CN115957279A - Composition with xanthine oxidase inhibition effect and application thereof - Google Patents
Composition with xanthine oxidase inhibition effect and application thereof Download PDFInfo
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- CN115957279A CN115957279A CN202211538895.5A CN202211538895A CN115957279A CN 115957279 A CN115957279 A CN 115957279A CN 202211538895 A CN202211538895 A CN 202211538895A CN 115957279 A CN115957279 A CN 115957279A
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- xanthine oxidase
- centella asiatica
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a composition with xanthine oxidase inhibition effect and application thereof. Researches show that the composition obtained by reasonably proportioning the mulberry leaf extract, the plantain herb extract, the asiatic centella herb extract and the alpinia oxyphylla extract has obvious inhibition effect on the xanthine oxidase, and the drug effect is superior to that of simple superposition of single use of each component or matched use of different components, so that the composition has obvious synergistic effect; the four raw materials of the composition are all pure natural plant traditional Chinese medicines, have low toxic and side effects, can reduce the harm to human bodies, have relatively low price, and can be used as active ingredients for preparing xanthine oxidase inhibitors and uric acid reducing medicines.
Description
Technical Field
The present invention belongs to the field of medicine, health product and food technology. More particularly, relates to a composition with xanthine oxidase inhibiting effect and application thereof.
Background
Hyperuricemia (HUA) is a common metabolic disease, uric acid is a nitrogen-containing component in urine, is a poorly soluble end product of human protein and purine metabolism, is caused by purine metabolic disorder, and causes an increase in uric acid production in blood, and is generally characterized by an increase in serum uric acid levels. Uric acid is a by-product of the metabolism of amino acids, the breakdown of which produces uric acid in the liver. In addition, the breakdown of purines releases small amounts of uric acid. Hyperuricemia is difficult to completely cure, long-term administration is required, and uric acid in human urine and blood can form sharp crystals and also increase the risk of gout.
Xanthine Oxidase (XOD) is considered to be a key enzyme causing an increase in uric acid in the body, and it can continuously oxidize xanthine and hypoxanthine to convert into uric acid, thereby causing an increase in uric acid in the body. If the medicament can inhibit the activity of xanthine oxidase, the medicament can achieve the effect of reducing uric acid, and further has the effect of treating hyperuricemia. The diseases such as gout, hyperuricemia and the like are caused by that the final product uric acid metabolized in a human body is not decomposed and accumulated in the human body, so that the search for the xanthine oxidase inhibitor has important significance for the clinical treatment of the diseases.
However, the current xanthine oxidase inhibitors such as allopurinol and febuxostat have serious toxic and side effects, which may cause serious diseases such as liver and kidney function damage, and the cost is high, which is limited in application. Therefore, the search for new xanthine oxidase inhibitors is necessary. Compared with chemical medicines, the Chinese patent medicine has higher safety, stable curative effect and small toxic and side effect, and China has a long history of Chinese herbal medicine application; therefore, in order to treat or prevent hyperuricemia and related diseases, it is highly desirable to develop a xanthine oxidase inhibitor which has fewer side effects and a better therapeutic effect. For example, the prior art discloses a natural composition for reducing uric acid, application thereof and a xanthine oxidase inhibitor, cinnamon, liquorice, guava stem and the like are adopted to inhibit xanthine oxidase, but the inhibition effect is still to be improved; the other traditional Chinese medicine compositions capable of inhibiting xanthine oxidase are obtained by compounding more than four different traditional Chinese medicine components, the medicine properties and the medicine effects of the traditional Chinese medicine components need to be reasonably matched, the preparation method is complex, the efficacy in the compound formed by the traditional Chinese medicine compositions is too wide, and the inhibition effect is to be improved. Therefore, the research and development of more pure natural traditional Chinese medicine compositions with stable effect, less toxic and side effect and relatively low price and the xanthine oxidase inhibition effect have important significance for inhibiting the xanthine oxidase and reducing the uric acid.
Disclosure of Invention
The invention aims to overcome the defects and shortcomings of large toxic and side effects, unstable curative effect and high cost of the existing xanthine oxidase inhibitor drugs, and provides a composition capable of inhibiting xanthine oxidase and application thereof, wherein the composition is prepared from pure natural plant extracts.
The first object of the present invention is to provide a composition having xanthine oxidase inhibitory action.
A second object of the invention is to provide the use of said composition.
The fourth purpose of the invention is to provide a xanthine oxidase inhibitor.
The above purpose of the invention is realized by the following technical scheme:
the invention provides a composition with xanthine oxidase inhibiting effect, which consists of mulberry leaf extract, plantain herb extract, asiatic centella extract and alpinia oxyphylla extract. The research shows that the composition obtained by compounding the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the fructus alpiniae oxyphyllae extract has a remarkable xanthine oxidase inhibition effect, so that the conversion of xanthine and hypoxanthine into uric acid is reduced, the uric acid reducing effect is good, the drug effect is obviously superior to that of the simple superposition of the single use of each component or the matching use of different components, and the composition has a remarkable synergistic effect. The composition provided by the invention is extracted from four pure natural plant traditional Chinese medicines, has low toxic and side effects, can reduce the damage to human bodies, is easy to obtain and has low price.
Preferably, the mass ratio of the mulberry leaf extract to the plantain herb extract to the centella asiatica extract to the alpinia oxyphylla extract is (1-3) to (1-3): (1-3).
More preferably, the mass ratio of the mulberry leaf extract to the plantain herb extract to the centella asiatica extract to the alpinia oxyphylla extract is (2-3) to (2-3): (1-2).
Further preferably, the mass ratio of the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract is 2:1.
preferably, the mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract are all alcohol extracts.
The invention provides a preparation method of the composition, which comprises the steps of respectively taking mulberry leaves, plantain, centella asiatica and fructus alpiniae oxyphyllae, crushing, adding an alcohol solvent for soaking, refluxing, filtering under reduced pressure, recovering the alcohol solvent through reduced pressure distillation to obtain wet extracts, freeze-drying to respectively obtain mulberry leaf extracts, plantain extracts, asiatic centella extracts and fructus alpiniae oxyphyllae extracts, and mixing the four extracts according to a proportion to obtain the composition.
Preferably, the alcohol solvent adopts 8-12 times of 30-65% ethanol. The amount of the solvent which is 8 to 12 times of the total weight of the traditional Chinese medicine is the ratio of the volume of the solvent to the mass of the raw materials, for example, 8 to 12mL of alcohol solvent is added into 1g of the crushed traditional Chinese medicine raw materials.
More preferably, the crushed mulberry leaves, plantain, centella asiatica and fructus alpiniae oxyphyllae are respectively added into 10 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol for soaking, refluxing and filtering under reduced pressure.
The invention provides application of the composition in preparation of a xanthine oxidase inhibitor and preparation of a medicament for reducing uric acid.
The invention provides a xanthine oxidase inhibitor, which contains the composition with xanthine oxidase inhibition effect.
The invention also provides application of the inhibitor in preparation of a medicine for treating hyperuricemia or gout.
The invention has the following beneficial effects:
the research of the invention shows that the composition obtained by compounding the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract has good xanthine oxidase inhibition effect and good uric acid reduction effect, the drug effect is obviously superior to the simple superposition of the single use of each component or the matching use of different components, the inhibition rate can reach 88.94 percent, and the composition has obvious synergistic effect. The four extracts in the composition are all extracted from pure natural plant traditional Chinese medicines, have low toxic and side effects, can reduce the damage to human bodies, have relatively low prices, can overcome the defects and the defects of large toxic and side effects, unstable curative effect and high cost of the existing xanthine oxidase inhibitor medicines, and have wide application prospects.
Detailed Description
The present invention is further illustrated by the following specific examples in the specification, which are not intended to limit the invention in any way. The reagents, methods and apparatus employed in the present invention are conventional in the art, except as otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
The present invention is used for four plants for ethanol extraction: mulberry leaves, plantain herbs and centella asiatica are purchased from Beijing Tongrentang, washed clean and dried in the sun for later use.
Folium Mori (Folium Mori) is dry whole leaf of Morus alba L of Moraceae, is a medicinal plant, and contains flavone, folium Mori alkaloid, polysaccharide, amino acids and microelements. Folium Mori has anti-tumor, hypotension resisting, antiinflammatory, antidiabetic, blood sugar lowering, blood lipid reducing, and intestinal flora regulating effects.
Plantago asiatica L, also known as swertia, plantago asiatica, etc., is a plant of the Plantago genus of the Plantaginaceae family. The main components of herba plantaginis include flavonoid glycoside, phenols, phenethanol glycosides, tetraterpenes, steroids, alkaloids, and polysaccharide. The herba plantaginis has antiinflammatory, antibacterial, antiviral, blood sugar lowering, blood lipid reducing, and diuretic effects.
Centella Asiatica (l.) Urban) is a dried whole plant of Centella Asiatica of the family umbelliferae. The main components of centella include tetraterpenoids, volatile oils, flavonoids, sterols, alkaloids, lignins, polysaccharides, quercetin, etc. Herba Centellae has effects of protecting liver, protecting pancreas islet, resisting ulcer, and resisting cancer.
Alpinia oxyphylla Miq, a perennial herb of the genus Alpinia of the family Zingiberaceae. Has the effects of warming spleen, stopping diarrhea, controlling saliva, warming kidney, reducing urination and stopping emission. It is commonly used for deficiency-cold of spleen and stomach, vomiting, diarrhea, cold pain in abdomen, excessive saliva, enuresis due to kidney deficiency, frequent micturition, spermatorrhea and whitish and turbid urine.
The ethanol, methanol, xanthine oxidase, dimethyl sulfoxide, allopurinol, sodium hydroxide, dipotassium hydrogen phosphate, ethylene diamine tetraacetic acid and potassium dihydrogen phosphate adopted by the invention are all purchased from Shanghai Aladdin Biotechnology Co., ltd, the methanol is analytically pure, and all water used for the test is ultrapure water
Example 1 preparation of the composition
Respectively taking mulberry leaves, plantain herb and centella asiatica, pulverizing the fructus alpiniae oxyphyllae in a pulverizer for later use, then taking mulberry powder, plantain herb powder, centella asiatica powder and fructus alpiniae oxyphyllae powder, sequentially adding 10 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol into the fructus alpiniae oxyphyllae powder respectively, soaking for 24h, refluxing for 3 times, filtering under reduced pressure, distilling under reduced pressure to recover ethanol to obtain wet extract, and drying the wet extract in a freeze dryer for 24h to obtain ethanol extract freeze-dried powder for later use.
The composition provided in this example was mixed in a mass ratio of 2: respectively taking 200g of mulberry leaf extract, 200g of plantain herb extract, 200g of asiatic centella herb extract and 100g of alpinia oxyphylla extract obtained in the above steps, and mixing the four extracts to obtain the traditional Chinese medicine composition.
Example 2 preparation of the composition
Respectively taking mulberry leaves, plantain herb and centella asiatica, crushing the mulberry leaves, the plantain herb and the centella asiatica in a crusher for later use, then respectively adding 8 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol into mulberry powder, the plantain herb powder, the centella asiatica powder and the intelligence-improving powder in sequence, soaking the mixture for 24 hours, refluxing the mixture for 3 times, filtering the mixture under reduced pressure, distilling the mixture under reduced pressure to recover ethanol to obtain wet extract, and drying the wet extract in a freeze dryer for 24 hours to obtain ethanol extract freeze-dried powder for later use.
The composition provided in this example was mixed in a mass ratio of 2: respectively taking 200g of mulberry leaf extract, 200g of plantain herb extract, 200g of asiatic centella herb extract and 100g of alpinia oxyphylla extract obtained in the above steps, and mixing the four extracts to obtain the traditional Chinese medicine composition.
Example 3 preparation of the composition
Respectively taking mulberry leaves, plantain herb and centella asiatica, crushing the mulberry leaves, the plantain herb and the centella asiatica in a crusher for later use, then respectively adding 12 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol into mulberry powder, the plantain herb powder, the centella asiatica powder and the intelligence-improving powder in sequence, soaking the mixture for 24 hours, refluxing the mixture for 3 times, filtering the mixture under reduced pressure, distilling the mixture under reduced pressure to recover ethanol to obtain wet extract, and drying the wet extract in a freeze dryer for 24 hours to obtain ethanol extract freeze-dried powder for later use.
The composition provided in this example was mixed in a mass ratio of 2: respectively taking 200g of mulberry leaf extract, 200g of plantain herb extract, 200g of asiatic centella herb extract and 100g of alpinia oxyphylla extract obtained in the above steps, and mixing the four extracts to obtain the traditional Chinese medicine composition.
Example 4 preparation of the composition
This example simultaneously prepared multiple sets of different compositions, the composition ingredients of which are shown in table 1 below, and the preparation of mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract was the same as in example 1.
TABLE 1 preparation of compositions of the different ingredients
Example 5 inhibitory Activity of the composition on xanthine oxidase
1. Solution preparation
(1) Phosphate buffer solution: 28.2mg EDTA and 6.94g K are precisely weighed 2 HPO 4 、0.96g KH 2 PO 4 400mL of ultrapure water is added, ultrasonic dissolution is performed for 20min until the solution is clear and transparent, and when the pH of the solution is measured to be =7.4, the volume is determined by using the ultrapure water.
(2) Xanthine oxidase solution: a2 mL 100U/mL xanthine oxidase solution was kept in a refrigerator at 6 ℃ and diluted to 0.10U/mL with a phosphate buffer solution before use.
(3) Xanthine solution: weighing 7.96mg xanthine, adding into a 50mL volumetric flask, adding 600 mu L of 1mol/L NaOH solution, performing ultrasonic dissolution promotion until the solution is clear, performing constant volume by using phosphate buffer solution, and performing ultrasonic treatment for 30min to obtain xanthine solution for later use.
(4) Sample solution: the compositions prepared in examples 1 to 4 were each dissolved in phosphate buffered saline to give sample solutions having final concentrations of 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 400. Mu.g/mL, 600. Mu.g/mL, 800. Mu.g/mL, and 1000. Mu.g/mL, respectively, for use.
2. Test method
An enzyme reaction group, a blank group, an enzyme inhibition group and a control group are respectively arranged, specific setting and dosage of each group are shown in a table 2, 3 parallel determination multiple holes are arranged, a biochemical analyzer is used for detection, and an average value is obtained.
Table 2 test groups and dosages
The group is first added with xanthine oxidase and phosphate buffer solution or ethanol extract, incubated for 5min, and then added with xanthine to start reaction. The test was also set with an allopurinol positive control group.
In the formula, A1, A2, A3 and A4 correspond to the absorbance at 295nm of the enzyme reaction group, blank group, enzyme inhibition group and control group, respectively.
3. Test results
The results of the inhibition of xanthine oxidase by different compositions are shown in table 3 below, wherein examples 1 to 3 are compositions obtained by different preparation methods, comparative examples 1 to 14 are compositions of different components, and allopurinol group is a positive control, and is a currently commonly used xanthine oxidase inhibitor.
TABLE 3 inhibition of xanthine oxidase by different compositions
The determination shows that the compositions of examples 1 to 3 and comparative examples 1 to 14 have significant difference in the inhibition ability to xanthine oxidase. In examples 1 to 3, the amount of the alcohol solvent influences the inhibition rate of the composition, and only the composition prepared by using 10 times of the amount of the alcohol solvent in example 1 has the best inhibition effect. In comparative examples 1 to 4, the composition containing only one component had little inhibitory effect on xanthine oxidase at low concentrations (≦ 200. Mu.g/mL), and the increase in the inhibitory rate was small as the concentration increased. The composition of the two extracts of comparative examples 5 to 10 in any combination and the composition of the three extracts of comparative examples 11 to 14 in any combination have a significantly lower xanthine oxidase inhibitory effect than those of examples 1 to 3 in any combination. The composition obtained by compounding the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the fructus alpiniae oxyphyllae extract has a good xanthine oxidase inhibition effect, can reduce uric acid, has a drug effect obviously superior to that of simple superposition of single use of each component or matching use of different components, and has a remarkable synergistic effect.
Example 6 optimization of composition having xanthine oxidase inhibitory action
In this example, the mass ratio of the composition was optimized, the preparation method was the same as that of example 1, except that the mass ratio of the prepared mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract was different, the specific setup is shown in table 4 below, and the method for measuring xanthine oxidase inhibition was the same as that of example 5.
TABLE 4 Mass ratios of the different extracts
The effect of the compositions with different component ratios on the inhibition of xanthine oxidase was simultaneously determined, and the results are shown in table 5, wherein the inhibition effect on xanthine oxidase was changed by adjusting the mass ratio of each extract in the compositions under the premise of the same dosage of each composition. The mass ratio of the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract adopted in the composition is 2:2:2:1, the inhibition effect on xanthine oxidase is the best, and the inhibition effect is closest to a positive control allopurinol group.
TABLE 5 xanthine oxidase inhibition results of compositions with different component ratios
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A composition having xanthine oxidase inhibitory effect is characterized by comprising mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract.
2. The composition of claim 1, wherein the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract are mixed in a mass ratio of (1-3) to (1-3): (1-3).
3. The composition of claim 2, wherein the mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract are present in a mass ratio of (2-3) to (2-3): (1-2).
4. The composition of any one of claims 1-3, wherein the mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract are all alcohol extracts.
5. The method for preparing the composition according to any one of claims 1 to 4, wherein the composition is prepared by pulverizing mulberry leaves, plantain, centella asiatica and alpinia oxyphylla respectively, adding an alcohol solvent for soaking, refluxing, filtering under reduced pressure, recovering the alcohol solvent by distillation under reduced pressure to obtain a wet extract, freeze-drying to obtain mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract respectively, and mixing the four extracts in proportion.
6. The method as claimed in claim 5, wherein the alcohol solvent is 30-65% ethanol in an amount of 8-12 times that of the alcohol solvent.
7. Use of a composition according to any one of claims 1 to 4 for the preparation of a xanthine oxidase inhibitor.
8. Use of the composition according to any one of claims 1 to 4 for the preparation of a medicament for lowering uric acid.
9. A xanthine oxidase inhibitor comprising the composition according to any one of claims 1 to 4.
10. Use of the inhibitor of claim 9 for the preparation of a medicament for the treatment of hyperuricemia or gout.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102908403A (en) * | 2012-11-13 | 2013-02-06 | 广东新大枫化工科技有限公司 | Chinese materia medica preparation for treating gout and preparation method thereof |
| CN108882744A (en) * | 2016-04-08 | 2018-11-23 | 韩国韩医学研究院 | Contain intelligence development extract as effective component for preventing, improving or treating the composition of hyperuricemia or dysbolism relevant to hyperuricemia |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102908403A (en) * | 2012-11-13 | 2013-02-06 | 广东新大枫化工科技有限公司 | Chinese materia medica preparation for treating gout and preparation method thereof |
| CN108882744A (en) * | 2016-04-08 | 2018-11-23 | 韩国韩医学研究院 | Contain intelligence development extract as effective component for preventing, improving or treating the composition of hyperuricemia or dysbolism relevant to hyperuricemia |
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