Composition for preventing or improving diabetes or obesity comprising horse oil fermented product as an active ingredient

Abstract

The present invention relates to a food composition, pharmaceutical composition, quasi-drug composition, and feed composition for the prevention or improvement of diabetes or obesity, comprising a fermented horse oil product as an active ingredient.

Description

Technology Field

[0001] The present invention relates to a composition comprising a fermented horse oil product as an active ingredient. Background Technology

[0003] Alpha-glucosidase (α-glucosidase) is an enzyme present in the epithelial cells of the small intestine that plays a role in breaking down polysaccharide and disaccharide carbohydrates into monosaccharides for digestion and absorption. Therefore, alpha-glucosidase inhibitors can manage postprandial hyperglycemia by delaying the digestion and absorption of dietary carbohydrates, thereby reducing the rise in blood glucose and blood insulin levels, and can also manage metabolic diseases such as obesity and hyperlipidemia. Furthermore, alpha-glucosidase inhibitors are utilized as important therapeutic agents in the management of type 2 diabetes.

[0004] Representative alpha-glucosidase inhibitors include acarbose, voglibose, and miglitol, but long-term use can cause serious side effects such as vomiting, diarrhea, and abdominal distension. Therefore, there is a growing need for research on replacing them with natural materials that are safer and have excellent efficacy in lowering blood sugar after meals.

[0005] Various inhibitors are being developed to achieve effects such as blood glucose lowering and obesity treatment by inhibiting the activity of alpha-glucosidase. In Korean Registered Patent No. 10-1607532, Weissella Confusah WIKIM29 ( Weissella confusa It has been registered as a pharmaceutical composition for the prevention and treatment of diabetes or obesity by inhibiting alpha-glucosidase activity containing WIKIM29).

[0006] Meanwhile, horse oil is generally known as a cosmetic ingredient capable of supplying unsaturated fatty acids to the skin because its main components are very similar to human sebum, it has high affinity for human skin, and it contains 60-65% unsaturated fatty acids. Accordingly, the inventors of the present invention researched horse oil and completed the present invention. Prior art literature

[0008] Korean Published Patent Application No. 10-2012-0044105 (Published May 7, 2012) Korean Registered Patent Application No. 10-1607532 (Published March 31, 2016) The problem to be solved

[0009] The problem that the present invention aims to solve is to provide a food composition for the prevention or improvement of diabetes or obesity comprising a fermented horse oil product as an active ingredient.

[0010] Another problem that the present invention aims to solve is to provide a pharmaceutical composition for the prevention, improvement, or treatment of diabetes or obesity comprising the above-mentioned fermented horse oil as an active ingredient.

[0011] Another problem that the present invention aims to solve is to provide a quasi-drug composition for the prevention, improvement, or treatment of diabetes or obesity that includes the above-mentioned fermented horse oil as an active ingredient.

[0012] Another problem that the present invention aims to solve is to provide a cosmetic composition for the prevention or improvement of obesity comprising the above-mentioned fermented horse oil as an active ingredient.

[0013] The problem that the present invention aims to solve is to provide a feed composition for the prevention or improvement of diabetes or obesity comprising a fermented horse oil product as an active ingredient. means of solving the problem

[0015] One embodiment of the present invention provides a food composition for the prevention or improvement of diabetes or obesity comprising a fermented horse oil product as an active ingredient.

[0016] In one embodiment of the present invention, the food composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0017] In one embodiment of the present invention, the food composition may inhibit the absorption of sugar or fat. For example, the sugar or fat may be consumed as food.

[0018] In one embodiment of the present invention, the food composition may further include uses for maintaining or improving the balance of intestinal microorganisms.

[0019] In one embodiment of the present invention, the food composition may be formed into any one formulation selected from a beverage, granules, tablets, powder, pills, and capsules.

[0020] One embodiment of the present invention provides a pharmaceutical composition for the prevention, improvement, or treatment of diabetes or obesity comprising the above-mentioned fermented horse oil as an active ingredient.

[0021] In one embodiment of the present invention, the pharmaceutical composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0022] In one embodiment of the present invention, the pharmaceutical composition may further include uses for maintaining or improving the balance of intestinal microorganisms.

[0023] One embodiment of the present invention provides a quasi-drug composition for the prevention, improvement, or treatment of diabetes or obesity, comprising the above-mentioned fermented horse oil as an active ingredient.

[0024] In one embodiment of the present invention, the quasi-drug composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0025] In one embodiment of the present invention, the quasi-drug composition may further include use for maintaining or improving the balance of intestinal microorganisms.

[0026] One embodiment of the present invention provides a cosmetic composition for the prevention or improvement of obesity comprising the above-mentioned fermented horse oil as an active ingredient.

[0027] In one embodiment of the present invention, the cosmetic composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0028] One embodiment of the present invention provides a feed composition for the prevention or improvement of diabetes or obesity comprising a fermented horse oil product as an active ingredient.

[0029] In one embodiment of the present invention, the feed composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0030] In one embodiment of the present invention, the feed composition may further include uses for maintaining or improving the balance of intestinal microorganisms.

[0031] In one embodiment of the present invention, the fermented horse oil may be prepared by mixing barley koji with oil refined after melting horse fat meat and then fermenting it.

[0032] In one embodiment of the present invention, the fermented horse oil may be prepared by mixing the horse oil with barley koji in a weight ratio of 5:1 to 1:5 and fermenting it.

[0033] In one embodiment of the present invention, the fermented horse oil may be fermented for 10 days or more.

[0034] In one embodiment of the present invention, the fermented horse oil may be one in which the microbial community within the horse oil has changed due to fermentation.

[0035] In one embodiment of the present invention, the change in the microbial community may be one or more of the following: a decrease in the population of pathogenic bacteria or opportunistic bacteria as fermentation proceeds; and an increase in the population of beneficial bacteria.

[0036] In one embodiment of the present invention, the pathogenic bacteria or opportunistic bacteria are Salmonella enterica ( Salmonella enterica ), Enterobacter quacihormaechei( Enterobacter quasihormaechei ), Atlantibacter Hermani( Atlantibacter hermanni ), Enterococcus faecalis( Enterococcus faecalis ), Enterococcus faecium( Enterococcus faecium ), Enterococcus galinarum( Enterococcus fowleri ), Enterococcus hirae ( Enterococcus hirae ), Enterococcus inesi ( Enterococcus innesii ), Enterobacter covey ( Enterobacter kobei ) and Staphylococcus warneri Staphylococcus warneri It may include one or more bacteria selected from a group consisting of ).

[0037] In one embodiment of the present invention, the opportunistic bacteria is Leuconostoc mesenteroroides ( Leuconostoc mesenteroides ), Weissella paramesenteroides( Weissella paramesenteroides ) and Clostridium butyricum ( Clostridium butyricum It may include one or more bacteria selected from a group consisting of ).

[0038] In one embodiment of the present invention, the beneficial bacteria is Lactobacillus acidophilus ( Lactobacillus acidophilus ), Acetobacter persis ( Acetobacter persica ), Bacillus subtilis Bacillus subtilis ), Bifidobacterium animalis( Bifidobacterium animalum ), Clostridium diolis( Clostridium diolis ), Enterococcus mundti( Enterococcus mundtii ), Gluconobacter japonicus ( Gluconobacter japonicus ), Lactococcus lactis( Lactococcus lactis ) and Phytobacter palmae( Phytobacter palm It may include one or more bacteria selected from a group consisting of ).

[0039] One embodiment of the present invention provides a method for treating diabetes or obesity, comprising the step of administering to a human a composition containing a fermented horse oil product as an active ingredient.

[0040] One embodiment of the present invention provides a method for treating diabetes or obesity, comprising the step of administering the composition, which includes a fermented horse oil product as an active ingredient, to an animal other than a human.

[0041] One embodiment of the present invention provides a method for preventing diabetes or obesity, comprising the step of administering to a human a composition containing a fermented horse oil product as an active ingredient. Effects of the invention

[0043] Since the fermented horse oil of the present invention has excellent alpha-glucosidase inhibitory efficacy, it has the advantage of being useful as a food composition for preventing or improving diabetes or obesity; a pharmaceutical composition or quasi-drug composition for preventing, improving, or treating diabetes or obesity; a cosmetic composition for preventing or improving obesity; or a feed composition for preventing or improving diabetes or obesity.

[0044] The fermented horse oil of the present invention has the advantage of being useful as a food composition, pharmaceutical composition, quasi-drug composition, or feed composition for maintaining or improving intestinal microbial balance, as the number of pathogenic or opportunistic bacteria in the horse oil decreases and the number of beneficial bacteria increases due to fermentation. Brief explanation of the drawing

[0046] Figure 1(a) shows human maltase-glucoamylase, Figure 1(b) shows the binding interaction of MGAM-oleic acid amide, Figure 1(c) shows the binding interaction of MGAM-palmitate amide, and Figure 1(d) shows the binding interaction of MGAM-stearic acid amide. Figure 2(a) shows the alpha-glucosidase inhibitory activity of the control acarbose, and Figure 2(b) shows the alpha-glucosidase inhibitory activity of the comparative example (HO), example 1 (HOF3), example 2 (HOF5), and example 10 (HOF10). Specific details for implementing the invention

[0047] The present invention will be described in more detail below.

[0048] The following specific functional descriptions are merely illustrative to explain embodiments according to the concept of the present invention, and embodiments according to the concept of the present invention may be implemented in various forms and should not be interpreted as being limited to the embodiments described herein.

[0049] Since embodiments according to the concept of the present invention may be subject to various modifications and may take various forms, specific embodiments are to be described in detail in this specification. However, this is not intended to limit the embodiments according to the concept of the present invention to specific disclosed forms, and it should be understood that they include all modifications, equivalents, and substitutions that fall within the spirit and scope of the present invention.

[0050] The terms used in this specification are used merely to describe specific embodiments and are not intended to limit the invention. Singular expressions include plural expressions unless the context clearly indicates otherwise.

[0051] 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 specification.

[0052] Throughout this specification, '%' used to indicate the concentration of a particular substance is (w / w) % for solid / solid, (w / v) % for solid / liquid, and (v / v) % for liquid / liquid, unless otherwise noted.

[0053] One embodiment of the present invention provides a food composition for the prevention or improvement of diabetes or obesity comprising a fermented horse oil product as an active ingredient.

[0054] The above horse oil is referred to as horse fat, horse fat, or horse oil, and may be oil (refined horse oil) obtained by melting and refining the horse's fatty meat or fatty tissue. Preferably, the horse's fatty meat can be finely chopped, melted on a hot plate set to 60–100°C, and then filtered using filter paper inside a drying oven. The horse oil filtered through the above process can be further filtered using filter paper mixed with Celite and activated carbon. Through this additional filtration process, the characteristic odor of the horse is more completely removed, and horse oil that is close to white can be produced. Furthermore, this additional filtration process may be performed repeatedly. Since the color and odor of the fatty meat of a slaughtered horse vary significantly depending on the horse's condition, age, and the specific part of the meat, this point must be fully understood when selecting the fatty meat. If horse fat meat that is close to yellow in color and has a strong odor is selected, even after the filtration process, the yellow color remains and the odor is strong, so it may not be suitable for selection as a food composition, pharmaceutical composition, quasi-drug composition, or cosmetic composition.

[0055] In the present invention, the term “purification” may be used interchangeably with “obtaining,” “isolating,” “extracting,” or “recovering.”

[0056] In the present invention, the term “diabetes mellitus” refers to a type of metabolic disease characterized by high blood glucose levels, such as insufficient insulin secretion or failure to perform normal function. The above-mentioned diabetes is classified into Type 1 diabetes and Type 2 diabetes. Type 1 diabetes is a disease caused by the inability to produce any insulin, while Type 2 diabetes is characterized by insulin resistance (a condition in which the function of insulin to lower blood sugar is reduced, preventing cells from effectively burning glucose).

[0057] In the present invention, the diabetes may be Type 1 diabetes or Type 2 diabetes, and the terms "diabetes" or "diabetes" may be used interchangeably.

[0058] In this invention, the term “obesity” refers to a state of energy imbalance within the human body resulting from consuming more energy than is required or consuming less energy than is consumed. Such obesity is a cause of various metabolic diseases, such as cardiovascular disease, hypertension, and type 2 diabetes. In cases of obesity, fat cells store lipids as a long-term energy source and secrete various adipokines involved in the metabolism and inflammation of both fat and non-fat cells, thereby inducing chronic inflammation and causing related metabolic diseases such as insulin resistance.

[0059] In one embodiment of the present invention, the food composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0060] In one embodiment of the present invention, the food composition may inhibit the absorption of sugar or fat. For example, the sugar or fat may be consumed as food.

[0061] Inhibiting the above-mentioned alpha-glucosidase delays the digestion and absorption of carbohydrates, thereby preventing a rapid rise in blood sugar levels and reducing the rise in blood sugar after meals. Additionally, it helps to suppress weight gain by reducing insulin secretion, which can help prevent, improve, or treat diabetes or obesity.

[0062] The fermented horse oil of the present invention has excellent alpha-glucosidase inhibitory activity, so it has the advantage of being useful as a food composition for preventing or improving diabetes or obesity, a pharmaceutical composition or quasi-drug composition for preventing, improving, or treating diabetes or obesity, a cosmetic composition for preventing or improving obesity, or a feed composition for preventing or improving diabetes or obesity.

[0063] In the present invention, “included as an active ingredient” means including an amount sufficient to produce desirable biological effects, such as exhibiting alpha-glucosidase (α-glucosidase) inhibitory efficacy, exhibiting a preventive or improving effect on diabetes, exhibiting an anti-obesity effect, exhibiting a body fat reduction effect, exhibiting a preventive or improving effect on obesity, or beneficial results not limited thereto, and this amount is referred to as the “effective amount.” Accordingly, the effective amount will vary depending on the form in which the active ingredient is manufactured, the method of application to food, pharmaceuticals, quasi-pharmaceuticals, or feed, and the time spent on the skin.

[0064] In one embodiment of the present invention, the fermented horse oil may be prepared by mixing barley koji with oil refined after melting horse fat meat and then fermenting it.

[0065] The above barley koji is made of yellow koji mold (Aspergillus oryzae, Aspergillus oryzae It is a product in which barley is fermented using ) as a fermentation bacterium, and it may be used as a product made directly or purchased commercially. The above barley koji is not limited to any ordinary barley koji used in the industry for fermenting horse milk.

[0066] The above Aspergillus oryzae ( Aspergillus oryzae It is a known strain, and its optimal growth temperature is typically around 30°C. It prefers high temperature and high humidity, and the organic acids produced by Aspergillus oryzae are kojic acid, gluconic acid, etc.

[0067] In one embodiment of the present invention, the fermented horse oil may be prepared by mixing barley koji with the horse oil in a weight ratio of 5:1 to 1:5 and fermenting it. Specifically, the fermented horse oil may be prepared by mixing barley koji with the horse oil in a weight ratio of 3:1 to 1:3 and fermenting it.

[0068] In one embodiment of the present invention, the fermented horse oil may be prepared by mixing barley koji with the horse oil in a weight ratio of 5:1 to 1:5 and fermenting it for 1 day or more, 3 days or more, 5 days or more, or 10 days or more.

[0069] In one embodiment of the present invention, the fermented horse oil may have a change in the microbial community within the horse oil due to fermentation.

[0070] In one embodiment of the present invention, the change in the microbial community may be a decrease in the population of pathogenic bacteria or opportunistic bacteria and / or an increase in the population of beneficial bacteria as fermentation proceeds.

[0071] In the present invention, the term “pathogenic bacteria” refers to bacteria that cause disease, which infect a host, produce strong toxins, or directly attack the immune system. For example, pathogenic bacteria include Salmonella enterica, which causes food poisoning and typhoid fever. Salmonella enterica ), tuberculosis bacteria that cause tuberculosis ( Mycobacterium tuberculosis There are ) etc.

[0072] In this invention, the term “opportunistic bacteria” refers to bacteria that cause infection in specific environments, specifically bacteria that cause infection when the host’s immunity is lowered or the body’s balance is disrupted. For example, opportunistic bacteria include E. coli ( Escherichia coli E. coli usually exists as a commensal bacterium in the intestines and does not cause disease, but when the host's immunity is lowered, some strains of E. coli cause urinary tract infections or enteritis.

[0073] In this invention, the term “beneficial bacteria” refers to bacteria that exert beneficial effects on humans, inhibit the growth of harmful intestinal bacteria, and contribute to the enhancement of immunity. Beneficial bacteria are essential for maintaining the balance of the intestinal microbiome and play roles such as immune regulation, digestive aid, vitamin synthesis, and inhibition of pathogens. For example, beneficial bacteria include Lactobacillus acidophilus, a lactic acid bacterium that maintains intestinal health ( Lactobacillus acidophilus Bacillus subtilis, which breaks down proteins and maintains the balance of intestinal microorganisms Bacillus subtilis There are ) etc.

[0074] In one embodiment of the present invention, the pathogenic bacteria or opportunistic bacteria are Salmonella enterica ( Salmonella enterica ), Enterobacter quacihormaechei( Enterobacter quasihormaechei ), Atlantibacter Hermani( Atlantibacter hermanni ), Enterococcus faecalis( Enterococcus faecalis ), Enterococcus faecium( Enterococcus faecium ), Enterococcus galinarum( Enterococcus fowleri ), Enterococcus hirae ( Enterococcus hirae ), Enterococcus inesi ( Enterococcus innesii ), Enterobacter covey ( Enterobacter kobei ) and Staphylococcus warneri Staphylococcus warneri It may include one or more bacteria selected from a group consisting of ).

[0075] The above Salmonella enterica ( Salmonella enterica Salmonella enterica is a pathogenic bacterium and is a major causative agent of systemic infections such as food poisoning, typhoid fever, and sepsis. Salmonella enterica inhabits the intestines of humans and animals, as well as contaminated food and water.

[0076] The above Enterobacter quasihormaechii ( Enterobacter quasihormaechei ) is an opportunistic bacterium that can cause infection in immunocompromised individuals or hospitalized patients. Other Enterobacter Similar to the disease, it can cause urinary tract infections, respiratory infections, bloodstream infections, etc.

[0077] The above Atlantibacter Hermani ( Atlantibacter hermanni It is an opportunistic bacterium that is generally less pathogenic but can cause wound infections, eye infections, and bloodstream infections in immunocompromised individuals or patients using medical devices. Atlantibacter hermannii can be isolated from various environments, including water, soil, food, and clinical samples.

[0078] The above Enterococcus faecalis ( Enterococcus faecalis Enterococcus faecalis is an opportunistic bacterium that exists as part of the normal microbiome in the intestines of humans and animals, and some strains can be used as probiotics. However, Enterococcus faecalis can cause infections when immunity is weakened or when the intestinal balance is disrupted following antibiotic use; in particular, Enterococcus faecalis is one of the major causative agents of sepsis, urinary tract infections, endocarditis, and peritonitis in hospital settings. Enterococcus faecalis inhabits the intestines (small and large intestines), oral cavity, and vagina of humans and animals.

[0079] The above Enterococcus faecium ( Enterococcus faeciumEnterococcus faecium is an opportunistic bacterium; some strains are used as probiotics and can contribute to maintaining gut health and regulating immunity, but it can cause infections when immunity is weakened or after antibiotic use. In particular, Enterococcus faecium can cause VRE (Vancomycin-Resistant Enterococcus faecium) infections in hospital settings. Enterococcus faecium inhabits the intestines, skin, and mucous membranes of humans and animals.

[0080] The above Enterococcus galinarum ( Enterococcus fowleri ) is an opportunistic bacterium that is naturally present in the intestines of healthy humans and animals, but can cause bloodstream infections (bacteremia, sepsis), endocarditis, urinary tract infections, etc. when immunity is compromised.

[0081] The above Enterococcus hirae ( Enterococcus hirae Enterococcus hirae is an opportunistic bacterium that is generally found in the intestines of animals (especially poultry, cattle, and pigs), and some are also found in the human intestine. In immunocompromised individuals and hospital settings, Enterococcus hirae can cause sepsis, endocarditis, urinary tract infections, peritonitis, and bloodstream infections.

[0082] The above Enterococcus inesi ( Enterococcus innesii ) is an opportunistic bacterium found mainly in the intestines of insects, and since all strains possess the vanC-4 gene and have natural resistance to vancomycin, it can be classified as a potential opportunistic pathogen.

[0083] The above Enterobacter covey ( Enterobacter kobei Enterobacter covey is an opportunistic bacterium that can cause infections such as sepsis, urinary tract infections, respiratory infections, wound infections, and meningitis in immunocompromised individuals or hospitalized patients. Enterobacter covey can be isolated from clinical samples such as blood and urine.

[0084] The above Staphylococcus warneri Staphylococcus warneriStaphylococcus warneri is an opportunistic bacterium that inhabits the skin and mucous membranes of humans and animals. Although Staphylococcus warneri is a harmless normal bacterium on the skin and mucous membranes of healthy people, it can cause infection if immunity is weakened or if it attaches to medical devices such as catheters, artificial heart valves, and artificial joints.

[0085] In one embodiment of the present invention, the opportunistic bacteria is Leuconostoc mesenteroroides ( Leuconostoc mesenteroides ), Weissella paramesenteroides( Weissella paramesenteroides ) and Clostridium butyricum ( Clostridium butyricum It may include one or more bacteria selected from a group consisting of ).

[0086] The above Leuconostoc mesenteroroides ( Leuconostoc mesenteroides ) is a bacterium that produces lactic acid during the fermentation process. However, some strains of Leuconostoc mesenteroroides can cause infections such as bacteremia, sepsis, endocarditis, and urinary tract infections in immunocompromised and critically ill patients, so they can be classified as opportunistic bacteria.

[0087] The above Weissella paramesenteroides ( Weissella paramesenteroides ) is a bacterium that produces lactic acid bacteria in fermented foods such as kimchi, doenjang, and cheese, inhibiting harmful bacteria and promoting fermentation, and helping to maintain the balance of intestinal microorganisms and the proliferation of beneficial bacteria. Generally, Weissella paramesenteroides is a harmless bacterium, but since it can cause bacteremia or sepsis in immunocompromised patients, it can be classified as an opportunistic bacterium.

[0088] The above Clostridium butyricum ( Clostridium butyricum Clostridium butyricum is a bacterium that produces butyric acid in the intestines, providing an energy source for intestinal epithelial cells and protecting the intestinal barrier. Generally, Clostridium butyricum is a harmless bacterium, but certain strains can cause bloodstream infections (bacteremia), intestinal infections, and sepsis, so they can be classified as opportunistic bacteria.

[0089] In one embodiment of the present invention, the beneficial bacteria is Lactobacillus acidophilus ( Lactobacillus acidophilus ), Acetobacter persis ( Acetobacter persica ), Bacillus subtilis Bacillus subtilis ), Bifidobacterium animalis( Bifidobacterium animalum ), Clostridium diolis( Clostridium diolis ), Enterococcus mundti( Enterococcus mundtii ), Gluconobacter japonicus ( Gluconobacter japonicus ), Lactococcus lactis( Lactococcus lactis ) and Phytobacter palmae( Phytobacter palm It may include one or more bacteria selected from a group consisting of ).

[0090] The above Lactobacillus acidophilus ( Lactobacillus acidophilus Lactobacillus acidophilus is a beneficial bacterium commonly found in the intestines, oral cavity, and vagina of humans and animals. It is a strain widely used as a probiotic because it ferments lactose, which is essential for dairy production, into lactic acid. Lactobacillus acidophilus is effective in maintaining intestinal health and strengthening immunity.

[0091] The above Acetobacter persis ( Acetobacter persica Acetobacter persisi is a beneficial bacterium that oxidizes ethanol to produce acetic acid and is a strain utilized in fermentation industries such as vinegar production. Acetobacter persisi can be isolated from sugarcane stalks, fruits, flowers, etc.

[0092] The above Bacillus subtilis ( Bacillus subtilis ) is a beneficial bacterium that inhibits harmful bacteria in the intestines and aids digestion by producing digestive enzymes such as amylase and protease, and some strains are used industrially for bioplastic production and environmental purification.

[0093] The above Bifidobacterium animalis ( Bifidobacterium animalum ) is a beneficial bacterium that produces lactic acid and acetic acid in the intestines to lower intestinal acidity and Clostridium difficile , Salmonella It is a strain that inhibits the proliferation of harmful bacteria. Bifidobacterium animalis is effective in regulating the immune system, activating immune responses, and reducing infections and inflammatory reactions by maintaining the balance of intestinal microorganisms.

[0094] The above Clostridium diolis ( Clostridium diolis Clostridium diolis is a beneficial bacterium that ferments glycerol to produce 1,3-propanediol. Clostridium diolis can be isolated from soil, water, fermentation environments, etc.

[0095] The above Enterococcus mundti ( Enterococcus mundtii )Is It is possible to produce bacteriocins such as mundticin and enterocin, and Listeria monocytogenes , Enterococcus faecalis It can be classified as a beneficial bacterium because it inhibits harmful bacteria. Enterococcus mundti inhabits the intestines of humans and animals, soil, water, and the like.

[0096] The above Gluconobacter japonica ( Gluconobacter japonicus Gluconobacter japonicus is a beneficial bacterium that partially oxidizes various substrates such as glucose and ethanol to produce organic acids such as gluconic acid and ketogluconic acid, and is a strain that produces various organic acids, including 2-ketogluconic acid, a precursor of vitamin C. Gluconobacter japonicus can be isolated from sugar-rich environments such as fruits and flowers.

[0097] The above Lactococcus lactis ( Lactococcus lactis Lactococcus lactis is a beneficial bacterium that produces lactic acid in the intestines to suppress harmful bacteria, promotes the proliferation of beneficial bacteria, and regulates the immune system. Lactococcus lactis produces the bacteriocin nisin, Listeria monocytogenes It has the effect of inhibiting food poisoning bacteria.

[0098] The above Phytobacter palmae ( Phytobacter palm) is a strain that fixes atmospheric nitrogen and converts it into a form available to plants. Phytobacter palma can be classified as beneficial bacteria because it performs beneficial roles through symbiotic relationships with plants, such as producing plant hormones and suppressing pathogens. Phytobacter palma includes oil palm ( Guinean olive It can be isolated from the leaf tissue of ).

[0099] In one embodiment of the present invention, the food composition may further include uses for maintaining or improving the balance of intestinal microorganisms.

[0100] The “microbial balance” of the present invention refers to a state in which pathogenic, opportunistic, and beneficial microorganisms are distributed in balance within a microbial community. If this microbial balance is disrupted by external factors, it may have harmful effects on the body or individual. Furthermore, if the microbial imbalance becomes severe, pathogens, toxins, etc., may stimulate the immune system, thereby causing inflammatory diseases or autoimmune diseases. For example, long-term use of antibiotics can disrupt the balance of intestinal microorganisms and induce the overgrowth of specific pathogens, which can increase inflammatory responses.

[0101] In the present invention, “maintenance of microbial balance” refers to restoring or maintaining the distribution of pathogenic microorganisms, opportunistic microorganisms, and beneficial microorganisms in a body or individual when in a normal state, and “improvement of microbial balance” refers to a decrease in the proportion of pathogenic or opportunistic microorganisms and an increase in the proportion of beneficial microorganisms within the entire microbial community.

[0102] The fermented horse oil of the present invention has the advantage of being useful as a food composition, pharmaceutical composition, quasi-drug composition, or feed composition for maintaining or improving the balance of intestinal microorganisms, as the number of pathogenic or opportunistic bacteria in the horse oil decreases and the number of beneficial bacteria increases due to fermentation.

[0103] In one embodiment of the present invention, the food composition may be formed into any one formulation selected from a beverage, granules, tablets, powder, pills, and capsules.

[0104] There are no restrictions on the types of the above foods, and they are not limited to any products that may contain or be added to the fermented horse oil of the present invention, such as sausages, meat, bread, chocolates, snacks, candies, confectionery, ramen, pizza, other noodles, chewing gum, ice cream, dairy products, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

[0105] Specifically, the above food composition may be a health functional food. The above health functional food is a food that emphasizes the bio-regulatory function of food and is a food to which added value is imparted by utilizing physical, biochemical, or biotechnological methods to act and manifest for a specific purpose. The components of such health functional food are designed and processed to fully exert bio-regulatory functions on the body related to biological defense, regulation of body rhythms, and prevention and recovery from disease, and may contain food additives, sweeteners, or functional ingredients that are acceptable as food.

[0106] The above-mentioned health functional food may be manufactured and processed into any one formulation selected from the group consisting of beverages, granules, tablets, powders, capsules, liquid solutions, and pills for the purpose of improving diabetes or improving obesity.

[0107] When the fermented horse oil of the present invention is used as a health functional food (or an additive for a health functional beverage), it may be added as is, used in combination with other foods or food ingredients, or used appropriately according to conventional methods. The mixing amount of the fermented horse oil may be appropriately determined according to its intended use (prevention, health or improvement, therapeutic treatment).

[0108] Additionally, specifically, the above food composition may be for inner beauty. The term “inner beauty” collectively refers to “edible cosmetics” that resolve skin problems through dietary habit improvement in the form of food while serving the purpose of cosmetics, and is not limited to those that include, add to, or mix the fermented horse oil of the present invention.

[0109] One embodiment of the present invention provides a pharmaceutical composition for the prevention, improvement, or treatment of diabetes or obesity comprising the above-mentioned fermented horse oil as an active ingredient.

[0110] In one embodiment of the present invention, the pharmaceutical composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0111] In one embodiment of the present invention, the pharmaceutical composition may further include uses for maintaining or improving the balance of intestinal microorganisms.

[0112] The efficacy of the above pharmaceutical composition is the same as that of the food composition described above.

[0113] In the present invention, the term “pharmaceuticalally acceptable” refers to that which the substance or composition is chemically and / or toxicologically compatible with the mammal and / or other components, including the formulation, that are treated by it.

[0114] In the present invention, the terms “treatment” and “improvement” refer to any act in which the symptoms of a disease are improved or beneficially altered by the administration of the pharmaceutical composition of the present invention. “Treatment” relates to any treatment that improves a health condition or extends (increases) an individual’s lifespan. The said treatment refers to any act that eradicates an individual’s disease, stops or slows down the development of an individual’s disease, suppresses or delays the development of an individual’s disease, reduces the frequency or severity of an individual’s symptoms, and / or reduces the recurrence of a person currently suffering from a disease or an individual who has previously suffered from a disease.

[0115] In the present invention, the term “prevention” refers to any act that lowers the likelihood of an individual contracting a disease or prevents or avoids symptoms of a disease in advance.

[0116] In the present invention, the terms “individual” and “subject” are used interchangeably. They refer to a human, non-human primate, or other mammal (e.g., mouse, rat, rabbit, dog, cat, cattle, pig, sheep, horse, or primate) that may or may not have a disease or disability or be infected. In many embodiments, the individual is a human. Unless otherwise specified, the terms “individual” and “subject” do not denote a specific age and include adults, the elderly, children, and newborns. In preferred embodiments of the present invention, the “individual” and “subject” are patients. Patient means a subject of treatment according to the present invention, particularly a subject with a disease.

[0117] A pharmaceutical composition according to one embodiment of the present invention may be in the form of a capsule, tablet, granule, injection, ointment, powder, or beverage.

[0118] A pharmaceutical composition according to one embodiment of the present invention may be used in the form of oral formulations such as powders, granules, capsules, tablets, and aqueous suspensions, as well as external preparations, liquids, suppositories, and injections. When formulating the pharmaceutical composition, a pharmaceutical preparation may be manufactured by mixing it with pharmaceutically acceptable excipients, adjuvants, analgesics, isotonic agents, preservatives, and other adjuvants, and formulating it into a pharmaceutically acceptable form, but is not limited thereto.

[0119] When the above pharmaceutical composition is formulated into a liquid or injectable, it may contain 10 to 40 percent of propylene glycol, etc., if necessary. The above liquid or injectable may contain any diluent or buffer known in the art. In addition, the above pharmaceutical composition may be prepared immediately before use by storing a preparation containing an active ingredient in a container such as a vial and adding a carrier or adjuvant, saline solution, etc., required for the injectable before use.

[0120] A pharmaceutical composition according to one embodiment of the present invention may further include at least one adjuvant, but is not limited thereto. The adjuvant may include aluminum hydroxide, alhydrogel, oil derived from animals or plants, mineral oil, aluminum compound, muramyl dipeptide, lipopolysaccharide (LPS), monophosphoryl lipid A (MPLA), or quil A, but is not limited thereto.

[0121] The pharmaceutical composition of the present invention can be administered to an individual in an effective amount. The “effective amount” refers to a sufficient amount capable of producing effects such as prevention, improvement, or treatment of diabetes, reduction of body fat, prevention, improvement, or treatment of obesity, and an amount that does not cause side effects or serious or excessive immune responses. The precise dosage concentration varies depending on the subject to be administered and can be easily determined by a person skilled in the art based on factors well known in the medical field, such as the age, weight, health, gender, sensitivity of the individual to drugs, route of administration, and method of administration of the subject to vaccination, and can be administered one to several times.

[0122] One embodiment of the present invention provides a quasi-drug composition for the prevention, improvement, or treatment of diabetes or obesity, comprising the above-mentioned fermented horse oil as an active ingredient.

[0123] In one embodiment of the present invention, the quasi-drug composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0124] In one embodiment of the present invention, the quasi-drug composition may further include use for maintaining or improving the balance of intestinal microorganisms.

[0125] The efficacy of the above-mentioned quasi-drug composition is the same as that of the food composition described above.

[0126] In the present invention, “quasi-drug” refers to an article falling under one of the following categories: fibers, rubber products, or similar items used for the purpose of diagnosing, treating, improving, alleviating, managing, or preventing diseases in humans or animals; items similar thereto that have a weak or non-direct effect on the human body and are not instruments or machines; and preparations used for sterilization, insecticidal, and similar purposes for the prevention of infection. It excludes articles used for the purpose of diagnosing, treating, alleviating, managing, or preventing diseases in humans or animals that are not instruments, machines, or devices, and articles used for the purpose of exerting pharmacological effects on the structure and function of humans or animals that are not instruments, machines, or devices. It also includes external preparations and personal hygiene products. More specifically, it may be, but is not limited to, disinfectants, cleansers, shower foams, mouthwash, wet wipes, detergent soaps, hand washes, or ointments.

[0127] When the fermented horse oil of the present invention is used as a quasi-drug composition, the fermented horse oil may be added as an active ingredient or appropriately used in combination with other quasi-drugs or quasi-drug ingredients according to conventional methods. The amount of the active ingredient can be appropriately determined according to the purpose of use.

[0128] In addition, the above composition may further include additives such as fragrances, colorants, disinfectants, antioxidants, preservatives, humectants, thickeners, inorganic salts, emulsifiers, and synthetic polymer materials to improve physical properties, but is not limited thereto.

[0129] Furthermore, the above composition may be administered by a spray method, a squeeze method, or by an electric machine, and may be used in any formulation form; however, it is preferable that it be in a liquid form for convenience in washing, and it may also be manufactured and used as a paste, ointment, or spray formulation, but is not limited thereto.

[0130] One embodiment of the present invention provides a cosmetic composition for the prevention or improvement of obesity comprising the above-mentioned fermented horse oil as an active ingredient.

[0131] In one embodiment of the present invention, the cosmetic composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0132] The efficacy of the above cosmetic composition is the same as that of the food composition described above.

[0133] In one embodiment of the present invention, the cosmetic composition may be a slimming cosmetic composition comprising a fermented horse oil product as an active ingredient.

[0134] The cosmetic composition of the present invention can be prepared in any formulation commonly manufactured in the art, and the cosmetic composition can be formulated into a lotion, nourishing lotion, nourishing cream, massage cream, essence, pack, paste, patch, gel, cream, lotion, powder, soap, cleanser, oil, foundation, makeup base, wax, and spray, but is not limited thereto.

[0135] In each formulation of the above cosmetic composition, other ingredients may be appropriately incorporated in addition to the essential ingredients, depending on the type of formulation or purpose of use, within a range that does not impede the purpose according to the present invention.

[0136] The above cosmetic composition may include a conventionally acceptable carrier, and may appropriately incorporate, for example, oils, water, surfactants, humectants, lower alcohols, thickeners, chelating agents, colorants, preservatives, fragrances, etc., but is not limited thereto.

[0137] The above acceptable carriers may vary depending on the formulation. For example, when formulated as an ointment, paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, or extracts thereof may be used as carrier components.

[0138] When the above cosmetic composition is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or extracts thereof may be used as carrier components, and in the case of a spray, it may further include a propellant such as chlorofluorohydrocarbon, propane, butane, or dimethyl ether.

[0139] When the above cosmetic composition is formulated as a solution or emulsion, a solvent, a solubilizing agent, or an emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan may be used.

[0140] When the above cosmetic composition is formulated as a suspension, liquid diluents such as water, ethanol, or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracanth may be used as carrier components.

[0141] When the above cosmetic composition is formulated into soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars, etc. may be used as carrier components.

[0142] The above cosmetic composition may additionally contain auxiliary agents commonly used in the fields of cosmetic science or dermatology, such as fatty substances, organic solvents, solvents, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion chelating agents, chelating agents, preservatives, blockers, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, and any other ingredients commonly used in cosmetics, depending on the quality or function of the final product.

[0143] However, the above auxiliary agent and its mixing ratio can be appropriately selected so as not to affect the desirable properties of the cosmetic composition according to the present invention.

[0144] One embodiment of the present invention provides a feed composition for the prevention or improvement of diabetes or obesity comprising a fermented horse oil product as an active ingredient.

[0145] In one embodiment of the present invention, the feed composition may have an alpha-glucosidase (α-glucosidase) inhibitory effect.

[0146] In one embodiment of the present invention, the feed composition may further include uses for maintaining or improving the balance of intestinal microorganisms.

[0147] The above feed composition may include a feed additive. The feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act.

[0148] In the present invention, the term “feed” refers to any natural or artificial prescribed food, single meal, etc., or the components of said single meal, intended for or suitable for animals to eat, consume, and digest.

[0149] The types of the above feed are not particularly limited, and feeds commonly used in the relevant technical field may be used. Non-limiting examples of the above feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, minerals, oils and fats, single-cell proteins, zooplankton, or food waste. These may be used individually or in a mixture of two or more types.

[0150] In the present invention, the term “feed additive” includes substances added to feed for the purpose of various effects, such as lowering blood sugar, preventing or improving diabetes, reducing body fat, preventing or improving obesity, supplementing nutrients, enhancing the digestibility and utilization of fiber in feed, improving milk quality, preventing reproductive disorders and improving conception rates, and preventing high-temperature stress during the summer. The feed additive of the present application corresponds to an auxiliary feed under the Feed Management Act and may additionally include mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals; mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium; vitamin preparations such as carotene, vitamin A, vitamin D, vitamin E, nicotinic acid, and vitamin B complex; protected amino acid preparations such as methionine and lysine; protected fatty acid preparations such as calcium salts of fatty acids; probiotics (lactic acid bacteria preparations); probiotics such as yeast cultures and mold fermentation products; and yeast preparations.

[0151] The feed composition according to the present application may be applied without limitation to any individual other than humans for the purpose of improving, preventing, or alleviating retinal diseases. For example, it may be applied to any individual, such as non-human animals like dogs, cats, monkeys, rabbits, guinea pigs, rats, mice, cattle, sheep, pigs, goats, etc., as well as birds and fish.

[0153] The present invention will be explained in detail below through manufacturing examples, examples, and comparative examples. However, the following manufacturing examples, examples, and comparative examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following examples.

[0155] <제조예>

[0156] 1. The manufacture of

[0157] Horse oil was purchased from Daebong LS (Jeju, Korea), and barley koji was purchased from Dongmun Traditional Market (Jeju, Korea).

[0158] To prepare the fermented horse oil, 500 mL of triple distilled water (3DW) and 50 g of sugar were placed in a 1 L Erlenmeyer flask and sterilized in an autoclave at 121°C for 15 minutes. 50 g of barley koji and 50 g of horse oil were added to the malt to prepare a mixture. The mixture was placed in an incubator (NB-203XL, N-BIOTEK, Inc., Bucheon, Korea) and fermented for a total of 10 days at 28°C and 150 rpm with stirring. During the fermentation process, 135 mL of the sample was extracted on the 3rd, 5th, and 10th days to obtain the sample. After transferring 135 mL of the fermented sample to an Erlenmeyer flask in a clean bench, an equal volume of n-hexane (135 mL) was added. A magnetic stirrer coated with 3DW was placed in the mixture, the flask was sealed with aluminum foil, and the mixture was stirred at 200 rpm for 30 minutes using the magnetic stirrer. Afterward, the resulting solution was filtered twice under gravity using No. 2 filter paper (300 mm) to collect the extracted solution. The filtered extract (500 mL) was concentrated under reduced pressure using a rotary evaporator (EYELA N-1210B, Sunil Eyela Co., Ltd., Sungnam, Korea) to collect the final sample. During concentration, the water bath was maintained at 45°C and the condenser at -10°C, and the initial pressure was set to 100 mbar and gradually decreased.

[0160] <실시예 1>

[0161] A horse oil n-hexane extract (HOF3) fermented on the 3rd day of preparation was prepared as Example 1.

[0162] <실시예 2>

[0163] The horse oil n-hexane extract (HOF5) fermented on the 5th day of preparation in the example was prepared as Example 2.

[0164] <실시예 3>

[0165] A horse oil n-hexane extract (HOF10) fermented on the 10th day of preparation was prepared as Example 3.

[0166] <비교예 1>

[0167] A horse oil n-hexane extract (HO) without barley koji in the preparation example was prepared as Comparative Example 1.

[0169] <실호예 1> 고산 아이스

[0170] Examples 1 to 3 and Comparative Example 1 were dissolved in chloroform, and ultrasonic extraction was performed for 30 minutes to ensure complete dissolution. The extracted solution was then filtered using a 0.2 μm PVDF filter to remove particles, and the filtrate was used for GC-MS / MS analysis. GC-MS / MS analysis was performed using a TQ-8050NX GC-MS / MS system (Shimadzu, Japan). Compound separation was performed using an SH-Rxi™-5Sil MS column (Shimadzu, Japan) with a length of 30 m, an inner diameter of 0.25 mm, and a film thickness of 0.25 μm. Helium was used as the carrier gas, and the flow rate was maintained at 1.56 mL / min. The injector temperature was set to 280°C, and the ion source and interface temperatures were maintained at 250°C and 300°C, respectively. Samples of Examples 1 to 3 and Comparative Example 1 were introduced into the system using a split injection mode with a split ratio of 10:1.

[0171] After GC-MS / MS analysis, the chromatogram was examined to calculate the peak area ratio. Compound identification was performed by comparing the mass spectrum data of the detected peaks with entries in the spectrum library. Peaks with significant area values ​​were selected, and the corresponding mass spectra were searched in the library to identify compounds with high similarity scores.

[0172] Table 1 shows the fatty acid composition in horse oil according to the barley koji fermentation period.

[0173] 고산 그리지 compare 1(HO) example 1 (HOF3) example 2 (HOF5) Example 3 (HOF10) Oleic acid amide 43.8 72.3 84.9 73.9 Palmitic amide 10.0 14.6 15.1 24.0 Stearic acid amide - - - 2.1 Palmitic acid (16:0) 20.4 5.2 - - Oleic acid (18:1) 11.9 - - - Squalene 6.5 2.3 - - Cholesterol 3.5 0.9 - - Myristic aldehyde 1.1 - - - 3-Hydroxypropyl oleate 1.2 - - - di-(9-octadecenoyl)-glycerol 1.5 - - - Ethyl palmitate - 1.4 - - Ethyl linoleate - 1.2 - - Vinyl Palmitate - 1.1 - - 1,2-Dipalmitoyl-sn-glycerol - 1.2 - -

[0174] Referring to Table 1, it was confirmed that the composition of major fatty acids in the fermented horse oil differs from that of horse oil depending on the fermentation period. It was confirmed that the fermented horse oil consists of oleic acid amide, palmitic acid amide, and stearic acid amide in a proportion of 86.9% to 100%, while horse oil consists of the above three fatty acids in a proportion of 53.8%.

[0175] Oleic acid amide was 72.3% in Example 1 (HOF3), which was on the 3rd day of koji fermentation, 84.9% in Example 2 (HOF5), which was on the 5th day of fermentation, and 73.9% in Example 3 (HOF10), which was on the 10th day of fermentation. Palmitic acid amide was 14.6% in Example 1 (HOF3), which was on the 3rd day of koji fermentation, 15.1% in Example 2 (HOF5), which was on the 5th day of fermentation, and 24.0% in Example 3 (HOF10), which was on the 10th day of fermentation, confirming that the proportion of palmitic acid amide increased as the koji fermentation period lengthened. Meanwhile, it was confirmed that stearic acid amide was 2.1% in Example 3 (HOF10), which was on the 10th day of fermentation.

[0176] Through the above results, it was confirmed that the major fatty acids of the fermented horse oil of the present invention are oleic acid amide, palmitic acid amide, and stearic acid amide, and it was found that there is a difference from the fatty acid composition of horse oil.

[0178] <Experimental Example 2> Molecular Docking Simulation Analysis

[0179] Molecular docking simulation analysis was performed on oleic acid amide, palmitic acid amide, and stearic acid amide, which are the major fatty acid components of fermented horse oil.

[0180] The 3D structure of the protein was determined using PyMOL 3.0.3 for docking simulation, and the molecular docking approach was analyzed according to the paper Hyun, KA et al. (2024, DOI: 10.3390 / ijms251910758). The target protein was human maltase-glucoamylase (MGAM) (PDB ID: 2QMJ). Ligands were provided from the PubChem database, and their 3D structures were downloaded for optimization. The ligands were optimized using the MMFF94 force field in OpenBabel, which included 3D coordinate generation and energy minimization to achieve the most stable form. AutoDock Tools 1.5.6 was used to add hydrogen atoms to the protein and ligands and to identify rotatable bonds in the ligands. The docking grid was defined based on the co-crystal ligand positions of the human maltase-glucoamylase (MGAM) structure, with center coordinates X, Y, Z of -20.4, -6.2, and -2.6, and the grid box size was 28.0 × 28.0 × 28.0. Docking was performed using the Lamarckian genetic algorithm, and molecular docking simulations were performed using AutoDock Vina 1.2.0.

[0181] Figure 1 shows the binding interactions between the human maltase-glucoamylase (MGAM) protein and its ligands. Figure 1(a) shows human maltase-glucoamylase, Figure 1(b) shows the binding interactions between MGAM and oleic acid amide, Figure 1(c) shows the binding interactions between MGAM and palmitate amide, and Figure 1(d) shows the binding interactions between MGAM and stearate amide.

[0182] Referring to Figure 1, it was confirmed that the binding affinities of oleic acid amide, palmitate amide, and stearate amide to MGAM were -5.2 kcal / mol, -5.1 kcal / mol, and -5.1 kcal / mol, respectively, and it was confirmed that all of the fatty acid amides form two hydrogen bonds through the amino group.

[0183] In the MGAM-oleic acid amide complex, it was confirmed that the amino group of oleic acid amide formed hydrogen bonds with ASP-542 (2.3 Å) and HIS-600 (2.5 Å), and the carbonyl group of the amide portion interacted with ARG526 through hydrogen bonds (3.4 Å), thus exhibiting slightly stronger bonding. In the case of the MGAM-palmitate amide complex, it was confirmed that the amino group of palmitate amide formed hydrogen bonds with ASP-542 (2.7 Å) and ASP-443 (2.5 Å). In the MGAM-stearic acid amide complex, it was confirmed that the amino group of stearic acid amide formed hydrogen bonds with ASP-542 (2.7 Å) and ASP-443 (2.5 Å).

[0184] Through the above results, it was found that the major fatty acids of the fermented horse oil of the present invention, namely oleic acid amide, palmitate amide, and stearate amide, exhibit molecular interactions with maltase-glucoamylase (MGAM) through hydrogen bonding, and thus the fermented horse oil of the present invention contributes to alpha-glucosidase inhibitory activity.

[0186] <Experimental Example 3> Analysis of Microbial Community Composition

[0187] Amplicon analysis was performed to analyze the microbial community composition of fermented horse oil. Changes in the microbial community of fermented horse oil fermented with barley koji were tracked using the 16S rRNA gene, a widely used marker in microbial community studies. To prepare 16S rRNA gene amplicons for sequencing, the Herculase II Fusion DNA Polymerase Nextera XT Index Kit V2 was used in conjunction with the MiSeq System (Illumina, USA).

[0188] DNA was extracted from the samples, and quality control (QC) was performed. The DNA samples were randomly fragmented, and 5' and 3' adapters were attached. The adapter-attached fragments were amplified via PCR and purified using gel electrophoresis to generate a sequencing library. Subsequently, microbial community analysis was performed using Illumina SBS (sequencing by synthesis) technology, and distinct clonal clusters were generated for accurate sequencing of the template.

[0189] Table 2 shows the changes in the composition of the microbial community and the relative abundance (%) of each microbial strain at three fermentation times (HOF3 (day 3), HOF5 (day 5), HOF10 (day 10)).

[0190] strain name division In the intestines of humans / animals Human / Animal Skin Habitat Example 1 (HOF3) (%) Example 2 (HOF5) (%) Example 3 HOF10 (%) Salmonella enterica pathogenic bacteria ○ - 19.4 14.8 0 Enterobacter quasihormaechei Opportunistic bacteria - - 24 20.2 1.40 Enterococcus faecalis Opportunistic bacteria ○ - 32.9 29.1 0.3 Lactobacillus acidophilus Beneficial bacteria ○ - 0 0 95.2 Atlantibacter hermannii Opportunistic bacteria - - 6.6 9.3 0 Enterococcus faecium Opportunistic bacteria ○ ○ 4.8 9.3 0 Enterococcus gallinarum Opportunistic bacteria ○ - 0.7 0 0 Enterococcus hirae Opportunistic bacteria ○ - 1.2 2.2 0 Enterococcus innesii Opportunistic bacteria ○ - 0 0.7 0 Enterobacter kobei Opportunistic bacteria - - <0.1 0 0 Staphylococcus warneri Opportunistic bacteria - ○ 1.5 1.4 0 Leuconostoc mesenteroides Opportunistic bacteria - - 0.4 0.4 0 Weissella paramesenteroides Opportunistic bacteria - - 1.4 1.8 0 Clostridium butyricum Opportunistic bacteria ○ - 0 <0.1 <0.1 Acetobacter persici Beneficial bacteria - - 0 0 0.5 Bacillus subtilis Beneficial bacteria ○ - 0 2.4 0.7 Bifidobacterium animalis Beneficial bacteria ○ - 0 0 0.5 Clostridium diolis Beneficial bacteria - - 0 4.4 2.6 Enterococcus mundtii Beneficial bacteria ○ - 1.5 1.3 0 Gluconobacter japonicus Beneficial bacteria - - <0.1 0 0 Lactococcus lactis Beneficial bacteria ○ - 0.2 0.3 0 Phytobacter palmae Beneficial bacteria - - 1.8 0 0 Pediococcus pentosaceus Beneficial bacteria ○ - 0.4 0.3 0 Lactococcus taiwanensis Beneficial bacteria 0.2 0.2 0 Clostridium beijerinckii Beneficial bacteria - - 0 0.3 0 Crassifilum sonorensis - - - <0.1 0.2 0 Other 0 1.5 0

[0191] Referring to Table 2 above, the microbial community on day 3 of fermentation (Example 1) was mainly Enterococcus faecalis ( Enterococcus faecalis )(32.9%), Salmonella enterica( Salmonella enterica )(19.4%), Enterobacter quacihormaetjei( Enterobacter quasihormaechei )(24.0%), Atlantibacter Hermannii( Atlantibacter hermanniIt was confirmed that it consisted of pathogenic and opportunistic bacteria, including )(6.6%). On the other hand, on day 10 of fermentation (Example 3), Enterobacter quacihormaechii ( Enterobacter quasihormaechei )(1.4%) and Enterococcus faecalis( Enterococcus faecalis )(0.3%) decreased to 1.4% and 0.3%, respectively, and Salmonella enterica( Salmonella enterica ) and Atlantibacter Hermani ( Atlantibacter hermanni ) was not detected, confirming that pathogenic and opportunistic bacteria were significantly reduced. In particular, on the 10th day of fermentation (Example 3), the beneficial bacterium Lactobacillus acidophilus ( Lactobacillus acidophilus It was confirmed that ) was the dominant species at 95.2%.

[0193] Table 3 shows the sum of the relative abundance (%) of the strains in Table 2 above according to pathogenicity.

[0194] division Example 1 (HOF3) (%) Example 2 (HOF5) (%) Example 3 HOF10 (%) pathogenic bacteria 19.4 14.8 0 Opportunistic bacteria <73.6 <74.5 <1.8 Beneficial bacteria <4.2 9.2 99.5 Other <0.1 1.7 0

[0195] Referring to Table 3 above, it was confirmed that as the fermentation period of horse oil with barley koji increased, the proportion of pathogenic bacteria decreased and the proportion of beneficial bacteria increased significantly.

[0196] Through the above results, it can be confirmed that the fermented horse oil of the present invention has an excellent effect of increasing the proportion of beneficial bacteria, and therefore, the fermented horse oil of the present invention can be usefully used as a food composition, pharmaceutical composition, quasi-drug composition, or feed composition for maintaining or improving the balance of intestinal microorganisms.

[0198] <Experimental Example 4> Anti-obesity and anti-diabetic efficacy

[0199] The anti-obesity and anti-diabetic efficacy was evaluated based on alpha-glucosidase inhibitory activity. For the examples and comparative examples, 750 mU / mL of alpha-glucosidase was incubated with 100 μL of 1.5 mM pNPG (p-nitrophenyl α-D-glucopyranoside) (Sigma-Aldrich, USA) in 100 mM sodium phosphate buffer (pH 6.8) at 37°C until the total reaction volume reached 200 μL. Acarbose (Sigma-Aldrich, USA), an alpha-glucosidase inhibitor, was used as a positive control. The reaction was initiated by adding pNPG to the sample and terminated after 10 minutes by adding 60 μL of 1 M Na2CO3. The release of pNP (p-nitrophenol) was expressed as a reading of absorbance measured at 405 nm using a spectrophotometer. To ensure accurate measurement, the background absorbance of the control group without α-glucosidase was excluded from the readings.

[0200] Figure 2 shows the alpha-glucosidase inhibitory activity. Figure 2(a) shows the alpha-glucosidase inhibitory activity of the control acarbose, and Figure 2(b) shows the alpha-glucosidase inhibitory activity of the comparative example (HO), example 1 (HOF3), example 2 (HOF5), and example 10 (HOF10).

[0201] Referring to Fig. 2(b), when the comparative example (HO) at a concentration of 12.5 μg / mL was treated with Example 1 (HOF3), a sample fermented on day 3, Example 2 (HOF5), a sample fermented on day 5, and Example 3 (HOF10), a sample fermented on day 10, alpha-glucosidase inhibition rates of 7.8%, 21.0%, 21.3%, and 37.5%, respectively, it was confirmed that the alpha-glucosidase inhibitory activity of the fermented horse oil of the present invention gradually increased with the fermentation period. In particular, it was confirmed that the alpha-glucosidase inhibitory activity of Example 3 (HOF10) was superior to that of the positive control, acarbose. While the positive control showed an inhibition rate of 28% at a concentration of 125 μg / mL, Example 3 showed an inhibition rate of 37% at a concentration of 12.5 μg / mL, confirming that it showed an effect more than 13.2 times superior.

[0202] Through the above results, it can be confirmed that the fermented horse oil of the present invention has excellent alpha-glucosidase inhibitory activity, so the fermented horse oil of the present invention can be usefully used as a food composition, pharmaceutical composition, quasi-drug composition, or cosmetic composition for the prevention, improvement, or treatment of body fat reduction and obesity, and can be usefully used as a food composition, pharmaceutical composition, or quasi-drug composition for the prevention, improvement, or treatment of diabetes.

[0204] <Experimental Example 5> Human Skin Irritation Test

[0205] The primary skin irritation tests of Examples 1 to 3 and Comparative Example 1 were conducted at a clinical trial institution specializing in dermatological evaluation (Dermapro Inc.) in accordance with the guidelines provided by the Ministry of Food and Drug Safety (MFDS), the Personal Care Products Council (PCPC), and the Standard Operating Procedures (SOP) of Dermapro Inc. Thirty-one female volunteers aged 20 to 60 years (mean age: 48.87 ± 5.60 years, range: 24 to 55 years) with no history of irritant or allergic contact dermatitis participated as subjects.

[0206] For the human skin primary irritation test, a sample concentration of 12.5 μg / mL prepared with a squalene-based formulation was used. The test product was applied to the backs of participants using the Van der Bend patch method and left undisturbed for 24 hours. After removing the patch, the skin condition was observed at 20 minutes and 24 hours after application. The results of the skin reaction at each test stage were calculated using the following Equation 1.

[0207]

[0208] Table 4 below shows the primary human irritation index for use as a cosmetic.

[0209] Range of Response Criteria 0.00 ≤ Response < 0.87 Slight 0.87 ≤ Response < 2.42 Mild 2.42 ≤ Response < 3.44 Moderate 3.44 ≤ Response Severe

[0210] Table 5 below shows the results of the human skin irritation test.

[0211] No division ( 12.5μg / mL ) participant 1st evaluation 2nd evaluation Reaction Rating (R) +1 +2 +3 +4 +1 +2 +3 +4 1 Comparative Example (HO) 0 0 0 0 0 0 0 0 0 0 2 Example 1 (HOF3) 0 0 0 0 0 0 0 0 0 0 3 Example 2 (HOF5) 0 0 0 0 0 0 0 0 0 0 4 Example 3 (HOF10) 0 0 0 0 0 0 0 0 0 0

[0212] * Hypoallergenic Category (None–Weak): 0.00 ≤ R < 0.87

[0213] Referring to Table 5 above, no skin reaction was observed in Examples 1 to 3 and the Comparative Example, and horse oil and fermented horse oil are judged to be substances in the non-irritating category in terms of primary irritation to human skin.

[0214] Through the above results, it was confirmed that the fermented horse oil of the present invention does not cause skin irritation when applied to cosmetic compositions and quasi-drug compositions.

[0215] Foregoing, specific parts of the present application have been described in detail. It is evident to those skilled in the art that such specific descriptions are merely preferred embodiments and that the scope of the present application is not limited thereto. Accordingly, the actual scope of the present application shall be defined by the appended claims and their equivalents. Furthermore, various modifications and improvements by those skilled in the art using the basic concept of the present application as defined in the claims are also within the scope of the rights of the present application.

Claims

Claim 1 A food composition for the prevention or improvement of diabetes or obesity, comprising a fermented horse oil extract as an active ingredient. Claim 2 A food composition for the prevention or improvement of diabetes or obesity, wherein the food composition of claim 1 has an alpha-glucosidase (α-glucosidase) inhibitory effect. Claim 3 Claim 1, wherein the food composition is a food composition for the prevention or improvement of diabetes or obesity that inhibits the absorption of sugar or fat. Claim 4 A food composition for the prevention or improvement of diabetes or obesity according to claim 1, wherein the fermented horse oil is obtained by mixing the horse oil with barley koji in a weight ratio of 5:1 to 1:5 and fermenting for 10 days or more. Claim 5 A food composition for the prevention or improvement of diabetes or obesity according to claim 1, wherein the fermented horse oil product changes the microbial community within the horse oil due to fermentation, and the change in the microbial community is one or more of the following: a decrease in the population of pathogenic bacteria or opportunistic bacteria; and an increase in the population of beneficial bacteria. Claim 6 In claim 5, the pathogenic bacteria or opportunistic bacteria are Salmonella enterica ( Salmonella enterica ), Enterobacter quacihormaechei( Enterobacter quasihormaechei ), Atlantibacter Hermani( Atlantibacter hermanni ), Enterococcus faecalis( Enterococcus faecalis ), Atlantibacter Hermani( Atlantibacter hermannii) , Enterococcus faecium( Enterococcus faecium ), Enterococcus galinarum( Enterococcus gallinarum ), Enterococcus hirae ( Enterococcus hirae ), Enterococcus inesi ( Enterococcus innesii ), Enterobacter covey ( Enterobacter kobei ) and Staphylococcus warneri Staphylococcus warneri A food composition for the prevention or improvement of diabetes or obesity, comprising one or more selected from the group consisting of ). Claim 7 In claim 5, the beneficial bacteria are Lactobacillus acidophilus ( Lactobacillus acidophilus ), Acetobacter persis ( Acetobacter persici ), Bacillus subtilis Bacillus subtilis ), Bifidobacterium animalis( Bifidobacterium animalis ), Clostridium diolis( Clostridium diolis ), Enterococcus mundti( Enterococcus mundtii ), Gluconobacter japonicus ( Gluconobacter japonicus ), Lactococcus lactis( Lactococcus lactis ) and Phytobacter palmae( Phytobacter palmae A food composition for the prevention or improvement of diabetes or obesity, comprising one or more selected from the group consisting of ). Claim 8 Claim 5, wherein the food composition is a food composition for the prevention or improvement of diabetes or obesity, which is intended for maintaining or improving the balance of intestinal microorganisms. Claim 9 A pharmaceutical composition for the prevention, improvement, or treatment of diabetes or obesity, comprising a fermented horse oil extract as an active ingredient. Claim 10 A pharmaceutical composition for the prevention, improvement, or treatment of diabetes or obesity, wherein the pharmaceutical composition of claim 9 has an alpha-glucosidase (α-glucosidase) inhibitory effect. Claim 11 In claim 9, the pharmaceutical composition is for maintaining or improving the balance of intestinal microorganisms, and is a pharmaceutical composition for the prevention, improvement, or treatment of diabetes or obesity. Claim 12 A quasi-drug composition for the prevention, improvement, or treatment of diabetes or obesity, comprising a fermented horse oil extract as an active ingredient and having an alpha-glucosidase inhibitory effect. Claim 13 In claim 12, the quasi-drug composition is for maintaining or improving the balance of intestinal microorganisms, and is a quasi-drug composition for the prevention, improvement, or treatment of diabetes or obesity. Claim 14 A cosmetic composition for the prevention or improvement of obesity, comprising a fermented horse oil extract as an active ingredient and having an alpha-glucosidase inhibitory effect. Claim 15 A feed composition for the prevention or improvement of diabetes or obesity, comprising a fermented horse oil product as an active ingredient and having an alpha-glucosidase inhibitory effect. Claim 16 Claim 15, wherein the feed composition is for maintaining or improving the balance of intestinal microorganisms, a feed composition for preventing or improving diabetes or obesity.

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