Use of traditional chinese medicine in preparing drug for preventing and treating nephritis

Abstract

Use of a traditional Chinese medicine in preparing a drug for preventing and treating nephritis. The traditional Chinese medicine is prepared from Taraxacum officinale, Polygonum cuspidatum, Sonchus oleroceus L., and Herba Scutellariae barbatae. Danio rerio is adopted for modeling. Animal experiments prove that the traditional Chinese medicine can be used for treating nephritis caused by aristolochic acid, protecting kidney functions and / or ameliorating renal dysfunction edema, and specifically, promoting glomerular filtration.

Description

Application of traditional Chinese medicine in the preparation of medicines for preventing and treating nephritis

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on December 25, 2023, with application number 202311798771.5 and invention name “Application of Traditional Chinese Medicine in the Preparation of Drugs for the Prevention and Treatment of Nephritis”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention relates to the technical field of traditional Chinese medicines, and in particular to the application of traditional Chinese medicines in the preparation of medicines for preventing and treating nephritis. Background Art

[0003] Chronic glomerulonephritis is currently the leading cause of chronic renal failure in my country. The glomerular filtration rate (GFR) accurately reflects the filtration function of the kidneys and is an immune inflammatory response that occurs in the glomeruli. Clinically, there are many causes of nephritis. When patients are infected with bacteria or viruses, or due to allergies, autoimmune disorders, or medications, immune complexes can form. After the immune complexes are deposited in the glomeruli, the glomerular basement membrane may thicken, and glomerular mesangial cells may proliferate, the mesangial matrix may increase, and capillary endothelial cells may proliferate.

[0004] Currently, glucocorticoids are commonly used to treat nephritis. However, glucocorticoids only suppress immunity and provide anti-inflammatory measures, failing to fully eliminate the multiple pathogenic factors that damage glomerular resident cells and thereby completely repair them. Furthermore, glucocorticoids have significant toxic side effects. While long-term glucocorticoid use may cure the disease, it also significantly reduces immunity. For example, Wang Li et al. conducted a statistical analysis of 199 cases of adverse reactions to dexamethasone reported in various domestic journals. The results showed that adverse reactions to dexamethasone include systemic damage, damage to the central and peripheral nervous systems, psychiatric symptoms, damage to the musculoskeletal system, damage to the skin and its appendages, visual impairment, general damage to the cardiovascular system, digestive system damage, respiratory system damage, endocrine disorders, reproductive system damage, metabolic and nutritional disorders, hepatobiliary system damage, platelet abnormalities, and coagulation abnormalities. Among these, anaphylactic shock is a key concern (see "Analysis of 199 Cases of Adverse Reactions to Dexamethasone" by Wang Li et al.).

[0005] Under the influence of side effects, patients' conditions are prone to recurring and difficult to cure. Traditional Chinese medicine is based on dialectical treatment. While treating nephritis, it can also protect kidney function and delay the progression of chronic renal failure. Traditional Chinese medicine treatment methods do not cause too much pain to patients, do not have too many side effects, are very convenient to use, and can fundamentally solve the problem. Therefore, it is necessary to find a drug that can treat nephritis without the above side effects or with reduced side effects. Generally, Western medicine has relatively large side effects, a long development cycle, and high development costs. Finding a traditional Chinese medicine that can prevent and treat nephritis is a good solution. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide the use of traditional Chinese medicine in the preparation of drugs for preventing and treating nephritis. While using traditional Chinese medicine to treat nephritis, it can further protect the kidneys, delay the progression of chronic renal failure, and fundamentally regulate the body.

[0007] The present invention provides application of traditional Chinese medicine in preparing medicine for preventing and treating nephritis; wherein the traditional Chinese medicine is prepared from dandelion, giant knotweed, patrinia chinensis and scutellaria barbata.

[0008] The present invention provides a Chinese patent medicine for treating nephritis. Experimental studies have shown that it has the effect of protecting kidney function, specifically by promoting glomerular filtration. The medicine can delay the progression of chronic renal failure and fundamentally regulate the body.

[0009] In the present invention, the nephritis is chronic glomerulonephritis.

[0010] In the embodiment of the present invention, the nephritis is nephritis caused by drug damage.

[0011] In some embodiments, the drug is aristolochic acid.

[0012] In the present invention, the prevention and treatment of nephritis includes protecting renal function. In some embodiments, protecting renal function includes promoting glomerular filtration.

[0013] In the present invention, the prevention and treatment of nephritis includes improving edema caused by renal dysfunction.

[0014] In the present invention, the term "prevention" includes "prevention" and / or "treatment," wherein "treatment" refers to the process of eliminating the cause or focus of a disease with a living human or animal body as the direct subject. Beneficial or desired clinical outcomes of the treatment include, but are not limited to, alleviation of symptoms, attenuation of disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, and remission (whether partial or complete), whether detectable or undetectable.

[0015] In the present invention, the objects of prevention and treatment include objects already suffering from a disease or illness as well as objects susceptible to the disease or illness. When referring to terms such as alleviation, alleviation, reduction, alleviation, and relief, the meanings thereof also include situations such as elimination, disappearance, and non-occurrence.

[0016] In the present invention, the subject of treatment refers to an organism receiving treatment with the Chinese medicine of the present invention, including mammals or non-mammals receiving treatment for a disease or condition. Exemplary mammals include bovines, equines, ovines, porcines, canines, felines, rodents, and primates. Preferably, the mammal is a human. In the Examples, the efficacy of the treatment is demonstrated using a non-mammalian animal, specifically a zebrafish.

[0017] In the present invention, the effective dose for preventing and treating nephritis is 0.061 μL / mL to 0.244 μL / mL, for example, 0.061 μL / mL, 0.122 μL / mL or 0.244 μL / mL, equivalent to a human dose of 21.65 mL to 86.62 mL.

[0018] As previously described, an effective dose refers to an amount of a therapeutic agent that, when administered to a cell, tissue, or subject, is effective in preventing or ameliorating the symptoms of a disease or the progression of that disease. "Effective amount" also refers to an amount of a compound sufficient to alleviate symptoms, e.g., to treat, cure, prevent, or alleviate a related medical condition, or to increase the rate of treatment, cure, prevention, or alleviation of such a condition. When an active ingredient is administered alone to a subject, a therapeutically effective dose refers to that ingredient alone. When a combination is administered, a therapeutically effective dose refers to the combined amounts of the active ingredients that produce a therapeutic effect, whether administered in combination, sequentially, or simultaneously.

[0019] In the present invention, the traditional Chinese medicine is prepared from the following raw materials in parts by mass: 372 parts of dandelion, 372 parts of polygonum cuspidatum, 372 parts of patrinia ulmoides and 186 parts of scutellaria barbata.

[0020] In some embodiments, the traditional Chinese medicine further comprises a pharmaceutically acceptable carrier or excipient. Specifically, the pharmaceutical composition is a solid preparation, a mixture, an injection, an external preparation, a spray, a liquid preparation, or a compound preparation.

[0021] In some embodiments, the traditional Chinese medicine is prepared by decoction.

[0022] In some specific embodiments, the preparation method of the traditional Chinese medicine comprises extracting the raw materials through water decoction, filtering the extract and then centrifuging it, adding stevia and ethylparaben to the supernatant, and boiling it to obtain the traditional Chinese medicine.

[0023] The decoction is performed twice, with 5 to 15 times the weight of the medicinal material added in water for the first time and decocted for 2 hours, and 4 to 10 times the weight of the medicinal material added in water for the second time and decocted for 1 hour. Preferably, 8 times the weight of water is added in the first time and 5 times the weight of water is added in the second time.

[0024] The present invention also provides a method for preventing and treating nephritis, which comprises administering the above-mentioned traditional Chinese medicine.

[0025] In the present invention, the administration of the Chinese medicine includes, but is not limited to, oral administration, spray inhalation, rectal administration, nasal administration, buccal administration, vaginal administration, topical administration, parenteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or administration via an explanted reservoir. Among them, oral, intraperitoneal or intravenous administration is preferred.

[0026] In the experimental scheme of the present invention, zebrafish are used for modeling, and animal experiments are used to prove that the traditional Chinese medicine as described above can treat nephritis and has a protective effect on renal function, specifically by promoting glomerular filtration. In addition, the traditional Chinese medicine has good safety and will not produce serious side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 shows a typical graph of whole-body fluorescence intensity of zebrafish after sample treatment. DETAILED DESCRIPTION

[0028] The present invention provides the application of Chinese medicine in the preparation of medicines for preventing and treating nephritis. Those skilled in the art can refer to the contents of this article and appropriately improve the process parameters to achieve the desired effect. It should be noted that all similar replacements and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0029] The present invention provides the use of traditional Chinese medicine in preventing and treating nephritis. The traditional Chinese medicine used in the present invention is made of dandelion, polygonum cuspidatum, northern patrinia and Scutellaria barbata, which has the effect of protecting renal function. Renal function can be divided into glomerular function and tubular function. Glomerular function is mainly the determination of glomerular filtration rate, which can be evaluated by testing blood urea nitrogen and creatinine levels. Glomerular function can also be accurately evaluated by calculating endogenous creatinine clearance. Tubular function refers to the reabsorption and excretion function of the renal tubules. Urinary NAG is often used clinically to reflect whether there is renal tubular damage. In addition, renal tubular function can also be evaluated by urine β2 microglobulin, urine retinol binding protein, urine specific gravity, urine osmotic pressure, etc. In the present invention, by detecting the glomerular filtration rate, it is proved that the traditional Chinese medicine has the effect of protecting renal function, especially in the renal dysfunction edema model. Thus, it is proved that the traditional Chinese medicine has the effect of preventing and treating nephritis.

[0030] The traditional Chinese medicine obtained by screening of the present invention is safer and has fewer side effects than the commonly used drug dexamethasone in treating nephritis. It will not produce side effects such as systemic damage, anaphylactic shock, damage to the central and peripheral nervous systems, mental symptoms, damage to the musculoskeletal system, damage to the skin and its appendages, visual damage, general damage to the cardiovascular system, damage to the digestive system, damage to the respiratory system, endocrine disorders, damage to the reproductive system, metabolic and nutritional disorders, damage to the hepatobiliary system, platelet abnormalities or coagulation abnormalities.

[0031] The examples of the present invention only record a case of a Chinese medicine composition as a test product. In addition, many attempts were made during the research and development process, such as using different Chinese medicinal materials or Chinese medicine compositions, using different extraction methods, etc. However, it was found in these experiments that whether it was a Chinese medicinal material such as dandelion alone for the experiment, or using other Chinese patent medicines, or a composition formed with dandelion as one of the components, its protective effect on renal function was not as good as that of the Chinese medicine prepared from dandelion, knotweed, northern patrinia and bark lotus. The specific experimental results are not repeated here. The results show that dandelion, knotweed, northern patrinia and bark lotus cooperate with each other and are indispensable. Under the appropriate ratio, they can produce a synergistic effect, thereby playing a more significant role in protecting renal function.

[0032] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0033] The present invention will be further described below in conjunction with the embodiments:

[0034] Example: Renal function protection effect

[0035] 1. Testing materials

[0036] 1.1. Sample preparation information

[0037] Chinese medicine decoction: The Chinese medicine involved in the embodiment of the present invention is prepared from dandelion (main drug), polygonum cuspidatum (minister drug), northern patrinia (adjuvant drug) and bark stalk (adjuvant drug). Its preparation method comprises: taking dandelion 372g, polygonum cuspidatum 372g, northern patrinia 372g, and bark stalk 186g, adding water and decocting twice, the first time adding 8 times the water and decocting for 2 hours, the second time adding 5 times the water and decocting for 1 hour, filtering the decoction, concentrating the filtrate under reduced pressure, combining the concentrate, centrifuging, filtering, adding stevioside 1.5g and ethylparaben 0.5g, heating to boiling, and making 1000ml.

[0038] For the test product, the solvent was 1×E3 culture medium, and the above decoction was mixed with E3 culture medium.

[0039] Experimental Animals

[0040] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to 1 L of reverse osmosis water, conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3). They were bred and provided by our company's fish farming center. The experimental animal use license number is: SYXK(Zhejiang)2022-0004. The breeding and management of zebrafish complies with the requirements of the international AAALAC accreditation (certification number: 001458) and the IACUC ethics review number is: IACUC-2023-7687-01.

[0041] 1.3. Instruments, consumables, and reagents

[0042] A dissecting microscope (SZX7, OLYMPUS, Japan); a CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); a motorized focus continuous zoom fluorescence microscope (AZ100, Nikon, Japan); a microinjector (IM300, Narishige, Japan); a needle puller (PC-10, Narishige, Japan); a precision electronic balance (CP214, OHAUS, USA); and a 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China).

[0043] Methylcellulose (Batch No. C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China); dimethyl sulfoxide (DMSO, Batch No. BCCD8942, Sigma, Switzerland); aristolochic acid (Batch No. E2015060, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Dextrantetramethylrhodamine (Batch No. 923446, Molecular Probes, USA).

[0044] 2. Detection Method

[0045] 2.1. Determination of maximum detectable concentration (MTC)

[0046] Wild-type AB zebrafish, 2 days post-fertilization (dpf), were randomly selected and plated in 6-well plates. Thirty zebrafish were treated in each well (experimental group). Samples (concentrations shown in Table 1-1) were administered in water. A normal control group and a model control group were also established, with a volume of 3 mL per well. Except for the normal control group, all other experimental groups were administered aristolochic acid in water to establish a zebrafish model of renal dysfunction and edema. After treatment at 28°C for 2 days, the MTC of the samples in the model zebrafish was measured.

[0047] 2.2. Kidney function protection effect (glomerular filtration)

[0048] Wild-type AB strain zebrafish (2 dpf) were randomly selected and plated in 6-well plates. Thirty zebrafish were treated in each well (experimental group). Samples (concentrations shown in Tables 1-2) were administered in water. A normal control group and a model control group were also established. Each well held 3 mL of water. All experimental groups, except the normal control group, were administered aristolochic acid in water to establish a zebrafish model of renal edema. After one day of treatment at 28°C, each experimental group received an intravenous injection of a fluorescent marker (dextran tetramethylrhodamine). Treatment continued at 28°C for another day. Ten zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. Data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. Whole-body fluorescence intensity was analyzed, and statistical analysis of this indicator was used to evaluate the renal function protective efficacy of the samples (glomerular filtration). Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. P < 0.05 indicated statistical significance.

[0049] 3. Test results

[0050] 3.1.MTC

[0051] Under the experimental conditions, the MTC of the test product on renal function protection was 0.244 μL / mL. See Table 1-1 for details.

[0052] Table 1-1. Results of the experimental study on the concentration of the sample for the protection of renal function (n=30)

[0053] 3.2. Evaluation of renal function protection efficacy (glomerular filtration)

[0054] Under the experimental conditions, the test product demonstrated renal protective effects, specifically by promoting glomerular filtration. See Table 1-2 and Figure 1 for details.

[0055] Table 1-2. Results of the renal function protection efficacy evaluation (glomerular filtration) experiment of the samples (n=10) Compared with the model control group, *p<0.05, ***p<0.001

[0056] The above are only preferred embodiments of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of traditional Chinese medicine in the preparation of drugs for preventing and treating nephritis; The traditional Chinese medicine is made from dandelion, polygonum cuspidatum, patrinia scabiosaefolia and scutellaria barbata.

2. The application according to claim 1, wherein The nephritis is chronic glomerulonephritis.

3. The application according to claim 2, wherein The nephritis is nephritis caused by drug injury.

4. The application according to claim 3, wherein The drug is aristolochic acid.

5. The application according to claim 1, characterized in that, The prevention and treatment of nephritis includes protecting renal function and / or improving renal dysfunction edema.

6. The application according to claim 5, wherein The protection of renal function includes promoting glomerular filtration.

7. The application according to claim 1, wherein The effective dose for preventing and treating nephritis is 0.061 μL / mL to 0.244 μL / mL, which is equivalent to the human dose of 21.65 mL to 86.62 mL.

8. The application according to any one of claims 1 to 7, characterized in that, The traditional Chinese medicine is prepared from the following raw materials by mass: 372 parts of dandelion, 372 parts of polygonum cuspidatum, 372 parts of patrinia scabiosaefolia and 186 parts of scutellaria barbata.

9. The application according to claim 8, characterized in that, The preparation method of the traditional Chinese medicine includes decocting the raw materials with water, filtering the extract and then centrifuging, adding steviol glycoside and ethylparaben to the supernatant, and boiling to obtain the traditional Chinese medicine.

10. A method for preventing and treating nephritis, characterized in that, Including administering the traditional Chinese medicine; The traditional Chinese medicine is prepared from the following raw materials by mass: 372 parts of dandelion, 372 parts of polygonum cuspidatum, 372 parts of patrinia scabiosaefolia and 186 parts of scutellaria barbata.

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