Application of elephantopus scaber extract in preparation of medicine for treating atopic dermatitis

Abstract

The invention provides an application of elephantopus scaber extract in preparation of a medicine for treating atopic dermatitis, the extract is obtained by extracting and refining different parts of elephantopus scaber, and has the activity of resisting atopic dermatitis. The invention further provides treatment effects of different elephantopus scaber extracts on different effective parts, and industrial production is facilitated.

Description

Technical Field

[0001] The invention relates to an extract of Eleutherodactyla scabra and application thereof in atopic dermatitis, belonging to the field of medicine. Background Art

[0002] Atopic dermatitis (AD; also known as "atopic eczema") is the most common chronic, recurrent skin disease. Due to its high incidence, slow course, repeated attacks, and difficulty in curing, it has caused a lot of trouble for dermatologists and patients and has become a major public health problem worldwide. Although the exact pathogenesis of AD is still unclear, studies have shown that factors such as skin barrier dysfunction, immune abnormalities, and skin flora disorders are all related to the onset of AD.

[0003] The course of AD is complex and has obvious heterogeneity. No drug can cover all AD. However, due to the advantages of rich resources, definite efficacy, multiple pathways and targets, and few side effects, the development of new anti-atopic dermatitis drugs from natural plant resources has become a research hotspot.

[0004] The chemical components of the herb are rich, mainly including sesquiterpenoid lactones, triterpenoids, flavonoids, sterols and other ingredients. It is often used to treat various inflammatory diseases such as acute pharyngitis, acute gastroenteritis, acute otitis media, icteric hepatitis, nephritic edema, lung heat cough, urinary tract infection, bacillary dysentery, etc. It also has good effects in treating stomach problems, toothache, dysentery, hepatitis and other diseases.

[0005] Although it has been reported that Eleutherodactyla has anti-inflammatory effects, since the pathogenesis of AD itself is different from that of common inflammation, it is generally not considered to have a similar common mechanism with inflammation. Therefore, there have been no reports on the treatment of AD with Eleutherodactyla. Summary of the invention

[0006] According to the above technical research, the present invention provides an extract of Eleutherodactyla scabra and its application in preparing a drug for treating atopic dermatitis.

[0007] Furthermore, in the above application, the Eleutherodactyl extract is from the whole plant, stem, leaf or root of Eleutherodactyl.

[0008] Furthermore, in the above application, the Eleutherodactyla scabra extract is an ethanol extract or an effective part of the ethanol extract.

[0009] Furthermore, in the above application, the preparation method of the effective part of the extract of the spatholobiaceae comprises: after the ethanol extract is dispersed with water, it is extracted with an organic solvent A, the extract is collected, and it is concentrated to obtain extract 1 of a series of organic solvents A; or after the ethanol extract 1 is dispersed with water, it is adsorbed on a macroporous resin and eluted with a mobile phase B, the eluate is collected, and it is concentrated to obtain extract 2 of a series of mobile phases B; the organic solvent A is petroleum ether, ethyl acetate and n-butanol in sequence; the mobile phase B is water, an ethanol-water gradient solution with a volume fraction of 5-95%, an acetone-water gradient solution with a volume fraction of 5-95%, or a methanol-water gradient solution with a volume fraction of 5-95%.

[0010] Specifically include the following preparation methods:

[0011] 1) Weigh different parts of the herb, crush them, and then use an extraction solvent for cold soaking, the volume ratio of the raw material to the extraction solvent is 1:2-20, and filter and combine the solutions to obtain an extract;

[0012] 2) Concentration: The extract obtained in step 1) is concentrated to form an extract to obtain extract 1;

[0013] 3) Purification: After dispersing the extract 1 obtained in step 2) with water, extract it with organic solvent A, collect the extracts, and concentrate them to obtain extracts 2 of series organic solvent A; or after dispersing the extract 1 with water, adsorb it on a macroporous resin and elute it with mobile phase B, collect the eluates, and concentrate them to obtain extracts 3 of series mobile phase B, wherein the extracts 1, 2 and 3 are the extracts of the herbaceous spatholobi.

[0014] The present invention also provides an extract of Eleutherodactyla scabra, wherein the different parts of the Eleutherodactyla scabra are preferably: the whole plant, stems, leaves and roots, and more preferably the whole plant.

[0015] In the above step 1, different parts of the Euphorbia pulverata are preferably crushed into 10-40 meshes.

[0016] The present invention also provides an extract of Eleutherodactyla scabra, wherein the extraction solvent is a water-containing lower alcohol, i.e., various alcohols with a carbon number of less than 4, preferably an ethanol aqueous solution with a volume fraction of 75%-95%, and more preferably an ethanol aqueous solution with a volume concentration of 95%.

[0017] A patent discloses a method for extracting the total lactone compounds in the plant of the genus Eleutherodactyl, and uses an extraction solvent of 20%-95% ethanol water to extract the plant of the genus Eleutherodactyl. In the present invention, 75%-95% ethanol water is preferably used, and the purpose is to more comprehensively extract various polar compounds in the plant of the genus Eleutherodactyl, and is more inclined to extract relatively small polar compounds, rather than just focusing on the total lactone compounds in the plant of the genus Eleutherodactyl.

[0018] The present invention also provides an extract of Eleutherodactyla scabra, wherein the extraction method of the extraction solvent in step 1) is preferably a reflux extraction method.

[0019] In order to improve the effect of reflux extraction, the reflux extraction method is preferably: the extraction solvent is 95% ethanol aqueous solution, after the different parts of the spatholobus are crushed, the volume ratio of the different parts of the spatholobus to the extraction solvent is preferably 1:2-20, preferably 1:8-12; the reflux extraction time is 1.5-3 hours each time, preferably 2 hours, the extraction solvent temperature is: 78-88°C, three extractions are performed in total, filtered, and the filtrate is combined to obtain an extract.

[0020] In step 2), the extract obtained in step 1) is concentrated to form an extract, and the extract is concentrated until no liquid is left, thereby obtaining extract 1;

[0021] Technical requirements before extract 1 enters step 3): the extract 1 needs to be pretreated before entering step 3. If an organic solvent is used for extraction, the organic solvent needs to be removed from the extract 1 to ensure the extraction effect. If the extract 1 needs to be adsorbed using a macroporous resin or a polyamide resin, the organic solvent in the extract 1 needs to be removed to ensure that the initial gradient of the mobile phase is not affected.

[0022] In step 3), when the extract 1 is extracted with an organic solvent, in order to ensure the extraction effect and experimental cost, the volume ratio of the extract 1 after dispersion with water to the volume ratio of the organic solvent should be 1:1-3, preferably 1:1-1.5. During the extraction, the mobile phase A is required to extract one by one, and the extraction is concentrated after the extraction. The organic solvent A is petroleum ether, ethyl acetate and n-butanol in sequence.

[0023] After the organic phase A is extracted and concentrated, a series of extracts 2 of the organic phase A are obtained, which are also called the active parts of the organic phase A, namely, petroleum ether active parts, ethyl acetate active parts, n-butanol active parts and water active parts.

[0024] The type and sequence of the organic phase A are obtained by the inventors after extensive research and exploration, and the purpose is to obtain extracts with biological activity as much as possible.

[0025] In the specific embodiment, the active parts of petroleum ether and ethyl acetate are better than or close to the biological activity level of the above-mentioned extract 1 (i.e. 95% ethanol reflux extract of Herba Lycopodiellae) in biological activity. Other active part extracts also have biological activity, indicating that the active parts of petroleum ether, ethyl acetate have enriched the biologically active substances in Herba Lycopodiellae well, which is better than the biological activity effect of extract 1 (crude extract). The active parts are further preferably the active parts of petroleum ether and ethyl acetate.

[0026] The present invention further provides a Herba Eupatorii extract. In the extraction step of the extract, the mobile phase B is water, a 5-95% volume fraction ethanol-water gradient solution, a 5-95% volume fraction acetone-water gradient solution, or a 5-95% volume fraction methanol-water gradient solution.

[0027] When extract 1 is chromatographically separated by macroporous resin or polyamide resin, extract 1 needs to be dispersed in water for easy column adsorption. When open macroporous resin or is used, mobile phase B is used for elution, and after elution, it is concentrated to obtain extract 3.

[0028] The macroporous resin preferably has a filler of a macroporous resin of type D series, HPD series, HP series, XAD series or AB-8, preferably D101 series.

[0029] The mobile phase B is water, 5-95% by volume ethanol-water gradient solution, 5-95% by volume acetone-water gradient solution, or 5-95% by volume methanol-water gradient solution.

[0030] The mobile phase B is further preferably a gradient eluent of water, 20% ethanol aqueous solution, 50% ethanol aqueous solution, 70% ethanol aqueous solution, 90% ethanol aqueous solution and anhydrous ethanol.

[0031] In the above-mentioned process of eluting the macroporous resin using mobile phase B, the volume ratio of the column volume of the macroporous resin to the mobile phase B is 1:3-5.

[0032] After elution and concentration by mobile phase B, extract 3 of the mobile phase B is obtained, which is also called the active part of mobile phase B.

[0033] In the specific embodiment, research found that the macroporous resin 20% ethanol elution position group, the macroporous resin 50% ethanol elution position group and the macroporous resin 90% ethanol elution position group are better than or close to the biological activity level of the above-mentioned extract 1 (i.e. the 95% ethanol reflux extract of the Herba Lycopodii) in biological activity. And other elution position groups also have biological activity, indicating that the macroporous resin 20% ethanol elution position group, the macroporous resin 50% ethanol elution position group and the macroporous resin 90% ethanol elution position group have been enriched with physiologically active substances in the Herba Lycopodii, and are better than the biological activity effect of the extract 1 (crude extract). Preferred macroporous resin 50% ethanol elution position group and macroporous resin 90% ethanol elution position group. Wherein the biological activity effect that the macroporous resin 90% ethanol elution position has is the best.

[0034] In order to further specify the preparation process of the Herba Lycopodii extract, the present invention further provides a Herba Lycopodii extract, wherein the extract is prepared by the following steps:

[0035] 1) Weigh the dried whole herb of Euphorbia obesa, crush it into 20 mesh, use 95% ethanol aqueous solution for reflux extraction, the solid-liquid volume ratio is 1:10, reflux extraction for 2 hours each time, extract 3 times in total, the extraction temperature is 78-88° C., filter and combine the filtrate to obtain an extract;

[0036] 2) Concentration: The extract is concentrated to an extract to obtain an extract 1, wherein water is removed from the extract 1;

[0037] 3) Purification: After dispersing the extract 1 with water, extracting with organic phase A, extracting with petroleum ether, dichloromethane, ethyl acetate and n-butanol in sequence, the volume of each component of organic phase A is 1-1.5 times the volume of the extract 1 after dispersing with water, collecting the eluent, and concentrating them to obtain a series of organic phase extracts 2; or the extract is prepared by the following steps:

[0038] 1) Weigh the dried whole herb of Euphorbia cerifera, crush it into 20 meshes, reflux it with 95% ethanol aqueous solution, filter and combine the filtrate to obtain an extract;

[0039] 2) Concentration: The extract is concentrated to an extract to obtain an extract 1, wherein the organic solvent is removed from the extract 1;

[0040] 3) Refining: After extract 1 is dispersed with a small amount of water, it is completely adsorbed on a macroporous resin D101 column and eluted with mobile phase B, wherein the mobile phase B is water, 20% ethanol aqueous solution, 50% ethanol aqueous solution, 70% ethanol aqueous solution, 90% ethanol aqueous solution and anhydrous ethanol eluent, respectively, and the volume of each mobile phase component is 3-5 times the volume of the macroporous resin D101 column. The eluents of each part are collected and concentrated to obtain a series of mobile phase extracts 3. The obtained series of mobile phase extracts 3 are also called the active sites of mobile phase B.

[0041] The technical requirement of the small amount of water in the above-mentioned macroporous resin technical solution is: the extract 1 can be dispersed to facilitate the column adsorption operation, but the amount of water cannot affect the elution gradient effect of the mobile phase B during elution, which is a common operating technique for personnel in this field.

[0042] In the macroporous resin extraction technology solution, the extraction method of the dried whole herb in step 1) is preferably reflux extraction with 95% ethanol aqueous solution.

[0043] In the above-mentioned Herba Eupatorii extract, whether liquid-liquid extraction or macroporous resin chromatography is adopted, the concentration in steps 2) and 3) is preferably concentrated under reduced pressure. The extract is concentrated to an extract until no liquid is spun out.

[0044] When the extract 1 is chromatographed using a macroporous resin, the mobile phase B is water, 20% ethanol aqueous solution, 50% ethanol aqueous solution, 70% ethanol aqueous solution, 90% ethanol aqueous solution and anhydrous ethanol eluent, respectively, and the volume of each mobile phase component is 3-5 times the column volume of the macroporous resin D101, the active sites of the obtained mobile phase B have the following characteristics:

[0045] The active part of the extract of Eleutherodactylum scabra was eluted with macroporous resin and water, and the main components were polysaccharides and polyphenols. The developing solvent of TLC was dichloromethane:methanol=3:1. f The value is less than 0.1.

[0046] The active part of the extract of the scutellaria baicalensis was eluted with 20% ethanol in the macroporous resin, which mainly contained flavonoid glycosides and phenolic glycosides. Under the condition of TLC detection with dichloromethane:methanol=3:1 as the developing solvent, R f value f The value is 0.2-0.5.

[0047] The active part of the extract of the scutellaria baicalensis was eluted with 50% ethanol and mainly contained caffeic acid compounds. The developing solvent of TLC detection was petroleum ether: acetone = 3:1 under the condition of R f The value is 0.3-0.7.

[0048] The active part of the extract of the scutellaria baicalensis was eluted with 70% ethanol in macroporous resin, which mainly contained ionones and sterols. The developing solvent of TLC detection was petroleum ether: acetone = 5:1 under the condition of R f The value is 0.2-0.5.

[0049] The active part of the extract of the scutellaria baicalensis was eluted with 90% ethanol and anhydrous ethanol. The main components were sesquiterpene lactone compounds. Under the condition of TLC detection with petroleum ether: ethyl acetate = 6:1, R f The value is 0.4-0.7.

[0050] It can be seen that the active parts of the extracts contain different components and therefore have different biological activities.

[0051] The invention provides an extract of Eleutherodactyla scabra, which is comprehensively and meticulously extracted from Eleutherodactyla scabra. The extract is a good supplement to the deficiencies of existing drugs and is also an important guide to the current extracts and extraction processes of Eleutherodactyla scabra.

[0052] Another object of the present invention is to provide an anti-specific dermatitis composition, which is composed of an effective dose of the Herba Lysimachiae extract of the present invention and a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient refers to an excipient required for making any pharmaceutical dosage form suitable for human or animal use. For example, when making an oral solid preparation, the pharmaceutically acceptable excipient refers to a diluent, a binder, a wetting agent, a disintegrant, a lubricant, and a glidant; when making an injection, the pharmaceutically acceptable excipient refers to a pH regulator, a cosolvent, an antioxidant, an isotonic agent, etc. The drug refers to a tablet, a pill, a capsule, a suspension, an ointment or an emulsion, etc.

[0053] The results of in vitro or in vivo pharmacodynamic tests show that the Eleutherodactylus scabra plant extract of the present invention has anti-atopic dermatitis activity, and different concentrations of Eleutherodactylus scabra extract can show obvious improvement effects on atopic dermatitis mice induced by MC903 and OXA.

[0054] Therefore, the present invention further provides the use of the above-mentioned Eleutherodactylum japonicum extract in the preparation of anti-atopic dermatitis drugs.

[0055] The inventors verified the beneficial effects of the Eleutherodactyla extract and different active parts of the present invention through the following experiments.

[0056] It is reiterated that the following experiments are only exemplary experiments among the many experiments conducted during the development of the present invention, and do not cover and exhaust all the experiments conducted by the inventor for the present invention. The purpose is only to use those data to illustrate the anti-atopic dermatitis activity of the Eleutherococcus scoparia extract and active parts of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 :Typical photos of 85-HP-90 sites on MC903-induced atopic dermatitis in mice ears;

[0058] Figure 2 : Representative photos of 85-HP-90 sites on OXA-induced atopic dermatitis in mouse ears. DETAILED DESCRIPTION

[0059] Example 1

[0060] Preparation of 95% ethanol cold-infusion extract of Herba Edulis

[0061] 20 kg of the whole dried grass of Eleutherodactyla was crushed and extracted with 12 times the volume of 95% ethanol (temperature: 20-25°C) for 3 times, each time for 7 days, the extracts were combined, filtered, and the filtrate was concentrated under reduced pressure to obtain 0.6 kg of 95% ethanol extract, which was the ethanol extract.

[0062] Example 2

[0063] Preparation of 95% ethanol extract of physalis edulis from petroleum ether

[0064] The 95% ethanol extract obtained in Example 1 was dispersed with 5 L of distilled water, and 7.5 L of petroleum ether was added for extraction. After the solvent was recovered, 100 g of the petroleum ether fraction was obtained, which was marked as 95-P fraction.

[0065] Example 3

[0066] Sample preparation of ethyl acetate layer of 95% ethanol extract of Euphorbia milii

[0067] The sample extracted in Example 2 was further extracted with ethyl acetate, and after the solvent was recovered, 140 g of the ethyl acetate fraction was obtained, which was marked as fraction 95-E.

[0068] Example 4

[0069] Sample preparation of n-butanol layer of 95% ethanol extract of Euphorbia milii

[0070] The sample extracted in Example 4 was further extracted with n-butanol, and after the solvent was recovered, 145 g of n-butanol fraction was obtained, which was marked as fraction 95-B.

[0071] Example 5

[0072] Preparation of aqueous layer sample of 95% ethanol extract of Euphorbia milii

[0073] The remaining part after the extraction in Example 4 was directly used to recover the solvent to obtain 200 g of a water fraction, which was marked as the 95-W fraction.

[0074] Example 6

[0075] Preparation of 95% ethanol reflux extract of Eleutherococcus chinensis

[0076] 20 kg of the whole dried herb of Eleutherococcus scabra was crushed and extracted with 12 times the volume of 95% ethanol under heating reflux (80°C) for 3 times, each time for 2 hours. The extracts were combined, filtered, and the filtrate was concentrated under reduced pressure to obtain 1.4 kg of 95% ethanol extract.

[0077] Example 7

[0078] Sample preparation of the water-eluted fraction of the macroporous adsorption resin of 95% ethanol extract of Euphorbia milii

[0079] 1.4 kg of 95% ethanol extract of Eleutherodendron edulis was dispersed in 8 L of water and then adsorbed on 20 L of HP-20 macroporous adsorption resin, eluted with distilled water for 3 column volumes, and the eluate was collected and concentrated under reduced pressure to obtain 453 g of 95% ethanol extract of Eleutherodendron edulis macroporous adsorption resin water elution fraction, which was marked as 95-HP-0 fraction.

[0080] Example 8

[0081] Sample preparation of the 95% ethanol extract of Euphorbia elegans by macroporous adsorption resin eluted with 20% ethanol

[0082] The macroporous adsorption resin column eluted in Example 7 was eluted with 20% ethanol, and after concentrating and recovering the solvent, 219 g of the 95% ethanol extract of Eleutherodactylum sinensis macroporous adsorption resin 20% ethanol elution fraction was obtained, which was marked as 95-HP-20 fraction.

[0083] Example 9

[0084] Sample preparation of the 50% ethanol elution fraction of the macroporous adsorption resin of the 95% ethanol extract of Euphorbia milii

[0085] The macroporous adsorption resin column eluted in Example 8 was eluted with 50% ethanol, and after concentrating and recovering the solvent, 185 g of the 50% ethanol elution fraction of the 95% ethanol extract of Eleutherodactylum sinensis was obtained, which was marked as the 95-HP-50 fraction.

[0086] Example 10

[0087] Sample preparation of the 95% ethanol extract of Euphorbia elegans by macroporous adsorption resin eluted with 70% ethanol

[0088] The macroporous adsorption resin column eluted in Example 9 was eluted with 70% ethanol, and after concentrating and recovering the solvent, 144 g of the 95% ethanol extract of Eleutherodactylum sinensis macroporous adsorption resin 70% ethanol elution fraction was obtained, which was marked as 95-HP-70 fraction.

[0089] Embodiment 11

[0090] Sample preparation of the 90% ethanol elution fraction of the 95% ethanol extract of Euphorbia milii macroporous adsorption resin

[0091] The macroporous adsorption resin column eluted in Example 10 was eluted with 90% ethanol, and after concentrating and recovering the solvent, 55 g of the 90% ethanol elution fraction of the 90% ethanol extract of Eleutherodactylum sinensis was obtained, which was marked as the 95-HP-90 fraction.

[0092] Example 12

[0093] Preparation of the sample of the 95% ethanol extract of Eleutherodendron macroporous adsorption resin fraction eluted with anhydrous ethanol The macroporous adsorption resin column eluted with anhydrous ethanol after Example 11 was eluted with anhydrous ethanol, and after concentrating and recovering the solvent, 285 g of the fraction eluted with anhydrous ethanol extract of Eleutherodendron macroporous adsorption resin was obtained, which was marked as the fraction 95-HP-100.

[0094] Embodiment 13

[0095] Preparation of 85% ethanol reflux extract of Eleutherococcus chinensis

[0096] 20 kg of the dried whole herb of Eleutherodactyla was crushed and extracted twice with 15 times the volume of 85% ethanol under heating reflux (80°C), each time for 2 hours. The extracts were combined, filtered, and the filtrate was concentrated under reduced pressure to obtain 1.6 kg of 85% ethanol extract.

[0097] Embodiment 14

[0098] Sample preparation of the water-eluted fraction of the macroporous adsorption resin of 85% ethanol extract of Euphorbia milii

[0099] 1.6 kg of 85% ethanol extract of Eleutherodendron edulis was dispersed in 8 L of water and then adsorbed on 20 L of AB-8 macroporous adsorption resin, eluted with distilled water for 3 column volumes, and the eluate was collected and concentrated under reduced pressure to obtain 570 g of the macroporous adsorption resin water elution fraction of 85% ethanol extract of Eleutherodendron edulis, which was marked as 85-HP-0 fraction.

[0100] Embodiment 15

[0101] Sample preparation of the 20% ethanol elution fraction of the macroporous adsorption resin of the 85% ethanol extract of Euphorbia milii

[0102] The macroporous adsorption resin column eluted from Example 14 was eluted with 20% ethanol, and after concentrating and recovering the solvent, 208 g of the 95% ethanol extract of Eleutherodactylum sinensis macroporous adsorption resin 20% ethanol elution fraction was obtained, which was marked as 85-HP-20 fraction.

[0103] Example 16

[0104] Sample preparation of the 50% ethanol elution fraction of the macroporous adsorption resin of the 85% ethanol extract of Euphorbia milii

[0105] The macroporous adsorption resin column eluted from Example 15 was eluted with 50% ethanol, and after concentrating and recovering the solvent, 192 g of the 50% ethanol elution fraction of the macroporous adsorption resin of the 85% ethanol extract of Eleutherodactylum scabra was obtained, which was marked as the 85-HP-50 fraction.

[0106] Embodiment 17

[0107] Sample preparation of the 85% ethanol extract of Euphorbia elegans by macroporous adsorption resin eluted with 70% ethanol

[0108] The macroporous adsorption resin column eluted with Example 16 was eluted with 70% ethanol, and after concentrating and recovering the solvent, 135 g of the 70% ethanol elution fraction of the 85% ethanol extract of Eleutherodactylum sinensis was obtained, which was marked as the 85-HP-70 fraction.

[0109] Embodiment 18

[0110] Sample preparation of the 90% ethanol elution fraction of the macroporous adsorption resin of the 85% ethanol extract of Euphorbia milii

[0111] The macroporous adsorption resin column eluted in Example 17 was eluted with 90% ethanol, and after concentrating and recovering the solvent, 78 g of the 90% ethanol elution fraction of the 90% ethanol extract of Eleutherodactylum sinensis was obtained, which was marked as the 85-HP-90 fraction.

[0112] Embodiment 19

[0113] Sample preparation of the 85% ethanol extract of Euphorbia milii by macroporous adsorption resin eluted with anhydrous ethanol

[0114] The macroporous adsorption resin column eluted in Example 18 was eluted with anhydrous ethanol, and after concentrating and recovering the solvent, 294 g of the anhydrous ethanol elution fraction of the macroporous adsorption resin of the anhydrous ethanol extract of Eleutherodactylum scabra was obtained, which was marked as the 85-HP-100 fraction.

[0115] Embodiment 20

[0116] Preparation of 75% ethanol reflux extract of Herba Edulis

[0117] 20 kg of the dried whole herb of Eleutherodactyla was crushed and extracted twice with 15 times the volume of 75% ethanol under heating reflux (80°C), each time for 2 hours. The extracts were combined, filtered, and the filtrate was concentrated under reduced pressure to obtain 1.8 kg of 75% ethanol extract.

[0118] Embodiment 21

[0119] Preparation of 75% ethanol extract of physalis edulis from petroleum ether

[0120] The 75% ethanol extract obtained in Example 20 was dispersed with 10 L of distilled water, and 10 L of petroleum ether was added for extraction. After the solvent was recovered, 250 g of the petroleum ether fraction was obtained, which was marked as the 75-P fraction.

[0121] Embodiment 22

[0122] Sample preparation of ethyl acetate layer of 75% ethanol extract of Euphorbia pulegosa

[0123] The sample extracted in Example 21 was further extracted with ethyl acetate, and after the solvent was recovered, 340 g of the ethyl acetate fraction was obtained, which was marked as fraction 75-E.

[0124] Embodiment 23

[0125] Sample preparation of n-butanol layer of 75% ethanol extract of Euphorbia milii

[0126] The sample extracted in Example 22 was further extracted with n-butanol, and after the solvent was recovered, 580 g of n-butanol fraction was obtained, which was marked as fraction 75-B.

[0127] Embodiment 24

[0128] Preparation of aqueous layer sample of 75% ethanol extract of Euphorbia pulegosa

[0129] The remaining part after the extraction of Example 23 was directly used to recover the solvent to obtain 600 g of water fraction, which was marked as 75-W fraction.

[0130] Embodiment 14

[0131] Experimental study on the anti-atopic dermatitis activity of the extract of Eleutherodactylum scabra and its active fraction

[0132] 1. Effects on the ears of mice with MC903-induced atopic dermatitis

[0133] BALB / c mice, SPF grade, weighing 18-22g, were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., license number: SCXK (Beijing) 2016-0009. Mice were group-housed 6 per cage, with free access to food and water, room temperature of 23-25℃, humidity of 50±10%, and light from 7:00 to 19:00. The experiment began after the animals adapted to the new environment for 3 days. On the morning of the formal experiment, except for the mice in the normal control group, 0.1mM MC903 (dissolved in anhydrous ethanol) was applied to the left and right ears of each mouse, 10μL / ear; 10 minutes later, 10μL of anhydrous ethanol was applied to the left and right ears of the mice in the normal control group and the model group; 10μL of anhydrous ethanol was applied to the left ear of the mice in the drug administration group, and 10μL of 5% crude product of local gall grass (dissolved in anhydrous ethanol) was applied to the right ear. This operation was performed once a day, and the growth status of the mice was monitored during the period. After 14 days, the severity of the skin lesions of the mice was observed, and the mice were euthanized. Ear pieces with a diameter of 8 mm were taken from the same position of the left and right ears and weighed.

[0134] The results are shown in Table 1: After the modeling agent MC903 was administered, the skin of the mouse ears was severely damaged, with typical atopic dermatitis symptoms such as obvious edema, erythema, epidermal exfoliation, scab, lichenification and dryness. The improvement of atopic dermatitis by administering crude products of Eleutherodendron sibiricum with different processes was quite different (Table 1), indicating that the separation effect was obvious; among them, 85-HP-90 (90% elution site) had the best effect ( Figure 1 ).

[0135] Table 1 Effects of the extract of Eleutherodactylum japonicum and its active fraction on the ear weight of mice with atopic dermatitis induced by MC903 (n=6; P<0.05 was considered to be significantly different)

[0136]

[0137]

[0138] Typical mouse improvement pictures are shown in Figure 1 .

[0139] 2. Effects on the ears of mice with OXA-induced atopic dermatitis

[0140] The mice were randomly divided into groups, with 6 mice in each group. On the first day, in addition to the normal control group mice being coated with 10μl acetone-olive oil, each mouse was sensitized by applying 10μL 4% OXA to the left and right ears. Starting from the 7th day, in addition to the normal control group mice being coated with 10μL acetone-olive oil, each mouse was attacked by applying 10μL 2% OXA to the left and right ears, and the attack was carried out once every 2 days. From the beginning of sensitization, the test substance was administered once a day, and the growth status of the mice was monitored during this period. After 25 days, the severity of the skin lesions of the mice was observed, the mice were euthanized, and the ear pieces with a diameter of 8mm were taken from the same position of the left and right ears and weighed.

[0141] The results are shown in Table 2: After the modeling agent OXA was administered, the mouse ear skin was severely damaged, with obvious edema, erythema, epidermal exfoliation, crusting, lichenification and dryness, and other typical atopic dermatitis symptoms. The effects of different processes of the extracts of the herb Eleutherodactylum on atopic dermatitis were very different (Table 2), indicating that the separation effect was obvious; among them, 85-HP-90 (90% elution site) had the best effect ( Figure 2 ), which is consistent with the performance on the MC903 model.

[0142] Table 2 Effects of the extract of Herba Lysimachiae and its active fraction on the ear weight of mice with OXA-induced atopic dermatitis (n=6; P<0.05 was considered to be significantly different)

[0143]

[0144]

[0145] Typical mouse pictures are shown in Figure 2 .

Claims

1. Use of a Herba Lysimachiae extract in the preparation of a drug for treating atopic dermatitis.

2. The application according to claim 1, characterized in that: The Eleutherodactyl extract is from the whole stem, leaf or root of Eleutherodactyl.

3. The application according to claim 1, characterized in that: The Eleutherodactylum japonicum extract is an ethanol extract or an effective fraction of the ethanol extract.

4. The use according to claim 3, characterized in that: The preparation method of the effective part of the extract of the scutellaria baicalensis comprises: dispersing the ethanol extract with water, extracting with an organic solvent A, collecting the extracts, and concentrating them to obtain extracts 1 of a series of organic solvents A; or dispersing the ethanol extract 1 with water, adsorbing it on a macroporous resin, eluting it with a mobile phase B, collecting the eluates, and concentrating them to obtain extracts 2 of a series of mobile phases B; the organic solvent A is petroleum ether, ethyl acetate, and n-butanol in sequence; and the mobile phase B is water, an ethanol-water gradient solution with a volume fraction of 5-95%, an acetone-water gradient solution with a volume fraction of 5-95%, or a methanol-water gradient solution with a volume fraction of 5-95%.

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