CN108114061B

CN108114061B - Use of Chinese herbal medicine extract and pharmaceutical composition containing the same and steroid

Info

Publication number: CN108114061B
Application number: CN201611205417.7A
Authority: CN
Inventors: 潘一红; 温淑芳; 吕居勋; 姚心然; 庄凯安; 林士弘; 李明翰; 谢岱儒; 许博尧; 吴信颉; 吕亦晃
Original assignee: Industrial Technology Research Institute ITRI
Current assignee: Industrial Technology Research Institute ITRI
Priority date: 2016-11-30
Filing date: 2016-12-21
Publication date: 2022-01-25
Anticipated expiration: 2036-12-21
Also published as: JP6738787B2; JP2018090568A; CN108114061A; TWI610679B; TW201821095A

Abstract

The invention provides an application of a Chinese herbal medicine extract in preparing a medicament for treating skin diseases, wherein the Chinese herbal medicine extract is prepared from raw materials of liquorice, radix bupleuri, scutellaria baicalensis, schisandra chinensis and Chinese herbaceous peony.

Description

Use of Chinese herbal medicine extract and pharmaceutical composition containing the same and steroid

Technical Field

The invention relates to application of a Chinese herbal medicine extract and a pharmaceutical composition containing the Chinese herbal medicine extract and steroids.

Background

Skin disorders are the most common disorders worldwide, with about 1/3 people presenting a pathological skin problem in their lives, and with health care costs, skin-related medical costs as high as 25%.

Skin diseases fall into four broad categories, dermatitis (e.g., allergic and contact), cancer (e.g., melanoma), immune diseases (e.g., psoriasis), and infectious skin diseases (e.g., bacterial, fungal, and viral infections), respectively.

Topical steroids have been widely used in a variety of skin conditions, particularly atopic dermatitis and psoriasis, and in severe dermatitis patients, the steroid doses are higher. Many compounds classified as steroids (steroids), such as betamethasone or prednisolone, are very effective in treating inflammatory diseases, however, they may also cause atrophy of the skin of patients after long-term use. Patients are generally considered steroid responders if skin atrophy occurs during steroid therapy. The effects of skin atrophy on patients already having symptoms of skin diseases (e.g., psoriasis patients) are of concern. However, even normal skin, under the condition of long-term use of steroids, side effects of skin damage occur.

Therefore, the development of safe alternatives to steroids and/or how to maintain the efficacy of topical steroid therapy and reduce side effects is an important issue.

Disclosure of Invention

The invention also provides the use of a Chinese herbal medicine extract for preparing a medicament for improving side effects of steroid treatment of skin diseases, wherein the Chinese herbal medicine extract is prepared from raw materials including licorice, bupleurum, scutellaria, schisandra and peony.

The present invention further provides a pharmaceutical composition for alleviating skin disorders, comprising: chinese medicinal extract, wherein the Chinese medicinal extract is prepared from Glycyrrhrizae radix, bupleuri radix, Scutellariae radix, fructus Schisandrae chinensis and radix Paeoniae; and a steroid, wherein the herbal extract has the effect of reducing side effects caused by the steroid and/or the effect of reducing the effective dose of the steroid required as compared to the use of the steroid alone.

Drawings

Fig. 1A shows photographs of the skin of mice in the control, control and four experimental groups in an Imiquimod (IMQ) -induced psoriasis-like animal model experiment.

FIG. 1B shows gene expression levels of IL-17A, IL-6, TNF- α, and S100A7 in mice from control, and four experimental groups in an imiquimod-induced psoriasis-like animal model experiment.

FIG. 2A shows photographs of the skin of mice in the control group and three mut mut mutexperimental groups (PT-A (100mg) ointment-treated group, PT-A-1(100mg) ointment-treated group, PT-A-2(100mg) ointment-treated group) in an imiquimod-induced psoriasis-like animal model mut mut mutexperiment.

FIG. 2B shows the desquamation plus redness severity scores of mice from control and three mut mut mutexperimental groups (PT-A (100mg) ointment treated group, PT-A-1(100mg) ointment treated group, PT-A-2(100mg) ointment treated group) in an imiquimod-induced psoriasis-like animal model mut mut mutexperiment.

FIG. 3 shows photographs of the skin of mice in the control group, control group and mutexperimental group (PT-A-DS (100mg/kg) intraperitoneal injection treatment group) and the result of hematoxylin-eosin (hematoxylin and eosin) staining of skin samples in an imiquimod-induced psoriasis-like animal model mutexperiment.

Fig. 4A shows photographs of the skin of mice of the control group, the control group and the two mut mutexperimental groups (PT-a (10mg) ointment-treated group and PT-a (100mg) ointment-treated group) in an imiquimod-induced psoriasis-like animal model mut mutexperiment.

FIG. 4B shows the gene mut mutexpression levels of IL-17A, IL-23 and IL-22 in mice from control, control and two mut mutexperimental groups (PT-A (10mg) ointment treated and PT-A (100mg) ointment treated) in an imiquimod-induced psoriasis-like animal model mut mutexperiment.

FIG. 4C shows the results of immunohistological staining with CD68, CD4, Neutrophil, Ki-67, and CD31 antibodies in mice from the control and mutexperimental groups (PT-A (100mg) ointment treated groups) in an imiquimod-induced psoriasis-like animal model mutexperiment.

FIG. 5A shows the increased thickness of the ears of mice in the control, positive control and mutexperimental groups (PT-A (50mg) ointment treated groups) in the 1-fluoro-2,4-dinitrobenzene (1-fluoro-2, 4-dinobenzene, DNFB) induced animal model mutexperiment for contact dermatitis.

FIG. 5B shows the results of CD4 immunohistological stain analysis and photographs of skin samples of inflamed portions of mice in the control, positive control and mutexperimental groups (PT-A (50mg) ointment treated group) in a 1-fluoro-2, 4-dinitrobenzene-induced animal contact dermatitis model mutexperiment.

FIG. 6A shows photographs of the skin of mice in the control group, control group and three mutexperimental groups (PT-A (100mg) ointment-treated group, Rinderon-V (0.06%, Taiwan salt wild, China) ointment-treated group and Daivon mutex (0.005%, LEO Pharma) ointment-treated group) in the imiquimod-induced psoriasis-like animal model mutexperiment.

FIG. 6B shows the gene mutexpression levels of IL-17A, IL-6, TNF- α, and S100A7 in mice from control, and three mutexperimental groups (PT-A (100mg) ointment treatment group, Rinderon-V (0.06%, Taiwan salt wild sense in China) ointment treatment group, and Daivon mutex (0.005%, LEO Pharma) ointment treatment group) in an imiquimod-induced psoriasis-like animal model mutexperiment.

Figure 7A shows photographs of the skin of mice from control (IMQ), control (IMQ + vehicle) and three mutexperimental groups (IMQ + BD (steroid) ointment treated, IMQ + BD + PT-a ointment treated (25mg) and daivobet (leo pharma) ointment treated) in an imiquimod-induced psoriasis-like animal model mutexperiment.

Figure 7B shows the desquamation plus redness severity scores for mice from control, control and three mutexperimental groups (BD (steroid) ointment treated, BD + PT-a ointment treated (25mg) and daivobet (leo pharma) ointment treated) in an imiquimod-induced psoriasis-like animal model mutexperiment.

Fig. 7C shows hematoxylin-eosin (hematoxylin and eosin) staining results of mouse skin samples of control, control and three mutexperimental groups (BD (steroid) ointment treatment group, BD + PT-a ointment treatment group (25mg) and daivobet (leo pharma) ointment treatment group) in an imiquimod-induced psoriasis-like animal model mutexperiment.

FIG. 7D shows the results of immunohistological staining with Ki-67 antibody in imiquimod-induced psoriasis-like animal model mutexperiments with control, control and three mutexperimental groups (BD (steroid) ointment treated group, BD + PT-A ointment treated group (25mg) and Daivobe (LEO Pharma) ointment treated group).

FIGS. 8A-1 and 8A-2 show photographs of the skin of mice of the control group, and five mut mut mut mut mutexperimental groups (fluocinonide (0.05%; mesosteroid) ointment-treated group, fluocinonide + PT-A (0.25%) ointment-treated group (PT-A to fluocinonide weight ratio of 5:1), fluocinonide + PT-A (5%) ointment-treated group (PT-A to fluocinonide weight ratio of 100:1), PT-A (5%) ointment-treated group, and clobetasol propionate (0.05%) ointment-treated group).

FIGS. 8B-1 and 8B-2 show hematoxylin-eosin staining results of skin samples of mice of the control group, and five mut mut mut mut mutexperimental groups (fluocinolone acetonide (0.05%; mesosteroid) ointment treatment group, fluocinolone acetonide + PT-A (0.25%) ointment treatment group (PT-A to fluocinolone weight ratio of 5:1), fluocinolone + PT-A (5%) ointment treatment group (PT-A to fluocinolone weight ratio of 100:1), PT-A (5%) ointment treatment group, and clobetasol propionate (0.05%) ointment treatment group).

Figure 8C shows the severity scores for redness, desquamation and skin thickness of mice from control, control and five mut mut mut mut mutexperimental groups (fluocinolone (0.05%; mesosteroid) ointment treatment, fluocinolone + PT-a (0.25%) ointment treatment (5: 1 wt PT-a to fluocinolone), fluocinolone + PT-a (5%) ointment treatment (100: 1 wt PT-a to fluocinolone), PT-a (5%) ointment treatment and clobetasol propionate (0.05%) ointment treatment).

FIG. 9A shows the ear thickness of mice from control group (vehicle) and three mutexperimental groups (PT-A (content 25mg/g) ointment treatment group, CLOBETASOL Cream (CLOBETASOL Cream, mixed Cream base CLOBETASOL propionate content 0.25mg/g) treatment group, and Koujiu hydrophilic ointment (TOPSYM Cream, mixed Cream base fluocinolone acetonide content 0.25mg/g) treatment group).

FIG. 9B shows the hematoxylin-eosin staining results of skin samples of mice of control group (vehicle) and three mutexperimental groups (PT-A (content 25mg/g) ointment treatment group, CLOBETASOL Cream (CLOBETASOL Cream, mixed Cream base CLOBETASOL propionate content 0.25mg/g) treatment group and Koufujing hydrophilic ointment (TOPSYM Cream, mixed Cream base fluocinolone content 0.25mg/g) treatment group).

FIG. 10A shows the ear thickness of mice from control (vehicle), control (Clobetasol) and two mut mut mutexperimental groups (PT-A (content 25mg/g) ointment treated group mixed with PT-A (content 25mg/g) Clobetasol Cream (content 0.25mg/g of Clobetasol Cream) (ointment treated group of Clobetasol + PT-A)).

FIG. 10B shows the hematoxylin-eosin staining results of skin samples from mice of control (vehicle), control (Clobetasol), and two mut mut mutexperimental groups (PT-A (content 25mg/g) ointment treated group mixed with PT-A (content 25mg/g) Clobetasol Cream (content 0.25mg/g of CLOBetasol Cream) (ointment treated group of Clobetasol + PT-A)).

FIG. 11A shows the appearance of skin in mice from control (no treatment), control (Rinderon-V (0.06%) ointment treated), and mutexperimental (PT-A (25 mg/g content) ointment treated groups.

FIG. 11B shows the hematoxylin-eosin staining results of skin samples of mice of the control group (without any treatment), the control group (Rinderon-V (0.06%) ointment-treated group), and the mutexperimental group (PT-A (content 25mg/g) ointment-treated group).

FIG. 12 shows the results of repeated irritation and sensitization tests on PT-A (25 mg/g) ointment treated human skin. In the human species column, C represents Caucasian (Caucasian), H represents latin american (Hispanic); in the reaction field, dc (uncontinuous) means that the subject is absent from the test; in the score column, N/A (not applicable) represents that the score could not be calculated due to the absence of the subject.

Detailed description of the preferred embodiments

In one embodiment of the present invention, there is provided a use of a Chinese herbal medicine extract for the preparation of a medicament for treating skin diseases.

The skin disease may include an autoimmune-related skin disease or an infectious skin disease. Examples of the above autoimmune-related skin diseases include, but are not limited to, psoriasis, allergic dermatitis, atopic dermatitis, etc.

In the use of the Chinese herbal medicine extract for preparing a medicament for treating skin diseases, the Chinese herbal medicine extract can be prepared from raw materials including licorice, radix bupleuri, radix scutellariae, schisandra chinensis and peony. In one embodiment, the Chinese herbal extracts are prepared from raw materials comprising licorice, bupleurum, scutellaria, schisandra and peony.

Examples of the above-mentioned licorice may include raw licorice, honey-fried licorice, etc., but are not limited thereto. The above mentioned bupleurum root may include, but is not limited to, raw bupleurum chinense and high bupleurum chinense. The scutellaria may include, for example, scutellaria cumicifolia, scutellaria barbata, etc., but is not limited thereto. Examples of the aforementioned schisandra may include, but are not limited to, schisandra chinensis, etc. In addition, the peony can include, but is not limited to, red peony, white peony, and the like.

In the preparation raw materials of the Chinese herbal medicine extract, the weight ratio of the liquorice, the radix bupleuri, the scutellaria baicalensis, the schisandra chinensis and the peony can be about 1-5: 1-5: 1-5: 1-5: 1-5, but is not limited thereto. In one embodiment, in the raw materials for preparing the Chinese herbal medicine extract, the weight ratio of the licorice, the bupleurum, the scutellaria, the schisandra and the peony may be about 1:1:1:1: 1.

the method for obtaining the above-mentioned Chinese herbal medicine extract is not particularly limited, and can be obtained in various ways currently known in the art. In one embodiment, it is obtainable by an extraction process comprising the steps of: firstly, liquorice, radix bupleuri, scutellaria baicalensis, schisandra chinensis and Chinese herbaceous peony in the preparation raw materials are mixed to form a raw material mixture, and then the raw material mixture is subjected to an extraction process by a solvent. In another embodiment, it is obtained by another extraction method comprising the following steps: firstly, licorice, bupleurum, scutellaria, schisandra and peony in the above raw materials are extracted with solvent respectively to generate licorice extract, bupleurum extract, scutellaria extract, schisandra extract and peony extract, and then the licorice extract, bupleurum extract, scutellaria extract, schisandra extract and peony extract are mixed.

The solvent used for the extraction process may include water or an organic solvent, but is not limited thereto. The organic solvent may include, but is not limited to, alcohols, ketones, acids, esters, and the like. Examples of the alcohols include, but are not limited to, methanol, ethanol, and the like. In one embodiment, the solvent may be ethanol. In a specific embodiment, the solvent may be 30% ethanol. In another particular embodiment, the solvent can be 75% ethanol.

In one embodiment, in the raw materials for preparing the above Chinese herbal medicine extract, the licorice is raw licorice, the bupleurum is bupleurum chinense, the scutellaria baicalensis is calcined scutellaria baicalensis, the schisandra chinensis is schisandra chinensis, and the peony is red peony root. The weight ratio of the raw licorice, the bupleurum chinense, the dried scutellaria baicalensis, the schisandra chinensis and the red peony root can be about 1:1:1:1:1, but is not limited thereto. In another specific embodiment, the raw materials of the Chinese herbal medicine extract include raw licorice, bupleurum chinense, dried scutellaria baicalensis, schisandra chinensis and red peony root, and the weight ratio of the raw licorice, the dried bupleurum chinense, the dried scutellaria baicalensis, the schisandra chinensis and the red peony root can be about 1:1:1:1:1, the Chinese herbal medicine extract is obtained by an extraction method comprising the following steps: mixing the raw licorice, the bupleurum chinense, the dried scutellaria baicalensis, the schisandra chinensis and the red paeony root in the prepared raw materials to form a raw material mixture, and then extracting the raw material mixture by using a solvent. Also in this particular embodiment, the solvent used to perform the extraction process may be ethanol, such as 30% ethanol, 75% ethanol, and the like.

In the above-mentioned application of the Chinese herbal medicine extract of the present invention for preparing a medicament for treating skin diseases, the prepared medicament for treating skin diseases may be in a form of topical administration or systemic administration, but is not limited thereto. In one embodiment, the prepared medicament for treating skin diseases is in a topical administration form, and examples of the topical administration form may include, but are not limited to, ointments, creams, liquids, gels, and the like.

The medicine for treating skin diseases prepared from any one of the Chinese herbal medicine extracts has high safety and does not produce adverse effects on individuals. The medicine for treating skin diseases prepared from any one of the Chinese herbal medicine extracts has excellent efficacy for treating various skin diseases, and particularly has excellent inflammation relieving effect for skin diseases with inflammation symptoms.

In another embodiment, for the use of the above-mentioned Chinese herbal medicine extract of the present invention for preparing a medicament for treating skin diseases, the use of any one of the above-mentioned Chinese herbal medicine extracts together with steroids for preparing a medicament for treating skin diseases may be further included. In this embodiment, the weight ratio of herbal extract to steroid may be, but is not limited to, about 5:1 to about 200: 1. In a particular embodiment, the weight ratio of herbal extract to steroid may be about 83: 1. In yet another specific embodiment, the weight ratio of herbal extract to steroid may be about 5: 1. In yet another specific embodiment, the weight ratio of herbal extract to steroid may be about 100: 1.

The steroid may be any steroid suitable for skin treatment, and may be, for example, dexamethasone (dexamethasone), fluorometholone (fluorometholone), medrysone (medrysone), betamethasone (betamethasone), triamcinolone (triamcinolone), prednisone (prednisone), prednisolone (prednisone), hydrocortisone (hydrocortisone), etc., but is not limited thereto.

In the above embodiment of the use of any one of the above Chinese herbal extracts and a steroid for the preparation of a medicament for the treatment of a skin disorder, the prepared medicament for the treatment of a skin disorder may be a topical administration form or a systemic administration form, but is not limited thereto. In a specific embodiment, the prepared medicament for treating skin diseases is in a topical administration form, and examples of the topical administration form may include, but are not limited to, ointments, creams, liquids, gels, and the like.

The medicine for treating skin diseases, which is prepared by any one of the Chinese herbal medicine extracts and the steroid, has excellent effects on treating various skin diseases, and particularly has excellent inflammation relieving effect on skin diseases with inflammation symptoms. In addition, the traditional Chinese medicine extract can have the effect of reducing the side effect caused by the matched steroid, and/or the effective dose required by matching the traditional Chinese medicine extract with the steroid can be reduced compared with the effective dose required by singly using the steroid.

In another embodiment of the present invention, there is provided a use of a herbal extract for the preparation of a medicament for ameliorating side effects of steroid treatment of skin diseases.

The steroid may be any steroid suitable for use in skin treatment. Examples of steroids suitable for use in skin treatment may include, but are not limited to, dexamethasone, fluoromethalone, medrysone, betamethasone, triamcinolone, prednisone, prednisolone, hydrocortisone, and the like.

The skin disease may include autoimmune related skin diseases or infectious skin diseases. Examples of the above autoimmune-related skin diseases include, but are not limited to, psoriasis, allergic dermatitis, atopic dermatitis, etc.

The side effects of steroid treatment of skin diseases mean any adverse conditions or conditions that may be caused by steroid treatment of skin diseases, such as skin thinning and/or vasodilation, but are not limited thereto.

In addition, for the use of the Chinese herbal medicine extract of the present invention for preparing a medicament for improving side effects of steroid treatment of skin diseases, the Chinese herbal medicine extract described herein may be any one of the Chinese herbal medicine extracts mentioned in the related paragraphs of the use of the aforementioned Chinese herbal medicine extract for preparing a medicament for treatment of skin diseases, and thus, the details thereof will not be repeated herein.

In addition, in the above-mentioned use of the herbal extract of the present invention for preparing a drug for ameliorating side effects of steroid treatment of skin diseases, the prepared drug may be in a form of local administration or systemic administration, but is not limited thereto. In one embodiment, the drug is prepared in a topical dosage form, and examples of such topical dosage forms may include, but are not limited to, ointments, creams, liquids, gels, and the like.

In yet another embodiment of the present invention, a pharmaceutical composition for alleviating a skin condition is provided.

The pharmaceutical composition for alleviating skin disorders of the present invention may include, but is not limited to, herbal extracts and steroids.

The Chinese herbal medicine extract contained in the pharmaceutical composition for alleviating skin diseases of the present invention can be any one of the Chinese herbal medicine extracts mentioned in the related paragraphs of the use of the preceding Chinese herbal medicine extract for preparing a medicament for alleviating skin diseases, and therefore, the details thereof are not repeated herein.

The steroid contained in the pharmaceutical composition for alleviating skin diseases of the present invention may be any steroid suitable for skin treatment. Examples of steroids suitable for skin treatment may include, but are not limited to, dexamethasone, fluoromethalone, medrysone, betamethasone, triamcinolone, prednisone, prednisolone, hydrocortisone, and the like.

In the pharmaceutical composition for treating skin diseases of the present invention, the contents of the herbal medicine and the steroid are not particularly limited as long as the resulting pharmaceutical composition has the effect of treating skin diseases. The weight ratio of the herbal extract to the steroid may be, but is not limited to, about 5:1 to about 200: 1. In one embodiment, the weight ratio of herbal extract to steroid may be about 83: 1. In another embodiment, the weight ratio of herbal extract to steroid may be about 5: 1. In yet another embodiment, the weight ratio of herbal extract to steroid may be about 100: 1.

The pharmaceutical composition for alleviating skin diseases according to the present invention may be in a form of topical administration or systemic administration, but is not limited thereto. In one embodiment, the pharmaceutical composition for alleviating skin diseases of the present invention, and examples of the topical dosage form include, but are not limited to, ointment, cream, liquid, gel, etc.

In another embodiment, the pharmaceutical composition for treating skin diseases of the present invention may further comprise a pharmaceutically acceptable vehicle, carrier or salt.

Pharmaceutically acceptable vehicles may serve as diluents, dispersants or carriers for the active ingredients. Pharmaceutically acceptable vehicles can include materials commonly used in skin care products such as water, liquid or solid emollients, silicone oils, emulsifiers, solvents, humectants, thickeners, powders, propellants and the like.

The vehicle may constitute 80-99.9 wt%, preferably 85-95 wt% of the above composition and may constitute the remainder of the composition in the absence of other adjuvants.

Also, the compositions may further comprise other specific skin benefit activators such as sunscreens and skin lightening agents. Vehicles may also further include adjuvants such as antioxidants, fragrances, opacifiers, preservatives, colorants, and buffers.

In addition, all of the compositions described above can be made into a skin paint in one embodiment, including, but not limited to, an emulsion, a cream, a gel, a spray, a lotion, a shampoo, a mousse, and the like. Generally, skin sprays can be composed of spray copolymers, such as polyvinylpyrrolidone, vinyl acetate, and the like, and can function as lotions. Skin gels are prepared in a manner similar to sprays, but are gelatinous and free of the presence of alcohol and can be adhered to the skin. Skin curtains are foams released by pressure. The skin emulsion is a hydrophobic or hydrophilic emulsion, ointment, gel, emollient, spray, paint, skin conditioner, shampoo or mousse. In addition, suitable ingredients may be added to the skin cream, such additional ingredients including petrolatum, waxes, lanolin, silicones, liposomes, vegetables, mineral oils, plasticizers, fragrances, preservatives, penetration enhancers, pH adjusters or other ingredients suitable for topical skin application. This additional ingredient wets the skin, stabilizes the active compound, increases the aforementioned composition-to-skin contact, provides topical area concentration, and controls the release of the composition.

The pharmaceutically acceptable carriers can include, but are not limited to, solvents, dispersion media, coatings, antibacterial and antifungal agents and isotonic and absorption delaying agents, and the like, which are compatible with pharmaceutical administration. For different modes of administration, the pharmaceutical composition can be formulated into dosage forms (dosage form) using conventional methods.

Also, the pharmaceutically acceptable salts may include, but are not limited to, salts including inorganic cations, for example, alkali metal salts such as sodium, potassium or ammonium salts, alkaline earth metal salts such as magnesium, calcium salts, salts containing divalent or tetravalent cations such as zinc, aluminum or zirconium salts. In addition, they may be organic salts such as dicyclohexylamine salts and methyl-D-glucamine, amino acid salts such as arginine, lysine, histidine, glutamic acid amide.

The pharmaceutical compositions of the present invention may be administered non-orally, via inhalation spray (inhalation spray), or by way of an implanted reservoir (implanted reservoir). Non-oral administration may include brushing the affected area, subcutaneous (subcutaneous), intradermal (intracutaneous), intravenous (intravenous), intramuscular (intramuramulular), intraarticular (intraarticular), arterial (intraspecific), intrasynovial (intracavitary), intrasternal (intrasternal), subarachnoid (intramural), intralesional (intradivision) injection, and infusion techniques.

The topical composition applied in the form of ointment, cream, liquid, gel, etc., may be, but is not limited to, such.

Oral compositions may be in the form of, but are not limited to, tablets, capsules, emulsions (emulsions), aqueous suspensions (aqueous suspensions), dispersions (dispersions), and solutions.

The pharmaceutical composition for alleviating skin diseases of the present invention has excellent effects on alleviating various skin diseases, and particularly has excellent effects on alleviating and/or treating skin inflammation for skin diseases with inflammatory symptoms. Also, in the pharmaceutical composition, the Chinese medicinal extract may have an effect of reducing side effects caused by the collocated steroid, and/or the effective dose required by the collocation of the steroid with the Chinese medicinal extract may be reduced as compared to the effective dose required when the steroid is used alone.

Examples

A. Preparation of various test extracts

1. External ointment dosage form

(1) Extracting Scutellariae radix, radix bupleuri, Glycyrrhrizae radix, radix Paeoniae Rubra and fructus Schisandrae Bicoloris with 30% ethanol solution to obtain extract, Scutellariae radix extract, bupleuri radix extract, and Scutellariae radix and bupleuri radix extract

The following four different compound and single Chinese herbal medicine formulas (1) scutellaria baicalensis, bupleurum chinense, liquorice, paeonia lactiflora and schisandra chinensis compound medicinal materials (weight ratio is 1:1:1:1) (PT-A), (2) scutellaria baicalensis single formula (SR), (3) radix bupleuri single formula (BR), and (4) scutellaria baicalensis and radix bupleuri compound medicinal materials (weight ratio is 1:1) (SR + BR) are respectively subjected to the following mutextraction procedures:

the Chinese herbal medicine formula is heated and extracted for 1 hour by using 30% ethanol solution with the weight being 10 times that of the Chinese herbal medicine formula to obtain first extract liquid. The first extract was taken out and the extracted formulation was extracted again with 10 times the weight of 30% ethanol solution for another 1 hour to obtain a second extract. Then, the two extracts were mixed. The obtained extract was filtered through a 100mesh screen, concentrated under reduced pressure, and then freeze-dried.

The Chinese herbal medicine formulation is prepared by mixing lyophilized powder (1g) with 95% alcohol (1.4g), polyethylene glycol 400(10g), polyethylene glycol 4000(4.5g), and polyoxyethylene hydrogenated castor oil (0.15g), and high purity deionized water (2.95ml), heating to 60 deg.C, and mixing to even to make into topical ointment.

That is, the following four ointments were obtained: (1) the ointment comprises (by weight ratio) an ointment of a scutellaria baicalensis, a bupleurum chinense, liquorice, peony and a schisandra chinensis compound medicinal material (PT-A), (2) an ointment of a scutellaria baicalensis single prescription (SR), (3) an ointment of a bupleurum chinense single prescription (BR) and (4) an ointment of a scutellaria baicalensis and a bupleurum chinense compound medicinal material (SR + BR) in a ratio of 1: 1.

(2) Respectively extracting each single material with 30% ethanol solution, and mixing the extracts to obtain mixed extract

The following five kinds of Chinese herbal medicines, including baikal skullcap root, bupleurum root, licorice, peony and schisandra chinensis, are extracted separately:

a single Chinese herbal medicine is heated and extracted for 1 hour by using a 30% ethanol solution with the weight being 10 times that of the single Chinese herbal medicine to obtain a first extract liquid. The first extract was taken out and the extracted formulation was extracted again with 10 times the weight of 30% ethanol solution for another 1 hour to obtain a second extract. Then, the two extracts were mixed. The resulting extract was filtered through a 100mesh screen.

Namely, obtaining the filtrate of the single prescription of radix scutellariae, the single prescription of bupleurum chinense, the single prescription of liquorice, the single prescription of peony and the single prescription of schisandra chinensis respectively.

Mixing the above five filtrates to obtain mixed filtrate (PT-A-1) of Scutellariae radix, bupleuri radix, Glycyrrhrizae radix, radix Paeoniae and radix Schisandrae Bicoloris, concentrating under reduced pressure, and freeze drying.

Freeze-dried powder (1g), 95% alcohol (1.4g), polyethylene glycol 400(10g), polyethylene glycol 4000(4.5g), and polyoxyethylene hydrogenated castor oil (0.15g), and high purity deionized water (2.95ml) were heated to 60 deg.C and mixed to make ointment.

Namely, obtaining an ointment of an mutextract mixture (PT-A-1) of scutellaria baicalensis, bupleurum chinense, liquorice, paeonia lactiflora and schisandra chinensis mutextracted by 30% ethanol.

(3) Extracts of Scutellariae radix, bupleuri radix, Glycyrrhrizae radix, radix Paeoniae and radix Schisandrae Bicoloris compound medicinal materials extracted by 75% ethanol solution

Extracting Scutellariae radix, bupleuri radix, Glycyrrhrizae radix, radix Paeoniae and fructus Schisandrae Bicoloris compound with 10 times of 75% ethanol solution under heating for 1 hr to obtain first extractive solution. The first extract was taken out and the extracted formulation was extracted again with a 10-fold weight of 75% ethanol solution for another 1 hour to obtain a second extract. Then, the two extracts were mixed. The obtained extract was filtered through a 100mesh screen, concentrated under reduced pressure, and then freeze-dried.

The lyophilized powder (1g) was mixed with 95% ethanol (1.4g), polyethylene glycol 400(10g), polyethylene glycol 4000(4.5g), and polyoxyethylene hydrogenated castor oil (0.15g), and high purity deionized water (2.95ml) and heated to 60 deg.C and mixed well to make an ointment for external use.

Namely, obtaining the ointment of 75 percent ethanol mutextracted compound medicinal materials (weight ratio is 1:1:1:1:1) (PT-A-2) of scutellaria baicalensis, bupleurum chinense, liquorice, peony and schisandra chinensis.

(4) Steroid ointment

Betamethasone dipropionate powder (0.006g), 95% alcohol (1.4g), polyethylene glycol 400(10g), polyethylene glycol 4000(4.5g), polyoxyethylene hydrogenated castor oil (0.15g), and high-purity Deionized (DI) water (3.944ml) were heated to 60 ℃ and mixed uniformly to prepare an external ointment formulation (BD).

(5) Mixed ointment of compound medicinal materials of scutellaria baicalensis, bupleurum chinense, liquorice, paeonia lactiflora and schisandra chinensis and steroid

PT-A dry powder (0.5g), betamethasone dipropionate powder (0.006g), 95% alcohol (1.4g), polyethylene glycol 400(10g), polyethylene glycol 4000(4.5g), polyoxyethylene hydrogenated castor oil (0.15g) and high-purity deionized water (3.444ml) are heated to 60 ℃ and mixed uniformly to prepare an mut mutexternal ointment preparation (PT-A + BD).

2. Liquid dosage form

70mg of PT-A dry powder was formulated in a liquid dosage form with 7ml of solvent (10% DMSO with 90% glyceryl tricaprylate) at a concentration of 10 mg/ml.

B. Experimental methods and results

1. Example 1

Efficacy assessment of compound composition for reducing Imiquimod (IMQ) -induced psoriasis-like animal model skin disease

An imiquimod-induced psoriasis-like animal model was used to evaluate the effect of extracts on reducing skin disease. The implementation steps are as follows:

balb/c mice (6-8 weeks old) were first shaved and divided into a control group, and four mutexperimental groups, wherein the four mutexperimental groups were PT-A (100mg) ointment-treated group, SR (100mg) ointment-treated group, BR (100mg) ointment-treated group, and SR + BR (100mg) ointment-treated group.

None of the mice were given any treatment in the control group; in the control group, 62.5mg of imiquimod cream (Aldara; 3M Pharmaceuticals) was applied to the back of the mice, and the imiquimod cream was administered once a day for six consecutive days to induce psoriasis symptoms in the skin of the mice; in the four groups of experimental groups, 62.5mg of imiquimod cream was applied to the back of the mice, then the test ointment (100mg) was applied topically to each group, and the imiquimod cream and each experimental group ointment were administered once a day for six consecutive days.

The animals were then evaluated for the extent of skin disease on the back, including items such as redness, desquamation, and photographs. After the animals are sacrificed, the skin at the back inflammation part is taken to carry out the detection of the gene expression quantity of tissues, including IL-17A, IL-6, TNF-alpha, S100A7 and the like. The results are shown in FIGS. 1A and 1B.

Fig. 1A shows that the control group had a significant tendency to cause redness and desquamation of the skin of mice when imiquimod was administered, compared to the control group, whereas the mice had reduced skin disease when imiquimod and PT-a were administered simultaneously in the mutexperimental group. However, the other three experimental groups did not improve the skin disease phenomenon in mice (FIG. 1A).

Fig. 1B shows that there was a significant increase in the gene mutexpression levels of IL-17A, IL-6, TNF- α, and S100a7 in the control group compared to the control group, whereas there was a trend toward a decrease in the gene mutexpression levels in the tissues of PT-a ointment-coated animals, where there was a significant difference in the mutexpression levels of IL-6, TNF- α, and S100a7 compared to the control group (p <0.05, p < 0.001).

2. Example 2

Efficacy evaluation of compound composition extraction mode for reducing psoriasis-like animal model skin diseases induced by imiquimod

The main mut mut mutexperimental method was the same as that of mut mut mutexample 1 (no control group was set), and the mut mut mutexperimental groups were changed to three groups of PT-A (100mg) ointment treatment group, PT-A-1(100mg) ointment treatment group, and PT-A-2(100mg) ointment treatment group to evaluate the effect of the mut mut mutextraction method on the efficacy.

Each experimental group was given imiquimod cream and test ointment (100mg) once daily for six consecutive days.

The animals were then evaluated for the degree of skin disease on the back, including items of redness, desquamation, etc., and were given 0-8 points (8 points for the most severe and 0 points for the asymptomatic) according to the severity of desquamation plus redness and photographed. The results are shown in FIGS. 2A and 2B.

As can be seen from fig. 2A and 2B, compared to the redness and desquamation phenomenon after administration of imiquimod in the control group, the skin disease phenomenon in mice was reduced after concurrent administration of imiquimod and 100mg PT-a in the mut mut mutexperimental group (fig. 2A), and the improvement of desquamation condition was significantly different (fig. 2B), while the skin disease condition in mice could not be significantly improved in PT-a-1 and PT-a-2 groups, respectively.

3. Example 3

Evaluation of efficacy of intraperitoneal injection of PT-A-DS on psoriasis skin diseases of mice

The effect of PT-a of the invention on skin disease was assessed by an imiquimod-induced psoriasis-like animal model. The implementation steps are as follows:

balb/c mice (6-8 weeks old) were first shaved for their back hair and divided into control, control and experimental groups.

None of the mice were given any treatment in the control group; control group 62.5mg of imiquimod-containing cream (Aldara; 3M Pharmaceuticals) was applied to the back of mice, and imiquimod cream was administered once a day for six consecutive days to induce psoriasis symptoms in the skin of the mice; in the mut mutexperimental group, 62.5mg of imiquimod cream was applied to the back of the mice, followed by administration of PT-A-DS (100mg/kg) by intraperitoneal injection (in the solvent of 10% DMSO and 90% glyceryl tricaprylate (glycyl tricanoate)), and administration of imiquimod cream and PT-A-DS (100mg/kg) once a day for six consecutive days.

Then, the mice were sacrificed, and the inflamed skin was taken down and soaked in 4% formaldehyde (formaldehyde) for fixation, and then the samples were cut in length to a thickness of 6 μm and then stained with hematoxylin-eosin (hematoxylin and eosin) to evaluate the degree of skin diseases including epithelial thickness, parakeratosis or hyperplasia. The results are shown in FIG. 3.

FIG. 3 shows that the control group showed redness, desquamation and thickening of the epithelium after administration of imiquimod to the control group, whereas the mutexperimental group showed reduction of skin diseases after simultaneous administration of imiquimod and intraperitoneal injection of PT-A.

4. Example 4

Evaluation of efficacy of PT-A in reducing psoriasis skin disease in mice

In this mutexample, the effect of PT-a ointment on skin disease was evaluated by an imiquimod-induced psoriasis-like animal model. The implementation steps are as follows:

balb/c mice (6-8 weeks old) were first shaved for their back hair and divided into a control group, a control group and two mut mutexperimental groups, wherein the two mut mutexperimental groups were PT-A (10mg) ointment treated group and PT-A (100mg) ointment treated group.

None of the mice were given any treatment in the control group; in the control group, 62.5mg of imiquimod cream (Aldara; 3M Pharmaceuticals) was applied to the back of the mice, and the imiquimod cream was administered once a day for six consecutive days to induce psoriasis symptoms in the skin of the mice; in the mut mutexperimental group, 62.5mg of imiquimod cream was applied to the back of the mice, followed by topical application of PT-A (10mg and 100mg), and imiquimod cream and PT-A ointment were administered once a day for six consecutive days.

The animals were then evaluated for the extent of skin disease on the back, including items such as redness, desquamation, and the like. After animals are sacrificed, the skin at the inflammatory part of the back is taken to carry out the detection of the gene expression quantity of tissues, including IL-17A, IL-23, IL-22 and the like, and the skin at the inflammatory part is taken down and soaked in 4% formaldehyde for fixation, then the samples are longitudinally cut into 6 mu m thickness and then are respectively carried out with CD68, CD4, Neutrophil, Ki-67 and CD31 antibodies for carrying out immune tissue staining to evaluate the phenomena of immune cell infiltration, cell proliferation, angiogenesis and the like at the skin disease part. The results are shown in FIG. 4A, FIG. 4B and FIG. 4C.

FIG. 4 shows that the control group caused significant skin thickening in the back of mice when imiquimod was administered, compared to the control group, whereas the mutexperimental group reduced skin disease when imiquimod and 100mg PT-A were administered simultaneously.

On the gene mutexpression level, compared with the control group, IL-17, IL-23 and IL-22 in the control group are obviously increased, while the gene mutexpression level of the tissue of the PT-A ointment-coated animal is reduced; the mut mutexpression levels of IL-17, IL-23 and IL-22 were significantly different in the 100mg PT-a group compared to the control group, while the mut mutexpression levels of IL-22 were significantly different in the 100mg PT-a group (p <0.05, p <0.01) (fig. 4B).

In addition, the simultaneous administration of imiquimod and 100mg PT-A in the mutexperimental group reduced the mutexpression or number of mouse skin CD68, CD4, neutrophiles, Ki-67, and CD31 relative to the administration of imiquimod in the control group (FIG. 4C).

5. Example 5

PT-A for reducing effect of 1-fluoro-2,4-dinitrobenzene (1-fluoro-2,4-dinitrobenzene, DNFB) on inducing animal contact dermatitis

In this mutexample, the effect of PT-A on reducing contact skin disease was evaluated by using a model of 1-fluoro-2,4-dinitrobenzene induced contact dermatitis in animals. The implementation steps are as follows:

the C57BL/6 mice were divided into control, positive control and experimental groups for different treatments. The following tests were performed after five consecutive days of first providing 0.5% DNFB to each group of mice (drug-coated parts of control group, positive control group and experimental group, abdomen, ear and ear, respectively).

In the control group, 0.2% DNFB was applied to the abdomen of mice; in the positive control group, sunderron (0.5mg/g) was applied to the mouse ear; in the mutexperimental group, PT-A (50mg) ointment was applied to the ears of mice.

After 18 hours of each treatment, the animals were evaluated for the extent of contact hypersensitivity and, after sacrifice, CD4 immunohistological staining of the inflamed skin was performed. The results are shown in FIGS. 5A and 5B.

FIGS. 5A and 5B show that the skin thickening and the number of CD4 infiltrates in the mice can be reduced after the test group is administered PT-A, compared with the control group to which 1-fluoro-2,4-dinitrobenzene is administered, which is similar to the test result of the positive control group to which Rinderon is administered.

6. Example 6

Comparison of PT-A efficacy evaluation with commercial products for reduction of psoriasis skin disease in mice

In this mutexample, efficacy comparisons of PT-a to commercial products were evaluated by an imiquimod-induced psoriasis-like animal model. The implementation steps are as follows:

balb/c mice (6-8 weeks old) were first shaved for back hair and divided into a control group, and three mutexperimental groups, i.e., PT-A (100mg) ointment-treated group, Rinderon-V (0.06%, Taiwan salt wild, China) ointment-treated group, and Daivon mutex (0.005%, LEO Pharma) ointment-treated group.

None of the mice were given any treatment in the control group; in the control group, 62.5mg of imiquimod-containing cream (Aldara; 3M Pharmaceuticals) was applied to the back of the mice, and the imiquimod cream was administered once a day for six consecutive days to induce psoriasis symptoms in the skin of the mice; the experimental group applied 62.5mg of imiquimod cream to the back of the mice, followed by topical application of the test ointment, with the imiquimod cream and test ointment administered once a day for six consecutive days.

The animals were then evaluated for the extent of skin disease on the back, including items such as redness, desquamation, and photographs. After the animals are sacrificed, the skin at the back inflammation part is taken to carry out the detection of the gene expression quantity of tissues, including IL-17A, TNF-alpha, IL-6, S100A7 and the like. The results are shown in FIGS. 6A and 6B.

Fig. 6A shows that the control group caused significant redness and desquamation of the skin of mice after administration of imiquimod with respect to the control group, whereas the mice suffered from skin diseases when imiquimod and PT-a or Rinderon were administered simultaneously in the mutexperimental group.

Fig. 6B shows a significant increase in the gene mutexpression levels of IL-17A, TNF- α, IL-6 and S100a7 in the control group compared to the control group, whereas the gene mutexpression levels tended to decrease in the tissues of PT-a or ridtron ointment-applied animals, with significant differences in the mutexpression levels of IL-6, TNF- α and S100a7 compared to the control group (p <0.05, p <0.01, p < 0.001). The expression level of the Daivonex group in TNF-alpha and S100A7 is significantly different from that of the control group, but the skin desquamation phenomenon of the mice is not significantly improved.

7. Example 7

Evaluation of efficacy of PT-A in combination with steroid ointment for reduction of psoriasis skin disease in mice

The effect of PT-a plus steroid ointment on skin disease was assessed by an imiquimod-induced psoriasis-like animal model. The implementation steps are as follows:

balb/c mice (6-8 weeks old) were first shaved for back hair and divided into control, control and three mut mutexperimental groups, BD (steroid) ointment treated, BD + PT-A ointment treated (PT-A to BD weight ratio 83:1) and Daivobet (LEO Pharma) ointment treated groups. Each animal in each group was treated with 25mg ointment.

In the control group, 62.5mg of imiquimod cream (Aldara; 3M Pharmaceuticals) was applied to the backs of mice, and the imiquimod cream was administered once a day for six consecutive days to induce psoriasis symptoms in the mice; in the control group, 62.5mg of imiquimod cream was applied to the back of the mice, followed by topical application of an ointment base; experimental groups 62.5mg of imiquimod cream was applied to the back of mice followed by topical application of test ointment, with the imiquimod cream and test ointment being administered once daily for six consecutive days.

The animals were then evaluated for the degree of skin disease on the back, including items such as desquamation, redness, and the like, and given 0-8 points (8 points for the most severe and 0 point for the asymptomatic) according to the severity of desquamation plus redness and photographed. After sacrifice, the skin of the inflamed part of the animal is taken down and soaked in 4% formaldehyde for fixation, then the sample is longitudinally cut into 6 mu m thickness, and then hematoxylin-eosin staining is carried out to evaluate the degree of skin diseases, including epithelial thickness, parakeratosis or hyperplasia, and the immune tissue staining is respectively carried out by Ki-67 antibody to evaluate the cell proliferation phenomenon of the skin disease part. The results are shown in FIGS. 7A to 7D.

As shown in fig. 7A to 7D, mice showed significant redness, desquamation and thickening of the epithelium after administration of imiquimod in the control group (fig. 7A and 7C), whereas mice showed significant reduction of skin disease including desquamation and redness after administration of imiquimod and BD + PT-a in the mutexperimental group (fig. 7B), and showed significant differences compared to the control group (p <0.05, < p <0.01, < p <0.001), epithelial thickening reduction (fig. 7C) and Ki-67 mutexpression reduction (fig. 7D), and the mice showed no significant improvement of skin disease in the anti-BD and Daivobet mutexperimental groups.

Example 8

PT-A combined with the intermediate steroid ointment is compared with the efficacy of the potent steroid on reducing psoriasis skin diseases of mice

The effect of PT-a plus a mesosteroid ointment on skin disease was assessed by an imiquimod-induced psoriasis-like animal model. The implementation steps are as follows:

first, the back hair of C57BL/6 mice (6-8 weeks old) was shaved, and the mice were divided into a control group, and five mut mut mut mut mutexperimental groups, i.e., fluocinonide (0.05%; intermediate steroid) ointment treatment group, fluocinonide + PT-A (0.25%) ointment treatment group (PT-A to fluocinonide weight ratio of 5:1), fluocinonide + PT-A (5%) ointment treatment group (PT-A to fluocinonide weight ratio of 100:1), PT-A (5%) ointment treatment group, and clobetasol propionate (0.05%) ointment treatment group. Each animal in each group was treated with 25mg ointment.

In the control group, 62.5mg of imiquimod cream (Aldara; 3M Pharmaceuticals) was applied to the backs of mice, and the imiquimod cream was administered once a day for six consecutive days to induce psoriasis symptoms in the mice; in the control group, 62.5mg of imiquimod cream was applied to the back of the mice, followed by topical application of an ointment base; experimental groups 62.5mg of imiquimod cream was applied to the back of the mice followed by topical application of 25mg of test ointment, with the imiquimod cream and test ointment being administered once daily for six consecutive days.

The animals were then evaluated for the degree of skin disease on the back, including three items of desquamation, redness, and skin thickness, and were given 0-12 points (12 points for the most severe and 0 points for the asymptomatic) according to the respective severity of the above three items and photographed. After sacrifice, the skin of the inflamed part of the animal was removed and fixed by soaking in 4% formaldehyde, and then the sample was cut longitudinally to a thickness of 6 μm and stained with hematoxylin-eosin to evaluate the degree of skin diseases including epithelial thickness, parakeratosis or hyperplasia. The results are shown in FIG. 8A-1, FIG. 8A-2, FIG. 8B-1, FIG. 8B-2 and FIG. 8C.

As can be seen from fig. 8A-1 and 8A-2 and 8B-1 and 8B-2, the mutexperimental group administered fluocinolone acetonide + PT-a simultaneously reduced the skin disease phenomena including redness, desquamation and epithelial thickening in mice compared to the control group (fig. 8A-1 and 8A-2 and 8B-1 and 8B-2), and showed significant differences compared to the control group (p <0.01, p <0.001) (fig. 8C). Among them, fluocinolone acetonide + PT-A in the mut mutexperimental group improved the degree of improvement of skin diseases in mice compared to the fluocinolone acetonide-treated group, including reduction of redness, desquamation, and epithelial thickening phenomena (FIG. 8A-1 and FIG. 8A-2 and FIG. 8B-1 and FIG. 8B-2), especially fluocinolone + PT-A (0.25%) ointment-treated group, and showed significant difference (# # p <0.01) compared to the fluocinolone (FIG. 8C). The fluocinolone plus PT-A (5%) ointment treatment group showed a tendency to improve the degree of improvement, but did not show a significant difference (ns) compared with the fluocinolone group.

8. Example 9

Analysis of PT-A on Effect on mouse ear skin thickness

ICR mice (7-9 weeks old) were first divided into a control group (Vehicle) and three mutexperimental groups, PT-A (25 mg/g content) ointment treatment group, Clobetasol Cream (Clobetasol Cream, 0.25mg/g content of CLOBETASOL propionate, mixed Cream base) and Tufujing hydrophilic ointment (TOPSYM Cream, 0.25mg/g content of fluocinolone acetonide, mixed Cream base).

Applying cream base (containing no effective component) 10mg once per day to ear of control group; the test groups were applied to the ears once a day with 10mg of the test cream for fourteen consecutive days.

The thickness of the ear was measured before application and every 2 to 4 days after application, and the thickness of the ear before application was set to 100%, and the change in the thickness of the ear was observed. After sacrifice, the skin of the inflamed part of the animal was removed and fixed by soaking in 4% formaldehyde, and then the sample was cut longitudinally to a thickness of 6 μm and stained with hematoxylin-eosin to evaluate the degree of skin diseases including epithelial thickness, parakeratosis or hyperplasia. The results are shown in FIGS. 9A and 9B.

As can be seen from fig. 9A and 9B, the ear thickness variation of the PT-a mutexperimental group was consistent with that of the control group, and did not cause significant variation, whereas the ear thickness of the two groups of the coltsfoot cream and the tobutin hydrophilic ointment after 7 to 14 days of application was 71-79% and 80-81% of the thickness before application, respectively, and had a significant tendency to become thinner (p <0.05, p <0.01 compared with the control group).

9. Example 10

Analysis of the Effect of PT-A on reducing skin thickness caused by steroids

In this mutexample, the effect on the thickness of the ear skin was evaluated by applying PT-A ointment mixed with steroid continuously. The implementation steps are as follows:

ICR mice (7-9 weeks old) were first divided into control (vehicle), control (Clobeasol) and two mut mut mut mutexperimental groups, PT-A (25 mg/g content) ointment treated and PT-A (25 mg/g content) mixed with Clobetasol Cream (0.25 mg/g Clobetasol + PT-A) (100: 1 weight ratio of PT-A to Clobetasol).

Applying cream base (containing no effective component) 10mg once per day to ear of control group; control ear was applied once daily with 10mg of Kelisu Cream (CLOBETASOL Cream, CLOBETASOL Cream, 0.25mg/g CLOBETASOL propionate content after mixing with Cream base); in both groups, 10mg of the test ointment was applied to the ears once a day for 14 days.

The thickness of the ear was measured before application and every 2 to 4 days after application, and the thickness of the ear before application was set to 100%, and the change in the thickness of the ear was observed. After sacrifice, the skin of the inflamed part of the animal was removed and fixed by soaking in 4% formaldehyde, and then the sample was cut longitudinally to a thickness of 6 μm and stained with hematoxylin-eosin to evaluate the degree of skin diseases including epithelial thickness, parakeratosis or hyperplasia. The results are shown in FIGS. 10A and 10B.

As can be seen from fig. 10A and 10B, the thickness variation of the PT-a group ears of the mutexperimental group was consistent with that of the control group, and did not cause significant variation; the thickness of the ears of the test group was significantly reduced in the PT-a mixed coltsfoot cream group, and was 77-83% of the thickness before application after 7-14 days of application, which was similar to 71-79% of the control group (p <0.05, p <0.01 compared to the control group), whereas the PT-a mixed coltsfoot cream group was about 86% of the thickness before application after 3 days of application, and was significantly 73% thicker than the control group (p <0.05 compared to the control group).

Example 11

Evaluation and comparison of the Effect of PT-A on the adverse skin reactions with the commercial products

The C57BL/6 mice (6-8 weeks old) were first shaved of their back hair and divided into control, control and experimental groups.

Mice were not given any treatment in the control group; in the control group, 10mg of Rinderon ointment is smeared on the back of the mouse and is administered once a day for 60 days continuously; in the mutexperimental group, 10mg of PT-A ointment was applied to the back of the mice and administered once a day for 60 consecutive days.

The animals were then evaluated for the extent of adverse skin reactions on their backs, including items such as dryness, redness, desquamation, and the like, and photographed. After sacrificing the animals, the skin of the back without ointment (control group) or the skin with ointment (control group and experimental group) was stained with hematoxylin-eosin (hematoxylin-eosin) as shown in fig. 11A and 11B.

Fig. 11A shows that the control group caused significant drying and desquamation of the skin of mice after imiquimod administration compared to the control group, whereas the skin of mice maintained a normal appearance after PT-a60 days in the experimental group, which was the same as the control group.

Fig. 11B shows that the control group caused significantly thinning phenomenon of the epidermis of the mice compared to the control group, whereas the epidermis of the skin of the mice maintained normal thickness after administration of PT-a60 days in the experimental group, which was the same as the control group.

Example 12

The results of the human skin repeated irritation and allergy tests.

In this mutexample, a total of 50 subjects were tested by first applying PT-A (25 mg/g) ointment to a low sensitivity patch and then covering the posterior sub-scapular area of the subjects. After 24 hours, the patch was removed. The test was repeated every 2 days, 3 times in a week and the irritation test was performed for 3 consecutive weeks. The redness and edema of the skin were observed for 9 results before the 2-9 tests and before the allergy test.

After 10-14 days, 50 subjects were again applied with PT-A ointment (content 25mg/g) to the posterior sub-scapular area. After 24 and 48 hours, the skin redness and edema were evaluated, respectively, as an allergy test. The results are shown in FIG. 12.

The results in FIG. 12 show that PT-A does not cause skin irritation and allergic reactions in humans.

Example 13

Results of repeated irritation test of PT-A on skin of white rabbit

First, 2.0-3.0kg of hairs on the back of 3 New Zealand white rabbits were shaved off, and then PT-A (content: 25mg/g) ointment was applied to gauze of 1 inch × 1 inch size, and fixed and attached to the right area of the back with an artificial dressing for 24 hours, after which the gauze and the artificial dressing were removed, and the skin to be tested was washed with distilled water and applied for 17 days continuously. The control substance (distilled water for injection for the first 10 days and no sample ointment for the last 7 days) was also applied to gauze of the same size and fixed in the left area with an artificial dressing. After 24 hours, the gauze and the artificial dressing were removed, and the skin to be tested was washed with distilled water and applied for 17 days.

After the artificial dressing and the gauze are removed every day, the skin of the animal is observed whether to generate erythema and edema, and the observation time is continuously 17 days. The skin irritation index was calculated from the score obtained by scoring the skin response criteria table, and the skin irritation index was evaluated by the calculated skin irritation index, and the results are shown in Table 1.

TABLE 1 repeated irritation test results for PT-A (25 mg/g content) ointment-treated white rabbit skin

Primary Irritation Score (PIS)

Primary stimulation score-control score

Number 1001: 19-3 ═ 16

Number 1002: 0-0 ═ 0

Number 1003: 8-1 ═ 7

Skin Single irritation index PII

Skin single irritation factor PII primary irritation score/number of test days

Number 1001: 16/17 ═ 0.9 (mild irritation)

Number 1002: 0/17 is 0 (no stimulation)

Number 1003: 7-17 is 0.4 (no stimulation)

The results in Table 1 show that the PT-A ointment showed a mild irritant response in 1 animal, while the remaining 2 were non-irritating.

Although the present invention has been described with respect to the preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. The application of a Chinese herbal medicine ethanol extract in preparing a medicament for treating skin diseases is disclosed, wherein the Chinese herbal medicine ethanol extract is prepared from liquorice, radix bupleuri, scutellaria baicalensis, schisandra chinensis and Chinese herbaceous peony, and the weight ratio of the liquorice, the radix bupleuri, the scutellaria baicalensis, the schisandra chinensis and the Chinese herbaceous peony is 1-5: 1-5: 1-5: 1-5: 1-5, wherein the Chinese herbal medicine ethanol extract is a 30% ethanol extract, the skin disease is psoriasis or contact dermatitis, and the medicine for treating the skin disease is a local administration dosage form or a systemic administration dosage form.

2. The use of the ethanolic extract of chinese herbal medicine for the preparation of a medicament for the treatment of a skin disorder according to claim 1, wherein said ethanolic extract of chinese herbal medicine is obtained by an extraction process comprising the steps of:

(a) mixing the licorice, the bupleurum, the scutellaria, the schisandra and the peony in the preparation raw material to form a raw material mixture; and

(b) and (3) carrying out an extraction process on the raw material mixture by using ethanol.

3. The use of the ethanol extract of Chinese herbal medicines according to claim 1 for the preparation of a medicament for treating skin diseases, wherein the licorice comprises raw licorice or prepared licorice, the bupleurum comprises raw bupleurum chinense or bupleurum scorzonerifolium, the scutellaria comprises dried scutellaria baicalensis or scutellariae barbatae, the schisandra comprises schisandra chinensis or schisandra chinensis, and the paeonia comprises paeonia lactiflora or paeonia lactiflora.

4. The use of the ethanol extract of Chinese herbal medicines according to claim 1 for the preparation of a medicament for treating skin diseases, wherein the licorice is raw licorice, the bupleurum falcatum, the scutellaria baicalensis is kullcap, the schisandra chinensis is schisandra chinensis, and the paeonia lactiflora is red peony.

5. The use of the ethanol extract of Chinese herbal medicines according to claim 4 for preparing a medicament for treating skin diseases, wherein the weight ratio of the raw licorice, the bupleurum chinense, the scutellaria baicalensis, the schisandra chinensis and the red peony root in the raw materials for preparation is 1:1:1:1: 1.

6. the use of the ethanolic extract of chinese herbal medicine for the preparation of a medicament for the treatment of a skin disorder according to claim 5, wherein said ethanolic extract of chinese herbal medicine is obtained by an extraction process comprising the steps of:

mixing the raw licorice, the bupleurum chinense, the calcined scutellaria baicalensis, the schisandra chinensis and the red peony root in the preparation raw materials to form a raw material mixture; and

and (3) carrying out an extraction process on the raw material mixture by using ethanol.

7. Use of the ethanol extract of Chinese herbal medicines according to claim 1 for the preparation of a medicament for the treatment of skin diseases, wherein the topical administration form comprises an ointment, a cream, a liquid or a gel.

8. The use of the ethanolic extracts of chinese herbal medicine of claim 1 for the preparation of a medicament for the treatment of a skin disorder, further comprising the use of said ethanolic extracts of chinese herbal medicine together with a steroid for the preparation of a medicament for the treatment of a skin disorder.

9. Use of the ethanolic extracts of chinese herbal medicine according to claim 8, for the preparation of a medicament for the treatment of a skin disorder, wherein the weight ratio of said ethanolic extracts of chinese herbal medicine to said steroid is between 5:1 and 200: 1.

10. Use of ethanolic extracts of Chinese herbal medicines according to claim 8 for the preparation of a medicament for the treatment of skin diseases, wherein said steroid comprises dexamethasone, fluoromethalone, medrysone, betamethasone, triamcinolone, prednisone, prednisolone or hydrocortisone.

11. Use of the ethanolic extracts of Chinese herbal medicine according to claim 8 for the preparation of a medicament for the treatment of a skin disorder, wherein the topical administration form comprises an ointment, a cream, a liquor or a gel.

12. Use of a Chinese herbal medicine ethanol extract for preparing a medicament for improving the side effect of thinning skin caused by steroid treatment of skin diseases, wherein the Chinese herbal medicine ethanol extract is prepared from licorice, bupleurum, scutellaria, schisandra and peony in a weight ratio of 1-5: 1-5: 1-5: 1-5: 1-5, the Chinese herbal medicine ethanol extract is 30% ethanol extract, and the medicine for treating the skin diseases is a local administration dosage form or a systemic administration dosage form.

13. Use of the ethanolic extract of chinese herbal medicine according to claim 12 for the preparation of a medicament for ameliorating the side effects of skin thinning resulting from steroid treatment of skin disorders, wherein said ethanolic extract of chinese herbal medicine is obtained by an extraction process comprising the steps of:

14. The use of the ethanol extract of chinese herbal medicines according to claim 12 for the preparation of a medicament for ameliorating the side effects of skin thinning caused by steroid treatment of skin diseases, wherein the licorice comprises raw licorice or prepared licorice, the bupleurum comprises raw bupleurum chinense or bupleurum scorzonerifolium, the scutellaria baicalensis comprises scutellaria baicalensis or scutellaria barbata, the schisandra chinensis comprises schisandra chinensis or schisandra chinensis, and the paeonia lactiflora comprises paeonia lactiflora pall or paeonia lactiflora pall.

15. The use of the ethanolic extract of Chinese herbal medicine for the preparation of a medicament for ameliorating the side effects of skin thinning resulting from steroid treatment of skin disorders according to claim 12, wherein said licorice is raw licorice, said bupleurum falcatum is bupleurum falcatum, said scutellaria is dry scutellaria, said schisandra is schisandra chinensis and said peony is red peony.

16. The use of the ethanolic extract of Chinese herbal medicine according to claim 15, for the preparation of a medicament for ameliorating the side effects of thinning of skin caused by steroid treatment of skin disorders, wherein the weight ratio of the raw licorice root, the bupleurum chinense, the scutellaria baicalensis, the schisandra chinensis and the red peony root in the raw materials for the preparation is 1:1:1:1: 1.

17. use of the chinese herbal medicine ethanol extract according to claim 16 for the preparation of a medicament for ameliorating the side effects of skin thinning resulting from steroid treatment of skin diseases, wherein the chinese herbal medicine ethanol extract is obtained by an extraction method comprising the steps of:

18. Use of ethanolic extracts of Chinese medicinal herbs according to claim 12 for the preparation of a medicament for ameliorating the side effects of skin thinning resulting from treatment of skin disorders with steroids, wherein said steroids include dexamethasone, fluorometholone, medrysone, betamethasone, triamcinolone, prednisone, prednisolone, or hydrocortisone.

19. Use of the ethanolic extracts of Chinese herbal medicine according to claim 12 for the preparation of a medicament for ameliorating the side effects of skin thinning resulting from steroid treatment of skin disorders, wherein the topical administration form comprises an ointment, cream, liquid or gel.

20. A pharmaceutical composition for reducing skin disorders, comprising:

the Chinese herbal medicine ethanol extract is prepared from the following raw materials

Liquorice, radix bupleuri, scutellaria baicalensis, schisandra chinensis and Chinese herbaceous peony, wherein the weight ratio of the liquorice, the radix bupleuri, the scutellaria baicalensis, the schisandra chinensis and the Chinese herbaceous peony is 1-5: 1-5: 1-5: 1-5: 1-5; and

(ii) a steroid in a form selected from the group consisting of,

wherein the Chinese herbal medicine ethanolic extract has the efficacy of reducing the side effects of skin thinning caused by the steroid and/or the efficacy of reducing the effective dose of the steroid required to treat psoriasis or contact dermatitis compared to the steroid alone, the Chinese herbal medicine ethanolic extract is a 30% ethanolic extract, and the pharmaceutical composition is in a topical dosage form or a systemic dosage form.

21. The pharmaceutical composition for alleviating skin disorders of claim 20, wherein the weight ratio of the herbal ethanol extract to the steroid is 5:1 to 200: 1.

22. The pharmaceutical composition for alleviating skin disorders of claim 20 wherein the steroid comprises dexamethasone, fluoromethalone, medrysone, betamethasone, triamcinolone, prednisone, prednisolone, or hydrocortisone.

23. The pharmaceutical composition for alleviating skin disorders of claim 20, wherein the pharmaceutical composition is in a form for topical administration, wherein the form for topical administration comprises an ointment, cream, liquid, or gel.