KR20220038609A - Pharmaceutical Composition for Preventing or Treating Diabetes Comprising Heat Killed Akkermansia muciniphila - Google Patents

Abstract

The present invention provides a composition for preventing or treating diabetes containing a microorganism as an active ingredient. More specifically, the present invention provides a composition for preventing or treating diabetes in which the microorganism is Akkermansia muciniphila. The composition for preventing or treating diabetes of the present invention presents the concentration of Akkermansia muciniphila, which has the best anti-diabetic effect, thereby having excellent antidiabetic effect and being able to prevent unnecessary intake.

Description

A composition for preventing or treating diabetes comprising dead Akkermansia muciniphila {Pharmaceutical Composition for Preventing or Treating Diabetes Comprising Heat Killed Akkermansia muciniphila}

The present invention relates to a composition for preventing or treating diabetes comprising Akermansia muciniphila, and more particularly, in the composition for preventing or treating diabetes containing a microorganism as an active ingredient, wherein the microorganism is Akermansia mucini It relates to a composition for preventing or treating diabetes, which is a dead cell of Akkermansia muciniphila .

The number of diabetic patients in Korea increased from about 300,000 in the 1970s to about 4 million, or about 10% of the total population, in 2004. Worldwide, there were 110 million people in 1994, and it was 239 million in 2010, an increase of about 2.2 times, and it is one of the top 10 causes of death. Diabetes mellitus is caused by the generation of excessive free radicals that attack biological tissues and damage cells.

In particular, diabetic complications cause chronic metabolic diseases due to impaired metabolic control in the body due to increased peroxide production and decreased antioxidant enzyme activity. As a result, the epithelial cell membrane of capillary blood vessels thickens along with lipid metabolism abnormalities due to an increase in blood triglycerides, LDL-cholesterol and peroxide and a decrease in HDL-cholesterol, leading to cardiovascular disease, blindness due to retinal disease, stroke, myocardial infarction, and It is known as a disease characterized by high blood sugar, which causes many complications such as chronic renal failure. Because of these serious consequences, it cannot but be a national solution, such as a sharp increase in social security expenditures due to diabetes.

Therefore, prevention and delay of diabetes and its complications are the main goals of treatment. Although many studies have been conducted worldwide for the treatment and prevention of diabetes so far, the prevalence of diabetes is steadily increasing. In particular, type 2 diabetes, which accounts for more than 90% of diabetic patients, is known to be caused by a problem in insulin action or a defect in insulin secretion, and many oral hypoglycemic agents have been developed and used in clinical practice. Although it is possible to control blood sugar to some extent if oral hypoglycemic agents or insulin preparations are actively used, for successful treatment of type 2 diabetes, which has a more complex pathogenesis than type 1 diabetes, it is necessary to have two or three mechanisms instead of one. The development of pharmaceuticals is more demanded than ever.

However, oral hypoglycemic agents classified as sulfoneureas, biguanides and a-glucosidase inhibitors currently used for the treatment of diabetes cause hypoglycemia, abdominal pain, heart damage, and lacticacidosis in patients with impaired renal or cardiovascular function. It is accompanied by a number of side effects, such as In addition, the diabetes treatment market is showing a steady increase, but most of the raw materials or finished products are imported. It is judged that the development of functional foods for lowering blood sugar using food materials that have been found to be well-known can greatly contribute to the improvement of national health and quality of life as well as reduction of national medical expenses.

On the other hand, Akkermansia muciniphila has no history of being approved as a food or drug at home and abroad, and Professor Carney's research team at the University of Leuven in Belgium has a history of using live and dead Akermansia muciniphila in clinical trials on 54 patients. There is this.

According to the results of non-clinical and clinical tests, the level of inflammation in the body by inhibiting the passage of LPS (a major pro-inflammatory agent) through the intestinal lining by strengthening the gut barrier and improving metabolic disorders such as reducing body fat, improving lipids and increasing insulin sensitivity A number of studies have shown that it has a lowering effect. In addition, as a result of research conducted by Dr. Gitvogel of the Gustav Roussie Institute, it is confirmed that Akkermansia muciniphila activates T cells (T-Cell) in the intestinal mesentery to increase the reactivity of anticancer drugs (PD1/PDL1 inhibitors) (Bertrand Routy) et al., Gut microbiome influences efficacy of PD-1-based immunotherapy against epithelial tumors. Science, Vol. 359, Issue 6371, Jan 2018).

However, to date, the precise administration concentration of Acermansia muciniphila as a composition for preventing or treating diabetes has not been disclosed.

Numerous papers and patent documents are referenced throughout this specification and their citations are indicated. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to more clearly describe the level of the technical field to which the present invention pertains and the content of the present invention.

In existing studies on anti-diabetes, it is known that Acermansia muciniphila has anti-diabetic efficacy together with Bifidobacterium (e.g., Korean Patent Application Laid-Open No. 10-2017-0087540; in mucin-free medium A composition for the prevention, improvement or treatment of metabolic diseases containing the cultured Akermansia muciniphila strain or a culture medium thereof as an active ingredient), a domestic patent for a composition having a metabolic disorder treatment activity by Akkermansia muciniphila alone ( There is only published Patent 10-2015-7016539). Accordingly, the present inventors confirmed that there was an effect of inhibiting glucose absorption through a glucose tolerance test using dead Akermansia muciniphila. The present invention was completed by discovering that it was possible to more synergistically lower the blood sugar concentration.

Accordingly, an object of the present invention is to provide a composition for preventing or treating diabetes containing dead Akermansia muciniphila.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

The present invention provides a composition for preventing or treating diabetes.

In existing studies on anti-diabetes, it is known that Acermansia muciniphila has anti-diabetic efficacy together with Bifidobacterium (e.g., Korean Patent Application Laid-Open No. 10-2017-0087540; in mucin-free medium A composition for the prevention, improvement or treatment of metabolic diseases containing the cultured Akermansia muciniphila strain or a culture medium thereof as an active ingredient), a domestic patent for a composition having a metabolic disorder treatment activity by Akkermansia muciniphila alone ( There is only published Patent 10-2015-7016539). Accordingly, the present inventors confirmed that there was an effect of inhibiting glucose absorption through a glucose tolerance test using dead Akermansia muciniphila. It was confirmed that the blood sugar concentration could be lowered more synergistically.

According to one aspect of the present invention, in the composition for preventing or treating diabetes containing a microorganism as an active ingredient, the microorganism is Akkermansia muciniphila It provides a composition for preventing or treating diabetes .

As used herein, the term “diabetes” refers to a chronic disease characterized by a relative or absolute deficiency of insulin resulting in glucose-intolerance. The term diabetes includes all types of diabetes, including, for example, type 1 diabetes, type 2 diabetes and hereditary diabetes. Type 1 diabetes is insulin-dependent diabetes mellitus, mainly caused by destruction of β-cells. Type 2 diabetes is non-insulin-dependent diabetes mellitus, caused by insufficient insulin secretion after a meal or by insulin resistance.

The term 'health functional food' as used in the present invention refers to a food having bioregulatory functions such as disease prevention and treatment, body defense, immunity, recovery from illness, and aging suppression, and should be harmless to the human body when taken for a long time .

According to a preferred aspect of the present invention, the Akkermansia muciniphila of the present invention may preferably be a dead cell.

According to a preferred aspect of the present invention, the Akkermansia muciniphila of the present invention may be preferably freeze-dried (Freeze dried) or sterilized dried (Heat-killed) dead cells.

According to a preferred embodiment of the present invention, the mouse dose of the Acermansia muciniphila of the present invention may be preferably 2×10 ⁶ /200 ul/two to 2×10 ⁸ /200 ul/two, and most Preferably, it may be 2×10 ⁷ /200 ul/two.

According to a preferred aspect of the present invention, the human dose of the Akkermansia muciniphila of the present invention may be preferably 6.0-7.0×10 ⁴ /kg to 6.0-7.0×10 ⁶ /kg, most preferably may be 6.0-7.0×10 ⁵ /kg.

When converting the human/kg dose based on the medium concentration 2x10 ⁷ /25g (per subject) used in mice to 8x10 ⁸ /kg, 8x10 ⁸ /kg x 0.081 (body surface area of the mouse) = 6.48 x 10 ⁵ /human (See Anroop B. Nair et al., Journal of Basic and Clinical Pharmacy Vol. 7: 27-31, 2016).

When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. it's not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and is preferably applied by oral administration.

A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity of the patient. can be A preferred dosage of the pharmaceutical composition of the present invention is within the range of 0.001-100 mg/kg for adults.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by a person of ordinary skill in the art to which the present invention pertains. or it may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or an aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.

When the composition of the present invention is prepared as a food composition, it includes not only juniferanol as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. includes the Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared as a drink, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, licorice root extract, jujube extract, licorice extract, etc. may be additionally included in addition to the juniferanol of the present invention. can

According to another aspect of the present invention, the present invention provides a health functional food for preventing or treating diabetes containing a microorganism as an active ingredient, wherein the microorganism is Akkermansia muciniphila for preventing or treating diabetes mellitus Provides functional foods.

In the present invention, matters related to health functional food for preventing or treating diabetes will be omitted in detail in order to prevent the present specification from becoming excessively complicated in relation to the aforementioned composition for preventing or treating diabetes.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a composition for preventing or treating diabetes containing a microorganism as an active ingredient, wherein the microorganism is Akkermansia muciniphila .

(b) The composition for preventing or treating diabetes according to the present invention provides the highest concentration of Akkermansia muciniphila, which has the highest antidiabetic effect, and thus has excellent antidiabetic efficacy as well as the advantage of preventing unnecessary intake.

1 shows the results of analysis of the oral glucose tolerance of Akkermansia muciniphila (Control: DW, Media: Media 200 ul/doo, AML: Akkermansia muciniphila 2×10 ⁶ /200 ul/head, AMM: Akkermansia muciniphila 2×10 ⁷ /200 ul/head, AMH: Akkermansia muciniphila 2×10 ⁸ /200 ul/head, the statistical significance level of the mean value for each experimental group is indicated by each subgroup for p<0.05).
Figure 2 shows the AUC (area under the curve) analysis results of Akkermansia muciniphila (Control: DW, Media: Media 200 ul / two, AML: Akkermansia muciniphila 2 × 10 ⁶ /200 ul / two, AMM: Akkermansia muciniphila 2×10 ⁷ /200 ul/head, AMH: Akkermansia muciniphila 2×10 ⁸ /200 ul/head, the statistical significance level of the mean value for each experimental group is indicated by each subgroup for p<0.05)

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it is for those of ordinary skill in the art to which the present invention pertains that the scope of the present invention is not limited by these examples according to the gist of the present invention. it will be self-evident

Example

Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight) % for solid/solid, (weight/volume) % for solid/liquid, and Liquid/liquid is (volume/volume) %.

test material

Akermansia muciniphila is a strain that corresponds to Level 1 (BL1) in the Biosafety Level (BL) standards established by the Centers for Disease Control (CDC) and National Institutes of Health (NIH), which have sufficient characteristics. It is known that it is not harmful to the environment or experimenter, is not pathogenic, and does not cause disease in healthy people (American Type Culture Collection (ATCC) certification: BAA-835TM (Akkermansia muciniphila)).

The blank samples were supplied by Wooridoo Co., Ltd., which will sign a production agreement with Hanteo Co., Ltd., and a blood glucose improvement screening evaluation was conducted using a total of two blank samples. For animal testing, a bacterial culture medium was used for dilution, and it was used under low-temperature storage.

test code test substance import quantity appearance AM Acamancia muciniphila
( Akkermansia muciniphila ) dead cells E-tube
24 ea solid/powder Media culture medium 4 bottles Liquid

Test Methods

Test animals and breeding environment

ICR mice (6 weeks old, male), an experimental animal, were purchased from Orient Bio (Gyeonggi, Korea) and used in the experiment after acclimatization for 7 days. Each experimental group was divided into a control group and each test group (Ackermansia muciniphila concentration-specific test group or medium test group), and the experiment was conducted by randomly placing 10 heads each. For the experimental diet, general solid feed (Samtako, Gyunggi, Korea) was fed, and during the acclimatization period, the filtered drinking water was changed daily to allow free intake. During breeding period, the temperature is 23±1℃, humidity 50±5%, noise 60 phone or less, lighting time 08:00~20:00 (12 hours a day), illumination 150~300 Lux, ventilation 10~12 times per hour The meeting environment was maintained.

Oral glucose tolerance analysis test

Experimental animals that have been fasted for more than 8 hours have fasting blood glucose measured in the caudal vein using a blood glucose meter (AutoCheck, Diatech Korea), then randomly placed into groups using the egg mass method, and each sample is orally administered according to the dose. did After 30 minutes, blood glucose was measured again, and according to the experimental method, 2 g/kg of a blood pressure increasing factor (glucose) was administered to all groups. Thereafter, the area of the blood sugar change curve was obtained and the blood sugar change for each experimental group was analyzed, and the results are shown in FIG. 1 .

statistical analysis

All experimental results were calculated as mean ± standard error (Mean ± S.E.) using a statistical program (SPSS ver.12.0, SPSS Inc., Chicago, IL, USA). Statistical analysis according to the statistical significance test between each experimental group was carried out after ANOVA (one-way analysis of variance test).

Test result

Test Result 1: Analysis of Oral Glucose Tolerance of Acermansia muciniphila

In a normal state, blood sugar rises temporarily after a meal, but falls back to a normal level by insulin. However, if the pancreas does not secrete insulin well, or if the pancreas does not function properly, the blood sugar does not go down to a normal level after a meal. In other words, the body cannot use glucose as an energy source, but excretes it. If the blood sugar level is higher than normal, it may interfere with many regulators transported through the blood, decrease the functions of red blood cells (oxygen transport) and white blood cells (clean blood vessels), or put a strain on the kidneys, which has adverse effects on our body. will give Accordingly, in the present invention, the effect of the culture medium and dead cells of Acermansia muciniphila on the blood glucose change due to glucose administration was investigated. As a result of this experiment, the blood glucose level after glucose administration (30 min) was 334.86±19.96 mg/dL in the control group, 309.29±28.81 mg/dL in the Media group, and 277.71±8.95 mg/dL in the low concentration group (AML). , the medium concentration group (AMM) was 252.29±12.69 mg/dL, and the high concentration group (AMH) 314.43±22.19 mg/dL. In particular, it was observed that the blood glucose level of the medium concentration administration group was significantly lower than that of the control group (see FIG. 1 ).

Test Result 2: AUC (area under the curve) analysis of Ackermansia muciniphila

Among the methods of measuring blood glucose, glucose tolerance measures the blood glucose concentration for each hour as a line on the graph and compares the area (AUC, incremental area under the curve) of the graph that appears between 0-180 minutes. AUC (Area under the curve) analysis was also similar to the previous results, and the change in blood glucose in the test group for glucose was 27778.00±1438.01 mg/dL in the control group, 24765.49±1654.82 mg/dL in the Media group, low concentration. It was found to be 24800.23±990.04 mg/dL in the administration group (AML), 23210.71±1244.54 mg/dL in the medium dose group (AMM), and 25013.41±1104.03 mg/dL in the high dose group (AMH). The blood glucose change of the control group was the highest, whereas all of the test groups were observed to be significantly lower. In particular, the medium concentration administration group was observed to be the most significantly lower.

conclusion

Glucose absorbed into the body is burned in peripheral tissues, or blood sugar can be controlled by stimulating beta cells to secrete insulin. As such, the ability of the body to metabolize glucose normally is called glucose tolerance.

Even though they do not fall under the category of diabetes, people with high blood sugar often present with impaired glucose tolerance (IGT). Conventionally, diabetes has been referred to as a case where the concentration of glucose in venous blood is 15 mmol/l (270 mg/dl) or more, but the WHO has more standardized guidelines ① The fasting blood glucose concentration is 7.8 mmol/l (140 mg/dl). Diabetes is defined as diabetes when the blood glucose concentration is 11.1 mmol/l (200 mg/dl) or more after 2 hours of oral glucose tolerance (OGT) of 75 glucose. In contrast, IGT refers to when the blood glucose concentration is between 7.8-11.1 mmol (140-200mg/dl) after 2 hours in the 75g-OGT test.

Hyperglycemia may be due to this decrease in glucose tolerance. Therefore, in order to verify the glycemic control effect of functional foods, glucose tolerance tests on individuals are essential.

In the screening evaluation, the test group administered with Akkermansia muciniphila was found to be effective in suppressing glucose-induced rise in blood sugar compared to the control group, of which 2×10 ⁷ /200 ul/two It was investigated that the blood sugar control effect was the highest in the medium concentration test group administered with .

Accordingly, Akkermansia muciniphila of this material is thought to have an effect on blood sugar reduction, and it is thought that functional raw material registration is possible through additional research (test before human application, test for human application).

Claims (10)

In the composition for preventing or treating diabetes containing a microorganism as an active ingredient,
The microorganism is Akkermansia muciniphila ( Akkermansia muciniphila )
A composition for preventing or treating diabetes. The method of claim 1,
The Akkermansia muciniphila ( Akkermansia muciniphila ) is a composition for preventing or treating diabetes, characterized in that it is a dead cell. 3. The method of claim 2,
The Akkermansia muciniphila (Akkermansia muciniphila) is a composition for preventing or treating diabetes, characterized in that the freeze-dried (Freeze dried) or sterilized dried (Heat-killed) dead cells. 4. The method of claim 3,
The human dose of Akkermansia muciniphila is 6.0-7.0×10 ⁴ /kg to 6.0-7.0×10 ⁶ /kg of diabetes preventing or treating composition. 5. The method of claim 4,
The human dose of Akkermansia muciniphila is 6.0-7.0×10 ⁵ /kg of diabetes preventing or treating composition. In the health functional food for preventing or treating diabetes containing microorganisms as an active ingredient,
The microorganism is Akkermansia muciniphila ( Akkermansia muciniphila )
Health functional food for preventing or treating diabetes. 7. The method of claim 6,
The Akkermansia muciniphila ( Akkermansia muciniphila ) is a diabetes preventive or health functional food, characterized in that the dead cells. 8. The method of claim 7,
The Akkermansia muciniphila (Akkermansia muciniphila) is a health functional food for preventing or treating diabetes, characterized in that the freeze-dried (Freeze dried) or sterilized dried (Heat-killed) dead cells. 9. The method of claim 8,
The human dose of Akkermansia muciniphila is 6.0-7.0×10 ⁴ /kg to 6.0-7.0×10 ⁶ /kg of diabetes prevention or treatment health functional food. 10. The method of claim 9,
The human dose of Akkermansia muciniphila is a health functional food for preventing or treating diabetes, characterized in that 6.0-7.0×10 ⁵ /kg.

Images