COMPOSITION CONTAINING GLUCORAPHANIN AND USE THEREOF

Abstract

The present disclosure relates to a composition comprising glucoraphanin and its therapeutic use.

Claims

1. A composition, comprising the following components: 1) component I providing glucoraphanin; 2) component II providing myrosinase; and 3) a basic salt compound.

2. The composition according to claim 1, further comprising ascorbic acid.

3. The composition according to claim 1, wherein the component I is selected from the group consisting of cruciferous plants, extracts thereof, and mixtures thereof.

4. The composition according to claim 1, wherein the cruciferous plant is broccoli.

5. The composition according to claim 3, wherein the cruciferous plant is whole of the plant, a part of the plant, or a mixture thereof.

6. The composition according to claim 3, wherein the cruciferous plant is selected from the group consisting of flower bulbs, seeds, sprouts and mixtures thereof.

7. The composition according to claim 1, wherein the basic salt compound is selected from the group consisting of sodium carbonate, potassium carbonate, sodium hydrogencarbonate, potassium hydrogencarbonate, sodium pyrophosphate, sodium citrate, potassium pyrophosphate, potassium citrate and mixtures thereof.

8. The composition according to claim 1, wherein the mass ratio of the component I, the component II and the basic salt compound is 10-80:1-80:0.1-1.

9. The composition according to claim 1 which is in form of solid.

10. The composition according to claim 1, which is in form of a preparation of powder, granule, capsule or tablet.

11. The composition according to claim 1, wherein the component I is selected from the group consisting of broccoli seed extract, broccoli sprout powder, broccoli flower bulb lyophilized powder, and mixtures thereof.

12. The composition according to claim 1, wherein the component II is selected from the group consisting of horseradish, radish, kale, a juice or a slurry thereof, and an extract thereof.

13. A method of preventing and/or treating a disease or disorder that can be prevented and/or treated by glucoraphanin, comprising administering the composition according to claim 1 to a subject in need thereof.

14. The method according to claim 13, wherein the disease or disorder that can be prevented and/or treated by glucoraphanin is selected from the group consisting of cancer, diabetes, cardiovascular diseases, Helicobacter pylori infection, autism, schizophrenia, depression, Alzheimer's disease and pulmonary fibrosis.

15. A method of converting glucoraphanin to sulforaphane in vitro, comprises the steps of: 1) providing a composition according to claim 1, and 2) mixing the composition with water or an aqueous solution.

16. A method of supplementing sulforaphane to a subject in need thereof, comprising administering to the subject a composition according to claim 1.

Description

DETAILED DESCRIPTION O F THE INVENTION

Example 1

[0033] (1) Preparation of composition 1 of the present invention: 720 g of broccoli seed aqueous extract (containing 13.0% of glucoraphanin, purchased from Brassica Protection Products LLC, the same as below) was mixed evenly with 296 g of horseradish powder and 10 g of sodium carbonate, to obtain 1.03 kg of Composition 1, wherein glucoraphanin accounted for 9.09%. The above composition 1 was placed in a sachet package in a weight of 5 g per bag to obtain a corresponding powder product.

[0034] (2) Preparation of control composition 1: 720 g of broccoli seed water extract (containing 13.0% of glucoraphanin) and 296 g of horseradish powder were mixed evenly to obtain 1.02 kg of the control composition 1, wherein glucoraphanin accounted for 9.18%. The above control composition 1 was placed in a sachet package in a weight of 5 g per bag to obtain a corresponding powder product.

[0035] (3) Accelerated stability experiment: The above two powder products were placed in an accelerated test chamber at 37° C. and 75% relative humidity for 3 months, and then taken out, and the appearance change was observed, and the sulforaphane production rate was measured by the following method.

[0036] 1.03 g of composition 1 and 1.02 g of control composition 1 (both containing glucoraphanin 93.6 mg) were taken, and added to 30 mL of water to incubate the simulated brewing conditions at 37° C. Samples was taken at 5 min, 8 min and 30 min respectively. The sulforaphane content was determined by HPLC and the sulforaphane production rate was calculated.

[0037] HPLC method for determination of sulforaphane: The sample solution was taken and passed through a 0.45 μm filter for HPLC analysis. HPLC conditions: column: Huapu Unitary C18 (4.6 mm×250 mm, 5 μm); column temperature: 30° C.; mobile phase: 70% water −30% acetonitrile; flow rate: 0.8 mL/min; injection volume: 10 μL; U V detection wavelength: 245 nm.

[0038] The experimental results are shown in Table 1 below.

TABLE-US-00001 TABLE 1 Comparison of sulforaphane production rates for Composition 1 and Control Composition 1 powder products sulforaphane production rate (%) 5 8 30 group min min min appearance taste Composition 1 26 38 50 light yellow own favour (month 0) powder Composition 1 16 25 45 light yellow own favour (month 3) powder Control 26 38 49 light yellow own favour composition 1 powder (month 0) Control 10 19 31 pink powder with composition 1 obvious (month 3) bitterness

[0039] It can be seen from the experimental results that the composition 1 of the present invention containing the basic salt has no significant change in appearance and taste after the accelerated experiment, and the sulforaphane production rate is kept better than that of the control composition 1 containing no basic salt. The rate of sulforaphane production was basically unchanged before and after the accelerated experiment, which indicated that the sulforaphane content in the composition after hydrolysis was almost unchanged from that before hydrolysis.

Example 2

[0040] (1) Preparation of tablets 1 of the present invention: 200 g of broccoli seedling aqueous extract (containing 13.0% of glucoraphanin), 200 g of horseradish powder, 4 g of vitamin C, 10 g of sodium phosphate and 596 g of tablet excipients (consisting of starch, maltodextrin, and hydroxypropylmethylcellulose, with a ratio (w/w) of 5:80:2, the same below) were evenly mixed, and were tableted based on 0.6 g per tablet, covered with film coating to give 1.02 kg of the tablets 1 of the invention, wherein glucoraphanin accounted for 2.57%. The above-mentioned tablets 1 of the present invention were placed in a bottle with 60 tablets per bottle and a desiccant was added thereto, and the bottle was sealed to obtain a corresponding tablet product.

[0041] (2) Preparation of Control Tablet 1: 200 g of aqueous extract of broccoli seedling (containing 13.0% of glucosinolate), 200 g of horseradish powder, 4 g of vitamin C and 596 g of tablet excipients were evenly mixed, and were tableted based on 0.6 g per tablet, covered with film coating to give 1.01 kg of Control Tablet 1, of which glucoraphanin accounted for 2.6%. The above-mentioned tablet 1 of the present invention was placed in a bottle with 60 tablets per bottle and a desiccant was added thereto, and the bottle was sealed to obtain a corresponding tablet product.

[0042] (3) Accelerated stability experiment: The above two tablet products were placed in an accelerated test chamber at 37° C. and 75% relative humidity for 3 months, and then taken out, and the appearance change was observed and the sulforaphane production rate was measured by the following method.

[0043] 30 g of each tablet sample was taken and ground. 1.02 g of the tablet 1 powder of the invention and 1.01 g of the control tablet 1 powder (both containing 26 mg of glucoraphanin) were taken, respectively, and added to a 30 mL artificial postprandial gastric juice simulating solution (the artificial postprandial gastric juice simulating solution was prepared according to Chinese Pharmacopoeia, Part II, 2015 edition, and the pH was adjusted to 3.5), and the temperature was maintained at 37° C. The samples were taken at 30 min and 60 min respectively. The sulforaphane content was determined by HPLC to calculate the sulforaphane production rate. The HPLC method for the determination of sulforaphane was the same as in Example 1.

[0044] The experimental results are shown in Table 2 below.

TABLE-US-00002 TABLE 2 Comparison of sulforaphane production rates for Table 1 and Control Table 1 sulforaphane production rate (%) group 30 min 60 min appearance taste Tablet 1 of the 17.5 23.0 light white own favour present invention tablet (month 0) Tablet 1 of the 11.1 16.4 light white own favour present invention tablet (month 3) Control tablet 1 17.6 23.3 light white own favour (month 0) tablet Control tablet 1 7.9 11.9 pink tablet with (month 3) obvious bitterness

[0045] It can be seen from the experimental results that the tablet 1 of the present invention containing the basic salt has no significant change in appearance and taste after the accelerated experiment, and the rate of sulforaphane production remains well compared to the control tablet 1 containing no basic salt. After the accelerated stability experiment, the rate of sulforaphane production was significantly higher than that of the control tablets.

Example 3

[0046] (1) Preparation of tablets 2 of the present invention: 200 g of broccoli seed aqueous extract (containing 13.0% of glucoraphanin), 50 g broccoli seedling powder (containing 4.5% of glucoraphanin), 50 g broccoli flower bulb lyophilized powder, 250 g horseradish powder, 4 g of calcium vitamin C, 10 g of sodium citrate and 596 g of tablet excipients (consisting of starch, maltodextrin, and hydroxypropylmethylcellulose, with a ratio (w/w) of 5:80:2) were evenly mixed, and were tableted based on 0.6 g per tablet, covered with film coating to give 1.16 kg of the tablets 2 of the invention, wherein glucoraphanin accounted for 2.80%. The above-mentioned tablets 2 of the present invention were placed in a bottle with 60 tablets per bottle and a desiccant was added thereto, and the bottle was sealed to obtain a corresponding tablet product.

[0047] (2) Preparation of Control Tablet 2: 200 g of aqueous extract of broccoli seeds (containing 13.0% of glucosinolate), 50 g broccoli seedling powder (containing 4.5% of glucoraphanin), 50 g broccoli flower bulb lyophilized powder, 250 g horseradish powder, 4 g of calcium vitamin C, 10 g of sodium phosphate and 596 g of tablet excipients (consisting of starch, maltodextrin, and hydroxypropylmethylcellulose, with a ratio (w/w) of 5:80:2) were evenly mixed, and were tableted based on 0.6 g per tablet, covered with film coating to give 1.15 kg of the control tablets 2 of the invention, wherein glucoraphanin accounted for 2.83%. The above-mentioned control tablets 2 of the present invention were placed in a bottle with 60 tablets per bottle and a desiccant was added thereto, and the bottle was sealed to obtain a corresponding control tablet product.

[0048] (3) Accelerated stability experiment: The above two tablet products were placed in an accelerated test chamber at 37° C. and 75% relative humidity for 3 months, and then taken out, and the appearance change was observed and the sulforaphane production rate was measured by the following method.

[0049] 100 g of each tablet sample was taken and ground. 1.16 g of the tablet 2 powder of the invention and 1.15 g of the control tablet 2 powder (both containing 28 mg of glucoraphanin) were taken, respectively, and added to a 30 mL artificial postprandial gastric juice simulating solution (the artificial postprandial gastric juice simulating solution was prepared according to Chinese Pharmacopoeia, Part II, 2015 edition, and the pH was adjusted to 3.5), and the temperature was maintained at 37° C. The samples were taken at 30 min and 60 min respectively. The sulforaphane content was determined by HPLC to calculate the sulforaphane production rate. The HPLC method for the determination of sulforaphane was the same as in Example 1.

[0050] The experimental results are shown in Table 3 below.

TABLE-US-00003 TABLE 3 Comparison of sulforaphane production rates for Table 2 and Control Table 2 sulforaphane production rate (%) group 30 min 60 min appearance taste Tablet 2 of the 24.7 38.2 light white own favour present invention tablet (month 0) Tablet 2 of the 14.1 22.6 light white own favour present invention tablet (month 3) Control tablet 2 24.7 38.2 light white own favour (month 0) tablet Control tablet 2 9.2 15.3 pink tablet with (month 3) obvious bitterness

[0051] It can be seen from the experimental results that the tablet 2 of the present invention containing the basic salt has no significant change in appearance and taste after the accelerated experiment, and the rate of sulforaphane production remains well compared to the control tablet 2 containing no basic salt. After the accelerated stability experiment, the rate of sulforaphane production was significantly higher than that of the control tablets.