Use of poria cocos periderm extract, poria acid A, and poria acid B for regulating blood glucose

Abstract

A use of at least one of poricoic acid A and poricoic acid B in the preparation of a medicament or a health food, wherein the medicament or the health food is used for regulating blood sugar. At least one of poricoic acid A and poricoic acid B can be used in the form of a plant extract, wherein the total content of poricoic acid A and poricoic acid B in the plant extract is not less than 30% by weight based on the total weight of the plant extract.

Description

[0001] This application is a divisional application. The original application was filed on May 9, 2017, with application number 201710320602.9, and the invention title was "Use of Poria cocos peel extract, pachymanic acid A, and pachymanic acid B in regulating blood sugar". Technical Field

[0002] This invention relates to the application of Poria cocos peel extract, poricoic acid A, and poricoic acid B, particularly to the application of these substances in regulating blood sugar. Background Technology

[0003] Diabetes is a chronic metabolic disorder primarily caused by abnormalities in the body's cellular glucose uptake mechanisms, leading to excessively high blood glucose levels. Normally, insulin secreted by the beta cells of the pancreas stimulates fat and muscle cells to absorb glucose, regulating blood sugar levels. When an organism experiences insufficient insulin secretion or poor insulin sensitivity due to factors such as obesity or aging, blood sugar levels rise. High blood sugar can lead to complications such as high blood pressure, heart disease, arteriosclerosis, and high cholesterol; in severe cases, it can cause long-term consequences such as blindness, impotence, amputation, and the need for dialysis.

[0004] Currently, clinical treatments for diabetes primarily include exercise, dietary control, and medication. Medication includes insulin injections and oral hypoglycemic agents, such as sulfonylureas, biguanides, alpha-glucosidase inhibitors, and insulin sensitizers. However, with changing lifestyles, the prevalence of diabetes is increasing annually. According to a 2008 World Health Organization forecast, the global diabetic population is projected to exceed 300 million by 2030. Therefore, the industry continues to focus on developing drugs or methods with fewer side effects and effective blood sugar control.

[0005] The inventors of this application have discovered that the extract of Poria cocos peel and its contained poricoic acid A and poricoic acid B can effectively enhance the cells' ability to take up glucose, and therefore can be used to regulate blood sugar, especially providing excellent blood sugar lowering effects. Summary of the Invention

[0006] One object of the present invention is to provide the use of at least one of pachyrhizonic acid A and pachyrhizonic acid B in the manufacture of a drug or health food, wherein the drug or health food is used to regulate blood sugar. The dosage of the drug or health food, based on the total weight of pachyrhizonic acid A and pachyrhizonic acid B, is approximately 0.05 mg / kg body weight to 1 mg / kg body weight per day.

[0007] Preferably, at least one of pachymic acid A and pachymic acid B is used in the form of a plant extract. In this plant extract, the total content of pachymic acid A and pachymic acid B, based on the total weight of the plant extract, is not less than 30% by weight, preferably not less than 40% by weight. More preferably, at least one of pachymic acid A and pachymic acid B is used in the form of a Poria cocos peel extract. In this Poria cocos peel extract, based on the total weight of the Poria cocos peel extract, the content of each of pachymic acid, dehydropachymic acid, tumulosic acid, and dehydrotumulosic acid does not exceed 0.5% by weight, and the content of each of dehydrotrametenolic acid, trametenolic acid, dehydroeburicoic acid, and eburicoic acid does not exceed 5%. Preferably, in the Poria cocos peel extract, the content of each of dehydropyrrolizic acid, pyrrolizic acid, dehydroporinic acid, and porinic acid, based on the total weight of the Poria cocos peel extract, does not exceed 2.5%. More preferably, in the Poria cocos peel extract, the content of each of dehydropyrrolizic acid, pyrrolizic acid, dehydroporinic acid, and porinic acid, based on the total weight of the Poria cocos peel extract, does not exceed 1%.

[0008] Another object of the present invention is to provide a method for regulating blood glucose in an individual, comprising administering to an individual in need an effective amount of at least one of pachymetic acid A and pachymetic acid B.

[0009] Another object of the present invention is to provide a blood sugar regulating composition, which is a medicine or health food and contains an effective amount of at least one of pachymetic acid A and pachymetic acid B.

[0010] Preferably, the Poria cocos peel extract is provided by an operation comprising the following steps:

[0011] (a) Extracting a piece of Poria cocos peel with a first solvent to obtain a crude extract;

[0012] (b) Dry the crude extract to obtain a crude extract powder;

[0013] (c) Extract the crude extract powder with a second solvent to obtain a Poria cocos peel extract.

[0014] The first solvent may be the same as or different from the second solvent and is selected from water, ethanol, alkaline solution, acid solution, and combinations thereof.

[0015] The first solvent and the second solvent are aqueous solutions of ethanol with the same or different concentrations.

[0016] The beneficial effects of the present invention are: the extract of Poria cocos peel, as well as Poria cocos acid A and Poria cocos acid B, can indeed enhance the ability of biological cells to take up blood sugar, and can be used to regulate blood sugar, especially to lower excessively high blood sugar. Attached Figure Description

[0017] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0018] Figure 1 The results show the effect of Poria cocos peel extract on the ability of mouse muscle cells to take up glucose from culture medium, as detected by the glucose oxidase assay.

[0019] Figure 2 The results show the effect of Poria cocos peel extract on the ability of mouse adipocytes to take up glucose from culture medium, as determined by the glucose oxidase method.

[0020] Figure 3A The results show the effect of Poria cocos acid A on the ability of mouse muscle cells to take up glucose from the culture medium, as determined by the glucose oxidase method.

[0021] Figure 3B The results show the effect of Poria cocos acid B on the ability of mouse muscle cells to take up glucose from the culture medium, as determined by the glucose oxidase method.

[0022] Figure 4A The results show the effect of Poria cocos acid A on the ability of mouse adipocytes to take up glucose from the culture medium, as determined by the glucose oxidase method.

[0023] Figure 4B The results show the effect of Poria cocos acid B on the ability of mouse adipocytes to take up glucose from the culture medium, as determined by the glucose oxidase method. Detailed Implementation

[0024] The following describes some specific embodiments of the present invention; however, the present invention can be practiced in many different forms without departing from its spirit, and the scope of protection of the present invention should not be construed as limited to the contents stated in the specification. Furthermore, unless otherwise stated herein, the terms “a,” “the,” and similar terms used in this specification (especially in the claims) should be understood to include both singular and plural forms; “effective amount” refers to the amount of compound that, when administered to an individual, can effectively enhance the ability of that individual’s cells to take up glucose; “individual” refers to a mammal, which can be human or non-human animal; “regulating blood glucose” refers to changing the blood glucose concentration toward normal values; the unit “mg / kg body weight” refers to the dosage required per kilogram of body weight per individual.

[0025] The numerical ranges used in this specification (e.g., 5 to 100) should be understood to include all rational numbers within that range and any range of rational numbers within that range. Therefore, the numerical ranges used in this specification include all possible combinations of values ​​between the listed minimum and maximum values. Furthermore, when the word "about" is used before a numerical value, it substantially means that the value differs from the stated value by no more than 20%, preferably no more than 10%, and more preferably no more than 5%.

[0026] As explained above, the main cause of diabetes is an abnormality in the mechanism by which cells in the body take up glucose, leading to excessively high blood glucose levels. The inventors of this application have discovered that both pachymetic acid A and pachymetic acid B can enhance the ability of cells to take up glucose, and therefore can be used to regulate blood sugar, especially to lower excessively high blood sugar. Therefore, this invention provides an application of at least one of pachymetic acid A and pachymetic acid B in regulating blood sugar, including using at least one of pachymetic acid A and pachymetic acid B in the preparation of a blood sugar regulating drug or health food, administering at least one of pachymetic acid A and pachymetic acid B to individuals in need to regulate blood sugar, and providing a health food or pharmaceutical composition containing at least one of pachymetic acid A and pachymetic acid B.

[0027] Poria cocos (Schw.) Wolf is a medicinal herb containing the dried sclerotium of a fungus belonging to the family Poriaceae. This fungus commonly parasitizes pine roots, with a light brown or dark brown outer skin (the pericarp) and a pink or white interior (the flesh). Traditional Chinese medicine texts record that the flesh of Poria cocos has sedative, diuretic, nutritional, immune-boosting, and anti-aging properties, while the pericarp is only used to treat skin edema.

[0028] As shown in the accompanying embodiments, according to the present invention, a Poria cocos peel extract containing a total content of not less than 30% by weight (based on the total weight of the Poria cocos peel extract) of pachymica A and pachymica B can be obtained from the Poria cocos peel. Therefore, at least one of pachymica A and pachymica B used according to the present invention can be used, for example, in the form of a plant extract of Poria cocos peel, wherein, in the plant extract used according to the present invention, the total content of pachymica A and pachymica B, based on the total weight of the plant extract, is not less than 30% by weight, preferably not less than 40% by weight.

[0029] According to the present invention, the Poria cocos peel extract used can be an extract provided by an operation comprising the following steps: (a) extracting the Poria cocos peel with a first polar solvent to obtain a crude extract; (b) drying the crude extract to obtain a crude extract powder; and (c) extracting the crude extract powder with a second polar solvent to obtain a Poria cocos peel extract, wherein the first polar solvent and the second polar solvent are the same or different and are respectively selected from water, ethanol, alkaline solution, acid solution, and combinations thereof. Alkaline solution refers to any suitable alkaline solution with a pH greater than 7 (e.g., sodium hydroxide solution), and acid solution refers to any suitable acidic solution with a pH less than 7 (e.g., hydrochloric acid solution). In some embodiments of the present invention, aqueous solutions of ethanol with the same or different concentrations are used as the first polar solvent and the second polar solvent.

[0030] In step (a), the ratio of the first polar solvent to the Poria cocos peel can be adjusted as needed. Generally, there is no particular limitation on the amount of the first polar solvent, as long as it can ensure uniform dispersion of the raw materials. For example, in step (a), the volume ratio of the first polar solvent to the Poria cocos peel can be approximately 8:1 to approximately 16:1. In a specific embodiment of the present invention, an aqueous ethanol solution is used as the first polar solvent, and the extraction in step (a) is performed with a volume ratio of Poria cocos peel to aqueous ethanol solution of approximately 1:8.

[0031] In step (a), the appropriate extraction time can be selected based on the first polar solvent used. For example, using an aqueous ethanol solution as the first polar solvent with a volume ratio of Poria cocos peel to aqueous ethanol solution of 1:8, the extraction time is typically at least 1 hour, preferably at least 2 hours, and more preferably at least 3 hours. Furthermore, other operations such as decoction, cooling, filtration, vacuum concentration, and resin column chromatography can be performed as needed to carry out step (a). Additionally, if necessary, the Poria cocos peel can be pre-soaked in the first polar solvent for a period of time before performing step (a). For example, using an aqueous ethanol solution as the first polar solvent, pre-soaking for approximately 12 hours is recommended.

[0032] In step (c), the ratio of the second polar solvent to the crude extract powder obtained in step (b) can be adjusted as needed. Generally, there are no particular limitations on the amount of the second polar solvent, as long as it can uniformly disperse the crude extract powder. For example, in step (c), the volume ratio of the second polar solvent to the crude extract powder of Poria cocos peel can be from about 8:1 to about 16:1. In a specific embodiment of the present invention, an aqueous ethanol solution is used as the second polar solvent, and the extraction in step (c) is performed with a volume ratio of crude extract powder of Poria cocos peel to aqueous ethanol solution of about 1:8.

[0033] The Poria cocos peel extract used according to the present invention can also be a dried product, which can be provided by drying the extract obtained in step (c). To achieve the greatest possible extraction efficiency, if necessary, the Poria cocos peel can be repeatedly extracted with the same or different first polar solvents before step (b), and the extracts obtained from the multiple extractions can be combined to provide the crude extract for step (b); steps (b), (c), and other operations described above as needed can also be repeated in cycles.

[0034] In the Poria cocos peel extract used according to the present invention, the total content of pamoate A and pamoate B, based on the total weight of the Poria cocos peel extract, is not less than 30% by weight, preferably not less than 40% by weight, and the content of each of pamoateic acid, dehydropamoateic acid, tomolatesic acid, and dehydrotomolatesic acid does not exceed 0.5%, and the content of each of dehydropyrrolizic acid, pyrrolizic acid, dehydroporphyrinsic acid, and porphyrinsic acid does not exceed 5%. Preferably, in the Poria cocos peel extract, based on the total weight of the Poria cocos peel extract, the content of each of dehydropyrrolizic acid, pyrrolizic acid, dehydroporphyrinsic acid, and porphyrinsic acid does not exceed 2.5%. More preferably, in the Poria cocos peel extract, based on the total weight of the Poria cocos peel extract, the content of each of dehydropyrrolizic acid, pyrrolizic acid, dehydroporphyrinsic acid, and porphyrinsic acid does not exceed 1%.

[0035] The pharmaceutical composition or drug provided according to the invention may be in any suitable form without particular limitation, depending on the intended use and the appropriate dosage form. For example, but not limited thereto, the pharmaceutical composition or drug may be administered to the individual in need by oral or non-oral route (e.g., subcutaneous, intravenous, intramuscular, or intraperitoneal). Depending on the form of use and purpose, a suitable carrier may be selected to provide the pharmaceutical composition or drug, including excipients, diluents, adjuvants, stabilizers, absorption delayers, disintegrants, solubilizers, emulsifiers, antioxidants, binders, binders, thickeners, dispersants, suspending agents, lubricants, hygroscopic agents, etc.

[0036] Taking a dosage form suitable for oral administration as an example, the pharmaceutical composition or drug provided according to the invention may contain any pharmaceutically acceptable carrier that will not adversely affect the desired efficacy of the active ingredient (i.e., pachymetic acid A, pachymetic acid B, or poria cocos peel extract), such as: water, saline, glucose, dextrose, glycerol, ethanol or analogues thereof, cellulose, starch, sugar bentonite, and combinations thereof. The pharmaceutical composition or drug may be provided in a dosage form suitable for oral administration using any suitable method, such as: tablets (e.g., sugar-coated tablets), pills, capsules, granules, powders, fluid extracts, solutions, syrups, suspensions, tinctures, etc.

[0037] As for injections or infusions suitable for subcutaneous, intravenous, intramuscular, or intraperitoneal administration, the pharmaceutical composition or drug provided according to the invention may contain one or more components such as isotropic solutions, salt buffers (e.g., phosphate buffer or citrate buffer), solubilizers, emulsifiers, 5% sugar solutions, and other carriers, and may be provided in dosage forms such as intravenous infusions, emulsion intravenous infusions, dry powder injections, suspension injections, or dry powder suspension injections. Alternatively, the pharmaceutical composition or drug may be prepared as a pre-injection solid, provided in a dosage form soluble in other solutions or suspensions, or in an emulsifiable dosage form, and the pre-injection solid may be dissolved in other solutions or suspensions or emulsified before being administered to the individual in need, to provide the desired injection.

[0038] As needed, the pharmaceutical composition or drug provided according to the present invention may further contain appropriate amounts of additives, such as flavoring agents, coloring agents, colorants, etc., to improve the taste and visual appeal of the pharmaceutical composition or drug during administration, and buffers, preservatives, antiseptics, antibacterial agents, antifungal agents, etc., to improve the stability and storability of the drug. Furthermore, the pharmaceutical composition or drug may, as needed, contain one or more other active ingredients (e.g., antioxidants, insulin sensitizers, etc.), or be used in combination with drugs containing one or more other active ingredients, to further enhance the efficacy of the pharmaceutical composition or drug or increase the flexibility and compatibility of the formulation, provided that the other active ingredients do not adversely affect the efficacy of the active ingredient of the present invention (i.e., pachymica A, pachymica B, or poria cocos peel extract).

[0039] The pharmaceutical composition or drug provided according to the present invention can be administered at different frequencies, such as once a day, multiple times a day, or once every few days, depending on the individual's needs, age, weight, and health condition. For example, when administered orally to an individual to regulate blood sugar, the dosage, based on the total weight of pachymansin A and pachymansin B, is approximately 0.01 mg / kg body weight to 5 mg / kg body weight per day, preferably approximately 0.03 mg / kg body weight to 2 mg / kg body weight per day, and more preferably approximately 0.05 mg / kg body weight to 1 mg / kg body weight per day. Alternatively, based on the extract of Poria cocos peel, the dosage is approximately 0.025 mg / kg body weight to 25 mg / kg body weight per day, preferably approximately 0.075 mg / kg body weight to 10 mg / kg body weight per day, and more preferably approximately 0.125 mg / kg body weight to 5 mg / kg body weight per day.

[0040] The health food products provided by the present invention can be made into products such as dairy products, processed meat products, bread, pasta products, biscuits, lozenges, capsules, fruit juices, teas, sports drinks, and nutritional drinks, but are not limited thereto.

[0041] The health food provided according to the present invention can be consumed at different frequencies, such as once a day, multiple times a day, or once every few days, depending on the recommended intake based on the individual's age, weight, and health condition. The content of pachymetic acid A, pachymetic acid B, or poria cocos peel extract in the health food provided by the present invention can also be adjusted for specific groups, preferably to the recommended daily intake. For example, based on the total weight of pachymetic acid A and pachymetic acid B, if an individual's recommended intake is approximately 70 mg of pachymetic acid A and pachymetic acid B per day, and each serving of the health food contains a total of 35 mg of pachymetic acid A and pachymetic acid B, then that individual can consume approximately two servings of the health food per day.

[0042] The outer packaging of the health food of this invention can indicate the recommended dosage, the usage standards and conditions for specific groups (such as pregnant women, diabetic patients, and patients with kidney disease), or the recommendations for taking it with other foods or medicines, so that users can take it at home without any safety concerns without the guidance of a doctor, pharmacist or relevant staff.

[0043] The present invention also provides a method for regulating blood glucose, comprising administering an effective amount of an active ingredient to an individual in need, wherein the active ingredient is at least one of pachymetic acid A and pachymetic acid B. In the method for regulating blood glucose according to the present invention, the state, route of administration, form of administration, applicable dosage, and related therapeutic application of the active ingredient are as described above.

[0044] The present invention will now be further illustrated by the following embodiments. These embodiments are provided for illustrative purposes only and are not intended to limit the scope of protection of the invention. The scope of protection of the invention is as described in the claims.

[0045] Example

[0046] [Preparation Examples]

[0047] A. Preparation of Poria cocos peel extract

[0048] A-1. Take Poria cocos (originating from Yunnan), wash it, and peel off its outer skin (hereinafter referred to as "Poria cocos peel"), leaving the flesh (hereinafter referred to as "Poria cocos flesh"). Take the aforementioned Poria cocos peel and soak it in a 75% ethanol aqueous solution at room temperature at a volume ratio of 1:8 (Poria cocos: ethanol aqueous solution) for 12 hours, then boil and extract (for 3 hours). Repeat the above extraction steps three times. Combine the extracts obtained from the three extractions and filter to remove insoluble matter to obtain a crude extract. Next, concentrate the crude extract under reduced pressure to remove the solvent, and then dry it using a spray dryer to obtain a crude extract powder.

[0049] A-2. Take the crude extract powder obtained in A-1, mix it with 95% ethanol at a volume ratio of 1:8 (crude extract powder: ethanol aqueous solution), and extract it (for 3 hours). Then, separate it using a column with silica gel as the stationary phase to obtain a Poria cocos peel extract.

[0050] A-3. Take the crude extract powder obtained in A-1 and disperse it uniformly in pure water at a volume ratio of 1:10 (crude extract powder: water). Next, add sodium hydroxide to raise the pH of the mixture to about 12, then pour it into a mixing tank maintained at 65°C and stir evenly until the reaction is complete. Then, neutralize with 12N concentrated hydrochloric acid, centrifuge to remove the filtrate, wash the remaining insoluble matter with pure water, and then dry the insoluble matter with a spray dryer to obtain an extract powder. Take the aforementioned extract powder and extract it three times with 1N sodium bicarbonate aqueous solution at a volume ratio of 1:20 (extract powder: 1N sodium bicarbonate aqueous solution). Combine the extracts obtained from the three extractions, neutralize with 12N concentrated hydrochloric acid, centrifuge to remove the filtrate, wash the remaining insoluble matter with pure water, and then dry the insoluble matter with a spray dryer to obtain a Poria cocos peel extract.

[0051] A-4. The components of the extracts obtained in A-2 and A-3 were detected by liquid chromatography / ultraviolet light / mass spectrometry at wavelengths of 243 nm and 210 nm, respectively. The content of each component in each extract was quantified by high performance liquid chromatography. The analytical results of the extract obtained in A-2 are shown in Table 1, and the analytical results of the extract obtained in A-3 are shown in Table 2.

[0052] Table 1

[0053]

[0054]

[0055] Table 2

[0056] Pachymic acid (PA) － Dehydropachymic acid (DPA) － tumulosic acid (TA) － Dehydrotumulosic acid (DTA) 0.31 Polyporenic acid C (PAC) 1.15 3-Epi-dehydrotumulosic acid (EDTA) 0.40 Dehydrotrametenolic acid (DTTA) 0.58 Trametenolic acid (TTA) 0.35 Poria cocoic acid A (PAA) 41.06 Dehydroeburicoic acid (DEA) 0.19 Poria cocoic acid B (PAB) 16.50 eburicoic acid (EA) 0.20

[0057] As shown in Table 1, the Poria cocos peel extract obtained by A-2 contains high amounts of pachymonic acid A (32.72% by weight of the extract) and pachymonic acid B (10.44% by weight of the extract), low amounts of dehydrosporinic acid, sporinic acid, dehydroporinic acid and porinic acid (2.01%, 0.85%, 1.2%, and 0.83% by weight of the extract, respectively), and very low amounts of dehydrotomolic acid (0.46% by weight of the extract), but does not contain pachymic acid, dehydropachymic acid and tomolic acid.

[0058] As shown in Table 2, the Poria cocos peel extract obtained from A-3 contains high amounts of pachymonic acid A (41.06% by weight of the extract) and pachymonic acid B (16.50% by weight of the extract), low amounts of pachymic acid C and dehydropachymic acid (1.15% and 0.58% by weight of the extract, respectively), and very low amounts of dehydrotomolic acid, 3-epi-dehydrotomolic acid, pachymic acid, dehydroporphyric acid, and porphyric acid (0.31%, 0.40%, 0.35%, 0.19%, and 0.20% by weight of the extract, respectively), but does not contain pachymic acid, dehydropachymic acid, or pachymic acid.

[0059] B. Preparation of Poria cocos acid A and Poria cocos acid B

[0060] B-1. Dissolve the Poria cocos peel extract obtained in A-2 or A-3 uniformly in methanol at a volume ratio of 1:500 (Poria cocos peel extract: methanol). After filtration to remove insoluble matter, separate and collect pachymetic acid A and pachymetic acid B using a preparative high-performance liquid chromatography (using a methanol-water mixture as the mobile phase) at a wavelength of 243 nanometers. After collection, remove methanol using a vacuum concentrator to obtain pachymetic acid A and pachymetic acid B, respectively.

[0061] B-2. The purity of porphyrin A and porphyrin B obtained in B-1 was detected by liquid chromatography / ultraviolet light / mass spectrometry at a wavelength of 243 nanometers. The results showed that the purity of porphyrin A and porphyrin B was greater than 98%.

[0062] C. Cell Culture

[0063] Differentiated mouse muscle cells (C2C12, purchased from ATCC) and adipocytes (3T3-L1, purchased from ATCC) were cultured separately in Dulbecco's modified Eagle's medium (DMEM) containing 2% bovine serum albumin (BSA) but without serum for 16 hours. They were then washed with Dulbecco's phosphate-buffered saline (D-PBS), and the medium was replaced with 0.2% BSA-DMEM containing 500 μg / mL glucose for subsequent experiments.

[0064] Example 1: Effect of Poria cocos peel extract on cellular glucose uptake capacity

[0065] (1-1) Muscle cells

[0066] The C2C12 cells provided in the [Preparation Example] were divided into five groups and cultured in the following culture media for 2 hours:

[0067] Group I: BSA-DMEM medium containing 500 μg glucose / mL;

[0068] 2. Group II: BSA-DMEM medium containing 500 μg glucose / mL and insulin at a concentration of 100 nanomoles;

[0069] 3. Group III: BSA-DMEM medium containing 500 μg glucose / mL and a solution of Poria cocos peel extract provided in [Preparation Example A-2] at a concentration of 0.1 μg / mL;

[0070] 4. Group IV: BSA-DMEM medium containing 500 μg glucose / mL and a solution of Poria cocos peel extract provided in [Preparation Example A-2] at a concentration of 1 μg / mL;

[0071] 5. Group V: BSA-DMEM medium containing 500 μg glucose / mL and a solution of Poria cocos peel extract provided in [Preparation Example A-2] at a concentration of 10 μg / mL.

[0072] Subsequently, the glucose content in the cell culture medium of each group was determined using the glucose oxidase method to understand the degree of glucose consumption in each group (representing the cells' ability to take up glucose). Finally, based on the results of the control group (i.e., cells cultured in group I culture medium), the relative glucose uptake capacity of the other groups was calculated, and the results are shown below. Figure 1 .

[0073] Depend on Figure 1 It can be seen that, compared with the control group, the glucose uptake capacity of cells treated with the Poria cocos peel extract of the present invention (i.e., cells cultured in group III, IV, or V culture medium) was significantly improved, even surpassing that of the positive control group (i.e., cells cultured in group II culture medium). The foregoing results show that the Poria cocos peel extract of the present invention can effectively enhance the glucose uptake capacity of muscle cells, and therefore can be used to regulate blood sugar.

[0074] (1-2) Adipocytes

[0075] 3T3-L1 cells provided in the [Preparation Example] were divided into five groups and cultured in the culture media of groups I to V as described in Example 1 (1-1) for 2 hours. Next, the glucose content in the cell culture medium of each group was measured using the glucose oxidase method to understand the degree of glucose consumption (representing the cells' ability to take up glucose). Finally, based on the results of the control group (i.e., cells cultured in group I culture medium), the relative glucose uptake capacity of the other groups was calculated, and the results are shown below. Figure 2 .

[0076] Depend on Figure 2 It can be seen that, compared with the control group, the glucose uptake capacity of cells treated with the Poria cocos peel extract of the present invention (i.e., cells cultured in group III, IV, or V culture medium) all showed an increasing trend, with the most significant increase observed in the high-dose group (cells cultured in group V culture medium). The aforementioned results indicate that the Poria cocos peel extract of the present invention can effectively enhance the glucose uptake capacity of adipocytes, and therefore can be used to regulate blood sugar.

[0077] Example 2: Effects of Poria cocos acid A and Poria cocos acid B on cellular glucose uptake capacity

[0078] (2-1) Muscle cells

[0079] The C2C12 cells provided in the [Preparation Example] were divided into eight groups and cultured in the following culture media for 2 hours:

[0080] 1. Group i: BSA-DMEM medium containing 500 μg glucose / mL;

[0081] 2. Group ii: BSA-DMEM medium containing 500 μg glucose / mL and insulin at a concentration of 100 nanomoles;

[0082] 3. Group iii: BSA-DMEM medium containing 500 μg glucose / mL and 0.1 μg / mL of pachymic acid A or pachymic acid B provided in [Preparation Example B-1];

[0083] 4. Group IV: BSA-DMEM medium containing 500 μg glucose / mL and 1 μg / mL of pachymic acid A (or pachymic acid B) provided in [Preparation Example B-1];

[0084] 5. Group v: BSA-DMEM medium containing 500 μg / mL glucose and 10 μg / mL of pachymic acid A (or pachymic acid B) provided in [Preparation Example B-1].

[0085] Next, the glucose content in the cell culture medium of each group was determined using the glucose oxidase method to understand the glucose consumption level of each group (representing the cells' ability to take up glucose). Finally, based on the results of the control group (i.e., cells cultured in the i-th group's culture medium), the relative glucose uptake capacity of the other groups was calculated. The results are shown below. Figure 3A and Figure 3B .in, Figure 3A Results include the control group, the positive control group (i.e., cells cultured in group ii culture medium), and the group treated with pachymic acid A (i.e., cells cultured in group iii, iv, or v culture medium containing pachymic acid A). Figure 3B The results include the control group, the positive control group, and the group treated with pachymectin B (i.e., cells cultured in the culture medium of group iii, iv, or v, wherein the culture medium contains pachymectin B).

[0086] Depend on Figure 3A It can be seen that, compared with the control group, the glucose uptake capacity of the group treated with Poria cocos acid A was significantly improved to the same level as the control group. The above results show that Poria cocos acid A can effectively enhance the ability of muscle cells to uptake glucose, and therefore can be used to regulate blood sugar.

[0087] Depend on Figure 3B It can be seen that, compared with the control group, the glucose uptake capacity of the group treated with Poria cocos B was significantly improved. The aforementioned results show that Poria cocos B can also regulate blood sugar by enhancing the ability of muscle cells to uptake glucose.

[0088] (2-2) Adipocytes

[0089] 3T3-L1 cells provided in the [Preparation Example] were divided into eight groups and cultured in the culture media of groups i to v as described in Example 2 (2-1) for 2 hours. Next, the glucose content in the cell culture medium of each group was measured using the glucose oxidase method to understand the degree of glucose consumption (representing the cells' ability to take up glucose). Finally, based on the results of the control group (i.e., cells cultured in group i), the relative glucose uptake capacity of the other groups was calculated, and the results are shown below. Figure 4A and Figure 4B.in, Figure 4A Results include the control group, the positive control group (i.e., cells cultured in group ii culture medium), and the group treated with pachymic acid A (i.e., cells cultured in group iii, iv, or v culture medium containing pachymic acid A). Figure 4B The results include the control group, the positive control group, and the group treated with pachymectin B (i.e., cells cultured in the culture medium of group iii, iv, or v, wherein the culture medium contains pachymectin B).

[0090] Depend on Figure 4A It was found that, compared with the control group, the glucose uptake capacity of the groups treated with Poria cocos A showed an increasing trend, with the increase being most significant in the medium-dose group (i.e., cells cultured in group iv) and the high-dose group (i.e., cells cultured in group v). These results indicate that Poria cocos A can effectively enhance the glucose uptake capacity of adipocytes, and therefore can be used to regulate blood sugar.

[0091] Depend on Figure 4B It can be seen that, compared with the control group, the glucose uptake capacity of the group treated with Poria cocos B was significantly improved. The aforementioned results show that Poria cocos B can also regulate blood sugar by enhancing the ability of adipocytes to uptake glucose.

[0092] As shown in the above embodiments, the Poria cocos peel extract, as well as Poria cocos acid A and Poria cocos acid B of the present invention can indeed enhance the ability of biological cells to take up blood sugar, and can be used to regulate blood sugar, especially to reduce excessively high blood sugar.

Claims

1. A method for providing an extract from the peel of Poria cocos, characterized in that, The operation includes the following steps: (a) Extracting a piece of Poria cocos peel with a first solvent to obtain a crude extract; (b) Dry the crude extract to obtain a crude extract powder; and (c) Extract the crude extract powder with a second solvent to obtain a Poria cocos peel extract. The first solvent is an aqueous ethanol solution, and the second solvent is an aqueous ethanol solution or an alkaline solution. The Poria cocos peel extract contains pachymic acid A and pachymic acid B, but does not contain pachymic acid, dehydropachymic acid, or thomolic acid.

2. The method as described in claim 1, characterized in that, The first solvent and the second solvent are aqueous solutions of ethanol with the same or different concentrations.

3. The method as described in claim 1, characterized in that, The first solvent is an aqueous solution of ethanol and the second solvent is an alkaline solution.

4. The method as described in claim 3, characterized in that, Repeat step (c) with the same or different alkali solutions.

5. The method according to any one of claims 1 to 4, characterized in that, Prior to step (b), the skin of Poria cocos is repeatedly extracted with the same or different first solvents, and the extracts obtained from the multiple extractions are combined to provide the crude extract for step (b).

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