Compositae plant extract as well as preparation method and application thereof
By extracting and purifying chrysanthemum lactone compounds from the chrysanthemum extract, the adverse reactions of existing PDE4 inhibitors in the treatment of PDE4-type-related diseases were solved, and effective relief of psoriasis and low toxicity and high safety treatment effects were achieved.
Patent Information
- Application Number
- CN202510347351.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-24
- Publication Date
- 2025-06-27
AI Technical Summary
When treating PDE4-type-related diseases, existing PDE4 inhibitors have adverse reactions such as diarrhea, nausea, etc., which limits their use and is difficult to distinguish targets of different subtypes, resulting in side effects.
Acetyl lactone compounds were extracted from the erectile dysfunction extract, and purified by alcohol extraction, extraction and column chromatography to prepare a compound with specific PDE4 inhibitory activity.
This compound significantly relieves the symptoms of psoriasis, can inhibit the expression of a variety of inflammatory factors and cytokines, reduces inflammatory skin infiltration, and has low toxicity and fewer side effects.
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Figure CN120204276A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the application field of natural extracts, and particularly relates to a preparation method and application of wedelolactone compounds. Background Art
[0002] Under certain conditions, after being damaged by etiological factors, the body will have abnormal vital activities due to the disorder of homeostasis regulation, leading to the occurrence of diseases. The treatment of diseases mainly relies on drug treatment. However, while drugs are treating diseases, they can also cause toxic and side effects, such as toxic reactions, metabolic effects, allergic reactions, etc. One of the main reasons for the occurrence of toxic and side effects of drugs is the binding of drugs to non-target targets in the body. For example, propranolol, an older antihypertensive drug, belongs to non-selective β-blockers. Since β-receptors are distributed throughout the human body and can be subdivided into β1 and β2 receptors, propranolol has an antagonistic effect on both types of receptors. The blockade of β1 receptors in the heart has the effects of lowering blood pressure and relieving arrhythmia, etc. However, the blockade of β2 receptors in the respiratory tract can cause bronchial smooth muscle contraction, resulting in reactions such as dyspnea. At the same time, the antagonism of β2 receptors in the gastrointestinal tract can cause adverse reactions such as diarrhea. These adverse reactions are caused by the binding to non-target proteins.
[0003] Phosphodiesterase type 4 (PDE4) specifically hydrolyzes cAMP to 5'-AMP, regulates the cAMP concentration in cells within an appropriate range, and maintains normal physiological activities. PDE4 is mainly distributed in various inflammatory cells, immune cells and airway smooth muscle cells in the human body. Currently, the treatment of diseases related to PDE4 type (such as atopic dermatitis, psoriasis, asthma, chronic obstructive pulmonary disease and rheumatoid arthritis, etc.) mainly aims to inhibit inflammatory cells by inhibiting the activity of PDE4 to achieve the purpose of treating diseases.
[0004] The PDE4 family can be further divided into four subtypes: PDE-4A, PDE-4B, PDE-4C, and PDE-4D. Currently, PDE4 inhibitors have been developed into anti-inflammatory drugs. However, PDE4 inhibitors generally cannot distinguish between subtypes of PDE4 and can bind to different subtypes in vivo. Among these four subtypes, PDE-4B is the main anti-inflammatory target, while PDE-4D is associated with gastrointestinal discomfort and is the main subtype that causes vomiting. When a PDE4 inhibitor binds to the anti-inflammatory target of PDE-4B in vivo, it will also bind to the target of PDE-4D, resulting in gastrointestinal discomfort and triggering adverse reactions. For example, the PDE4 inhibitor roflumilast is mainly used clinically to treat lung inflammation, especially for the treatment of asthma and chronic obstructive pulmonary disease. However, taking this type of drug can cause gastrointestinal adverse reactions such as diarrhea and nausea, limiting its use. Therefore, how to overcome the adverse reactions of these PDE4 inhibitors and study specific inhibitors has become one of the research hotspots.
[0005] Psoriasis, commonly known as "psoriasis vulgaris", is a chronic relapsing inflammatory skin disease with a polygenic genetic background and is one of the PDE4-related diseases. According to statistics, the global incidence of psoriasis is about 2%, and there are currently about 6.5 million psoriasis patients in China. The occurrence of psoriasis is accompanied by a systemic inflammatory cascade reaction in local skin lesions. In addition to affecting the skin, some psoriasis patients also suffer from metabolic diseases such as arthritis, diabetes, and hypertension.
[0006] Natural products are one of the important sources for finding specific inhibitors and are of great significance for developing a new generation of specific PDE4 inhibitors with strong efficacy and low side effects. Eclipta prostrata is the dried aerial part of the plant Eclipta prostrata L. of the Compositae family, also known as Eclipta prostrata and Jinlingcao, and has traditional effects such as nourishing the liver and kidneys, stopping bleeding and cooling blood. It belongs to the category of cooling blood and hemostatic drugs under the classification of hemostatic drugs. Modern pharmacological experimental studies have shown that Eclipta prostrata has the effects of inhibiting bacterial growth, protecting the liver, enhancing the body's immunity, anti-osteoporosis, lowering blood sugar, and lowering blood lipids. Currently, the research on it mainly focuses on chemical components, pharmacological effects, in vivo metabolism, quality control, active ingredients, and other related preparation processes and pharmacological activities. After retrieval, no relevant literature has been found on using the active ingredients of Eclipta prostrata as specific PDE4 inhibitors. Summary of the Invention
[0007] Aiming at the technical defects existing in the above-mentioned prior art, the purpose of the present invention is to provide wedelolactone compounds in the extract of Eclipta prostrata, which have strong efficacy, low toxicity and side effects, and have a significant effect of relieving psoriasis symptoms.
[0008] The above object of the present invention is achieved by the following technical solutions:
[0009] In the first aspect of the present invention, there is provided an application of an extract of Compositae plants in the treatment and / or prevention of diseases related to phosphodiesterase type 4, and the extract of Compositae plants is an extract of Eclipta prostrata L..
[0010] Further, the extract of Eclipta prostrata L. is obtained by alcohol extraction of Eclipta prostrata L., and the method of alcohol extraction is as follows: Weigh dry leaves of Eclipta prostrata L., crush them into coarse powder, soak them three times with the same volume of alcohol solution, wherein the first time is 2 - 5 days, and the second and third times are each 1 - 3 days, filter, combine the filtrates, recover the effective components of alcohol under reduced pressure, extract the above recovered product with petroleum ether and ethyl acetate, recover the effective components under reduced pressure, and concentrate the remaining liquid to a thick paste to obtain an alcohol extract of Eclipta prostrata L. containing the effective part.
[0011] Further, there is provided an application of wedelolactone compounds in the extract of Compositae plants in the treatment and / or prevention of diseases related to phosphodiesterase type 4.
[0012] Further, the wedelolactone compounds are one of compound 1 and compound 2 shown in the following formula I, the structural formula of compound 1 is 1 in the following formula I, and the structural formula of compound 2 is 2 in the following formula I;
[0013]
[0014] In the second aspect of the present invention, there is provided a preparation method of wedelolactone compounds, including the following steps:
[0015] A. Crush dry leaves of Eclipta prostrata L., soak them three times with methanol or ethanol solution, filter, combine and collect the filtrates, and then concentrate under reduced pressure to a paste to obtain an alcohol extract of Eclipta prostrata L..
[0016] B. Make the alcohol extract into a suspension with distilled water, extract it successively with petroleum ether and ethyl acetate, and concentrate the ethyl acetate extraction solution under reduced pressure to obtain an ethyl acetate extraction paste.
[0017] C. Subject the ethyl acetate extraction paste to column chromatography separation and purification to obtain monomeric compound 1 and monomeric compound 2.
[0018] Further, the step C includes:
[0019] 1) Subject the ethyl acetate extraction paste to silica gel column chromatography separation, elute successively with methanol - water gradient at volume ratios of 5:95, 30:70, 55:45, 80:20, and 95:5, and collect the methanol - water eluates at volume ratios of 30:70 and 55:45.
[0020] 2) Take the methanol-water eluates with volume ratios of 30:70 and 55:45 for ODS column chromatography, and elute with methanol-water gradient of volume ratios 20:80, 30:70, 55:45, 65:35 and 95:5. Concentrate the methanol-water eluates with volume ratios of 30:70 and 55:45 respectively.
[0021] 3) Take the methanol-water eluate for purification by semi-preparative HPLC, using methanol-water as the mobile phase with volume ratio (95 - 5):(5 - 95) to obtain Compound 1 and Compound 2.
[0022] The third aspect of the present invention is to provide an application in the preparation of health products or foods for treating and / or preventing psoriasis, wherein the health products or foods contain the wedelolactone compounds.
[0023] The fourth aspect of the present invention is to provide a drug for treating and / or preventing psoriasis, wherein the drug contains one or both of the Compositae plant extract or the wedelolactone compounds with any of the shown structures, as well as pharmaceutically acceptable excipients and carriers.
[0024] Furthermore, the dosage form of the drug is capsule, tablet, pill, granule, infusion, spray, emulsion and ointment.
[0025] The present invention has the following beneficial effects:
[0026] 1. The wedelolactone compounds extracted from the extract of Eclipta prostrata L. of the present invention have a significant effect of alleviating psoriasis symptoms, and can simultaneously inhibit the expression of a variety of inflammatory factors, cytokines and genes related to cutin envelope formation, so as to relieve hyperkeratosis with parakeratosis, erythema and scales of psoriasis, etc., and reduce symptoms such as inflammatory infiltration of the skin. Therefore, it has good development and application prospects in the preparation of psoriasis treatment drugs.
[0027] 2. The raw materials are derived from natural plants, with little irritation to the skin lesions, low toxicity and few side effects. Brief Description of the Drawings
[0028] Figure 1 It is the NMR data diagram of monomer Compound 1 in the embodiment of the present invention.
[0029] Figure 2 It is the NMR data diagram of monomer Compound 2 in the embodiment of the present invention.
[0030] Figure 3 It is the treatment effect diagram of monomer Compound 2 on psoriasis mice in the embodiment of the present invention.
[0031] Figure 4 It is the representative image of HE staining and quantification of the stratum corneum of mice in the embodiment of the present invention.
[0032] Figure 5 The acute toxicity effect of monomer compound 2 of the embodiment of the present invention on the digestive system.
[0033] Figure 6 The content change diagram of aspartate aminotransferase (AST), uric acid (UA), and blood urea nitrogen (UREA) in mice of the embodiment of the present invention.
[0034] Figure 7 The viscerosomatic ratio diagram of the heart, liver, spleen, lung, and kidney of the dissected mice in the embodiment of the present invention.
[0035] Figure 8 The skin section diagram of mice in the embodiment of the present invention. Specific embodiments
[0036] The present invention will be further described below in conjunction with embodiments, but the embodiments do not limit the present invention in any form. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in the technical field.
[0037] In the following embodiments, the NMR spectrum was recorded using a Bruker 400 MHz nuclear magnetic resonance spectrometer with TMS as the internal standard. Mitsubishi Chemical MCI packing (CHP20P, 75 - 150 μM), YMC ODS packing (12 nm, S - 50 μM); the rest of the solvents and reagents: analytical pure (AR).
[0038] Example 1 Extraction, separation process of Eclipta prostrata and its inhibitory activity against PDE4
[0039] Weigh the dry leaves of Eclipta prostrata, crush them into coarse powder, and soak them three times with the same volume of alcohol solution. Among them, the first time is 2 - 5 days, and the second and third times are each 1 - 3 days. Filter, combine the filtrates, recover the effective components of alcohol under reduced pressure, and concentrate the remaining solution to a thick paste to obtain the methanol extract of Eclipta prostrata containing the effective part.
[0040] The methanol extract of Eclipta prostrata was suspended in 4 times the mass of water, and successively extracted with petroleum ether and ethyl acetate with the same volume as water (each solvent was continuously extracted 3 times) to obtain petroleum ether, ethyl acetate extracts and aqueous solution. They were respectively evaporated to dryness under reduced pressure to obtain petroleum ether extract, ethyl acetate extract and aqueous concentrate, and then the inhibitory activities against PDE4 of different components were tested respectively.
[0041] The PDE4 enzyme activity was measured by liquid scintillation counting method. As a cAMP - preferred isozyme, PDE4 was selected 3The inhibitory activity of PDE4 was tested using H-cAMP as a substrate. The test method is outlined as follows: Prepare Assay buffer (50 mM Tris-HCl, 10 mM MgCl2 / MnCl2, 1.0 mM DTT, pH 7.5); Dilute the PDE4D protein (recombinant expressed by the research group, for the specific method, see the published literature: Biochemical Pharmacology 130(2017)51–59) with Assay buffer to an appropriate multiple and place it on ice for later use; Dilute the substrate 3 H-cAMP to an appropriate concentration (the reading of the control group is about 20000 cpm); Dissolve and dilute the test compound with DMSO to the set concentration; Take 58 μL of the diluted substrate into a 1.5 mL Eppendorf tube, add 2 μL of the prepared test compound at the set concentration, mix well, then add 40 μL of the diluted enzyme solution, mix gently, and then let it stand at room temperature for 15 min; Add 200 μL of 0.2 M ZnSO4 (to terminate the reaction) and 0.2 N Ba(OH)2 solution (to precipitate the ring-opened 3 H-cAMP) to the reaction system in sequence, centrifuge at high speed (14800 rpm) for 5 min, take 430 μL of the supernatant containing unreacted 3 H-cAMP and add it to 2 mL of scintillation fluid, shake well and then measure the radioactive reading with a liquid scintillation counter. Each group of experiments is set with a substrate group (C0), a target protein group (C E ), a positive control group (rolipram, C P ), and an inhibitor group (C I ). The inhibition rate of the inhibitor is calculated by the formula Inhibition Rate = [1 - (C0 - C I ) / (C0 - C E )] * 100%. To determine the specific IC 50 value (the IC 50 value is the inhibitor concentration when the inhibition rate reaches 50%), set 8 - 10 concentration gradients of sesquialter dilution according to the results of the initial measurement of the inhibition rate, measure the inhibition rate respectively, and then use GraphPad Prism software for non-linear fitting to obtain its specific IC 50 value, and repeat independently three times to obtain the mean and standard deviation. The results of the inhibitory activity of the extract of Eclipta prostrata L. on PDE4D are shown in Table 1 below.
[0042] Table 1 Inhibitory activity of the extract of Eclipta prostrata L. on PDE4D
[0043] Eclipta prostrata extract 10 μg / mL 1 μg / mL <![CDATA[IC 50 (μmol / L)]]> Total methanol extraction 95.2 49.5 n.s.b Petroleum ether 81.7 33.5 n.s.b Ethyl acetate 106.2 56.6 n.s.b Water 42.6 n.s.b n.s.b
[0044] Isolation and Identification of Wedelolactone Compounds in the Extract of Eclipta prostrata L. and Their Inhibitory Activity against PDE4
[0045] I. Preparation of Wedelolactone Monomer Compound 1 and Monomer Compound 2
[0046] Select the ethyl acetate extract with the best inhibitory activity against PDE4. First, fractionate it preliminarily with an MCI column, eluting with methanol:water = 5:95 → 95:5. After TLC detection, combine it into 2 fractions (Fr.A - Fr.B). Then, repeatedly purify it with an ODS column. Finally, further purify it by high-performance liquid chromatography under the condition of (methanol:water) to obtain 2 wedelolactone monomer compounds.
[0047] The preparation methods of monomer compound 1 and monomer compound 2 specifically include the following steps:
[0048] Dissolve 3 g of the concentrated ethyl acetate extract of Eclipta prostrata L. in 300 ml of methanol, evaporate off small-molecule polar solvents under reduced pressure, and repeat this step. Dissolve the obtained sample in 300 ml of methanol, add three times the mass of C18 silica gel powder, mix well, and evaporate under reduced pressure to obtain dry silica gel powder. First, fractionate it preliminarily with an MCI column, and perform methanol-water gradient elution at volume ratios of 5:95, 30:70, 55:45, 80:20, and 95:5 (5 ml / min, 500 ml) respectively. Detect at a wavelength of 254 nm to preliminarily purify the two compounds. When the volume ratio is 30:70, obtain fraction Fr.A. Keep the solvent gradient. After the peak is completely eluted, increase the solvent gradient. When the volume is 55:45, obtain fraction Fr.B. Keep the solvent gradient. After the peak is completely eluted, collect the solvent and evaporate under reduced pressure to obtain dry powder.
[0049] Purify fractions Fr.A and Fr.B with an ODS column, eluting with methanol-water gradient at volume ratios of 20:80, 30:70, 55:45, 65:35, and 95:5 respectively. Concentrate the methanol-water eluates with volume ratios of 30:70 and 55:45 respectively. Purify the methanol-water eluates with preparative HPLC. Use methanol-water as the mobile phase and perform gradient elution at volume ratios of 5:95, 30:70, 55:45, 80:20, and 95:5 (5 ml / min, 500 ml). Detect at a wavelength of 254 nm to obtain monomer compound 1 and monomer compound 2.
[0050] Identification of Monomer Compound 1 and Monomer Compound 2
[0051] Perform structure identification on the obtained monomer compound 1 and monomer compound 2. The results are respectively as Figure 1 and Figure 2 shown. From Figure 1 and Figure 2It can be seen that monomeric compound 1 is norwedelolactone monomeric compound, and monomeric compound 2 is wedelolactone monomeric compound. Their structures are shown as follows, where 1 represents the structure of monomeric compound 1 and 2 represents the structure of monomeric compound 2.
[0052]
[0053] II. Test on the inhibitory activity of wedelolactone monomeric compound 2 against PDE4
[0054] After obtaining the above two wedelolactone monomeric compounds, compound 2 was selected to test its inhibitory activity against PDE4 by the same method (the activity of PDE4 enzyme was measured by liquid scintillation counting method). As shown in Table 2 below.
[0055] Table 2 Inhibitory activity of wedelolactone monomeric compound 2 against PDE4D
[0056]
[0057] Example 3 Verification of the in vivo anti - psoriasis effect of the extract of Eclipta prostrata L. and wedelolactone monomeric compound 2 in the extract
[0058] The extract of Eclipta prostrata L. with inhibitory effect on PDE4 and monomeric compound 2 isolated from the extract were selected as test samples to evaluate their therapeutic effects in a psoriasis animal model. The specific experimental materials, methods and results are described as follows.
[0059] (I) Experimental materials
[0060] SPF - grade BALB / c mice, male, 6 - 8 weeks old, weighing 18 - 22 g, were purchased from Beijing Spearf Bio - technology Co., Ltd. (License number: SCXK(Beijing)2019 - 010). Temperature: 20 - 25 °C, humidity 50 - 70%, with free access to food and water. Imiquimod (Hubei Keyi Pharmaceutical Co., Ltd., 5%) and Calcipotriol (LEO Laboratories Limited).
[0061] (II) Experimental methods
[0062] BALB / c mice were randomly divided into 4 groups with 10 mice in each group. Group A - normal control group (Control), Group B - imiquimod-induced model group (IMQ, 62.5 mg applied to the back), Group C - imiquimod + positive drug calcipotriene (IMQ + Calcipotriene), Group D - imiquimod + 10% ethyl acetate extract of Eclipta prostrata (IMQ + 10% E.prostrata-EA), Group E - imiquimod + 2% monomeric compound 2 (IMQ + 2% Wedelolactone (WDL)), Group F - imiquimod + 5% monomeric compound 2 (IMQ + 5% Wedelolactone (WDL)). The back of the mice was selected as the experimental observation area. One day before the experiment, the hair on the back surface of the mice was shaved off to expose a hairless area of 2 cm × 3 cm. The control group was applied with 62.5 mg of vaseline every day, and the mice in the other groups were given 62.5 mg of 10% imiquimod ointment every day. After the drug was absorbed, the treatment drug was applied to the back of the mice in the form of an ointment. By observing the skin lesions of the mice every day, taking pictures and recording, scoring was carried out according to the PASI criterion (graded from 0 - 4) to evaluate the therapeutic effect of monomeric compound 2 on psoriasis mice.
[0063] (III) Experimental results
[0064] As Figure 3 shown in Figures A - F in Figure 3 It can be seen from Figure A in Figure 3 that after seven days of drug administration, on the seventh day, the mice showed psoriasis-like symptoms such as erythema and scales after imiquimod was applied to the back, indicating successful modeling; topical application of 10% ethyl acetate extract of Eclipta prostrata (10% E.prostrata-) ointment, 2% monomeric compound 2 (2% WDL) ointment or 5% monomeric compound 2 (5% WDL) ointment could significantly improve psoriasis symptoms;
[0065] From Figure 3As can be seen from Figures A - F, the group administered with 5% monomeric compound 2 (IMQ + 5% WDL) showed the best effect in promoting the recovery of scaling, redness, and thickening after modeling, being superior to the calcipotriene treatment group (IMQ + Calcipotriene), the ethyl acetate extract of Eclipta prostrata treatment group (IMQ + 10% E.prostrata - EA), and the group administered with 2% monomeric compound 2 (IMQ + 2% WDL), and having a relatively small impact on body weight.
[0066] As Figure 4 shown in Figures A - D. Figure 4 Figure A in it is the immunohistochemical result, Figure 4 Figure B in it is the HE staining of the mouse stratum corneum, Figure C is the expression level of CD3+, and Figure D is the expression level of TNF - a.
[0067] Figure 4 Figure A in it shows that the group administered with 5% monomeric compound 2 (IMQ + 5% WDL) can effectively increase the expression level of cAMP and effectively reduce the contents of pro - inflammatory cytokines such as IL - 17A, IL - 6, IL - 23, and TNF - a, thereby alleviating the psoriasis state in mice.
[0068] Figure 4 Figure B in it shows that the imiquimod - induced model group (IMQ) can induce epidermal hyperplasia, parakeratosis, and inflammatory cell infiltration. The application of 10% ethyl acetate extract of Eclipta prostrata (E.prostrata - EA) ointment and monomeric compound 2 (2% / 5% WDL) ointment can reduce the severity of these skin lesions.
[0069] Figure 4 In Figures C and D, the darker the color, the more severe the inflammation, the thicker the stratum corneum, and the more severe the lesion degree. From Figure 4 Figures C and D, it can be seen that the epidermal thickness of the mice in the model group (IMQ) increased significantly due to abnormal keratinocyte hyperplasia, while the application of 10% ethyl acetate extract of Eclipta prostrata (E.prostrata - EA) ointment or monomeric compound 2 (2% / 5% WDL) cream can inhibit this abnormal hyperplasia and restore the epidermal thickness to normal. Among them, in the administered groups, the 5% WDL cream had the most significant normalization effect. It indicates that WDL can effectively reduce the epidermal inflammatory response at a higher concentration.
[0070] The above - mentioned Figure 3 and Figure 4 results show that the group administered with 5% monomeric compound 2 (IMQ + 5% WDL) can effectively reduce the inflammatory state of the skin, and the effect is better than that of the positive drug group (IMQ + Calcipotriene), the EA phase group (IMQ + 10% E.prostrata - EA), and the group administered with 2% compound 2 (IMQ + 2% WDL).
[0071] Subacute Toxicity Experiment of the Cream Prepared from Monomeric Compound 2 in Mice
[0072] The monomeric compound 2 of wedelolactone was selected as the test sample to evaluate its short-term toxicity effect in normal animals. The specific experimental materials, methods and results are described as follows.
[0073] (I) Experimental Materials
[0074] SPF-grade BALB / c mice, male and female, 6 weeks old, weighing 18 - 22 g, temperature: 20 - 25 °C, humidity 50 - 70%, free access to food and water.
[0075] (II) Experimental Methods
[0076] Male BALB / c mice were randomly divided into 2 groups, namely the blank control group and the administration group containing 15% monomeric compound 2 (WDL), with 5 mice in each group; female BALB / c mice were randomly divided into 2 groups, namely the blank control group and the administration group containing 15% monomeric compound 2, with 5 and 6 mice respectively. After the quarantine period, the mice were weighed, and the back of the mice was selected as the experimental observation area. One day before the experiment, the hair on the back surface of the mice was cut off to expose a hairless area of 2 cm × 3 cm. The male blank control group and the female blank control group were smeared with blank cream once a day, and the male administration group and the female administration group were smeared with the cream containing 15% monomeric compound 2 once a day. The skin condition was observed, and the body weight, water intake and food intake were recorded. After 14 days of administration, the mice were dissected, and the blood biochemistry, visceral index, HE staining and other data results were analyzed to evaluate its short-term toxicity effect in normal animals.
[0077] (III) Experimental Results
[0078] The body weight and food intake of the mice were observed, and the results are as Figure 5 shown. Figure 5 Figure A in Figure 5 shows the body weight change of the mice within 15 days, and Figure B shows the food intake of the mice within 14 days. It can be seen from Figure 5 that the body weight change trend of the mice is normal; there is no difference in the food intake between all the blank control groups and the administration groups, indicating that monomeric compound 2 has no obvious acute toxic effect on the digestive system and does not affect energy metabolism and internal environment homeostasis.
[0079] The blood biochemistry of the mouse serum was detected to evaluate the effect on the liver and kidneys of the mice after administration. The results are shown in Figures A - C of Figure 6 , indicating that after two weeks of administration of the ointment containing 15% monomeric compound 2, the contents of aspartate aminotransferase (AST), uric acid (UA) and blood urea nitrogen (UREA) in the mice did not change significantly. Thus, it can be seen that topical administration of the cream containing 15% monomeric compound 2 will not cause liver damage.
[0080] The heart, liver, spleen, lungs, and kidneys of the mice were dissected, and the visceral-to-body weight ratio diagrams are as Figure 7 shown. It can be seen from the figure that there is little difference in the visceral-to-body weight ratio between the blank control group and the drug administration group, indicating that topical administration of the cream containing 15% of monomeric compound 2 does not cause physiological changes in the organs of the mice.
[0081] The skin of the mice was sectioned, and the results are as Figure 8 shown. The results show that topical administration of the cream containing 15% of monomeric compound 2 does not cause pathological changes or inflammation in the skin of the mice.
[0082] From the above data, it can be seen that topical administration of the cream containing 15% of monomeric compound 2 for treatment does not cause any pathological changes in the heart, liver, spleen, kidneys, lungs, and skin.
[0083] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.
Claims
1. Use of an extract from a plant of the Asteraceae family in the treatment and / or prevention of diseases associated with type 4 phosphodiesterase, wherein the extract from the Asteraceae family is an extract from Eclipta prostrata.
2. The use according to claim 1, characterized in that: The Eclipta prostrata extract is obtained by alcohol extraction of Eclipta prostrata, and the alcohol extraction method is as follows: weigh dry leaves of Eclipta prostrata, crush them into coarse powder, add the same volume of alcohol solution and soak them three times, wherein the first time is 2-5 days, and the second and third times are 1-3 days respectively, filter, combine the filtrate, reduce pressure to recover the effective components of the alcohol, extract the above recovered products with petroleum ether and ethyl acetate, reduce pressure to recover the effective components, and concentrate the remaining liquid to a thick paste to obtain the Eclipta prostrata alcohol extract containing the effective parts.
3. The use according to claim 1, characterized in that: Application of the wedelilactone compounds in the Asteraceae plant extract in the treatment and / or prevention of type 4 phosphodiesterase-related diseases.
4. The use according to claim 3, characterized in that: The wedeli lactone compound is one of compound 1 and compound 2 shown in the following formula I, the structural formula of compound 1 is 1 in the following formula I, and the structural formula of compound 2 is 2 in the following formula I; 5. A method for preparing a wedelilactone compound, characterized in that: The following steps are involved: A. crushing the dried leaves of Eclipta prostrata and soaking them in methanol or ethanol solution for three times, filtering, combining and collecting the filtrate, and then concentrating under reduced pressure to a paste to obtain an Eclipta prostrata alcohol extract; B. adding distilled water to the alcohol extract to prepare a suspension, extracting with petroleum ether and ethyl acetate in sequence, and concentrating the ethyl acetate extract under reduced pressure to obtain an ethyl acetate extract; C. The ethyl acetate extract is separated and purified by column chromatography to obtain monomer compound 1 and monomer compound 2.
6. The method for preparing the wedelilactone compound according to claim 5, characterized in that: The step C comprises: 1) The ethyl acetate extract was separated by silica gel column chromatography, and methanol-water gradient elution was performed at volume ratios of 5:95, 30:70, 55:45, 80:20 and 95:5, respectively, and the methanol-water eluates with volume ratios of 30:70 and 55:45 were collected; 2) taking the methanol-water eluates with a volume ratio of 30:70 and 55:45 for ODS column chromatography, gradient eluting with methanol-water with a volume ratio of 20:80, 30:70, 55:45, 65:35 and 95:5, and collecting and concentrating the methanol-water eluates with a volume ratio of 30:70 and 55:45 respectively; 3) The methanol-water eluate was purified by semi-preparative HPLC using methanol-water as the mobile phase in a volume ratio of (95-5):(5-95) to obtain Compound 1 and Compound 2.
7. An application in the preparation of a health product or food for treating and / or preventing psoriasis, wherein the health product or food comprises the wedelilactone compound according to claim 1.
8. A drug for treating and / or preventing psoriasis, characterized in that: The drug comprises one or both of the Asteraceae plant extract as described in claim 1 or the Wedelia lactone compound with any structure shown in claim 4, and pharmaceutically acceptable excipients and carriers.
9. The drug according to claim 8, characterized in that The dosage form of the medicine is capsule, tablet, pill, granule, granule, spray, emulsion and ointment.