Method and composition for regulating GLP-1

Abstract

Compositions and methods for improving or regulating GLP-1 levels, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion in a subject. The composition comprises two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin; or the composition comprises an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester; 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester; tetrahydrocurcumin, a physiologically acceptable salt, acid, ester; tributyrin; berberine; resveratrol; rhodiola rosea; cinnamon; fenugreek; olive leaf extract; grain of paradise; fruit-vegetable powder; whey protein; or arginine.

Description

Method and composition for regulating GLP-1BACKGROUND

[0001] Glucagon like peptide-1 (GLP-1), a peptide hormone from the intestinal tract, plays a central role in the coordination of postprandial glucose homeostasis through actions on insulin secretion, food intake and gut motility. Exogenous GLP-1 acutely stimulates insulin secretion, inhibits glucagon secretion, and lowers plasma glucose concentrations in people. In addition, GLP-1 inhibits gastric emptying, and reduces appetite in these patients. In rats, GLP-1 reverses age-dependent glucose decline in beta-cell function and stimulates beta-cell proliferation and neogenesis. In one report, after six weeks of GLP-1 treatment, 20 patients with type 2 diabetes in the study lost an average of 1.9 kg. Thus, GLP-1 could be effective on glycaemic control, body weight, insulin resistance, glucose tolerance management and beta-cell function.

[0002] In this invention, we have found that the composition can regulate GLP-1 levels for ameliorating metabolism.SUMMARY

[0003] In a first aspect, the present invention provides a method for improving or regulating GLP-1 level, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion in a subject, comprising administrating to the subject in need thereof a composition comprising two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin, or comprising administrating to the subject in need thereof a composition comprising: an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester,tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine.

[0004] In some embodiments, the method is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, the said managing glucose or insulin tolerance include stimulating insulin secretion, inhibiting glucagon secretion and reducing fasting glucose levels.

[0005] In some embodiments, the composition is administrating in an effective amount.

[0006] In some embodiments, the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletinranges from 1:40 to 40:1. In some embodiments, the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletinmay also be 1:40 to 40:1, 1:30 to 30:1, 1:20 to 20:1, 1:15 to 15:1, 1:10 to 10:1, 1:8 to 8:1, 1:5 to 5:1, 1:5 to 40:1, 1:3 to 40:1, 1:3 to 30:1, 1:2 to 20:1, 1:2 to 10:1, 1:1 to 10:1, 1:1 to 5:1, or 1:1. In some embodiments, the ratio of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine ranges from 1:200 to 200:1. In some embodiments, the ratio of the dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine may also be 1:100 to 200:1, 1:50 to 200:1, 1:50 to 100:1, 1:25 to 100:1, 1:20 to 100:1, 1:20 to 80:1, 1:15 to 80:1, 1:10 to 80:1, 1:8 to 80:1, 1:8 to 60:1, 1:5 to 60:1, 1:5 to 40:1, 1:3 to 40:1, 1:3 to 30:1, 1:2 to 20:1, 1:2 to 10:1, 1:1 to 10:1, 1:1 to 5:1, or 1:1.

[0007] In some embodiments, the composition is administrated in an amount of 1-3000 mg. In some embodiments, the composition is administrated in an amount of 1-2500 mg, 1-2000 mg, 1-1500 mg, 10-2000 mg, 10-1500 mg, 20-1000 mg, 10-800 mg, 20-600 mg, 30-300 mg, 40-200 mg, 40-100 mg. In some embodiments, the daily dose is administered in divided doses or a single dose. In some embodiments, the administration is at least once a day or more times a day. In some embodiments, the administration is at least 7 days and above in one period.

[0008] In some embodiments, the composition is administrated in an amount of 0.1 μM to 1 M or 0.01 to 50.0% (w / w). In some embodiments, the composition may be administrated in an amount of 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5 mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5 mM. In some embodiments, the composition may be administrated in an amount of 0.05% to 45%, 0.05% to 40%, 0.05% to 30%, 0.05% to 25%, 0.1% to 40%, 0.1% to 30%, 0.1% to 25%, 0.1% to 15%, 0. 5% to 30%, 0.5% to 20%, 0.5% to 10%, 1% to 35%, 1% to 25%, 1% to 10%, 2% to 25%, 2% to 10%, 5% to 9%, 1% to 7%, 2% to 7%, 0.05% to 7%, 0.05% to 9% (w / w).

[0009] In some embodiments, the subject is a mammal.

[0010] In some embodiments, the subject is human or animal. In some embodiments, the subject is human.

[0011] In some embodiments, the composition is prepared as a food, a drink, a supplement, a biochemical composition, or animal’s food, or a nutraceutical composition.

[0012] In some embodiments, the administration is through various routes selected from oral, intravenous injection, intramuscular injection, intraperitoneal injection, external use, or sublingual application.

[0013] In some embodiments, the composition is formulated in solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.

[0014] In a second aspect, the present invention provides a composition comprising two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin, or a composition comprising an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine.

[0015] In some embodiments, the composition is used for improving or regulating GLP-1 level, or stimulating GLP-1 secretion, or increasing GLP-1 receptors, or reducing GLP-1 degradation.

[0016] In some embodiments, the composition is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, the said managing glucose or insulin tolerance include stimulating insulin secretion, inhibiting glucagon secretion and reducing fasting glucose levels.

[0017] In some embodiments, the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletin ranges from 1:40 to 40:1. In some embodiments, the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletinmay also be 1:40 to 40:1, 1:30 to 30:1, 1:20 to 20:1, 1:15 to 15:1, 1:10 to 10:1, 1:8 to 8:1, 1:5 to 5:1, 1:5 to 40:1, 1:3 to 40:1, 1:3 to 30:1, 1:2 to 20:1, 1:2 to 10:1, 1:1 to 10:1, 1:1 to 5:1, or 1:1. In some embodiments, the ratio of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine ranges from 1:200 to 200:1. In some embodiments, the ratio of the dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine may also be 1:100 to 200:1, 1:50 to 200:1, 1:50 to 100:1, 1:25 to 100:1, 1:20 to 100:1, 1:20 to 80:1, 1:15 to 80:1, 1:10 to 80:1, 1:8 to 80:1, 1:8 to 60:1, 1:5 to 60:1, 1:5 to 40:1, 1:3 to 40:1, 1:3 to 30:1, 1:2 to 20:1, 1:2 to 10:1, 1:1 to 10:1, 1:1 to 5:1, or 1:1.

[0018] In some embodiments, the composition is administrated at a daily dose of 1-3000 mg. In some embodiments, the composition is administrated at a daily dose of 1-2500 mg, 1-2000 mg, 1-1500 mg, 10-2000 mg, 10-1500 mg, 20-1000 mg, 10-800 mg, 20-600 mg, 30-300 mg, 40-200 mg, 40-100 mg. In some embodiments, the daily dose is administered in divided doses or a single dose. In some embodiments, the administration is at least once a day or more times a day. In some embodiments, the administration is at least 7 days and above in one period.

[0019] In some embodiments, the composition is administrated in an amount of 0.1 μM to 1 M or 0.01 to 50.0% (w / w). In some embodiments, the composition may be administrated in an amount of 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5 mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5 mM. In some embodiments, the composition may be administrated in an amount of 0.05% to 45%, 0.05% to 40%, 0.05% to 30%, 0.05% to 25%, 0.1% to 40%, 0.1% to 30%, 0.1% to 25%, 0.1% to 15%, 0. 5% to 30%, 0.5% to 20%, 0.5% to 10%, 1% to 35%, 1% to 25%, 1% to 10%, 2% to 25%, 2% to 10%, 5% to 9%, 1% to 7%, 2% to 7%, 0.05% to 7%, 0.05% to 9% (w / w).

[0020] In some embodiments, the subject is a mammal.

[0021] In some embodiments, the subject is human or animal. In some embodiments, the subject is human.

[0022] In some embodiments, the composition is prepared as a food, a drink, a supplement, a biochemical composition, or animal’s food, or a nutraceutical composition.

[0023] In some embodiments, the composition is formulated in solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.

[0024] In a third aspect, the present invention provides use of composition in a preparation of a food, a drink, a supplement, a biochemical composition, animal’s food, or a nutraceutical composition for improving or regulating GLP-1 level, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion in a subject, wherein the composition comprises two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin, or the composition comprises an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine.

[0025] In some embodiments, the composition is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, the said managing glucose or insulin tolerance include stimulating insulin secretion, inhibiting glucagon secretion and reducing fasting glucose levels.

[0026] In some embodiments, the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletinranges from 1:40 to 40:1. In some embodiments, the ratio of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine ranges from 1:200 to 200:1.

[0027] In some embodiments, the composition is administrated at a daily dose of 1-3000 mg. In some embodiments, the composition is administrated at a daily dose of 1-2500 mg, 1-2000 mg, 1-1500 mg, 10-2000 mg, 10-1500 mg, 20-1000 mg, 10-800 mg, 20-600 mg, 30-300 mg, 40-200 mg, 40-100 mg. In some embodiments, the daily dose is administered in divided doses or a single dose. In some embodiments, the administration is at least once a day or more times a day. In some embodiments, the administration is at least 7 days and above in one period.

[0028] In some embodiments, the composition is administrated in an amount of 0.1 μM to 1 M or 0.01 to 50.0% (w / w). In some embodiments, the composition may be administrated in an amount of 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5 mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5 mM. In some embodiments, the composition may be administrated in an amount of 0.05% to 45%, 0.05% to 40%, 0.05% to 30%, 0.05% to 25%, 0.1% to 40%, 0.1% to 30%, 0.1% to 25%, 0.1% to 15%, 0. 5% to 30%, 0.5% to 20%, 0.5% to 10%, 1% to 35%, 1% to 25%, 1% to 10%, 2% to 25%, 2% to 10%, 5% to 9%, 1% to 7%, 2% to 7%, 0.05% to 7%, 0.05% to 9% (w / w).

[0029] In some embodiments, the subject is a mammal.

[0030] In some embodiments, the subject is human or animal. In some embodiments, the subject is human.

[0031] In some embodiments, the administration is through various routes selected from oral, intravenous injection, intramuscular injection, intraperitoneal injection, external use, or sublingual application.

[0032] In some embodiments, the composition is formulated in solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.

[0033] These and other features, aspects, and advantages of the present invention will become better understood with reference to the following description and appended claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Fig. 1 is body weight of groups 1-6 of Example 2.

[0035] Fig. 2 shows changes in the proportion of fat content in mice of groups 1-6 of Example 2.

[0036] Fig. 3 is area under the curve (AUC) of oral glucose tolerance test (OGTT) in mice of groups 1-6 of Example 2.

[0037] Fig. 4 is area under the curve (AUC) insulin intraperitoneal tolerance test (IPTT) in mice of groups 1-6 of Example 2.

[0038] Fig. 5 is the fat pad weight of groups 1-6 of Example 2.

[0039] Fig. 6 is the insulin levels in serum of groups 1-6 of Example 2.

[0040] Fig. 7 is the TC levels in serum of groups 1-6 of Example 2.

[0041] Fig. 8 is the TG levels in serum of groups 1-6 of Example 2.

[0042] Fig. 9 is the LDL levels in serum of groups 1-6 of Example 2.

[0043] Fig. 10 is the HDL levels in serum of groups 1-6 of Example 2.

[0044] Fig. 11 is the GLP-1 levels in serum of groups 1-6 of Example 2.

[0045] Fig. 12 is body weight of groups 1-5 of Example 3.

[0046] Fig. 13 shows changes in the proportion of fat content in mice of groups 1-5 of Example 3.

[0047] Fig. 14 is area under the curve (AUC) of oral glucose tolerance test (OGTT) in mice of groups 1-5 of Example 3.

[0048] Fig. 15 is area under the curve (AUC) insulin intraperitoneal tolerance test (IPTT) in mice of groups 1-5 of Example 3.

[0049] Fig. 16 is the fat pad weight of groups 1-5 of Example 3.

[0050] Fig. 17 is the insulin levels in serum of groups 1-5 of Example 3.

[0051] Fig. 18 is the TC levels in serum of groups 1-5 of Example 3.

[0052] Fig. 19 is the TG levels in serum of groups 1-5 of Example 3.

[0053] Fig. 20 is the LDL levels in serum of groups 1-5 of Example 3.

[0054] Fig. 21 is the HDL levels in serum of groups 1-5 of Example 3.

[0055] Fig. 22 is the GLP-1 levels in serum of groups 1-5 of Example 3.DETAILED DESCRIPTION

[0056] As used herein, the term “or” is meant to include both “and” and “or.” In other words, the term “or” may also be replaced with “and / or.”

[0057] As used herein, the singular forms “a,” “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0058] As used herein, the term “comprise” or “include” and their conjugations, refer to a situation wherein said terms are used in their non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. It also encompasses the more limiting verb ‘to consist essentially of’ and ‘to consist of’.

[0059] As used herein, the term "effective amount" refers to the amount required to achieve the effect as taught herein. The specific effective dose level for any particular subject will depend upon a variety of factors including the conditions being treated and the severity of the conditions; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration; the duration of the treatment; and like factors well known in the medical arts. For example, it is well known within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired effect and to gradually increase the dosage until the desired effect is achieved.

[0060] One of skill in the art recognizes that an amount may be considered “effective” even if the condition is not totally eradicated or prevented, but it or its symptoms and / or effects are improved or alleviated partially in the subject.

[0061] As used herein, the term “physiologically acceptable” means pharmaceutically, physiologically, alimentarily, and / or nutritionally acceptable, and refers to those compositions or combinations of agents, materials, or compositions, and / or their dosage forms, which are within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0062] As used herein, the term “mammal” or “subject” may be used interchangeably to refer to any animal to which the presently disclosed methods and compositions may be applied or administered. The animal may have an illness or other disease, but the animal does not need to be sick to benefit from the presently disclosed methods and compositions. As such any animal may apply the disclosed combinations, compositions or kits, or be a recipient of the disclosed methods. “Mammal” includes, without limitation, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, and, in particular, humans. Although the animal subject is preferably a human, the methods and compositions of the invention have application in veterinary medicine.

[0063] The dosage of the composition may range broadly, depending upon the desired effects and the indication. The dosage may be a single one or a series of two or more given in the course of one or more days, as is needed by the subject. In some embodiments, the compounds are administered for a period of time, for example for a week or more, or for months or years.

[0064] As used herein, the term "administration" refers to the process of delivering a disclosed composition or active ingredient to a subject. The compositions of the invention can be administered in a variety of ways, including orally, intragastrically, and parenterally (e.g., intravenous and intraarterial as well as other suitable parenteral routes), in external use, and the like.

[0065] As used herein, a “parenteral solution” refers to a solution that can be administered elsewhere in the body than the mouth and alimentary canal. It is not delivered via the intestinal tract. For example, parenteral solution can be delivered intravenously.

[0066] As used herein, a “tonic” refers to a medicinal substance taken to give a feeling of vigor or well-being.

[0067] As used herein, a “syrup” refers to a thick sticky liquid derived from a sugar-rich plant, for example, sugar cane, corn, and maple.

[0068] Multiple techniques of administering a composition exist in the art including, but not limited to, oral, rectal, topical, aerosol, injection and parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intranasal and intraocular injections.

[0069] “Intraperitoneal” as used here means within or administered through the peritoneum. The peritoneum is a thin, transparent membrane that lines the walls of the abdominal (peritoneal) cavity and contains / encloses the abdominal organs such as the stomach and intestines.

[0070] As used herein, “sublingual” refers to situated or applied under the tongue.

[0071] A “functionalized food composition” includes a food composition that has a potentially positive effect on health beyond basic nutrition.

[0072] Various embodiments of the present invention provide for methods for administrating the composition to a subject for effectively improving or regulating GLP-1 level, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion in a subject. For instance, the method is used for losing weight, managing glucose tolerance, or reducing fasting glucose levels. For example, an amount of approximately 10 mg to approximately 1000 mg of the composition may be administered to a human. The composition may be administrated in a variety of forms, such as solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.

[0073] In some implementations, one or more additives may be included in the composition, such as flavorings (e.g., natural and / or artificial), vitamins, minerals, binders, and / or any other appropriate additive. The additives may alter flavor, color, and / or texture. The additives may increase palatability and / or facilitate inclusion in a delivery vehicle (e.g., tablet, food product, beverage product such as a drink mix, etc.). In some implementations, the composition may be processed to increase bioavailability, solubility, palatability, and / or combination with other compounds.

[0074] Any titles or subheadings used herein are for organization purposes and should not be used to limit the scope of embodiments disclosed herein.

[0075] The substances used in the invention comprising two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin can be obtained through commercial purchased. The substances used in the invention comprising dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, polymer, ester, or 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, polymer, ester, or tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine can be obtained through commercial purchased.

[0076] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for.

[0077] All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.Example 1

[0078] Human NCI-H716 cells (colorectal adenocarcinoma cells) were grown in suspension at 37℃, 5% CO2. The culture medium was RPMI 1640 supplemented with 10% FBS, 2 mM L-glutamine, 100 IU / ml penicillin and 100 μg / ml streptomycin. Endocrine differentiation was induced by seeding cells in dishes coated with Matrigel, in high-glucose DMEM, 10% FBS, 2 mM L-glutamine, 100 IU / ml penicillin, and 100 μg / ml streptomycin. Two days before the experiments, 1.5 x 106cells were seeded in 12-well culture plates coated with Matrigel and containing highglucose DMEM, 10% FBS, 2 mM L-glutamine, 100 IU / ml penicillin and 100 μg / ml streptomycin. On the day of the experiment, medium was replaced by Krebs-Ringer bicarbonate buffer (KRB) buffer (128.8 mmol / l NaCl, 4.8 mmol / L KCl, 1.2 mmol / L KH2PO4, 1.2 mmol / l MgSO4, 2.5 mmol / l CaCl2, 5 mmol / l NaHCO3, and 10 mmol / l HEPES, pH 7.4) containing 0.2% BSA and different concentrations of DHB (0 mM, 2 mM, 5 mM or 10 mM) or tributyrin (0 mM, 2 mM, 5 mM or 10 mM) or DHB + tributyrin (0 mM + 0 mM, 1 mM + 1 mM, 2.5 mM + 2.5 mM or 5 mM + 5 mM). Following incubating at 37℃ for 2 h, the supernatants were collected with the addition of 50 μg / ml phenylmethylsulfonyl fluoride and stored at -80℃ for analysis. The cells were scraped off and sonicated in a homogenization buffer (1 M HCl containing 5% formic acid, 1% trifluoroacetic acid and 1% NaCl). GLP-1 in supernatant and cells was measured by a GLP-1 active ELISA kit, respectively, according to the manufacturer's protocol. Protein content of the cells was determined using the Bradford protein assay. The GLP-1 content was normalized for the total protein of the cells. A total of 3 x 106cells were seeded in 6-well culture plates coated with Matrigel and incubated for 24 h. Medium was replaced by serum-free medium containing 0.2% BSA without or with DHB (10 mM) or tributyrin (10 mM) or DHB + tributyrin (5 mM + 5 mM). The cells were re-incubated for 24 h and were washed with cold-PBS and stored at -80℃ for RT-PCR analysis. Total RNA was isolated from each well with Trizol reagent (Invitrogen Co., USA). RT-PCR was performed according to the protocol provided with the TwoStep RT-PCR kit (BestBio, Shanghai, China). The sequences of the forward and reverse primers (from 5' to 3') were: SEQ ID NO: 1 and SEQ ID NO: 2 for the proglucagon gene, SEQ ID NO: 3 and SEQ ID NO: 4 for prohormone convertase 3 gene, SEQ ID NO: 5 and SEQ ID NO: 6 for internal control GAPDH gene. After denaturing at 94℃ for 5 min, the amplification was obtained by 33 cycles of 94℃ for 30 s, 59℃ for 30 s and 72℃ for 30 s each. A final extension step at 72℃ for 5 min was performed. 27 cycles were performed for the amplification for GAPDH gene. PCR products were subjected to electrophoresis on 2.5% agarose gel, and visualized by means of ethidium bromide staining. Densitometric quantification was recorded using Jeda image analysis system 3.3 (Jiangsu Jeda ScienceTechnology Co. Ltd., Nanjing, China). Genes implicated in GLP-1 synthesis and secretion (i.e. Gcg, Pcsk1, and Slc5a1) were also detected by q-PCR.

[0079] Measurement of GLP-1: After incubation for 2 hours, GLP-1 content in supernatant and cell lysate was detected by ELISA.

[0080] mRNA expression of proglucagon and prohormone convertase 3: After incubation for 24 hours, mRNA of cells was extracted, cDNA was obtained by inversion, and the amplified fragments were amplified by PCR, and horizontal electrophoresis was performed to evaluate the relative expression levels of proglucagon mRNA and prohormone convertase 3 mRNA.

[0081] DHB or tributyrin or DHB + tributyrin treatment increase GLP-1 secretion (medium) in a concentration-dependent manner. DHB + tributyrin also increased cellular GLP-1 level, significant increases were found in cells treated with DHB or tributyrin alone.

[0082] Treatment with the compositions of the invention increase GLP-1 secretion (medium) in a concentration-dependent manner. The compositions also increase cellular GLP-1 level, significant increases are found.Example 2

[0083] At the beginning of the experiment, the mice were divided randomly into control group (Group 1, n=8), HFD group (Group 2, n=8), HFD + DHB + nobiletin group (Group 3, n=8), HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (Group 4, n=8), control + DHB + nobiletin group (Group 5, n=8), and control + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (Group 6, n=8). The dosage of DHB + nobiletin is 15 mg / kg + 100 mg / kg, and DHB + nobiletin + gymnema sylvestre + ceylon cinnamon is 15 mg / kg + 100 mg / kg + 130 mg / kg + 390 mg / kg. The group 1 is the control group and is given the normal chow; the group 2 is the high-fat mouse and continue high fat diet for 8 weeks; the groups 3 and 5 also continued to eat a high-fat diet and received daily intragastric administration of sample for 8 weeks; the groups 4 and 6 is given the normal chow and received daily intragastric administration of sample for 8 weeks. Record the body fat and weight of each group of mice every week. Complete Oral glucose tolerance test (OGTT) / Insulin intraperitoneal tolerance test (IPTT) on the last day of week 8, then execute the mice and take serum -80℃ for storage.

[0084] Measurement of Blood Glucose: Oral glucose tolerance test (OGTT): At week 16th of the experiment, after fasting for 15 h, Glucose was given intravenously (2 g / kg), and blood Glucose was measured at 0, 15, 30, 60, 90, 120 mins. Insulin intraperitoneal tolerance test (IPTT): At week 16th of the experiment, after fasting for 15 h, insulin (1U / kg) was injected intraperitoneally and blood glucose was measured at 0, 15, 30, 60, 90, 120 mins.

[0085] Body composition analysis: During the experiment, the fat mass and lean mass was measured by Nuclear Magnetic Resonance Body Composition Analyzer once a week.

[0086] Serum detection by ELISA: At week 8th of the experiment, the whole blood is extracted by removing the eyeball. The serum was acquired after centrifugation (10000 rpm 15 mins), stored at -80℃ for later use. Determinate the concentration of insulin (INS), cholesterol (TC), triglycerides (TG), LDL (low density lipoprotein), HDL (high density lipoprotein), GLP-1 in serum by ELISA kits.

[0087] Weight of the fat pad: At the 17th week, the mice were sacrificed, and the fat pads (subcutaneous, epididymis and perirenal) were dissected, photographed and weighed.

[0088] Change of body weight: Weigh once a week at a fixed time from beginning of the experiment.

[0089] Figure 1 is body weight of groups 1-6. After taking DHB + nobiletin for 8 weeks, body weight in HFD + DHB + nobiletin group (group 3) decreased by 4.0% compared with HFD group (group 2), NC + DHB + nobiletin group (group 5) decreased by 3.4% at the 8th week compared with NC group (group 1). After taking DHB + nobiletin + gymnema sylvestre + ceylon cinnamon for 8 weeks, body weight in HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 4) decreased by 7.1% compared with HFD group (group 2), NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) decreased by 5.8% at the 8th week compared with NC group (group 1). Thus, the body weight curve showed that DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon were helpful in inhibiting high fat induced obesity.

[0090] Figure 2 displays fat content percentage changes of groups 1-6. HFD + DHB + nobiletin (group 3) reduced fat accumulation by 7.64% as compared with HFD group (group 2), and the fat content in NC + DHB + nobiletin group (group 5) was decreased by 15.36% as compared to control mice (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) reduced fat accumulation by 17.80% and the fat content in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) was decreased approximately by 31.02% as compared to control mice (group 1).

[0091] Figure 3 is area under the curve (AUC) of oral glucose tolerance test (OGTT) of groups 1-6, reveals HFD + DHB + nobiletin (group 3) improved glucose tolerance by 9.8% as compared to HFD group (group 2), and NC + DHB + nobiletin (group 5) improved glucose tolerance by 5.6% as compared to NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) improved glucose tolerance by 15.1% as compared to HFD group (group 2), and NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 6) improved glucose tolerance by 10.4% as compared to NC group (group 1).

[0092] Figure 4 is area under the curve (AUC) insulin intraperitoneal tolerance test (IPTT) of groups 1-6, shows HFD + DHB + nobiletin (group 3) improved insulin sensitivity by 9.0% as compared to HFD group (group 2), and NC + DHB + nobiletin (group 5) improved glucose tolerance by 5.5% as compared to NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) enhanced insulin sensitivity by 18.1% as compared to HFD group (group 2), and NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 6) improved glucose tolerance by 13.7% as compared to NC group (group 1).

[0093] Figure 5 is the fat pad weight of groups 1-6, shows HFD + DHB + nobiletin (group 3) lowered pararenal fat weight by 8.7% as compared to HFD group (group 2), and NC + DHB + nobiletin (group 5) by 4.3% as compared to NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) lowered pararenal fat weight by 19.4% as compared to HFD group (group 2), and NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 6) by 20.6% as compared to NC group (group 1).

[0094] Figure 6-11 are serum lipid metabolism indexes including insulin (INS), cholesterol (TC), triglycerides (TG), LDL (low density lipoprotein), HDL (high density lipoprotein), GLP-1 levels in all groups.

[0095] Figure 6 is insulin (INS) levels in serum showing HFD + DHB + nobiletin (group 3) decreased INS by 14.7% relative to HFD group (group 2) and a 9.3% reduction in NC + DHB + nobiletin group (group 5) compared with NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) decreased INS by 29.4% relative to HFD group (group 2) and a 24.0% reduction in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) compared with NC group (group 1).

[0096] Figure 7 is cholesterol (TC) levels revealing HFD + DHB + nobiletin (group 3) reduced TC by 11.5% relative to HFD group (group 2) and a 12.9% reduction in NC + DHB + nobiletin group (group 5) compared with NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) reduced TC by 36.9% relative to HFD group (group 2) and a 33.6% reduction in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) compared with NC group (group 1).

[0097] Figure 8 is triglycerides (TG) levels demonstrating HFD + DHB + nobiletin (group 3) lowered TG by 11.31% relative to HFD group (group 2) and a 9.65% reduction in NC + DHB + nobiletin group (group 5) compared with NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) lowered TG by 20.24% relative to HFD group (group 2) and a 28.95% reduction in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) compared with NC group (group 1).

[0098] Figure 9 is LDL levels where HFD + DHB + nobiletin (group 3) reduced LDL by 12.5% relative to HFD group (group 2) and a 12.3% reduction in NC + DHB + nobiletin group (group 5) compared with NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) reduced LDL by 28.7% relative to HFD group (group 2) and a 26.2% reduction in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) compared with NC group (group 1).

[0099] Figure 10 is HDL levels indicating HFD + DHB + nobiletin (group 3) increased HDL by 10.5% compared with HFD group (group 2) and an 9.5% increase in NC + DHB + nobiletin group (group 5) compared with NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) increased HDL by 20.7% compared with HFD group (group 2) and an 26.4% increase in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) compared with NC group (group 1). In addition, as for the ratio of HDL / LDL, compared with the HFD group (group 2), HFD + DHB + nobiletin group (group 3) showed a 26.3% increase, and the NC + DHB + nobiletin group (group 5) showed a 24.8% increase compared with NC group (group 1). Compared with the HFD group (group 2), HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 4) showed a 69.2% increase, and the NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) showed a 71.1% increase compared with NC group (group 1).

[0100] Figure 11 is GLP-1 levels showing HFD + DHB + nobiletin (group 3) increased GLP-1 by 29.9% compared with HFD group (group 2) and an 27.7% increase in NC + DHB + nobiletin group (group 5) compared with NC group (group 1). HFD + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon (group 4) increased GLP-1 by 80.2% compared with HFD group (group 2) and an 56.4% increase in NC + DHB + nobiletin + gymnema sylvestre + ceylon cinnamon group (group 6) compared with NC group (group 1).

[0101] The results indicated that supplementing DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon can achieve the beneficial effects on metabolic health than without DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon supplementation.

[0102] DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon supplementation could significantly decrease the pararenal fat pad weight relative to mice without receiving DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon supplementation.Example 3

[0103] At the beginning of the experiment, the mice were divided randomly into control mice (Group 1, n=8), HFD group (Group 2, n=8), HFD + tributyrin (Group 3, n=8), HFD + DHB (Group 4, n=8) and HFD + DHB + tributyrin (Group 5, n=8). The dosage of tributyrin supplementation is 300 mg / kg, DHB is 15 mg / kg and DHB + tributyrin is 15 mg / kg + 300 mg / kg. The group 1 is the control group and is given the normal chow; the groups 2, 3, 4 and 5 are the high-fat-diet induced obese mice and continue high fat diet for 8 weeks; the groups 3, 4 and 5 also continued to eat a high-fat diet and received daily intragastric administration of sample for 8 weeks. Record the body fat mass and weight of each group of mice every week. Complete oral glucose tolerance test (OGTT) / insulin intraperitoneal tolerance test (IPTT) on the last day of week 8, then anesthesia the mice and take serum -80℃ for storage.

[0104] Measurement of Blood Glucose: Oral glucose tolerance test (OGTT): At week 16th of the experiment, after fasting for 15 h, glucose was given intravenously (2 g / kg), and blood glucose was measured at 0, 15, 30, 60, 90, 120 mins. Insulin intraperitoneal tolerance test (IPTT): At week 16th of the experiment, after fasting for 15 h, insulin (1 U / kg) was injected intraperitoneally and blood glucose was measured at 0, 15, 30, 60, 90, 120 mins.

[0105] Body composition analysis: During the experiment, the fat mass and lean mass was measured by Nuclear Magnetic Resonance Body Composition Analyzer once a week.

[0106] Serum detection by ELISA: At week 8th of the experiment, the whole blood is extracted from heart. The serum was acquired after centrifugation (3000 rpm 10 mins), stored at -80℃ for later use. Determinate the concentration of insulin (INS), cholesterol (TC), triglycerides (TG), LDL (low density lipoprotein), HDL (high density lipoprotein), GLP-1 in serum by ELISA kits.

[0107] Weight of the fat pad: At the 17th week, the mice were sacrificed, and the fat pads (subcutaneous, epididymis and perirenal) were dissected and weighed.

[0108] Change of body weight: Weigh once a week at a fixed time from beginning of the experiment.

[0109] Figure 12 is body weight of groups 1-5. Body weight in HFD + Tributyrin group (group 3) decreased by 3.4% at the 8th week compared with HFD group (group 2). HFD + DHB group (group 4) decreased by 3.7% compared with HFD group (group 2). HFD + DHB + Tributyrin group (group 5) decreased by 6.0% compared with HFD group (group 2). Thus, the body weight curve showed that Tributyrin or DHB or DHB+Tributyrin were helpful in inhibiting high fat induced obesity.

[0110] Figure 13 displays fat content percentage changes of groups 1-5. HFD + Tributyrin (group 3) reduced fat accumulation by 5.95%, and the fat content in HFD + DHB group (group 4) was decreased by 6.98% as compared to HFD group (group 2). HFD + DHB + Tributyrin (group 5) reduced fat accumulation by 13.03% as compared to HFD mice.

[0111] Figure 14 is area under the curve (AUC) of oral glucose tolerance test (OGTT) of groups 1-5, reveals HFD + Tributyrin (group 3) improved glucose tolerance by 5.9% as compared to HFD group (group 2), and HFD + DHB improved glucose tolerance by 7.4% as compared to HFD group (group 2). HFD + DHB + Tributyrin (group 5) improved glucose tolerance by 12.6% as compared to HFD group (group 2).

[0112] Figure 15 is area under the curve (AUC) insulin intraperitoneal tolerance test (IPTT) of groups 1-5, shows HFD + Tributyrin (group 3) improved insulin sensitivity by 6.6% as compared to HFD group (group 2), and HFD + DHB improved glucose tolerance by 9.9% as compared to HFD group (group 2). HFD + DHB + Tributyrin (group 5) enhanced insulin sensitivity by 12.0% as compared to HFD group (group 2).

[0113] Figure 16 is the pararenal fat weight of groups 1-5, shows HFD + Tributyrin (group 3) lowered pararenal fat weight by 7.5% as compared to HFD group (group 2), and HFD + DHB by 8.2% as compared to HFD group (group 2). HFD + DHB + Tributyrin (group 5) lowered pararenal fat weight by 11.7% as compared to HFD group (group 2).

[0114] Figure 17 is insulin (INS) levels in serum showing HFD + Tributyrin (group 3) decreased INS by 9.0% relative to HFD and a 12.1% reduction in HFD + DHB group (group 4) compared with HFD group (group 2). HFD + DHB + Tributyrin (group 5) decreased INS by 21.5% relative to HFD group (group 2).

[0115] Figure 18 is cholesterol (TC) levels revealing HFD + Tributyrin (group 3) reduced TC by 9.2% relative to HFD and a 10.6% reduction in HFD + DHB group (group 4) compared with HFD group (group 2). HFD + DHB + Tributyrin (group 5) reduced TC by 24.0% relative to HFD group (group 2).

[0116] Figure 19 is triglycerides (TG) levels demonstrating HFD + Tributyrin (group 3) lowered TG by 7.74% relative to HFD and a 10.12% reduction in HFD + DHB group (group 4) compared with HFD group (group 2). HFD + DHB + Tributyrin (group 5) lowered TG by 19.05% relative to HFD group (group 2).

[0117] Figure 20 is LDL levels where HFD + Tributyrin (group 3) reduced LDL by 8.1% relative to HFD and a 11.0% reduction in HFD + DHB group (group 4) compared with HFD group (group 2). HFD + DHB + Tributyrin (group 5) reduced LDL by 15.4% relative to HFD group (group 2).

[0118] Figure 21 is HDL levels indicating HFD + Tributyrin (group 3) increased HDL by 7.6% compared with HFD group (group 2) and an 9.1% increase in HFD + DHB group (group 4) compared with HFD group (group 2). HFD + DHB + Tributyrin (group 5) increased HDL by 13.8% compared with HFD group (group 2). In addition, as for the ratio of HDL / LDL, compared with the HFD group (group 2), HFD + Tributyrin group (group 3) showed a 17.1% increase, and the HFD + DHB group (group 4) showed a 22.6% increase compared with HFD group (group 2). HFD + DHB + Tributyrin group (group 5) showed a 34.5% increase compared with HFD group (group 2).

[0119] Figure 22 is GLP-1 levels showing HFD + Tributyrin (group 3) increased GLP-1 by 26.7% compared with HFD group (group 2) and an 28.3% increase in HFD + DHB group (group 4) compared with HFD group (group 2). HFD + DHB + Tributyrin (group 5) increased GLP-1 by 51.3% compared with HFD group (group 2).

[0120] The results indicated that supplementing DHB + tributyrin can achieve the beneficial effects on metabolic health than with DHB or tributyrin supplementation.

[0121] DHB + tributyrin supplementation could significantly decrease the pararenal fat pad weight relative to mice with DHB or tributyrin supplementation alone.

[0122] Insulin regulates glucose levels in the bloodstream and induces glucose storage in the liver, muscles, and adipose tissue. HDL have long been considered as "good cholesterol," beneficial to the whole body and, in particular, to cardio-vascular health. TG is a very important component of blood fat, and elevated levels may lead to heart disease. LDL, the "bad cholesterol" has a strong effect on atherosclerosis. Therefore, LDL is also known as atherogenic factor. When LDL, especially oxidized low-density lipoprotein (OX-LDL) is excessive, the cholesterol it carries accumulates in the artery wall, which is easy to cause arteriosclerosis for a long time. The importance of GLP-1 was described above. From the point of view of serum indexes, DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon supplementation could significantly increase HDL and GLP-1, and decrease INS, TC, TG, and LDL relative to mice without receiving DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon supplementation, indicating that DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon has the effect of improving obesity and ameliorating metabolism. Non-obesity mice supplemented with DHB + nobiletin or DHB + nobiletin + gymnema sylvestre + ceylon cinnamon decreased body weight and increased GLP-1 level without significantly differences compared with control group. From the point of view of serum indexes, DHB + tributyrin supplementation could significantly increase GLP-1 relative to mice with receiving DHB or tributyrin supplementation, indicating that DHB + tributyrin has the effect of improving obesity and ameliorating metabolism. This effect persists for as long as the test protocol is continued.

[0123] Although specific embodiments and examples of this invention have been illustrated herein, it will be appreciated by those skilled in the art that any modifications and variations can be made without departing from the spirit of the invention. The examples and illustrations above are not intended to limit the scope of this invention. Any combination of embodiments of this invention, along with any obvious their extension or analogs, are within the scope of this invention. Further, it is intended that this invention encompass any arrangement, which is calculated to achieve that same purpose, and all such variations and modifications as fall within the scope of the appended claims.

Claims

1. A method for improving or regulating GLP-1 level, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion in a subject, comprising administrating to the subject in need thereof a composition comprising two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin, or comprising administrating to the subject in need thereof a composition comprising: an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester,tributyrin, berberine, resveratrol,rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine.

2. The method of claim 1, wherein the method is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, the said managing glucose or insulin tolerance include stimulating insulin secretion, inhibiting glucagon secretion and reducing fasting glucose levels.

3. The method of claim 1 or 2, wherein the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletin ranges from 1:40 to 40:1, or the ratio of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine ranges from 1:200 to 200:1.

4. The method of any of claims 1-3, wherein the composition is administrated in an amount of 1-3000 mg, or 0.1 μM to 1 M or 0.01 to 50.0% (w / w).

5. The method of any of claims 1-4, wherein the subject is a mammal.6.The method of any of claims 1-5, wherein the subject is human or animal.7.The method of any of claims 1-6, wherein the composition is prepared as a food, a drink, a supplement, a biochemical composition, animal’s food, or a nutraceutical composition.8.The method of any of claims 1-7, wherein the administration is through various routes selected from oral, intravenous injection, intramuscular injection, intraperitoneal injection, external use, or sublingual application.9.The method of any of claims 1-8, wherein the composition is formulated in solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.10.A composition comprising two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof, gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon;nobiletin, or a composition comprising an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid,and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine.

11. The composition of claim 10, wherein the composition is used for improving or regulating GLP-1 level, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion.12.The composition of claim 10 or 11, wherein the composition is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, the said managing glucose or insulin tolerance include stimulating insulin secretion, inhibiting glucagon secretion and reducing fasting glucose levels.13.The composition of any of claims 10-12, wherein the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon, nobiletinranges from 1:40 to 40:1, or the ratio of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine ranges from 1:200 to 200:1.14.The composition of any of claims 10-13, wherein the composition is administrated at a daily dose of 1-3000 mg, or 0.1 μM to 1 M or 0.01 to 50.0% (w / w).15.The composition of any of claims 10-14, wherein the composition is formulated in solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.16.Use of composition in a preparation of a food, a drink, a supplement, a biochemical composition, animal’s food, or a nutraceutical composition for improving or regulating GLP-1 level, or increasing GLP-1 receptors, or reducing GLP-1 degradation, or stimulating GLP-1 secretion in a subject, wherein the composition comprises two or more of the following substances: an effective amount of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid; gymnema sylvestre or gymnemic acid; cinnamon or ceylon cinnamon; nobiletin, or the composition comprises an effective amount of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid, and one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine.17.The use of claim 16, wherein the composition is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, the said managing glucose or insulin tolerance include stimulating insulin secretion, inhibiting glucagon secretion and reducing fasting glucose levels.18.The use of claim 16 or 17, wherein the subject is a mammal.19.The use of any of claims 16-18, wherein the ratio of dihydroberberine (DHB) or berberine, a physiologically acceptable salt, polymer, ester, acid thereof to one or two of the following substances: gymnema sylvestre or gymnemic acid, cinnamon or ceylon cinnamon,nobiletinranges from 1:40 to 40:1, or the ratio of dihydroberberine (DHB), a physiologically acceptable salt, polymer, ester, acid to one or more of the following substances: β-aminoisobutyric acid, a physiologically acceptable salt, ester, 3-hydroxy-3-methylbutanoic acid, a physiologically acceptable salt, ester, tetrahydrocurcumin, a physiologically acceptable salt, acid, ester, tributyrin, berberine, resveratrol, rhodiola rosea, cinnamon, fenugreek, olive leaf extract, grain of paradise, fruit-vegetable powder, whey protein, or arginine ranges from 1:200 to 200:1.20.The use of any of claims 16-19, wherein the composition is administrated at a daily dose of 1-3000 mg or in an amount of 0.1 μM to 1 M or 0.01 to 50.0% (w / w).21.The use of any of claims 16-20, wherein the composition is formulated in solutions, aqueous suspensions, liquid suspensions, parenteral solutions, injections, microemulsion, (micro)capsules, drops, granules, liquids, powders, aerosols, tonics, syrups, tablets, pills, film, functionalized foods, beverages, toothpaste, nourishments, snacks, gums, bars, sugars, and sublingual articles.

Images