Traditional Chinese medicine composition with effect of treating gout as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention discloses a traditional Chinese medicine composition for treating gout as well as a preparation method and application of the traditional Chinese medicine composition. The raw material medicines comprise the following components in parts by weight: 53-70 parts of morinda citrifolia fruits, 17-35 parts of haws and 13-19 parts of citrus chachiensis hortorum. The pharmaceutical composition disclosed by the invention has a repairing effect on the growth of EA.hy926 cells inhibited by sodium urate, has analgesic and anti-inflammatory effects on gouty arthritis model rats, reduces the level of uric acid in serum, is simple in medicine and special in effect, and has a wide application prospect.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a Chinese medicinal composition having the effect of treating gout, and a preparation method and application thereof. Background Art

[0002] Gout is a chronic disease caused by the accumulation of urate crystals in joints and surrounding tissues. Its typical manifestations are recurrent arthritis and the potential disability that follows. Since 1990, the global prevalence of gout has shown a significant upward trend. Between 1990 and 2019, the global prevalence of gout increased by 63.44%, while gout-related disability-adjusted life years (DALYs) also increased by 51.12%. Analysis based on an age-period-cohort model shows that the prevalence of gout increases steadily with age and gradually increases in younger birth cohorts. The global prevalence of gout is expected to continue to rise in the future.

[0003] Traditional Chinese medicine not only has rich clinical experience in the prevention and treatment of gout, but also has outstanding efficacy. It can effectively relieve and eliminate patients' clinical symptoms, improve their quality of life, and has significant advantages in preventing recurrence. For example, Chinese invention patent application 118526553A discloses a Chinese medicinal composition for treating gout with the effects of removing dampness and turbidity, removing blood stasis and dredging collaterals. The composition of the Chinese medicinal composition is as follows: 8-12 parts of Smilax glabra, 5-9 parts of Polygonum cuspidatum, 5-9 parts of Plantago seed, 5-9 parts of stir-fried Atractylodes macrocephala, 5-9 parts of Phellodendron amurense, 1-4 parts of Cyathula root, 1-4 parts of Coix seed, 1-4 parts of Coix lacryma-jobi, 1-4 parts of Codonopsis pilosula, 1-4 parts of Fructus Smilacis Glabrae, 1-4 parts of Cortex qinpi, 1-4 parts of Hedyotis diffusa, 1-4 parts of Rhubarb, 1-4 parts of Caulis Achyranthis Bidentatae, 1-4 parts of Sargentodoxae, 1-4 parts of Caulis Trachelospermi, 1-4 parts of Lonicera japonica, 1-4 parts of Carthamus tinctorius, 1-4 parts of stir-fried peach kernel, 1-4 parts of Lithospermum officinale, 1-4 parts of Salvia miltiorrhiza, 1-4 parts of Stephania tetrandra, and 18-12 parts of roasted Astragalus membranaceus; Chinese invention patent application 118161578A discloses a traditional Chinese medicine combination with the effects of removing dampness, relieving numbness, calming wind and relieving pain, as well as its preparation method and use, wherein the traditional Chinese medicine combination comprises: 95-135 parts of Radix Catnip, 95-125 parts of Trillium, 95-135 parts of Fraxinus officinalis, 95-135 parts of Gentiana macrophylla, 95-135 parts of Plantago seed, 145-210 parts of Poria, 40-70 parts of Rhizoma Alismatis, 310-380 parts of Semen Coicis, 40-70 parts of Beehive, 95-135 parts of Rhizoma Dioscoreae, 125-165 parts of Herba Siegesbeckiae, and 40-70 parts of Eupolyphaga sinensis. However, the therapeutic effects of the drugs in the above patent documents on gout need to be further improved.

[0004] Therefore, there is an urgent need to develop a Chinese medicine composition for treating gout that is highly safe, simple in medicine, and more effective. Summary of the Invention

[0005] Gout is a disease that falls under the category of arthritis. Its causes involve many aspects, such as insufficient human vital energy, imbalance of qi and blood, obstruction of meridians, internal accumulation of phlegm and blood stasis, and stagnation in joints and tendons. In view of the existing diagnosis and treatment system for gout, the present invention provides a Chinese medicine composition for treating gout composed of Noni fruit, Crataegus pinnatifida, and Noni fruit. In the Chinese medicine composition of the present invention, dried tangerine peel can eliminate dampness and resolve phlegm, and can be used for all symptoms of qi stagnation, dampness obstruction, and phlegm congestion, and can clear the phlegm and dampness congestion in the meridians and joints. Guangdong dried tangerine peel is an authentic Chinese medicinal material from the production area (Xinhui, Guangdong) among the Chinese medicinal dried tangerine peels, and is the best quality among the dried tangerine peels. Crataegus pinnatifida can promote qi and invigorate the spleen, disperse blood stasis and resolve turbidity, and can help dried tangerine peel clear the body of blood stasis and turbid phlegm, and also regulate the spleen and promote qi and blood circulation, harmonizing qi and blood. Noni fruit can clear away heat and relieve pain to treat the symptoms, and nourish yin and promote body fluid to treat the root cause. All the medicines are used together to promote qi and invigorate the spleen and nourish yin, remove dampness and dredge the meridians and relieve pain. According to the results of pharmacological experiments and in vitro experiments, the Chinese medicine composition of the present invention has anti-inflammatory and analgesic effects, is highly safe, simple in medicine and effective, and can be used to prevent and treat gout.

[0006] The technical solution of the present invention to solve the above problems is as follows:

[0007] A traditional Chinese medicine composition with the effect of treating gout, the composition is composed of traditional Chinese medicine extracts and medically acceptable excipients. The raw materials used are all common commercial products, and their sources are not specifically limited. The traditional Chinese medicine composition comprises the following components by weight: 53-70 parts of noni fruit, 17-35 parts of hawthorn, and 13-19 parts of dried tangerine peel.

[0008] In the above scheme, the optimal ratio of the raw materials is: 53 parts of noni fruit, 28 parts of hawthorn, and 19 parts of dried tangerine peel.

[0009] A second object of the present invention is to provide a method for preparing a Chinese medicine composition having the effect of treating gout, comprising the following steps:

[0010] The raw materials are washed and decocted with water, the decoction is filtered to remove the drug residue, concentrated, refluxed with ethanol, centrifuged, and lyophilized. The lyophilized powder is dissolved in water and dialyzed with a dialysis bag. The dialyzate of polysaccharide with a molecular weight greater than 5000 Daltons is collected, concentrated, and lyophilized to obtain the pharmaceutical composition.

[0011] Preferably, the decoction is to soak each raw material drug in an appropriate amount of clean water for 30-60 minutes, then add water and decoct twice, the first time adding 11 times the water and decocting for 1 hour; the second time adding 10 times the water and decocting for 1 hour, filtering through gauze or filter paper to remove the drug residue, combining the filtrate to obtain a clarified medicinal solution.

[0012] Preferably, the dialysis is performed using a 5000 Dalton dialysis bag with flowing water.

[0013] The present invention also provides the use of the above-mentioned traditional Chinese medicine composition in preparing medicine for treating gout.

[0014] The Chinese medicine composition of the present invention can be a common oral preparation, such as tablets, capsules, granules, pills, syrups or oral emulsions.

[0015] The pharmaceutical composition of the present invention has a repairing effect on the growth of EA.hy926 cells inhibited by sodium urate, and has analgesic and anti-inflammatory effects on gouty arthritis model rats, reduces the level of uric acid in serum, is simple in medicine and has a specific effect, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Response surface diagram and contour diagram of the interaction effect of three factors on cell survival rate. DETAILED DESCRIPTION

[0017] The present invention is described in detail below with reference to specific embodiments.

[0018] Noni fruit: The fruit of Morinda citrifolia L. Take the original medicinal material, remove impurities, and dry it.

[0019] Hawthorn: The dried mature fruit of Crataegus pin.nati fida Bge., a plant of the Rosaceae family. The fruit is harvested in autumn when ripe, removed of impurities and fallen pits, washed, and dried.

[0020] Guang Chenpi (Guang Chenpi): Dried mature peel of Citrus reticulata 'Chachi' (Rutaceae). Remove impurities from the original herb, spray with water, soak thoroughly, cut into strips, and dry in the shade.

[0021] Example 1

[0022] This embodiment provides a Box-Behnken response surface optimization experiment for the pharmaceutical composition with the effect of treating gout. The three factors of noni fruit addition, hawthorn addition, and tangerine peel addition were selected as independent variables, and a three-factor three-level experiment was designed using Box-Behnken. The experimental factors and levels are detailed in Table 1. The raw materials were taken according to the addition amount in Table 1, washed, soaked in an appropriate amount of clean water for 30-60 minutes, and then decocted twice with water. The first time, 11 times the water was added and decocted for 1 hour; the second time, 10 times the water was added and decocted for 1 hour. The filtrates were combined and filtered through gauze or filter paper to remove the residue to obtain a clear medicinal solution. The extract was collected and concentrated into a concentrated solution; ethanol was added to the obtained concentrated solution, refluxed, centrifuged, and precipitated and freeze-dried; the obtained freeze-dried powder was dissolved in water and dialyzed with a 5000 Dalton dialysis bag with flowing water. The polysaccharide dialysate with a molecular weight greater than 5000 Dalton was collected, concentrated, and freeze-dried to obtain the polysaccharide extract. After sodium urate inhibited EA.hy926 cell growth (cell viability was 72.34 ± 2.45%) using the CCK-8 assay (Chen Weijia. Screening and Mechanism of Action of Traditional Chinese Medicine for Gout Treatment [D]. Jilin University, 2017), we evaluated whether the herbal extract could restore the inhibitory effect. The drug dosage was optimized using the EA.hy926 cell viability (%) as the response value. The response surface design experiment and results are shown in Table 2.

[0023] Table 1 Response surface experimental factors and levels

[0024]

[0025]

[0026] Table 2 Response surface design experiment and cell survival results

[0027]

[0028]

[0029] The variance analysis of the cell survival rate results was performed using Design-Expert software, and the quadratic polynomial regression equation model of A (noni fruit addition amount), B (hawthorn addition amount), and C (tangerine peel addition amount) was obtained as follows:

[0030] Cell viability = 101.1-1.18A+0.56B-0.054C-0.57AB+0.31AC+0.54BC-15.06A 2 -2.74B 2 -2.13C 2 The variance analysis of the regression model is shown in Table 3.

[0031] Table 3 Analysis of variance of regression model

[0032]

[0033]

[0034] As shown in Table 3, the regression equation model P < 0.0001 is extremely significant, the lack of fit term is not significant (P = 0.7376 > 0.05), and the model R 2 =0.9914, indicating a good fit for the model and a high correlation between the actual and predicted values. Therefore, the model can well reflect the influence of each factor on the response value and predict the optimal addition ratio. The P values ​​indicate that each quadratic term is extremely significant (P < 0.01), with the noni fruit addition level A being significant (P < 0.05), while the others are not significant. The F values ​​indicate that the order of influence of the three main factors is: A > B > C, i.e., noni fruit addition level > hawthorn addition level > tangerine peel addition level.

[0035] The quadratic polynomial regression equation given by Design-Expert software was used to draw a response surface diagram of the interaction between various factors on cell survival rate ( Figure 1 Color changes in the contour plot represent changes in the response value. Faster color changes and denser contour lines indicate a steeper slope, indicating a more significant impact on cell survival. The shape of the contour lines also reflects the strength of interactions between factors; an elliptical shape indicates significant interactions between the studied factors. Response surface equations were used to calculate the optimal drug formulation for cell survival. However, considering actual operational conditions, the optimal ratio was: 53 parts noni fruit, 28 parts hawthorn, and 19 parts dried tangerine peel.

[0036] Example 2

[0037] The invention discloses a preparation method of the present invention comprising the following steps: taking 53 parts of noni fruit, 28 parts of hawthorn, and 19 parts of dried tangerine peel, washing them, soaking them in 5 times of clean water for 45 minutes, and then decocting them twice with water: adding water to 11 times of the water for the first time and decocting them for 1 hour; adding 10 times of water for the second time and decocting them for 1 hour; combining the filtrates, filtering them through gauze or filter paper, removing the medicinal residues, and obtaining a clarified medicinal solution. The extract was collected and placed in a constant temperature oven at 60°C for 12 hours to be concentrated into a concentrate with a drug concentration of 1 g / ml; ethanol was added to the obtained concentrate until the ethanol concentration was about 75%, the mixed liquid was heated to reflux temperature (85°C), and kept under reflux for 10 hours, the mixture was naturally cooled and then centrifuged, and the precipitate was freeze-dried; the obtained freeze-dried powder was dissolved in water, dialyzed with flowing water using a 5000 Dalton dialysis bag, and the polysaccharide dialysate with a molecular weight greater than 5000 Dalton was collected, transferred to a rotary evaporator, concentrated under reduced pressure, and rotary evaporated at 55°C and 60 r / min to remove excess water, and the remaining liquid was collected, and the remaining liquid was frozen in a -80°C refrigerator, and then placed in a freeze dryer and vacuum freeze-dried for 48 hours to obtain the polysaccharide extract.

[0038] Example 3

[0039] By mass, 40 parts of Noni fruit, 35 parts of Hawthorn, and 25 parts of Guang Chenpi are taken, washed, soaked in an appropriate amount of clean water for 30-60 minutes, and then decocted twice with water. The first time, 11 times of water is added and the decocted for 1 hour; the second time, 10 times of water is added and the decocted for 1 hour. The filtrate is combined and filtered through gauze or filter paper to remove the medicinal residue to obtain a clarified medicinal liquid. The extract is collected and placed in a 60°C constant temperature oven for 12 hours to be concentrated into a drug concentration of 1g / ml concentrated solution; ethanol is added to the obtained concentrated solution to an ethanol concentration of about 75%, the mixed liquid is heated to reflux temperature (85°C), and kept under reflux for 10 hours, and the mixture is naturally cooled and then centrifuged and the precipitate is freeze-dried; the obtained freeze-dried powder is dissolved in water, dialyzed with a 5000 Dalton dialysis bag with flowing water, and the polysaccharide dialyzate with a molecular weight greater than 5000 Dalton is collected, concentrated, and freeze-dried to obtain the polysaccharide extract.

[0040] Example 4

[0041] By mass, 70 parts of Noni fruit, 17 parts of Hawthorn, and 13 parts of Guang Chenpi are taken, washed, soaked in an appropriate amount of clean water for 30-60 minutes, and then decocted twice with water. The first time, 11 times of water is added and the decocted for 1 hour; the second time, 10 times of water is added and the decocted for 1 hour. The filtrate is combined and filtered through gauze or filter paper to remove the medicinal residue to obtain a clarified medicinal liquid. The extract is collected and placed in a 60°C constant temperature oven for 12 hours to be concentrated into a drug concentration of 1g / ml concentrated solution; ethanol is added to the obtained concentrated solution to an ethanol concentration of about 75%, the mixed liquid is heated to reflux temperature (85°C), and kept under reflux for 10 hours. After the mixture is naturally cooled, it is centrifuged and the precipitation is freeze-dried; the obtained freeze-dried powder is dissolved in water, dialyzed with a 5000 Dalton dialysis bag with flowing water, and the polysaccharide dialyzate with a molecular weight greater than 5000 Dalton is collected, concentrated, and freeze-dried to obtain the polysaccharide extract.

[0042] Example 5

[0043] 500 grams of Noni fruit is taken, washed, and soaked in an appropriate amount of clean water for 30-60 minutes, then decocted twice with water, the first time with 11 times the amount of water and decocted for 1 hour; the second time with 10 times the amount of water and decocted for 1 hour, the filtrates are combined, filtered through gauze or filter paper, and the medicinal residues are removed to obtain a clarified medicinal solution. The extract is collected and placed in a 60°C constant temperature oven for 12 hours to be concentrated into a concentrated solution with a drug concentration of 1 g / ml; ethanol is added to the obtained concentrated solution to an ethanol concentration of about 75%, the mixed liquid is heated to reflux temperature (85°C), and maintained at reflux for 10 hours, and the mixture is naturally cooled and then centrifuged and the precipitate is freeze-dried; the obtained freeze-dried powder is dissolved in water, dialyzed with flowing water using a 5000 Dalton dialysis bag, and the polysaccharide dialyzate with a molecular weight greater than 5000 Dalton is collected, concentrated, and freeze-dried to obtain the polysaccharide extract.

[0044] Example 6

[0045] Take 500 grams of hawthorn, wash it, soak it in an appropriate amount of clean water for 30-60 minutes, then add water and decoct it twice, adding 11 times the amount of water for the first time and decoct it for 1 hour; add 10 times the amount of water for the second time and decoct it for 1 hour, combine the filtrates, filter through gauze or filter paper, remove the medicinal residue, and obtain a clear medicinal solution. Collect the extract and place it in a constant temperature oven at 60°C for 12 hours to concentrate it into a concentrated solution with a drug concentration of 1g / ml; add ethanol to the obtained concentrated solution until the ethanol concentration is about 75%, heat the mixed liquid to reflux temperature (85°C), and maintain reflux for 10 hours. After the mixture is naturally cooled, centrifuge it, and precipitate it and freeze-dry it; dissolve the obtained freeze-dried powder in water, dialyze it with flowing water using a 5000 Dalton dialysis bag, collect the polysaccharide dialysate with a molecular weight greater than 5000 Dalton, concentrate it, and freeze-dry it to obtain the polysaccharide extract.

[0046] Example 7

[0047] Get 500 grams of Pericarpium Citri Reticulatae, after cleaning, put into appropriate amount of clear water and soak 30-60 minute after adding water and decocting twice, add 11 times of water for the first time, decoct 1 hour; Add 10 times of water for the second time, decoct 1 hour, merging filtrate, by gauze or filter paper filtering, remove medicinal residues, obtain clarifying medicinal liquid. Collect extracting solution and place 60 ℃ of constant temperature baking ovens and be condensed into drug concentration 1g / ml concentrated solution in 12 hours; In gained concentrated solution, add ethanol to ethanol concentration approximately 75%, mixing liquid is heated to reflux temperature (85 ℃), and kept refluxing 10 hours, treat that mixture naturally cools after centrifugal, precipitation freeze-drying; Gained lyophilized powder is dissolved in water, uses 5000 daltonian dialysis tubing flowing water dialysis, collects molecular weight greater than 5000 daltonian polysaccharide dialyzate, concentrated, freeze-drying, promptly obtain described polysaccharide extract.

[0048] Example 8 (Efficacy Experiment)

[0049] Experimental Animals and Modeling: Male 8-week-old Sprague-Dawley rats weighing 300 ± 20 g were selected. A hyperuricemia model was established by oral gavage of adenine (100 mg / kg) and ethambutol hydrochloride (250 mg / kg) for 21 days. One hour after the last oral gavage, the rats in each group were anesthetized and inserted into the right ankle joint cavity with a 6-gauge needle with the bevel facing forward and upward at a 45° angle to the tibia. Except for the normal group, which was injected with an equal volume of normal saline, all other groups were injected with 0.2 mL of 0.5 g / mL monosodium urate (MSU) solution.

[0050] Animal Dosing: 80 rats with successfully established models were selected and divided into a model group, a colchicine group, and Example 2-7 groups, with 10 rats in each group. Ten healthy SD rats were also selected as a blank control group. Colchicine served as a positive control at a dose of 0.27 mg / kg. The blank control group was gavage-administered with the same volume of distilled water. The Example 2-7 group was gavage-administered with a dose of 0.475 g / kg. Each group was gavage-administered for two weeks.

[0051] After the final dose, rats were subjected to a weight-bearing differential test to assess bipedal pressure differential and infrared thermal allodynia measurement to measure leg-lifting time (the duration of leg-lifting time reflects the severity of the condition; shorter leg-lifting times indicate more severe disease, while shorter leg-lifting times indicate milder disease). The next day, rats in each group underwent autopsy, requiring 24 hours of food and water deprivation. Rats were anesthetized with an intraperitoneal injection of 10% chloral hydrate (prepared with saline) and anesthetized. After abdominal dissection, 3-5 ml of blood was collected from the abdominal aorta. The blood was then allowed to stand at 4°C overnight (for at least 12 hours). After separation, the supernatant was aspirated and serum UA and TNF-α levels were measured by enzyme-linked immunosorbent assay. The results are shown in Tables 4-5.

[0052] Table 4 Effect of the Chinese medicine composition of the present invention on the difference between the two feet of rats and the time of raising the feet

[0053] Group Difference between two feet (g) Leg lifting time (S) Blank control group <![CDATA[4.24±1.51 # *]]> <![CDATA[18.367±1.128* # ]]> Model Group <![CDATA[63.61±5.61 # ]]> <![CDATA[6.13±0.87 # ]]> Colchicine group 32.81±3.87* <![CDATA[15.05±0.69 # ]]> Example 2 group 33.72±4.87* 15.55±1.28* Example 3 group <![CDATA[42.58±2.73 # *]]> <![CDATA[12.25±1.29 # *]]> Example 4 Group <![CDATA[44.34±2.16 # *]]> <![CDATA[11.55±1.78 # *]]> Example 5 group <![CDATA[48.92±2.85 # *]]> <![CDATA[11.81±0.95 # *]]> Example 6 <![CDATA[50.62±3.92 # *]]> <![CDATA[10.59±1.08 # *]]> Example 7 Group <![CDATA[51.51±1.86 # *]]> <![CDATA[10.12±1.12 # *]]>

[0054] Compared with the model group: *P<0.05, significant difference; compared with Example 2 group: # P<0.05, significant difference.

[0055] As shown in Table 4, after gout modeling, the difference between the two feet and the time of lifting the feet of the rats increased significantly (P<0.05), and the groups of Examples 2 to 7 were able to significantly improve the difference between the two feet and the time of lifting the feet of the rats, and the effect of the group of Example 2 was significantly better than that of the other groups of Examples, and the difference was significant (P<0.05).

[0056] Table 5 Effect of the Chinese medicine composition of the present invention on the levels of TNF-α and UA in rat serum

[0057]

[0058]

[0059] Compared with the model group: *P<0.05, significant difference; compared with Example 2 group: # P<0.05, significant difference.

[0060] As shown in Table 5, after gout modeling, the TNF-α and UA levels in the rat serum increased significantly (P<0.05), while the Examples 2 to 7 groups were able to significantly reduce the TNF-α and UA levels in the rat serum, and the effect of the Example 2 group was better than that of the other Example groups, and the difference was significant (P<0.05).

Claims

1. A Chinese medicine composition for treating gout, characterized in that: The raw material medicine comprises the following components by weight: 53-70 parts of noni fruit, 17-35 parts of hawthorn, and 13-19 parts of tangerine peel.

2. The Chinese medicine composition according to claim 1, characterized in that: The raw materials are: 53 parts of noni fruit, 28 parts of hawthorn, and 19 parts of dried orange peel.

3. A method for preparing the Chinese medicine composition for treating gout according to claim 1, characterized in that: The following steps are involved: The raw materials are washed and decocted with water, the decoction is filtered to remove the drug residue, concentrated and refluxed with ethanol, centrifuged, precipitated and freeze-dried, the freeze-dried powder is dissolved in water, dialyzed with a dialysis bag, the polysaccharide dialysate with a molecular weight greater than 5000 Daltons is collected, concentrated and freeze-dried to obtain the pharmaceutical composition.

4. The preparation method according to claim 3, characterized in that: The decoction is to soak each raw material medicine in appropriate amount of clean water for 30-60 minutes, then add water and decoct twice, add 11 times of water for the first time and decoct for 1 hour; add 10 times of water for the second time and decoct for 1 hour, filter through gauze or filter paper, remove the drug residue, combine the filtrate, and obtain a clear drug solution.

5. The preparation method according to claim 3, characterized in that: The dialysis is performed by using a 5000 Dalton dialysis bag with flowing water.

6. Use of the Chinese medicine composition according to claim 1 in preparing medicine for treating gout.

7. The use according to claim 6, characterized in that: The Chinese medicine composition is in the form of tablets, capsules, granules, pills, syrups or oral emulsions.

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