阅读说明：本技术 用于管理高糖血症及相关病况的组合物 (Compositions for managing hyperglycemia and related conditions ) 是由 M.马杰德 K.纳加布休沙纳姆 L.蒙德库尔 于 2018-12-26 设计创作，主要内容包括：公开了一种使用含有百里氢醌的组合物治疗性管理哺乳动物中高血糖的方法。更具体地,本发明公开了含有百里氢醌的组合物,其用于抑制酶α-葡糖苷酶的活性并增加哺乳动物细胞对葡萄糖的细胞摄取。在本文中也公开了百里氢醌的抗氧化、抗炎和抗糖化作用。(A method of therapeutically managing hyperglycemia in a mammal using a thymoquinol-containing composition is disclosed. More specifically, the present invention discloses compositions containing thymohydroquinone for inhibiting the activity of the enzyme alpha-glucosidase and increasing the cellular uptake of glucose by mammalian cells. Antioxidant, anti-inflammatory and anti-glycation effects of thymohydroquinone are also disclosed herein.)

1. A method of inhibiting glucosidase comprising the steps of:

i) contacting a glucosidase with a p-nitrophenyl-alpha-d-glucopyranoside substrate;

ii) incubating under optimal conditions with an effective dose of thymoquinol or a composition comprising thymoquinol;

iii) reading the change in absorbance using spectrophotometry and fluorimetry;

iv) comparing the absorbance to a control blank and determining the percent enzyme Inhibition (IC) of thymohydroquinone or a composition comprising thymohydroquinone using the formula₅₀)：

% inhibition ═ control absorbance-inhibitor absorbance/control absorbance ] x 100.

2. The method of claim 1, wherein the thymoquinone-comprising composition comprises about 0.1-5% w/w thymoquinone, about 0.01-10% w/w thymohydroquinone, about 20-95% w/w fatty acids, about 0.001-3% w/w alpha-hederagenin or hederagenin, 0.1-4.0% w/w stabilizer, and 0.2-2% w/w bioavailability enhancer.

3. The composition of claim 2, wherein the stabilizer is selected from the group comprising: rhodanolic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherol.

4. The composition according to claim 2, wherein the bioavailability enhancing agent is selected from the group comprising: piperine, quercetin, garlic extract, ginger extract and naringin.

5. A method of increasing glucose uptake in a mammalian cell, the method comprising the steps of: contacting mammalian cells with an effective dose of thymohydroquinone or a composition comprising thymohydroquinone to increase glucose uptake by said cells.

6. The method of claim 5, wherein the composition comprises about 0.1-5% w/w thymoquinone, about 0.01-10% w/w thymohydroquinone, about 20-95% w/w fatty acids, about 0.001-3% w/w alpha-hederagenin or hederagenin, 0.1-4.0% w/w stabilizer, and 0.2-2% w/w bioavailability enhancer.

7. The composition of claim 6, wherein the stabilizer is selected from the group comprising: rhodanolic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherol.

8. The composition according to claim 6, wherein the bioavailability enhancing agent is selected from the group comprising: piperine, quercetin, garlic extract, ginger extract and naringin.

9. The composition of claim 6, wherein the mammalian cell is a human cell.

10. A method for therapeutically managing hyperglycemia and related conditions in a mammal, the method comprising the steps of: administering an effective dose of thymoquinol or a composition comprising thymoquinol to lower the glucose level in blood.

11. The method of claim 10, wherein glucose absorption is reduced by inhibiting glucosidase, increasing cellular uptake of glucose, reducing free radicals, reducing inflammation, and reducing glycation, thereby causing management of hyperglycemia and related conditions.

12. The method of claim 10, wherein the hyperglycemia-related condition is present in a disease state selected from the group comprising: diabetes, obesity, hyperlipoproteinemia, hyperlipidemia, cardiovascular complications, cancer, atherosclerosis, neurodegenerative diseases, allergy, inflammation and osteoporosis.

13. The method of claim 10, wherein the composition comprises about 0.1-5% w/w thymoquinone, about 0.01-10% w/w thymohydroquinone, about 20-95% w/w fatty acids, about 0.001-3% w/w alpha-hederagenin or hederagenin, 0.1-4.0% w/w stabilizer, and 0.2-2% w/w bioavailability enhancer.

14. The composition of claim 13, wherein the stabilizer is selected from the group comprising: rhodanolic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherol.

15. The composition according to claim 13, wherein the bioavailability enhancing agent is selected from the group comprising: piperine, quercetin, garlic extract, ginger extract and naringin.

16. The method of claim 10, wherein the mammalian cell is a human cell.

17. The composition of claim 13, wherein the composition is formulated with a pharmaceutically/nutraceutically acceptable excipient, adjuvant, diluent, or carrier and administered orally in the form of a tablet, capsule, soft gel, syrup, fondant, powder, suspension, emulsion, chew, candy, or food.

Technical Field

The present invention relates to the therapeutic management of hyperglycemia in mammals. More particularly, the present invention relates to compositions comprising thymohydroquinone and their therapeutic potential in the management of hyperglycemia and its associated conditions.

Background

Summary of The Invention

The present invention discloses novel compositions comprising thymohydroquinone. In particular, compositions containing thymohydroquinone for inhibiting the activity of the enzyme alpha-glucosidase are disclosed. The invention also discloses the use of a composition comprising thymoquinol for increasing the cellular uptake of glucose by mammalian cells. More specifically, the present invention discloses methods of therapeutically managing hyperglycemia in mammals using compositions containing thymoquinol. Antioxidant, anti-inflammatory and anti-glycation effects of thymohydroquinone are also disclosed herein.

Other features and advantages of the present invention will become apparent from the following more detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, the principles of the invention.

Brief Description of Drawings

FIGS. 1a and 1b show graphical representations of alpha-glucosidase inhibitory activity of Thymoquinone (TQ), Thymoquinol (THQ), and thymoquinol compositions.

Figure 2 is a graph showing the percent glucose uptake by adipocytes and muscle cells treated with Thymoquinone (TQ), Thymoquinol (THQ), and thymoquinol compositions.

Figure 3 shows representative histograms of glucose uptake by mammalian cells treated with insulin (3a), Thymoquinone (TQ) (3b), Thymoquinol (THQ) (3c) and thymoquinol composition (3 d). Untreated cells served as a control group (3e),

FIG. 4 shows a graphical representation of DPPH scavenging activity of Thymoquinone (TQ) (4a), Thymohydroquinone (THQ) (4b) and thymohydroquinone composition (4 c).

Figure 5 shows a graphical representation of DPPH scavenging activity and glucosidase inhibitory activity compositions as the percentage of thymoquinol increases. An increase in thymohydroquinone content is directly related to an increase in biological activity.

Figure 6 shows a graphical representation of DPPH scavenging activity and glucosidase inhibitory activity compositions as the percentage of thymoquinone increases. The increase in thymoquinone content is inversely proportional to the increase in biological activity.

Description of the most preferred embodiments

In a most preferred embodiment, the present invention discloses a method of inhibiting glucosidase comprising the steps of:

i) contacting a glucosidase with a p-nitrophenyl-alpha-d-glucopyranoside substrate;

ii) incubating under optimal conditions with an effective dose of thymoquinol or a composition comprising thymoquinol;

iii) reading the change in absorbance using spectrophotometry and fluorimetry;

iv) comparing the absorbance to a control blank and determining the percent enzyme Inhibition (IC) of thymohydroquinone or a composition comprising thymohydroquinone using the formula₅₀)：

% inhibition ═ control absorbance-inhibitor absorbance/control absorbance ] x 100.

In a related embodiment, the composition comprises about 0.1% -5% w/w thymoquinone, about 0.01% -10% w/w thymoquinol, about 20% -95% w/w fatty acids, about 0.001% -3% w/w alpha-hederagenin or hederagenin, 0.1% -4.0% w/w stabilizer, and 0.2% -2% w/w bioavailability enhancer. In another related embodiment, the stabilizing agent is selected from the group comprising: rhodanolic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherol. In another related embodiment, the bioavailability enhancing agent is selected from the group comprising: piperine, quercetin, garlic extract, ginger extract, and naringin (naringin).

In another most preferred embodiment, the present invention discloses a method of increasing glucose uptake in a mammalian cell, said method comprising the steps of: contacting mammalian cells with an effective dose of thymohydroquinone or a composition comprising thymohydroquinone to increase glucose uptake by said cells. In a related embodiment, the composition comprises about 0.1% -5% w/w thymoquinone, about 0.01% -10% w/w thymoquinol, about 20% -95% w/w fatty acids, about 0.001% -3% w/w alpha-hederagenin or hederagenin, 0.1% -4.0% w/w stabilizer, and 0.2% -2% w/w bioavailability enhancer. In another related embodiment, the stabilizing agent is selected from the group comprising: rhodanolic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherol. In another related embodiment, the bioavailability enhancing agent is selected from the group comprising: piperine, quercetin, garlic extract, ginger extract and naringin. In another related embodiment, the mammalian cell is a human cell.

In another preferred embodiment, the present invention discloses a method for the therapeutic management of hyperglycemia and related conditions in a mammal, comprising the steps of: administering an effective dose of thymoquinol or a composition comprising thymoquinol to lower the glucose level in blood. In related embodiments, glucose absorption is reduced by inhibiting glucosidase, increasing cellular uptake of glucose, reducing free radicals, reducing inflammation, and reducing glycation, thereby causing control of hyperglycemia and related conditions. In another related embodiment, the hyperglycemia-related condition is present in a disease state selected from the group consisting of: diabetes, obesity, hyperlipoproteinemia, hyperlipidemia, cardiovascular complications, cancer, atherosclerosis, neurodegenerative diseases, allergy, inflammation and osteoporosis. In a related embodiment, the composition comprises about 0.1% -5% w/w thymoquinone, about 0.01% -10% w/w thymohydroquinone, about 20% -95% w/w fatty acids, about 0.001% -3% w/w alpha-hederagenin or hederagenin, 0.1% -4.0% w/w stabilizer, and 0.2% -2% w/w bioavailability enhancer. In another related embodiment, the stabilizing agent is selected from the group comprising: rhodanolic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherol. In another related embodiment, the bioavailability enhancing agent is selected from the group comprising: piperine, quercetin, garlic extract, ginger extract and naringin. In another related embodiment, the mammalian cell is a human cell. In another related embodiment, the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally in the form of tablets, capsules, soft gels, syrups, gummies, powders, suspensions, emulsions, chews, candies or foods.

The above-described most preferred embodiments incorporating the technical features and technical effects of the present invention are illustrated by the following illustrative examples.

Example 1: inhibition of glucosidase

To inhibit glucosidase, α -glucosidase (code G5003; Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 67mM potassium phosphate buffer, pH 6.8, containing 8, containing 0.2% bovine serum albumin (Sigma-Aldrich) and 0.02% sodium azide (Sigma-Aldrich) as the enzyme source, thymoquinone, and a composition containing thymoquinol were weighed, prepared at 63, 125, 250, and 500 μ G/ml, and made up with an equal volume of distilled water, 50 μ l of the composition was incubated with 50 μ l of the enzyme source (0.15U/ml) for 5 minutes, after which 50 μ l of the substrate (1.25mM) was added, and at room temperature for 20 minutes, before and after addition of substrate, the absorbance was measured on a microplate reader (BMG UOstar TIMA microplate reader) at 405nm and the difference in absorbance was measured as the difference between the absorbance of the negative inhibition of the enzyme, that was measured as the difference in absorbance of absorbance, and the absorbance was calculated as the difference between the absorbance of the absorbance measured for each of the absorbance, namely the absorbance of the absorbance measured as the difference between the absorbance of the control solution, where the absorbance was calculated as the absorbance, and the absorbance, where the absorbance was calculated as the absorbance, where the difference was calculated as the absorbance was 100₅₀Value (i.e. the concentration of the composition that results in 50% inhibition of the maximal activity).

In combination with thymoquinone (IC)₅₀407.6 μ g/ml) relative to each other, thymoquinol (IC)₅₀71.9. mu.g/ml) and thymohydroquinone (IC)₅₀150.9 μ g/ml) showed effective inhibition of α glucosidase (fig. 1a and 1 b).