A composition for improving obesity comprising an active ingredient derived from lactobacillus plantarum belp1

Abstract

The present invention relates to a composition comprising the Lactobacillus plantarum beLP1 strain as an active ingredient, and provides a food and pharmaceutical composition for the prevention and treatment of obesity-related diseases.

Description

Technology Field

[0001] The present invention relates to Lactobacillus plantarum ( Lactobacillus plantarum The present invention relates to a composition containing a beLP1 strain or a component derived therefrom, specifically to a food or pharmaceutical composition capable of preventing or improving symptoms associated with obesity and metabolic diseases. Background Technology

[0002] Obesity is a chronic metabolic disease caused by the excessive accumulation of fat in the body due to an imbalance between energy intake and expenditure, and it acts as a major risk factor for various complications such as type 2 diabetes, hypertension, dyslipidemia, cardiovascular disease, and fatty liver.

[0003] Recently, the prevalence of obesity has been rapidly rising worldwide due to the increase in Westernized dietary habits and sedentary lifestyles, and consequently, the demand for functional foods and pharmaceutical materials that can prevent and improve obesity is continuously increasing.

[0004] In particular, as the link between gut microbes and metabolic diseases has been proven through various studies, an approach to improving obesity through the regulation of the gut environment is emerging as a new alternative.

[0005] Obese patients generally have low gut microbial diversity and an imbalance between Firmicutes and Bacteroidota, which negatively affects metabolism, such as increasing energy extraction efficiency and fat storage.

[0006] Accordingly, numerous bioactive compositions based on gut microorganisms, such as probiotics, prebiotics, synbiotics, and postbiotics, are being developed. Among these, probiotics have been reported to have preventive effects against various diseases, including obesity, by directly supplying beneficial bacteria into the body to improve the gut flora and suppress inflammation.

[0007] Representative anti-obesity related probiotic strains include Lactobacillus rhamnosus , Bifidobacterium breve , Lactobacillus gasseri These include, and have demonstrated effects such as weight loss, reduction of visceral fat, and increased insulin sensitivity in animal or human clinical trials.

[0008] However, existing live bacteria-based probiotics technology regulates the intestinal environment and improves the body's immune and metabolic functions by directly consuming beneficial microorganisms; as such live bacteria-based technology, it inherently entails several technical limitations.

[0009] In other words, live bacteria are highly sensitive to stomach acid and bile acids, making it difficult for them to reach the intestines alive. Furthermore, their activity is prone to loss during storage and distribution, and consistent physiological activity is difficult to ensure due to significant variations in efficacy depending on an individual's intestinal environment and immune status.

[0010] Furthermore, as the risk of infection is raised in high-risk groups with weakened immunity, there are structural limitations to the direct use of live bacteria in terms of safety and stability.

[0011] Meanwhile, numerous chemical anti-obesity materials based on specific natural extracts or synthetic compounds have been proposed in the past; for example, caffeine, catechin, Garcinia extract (HCA), and orlistat have claimed efficacy in breaking down body fat, suppressing appetite, and inhibiting fat absorption.

[0012] However, these compound-based anti-obesity technologies often have efficacy limited to a single mechanism, and side effects such as liver toxicity, gastrointestinal disorders, reduced absorption, and rebound weight gain have been reported with long-term use. Furthermore, some have limitations, such as a lack of clinical significance or minimal metabolic improvement effects. Additionally, these compound-based anti-obesity agents are insufficient to implement complex physiological mechanisms, such as improving the gut environment, suppressing inflammation, and regulating immunity.

[0013] Accordingly, the concept of postbiotics, which utilizes dead cells or strain-derived substances rather than live bacteria, has recently emerged as an alternative.

[0014] Postbiotics are compositions containing metabolites, cell wall fragments, dead cells, etc., produced by beneficial bacteria. They have the advantage of resolving the instability issues of live bacteria while exhibiting similar effects in promoting the proliferation of beneficial intestinal bacteria, maintaining immune balance, and providing anti-inflammatory benefits.

[0015] In particular, dead cell-based compositions have high storage stability, are free from the risk of infection, and can clearly identify the physiological activity of specific components, making them highly applicable as functional food and pharmaceutical compositions.

[0016] However, most postbiotic-based anti-obesity research to date has either applied dead cells of specific lactic acid bacteria alone or has been limited to immune-modulating effects, resulting in a lack of development in postbiotic compositions that encompass complex mechanisms such as obesity prevention and liver function improvement.

[0017] Under these circumstances, the inventors completed the present invention by identifying that a newly isolated Lactobacillus plantarum strain can exhibit anti-obesity effects through various complex mechanisms, and intended to provide a postbiotic-based functional composition using the same. The problem to be solved

[0018] The present invention aims to solve the problems of the aforementioned prior art, and the objective of the present invention is to provide a composite functional composition capable of simultaneously improving various physiological functions, including anti-obesity effects. means of solving the problem

[0019] According to one aspect of the present invention, Lactobacillus plantarum ( Lactobacillus plantarumA pharmaceutical composition for the prevention or treatment of obesity-related diseases is provided, comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0020] In one embodiment, the obesity-related disease may be selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome.

[0021] According to another aspect of the present invention, Lactobacillus plantarum ( Lactobacillus plantarum A food composition for improving obesity-related diseases is provided, comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0022] According to another aspect of the present invention, Lactobacillus plantarum ( Lactobacillus plantarum An anti-obesity food composition is provided comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0023] According to another aspect of the present invention, Lactobacillus plantarum ( Lactobacillus plantarum A food composition for weight loss and liver function improvement is provided, comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0024] In one embodiment, the food composition can inhibit liver fat accumulation. Effects of the invention

[0025] A composition according to one aspect of the present invention is Lactobacillus plantarum ( Lactobacillus plantarum By including the beLP1 strain or a component derived therefrom, it is possible to suppress body weight gain induced by a high-fat diet and demonstrate effects of improving liver function indicators (AST, ALT) and reducing adipose tissue weight.

[0026] In addition, the above composition can be utilized as a multifunctional material that can effectively contribute to the prevention or improvement of obesity and related metabolic diseases.

[0027] The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention. Brief explanation of the drawing

[0028] Figure 1 shows the results of measuring body weight (A) and body weight change rate (B) following the administration of the beLP1 strain. Figure 2 shows the differences in AST (A), ALT (B), and triglycerides (C) measurements following the administration of the beLP1 strain. Figure 3 shows the difference in liver and fat weight measurements following the administration of the beLP1 strain. Specific details for implementing the invention

[0029] The terms used in this specification have been selected to be as widely used as possible, taking into account their functions in the present invention; however, they may vary depending on the intent of those skilled in the art, case law, the emergence of new technologies, etc.

[0030] In addition, in specific cases, there are terms arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, terms used in this invention must be defined not merely by their names, but based on their meanings and the overall content of the invention.

[0031] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by those skilled in the art to which the present invention pertains. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and should not be interpreted in an ideal or overly formal sense unless explicitly defined in this application.

[0032] Numerical ranges include the values ​​defined in the above ranges. All maximum numerical limits given throughout this specification include all lower numerical limits as clearly written. All minimum numerical limits given throughout this specification include all higher numerical limits as clearly written. All numerical limits given throughout this specification will include all better numerical ranges within a wider numerical range, as clearly written.

[0033] The embodiments of the present invention are described in detail below, but it is obvious that the present invention is not limited by the following embodiments.

[0034] Pharmaceutical composition

[0035] According to one aspect of the present invention, Lactobacillus plantarum ( Lactobacillus plantarum A pharmaceutical composition for the prevention or treatment of obesity-related diseases is provided, comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0036] The present invention relates to a newly isolated Lactobacillus plantarum ( Lactobacillus plantarumBy utilizing the beLP1 strain or various forms of components derived therefrom, a complex functional composition capable of effectively preventing or improving obesity and related metabolic diseases is provided.

[0037] The above “Lactobacillus plantarum ( Lactobacillus plantarum )” is a Gram-positive rod-shaped bacterium belonging to the phylum Firmicutes and the family Lactobacillaceae. It is a facultative anaerobic lactic acid bacterium commonly found in plant-based fermented foods (e.g., kimchi, pickled vegetables, fermented grains, etc.) or in the human intestinal environment.

[0038] The above-mentioned Lactobacillus plantarum has high resistance to gastric acid and bile acid, so it has a high survival rate in the gastrointestinal tract and can stably colonize the intestines. Based on its extensive carbohydrate metabolic ability, it can produce organic acids and antimicrobial substances (e.g., bacteriocins) to inhibit the growth of pathogenic microorganisms and contribute to maintaining the balance of intestinal microorganisms.

[0039] The above-mentioned Lactobacillus plantarum is a beneficial bacterium that is reported to have the function of regulating the expression of inflammatory cytokines and strengthening intestinal mucosal immunity, and is capable of exhibiting various physiological activities such as contributing to energy metabolism and improving the intestinal environment by inducing the production of short-chain fatty acids (SCFAs).

[0040] Meanwhile, the above-mentioned Lactobacillus plantarum beLP1 strain was isolated from "Kimchi" and is a Gram-positive bacillus; analysis of its 16S rDNA sequence revealed 99% homology with Lactobacillus plantarum.

[0041] Accordingly, the inventors deposited the newly isolated Lactobacillus plantarum beLP1 strain with the Korea Research Institute of Bioscience and Biotechnology (KRIBB), a depositary institution under the Treaty of Budapest, on May 7, 2024, under accession number KCTC15902BP.

[0042] The above-mentioned Lactobacillus plantarum beLP1 strain is a microorganism newly isolated from kimchi, based on the safety and physiological benefits of lactic acid bacteria derived from fermented foods, while previously reported L. plantarum It is a strain that is distinguished from other strains in terms of specific functional activity and genetic characteristics.

[0043] It is a well-known fact in the field of microbiology that microorganisms are considered to be the same species when their 16S rDNA similarity is 98.7% to 99% or higher, but that significant functional differences may exist depending on their functional profile, ability to produce metabolites, and characteristics of interaction with the host.

[0044] Based on the genetic identification results and the differences in physiological activity confirmed through experiments, the inventors [described] this strain as previously reported L. plantarum A novel strain with functionalities distinct from strains was named Lactobacillus plantarum beLP1.

[0045] The aforementioned Lactobacillus plantarum beLP1 strain has not only proven safety based on dietary experience in that it is derived from kimchi, but also, existing L. plantarum It was confirmed to possess differentiated functional characteristics that can induce complex physiological activities, such as inhibition of weight gain, improvement of liver function, and reduction of adipose tissue weight, when compared to other classes.

[0046] In particular, the above-mentioned Lactobacillus plantarum beLP1 strain maintains significant physiological activity not only in live cells but also in various forms such as dead cells, lysates, and cultures, thus having excellent potential for industrial utilization as a postbiotic-based functional material.

[0047] Consequently, the inventors of the present invention... L. plantarumBy securing a novel strain beLP1 that is genetically similar to the species but clearly distinguishable functionally, and by providing a functional composition capable of improving obesity-related diseases based on this, we have realized a bioactive technological advancement that is fundamentally differentiated from existing lactic acid bacteria-based technologies.

[0048] Accordingly, the above pharmaceutical composition comprises one or more active ingredients selected from the group consisting of Lactobacillus plantarum beLP1 strain, lysate thereof, culture thereof, concentrate thereof, dried thereof, and dead cells thereof, and can be utilized for the prevention or treatment of obesity-related diseases.

[0049] The term “prevention” above refers to any act that reduces the frequency or severity of pathological phenomena. Prevention may be complete or partial. In this case, it may refer to a phenomenon in which symptoms of obesity-related diseases within an individual are reduced compared to the case where the composition is not used.

[0050] The aforementioned “treatment” refers to any act of clinical intervention intended to alter the natural processes of the target or cell to be treated, and may be performed during the progression of a clinical pathological condition or to prevent it. The intended therapeutic effects may include preventing the onset or recurrence of the disease, alleviating symptoms, reducing all direct or indirect pathological consequences associated with the disease, preventing metastasis, slowing the rate of disease progression, alleviating or temporarily resolving the disease state, or improving the prognosis.

[0051] The aforementioned “obesity-related diseases” is a concept that encompasses various metabolic, cardiovascular, and immune diseases caused by imbalances in energy metabolism and chronic low-grade inflammatory conditions, going beyond the simple accumulation of body fat. Representative examples include fatty liver (non-alcoholic fatty liver disease, NAFLD), type 2 diabetes, hyperlipidemia, dyslipidemia, insulin resistance, hypertension, arteriosclerosis, cardiovascular disease, and metabolic syndrome.

[0052] The aforementioned diseases are closely related to one another, and generally tend to manifest as a complex combination of multiple diseases centered around obesity.

[0053] For example, in obese patients, the excessive accumulation of visceral fat leads to the secretion of inflammatory cytokines from fat cells, which results in fatty liver where fat is abnormally accumulated in liver cells and can cause an increase in liver function indicators (AST, ALT).

[0054] In addition, in obese patients, reduced insulin sensitivity impairs blood glucose control, increasing the risk of type 2 diabetes, and elevated blood triglyceride and LDL cholesterol levels increase the incidence of hyperlipidemia and arteriosclerosis. Furthermore, these conditions can exacerbate chronic hypoinflammatory conditions and lead to cardiovascular complications and systemic metabolic diseases.

[0055] That is, obesity-related diseases are closely linked to chronic low-grade inflammatory states and pathological changes in visceral fat induced by obesity, and the pharmaceutical composition can contribute to preventing the occurrence of or inhibiting the progression of said diseases through various mechanisms of action, such as inhibiting weight gain, improving liver function, regulating blood lipid levels, and restoring the balance of intestinal microorganisms.

[0056] Meanwhile, in the present invention, the term “crushed material” may refer to a product obtained by crushing the Lactobacillus plantarum strain of the present invention by a known method.

[0057] In the present invention, the term "culture" means growing the Lactobacillus plantarum strain of the present invention under appropriately controlled environmental conditions.

[0058] The above culture process can be carried out according to suitable media and culture conditions known in the art, and can be easily adjusted and used by those skilled in the art depending on the selected strain. Specifically, the above culture may be batch, continuous, or fed-batch, but is not limited thereto.

[0059] For example, the culture of the above-mentioned Lactobacillus plantarum strain may be carried out in a nutrient medium (NA medium) under one or more of the following conditions: a temperature of 25°C to 40°C, a stirring speed of 50 rpm to 200 rpm, and a pH of 6 to 9, but is not limited thereto.

[0060] In addition, it can be performed under relative humidity conditions of 40% to 50% or temperature conditions of 30°C to 40°C based on 24-hour relative humidity, but is not limited thereto.

[0061] In the present invention, the term "culture medium" refers to a culture medium obtained by culturing the Lactobacillus plantarum strain, and may be a culture medium containing the strain or a cell-free culture medium from which the cells have been removed.

[0062] The above culture may be a culture medium obtained after culturing the Lactobacillus plantarum beLP1 strain in a liquid culture medium, and specifically, the culture medium of the Lactobacillus plantarum beLP1 strain may be a culture stock containing the cells, and may also be cells obtained by removing or concentrating the culture supernatant.

[0063] The composition of the above culture medium may additionally include components that act synergistically on the growth of Lactobacillus, in addition to the components required for the normal culture of Lactobacillus, and such a composition can be easily selected by a person skilled in the art. Furthermore, the form of the above culture may be liquid or solid, and is not limited to either one.

[0064] In the present invention, the term "concentrate" means a culture of the strain concentrated by a conventional method. The concentrate may be a concentrate obtained by concentrating a liquid culture medium in which the Lactobacillus plantarum beLP1 strain is cultured by a conventional method, but the nature of the culture is not limited to liquid.

[0065] In the present invention, the terms “dry product” or “dry product of culture” refer to a culture of the strain dried by a conventional method. The dried product of culture may be a concentrate obtained by drying a liquid culture medium in which the Lactobacillus plantarum beLP1 strain was cultured by a conventional method, but the nature of the culture is not limited to liquid.

[0066] The above-mentioned Lactobacillus plantarum strain or its dead cells may be any of those prepared by a known method, and they do not exhibit toxicity and are harmless to the human body.

[0067] In addition, the above-mentioned dead cells can be prepared by heat-treating the corresponding live cells or treating them with formalin or other disinfectants, and the dead cells can be used even if they are substantially dead.

[0068] In addition, the above-mentioned dead cell may be a suspension of dead cells or a dried product obtained by washing and centrifuging a strain obtained by culturing by a conventional method, repeating washing and dehydration as needed, suspending it in distilled water, physiological saline, etc., and heating the suspension at, for example, 60 to 140°C for 1 to 60 minutes, or a suspension of dead cells or a dried product obtained by irradiating the above-mentioned dead cell suspension with gamma rays or neutron rays.

[0069] The drying means for the above-mentioned dead cell suspension is not particularly limited as long as it is a known drying means, but examples include spray drying and freeze-drying. In some cases, enzyme treatment, surfactant treatment, or grinding / crushing treatment may be performed before or after sterilization treatment by heating, etc., or before or after drying treatment, and the dead cells obtained by such treatment are also included in the dead cells of the present invention. Furthermore, the above-mentioned dead cells may be manufactured in the following manner, but are not limited thereto:

[0070] The above Lactobacillus plantarum beLP1 strain is a probiotic strain, can be utilized in various forms, is harmless to the human body, and can be used without side effects.

[0071] The above "probiotics" refer to live bacteria that enter the body and have beneficial effects on health. Most probiotics known to date have been consumed as fermented milk products made using lactic acid bacteria such as Lactobacillus, but recently, products containing some strains other than Lactobacillus, such as Bifidobacterium and Enterococcus, are being sold in the form of fermented milk, granules, and powders.

[0072] The above Lactobacillus plantarum beLP1 strain may also be used in the form of fermented milk, granules, powder, etc., but is not limited thereto.

[0073] The Lactobacillus plantarum beLP1 strain of the present invention can be cultured using a conventional culture method for Lactobacillus strains. Natural or synthetic media may be used as the culture medium.

[0074] For example, glucose, sucrose, dextrin, glycerol, starch, etc. may be used as carbon sources for the above medium, and peptone, meat extract, yeast extract, dried yeast, soybeans, ammonium salts, nitrates, and other organic or inorganic nitrogen-containing compounds may be used as nitrogen sources, but are not limited to these components.

[0075] Inorganic salts included in the above medium may include magnesium, manganese, calcium, iron, potassium, etc., but are not limited to these.

[0076] In addition to the components of the carbon source, nitrogen source, and inorganic salt mentioned above, amino acids, vitamins, nucleic acids, and related compounds may be added to the medium, and the strain may be cultured for 12 hours to 4 days at a temperature range of 20 to 40 ℃.

[0077] Specifically, the culture medium of the newly isolated strain may be a culture stock containing the cells, or it may be a culture supernatant from which the supernatant has been removed or concentrated.

[0078] The composition of the above culture medium may additionally include components that act synergistically on the growth of Lactobacillus, in addition to the components required for the cultivation of Lactobacillus in general, and the composition accordingly can be easily selected by a person skilled in the art.

[0079] In addition, the strain may be in a liquid or dry state, and drying methods may include air drying, natural drying, spray drying, and freeze-drying, but are not limited thereto.

[0080] Meanwhile, the above pharmaceutical composition may be administered in the form of oral delivery or parenteral delivery. The composition may be administered systemically or topically, and said administration may include oral administration and parenteral administration. The composition may be formulated with a suitable amount of pharmaceutically acceptable vehicle or carrier to provide an appropriate form of administration.

[0081] The above composition may further include a carrier, an excipient, and a diluent used in the manufacture of a pharmaceutical composition. Examples of the carrier, excipient, and diluent include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil.

[0082] In addition, the above composition can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injectable solutions.

[0083] Solid dosage forms for oral administration may include tablets, pills, powders, granules, capsules, etc., and the solid dosage forms may be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. In addition, in addition to the excipients, lubricants such as magnesium styrate and talc may be used.

[0084] Liquid formulations for oral administration may include suspensions, liquid formulations, emulsions, syrups, etc., and in addition to simple diluents such as water and liquid paraffin, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be used.

[0085] Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. For the non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. Witepsol, macrogol, tween 61, cacao oil, laurin oil, and glycerogelatin may be used as bases for the suppositories.

[0086] The above pharmaceutical composition may be administered to a subject in a pharmaceutically effective amount. The “pharmaceutically effective amount” refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including the type and severity of the patient’s disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0087] The above pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, administered sequentially or simultaneously with conventional therapeutic agents, and administered as a single or multiple doses. It is preferable to administer an amount that obtains maximum effect with a minimum amount without side effects, taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art.

[0088] Food composition

[0089] Another aspect of the present invention is Lactobacillus plantarum ( Lactobacillus plantarum The present invention provides a food composition for improving obesity-related diseases comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0090] The above “food composition” refers to a composition in which the above active ingredient is prepared in the form of a general food or a form having specific functionality, and processed so that it can be consumed by the human body.

[0091] The above food composition can be applied in various formulations such as tablets, capsules, powders, liquids, jellies, and beverages, and can be manufactured into dietary supplements, functional convenience foods, or functional beverages.

[0092] The above Lactobacillus plantarum beLP1 strain is a lactic acid bacteria-derived substance in GRAS (Generally Recognized As Safe) status, and its safety is ensured, so it can be consumed over a long period of time with minimal burden and can be applied in the form of daily food intake.

[0093] In particular, since the above food composition can control risk factors for obesity-related diseases through complex physiological mechanisms such as inhibiting weight gain, regulating fat metabolism, and improving liver function, it can be used as a raw material capable of displaying functional claims such as body fat reduction and improvement of blood lipids according to the functional evaluation standards of health functional foods.

[0094] Accordingly, the above food composition has very high potential for industrial application as a health functional food aiming for pharmaceutical-level physiological activity, going beyond general food.

[0095] Another aspect of the present invention is Lactobacillus plantarum ( Lactobacillus plantarum The present invention provides an anti-obesity food composition comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0096] The aforementioned "anti-obesity" is a concept that includes physiological functions of preventing the onset of obesity or mitigating its progression by inhibiting the excessive accumulation of body fat, inducing a reduction of accumulated fat, and further regulating the balance of energy metabolism.

[0097] The aforementioned anti-obesity effects can be realized through the inhibition of weight gain, reduction of visceral or subcutaneous fat, reduction of fat cell size, improvement of lipid metabolism, or increased insulin sensitivity, and these effects may manifest not only through a single pathway but also through various physiological mechanisms such as inhibition of inflammation and changes in hormonal response.

[0098] The above food composition can exhibit complex anti-obesity effects through the inhibition of weight gain, improvement of liver function, and reduction of fat tissue weight, and can be applied in the form of a general food or a health functional food, and can be utilized as a dietary supplement anti-obesity material that exhibits excellent functionality while being safe for long-term consumption.

[0099] Another aspect of the present invention is Lactobacillus plantarum ( Lactobacillus plantarum The present invention provides a food composition for weight loss and liver function improvement comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells.

[0100] The aforementioned “weight reduction” refers to physiological changes in which total body weight decreases through the promotion of body fat breakdown, inhibition of fat accumulation, or activation of energy metabolism, and this includes cases involving a reduction in the weight of adipose tissue, such as visceral fat and subcutaneous fat.

[0101] The above “improvement of liver function” means a state in which the metabolic activity of liver cells is maintained or restored to normal, blood levels of liver enzymes (AST, ALT, etc.) are stabilized, and fat accumulation and inflammatory responses are reduced.

[0102] The improvement of the aforementioned liver function is closely related to the prevention or alleviation of fatty liver (particularly non-alcoholic fatty liver disease, NAFLD) or metabolic liver disease.

[0103] The above food composition can exhibit complex physiological activity by simultaneously regulating pathophysiological factors that cause liver dysfunction and weight gain.

[0104] In particular, the above food composition can have the effect of preventing the progression of fatty liver or alleviating its severity by inhibiting the abnormal accumulation of triglycerides in liver tissue.

[0105] The above-mentioned liver fat accumulation activity can be achieved by regulating the fatty acid synthesis pathway within the liver.

[0106] Consequently, the above food composition can be usefully utilized as a functional food material capable of simultaneously improving liver health along with weight loss.

[0107] Therefore, the above food composition can be used as a health functional food for alleviating various symptoms associated with obesity.

[0108] The above-mentioned health functional food refers to a food manufactured and processed using raw materials or ingredients having functional properties useful to the human body in accordance with the Health Functional Foods Act, and the above-mentioned functionality may refer to consumption for the purpose of obtaining effects useful for health purposes, such as regulating nutrients or physiological actions regarding the structure and function of the human body.

[0109] The above food composition may include conventional food additives, and unless otherwise specified, the suitability of said food additives may be determined in accordance with the specifications and standards for the relevant items, in accordance with the general provisions and general test methods of the food additive code approved by the Ministry of Food and Drug Safety.

[0110] The items listed in the above food additives code may include, for example, chemically synthesized products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid; natural additives such as persimmon dye, licorice extract, crystalline cellulose, sorghum dye, and guar gum; and mixed preparations such as L-sodium glutamate preparations, alkaline noodle additives, preservative preparations, and tar dye preparations.

[0111] The above food composition can be used in various ways for the improvement of obesity-related symptoms or diseases, such as in foods and beverages, and, for example, can be used in various types of foods, beverages, chewing gum, tea, vitamin complexes, health functional supplements, food additives, etc.

[0112] In addition, the above-mentioned health functional food may be manufactured and processed into any one formulation selected from the group consisting of tablets, granules, powder, capsules, liquid solutions, and pills for the purpose of improving obesity-related symptoms or diseases.

[0113] Specifically, the above-mentioned health functional food in tablet form may be manufactured by granulating a mixture of active ingredients, excipients, binders, disintegrants, and other additives by a conventional method, and then adding a lubricant or the like and compression molding, or by directly compression molding the mixture. In addition, the above-mentioned health functional food in tablet form may contain a sweetener or the like as needed, and may be coated with a suitable coating agent as needed.

[0114] Among the above-mentioned capsule-type health functional foods, hard capsules can be manufactured by filling a conventional hard capsule with a mixture of an active ingredient and additives such as excipients, or the granules thereof, or coated granules, and soft capsules can be manufactured by filling a capsule base such as gelatin with a mixture of an active ingredient and additives such as excipients. The above-mentioned soft capsules may contain plasticizers such as glycerin or sorbitol, coloring agents, preservatives, etc., as needed.

[0115] The above-mentioned pill-shaped health functional food may be prepared by molding a mixture of active ingredients, excipients, binders, disintegrants, etc., in a suitable way, and if necessary, the pill may be coated with sucrose or other suitable coating agents, or coated with starch, talc, or a suitable substance.

[0116] The above granular health functional food can be manufactured into a granular form by a suitable method using a mixture of active ingredients, excipients, binders, disintegrants, etc., and may contain flavoring agents, sweeteners, etc. as needed.

[0117] In addition, the definitions of terms for the above excipients, binders, disintegrants, lubricants, sweeteners, flavorings, etc. may include those described in literature known in the art and having the same or similar functions.

[0118] The present invention will be further described in detail through the following examples, but it is obvious that the present invention is not limited by the following examples.

[0119] Preparation Example 1: Sample Acquisition and Strain Isolation

[0120] Lactobacillus plantarum beLP1 was isolated and identified from "kimchi".

[0121] The kimchi sample was roughly filtered through a 160 μm sieve, and the filtrate was diluted with sterile saline. The obtained sample was serially diluted and inoculated onto MRS solid medium (288210, Difco's Lactobacilli MRS Agar), and the plates were cultured at 37°C for 48 hours.

[0122] A single colony was selected from the plate and plated onto a new MRS agar plate. The plate was incubated at 37°C for 36 hours. Then, each colony was subcultured up to 3 times for bacterial purification.

[0123] Isolated colonies were used for Gram staining, catalytic testing, and genetic information verification via 16S rRNA. The Lactobacillus plantarum beLP1 strain was further cultured in MRS liquid medium (Laboratorios Conda, Spain) and stored in a 20% (v / v) glycerol solution at -80°C until further use.

[0124] To identify the strain more accurately, molecular phylogenetic methods based on DNA sequencing were performed.

[0125] The strain was identified by submitting colonies grown on MRS solid medium (Difco™ Lactobacilli MRS Agar) to MACROGEN for 16S rRNA sequencing analysis.

[0126] The primers used are 785F (5'-GGA TTA GAT ACC CTG GTA-3'), 907R (5'-CCG TCA ATT CMT TTR AGT TT-3'), 27F (5'-AGA GTT TGA TCM TGG CTC AG-3'4) and 1492R (5'-TAC GGY TAC CTT GTT ACG ACT T-3)'.

[0127] As a result of DNA sequencing analysis, it was confirmed that the strain has up to 99% homology with strains of the genus Lactobacillus.

[0128] Using the Megablast program, a BLAST search was performed on the standard type material sequences of the rRNA / ITS database (16S ribosomal RNA sequences) and the 16S rRNA gene sequences of the experimental bacteria.

[0129] Preparation Example 2: Preparation of culture solution of Lactobacillus plantarum beLP1

[0130] Live Lactobacillus plantarum beLP1 strains are pre-cultured by aerobic or anaerobic culturing in media commonly used for lactic acid bacteria, and then cultured for 1 to 3 days while maintaining a pH of 5.0 to 8.0 and a temperature of 20 to 40°C to obtain a dry weight (DW) of 3.0 × 10⁶ 12 This culture was performed until the number of cells reached cfu / g or more.

[0131] Afterwards, the supernatant was separated by centrifugation at 8,000 rpm for 10 minutes. The separated supernatant was diluted 10-fold (10%, v / v) to prepare a culture medium for Lactobacillus plantarum beLP1.

[0132] Preparation Example 3: Preparation of inactivated cells of Lactobacillus plantarum beLP1

[0133] Lactobacillus plantarum beLP1 isolated in Experimental Example 1 was heat-treated at 80 to 115°C for 5 to 30 minutes to inactivate the cells, then separated and recovered using a continuous centrifuge, and subsequently freeze-dried and powdered.

[0134] Experimental Example 1: Evaluation of weight change in a high-fat diet-induced obesity model

[0135] The effects of a component derived from Lactobacillus plantarum beLP1 on body weight change were evaluated in a high-fat diet (HFD)-induced obese mouse model.

[0136] 7-week-old male C57BL / 6N mice were used and, after a one-week acclimatization period, were classified into a normal diet group (Normal, n=5), a high-fat diet group (HFD, n=10), and a high-fat diet + beLP1 administration group (HFD + beLP1, n=10). Obesity (individual body weight 30 g or more) was induced in the normal group by feeding a normal diet, while the HFD and HFD + beLP1 groups were fed a high-fat diet with 60% fat content for 3 weeks. Additionally, beLP1 dead cells (30 billion cells / day) were orally administered to the HFD + beLP1 group daily for 10 weeks.

[0137] Body weight was measured twice a week starting from the start of the experiment (Day 0), and finally, body weight and the rate of change in body weight were compared after 10 weeks.

[0138] Referring to Figure 1, the HFD group showed a significant increase in body weight compared to the normal diet group, but the HFD + beLP1 administration group showed a tendency for weight gain to be suppressed despite the same high-fat diet conditions.

[0139] In particular, when comparing the rate of change in body weight (%), the beLP1 administration group showed significantly lower values ​​compared to the HFD group, confirming a statistically significant effect of inhibiting weight gain (*p < 0.05).

[0140] The above results indicate that beLP1 can effectively inhibit weight gain during the process of obesity induction by a high-fat diet, providing experimental evidence supporting its potential as a food or pharmaceutical composition for the prevention or treatment of obesity.

[0141] Experimental Example 2: Evaluation of Changes in Blood Biochemical Indicators

[0142] Obesity was induced in 7-week-old C57BL / 6N male mice using a high-fat diet (HFD), and blood biochemical indicators were analyzed after oral administration of beLP1 dead cells (30 billion cells / day) for 10 weeks.

[0143] The experimental groups consisted of a normal diet group, a high-fat diet group, and a high-fat diet + beLP1 administration group. At the end of the experiment, blood was collected and serum was separated, and the levels of ALT (alanine aminotransferase), AST (aspartate aminotransferase), and triglycerides (TG) were measured using an automated biochemical analyzer.

[0144] Referring to Figure 2, the HFD group showed significantly increased AST and ALT levels compared to the normal diet group, and blood triglyceride levels also significantly increased.

[0145] On the other hand, in the beLP1 administration group, ALT and AST levels decreased by approximately 36% and 16%, respectively, compared to the HFD group, and triglyceride levels also decreased significantly by approximately 36%.

[0146] The above results suggest that the administration of beLP1 is effective in suppressing liver dysfunction and lipid metabolism abnormalities caused by a high-fat diet.

[0147] As such, it has been confirmed that the beLP1 strain can contribute to the improvement of overall obesity-related metabolic diseases by alleviating not only weight gain but also liver dysfunction and abnormal elevation of blood lipids induced by a high-fat diet.

[0148] Experimental Example 3: Measurement of Adipose Tissue and Liver Weight

[0149] Using a high-fat diet-induced obese mouse model, the inhibitory effects of the beLP1-derived active ingredient on fat accumulation and hepatomegaly were evaluated.

[0150] The experimental groups consisted of a normal diet group, a high-fat diet group, and a high-fat diet + beLP1 administration group. After the experiment was completed, mice were sacrificed, and abdominal adipose tissue and liver tissue were extracted to measure their absolute weight and relative weight to body weight (% / BW).

[0151] Referring to Figure 3, in the HFD group, both adipose tissue and liver weight increased significantly compared to the normal diet group, which reflects the typical characteristics of fat accumulation and liver enlargement associated with a high-fat diet.

[0152] Liver weight in the HFD group was 1,911 mg, which was significantly increased compared to the normal diet group, and this is interpreted as a typical response to liver fat deposition due to a high-fat diet.

[0153] On the other hand, in the HFD + beLP1 administration group, the liver weight was 1,556 mg, which was lower than that of the HFD group, showing that beLP1 can contribute to inhibiting liver enlargement and fat accumulation.

[0154] In the case of subcutaneous fat, the HFD group showed a significant increase to the level of 3,267 mg, while the beLP1 administration group was measured at a significantly lower level of 2,556 mg, clearly demonstrating an inhibitory effect on body fat accumulation.

[0155] Meanwhile, epididymal fat remained at a significantly higher level compared to the normal diet group in both the HFD group and the HFD + beLP1 group, but showed a decreasing trend in the beLP1 administration group (2,938 mg) compared to the HFD group (3,155 mg). Similarly, pericardial fat increased in the HFD group (152 mg), but the beLP1 administration group showed a lower value (126 mg).

[0156] The above results suggest that beLP1 possesses physiological activity capable of effectively inhibiting high-fat diet-induced hepatomegaly and fat accumulation, and may contribute to alleviating the pathology of obesity-related metabolic diseases.

[0157] In particular, the significant inhibitory effect in subcutaneous fat among adipose tissues supports the possibility of linking to the inhibition of weight gain and improvement of body composition, and suggests that the composition of the present invention can be utilized as a functional material capable of inhibiting pathological conditions associated with obesity-related diseases, particularly fatty liver and metabolic syndrome.

[0158] Experimental Example 4: Human Application Test Evaluation

[0159] When heat-treated Lactobacillus plantarum beLP1 was administered, the effects on body composition, metabolic indicators, and inflammatory cytokine indicators in overweight or mildly obese adults were examined.

[0160] The study was designed with a randomized, double-blind, placebo-controlled, parallel group structure involving a total of 140 subjects, and subjects were assigned to either the beLP1 administration group (n=70) or the placebo group (n=70) after a pre-test. The administration groups were orally administered capsules containing 50 billion cells of inactivated beLP1 once a day for 12 weeks.

[0161] The initial characteristics of the test group and the control group were similar, and the average age was approximately 34.7 years and the average BMI was 29.7 kg / m².

[0162] The test was divided into Day 0 (Baseline) and Day 84 (End point) and was conducted based on a total of two visits.

[0163] The analysis items included body composition (BIA analysis), inflammatory cytokines (TNF-α, IL-6, IFN-γ), and insulin resistance indicators (HOMA-IR).

[0164] division control group experimental group Body Fat (%) -0.16% -0.45% Android Fat(%) +0.14% -0.76% Lean Body Mass(g) +239.26g +207.89g Gynoid Fat(%) -0.15% -0.46% HOMA-IR 1.39 0.61 IL-6 +10.11 +3.66 IFN-γ +18.65 +11.33 TNF-α -0.01 -0.16

[0165] Referring to Table 1, body fat percentage (Body Fat%) decreased by -0.16% in the control group, while the beLP1 administration group showed a larger decrease of -0.45%. In particular, while abdominal fat (Android Fat%) actually increased by +0.14% in the control group, it decreased by -0.76% in the beLP1 group, suggesting that beLP1 can contribute to the improvement of central obesity.

[0166] Gynoid fat percentage also decreased by -0.46% in the beLP1 administration group, showing a significant improvement compared to the control group (-0.15%).

[0167] Lean Body Mass increased in both groups, but as of day 84, the beLP1 group increased by +207.89g and the control group by +239.26g, showing that fat reduction and muscle preservation accompanied weight maintenance or slight increase.

[0168] Meanwhile, HOMA-IR, an indicator of insulin resistance, increased to 1.39 in the control group, whereas the beLP1 administration group showed a relatively lower increase to 0.61, indicating a trend of alleviation in insulin resistance, which suggests the possibility of improvement in obesity-related metabolic disorders.

[0169] Improvements following beLP1 administration were also confirmed in inflammatory cytokine markers; while IL-6 increased by +10.11 in the control group, the beLP1 group showed a lower increase of +3.66, and TNF-α decreased by -0.16 in the beLP1 group, demonstrating a clear inhibitory effect compared to the control group (-0.01).

[0170] In addition, IFN-γ increased in both groups, but the beLP1 group showed a tendency to suppress immune hyperactivity, with +11.33 being lower than the control group (+18.65).

[0171] The above results support the fact that beLP1-derived components may possess complex physiological benefits, including not only body composition improvement effects such as body fat reduction and muscle preservation, but also the alleviation of insulin resistance and chronic low-grade inflammatory conditions, suggesting that the beLP1 strain can be utilized as a functional material effective for anti-obesity, anti-inflammation, and maintaining metabolic homeostasis.

[0172] Experimental Example 5: Evaluation of Adipogenesis Inhibition Effect

[0173] The effect of the Lactobacillus plantarum beLP1 strain on inhibiting adipogenesis was evaluated.

[0174] The test used 3T3-L1 mouse preadipocyte cells and compared the degree of fat accumulation according to the treatment of each test group along with the induction of adipocyte differentiation.

[0175] beLP1 strain or other types of strains were used as the experimental group (Table 2).

[0176] division detail Example 1 beLP1 culture L. plantarum Use the supernatant after sonication and centrifugation of the beLP1 strain culture. Example 2 beLP1 dead cells Use dead cells after heat treatment, centrifugation, and drying of the beLP1 strain culture. Comparative Example 1 L. plantarum purchased from the market L. plantarum strain usage Comparative Example 2 L. paracasei Other species Lactobacillus Use of strain family Comparative Example 3 B. longum Bifidobacterium Use of strain family

[0177] Adipocyte differentiation was induced in 3T3-L1 cells using a differentiation induction medium (DMEM + IBMX, dexamethasone, insulin) under confluent conditions, and the heat-treated inactivated cells of each test group were 1×10 8 Treatment was performed at a concentration of cells / mL for 8 days starting from the start of differentiation induction.

[0178] After the experiment, Oil Red O staining was performed to evaluate intracellular triglyceride accumulation, and the stained lipids were eluted with 100% isopropanol and quantified by measuring the absorbance at 520 nm (Table 3).

[0179] division Fat accumulation inhibition rate (%) Example 1 43% Example 2 46% Comparative Example 1 16% Comparative Example 2 13% Comparative Example 3 12% Control group (untreated) 0%

[0180] Referring to Table 3, Lactobacillus plantarum The culture of beLP1 (Example 1) and the dead cell (Example 2) showed fat accumulation inhibition rates of 43% and 46%, respectively, demonstrating a significant effect in inhibiting the accumulation of triglycerides in adipocytes.

[0181] On the other hand, Comparative Example 1 ( L. plantarum In the case of strain), an inhibition rate of 16% was observed, which remained at a significantly lower level compared to the beLP1 strain, and Comparative Example 2 and Comparative Example 3 Lactobacillus paracasei and Bifidobacterium longum They also showed relatively low inhibitory effects of 13% and 12%, respectively.

[0182] The above results show that under the same conditions, the beLP1 strain is homologous to common lactic acid bacteria. L. plantarum It indicates superior ability to inhibit adipocyte differentiation compared to strains, suggesting that the beLP1 strain of the present invention possesses differentiated activity as a functional material for anti-obesity purposes.

[0183] Experimental Example 6: Evaluation of Triglyceride Accumulation Inhibitory Activity

[0184] We evaluated whether the Lactobacillus plantarum beLP1 strain has activity that inhibits triglyceride accumulation in 3T3-L1 adipocytes.

[0185] After seeding 3T3-L1 preadipocytes into a 6-well plate, once the cells reached a confluent state, differentiation was induced by treating with differentiation induction medium (DMEM + 10% FBS + dexamethasone, IBMX, insulin). Samples for each experimental group were applied starting from the start of differentiation induction, and the cells were cultured for 8 days.

[0186] The experimental group was set up in the same way as in Experimental Example 6 (Table 2), and all samples were treated at a final concentration of 100 μg / mL.

[0187] After the end of culture, the cells were washed with PBS, collected, lysed with Triton X-100 lysate, reacted according to the manufacturer's instructions using a commercial triglyceride quantification assay kit, and the absorbance was measured at 570 nm.

[0188] The triglyceride content of each group was calculated by normalizing it to the protein content, and the relative accumulation inhibition rate compared to the control group was calculated (Table 4).

[0189] division Triglyceride accumulation inhibition rate (%) Example 1 39% Example 2 41% Comparative Example 1 18% Comparative Example 2 9% Comparative Example 3 14% Control group (untreated) 0%

[0190] Referring to Table 4, Example 1 showed a triglyceride accumulation inhibition rate of 39%, and Example 2 showed 41%, which is interpreted as a very significant inhibitory effect compared to the control group (untreated) at 0%.

[0191] On the other hand, Comparative Example 1 showed an inhibition rate of 18%, which is less than half the level of beLP1, suggesting that there are distinct functional differences between strains even within the same species.

[0192] In addition, Comparative Examples 2 and 3 showed a low inhibition rate of less than 20%, confirming that only the beLP1 strain has relatively strong inhibitory activity on triglyceride accumulation.

[0193] The above results demonstrate that the beLP1-derived composition can effectively regulate lipid metabolism by acting directly on the triglyceride biosynthesis or accumulation pathways. In particular, the fact that it maintains high activity even in non-live forms, such as inactivated cells, suggests that it is a highly excellent functional material in terms of industrial utility and stability.

[0194] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single unit may be implemented in a distributed manner, and components described as distributed may likewise be implemented in a combined form.

[0195] The scope of the present invention is defined by the claims set forth below, and all modifications or variations derived from the meaning and scope of the claims and equivalent concepts thereof should be interpreted as being included within the scope of the present invention.

[0196] Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resource Center (KCTC) Trustee Number: KCTC15902BP Date of Deposit: 2024-05-07

Claims

Claim 1 Lactobacillus plantarum ( Lactobacillus plantarum A pharmaceutical composition for the prevention or treatment of obesity-related diseases selected from the group consisting of fatty liver and lipid-related metabolic syndrome, comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells, through activity of inhibiting adipocyte differentiation and inhibiting triglyceride accumulation. Claim 2 delete Claim 3 Lactobacillus plantarum ( Lactobacillus plantarum A health functional food composition for improving obesity-related diseases through the inhibition of adipocyte differentiation and triglyceride accumulation activity, comprising one or more active ingredients selected from the group consisting of the beLP1 strain (accession number: KCTC15902BP), its lysate, its culture, its concentrate, its dried product, and its dead cells. Claim 4 A health functional food composition according to paragraph 3, wherein the obesity-related disease is one or more selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome. Claim 5 Lactobacillus plantarum ( Lactobacillus plantarum A health functional food composition for anti-obesity containing one or more active ingredients selected from the group consisting of ) beLP1 strain (accession number: KCTC15902BP), lysate thereof, culture thereof, concentrate thereof, dried thereof, and dead cells thereof, with activity of inhibiting adipocyte differentiation and triglyceride accumulation. Claim 6 Lactobacillus plantarum ( Lactobacillus plantarum A health functional food composition for weight loss through activity of inhibiting adipocyte differentiation and inhibiting triglyceride accumulation, comprising one or more active ingredients selected from the group consisting of ) beLP1 strain (accession number: KCTC15902BP), lysate thereof, culture thereof, concentrate thereof, dried thereof, and dead cells thereof. Claim 7 A health functional food composition that inhibits liver fat accumulation in paragraph 6.

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