KR20200139412A - An antimicrobial composition comprising zerumbone as active ingredient - Google Patents

Abstract

The present invention relates to an antibacterial composition comprising zerumbone as an active component. Zerumbone, which is an active component of a composition of the present invention exhibits excellent antibacterial activity on Bacteroides fragilis, and inhibits and eliminates the formation of a Bacteroides fragilis biofilm. Therefore, the composition of the present invention can be used for treating, preventing or alleviating intestinal diseases such as acute diarrhea, inflammatory bowel diseases, and colon cancer caused by Bacteroides fragilis.

Description

An antimicrobial composition comprising zerumbone as active ingredient}

The present invention relates to a jerumbone (zerumbone) exhibiting an antibacterial effect, and in detail, to an antibacterial composition against Bacteroides fragilis comprising a jerumbone (zerumbone) as an active ingredient.

Bacteroides fragilis is a normal intestinal bacterium, but is an opportunistic pathogen in ulcerative colitis or inflammatory bowel disease. Many studies have confirmed that Bft, a toxin of Bacteroides fragilis , causes disease (Boleij et al, Clin. Infect. Dis. 60(2015) 208-215). Enterotoxigenic Bacteroides fragilis (ETBF) produces a toxin called bft (fragilysin), which is associated with acute diarrhea, inflammatory bowel disease, and colon cancer. Bacteroides fragilis forms a biofilm on the intestinal mucosa. The biofilm is an extracellular polymeric substance secreted by microorganisms. The polymer complex is a protein, polysaccharide, and nucleic acid. It is made of such materials. Adjacent bacteria aggregate through the biofilm, attach nutrients or prevent microorganisms from falling off. Biofilm serves to protect the host's defense system and bacteria from antibiotics. Thus, the bacteria forming the biofilm have strong viability and are resistant to antibiotics. It is reported that the biofilm density in patients with inflammatory bowel disease is more than twice that of normal people, and the biofilm formed by Bacteroides fragilis accounts for more than 60% of the biofilm density in patients with inflammatory bowel disease. It is known to do. In addition, it is possible to predict the risk of colon cancer by checking the biofilm of the intestinal mucosa. Therefore, inhibition or removal of the biofilm of Bacteroides fragilis may lower the risk of colon cancer as well as inflammatory bowel disease. However, existing studies have shown that it is difficult to cure biofilm infections with only the currently used antibiotic treatment, and various attempts to overcome biofilm infection are urgently needed (Hong et al, Int J Oral Sci. 2015 Mar. ; 7(1): 1-7).

Zerumbone is an extract of Zingiber zerumbet (L.) Smith of the ginger family, and is known to have skin whitening, antibacterial and anti-inflammatory effects (Vishwanatha et al, IJABPT. 2012; Vol 3, Issue4, Kim et al. J. Med. Food 22 (2019) 62-73), the potential for use as a treatment for chronic diseases such as cancer is emerging. However, the effect on Bacteroides fragilis is still unknown.

Boleij et al, Clin. Infect. Dis. 60(2015) 208-215 Hong et al, Int J Oral Sci. 2015 Mar; 7(1): 1-7 Vishwanatha et al, IJABPT. 2012;Vol 3, Issue 4 Kim et al J. Med. Food 22(2019) 62-73

In order to solve the above problems, an object of the present invention is to provide a jerumbon composition exhibiting excellent antibacterial activity against Bacteroides fragilis , and the composition of the present invention is Bacteroides fragilis. ) Can be used as a therapeutic agent for intestinal-related diseases because it has the effect of inhibiting and removing biofilm formation.

In order to achieve the above object, the present invention provides an antimicrobial composition against Bacteroides fragilis containing zerumbone as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a Bacteroides fragilis anti-biofilm composition comprising a zerumbone as an active ingredient.

In order to achieve another object of the present invention, the present invention is made by Bacteroides fragilis comprising a zerumbone or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. It provides a pharmaceutical composition for the treatment or prevention of intestinal diseases.

The composition comprising jerumbon according to the present invention as an active ingredient inhibits the formation of a biofilm of Bacteroides fragilis and exhibits an effect of removing the already formed biofilm, and in particular, is directly related to intestinal diseases. It has excellent effect on enterotoxigenic Bacteroides fragilis ( ETBF), which can treat or prevent related intestinal diseases.

1 is a graph showing the effect of Jerumbon inhibiting the formation of Bacteroides fragilis biofilm (*P<0.05).
2 is a graph and photograph showing the effect of Jerumbon removing the already formed Bacteroides fragilis biofilm (*P<0.05). In the case of FIG. 2B, MBEC ₉₀ of the three strains was the same as 64 ug/mL, but the bacteria did not grow at the corresponding concentration.
3 is a graph showing the effect of Jerumbon on inhibiting the metabolic activity of Bacteroides fragilis biofilm (*P <0.05).
Figure 4 is a CLSM image showing the effect of Jerumbon inhibiting the formation of Bacteroides fragilis biofilm. FIG. 4A is a WT-ETBF ( bft -2), FIG. 4B is a rETBF ( bft -2), and FIG. 4C is a biofilm image of WT- NTBF . In the group treated with 16 ug/mL gerumbone, all three strains showed marked inhibition of biofilm formation.
5 is a photograph and graph showing the anticancer effect of Jerumbon on ETBF/AOM/DSS mice. Figure 5A is a picture of the large intestine, 5B is the number of polyps, 5C is the polyp size, 5D is the large intestine length (mm), 5E is the large intestine weight (mg) / large intestine length (mm) ratio. S is the control group; Z is the zerumbone treatment group, *P <0.05, **P <0.01, ***P <0.001.

In order to achieve the above object, the present invention provides an antimicrobial composition against Bacteroides fragilis containing zerumbone as an active ingredient.

Jerumbone (Zerumbone, CAS Number 471-05-6) is a compound extracted from Zingiber zerumbet (L.) Smith of the ginger family , and has the structure of the following formula (1).

The rumbon of the anti-inflammatory effect and, E. coli (Escherichia coli), Aspergillus duck chair (Aspergillus oryza) The antibacterial effect of rumbon for studies confirm the antibacterial effects, foil teroyi des plastic Gillis (Bacteroides fragilis), but for such Is still unknown.

Jerumbon provided by the present invention exhibits antibacterial activity against Bacteroides fragilis , and in particular, exhibits more excellent antibacterial activity against enterotoxigenic Bacteroides fragilis ( ETBF). Show. The antibacterial activity of Jerumbon is the growth inhibitory effect of Bacteroides fragilis in the suspended state (minimum inhibitory concentration, MIC), which is not attached to the biofilm, and is produced by Bacteroides fragilis . Biofilm inhibitory concentration (BIC), biofilm eradication concentration (BEC), and Bacteroides fragilis growth inhibitory effect after biofilm removal (minimum biofilm eradication) concentration, MBEC) (Table 1).

Therefore, the jerumbon can be used as an antibacterial composition of Bacteroides fragilis .

'Containing as an active ingredient' according to the present invention means that the zerumbone contains an amount sufficient to achieve efficacy or activity.

'Antibacterial' according to the present invention means killing microorganisms or inhibiting growth, or reducing toxicity.

In one specific embodiment, the present invention provides an antimicrobial composition against enterotoxigenic Bacteroides fragilis ( ETBF) comprising jerumbon as an active ingredient.

Enterotoxigenic Bacteroides fragilis (ETBF) is a pro-carcinogenic bacterium that produces a toxin called bft (fragilysin) and is known to be involved in the onset and progression of colon cancer. It is also associated with acute diarrhea and inflammatory bowel disease. ETBF is known to form a biofilm having a density that is more than 10 times higher than that of non-toxigenic Bacteroides fragilis (NTBF). This was also confirmed in Example 6 of the present invention.

In a specific embodiment of the present invention, Jerumbon inhibited the formation of biofilms of both NTBF and ETBF strains (FIG. 1), and showed an effect of removing all biofilms formed by them (FIG. 2). More specifically, the density of the biofilm formed by ETBF is much higher than that of the biofilm formed by NTBF (Fig. 4), and the same concentration of Jerumbon showing the biofilm removal effect in the NTBF strain also removed the same biofilm in the ETBF strain. It showed the effect (MBEC ₉₀ of NTBF and ETBF is the same as 64 μg/ml, Table 1). Considering the difference in the biofilm density of the two strains, it can be seen that jerumbon exhibits a more excellent effect on removing the biofilm of the ETBF strain, and the antibacterial composition containing the gerumbone of the present invention is an enterotoxin producing Bacteroides It shows a more excellent antibacterial effect on enterotoxigenic Bacteroides fragilis ( ETBF).

In another aspect of the present invention, the present invention provides a Bacteroides fragilis anti-biofilm composition comprising a zerumbone as an active ingredient.

The'anti-biofilm' according to the present invention means to suppress the formation of a biofilm generated by microorganisms or to remove a biofilm that has already been formed.

In one specific embodiment, the present invention provides an enterotoxigenic Bacteroides fragilis ( ETBF) anti-biofilm composition comprising jerumbon as an active ingredient.

In another aspect of the present invention, the present invention relates to a bowel disease caused by Bacteroides fragilis comprising a zerumbone or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. It provides a pharmaceutical composition for treatment or prevention.

The'intestinal disease caused by Bacteroides fragilis ' according to the present invention is an intestinal disease caused by Bacteroides fragilis , acute diarrheal disease, ulcerative colitis (ulcerative colitis), Crohn's disease, inflammatory bowel disease (IBD), colorectal cancer, and the like, but are not limited thereto.

In a specific embodiment of the present invention, Jerumbon inhibits the formation of biofilms of NTBF and ETBF strains, and effectively removes biofilms already formed. In addition, the anticancer effect of Jerumbon was also confirmed in an ETBF tumorigenic mouse (ETBF/AOM/DSS mouse) model (Example 7). Therefore, the composition of the present invention can treat or prevent intestinal diseases caused by Bacteroides fragilis .

'Treatment' according to the present invention refers to any action in which symptoms of bowel disease caused by Bacteroides fragilis are improved or beneficially changed by administration of the composition of the present invention.

'Prevention' according to the present invention means any action of inhibiting or delaying the onset of bowel disease caused by Bacteroides fragilis by administration of the composition of the present invention.

The'pharmaceutically acceptable carrier' according to the present invention is commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, Microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. . The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, preservatives, and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

A suitable dosage of the pharmaceutical composition of the present invention can be variously prescribed depending on factors such as the method of formulation, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient. I can. Meanwhile, the oral dosage of the pharmaceutical composition of the present invention is preferably 0.001-100mg/kg (body weight) per day. The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like. It is preferable that the route of administration of the pharmaceutical composition of the present invention is determined according to the type of disease to be applied. The concentration of the active ingredient in the pharmaceutical composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, and the required period, and is not limited to a specific range of concentration.

The pharmaceutical composition of the present invention is prepared in a unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the art. Alternatively, it may be manufactured by placing it in a multi-volume container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

Hereinafter, the present invention will be described in detail with reference to preferred embodiments so that those of ordinary skill in the art can easily implement the present invention. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

Strains, growth conditions and reagents

In this experiment, wild-type enterotoxigenic B. fragilis 86-5443-2-2 (WT-ETBF ( bft- 2)), wild-type non-enterotoxigenic B. fragilis NCTC 93439 (WT-NTBF) and recombinant strain overexpressing bft in Three types of WT-NTBF (rETBF( bft -2)) were used as strains. These strains were purchased from Johns Hopkins University, USA. BHIS supplemented with 1% glucose, hemin, yeast extract and L-cysteine in brain heart infusion (BHI; Difco, Becton-Dickinson and Company, USA) was used as a medium. Bacterial growth was measured by absorbance at 600 nm using a Multiskan GO plate reader (Thermo Fisher Scientific, USA). Jerumbon used in this experiment was purchased from Sigma-Aldrich (St. Louis, MO, USA), and was dissolved in dimethylsulfoxide (DMSO) and used. For the experiment, a zerumbon concentration of 10 mg/ml was used as a stock solution, and the DMSO concentration did not exceed 1% in all assays.

Confirmation of antimicrobial effect (IC ₉₀ ) against Bacteroides fragilis of Jerumbon

In order to measure the MIC (minimum inhibitory concentration, IC ₉₀ ) of Bacteroides fragilis , a medium microdilution method was used. The final concentration of bacteria in BHIS medium was adjusted to 1×10 ⁶ CFU/ml, transferred to a 96-well plate, treated at a concentration of 4 to 64 ug/mL of Jerumbon, and adjusted to a final 200 μl. Incubated at 37° C. for 24 hours in an anoxic state. After cultivation, the growth inhibition of planktonic cells ( B. fragilis not attached to the biofilm) was measured and analyzed by absorbance at 600 nm using a Dultiskan GO plate reader (Thermo Fisher Scientific, USA). MIC (minimum inhibitory concentration, IC ₉₀ ) was defined as the lowest concentration of gerumbone that inhibited bacterial growth by up to 90% compared to the control group (non-gerumbone treatment group).

Absorbance was measured after cell growth, and as a result, Jerumbon inhibited the growth of Bacteroides fragilis strains in a concentration-dependent manner. Specifically, in WT-ETBF ( bft -2) and rETBF ( bft- 2), which have toxins than WT-NTBF, the IC ₉₀ value of Jerumbon was higher, but at concentrations above the IC ₉₀ value, Jeromebon was the growth of these strains. Was completely suppressed (Table 1).

Confirmation of Jerumbon's Bacteroides fragilis inhibitory effect on biofilm formation (BIC ₉₀ )

Crystal-violet biofilm analysis was performed in a 96-well polystyrene plate (BD, Falcon). After inoculating the strain in 8 ml of BHIS medium treated with 4 to 64 ug/mL of zerumbon, the strain was incubated at 37°C for 24 hours in an anoxic state. After incubation, the supernatant was removed, and the wells were washed three times with 200 μl of PBS to remove non-adherent cells. The biofilm in each well was stained with 100 μl of 1% crystal-violet for 5 minutes, washed three times with 350 μl of sterile distilled water, and dried at 60° C. for 1 hour. After drying for 1 hour, 150 μl of 33% acetic acid was added to the well for 20 minutes to dissolve the dye attached to the biofilm. After transferring to a new 96-well plate, absorbance was measured and analyzed at 570 nm using a Multiskan GO plate reader. A sample not treated with gerumbone was used as a control. BIC ₉₀ (biofilm inhibitory concentration) was defined as the lowest concentration of gerumbone that inhibits biofilm formation by up to 90% compared to the control group (non-zerumbone treatment group).

As a result, Jerumbon suppressed the biofilm formation of Bacteroides fragilis in a concentration-dependent manner (FIG. 1). Specifically, BIC ₉₀ values for WT-NTBF was 32ug / mL, WT-ETBF ( bft- 2) and the BIC ₉₀ values for rETBF (bft -2) is 48ug / mL was (Table 1).

Jerumbon's Bacteroides fragilis biofilm removal effect (BEC ₉₀ , MBEC ₉₀ ) confirmed

The evaluation of the concentration of zerumbone required for biofilm removal was performed in a 96-well polystyrene plate (BD, Falcon). The strain was inoculated at a concentration of 1×10 ⁶ CFU/mL in BHIS 8ml medium, and then cultured at 37° C. for 24 hours in an anoxic state. After formation of the biofilm, the medium was removed and washed three times with 200 μl of PBS to remove non-adherent cells. BHIS medium treated with 8-48 ug/mL zerumbon was transferred to each well, and cultured at 37° C. for 24 hours in an anoxic state. After incubation, the supernatant was removed, and the wells were washed three times with 200 μl of PBS to remove non-adherent cells. Crystal-violet biofilm analysis was performed as in Example 3. BEC ₉₀ was defined as the lowest concentration of zerumbon that removes up to 90% of the already formed biofilm compared to the control group (non-gerumbone treatment group).

The MBEC analysis was similarly conducted. After formation of the biofilm, it was washed with PBS, and the 96-well plate was cultured in fresh medium without zerumbon to grow Bacteroides fragilis on the biofilm. Bacteroides fragilis growth was measured absorbance at 600nm. Thereafter, 4 μl of live bacteria were inoculated on a BHIS agar plate and cultured at 37° C. for 24 hours in an anoxic state, and the formed colonies were counted. MBEC ₉₀ (minimum biofilm eradication concentration) was defined as the lowest concentration of gerumbone in which bacteria were not grown by up to 90% after removal of the biofilm compared to the control group (non-gerumbone treatment group).

BEC ₉₀ results were the same at 48 ug/mL in WT-NTBF, WT-ETBF ( bft- 2) and rETBF ( bft- 2) strains. MBCE ₉₀ was also equal to 48 ug/mL in WT-NTBF, WT-ETBF ( bft- 2) and rETBF ( bft- 2) strains (Table 1).

Effect of Jerumbon on the Metabolic Activity of Bacteroides fragilis Already Formed Biofilm

Biofilm metabolic activity of Bacteroides fragilis was determined by XTT [2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide]-reduction assay. Proceeded. The strain was inoculated at a concentration of 1×10 ⁶ CFU/mL in BHIS 8ml medium, and then cultured at 37° C. for 24 hours in an anoxic state. The cultured strain was adjusted to OD600 with 0.5 McFarland, diluted 100 times, dispensed into a 96-well microtiter plate, incubated in an anoxic state at 37°C for 24 hours, and the cultured medium was removed, followed by PBS 200 μl 3 times. Washed. After treatment with Jerumbon and incubation at 37° C. for 24 hours in an anoxic state, the cultured medium was removed, and then PBS 200 μl 3 was washed. The metabolic activity of cells was quantified using the XTT Cell Proliferation Assay Kit (ATCC, Manassas, VA, USA). The frozen XTT reagent and the active reagent were mixed in a ratio of 50:1, and then 50 μl was added to each well, followed by incubation at 37° C. for 3 hours in an anoxic state. Absorbance was measured at 495 nm using a Multiskan GO plate reader, and the percentage compared to the untreated group was calculated.

As a result, Jerumbon suppressed the biofilm metabolic activity in a concentration-dependent manner for all three strains (Fig. 3).

Confirmation of the inhibitory effect of Jerumbon's Bacteroides fragilis biofilm formation through CLSM

Jerumbon's Bacteroides fragilis biofilm removal effect was confirmed by confocal laser scanning microscopy (CLSM). A 24-well glass bottom imaging plate was used in the same manner as in Example 3. After the strain was treated with gerumbone and dispensed on a plate plate, cultured in an anoxic state at 37° C. for 24 hours, the supernatant was removed, and washed three times with 1 ml of PBS. After fixing for 1 hour at room temperature with 3.7% formaldehyde, the fixing solution was removed, and washed three times with 1 ml of PBS. After drying at room temperature for 30 minutes, the biofilm was dyed with a dyeing solution in an amount that could completely cover it. Staining for 1 hour at room temperature with Fluorescein isothiocyanate isomer I (FITC; 10 mg/ml; Sigma-Aldrich, Munich, Germany), Concanavalin A-Alexa Fluor 594 confugate (Con A; 0.1 mg/ml; C-11253, Molecular Probes, Eugene, OR, USA) at room temperature for 30 minutes, 4, 6-diamidino-2-phenylindoldihydrochloride (DAPI; 1 mg/l; Molecular Probes, Eugene, OR, USA) for 45 minutes at room temperature, respectively, followed by PBS Washed 3 times with. After drying at room temperature for 15 minutes, the biofilm was observed using a Zeiss LSM-710 confocal laser microscope (Carl Zeiss, Thornwood, NY, USA). Visualized as images using ZEN software.

As a result, as shown in Figure 4, WT-NTBF, WT-ETBF ( bft -2) and rETBF ( bft -2) treated with 16 ug/mL of Jerumbon were significant compared to the control (non-jerumbon). It has a different biofilm structure. The control group showed a thick biofilm with a dense structure characterized by a large group, but the zerumbone-treated group showed a loose biofilm due to the effect of inhibiting the formation of gerumbone. In addition, it was confirmed that carbohydrates and DNA in the biofilm were reduced after the zerumbon treatment. From these results, it can be seen that Jerumbon inhibits the formation of Bacteroides fragilis biofilm in a concentration-dependent manner.

Comparing the biofilm morphology between WT-NTBF, WT-ETBF ( bft -2) and rETBF ( bft -2), WT-ETBF ( bft -2) and rETBF ( bft -2) strains that produce bft toxins were formed. It can be seen that one biofilm has a higher density than WT-NTBF (Figs. 4(A), 4(B) vs. 4(C)). Looking at the results of the group treated with 16 ug/mL of Jerumbon, biofilm formation was suppressed to a similar level in all three strains, which is the result confirmed in FIG. 1. In addition, looking at the biofilm removal efficacy formed by the WT-ETBF ( bft -2) and rETBF ( bft -2) strains of Jerumbon confirmed in Example 4, similar biofilms at 16 ug/mL of Jerumbon in all three strains. It was confirmed that the removal efficiency was shown (FIG. 2), and the BEC ₉₀ and MBEC ₉₀ values were the same for all three strains (Table 1). In Figure 4, considering that the biofilm of WT-ETBF ( bft -2) and rETBF ( bft -2) exhibits a higher density compared to the WT-NTBF biofilm, similar biofilm formation at the same zerumbon concentration for the three strains From exhibiting inhibitory and elimination efficacy, it can be seen that zerumbon exhibits a more specifically superior effect on toxin-producing ETBF strains.

Confirmation of anticancer effect of Jerumbon in ETBF tumorigenic mouse (ETBF/AOM/DSS mouse) model

In this experiment, wild-type enterotoxigenic B. fragilis 86-5443-2-2 ( bft -2) (sold from Johns Hopkins University, USA) was used, and the ETBF/AOM/DSS mouse model was produced according to the following protocol. . AOM (Azoxymethane, Sigma-Aldrich, St. Louis, Missouri, USA) 10mg/kg was injected once intraperitoneally into 8-week-old female BALB/c mice (purchased from Rion-bio (Korea)). Two days later, mice were ingested with water containing gentamicin (300mg/L) and clindamycin (100mg/L) for 5 days to promote bacterial colonization. After the mice were infected with ETBF, the DSS cycle (DSS for 5 days, distilled water for 16 days) was performed. DSS (dextran sulphate sodium, 36-50 kDa) was purchased from MP Biomedicals (Santa Ana, California, USA). The bacteria were cultured in brain heart infusion broth (Becton, Dickinson and Company, Franklin Lakes, New Jersey, USA), and the number of bacteria was adjusted to 1× ⁹ CFU/200 μl for oral inoculation to mice. Jerumbon was administered simultaneously with DSS treatment. During the two cycles of DSS (9 weeks), Jerumbon (30, 60 mg/kg, po) was administered three times a week. The daily water consumption of each group of mice was observed. After all cycles, the mice were sacrificed and polyp formation in the colon was observed through autopsy.

As a result, zerumbone reduced polyp formation in ETBF/AOM/DSS mice compared to the control group (non-gerumbone treatment group) (FIGS. 5A and 5B). In addition, it was confirmed that the size of the polyp was also reduced (FIG. 5C), from which it can be seen that the growth or progression of the tumor was slowed by the administration of gerumbone. In the group to which gerumbone was administered, it was confirmed that the colon length increased and the colon weight/length ratio decreased compared to the control group (FIGS. 5D and 5E), which can be seen that gerumbone reduced the incidence of tumors.

Claims (8)

Antimicrobial composition against Bacteroides fragilis containing jerumbone (zerumbone) as an active ingredient. The method of claim 1,
The antimicrobial composition that the Bacteroides fragilis ( Bacteroides fragilis ) is enterotoxigenic Bacteroides fragilis (enterotoxigenic Bacteroides fragilis, ETBF). Bacteroides fragilis containing jerumbone (zerumbone) as an active ingredient ( Bacteroides fragilis ) anti-biofilm (biofilm) composition. The method of claim 3,
The Bacteroides fragilis ( Bacteroides fragilis ) is an enterotoxigenic Bacteroides fragilis (enterotoxigenic Bacteroides fragilis, ETBF) that will be an anti-biofilm (biofilm) composition. Jerumbone (zerumbone) or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition for the treatment or prevention of intestinal diseases by Bacteroides fragilis containing a pharmaceutically acceptable carrier. The method of claim 5,
The pharmaceutical composition of the Bacteroides fragilis ( Bacteroides fragilis ) is enterotoxigenic Bacteroides fragilis (enterotoxigenic Bacteroides fragilis, ETBF). The method of claim 5,
The pharmaceutical composition is Bacteroides fragilis ( Bacteroides fragilis ) biofilm (biofilm) production inhibition or the resulting biofilm is a pharmaceutical composition achieved by. The method of claim 5,
The intestinal disease is acute diarrheal disease, ulcerative colitis, Crohn's disease, inflammatory bowel disease (IBD) or colorectal cancer.

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