0.1% bovine serum albumin (50 μL) and ethyl acetate crude extract (25 μL) was added at different concentrations (50 μg / g to 600 μg / g) Did. AChE (25 μL, 0.2 U / mL) was added, absorbance was measured at 412 nm at 45 second intervals, galantamine (0.5-5 μg / mL) was used as a standard inhibitor, and AChE inhibition was performed using the following formula. The graph values were expressed as the average of 3 iterations with standard error.

AChE inhibitory activity (%) = 1-(sample response rate / blank response rate) × 100

Example  2. Bacillus Amyloliquefaciens RWL -One Crude extract  Biological activity assessment

The biological potential of the Bacillus amyloliquefaciens RWL-1 crude extract was evaluated through inhibitory activity and cytotoxicity to various enzymes and treated with various concentrations of the RWL-1 crude extract, according to the method of Example 1-4. The inhibitory activity was investigated for the cytotoxicity of HCT-15 cells by the method of Example 1-5, and the AChE inhibitory activity of the method of Example 1-6.

As a result, as shown in FIG. 2A, the RWL-1 crude extract exhibited a high inhibitory effect of urease as the concentration increased (10-100 μg / mL) and urease (49.4 ±) at a high dose (100 μg / mL). 0.53%), and the positive control showed 90.86 ± 0.08% inhibition.

On the other hand, in the case of cytotoxicity, the RWL-1 crude extract showed a small cytotoxic effect (25 ± 0.16%) at a high concentration (750 μg / mL) compared to the control (100%), as shown in FIG. 2B. AChE inhibitory activity was shown to occur in a dose-dependent manner at a relatively high dose (250-750 μg / mL), as shown in Figure 2c, and AChE compared to the positive control compound significantly inhibited AChE (94.45 ± 0.31%). No significant decrease in activity was observed.

Example 3. Structural analysis of compound isolated from crude extract

The structure of the compound was performed by analysis of NMR and MS spectral data according to the method of Example 1-3 compared to the data reported in the previous study. The 1H-NMR spectrum showed signals for 5 protons in the aromatic region (δ8.76-7.59) due to the benzene and unsaturated γ lactone residues with 3 substituents, in the downfield region of δ9.15. The broad singlet was assigned to the amide proton NH in the molecule. In addition to these aromatic signals, three oxy-methine protons (δ 4.24-3.84) and two methyl groups (6H, d, J = 7.4 Hz) were observed in the 1H-NMR spectrum.

Furthermore, in the 13C-NMR spectrum, the unsaturated γ-lactone appeared in the presence of carbonyl carbon at δ176.2, in addition to the two characteristic sp2 methine signals at δ157.7 and 132.4, and the substituted benzene ring of aromatic methine carbon. δ139.3, 129.9 and 125.3.

In addition, the compound (name: (S) -2-hydroxy-N-((S) -1-((S) -8-hydroxy-1-oxoisochroman-3-yl) -3-methylbutyl ) -2-((S) -5-oxo-2,5-dihydrofuran-2-yl) acetamide, (S) -2-hydroxy-N-((S) -1-((S)- 8-hydroxy-1-oxoisochroman-3-yl) -3-methylbutyl) -2-((S) -5-oxo-2,5-dihydrofuran-2-yl) acetamide) was compared with those reported in the literature. .

As a result, it was found that the overall physical and spectral data of the compound were the same as the antibiotic Al 77F previously isolated from the bacterial strain of Bacillus fumilus and the strain of Alternaria tenus (see FIG. 3).

Example 4. Evaluation of the enzyme inhibitory activity of the compound isolated from the crude extract

Based on the biological potential of the crude RWL-1 extract, the urease inhibitory activity of the compound was tested at various concentrations by the method of Example 1-4.

As a result, as shown in Figure 4, the urease inhibition rate increased as the concentration of the compound (Compound 1) increased from 97.52 μg / mL of the IC50 value, and significant inhibition (51.27 ± 1.0%) when the compound was 100 μg / mL. Showed. On the other hand, the standard drug used as a positive control had IC50 values of 88.24 ± 2.2% at 55.13 μg / mL and 80 μg / mL, and the negative control showed 0.05 ± 0.01% inhibition.

The above description of the present invention is for illustration only, and a person having ordinary knowledge in the technical field to which the present invention pertains can understand that it can be easily modified to other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Depository name: Korea Research Institute of Bioscience and Biotechnology

Accession number: KCTC13637BP

Date of accession: 20180906

Claims (8)

A pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising the compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

According to claim 1,
The compound is characterized in that isolated from the culture of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) RWL-1 strain (KCTC 13637BP), pharmaceutical composition for preventing or treating gastrointestinal diseases.
According to claim 1,
The composition is characterized in that it has an urease (urease) inhibitory activity effect, gastrointestinal disease prevention or treatment pharmaceutical composition.
According to claim 1,
The gastrointestinal diseases are gastritis, gastric ulcer, or gastric cancer caused by Helicobacter pylori, pharmaceutical composition for preventing or treating gastrointestinal diseases.
A food composition for improving gastrointestinal disease, comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

The method of claim 5,
The compound is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) RWL-1 strain (KCTC 13637BP), characterized in that separated from the culture medium, food composition for improving gastrointestinal diseases.
The method of claim 5,
The composition is characterized in that it has a urease (urease) inhibitory activity effect, gastrointestinal disease improvement food composition.
The method of claim 5,
The gastrointestinal disease is gastritis, gastric ulcer or gastric cancer caused by Helicobacter pylori, food composition for improving gastrointestinal disease.

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