WO2023219460A1 - Composition comprising lactobacillus fermentum strain and metabolism modulator for preventing or treating metabolic diseases by using combination therapy - Google Patents

Abstract

The present invention relates to a metabolic disease treatment agent in which a Lactobacillus fermentum strain and a metabolism modulator are co-administered. The Lactobacillus fermentum strain according to one embodiment exhibits anti-obesity activity and the activity of reducing side effect symptoms of the metabolic modulator, and, particularly, a composition or a health functional food for treating metabolic diseases, in which the Lactobacillus fermentum strain and the metabolic modulator are co-administered, can be effectively used for preventing or treating metabolic diseases.

Description

Composition for preventing or treating metabolic disease using combination therapy containing Lactobacillus fermentum strain and metabolic regulator

This relates to a treatment for metabolic diseases administered in combination with a Lactobacillus fermentum strain and a metabolic regulator.

Obesity refers to a state in which excessive fat tissue is accumulated in the body due to an increase in the size or number of fat cells, that is, a state in which there is more fat than the body needs. Obesity is considered the first correctable cause of death among modern people, and is a chronic disease that must be managed to enjoy a healthy and vibrant life. It is known that obesity is associated with various diseases such as high blood pressure, diabetes, hyperlipidemia, angina, fatty liver, cancer, and degenerative arthritis. As metabolic diseases caused by obesity become more diverse and their severity worsens, prevention and treatment are urgently needed to prevent them.

As the number of obese patients increases significantly around the world, global companies researching obesity treatments are accelerating their development. In particular, as the novel coronavirus infection (Corona 19) situation continued for more than two years, it was found that the number of obese patients increased rapidly during this period due to a decrease in activity.

While health authorities in each country are also concerned about the growing social and economic losses due to the increase in the obesity population, the obesity treatment market is expected to grow rapidly as demand increases.

Currently, many obesity treatments are being developed and commercialized as products. However, the appetite suppressant class of obesity treatments has the advantage of excellent weight loss efficacy, but these appetite suppressant class obesity treatments have a mechanism of action that reduces food intake by acting on the central nervous system to reduce appetite, and thus have a long-term effect. There is a critical problem that the effect does not last long or causes serious side effects during long-term use. As treatments based on GLP-1 analogues have been developed and commercialized to effectively treat obesity without side effects, they are receiving a lot of attention in the obesity market.

GLP-1 is a representative gastrointestinal hormone and neurohormone that is involved in regulating blood sugar levels following food intake. GLP-1 is a hormone secreted from the small intestine stimulated by food intake. It promotes insulin secretion from the pancreas in a blood sugar concentration-dependent manner and helps lower blood sugar concentration by suppressing the secretion of glucagon. Additionally, it acts as a satiety factor and reduces food intake. Accordingly, it has been reported that it has the effect of suppressing food intake and reducing weight in both normal and obese states, showing its potential as a treatment for obesity.

Research is actively underway to develop a treatment for diabetes and obesity using the blood sugar control and weight loss effects of GLP-1, and a representative example is Novo Nordisk's Liraglutide. Liraglutide is a long-acting GLP-1 receptor agonist that binds to the same receptor as endogenous GLP-1 and promotes insulin secretion, thereby regulating blood sugar and reducing appetite, suppressing weight gain and lowering triglycerides (Saxenda). It was released in the United States and Europe under the name (Expert Rev Cardiovasc Ther. 2015;13(7):753-767). However, side effects of these GLP-1 receptor agonists have also been reported, including nausea, vomiting, decreased appetite, headache, constipation, and abdominal bloating (Korean J Med. 2014;87(1):9-13).

Accordingly, in order to increase the treatment effect and satisfaction of patients, there is a need for research to maintain or improve the effects of GLP-1 administration, such as blood sugar control and weight loss, and to alleviate side effects caused by GLP-1 administration.

One aspect includes as an active ingredient a first active substance comprising a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator. do or; Alternatively, it provides a pharmaceutical composition for preventing or treating metabolic diseases, which includes the first active substance as an active ingredient and wherein the second active substance is administered in combination.

Another aspect includes the first active agent and the second active agent as active ingredients; Alternatively, a health functional food for preventing or improving metabolic disease is provided, which includes the first active substance as an active ingredient and the second active substance is administered in combination.

Another aspect includes the first active material and the second active material as active ingredients; Alternatively, a kit for preventing or treating metabolic diseases is provided, which includes the first active substance as an active ingredient and the second active substance is administered in combination.

Another aspect includes the first active material and the second active material as active ingredients; Alternatively, it provides a method of delivering a drug into a subject comprising administering to an subject in need a composition containing the first active substance as an active ingredient and administering the second active substance in combination.

Another aspect includes the first active material and the second active material as active ingredients; Alternatively, a method for preventing or treating metabolic diseases is provided, comprising administering to an individual in need a composition containing the first active substance as an active ingredient and administering the second active substance in combination.

Another aspect includes the first active material and the second active material as active ingredients; or a composition for preventing or treating metabolic diseases comprising the first active substance as an active ingredient and in which the second active substance is administered in combination; Or it provides use in the manufacture of a health functional food composition for preventing or improving metabolic diseases.

Another aspect provides a composition for preventing or treating symptoms caused by side effects of a metabolic regulator containing Lactobacillus fermentum strain as an active ingredient.

Another aspect provides a health functional food for preventing or improving symptoms caused by side effects of a metabolic regulator containing Lactobacillus fermentum strain as an active ingredient.

Another aspect is a method for preventing or treating symptoms caused by side effects of a metabolic regulator by administering a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof to an individual in need thereof. provides.

Another aspect is a composition for preventing or treating symptoms caused by side effects of a metabolic regulator using a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof; Alternatively, it is provided for use in the manufacture of a health functional food composition for preventing or improving symptoms caused by side effects of metabolic regulators.

One aspect provides a Lactobacillus genus strain, specifically a Lactobacillus fermentum strain, which has anti-obesity activity or activity to alleviate the side effects of metabolic regulators.

Lactobacillus is a genus of aerobic or facultative anaerobic Gram-positive bacilli widely distributed in nature. Microorganisms belonging to the Lactobacillus genus include Lactobacillus fermentum, Plantarum, and Sakei. As a result of research to develop a new strain, the present inventors selected Lactobacillus fermentum GB102 as a candidate strain with anti-obesity activity. The strain was deposited at the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center under deposit number SD1335 on September 6, 2019. The same strain described above was deposited at the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center under the deposit number KCTC14105BP on January 14, 2020. The strain corresponds to a probiotic strain, is harmless to the human body, and can be used without side effects.

The Lactobacillus ( Lactobacillus ) has been renamed to Limosilactobacillus or Lactiplantibacillus , and the changed strain names in this specification can be used interchangeably. For example, Lactobacillus fermentum was changed to Limosilactobacillus fermentum .

As used herein, the term “ Lactobacillus fermentum GB102” may be interchangeably described with L. fermentum GB102 strain, Lactobacillus fermentum GB102 strain, or GB102.

In one embodiment, the strain may be a strain deposited under accession number KCTC14105BP.

In one embodiment, the strain may be a strain having 16S rRNA comprising the nucleotide sequence of SEQ ID NO: 1 or a 16s rRNA comprising a nucleotide sequence having at least 98.5% nucleotide sequence identity. Specifically, it has at least 93%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% homology with the nucleotide sequence consisting of SEQ ID NO: 1 of the present specification.

In one embodiment, the strain may be a strain containing a 16S rRNA gene consisting of the nucleotide sequence of SEQ ID NO: 1.

In one embodiment, the strain may be a strain that is a mutation of a naturally occurring strain.

In one embodiment, the strain may be live cells, dead cells, or a cytoplasmic fraction obtained by disrupting the strain, and preferably may be live cells.

In one embodiment, the strain may have anti-obesity activity. Specifically, the anti-obesity activity inhibits weight gain, improves glucose or insulin resistance, increases energy metabolism or basal metabolic rate, accumulates brown fat, reduces body fat mass compared to lean body mass, suppresses liver steatosis, and increases blood AST (alanine). aminotransferase), ALT (alanine transaminase, GPT), reducing triglycerides, low-density cholesterol, high-density cholesterol or total cholesterol, and increasing the expression of UCP-1 (Uncoupling protein 1). You can. Therefore, the strain may inhibit weight gain or improve blood sugar control or glucose tolerance.

In one embodiment, the strain may have the activity of alleviating the side effects of metabolism regulators. Specifically, the metabolic regulator, for example, GLP-1 (Glucagon-like peptide-1) receptor agonist, when administered, improves body weight, blood pressure, and glycated hemoglobin levels in obese patients, improves insulin sensitivity, or improves glycated hemoglobin levels in obese patients. Blood sugar control is improved, but when administration is stopped, body weight, blood pressure, glycated hemoglobin, and blood sugar control return to the values prior to administration of the metabolic regulator or worsen further side effects. The strain may alleviate or improve the side effects of the metabolism regulator, specifically, increase in body weight, increase in blood pressure, worsening of blood sugar control ability, and increase in glycated hemoglobin after discontinuation of administration of the metabolism regulator. Therefore, the strain is not directly involved in the level or expression of metabolic regulators, specifically GLP-1, but improves the tolerance of metabolic regulators, specifically GLP-1, to reduce the symptoms of side effects caused by taking GLP-1. This may be to improve the efficacy of GLP-1 by alleviating or increasing the sensitivity of GLP-1.

As used herein, the term “culture” may be used interchangeably with “culture supernatant,” “culture supernatant,” “conditioned culture,” or “conditioned medium,” and can be used interchangeably with Lactobacillus strains to grow and survive in vitro. It may refer to the entire medium containing the strain, its metabolites, extra nutrients, etc. obtained by culturing the strain in a medium capable of supplying nutrients for a certain period of time. The culture refers to a product obtained by culturing a probiotic strain in a known medium, and the product may or may not include the strain itself. The medium may be selected from known liquid media or solid media, for example, MRS liquid medium, GAM liquid medium, MRS agar medium, GAM agar medium, and BL agar medium, but is not limited thereto.

As used herein, the term “lysate” may be used interchangeably with “lysate”, meaning a solution or suspension in an aqueous medium of cells of a microorganism, such as broken Lactobacillus fermentum. Cell lysates include, for example, macromolecules such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc. and/or micromolecules such as amino acids, sugars, fatty acids, etc., or fractions thereof. The lysate also contains cell debris, which may be smooth or granular in structure.

The culture medium may include the culture medium itself, its concentrate, or freeze-dried product obtained by cultivating the strain, or the culture supernatant obtained by removing the strain from the culture medium, its concentrate, or freeze-dried product.

The culture medium may be obtained by culturing Lactobacillus fermentum in an appropriate medium (e.g., MRS plate medium) at any temperature above 10°C or below 40°C for a certain period of time, for example, 4 to 50 hours. The culture medium and culture conditions for cultivating the Lactobacillus fermentum can be appropriately selected or modified by those skilled in the art.

Another aspect includes a first active substance comprising a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator as the active ingredient. do or; Alternatively, a composition is provided that includes the first active substance as an active ingredient and the second active substance is administered in combination.

In one embodiment, the first active substance may include succinic acid.

The succinic acid may be represented by the following formula (1):

[Formula 1]

The succinic acid is administered to a subject and induces the accumulation of brown fat in the subject's fat cells. In mammals, there are two types of adipose tissue: white adipose tissue and brown adipose tissue (Gesta S et al., Cell 2007;131:242-256). Additionally, beige adipose tissue is known to contribute to the browning of white fat. The white adipose tissue is used by the individual for energy storage and therefore its excess is associated with obesity. In contrast, brown adipose tissue has a high concentration of mitochondria and can uniquely express uncoupling protein 1 (UCP1). Therefore, brown adipose tissue can play a role in improving obesity by increasing energy consumption and thermogenesis. Therefore, the first active substance containing succinic acid has anti-obesity activity by increasing energy metabolism or basal metabolic rate through accumulation of brown fat when administered to a subject.

In one embodiment, the anti-obesity activity includes inhibiting body weight gain, improving glucose or insulin resistance, increasing basal metabolic rate, accumulating brown fat, reducing body fat mass compared to lean body mass, suppressing liver steatosis, and increasing blood AST (alanine). aminotransferase), ALT (alanine transaminase, GPT), reducing triglycerides, low-density cholesterol, high-density cholesterol or total cholesterol, and increasing the expression of UCP-1 (Uncoupling protein 1). You can.

In one embodiment, the first active substance may include Lactobacillus fermentum strain alone as an active ingredient, or may include one or more pharmaceutically acceptable carriers, excipients, or diluents.

The metabolic regulator may include an incretin agent.

The metabolic regulator includes GLP-1 (Glucagon-like peptide-1) receptor, GLP-1 receptor agonist and analogues thereof, GIP (Glucose-dependent insuliontropic polypeptide) receptor, GIP receptor agonist and analogues thereof, glucagon agonist and analogues thereof, GLP -1/GIP dual receptor agonist, GLP-1/glucagon dual receptor agonist, and GLP-1/GIP/glucagon triple receptor agonist, DPP-4 inhibitor (Dipeptidyl peptidase-4 inhibitor) and its analogs, SGLT-2 inhibitor (Sodium - It may be one or more of Glucose Cotransporter 2 inhibitor) and its analogues, Oxyntomodulin agonist and its analogues, and Fibroblast Growth Factor-21 (FGF21) and its analogues. In one embodiment, the metabolism regulator may be a GLP-1 receptor agonist or an analog thereof.

Specifically, GLP-1 receptor agonists and analogs thereof include Exenatide, Lixisenatide, Liraglutide, Albiglutide, Taspoglutide, and Sema. It includes Semaglutide, Dulaglutide, and Orforglipron (LY3502970), but the type of GLP-1 receptor agonist of the present invention is not limited thereto and is a general purpose agent with the same or similar effect. Any agent that can be used can be used. In one embodiment, the metabolism regulator may be dulaglutide.

Specifically, GLP-1/GIP dual receptor agonists include Tirzepatide, NN9709, MAR709, RG7697, AMG 133, VK2735, and SAR-438335, but the types of GLP-1/GIP dual receptor agonists of the present invention is not limited to this, and general-purpose agents with the same or similar effects can be used.

Specifically, GLP-1/glucagon dual receptor agonists include Cotadutide, Pemvidutide, ALT 801, SP 1373, VPD-107, BI456906, SAR425889, HM12525, DA-1726, and ZP2929. , the type of GLP-1/glucagon dual receptor agonist of the present invention is not limited to this, and general-purpose agonists having the same or similar effects can be used.

Specifically, GLP-1/GIP/glucagon triple receptor agonists include Efinopegdutide, efocipegtrutide, Retatrutide, HM 15211, LY3437943, and SAR441255; The type of GLP-1/GIP/glucagon triple receptor agonist of the present invention is not limited to this, and general-purpose agonists having the same or similar effects can be used.

Specifically, the GLP-1/FGF21 dual receptor agonist includes, but is not limited to, YH25724, and a general agonist having the same or similar effect may be used.

Additionally, DPP-4 inhibitors and their analogs include Linagliptin, Sitagliptin, Vildagliptin, Saxagliptin, denagliptin, and Alogliptin. ), Carmegliptin, Melogliptin, Gosogliptin, Teneligliptin and Dutogliptin, SGLT-2 inhibitors and their analogs are Including, but not limited to, Dapagliflozin, Canagliflozin, ertugliflozin, Remogliflozin, and Sergliflozin. That is not the case.

In one embodiment, the second active substance may contain a metabolic regulator or a metabolic peptide as an active ingredient alone, or may be provided as a pharmaceutical composition including one or more pharmaceutically acceptable carriers, excipients, or diluents. .

Another aspect is to provide Lactobacillus fermentum strains, cultures of the strains, lysates of the strains, or mixtures thereof for the purpose of improving, preventing, or treating symptoms caused by side effects of metabolic regulators.

Another aspect is that a first active substance containing a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator are used as active ingredients. Contains; Alternatively, the composition includes the first active substance as an active ingredient, and the second active substance is administered in combination to provide a use for disease improvement, prevention or treatment.

As used herein, the term “treat” may mean curing metabolic diseases, etc. in a shorter time compared to natural healing. The treatment may include improvement and/or alleviation of metabolic disease. Additionally, the treatment may mean healing and/or recovery of symptoms caused by metabolic disease.

As used herein, the term “prevention” refers to partially or completely delaying or preventing the onset or recurrence of a disease, disorder, or its secondary symptoms, preventing the acquisition or re-acquisition of a disease or disorder, or preventing the development or recurrence of a disease or disorder. It refers to a method of reducing the risk of acquisition. For example, the above prevention refers to all actions that suppress or delay the occurrence of metabolic disease by administering the composition according to the present invention.

As used herein, the term "included as an active ingredient" means that a Lactobacillus fermentum strain, an endoplasmic reticulum derived from the strain, a lysate of the strain, a culture medium, or an extract of the culture medium, or a metabolic regulator is added, and drug delivery. This means that it is formulated into various forms by adding various ingredients as sub-ingredients for stabilization, etc.

The use of the composition is to address the side effects of metabolic regulators, specifically, when administered with a GLP-1 (Glucagon-like peptide-1) receptor agonist, body weight, blood pressure, and glycated hemoglobin levels are improved in obese patients, or blood sugar control is improved in diabetic patients. However, when administration is discontinued, body weight, blood pressure, glycated hemoglobin, and blood sugar control ability may return to the values prior to administration of the metabolic regulator, or may include improvement in symptoms through prevention, improvement, or treatment of worsening symptoms.

Uses of the composition may include improving the symptoms of metabolic disease, specifically preventing, improving, or treating obesity, diabetes, or insulin resistance.

As used herein, the term “obesity” refers to a state of excessive body fat. Clinically, obesity may be a case where the body mass index (BMI) is 25 in Korea and 30 or more according to the World Health Organization (WHO). In general, obesity means that your body weight is higher than normal, but even if you do not weigh a lot, you may be obese if your body fat ratio is high among the body components. The obesity can occur in both adults and children. The obesity is not only caused by weight gain, but also by overeating, overdrinking and bulimia, high blood pressure, diabetes, increased plasma insulin concentration, insulin resistance, hyperlipidemia, metabolic syndrome, insulin resistance syndrome, obesity-related gastroesophageal reflux, arteriosclerosis, hypercholesterolemia, and uric acid. It can cause obesity-related diseases such as hyperemia, cardiac hypertrophy and left ventricular hypertrophy, lipodystrophy, nonalcoholic steatohepatitis, cardiovascular disease, or polycystic ovary syndrome. Therefore, the composition can be used for preventing or treating not only obesity but also obesity-related diseases. Additionally, the composition can be used in individuals who have a desire to lose weight even if they are not obese.

The obesity may be due to various causes. For example, the causes may be a high-fat diet, reduced amount of exercise, genetics, psychological factors, endocrine system abnormalities, metabolic abnormalities, and social and environmental factors. In particular, the obesity may be induced by a high-fat diet.

In this specification, the term “metabolic disease” is a general term for various disorders that occur due to abnormalities in the metabolic process in the body, and is also called metabolic disorder. It is generally caused by imbalances in carbohydrates, lipids, proteins, vitamins, electrolytes, and moisture. Specifically, the metabolic diseases include, but are not limited to, metabolic syndrome, type 1 diabetes, type 2 diabetes, hypertension, hyperlipidemia, obesity, fatty liver, insulin resistance, coronary artery sclerosis, and arteriosclerosis.

As used herein, the term “insulin resistance” refers to the inability of insulin to lower blood sugar levels and the inability of cells to effectively burn glucose. When insulin resistance is high, the body produces too much insulin, which can cause high blood pressure or dyslipidemia, as well as heart disease and diabetes. In addition, insulin is regulated by insulin receptor, insulin-like growth factor receptor and/or insulin receptor substrate protein in cells and tissues.

According to one embodiment of the present invention, the metabolic disease is a group consisting of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, impaired fasting blood sugar, dyslipidemia, lipid metabolism disorder, obesity, fatty liver, insulin resistance syndrome, and glucose tolerance syndrome. It may be selected from .

In one embodiment, the anti-obesity activity of the first active substance or the activity of alleviating the side effects of metabolism regulators may be more effective when administered in combination with the second active substance.

As used herein, the term “combination therapy” or “combined treatment” or “in combination” refers to any form of simultaneous or parallel treatment using at least two separate therapeutic agents. The components of the combination therapy may be administered simultaneously, sequentially, in reverse order, or in any order. The components may be administered in any suitable manner, in different doses or at different frequencies of administration or via different routes.

As used herein, the term "administration" means introducing a predetermined substance into an individual by an appropriate method, and "individual" refers to all rats, mice, livestock, etc., including humans, that may have metabolic diseases or side effects of metabolic regulators. It means living things. As a specific example, it may be a mammal, including humans.

As used herein, the term “administered simultaneously” is not particularly limited and means that the components of the combination therapy are administered substantially simultaneously, for example as a mixture or in an immediately following sequence.

As used herein, the term “sequentially administered” is not particularly limited and means that the components of the combination therapy are not administered simultaneously, but are administered one by one or in clusters with a specific time interval between administrations. The time interval may be the same or different between the respective administrations of the components of the combination therapy and may be selected, for example, in the range of 2 minutes to 96 hours, 1 day to 7 days or 1 week, 2 weeks or 3 weeks. . Generally, the time interval between administrations can range from minutes to hours, for example from 2 minutes to 72 hours, 30 minutes to 24 hours, or 1 to 12 hours. Additional examples include time intervals ranging from 24 to 96 hours, 12 to 36 hours, 8 to 24 hours, and 6 to 12 hours.

The combined administration may mean administering the Lactobacillus fermentum strain and the metabolic regulator simultaneously, sequentially or individually, and in any order. Specifically, the combined administration may involve simultaneously administering a first active substance containing a Lactobacillus fermentum strain and a second active substance containing a metabolism regulator, or administering a metabolism regulator after administering the Lactobacillus fermentum strain. there is. The combination therapy according to the present invention is an efficacy that can be obtained by administering one or the rest of the components of the combination therapy at a conventional dose, for example, the efficacy measured through the degree of response, response rate, time to disease progression, or survival time. More therapeutically superior can be defined as being able to provide synergistic effects. For example, if the therapeutic efficacy is superior to the efficacy obtained by using each of the above alone, the efficacy of the combination treatment is synergistic. In particular, without harming one or more of the degree of response, rate of response, time to disease progression, and survival data, and in particular without harming the duration of response, than would occur when each component is used at its usual dose. , a synergistic effect is considered to exist if the typical dose of Lactobacillus fermentum strain and metabolic regulator can be reduced while problematic side effects are reduced and/or reduced.

In one embodiment, the composition may be a pharmaceutical composition.

In the pharmaceutical composition, the “strain”, “first active substance”, “metabolism regulator”, “second active substance”, “co-administration”, etc. may be within the above-mentioned range.

As used herein, the term “pharmaceutically acceptable” means that the composition exhibits non-toxic properties to cells or humans exposed to the composition.

Types of pharmaceutically active ingredients that can deliver the active ingredient into an individual include contrast agents (dyes), hormones, antihormones, vitamins, calcium agents, mineral agents, saccharides, organic acid agents, protein amino acid agents, detoxifiers, enzyme agents, Metabolic agents, diabetes mellitus, tissue rejuvenation agents, chlorophyll agents, dye agents, radiopharmaceuticals, tissue cell diagnostic agents, tissue cell therapeutic agents, antibiotic agents, antiviral agents, combination antibiotic agents, chemotherapy agents, vaccines, toxins, toxoids , antitoxins, leptospira serum, blood products, biological agents, analgesics, immunogenic molecules, antihistamines, allergy medications, non-specific immunogenic agents, anesthetics, stimulants, psychotropic agents, small molecule compounds, nucleic acids, aptamers, antisense nucleic acids, oligonucleotides. , peptides, siRNA, and micro RNA.

The composition may additionally include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, or a combination thereof. The carrier may be an excipient, disintegrant, binder, lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium monohydrogen phosphate anhydride, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

When the pharmaceutical composition is formulated, it may be prepared using commonly used diluents or excipients such as lubricants, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, fillers, extenders, binders, wetting agents, disintegrants, and surfactants. You can. Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc., and these solid preparations may contain at least one excipient, such as starch, calcium carbonate, or sucrose. ) or it can be prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used, and when manufacturing in the form of eye drops, known diluents or excipients can be used. there is.

The pharmaceutical composition contains carbohydrates such as glucose, sucrose or dextran, antioxidants such as Ascorbic acid or Glutathione, chelating agents, and small molecules to increase stability or absorption. Proteins or other stabilizers can be used as agents.

In one embodiment, the pharmaceutical composition may be formulated as an oral or parenteral dosage form. Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrup, or combinations thereof. For parenteral administration, you can choose the injection method by external injection under the skin, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

In one embodiment, the route of administration of the pharmaceutical composition is, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral. , topical, sublingual, or rectal. Specifically, the first active substance containing the Lactobacillus fermentum strain and the second active substance containing a metabolism regulator are administered orally in the form of granules, powders, solutions, tablets, capsules, dry syrup, or a combination thereof, or for external use on the skin or It can be administered parenterally by intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

In one embodiment, it includes a first oral formulation containing the first active material, and a second oral formulation containing the second active material, and the first and second oral formulations may be administered orally. In one embodiment, the first and second active substances can be administered orally in a single oral preparation.

The pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type and severity of the patient's disease, the activity of the drug, and the drug's effect. It can be determined based on factors including sensitivity, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

The composition according to one embodiment may include 0.001% by weight to 80% by weight of Lactobacillus fermentum strain based on the total weight of the composition. Additionally, the administered dose of the Lactobacillus fermentum strain may be 0.01 mg to 10,000 mg, 0.1 mg to 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg. The strain is included in the composition in a therapeutically effective amount or nutritionally effective concentration, for example, the strain is 10 ³ to 10 ¹⁶ CFU/g, 10 ³ to 10 ¹⁵ CFU/g, 10 ³ to 10 ¹⁴ CFU/g. g, 10 ³ to 10 ¹³ CFU/g, 10 ³ to 10 ¹² CFU/g, 10 ⁴ to 10 ¹⁶ CFU/g, 10 ⁴ to 10 ¹⁵ CFU/g, 10 ⁴ to 10 ¹⁴ CFU/g, 10 ⁴ to 10 ¹³ CFU/g, 10 ⁴ to 10 ¹² CFU/g, 10 ⁵ to 10 ¹⁶ CFU/g, 10 ⁵ to 10 ¹⁵ CFU/g, 10 ⁵ to 10 ¹⁴ CFU/g, 10 ⁵ to 10 ¹³ CFU/g , 10 ⁵ to 10 ¹² CFU/g, 10 ⁶ to 10 ¹³ CFU/g, 10 ⁶ to 10 ¹² CFU/g, 10 ⁷ to 10 ¹³ CFU/g, 10 ⁷ to 10 ¹² CFU/g, 10 ⁸ to 10 It may be included in an amount of ¹³ CFU/g or 10 ⁸ to 10 ¹² CFU/g, or may be included in the composition as a culture of live or dead cells in an equivalent number. Specifically, for adult patients, 1X10 ³ to 1X10 ¹⁶ CFU/g of live or dead cells may be administered once or in divided doses. However, the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and those skilled in the art will Taking these factors into consideration, the dosage can be adjusted appropriately. According to one embodiment, the composition includes killed dried strains, and may be administered in an amount of 1g to 10g, 0.5g to 1.5g, 2.5g to 3.5g, or 4.5g to 5.5g at a time.

In one aspect, the pharmaceutical composition may be administered once a day or may be administered several times. Specifically, it may be administered in cycles of 6 days of administration for 7 days followed by 1 day of rest, 5 days of administration followed by 2 days of rest, or 4 days of administration followed by 3 days of rest. More specifically, it may be administered in cycles of 5 days of administration followed by 2 days of rest. there is.

In addition, the pharmaceutically effective amount and effective dosage of the pharmaceutical composition may vary depending on the formulation method, administration method, administration time, and/or administration route of the pharmaceutical composition. In addition, the type and degree of response to be achieved by administration of the pharmaceutical composition, the type of subject to be administered, age, weight, general health condition, symptoms or degree of disease, gender, diet, excretion, simultaneous or It may vary depending on various factors including drugs used together and components of other compositions, and similar factors well known in the pharmaceutical field. A person skilled in the art can easily determine and prescribe an effective dosage for the desired treatment. The pharmaceutical composition according to the present invention can be administered once a day or divided into several times. Therefore, the above dosage does not limit the scope of the present invention in any way. The dosage of the pharmaceutical composition may be 1 ug/kg/day to 1,OOO mg/kg/day.

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 treatment for a metabolic disease or side effects caused by taking a metabolic regulator.

In one embodiment, the composition may be a health functional food composition.

In the health functional food, the “strain”, “first active substance”, “metabolism regulator”, “second active substance”, “co-administration”, etc. may be within the above-mentioned range.

In one embodiment, the health functional food may further include a foodologically acceptable carrier.

As used herein, the term “foodologically acceptable” means that the compound exhibits non-toxic properties to cells or humans exposed to the compound.

In the health functional food, the active ingredient can be added directly to the food or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention or improvement). In general, when manufacturing a food or beverage, the health functional food may be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range.

The health functional food may be formulated with one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, further including one or more of carriers, diluents, excipients, and additives. Foods to which compounds according to one aspect can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, etc.

In addition to containing the effective ingredients, the health functional food may contain other ingredients as essential ingredients without any particular restrictions. For example, like regular beverages, it may contain various flavoring agents or natural carbohydrates as additional ingredients. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. You can. The ratio of the natural carbohydrates can be appropriately determined by the selection of a person skilled in the art.

In addition to the above, health functional foods according to one aspect include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and thickening agents (cheese, chocolate, etc.), pectic acid and salts thereof. , alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These components can be used independently or in combination, and the proportions of these additives can also be appropriately selected by those skilled in the art.

The health functional food may be provided in combination with health functional foods for preventing or improving metabolic diseases known in the art or other existing health functional foods, and the health functional foods for preventing or improving other metabolic diseases are known in the art. It may be a health functional food for preventing or improving metabolic diseases, an existing health functional food, or a newly developed health functional food.

When the health functional food contains other health functional foods that have the effect of preventing or improving metabolic diseases, it is important to mix the amount to obtain the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art. You can.

According to one aspect, the Lactobacillus fermentum strain has anti-obesity activity and the activity of alleviating the side effects of metabolic regulators, and in particular, according to a composition or health functional food for treating metabolic diseases administered in combination with the Lactobacillus fermentum strain and a metabolic regulator. It can be useful for preventing or treating metabolic diseases.

Figure 1 is a graph analyzing the weight gain inhibition effect of strain GB102 according to one embodiment.

Figure 2 is a graph analyzing the effect of improving glucose resistance of strain GB102 according to one embodiment.

Figure 3 is a graph analyzing the effect of improving insulin resistance of strain GB102 according to one embodiment.

Figure 4 is a graph analyzing the effect of increasing energy consumption of the GB102 strain according to one embodiment.

Figure 5 is a graph analyzing succinic acid as a metabolite in a mouse model administered with GB102 strain and other strains according to one embodiment, respectively; MRS: culture medium, DNC01: Lactobacillus fermentum DNCO1 strain

Figure 6 is a graph showing the amount of increase in body weight after single and combined administration of GB102 and dulaglutide in a mouse model with induced obesity.

Figure 7 is a graph showing the results of a glucose tolerance test after single and combined administration of GB102 and dulaglutide in a mouse model with induced obesity.

Figure 8 is a graph analyzed by measuring the weight of brown fat, white fat, liver, and cecum after single and combined administration of GB102 and dulaglutide in an obesity-induced mouse model.

Figure 9 is a graph analyzed by measuring liver levels of AST and ALT after single and combined administration of GB102 and dulaglutide in a mouse model with induced obesity.

Figure 10 is a graph analyzing the levels of triglycerides, high-density cholesterol, low-density cholesterol, and total cholesterol after single and combined administration of GB102 and dulaglutide in a mouse model with induced obesity.

Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the following examples. The embodiments may be subject to various changes, and the embodiments are not limited to the embodiments disclosed below and may be implemented in various forms.

Example 1. Lactobacillus Fermentum ( Lactobacillus fermentum ) Isolation and identification of strain GB102

Isolation and identification of Lactobacillus fermentum GB102 strain was performed by the method described in Korean Patent Application No. 10-2020-0186685 and Korean Patent Application No. 10-2022-0080567. The above document is incorporated herein by reference in its entirety.

Briefly, Lactobacillus fermentum GB102 was isolated from a vaginal sample of a healthy woman who visited the hospital for health checkup. First, vaginal samples were collected with a swab, inoculated into Rogosa SL (MRS) plate medium, and cultured in an anaerobic chamber at 37°C for 48 hours. When bacterial colonies grew, single colonies were subcultured onto new MRS plate medium for pure isolation. After pure isolation, the strain was cultured using MRS medium. Next, among the cultured strains, Lactobacillus fermentum has an inhibitory effect on fat cell accumulation and has low cytotoxicity. Strain GB102 was finally selected. To identify the final selected Lactobacillus fermentum GB102 strain, the 16S rRNA gene of the strain was amplified using primers designed to amplify 16S rRNA and a PCR method, and the corresponding gene was identified using the Sanger sequencing method from the amplified product. The base sequence was secured.

Lactobacillus Fermentum The 16s rRNA sequence of GB102 is shown as SEQ ID NO: 1. The present inventors named the GB102 strain as “ Lactobacillus fermentum GB102” (Accession number: KCTC 14105BP) and transferred it to the Korean collection for type cultures (KCTC) at the Korea Research Institute of Bioscience and Biotechnology in January 2020. Deposited on the 14th. In addition, Lactobacillus fermentum was changed to Limosilactobacillus fermentum . In the following examples, the changed strain names of existing strains are described interchangeably.

Experimental Example 1. Anti-obesity activity following single administration of Lactobacillus Fermentum GB102

To confirm the preventive effect of Lactobacillus fermentum GB102 on metabolic diseases, specifically obesity, GB102 was administered to mice with induced obesity and the inhibition of weight gain, blood sugar control, and energy consumption were analyzed. Specifically, C57BL/6, male mice were fed a normal chow diet (NCD) or a 60% high fat diet (HFD) for 10 weeks, and the normal diet group (G1) and the high-fat diet group (G2) were PBS and GB102 lactic acid bacteria ( ^5x109 cfu/hd) were orally administered at intervals of 2 to 3 days to the high-fat diet + GB102 administration group (G5).

1.1. weight gain inhibition effect

To confirm the effect of suppressing weight gain following the administration of Lactobacillus fermentum GB102, the body weight of the normal diet group (G1), the high-fat diet group (G2), and the high-fat diet + GB102 administration group (G5) were measured every week for 10 weeks.

At this time, the amount of weight gain compared to the initial (0 dya) weight was analyzed for each group. The results are shown in Table 1 and Figure 1 below.

AVG(g)		0 day	1W	2W	3W	4W	5W	6W	7W	8W	9W	10W
G1
	Mean	0	1.10	2.12	3.93	4.77	5.43	5.85	6.20	6.62	7.05	7.98
SEM	0	0.29	0.37	0.24	0.37	0.46	0.45	0.42	0.45	0.38	0.65
G2		Mean	0	2.42	3.58	5.42	8.10	10.83	13.43	15.33	17.87	20.00	21.57
	SEM	0	0.20	0.56	0.54	0.67	0.96	1.05	1.19	1.23	1.13	1.13
G5		Mean	0	1.38	2.48	4.14	6.06	7.62	8.18	9.64	11.10	13.20	14.20
	SEM	0	0.48	0.47	0.58	0.68	0.69	0.73	0.84	1.05	1.11	1.36

Table 1 is a table showing the amount of weight gain in each normal diet group (G1), high-fat diet group (G2), and high-fat diet + GB102 group (G5); G1: Normal diet group (NCD), G2: High-fat diet group (HFD), G5: HFD + GB102

1.2. Blood sugar control effect

To confirm the blood sugar control ability of Lactobacillus Fermentum GB102, an intraperitoneal glucose tolerance test and insulin resistance test were performed after 10 weeks of HFD supply and administration of GB102.

Specifically, for a glucose tolerance test, feed was removed 16 hours before the experiment, the animals were fasted, and only water was supplied. Before intraperitoneally administering 10% glucose (Sigma, 1 g/kg/10mL) solution, fasting blood sugar was measured from the tail vein using a blood glucose measuring device (Aprilis, Auto-check-plus) and disposable blood glucose test strips (Aprilis, Auto-check). Blood sugar was measured again in the tail vein at 30, 60, 90, and 120 minutes after intraperitoneal administration of the glucose solution, and the results are shown in Figure 2.

Specifically, for an insulin tolerance test, mice were fasted for 4.5 hours and then intraperitoneally administered an insulin solution at a dose of 1 U/kg, and after 0 minutes, 30 minutes, 60 minutes, 90 minutes and 120 minutes, the mouse tail was injected. Blood was collected from a vein and blood sugar was measured using a blood glucose meter, and the results are shown in Figure 3.

As shown in Figures 1 to 3, the administration of the GB102 strain to the high-fat diet group (G2) was confirmed to have an effect on suppressing weight gain and improving blood sugar control in mice.

1.3. Effect of increased energy consumption

In order to determine changes in energy consumption following the administration of Lactobacillus Fermentum GB102, oxygen consumption, carbon dioxide generation, and energy consumption were measured through CLAMS (Comprehensive Lab Animal Monitoring System) after 10 weeks of HFD supply and administration of GB102. did.

Specifically, mice from the high-fat diet group (G2) and the high-fat diet + GB102 administration group (G5) were placed in a metabolic cage (Columbus instruments), acclimatized for 24 hours, and metabolic phenomena were observed for 48 hours. Oxygen consumption and energy metabolism were measured using equipment sensors. (Energy expenditure) was measured. The results are shown in Figure 4.

As shown in Figure 4, oxygen consumption, carbon dioxide generation, and energy metabolism significantly increased in mice in the high-fat diet + GB102 administration group (G5) compared to mice in the high-fat diet group (G2). Through this, it was confirmed that energy metabolism (or basal metabolic rate) increased with GB102 administration.

Through the above results, anti-obesity activity following administration of the GB102 strain was confirmed.

Experimental Example 2. Analysis of metabolite changes following administration of Lactobacillus Fermentum GB102

In order to confirm the active ingredient of anti-obesity activity according to the administration of Lactobacillus Fermentum GB102, changes in metabolites according to the administration of GB102 were analyzed.

Specifically, GB102 and L.fermentum DNC01 lactic acid bacteria were cultured in MRS medium for about 16 hours, and then the culture supernatant was separated. 800 μL of methanol was added to 200 μL of the separated culture supernatant, and metabolite extraction was performed through mixer mill (10 minutes) and sonication (10 minutes). Afterwards, the supernatant was filtered and dried through speed vacuum, and the dried sample was re-dissolved in 80% MeOH to a concentration of 20,000 ppm, then placed in 1.5 mL tubes at 100 μL each and dried again. 50 μL of methoxyamine hydrochloride was added to the dried 1.5 mL tube, placed in a thermomixer, and reaction was performed at 30°C for 90 minutes. Afterwards, 50 μL of MSTFA was added, vortexed, placed in a thermomixer, and reaction was carried out at 30°C for 90 minutes. After this, GC-MS analysis was performed, and the results are shown in Figure 5.

As shown in Figure 5, it was confirmed that the detected amount of succinic acid increased with oral administration of Lactobacillus fermentum GB102.

Experimental Example 3. Efficacy of obesity treatment according to combined administration of dulaglutide and Lactobacillus Fermentum GB102

B6 mice (C57BL/6N, male) were used as the animal experiment model. After acclimating 70 mice aged 4 weeks for 2 weeks, they were fed a 60% high fat diet (HFD) for 5-6 weeks starting at 6 weeks of age to induce obesity so that they weighed more than 35 g, as shown in Table 2 below. As shown, 10 or 20 mice were randomly assigned to each group. To confirm the efficacy of combined administration of dulaglutide and GB102 as an obesity treatment, measurements of body weight, blood lipid and liver levels of experimental animals, and analysis of blood sugar control ability were conducted. Additionally, we considered observing animal adverse reactions during administration and necropsy, such as the mouse's reaction to medication and the shine of its fur, and minimized stress, such as minimizing observation points and group caging.

	experimental group	Remark
G1(N=10)	Control(NCD)	PBS 200 ㎕/hd + PBS 150 ㎕/hd
G2(N=20)	Control(HFD)	PBS 200 ㎕/hd + PBS 150 ㎕/hd
G3(N=10)	HFD + Dulaglutide		PBS 200 μl/hd + 1 nmol/kg
G4(N=10)	HFD + Dulaglutide + GB102	5 x 109 cfu/hd + 1 nmol/kg

Table 2 shows the effectiveness of dulaglutide alone or combined administration of dulaglutide and Lactobacillus Fermentum GB102 as an obesity treatment in mice induced with obesity by a high-fat diet according to an embodiment. The total experimental group was divided into four. This is a table showing this.

3.1. Analysis of weight gain inhibition ability

To confirm the weight loss effect of combined administration of dulaglutide and Lactobacillus fermentum GB102, body weight was measured every week during the 6-week administration of dulaglutide and/or GB102.

Specifically, the normal diet group (G1) and the high-fat diet group (G2) were administered PBS (150 ㎕/hd), and the dulaglutide-only group (G3) and the dulaglutide + GB102 combination group (G4) were administered dulaglutide (Eli). Lilly, 1 nmol/kg) was administered subcutaneously once every two days for 4 weeks. At the same time, the dulaglutide + GB102 combination group (G4) was orally administered GB102 lactic acid bacteria (Mediogen, 5x10 ⁹ cfu/head) every day for 6 weeks, and the remaining groups were administered PBS (200 ㎕/hd).

The change compared to the initial body weight was calculated for each group, and the results are shown in Table 3 and Figure 6 below. At this time, a negative value represents a decreased amount of weight change, and a positive value represents an increased amount of weight change.

AVG(g)		0 days	1W	2W	3W	4W	5W	6W
G1
	Mean	0	0.571	0.861	1.526	1.736	2.237	2.504
SEM	0	0.109	0.141	0.224	0.181	0.197	0.201
G2		Mean	0	1.744	4.172	6.588	8.414	10.057	11.142
	SEM	0	0.236	0.254	0.348	0.396	0.442	0.493
G3	Mean	0	-1.544	0.914	2.215	3.437	5.89	7.902
	SEM	0	0.304	0.493	0.846	1.001	0.924	0.863
G4	Mean	0	-2.042	-0.869	-0.544	0.624	3.372	5.172
	SEM	0	0.265	0.411	0.557	0.470	0.402	0.418

Table 3 is a table showing the change in body weight of mice upon co-administration of dulaglutide and Lactobacillus fermentum GB102; G1: Normal diet group (NCD), G2: High-fat diet group (HFD), G3: HFD + Dulaglutide, G4: HFD + Dulaglutide + GB102

As shown in Figure 6 and Table 3, when Lactobacillus Fermentum GB102 and dulaglutide were administered together, body weight was steadily reduced for 6 weeks, so compared to the dulaglutide administration group alone (G3), the combined administration group (G4) A greater weight loss effect was confirmed.

3.2. Analysis of blood sugar control ability

To confirm the blood sugar control ability of combined administration of dulaglutide and Lactobacillus Fermentum GB102, an intraperitoneal glucose tolerance test was performed.

Specifically, an intraperitoneal glucose tolerance test was performed after administration of dulaglutide and/or GB102 for 6 weeks. 16 hours before the experiment, food was removed, the animals were fasted, and only water was provided. Before intraperitoneally administering 10% glucose (Sigma, 1 g/kg/10mL) solution, fasting blood sugar was measured from the tail vein using a blood glucose measuring device (Aprilis, Auto-check-plus) and disposable blood glucose test strips (Aprilis, Auto-check). Blood sugar was measured again in the tail vein at 30, 60, 90, and 120 minutes after intraperitoneal administration of the glucose solution, and the results are shown in Figure 7.

As shown in Figure 7, in the dulaglutide and GB102 combined administration group (G4), blood sugar level significantly decreased 60 minutes after intraperitoneal administration of the glucose solution, so compared to the dulaglutide single administration group (G3), the combined administration group (G4) It was confirmed that the blood sugar control effect was more excellent.

3.3. Weighing brown and white fat, liver, and cecum

To determine changes in organ weight following combined administration of dulaglutide and Lactobacillus fermentum GB102, the weight of fat, liver, and cecum, which are organs related to metabolism, were measured.

Specifically, after 6 weeks of administration of dulaglutide and/or GB102, an autopsy was performed on each subject and brown fat, white fat, liver, and cecum were separated. Afterwards, the weight of each tissue was measured using an electronic scale, and the results are shown in Figure 8.

As shown in Figure 8, a decrease in the weight of organs related to metabolism was confirmed in the dulaglutide single administration group (G3) and the dulaglutide and GB102 combination administration group (G4), and particularly significant changes were confirmed in brown fat and liver.

3.4. Measurement of liver and blood triglyceride and cholesterol levels

Liver levels and blood fat levels were measured following combined administration of dulaglutide and Lactobacillus fermentum GB102.

Specifically, after the administration of dulaglutide and/or GB102 for 6 weeks, blood was obtained from each individual, solidified at room temperature for more than 10 minutes, and then used in a table centrifuge (Eppendorf) at 12000 rpm for 10 minutes to obtain the blood in the blood. Serum was separated. The separated serum was analyzed for aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and total cholesterol (TCHO) using blood biochemical analysis equipment (Fuji, NX700) and a dedicated analysis kit. and high-density cholesterol (HDL) levels were measured. The low-density cholesterol (LDL) value was calculated using the formula below, and the analysis results of each value are shown in Figures 9 and 10.

[Equation 1]

LDL = [TCHO - (HDL + TG/5)]

As shown in Figures 9 and 10, alanine aminotransferase (ALT), triglyceride (TG), and low-density cholesterol (LDL) levels in the dulaglutide single administration group (G3) and the dulaglutide and GB102 combination administration group (G4) A decrease was confirmed.

Through the above results, it was confirmed that the anti-obesity activity and blood sugar control ability were improved by administering Lactobacillus Fermentum GB102 strain in combination with a metabolic regulator, and the prevention or treatment of metabolic diseases was confirmed by co-administering Lactobacillus Fermentum GB102 strain and a metabolic regulator. This means that it can be useful as a composition to prevent the side effects of metabolic diseases.

The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive.

[Accession number]

Name of depository institution: Korea Research Institute of Bioscience and Biotechnology

Accession number: KCTC14105BP

Trust date: 20200114

Claims (35)

1. It contains as an active ingredient a first active substance containing a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator; or

   A pharmaceutical composition for preventing or treating metabolic diseases comprising the first active substance as an active ingredient and the second active substance being administered in combination.
2. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the strain has at least 98.5% sequence identity with the 16S rRNA sequence of SEQ ID NO: 1.
3. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the strain is a strain deposited under accession number KCTC14105BP.
4. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the strain contains a mutation of a naturally occurring Lactobacillus fermentum strain.
5. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the first active substance contains succinic acid.
6. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the first active substance has anti-obesity activity.
7. The method according to claim 6, wherein the anti-obesity activity includes one or more of the following: a pharmaceutical composition for preventing or treating metabolic diseases;

   - Suppresses weight gain;

   - Improvement of glucose or insulin resistance;

   - Increase in basal metabolic rate;

   - Accumulation of brown fat;

   - Inhibition of liver steatosis;

   - Reduction in blood AST (alanine aminotransferase), ALT (alanine transaminase, GPT), triglycerides, low-density cholesterol, high-density cholesterol, or total cholesterol; and

   - Increases the expression of UCP-1 (Uncoupling protein 1).
8. The method of claim 1, wherein the metabolism regulator is a GLP-1 (Glucagon-like peptide-1) receptor agonist, GIP (Glucose-dependent insulinotropic polypeptide) receptor agonist, glucagon agonist, GLP-1/GIP dual receptor agonist, GLP-1/ A pharmaceutical composition for preventing or treating metabolic diseases, wherein one or more types are selected from the group consisting of glucagon dual receptor agonists and GLP-1/GIP/glucagon triple receptor agonists.
9. The method of claim 1, wherein the metabolism regulator is Exenatide, Lixisenatide, Liraglutide, Albiglutide, Taspoglutide, Semaglutide ( Semaglutide, Dulaglutide, Tirzepatide, Cotadutide, Pemvidutide, Efinopegdutide, efocipegtrutide, and A pharmaceutical composition for preventing or treating metabolic diseases selected from the group consisting of Retatrutide.
10. The method of claim 1, wherein the metabolic disease is 1 selected from the group consisting of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, impaired fasting blood glucose, dyslipidemia, lipid metabolism disorder, obesity, fatty liver, insulin resistance syndrome, and glucose tolerance syndrome. A pharmaceutical composition for preventing or treating more than one type of metabolic disease.
11. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the first active material and the second active material are administered in combination simultaneously, sequentially, or in reverse order.
12. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 1, wherein the first and second active substances are administered orally, intravenously, or subcutaneously.
13. The metabolic disease according to claim 1, comprising a first oral preparation containing the first active substance, and a second oral preparation containing the second active substance, wherein the first and second oral preparations are administered orally. Pharmaceutical composition for the prevention or treatment of.
14. A pharmaceutical composition for preventing or treating symptoms caused by side effects of a metabolic regulator using a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof as an active ingredient.
15. The method according to claim 14, wherein the side effects of the metabolism regulator include: a pharmaceutical composition for preventing or treating symptoms caused by side effects of the metabolism regulator;

    - Weight gain after stopping administration of metabolic regulators

    - Increase in blood pressure after stopping administration of metabolic regulators.

    - Deterioration of glycemic control after discontinuation of metabolic regulators; and

    - Increase in glycated hemoglobin after stopping administration of metabolic regulators.
16. The method of claim 14, wherein the metabolism regulator is a GLP-1 (Glucagon-like peptide-1) receptor agonist, GIP (Glucose-dependent insuliontropic polypeptide) receptor agonist, glucagon agonist, GLP-1/GIP dual receptor agonist, GLP-1/ A pharmaceutical composition for preventing or treating symptoms caused by side effects of a metabolic regulator, which is at least one selected from the group consisting of glucagon dual receptor agonists and GLP-1/GIP/glucagon triple receptor agonists.
17. The method of claim 14, wherein the metabolism regulator is Exenatide, Lixisenatide, Liraglutide, Albiglutide, Taspoglutide, Semaglutide ( Semaglutide, Dulaglutide, tirzepatide, cotadutide, Pemvidutide, Efinopegdutide, efocipegtrutide, and A pharmaceutical composition for preventing or treating symptoms caused by side effects of a metabolic regulator selected from the group consisting of Retatrutide.
18. It contains as an active ingredient a first active substance containing a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator; or

    A health functional food for preventing or improving metabolic diseases comprising the first active substance as an active ingredient and the second active substance being administered in combination.
19. The health functional food for preventing or improving metabolic diseases according to claim 18, wherein the first active substance contains succinic acid.
20. The health functional food for preventing or improving metabolic diseases according to claim 18, wherein the first active substance has anti-obesity activity.
21. The method according to claim 20, wherein the anti-obesity activity includes one or more of the following: a health functional food for preventing or improving metabolic diseases;

    - Suppresses weight gain;

    - Improvement of glucose or insulin resistance;

    - Increase in basal metabolic rate;

    - Accumulation of brown fat;

    - Inhibition of liver steatosis;

    - Reduction in blood AST (alanine aminotransferase), ALT (alanine transaminase, GPT), triglycerides, low-density cholesterol, high-density cholesterol, or total cholesterol; and

    - Increases the expression of UCP-1 (Uncoupling protein 1).
22. The method of claim 18, wherein the metabolism regulator is a GLP-1 (Glucagon-like peptide-1) receptor agonist, GIP (Glucose-dependent insuliontropic polypeptide) receptor agonist, glucagon agonist, GLP-1/GIP dual receptor agonist, GLP-1/ A health functional food for preventing or improving metabolic diseases, which is one or more selected from the group consisting of glucagon dual receptor agonists and GLP-1/GIP/glucagon triple receptor agonists.
23. The method of claim 18, wherein the metabolism regulator is Exenatide, Lixisenatide, Liraglutide, Albiglutide, Taspoglutide, Semaglutide ( Semaglutide, Dulaglutide, tirzepatide, cotadutide, Pemvidutide, Efinopegdutide, efocipegtrutide, and A health functional food for preventing or improving metabolic diseases selected from the group consisting of Retatrutide.
24. The method of claim 18, wherein the metabolic disease is 1 selected from the group consisting of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, impaired fasting glucose, dyslipidemia, lipid metabolism disorder, obesity, fatty liver, insulin resistance syndrome, and glucose intolerance syndrome. Health functional food for preventing or improving metabolic diseases of more than one type.
25. The health functional food for preventing or improving metabolic diseases according to claim 18, wherein the first active substance and the second active substance are administered in combination simultaneously, sequentially, or in reverse order.
26. The method of claim 18, wherein the metabolic disease comprises a first oral formulation containing the first active material and a second oral formulation containing the second active material, and the first and second oral formulations are administered orally. Health functional food for the prevention or improvement of.
27. A health functional food for preventing or improving symptoms caused by side effects of metabolic regulators using Lactobacillus fermentum strains, cultures of the strains, lysates of the strains, or mixtures thereof as active ingredients.
28. The method of claim 27, wherein the side effects of the metabolism regulator include: a health functional food for preventing or improving symptoms caused by side effects of the metabolism regulator;

    - Weight gain after stopping administration of metabolic regulators

    - Increase in blood pressure after stopping administration of metabolic regulators.

    - Deterioration of glycemic control after discontinuation of metabolic regulators; and

    - Increase in glycated hemoglobin after stopping administration of metabolic regulators.
29. The method of claim 27, wherein the metabolism regulator is Exenatide, Lixisenatide, Liraglutide, Albiglutide, Taspoglutide, Semaglutide ( Semaglutide, Dulaglutide, tirzepatide, cotadutide, Pemvidutide, Efinopegdutide, efocipegtrutide, and A health functional food for preventing or improving symptoms caused by side effects of a metabolic regulator selected from the group consisting of Retatrutide.
30. A first active substance comprising a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator; or

    A method for preventing or treating metabolic disease, comprising administering to an individual in need a composition comprising the first active material and administering the second active material in combination.
31. A method for preventing or treating symptoms caused by side effects of a metabolic regulator, comprising administering a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof to an individual in need thereof.
32. It contains as an active ingredient a first active substance containing a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator; or

    Use for the manufacture of a composition for preventing or treating metabolic diseases comprising the first active substance as an active ingredient and co-administering the second active substance.
33. It contains as an active ingredient a first active substance containing a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof, and a second active substance containing a metabolic regulator; or

    Use for the manufacture of a health functional food composition for preventing or improving metabolic diseases, which contains the first active substance as an active ingredient and where the second active substance is administered in combination.
34. Use of a Lactobacillus fermentum strain, a culture of the strain, a lysate of the strain, or a mixture thereof in the manufacture of a composition for preventing or treating symptoms caused by side effects of metabolic regulators.
35. Use of Lactobacillus fermentum strains, cultures of the strains, lysates of the strains, or mixtures thereof in the manufacture of health functional food compositions for preventing or improving symptoms caused by side effects of metabolic regulators.

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