Traditional Chinese medicine composition for treating eczema and preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine preparations, and provides a traditional Chinese medicine composition for treating eczema and a preparation method and application thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 1-3 parts of radix stemonae, 0.5-1.5 parts of rhododendron delavayi and 0.04-0.06 part of cantharides, and the specific preparation method comprises the following steps: respectively crushing the radix stemonae and the rhododendron delavayi into 20-100-mesh coarse powder, adding 500ml of 75% alcohol, soaking for 7 days, filtering, and complementing the filtrate to 500ml; and additionally taking cantharides, adding the filtrate into the cantharides, soaking for 10 days, filtering, filtering out liquid medicine, and sub-packaging into 50ml small bottles with the medicine concentration of 0.765 g.kg <-1 >, thereby obtaining the traditional Chinese medicine composition for treating eczema. The traditional Chinese medicine composition has different degrees of effects in each stage of wound repair of injured skin tissues, has no irritation or weak irritation of the allergy and irritation of the whole formula, is good in application safety and safe in clinical use, meets the standard requirements, and is worthy of further clinical popularization and application.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine preparation technology, specifically relating to a traditional Chinese medicine composition for treating eczema, its preparation method, and its application. Background Technology

[0002] Dermatitis is a general term for inflammatory skin conditions caused by various internal and external infectious or non-infectious factors. It is not a single disease. Its etiology and clinical manifestations are complex and diverse, and it is prone to recurrence, making clinical treatment difficult. Currently, most opinions consider dermatitis to refer to inflammation of the skin, such as contact sensitization and skin infections. The International Classification of Diseases (ICD-10) includes more than 20 diseases in the chapter on dermatitis and eczema, such as contact dermatitis, atopic dermatitis, and neurodermatitis. Clinically, a specific type of dermatitis is often defined based on its etiology, location, or other clinical characteristics. For example, dermatitis caused by contact with a certain substance is called contact dermatitis, and dermatitis caused by internal medications is called drug-induced dermatitis.

[0003] Atopic dermatitis, also known as atopic eczema, formerly called "atopic dermatitis" or "hereditary allergic dermatitis," is a chronic, relapsing, inflammatory skin disease characterized by significant dryness and itching. Because patients often have co-existing atopic diseases such as allergic rhinitis and asthma, it is considered a systemic disease. Atopic dermatitis is currently incurable and can severely impact quality of life. However, effective and standardized treatment can alleviate symptoms and reduce recurrence. Its etiology is currently unclear, but it is related to genetic factors, environmental factors, and disruption of the skin barrier function. Disruption of the skin barrier function can increase skin permeability, leading to increased water loss, resulting in dry, flaky skin. Furthermore, allergens can easily enter the body through the skin, triggering or exacerbating immune responses. In the early stages, atopic dermatitis is easily confused with other skin diseases, leading to misdiagnosis or delayed treatment. Recurrence is difficult to cure, and some patients, in pursuit of rapid results, incorrectly use steroid medications, causing side effects such as further damage to the skin barrier.

[0004] Limited treatment options: Although there are various treatment options, such as topical application of traditional Chinese medicine, antihistamines, hormone ointments, immunomodulators, and phototherapy, these methods may not meet the needs of all patients and have certain side effects and risks.

[0005] Currently available topical medications for treating atopic dermatitis include corticosteroids (TCS), calcineurin inhibitors (TCI), and AhR modulators (benvimod / Zelimi). While these medications demonstrate strong anti-inflammatory effects and symptom relief, long-term use may cause local adverse reactions such as skin atrophy, telangiectasia, and pigmentation. Prolonged, large-area use can even lead to hypothalamic-pituitary-adrenal axis suppression. Therefore, it is necessary to develop a drug that is effective, has few side effects, and is cost-effective. Summary of the Invention

[0006] This invention provides a traditional Chinese medicine composition for treating eczema, its preparation method, and its application.

[0007] This invention is achieved by the following technical solution: a traditional Chinese medicine composition for treating eczema, prepared from the following raw materials in parts by weight: 1-3 parts of Stemona japonica, 0.5-1.5 parts of Codonopsis pilosula, and 0.04-0.06 parts of Blisterella tamariscina. The specific preparation method is as follows: Stemona japonica and Codonopsis pilosula are pulverized into coarse powder of 20-100 mesh, added to 500ml of 75% alcohol, soaked for 7 days, filtered, and the filtrate is used to make up to 500ml; separately, Blisterella tamariscina is added to the above filtrate, soaked for 10 days, filtered, and the filtered liquid is dispensed into 50ml vials to obtain the final product; the drug concentration is 0.765g·kg. -1 .

[0008] Furthermore, the traditional Chinese medicine composition consists of 2 parts of Stemona japonica, 0.06 parts of Cantharides blister beetle, and 1 part of Codonopsis pilosula.

[0009] Furthermore, the traditional Chinese medicine composition is prepared into a tincture by adding commonly used excipients.

[0010] In this invention: Stemona japonica: sweet and bitter in taste, slightly warm in nature. It enters the Lung meridian. It moistens the lungs, lowers qi, stops cough, kills parasites and lice. It is used for acute and chronic cough, tuberculosis cough, and whooping cough; externally for head lice, body lice, pinworm infection, and vulvar itching.

[0011] *Codonopsis pilosula*: Bitter and sweet in taste; cool in nature; refers to the whole herb or root of *Codonopsis pilosula*, *Codonopsis lanceolata*, and *Codonopsis lanceolata* (all belonging to the Asteraceae family). It tonifies qi and benefits the lungs, relieves cough and asthma, clears heat and reduces fire (detoxifies). It is mainly used to treat bronchitis, tuberculosis, infantile malnutrition, and insufficient lactation. It also relieves cough and asthma, strengthens the spleen and aids digestion, and promotes lactation. It is primarily used for bronchitis, tuberculosis, infantile malnutrition, and insufficient lactation.

[0012] Cantharides: pungent in taste and hot in nature. It enters the liver, stomach, and kidney meridians. It breaks up blood stasis, disperses nodules and masses, and attacks toxins and erodes sores. It is used for abdominal masses, amenorrhea, stubborn tinea, scrofula, warts, carbuncles that do not ulcerate, and malignant sores with necrotic tissue.

[0013] The formulation principle of this invention is as follows: Stemona japonica, used externally to kill insects and lice, is the principal ingredient; Codonopsis pilosula, used to clear heat and detoxify, is the assistant ingredient; and Myxobolus praeruptorum, used to invigorate blood circulation, dissipate nodules, attack toxins, and erode sores, is the guiding ingredient. The three ingredients combined can clear heat, invigorate blood circulation, regulate menstruation, kill insects, relieve itching, and attack toxins. It is used to treat scabies, stubborn tinea, psoriasis vulgaris, eczema, and other diseases, with symptoms such as scaly erythema, swelling, blisters, itching, and skin ulceration.

[0014] Clinical application of this invention: From July 2023 to August 2025, 40 patients were treated, including 12 males and 28 females. The oldest patient was 73 years old, the youngest was 20 years old, and the average age was 39.7 years. The shortest duration of illness was more than 3 months, and the longest was more than 2 years. Patients applied the medication daily. One course of treatment lasted 15 days, and the efficacy was observed after one course.

[0015] Diagnostic basis: According to the provisions of the People's Republic of China Traditional Chinese Medicine Industry Standard - Standard for Diagnosis and Efficacy of Diseases and Syndromes in Traditional Chinese Medicine (ZY / T001.1-94), with reference to "Diagnostic basis for eczema". Acute eczema presents with polymorphic skin lesions, often exhibiting multiple forms simultaneously, including erythema, papules, vesicles, erosions, exudation, crusts, and desquamation. It has a rapid onset, with a burning sensation and intense itching. Lesions are often symmetrically distributed, most commonly on the head, face, distal extremities, and scrotum, and can spread throughout the body. It can develop into subacute or chronic eczema, with fluctuating severity and recurring relapses. Subacute eczema lesions have less exudation, mainly consisting of papules, papulovesicles, crusts, and scales, with mild erosions, a darker red color, and mild infiltration, accompanied by intense itching. Chronic eczema is mostly localized to a specific area, with clear borders, significant thickening and infiltration, a rough surface, or lichenification, and a brownish-red or brown color. It is often accompanied by papulovesicles, crusts, and scratch marks, tends to be moist, and frequently recurs, fluctuating in severity, with paroxysmal itching.

[0016] Syndrome Classification: Based on the provisions of the People's Republic of China Traditional Chinese Medicine Industry Standard—Standards for Diagnosis and Efficacy of Diseases and Syndromes in Traditional Chinese Medicine (ZY / T001.1-94), and referring to the "Syndrome Classification of Damp Sores". **Damp-Heat Infiltration:** Acute onset, with flushed and burning skin lesions, incessant itching, oozing, accompanied by fever, irritability, thirst, dry stools, scanty dark urine, red tongue, thin white or yellow coating, and slippery or rapid pulse; **Spleen Deficiency with Dampness Accumulation:** Slower onset, with flushed and itchy skin lesions, erosion and oozing after scratching, visible scales, accompanied by poor appetite, fatigue, abdominal distension, loose stools, pale and swollen tongue, white or greasy coating, and wiry and slow pulse; **Blood Deficiency with Wind-Dryness:** Long-term illness, with dark or discolored skin lesions, severe itching, or rough and thickened skin lesions, accompanied by dry mouth without thirst, poor appetite, abdominal distension, pale tongue, white coating, and thready and wiry pulse.

[0017] Efficacy evaluation: Based on the provisions of the People's Republic of China Traditional Chinese Medicine Industry Standard—Standards for Diagnosis and Efficacy of Diseases and Syndromes in Traditional Chinese Medicine (ZY / T001.1-94), and referring to the "Efficacy Evaluation of Eczema". Cured: Skin lesions subside; Improved: Skin lesions subside by more than 30%; Not cured: Skin lesions subside by more than 30% in a localized area.

[0018] Typical case: 1. Patient Zhou, male, 58 years old, with a history of chronic illness, presented with scattered erythema and papules on both lower limbs three years ago without any obvious cause. He experienced significant itching and treated himself with topical ointments, which improved the itching somewhat. However, the rash expanded further, affecting his entire body, including the trunk and limbs, with scattered erythema and papules. He was diagnosed with "eczema" and treated with oral doxycycline hydrochloride enteric-coated capsules, one tablet twice daily, and topical fluocinolone acetonide and vitamin B6 cream, but the effect was not satisfactory. He was then treated with oral ebastine tablets and topical mometasone furoate cream, which showed no improvement. The patient presented to our outpatient clinic seeking further integrated traditional Chinese and Western medicine treatment. Symptoms included: scattered edematous erythema on the scalp, trunk, and limbs, some of which merged into patches, particularly on the back; scattered papules ranging in size from rice grains to soybeans on the trunk and limbs, which showed significant scratching and crusting, with epidermal ulceration. During the course of the illness, the patient experienced no chest tightness, shortness of breath, abdominal pain, diarrhea, or other discomfort. Appetite was good, but sleep was poor, bowel movements were normal, tongue was pale red with a white coating, and pulse was deep. The patient was prescribed the topical medication described in Formula 2 of this invention, twice daily. After 15 days, the rash subsided, and itching was significantly relieved.

[0019] 2. Patient Chen, male, 63 years old, reported that two months prior, after sun exposure, he developed scattered millet-sized red papules on his head and hands. He self-medicated with an unknown topical medication, but the effect was poor. Gradually, the rash spread to his face and upper limbs, and he was diagnosed with "eczema." He was hospitalized and received intravenous compound glycyrrhizin injection and oral and topical medications (unknown). His symptoms did not improve significantly after one week. After discharge, he did not take the medication regularly and sought treatment at our outpatient clinic. His current symptoms included: scattered patchy erythema on the scalp, bright red in color; millet-sized to mung bean-sized erythema, papules, and nodules on the neck, upper limbs, back of hands, and buttocks; severe itching; visible scratch marks and scabs; good appetite; poor sleep; normal bowel movements; red tongue with a thin white coating; and a wiry and rapid pulse. He was given the topical medication described in Formula 2 of this invention, twice daily. After 10 days, the rash subsided, and the itching was significantly relieved.

[0020] 3. Patient Xu, male, 30 years old, presented with erythematous papules and itching on his buttocks and lower extremities more than two months prior, without any obvious cause. He sought medical attention at a community hospital and received intravenous fluids (details unknown), but the lesions did not improve. He then received oral and topical medications (details unknown), with limited effect. He subsequently presented to our outpatient clinic with the following symptoms: patchy erythematous papules on his buttocks and lower extremities, accompanied by severe itching. Mild swelling of both lower extremities, scattered scratch marks and crusts, good appetite, poor sleep due to itching, and normal bowel movements. His tongue was pale red with little coating, and his pulse was slippery and rapid. He was prescribed the topical medication described in Formula 2 of this invention, twice daily. After 21 days, the rash subsided, and the itching was significantly relieved.

[0021] Beneficial effects of this invention: This invention has different effects at various stages of wound repair of damaged skin tissue, and the allergenicity and irritation of the whole formula are non-irritating or weakly irritating. It has good application safety, is safe for clinical use, meets the standard requirements, and is worthy of further promotion and application in clinical practice. Attached Figure Description

[0022] Figure 1 This is a line graph showing the cumulative permeation of chlorogenic acid. Detailed Implementation

[0023] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are some embodiments of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains, and all materials publicly cited herein and cited by them are incorporated herein by reference.

[0025] Equivalent technologies of the specific embodiments described herein that are readily apparent to those skilled in the art through routine experimentation are included in this application.

[0026] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the instruments and equipment used in the following examples are all standard laboratory instruments and equipment; unless otherwise specified, the experimental materials used in the following examples were all purchased from regular biochemical reagent stores.

[0027] I. Experimental Materials 1. Main test reagents and instruments: 0.9% sodium chloride injection, Shijiazhuang Shiyao Co., Ltd., batch number: 2212311909, specification: 500ml; Rat IL-4 ELISA kit, Bio-Swamp Life Science Lab, batch number: 20230410; Rat TNF-α ELISA kit, Bio-Swamp Life Science Lab. Lab, Batch No.: 20230410; H1 Microplate Analyzer, Bertek (USA); TU-1901 Dual-Light UV-Vis Spectrophotometer, Beijing Purkinje General Instrument Co., Ltd.; KDC-2046 Low-Speed ​​Refrigerated Centrifuge, Keda Innovation Co., Ltd. Zhongjia Branch; H2050R Benchtop High-Speed ​​Refrigerated Centrifuge, Hunan Xiangyi Laboratory Instrument Development Co., Ltd.; Constant Temperature Water Bath, Tianjin Test Instrument Co., Ltd.; KP-XW80 Vortex Vortex Mixer, KEP Biotechnology Co., Ltd.; Hemorheology Analyzer, Sekhid 7000; Fully Automated Biochemical Analyzer, Shenzhen Raydu Life Science Co., Ltd.; BP-2000 Non-invasive Blood Pressure Analyzer, Visitech (USA); Eppendorf Micropipette, Eppendorf Research Plus, Germany; Clean Bench, Suzhou Purification Equipment Factory; LEICA RM2125 paraffin microtome, Leica Shanghai Branch; PM.10AD optical microscope, Olympus, Japan; Nikon optical microscope camera, Nikon Corporation, Japan.

[0028] 2. Experimental animals: Healthy KM rats, SFP grade, 160 in total, 80 males and 80 females, body weight 18 - 22 g, provided by Beijing Speyford Biotechnology Co., Ltd., license number: SCXK (Beijing) 2019 - 0010, batch numbers: 110324230101379042, 1103241911006681; Healthy SD breeding rats, SFP grade, 100 in total, 50 males and 50 females, body weight 180 g, provided by Beijing Speyford Biotechnology Co., Ltd., license number: SCXK (Beijing) 2022 - 0002, batch number: 111251230100080074; Healthy SD breeding rats, SFP grade, 80 in total, 40 males and 40 females, body weight 180 g, provided by Beijing Speyford Biotechnology Co., Ltd., license number: SCXK (Beijing) 2019 - 0010, batch number: 110324230102843927; DH guinea pigs, male, 80 in total, body weight 180 - 200 g, provided by Beijing Speyford Biotechnology Co., Ltd., license number: SCXK (Beijing) 2021 - 0008, batch number: 110329230100038035; New Zealand white rabbits, male, 24 in total, body weight (2.5 ± 0.2) Kg, provided by Beijing Changyang Xishan Farm, license number: SCXK (Beijing) 2021 - 0008, batch number: 110329230100038275. Clean and constant temperature environment, average temperature of the feeding environment: 25 °C, average humidity: 37%. Fresh air is supplied at 15 - 20 times per hour. Lighting: 12 hours of light and 12 hours of darkness. The basal diet is provided by Beijing Speyford Biotechnology Co., Ltd., license number: SCXK (Beijing) 2019 - 0010, batch number: 0328SH0412A. Animals can drink water and eat freely.

[0029] 3. Control drugs: Dexamethasone acetate tablets, 0.75 mg / tablet, produced by Tianjin Xinyi Jinjin Pharmaceutical Co., Ltd., approval number:国药准字H31020793, product batch number: 501211012. Amoxicillin and Clavulanate Potassium, calculated by amoxicillin as 250 mg / kg, produced by Lunan Bait Pharmaceutical Co., Ltd., approval number:国药准字H20053424, production batch number: 50220912.

[0030] 4. Modeling drugs: Xylene: produced by Tianjin Shentai Chemical Reagent Co., Ltd., used for the modeling of the experiment on the swelling of mouse ears induced by xylene with the traditional Chinese medicine composition described in this invention.

[0031] Formaldehyde: produced by Wuhan Sevier Biotechnology Co., Ltd., batch number: CR2209049, used for the modeling of the experiment on the swelling of mouse feet induced by formaldehyde with the traditional Chinese medicine composition described in this invention.

[0032] Staphylococcus aureus: Beijing Beina Chuanglian Biotechnology Research Institute - Microbial Quality Control Center, batch number: 220420; used for the replication of the rat bacterial dermatitis model.

[0033] II. Dosage design and selection of positive control drugs 1. Test drug: The formulation and drug concentration are as follows: Comparison of anti-inflammatory effects of different formulations: Prescription 1 (2 parts of Stemona japonica, 1 part of Codonopsis pilosula, 0.06 parts of Blisterella vesicularis), Prescription 2 (1.5 parts of Stemona japonica, 1 part of Codonopsis pilosula, 0.06 parts of Blisterella vesicularis), Prescription 3 (1 part of Stemona japonica, 1 part of Codonopsis pilosula, 0.06 parts of Blisterella vesicularis).

[0034] Drug grouping and concentration: Formula 1: Stemona japonica + Cantharides, 0.515 g / kg -1 Formula 2: Stemona japonica + Blisterella tataricus + Codonopsis pilosula, 0.765g / kg -1 Formula 5: Stemona japonica + Codonopsis pilosula, 0.75g / kg -1 Formula 6: Cantharides + Rejuvenating Ginseng, 0.265g / kg -1 The drugs used were extracted in the laboratory, and the specific method was as follows: Take Stemona japonica and Codonopsis pilosula and grind them into coarse powder of 20-100 mesh. Add 500ml of 75% alcohol and soak for 7 days. Filter and add the filtrate to make up to 500ml. Mix the drugs according to the groups. For Formula 1, Formula 2, and Formula 6, take Cantharides according to the proportions, add the corresponding filtrate, soak for 10 days, filter, filter out the liquid, and dispense into 50ml bottles to obtain the final product.

[0035] 2. Principles of Compound Dosage Design Mouse dosage: Positive group (dexamethasone acetate tablets, 2 mg / kg) -1 ), Fang Yi (Stemona japonica + Blisterella vesicatoria, 0.515g·kg) -1 Formula 2 (Stemona japonica + Blisterella vesicatoria + Codonopsis pilosula, 0.765g·kg) -1 ), Fang Wu (Bai Bu + Huan Yang Shen, 0.75g·kg) -1 Fang Liu (cantharides + Rejuvenating Ginseng, 0.265g·kg) -1 ).

[0036] Rat dosage: positive group (amoxicillin-clavulanate potassium, calculated as amoxicillin 250mg / kg, 0.114g / kg), formula 1 (stemona + blister beetle, 0.257g / kg), formula 2 (stemona + blister beetle + regenerating ginseng, 0.383g / kg), formula 5 (stemona + regenerating ginseng, 0.375g / kg), formula 6 (blister beetle + regenerating ginseng, 0.133g / kg).

[0037] Guinea pig dosage: Formula 1 (Stemona japonica + Blisterella vesicatoria, 0.257g / kg), Formula 2 (Stemona japonica + Blisterella vesicatoria + Codonopsis pilosula, 0.383g / kg), Formula 5 (Stemona japonica + Codonopsis pilosula, 0.375g / kg), Formula 6 (Blisterella vesicatoria + Codonopsis pilosula, 0.133g / kg). New Zealand rabbit dosage: 0.2ml (Stemona japonica + Cantharides blister beetle + Codonopsis pilosula, dosage is 0.408g·kg) -1 This conforms to the general practice of dose design in pharmacodynamic experiments of traditional Chinese medicine.

[0038] 3. Basis for selection of positive control drug and dosage design Dexamethasone acetate tablets: This product is already on the market and has a clear therapeutic effect in relieving acute inflammation. It serves as a reference drug for comparing the functions of Huanyang Shen compound tablets. The clinical dosage of dexamethasone acetate is: oral administration, the initial adult dose is 0.75-3 mg per dose, 2-4 times daily; specification: 0.75 mg per tablet. The equivalent mouse dose is 2 mg / kg. -1 .

[0039] Amoxicillin-clavulanate potassium: This product is suitable for various infections caused by susceptible bacteria. It is used as a Western medicine control in acute inflammation tests. The equivalent rat dose is 0.114 g / kg. -1 .

[0040] 4. Basis for Experimental Index Design Swelling degree and swelling inhibition rate: Swelling degree = mass of right earpiece - mass of left earpiece; Swelling inhibition rate (%) = (average swelling degree of blank control group - average swelling degree of treatment group) / average swelling degree of blank control group × 100%.

[0041] Serum marker detection: The experimental study investigated the effect of Huanyangshen compound on bacterial infectious dermatitis in rats, so it was necessary to detect inflammatory factors. Blood was collected from the abdominal aorta, and after the blood collection, the serum was separated and the serum IL-4 and TNF-α levels were detected by ELISA.

[0042] Pathological morphological observation of skin tissue: The effects of Huanyangshen compound on the pathological morphology of a rat model of bacterial infectious dermatitis were observed, mainly by detecting skin tissue.

[0043] Severity of skin allergic reaction: The animals were scored according to the "Scoring Criteria for the Severity of Skin Allergic Reaction," and the average score for each group was calculated. Simultaneously, the animals were observed for severe systemic allergic reactions such as asthma, unsteady gait, or shock. Sensitization was inferred based on the incidence rate of sensitization according to the "Evaluation Criteria for Skin Sensitization."

[0044] Severity of skin irritation reaction: Observe the erythema and edema of the animals according to the "Skin Irritation Reaction Scoring Standard" and score them. Observe whether there is pigmentation, bleeding points, rough or thin skin at the drug administration site and control site according to the "Skin Irritation Intensity Evaluation Standard". Determine the severity of irritation reaction based on the above scores.

[0045] III. Test Methods 1. Experimental study on xylene-induced ear swelling in mice: Model replication: Healthy KM mice, SPF grade, half male and half female, 18-22g. One hour after the last administration, 50μL of xylene, an inflammatory agent, was evenly applied to the right ear of each mouse. After 1 hour of inflammation, the mice were euthanized by cervical dislocation, and both ears were cut off. Ear pieces were punched at the same location in both ears using an 8mm diameter punch. The weight of each ear piece was measured, and the degree of ear swelling and the swelling inhibition rate were calculated.

[0046] Detection indicators: Swelling degree = Right earpiece mass - Left earpiece mass; Swelling inhibition rate (%) = (Average swelling degree of blank control group - Average swelling degree of drug treatment group) / Average swelling degree of blank control group × 100%.

[0047] Anti-inflammatory effect experimental study: Grouping and administration: Mice were randomly divided into blank group, model group, and positive group (dexamethasone acetate tablets, 2 mg / kg) according to body weight. -1 ), Fang Yi (Stemona japonica + Blisterella vesicatoria, 0.515g·kg) -1 Formula 2 (Stemona japonica + Blisterella vesicatoria + Codonopsis pilosula, 0.765g·kg) -1 ), Fang Wu (Bai Bu + Huan Yang Shen, 0.75g·kg) -1 Fang Liu (cantharides + Rejuvenating Ginseng, 0.265g·kg) -1 Each group consisted of 10 animals, and the corresponding medication was applied once daily for 3 consecutive days. The control group and the model group were given the same volume of physiological saline.

[0048] 2. Experimental study on formaldehyde-induced paw edema in mice Model replication: Healthy KM mice, SPF grade, half male and half female, 18-22g. After the last administration, 20μL of 10% formaldehyde was injected subcutaneously into the right hind paw pad of each mouse. After 1 hour of inflammation, the animals were sacrificed by cervical dislocation. The left and right hind paws were cut off at the same location on the hind paws, weighed, and the degree of paw swelling and swelling inhibition rate were calculated.

[0049] Detection indicators: Swelling degree = Right foot mass - Left foot mass; Swelling inhibition rate (%) = (Average swelling degree of blank control group - Average swelling degree of drug treatment group) / Average swelling degree of blank control group × 100%.

[0050] Formaldehyde-induced paw edema in mice: Grouping and administration: Mice were randomly divided into a blank group, a model group, and a positive group (dexamethasone acetate tablets, 2 mg / kg) according to body weight. -1 ), Fang Yi (Stemona japonica + Blisterella vesicatoria, 0.515g·kg) -1 Formula 2 (Stemona japonica + Blisterella vesicatoria + Codonopsis pilosula, 0.765g·kg) -1 ), Fang Wu (Bai Bu + Huan Yang Shen, 0.75g·kg) -1 Fang Liu (cantharides + Rejuvenating Ginseng, 0.265g·kg) -1Each group consisted of 10 animals, and the corresponding medication was applied once daily for 3 consecutive days. The control group and the model group were given the same volume of physiological saline.

[0051] 3. Experimental study on the effects of bacterial infectious dermatitis in rats Model replication: Healthy SD rats, SPF grade, half male and half female, 180-220g. Skin on the back of the rats was shaved (divided into A and B areas). A 20g weight was heated until red-hot, cooled for 10 seconds, and then applied to the rat's back, causing a burn for 10 seconds. The burn area was approximately 4cm × 4cm. The rats were then abraded with sandpaper (to the point of oozing blood). 1mL of Staphylococcus aureus (concentration 1×10 CFU / mL, solvent: physiological saline) was injected and allowed to distribute evenly naturally. The rats were then abraded to create a bacterial dermatitis model. A successful model was indicated by the appearance of copious amounts of yellow and white pus, ulceration, and redness at the affected area.

[0052] Grouping and drug administration: Rats were randomly divided into four groups according to body weight: blank group, model group, matrix group (75% ethanol), positive control group (amoxicillin-clavulanate potassium, 250 mg / kg amoxicillin, 0.114 g / kg), formula 1 (stemona + blister beetle, 0.257 g / kg), formula 2 (stemona + blister beetle + resurrection ginseng, 0.383 g / kg), formula 5 (stemona + resurrection ginseng, 0.375 g / kg), and formula 6 (blister beetle + resurrection ginseng, 0.133 g / kg), with 10 rats in each group. Twenty-four days after successful model establishment, the rats in each group were applied the corresponding drugs, while the rats in the blank group and model group were applied an equal volume of physiological saline. This was done once daily for 7 consecutive days.

[0053] General condition observation: Record and observe the skin healing rate of rats in each group on days 1, 3, 5 and 7 after administration, and record the scab formation, scab removal and healing of the wound (observe the general signs and manifestations of pus accumulation, redness, swelling, ulceration, etc. of rats in each group on days 1, 3, 5 and 7 after administration, and observe the scab formation, scab removal and healing of the wound with the naked eye).

[0054] Detection indicators: 24 hours after the last administration, anesthetized rats were given blood from the abdominal aorta. After blood collection, serum was separated and the levels of serum IL-4 and TNF-α were detected by ELISA.

[0055] Pathological observation: Rats were sacrificed, skin tissue was excised, fixed with 4% paraformaldehyde, stained with hematoxylin and eosin (HE), and the pathological changes of the skin tissue were observed under a microscope.

[0056] 5. Dermal toxicity test A. Acute Dermal Toxicity Test: SD rats, SPF grade, 180g-220g. Half male and half female. Adapted to laboratory conditions for 7 days. 24 hours before the test, the fur on both sides of the spine on the back of the animals was removed by pet grooming (approximately 4×4cm, divided into A and B areas). The skin was carefully examined, ensuring its integrity, and the rats were housed separately. Rats were randomly divided according to body weight into a blank group, a matrix group (75% ethanol), a positive control group (amoxicillin-clavulanate potassium, calculated as amoxicillin 250mg / kg, 0.114g / kg), Formula 1 (Stemona japonica + blister beetle, 0.257g / kg), Formula 2 (Stemona japonica + blister beetle + Codonopsis pilosula, 0.383g / kg), Formula 5 (Stemona japonica + Codonopsis pilosula, 0.375g / kg), and Formula 6 (blister beetle + Codonopsis pilosula, 0.133g / kg), with 10 rats in each group. The test sample was evenly applied to the hairless area A (right) of the experimental animals in the treatment group, and physiological saline was evenly applied to the hairless area B (left) of the experimental animals. Both sides were covered with oil paper and two layers of gauze, and then fixed with non-irritating adhesive tape or bandages to ensure close contact between the test sample and the skin, preventing it from falling off and the animals from licking the test sample. The test sample residue on the skin was washed off with warm water or a suitable solvent, and the local skin lesions and mortality were observed. The above operation was repeated after 4 hours and 8 hours. The toxicity of each rat was observed at 1 hour, 24 hours, 48 ​​hours, 72 hours and up to day 14 after drug administration, and detailed records were made.

[0057] B. Guinea pig skin allergy test Animals and Grouping: Fifty-six male guinea pigs were used. Before administration, the fur on both sides of their backs was shaved, creating a 4×4cm area (divided into A and B regions). They were randomly divided into a blank control group (distilled water, 0.2ml), a positive control group (1% 2,4-dinitrochlorobenzene, 0.2ml), a matrix group (75% ethanol), and six formulas: Formula 1 (Stemona japonica + blister beetle, 0.257g / kg), Formula 2 (Stemona japonica + blister beetle + Panax quinquefolius, 0.383g / kg), Formula 5 (Stemona japonica + Panax quinquefolius, 0.375g / kg), and Formula 6 (blister beetle + Panax quinquefolius, 0.133g / kg). Eight guinea pigs were housed in separate cages.

[0058] Sensitization exposure: The extract was evenly applied to the shaved area on side A. After application, the animals were separated into different cages and the treatment was repeated once at 7 days and 14 days, for a total of 3 times. For the positive group, 0.2 ml of 1% 2,4-dinitrochlorobenzene was evenly applied to the shaved area, and for the control group, 0.2 ml of distilled water was applied.

[0059] Challenge exposure: 14 days after the last administration, the extract was applied to the shaved area on the B side of guinea pigs. For the positive group, 0.2 ml of 0.1% 2,4-dinitrochlorobenzene was applied and left for 6 hours. The drug was then removed with warm water, and the skin allergic reaction was observed for 72 hours. Scoring was performed according to the "Scoring Criteria for the Severity of Skin Allergic Reactions" (Table 1), and the average score for each group was calculated. Simultaneously, animals were observed for severe systemic allergic reactions such as asthma, unsteady gait, or shock. Sensitization was inferred based on the incidence rate according to the "Evaluation Criteria for Skin Sensitization" (Table 2). Average reaction value = total score for erythema formation + total score for edema formation / total number of animals; sensitization rate = number of animals with erythema or edema ÷ total number of test animals.

[0060] Table 1: Scoring criteria for the severity of skin allergic reactions in guinea pigs Table 2: Evaluation Criteria for Skin Sensitization in Guinea Pigs 6. Rabbit skin local irritation test Animals and grouping: Healthy New Zealand white rabbits, approximately 2.5 kg, male, were compared using the whole-body left-right self-comparison method according to the requirements of the "Guidelines for Research on New Traditional Chinese Medicine".

[0061] Irritation test on intact skin: Hair was shaved on both sides of the back, with a 5×8cm area removed. The extract was applied once to side A, with a volume of approximately 0.2ml (dosage of 0.408g·kg). -1 Side B was evenly coated with 0.2 ml of distilled water as a blank control. 24 hours after hair removal, the hair-removed area was disinfected with 75% ethanol. Gauze soaked in the herbal medicine solution was then placed on the left back of the corresponding animal, covered with plastic wrap, and secured with non-irritating adhesive tape. The same method was used on the right side. After 24 hours, the medicine was washed off with damp gauze. Animals were then housed separately after medication application. At 1, 24, 48, 72 hours, and on day 7, the following data were recorded for each animal: weight, urination and defecation, respiration, diet, signs of systemic poisoning, and presence of erythema or edema at the application site. The average score for each group was calculated.

[0062] Preparation of damaged skin: Before administration, disinfect the hairless skin. Using a disposable sterile syringe needle, make a crisscross incision in the entire hairless skin, with the wound length 4-5 cm, allowing for oozing blood. The degree of damage on both sides of the skin should be basically the same. The administration method and observation indicators are the same as for intact skin. Each animal is housed separately, and administration is carried out continuously for 14 days. Skin reactions are observed and recorded daily. Observe and score the erythema and edema of the animals according to the "Skin Irritation Reaction Scoring Standard" (Table 3). Observe whether there is pigmentation, bleeding points, rough or thin skin at the administration site and control site according to the "Skin Irritation Intensity Evaluation Standard" (Table 4). Determine the degree of irritation reaction based on the above scores.

[0063] Table 3: Scoring Criteria for Skin Irritation Response in Rabbits Table 4: Evaluation Criteria for Skin Irritation Intensity in Rabbits 7. In vitro transdermal rate determination Chromatographic conditions: Determined according to high performance liquid chromatography (Chinese Pharmacopoeia, Part IV, General Chapter 0512, 2020) (operated in the dark).

[0064] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material; acetonitrile-0.4% phosphoric acid solution (13:87) was used as the mobile phase; the detection wavelength was 327 nm. The theoretical plate number, calculated based on the chlorogenic acid peak, should not be less than 12000.

[0065] Preparation of reference solution: Accurately weigh an appropriate amount of chlorogenic acid reference standard, add 50% methanol to prepare a solution containing 40 μg per ml.

[0066] Preparation of test solution: Filter the sample and collect the filtrate. Take 200 μl of the original sample solution for the transdermal diffusion test, filter it through a 0.22 μm filter membrane, and put it into a sample vial.

[0067] Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0068] Cumulative permeation and in vitro transdermal rate determination: Rat skin was prepared using rats. After routine anesthesia and cervical dislocation, the rats were shaved clean of hair on their backs with an electric razor. The skin was then dissected with surgical scissors, subcutaneous fat tissue was removed, and the skin was cut to appropriate size. After rinsing with physiological saline, the skin was used immediately. The isolated skin was divided into 1-hour and 24-hour administration groups, with three replicates in each group. In the 1-hour administration group, the surface moisture of the rat skin was blotted dry with lint-free paper. Using a Franz diffusion apparatus (TK-12D transdermal diffusion tester, manufacturer: Shanghai Kaikai Technology & Trade Co., Ltd.), the effective transdermal area was 3.14 cm². 2 The skin was fixed on a vertical diffusion chamber, with the dermis facing the receiving chamber (18 ml volume) and the stratum corneum facing the drug delivery chamber. The receiving chamber used 8 ml of 0.01% hydrochloric acid-methanol saline (3:7) as the receiving solution, rotated at 200 rpm, and incubated at 37°C for 30 min. Then, 2 ml of Formula II (Stemona japonica + Cantharides + Codonopsis pilosula) tincture was added to the drug delivery chamber. Samples of 500 μl were taken at 0.5, 1, 2, 4, 6, 8, and 12 h after the start of the experiment, and an equal volume of blank receiving solution at the same temperature was added simultaneously, with air bubbles removed. The sample solutions at each time point were injected and analyzed to calculate the chlorogenic acid content.

[0069] 8. Data Processing: Experimental data are expressed as mean ± standard deviation. It means that the following is adopted. SPSS 22.0 Statistical software was used for data processing, and analysis of variance was employed for measurement data. LSD-t For count data, the chi-square test was used, and the F-test was used to analyze the differences in the orthogonal experiment results. P<0.05 was considered statistically significant, and P<0.01 was considered statistically significant.

[0070] IV. Experimental Results 1. Comparison of anti-inflammatory effects of different formulations: All formulations with different ratios reduced ear swelling in mice to varying degrees, with formulation 2 (1.5 parts of Stemona japonica, 1 part of Codonopsis pilosula, and 0.06 parts of Cantharides) showing the best effect, as detailed in Table 5. Subsequent experiments used the drug ratios recorded in formulation 2 for grouped administration.

[0071] Table 5: Ear swelling and swelling inhibition rate of mice with different ratios ( (n=10) 2. Benzene-induced mouse ear swelling experiment: Compared with the control group, all treatments showed varying degrees of reduction in swelling, with the positive control group and treatments 2 and 6 showing the best effects and statistically significant differences compared with the control group. P <0.05); there were no significant differences among the treatment groups. See Table 6 for details.

[0072] Table 6: Mouse ear swelling degree and swelling inhibition rate ( (n=10) Note: Compared with the control group, ##P<0.01, #P<0.05.

[0073] B. Formaldehyde-induced paw edema test in mice: The results are shown in Table 7. Compared with the control group, each prescription reduced the edema to varying degrees, and the positive group showed a statistically significant difference compared with the control group. P <0.01), Formula 2 significantly reduced swelling, showing a statistically significant difference compared to the control group ( P <0.05), with no significant difference among the treatment groups. Compared with the control group, the positive control group, formula 2, and formula 5 showed higher swelling inhibition rates, with significant differences. P <0.05).

[0074] Table 7: Mouse paw swelling degree and swelling inhibition rate ( (n=10) Note: Compared with the control group, ##P<0.01, #P<0.05.

[0075] 3. Effects of bacterial infectious dermatitis in rats A. Observation of skin tissue characteristics in rats: Observe the general signs and manifestations such as pus accumulation, redness, swelling and ulceration on the 1st, 3rd, 5th and 7th days after administration, and observe the scab formation, scab removal and healing of skin wounds in each group of rats.

[0076] Indicator detection: The results are shown in Table 8. Compared with the blank group, the serum and skin tissue IL-4 levels in the model group were significantly increased; compared with the model group, each treatment group showed varying degrees of decrease in IL-4 levels, and the positive group showed statistically significant differences compared with formulas 2 and 5. P <0.05) or ( P <0.01). Compared with the blank group, the serum and skin tissue TNF-α levels in the model group were significantly increased; compared with the model group, each treatment group showed varying degrees of decrease in TNF-α levels, and the positive group showed statistically significant differences compared with formulas 2 and 5. P <0.05) or ( P <0.01). Compared with other compound treatment groups, the levels of IL-4 and TNF-α in rats treated with Formula 2 were lower, indicating a better anti-inflammatory effect.

[0077] Table 8: Effects on IL-4 and TNF-α levels in rats ( (n=10) B. Pathological observation: In the control group, the epidermis of the skin tissue was intact, with a clear structure and visible keratinized layer; the collagen fibers in the dermis were neatly arranged, and skin appendages such as hair follicles and sebaceous glands were scattered; the subcutaneous tissue, located below the dermis, consisted of loose connective tissue, adipose tissue, and muscle layer. Compared with the control group, the model group showed extensive necrosis in the skin tissue; large areas of epidermal necrosis with nuclear fragmentation (black arrows); large areas of necrosis were visible from the dermis to the subcutaneous tissue, with numerous hair follicles necrotic and structurally absent, and no obvious new hair follicle formation was observed. In the skin tissue of rats treated with Formulas 1, 2, and 5, fibrosis and inflammatory cell infiltration were observed, mainly characterized by fibroblast proliferation with oval or oblong nuclei, a small number of spindle-shaped nuclei, and less cytoplasm. A small number of neutrophils and lymphocytes were observed infiltrating the interstitium. Formula 2 shows increased fibrous tissue proliferation and thickening of the epidermis, which can promote wound healing to some extent. It may also be due to mast cells synthesizing interleukin or VEGF after administration, thereby promoting angiogenesis and affecting fibroblasts, regulating collagen remodeling, and promoting lesion healing. Formulas 2 and 6 show increased capillaries, possibly due to stimulating the body's immune CD4+ T lymphocytes to promote the production of vascular endothelial growth factor, leading to endothelial migration, proliferation, and angiogenesis.

[0078] The healing process of skin injury can be divided into four stages: hemostasis, inflammation, proliferation, and remodeling. Immediately after vasoconstriction and hemostasis, inflammatory cells are generated at the wound site to eliminate pathogens and cellular debris. Subsequently, granulation tissue proliferates through the migration of keratinocytes and fibroblasts to the wound and angiogenesis. Therefore, Formula 2 has varying degrees of effectiveness in each stage of wound repair in damaged skin tissue, making it the optimal choice.

[0079] 4. Dermal toxicity test A. Acute skin toxicity test: The results showed that no toxic reactions such as erythema or edema appeared on the skin of any group from the time the drug was washed off until 14 days after the drug administration. The mental state, behavior, diet, respiration, eyes and mucous membranes, feces and secretions of the rats were all normal, no rats died, and there were no significant changes in the body weight of the rats in any group. The changes in body weight of the rats in each group are shown in Table 9.

[0080] Table 9: Acute toxicity test of intact skin in rats ( (n=10) B. Guinea Pig Skin Allergy Test: Guinea pig skin allergy test scores are shown in Table 10. During the test, Huanyang Shen compound had no significant effect on the physiological activities of most experimental guinea pigs. No abnormalities were found in the body hair, skin pigmentation, normal physiological behavior, spontaneous activity, spontaneous feeding and drinking, secretions from the mouth, eyes, nose, and ears, the condition and frequency of physiological excretions, movement, and resistance muscle strength of the participating guinea pigs. This indicates that Huanyang Shen compound had no significant effect on the nervous system of guinea pigs. The compound contains ethanol and Chinese herbal irritant components, which have a mild irritant effect on the intact bare skin of guinea pigs. Slight erythema was visible after administration, but the duration was short and the skin color returned to normal in a short time. No scratching or biting behavior was observed in the treated area, and no edema was observed in the treated area and surrounding skin. No allergic reactions were observed. This indicates that Huanyang Shen compound formulas 1 and 5 have low irritation and toxicity to the intact skin of guinea pigs, and the irritation only occurred in the early stage of administration. The skin color returned to normal during the recovery period, and there was no acute or long-term toxicity, indicating high safety.

[0081] Table 10: Guinea Pig Skin Allergy Reaction Scores C. Rabbit Skin Irritation Test: Rabbit skin irritation response scores are shown in Table 11, and the average skin irritation scores are shown in Table 12. The skin condition at the administration site was observed daily in New Zealand rabbits during the 3-day administration period and for 3 days after the administration. The results showed that each prescription in the Huanyang Shen compound showed mild or no irritation in the rabbit skin irritation test; prescription 5 showed no or occasional erythema and edema at the administration site and control site (physiological saline) in both intact and broken skin groups, and was determined to be non-irritating based on the skin irritation level evaluation. No abnormalities such as drug-induced bleeding, pigmentation, or rough skin were observed. Furthermore, other physiological phenomena (drinking, eating, excretion, appearance, weight) of the experimental animals were also normal. Mild irritation was observed in the other administration groups. According to the national standards for skin irritation testing, skin reaction scoring, and reaction intensity evaluation, prescription 5 showed no irritation to either intact or broken skin in New Zealand rabbits. Based on the requirements of optimal efficacy and minimal toxicity, Formula 2 is selected as the best prescription. Its application safety is good, its clinical use is relatively safe, it meets the standard requirements, and it is worthy of further promotion and application in clinical practice.

[0082] Table 11: Rabbit Skin Irritation Response Scores Table 12: Average Skin Irritation Score of Rabbits 5. Cumulative Permeation and In Vitro Transdermal Rate Determination: The cumulative permeation and in vitro drug permeation rate of Formula 2 (Stemona japonica + Cantharides blisterii + Codonopsis pilosula) tincture are shown in Table 13. The cumulative permeation line graph is shown below. Figure 1 0.5 hours after administration, the cumulative permeation of chlorogenic acid began to increase significantly, and the upward trend slowed down until 8 hours after administration.

[0083] Table 13: Cumulative in vitro permeation and in vitro drug penetration rate of the compound (chlorogenic acid) This invention was used in acute inflammation tests in mice, bacterial dermatitis tests in rats, acute toxicity tests in rats, skin irritation tests in rabbits, and skin allergy tests in guinea pigs. The results showed that the Huanyang Shen compound was most effective in the xylene-induced mouse ear swelling test, particularly in the positive group and in formulas 2 and 5. In the formaldehyde-induced mouse paw edema test, formula 5 significantly reduced swelling. In the effect of Huanyang Shen compound on bacterial infectious dermatitis in rats, formulas 2 and 5 showed statistically significant differences. Formulas 1, 2, and 5 all showed fibrosis and inflammatory cell infiltration in the rat skin tissue, mainly characterized by fibroblasts with oval or oblong nuclei, a small number with long spindle-shaped nuclei, and less cytoplasm. A small number of neutrophils and lymphocytes were observed infiltrating the interstitial space. Formula 2 showed more fibrosis and thickened epidermis, which may promote wound healing to some extent. This may also be due to mast cells synthesizing interleukin or VEGF after administration, thereby promoting angiogenesis and affecting fibroblasts, regulating collagen remodeling, and promoting lesion healing. Formulas 2 and 5 show an increase in capillaries, which may be due to the stimulation of the body's immune CD4+ T lymphocytes to promote the production of vascular endothelial growth factor, thereby causing vascular endothelial migration and proliferation and angiogenesis.

[0084] Acute skin toxicity tests showed that no toxic reactions such as erythema or edema occurred in any group of rats within 14 days after drug removal. The rats' mental state, behavior, diet, respiration, eyes and mucous membranes, feces, and secretions were all normal, with no rat deaths and no significant changes in body weight in any group. Formula 2 was non-irritating to rabbit skin and did not produce significant skin allergic reactions in guinea pigs.

[0085] In vitro transdermal absorption rate assay results showed that 0.5 h after administration of Formula 2, the cumulative permeation of chlorogenic acid began to increase significantly, and the increasing trend slowed down until 8 h after administration. After 12 h, the transdermal absorption rate reached 12.52%.

[0086] In summary, Formula 2 has varying degrees of effect on wound repair of damaged skin tissue at all stages, and the formula is non-irritating or mildly irritating, with good application safety and clinical use, meeting the requirements of the standard, and is worthy of further promotion and application in clinical practice.

[0087] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine composition for treating eczema, characterized in that: This medicine is prepared from the following raw materials in parts by weight: 1-3 parts of Stemona japonica, 0.5-1.5 parts of Codonopsis pilosula, and 0.04-0.06 parts of Myxobolus thunbergii. The specific preparation method is as follows: Stemona japonica and Codonopsis pilosula are pulverized into coarse powder of 20-100 mesh, added to 500ml of 75% alcohol, soaked for 7 days, filtered, and the filtrate is used to make up to 500ml. Separately, Myxobolus thunbergii is added to the above filtrate, soaked for 10 days, filtered, and the liquid is dispensed into 50ml vials, with a drug concentration of 0.765g·kg⁻¹. -1 This refers to a traditional Chinese medicine composition for treating eczema.

2. The traditional Chinese medicine composition for treating eczema according to claim 1, characterized in that: The traditional Chinese medicine composition consists of 2 parts of Stemona japonica, 0.06 parts of Cantharides blister beetle, and 1 part of Codonopsis pilosula.

3. A traditional Chinese medicine composition for treating eczema according to claim 1 or 2, characterized in that: The traditional Chinese medicine composition is prepared into a tincture by adding commonly used excipients.

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