Preparation method of fructus arctii extract and application of fructus arctii extract in antipyretic and analgesic drugs
By extracting and preparing burdock extract from burdock seeds, the side effects and safety of existing antipyretic analgesic drugs were solved, significant antipyretic analgesic effects were achieved, and a high-safe drug regimen was demonstrated.
Patent Information
- Application Number
- CN202411792344.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-06
- Publication Date
- 2025-02-21
AI Technical Summary
Existing antipyretic and analgesic drugs may cause side effects during use and lack alternatives with significant safety and efficacy, especially in the treatment of fever and painful diseases.
Burdock extract was isolated and extracted from dried ripe fruits of burdock, and prepared by ultrasonic extraction of ethanol aqueous solution, under reduced pressure concentration and macroporous adsorption resin column purification, to prepare antipyretic and analgesic drugs.
Burdock extract significantly reduced anal temperature in mouse models, had significant antipyretic effects, and showed higher safety and analgesic effects by regulating inflammatory factors and reducing pain response.
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Figure CN119488540A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of traditional Chinese medicine, and in particular to the preparation of a burdock fruit extract and application thereof in antipyretic and analgesic drugs. Background Art
[0002] Fever and pain are common symptoms during the course of the disease. Fever refers to the pyrogen acting on the body's temperature regulation center, causing dysfunction of the temperature regulation center and leading to symptoms of high body temperature. Pain is a complex, unpleasant feeling that is usually accompanied by actual or potential tissue damage.
[0003] Currently, commonly used antipyretic analgesics such as aspirin, ibuprofen, acetaminophen, etc. have certain efficacy in relieving fever and pain symptoms. However, irrational use of these drugs may cause certain side effects, including gastrointestinal discomfort, liver and kidney damage, etc. In view of this, exploring antipyretic analgesics with significant efficacy and good safety has become a new research direction for scholars in related fields. Chinese herbal medicine has good development prospects due to its advantages of high efficiency, low toxicity, and low price, and has become the focus of research on new antipyretic analgesics at home and abroad.
[0004] At present, some Chinese herbal medicines or Chinese herbal compound prescriptions have been proven to have good antipyretic and analgesic effects in clinical practice, and have relatively few side effects. For example, in the treatment of colds, fever, arthritis pain and other diseases, the application of Chinese herbal medicines can reduce the dependence on non-steroidal anti-inflammatory drugs and reduce the risk of adverse drug reactions. Burdock seeds are the dried mature fruits of the Chinese herbal medicine Arctium Lappa L., which has the effects of dispersing wind-heat, clearing the lungs and relieving rashes, detoxifying and relieving throat pain. It is mainly used to treat exogenous wind-heat, cough and asthma, sore throat, carbuncle, and ulcer. Modern pharmacological studies have shown that burdock seeds have anti-inflammatory, anti-tumor, antiviral and anti-diabetic activities. However, there are few reports on the antipyretic and analgesic effects of burdock seeds. Therefore, further exploring the medicinal value of burdock seeds in antipyretic and analgesic effects can not only provide new ideas for the processing, preparation, development and utilization of burdock seeds, but also provide new solutions for the treatment of febrile and painful diseases. Summary of the invention
[0005] The purpose of the present invention is to provide a preparation method of burdock fruit extract and application of the extract in antipyretic and analgesic drugs, so as to provide a new solution for the treatment of febrile and painful diseases.
[0006] In order to achieve the above purpose, the following technical solutions are adopted:
[0007] A method for preparing a burdock fruit extract, wherein the burdock fruit extract is separated and extracted from the dried mature fruit of the Chinese medicinal herb Arctium Lappa L. of the Asteraceae family. The specific preparation method is as follows:
[0008] (1) preparing an extract: crushing and sieving burdock seeds, and ultrasonically extracting them with ethanol-water solution;
[0009] (2) preparing a concentrated solution: concentrating the above extract under reduced pressure, recovering the solvent until there is no alcohol taste, and then diluting it with 95% ethanol and purified water;
[0010] (3) Separation and purification: The concentrated solution is passed through a macroporous adsorption resin column, and purified water is used to elute and remove impurities. The ethanol water eluate is collected, concentrated and dried under reduced pressure to obtain the burdock fruit extract.
[0011] Preferably, the ethanol aqueous solution in step (1) refers to an ethanol aqueous solution with an ethanol volume fraction of 40% to 80%.
[0012] Preferably, in step (1), the weight ratio of burdock seeds to ethanol aqueous solution is 1:5-15.
[0013] Preferably, in step (1), the ultrasonic extraction temperature is 30°C to 60°C, the time is 0.5h to 1.5h, the power is 200W to 360W, and the extraction times are 1 to 3 times.
[0014] Preferably, the dilution in step (2) is to dilute with 95% ethanol and purified water to form a concentrated solution with an ethanol volume fraction of 0 to 10%.
[0015] Preferably, the macroporous adsorption resin column in step (3) is one or more of AB-8, FL-2, and D-101, and the ethanol volume fraction of the collected ethanol water eluate is 20% to 50%.
[0016] The present invention also claims to protect the use of the burdock fruit extract in the preparation of antipyretic and analgesic drugs.
[0017] The medicine uses the burdock fruit extract as the only active ingredient, or is a medicine composition containing the burdock fruit extract.
[0018] Preferably, the drug can be made into tablets, granules, dispersible tablets, injections and other preparations with conventional pharmaceutical excipients in pharmacy.
[0019] It has been verified that the burdock seed extract of the present invention has an antipyretic effect in a mouse dry yeast fever model and an analgesic effect in a mouse acetic acid pain model. The beneficial effects of the present invention are specifically shown as follows:
[0020] (1) The preparation process of burdock fruit extract is simple and quick, and the solvent and resin used are suitable for industrial production.
[0021] (2) The median lethal dose of burdock extract in mice is greater than 6g / kg, indicating that it is relatively safe.
[0022] (3) In the in vivo antipyretic experiment on mice, burdock seed extract can significantly reduce the rectal temperature of mice and has a significant antipyretic effect.
[0023] (4) In the in vivo antipyretic experiment on mice, the antipyretic effect of the burdock fruit extract prepared by macroporous resin was better than that of the simple burdock fruit ethanol-water ultrasonic extract.
[0024] (5) Burdock extract can reduce the levels of IL-6 and TNF-α in the serum of febrile mice and cAMP and PGE2 in the hypothalamus, which indicates that burdock extract can reduce inflammatory factors and regulate the body temperature center to exert an antipyretic effect.
[0025] (6) In the in vivo analgesic experiment on mice, burdock extract can significantly reduce the number of times mice twist their bodies, showing a significant analgesic effect.
[0026] (7) In an in vivo mouse analgesia experiment, the analgesic effect of burdock fruit extract prepared by macroporous resin was better than that of simple burdock fruit ethanol-water ultrasonic extract. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 To investigate the effect of burdock seed extract on body temperature in yeast-induced fever model mice.
[0028] Figure 2 Effects of different doses of burdock extract on body temperature of yeast-induced fever model mice. DETAILED DESCRIPTION
[0029] The present invention is described in detail below in conjunction with specific embodiments, but the implementation of the present invention is not limited thereto. The percentages described in the present invention are all volume fraction percentages.
[0030] Preparation Example 1 of Burdock Fruit Extract
[0031] (1) Preparation of extract: Grind burdock seeds through an 80-mesh sieve, and extract with 50% ethanol by ultrasonic. The extraction time is 30 min, the extraction temperature is 50° C., the extraction power is 300 W, the solid-liquid ratio is 1:15, and the extraction is performed once.
[0032] (2) Preparation of concentrated solution: The above extract was concentrated under reduced pressure, the solvent was recovered until there was no alcohol taste, and it was diluted with 95% ethanol and purified water to prepare a 0-10% ethanol concentrated solution containing 0.14 g / mL of crude drug.
[0033] (3) Separation and purification: The above concentrate was passed through an AB-8 macroporous adsorption resin column, impurities were removed with 4BV purified water, eluted with 5BV 50% ethanol, and the eluate was collected at a flow rate of 1-2BV / h. The burdock fruit extract was obtained by concentration and drying under reduced pressure.
[0034] Preparation Example 2 of Burdock Fruit Extract
[0035] 1) Preparation of extract: pulverize burdock seeds and pass through an 80-mesh sieve, and extract with 50% ethanol by ultrasonic method. The extraction time is 30 min, the extraction temperature is 50° C., the extraction power is 300 W, the solid-liquid ratio is 1:10, and the extraction is repeated twice.
[0036] 2) Preparation of concentrated solution: The above extract is concentrated under reduced pressure, the solvent is recovered until there is no alcohol taste, and it is diluted with 95% ethanol and purified water to prepare a 0-10% ethanol concentrated solution containing 0.14 g / mL of crude drug.
[0037] 3) Separation and purification: The above concentrate was passed through an AB-8 macroporous adsorption resin column, eluted with 4BV purified water and 4BV 20% ethanol respectively, and the ethanol eluate was collected at a flow rate of 1-2BV / h. The burdock fruit extract was obtained by concentration and drying under reduced pressure.
[0038] Preparation of ethanol aqueous solution ultrasonic extract Comparative Example 1
[0039] The burdock seeds were crushed and passed through an 80-mesh sieve, and ultrasonically extracted with 50% ethanol. The extraction time was 30 minutes, the extraction temperature was 50°C, the extraction power was 300W, and the solid-liquid ratio was 1:15. The extract was concentrated and dried under reduced pressure to obtain the product.
[0040] Experiment 1: Antipyretic effect of burdock extract on yeast-induced fever model mice
[0041] (1) Experimental Materials
[0042] Male 20-24g SPF Kunming mice (provided by Changsha Tianqin Biotechnology Co., Ltd.), main instruments: Corning electronic thermometer D-10.
[0043] (2) Experimental modeling and drug administration
[0044] The mice were adaptively raised for 3 days. The rectal temperature of the mice was measured once a day in the morning and evening 3 days before the experiment, and the mice with temperature fluctuations (ΔT>0.5°C) were eliminated. The rectal temperature was measured twice 1 hour before the experiment, and the average value was taken as the basal body temperature. The experimental animals were divided into 6 groups, 10 in each group, namely blank group, model group, aspirin group, example 1 group, example 2 group and comparative example 1 group.
[0045] The blank group was subcutaneously injected with normal saline (10mL / kg). The other groups were subcutaneously injected with 20% dry yeast suspension (10mL / kg). The rectal temperature of the mice was recorded 4, 5, 6, 7, and 8 hours after the injection of dry yeast suspension, and the temperature changes were calculated. Drug intervention was given after recording the rectal temperature at the 5th hour. Example 1 group, Example 2 group and Comparative Example 1 group were gavaged with 200mg / kg, 200mg / kg, and 400mg / kg of the corresponding drugs, respectively, the aspirin group was given 200mg / kg of aspirin, and the model group was given an equal amount of 0.5% sodium carboxymethyl cellulose.
[0046] (3) Indicator observation
[0047] The rectal temperature of mice was recorded 4, 5, 6, 7, and 8 hours after injection of dry yeast suspension.
[0048] (4) Experimental results
[0049] After injection of dry yeast suspension, the rectal temperature of mice in the model group increased significantly at 5, 6, 7, and 8 hours, indicating that the modeling was successful; compared with the model group, the rectal temperature of mice in each administration group at 6, 7, and 8 hours decreased to varying degrees, but the comparison group 1 was only statistically significant at 6 and 8 hours (P<0.05), and the rectal temperature of mice in the example group 1 and example group 2 decreased significantly at 6, 7, and 8 hours (P<0.01 or P<0.05), indicating that the burdock extract enriched and purified by macroporous resin has a better antipyretic effect than the ethanol water extract. The experimental results are as follows Figure 1 shown.
[0050] Experiment 2: Antipyretic effect of different doses of burdock extract on yeast-induced fever model mice
[0051] (1) Experimental Materials
[0052] Male 20-24g SPF Kunming mice (provided by Changsha Tianqin Biotechnology Co., Ltd.), main instruments: Corning electronic thermometer D-10.
[0053] (2) Experimental modeling and drug administration
[0054] The mice were adaptively raised for 3 days. The rectal temperature of the mice was measured once a day in the morning and evening 3 days before the experiment, and the mice with temperature fluctuations (ΔT>0.5°C) were eliminated. The rectal temperature was measured twice 1 hour before the experiment, and the average value was taken as the basal body temperature. The experimental animals were divided into 6 groups, 10 in each group, namely the blank group, the model group, the aspirin group, the low-dose group of Example 1, the medium-dose group of Example 1, and the high-dose group of Example 1.
[0055] The blank group was subcutaneously injected with normal saline (10mL / kg). The other groups were subcutaneously injected with 20% dry yeast suspension (10mL / kg). The rectal temperature of the mice was recorded 4, 5, 6, and 7 hours after the injection of the drug solution, and the temperature change compared with the basal body temperature was calculated. Drug intervention was given after recording the rectal temperature at the 5th hour. Example 1 The low, medium, and high groups were gavaged with 125mg / kg, 250mg / kg, and 500mg / kg of drugs, respectively, the aspirin group was given aspirin 200mg / kg, and the blank group and the model group were given an equal amount of 0.5% sodium carboxymethyl cellulose.
[0056] After the last temperature measurement, blood was collected, centrifuged, and serum was collected and stored at -20°C; the hypothalamus was removed, prepared into a 10% hypothalamus homogenate, and stored at -20°C.
[0057] (3) Indicator observation
[0058] (1) Record the rectal temperature of mice 4, 5, 6, and 7 hours after injection of the drug.
[0059] (2) ELISA kits were used to detect the levels of IL-6, TNF-α in mouse serum and cAMP and PGE2 in the hypothalamus.
[0060] (4) Experimental results
[0061] Compared with the blank group, the rectal temperature of mice in the model group increased significantly at 5, 6, and 7 hours, indicating that the modeling was successful; compared with the model group, the rectal temperature of mice in the low, medium, and high dose groups of Example 1 decreased significantly at 6 and 7 hours (P<0.01 or P<0.05). Figure 2 .
[0062] Compared with the blank group, the levels of IL-6, TNF-α in the serum of the mice in the model group, as well as cAMP and PGE2 in the hypothalamus, were significantly increased (P < 0.01). Compared with the model group, the levels of IL-6, TNF-α in the serum of each administration group, as well as cAMP and PGE2 in the hypothalamus, were significantly decreased (P < 0.01 or P < 0.05). The experimental results show that the extract of Example 1 exerts an antipyretic effect by regulating pyrogenic factors and central mediators of body temperature. The results are shown in Table 1.
[0063] Table 1 Effects of burdock extract on serum TNF-α, IL-6, hypothalamic cAMP, PGE2 in yeast-induced fever mice
[0064]
[0065] Compared with the blank group: # P<0.05 ## P<0.01; compared with the model group: * P<0.05 **P<0.01
[0066] Experiment 3 Analgesic effect of burdock extract on acetic acid-induced pain model mice
[0067] (1) Experimental Materials
[0068] Male 20-24g SPF grade Kunming mice (provided by Changsha Tianqin Biotechnology Co., Ltd.)
[0069] (2) Experimental modeling and drug administration
[0070] The mice were adaptively raised for 3 days. The experimental animals were divided into 5 groups, 10 in each group, namely the model group, aspirin group, example 1 group, example 2 group and comparative example 1 group. Example 1 group, example 2 group and comparative example 1 group were gavaged with 200 mg / kg, 200 mg / kg and 400 mg / kg of the drug, respectively. The aspirin group was given 200 mg / kg of aspirin, and the model group was given an equal amount of 0.5% sodium carboxymethyl cellulose. 30 minutes after administration, all mice were intraperitoneally injected with 1% glacial acetic acid (0.1 ml / 10 g), and the number of mouse twisting was observed 5-15 minutes after injection. Calculate the pain inhibition rate. Calculation formula: inhibition rate (%) = (average number of twisting times in the model group - average number of twisting times in the drug administration group) / average number of twisting times in the model group × 100%.
[0071] (3) Indicator observation
[0072] The number of body twists within 5-15 minutes after the mice were intraperitoneally injected with 1% glacial acetic acid.
[0073] (4) Experimental results
[0074] After injection of 1% glacial acetic acid, the mice in the model group showed writhing phenomenon, indicating that the modeling was successful. Compared with the model group, the number of writhing times of mice in each drug-treated group was significantly reduced, indicating that each drug-treated group had an analgesic effect. Compared with comparative example 1, the pain inhibition rate of Example 1 and Example 2 groups was higher, indicating that the analgesic effect of the burdock extract prepared by macroporous resin was better than that of the simple ultrasonic extract of burdock ethanol aqueous solution. The experimental results are shown in Table 2.
[0075] Table 2 Effects of burdock extract on acetic acid writhing pain model
[0076] Group Dosage (mg / kg) Number of twists Inhibition rate (%) Model Group - 30.83±2.14 - Example 1 Group 200 14.00±4.97 54.59 Example 2 Group 200 9.17±4.75 70.27 Comparative Example 1 400 19.00±4.98 38.38 Aspirin group 200 10.17±4.12 67.03
[0077] Experiment 4: Acute toxicity test of burdock fruit extract
[0078] (1) Experimental animals
[0079] Thirty SPF Kunming mice weighing 20-24 g (provided by Changsha Tianqin Biotechnology Co., Ltd.), half male and half female.
[0080] (2) Grouping and Observation
[0081] The mice were divided into three groups. The first group was orally administered with 6 g / kg of the extract of Example 1, the second group was orally administered with 9 g / kg of the extract of Example 1, and the third group was orally administered with the same volume of 5% dimethyl sulfoxide (prepared with 0.5% sodium carboxymethyl cellulose) as a control group. After administration, all mice were observed for two consecutive weeks, mainly observing the mice's breathing, fur color, defecation, activity, reaction to the outside world, and death. After two weeks, the mice were killed and the changes in the various organs of the mice were observed by naked eye after dissection.
[0082] (3) Test results
[0083] When the dose reached 6g / kg, the activity response of 2 mice was weakened, and the other mice were no different from the control group. After 12 hours, the mice with weakened activity returned to normal, and no mice died within 14 days; when the dose reached 9g / kg, 1 mouse died within 2 hours, and the activity response of 2 mice was weakened. The other mice were no different from the control group. After 12 hours, the mice with weakened activity returned to normal, and no mice died within 14 days. No obvious pathological changes were observed in the organs of each group of mice. The test results show that the median lethal dose of the extract of Example 1 in mice is higher than 6g / kg, indicating that the burdock extract has a high safety. The results of mouse death are shown in Table 3.
[0084] Table 3 Acute toxicity test
[0085] Group Dosage (g / kg) Number of animals Number of deaths (individuals) Example 1 6 10 0 Example 1 9 10 1 Control group - 10 0
[0086] In view of the above efficacy experiment and acute toxicity experiment results, the burdock fruit extract of the present invention has significant antipyretic and analgesic effects and good safety, and has the prospect of preparing clinical antipyretic and analgesic drugs.
Claims
1. A method for preparing a burdock fruit extract, characterized in that: The preparation method comprises the following steps: (1) preparing an extract: crushing and sieving burdock seeds, and ultrasonically extracting them with ethanol-water solution; (2) preparing a concentrated solution: concentrating the above extract under reduced pressure, recovering the solvent until there is no alcohol taste, and then diluting it with 95% ethanol and purified water; (3) Separation and purification: The concentrated solution is passed through a macroporous adsorption resin column, and purified water is used to elute and remove impurities. The ethanol water eluate is collected, concentrated and dried under reduced pressure to obtain the burdock fruit extract.
2. The method for preparing the burdock fruit extract according to claim 1, characterized in that: In the step (1), the ethanol aqueous solution refers to an ethanol aqueous solution with an ethanol volume fraction of 40% to 80%.
3. The method for preparing the burdock fruit extract according to claim 1, characterized in that: In the step (1), the weight ratio of burdock seeds to ethanol aqueous solution is 1:5-15.
4. The method for preparing the burdock fruit extract according to claim 1, characterized in that: In the step (1), the ultrasonic extraction method is: temperature 30°C to 60°C, time 0.5h to 1.5h, power 200W to 360W, and extraction times 1 to 3 times.
5. The method for preparing the burdock fruit extract according to claim 1, characterized in that: In the step (2), the dilution is performed by diluting with 95% ethanol and purified water to form a concentrated solution with an ethanol volume fraction of 0 to 10%.
6. The method for preparing the burdock fruit extract according to claim 1, characterized in that: In the step (3), the model of the macroporous adsorption resin column is one or more of AB-8, FL-2, and D-101, and the volume fraction of ethanol in the collected ethanol water eluate is 20% to 50%.
7. Use of the burdock fruit extract according to any one of claims 1 to 6 in the preparation of antipyretic and analgesic drugs.
8. The use of the burdock fruit extract in the preparation of antipyretic and analgesic drugs according to claim 7, characterized in that: The medicine uses burdock fruit extract as the only active ingredient.
9. The use of the burdock fruit extract in the preparation of antipyretic and analgesic drugs according to claim 7, characterized in that: The medicine is a medicine composition containing burdock fruit extract.
10. The use of the burdock fruit extract in the preparation of antipyretic and analgesic drugs according to claim 7, characterized in that: The medicine is prepared into tablets, granules, dispersible tablets or injections.