CN115957279B - 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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Classifications
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- 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
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a composition with xanthine oxidase inhibition effect and application thereof. The invention provides a composition with xanthine oxidase inhibition effect, which consists of mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract, and researches show that the composition obtained by reasonably proportioning the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract has remarkable inhibition effect on xanthine oxidase, and the drug effect is superior to the simple superposition of the components used independently or the combination of different components, and has remarkable synergistic effect; the four raw materials of the composition are all purely natural plant traditional Chinese medicines, the toxic and side effects are low, the harm to human bodies can be reduced, the price is relatively low, and the composition can be used as an active ingredient 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, to a composition having xanthine oxidase inhibitory effect and uses thereof.
Background
Hyperuricemia (Hyperuricemia, HUA) is a common metabolic disorder, uric acid is a nitrogenous component in urine, is a poorly soluble end product of human protein and purine metabolism, and is caused by purine metabolic dysregulation, resulting in increased uric acid production in the blood, often characterized by elevated serum uric acid levels. Uric acid is a byproduct of amino acid metabolism, and the breakdown of amino acids produces uric acid in the liver. In addition, the decomposition of purine releases small amounts of uric acid. Hyperuricemia is difficult to cure completely, long-term administration is required, and uric acid in human urine and blood can form sharp crystals, which also increases the risk of gout.
Xanthine Oxidase (XOD) is believed to be a key enzyme that causes elevated uric acid in the body, and can continuously oxidize xanthine and hypoxanthine to uric acid, thereby causing elevated uric acid in the body. If the medicine can inhibit the activity of xanthine oxidase, the medicine can achieve the effect of reducing uric acid, thereby further achieving the effect of treating hyperuricemia. Gout, hyperuricemia and other diseases are caused by accumulation in the body because uric acid which is a final product of metabolism in the human body is not decomposed, so the search for xanthine oxidase inhibitors has important significance for clinical treatment of the diseases.
However, the existing xanthine oxidase inhibitor drugs such as allopurinol, febuxostat and other chemical drugs have large toxic and side effects, possibly cause serious diseases such as liver and kidney function injury and the like, have high cost and are limited to a certain extent in application. Therefore, it is necessary to find new xanthine oxidase inhibitors. Compared with chemical medicines, the Chinese patent medicine has higher safety, stable curative effect and small toxic and side effects, and has long Chinese herbal medicine application history in China; therefore, in order to achieve the treatment or prevention of hyperuricemia and related diseases, there is a need to develop more xanthine oxidase inhibitor drugs which have less side effects and better therapeutic effects. As disclosed in the prior art, a natural composition for reducing uric acid, uses thereof and xanthine oxidase inhibitor, wherein cinnamon, licorice, guava stem and the like are adopted to inhibit xanthine oxidase, but the inhibition effect is still to be improved; the other Chinese medicinal compositions capable of inhibiting xanthine oxidase are prepared by compounding more than four different Chinese medicinal components, the medicinal properties and the medicinal effects of the Chinese medicinal components are required to be reasonably matched, the preparation method is complex, the efficacy of the compound is too wide, and the inhibition effect of the compound is also required to be improved. Therefore, the research and development of more pure natural traditional Chinese medicine compositions with stable fruits, small toxic and side effects and relatively low price and xanthine oxidase inhibition effect has important significance for inhibiting xanthine oxidase and reducing uric acid.
Disclosure of Invention
The invention aims to overcome the defects of large toxic and side effects, unstable curative effect and high cost of the existing xanthine oxidase inhibitor medicines and provide a composition prepared from pure natural plant extracts and capable of inhibiting xanthine oxidase and application thereof.
It is a first object of the present invention to provide a composition having xanthine oxidase inhibitory effect.
It is a second object of the present invention to provide the use of said composition.
It is a fourth object of the present invention to provide a xanthine oxidase inhibitor.
The above object of the present invention is achieved by the following technical scheme:
The invention provides a composition with xanthine oxidase inhibiting effect, which consists of mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract. The research shows that the composition obtained by compounding four Chinese herbal medicine extracts, namely the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract has 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, and the efficacy is obviously superior to the simple superposition of the independent use of each component or the matched use of different components, and the composition has 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 harm 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 asiatic centella herb 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 asiatic centella herb 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 2:2:2: 1.
Preferably, the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the 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 herb, centella asiatica and alpinia oxyphylla, adding an alcohol solvent for soaking, refluxing, decompressing and filtering, recovering the alcohol solvent through decompressing and distilling to obtain wet extract, respectively obtaining mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract, and mixing the four extracts according to a proportion.
Preferably, the alcohol solvent is 8-12 times of 30-65% ethanol. The 8-12 times of the amount refers to the ratio of the volume of the solvent to the mass of the raw materials, for example, 1g of the crushed traditional Chinese medicine raw materials are added with 8-12 mL of alcohol solvent.
More preferably, the crushed mulberry leaves, plantain herb, centella asiatica and alpinia oxyphylla are respectively added with 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol which are 10 times of the weight of the crushed mulberry leaves, plantain herb, centella asiatica and alpinia oxyphylla in sequence, and respectively soaked, refluxed and filtered under reduced pressure.
The invention provides application of the composition in preparation of xanthine oxidase inhibitors and uric acid reducing medicines.
The present invention provides a xanthine oxidase inhibitor comprising the above composition having a xanthine oxidase inhibitory effect.
The invention also provides application of the inhibitor in preparing medicines 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 reducing effect, the drug effect is obviously superior to the simple superposition of the single use of each component or the matched 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 injury to human bodies, are relatively low in price, can overcome the defects and defects of high toxic and side effects, unstable curative effect and high cost of the traditional xanthine oxidase inhibitor medicines, and have a wide application prospect.
Detailed Description
The invention will be further illustrated by the following specific examples in connection with the description, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
The invention is used for four plants extracted by ethanol: folium Mori, herba plantaginis, herba Centellae, fructus Alpinae Oxyphyllae are all purchased from Beijing Tongren Tang, and are washed clean and sun-dried for use.
Mulberry leaf (Folium Mori) is derived from Moraceae plant Morus alba, is a dry whole leaf, and is a medicinal plant, and contains flavone, mulberry alkaloid, polysaccharide, amino acid and microelements. The mulberry leaf has the effects of resisting tumor, resisting hypotension, resisting inflammation, resisting diabetes, reducing blood sugar and blood lipid, and regulating intestinal flora.
Plantago asiatica (Plantago asiatica L.) is also known as Daon, eichhornia crassipes, plantago asiatica, etc., and is a plant of Plantago genus of Plantaginaceae. The main components of herba plantaginis include flavonoid glycoside, phenols, phenethyl alcohol glycoside, tetraterpenes, steroid, alkaloid, polysaccharide, etc. Herba plantaginis has antiinflammatory, antibacterial, antiviral, blood sugar reducing, blood lipid reducing, and diuretic effects.
Centella asiatica (CENTELLA ASIATICA (L.) Urban) is a dried whole herb of centella asiatica belonging to the family Umbelliferae. The centella asiatica comprises tetraterpenes, volatile oils, flavonoids, sterols, alkaloids, lignin, polysaccharides, quercetin, etc. Centella asiatica has liver protecting, islet protecting, antiulcer, and anticancer effects.
Intelligence development (Alpinia oxyphylla Miq), a perennial herb of the genus alpinia of the family zingiberaceae. Has effects of warming spleen, relieving diarrhea, controlling salivation, warming kidney, reducing urination, and stopping nocturnal emission. It is commonly indicated for deficiency-cold in spleen and stomach, vomiting, diarrhea, cold pain in abdomen, excessive saliva and saliva, enuresis due to kidney deficiency, frequent urination, spermatorrhea and white turbidity.
The ethanol, the methanol, the xanthine oxidase, the dimethyl sulfoxide, the allopurinol, the sodium hydroxide, the dipotassium hydrogen phosphate, the ethylenediamine tetraacetic acid and the potassium dihydrogen phosphate adopted by the invention are all purchased from Shanghai Ala Latin Biochemical technology Co., ltd, the methanol is analytically pure, and all the 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 mulberry leaves, plantain herb and centella asiatica in a pulverizer for standby, sequentially adding 10 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol into the mulberry powder, plantain herb powder and centella asiatica powder respectively, soaking for 24 hours, 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 24 hours to obtain ethanol extract freeze-dried powder for standby.
The composition provided in this example was prepared in a mass ratio of 2:2:2: 1, respectively taking 200g of mulberry leaf extract, 200g of plantain herb extract, 200g of centella asiatica extract and 100g of alpinia oxyphylla extract, and mixing the four extracts.
Example 2 preparation of the composition
Respectively pulverizing folium Mori, herba plantaginis and herba Centellae in pulverizer, sequentially adding 8 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol into the powder, respectively, soaking for 24 hr, refluxing for 3 times, filtering under reduced pressure, distilling under reduced pressure to recover ethanol to obtain wet extract, and lyophilizing the wet extract in freeze dryer for 24 hr to obtain ethanol extract lyophilized powder.
The composition provided in this example was prepared in a mass ratio of 2:2:2: 1, respectively taking 200g of mulberry leaf extract, 200g of plantain herb extract, 200g of centella asiatica extract and 100g of alpinia oxyphylla extract, and mixing the four extracts.
Example 3 preparation of the composition
Respectively pulverizing folium Mori, herba plantaginis and herba Centellae in pulverizer, sequentially adding 12 times of 30% ethanol, 40% ethanol, 45% ethanol and 60% ethanol into the powder, respectively, soaking for 24 hr, refluxing for 3 times, filtering under reduced pressure, distilling under reduced pressure to recover ethanol to obtain wet extract, and lyophilizing the wet extract in freeze dryer for 24 hr to obtain ethanol extract lyophilized powder.
The composition provided in this example was prepared in a mass ratio of 2:2:2: 1, respectively taking 200g of mulberry leaf extract, 200g of plantain herb extract, 200g of centella asiatica extract and 100g of alpinia oxyphylla extract, and mixing the four extracts.
Example 4 preparation of the composition
In this example, a plurality of groups of compositions having different components were prepared at the same time, and the composition components are shown in Table 1 below, and the preparation methods of the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the nootropic extract are the same as in example 1.
TABLE 1 preparation of compositions of different ingredients
Example 5 inhibition Activity of compositions against xanthine oxidase
1. Solution configuration
(1) Phosphate buffer solution: 28.2mg of EDTA and 6.94g K 2HPO4、0.96g KH2PO4 mg of ultrapure water are precisely weighed, 400mL of ultrapure water is added, ultrasonic dissolution is carried out for 20min until the solution is clear and transparent, and when the pH of the solution is measured to be 7.4, the ultrapure water is used for constant volume.
(2) Xanthine oxidase solution: 2mL of 100U/mL xanthine oxidase solution was placed in a refrigerator at 6deg.C for use, and phosphate buffer solution was added to dilute to 0.10U/mL before use.
(3) Xanthine solution: 7.96mg of xanthine is weighed, added into a 50mL volumetric flask, 600 mu L of 1mol/L NaOH solution is added, ultrasonic dissolution is promoted until the solution is clear, phosphate buffer solution is used for constant volume, and ultrasonic treatment is carried out for 30min, so as to obtain xanthine solution for standby.
(4) Sample solution: the compositions prepared in examples 1 to 4 were dissolved in phosphate buffer so that the final concentrations of the sample solutions were 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
And respectively setting an enzyme reaction group, a blank group, an enzyme inhibition group and a control group, wherein the specific setting and dosage of each group are shown in table 2, setting 3 parallel measurement compound holes, detecting by using a biochemical analyzer, and taking an average value.
Table 2 test groups and dosages
The xanthine oxidase and phosphate buffer solution or ethanol extract are added into each group, and after incubating for 5min, xanthine is added to start the reaction. The test was also performed with the allopurinol positive control group.
Wherein A1, A2, A3 and A4 correspond to absorbance at 295nm of the enzyme reaction group, the blank group, the enzyme inhibition group and the control group, respectively.
3. Test results
The inhibition results of xanthine oxidase by different compositions are shown in the following table 3, wherein examples 1 to 3 are compositions obtained by different preparation methods, comparative examples 1 to 14 are compositions with different components, and allopurinol group is a positive control, and is a xanthine oxidase inhibitor commonly used at present.
TABLE 3 inhibition of xanthine oxidase by different compositions
The compositions of examples 1 to 3 and comparative examples 1 to 14 were found to have significant differences in the ability to inhibit xanthine oxidase. In examples 1 to 3, the amount of the alcohol solvent used affects the inhibition rate of the composition, and only the composition prepared by using 10 times 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 inhibition rate was small with increasing concentration. While the inhibition effect on xanthine oxidase of the composition of any combination of the two extracts of comparative examples 5 to 10 and the composition of any combination of the three extracts of comparative examples 11 to 14 was remarkably lower than that of the four extracts of examples 1 to 3, although the inhibition effect was gradually increased. The composition obtained by compounding the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract has a good xanthine oxidase inhibition effect, can reduce uric acid, has a drug effect obviously superior to simple superposition of single components or the combination of different components, and has a remarkable synergistic effect.
Example 6 optimization of compositions with xanthine oxidase inhibition
The mass ratio of the composition was optimized in this example, and the preparation method was the same as in 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, and the specific arrangement is shown in the following table 4, and the method for measuring xanthine oxidase inhibition was the same as in example 5.
TABLE 4 mass ratios of the different extracts
The inhibition effect of compositions with different component ratios on xanthine oxidase is measured at the same time, and the result is shown in table 5, and the inhibition effect on xanthine oxidase can be changed by adjusting the mass ratio of each extract in the composition on the premise that the dosage of each composition is the same. The mass ratio of mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract adopted in the composition is 2:2:2:1, the xanthine oxidase inhibitory effect was best, closest to the positive control allopurinol group.
TABLE 5 inhibition of xanthine oxidase by compositions of different component ratios
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (6)
1. A composition with xanthine oxidase inhibiting effect is characterized by comprising mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract; the mass ratio of the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract is (1-3): (1-3): (1-3): (1-3);
The mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the alpinia oxyphylla extract are all alcohol extracts;
The preparation method of the composition comprises the following steps: respectively taking mulberry leaves, plantain herb, centella asiatica and alpinia oxyphylla, adding an alcohol solvent for soaking, refluxing, decompressing and filtering, recovering the alcohol solvent through decompressing and distilling to obtain wet extract, and then freeze-drying to respectively obtain mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract, and mixing the mulberry leaf extract, plantain herb extract, centella asiatica extract and alpinia oxyphylla extract according to a proportion to obtain the composition with xanthine oxidase inhibition effect; the alcohol solvent adopts 30-65% ethanol with the amount of 8-12 times.
2. The composition according to claim 1, wherein the mass ratio of the mulberry leaf extract, the plantain herb extract, the centella asiatica extract and the nootropic extract is (2 to 3): (2-3): (2-3): (1-2).
3. Use of a composition according to claim 1 or 2 for the preparation of a xanthine oxidase inhibitor.
4. Use of a composition according to claim 1 or 2 for the manufacture of a uric acid lowering medicament.
5. A xanthine oxidase inhibitor, characterized in that it is prepared from a composition according to claim 1 or 2.
6. Use of the inhibitor according to claim 5 for the preparation of a medicament for the treatment of hyperuricemia or gout.
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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 |
Non-Patent Citations (2)
| Title |
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| 积雪草水提液对高尿酸血症模型大鼠尿酸代谢的影响;胡向阳;刘桃丽;林春淑;杨璇;;中医杂志;第58卷(第01期);第60-63页 * |
| 积雪车前口服液对痛风模型鸡的疗效研究;翟福展等;动物医学进展;第40卷(第4期);第135页左栏第2段、第138页倒数第1段 * |
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