New application of traditional Chinese medicine composition

Abstract

The invention relates to the field of traditional Chinese medicine, in particular to application of a traditional Chinese medicine composition in preparation of medicine for preventing and treating diabetes mellitus complicated with candida infection.The traditional Chinese medicine composition is prepared from, by weight, 8-25 parts of oriental wormwood, 8-25 parts of solidago decurrens, 8-25 parts of duckweed, 8-25 parts of herba lycopi and 6-15 parts of horsetail, and candida is candida auricula or candida albicans.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and in particular relates to a new use of a traditional Chinese medicine composition. Background Art

[0002] Diabetes is mainly a metabolic disorder characterized by abnormal blood sugar metabolism. The series of changes in metabolic levels caused by it can lead to oxidative environmental disorders, which in turn lead to multiple complications in patients. Poor blood sugar control in diabetic patients increases the risk of skin, bone, eye, ear, gastrointestinal, urinary and respiratory infections, and significantly increases hospitalization and mortality rates. Diabetic fungal infection has become one of the important medical problems that need to be urgently treated in clinical practice. Currently, the antifungal drugs available in clinical practice include azoles, polyenes and echinocandin drugs, and representative drugs include fluconazole, amphotericin B and caspofungin. Amphotericin B has good antibacterial activity and a wide antibacterial spectrum, but the drug has severe nephrotoxicity, which limits its clinical use. Fluconazole, which is currently the most widely used in clinical practice, only has an antibacterial effect. In long-term treatment, fungal infection recurrence and drug resistance are serious. After long-term treatment, the resistance rate of Candida albicans isolated from clinical practice to fluconazole is as high as 40%, which seriously affects the long-term efficacy of patients. Even for the latest new antifungal drug caspofungin, there are increasing reports of Candida resistance to it, and its limited antibacterial spectrum and high price also limit its clinical application. These make the treatment of fungal infections increasingly challenging. In the past decade, Candida auris has emerged and spread globally. It is a hospital fungus that has become a "serious threat" to medical institutions around the world. Candida auris has strong fungal resistance and chronic infections with poor prognosis in immunocompromised patients. Since Candida auris is usually resistant to two or even three clinical antifungal drugs, diabetes combined with Candida auris infection has become a difficult problem that needs to be solved urgently in the clinic. Therefore, the treatment of diabetes combined with Candida auris infection is a key and difficult problem in current clinical treatment and has great clinical significance. Summary of the invention

[0003] The purpose of the present invention is to provide a new use of a Chinese medicine composition.

[0004] The first aspect of the present invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for preventing and treating diabetes combined with Candida infection. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8 to 25 parts of Artemisia capillaris, 8 to 25 parts of Solidago asiatica, 8 to 25 parts of Duckweed, 8 to 25 parts of Zedoariae rhizome, and 6 to 15 parts of Equisetum arvense. The Candida is Candida auris or Candida albicans.

[0005] In a preferred example, the Chinese medicine composition is made of the following raw materials in parts by weight: 15 parts of Artemisia capillaris, 15 parts of Solidago asiatica, 15 parts of Duckweed, 15 parts of Zedoariae rhizome, and 10 parts of Equisetum arvense.

[0006] The details of various aspects of the present invention will be described in detail in the following sections. The features, objectives and advantages of the present invention will become more apparent through the following description and the claims. DETAILED DESCRIPTION

[0007] The traditional Chinese medicine compound of the invention consists of capillary wormwood herb, lycopi herb, common goldenrop, spirodelae herb and equiseti hiemalis herb.

[0008] Among them, Capillary Wormwood Herb is the seedling of the perennial herb Artemisia capillaris. It is bitter, slightly cold, and enters the spleen, stomach, liver and gallbladder meridians. It has the effects of clearing heat, promoting dampness, and relieving jaundice, and is mostly used in the treatment of jaundice.

[0009] Lycopi Herba, the whole herb of the Labiatae plant Dioscorea oleracea or Dioscorea oleracea. It can relieve wind and relieve exterior symptoms, regulate menstruation and activate blood circulation, and detoxify and reduce swelling. It can treat wind-heat cold, fever at the beginning of febrile disease, irregular menstruation, amenorrhea, metrorrhagia, unexplained swelling, heat-toxic sores, and urticaria and itching.

[0010] Common Goldenrop, the whole herb or the whole herb with roots of the plant Common Goldenrop of the Asteraceae family. It is yellow in color, fragrant, neutral in nature, slightly bitter and flat in taste. Common Goldenrop can dispel wind and heat, and has antibacterial and anti-inflammatory properties. It is used for wind-heat cold and headache, (not suitable for wind-cold) to clear away heat and toxic substances, and to relieve swelling and pain. It can be used for upper respiratory tract infection, tonsillitis, sore throat, bronchitis, pneumonia, hemoptysis caused by tuberculosis, acute and chronic nephritis, malnutrition in children, and external use to treat traumatic injuries, snake bites, sores, swelling and toxins, and mastitis.

[0011] Duckweed (Spirodelae Herb), a perennial floating plant of the Araceae family, is mainly used to treat seasonal fever, wind-heat urticaria, skin itching, sores, erysipelas, dispersing wind-heat, rash, diuresis and swelling, measles that do not break out, edema and oliguria. It is used for external wind-heat symptoms, measles that do not break out smoothly, urticaria, skin itching, and edema and external symptoms. It can be used for urticaria and acute nephritis. It can also promote dampness and reduce jaundice, and is used for acute icteric hepatitis and chronic hepatitis.

[0012] Equiseti Hiemalis Herba, the whole herb of the plant Equisetaceae, is used as medicine. Equiseti is sweet and bitter in taste and neutral in nature. It enters the lung and liver meridians and has the effects of dispersing wind-heat, improving eyesight and removing cataracts, and stopping bleeding. It is used to treat cataracts in the eyes, tears in the wind, intestinal wind bleeding, bloody dysentery, liver loss, malaria, sore throat, and carbuncle.

[0013] Based on the clinical application of the Chinese medicine compound, the inventors respectively constructed a normal mouse model of Candida auris infection and a diabetes combined with Candida auris infection model to study the effect of the Chinese medicine compound on Candida auris infection. The research results of the present invention confirm that the Chinese medicine compound has a significant effect in treating diabetes combined with Candida auris infection and can effectively guide clinical medication.

[0014] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually performed under normal conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight.

[0015] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0016] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0017] Example 1 Preparation of Chinese medicine composition

[0018] Take 15g of Artemisia capillaris, 15g of Solidago asiatica, 15g of Duckweed, 15g of Zedoariae rhizome, and 10g of Equisetum arvense.

[0019] Add 8 times and 6 times the amount of drinking water to the above drugs, extract for 1.5 hours and 1 hour, and filter. Concentrate the juice to a relative density of 1.2 (80°C), cool, add 2 times the amount of 70% ethanol, stir evenly, and after settling for 48 hours, take the supernatant and continue to concentrate to a relative density of 1.34-1.36 (59-61°C), pass through an 80-mesh sieve to obtain an extract, and mix with 0.8 times dextrin. After drying, crush, pass through a 60-mesh sieve, granulate, dry again, sieve, granulate, and mix to obtain granules.

[0020] Example 2 Antibacterial effects of the Chinese medicine composition and its disassembled prescriptions on Candida auris and Candida albicans

[0021] Experimental Materials:

[0022] RPMI 1640 complete medium: 89% RPMI 1640 basal medium + 10% FBS + 1% penicillin-streptomycin solution (100X).

[0023] Experimental strains: Candida auris (strain BJCA001) and Candida albicans (strain SC5314) were grown freely in YPD agar medium at 30°C for 48 h. To prepare the bacterial suspension, a single clone of cells was mixed with 50 ml sterile test tubes containing 10 ml YPD broth medium and cultured with shaking at 200 rpm and 30°C for 14-16 h. The cells were counted and used.

[0024] YPD agar medium: Sangon Biotech, CCM0110, China.

[0025] YPD broth medium: Sangon Biotech, CCM0210, China.

[0026] Colony counter: Hangzhou Xunshu Technology, Super G9, China.

[0027] Consumables: 96-well plates and bacterial culture plates.

[0028] Chenlanhua Granules: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Zedoaria 15g, Equisetum arvense 10g

[0029] Recipe 1: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Zedoaria 15g

[0030] Recipe 2: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Equisetum arvense 10g

[0031] Recipe 3: Artemisia capillaris 15g, Solidago asiatica 15g, Zedoariae 15g, Equisetum arvense 10g

[0032] Recipe 4: Artemisia capillaris 15g, duckweed 15g, zelan 15g, horsetail 10g

[0033] Prescription 5: 15g of Solidago, 15g of Duckweed, 15g of Zedoaria, and 10g of Equisetum arvense.

[0034] Experimental methods:

[0035] Microdilution method: MIC determination was performed according to the modified CLSI procedure (M27, fourth edition). Briefly, 100 μl of the drug with a final concentration of 2 times was serially diluted in RPMI1640 medium in a 96-well culture plate and mixed with 100 μl of fungal culture, and the fungal cell concentration was adjusted to 1 × 10 3cells / ml. Incubate at 30°C and read the optical density after 48 h using a Spectra Max ID3 plate reader (Molecular Devices, MD, USA); MIC is defined as the first well where the OD600 value drops by more than 50% compared with drug-free cells. Before reading the OD600 at 48 h, mix carefully by pipetting, and take photos after incubating the eggs at 30°C for 48 h. All experiments were performed and repeated three times.

[0036] Experimental results:

[0037] As shown in Table 1, the sensitivity of Candida auris and Candida albicans to Chen Lanhua Granules was tested. Chen Lanhua Granules had an antibacterial effect, and the minimum inhibitory concentration (MIC) was 256 μg / ml and 128 μg / ml, respectively, which was significantly better than the split prescription.

[0038] Table 1 Minimum inhibitory concentration

[0039]

[0040] Example 3 Effects of Chinese herbal medicine composition and its separate prescriptions on the survival rate of diabetic mice with Candida auris infection

[0041] Experimental Materials:

[0042] Experimental animals: C57BL / 6J male mice, 4-6 weeks old

[0043] Experimental drugs:

[0044] Chenlanhua Granules: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Zedoaria 15g, Equisetum arvense 10g

[0045] Recipe 1: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Zedoaria 15g

[0046] Recipe 2: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Equisetum arvense 10g

[0047] Recipe 3: Artemisia capillaris 15g, Solidago asiatica 15g, Zedoariae 15g, Equisetum arvense 10g

[0048] Recipe 4: Artemisia capillaris 15g, duckweed 15g, zelan 15g, horsetail 10g

[0049] Recipe 5: Goldenrod 15g, Duckweed 15g, Ze Lan 15g, Equisetum 10g

[0050] Experimental strain: Candida auris (BJCA001)

[0051] Experimental reagents and consumables: electronic balance, Sartorius, GL323-1SCN, China. Blood glucose meter, Aihao, CareSens C, China. Blood glucose test strips, Aihao, SB05LP54B1, China. Small animal anesthesia machine, Yuyan ABS, China. Streptozotocin, Sigma, S0130, USA. Sodium citrate, Solebol, C1013, China. Diabetes model feed, 74.5% mouse maintenance feed + 10% cooked lard + 10% sucrose + 5% egg yolk powder + 0.5% cholesterol, Jiangsu Collaborative Pharmaceutical Bioengineering.

[0052] Experimental methods:

[0053] After 1 week of adaptive feeding, C57BL / 6J mice were randomly divided into normal diet and high-fat diet groups. The high-fat diet group was fed a high-fat diet (HFD) for 4 consecutive weeks, while the normal diet group was maintained on a standard diet. The mice fed HFD were injected with STZ (100 mg / kg) twice for 3 consecutive days to establish a diabetic model. The diabetes combined with Candida auris infection group (DI-M, 1*10 8 CFU / mouse), Chenlanhua granules group (DI+CLH-M, 1.04g / kg), split prescription group 1 (DI+CF-1, 1.04g / kg), split prescription group 2 (DI+CF-2, 1.04g / kg), split prescription group 3 (DI+CF-3, 1.04g / kg), split prescription group 4 (DI+CF-4, 1.04g / kg), split prescription group 5 (DI+CF-5, 1.04g / kg). The mouse systemic infection model was performed by tail vein injection, and Chenlanhua granules were administered by oral gavage for 7 consecutive days to observe the survival of mice within 40 days.

[0054] Experimental results:

[0055] As shown in Table 2, the survival of mice after systemic infection with Candida auris was investigated and found that the survival rate of the DI-M group was 30%, the survival rate of the DI+CLH-M group was 75%, the survival rate of the DI+CF-1 group was 42%, the survival rate of the DI+CF-2 group was 45%, the survival rate of the DI+CF-3 group was 44%, the survival rate of the DI+CF-4 group was 45%, and the survival rate of the DI+CF-5 group was 40%. This shows that after treatment with each prescription, the survival rate of mice with diabetes combined with Candida auris infection was improved, but it was not as good as Chen Lanhua Granules.

[0056] Table 2 Effects of each prescription on the survival rate of diabetic mice with Candida auris infection

[0057]

[0058] Example 4 Effects of Chinese medicine composition and its separate prescriptions on bacterial load in mice with diabetes and Candida auris infection

[0059] Experimental Materials:

[0060] Experimental animals: C57BL / 6J male mice, 4-6 weeks old.

[0061] Experimental drugs:

[0062] Chenlanhua Granules: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Zedoaria 15g, Equisetum arvense 10g

[0063] Recipe 1: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Zedoaria 15g

[0064] Recipe 2: Artemisia capillaris 15g, Solidago asiatica 15g, Duckweed 15g, Equisetum arvense 10g

[0065] Recipe 3: Artemisia capillaris 15g, Solidago asiatica 15g, Zedoariae 15g, Equisetum arvense 10g

[0066] Recipe 4: Artemisia capillaris 15g, duckweed 15g, zelan 15g, horsetail 10g

[0067] Recipe 5: Goldenrod 15g, Duckweed 15g, Ze Lan 15g, Equisetum 10g

[0068] Experimental strain: Candida auris (BJCA001)

[0069] Experimental reagents and consumables: Electronic balance, Sartorius, GL323-1SCN, China. Blood glucose meter, Aihao, CareSens C, China. Blood glucose test strips, Aihao, SB05LP54B1, China. Small animal anesthesia machine, Yuyan ABS, China. Streptozotocin, Sigma, S0130, USA. Sodium citrate, Solebao, C1013, China. Diabetes model feed, 74.5% mouse maintenance feed + 10% cooked lard + 10% sucrose + 5% egg yolk powder + 0.5% cholesterol, Jiangsu Collaborative Pharmaceutical Bioengineering. YPD agar medium, Sangon Biotech, CCM0110, China. PBS buffer powder, Biosharp, BL601A, China. 4% paraformaldehyde fixative, Biosharp, BL539A, China. Hematoxylin, Sigma, H-3136, USA. Eosin, Sigma, E4009, USA. Periodic acid-Schiff (PAS) staining kit, Beyotime, C0142S, China.

[0070] Experimental methods:

[0071] After 1 week of adaptive feeding, C57BL / 6J mice were randomly divided into normal diet and high-fat diet groups. The high-fat diet group was fed a high-fat diet (HFD) for 4 consecutive weeks, while the normal diet group was maintained on a standard diet. The mice fed HFD were injected with STZ (100 mg / kg) twice for 3 consecutive days to establish a diabetic model. The diabetes combined with Candida auris infection group (DI-M, 1*10 8 CFU / mouse), Chenlanhua granules group (DI+CLH-M, 1.04g / kg), split prescription group 1 (DI+CF-1, 1.04g / kg), split prescription group 2 (DI+CF-2, 1.04g / kg), split prescription group 3 (DI+CF-3, 1.04g / kg), split prescription group 4 (DI+CF-4, 1.04g / kg), split prescription group 5 (DI+CF-5, 1.04g / kg). The mouse systemic infection model was performed by tail vein injection, and Chenlanhua granules were administered by oral gavage, and samples were collected after 7 consecutive days of administration.

[0072] The calculation formula for the bacterial load is: total bacterial load in the organ (CFU / mg) = number of bacteria on the bacterial plate / organ mass*dilution multiple.

[0073] Statistical methods: At least three independent samples were used for repeated experiments, and the results were expressed as mean ± standard error (x ± SEM), and statistical analysis was performed using GraphPad 9.3 statistical software. The t-test was used for comparison between two groups, and one-way analysis of variance or two-way analysis of variance was used for comparison between multiple groups. When the p value was less than 0.05, the difference was considered statistically significant.

[0074] Experimental results:

[0075] As shown in Table 3, compared with the diabetes combined with Candida auris infection group, after oral administration of Chen Lanhua granules, the fungal load in the kidneys and spleens of mice was significantly reduced, with statistical significance. Although each of the separated prescription groups reduced the fungal load in the kidneys and spleens of mice, it was not as good as the Chen Lanhua granules group. It can be seen that Chen Lanhua granules are better than each of the separated prescriptions in reducing the load of Candida auris.

[0076] Table 3. Effects of the separated prescriptions on bacterial load in mice with diabetes and Candida auris infection

[0077]

[0078] Note: *, compared with the normal group, P < 0.05; # , compared with the model group, P < 0.05; Δ, compared with the Chenlanhua granule group, P < 0.05.

[0079] Example 5 Comparative study of Chinese medicine composition and existing antifungal drugs

[0080] Clinical Data and Methods

[0081] Study subjects:

[0082] All patients visiting the outpatient and inpatient departments of the Department of Vascular Surgery at Shanghai Hospital of Integrated Traditional Chinese and Western Medicine with fungal infections.

[0083] Exclusion criteria:

[0084] ① Patients with severe liver and kidney dysfunction, systemic infectious diseases, malignant tumors and mental illness; ② Patients with severe allergic constitution; ③ Pregnant or lactating women; ④ Patients who are allergic to the test drug or its components.

[0085] Ethical Review:

[0086] This study has been reviewed by the Ethics Committee of Shanghai Hospital of Integrated Traditional Chinese and Western Medicine and has obtained the ethics approval ChiCTR2100052800.

[0087] Drug treatment:

[0088] Patients with fungal infection in the vascular disease department were given fluconazole, amphotericin B or caspofungin for symptomatic treatment according to the diagnosis and treatment standards; the Chinese medicine composition group (the Chinese medicine composition was prepared according to Example 1) was given the Chinese medicine composition, fluconazole + Chinese medicine composition, amphotericin B + Chinese medicine composition or caspofungin + Chinese medicine composition according to the diagnosis and treatment standards of the vascular disease department. Both groups were treated for 2 weeks, and renal function was rechecked. For patients with recurrent infection, the original plan was continued for treatment.

[0089] Observation indicators:

[0090] The effectiveness of drug intervention, the effect on renal function and the drug resistance after recurrent infection were observed in each group.

[0091] As shown in Table 4, compared with the fluconazole group alone, the addition of the Chinese medicine composition significantly reduced the emergence of drug resistance (P < 0.05). As shown in Table 5, compared with amphotericin B alone, the addition of Chinese medicine treatment significantly reduced the damage of amphotericin B to the kidney (P < 0.05). As shown in Table 6, compared with caspofungin alone, the addition of the Chinese medicine composition significantly improved its effectiveness (P < 0.05).

[0092] Table 4 Comparison of fluconazole resistance (example)

[0093]

[0094] Table 5 Comparison of renal injury caused by amphotericin B (examples)

[0095]

[0096] Table 6 Comparison of the effectiveness of caspofungin

[0097]

[0098] The various aspects of the present invention have been described above. However, it should be understood that, without departing from the spirit of the present invention, equivalent changes and modifications may be made by those skilled in the art, and the changes and modifications also fall within the scope of the appended claims of the present application.

Claims

1. Use of a Chinese medicine composition in the preparation of a drug for preventing and treating diabetes combined with Candida infection, the Chinese medicine composition being prepared from the following raw materials in parts by weight: 8-25 parts of Artemisia capillaris, 8-25 parts of Solidago asiatica, 8-25 parts of Duckweed, 8-25 parts of Zedoariae, and 6-15 parts of Equisetum arvense, wherein the Candida is Candida auris or Candida albicans.

2. The use according to claim 1, wherein the traditional Chinese medicine composition is made of the following raw materials in parts by weight: 15 parts of Artemisia capillaris, 15 parts of Solidago asiatica, 15 parts of Duckweed, 15 parts of Zedoariae rhizome, and 10 parts of Equisetum arvense.

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