An antifungal skin disease agent containing radix paeoniae alba and radix sophorae flavescentis, and a preparation method and application thereof

Abstract

This invention proposes an antifungal dermatitis agent containing peony bark, white peony root bark, and sophora flavescens, its preparation method, and its application, belonging to the field of veterinary drug technology. It includes an active ingredient and other pharmaceutically acceptable excipients. The active ingredient comprises a traditional Chinese medicine extract and an antifungal compound in a mass ratio of (50-100):1. The traditional Chinese medicine extract is obtained by extracting peony bark, white peony root bark, and sophora flavescens in a mass ratio of (1-3):(2-5):(2-4). The antifungal compound has the structural formula shown in Formula I or a pharmaceutically acceptable salt thereof.

Description

Technical Field

[0001] This invention relates to the field of veterinary drug technology, specifically to an antifungal dermatological agent containing Danshen, Baishen, and Kushen, as well as its preparation method and application. Background Technology

[0002] Fungal skin diseases are superficial fungal infections, primarily caused by keratophilic dermatophytes, including Microsporum, Trichophyton, and Epidermophyton. These fungi are widespread globally, affecting various animals such as dogs and cats, and even humans. Healthy animals carrying the fungus gradually develop the infection under the influence of certain triggering factors. This process is likely closely related to the host's physiological condition and environmental factors. Triggering factors include malnutrition, changes in ambient temperature, and skin damage. Influenced by both environmental factors and individual immune differences, fungal infections spread rapidly, and infection rates remain high year-round.

[0003] Dermatophyte infection in dogs and cats is a common skin disease in small animals. In this disease, the normal flora on the body's surface is disrupted, leading to the over-adherence, proliferation, and invasion of pathogenic fungi into the skin tissue, causing a series of organic lesions. These fungi, which parasitize the fur, epidermis, and claws of dogs and cats, cause various skin diseases collectively known as dermatophyte infection. It is characterized by obvious circular patches of hair loss on the skin, accompanied by exudate, scaling, crusting, and itching. It is a zoonotic disease, medically termed "ringworm" in humans. Pet fungal dermatitis is relatively common in veterinary clinics and is difficult to cure, with a high recurrence rate.

[0004] Currently, chemical drugs are the primary method for treating fungal diseases. Clinically, drugs used to treat canine and feline dermatophytes mainly include broad-spectrum drugs such as antibiotics and corticosteroids. Although these drugs are effective against bacteria, fungi, and parasites, facilitating clinical treatment, long-term use of broad-spectrum drugs may lead to an imbalance in the animal's surface microecology, decreased immunity, and the development of drug resistance in microorganisms, potentially causing recurrent outbreaks. In recent years, with the emergence of drug-resistant strains and rising disease recurrence rates, researchers have actively sought novel, broad-spectrum, highly effective, and inexpensive antifungal drugs. Traditional Chinese medicine (TCM), as a traditional advantageous resource in my country, has gained widespread recognition for its advantages such as multiple targets, broad pathways of action, and low resistance to drug development. However, TCM alone has a slower onset of action and is less effective than Western medicine. Therefore, the combined use of TCM and Western medicine can significantly reduce the emergence of drug-resistant strains and achieve better therapeutic results. Summary of the Invention

[0005] The purpose of this invention is to propose an antifungal dermatological agent containing Danshen, Baishen, and Kushen, as well as its preparation method and application, to overcome the shortcomings of existing technologies such as strong drug resistance, large toxic side effects, and high recurrence rate, and to develop an agent that is resistant to drug resistance, has broad-spectrum antifungal activity, quickly relieves itching, promotes wound healing, is safe for external use, and is non-irritating.

[0006] The technical solution of this invention is implemented as follows:

[0007] This invention provides an antifungal dermatological agent containing peony bark, white peony root bark, and sophora flavescens, comprising an active ingredient and other pharmaceutically acceptable excipients. The active ingredient comprises a traditional Chinese medicine extract and an antifungal compound in a mass ratio of (50-100):1. The traditional Chinese medicine extract is obtained by extracting peony bark, white peony root bark, and sophora flavescens in a mass ratio of (1-3):(2-5):(2-4). The antifungal compound has the structural formula shown in Formula I or a pharmaceutically acceptable salt thereof.

[0008]

[0009] Formula I.

[0010] As a further improvement of the present invention, the preparation method of the traditional Chinese medicine extract is as follows:

[0011] S1. Weigh out the peony bark, dictamnus root bark, and sophora flavescens according to the proportion, pulverize them, pass them through a 40-mesh sieve to obtain a mixed powder, and divide it into two equal portions; the Chinese medicine components (paeonol, dictamnus root bark alkaloid, and matrine) have anti-inflammatory, antipruritic, and epidermal repair promoting effects.

[0012] Sophora flavescens: bitter and cold in nature, it clears heat and dries dampness, kills parasites and relieves itching. It mainly enters the lower burner and the skin, and is good at treating tinea, eczema and vulvar itching caused by damp-heat accumulation in the skin and insect infestation. It is the core ingredient in antifungal treatment.

[0013] Dictamnus dasycarpus root bark: Bitter and cold in nature, it clears heat and dries dampness, dispels wind, detoxifies, and relieves itching. It dispels wind-damp-heat toxins, treats scabies and urticaria, and assists Sophora flavescens in drying dampness and relieving itching, while also dispelling wind pathogens.

[0014] Peony bark: Bitter, pungent, and slightly cold in nature, it clears heat, cools the blood, and promotes blood circulation. Entering the blood, it clears heat from the skin, disperses blood stasis, reduces swelling, and relieves erythema and burning; it also prevents damp-heat from stagnating and causing blood stasis, and addresses itching caused by "blood heat generating wind".

[0015] Sophora flavescens kills parasites by drying dampness, Dictamnus dasycarpus dispels wind and dries dampness, and Paeonia suffruticosa cools the blood and disperses blood stasis. When used together, these three herbs dry dampness without harming Yin, dispel wind without depleting blood, and cool the blood without causing blood stasis, addressing both the root cause and the symptoms.

[0016] S2. Take 1 part of the mixed powder and add 8-12 times the mass of pure water, soak for 30-60 minutes, heat and reflux to extract for 1-2 hours, filter, add 6-10 times the mass of pure water to the filter residue, heat and reflux to extract for 0.5-1 hours, filter, combine the two filtrates, concentrate under reduced pressure, dry, and pulverize to obtain the water extract powder of traditional Chinese medicine.

[0017] S3. Take another portion of the mixed powder, add 8-10 times the mass of 40-60wt% ethanol solution, extract by ultrasonication for 10-30 min, filter, add 3-5 times the mass of 60-80wt% ethanol solution to the filter residue, heat and reflux for 1-2 h, filter, combine the two filtrates, recover the ethanol, concentrate under reduced pressure, dry, and pulverize to obtain the Chinese herbal alcohol extract powder.

[0018] S4. Mix the water extract powder and alcohol extract powder of traditional Chinese medicine evenly to obtain the traditional Chinese medicine extract.

[0019] As a further improvement of the present invention, the ultrasonic extraction power is 200-400W and the temperature is 40-60℃.

[0020] As a further improvement of the present invention, the method for preparing the antifungal compound is as follows:

[0021] T1. 2,4-bis(4-morpholino)-6-chloropyrimidine was reacted with m-aminophenylacetylene to prepare intermediate 1, with the following structure: ;

[0022] T2. Intermediate 1 was reacted with 2,4-dibromobenzyl azide to obtain the product.

[0023] As a further improvement of the present invention, the molar ratio of 2,4-bis(4-morpholino)-6-chloropyrimidine to m-aminophenylacetylene in step T1 is 1:1-1.2.

[0024] As a further improvement of the present invention, the molar ratio of intermediate 1 and 2,4-dibromobenzyl azide in step T2 is 1:1.2-1.5.

[0025] As a further improvement of the present invention, the dosage form of the agent is at least one of spray and ointment.

[0026] When the dosage form is a spray, the pharmaceutically acceptable excipients include: solvents, solubilizers, preservatives, humectants, and pH adjusters;

[0027] The composition (by weight percentage) of antifungal dermatology agents containing Danshen and Kushen is as follows:

[0028] 5-10 wt% Chinese herbal extract, 0.1 wt% antifungal compound, 20-40 wt% ethanol, 5-10 wt% propylene glycol, 1-3 wt% polysorbate-80, 0.1-0.2 wt% sodium benzoate, 2-5 wt% glycerin, and the remainder purified water.

[0029] When the dosage form is an ointment, the composition of the antifungal dermatitis agent containing Danshen, Baishen, and Kushen is as follows (by weight percentage): 7.5-15 wt% of Chinese herbal extract, 0.15 wt% of antifungal compound, 15-25 wt% of white petrolatum, 5-10 wt% of glyceryl monostearate, 10-15 wt% of liquid paraffin, 3-5 wt% of lanolin, 0.1-0.2 wt% of ethylparaben, and the balance of purified water.

[0030] This invention further protects a method for preparing the above-mentioned antifungal dermatitis agent containing Danshen and Sophora flavescens. The method involves mixing the Chinese herbal extract and the antifungal compound evenly to obtain the active component, adding excipients, and preparing the agent according to conventional formulation methods to obtain the antifungal dermatitis agent containing Danshen and Sophora flavescens.

[0031] This invention further protects the application of the above-mentioned antifungal dermatological agent containing Danshen and Sophora flavescens in the preparation of veterinary drugs.

[0032] This invention further protects the use of the above-mentioned antifungal dermatitis agent containing Danshen and Sophora flavescens in the preparation of veterinary drugs with antifungal effects on cats and dogs.

[0033] The present invention has the following beneficial effects:

[0034] 2,4-Bis(4-morpholino)-6-chloropyrimidine is an important organic synthesis intermediate. The pyrimidine nucleus is a DNA / RNA base analog that can interfere with fungal nucleic acid metabolism. The bismorpholino group enhances lipid solubility and improves cell membrane permeability. The chlorine atom at position 6 is substituted to introduce a triazole group, which is a good CYP51 inhibitor pharmacophore with good fungicidal and antifungal activity. The phenyl group attached to the triazole group further enhances lipid solubility and permeability. The antifungal compound prepared in this invention has a simple synthetic method, high yield, and high activity.

[0035] The active ingredients of this invention include the synergistic use of traditional Chinese medicine extracts and antifungal compounds, which greatly reduces drug resistance and has broad-spectrum antifungal activity. It also has the effects of rapid itch relief and promoting wound healing, and is safe for external use with no systemic toxicity or irritation. Attached Figure Description

[0036] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0037] Figure 1 The images show a comparison of scanning electron micrographs of severely infected areas in the control group, infection test group, and Example 3 group. A represents the control group; B represents the infection test group; and C represents Example 3 group. Detailed Implementation

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and 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.

[0039] Preparation Example 1: Traditional Chinese Medicine Extract

[0040] The preparation method is as follows:

[0041] S1. Weigh out peony bark, dictamnus root bark, and sophora flavescens in a mass ratio of 1:2:2, pulverize them, pass them through a 40-mesh sieve to obtain a mixed powder, and divide it into two equal portions.

[0042] S2. Take 1 part of the mixed powder and add 8 times the mass of pure water, soak for 30 minutes, heat and reflux for 1 hour, filter, add 6 times the mass of pure water to the residue, heat and reflux for 0.5 hours, filter, combine the two filtrates, concentrate under reduced pressure, dry, and pulverize to obtain the water extract powder of traditional Chinese medicine.

[0043] S3. Take another portion of the mixed powder, add 8 times the mass of 40wt% ethanol solution, extract ultrasonically for 10 min at 400W and 60℃, filter, add 3 times the mass of 60wt% ethanol solution to the filter residue, heat and reflux for 2 h, filter, combine the two filtrates, recover the ethanol, concentrate under reduced pressure, dry, and pulverize to obtain the Chinese herbal alcohol extract powder.

[0044] S4. Mix the water extract powder and alcohol extract powder of traditional Chinese medicine evenly to obtain the traditional Chinese medicine extract.

[0045] Preparation Example 2: Traditional Chinese Medicine Extracts

[0046] The preparation method is as follows:

[0047] S1. Weigh out peony bark, dictamnus root bark, and sophora flavescens in a mass ratio of 3:5:4, pulverize them, pass them through a 40-mesh sieve to obtain a mixed powder, and divide it into two equal portions.

[0048] S2. Take 1 part of the mixed powder and add 12 times the mass of pure water, soak for 60 minutes, heat and reflux for 2 hours, filter, add 10 times the mass of pure water to the filter residue, heat and reflux for 1 hour, filter, combine the two filtrates, concentrate under reduced pressure, dry, and pulverize to obtain the water extract powder of traditional Chinese medicine.

[0049] S3. Take another portion of the mixed powder, add 10 times the mass of 60wt% ethanol solution, ultrasonically extract for 30 min at 200W and 40℃, filter, add 5 times the mass of 80wt% ethanol solution to the filter residue, heat and reflux for 1 h, filter, combine the two filtrates, recover the ethanol, concentrate under reduced pressure, dry, and pulverize to obtain the Chinese herbal alcohol extract powder.

[0050] S4. Mix the water extract powder and alcohol extract powder of traditional Chinese medicine evenly to obtain the traditional Chinese medicine extract.

[0051] Preparation Example 3: Traditional Chinese Medicine Extracts

[0052] The preparation method is as follows:

[0053] S1. Weigh out peony bark, dictamnus root bark, and sophora flavescens in a mass ratio of 2:4:3, pulverize them, pass them through a 40-mesh sieve to obtain a mixed powder, and divide it into two equal portions.

[0054] S2. Take 1 part of the mixed powder and add 10 times the mass of pure water, soak for 45 minutes, heat and reflux for 1.5 hours, filter, add 8 times the mass of pure water to the residue, heat and reflux for 1 hour, filter, combine the two filtrates, concentrate under reduced pressure, dry, and pulverize to obtain the water extract powder of traditional Chinese medicine.

[0055] S3. Take another portion of the mixed powder, add 9 times the mass of 50wt% ethanol solution, extract ultrasonically for 20 min at 300W and 50℃, filter, add 4 times the mass of 70wt% ethanol solution to the filter residue, heat and reflux for 1.5 h, filter, combine the two filtrates, recover the ethanol, concentrate under reduced pressure, dry, and pulverize to obtain the Chinese herbal alcohol extract powder.

[0056] S4. Mix the water extract powder and alcohol extract powder of traditional Chinese medicine evenly to obtain the traditional Chinese medicine extract.

[0057] Comparative Preparation Example 1

[0058] The only difference from Preparation Example 3 is that peony bark was not added to the raw materials. The raw materials were dictamnus root bark and sophora flavescens root bark in a mass ratio of 4:3.

[0059] Comparative Preparation Example 2

[0060] The only difference from Preparation Example 3 is that Dictamnus dasycarpus root bark was not added to the raw materials. The raw materials were Paeonia suffruticosa root bark and Sophora flavescens root bark in a mass ratio of 2:3.

[0061] Comparative preparation example 3

[0062] The only difference from Preparation Example 3 is that Sophora flavescens was not added to the raw materials. The raw materials were Paeonia suffruticosa root bark and Dictamnus dasycarpus root bark in a mass ratio of 2:4.

[0063] Preparation Example 4: Antifungal Compound

[0064] Synthesis route:

[0065]

[0066] The preparation method is as follows:

[0067] T1. 0.01 mol of 2,4-bis(4-morpholino)-6-chloropyrimidine was added to 100 mL of isopropanol, followed by 0.011 mol of m-aminophenylacetylene. The mixture was heated under reflux and stirred for 3 h, then cooled to 0 °C and stirred for 30 min. The mixture was filtered, washed, and dried to obtain intermediate 1 with a yield of 72.4%.

[0068] ESI-MS calculated value: C 20 H 24 N5O2(M+H) + 366.43, measured value: 366.4.

[0069] 1 H NMR (300MHz, CDCl3) δ6.92(m,1H),6.74(d,J=6.3Hz,1H),6.65(s,1H),6.47(d,J=6.0 Hz,1H),4.71(s,1H),4.02(br,1H),3.6-3.62(m,8H),3.02(s,1H),2.9-2.92(m,8H).

[0070] T2. Mix 0.01 mol intermediate 1, 0.013 mol 2,4-dibromobenzyl azide, 10 mL tert-butanol, 10 mL water, 15 mL tetrahydrofuran, 0.25 g copper sulfate, and 0.5 g ascorbic acid thoroughly. Heat to 70 °C and stir for 4 h. Add 25 mL dichloromethane, filter, extract the aqueous phase with dichloromethane, combine the organic phases, dry, concentrate, and purify to obtain the product with a yield of 62.5%.

[0071] ESI-MS calculated value: C 21 H 26 N7O2(M+H) +408.47, measured value: 408.5.

[0072] 1 H NMR (300MHz, CDCl3) δ7.6(s,1H),7.05-7.11(m,5H),5.89(s,1H),4.92(s,2H),3.66-3.70(m,8H),2.9-2.93(m,8H).

[0073] Test Example 1

[0074] The antifungal compound, fluconazole, and ketoconazole obtained in Preparation Example 4 were dissolved in dimethyl sulfoxide to prepare solutions. Before preparing the drug sensitivity plate, the solutions were added to RPMI 1640 culture medium and thoroughly shaken to mix.

[0075] Microsporum canis (ATCC36299), Microsporum gypseum (ATCC14683), Epidermophyton floccosum (ATCC52066), and Malassezia (ATCC44344) were selected as the bacteria to be tested.

[0076] The minimum inhibitory concentrations (MICs) of the two drugs were tested, and the results are shown in Table 1.

[0077] Table 1

[0078]

[0079] As shown in the table above, the antifungal compound prepared in Example 4 of this invention has a good inhibitory effect on some dermatophyte fungi, and its effect is better than that of fluconazole and ketoconazole.

[0080] Preparation Example 5: Active Component

[0081] The herbal extract obtained in Preparation Example 1 and the antifungal compound obtained in Preparation Example 4 were mixed evenly at a mass ratio of 50:1 to obtain the active component.

[0082] Preparation Example 6

[0083] The herbal extract obtained in Preparation Example 1 and the antifungal compound obtained in Preparation Example 4 were mixed evenly at a mass ratio of 100:1 to obtain the active component.

[0084] Preparation Example 7

[0085] The herbal extract obtained in Preparation Example 1 and the antifungal compound obtained in Preparation Example 4 were mixed evenly at a mass ratio of 70:1 to obtain the active component.

[0086] Comparative preparation example 4

[0087] The only difference from Preparation Example 7 is that the antifungal compound was replaced by an equal mass of fluconazole.

[0088] Comparative preparation example 5

[0089] The only difference from Preparation Example 7 is that the antifungal compound was replaced by an equal mass of ketoconazole.

[0090] Test Example 2

[0091] The active components obtained from Preparation Examples 5-7 and Comparative Preparation Examples 4-5 were prepared into solutions. Drug-resistant Candida albicans (ATCC10231) was selected as the test strain.

[0092] The preparation method of drug-resistant Candida albicans is as follows:

[0093] Selected and purified Candida albicans were inoculated into 10 mL of fluconazole-free YEPD liquid medium and cultured with shaking at 30°C. The culture was continued until the bacterial count reached 10⁻⁶. 8 When the cfu / mL level is reached, take 0.1 mL of bacterial culture and add it to 10 mL of YEPD liquid medium containing 16 mg / L of drug (fluconazole, ketoconazole, or an antifungal compound). Continue adding the culture until the bacterial count reaches 10⁻⁶. 8 When the concentration of cfu / mL was reached, another 0.1 mL of bacterial culture was transferred to YEPD medium containing the same concentration of drug and cultured for a further period. Simultaneously, each generation of induced strains was stored at -80°C with 30% glycerol (v / v) and their MICs were determined. By the 8th generation, the MIC of fluconazole reached 140 mg / L, the MIC of ketoconazole reached 125 mg / L, and the MIC of the antifungal compound reached 120 mg / L.

[0094] The MIC results of traditional Chinese medicine extracts and drugs (fluconazole, ketoconazole, or antifungal compounds) against drug-resistant Candida albicans were tested (Preparation Examples 5-7 and Comparative Preparation Examples 4-5). The results are shown in Table 2.

[0095] Table 2

[0096]

[0097] As shown in the table above, among the active components prepared in Examples 5-7 of the present invention, the traditional Chinese medicine extract has a good resistance reversal effect against antifungal compounds. At the same time, it also has a certain resistance reversal effect against fluconazole and ketoconazole, but the effect is not as good as that against antifungal compounds. Therefore, the active components prepared in Examples 5-7 of the present invention have a good antifungal resistance effect.

[0098] Example 1: Antifungal dermatitis agent (spray) containing Danshen, Baishen, and Kushen.

[0099] Composition (mass percentage): 5 wt% of the herbal extract obtained in Preparation Example 1, 0.1 wt% of the antifungal compound obtained in Preparation Example 4, 20 wt% of ethanol, 5 wt% of propylene glycol, 1 wt% of polysorbate-80, 0.1 wt% of sodium benzoate, 2 wt% of glycerol, and the balance being purified water.

[0100] Preparation method: Add the Chinese herbal extract and antifungal compound to ethanol, stir well, add propylene glycol, sodium benzoate, polysorbate-80 and glycerin, stir well, add purified water, stir well, filter with a 0.45μm microporous membrane, and fill into bottles to obtain the product.

[0101] Example 2: Antifungal dermatitis agent (spray) containing Danshen, Baishen, and Kushen.

[0102] Composition (mass percentage): 10 wt% of the herbal extract obtained in Preparation Example 2, 0.1 wt% of the antifungal compound obtained in Preparation Example 4, 40 wt% of ethanol, 10 wt% of propylene glycol, 3 wt% of polysorbate-80, 0.2 wt% of sodium benzoate, 5 wt% of glycerol, and the balance of purified water.

[0103] The preparation method is the same as in Example 1.

[0104] Example 3: Antifungal dermatitis agent (spray) containing Danshen, Baishen, and Kushen.

[0105] Composition (mass percentage): 7 wt% of the herbal extract obtained in Preparation Example 3, 0.1 wt% of the antifungal compound obtained in Preparation Example 4, 30 wt% of ethanol, 7 wt% of propylene glycol, 2 wt% of polysorbate-80, 0.15 wt% of sodium benzoate, 3 wt% of glycerol, and the balance of purified water.

[0106] Comparative Example 1

[0107] The only difference from Example 3 is that the herbal extract was prepared by Comparative Preparation Example 1.

[0108] Comparative Example 2

[0109] The only difference from Example 3 is that the herbal extract was prepared from Comparative Preparation Example 2.

[0110] Comparative Example 3

[0111] The only difference from Example 3 is that the herbal extract was prepared using the comparative preparation example 3.

[0112] Comparative Example 4

[0113] The only difference from Example 3 is that the antifungal compound was replaced by an equal mass of fluconazole.

[0114] Comparative Example 5

[0115] The only difference from Example 3 is that the antifungal compound was replaced by an equal mass of ketoconazole.

[0116] Test Example 3

[0117] Twenty-four 6-week-old Kunming mice (19-20g each) were randomly divided into six groups: mice with damaged skin (Groups 1, 2, and 3 of Example 1) and mice with intact skin (Groups 1, 2, and 3 of Example 1), with four mice in each group, and their sexes were randomly assigned. Forty-eight hours before drug administration, hair was removed from both sides of the spine as a reference line, maintaining symmetry as much as possible, for a bilateral self-control experiment. In the group with damaged skin, the shaved area was slightly incised with a sterile needle, with slight bleeding as the standard for incision. 2 mL of the antifungal dermatitis agent containing Danshen and Kushen prepared in Examples 1-3 was applied to the left side, and physiological saline was evenly applied to the right side. In the group with intact skin, 2 mL of the antifungal dermatitis agent containing Danshen and Kushen prepared in Examples 1-3 was evenly applied to the left side, and physiological saline was evenly applied to the right side. After application, the incision was covered with sterile gauze and secured with adhesive tape. At 1, 12, 24, 48, and 72 hours after administration, the presence of erythema, edema, or other inflammatory symptoms at the application site was observed visually and recorded according to the skin irritation reaction scoring criteria. The results are shown in Table 3.

[0118] Table 3

[0119]

[0120] Note: Skin irritation is determined based on the presence or absence of abnormal skin symptoms. "-" indicates completely normal skin, "+" indicates slightly abnormal skin, and "++" indicates a strong allergic reaction to irritation.

[0121] Test Example 4

[0122] New Zealand white rabbits were randomly selected and divided into a control group, an infection test group, Examples 1-3, and Comparative Examples 1-5, with 10 rabbits in each group. The white fur on the backs of the New Zealand white rabbits was trimmed to approximately 0.5 cm in length using clippers. An area of ​​approximately 2 cm × 2 cm was measured and marked. This area was disinfected with 75% alcohol, and after drying, 1 mL of a 10% alcohol solution was added. 8 A CFU / mL solution of *Microsporum canis* (ATCC11621) was applied evenly with a sterile brush. Control group New Zealand white rabbits were treated with the same dose of sterile PBS. Each experimental group of New Zealand white rabbits had 2 mL of a prepared antifungal dermatology agent containing *Sophora flavescens* sprayed onto the bald area once daily. The treatment was continued for 7 days, and skin lesions were scored after 50 days (scoring criteria are shown in Table 4). Results are shown in Table 5.

[0123] Table 4

[0124]

[0125] Table 5

[0126]

[0127] As shown in the table above, the antifungal dermatological agents containing Danshen and Sophora flavescens prepared in Examples 1-3 of this invention have a good effect on improving the symptoms of skin diseases caused by the fungus Microsporum canis.

[0128] Scanning electron microscopy was performed on severely infected areas of one New Zealand white rabbit from the control group, the infection test group, and the Example 3 group (the control group was randomly selected). The results are shown in the figure. Figure 1 A is the control group (healthy group). No hyphae or spore structures were observed on the skin epidermis. The epidermal cell surface was smooth and intact, appearing as multiple layers of overlapping sheets. Keratin was visible on the fur surface. B is the infection test group (lesion group). Hyphae were present in the skin epidermis, but hyphae damage was relatively obvious. Most cell walls showed obvious collapse and wrinkling. No typical spore structures were observed. Epidermal cells were wrinkled and partially damaged. C is the Example 3 group (drug administration group). No hyphae or spore structures were observed on the skin epidermis. The epidermal cell surface was smooth and intact, appearing as multiple layers of overlapping sheets. A very few cells were damaged.

[0129] Example 4: Antifungal dermatitis agent (ointment) containing Danshen, Baishen, and Kushen.

[0130] Composition (mass percentage): 7.5-15 wt% of the herbal extract obtained in Preparation Example 3, 0.15 wt% of the antifungal compound obtained in Preparation Example 4, 15-25 wt% of white petrolatum, 5-10 wt% of glyceryl monostearate, 10-15 wt% of liquid paraffin, 3-5 wt% of lanolin, 0.1-0.2 wt% of ethylparaben, and the balance being purified water.

[0131] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An antifungal dermatological agent containing Danshen, Baishen, and Kushen, characterized in that, The product includes an active ingredient and other pharmaceutically acceptable excipients. The active ingredient comprises a traditional Chinese medicine extract and an antifungal compound in a mass ratio of (50-100):

1. The traditional Chinese medicine extract is obtained by extracting peony bark, dictamnus root bark, and sophora flavescens root in a mass ratio of (1-3):(2-5):(2-4). The antifungal compound has the structural formula shown in Formula I or a pharmaceutically acceptable salt thereof. Formula I.

2. The antifungal dermatological agent containing Danshen and Sophora flavescens according to claim 1, characterized in that, The preparation method of the traditional Chinese medicine extract is as follows: S1. Weigh out the peony bark, dictamnus root bark, and sophora flavescens according to the proportion, crush them, pass them through a 40-mesh sieve to obtain a mixed powder, and divide it into 2 equal portions. S2. Take 1 part of the mixed powder and add 8-12 times the mass of pure water, soak for 30-60 minutes, heat and reflux to extract for 1-2 hours, filter, add 6-10 times the mass of pure water to the filter residue, heat and reflux to extract for 0.5-1 hours, filter, combine the two filtrates, concentrate under reduced pressure, dry, and pulverize to obtain the water extract powder of traditional Chinese medicine. S3. Take another portion of the mixed powder, add 8-10 times the mass of 40-60wt% ethanol solution, extract by ultrasonication for 10-30 min, filter, add 3-5 times the mass of 60-80wt% ethanol solution to the filter residue, heat and reflux for 1-2 h, filter, combine the two filtrates, recover the ethanol, concentrate under reduced pressure, dry, and pulverize to obtain the Chinese herbal alcohol extract powder. S4. Mix the water extract powder and alcohol extract powder of traditional Chinese medicine evenly to obtain the traditional Chinese medicine extract.

3. The antifungal dermatological agent containing Danshen and Sophora flavescens according to claim 2, characterized in that, The ultrasonic extraction power is 200-400W, and the temperature is 40-60℃.

4. The antifungal dermatological agent containing Danshen and Sophora flavescens according to claim 1, characterized in that, The method for preparing the antifungal compound is as follows: T1. 2,4-bis(4-morpholino)-6-chloropyrimidine was reacted with m-aminophenylacetylene to prepare intermediate 1, with the following structure: ; T2. Intermediate 1 was reacted with 2,4-dibromobenzyl azide to obtain the product.

5. The antifungal dermatological agent containing Danshen and Sophora flavescens according to claim 4, characterized in that, The molar ratio of 2,4-bis(4-morpholino)-6-chloropyrimidine to m-aminophenylacetylene in step T1 is 1:1-1.

2.

6. The antifungal dermatitis agent containing Danshen and Sophora flavescens according to claim 4, characterized in that, The molar ratio of intermediate 1 and 2,4-dibromobenzyl azide in step T2 is 1:1.2-1.

5.

7. The antifungal dermatological agent containing Danshen and Sophora flavescens according to claim 1, characterized in that, The dosage form of the medicine is at least one of spray and ointment.

8. A method for preparing an antifungal dermatological agent containing *Salvia miltiorrhiza* and *Sophora flavescens* as described in any one of claims 1-7, characterized in that, After mixing the extract of traditional Chinese medicine and the antifungal compound evenly, the active component is obtained. Excipients are added, and the mixture is prepared according to conventional formulation methods to obtain an antifungal dermatology agent containing Danshen, Baishen, and Kushen.

9. The use of an antifungal dermatological agent containing Danshen and Sophora flavescens as described in any one of claims 1-7 in the preparation of veterinary drugs.

10. The use of an antifungal dermatitis agent containing Danshen and Sophora flavescens as described in any one of claims 1-7 in the preparation of a veterinary drug with antifungal effects on cats and dogs.

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