KR102547036B1 - Composition for preventing or treating degenerative brain disease containing Blautia obeum as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or treating degenerative brain diseases. According to the composition containing Blautia obeum, endoplasmic reticulum derived therefrom, lysate, culture medium or a mixture thereof as an active ingredient, the composition has an effect of being able to be usefully used for preventing, alleviating or treating degenerative brain diseases.

Description

Composition for preventing or treating degenerative brain disease containing Blautia obeum as an active ingredient}

It relates to a composition for preventing or treating degenerative brain disease.

Degenerative brain disease is a disease in which degenerative changes appear in nerve cells of the central nervous system and cause various symptoms such as impairment of motor and sensory functions, inhibition of higher-order causative functions such as memory, learning, and computational reasoning. Types of degenerative brain diseases include Alzheimer's disease, Parkinson's disease, and memory disorders. In degenerative brain diseases, the death of nerve cells occurs by rapid or slow necrosis or apoptosis.

The reason why studies on degenerative brain diseases are continuously conducted is that the number of patients with dementia is rapidly increasing and the social cost of treating and managing them is very high. Treatments are being developed in many countries to alleviate the incidence itself, but it was confirmed that drugs already on the market temporarily suppress the manifestation of symptoms rather than improving some of the symptoms of the disease, which reduces the efficacy of the drug. As a result of having a larger range of change, the disadvantage of overdose of the drug occurs, and finally, the expression of side effects caused by the drug is higher, causing difficulties for both the patient and the guardian. Recently, natural treatments using lactic acid bacteria are being developed to increase the safety and effectiveness of drugs. Because. In other words, medicines applied to neurological diseases and mental stress, including degenerative brain diseases, have been found to be effective in improving symptoms, but general medicines are composed of chemical components, so long-term intake Possibility of deterioration in physical parts such as reduced exercise, reduced efficacy, and reduced liver function is very high, so it is highly likely to reduce the probability of side effects that occur when using lactic acid bacteria as a substitute for natural medicines.

Accordingly, the present inventors have completed the present invention by confirming that Blautia obeum is effective for degenerative brain diseases.

One aspect is to provide a pharmaceutical composition for preventing or treating degenerative brain disease comprising Blautia obeum , endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof as an active ingredient will be.

Another aspect provides a health functional food composition for preventing or improving degenerative brain disease comprising Blautia obeum, endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof as an active ingredient. is to do

Another aspect is to provide a feed composition for preventing or improving degenerative brain disease comprising Blautia obeum, endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof as an active ingredient will be.

One aspect provides a pharmaceutical composition for preventing or treating degenerative brain disease comprising Blautia obeum , endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof as an active ingredient. .

The Blautia obeum may be deposited under accession number KCTC 14560BP.

The Blautia obeum may be live or dead cells.

As used herein, the term "vesicle" refers to particles secreted from cells and released into the extracellular space, including exosomes, ectosomes, microvesicles, and microparticles. , exosome like vesicles, and the like. Extracellular endoplasmic reticulum can reflect the state of the secreting cell of origin (donor cell), show various biological activities depending on which cell it is secreted from, and play an important role in cell-to-cell interactions by transferring genetic material and proteins between cells. can do. In addition, cell-derived substances including the endoplasmic reticulum cause disease or stimulate immune cells to fight against disease, and have the effect of helping to decompose and absorb substances that humans cannot digest through the metabolic process of microorganisms. there is. The endoplasmic reticulum is a membrane-structured endoplasmic reticulum, and the inside and the outside are divided, and has a plasma membrane lipid, a plasma membrane protein, a nucleic acid, and a cytoplasmic component of the cell, and is larger than the original cell. may be small.

In one embodiment, the endoplasmic reticulum may be separated from a cell lysate of a culture medium of a Blautia obeum strain.

In one embodiment, the extracellular vesicles may have a diameter of 10 nm to 400 nm. For example, it may be 10 nm to 400 nm, 10 nm to 350 nm, 10 nm to 300 nm, or 10 nm to 250 nm.

As used herein, the term "culture medium" may be used interchangeably with "culture supernatant", "conditioned culture medium" or "conditioned medium", and may supply nutrients so that Blautia obeum can grow and survive in vitro. It may mean the entire medium including the strain obtained by culturing the strain in a medium for a certain period of time, its metabolites, extra nutrients, and the like. In addition, the culture solution may mean a culture solution obtained by removing the cells from the cell culture solution obtained by culturing the strain. On the other hand, the liquid from which the cells are removed from the culture solution is also called "supernatant". It can be obtained by removing the precipitate and taking only the upper liquid. The "cell" refers to the "strain" itself of the present invention, and includes the "strain" itself separated and selected from skin samples, etc., or the "strain" separated from the culture solution by culturing the "strain". The cells can be obtained by centrifuging the culture solution and taking the part that has sunk in the lower layer, or can be obtained by leaving it for a certain period of time and then removing the upper liquid as it sinks to the lower layer of the culture medium by gravity.

The culture solution may include a culture solution itself obtained by culturing the strain, a concentrate thereof, or a lyophilized product or a culture supernatant obtained by removing the strain from the culture solution, a concentrate thereof, or a lyophilisate.

The culture medium is obtained by culturing Blautia obeum in an appropriate medium (eg, R2A medium or TSA medium) at any temperature above 10 ° C. or below 40 ° C. for a certain period of time, for example, 4 to 50 hours. it could be

In one embodiment, the culture supernatant of the strain may be obtained by centrifuging or filtering the strain culture medium to remove the strain.

In another embodiment, the concentrate may be obtained by concentrating the supernatant obtained after filtering the strain culture medium itself, or the culture medium using a centrifugal separation or filter.

The culture medium and culture conditions for culturing the Blautia obeum can be appropriately selected or modified by those skilled in the art.

In this specification, the term "lysate" may mean a product obtained by disrupting the cell wall of the strain itself by chemical or physical force.

As used herein, the term "culture broth extract" refers to an extract obtained from the culture medium or a concentrate thereof, and may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof, or a fraction obtained by fractionating the same. can

The degenerative brain diseases include dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, cognitive impairment, learning disorder, cerebral amyloid angiopathy, Down's syndrome, amyloidogenic stroke, systemic Amyloid disease, Dutch type amyloidosis, Niemann-Pick disease, senile dementia, amyotrophic lateral sclerosis, ataxia, spinocerebellar atrophy, Tourette's syndrome ), Friedrich`s Ataxia, Machado-Joseph`s disease, Lewy Body Dementia, Dystonia, Progressive Supranuclear Palsy and It may be selected from the group consisting of Frontotemporal Dementia.

As used herein, the term "treat" may mean that degenerative brain diseases are cured in a shorter time compared to natural healing. The treatment may include prevention, improvement and/or alleviation of degenerative brain diseases. In addition, the treatment may include all treatments of diseases related to degenerative brain diseases.

The term "prevention" is used in the sense of including all mechanisms and/or effects that act on a subject not having a specific disease to prevent the development of the specific disease, to delay the onset of the disease, or to reduce the incidence of the disease.

As used herein, the term "pharmaceutically effective amount" or "active ingredient" refers to any composition used in the practice of the invention provided herein that is sufficient to alleviate, inhibit the progression of, or prevent a disease, disorder or condition, or one or more symptoms thereof. can mean the amount of

As used herein, the terms "administering," "applying," "introducing," and "implanting" are used interchangeably and a method or route that results in at least partial localization of a composition to a desired site according to one embodiment. By can mean the placement of a composition according to one embodiment into an individual.

The term "pharmaceutical composition" can refer to a molecule or compound that imparts some beneficial effect upon administration to a subject. Beneficial effects may include enabling diagnostic determination; amelioration of a disease, symptom, disorder or condition; reducing or preventing the occurrence of a disease, condition, disorder or condition; and responding to a disease, symptom, disorder or condition in general.

The pharmaceutical composition, in addition to the active ingredient, pharmaceutically acceptable carriers, excipients, diluents, fillers, bulking agents, wetting agents, disintegrants, emulsifiers (surfactants), lubricants, sweeteners, flavoring agents, suspending agents, preservatives, etc. It may further include one or more adjuvants selected from the group consisting of. The adjuvant may be appropriately adjusted according to the formulation to which the pharmaceutical composition is applied, and one or more adjuvants may be selected from among all adjuvants commonly used in the pharmaceutical field. In one embodiment, the pharmaceutically acceptable carrier is one commonly used in the formulation of drugs, saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes and these components One or more of the components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate formulations for injections such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets, and may act specifically on target organs. A target organ-specific antibody or other ligand may be used in combination with the carrier. Furthermore, it can be preferably formulated according to each disease or component by using an appropriate method in the art or by using a method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA). there is.

An effective amount of the active ingredient or the pharmaceutical composition can be administered orally or parenterally during clinical administration and can be used in the form of a general pharmaceutical preparation. Parenteral administration may refer to administration through an administration route other than oral, such as rectal, intravenous, peritoneal, intramuscular, arterial, transdermal, nasal, inhalation, ocular or subcutaneous administration, and may be administered by local administration to the lesion site. can Upon oral administration, the pharmaceutical composition may be formulated into a formulation capable of coating the active ingredient or protecting it from decomposition in the stomach in order to prevent the active ingredient from being decomposed in the stomach. In one embodiment, the composition is administered by an administration route selected from the group consisting of oral administration, intravenous administration, intraperitoneal administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration and intrarectal administration. may be administered. When the pharmaceutical composition of the present invention is used as a medicine, it may additionally contain one or more active ingredients exhibiting the same or similar functions.

The pharmaceutical composition may be formulated in the form of a solution, suspension, syrup or emulsion in an aqueous or oily medium, or in the form of a powder, powder, granule, tablet or capsule, and additionally contains a dispersing agent or stabilizer for formulation. can do. When formulating the pharmaceutical composition, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are commonly used. Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, Witepsol, Macrogol, Tween 61, cacao butter, liurine fat, glycerogeratin and the like may be used.

The pharmaceutical composition may be mixed with various pharmaceutically acceptable carriers such as physiological saline or organic solvents, and carbohydrates such as glucose, sucrose or dextran, ascorbic acid (Ascorbic acid) may be used to increase stability or absorption. Acid) or glutathione (Antioxidants), chelating agents (Chelating agents), low molecular weight protein or other stabilizers (Stabilizers) can be used as a drug.

The pharmaceutical composition may be administered in a pharmaceutically effective amount. The administration dose is not particularly limited, and may vary depending on absorption in the body, body weight, patient's age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease. The pharmaceutical composition of the present invention is prepared in consideration of the effective amount range, and the unit dosage formulation formulated in this way can be administered according to the judgment of an expert who monitors or observes the administration of drugs as necessary and a specialized dosing method according to individual needs. It can be used or administered several times at regular time intervals. The dosage of the pharmaceutical composition may be 1 ug/kg/day to 1,000 mg/kg/day, but is not limited thereto. The daily or one-time dosage may be formulated as one preparation in unit dosage form, formulated in appropriate portions, or prepared by placing it in a multi-dose container.

The dosage of Blautia obeum may be an amount sufficient to induce an effective response to degenerative brain disease in a patient. In certain embodiments of the present specification, the Blautia obeum may be 10 ³ to 10 ¹¹ , 10 ⁴ to 10 ¹⁰ , 10 ⁶ to 10 ¹⁰ , or 10 ⁶ to 10 ⁹ . Alternatively, the dose may be 0.01 mg to 10 mg or 0.1 mg to 1 mg of Blautia obeum provided as a suspension or dry formulation. In another example, the dose is 1 X 10 ³ to 1 X 10 ¹¹ cfu/ml, 1 X 10 ⁴ to 1 X 10 ¹⁰ cfu/ml, 1 X 10 ⁶ to 1 X 10 ¹⁰ cfu/ml, or 1 X 10 ⁶ to 1 X 10 ⁹ cfu/ml.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be an individual in need of healing of a degenerative brain disease.

Another aspect is to treat degenerative brain disease, including the step of administering Blautia obeum, endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof to a subject suffering from a degenerative brain disease. provides a way to

Another aspect provides a method for preventing degenerative brain disease, comprising administering Blautia obeum, endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof to a subject. do.

The method, prior to the administering step, may further include identifying a subject in need of prevention and/or treatment of a degenerative brain disease.

Another aspect is a functional food composition for preventing or improving degenerative brain disease comprising Blautia obeum , endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof as an active ingredient. to provide.

The strain, endoplasmic reticulum, lysate, culture medium and their effects are as described above.

The health functional food is a food manufactured using nutrients that are easily deficient in daily meals or raw materials or ingredients having functions useful to the human body (hereinafter referred to as 'functional raw materials'), to maintain health, prevent certain diseases or symptoms, and / or means any food that helps to improve, and there is no particular restriction on the form of the final product. For example, the health functional food may be selected from the group consisting of various foods, beverage compositions, food additives, etc., but is not limited thereto.

The content of the active ingredient contained in the health functional food is appropriately not particularly limited depending on the type of food, desired use, etc., for example, 0.0001 to 99% by weight, 0.0001 to 95% by weight, 0.0001 to 90% by weight of the total food weight. %, 0.0001 to 80% by weight, 0.0001 to 50% by weight, 0.001 to 99% by weight, 0.001 to 95% by weight, 0.001 to 90% by weight, 0.001 to 80% by weight, 0.001 to 50% by weight, 0.01 to 99% by weight, 0.01 to 95%, 0.01 to 90%, 0.01 to 80%, 0.01 to 50%, 0.1 to 99%, 0.1 to 95%, 0.1 to 90%, 0.1 to 80%, 0.1 to 99% 50 wt%, 0.1 to 30 wt%, 0.1 to 10 wt%, 1 to 99 wt%, 1 to 95 wt%, 1 to 90 wt%, 1 to 80 wt%, 1 to 50 wt%, 1 to 30 wt% %, 1 to 10% by weight, 10 to 99% by weight, 10 to 95% by weight, 10 to 90% by weight, 10 to 80% by weight, 10 to 50% by weight, 10 to 30% by weight, 25 to 99% by weight, 25 to 95% by weight, 25 to 90% by weight, 25 to 80% by weight, 25 to 50% by weight, 25 to 30% by weight, 40 to 99% by weight, 40 to 95% by weight, 40 to 90% by weight, 40 to 80 wt%, 40 to 50 wt%, 50 to 99 wt%, 50 to 95 wt%, 50 to 90 wt%, 50 to 80 wt%, 60 to 99 wt%, 60 to 95 wt%, 60 to 90 wt% %, or 60 to 80% by weight, but is not limited thereto.

The health functional food includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors or natural flavors, colorants, enhancers (cheese, chocolate, etc.), pectic acid or its salts, alginic acid or its salts, It may further contain at least one selected from the group consisting of organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. The ratio of these additives is generally selected from the range of 0.001 to about 20 parts by weight per 100 parts by weight of the total health functional food, but is not limited thereto.

Another aspect provides a feed composition for preventing or improving degenerative brain disease comprising Blautia obeum, endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a mixture thereof as an active ingredient. .

The strain, endoplasmic reticulum, lysate, culture medium and their effects are as described above.

When used as a feed composition, the composition may be 20 to 90% high concentration or prepared in powder or granular form. The feed composition includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid and malic acid, sodium phosphate, potassium phosphate, acidic pyrophosphate, polyphosphate (polyphosphate), polyphenol, catechin, alpha-tocopherol, and rosemary. extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid, and the like, or any one or more natural antioxidants may be further included. The composition may be formulated in the form of a common feed, and may include common feed ingredients together.

The feed includes grains such as milled or crushed wheat, oats, barley, corn and rice; vegetable protein feeds such as those based on rape, soybean, and sunflower; animal protein feed such as blood meal, meat meal, bone meal and fish meal; It may further include sugar and dairy products, for example, dry ingredients composed of various powdered milk and whey powder, etc., and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.

The feed composition may be administered to animals alone or in combination with other feed additives in an edible carrier. In addition, the feed additives can be easily administered to animals as a top dressing, directly mixed with animal feed, or in an oral formulation separate from feed. When the feed additive is administered separately from animal feed, as is well known in the art, it can be prepared as an immediate release or sustained release formulation by combining it with an edible carrier acceptable for food science. Such edible carriers can be solid or liquid, for example corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the feed additive may be a tablet, capsule, powder, troche or sugar-containing tablet or top dressing in a microdispersible form. When a liquid carrier is used, the feed additive may be a gelatin soft capsule, or a syrup, suspension, emulsion, or solution formulation.

In addition, the feed may contain auxiliary agents, such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like. The feed additive may be used by adding it to the animal's feed by soaking, spraying or mixing.

The feed or feed additive of the present invention can be applied to a number of animal diets including mammals, poultry and fish.

As the mammal, it can be used for pigs, cows, sheep, goats, laboratory rodents, and laboratory rodents, as well as pets (eg dogs, cats), etc., and as the poultry, chickens, turkeys, ducks, geese, pheasants, And it can also be used for quail, etc., and can be used for trout as the fish, but is not limited thereto.

According to the composition containing Blautia obeum, endoplasmic reticulum, lysate, culture medium or a mixture thereof according to one aspect as an active ingredient, the effect that can be usefully used for the prevention, improvement or treatment of degenerative brain diseases is there is.

1 is a graph showing Grip Strength when a strain according to one embodiment was administered to Parkinson's disease-induced mice.
Figure 2 is a graph showing the change in body weight when the strain according to one embodiment is administered to Parkinson's disease-induced mice.
Figure 3 is a graph showing changes in brain weight when the strain according to one embodiment is administered to Parkinson's disease-induced mice.

Hereinafter, the present invention will be described in more detail with examples, but this is only illustrative and is not intended to limit the scope of the present invention. It is obvious to those skilled in the art that the embodiments described below may be modified without departing from the essential gist of the invention.

Example 1. Preparation of strains

Blautia obeum deposited as KCTC14560BP was prepared as follows.

Specifically, in order to culture the strain, the strain was cultured in PYG broth (DSMZ 104) at 37° C. under anaerobic conditions for 2 days. Thereafter, the culture solution was centrifuged at 5000 x g for 20 minutes to remove bacterial debris. Thereafter, it was dispensed into clean PYG broth (DSMZ104) at 1x10^9 cfu/ml, stored in a cryogenic freezer, and used for experiments.

Example 2. Confirmation of the effect of preventing or improving degenerative brain diseases in animal models

In order to confirm the effect of preventing or improving degenerative brain disease of the strain according to one embodiment, an experiment was performed as follows.

Specifically, the mice were first subjected to a 7-day acclimatization period, and then orally administered the strain of Example 1 once a day for a total of 4 weeks around 2 weeks before and after Parkinson's disease was induced. After administration of the strain for 2 weeks, MPTP (1-Methyl-4-phenyl-1,2,3,6 tetrahydropyridine Hydrochloride) Parkinson's Disease inducer was orally administered to the strain on the 13th day at 20 mg/kg every 2 hours for a total of 4 It was administered intraperitoneally, and administered once subcutaneously on the 18th and 23rd days, respectively. Grip strength was evaluated on the 27th day of oral administration of the strain. Grip strength was measured by grabbing the mouse's tail and having all four limbs of the mouse hold the grid, and then pulling the mouse's tail with a constant force. A total of 3 times were performed and the average of the 3 times was obtained. Compared to the untreated group (WT) and the negative control group (MPTP) treated with only MPTP, the strain administration group ( B.obeum ) showed the effect.

As a result, as shown in Table 1 and FIG. 1, it was confirmed that the grip strength increased compared to the negative control group treated with only MPTP when the strain was administered according to one embodiment.

army WT MPTP B. obeum Average grip strength (N) 221.33 179.95 211.92

These results indicate that the strain according to one embodiment has an effect of improving symptoms according to degenerative brain disease.

Example 3. Confirmation of body weight and brain weight changes in animal models

When a Parkinson's disease-induced mouse and the strain according to one embodiment were administered thereto, changes in body weight and brain weight were confirmed as follows.

Specifically, the strain and MPTP according to one embodiment were administered in the same manner as in Example 2, and the body weight was measured a total of 5 times, once a week during the strain administration period, during group separation. Blood collection and necropsy were performed 24 hours after the last strain was administered. Blood was collected, separated from plasma, and frozen. After blood collection, individuals were perfused with 0.9% physiological saline to remove residual blood, and brains were removed and weighed.

As a result, as shown in Figures 2 and 3, it was confirmed that the body weight and brain weight did not change.

These results indicate that the strain according to one embodiment has an effect of improving degenerative brain disease without a change in body weight and brain weight.

Korea Center for Biological Resources (Overseas) KCTC14560BP 20210503

Claims (7)

Blautia obeum deposited under accession number KCTC 14560BP, endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or a pharmaceutical composition for the prevention or treatment of degenerative brain diseases containing a mixture thereof as an active ingredient As a composition, the degenerative brain disease is Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, ataxia, spinal cerebellar ataxia (Spinocerebellar Atrophy), Friedrich's Ataxia, A pharmaceutical composition selected from the group consisting of Machado-Joseph's disease, dystonia, and progressive supranuclear palsy. delete The pharmaceutical composition according to claim 1, wherein the Blautia obeum is live or dead bacteria. delete The method according to claim 1, wherein the composition is administered by an administration route selected from the group consisting of oral administration, intravenous administration, intraperitoneal administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration, and intrarectal administration. That will be, the pharmaceutical composition. Health function for preventing or improving degenerative brain diseases containing Blautia obeum deposited under accession number KCTC 14560BP, endoplasmic reticulum derived from the strain, lysate of the strain, culture medium, or a mixture thereof as an active ingredient food composition. Feed composition for preventing or improving degenerative brain disease comprising Blautia obeum deposited under accession number KCTC 14560BP, endoplasmic reticulum derived from the strain, lysate of the strain, culture medium, or a mixture thereof as an active ingredient .

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