Use of dihydroberberine or its derivatives for regulating GLP-1

Abstract

Provided herein are novel compositions and methods for improving or regulating GLP-1 levels, or stimulating GLP-1 secretion in a subject. The composition may comprise dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof.

Description

BACKGROUND

[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 agedependent 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] The effect of dihydroberberine on GLP-1 has not been reported yet. In this invention, dihydroberberine (DHB) has been found to regulate GLP-1 levels. SUMMARY

[0003] In a first aspect, the present invention provides a method for improving or regulating GLP-1 level, or stimulating GLP-1 secretion in a subject, comprising administrating to the subject a composition comprising: an effective amount of dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof.

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

[0005] In some embodiments, dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative is administrating in an effective amount.

[0006] In some embodiments, compared with the untreated control group, the GLP-1 of DHB treated group was at least increased by 10%, preferably increased by 15%, 20%, 30%, 50%, 70%', 80%, 90%, 100%.

[0007] In some embodiments, the said ameliorating blood fat is the increased ratio of HDL / LDL.

[0008] In some embodiments, compared with the untreated control group, the body weight of DHB treated group was decreased approximately 5%, preferably decreased 7%, 9%; the fat content of DHB treated group reduced by approximately 15%, preferably decreased 19%, 30%, or 35%'; and the AUG of OGTT of DHB supplemented decreased by about 5%-, preferably decreased 10%', 15%, or 19%; the AUC of IPTT of DHB administrated decreased by about 15%, preferably decreased 21%, 25%, 27%, or 30%.

[0009] In some embodiments, compared with the untreated control group, the TG of DHB treated group was reduced by 10%, preferably reduced by 13%, 20%, or 26%; the TC of DHB treated group was reduced by 5%, preferably reduced by 9%, 15%, or 20%; the HDL of DHB treated group was increased by 5%, preferably increased by 8%, 15%, or 19%; the LDL of DHB treated group was reduced by 10%, preferably reduced by 15%, 18%, 25%, or 30%; especially the ratio of HDL / LDL of DHB treated group was increased by 20%, preferably increased by 30%, 50%, or 70%.

[0010] In some embodiments, a proportion of the dihydroberberine (DHB) is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%.

[0011] In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated in an amount of 1-2000 mg. In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated in an amount of 1-1500, 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.

[0012] In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated in an amount of 0.1 pM to 1 M or 0.01 to 5().0wt% (w / w). In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof may be administrated in an amount of 0.1 pM to 500 pM, 1 pM: to 500 pM, 1 pM to 5 mM, 1 pM to 500 mM, 5 pM to 500 pM, 5 pM to 5 mM, 5 pM to 100 mM, 5 pM to 500 mM, 50 pM to 500 pM, 50 pMto 5 mM. In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof may be administrated in an amount of 0.05% to 45%, 0.05%' io 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 9wt% (w / w).

[0013] In some embodiments, the subject is a mammal.

[0014] In some embodiments, the subject is human or animal. In some embodiments, the subject is human.

[0015] 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.

[0016] In some embodiments, the administration is through various routes selected from oral, intravenous injection, intramuscular injection, intraperitoneal injection, external use, or sublingual application.

[0017] 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.

[0018] In a second aspect, the present invention provides a composition comprising an effective amount of dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof, for improving or regulating GLP-1 level, or stimulating GLP-1 secretion in a subject.

[0019] 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.

[0020] In some embodiments, a proportion of the dihydroberberine (DHB) is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%', at least about 90%, at least about 95%, at least about 96%', at least about 97%, at least about 98%, or at least about 99%.

[0021] In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated at a daily dose of 1-2000 mg. In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt. polymer, ester, acid, or an analog or derivative thereof is administrated at a daily dose of 1-1500, 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.

[0022] In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated in an amount of 0.1 pM to 1 M or 0.01 to 50.0wt% (w / w). In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof may be administrated in an amount of 0.1 uM to 500 pM, 1 pM to 500 pM, 1 pM to 5 mM, 1 pM to 500 mM, 5 pM to 500 uM, 5 pM to 5 mM, 5 pM to 100 mM, 5 pM to 500 mM, 50 pM to 500 pM, 50 pM to 5 mM. In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof 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 9wt% (w / w).

[0023] In some embodiments, the subject is a mammal.

[0024] In some embodiments, the subject is human or animal. In some embodiments, the subject is human.

[0025] 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.

[0026] 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.

[0027] In a third aspect, the present invention provides use of a composition in a preparation of food, drink, supplement, biochemical composition, or nourishment for improving or regulating GLP-1 level, or stimulating GLP-1 secretion in a subject, wherein the composition comprises dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof.

[0028] 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.

[0029] In some embodiments, a proportion of the dihydroberberine (DHB) is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%', at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%.

[0030] In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated at a daily dose of 10-2000 mg. In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated at a daily dose of 1-1500, 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.

[0031] In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof is administrated in an amount of 0,1 pM to 1 M or 0.01 to 50.0wt% (w / w). In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof may be administrated in an amount of 0.1 pM to 500 pM, 1 pM to 500 pM, 1 pM to 5 mM, 1 pM to 500 mM, 5 pM to 500 pM, 5 pM to 5 mM, 5 pM to 100 mM, 5 pM to 500 mM, 50 pM to 500 pM, 50 pMto 5 mM. In some embodiments, the dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof 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 9wt% (w / w).

[0032] In some embodiments, the subject is a mammal.

[0033] In some embodiments, the subject is human or animal. In some embodiments, the subject is human.

[0034] In some embodiments, the administration is through various routes selected from oral, intravenous injection, intramuscular injection, intraperitoneal injection, external use, or sublingual application.

[0035] 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.

[0036] 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

[0037] Fig. 1 is percentage change curve of body weight of groups 1-4.

[0038] Fig. 2 is changes in the proportion of fat content in mice of groups 1 -4.

[0039] Fig. 3 is area under the curve (AUC) of oral glucose tolerance test (OGTT) in mice of groups 1-4.

[0040] Fig. 4 is area under the curve (AUC) insulin intraperitoneal tolerance test (IPTT) in mice of groups 1-4.

[0041] Fig. 5 is the fat pad weight of groups 1-4.

[0042] Fig. 6 is the insulin levels in serum of groups 1-4.

[0043] Fig. 7 is the TC levels in serum of groups 1-4.

[0044] Fig. 8 is the TG levels in serum of groups 1-4.

[0045] Fig. 9 is the LDL levels in serum of groups 1 -4.

[0046] Fig. 10 is the HDL levels in serum of groups 1-4.

[0047] Fig. 11 is the GLP-1 levels in serum of groups 1-4. DETAILED DESCRIPTION

[0048] 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.”

[0049] 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.

[0050] 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.

[0051] 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, and rate of excretion of dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative employed; 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.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] The dosage of dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof and / or composition comprising the same 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.

[0056] 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.

[0057] Dyslipidaemia or blood fat is associated with increased risk of cardiovascular disease. High Density Lipoprotein (HDL) is of clinical importance since an inverse correlation exists between plasma HDL concentrations and risk of atherosclerotic disease. The majority of cholesterol stored in atherosclerotic plagues originates from LDL and hence elevated concentrations Low' Density Lipoproteins (LDL) is closely associated with atherosclerosis. The HDL / LDL ratio is a clinical risk indictor for atherosclerosis and coronary atherosclerosis in particular. The invention can improve dyslipidemia by increasing the level of GLP-1 and then increasing the ratio of HDL / LDL.

[0058] 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.

[0059] As used herein, a “tonic” refers to a medicinal substance taken to give a feeling of vigor or well-being.

[0060] As used herein, a “syrup” refers to a thick sticky liquid derived from a sugar-rich plant, for example, sugar cane, corn, and maple.

[0061] 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.

[0062] “Intraperitoneal” as used here means within or administered through the peritoneum. The peritoneum is a thin, transparent membrane that li nes the walls of the abdominal (peritoneal) cavity and con tai ns / encl oses the abdominal organs such as the stomach and intestines.

[0063] As used herein, “sublingual” refers to situated or applied under the tongue.

[0064] A “functionalized food composition” includes a food composition that has a potentially positive effect on health beyond basic nutrition.

[0065] Various embodiments of the present invention provide for methods for administrating dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof to a subject for effectively improving or regulating GLP-1 level, 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 dihydroberberine may be administered to a human. DHB 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.

[0066] 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 dihydroberberine may be processed to increase bioavailability, solubility, palatability, and / or combination with other compounds.

[0067] Any titles or subheadings used herein are for organization purposes and should not be used to limit the scope of embodiments disclosed herein.

[0068] 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 ail 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.

[0069] 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. Examples

[0070] 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+DHB group (Group 3, n=8) and NC+DHB group (Group 4, n=8). The dosage of DHB supplementation is 1.5mg / kg. Ilie 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 group 3 also continued to eat a high-fat diet and received daily intragastric administration of sample for 8 weeks; the group 4 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°C for storage.

[0071] Measurement of Blood Glucose: Oral glucose tolerance test (OGTT): At week 16th of the experiment, after fasting for 15h, 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 15h, insulin (lU / kg) was injected intraperitoneally and blood glucose was measured at 0, 15, 30, 60, 90, 120 mins.

[0072] Body composition analysis: During the experiment, the fat mass and lean mass was measured by Nuclear Magnetic Resonance Body Composition Analyzer once a week.

[0073] 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°C 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.

[0074] 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.

[0075] Change of body weight: Weigh once a week at a fixed time from beginning of the experiment.

[0076] Figure 1 is percentage change curve of body weight of groups 1-4. In figure 1, after taking DHB for 8 weeks, body weight in HFD+DHB group (33.36 g) decreased by 9.84%' compared with HFD group (37 g), NC+DHB group (31.33 g) decreased by 7.75%' at the 8th week compared with NC group (33.96 g). Thus, the body weight curve showed that DHB groups were helpful in inhibiting high fat induced obesity. Compared with the untreated control group, the body weight of DHB treated group was decreased approximately 5%, preferably decreased 7%, 9%. Figure 2 is changes in the proportion of fat content in mice of groups 1-4. The high body fat content is highly suggestive of possible obesity or abnormal endocrine system. As shown in figure 2, mice in group 3 and 4 were more effective in inhibiting obesity, the fat content in HFD+DHB group (Group 3) was decreased by approximately 19% as compared to HFD group (Group 2), and the fat content in NC+DHB group (Group 4) was decreased approximately by 35% as compared to control mice (Group 1). Figure 3 is area under the curve (AUC) of oral glucose tolerance test (OGTT) in mice of groups 1-4. Figure 4 is area under the curve (AUC) insulin intraperitoneal tolerance test (IPTT) in mice of groups 1-4.

[0077] In figure 3, the AUC of OGTT in HFD+DHB group (Group 3) was decreased by approximately 19% as compared to HFD group (Group 2), and the AUC of OGTT in NC+DHB group (Group 4) was decreased by approximately 10% as compared to control mice (Group 1). From figure 4, the AUC of IPTT in HFD+DHB group (Group 3) was decreased by about 27% as compared to HFD group (Group 2), and the AUC of IPTT in NC+DHB group (Group 4) was decreased by approximately 21% as compared to control mice (Group 1). DHB supplementation could improve the blood glucose and insulin tolerance relative to mice without receiving DHB supplementation.

[0078] From figures 1 -4, the results indicated that supplementing DHB can achieve the beneficial effects on metabolic health than without DHB supplementation. In particular, compared with the untreated control group (HFD group), the body weight of DHB treated group was decreased approximately 5%, preferably decreased 7%, 9%; the fat content of DHB treated group reduced by approximately 15%, preferably decreased 19%, 30%, 35%; and the AUC of OGTT of DHB supplemented decreased by about 5%, preferably decreased 10%, 15%, 19%; the AUC of IPTT of DHB administrated decreased by about 15%, preferably decreased 21%, 25%, 27%, or 30%.

[0079] Figure 5 is the fat pad weight of groups 1-4. In figure 5, DHB supplementation could significantly increase the pararenal fat pad weight relative to mice without receiving DHB supplementation. Figures 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. 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.

[0080] From the point of view of serum indexes (Figures 6-11), TG results showed a 262 + reduction in serum TG of HFD+DHB group compared with HFD group and a 13.2% reduction in NC+DHB group compared with NC group. From the results of TC, blood TC in HFDrDHB group was reduced by 20.6% compared with HFD group and 9.8% in NC+DHB group compared with NC group. HDL results showed a 19% increase in serum HDL of HFD+DHB group compared with HFD group and an 8.8% increase in NC+DHB group compared with NC group. As for the LDL, compared with the HFD group, HFD+DHB group showed a 32.3% decrease, and the NC+DHB group showed a 18.3% decrease compared with NC group. In addition, as for the ratio of HDL / LDL, compared with the HFD group, HFD+DHB group showed a 76% increase, and the NC+DHB group showed a 33% increase compared with NC group. Therefore, DHB has good effects on INS, GLP-1, LDL and HDL. Compared with the untreated control group, the TG of DHB treated group was reduced by 10%, preferably reduced by 13%, 20%, 26%; the TC of DHB treated group was reduced by 5%, preferably reduced by 9%, 15%, 20%; the HDL of DHB treated group was increased by 5%, preferably increased by 8%, 15%, 19%; the LDL of DHB treated group was reduced by 10%, preferably reduced by 15%, 18%, 25%, 30%; especially the ratio of HDL / LDL of DHB treated group was increased by 20%, preferably increased by 30%, 50%, 70%; the GLP-1 of DHB treated group was increased by 10%, preferably increased by 15%, 20%, 30%, 50%, 70%, 80%, 90%, 100%. DHB supplementation could significantly increase HDL and GLP-1, and decrease INS, TC, TG, and LDL relative to mice without receiving DHB supplementation, indicating that DHB has the effect of improving obesity and ameliorating metabolism. This effect persists for as long as the test protocol is continued.

[0081] 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 stimulating GLP-1 secretion in a subject, comprising administrating to the subject a composition comprising: an effective amount of dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof.

2. The method of claim 1, wherein the method is used for ameliorating metabolism including losing weight, managing glucose or insulin tolerance, or for ameliorating blood fat, 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, compared with the untreated control group, the GLP-1 of DHB treated group was at least increased by 10%.

4. The method of claim 1 or 2, wherein the said ameliorating blood fat is the increased ratio of HDL / LDL.

5. The method of any of claims 1-4, wherein a proportion of the dihydroberberine (DHB) is at leastabout 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, or at least about 99%.

6. The method of any of claims 1-5, wherein the dihydroberberine, or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative is administrated in an amount of 1 -2000 mg.

7. The method of any of claims 1-6, wherein the Dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative is administrated in an amount of 0.1 pMto 1 M or 0.01 to 50.0wt% (w / w).

8. The method of any of claims 1-7, wherein the subject is a mammal.

9. The method of any of claims 1-8, wherein the subject is human or animal.

10. The method of any of claims 1-9, wherein the composition is prepared as a food, a drink, a supplement, a biochemical composition, animal’s food, or a nutraceutical composition.

11. The method of any of claims 1-10, wherein the administration is through various routes selected from oral, intravenous injection, intramuscular injection, intraperitoneal injection, external use, or sublingual application.

12. The method of any of claims 1-11, 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.

13. A composition comprising an effective amount of dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof, for improving or regulating GLP-1 level, or stimulating GLP-1 secretion in a subject.14 The composition of claim 13, 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.

15. The composition of claim 13 or 14, wherein a proportion of the dihydroberberine (DHB) is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%.

16. The composition of any of claims 13-15, wherein the dihydroberberine, or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative is administrated at a daily dose of 1-2000 mg.

17. Use of a composition in a preparation of food, drink, supplement, biochemical composition, or nourishment for improving or regulating GLP-1 level, or stimulating GLP-1 secretion in a subject, wherein the composition comprises dihydroberberine (DHB), or a physiologically acceptable salt, polymer, ester, acid, or an analog or derivative thereof.

18. The use of claim 17, 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.

19. The use of claim 17 or 18, wherein the subject is a mammal.

20. The use of any of claims 17-19, wherein a proportion of the dihydroberberine (DHB) is at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, or at least about 99%.

21. The use of any of claims 17-20, wherein the dehydroascorbic acid or a stereoisomer, tautomer, hydrate, salt, ester, or solvate thereof is administrated at a daily dose of 1-2000 mg or in an amountof 0.1 pM to 1 M or 0.01 to 50.0wt% (w / w).

22. The use of any of claims 17-21, 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.

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