Patent Application
20250127828
This application claims the benefit of priority from the prior Korean Patent Application No. 10-2023-0142894, filed on Oct. 24, 2023, the disclosures of which are incorporated herein by reference in its entirety.
The entire contents of the electronic file identified by its name Sequence Listing.xml, creation date Aug. 20, 2024, and size 3,593 bytes are incorporated herein by reference. Neither the presence nor absence of information that is not required under the sequence rules will create a presumption that such information is necessary to satisfy any of the requirements of 35 U.S.C. 112. Further, the grant of a patent on an application that is subject to the sequence rules (37 CFR 1.831 et seq.) constitutes a presumption that the granted patent complies with the requirements of these rules.
The present disclosure relates to a novel Bacillus velezensis strain and a method for preventing, treating, or improving tuberculosis including the same. More specifically, the present disclosure relates to a Bacillus velezensis PMC 206 strain extracted from vinegar and a use thereof, for example, an antibacterial composition having antibacterial activity against bacteria belonging to the genus Mycobacterium, and a pharmaceutical composition or food composition for improving, treating, or preventing tuberculosis having antibacterial activity against bacteria belonging to the genus Mycobacterium.
Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis. Tuberculosis remains a major global public health problem, causing about 1.8 million deaths each year. Drug-resistant Mycobacterium tuberculosis, including multi-drug resistant (MDR) tuberculosis, broad-spectrum resistant (XDR) tuberculosis, and totally drug resistant (TDR) tuberculosis, continues to pose a threat to public health. Conventional antibiotics are less effective when they are used to treat antibiotic-resistant tuberculosis. Additionally, in the case of existing therapeutic agents, side effects due to long-term high doses of the drug cause serious problems.
Accordingly, in order to overcome these problems, there is an urgent need to develop a new type of anti-tuberculosis drug that is different from the existing compound-based antibiotics.
An object of the present disclosure is to provide a Bacillus velezensis PMC 206 strain.
Another object of the present disclosure is to provide an antibacterial composition including a Bacillus velezensis PMC 206 strain.
Still another object of the present disclosure is to provide a composition for preventing, treating, or improving tuberculosis including a Bacillus velezensis PMC 206 strain.
According to the first exemplary embodiment, the present disclosure provides a Bacillus velezensis PMC 206 strain or a culture thereof, which has antibacterial activity against bacteria belonging to the genus Mycobacterium.
According to the present disclosure, the Bacillus velezensis PMC 206 strain may be extracted from vinegar. For example, Bacillus velezensis PMC 206 strain is assigned Accession No. KCTC 15187BP.
According to the present disclosure, the bacteria belonging to the genus Mycobacterium may be one or more selected from the group consisting of M. abscessus, M. africanum, M. asiaticum, M. avium complex (MAC), M. bovis, M. chelonae, M. fortuitum, M. gordonae, M. haemophiluscellulare, M. kansasii, M. lentiflavum, M. leprae, M. malmoense, M. marinum, M. microti, M. phlei, M. scrofulaceum, M. smegmatis, M. triplex, M. tuberculosis, M. ulcerans, and M. xenopi.
According to the second exemplary embodiment, the present disclosure provides an antibacterial composition having antibacterial activity against bacteria belonging to the genus Mycobacterium, which includes a Bacillus velezensis PMC 206 strain or a culture thereof.
According to the present disclosure, the Bacillus velezensis PMC 206 strain may be extracted from vinegar. For example, Bacillus velezensis PMC 206 strain is assigned Accession No. KCTC 15187BP.
According to the present disclosure, the bacteria belonging to the genus Mycobacterium may be one or more selected from the group consisting of M. abscessus, M. africanum, M. asiaticum, M. avium complex (MAC), M. bovis, M. chelonae, M. fortuitum, M. gordonae, M. haemophilum, M. intracellulare, M. kansasii, M. lentiflavum, M. leprae, M. malmoense, M. marinum, M. microti, M. phlei, M. scrofulaceum, M. smegmatis, M. triplex, M. tuberculosis, M. ulcerans, and M. xenopi.
In the present disclosure, the antibacterial composition may be used in living organisms.
In the present disclosure, the antibacterial composition may be used in an in vitro environment.
According to the third exemplary embodiment, the present disclosure provides a pharmaceutical composition for treating or preventing tuberculosis having antibacterial activity against bacteria belonging to the genus Mycobacterium, which includes a Bacillus velezensis PMC 206 strain or a culture thereof as an active ingredient.
According to the present disclosure, the Bacillus velezensis PMC 206 strain may be extracted from vinegar. For example, Bacillus velezensis PMC 206 strain is assigned Accession No. KCTC 15187BP.
According to the present disclosure, the bacteria belonging to the genus Mycobacterium may be one or more selected from the group consisting of M. abscessus, M. africanum, M. asiaticum, M. avium complex (MAC), M. bovis, M. chelonae, M. fortuitum, M. gordonae, M. haemophilum, M. intracellulare, M. kansasii, M. lentiflavum, M. leprae, M. malmoense, M. marinum, M. microti, M. phlei, M. scrofulaceum, M. smegmatis, M. triplex, M. tuberculosis, M. ulcerans, and M. xenopi.
In the present disclosure, the tuberculosis may include one or more selected from the group consisting of ocular tuberculosis, skin tuberculosis, adrenal tuberculosis, renal tuberculosis, epididymal tuberculosis, lymphatic tuberculosis, laryngeal tuberculosis, middle ear tuberculosis, intestinal tuberculosis, multidrug-resistant tuberculosis, pulmonary tuberculosis, biliary tuberculosis, bone tuberculosis, throat tuberculosis, breast tuberculosis, and spinal tuberculosis.
In the present disclosure, the tuberculosis may be latent tuberculosis.
In the present disclosure, the pharmaceutical composition may further include at least one selected from the group consisting of rifampicin, rifapentine, isoniazid, pyrazinamide, ethambutol, streptomycin, fluoroquinolone, kanamycin, cycloserine, prothionamide, levofloxacin, moxifloxacin, ofloxacin, rifabutin, capeomycin, amikacin, ciprofloxacin, protionamide, ethionamide, cycloserine, thioacetazone, clofazimine, amoxicillin/clavulanate, a derivative of dianomidiphenylsulphone, clarithromycin, azithromycin, and linezolid.
According to the fourth exemplary embodiment, the present disclosure provides a food composition for preventing or improving tuberculosis having antibacterial activity against bacteria belonging to the genus Mycobacterium, which includes the Bacillus velezensis PMC 206 strain or a culture thereof as an active ingredient.
According to the present disclosure, the Bacillus velezensis PMC 206 strain may be extracted from vinegar. For example, Bacillus velezensis PMC 206 strain is assigned Accession No. KCTC 15187BP.
According to the present disclosure, the bacteria belonging to the genus Mycobacterium may be one or more selected from the group consisting of M. abscessus, M. africanum, M. asiaticum, M. avium complex (MAC), M. bovis, M. chelonae, M. fortuitum, M. gordonae, M. haemophiluscellulare, M. kansasii, M. lentiflavum, M. leprae, M. malmoense, M. marinum, M. microti, M. phlei, M. scrofulaceum, M. smegmatis, M. triplex, M. tuberculosis, M. ulcerans, and M. xenopi.
In the present disclosure, the tuberculosis may include one or more selected from the group consisting of ocular tuberculosis, skin tuberculosis, adrenal tuberculosis, renal tuberculosis, epididymal tuberculosis, lymphatic tuberculosis, laryngeal tuberculosis, middle ear tuberculosis, intestinal tuberculosis, multidrug-resistant tuberculosis, pulmonary tuberculosis, biliary tuberculosis, bone tuberculosis, throat tuberculosis, breast tuberculosis, and spinal tuberculosis.
In the present disclosure, the tuberculosis may be extensive drug-resistance (XRD) tuberculosis.
The present disclosure provides a Bacillus velezensis PMC 206 strain which has antibacterial activity. More specifically, the strain has antibacterial activity against Mycobacterium tuberculosis. Therefore, the strain may be used for treating, preventing, or improving tuberculosis.
In particular, the strain has the advantage in that it has no side effects due to drug resistance and long-term high-dose drug treatment, compared to compound-based drug treatment.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which the present disclosure pertains. All publications, patents, and other references mentioned herein are incorporated by reference in their entirety.
As used herein, the term “extensive drug-resistance (XRD)” refers to drug resistance, and particularly, resistance to a wide range of drugs.
As used herein, the term “antibacterial” refers to killing microorganisms or inhibiting the growth of microorganisms. In one embodiment, as used herein, the term may be used to mean killing a specific microorganism or inhibiting the growth of a specific microorganism. In a specific embodiment, as used herein, the term may be used to mean killing bacteria belonging to the genus Mycobacterium or inhibiting its growth. In a more specific embodiment, as used herein, the term may be used to mean killing one or more bacteria selected from the group consisting of M. abscessus, M. africanum, M. asiaticum, M. avium complex (MAC), M. bovis, M. chelonae, M. fortuitum, M. gordonae, M. haemophilum, M. intracellulare, M. kansasii, M. lentiflavum, M. leprae, M. malmoense, M. marinum, M. microti, M. phlei, M. scrofulaceum, M. smegmatis, M. triplex, M. tuberculosis, M. ulcerans, and M. xenopi, or inhibiting their growth.
As used herein, the term “prevention” refers to any action without limitation that blocks the symptoms caused by tuberculosis or inhibits or delays the onset of the symptoms by using the composition of the present disclosure, which includes a Bacillus velezensis PMC 206 strain as an active ingredient.
As used herein, the terms “improvement” and “treatment” refer to any action without limitation that improves the symptoms caused by tuberculosis or becomes beneficial, by using the composition of the present disclosure, which includes a Bacillus velezensis PMC 206 strain as an active ingredient.
Hereinafter, the specific details of the present disclosure are described.
Strains Obtained from Fermented Foods
The present disclosure intends to provide a Bacillus velezensis PMC 206 strain. The Bacillus velezensis PMC 206 strain may be obtained from fermented foods. In an exemplary embodiment, the Bacillus velezensis PMC 206 strain may be extracted from vinegar. For example, the Bacillus velezensis PMC 206 strain is assigned Accession No. KCTC 15187BP. 16S rRNA sequence of KCTC15187BP is SEQ ID NO: 1.
The present disclosure intends to provide an antibacterial composition including a Bacillus velezensis PMC 206 strain. Since the strain is non-cytotoxic, it may be included in compositions which are used in vivo and in vitro.
The antibacterial composition may be an antibacterial composition administered to a living body. In particular, the living organism refers to a living individual, and the individual includes mammals including humans. That is, the present disclosure provides, in an embodiment, an antibacterial composition which has antibacterial activity against undesired microorganisms present in mammals including humans. In a specific embodiment, the present disclosure provides an antibacterial composition having antibacterial activity against pathogenic bacteria present in mammals including humans.
The antibacterial composition including the Bacillus velezensis PMC 206 strain may be an antibacterial composition that has antibacterial activity against undesired microorganisms existing in an in vitro environment. In a specific embodiment, the antibacterial composition including the Bacillus velezensis PMC 206 strain provides an antibacterial composition having antibacterial activity against pathogenic bacteria present in an in vitro environment. In particular, the term “in vitro environment” is an exemplary embodiment and may refer to the surface of a material that needs to be washed. In particular, the term “washing” may refer to removal of undesired microorganisms. In particular, the term “removal” may refer to killing some bacteria, inhibiting the growth of bacteria, or reducing the number of bacteria.
The surface of the material may include the surface of household goods including furniture, dishware, home appliances, etc., but is not limited thereto and includes all surfaces of materials that need to be washed. That is, the present disclosure provides an antibacterial composition having antibacterial activity against pathogenic microorganisms existing in an in vitro environment.
The present disclosure also intends to provide an antibacterial method using an antibacterial composition including a Bacillus velezensis PMC 206 strain.
The antibacterial method may include administering the antibacterial composition to a living body. The living organism refers to a living individual, and the individual includes mammals including humans. That is, the present disclosure provides an antibacterial method which includes administering the composition to mammals including humans.
The antibacterial method may also include treating the antibacterial composition in vitro. The in vitro refers to the surface of a material that needs to be washed, and the present disclosure provides an antibacterial method for treating the surface of a material that needs to be washed with the antibacterial composition.
The present disclosure provides a Bacillus velezensis PMC 206 strain, an antibacterial composition including the strain, and a pharmaceutical composition and a food composition for treating, preventing, and improving tuberculosis and inflammatory diseases including the strain. In particular, in one embodiment, the composition may include the Bacillus velezensis PMC 206 strain in the form of live cells, dead cells, or a culture thereof. As used herein, the culture may be a culture including the Bacillus velezensis PMC 206 strain, or it may include its lysate, a supernatant or pellet after centrifugation, a concentrate, a dried material, etc.
The present disclosure provides an antibacterial composition including a Bacillus velezensis PMC 206 strain. The antibacterial composition may have antibacterial activity against bacteria belonging to the genus Mycobacterium. In a specific embodiment, the antibacterial composition may have antibacterial activity against one or more bacteria selected from the group consisting of M. abscessus, M. africanum, M. asiaticum, M. avium complex (MAC), M. bovis, M. chelonae, M. fortuitum, M. gordonae, M. haemophilum, M. intracellulare, M. kansasii, M. lentiflavum, M. leprae, M. malmoense, M. marinum, M. microti, M. phlei, M. scrofulaceum, M. smegmatis, M. triplex, M. tuberculosis, M. ulcerans, and M. xenopi. In a more specific embodiment, the antibacterial composition may have antibacterial activity against M. tuberculosis.
The present disclosure provides a pharmaceutical composition for treating or preventing tuberculosis including a Bacillus velezensis PMC 206 strain.
As used herein, the term “tuberculosis” refers to an infectious disease transmitted by Mycobacterium tuberculosis. For example, the tuberculosis may be one or more selected from the group consisting of ocular tuberculosis, skin tuberculosis, adrenal tuberculosis, renal tuberculosis, epididymal tuberculosis, lymphatic tuberculosis, laryngeal tuberculosis, middle ear tuberculosis, intestinal tuberculosis, multidrug-resistant tuberculosis, pulmonary tuberculosis, biliary tuberculosis, bone tuberculosis, throat tuberculosis, lung deficiency syndrome, breast tuberculosis, and spinal tuberculosis, but is not limited thereto.
In a pharmaceutical composition for treating or preventing tuberculosis according to the present disclosure, the pharmaceutical composition may further include a carrier. For example, the pharmaceutically acceptable carrier may be one which is commonly used in preparation, and may include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc., but is not limited thereto. The pharmaceutical composition may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, etc., in addition to the ingredients described above. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).
In the pharmaceutical composition for treating or preventing tuberculosis according to the present disclosure, the pharmaceutical composition may be prepared by mixing one or more diluents or excipients such as such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants commonly used in this technical field. For example, solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, etc. These solid preparations are prepared by mixing the strain of the present disclosure or a culture thereof with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. Additionally, in addition to simple excipients, lubricants such as magnesium styrate talc may also be used. Liquid preparations for oral administration may include suspensions, oral solutions, emulsions, syrups, etc., for example, may include wetting agents, sweeteners, fragrances, preservatives, etc. may be included. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories, etc. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil (e.g., olive oil), and injectable ester (e.g., ethyl oleate), etc. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurinum, glycerol, gelatin, etc. may be used.
In the pharmaceutical composition for treating or preventing tuberculosis according to the present disclosure, the pharmaceutical composition may further include materials that can improve the effects of preventing or treating diseases. For example, the pharmaceutical composition may further include at least one selected from the group consisting of rifampicin, rifapentine, isoniazid, pyrazinamide, ethambutol, streptomycin, fluoroquinolone, kanamycin, cycloserine, prothionamide, levofloxacin, moxifloxacin, ofloxacin, rifabutin, capeomycin, amikacin, ciprofloxacin, protionamide, ethionamide, thioacetazone, clofazimine, amoxicillin/clavulanate, a derivative of dianomidiphenylsulphone, clarithromycin, azithromycin, and linezolid. According to an exemplary embodiment, the pharmaceutical composition may further include rifampicin in the Bacillus velezensis PMC 206 strain.
The present disclosure provides a method for treating tuberculosis which includes administering, to an individual, a composition including a Bacillus velezensis PMC 206 strain. In particular, the individual may refer to a mammal including humans.
In the method for treating tuberculosis according to the present disclosure, the composition may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method, and the dose varies depending on the patient's condition and weight, degree of disease, drug type, and route and time of administration, but may be appropriately selected by those skilled in the art.
In the method for treating tuberculosis according to the present disclosure, the composition is administered in a pharmaceutically effective amount. The “pharmaceutically effective amount” refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level may be determined depending on factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the field of medicine. The composition of the present disclosure may be administered as an individual treatment or in combination with other treatments, may be administered sequentially or simultaneously with conventional treatments, and may be administered once or multiple times. Considering all of the factors described above, it is important to administer an amount that may achieve a maximum effect with a minimum amount without side effects, and this may easily be determined by those skilled in the art. In particular, the composition of the present disclosure and rifampicin may be administered together. When rifampicin is administered together, the effects of prevention and treatment of the disease caused by the composition of the present disclosure may be improved. Specifically, the effective amount of the composition according to the present disclosure may vary depending on the patient's age, sex, and body weight, generally 0.1 mg to 100 mg, preferably 0.5 mg to 10 mg per kg of body weight, and may be administered daily or every other day, or may be administered in 1 to 3 divided doses daily. However, since the dose may increase or decrease depending on the route of administration, severity of obesity, sex, body weight, age, etc., the dose does not limit the scope of the present disclosure in any way.
The present disclosure provides a food composition for preventing or improving tuberculosis including a Bacillus velezensis PMC 206 strain. In particular, the food composition includes a health functional food composition, and although the health functional food is not particularly limited as long as it includes the Bacillus velezensis PMC 206 strain, the strain may be included in an amount of 0.01 wt % to 20 wt % based on the total weight of the health functional food composition.
In the food composition for preventing or improving tuberculosis according to the present disclosure, the food composition is not particularly limited as long as it is a food that is commonly prepared and/or sold. For example, the food composition includes meats, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice creams, various kinds of soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, which may be used in the form of pills, powders, granules, precipitates, tablets, capsules, or beverages, and the food composition includes all health functional foods in the conventional sense.
In the food composition for preventing or improving tuberculosis according to the present disclosure, the food composition has no particular limitation on liquid ingredients other than including the strain described above, it may include various kinds of flavoring agents or natural carbohydrates (e.g., as common beverages) as additional ingredients.
Examples of carbohydrates include conventional sugars such as monosaccharides (e.g., glucose, fructose, etc.); disaccharides (e.g., maltose, sucrose, etc.); polysaccharides (e.g., dextrin, cyclodextrin); etc. and sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). As flavoring agents other than those described above, natural flavoring agents (thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be advantageously used.
In the food composition for preventing or improving tuberculosis according to the present disclosure, the food composition may include, in addition to the additional ingredients described above, various kinds of nutrients, vitamins, minerals (electrolytes), flavoring agents (e.g. synthetic and natural flavoring agents), coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated drinks, etc.
Hereinafter, the present disclosure will be described in detail through examples. In particular, these examples are solely for describing the present disclosure more specifically, and it is obvious to those skilled in the art that the scope of the present disclosure is not limited by these examples.
1. Isolation, Identification, and Genetic Characterization of Probiotic Strain from Vinegar
A probiotic strain was isolated from Korean traditional fermented vinegar and cultured in a food grade medium. The probiotic strain was cultured in a food-grade medium (glucose 20 g/L, yeast peptone standard-type F 25 g/L, Tween 80 1 g/L, magnesium sulfate 0.2 g/L, pH 6.3). The isolation, cultivation, and storage of the strain were performed as described in a previous report [Yoon Y, Seo H, Kim S, et al. Anti-Tuberculosis Activity of Pediococcus acidilactici Isolated from Young Radish Kimchi against Mycobacterium tuberculosis. J Microbiol Biotechnol. 2021 Dec. 28; 31 (12): 1632-1642.]. The strain was confirmed by sequence analysis of 16s rRNA gene.
The sequencing of 16s rRNA gene was performed by Biofact (Korea) and was compared with the GenBank database using the basic local alignment search tool (BLAST) in the National Center for Biotechnology Information (NCBI). For the sequencing of the whole genome the strain, gDNA was extracted from the strain, sequenced using PacBio RSII (PacBio, USA) at CJ Bioscience (Korea), and analyzed with the Ezbiocloud server. The information of the genome of the Bacillus beregensis PMC206 strain was deposited on the NCBI website under Accession No. PRJNA865468.
Murine Raw 264.7 macrophages which have reached confluence were infected with M. tuberculosis, treated with probiotics or drugs according to the present disclosure, cultured for 3 days, and then CFU analysis was performed. H37Rv and XDR of the Mycobacterium tuberculosis strain used in this study were purchased from American Type Culture Collection (ATCC, USA) and Korean Mycobacterium Resource Center (KMRC, Korea), respectively. The synergistic effect of rifampicin (RIF) and isoniazid (INH), which are representative first-line drugs for the treatment of tuberculosis, was also examined.
The in vivo anti-tuberculosis effect of probiotics according to the present disclosure was evaluated in a mouse model of XDR pulmonary tuberculosis and a mouse model of latent tuberculosis. First, 6-week-old Balb/c mice were infected with the XDR strain by high-dose lung inhalation using a nebulizer, and were then administered with the probiotic strain by lung inhalation three times a day starting one week after infection. In a mouse model of latent pulmonary tuberculosis, mice were infected with a low-dose of the XDR strain, and one month thereafter, the mice were administered with the probiotic strain by lung inhalation once a day, twice a week for 8 months. Rifampicin was used as a control drug and was administered orally at 30 mg/kg. In this animal experiment, saline was used as a vehicle. For further analysis, lungs were extracted and M. tuberculosis was quantitatively analyzed by CFU and real-time PCR. Primers used were KY18 (forward, 5′-CACATGCAAGTCGAACGGAAAGG-3′; reverse, 5′-CCTTTCCGTTCGACTTGCATGTG-3′) and KY75 (forward, 5′-GCCCGTATCGCCCGCACGCTCACA-3′; reverse, 5′-TGTGAGCGTGCGGGCGATACGGGC-3′). Perfused lungs were used to examine the lung microenvironment in the mouse model of XDR pulmonary tuberculosis. Microbiome and metabolite analyses were performed according to previously reported methods. Lung cytokines were analyzed using a mouse ELISA kit (Thermo Fisher Scientific, USA), and the hematoxylin and eosin (H&E) staining analysis was performed in the Korea Pathology Technical Center (KPNT, Korea).
A probiotic strain isolated from vinegar, a traditional fermented food, was identified based on the gene sequence of 16s rRNA and then confirmed by the analysis of the whole genome. As a result, it was found that the genome of the isolated probiotic strain consisted of a 3,990,015 bp single circular chromosome with 3,841 coding sequences (CDS) (
2. Evaluation of Antimycobacterial Activity of Bacillus velezensis PMC 206
The antimycobacterial activity of the Bacillus velezensis PMC 206 strain according to the present disclosure was confirmed using an in vitro macrophage infection model. Raw 264.7 cells were infected with the H37Rv strain, treated with the PMC 206 extract for 3 days, and then quantified through colony-forming unit (CFU) analysis. As a result, it was found that the PMC206 extract significantly inhibited both H37Rv (
3. Evaluation of Efficacy of Bacillus velezensis PMC 206 in a Mouse Model of Fatal XDR Pulmonary Tuberculosis
The in vivo efficacy of the Bacillus velezensis PMC 206 strain according to the present disclosure was evaluated using a mouse model of XDR pulmonary tuberculosis. Six-week-old Balb/c mice were infected with the XDR strain by high-dose lung inhalation using a nebulizer, and were then administered in the same manner with the Bacillus velezensis PMC 206 strain once a day and three times a week, and the mortality was observed. As a result, the infected mice died on the 12th week after the end of the experiment (
4. Evaluation of Efficacy of Bacillus velezensis PMC 206 in Mouse Model of Latent Tuberculosis
As a result of evaluating the efficacy of the Bacillus velezensis PMC 206 strain in a model of latent tuberculosis induced by low-dose infection with the XDR strain, no significant change in body weight was observed upon administration of PMC206 (
5. Analysis of Characteristics of Intrapulmonary Microenvironment in Treating M. tuberculosis by Bacillus velezensis PMC 206
In order to examine the characteristics of the intrapulmonary microenvironment in treating M. tuberculosis by the Bacillus velezensis PMC 206 strain according to the present disclosure, granuloma formation and immune response analysis were performed. As a result, unlike in uninfected mice, granulomas were formed in the lungs of mice infected with M. tuberculosis in both models, and while relatively few granulomas were formed in PMC206-treated mice (
As a result of performing the analysis of 16S rRNA gene-based lung metagenomics in a mouse model of lethal pulmonary tuberculosis, while the occupancy of M. tuberculosis in the lungs of mice treated with PMC206 decreased compared to that of the infected control group, it did not decrease in the groups treated with INH or RIF (
6. Effect of Autophagy Induction by Bacillus velezensis PMC 206
As a result of confirming the profiling regarding the induction of autophagy by PMC206 in a macrophage model infected with M. tuberculosis, PMC206 was shown to increase autophagic vacuoles in the model (
7. Synergistic and Preventive Effects of Bacillus velezensis PMC 206 on Pulmonary Tuberculosis
As a result of confirming additional synergistic and preventive effects in a macrophage model infected with M. tuberculosis, it was confirmed that while RIF at 1 mg/kg/day did not significantly reduce the burden of M. tuberculosis in the lungs throughout the entire experiment, a significant reduction could be achieved when the same concentration of RIF and PMC206 were administered in combination (
As described above, specific parts of the present disclosure have been described in detail; however, it is obvious to those skilled in the art that these specific techniques are merely preferred exemplary embodiments and the scope of the present disclosure is not limited thereto. Accordingly, the actual scope of the present disclosure will be defined by the accompanying claims and equivalents thereof.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2023-0142894 | Oct 2023 | KR | national |
