Herbal composition for promoting glp-1 secretion and inhibiting lipid accumulation

Abstract

The present disclosure relates to a functional composition having the effect of promoting GLP-1 secretion and inhibiting fat accumulation, and more specifically, to a functional composition effective in preventing or improving obesity and metabolic diseases using natural herbal medicines.

Description

Technology Field

[0001] The present disclosure relates to a functional composition having the effect of promoting GLP-1 secretion and inhibiting fat accumulation, and more specifically, to a functional composition effective in preventing or improving obesity and metabolic diseases using natural herbal medicines. Background Technology

[0002] In modern society, obesity is emerging as a serious global health issue. According to WHO statistics, the global obese population has more than tripled over the past 40 years, and it is reported that approximately 2.8 million people die annually due to obesity. Obesity is known as a major risk factor for various chronic diseases, including diabetes, hypertension, cardiovascular disease, and certain cancers, and the resulting socioeconomic burden is also increasing rapidly.

[0003] Various agents have been developed to address these obesity problems. In particular, interest in the enterocrine hormone GLP-1 (Glucagon-like peptide-1) has recently been increasing. GLP-1 is secreted by enterocrine cells upon food intake and plays an important role in energy metabolism by inducing satiety, promoting insulin secretion, and inhibiting glucagon secretion.

[0004] Currently available GLP-1 receptor agonists have been proven effective for weight loss, but they have limitations, such as being administered only in injectable form, frequent gastrointestinal side effects like nausea and vomiting, and the difficulty of long-term use due to high costs. Furthermore, recent studies have raised concerns that the long-term use of these synthetic drugs may increase the risk of conditions such as thyroid tumors.

[0005] Meanwhile, Korean traditional medicine has traditionally used various herbal medicines to treat obesity. Although several clinical studies have reported that specific herbal medicines are effective in weight loss and metabolic improvement, their mechanisms of action have not been clearly elucidated, and scientific verification from a modern medical perspective has been lacking. In particular, systematic research on the effects of herbal prescriptions on GLP-1 secretion or lipid metabolism has been very limited.

[0006] Furthermore, existing herbal obesity treatments lacked scientifically derived optimal blending ratios for medicinal herbs, and there was significant room for improvement in terms of the extraction and bioavailability of active ingredients. In addition, experimental verification regarding the effects of herbal compositions on inhibiting adipocyte differentiation and improving fatty liver was also insufficient.

[0007] Therefore, there is an urgent need for the development of a new, scientifically verified functional composition that can effectively promote GLP-1 secretion and inhibit fat accumulation. The problem to be solved

[0008] The present disclosure aims to solve the aforementioned problems by providing a new functional composition that utilizes natural herbal ingredients to effectively promote GLP-1 secretion and inhibit fat accumulation.

[0009] In addition, the present disclosure aims to provide an optimized extraction process that can maximize bioavailability while preserving the physiologically active substances of herbal medicines to the maximum extent.

[0010] In addition, the present disclosure aims to provide a functional composition that can effectively inhibit the differentiation of adipocytes, reduce fat accumulation in liver tissue, and improve blood lipid metabolism.

[0011] In addition, the present disclosure aims to provide a functional composition that is safe even with long-term use and has minimized side effects.

[0012] In addition, the present disclosure aims to provide a functional composition that can be manufactured into various formulations such as tablets, capsules, granules, and syrups, and has excellent storage stability.

[0013] In addition, the present disclosure aims to provide a functional food composition that can effectively inhibit the differentiation of adipocytes, reduce fat accumulation in liver tissue, and improve blood lipid metabolism.

[0014] In addition, the present disclosure aims to provide a functional food composition that is safe even with long-term use and has minimized side effects.

[0015] In addition, the present disclosure aims to provide a functional food composition that can be manufactured in various formulations such as tablets, capsules, granules, and syrups and has excellent storage stability.

[0016] The problems that the present disclosure aims to solve are not limited to those mentioned above, and problems that the present disclosure aims to solve that are not mentioned will be clearly understood by those skilled in the art from the description below. means of solving the problem

[0017] According to one aspect of the present disclosure, a functional composition comprising gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and ginger may be provided.

[0018] For example, a functional composition may be provided in which the gypsum content is 15-25 parts by weight based on 100 parts by weight of the total composition, and the Ziziphus jujuba content is 10-20 parts by weight based on 100 parts by weight of the total composition.

[0019] For example, a functional composition may be provided in which the content of apricot kernels is 10 to 20 parts by weight based on 100 parts by weight of the total composition, and the content of porridge kernels is 8 to 18 parts by weight based on 100 parts by weight of the total composition.

[0020] For example, a functional composition may be provided in which the content of gold is 5-15 parts by weight based on 100 parts by weight of the total composition, and the content of *Lycium chinense* is 5-15 parts by weight based on 100 parts by weight of the total composition.

[0021] For example, a functional composition may be provided in which the content of Ophiopogon japonicus is 8 to 18 parts by weight based on 100 parts by weight of the total composition, and the content of ginger is 3 to 13 parts by weight based on 100 parts by weight of the total composition.

[0022] For example, a composition in the form of a water extract, an ethanol extract, or a mixture thereof may be provided.

[0023] For example, a composition may be provided that is in the form of spherical granules, and the spherical granules have the characteristics of (i) an average particle size of 300-500 μm; (ii) a moisture content of 1-3 wt%; and (iii) a disintegration time of 15 minutes or less.

[0024] For example, a composition may be provided that is coated with an enteric coating agent, the enteric coating agent dissolves within 30 minutes in a phosphate buffer with a pH of 6.8, and the thickness of the enteric coating agent is 30-50 μm.

[0025] For example, a functional composition may be provided that is manufactured in the form of microspheres, wherein the microspheres are based on chitosan, alginic acid, or gelatin, and contains 0.5-1.5% by weight of glyceryl monostearate as a release retardant.

[0026] According to another aspect of the present disclosure, a functional food composition comprising gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and ginger may be provided.

[0027] For example, a functional food composition may be provided in which the gypsum content is 15-25 parts by weight based on 100 parts by weight of the total composition, and the Ziziphus jujuba content is 10-20 parts by weight based on 100 parts by weight of the total composition.

[0028] For example, a functional food composition may be provided in which the content of apricot kernels is 10 to 20 parts by weight based on 100 parts by weight of the total composition, and the content of ginseng is 8 to 18 parts by weight based on 100 parts by weight of the total composition.

[0029] For example, a functional food composition may be provided in which the content of goldenrod is 5-15 parts by weight based on 100 parts by weight of the total composition, and the content of Lycium chinense is 5-15 parts by weight based on 100 parts by weight of the total composition.

[0030] For example, a functional food composition may be provided in which the content of Ophiopogon japonicus is 8 to 18 parts by weight based on 100 parts by weight of the total composition, and the content of ginger is 3 to 13 parts by weight based on 100 parts by weight of the total composition.

[0031] For example, a functional food composition in the form of a water extract, an ethanol extract, or a mixture thereof may be provided.

[0032] For example, a functional food composition may be provided that is in the form of spherical granules, and the spherical granules have the characteristics of (i) an average particle size of 300-500 μm; (ii) a moisture content of 1-3 wt%; and (iii) a disintegration time of 15 minutes or less.

[0033] For example, a functional food composition may be provided that is coated with an enteric coating agent, the enteric coating agent dissolves within 30 minutes in a phosphate buffer with a pH of 6.8, and the thickness of the enteric coating agent is 30-50 μm.

[0034] For example, a functional food composition may be provided that is manufactured in the form of microspheres, wherein the microspheres are based on chitosan, alginic acid, or gelatin, and contains 0.5-1.5% by weight of glyceryl monostearate as a release retardant.

[0035] According to another aspect of the present disclosure, a method for preparing a functional composition for promoting GLP-1 secretion and inhibiting fat accumulation is provided, comprising the steps of: (a) enzymatically treating gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and dried ginger with fermented alcohol; (b) extracting the enzymatically treated herbal medicines with water; (c) ultrasonically treating the extracts; and (d) freeze-drying the ultrasonically treated extracts.

[0036] For example, a method for preparing a functional composition may be provided, characterized in that the enzyme treatment using fermented alcohol is performed at 40-50℃ for 12-24 hours.

[0037] For example, a method for preparing a functional composition may be provided, characterized in that water extraction is performed 2-3 times with water at 80-90°C.

[0038] For example, a method for manufacturing a functional composition may be provided, characterized in that the ultrasonic treatment is performed at a frequency of 40-50 kHz for 30-60 minutes.

[0039] According to another aspect of the present disclosure, a method for preparing a functional composition for promoting GLP-1 secretion and inhibiting fat accumulation is provided, comprising the steps of: (a) enzymatically treating gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and dried ginger with fermented alcohol; (b) extracting the enzymatically treated herbal medicines with water; (c) ultrasonically treating the extracts; and (d) freeze-drying the ultrasonically treated extracts.

[0040] For example, a method for preparing a functional food composition may be provided, characterized in that the enzyme treatment using fermented alcohol is performed at 40-50℃ for 12-24 hours.

[0041] For example, a method for preparing a functional food composition may be provided, characterized in that water extraction is performed 2-3 times with water at 80-90℃.

[0042] For example, a method for manufacturing a functional food composition may be provided, characterized in that the ultrasonic treatment is performed at a frequency of 40-50 kHz for 30-60 minutes. Effects of the invention

[0043] According to the present disclosure, a new functional composition that effectively controls appetite and inhibits fat accumulation by promoting GLP-1 secretion can be provided.

[0044] In addition, according to the present disclosure, a composition can be provided in which the bioactive substances of the herbal medicine are preserved to the maximum extent while the bioavailability is improved through an optimized extraction process.

[0045] In addition, according to the present disclosure, a functional composition that effectively inhibits the differentiation of adipocytes, reduces fat accumulation in liver tissue, and improves blood cholesterol and triglyceride levels may be provided.

[0046] In addition, according to the present disclosure, a functional composition that is non-cytotoxic, safe even with long-term use, and has minimized side effects can be provided.

[0047] In addition, according to the present disclosure, a functional composition that is easy to manufacture into various formulations and has excellent storage stability can be provided.

[0048] The superior and / or useful effects according to the present disclosure are not limited to the effects of the present disclosure described above, and those skilled in the art will also be able to readily recognize, based on the disclosure of the present specification, superior and / or useful effects of the present disclosure that are not explicitly disclosed in the present specification, and it should be understood that these are intentionally disclosed by the present specification and are obviously included within the scope of the present disclosure. Brief explanation of the drawing

[0049] FIG. 1 is a flowchart schematically illustrating a method for preparing a herbal functional composition for promoting GLP-1 secretion and inhibiting fat accumulation according to one embodiment of the present disclosure. Specific details for implementing the invention

[0050] Preferred embodiments of the present disclosure will be described below with reference to the attached drawings. However, embodiments of the present disclosure may be modified in various other forms, and the scope of the present disclosure is not limited to the embodiments described below.

[0051] According to one aspect of the present disclosure, a functional composition comprising gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and ginger may be provided.

[0052] According to one aspect of the present disclosure, a functional food composition comprising gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and ginger may be provided.

[0053] This composition can demonstrate an effect in improving obesity through two main mechanisms: promoting GLP-1 secretion and inhibiting fat accumulation. Here, gypsum can alleviate metabolic inflammation through its anti-inflammatory action, and Ziziphus jujuba can stabilize appetite by regulating gastrointestinal motility. Apricot kernel and Prunus persica seeds can enhance metabolic function through lipid metabolism improvement and the promotion of digestion and absorption, respectively, while Ophiopogon japonicus can maintain the function of GLP-1 secreting cells through its protective effect on the gastrointestinal mucosa. Scutellaria baicalensis can exhibit anti-inflammatory action, Lycium chinense bark can improve vascular endothelial cell function, and dried ginger can improve blood circulation.

[0054] Specifically, the combined action of these eight components can demonstrate obesity-improving effects through mechanisms such as the promotion of GLP-1 secretion from enterocrine L cells, activation of PKA C and AMPK signaling pathways, inhibition of adipocyte differentiation and reduction of fat accumulation, inhibition of fat accumulation in liver tissue, and improvement of blood lipid profiles. This combined action can produce synergistic effects that are difficult to achieve with a single component, and in particular, enables effective obesity improvement while minimizing side effects.

[0055] For example, a functional composition may be provided in which the gypsum content is 15-25 parts by weight based on 100 parts by weight of the total composition, and the Ziziphus jujuba content is 10-20 parts by weight based on 100 parts by weight of the total composition.

[0056] The mixing ratio of gypsum and Ziziphus jujuba may be closely related to the effect of promoting GLP-1 secretion. If the gypsum content is set to 15-25 parts by weight, metabolic inflammation can be effectively suppressed while preventing a decline in gastrointestinal function caused by excessive heat-clearing action. When Ziziphus jujuba is mixed at 10-20 parts by weight, a proper balance can be achieved between sedative effects and gastrointestinal motility regulation effects. This mixing ratio can optimize weight loss and appetite suppression effects while minimizing side effects.

[0057] Specifically, the interaction between gypsum and Ziziphus jujuba can activate PKA C and AMPK signaling pathways in enterocrine L cells. 15-25 parts by weight of gypsum can increase GLP-1 secretion in NCI-h716 cells by approximately 5.4 times, and 10-20 parts by weight of Ziziphus jujuba can sustain this secretion-promoting effect for more than 24 hours. In particular, with this formulation ratio, adipocyte differentiation can be inhibited by up to 46.4% without exhibiting cytotoxicity. This demonstrates a significantly improved effect compared to existing single-component formulations or other formulation ratios.

[0058] For example, a functional composition may be provided in which the content of apricot kernels is 10 to 20 parts by weight based on 100 parts by weight of the total composition, and the content of porridge kernels is 8 to 18 parts by weight based on 100 parts by weight of the total composition.

[0059] Adjusting the content of apricot kernels and burdock seeds can have a significant impact on lipid metabolism and digestive function. When the apricot kernel content is set to 10-20 parts by weight, the effects of inhibiting adipocyte differentiation and promoting lipolysis can be optimized. When burdock seeds are combined at 8-18 parts by weight, the effects of promoting gastrointestinal motility and improving digestion and absorption can be balanced. This composition ratio can significantly reduce the size of fat droplets in 3T3-L1 adipocytes.

[0060] Specifically, the combination of apricot kernels and Japanese apricot kernels can effectively inhibit the expression of SREBP1c and PPARγ, transcription factors related to adipocyte differentiation. In particular, this combination ratio reduces the expression of FABP4, which can inhibit adipocyte differentiation, while simultaneously promoting GLP-1 secretion in the gastrointestinal tract to exhibit an appetite-suppressing effect. This can exert a synergistic effect on weight loss and metabolic improvement.

[0061] For example, a functional composition may be provided in which the content of gold is 5-15 parts by weight based on 100 parts by weight of the total composition, and the content of *Lycium chinense* is 5-15 parts by weight based on 100 parts by weight of the total composition.

[0062] The combination ratio of Scutellaria root and Lycium chinense can play a key role in anti-inflammatory effects and the improvement of vascular endothelial cell function. Setting the Scutellaria root to 5-15 parts by weight can effectively suppress metabolic inflammation while exhibiting a liver-protective effect. When Lycium chinense is combined to 5-15 parts by weight, the improvement of vascular endothelial cell function and the regulation of lipid metabolism can be optimized. This combination ratio can reduce fat accumulation in liver tissue by approximately 26% in a high-fat diet-induced obesity model.

[0063] Specifically, the interaction between Scutellaria root and Lycium bark can regulate the expression of genes related to fat production in liver tissue. With this combination ratio, blood AST and ALT levels can be improved, while total cholesterol and triglyceride levels can be reduced simultaneously. In particular, as confirmed by H&E staining results of liver tissue, the size and number of fat droplets can be significantly reduced.

[0064] For example, a functional composition may be provided in which the content of Ophiopogon japonicus is 8 to 18 parts by weight based on 100 parts by weight of the total composition, and the content of ginger is 3 to 13 parts by weight based on 100 parts by weight of the total composition.

[0065] Adjusting the content of Ophiopogon japonicus and ginger can have a significant impact on gastrointestinal protection and blood circulation improvement. Setting the Ophiopogon japonicus content to 8-18 parts by weight can optimize the effects of protecting the gastrointestinal mucosa and maintaining the function of GLP-1 secreting cells. When ginger is formulated at 3-13 parts by weight, it can appropriately exhibit effects of improving blood circulation and promoting digestion and absorption. This formulation ratio ensures safety even with long-term use.

[0066] Specifically, the combination of Ophiopogon japonicus and Zingiber officinale can increase GLP-1 secretion in NCI-h716 cells in a concentration-dependent manner. As confirmed by experimental results, GLP-1 secretion can gradually increase without cytotoxicity and exhibit a secretion-promoting effect of approximately 5.4 times at the maximum concentration. In particular, at this combination ratio, the phosphorylation of PKA C and AMPK increases, which can sustain the GLP-1 secretion-promoting effect.

[0067] For example, a functional composition may be provided in the form of a water extract, an ethanol extract, or a mixture thereof.

[0068] The selection of extraction solvents can significantly influence the extraction efficiency and bioavailability of bioactive substances. Water extracts can be effective for extracting water-soluble components, while ethanol extracts can maximize the extraction of fat-soluble active components. Mixing the two types of extracts can enable the balanced extraction of various bioactive substances. This extraction method can optimize the bioavailability of GLP-1 secretion-promoting substances and fat accumulation inhibitors.

[0069] Specifically, the combination of enzymatic treatment with fermented alcohol and water extraction can significantly increase the effect of promoting GLP-1 secretion in NCI-h716 cells. As confirmed by experimental results, this extraction method can increase GLP-1 secretion in a concentration-dependent manner without cytotoxicity, and the effect can be sustained through the activation of PKA C and AMPK signaling pathways.

[0070] For example, a functional composition may be provided in the form of spherical granules, and the spherical granules may have the characteristics of (i) an average particle size of 300-500 μm; (ii) a moisture content of 1-3 weight%; and (iii) a disintegration time of within 15 minutes.

[0071] The physical properties of spherical granule formulations can have a decisive influence on the release and absorption of the drug. An average particle size of 300–500 μm can enable uniform dispersion and proper dissolution in the gastrointestinal tract. A moisture content of 1–3 wt% can induce proper disintegration while ensuring the stability of the formulation. A disintegration time of within 15 minutes can ensure rapid release and absorption of the drug.

[0072] Specifically, spherical granules with these physical properties can optimize the release of GLP-1 secretagogues in the gastrointestinal tract. As confirmed in animal studies, this formulation can effectively demonstrate weight loss and appetite suppression effects, and can reduce body weight by about 8%, particularly in a high-fat diet-induced obesity model.

[0073] For example, a composition may be provided in which a functional composition is coated with an enteric coating agent, the enteric coating agent dissolves within 30 minutes in a phosphate buffer with a pH of 6.8, and the thickness of the enteric coating agent is 30-50 μm.

[0074] Enteric coatings can prevent gastric degradation of the drug and enhance intestinal absorption. The characteristic of dissolving within 30 minutes in phosphate buffer at pH 6.8 can enable appropriate drug release in the small intestine. A coating thickness of 30–50 μm can optimize the balance between protection from gastric acid and intestinal release. These coating characteristics can improve the bioavailability of GLP-1 secretagogues.

[0075] Specifically, enteric-coated formulations can enable effective delivery of drugs to GLP-1 secreting cells in the small intestine. As confirmed by experimental results, these formulations can effectively promote intestinal GLP-1 secretion, which can lead to improved blood sugar control and appetite suppression effects.

[0076] For example, a functional composition may be provided in which the functional composition is manufactured in the form of microspheres, the microspheres are based on chitosan, alginic acid, or gelatin, and the functional composition contains 0.5-1.5% by weight of glyceryl monostearate as a release retardant.

[0077] Microsphere formulations can improve the sustained release and stability of drugs. Chitosan, alginate, and gelatin bases can provide appropriate drug release control characteristics along with biocompatibility. The addition of 0.5-1.5 wt% of glyceryl monostearate can optimize the drug release rate. These microsphere characteristics can enable a sustained effect of the drug.

[0078] Specifically, microsphere formulations can provide a sustained improvement effect through the sustained release of GLP-1 secretagogues. As observed in experimental results, these formulations can exhibit a GLP-1 secretagogue effect lasting for more than 24 hours, which suggests that effective improvement of obesity is possible with just one daily administration.

[0079] According to another aspect of the present disclosure, a functional food composition comprising gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and ginger may be provided.

[0080] For example, a functional food composition may be provided in which the gypsum content is 15-25 parts by weight based on 100 parts by weight of the total composition, and the Ziziphus jujuba content is 10-20 parts by weight based on 100 parts by weight of the total composition.

[0081] For example, a functional food composition may be provided in which the content of apricot kernels is 10 to 20 parts by weight based on 100 parts by weight of the total composition, and the content of porridge kernels is 8 to 18 parts by weight based on 100 parts by weight of the total composition.

[0082] For example, a functional food composition may be provided in which the content of gold is 5-15 parts by weight based on 100 parts by weight of the total composition, and the content of *Lycium chinense* is 5-15 parts by weight based on 100 parts by weight of the total composition.

[0083] For example, a functional food composition may be provided in which the content of Ophiopogon japonicus is 8 to 18 parts by weight based on 100 parts by weight of the total composition, and the content of ginger is 3 to 13 parts by weight based on 100 parts by weight of the total composition.

[0084] For example, a functional food composition in the form of a water extract, an ethanol extract, or a mixture thereof may be provided.

[0085] For example, a functional food composition may be provided that is in the form of spherical granules, and the spherical granules have the characteristics of (i) an average particle size of 300-500 μm; (ii) a moisture content of 1-3 weight%; and (iii) a disintegration time of 15 minutes or less.

[0086] For example, a functional food composition may be provided that is coated with an enteric coating agent, the enteric coating agent dissolves within 30 minutes in a phosphate buffer with a pH of 6.8, and the thickness of the enteric coating agent is 30-50 μm.

[0087] For example, a functional food composition may be provided that is manufactured in the form of microspheres, wherein the microspheres are based on chitosan, alginic acid, or gelatin, and contains 0.5-1.5% by weight of glyceryl monostearate as a release retardant.

[0088] According to another aspect of the present disclosure, a method for preparing a functional composition for promoting GLP-1 secretion and inhibiting fat accumulation is provided, comprising the steps of: (a) enzymatically treating gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and dried ginger with fermented alcohol; (b) extracting the enzymatically treated herbal medicines with water; (c) ultrasonically treating the extracts; and (d) freeze-drying the ultrasonically treated extracts.

[0089] Each of the four stages of the manufacturing process can serve a unique purpose and function. The enzymatic treatment step using fermented alcohol can effectively break down the structure of the cell walls of herbal medicines, thereby promoting the release of physiologically active substances. The subsequent water extraction step can maximize the extraction of water-soluble components through cell walls weakened by enzymatic treatment, and in particular, improve the extraction efficiency of polar compounds associated with GLP-1 secretion promotion. The ultrasonic treatment step can further increase the release of active ingredients by destroying remaining cell walls and reducing the size of microparticles through the cavitation effect. Finally, the freeze-drying step can prevent the denaturation of heat-sensitive physiologically active substances and ensure long-term storage stability.

[0090] Specifically, this stepwise manufacturing process can synergistically improve the extraction efficiency and stability of bioactive substances. As confirmed by experimental results, the composition prepared by this method can increase GLP-1 secretion in NCI-h716 cells in a concentration-dependent manner, showing an approximately 5.4-fold increase in secretion compared to the control group at the maximum concentration. Furthermore, it can reduce the size of lipid droplets in 3T3-L1 adipocytes by 46.4% and effectively suppress the expression of adipogenesis-related proteins such as FABP4, FAS, and SREBP1c. In particular, these effects can be sustained through the activation of PKA C and AMPK signaling pathways.

[0091] For example, a method for preparing a functional composition may be provided, characterized in that the enzyme treatment using fermented alcohol is performed at 40-50°C for 12-24 hours.

[0092] The temperature and time conditions of enzyme treatment using fermented alcohol can determine the extraction efficiency of physiologically active substances. A temperature range of 40-50℃ can optimize enzyme activity while preventing the decomposition of heat-sensitive components. A treatment time of 12-24 hours can ensure sufficient enzyme reaction while preventing excessive oxidation.

[0093] Specifically, under these enzyme treatment conditions, the cell wall structure is effectively degraded, which can increase the release of physiologically active substances. As observed in experiments, compositions treated under these conditions can maximize the effect of promoting GLP-1 secretion and can also effectively induce the activation of PKA C and AMPK signaling pathways.

[0094] For example, a method for preparing a functional composition may be provided, characterized in that water extraction is performed 2-3 times with water at 80-90°C.

[0095] The temperature and frequency of water extraction can play a key role in the extraction efficiency of water-soluble bioactive substances. An extraction temperature of 80–90°C can maximize the solubility of water-soluble components while minimizing the destruction of heat-sensitive components. Repeated extraction 2–3 times can maximize the recovery rate of active ingredients.

[0096] Specifically, under these water extraction conditions, GLP-1 secretion-promoting substances and fat accumulation-inhibiting substances can be effectively extracted. As confirmed by experimental results, compositions prepared under these conditions can effectively exhibit weight loss and appetite-suppressing effects, and can also significantly reduce fat accumulation in liver tissue.

[0097] For example, a method for manufacturing a functional composition may be provided, characterized in that the ultrasonic treatment is performed at a frequency of 40-50 kHz for 30-60 minutes.

[0098] The frequency and duration of ultrasonic treatment can have a decisive impact on the extraction efficiency and stability of bioactive substances. A frequency range of 40-50 kHz can effectively induce cell wall disruption while preventing the denaturation of bioactive substances. A treatment time of 30-60 minutes can ensure sufficient ultrasonic effects while suppressing excessive heat generation.

[0099] Specifically, under these ultrasonic treatment conditions, bioactive substances within the cell can be effectively eluted. As observed in experiments, compositions treated under these conditions can effectively inhibit adipocyte differentiation and significantly reduce the expression of transcription factors related to adipocyte differentiation, such as C / EBPα and PPARγ.

[0100] According to another aspect of the present disclosure, a method for preparing a functional composition for promoting GLP-1 secretion and inhibiting fat accumulation is provided, comprising the steps of: (a) enzymatically treating gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and dried ginger with fermented alcohol; (b) extracting the enzymatically treated herbal medicines with water; (c) ultrasonically treating the extracts; and (d) freeze-drying the ultrasonically treated extracts.

[0101] For example, a method for preparing a functional food composition may be provided, characterized in that the enzyme treatment using fermented alcohol is performed at 40-50℃ for 12-24 hours.

[0102] For example, a method for preparing a functional food composition may be provided, characterized in that water extraction is performed 2-3 times with water at 80-90°C.

[0103] For example, a method for preparing a functional food composition may be provided, characterized in that the ultrasonic treatment is performed at a frequency of 40-50 kHz for 30-60 minutes.

[0104] The present disclosure is described in detail below using specific embodiments and experimental examples. It should be understood that the following embodiments and experimental examples are not intended to limit the scope of the present disclosure, but are intended to illustrate the practice of the present disclosure.

[0105] Example 1: Preparation of a composition according to the present disclosure

[0106] The raw herbal ingredients gypsum, jujube seeds, apricot kernels, rhizomes, Ophiopogon japonicus, Scutellaria root, Lycium chinense bark, and dried ginger were prepared according to the Korean Pharmacopoeia standards. Each herbal ingredient was ground using a grinder (J-NCM, Jisico, Korea) to produce a powder that passes through a 40-mesh sieve. Each powdered herbal ingredient was precisely weighed in ratios of 20, 15, 15, 13, 13, 10, 10, and 8 parts by weight, and mixed using a V-mixer (VB-3, Tsutsui, Japan) for 30 minutes.

[0107] 10 L of fermented alcohol (30% ethanol) heated to 45°C was added to 1 kg of mixed raw herbal medicine, and 5 g each of alpha-amylase (Termamyl 120 L, Novozymes) and cellulase (Celluclast 1.5 L, Novozymes) were added and enzymatically treated with stirring at 45°C for 18 hours. The mixture after enzymatic treatment was filtered through a 5 μm pore filter (Whatman No. 5) to separate the solids.

[0108] 8 L of purified water heated to 85°C was added to the separated solids, and extraction was performed by stirring at 85°C for 2 hours. This process was repeated 3 times under the same conditions, and all the resulting extracts were combined. The combined extracts were treated using an ultrasonic processor (VC-750, Sonics) at 45 kHz and 500 W for 45 minutes. The treated extracts were frozen in a rapid freezer (SF-51, Samwon) at -40°C, and then dried using a vacuum freeze-dryer (PVTFD100R, Ilshinbiobase) at 20 mTorr and -40°C for 48 hours to obtain the final powder.

[0109] Example 2: Evaluation of physicochemical properties by formulation

[0110] 800g of the powder prepared in Example 1 was granulated using a fluid bed granulator (GPCG-1, Glatt) as follows. A 10% aqueous solution of hydroxypropylmethylcellulose (Pharmacoat 603, Shin-Etsu) was used as a binder, and granulation was performed under conditions of a spray rate of 8 mL / min, a spray pressure of 2.0 bar, an inlet air temperature of 60°C, and an exhaust temperature of 40°C.

[0111] Enteric coating was applied to the prepared granules using a fan coater (HCT-30, Erweka). The coating solution was prepared by dissolving 100 g of Oidragrit L100 (Evonik) and 10 g of triethyl citrate in 890 g of a mixed solvent of ethanol / acetone (6:4). Coating was performed under conditions of a spray rate of 3 mL / min, a spray pressure of 2.0 bar, a fan rotation speed of 12 rpm, and an exhaust temperature of 35°C. The coated granules were dried in a 40°C oven for 12 hours to obtain the final formulation.

[0112] Example 3: Cell-level efficacy evaluation

[0113] NCI-h716 cells were obtained from the Korean Cell Line Bank. Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin at 37°C under 5% CO2 conditions. 72 hours prior to the experiment, cells were seeded at a density of 1 x 10⁶ cells / well in 12-well plates coated with Matrigel (354234, Corning).

[0114] 3T3-L1 cells were purchased from the American Type Culture Collection. Cells were cultured in DMEM medium supplemented with 10% fetal bovine serum and seeded in 6-well plates at a density of 8 x 10⁴ cells / well. After the cells had fully grown, the medium was replaced with differentiation-inducing medium containing 10 μg / mL insulin, 1 μM dexamethasone, and 0.5 mM IBMX. Starting 2 days after differentiation, the medium was replaced with one containing only 10 μg / mL insulin and cultured for an additional 6 days.

[0115] Example 4: Evaluation of efficacy through animal experiments

[0116] 5-week-old male C57BL / 6J mice were acclimated to a standard diet for one week before being used in the experiment. The experimental groups were divided into a normal diet group (ND), a high-fat diet group (HFD), and a high-fat diet + composition administration group (LCM), with 8 mice assigned to each group. The normal diet group was fed a standard diet (10 kcal% fat), while the high-fat diet group and the composition administration group were fed a high-fat diet (60 kcal% fat) for 12 weeks. The composition administration group was orally administered the enteric-coated formulation prepared in Example 2 at a dose of 250 mg / kg once daily.

[0117] During the experiment, body weight was measured three times a week, and feed intake was measured daily. An oral glucose tolerance test was performed one week prior to the end of the experiment. After a 16-hour fast, glucose (2 g / kg) was administered orally, and blood glucose levels were measured by drawing blood from the tail vein at 0, 15, 30, 60, and 120 minutes. At the end of the experiment, blood was collected via cardiac angiography after a 16-hour fast, and liver tissue and epididymal adipose tissue were removed.

[0118] AST, ALT, total cholesterol, triglycerides, HDL-cholesterol, and LDL-cholesterol were measured in the blood. Liver and adipose tissues were fixed in 10% formalin and paraffin-embedded to prepare 4 μm thick sections. Histological changes were observed by hematoxylin-eosin staining, and the size and number of fat droplets were quantified using an image analysis program.

[0119] Experimental Example 1: Comparison of Efficacy of Compositions According to Manufacturing Conditions

[0120] Compositions were prepared according to the preparation method of Example 1 by varying the enzyme treatment temperature (40℃, 45℃, 50℃), water extraction temperature (80℃, 85℃, 90℃), and ultrasonic treatment frequency (40kHz, 45kHz, 50kHz). The content of active ingredients in the compositions prepared under each condition was quantified by HPLC, and biological activity was measured by the GLP-1 secretion amount in NCI-h716 cells and the inhibition rate of lipid accumulation in 3T3-L1 cells according to the method of Example 3. As a control, a composition prepared by water extraction only without fermented alcohol treatment was used. Table 1 shows the changes in the content of active ingredients and biological activity according to the enzyme treatment temperature, and Table 2 shows the changes in the content of active ingredients and biological activity according to the water extraction temperature.

[0121] temperature (℃) tBarbalin (mg / g) tBetaine (mg / g) tSaponin (mg / g) tGLP-1 (fold) tLipid inhibition(%) 40 t2.34±0.21 t1.87±0.15 t3.12±0.28 t3.8±0.4 t38.2±3.1 45 t3.65±0.32 t2.95±0.24 t4.56±0.41 t5.4±0.5 t46.4±3.8 50 t3.12±0.28 t2.43±0.19 t3.89±0.35 t4.2±0.4 t41.5±3.4 control group t1.23±0.11 t0.98±0.08 t1.65±0.15 t1.0±0.1 t12.3±1.1

[0122] temperature (℃) tBarbalin (mg / g) tBetaine (mg / g) tSaponin (mg / g) tGLP-1 (fold) tLipid inhibition (%) 80 t2.87±0.25 t2.34±0.19 t3.65±0.33 t4.2±0.4 t39.8±3.2 85 t3.65±0.32 t2.95±0.24 t4.56±0.41 t5.4±0.5 t46.4±3.8 90 t3.21±0.29 t2.56±0.21 t4.12±0.37 t4.8±0.4 t43.2±3.5 control group t1.23±0.11 t0.98±0.08 t1.65±0.15 t1.0±0.1 t12.3±1.1

[0123] Optimal effects were observed under conditions of an enzyme treatment temperature of 45℃, a water extraction temperature of 85℃, and an ultrasonic treatment frequency of 45kHz. Under these conditions, the content of baicalin, betaine, and steroid saponins was highest, GLP-1 secretion increased 5.4-fold compared to the control group, and triglyceride accumulation in adipocytes decreased by 46.4%. In particular, it was confirmed that the enzyme treatment temperature had the greatest influence on the physiological activity of the composition.

[0124] Experimental Example 2: Evaluation of Bioavailability of Enteric Coated Formulation

[0125] To evaluate the bioavailability and pharmacokinetic properties of the enteric-coated formulation prepared in Example 2, experiments were performed as follows.

[0126] Bioavailability was evaluated using 8-week-old male Sprague-Dawley rats (250–280 g, n=8 / group). Enteric-coated formulations and uncoated granules were administered orally at a dose of 250 mg / kg, and blood was collected from the jugular vein at 0, 0.5, 1, 2, 4, 8, 12, and 24 hours. Plasma was separated by centrifugation at 3,000 rpm for 10 minutes. The marker component (baicalin) in plasma was analyzed using LC-MS / MS (Agilent 6460), and naringin was used as the internal standard. Pharmacokinetic parameters were calculated using the Phoenix WinNonlin 8.1 program. The results are presented as follows.

[0127] 2-1. Pharmacokinetic parameters of enteric-coated formulations (mean ± standard deviation):

[0128] Cmax: 856±72 ng / mL

[0129] Tmax: 4.2±0.5 hr

[0130] AUC0-24hr: 5,234±486 ng·hr / mL

[0131] t1 / 2: 8.4±0.7 hr

[0132] Relative bioavailability: 243±22%

[0133] 2-2. Pharmacokinetic parameters of uncoated granules (mean ± standard deviation):

[0134] Cmax: 352±41 ng / mL

[0135] Tmax: 1.8±0.3 hr

[0136] AUC0-24hr: 2,156±234 ng·hr / mL

[0137] t1 / 2: 6.2±0.6 hr

[0138] Relative bioavailability: 100%

[0139] The enteric-coated formulation had a peak blood concentration 2.4 times higher than the uncoated granules, and bioavailability increased 2.4 times. In addition, it was confirmed that degradation in the upper gastrointestinal tract was effectively inhibited by delaying the time to reach peak blood concentration.

[0140] Experimental Example 3: Effect of promoting GLP-1 secretion and inhibiting fat accumulation

[0141] To evaluate the GLP-1 secretion-promoting effect and fat accumulation-inhibiting effect of the composition disclosed in the present disclosure using the cell experiment model of Example 3, an experiment was performed as follows.

[0142] NCI-h716 cells were treated with the composition of Example 1 at concentrations of 0, 62.5, 125, and 250 μg / mL for 2 hours, and the culture medium was collected to measure GLP-1 concentration using an ELISA kit (Millipore EGLP-35K). Phosphorylation of PKA C and AMPK was analyzed by western blot of cell lysates. Exendin-4 (100 nM), a GLP-1 secretagogue, was used as a positive control.

[0143] 3T3-L1 cells were treated with the composition at concentrations of 0, 62.5, 125, and 250 μg / mL in differentiation induction medium and cultured for 8 days. After Oil Red O staining, the cells were eluted with isopropanol, and absorbance was measured at 490 nm. The expression of lipogenesis-related proteins was analyzed by western blot. Orlistat (100 μM) was used as a positive control. The results are presented as follows.

[0144] 3-1. GLP-1 secretion (pg / mL) in NCI-h716 cells:

[0145] Control group: 78.0±0.7

[0146] 62.5μg / mL: 127.5±11.2

[0147] 125μg / mL: 333.9±29.8

[0148] 250μg / mL: 411.4±35.6

[0149] Exendin-4: 389.2±34.1

[0150] 3-2. Inhibition rate (%) of fat accumulation in 3T3-L1 cells:

[0151] Control group: 0

[0152] 62.5μg / mL: 28.4±2.5

[0153] 125μg / mL: 46.4±4.1

[0154] 250μg / mL: 52.7±4.6

[0155] Orlistat: 48.9±4.3

[0156] At a concentration of 250 μg / mL of the composition disclosed in this study, GLP-1 secretion increased 5.4-fold compared to the control group, which was similar to the level of Exendin-4. In addition, fat accumulation was inhibited by up to 52.7%, which was a superior effect compared to Orlistat.

[0157] Experimental Example 4: Weight loss effect in an obesity model

[0158] To evaluate the weight loss and metabolic improvement effects of the composition disclosed in the present disclosure using the animal model of Example 4, an experiment was performed as follows.

[0159] 5-week-old male C57BL / 6J mice were acclimatized for 1 week and then separated into groups as follows (n=8 / group):

[0160] - Normal Diet Group (ND): Standard feed (10 kcal% fat) + physiological saline

[0161] - High Fat Diet Group (HFD): High-fat feed (60 kcal% fat) + physiological saline

[0162] - Positive Control (PC): High-fat feed + Liraglutide (0.2 mg / kg / day)

[0163] - Test group (LCM): High-fat feed + Composition disclosed herein (250 mg / kg / day)

[0164] The test substance was administered orally once daily for 12 weeks while the animals were provided free-range. Body weight and feed intake were measured three times a week. An oral glucose tolerance test was performed prior to the end of the experiment, and blood and tissue samples were collected and analyzed. The results are presented as follows.

[0165] 4-1. Change in body weight (g) after 12 weeks:

[0166] ND: 31.2±2.1

[0167] HFD: 45.1±3.8

[0168] PC: 38.4±3.2

[0169] LCM: 41.5±3.3

[0170] 4-2. Cumulative feed intake (kcal / mouse / day):

[0171] ND: 5.3±0.4

[0172] HFD: 16.3±1.4

[0173] PC: 12.8±1.1

[0174] LCM: 12.4±1.0

[0175] 4-3. Improvement Rate (%) of Blood Lipid Indicators:

[0176] Total Cholesterol - PC: 18.2±1.6, LCM: 15.4±1.3

[0177] Triglycerides - PC: 20.5±1.8, LCM: 18.2±1.5

[0178] LDL-cholesterol - PC: 19.4±1.7, LCM: 16.8±1.4

[0179] 4-4. Reduction rate of fat content in liver tissue (%):

[0180] PC: 28.7±2.5

[0181] LCM: 26.4±2.3

[0182] The composition disclosed in this invention reduced body weight by 8.0% and feed intake by 23.9% compared to a high-fat diet after 12 weeks of administration. This was similar to the effect of Liraglutide (14.9% reduction in body weight and 21.5% reduction in feed intake).

[0183] Although embodiments of the present disclosure have been described in detail above, the scope of the present disclosure is not limited thereto, and it will be obvious to those skilled in the art that various modifications and variations are possible within the scope of the technical concept of the present disclosure as described in the claims.

Claims

Claim 1 A functional composition comprising gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and ginger. Claim 2 A functional composition according to claim 1, wherein the content of the gypsum is 15-25 parts by weight based on 100 parts by weight of the total composition, and the content of the Ziziphus jujuba is 10-20 parts by weight based on 100 parts by weight of the total composition. Claim 3 A functional composition according to claim 1 or 2, wherein the content of the apricot kernel is 10-20 parts by weight based on 100 parts by weight of the total composition, and the content of the porridge kernel is 8-18 parts by weight based on 100 parts by weight of the total composition. Claim 4 A functional composition according to any one of claims 1 to 3, wherein the content of gold is 5 to 15 parts by weight based on 100 parts by weight of the total composition, and the content of the root bark is 5 to 15 parts by weight based on 100 parts by weight of the total composition. Claim 5 A functional composition according to any one of claims 1 to 4, wherein the content of Ophiopogon japonicus is 8 to 18 parts by weight based on 100 parts by weight of the total composition, and the content of Ginger is 3 to 13 parts by weight based on 100 parts by weight of the total composition. Claim 6 A method for preparing a functional composition for promoting GLP-1 secretion and inhibiting fat accumulation, comprising: (a) enzymatically treating gypsum, Ziziphus jujuba, apricot kernel, ginseng, Ophiopogon japonicus, Scutellaria baicalensis, Lycium chinense bark, and dried ginger with fermented alcohol; (b) extracting the enzymatically treated medicinal materials with water; (c) ultrasonically treating the extract; and (d) freeze-drying the ultrasonically treated extract; characterized by comprising.

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