Traditional Chinese medicine composition and use thereof in preparing medicine for inhibiting fungi

Abstract

This invention relates to the field of traditional Chinese medicine technology, and more particularly to a traditional Chinese medicine composition and its application in the preparation of antifungal drugs. The composition provided by this invention comprises Dioscorea hypoglauca, Cnidium monnieri, Bistorta officinalis, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum, and Terminalia chebula. This composition has the effect of inhibiting the growth of superficial dermatophytes (such as Trichophyton rubrum and Candida albicans), which helps to reduce the occurrence of superficial dermatophyte infections and has the potential to be developed into a drug for treating superficial dermatophyte infections.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to traditional Chinese medicine compositions and their application in the preparation of drugs for inhibiting fungi. Background Technology

[0002] Fungal skin diseases are infectious diseases of the skin, mucous membranes, hair, and nails caused by fungi. Also known as superficial fungal infections, they are the most common infectious diseases in dermatology. Due to their high incidence and recurrence rates, poor patient compliance, the toxic side effects of Western medicines, and the emergence of drug-resistant fungal strains, patients' quality of life is severely affected.

[0003] Based on the location of the dermatophyte infection, fungal skin diseases can be classified into tinea capitis (dermatophyte infection of the scalp), tinea corporis (dermatophyte infection of the smooth skin of the body), tinea cruris (dermatophyte infection of the groin), tinea manuum (dermatophyte infection of the hands), tinea pedis (dermatophyte infection of the feet, also known as athlete's foot), and onychomycosis (dermatophyte infection of the nails), etc. Superficial fungal infections are mainly caused by dermatophytes, with other common pathogens including Candida and Malassezia. The main pathogens of dermatophytes originate from three genera: Trichophyton, Epidermophyton, and Microsporum, with Trichophyton rubrum accounting for more than 60% of dermatophyte infections. Candida albicans, a type of yeast, is also one of the most common causes of skin fungal infections. Warm, moist, or sweaty areas provide a favorable environment for yeast growth, including the armpits, groin, skin between the fingers and toes, corners of the mouth, and the area under the breasts.

[0004] Clinically, allylamine and azole antifungal drugs are commonly used. However, the use of these antibiotics can lead to a range of adverse reactions, including liver and kidney damage, allergic reactions, gastrointestinal issues, and bone marrow toxicity. Accurate diagnosis, identification of the causative pathogen and the stage of disease development, selection of appropriate drug type, dosage form, dosage, and course of treatment, and avoiding indiscriminate prophylactic medication while employing rational combination therapy are crucial for improving treatment efficiency. This makes the discovery and identification of new pathogens and clues from synthetic chemicals or natural products an urgent task.

[0005] Compared to synthetic chemicals, traditional Chinese medicine (TCM) has advantages such as structural diversity and lower toxicity. TCM provides a potential source for antifungal drugs, whether in its nascent form or as original templates for structural optimization, to obtain more effective and safer derivatives, which is of great significance for the development of treatments for superficial fungal infections of the skin. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a traditional Chinese medicine composition and its application in the preparation of a drug for inhibiting fungi.

[0007] The traditional Chinese medicine composition provided by this invention is prepared from Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine, and Terminalia chebula.

[0008] In some embodiments, the mass ratio of the following herbs is (10-20):(8-15):(8-15):(10-15):(9-12):(10-15):(12-20):(9-12).

[0009] In some specific embodiments, the mass ratio of the following herbs is 10:8:15:11:12:12:15:12;

[0010] In other specific embodiments, the mass ratio of the following herbs is 10:15:12:10:9:10:20:9.

[0011] In other specific embodiments, the mass ratio of the following herbs is 15:10:10:12:9:15:15:9;

[0012] In other specific embodiments, the mass ratio of Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine, and Terminalia chebula is 15:9:12:12:12:12:12:11;

[0013] In other specific embodiments, the mass ratio of the following herbs is 12:9:15:15:9:12:14:9.

[0014] In other specific embodiments, the mass ratio of the following herbs is 20:8:8:12:12:10:15:10.

[0015] The traditional Chinese medicine composition provided by this invention has the effect of inhibiting common superficial fungal infections of the skin, especially inhibiting the growth of Trichophyton rubrum and Candida albicans, which helps to reduce the formation of superficial fungal infections of the skin and has the potential to be developed into a drug for treating superficial fungal infections of the skin.

[0016] The traditional Chinese medicine composition of this invention is prepared by mixing the following herbs: Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine, and Terminalia chebula. Alternatively, it can be prepared by further processing, preparation, and / or extraction of the aforementioned herbs; this invention does not limit the specific methods used. During the extraction process, each herb can be extracted individually, or two or more herbs can be extracted in combination; this invention also does not limit the methods used.

[0017] In some embodiments, the present invention also provides a method for preparing the traditional Chinese medicine composition as described above, comprising:

[0018] The Chinese herbal composition is obtained by extracting the following herbs with 70%–95% ethanol: Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine, and Terminalia chebula. The extracts are then concentrated, dried, and mixed to obtain the Chinese herbal composition.

[0019] In some embodiments, the ethanol is an aqueous solution of ethanol, wherein the volume fraction of ethanol is 70% to 95%, preferably 80% to 90%. For example, the volume fraction of ethanol is 80%, 85%, or 90%.

[0020] In some embodiments, the volume ratio of ethanol to medicinal materials is (10-20):1. Preferably, the volume ratio of ethanol to medicinal materials is 10:1, 15:1 or 20:1.

[0021] In some embodiments, the extraction method is standard Soxhlet extraction.

[0022] Furthermore, this invention also provides the application of at least one of the following: *Dioscorea hypoglauca*, *Cnidium monnieri*, *Polygonum bistorta*, *Dioscorea nipponica*, *Murraya paniculata*, *Dioscorea opposita*, *Polygonum multiflorum*, and *Terminalia chebula* in the preparation of antifungal drugs. Experiments show that extracts obtained from any one of these ingredients, after extraction, possess antifungal properties. Mixtures obtained by mixing these ingredients in pairs, or by mixing three or more of them, also possess antifungal properties.

[0023] Furthermore, the present invention also provides the use of the traditional Chinese medicine composition as described above, or the traditional Chinese medicine composition prepared by the preparation method as described above, in the preparation of a drug for inhibiting fungi.

[0024] In this invention, the fungus is a pathogen causing superficial fungal infections.

[0025] The traditional Chinese medicine composition provided by this invention can inhibit the pathogens causing various superficial fungal infections. In the embodiments of this invention, *Trichophyton rubrum* and / or *Candida albicans* are used as examples to verify the antibacterial effect of the traditional Chinese medicine or the traditional Chinese medicine composition.

[0026] The present invention also provides a drug for preventing and treating superficial fungal infections, comprising at least one of the following: Dioscorea hypoglauca, Cnidium monnieri, Bistorta officinalis, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum, and Terminalia chebula, or a traditional Chinese medicine composition as described above, or a traditional Chinese medicine composition prepared by the method described above.

[0027] The medicament described in this invention also includes excipients. The excipients are selected according to the dosage form and formulation of the medicament. For example, the dosage form is a topical dosage form, such as an ointment, emulsion, lotion, powder, cream, liniment, or gel. For example, the excipients are selected from: purified water, oils, lipids, higher alcohols, or waxes, etc. Alternatively, they are selected from: surfactants, humectants, antioxidants, preservatives, thickeners, chelating agents, pigments, pH adjusters, fragrances, etc.

[0028] In some embodiments, the dosage form of the drug is an ointment, cream, or gel, and the excipients are selected from: alginic acid, gelatin, hyaluronic acid, animal oil, vegetable oil, wax, paraffin, starch, cellulose derivatives, polyethylene glycol, organosilicon, bentonite, silica, talc, zinc oxide, etc.

[0029] In some embodiments, the dosage form of the drug is a powder, and the excipients are selected from lactose, talc, silicon dioxide, aluminum hydroxide, calcium silicate, or polyamide.

[0030] In some embodiments, the dosage form of the drug is a solution, lotion, or emulsion, and the excipients are selected from solvents, solubilizers, or emulsifiers. Examples include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylethylene glycol oil, glycerol aliphatic esters, polyethylene glycol, or fatty acid esters of sorbitol.

[0031] The drug described in this invention can be used directly or after dilution. This invention does not limit the use of the drug. Preferably, the content of traditional Chinese medicine or traditional Chinese medicine composition in the drug described in this invention is 10 wt%.

[0032] Furthermore, the present invention also provides a method for preventing and treating superficial fungal infections, which includes administering the drug described in the present invention, wherein the administration of the drug described in the present invention includes spraying, applying, or plastering.

[0033] This invention provides a traditional Chinese medicine composition comprising Dioscorea hypoglauca, Cnidium monnieri, Bistorta officinalis, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine, and Terminalia chebula. This composition has the effect of inhibiting the growth of superficial dermal fungi (such as Trichophyton rubrum and Candida albicans), which helps to reduce the occurrence of superficial dermal fungal infections and has the potential to be developed into a drug for treating superficial dermal fungal infections. Detailed Implementation

[0034] This invention provides a traditional Chinese medicine composition and its application in the preparation of antifungal drugs. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0035] The numerical ranges and parameters involved in this invention have been presented as precisely as possible in the specific embodiments. However, any numerical value inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise explicitly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have a reasonable deviation within a certain range, such as ±10%, ±5%, ±1%, or ±0.5%.

[0036] The embodiments of this invention describe some examples. These examples illustrate certain implementations of the invention. However, this does not mean that the effects of the invention can only be achieved in these examples. In fact, any concentration of each component between the two endpoints shown in Examples 1-6 can achieve good antifungal effects, with Examples 3 and 6 showing the best results, exhibiting significant advantages in antibacterial effect compared to various validated formulations. The comparative examples only list some cases where the effects were not good in the experiments. In addition, many other attempts were made during the research and development process, such as using different components, different ratios, or different extraction processes, but the effects of these attempts were not as good as those of Examples 1-6, and will not be elaborated upon here.

[0037] The test materials used in this invention are all common commercial products and can be purchased on the market.

[0038] Dioscorea septemloba Thunb. or Dioscorea futschauensis Uline ex R.kunth are the dried rhizomes of plants in the Dioscoreaceae family. [Properties and Flavors] Bitter, neutral. [Meridians Entered] Kidney and Stomach meridians. [Functions and Indications] Clears dampness and turbidity, dispels wind and relieves numbness. Used for gonorrhea with leukorrhea, excessive vaginal discharge, damp-heat sores, and lumbago. This invention has found that Dioscorea septemloba extract has an inhibitory effect on common superficial fungal infections of the skin.

[0039] Cnidium monnieri L., a plant belonging to the Apiaceae family, is the fruit of the herb. [Taste and Properties] Pungent; bitter; warm in nature. [Meridians Entered] Spleen; Kidney. [Functions and Indications] Warms the kidneys and strengthens yang; dries dampness and kills parasites; dispels wind and relieves itching. It is used for male impotence; scrotal eczema and itching; female infertility due to cold uterus; leukorrhea due to cold and dampness; vulvar itching and swelling; rheumatic pain; damp sores and scabies. This invention has discovered that Cnidium monnieri extract has an inhibitory effect on common superficial fungal infections of the skin.

[0040] Bistorta bistorta L., a plant in the Polygonaceae family, is a dried rhizome. [Properties and Flavors] Bitter, astringent, slightly cold. [Meridians Entered] Lung, Liver, Large Intestine. [Functions and Indications] Clears heat and detoxifies, reduces swelling, and stops bleeding. Used for dysentery, diarrhea due to heat, cough due to lung heat, carbuncles, scrofula, oral ulcers, hematemesis, epistaxis, hemorrhoidal bleeding, and snake bites. This invention has discovered that Bistorta bistorta extract has an inhibitory effect on common superficial fungal infections of the skin.

[0041] Dioscorea nipponica Makino, a plant belonging to the genus Dioscorea in the family Dioscoreaceae, is used medicinally for its rhizome. [Properties and Flavors] Bitter, neutral. [Functions and Indications] Relieves muscle tension, dispels wind and alleviates pain. Used for rheumatic pain, rheumatic joint pain, numbness of muscles and bones, Kashin-Beck disease, traumatic injuries, and bronchitis. This invention has discovered that Dioscorea nipponica extract has an inhibitory effect on common superficial fungal infections of the skin.

[0042] Murraya exotica L. and Murraya paniculata (L.) Jack, both belonging to the Rutaceae family, are the dried leaves and leafy young branches of these plants. [Taste and Properties] Pungent; slightly bitter; warm in nature; slightly toxic. [Meridians Entered] Heart; Liver; Lung. [Functions and Indications] Promotes qi and blood circulation; disperses blood stasis and relieves pain; detoxifies and reduces swelling. Mainly used for stomach pain; traumatic swelling and pain; sores and carbuncles; snake and insect bites. This invention has discovered that Murraya exotica extract has an inhibitory effect on common superficial fungal infections of the skin.

[0043] Huangshan yam is the rhizome of *Dioscorea panthaica* Prain et Burk., a plant belonging to the genus *Dioscorea* of the family Dioscoreaceae. It is harvested in autumn, washed, and sun-dried. [Properties and Flavors] Sweet, slightly pungent, neutral. [Functions and Indications] Detoxifies, reduces swelling, and relieves pain. Used for stomach pain and traumatic injuries; externally used to treat lymph node tuberculosis. This invention has discovered that Huangshan yam extract has an inhibitory effect on common superficial fungal infections of the skin.

[0044] Polygonum multiflorum Thunb., a plant of the Polygonaceae family, is a dried vine stem. [Properties and Flavors] Sweet, neutral. [Meridians Entered] Heart and Liver meridians. [Functions and Indications] Nourishes blood and calms the mind, dispels wind and unblocks meridians. Used for insomnia, excessive dreaming, blood deficiency and body pain, rheumatic pain; externally for pruritus. This invention has discovered that extracts from Polygonum multiflorum vine have an inhibitory effect on common superficial fungal infections of the skin.

[0045] Terminalia chebula is the dried, ripe fruit of *Terminalia billerica* (Gaertn.) Roxb., a plant in the Combretaceae family. [Properties and Flavors] Sweet, astringent, neutral. [Functions and Indications] Clears heat and detoxifies, astringes and nourishes blood, harmonizes other herbs. Used for various febrile diseases, diarrhea, jaundice, liver and gallbladder diseases, and post-illness weakness. This invention has discovered that Terminalia chebula extract has an inhibitory effect on common superficial fungal infections of the skin.

[0046] The traditional Chinese medicine composition provided by this invention comprises *Smilax glabra*, *Cnidium monnieri*, *Polygonum bistorta*, *Dioscorea nipponica*, *Murraya paniculata*, *Dioscorea opposita*, *Polygonum multiflorum* vine, and *Terminalia chebula*. Alternatively, the traditional Chinese medicine composition provided by this invention comprises extracts of *Smilax glabra*, *Cnidium monnieri*, *Polygonum bistorta*, *Dioscorea nipponica*, *Murraya paniculata*, *Dioscorea opposita*, *Polygonum multiflorum* vine, and *Terminalia chebula*. The traditional Chinese medicine extract composition of this invention can prevent and treat superficial fungal infections of the skin, especially infections caused by fungi commonly found in superficial skin diseases, such as *Trichophyton rubrum* and *Candida albicans*.

[0047] In this embodiment of the invention, MIC refers to the minimum inhibitory concentration, which is the minimum concentration of antibacterial drug required to completely inhibit bacterial growth in the culture flask.

[0048] The present invention will be further illustrated below with reference to the embodiments:

[0049] Preparation of medicinal extracts and compositions in the examples

[0050] 1. Preparation of Dioscorea hypoglauca extract: Dioscorea hypoglauca was pulverized into powder. 10g of the powder was placed on filter paper and wrapped into a packet. The packet was then placed at the bottom of a cylindrical glass sample tube in a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the final Dioscorea hypoglauca extract.

[0051] 2. Preparation of Cnidium monnieri extract: Cnidium monnieri was pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. This packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the final Cnidium monnieri extract.

[0052] 3. Preparation of Polygonum bistorta extract: Polygonum bistorta medicinal material was pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. The packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the final Polygonum bistorta extract.

[0053] 4. Preparation of *Dioscorea nipponica* extract: *Dioscorea nipponica* herb was pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. This packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the final *Dioscorea nipponica* extract.

[0054] 5. Preparation of Murraya paniculata extract: Murraya paniculata herb was pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. The packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the final Murraya paniculata extract.

[0055] 6. Preparation of Huangshan (Dioscorea opposita) extract: Huangshan medicinal materials were pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. This packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was subjected to vacuum low-temperature freeze-drying to obtain the final Huangshan extract.

[0056] 7. Preparation of Polygonum multiflorum vine extract: Polygonum multiflorum vine was pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. The packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the final Polygonum multiflorum vine extract.

[0057] 8. Preparation of Terminalia chebula extract: Terminalia chebula medicinal material was pulverized into powder. 10g of the powder was weighed, placed on filter paper, and wrapped into a packet. The packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor for standard Soxhlet extraction. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the Terminalia chebula extract.

[0058] 9. Take extracts of Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum, and Terminalia chebula. Mix the eight kinds of Chinese herbal extracts according to the proportions shown in Table 1 to form Examples 1 to 6, with a total of 95 parts. Then dilute them with RPMI 1640 medium solution as shown in Table 1.

[0059] Table 1:

[0060]

[0061] 10. Preparation of positive drug extract: The positive drug, *Sophora flavescens*, was pulverized to form *Sophora flavescens* powder. 10g of the powder was placed on filter paper and wrapped into a packet. This packet was then placed at the bottom of the cylindrical glass sample tube of a Soxhlet extractor. 15 times the volume of 90% ethanol was poured into a solvent cup and fixed to the lower interface of the Soxhlet extractor. The extraction stage heating level was set to 16, and the extraction time was 3 hours. The drying stage heating level was set to 10, and the drying time was 20 minutes. Soxhlet extraction was then performed. After extraction, the extract was freeze-dried under vacuum at low temperature to obtain the *Sophora flavescens* extract. The *Sophora flavescens* extract was diluted with RPMI 1640 medium solution, and the results were compared with those in the examples.

[0062] 11. The preparation of single Chinese herbal medicines is missing. The proportions shown in Table 2 are as follows.

[0063] Table 2

[0064]

[0065] 12. Experimental Protocol: Extracts of Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum, and Terminalia chebula were taken and tested according to the "Liquid-based Dilution Method for Antifungal Susceptibility Testing of Sporulating Filamentous Fungi" (M38-A) recommended by the National Committee for Clinical Laboratory Standards (NCCLS). The minimum inhibitory concentration (MIC) of the single herbs, different embodiments, and formulas lacking a single herb against fungi was determined to demonstrate the antifungal effect of the formula.

[0066] II. Determination of MIC of extracts from single Chinese medicinal herbs, the specific procedure is as follows:

[0067] 1. Preparation of PDA: Weigh 3.7g of PDA powder, add 100mL of distilled water, dissolve thoroughly, autoclave, and finally dispense into 15×150 test tubes (sterile) for later use;

[0068] 2. Activation of bacterial strains: After incubation on PDA plates at 28°C for 7–14 days, drug sensitivity tests are performed.

[0069] 3. After irradiating a sterile 96-well flat-bottom microplate with UV light for 30 min and cooling, add the drug to wells 1-10 of each row to achieve a working concentration of 100 μL / well. Then add 100 μL of the prepared fungal culture solution. Add 100 μL of liquid culture medium to the negative control well. Well 11 is the negative control (containing only 100 μL of RPMI 1640 medium and 100 μL of drug, without fungal culture solution), and well 12 is the positive control (containing only 100 μL of culture medium and 100 μL of fungal culture solution, without drug). Place the plate on a microplate and shake for 5 min to thoroughly mix all liquids. The final experimental concentrations of the drug, from highest to lowest, are: 500, 250, 125, 62.5, 31.3, 15.6, 7.8, 3.9, 1.9, 1.0 μg / mL.

[0070] 4. Culture conditions and result interpretation: Incubate at 28℃ in a CO2 incubator for 24-48 hours and read the results. In this experiment, whenever the MIC value is read, it should be compared with the following standards and controls without stirring: 0: Clear to the naked eye; 1: Slightly blurred (80% inhibition); 2: Significantly reduced turbidity (50% inhibition); 3: Slightly reduced turbidity; 4: No reduction in turbidity. In this experiment, 1 (slightly blurred) was selected as the endpoint for determination.

[0071] III. Determination of MIC of different formulations in various embodiments, the specific process is as follows:

[0072] 1. The drug extracts from Examples 1 to 6 were mixed and diluted to 1 mg / mL using RPMI 1640 medium.

[0073] 2. The experimental procedures and result interpretation are the same as those in “MIC determination of single herb extracts 1-4”.

[0074] IV. The determination of the antibacterial effect of single Chinese herbal medicines is lacking. The specific process is as follows:

[0075] 1. Mix the drug extracts in ratios 1 to 3 and dilute them to 1 mg / mL using RPMI 1640 medium.

[0076] 2. The experimental procedures and result interpretation are the same as those in “MIC determination of single herb extracts 1-4”.

[0077] V. Determination results of MIC of extracts from single Chinese medicinal herbs

[0078] 1. The determination results of MIC of single herbal extracts are shown in Table 3.

[0079] Table 3

[0080] traditional Chinese medicine fungi MIC (μg / mL) fungi MIC (μg / mL) Smilax china Trichophyton rubrum 31.3 Candida albicans 31.3 Cnidium monnieri Trichophyton rubrum 62.5 Candida albicans 250 Biscuit Trichophyton rubrum 31.3 Candida albicans 500 Pangolin Trichophyton rubrum 15.6 Candida albicans 62.5 Nine-mile fragrance Trichophyton rubrum 125 Candida albicans 250 Yellow yam Trichophyton rubrum 15.6 Candida albicans 15.6 Polygonum multiflorum vine Trichophyton rubrum 125 Candida albicans 125 Terminalia chebula Trichophyton rubrum 125 Candida albicans 500 Sophora flavescens (positive control) Trichophyton rubrum >500 Candida albicans >500 Solvent control (negative control) Trichophyton rubrum >500 Candida albicans >500

[0081] It can be seen that the extracts of eight traditional Chinese medicines showed antibacterial effects against two fungi at concentrations of 500 μg / mL or even lower, and the effects were stronger than those of Sophora flavescens, a commonly used antifungal drug in clinical practice.

[0082] VI. Results of Antibacterial Efficacy Determination of Different Formulations

[0083] 1. The determination results of MIC of different formulations are shown in Table 4.

[0084] Table 4

[0085] Example fungi MIC (μg / mL) fungi MIC (μg / mL) Example 1 Trichophyton rubrum 31.3 Candida albicans 125 Example 2 Trichophyton rubrum 62.5 Candida albicans 250 Example 3 Trichophyton rubrum 15.6 Candida albicans 62.5 Example 4 Trichophyton rubrum 31.3 Candida albicans 125 Example 5 Trichophyton rubrum 15.6 Candida albicans 125 Example 6 Trichophyton rubrum 15.6 Candida albicans 62.5 Solvent control (negative control) Trichophyton rubrum >500 Candida albicans >500

[0086] 2. As can be seen from Tables 3 and 4, all eight Chinese herbal extracts, whether used alone or in combination, have the effect of inhibiting the growth of the two fungi.

[0087] VII. Lack of results for testing the antibacterial effects of single Chinese herbal medicines

[0088] 1. The results of the determination of the antibacterial effect of single Chinese herbal medicines are missing, as shown in Table 5.

[0089] Table 5

[0090] Proportion fungi MIC (μg / mL) fungi MIC (μg / mL) Formula 1 (i.e., Example 6) Trichophyton rubrum 15.6 Candida albicans 62.5 2 ratio Trichophyton rubrum 125 Candida albicans 250 3 ratio Trichophyton rubrum 250 Candida albicans 250 Solvent control (negative control) Trichophyton rubrum >500 Candida albicans >500

[0091] 2. The ratio 1 in Table 5 is the same as Example 6 in Table 4. Among the ratios listed in Table 6, this ratio has the lowest MIC value against both fungi. Based on this, a comparison was set up with the absence of Cnidium monnieri (ratio 2) or Dioscorea hypoglauca (ratio 3). As can be seen from Table 5, the antibacterial effect is significantly reduced when the principal drug (Cnidium monnieri or Dioscorea hypoglauca) of the traditional Chinese medicine formula is missing, indicating that these two drugs are essential for achieving a good antibacterial effect.

[0092] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for inhibiting Trichophyton rubrum and / or Candida albicans, characterized in that, It is prepared from Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine and Terminalia chebula; the mass ratio of Dioscorea hypoglauca, Cnidium monnieri, Polygonum bistorta, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine and Terminalia chebula is (10~20):(8~15):(8~15):(10~15):(9~12):(10~15):(12~20):(9~12).

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The mass ratio of the following herbs is 10:8:15:11:12:12:15:12; Or the mass ratio of the following herbs is 10:15:12:10:9:10:20:9; Or the mass ratio of the following herbs is 15:10:10:12:9:15:15:9; Or the mass ratio of the following herbs is 15:9:12:12:12:12:12:11:11; Or the mass ratio of the following herbs is 12:9:15:15:9:12:14:9; Or the mass ratio of the following herbs is 20:8:8:12:12:10:15:

10.

3. The method for preparing the traditional Chinese medicine composition according to claim 1 or 2, characterized in that, include: The Chinese herbal composition is obtained by extracting the following herbs with 70% to 95% ethanol: Dioscorea hypoglauca, Cnidium monnieri, Bistorta officinalis, Dioscorea nipponica, Murraya paniculata, Dioscorea opposita, Polygonum multiflorum vine, and Terminalia chebula. The extracts are then concentrated, dried, and mixed to obtain the Chinese herbal composition.

4. The preparation method according to claim 3, characterized in that, The extraction was a Soxhlet extraction, with an ethanol-to-medicinal-material volume ratio of 15:

1.

5. The traditional Chinese medicine composition according to claim 1 or 2, or the traditional Chinese medicine composition prepared by the preparation method according to claim 3 or 4, in the preparation of a drug for inhibiting Trichophyton rubrum and / or Candida albicans.

6. A drug for preventing and treating infections caused by Trichophyton rubrum and / or Candida albicans, characterized in that, This includes the traditional Chinese medicine composition according to claim 1 or 2, or the traditional Chinese medicine composition prepared by the preparation method according to claim 3 or 4.

7. The drug according to claim 6, characterized in that, The dosage form of the drug is an ointment, gel, or lotion.

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