Traditional chinese medicine, extract and composition thereof for reducing uric acid

Abstract

The application discloses a Stenzel's cymbalaria, a Stenzel's cymbalaria extract or a composition containing the Stenzel's cymbalaria or the extract thereof, and a use of the above products in preparation of medicines, health products, foods or food additives for reducing uric acid. The above products have a remarkable effect of reducing uric acid and small side effects, and the preparation method is simple and efficient, and is suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates generally to the field of traditional Chinese medicine, and in particular to a traditional Chinese medicine for lowering uric acid, an extract and a composition thereof, and a preparation method. BACKGROUND

[0002] Abnormal elevation of uric acid is mainly caused by excessive production of uric acid or decreased excretion of uric acid. If the blood uric acid in the human body is at a high level for a long time, it will form hyperuricemia, further develop into gout, and cause the occurrence of diseases such as coronary heart disease, hypertension, diabetes, and chronic kidney disease, which seriously endanger people's health.

[0003] Traditional Chinese medicine resources are an organic part of natural resources, among which medicinal plants belong to renewable resources. With the development of China's national economy and traditional Chinese medicine industry, the demand for traditional Chinese medicinal materials continues to increase, making traditional Chinese medicine resources face more severe challenges. Expanding new drug sources from ethnic folk herbal resources and enriching new resource sources of traditional Chinese medicine have become one of the effective strategies to ensure the sustainable use of traditional Chinese medicine resources and the overall requirement of "total amount not reduced and supply guaranteed".

[0004] Pteridium aquilinum var. obliquum is a selenium-rich resource with both medicinal and edible characteristics in Enshi Tujia and Miao Autonomous Prefecture, Hubei Province, and is a kind of hyper-selenium plant. Pteridium aquilinum var. obliquum is distributed in Hubei and Hunan provinces, and has a long history of being used as food in Hunan and Hubei areas. It also has potential pharmacological activity. According to the Chinese Plant Flora, the whole plant can be used as a wild vegetable and also has medicinal effects, has the effect of clearing heat and removing dampness, and has very good potential pharmacological activity. It has the effects of clearing heat and removing dampness, promoting water and reducing swelling.

[0005] According to traditional Chinese medicine, the elevation of blood uric acid is caused by the habit of eating fatty and thick food, the accumulation of phlegm, the accumulation of phlegm, and the downward flow of dampness and heat. The most common syndrome in patients is the syndrome of dampness and heat. Tracing the medicinal history of Pteridium aquilinum var. obliquum, it has the effects of clearing heat and removing dampness, promoting water and reducing swelling, which is exactly the etiology and pathogenesis of the elevation of blood uric acid.

[0006] The problem to be solved at present is to find and verify a medicinal and edible resource with the effect of lowering uric acid by using modern scientific and technological equipment and methods. SUMMARY

[0007] One or more embodiments of the present application provide a Pteridium aquilinum var. obliquum water extract, which has a total flavonoid content of 0.15-0.2% by weight, for example, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2% by weight.

[0008] One or more embodiments of the present application provide a composition with uric acid-lowering effect, comprising Pteroxygonum giraldii or a water extract thereof, wherein the total flavonoid content of the Pteroxygonum giraldii is 0.5-0.6% by weight, for example 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, or 0.6% by weight; and the total flavonoid content of the Pteroxygonum giraldii water extract is 0.1-0.2% by weight, for example 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, or 0.2% by weight.

[0009] In one or more embodiments, the composition comprises an adjuvant or an excipient.

[0010] In one or more embodiments, the adjuvant is dextrin and / or sucralose.

[0011] In one or more embodiments, the composition is prepared into a decoction, a tea, granules, pills, powder, tablets, capsules, or oral liquid.

[0012] One or more embodiments of the present application provide a preparation method of the Pteroxygonum giraldii water extract of the present application, comprising: taking Pteroxygonum giraldii, soaking and decocting with water, concentrating, to obtain the Pteroxygonum giraldii water extract.

[0013] In one or more embodiments, in the preparation method, the Pteroxygonum giraldii is soaked with water for 30-40 minutes (for example 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 minutes), decocted twice, with the decocting time being 30-40 minutes (for example 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 minutes) and 20-30 minutes (for example 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 minutes) respectively, the volume of the water is 18-22 mL / g (for example 18, 19, 20, 21, or 22 mL / g) and 8-12 mL / g (for example 8, 9, 10, 11, or 12 mL / g) respectively, the weight of the medicinal material, and the filtrates of the two decoctions are combined, concentrated, to obtain the Pteroxygonum giraldii water extract.

[0014] In one or more embodiments, in the preparation method, the filtrate is concentrated at 55-65℃ (55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65℃).

[0015] One or more embodiments of the present application provide the use of Pteroxygonum giraldii, the Pteroxygonum giraldii water extract of the present application, or the composition in the preparation of a medicine, health product, food, or food additive for lowering uric acid.

[0016] One or more embodiments of the present application provide use of Pteroxygonum giraldii, a Pteroxygonum giraldii water extract or a composition of the present application in the preparation of a medicament or health product for preventing and / or treating a disease associated with high uric acid.

[0017] In one or more embodiments, the disease associated with high uric acid is uric acidemia, hyperuricemic nephritis, hyperuricemic hypertension, hyperuricemic heart disease, gout kidney stones or gout. DETAILED DESCRIPTION

[0018] In one or more embodiments, the method for preparing a Pteroxygonum giraldii water extract comprises the following steps:

[0019] Step a: Pteroxygonum giraldii is decocted in water twice, the extract is combined and filtered, and then prepared for use;

[0020] Step b: The extract obtained in step a is concentrated to an appropriate concentration at 55-65°C to obtain the water extract.

[0021] In one or more embodiments, the decoction time in step a is 30 minutes and 20 minutes, respectively, the volume of water used in the first decoction is 18-22 times the weight of the medicinal material (mL / g), and the volume of water used in the second decoction is 8-12 times the weight of the medicinal material (mL / g).

[0022] In one or more embodiments, the method for preparing a Pteroxygonum giraldii water extract comprises the following steps:

[0023] Step a: Pteroxygonum giraldii is powdered and prepared for use;

[0024] Step b: The powder obtained in step a is passed through a 10-60 mesh sieve (e.g., 10, 20, 30, 40, 50 or 60 mesh);

[0025] Step c: The powder obtained in step b is packaged to obtain the water extract (which can be used as a tea).

[0026] In one or more embodiments, the method for preparing a Pteroxygonum giraldii water extract comprises the following steps:

[0027] Step a: Pteroxygonum giraldii is decocted in water twice, the extract is combined and filtered, and then prepared for use;

[0028] Step b: The above clear paste is mixed with excipients to form granules, which are then filled to obtain granules of the water extract.

[0029] In one or more embodiments, the two times of decocting in step a are 30 minutes and 20 minutes, respectively, the volume of water used in the first time of decocting is 18-22 times the weight of the medicinal material, and the volume of water used in the second time of decocting is 8-12 times the weight of the medicinal material.

[0030] In one or more embodiments, the drying method in step a is reduced pressure drying, and the auxiliary materials used are dextrin and sucralose.

[0031] In one or more embodiments, the medicinal part of Cardamine violifolia O.E. Schuiz is the stems and leaves of the perennial herb Cardamine violifolia O.E. Schuiz.

[0032] One or more embodiments of the present application also provide an alcohol extract of Cardamine violifolia O.E. Schuiz.

[0033] One or more embodiments of the present application also provide an alcohol extract of Cardamine violifolia O.E. Schuiz.

[0034] In one or more embodiments, Cardamine violifolia O.E. Schuiz is extracted with ethanol, the extract solutions are combined, and concentrated to obtain an alcohol extract of Cardamine violifolia O.E. Schuiz.

[0035] In one or more embodiments, the present application at least comprises at least one of the following beneficial effects:

[0036] The effect of lowering uric acid is significant, the side effects are small, and the patient's kidney function is protected at the same time, which has great significance in preventing and treating high uric acid related diseases;

[0037] For the pathogenesis of damp-heat with elevated blood uric acid, it can be applied in the prevention and treatment of high uric acid related diseases and health products;

[0038] It is a dual-purpose resource of medicine and food, and has the advantages of significant curative effect, small side effect, convenient taking, and economic price;

[0039] The production process is simple and efficient, and is conducive to industrialized production;

[0040] The single medicine has the advantages of few ingredients and simple preparation, and is suitable for industrialized production.

[0041] Examples

[0042] The present application will be further described in detail below in combination with examples. It should be noted that, in the following examples, the experimental methods are conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial channels unless otherwise specified.

[0043] Example 1

[0044] Step a: the water decocting 2 times, respectively, and then combine the extract, filter, and reserve for later use.

[0045] Step b: the extract obtained in step a is concentrated to a proper concentration at 55-65℃, and then the decoction of the dual-purpose resource of medicine and food is obtained.

[0046] Example 2

[0047] Step a: the water decocting 2 times, respectively, and then combine the extract, filter, and reserve for later use.

[0048] Step b: the powder obtained in step a is passed through a 10-60 mesh sieve.

[0049] Step c: the powder obtained in step b is bagged, and then the tea of the dual-purpose resource of medicine and food is obtained.

[0050] Example 3

[0051] Step a: the water decocting 2 times, respectively, and then combine the extract, filter, and reserve for later use.

[0052] Step b: the above clear paste is mixed with excipients to form granules, and then the granules are filled, and then the granules of the traditional Chinese medicine composition are obtained.

[0053] Example 4: determination of the total flavonoids content of P. cordata

[0054] 1. Instruments and reagents

[0055] 1.1 Reagents

[0056] Sodium nitrite (NaNO2), aluminum nitrate (Al(NO3)3·9H2O), sodium hydroxide (NaOH), petroleum ether (60-90℃), anhydrous ethanol (CH3CH2OH), methanol (CH3OH), and polyamide powder.

[0057] 1.2 Preparation of reagents

[0058] 5% sodium nitrite solution: weigh 0.5g of sodium nitrite, dissolve in water to 10mL.

[0059] 10% aluminum nitrate solution: weigh 1.76g of aluminum nitrate, dissolve in water to 10mL.

[0060] Sodium hydroxide test solution: weigh 1.075g of sodium hydroxide, dissolve in water to 25mL.

[0061] 1.3 Instruments

[0062] UV / visible spectrophotometer, ultrasonic cleaner, centrifuge, No.4 sieve, analytical balance (sensitivity of 0.01mg, 0.0001g and 0.001g, respectively), and chromatography column.

[0063] 2. Determination of total flavonoids content

[0064] Determination of preparation method of test solution

[0065] The test sample was prepared by grinding and sieving the original powder of P. cordata. 1 g of the powder was precisely added to 25 mL of methanol, and ultrasonicated for 30 min. The supernatant was obtained by centrifugation, and the test solution was prepared. 3.0 mL of the supernatant was taken and adsorbed on 1 g of polyamide powder in an evaporating dish. The methanol was removed by water bath evaporation, and then transferred to a chromatographic column. The column was first eluted with 20 mL of dichloromethane, and the dichloromethane solution was discarded. Then the column was eluted with methanol, and the eluate was combined, evaporated to dryness by water bath, and dissolved in 10 mL of water.

[0066] 6.0 mL of the test solution was precisely taken into a 25 mL volumetric flask, and water was added to 6 mL. 1 mL of 5% sodium nitrite solution was added, and the mixture was shaken and allowed to stand for 6 min. 1 mL of 10% aluminum nitrate solution was added, and the mixture was shaken and allowed to stand for 6 min. 10 mL of sodium hydroxide solution was added, and the mixture was shaken and allowed to stand for 15 min. The absorbance value (A) was measured at a wavelength of 510 nm, with 0.0 mL of the control solution as the blank.

[0067] Determination of maximum absorption wavelength

[0068] 10.05 mg of rutin control was precisely weighed, dissolved in methanol, and made up to 50 mL to prepare a rutin standard stock solution with a concentration of 0.201 mg / mL. 2 mL of the rutin control was precisely taken into a 25 mL volumetric flask, and 1 mL of 5% sodium nitrite solution was added. The mixture was shaken and allowed to stand for 6 min. 1 mL of 10% aluminum nitrate solution was added, and the mixture was shaken and allowed to stand for 6 min. 10 mL of sodium hydroxide solution was added, and the mixture was shaken and allowed to stand for 15 min. The wavelength was scanned from 200 nm to 800 nm, and the maximum absorption was found at 510 nm. Therefore, the detection wavelength was determined to be 510 nm.

[0069] 3. Determination of total flavonoids content

[0070] Sample weight: 1.0000 g

[0071] Total flavonoids content of P. cordata:

[0072] X = content of total flavonoids in the sample, calculated as rutin (C 27 H 30 O 16 ) and expressed in grams per 100 grams or grams per 100 milliliters (g / 100 g or g / 100 mL);

[0073] C - Concentration of total flavonoids in the test sample solution, unit: mg / mL;

[0074] V1 - Volume of the test sample, unit: mL;

[0075] V2 - Volume of the test sample solution, unit: mL;

[0076] V3 - Volume of the color developing solution, unit: mL;

[0077] V4 - Volume of the test sample after the water bath evaporation of methanol, unit: mL;

[0078] V5 - Volume of the test sample added in the polyamide powder, unit: mL;

[0079] M - Sample amount, unit: g or mL

[0080] After calculation, we get

[0081] The total flavonoids content of P. cordata is 0.590%.

[0082] Example 5 Determination of the flavonoids content of P. cordata water extract

[0083] 1. Instruments and reagents

[0084] 1.1 Reagents

[0085] Sodium nitrite (NaNO2), aluminum nitrate (Al (NO3) 3 9H2O), sodium hydroxide (NaOH), petroleum ether (60-90°C), anhydrous ethanol (CH3CH2OH), methanol (CH3OH).

[0086] 1.2 Preparation of reagents

[0087] 5% sodium nitrite solution: weigh 0.5 g of sodium nitrite and dissolve in 10 mL of water.

[0088] 10% aluminum nitrate solution: weigh 1.76 g of aluminum nitrate and dissolve in 10 mL of water.

[0089] Sodium hydroxide test solution: weigh 1.075 g of sodium hydroxide and dissolve in 25 mL of water.

[0090] 1.3 Instruments

[0091] UV / visible spectrophotometer, ultrasonic cleaner, centrifuge, analytical balance (sensitivity of 0.01 mg, 0.0001 g and 0.001 g respectively)

[0092] 2. Sample processing method

[0093] Take 50.71 g of Pteridium aquilinum, weigh the amount as M, soak in water for 30 min, and extract twice. The first time, add 30 times the amount of water, boil, then keep boiling at medium heat for 30 min, filter while hot, and then add 20 times the amount of water to the residue, boil, then keep boiling at medium heat for 20 min, filter while hot. Combine the filtrates.

[0094] 3. Establishment of the method for determining the extract yield

[0095] Concentrate the filtrate in a water bath, and dry under reduced pressure (-0.07 MPa, 60°C) until the weight is constant. The weight of the dry extract powder is m.

[0096] The extract yield is m / M x 100%.

[0097] 4. Establishment of the method for determining the total flavone content

[0098] 4.1 Determination of the method for preparing the test solution

[0099] Take 1 g of the powder, precisely add 25 mL of methanol, and ultrasonicate for 30 min. Centrifuge, take the supernatant, and obtain the test solution. Take 4.0 mL of the supernatant, add 1 g of polyamide powder in an evaporating dish, evaporate the methanol in a water bath, then transfer into a chromatographic column. First, elute with 20 mL of dichloromethane, and discard the dichloromethane solution. Then, elute with methanol, combine the eluate, evaporate in a water bath, and make up to 10 mL to obtain the product.

[0100] Precisely take 0, 6.0 mL of the test solution into a 25 mL volumetric flask, add water to 6 mL, add 1 mL of 5% sodium nitrite solution, shake well, stand for 6 min, add 1 mL of 10% aluminum nitrate solution, shake well, stand for 6 min, add 10 mL of sodium hydroxide solution, shake well, add water to the mark, shake well, stand for 15 min, and determine the absorbance value (A) at a wavelength of 510 nm, with 0.0 mL of the solvent prepared from the control solution as the blank.

[0101] 4.2 Determination of the maximum absorption wavelength

[0102] Precisely weigh 10.05 mg of rutin control, dissolve in methanol, and make up to 50 mL to obtain a rutin standard stock solution with a concentration of 0.201 mg / mL. Precisely take 2 mL of the rutin control into a 25 mL volumetric flask, add 1 mL of 5% sodium nitrite solution, shake well, stand for 6 min, add 1 mL of 10% aluminum nitrate solution, shake well, stand for 6 min, add 10 mL of sodium hydroxide solution, shake well, add water to the mark, shake well, stand for 15 min, and scan the wavelength from 200 to 800 nm, with 0.0 mL of the control solution as the blank. The maximum absorption is found at 510 nm, so the detection wavelength is determined to be 510 nm.

[0103] 5. Determination of the total flavone content

[0104] Sample weight: 1.0180 g

[0105] Volume flask before ultrasonic: 47.5371 g

[0106] Volume flask after ultrasonic: 47.5336 g

[0107] Supplementary weight: 0.0035 g

[0108] Total flavonoids content of P. cordata

[0109] X—Total flavonoids content in the sample, calculated as rutin (C 27 H 30 O 16 ), in grams per 100 grams or grams per 100 milliliters (g / 100g or g / 100mL);

[0110] C—The concentration of total flavonoids in the test solution, read from the standard curve, in milligrams per milliliter (mg / mL);

[0111] V1—The sample volume, in milliliters (mL);

[0112] V2—The volume of the sample solution, in milliliters (mL);

[0113] V3—The color development volume, in milliliters (mL);

[0114] V4—The volume after the water bath evaporation of methanol (mL);

[0115] V5—The volume of the sample added to the polyamide powder (mL);

[0116] M—The sample weight, in grams or milliliters (g or mL)

[0117] After calculation, we get

[0118] The total flavonoids content of the water extract of P. cordata is 0.161%.

[0119] Example 6 Effect of P. cordata on uric acid, creatinine, urea nitrogen, and body weight levels in high uric acid model rats

[0120] Experimental principle and purpose: Potassium oxonate is a commonly used modeling drug for high uric acid models, which can simulate the state of elevated blood uric acid in humans. In this experiment, a high uric acid model of rats was used to evaluate the effect of P. cordata on uric acid, creatinine, urea nitrogen, and body weight levels in rats.

[0121] Experimental method: 48 SD male rats were adapted for 7 days, then fasted for 12 hours, and blood was taken from the eye orbit to detect serum uric acid value. 48 rats were randomly divided into: blank group, model group, positive drug group, high dose group, middle dose group, low dose group of the decoction prepared in Example 1, 8 rats in each group.

[0122] Blank group: intragastrically administered with the same solvent at the same dose every morning (7:00), and the other groups were intragastrically administered with potassium oxonate 1000 mg / kg every morning (7:00).

[0123] Blank group and model group: intragastrically administered with the same solvent every afternoon (16:00).

[0124] Positive drug group: intragastrically administered with allopurinol 27 mg / kg every afternoon (16:00).

[0125] Test drug group: intragastrically administered with the test sample diluted to the same volume at different doses.

[0126] The administration groups of Example 1 were intragastrically administered with the water extract of Example 1 at doses of 3 g / kg, 1.5 g / kg and 0.75 g / kg, respectively. The intragastric dose was 1 mL / 100 g, and the administration was continuous for 30 days. During the period, the blood was taken from the eye orbit every week, the whole blood was centrifuged at 3500 r / min for 10 min, the upper serum was taken, and the uric acid, creatinine and urea nitrogen contents were detected by automatic biochemical analyzer. The body weight of the rats was measured and recorded every week.

[0127] Experimental results: As can be seen from the results of Table 1 and Table 2, the model drug potassium oxonate can significantly increase the contents of blood uric acid and creatinine (P<0.001), indicating that the modeling is successful. The positive drug allopurinol can very significantly reduce the contents of blood uric acid and creatinine (P<0.001). The water extract of Stenzia radicans (high dose group, middle dose group and low dose group) of Example 1 can also very significantly reduce the contents of blood uric acid and creatinine (P<0.001). As can be seen from the results of Table 3, there is no statistical difference in the body weight of each group (P>0.05), and the water extract of Stenzia radicans of Example 1 does not cause body weight loss and other negative effects while reducing uric acid. The above shows that Stenzia radicans has a very significant uric acid-lowering effect and small side effects.

[0128] Table 1. Effect of Stenzia radicans on uric acid level of high uric acid model rats

[0129]

[0130] Table 2. Effect of Stenzia radicans on creatinine and urea nitrogen levels of high uric acid model rats

[0131] Group Dose Scr (μmol / L) BUN (mmol / L) blank group 55.16±2.15 10.90±0.53 model group 1000 mg / kg 65.38 ± 4.39 aaa ]] 10.90±0.35 positive drug group 27 mg / kg 57.25 ± 2.97 bbb ]] 10.87±0.45 low dose group 0.75 g / kg 56.21 ± 4.48 bbb ]] 10.68±0.36 middle dose group 1.5 g / kg 55.96 ± 4.36 bbb ]] 10.72±0.32 high dose group 3 g / kg 53.24 ± 4.16 bbb ]] 10.60±0.37

[0132] Table 3. Effects of P. cordata on body weight of hyperuricemia model rats

[0133] Group Dose 0 day (g) 7 day (g) 14 day (g) 21 day (g) 28 day (g) 30 day (g) blank group 214.06±8.54 277.60±12.27 329.83 18.30 358.31 23.84 383.41±33.38 397.56±30.07 model group 1000 mg / kg 220.01±4.60 274.68±6.57 325.51±9.68 353.40±13.53 385.03±15.22 390.15±15.85 positive drug group 27 mg / kg 220.21±7.34 273.00±13.77 323.99±19.90 350.73±27.47 380.38±32.44 388.22±35.04 low dose group 0.75 g / kg 219.70±5.29 278.63±10.97 335.06±21.97 362.80±25.58 394.93±32.47 401.71±31.73 middle dose group 1.5 g / kg 218.95±7.28 273.31±9.05 327.44±24.84 350.13±25.09 377.58±26.64 382.60±28.85 high dose group 3 g / kg 222.63±6.51 275.50±8.18 329.01±14.15 353.50±24.80 386.40±29.78 391.95±30.79

[0134] Data are expressed as Mean ± SE, n = 8 in each group, compared with the blank group, a P < 0.05, aa P < 0.01, aaa P < 0.001; compared with the model group, b P < 0.05, bb P < 0.01, bbb P < 0.001.

[0135] In summary, P. cordata or its extract (e.g. water extract) has the effect of lowering uric acid, and has little side effects.

[0136] Although embodiments of the present application have been disclosed as above, it is not limited only to the uses listed in the specification and embodiments, and can be fully applied to various fields suitable for the present application, and additional modifications can be easily made by those skilled in the art, and thus the present application is not limited to specific details and the examples shown and described herein, without departing from the general concept defined by the claims and the equivalent scope.

Claims

1. The use of Viola yedoensis water extract as the sole active ingredient in the preparation of drugs for lowering uric acid.

2. The use as described in claim 1, wherein the total flavonoid content of the Viola yedoensis water extract is 0.15-0.2% by weight.

3. Use of a composition comprising a hydrochloric acid extract of Viola yedoensis in the preparation of a medicament for lowering uric acid, wherein the hydrochloric acid extract of Viola yedoensis is the sole active ingredient of the composition, and the composition further comprises excipients.

4. The use as described in claim 3, wherein the total flavonoid content of the Viola yedoensis water extract is 0.15-0.2% by weight.

5. The use as described in claim 3, wherein the excipient is dextrin and / or sucralose.

6. The use as described in claim 3, wherein the composition is formulated as a decoction, tea, granules, pills, powder, tablets, capsules or oral liquid.

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