Symbiotic composition, metabolite, and use thereof for preventing and treating obesity-related diseases

Abstract

Provided are a symbiotic composition, a metabolite, and uses thereof for preventing and treating obesity-related diseases. A symbiotic composition comprising lychee polyphenols, Bifidobacterium longum, and methionine. Also provided is a symbiotic composition for preventing and treating obesity-related diseases. Also provided is a metabolite for preventing and treating obesity-related diseases.

Description

Technical Field

[0001] The present invention relates to compositions, metabolites and their uses, and particularly to synbiotic compositions, metabolites and their uses for preventing and treating obesity-related diseases.

Background Art

[0002] Since 1975, the global prevalence of obesity has almost doubled, mainly due to unhealthy eating habits. Since obesity is associated with an increased susceptibility to multiple chronic diseases, the spread of obesity has become an important global public health problem.

[0003] Diet patterns and the corresponding characteristics of the gut microbiota are associated with multiple health states. For example, a Western diet, which is usually high in fat and animal protein, increases the abundance of bacteria that produce trimethylamine (TMA) in the gut, and further increases the risk of cardiovascular disease and other health states.

[0004] Therefore, there is a need for improvement in the prior art regarding how to provide a composition for preventing and treating obesity-related diseases.

Summary of the Invention

Means for Solving the Problems

[0005] One embodiment of the present invention provides a synbiotic composition comprising lychee polyphenols, Bifidobacterium longum strains BCRC910812, BCRC12585, BCRC14602, BCRC11847, DSMZ20104, BCRC14607, BCRC11844, BCRC11846, BCRC14601, BCRC14606, BCRC14604, BCRC12584 or a combination thereof, and methionine.

[0006] In some embodiments, the lychee polyphenols include procyanidin, epicatechin (EC), or a combination thereof.

[0007] In some embodiments, based on 100% by weight of the total weight of the synbiotic composition, the weight percentage of the lychee polyphenols is 1% to 30%, the weight percentage of Bifidobacterium longum is 10% to 50%, and the weight percentage of methionine is 0.5% to 20%.

[0008] In some embodiments, Bifidobacterium longum is viable bacteria.

[0009] In some embodiments, the content of Bifidobacterium longum is 1×10 9 CFU / g to 1×10 11 CFU / g.

[0010] Another embodiment of the present invention provides a use of a synbiotic for manufacturing a composition for preventing and treating obesity-related diseases, wherein the synbiotic comprises lychee polyphenols, Bifidobacterium longum comprising the Bifidobacterium longum strains BCRC, BCRC12585, BCRC14602, BCRC11847, DSMZ20104, BCRC14607, BCRC11844, BCRC11846, BCRC14601, BCRC14606, BCRC14604, BCRC12584, or a combination thereof, and methionine.

[0011] In some embodiments, the lychee polyphenols include procyanidin, epicatechin, or a combination thereof.

[0012] In some embodiments, based on 100% by weight of the total weight of the symbiotic composition, the weight percentage of lychee polyphenols is 1% to 30%, the weight percentage of Bifidobacterium longum is 10% to 50%, and the weight percentage of methionine is 0.5% to 20%.

[0013] In some embodiments, Bifidobacterium longum is viable bacteria.

[0014] In some embodiments, obesity is diet-induced obesity.

[0015] In some embodiments, obesity-related diseases are selected from the group consisting of type 2 diabetes, hyperglycemia, glucose intolerance, dyslipidemia, insulin resistance, hyperinsulinemia, fatty liver, cardiovascular disease, stroke, cancer, and combinations thereof.

[0016] In some embodiments, the composition is a food composition or a pharmaceutical composition.

[0017] In some embodiments, the pharmaceutical composition is in an oral dosage form or a topical dosage form.

[0018] Another embodiment of the present invention is the use of a metabolite for producing a composition that suppresses fat accumulation, wherein the metabolite comprises 5'-methylthioadenosine, valine, pyroglutamic acid, glutamic acid, methionine, adenosine, 3-adenosine monophosphate, pyroglutamylvaline, or a combination thereof.

[0019] Another embodiment of the present invention is the use of a metabolite for producing a composition for preventing and treating obesity-related diseases, wherein the metabolite comprises 5'-methylthioadenosine, pyroglutamic acid, valine, glutamic acid, methionine, adenosine, 3-adenosine monophosphate, pyroglutamylvaline, or a combination thereof.

Brief Description of the Drawings

[0020] When reading the following detailed description with reference to the accompanying drawings, various aspects of the present invention will be most readily understood. It should be noted that various feature structures may not be drawn to scale in accordance with industry standard operating procedures. In fact, for clarity of discussion, the dimensions of various feature structures may be arbitrarily enlarged or reduced. The following description of the accompanying drawings is for the purpose of making the above and other objects, features, merits and embodiments of the present invention more clearly and readily understood.

Figure 1A - 1Q

Figure 2A - 2L

Figure 3A - 3K

Figure 4A - 4E

Figure 5A - 5B

Mode for Carrying Out the Invention

[0021] To further elaborate and enrich the description of the present invention, embodiments and specific examples of the present invention will be described below for illustrative purposes. However, this is not the only form for implementing or applying the specific examples of the present invention. Each of the examples disclosed below can be combined with or replaced by each other if beneficial, and other examples can be added to one example, and there is no need for further description or explanation. In the following description, many predetermined details are described in detail in order to enable the reader to fully understand the following examples. However, even without these predetermined details, the embodiments of the present invention can be implemented.

[0022] In this specification, unless otherwise specified in the text for the articles, "a" and "the" generally can refer to one or more. It should be further understood that the terms "comprising", "including", "having" and similar terms used in this specification are used to specify the described features, regions, integers, steps, operations, elements and / or components, but do not exclude the above or further one or more other features, regions, integers, steps, operations, elements, components, and / or groups thereof.

[0023] Also, when a number or range of numbers is described with terms such as "about", "approximate" and similar terms, the terms are intended to cover numbers within a reasonable range considering the variations, and these variations occur essentially during manufacturing as would be understood by those skilled in the art. As an example, a number or range of numbers includes a reasonable range that includes the described number, such as within + / - 10% of the described number, based on known manufacturing tolerances related to manufacturing features (having characteristics related to the number).

[0024] In this specification, the term "synbiotic" refers to a nutritional food or pharmaceutical that combines a prebiotic and a probiotic, and at the same time, a precursor of a postbiotic may be added to promote human health.

[0025] In this specification, the term "prebiotic" means a substance that can be decomposed and utilized by probiotics, promote the growth of probiotics, and bring about health-beneficial effects.

[0026] In this specification, the term "postbiotic" is an abiotic microbial secretion or its fragment product that can provide physiological benefits to the host.

[0027] In some specific embodiments of the present invention, the synbiotic composition is administered to an individual via an oral or parenteral route. In some specific embodiments of the present invention, the synbiotic composition is prepared in an oral dosage form selected from the group consisting of solutions, suspensions, emulsions, powders, tablets, pills, syrups, lozenges, tablets, chewable gums, and capsules and administered to an individual.

[0028] In some embodiments, the pharmaceutically acceptable carrier includes, but is not limited to, water, alcohols, glycol, preserving agents, antioxidants, solvent, emulsifier, suspending agent, decomposer, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant, absorption enhancers, active agents, humectants, odor absorbers, fragrances, pH adjusting agents, occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants, propellants, surfactant, and other similar or applicable carriers for the present invention.

[0029] In some embodiments of the present invention, the symbiotic composition may be a food composition. For example, it may be added to an edible material as a food additive for manufacturing food products that can be consumed by humans or animals. The food composition includes, but is not limited to, general foods, health foods, beverages, dietary supplements, dairy products, or feeds. In the example of an oral dosage form, the symbiotic composition may further selectively include carriers, excipients, and / or additives that are acceptable for pharmaceuticals and foods. In other examples, the dosage form of the complex probiotic composition may include, but is not limited to, powders, tablets, granules, suppositories, microcapsules, ampoules, agents for liquid spraying, or suppositories.

[0030] In some embodiments of the present invention, the synbiotic composition comprises lychee polyphenols, Bifidobacterium longum, and methionine.

[0031] In some embodiments, based on 100% by weight of the total weight of the synbiotic composition, the weight percentage of lychee polyphenols is 1% to 30%, the weight percentage of Bifidobacterium longum is 10% to 50%, and the weight percentage of methionine is 0.5% to 20%. The weight percentage of lychee polyphenols is 1% to 30%, for example, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or any value between any two of these values. The weight percentage of Bifidobacterium longum is 10% to 50%, for example, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or any value between any two of these values. The weight percentage of methionine is 0.5% to 20%, for example, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, or any value between any two of these values.

[0032] In some embodiments, the content of Bifidobacterium longum is 1×10 9 CFU / g to 1×10 11 CFU / g, for example, 2×10 9 CFU / g, 3×10 9 CFU / g, 4×10 9 CFU / g, 5×10 9 CFU / g, 6×10 9 CFU / g, 7×10 9 CFU / g, 8×10 9 CFU / g, 9×10 9 CFU / g, 1×10 10 CFU / g, 2×10 10 CFU / g, 3×10 10 CFU / g, 4×10 10 CFU / g, 5×1010 CFU / g, 6×10 10 CFU / g, 7×10 10 CFU / g, 8×10 10 CFU / g, 9×10 10 CFU / g, or any value between any two of these values.

[0033] In some embodiments of the present invention, there is provided a use of a metabolite for producing a composition for suppressing fat accumulation, wherein the metabolite comprises 5'-methylthioadenosine, valine, pyroglutamic acid, glutamic acid, methionine, adenosine, 3-adenosine monophosphate, pyroglutamylvaline, or a combination thereof.

[0034] In some embodiments of the present invention, there is provided a use of a metabolite for producing a composition for preventing and treating obesity-related diseases, wherein the metabolite comprises 5'-methylthioadenosine, valine, pyroglutamic acid, glutamic acid, methionine, adenosine, 3-adenosine monophosphate, pyroglutamylvaline, or a combination thereof.

[0035] Hereinafter, a plurality of examples and experimental examples are listed to explain the symbiotic composition of the present invention in more detail. However, these are merely for illustrative purposes and not for limiting the present invention. The protection scope of the present invention is based on the scope defined by the appended claims.

[0036] Example

[0037] Example 1 Lychee polyphenols can reduce HFD-induced obesity and improve intestinal microbiota dysbiosis

[0038] Litchi fruits (Litchi chinensis Sonn.) were obtained from a local market in Taipei, Taiwan, China. After manually removing the peel and seeds, the pulp was frozen in liquid nitrogen and dehydrated using a freeze dryer. Finally, the pulp was ground into a fine powder. Using an ultrasonic generator (30 W, 60 kHz), the dried litchi pulp powder (300 g) was extracted with a 70% aqueous methanol solution (2.4 L) for 30 minutes to obtain an extract. After filtering the extract through filter paper, the same extraction process was used to extract the residual compounds two more times to obtain a filtrate. The methanol in all the filtrates was removed using a rotary evaporator (45 °C). The polysaccharides in the filtrate were removed via an Oasis HLB column (20 cc / 1 g, Waters, Milford, MA, USA). The remaining compounds were centrifuged to obtain a crude extract. The litchi phenolic compounds in the crude extract were analyzed by the Global Natural Products Social (GNPS), which is a molecular networking method. The principle is that molecules with similar chemical structures exhibit similar MS / MS fragmentation patterns. The crude extract contains litchi polyphenols, which include proanthocyanidin (proanthocyanidin B2), epicatechin ((-)-epicatechin; EC), quercetin rhamnosyl-rutinoside, kaempferol rhamnosyl-rutinoside, isorhamnetin rhamnosyl-rutinoside, rutin, kaempferol rutinoside, and isorhamnetin rutinoside.

[0039] It has been discovered that lychee polyphenols can alleviate high-fat diet-induced obesity, reduce hyperlipidemia, and improve glucose tolerance and insulin sensitivity (Figures 1A - 1G). Application of lychee polyphenols enriched the diversity of the microbiota and improved the microbiota dysregulation caused by HFD (Figures 1H, 1I). Additionally, lychee polyphenols increased the abundance of the genus Bifidobacterium, with a slight increase in Bifidobacterium longum (Figures 1J, 1K).

[0040] Example 2 Bifidobacterium longum has anti-obesity activity

[0041] Subsequently, the regulatory ability of the metabolites of the genus Bifidobacterium on the host's metabolism was explored. For this purpose, all commercially available strains of the genus Bifidobacterium in Taiwan, China (11 strains, namely BCRC12585, BCRC14602, BCRC11847, DSMZ20104, BCRC14607, BCRC11844, BCRC11846, BCRC14601, BCRC14606, BCRC14604, BCRC12584, where BCRC is the Food Industry Research and Development Institute Bioresource Collection and Research Center in Taiwan, China, and DSMZ is the German Collection of Microorganisms and Cell Cultures) were purchased and then colonized (or called transcolonization) in C57BL / 6 germ-free mice. The germ-free mice were orally administered 1×10 9 colony-forming units (CFUs) of the genus Bifidobacterium (the content of 11 strains was mixed evenly) once a week with 0.2 mL of sterile PBS solution, and the control group of germ-free mice was given enteral nutrition with 0.2 mL of sterile PBS. All mice were fed a high-fat diet (OpenSource Diets TM, D12492, which has 60% of its calories derived from fat), were fed. To study whether the metabolites of the genus Bifidobacterium can reduce obesity, mouse feces (100 mg) were extracted with a 70% aqueous methanol solution (1 mL) containing cholic acid-d4 (2 ppm) as an internal standard. After homogenizing for 30 minutes using an ultrasonic generator (30 W, 60 kHz), the extract was centrifuged at 4 °C and 12,000 rpm for 10 minutes, and the supernatant was collected as the metabolite, and the metabolite was dried.

[0042] Measurement of lipid accumulation was performed. Briefly, to increase lipid accumulation, 70% proliferated HepG2 cells were cultured in RPMI 1640 medium containing 120 μg / mL of uric acid for 24 hours. After washing with PBS buffer, the cells were further maintained for 24 hours in fresh RPMI 1640 medium containing the metabolite (5.0, 1.0, 0.5, 0.1, and 0.05 μg / mL, the dried metabolite was redissolved in RPMI 1640 medium) and 120 μg / mL of uric acid. Cells maintained in pure RPMI 1640 medium were used as a control. According to the results, the metabolites extracted from the feces of mice fed with the genus Bifidobacterium were shown to significantly reduce lipid accumulation in a dose-dependent manner by testing the lipid accumulation activity of HepG2 cells (not shown). At the same time, the metabolites of each strain of the genus Bifidobacterium also exhibit anti-obesity activity (Figure 1L shows Bifidobacterium longum BCRC12585, and the remaining strains are not shown). According to these findings, it was shown that the anti-obesity effect of polyphenols may be due to the abundance of the genus Bifidobacterium. Since we attempt to identify specific chemical substances generated by probiotic microorganisms that have therapeutic potential and can be easily scaled up in the industry, Bifidobacterium longum subsp. - Bifidobacterium longum BCRC12585 (hereinafter referred to as Bifidobacterium longum) was selected. This is a commercially available strain isolated from humans and easily amplified, and is used to further explore its bioactive metabolites in a mouse model.

[0043] Next, until one week before death, 108 CFU of Bifidobacterium longum (BCRC12585) was administered once a day to C57BL / 6 mice. According to the results, the body weight of the mice decreased, and the weight gain decreased (Figs. 1M, 1N), and it was shown that the visceral fat and subcutaneous fat of the mice fed with Bifidobacterium longum slightly decreased (not shown). In addition, it was shown that the mice treated with Bifidobacterium longum had a significantly decreased fasting blood glucose (Figs. 1O, 1P, 1Q), and the glucose tolerance and insulin resistance were slightly improved (not shown).

[0044] Example 3 Identification of bioactive metabolites produced by Bifidobacterium longum

[0045] To further identify the bioactive metabolites produced by Bifidobacterium longum, a fractionation method was applied, followed by a combination of liquid chromatography-mass spectrometry (LC-MS) and an in vitro screening strategy (Figure 2A). Briefly, the extraction form of the metabolites of Bifidobacterium longum BCRC12585 (i.e., called Bifidobacterium longum metabolites or Bifidobacterium longum crude extract) was the same as in Example 2. Subsequently, 25 fractions were prepared from the Bifidobacterium longum metabolites through a pre-HPLC column, and then the anti-lipid accumulation activity of HepG2 cells was individually tested. It should be noted that fractions A to H with short retention times strongly inhibited lipid accumulation (Figure 2B), indicating that most of the active compounds are small molecules with high polarity. To identify candidate metabolites from each active component, further analysis was performed using a combination of liquid chromatography and high-resolution mass spectrometry (LC-MS / MS). Fourteen compounds, including nine amino acids (proline, valine, L-glutamine, glutamic acid (GA), arginine, pyroglutamic acid (PyroGA), methionine, isoleucine, and tryptophan), two peptides (cyclo-(His-Pro), pyro-glutamyl-valine (Pyro-glu-val)), adenosine, and two of its derivatives (3-adenosine monophosphate (3-AMPP) and 5'-methylthioadenosine (MTA)), were identified and further verified using reliable standards (not shown). Among these identified metabolites, eight metabolites (MTA, valine, pyroglutamic acid, glutamic acid, methionine, adenosine, 3-AMPP, and pyro-glutamyl-valine) significantly inhibited lipid accumulation in HepG2 cells (MTA is shown in Figure 2C, while the others are not shown in the figure). It should be noted that the abundance of MTA was negatively correlated with the biomarkers of obesity in most mice (Figure 2D). In summary, by integrating in vitro, in vivo, and metabolomics data, the bioactive metabolites that may be produced by Bifidobacterium longum were determined.

[0046] Interestingly, both MTA and its precursor methionine were identified in the active layer of Bifidobacterium longum and showed lipid-lowering effects in HepG2 cells (MTA is shown in Fig. 2C but not in the other figures). Next, it was investigated whether Bifidobacterium longum could produce MTA derived from methionine. As an experimental form, a high-fat diet (HFD) was administered for 17 weeks, and from the 8th week, MTA was administered by enteral nutrition at 100 mg per kilogram of body weight per day (100 mg / kg / day). As a precursor for MTA synthesis, methionine in the feces of mice treated with Bifidobacterium longum decreased, but MTA increased (Figs. 2E, 2F). Next, in vitro isotope labeling experiments were combined with mass spectrometry to understand whether Bifidobacterium longum could convert methionine to MTA. Briefly, Bifidobacterium longum was cultured on plates coated with 1 milliliter (mL) each of 0 mM, 1 mM, and 10 mM isotope-labeled methionine (Iso-met, -13C2H3) (Fig. 2G). According to the results, it was detected that 13C2H3-MTA with m / z 302.1173 (M+4) and its intermediate product 13C2H3-S-adenosyl-L-methionine (m / z 403.1657 (M+4)) had a dose-dependent effect (Figs. 2H-2K), indicating that Bifidobacterium longum could utilize methionine to synthesize MTA. Also, when methionine was administered, the content of MTA in the feces of mice increased (Fig. 2L). Additionally, it was discovered that other species of the genus Bifidobacterium, including Bifidobacterium breve, Bifidobacterium adolescentis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium bifidum, and Bifidobacterium animalis, used in this study, could also synthesize MTA using methionine (not shown). Here, a bioactive metabolite MTA with the potential for anti-obesity, which can be produced by the conversion of dietary amino acids by the genus Bifidobacterium, was discovered.

[0047] Example 4 MTA is a metabolite derived from methionine produced by Bifidobacterium longum and can improve obesity and metabolic disorders.

[0048] As a biosynthetic precursor of several important chemical substances, MTA plays a role in energy metabolism, including the regulation of the immune system, gene expression, and the synthesis of important biomolecules such as DNA and proteins. Therefore, next, the effect of MTA on host metabolism was studied by treating C57BL / 6 mice fed a high-fat diet (HFD) with or without MTA. As an experimental form, a high HFD was administered for 17 weeks, and from the 8th week, MTA was administered by enteral nutrition at 100 mg per kilogram of body weight per day (100 mg / kg / day). After administration of MTA, metabolic indices of mice, including body weight, weight gain during the study, visceral fat mass, and subcutaneous fat mass, were significantly decreased (Figures 3A - 3E). Similarly, it was observed that glucose tolerance and insulin sensitivity were also decreased (Figures 3F - 3I). In addition, liver weight and liver H&E staining showed that MTA could reduce hepatic steatosis in obese mice (Figures 3J, 3K). According to these results, it was shown that MTA can effectively regulate lipid metabolism and insulin sensitivity and reduce obesity and hepatic steatosis.

[0049] Example 5 MTA exerts multiple regulatory effects on liver energy metabolism.

[0050] To more comprehensively understand the potential of MTA as an anti-obesity molecule, liver RNA sequencing was performed on mice treated with MTA and Bifidobacterium longum to determine the potential mechanisms for preventing obesity. According to the first 30 differentially expressed molecular function gene ontology (GO) enrichment analyses, MTA has significant regulatory effects on genes related to fatty acid metabolism (Ltb4r1, Insig2, Elovl3, and Trib3), neurotransmitter signaling (Hcn3 and Adrb2), growth hormone signaling (Enho and Arntl), leptin signaling, and bile acid metabolism (not shown), while Bifidobacterium longum was shown to mainly regulate genes involved in fatty acid metabolism, bile acid metabolism, and immune response (not shown). Subsequently, the regulatory genes involved in these biological functions were deeply analyzed. It should be noted that in mice treated with MTA, the expression of insulin-induced gene 2 (Insig2), which affects liver cholesterol metabolism, lipogenesis, and glucose homeostasis, was decreased. Treatment with MTA also decreased fatty acid elongase 3 (Elovl3), an enzyme responsible for the elongation of very long-chain fatty acids. These findings indicated that the suppression of Insig2 and Elovl3 by MTA may contribute to fatty acid remodeling in the liver and to the regulation of body weight. In addition, the β-2-adrenergic receptor gene (Adrb2) and the leptin receptor gene (Lepr) were upregulated in mice treated with MTA, which promote metabolic signals related to energy consumption by adrenaline and leptin, respectively. Also, in mice treated with MTA, the expression of adropin (encoded by the energy homeostasis-related gene Enho) was increased, which may improve glucose metabolism by increasing glucose utilization. Similarly, overexpression of inhibin β-A encoded by the Inhba gene was able to improve glucose metabolism. It stimulated mitochondrial energy metabolism and increased energy consumption. These results indicated that MTA can improve energy metabolism by regulating the signaling of these metabolic hormones and proteins.In summary, according to these result tables, the preventive effect of MTA on obesity and insulin resistance may be due to multiple energy metabolism regulatory activities including fatty acid biosynthesis, glucose utilization, and energy metabolism, and these activities were shown to be potentially regulated by the regulation of the signal transduction of hormones and proteins with metabolic regulatory effects.

[0051] Bile acids have been considered as important signal transduction molecules that regulate fatty acids, cholesterol, energy, and glucose homeostasis. Therefore, enhancing bile acid synthesis could potentially be a strategy to avoid diet-induced obesity. Also, increased expression of Cyp7a1 (encoding the rate-limiting enzyme in the canonical bile acid synthesis pathway) was able to prevent high-fat diet-induced obesity, insulin resistance, and atherosclerosis. In view of the increased expression level of Cyp7a1 in mice treated with MTA (not shown), liver bile acid analysis was performed to determine whether the regulated Cyp7a1 expression level contributes to the bile acid composition. According to the results, the mice administered with MTA tended to have increased primary bile acids in the liver. In short, according to these results, it was shown that MTA can increase the expression level of Cyp7a1 and regulate bile acid synthesis and fatty acid metabolism downstream thereof.

[0052] Example 6 Suppressive effect on fat accumulation by the combination of lychee polyphenols, Bifidobacterium longum, and methionine

[0053] The synbiotic composition contained lychee polyphenols, Bifidobacterium longum, and methionine. Based on 100% by weight of the synbiotic composition, the lychee polyphenols in the synbiotic composition were purchased from commercially available OligonolR (including proanthocyanidins (proanthocyanidin A1, A2, B1, B2), catechin, epicatechin (EC), epicatechin gallate (ECG), epigallocatechin gallate (EGCG), epicatechin-(4β→8, 2β→O7)eicatechin-(4β→8)-eicatechin (A2-EC), and green tea polyphenols as the main active ingredients), with a content of 10 wt%. The Bifidobacterium longum was obtained from the B. Longum subsp. infantis BLI-02 strain (deposited at the Food Industry Research and Development Institute Biological Resources Conservation and Research Center BCRC910812 in Taiwan, China (this strain is freely distributable) or deposited at the China General Microbiological Culture Collection Center CGMCC No. 15212), with a content of 30 wt% and containing viable bacteria of 1×10 10 CFU / g (for example, 0.5 grams of the total weight of the synbiotic composition), and the content of methionine was 2 wt%. The synbiotic composition was manufactured as capsules, and the above weight percentages did not include the weight of the capsules.

[0054] Twenty-seven volunteers, eight males and nineteen females, met the screening criteria that the age of the subjects was between 18 and 65 years old. As an embodiment of this example, the subjects were made to take the symbiotic composition continuously for 5 weeks, and the administration method was 2 capsules once, twice a day. To control the variables, the dietitian provided the subjects with calorie and diet control according to their weight based on the calorie intake proposed by the body composition analysis results of the subjects. The subjects measured their body weight, body fat weight, body fat percentage, and visceral fat level with a body fat scale (InBody model number: InBody 570, a device that analyzes body composition using bioelectric impedance; BIA) 5 weeks before and after taking the symbiotic composition.

[0055] The results are shown in FIGS. 4A to 4E. As shown in FIG. 4A, compared with the body weight before taking, the body weight of the subjects decreased significantly after taking the symbiotic composition for 5 weeks, and the average body weight of the 27 people decreased by 1.39 kilograms. As shown in FIG. 4B, compared with the body fat weight before taking, the body fat weight of the subjects decreased significantly after taking the symbiotic composition for 5 weeks, and the average body fat of the 27 people decreased by 1.27 kilograms. As shown in FIG. 4C, compared with the body fat percentage before taking, the body fat percentage of the subjects decreased significantly after taking the symbiotic composition for 5 weeks, and the average body fat of the 27 people decreased by 1.21%. As shown in FIG. 4D, compared with the visceral fat level before taking, the visceral fat level of the subjects decreased significantly after taking the symbiotic composition for 5 weeks, and the average visceral fat level of the 27 people decreased by 0.7 level. As shown in FIG. 4E, compared with the abdominal circumference before taking, the abdominal circumference of the subjects decreased significantly after taking the symbiotic composition for 5 weeks, and the abdominal circumference of the 27 people decreased by an average of 2.57 centimeters.

[0056] In some embodiments, the lychee polyphenols in the synbiotic composition are obtained from Example 1, and the Bifidobacterium longum is obtained from BCRC12585, BCRC14602, BCRC11847, DSMZ20104, BCRC14607, BCRC11844, BCRC11846, BCRC14601, BCRC14606, BCRC14604, BCRC12584 or combinations thereof, and can also achieve the same effects of reducing body weight, reducing body fat weight, reducing body fat percentage, reducing visceral fat level, and reducing abdominal circumference at the waist.

[0057] Example 7 Inhibitory effect of MTA and pyroglutamic acid on fat accumulation

[0058] 3T3-L1 cells were seeded in a 12-well plate at a cell density of 3×10 3 cells / cm², and cultured in DMEM medium containing 10% bovine calf serum (CS), 100 U / mL penicillin and 100 μg / mL streptomycin for 3 to 4 days, and the medium was changed every 2 days until the cell density reached about 70% or more, that is, until differentiation. The medium was then replaced with DMEM medium containing 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 1 μM dexamethasone, 10 μg / mL insulin, 10% fetal bovine serum (FBS), 100 U / mL penicillin and 100 μg / mL streptomycin and cultured for 2 days. Subsequently, the medium was replaced with DMEM containing 10 μg / mL insulin, 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin to induce fat accumulation in 3T3-L1 cells. At the same time as induction, different compounds (MTA, pyroglutamic acid) were treated for 2 days.

[0059] As a result, as shown in Fig. 5A, even when 1 μg / mL or 5 μg / mL of MTA was administered, the accumulation of triglyceride (TG) in 3T3-L1 adipocytes could be significantly suppressed. As shown in Fig. 5B, when 10 μg / mL of MTA was administered, the accumulation of triglyceride in 3T3-L1 adipocytes was significantly suppressed, and there was also a tendency to suppress it when 2.5 μg / mL or 5 μg / mL of MTA was administered.

[0060] In some embodiments of the present invention, a molecular mechanism by which polyphenols can prevent obesity by enriching Bifidobacterium and MTA is disclosed. MTA is a bioactive metabolite of Bifidobacterium with anti-obesity activity. Also, polyphenols, Bifidobacterium, and methionine (a precursor of MTA) may be combined as a symbiotic product, and each component has anti-diabetic properties. The metabolic products MTA and pyroglutamic acid of Bifidobacterium longum have the effect of suppressing fat accumulation.

[0061] The present invention is disclosed as above in the embodiments, but this is not for limiting the present invention. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention is based on the scope defined by the appended claims.

Claims

1. Lychee polyphenols, Bifidobacterium longum strains BCRC910812, BCRC12585, BCRC14602, BCRC11847, DSMZ20104, BCRC14607, BCRC11844, BCRC11846, BCRC14601, BCRC14606, BCRC14604, BCRC12584 or a combination thereof, and methionine, A symbiotic composition comprising.

2. The symbiotic composition according to claim 1, wherein the lychee polyphenols comprise proanthocyanidins, epicatechin or a combination thereof.

3. Based on 100% by weight of the total weight of the symbiotic composition, the weight percentage of the lychee polyphenols is 1% to 30%, the weight percentage of the Bifidobacterium longum is 10% to 50%, and the weight percentage of the methionine is 0.5% to 20%. The symbiotic composition according to claim 1.

4. The symbiotic composition according to claim 1, wherein the Bifidobacterium longum is a viable bacterium.

5. The content of the Bifidobacterium longum is 1×10 9 CFU / g to 1×10 11 CFU / g. The symbiotic composition according to claim 4

6. Use of a symbiotic for the manufacture of a composition for preventing and treating obesity-related diseases, wherein the symbiotic comprises lychee polyphenols, Bifidobacterium longum strains BCRC, BCRC12585, BCRC14602, BCRC11847, DSMZ20104, BCRC14607, BCRC11844, BCRC11846, BCRC14601, BCRC14606, BCRC14604, BCRC12584 or a combination thereof, and methionine.

7. The use according to claim 6, wherein the lychee polyphenols comprise proanthocyanidins, epicatechin or a combination thereof.

8. Based on 100% by weight of the total weight of the symbiotic composition, the weight percentage of the lychee polyphenols is 1% to 30%, the weight percentage of the Bifidobacterium longum is 10% to 50%, and the weight percentage of the methionine is 0.5% to 20%. The use according to claim 6.

9. The use according to claim 6, wherein the Bifidobacterium longum is a viable bacterium.

10. The use according to claim 6, wherein the obesity is diet-induced obesity.

11. The use according to claim 6, wherein the obesity-related disease is selected from the group consisting of type 2 diabetes, hyperglycemia, impaired glucose tolerance, dyslipidemia, insulin resistance, hyperinsulinemia, fatty liver, cardiovascular disease, stroke, cancer, and combinations thereof.

12. The use according to claim 6, wherein the composition is a food composition or a pharmaceutical composition.

13. The use according to claim 12, wherein the pharmaceutical composition is in an oral administration or topical administration dosage form.

14. Use of a metabolite for the manufacture of a composition for preventing and treating obesity-related diseases, wherein the metabolite comprises 5'-methylthioadenosine, pyroglutamic acid, valine, glutamic acid, methionine, adenosine, 3-adenosine monophosphate, pyroglutamylvaline, or combinations thereof.

15. The use according to claim 14, wherein the obesity is diet-induced obesity.

16. The use according to claim 14, wherein the obesity-related disease is selected from the group consisting of type 2 diabetes, hyperglycemia, impaired glucose tolerance, dyslipidemia, insulin resistance, hyperinsulinemia, fatty liver, cardiovascular disease, stroke, cancer, and combinations thereof.

17. The use according to claim 14, wherein the composition is a food composition or a pharmaceutical composition.

18. The use according to claim 17, wherein the pharmaceutical composition is in an oral administration or topical administration dosage form.

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