Medicine for treating acute glomerulonephritis

Abstract

The invention discloses a medicine for treating acute glomerulonephritis, and belongs to the technical field of medicines. The medicine for treating acute glomerulonephritis comprises at least one of asiatic acid, N-acetyltryptophan and catechin gallate as effective components. The effective components in the medicine are all natural active components extracted from plants, and are safe and free of toxic and side effects. When the medicine is used for intragastric administration of mice with acute glomerulonephritis, the contents of urine protein, urea nitrogen, occult blood, microscopic examination red blood cells and microscopic examination white blood cells of the mice can be remarkably reduced, and the ingestion of the mice with acute glomerulonephritis can be improved, so that the weight is increased, and the effect is superior to that of nephritis rehabilitation tablets. Therefore, the medicine disclosed by the invention has a great application prospect in the treatment of acute glomerulonephritis.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceuticals, and particularly relates to a drug for treating acute glomerulonephritis. Background Art

[0002] Acute glomerulonephritis is a group of primary glomerulonephritides with the main clinical manifestations of acute nephritis syndrome, which is characterized by: acute onset, hematuria, proteinuria, edema and hypertension, and may be accompanied by transient azotemia, with a tendency to self-heal. It is common after streptococcal infection, while other bacterial, viral and parasitic infections can also cause it.

[0003] Currently, the drugs for treating acute glomerulonephritis include Chinese patent medicines, antibiotics, diuretics, immunosuppressants, glucocorticoids, antihypertensive drugs, etc. Among them, Chinese patent medicines such as Shenyan Kangfu Tablets have a stimulating effect on the gastrointestinal mucosa. After taking them, some patients may have gastrointestinal discomfort symptoms such as nausea, vomiting, and diarrhea, and a few patients may also have allergic reactions after taking them, such as rashes, itching, and difficulty breathing. If taken in large amounts for a long time, it may also cause certain effects on liver and kidney functions, resulting in abnormal liver and kidney functions. Antibiotic drugs such as penicillin and amoxicillin have the risk of causing allergic reactions, and abuse may lead to drug resistance and are ineffective for non-infectious AGN (such as IgA nephropathy). Long-term use of diuretic drugs such as furosemide can cause electrolyte disorders (hypokalemia, hyponatremia) and may exacerbate the azotemia of patients with renal insufficiency. Taking immunosuppressants such as cyclophosphamide and mycophenolate mofetil can significantly increase the risk of infection (such as pneumonia, sepsis). For example, cyclophosphamide may cause bone marrow suppression and hemorrhagic cystitis, and mycophenolate mofetil may cause gastrointestinal reactions (diarrhea, nausea). Long-term use of glucocorticoids (such as prednisone) can lead to osteoporosis, elevated blood sugar, obesity, and sudden withdrawal may trigger adrenal crisis, masking the symptoms of infection and delaying treatment. ACEI / ARB, such as enalapril and losartan, may cause dry cough (due to bradykinin accumulation), the risk of hyperkalemia (especially in patients with renal insufficiency), and need to be used with caution in severe renal failure, and may increase serum creatinine.

[0004] In summary, the currently used drugs for treating acute glomerulonephritis all have problems such as large side effects and poor treatment effects. Therefore, there is an urgent need to provide a preparation with good treatment effect and no toxic side effects for acute glomerulonephritis. Summary of the Invention

[0005] Aiming at the problems existing in the prior art, the purpose of the present invention is to provide a drug for treating acute glomerulonephritis.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0007] A drug for treating acute glomerulonephritis, the active ingredient being at least one of asiatic acid, N-acetyltryptophan, and catechin gallate.

[0008] On the basis of the above scheme, the mass ratio of asiatic acid, N-acetyltryptophan, and catechin gallate is: 0-6:0-3:0-3.

[0009] On the basis of the above scheme, the active ingredient of the drug for treating acute glomerulonephritis is asiatic acid, N-acetyltryptophan, and catechin gallate.

[0010] On the basis of the above scheme, the mass ratio of N-acetyltryptophan and catechin gallate is 1:1.

[0011] On the basis of the above scheme, the mass ratio of asiatic acid, N-acetyltryptophan, and catechin gallate is 2:5:5-10:1:1.

[0012] On the basis of the above scheme, the mass ratio of asiatic acid, N-acetyltryptophan, and catechin gallate is 2:1:1.

[0013] On the basis of the above scheme, the drug for treating acute glomerulonephritis further comprises pharmaceutically acceptable excipients.

[0014] On the basis of the above scheme, the drug dosage form is at least one of tablets, capsules, granules, pills, and oral liquids.

[0015] Advantages of the technical solution of the present invention:

[0016] For the drug for treating acute glomerulonephritis of the present invention, the active ingredients asiatic acid, N-acetyltryptophan, and catechin gallate in the drug are all natural active ingredients extracted from plants, and are safe and non-toxic with no side effects.

[0017] When the drug of the present invention is used for intragastric administration to mice with acute glomerulonephritis, it can significantly reduce the contents of urinary protein, blood urea nitrogen, urinary occult blood, microscopic examination of red blood cells, and microscopic examination of white blood cells in mice, and can improve the food intake of mice with acute glomerulonephritis, thereby increasing weight, and the effect is better than that of Shenyan Kangfu Tablets. Therefore, the drug of the present invention has great application prospects in the treatment of acute glomerulonephritis. Description of the Drawings

[0018] Figure 1 Cell survival rate after incubation of Examples 1-7 with J774A.1 mouse macrophages. Detailed Embodiments

[0019] The terms used in the present invention generally have the meanings commonly understood by those of ordinary skill in the art, unless otherwise specified. The present invention will be described in further detail below with reference to specific embodiments and data. The following embodiments are only for illustrative purposes of the present invention and do not limit the scope of the present invention in any way.

[0020] The experimental methods in the following embodiments are all conventional methods, unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in the art or according to the product specifications. The test materials, reagents, drugs, etc. used in the following embodiments can be obtained through general channels, unless otherwise specified.

[0021] In the following embodiments,

[0022] Asiatic acid (98%) CAS No.: 464-92-6;

[0023] Catechin gallate (≥98%) CAS No.: 130405-40-2;

[0024] N-acetyltryptophan (98%) CAS No.: 1218-34-4;

[0025] BSA is bovine serum albumin, and SEB is staphylococcal enterotoxin B, which are drugs for modeling.

[0026] The drug specification of Shenyan Kangfu Tablets is 0.3g * 72 tablets, and the national drug approval number is Z10940029.

[0027] In the following embodiments,

[0028] The mice were placed in metabolic cages, and urine was collected for 24 hours. During this period, feed and free drinking water were provided to avoid interference from stress-induced urination. After urine collection, it was immediately stored at 4°C. The urine was centrifuged and the precipitate was taken, smeared and observed under a microscope for the number and morphology of red blood cells and white blood cells. Counting was performed per high-power field (HPF), and the normal reference value was: white blood cells < 5 / HPF, red blood cells < 3 / HPF. Morphological abnormalities (such as deformed red blood cells) indicate glomerular hematuria, and comprehensive judgment needs to be combined with urine protein quantification.

[0029] An URIT-14G automatic urine analyzer was used, and the photoelectric colorimetric method and multi-wavelength (480 - 660nm) reflection light detection technology were adopted to detect the urine protein content, urine occult blood content, etc.

[0030] Mouse blood samples were collected, the supernatant was taken after centrifugation, and a urea nitrogen (BUN) detection kit (urease-glutamate dehydrogenase method) was used to detect the urea nitrogen content in the mouse serum.

[0031] Example 1

[0032] A drug for treating acute glomerulonephritis is composed of N-acetyltryptophan and epigallocatechin gallate in a mass ratio of 1:1.

[0033] Example 2

[0034] A drug for treating acute glomerulonephritis is composed of asiatic acid, N-acetyltryptophan and epigallocatechin gallate in a mass ratio of 2:5:5.

[0035] Example 3

[0036] A drug for treating acute glomerulonephritis is composed of asiatic acid, N-acetyltryptophan and epigallocatechin gallate in a mass ratio of 1:1:1.

[0037] Example 4

[0038] A drug for treating acute glomerulonephritis is composed of asiatic acid, N-acetyltryptophan and epigallocatechin gallate in a mass ratio of 2:1:1.

[0039] Example 5

[0040] A drug for treating acute glomerulonephritis is composed of asiatic acid, N-acetyltryptophan and epigallocatechin gallate in a mass ratio of 4:1:1.

[0041] Example 6

[0042] A drug for treating acute glomerulonephritis is composed of asiatic acid, N-acetyltryptophan and epigallocatechin gallate in a mass ratio of 10:1:1.

[0043] Example 7

[0044] A drug for treating acute glomerulonephritis, the active ingredient of which is asiatic acid.

[0045] Therapeutic effect of the drug for treating acute glomerulonephritis on acute glomerulonephritis in mice

[0046] Model establishment of acute glomerulonephritis in mice: KM mice (SPF grade), weighing 20 g, with half male and half female, a total of 72 mice. After one week of adaptive feeding, in the model group, in addition to free drinking water and diet, the mice were given intragastric administration of BSA (200 mg / kg) every other day for the first 5 weeks; from the 6th week, BSA (20 mg / kg, 500 mg BSA dissolved in 500 mL of sterile physiological saline) was injected via the tail vein once a day for 3 consecutive days; from the 8th week, SEB (0.5 mg / kg, 1 mg SEB dissolved in 25 mL of sterile physiological saline) was injected via the tail vein once a week for 3 consecutive weeks.

[0047] The above-mentioned modeled mice were divided into a model group, a drug control group, and an experimental group; among them, 8 mice in the model group were normally raised. 8 mice in the drug control group were gavaged with Shenyan Kangfu Tablets, and the gavage dose for a 20g mouse was 0.6mg each time, and they were gavaged once at 8:00 am, 12:00 noon, and 5:00 pm, and they were given normal drinking water during other feeding times. The experimental group was gavaged with drugs according to the ratios of Examples 1-7, with 8 mice for each drug, and the gavage dose for a 20g mouse was 0.6mg each time (Table 1), and they were gavaged once at 8:00 am, 12:00 noon, and 5:00 pm. They were given normal drinking water during other feeding times. The 8 mice in the normal feeding group refer to the mice before modeling without any treatment.

[0048] Table 1 Components of the drugs gavaged to the mice in the experimental group

[0049]

[0050] After 28 days of the experiment, the urine protein content of the mice in each group was detected, and the results are shown in Table 2.

[0051] Table 2 Changes in urine protein content (g / L)

[0052]

[0053] As can be seen from Table 2, compared with the model group, the urine protein content of the mice in the groups of Examples 1-7 all showed a significant decrease after 28 days of treatment, especially in the group of Example 4, the urine protein content of the mice was all 0 (not detected). It is worth mentioning that although the urine protein content of the mice in the Shenyan Kangfu Tablets group also showed a significant decrease, there were still 3 mice with a urine protein content of 0.15g / L (weak positive). Through the above analysis, it can be seen that the therapeutic effect of the drug in Example 4 is better than that of the Shenyan Kangfu Tablets treatment group.

[0054] After 28 days of the experiment, the blood urea nitrogen content of the mice in each group was detected, and the results are shown in Table 3.

[0055] Table 3 Changes in blood urea nitrogen content (mmol / L)

[0056]

[0057] Analysis of the data in Table 3 shows that in terms of reducing the blood urea nitrogen content of mice, the effect of Example 4 is also better than that of the Shenyan Kangfu Tablets group. Although the therapeutic effects of the other example groups are slightly worse than those of the Shenyan Kangfu Tablets group, compared with the model group, they also show a good therapeutic effect.

[0058] After 28 days of the experiment, the urine occult blood content of the mice in each group was detected, and the results are shown in Table 4.

[0059] Table 4 Changes in urine occult blood content (CELL / UL)

[0060]

[0061] By analyzing the changes in the occult blood content in the urine of the mice in Table 4, it was found that Examples 1-7 could all reduce the occult blood content in the urine of the mice. In particular, Examples 3 and 4, especially in the group of Example 4, the occult blood content in the urine of the mice reached 0 after 28 days of treatment. And the therapeutic effect of Example 3 was comparable to that of the group treated with Shenyan Kangfu Tablets.

[0062] After 28 days of the experiment, the content of microscopic examination of red blood cells in each group of mice was detected, and the results are shown in Table 5.

[0063] Table 5 Changes in the content of microscopic examination of red blood cells (RBC / HP)

[0064]

[0065] After analyzing the data in Table 5, it was found that compared with the model group, the content of microscopic examination of red blood cells in the mice in Examples 1-7 groups all showed different degrees of decline after 28 days of treatment. In particular, the content of microscopic examination of red blood cells in the mice in the group of Example 4 was not detected after 28 days of treatment.

[0066] After 28 days of the experiment, the content of microscopic examination of white blood cells in each group of mice was detected, and the results are shown in Table 6.

[0067] Table 6 Changes in the content of microscopic examination of white blood cells (WBC / HP)

[0068]

[0069] As can be seen from Table 6, compared with the model group, the content of microscopic examination of white blood cells in the mice in Examples 1-7 groups all showed a significant decline after 28 days of treatment. Among them, the therapeutic effects of Examples 3 and 5 were comparable to that of the group treated with Shenyan Kangfu Tablets. It is worth mentioning that the content of microscopic examination of white blood cells in the mice in the group of Example 4 was not detected after 28 days of treatment. It can be seen that in this regard, the therapeutic effect of Example 4 is better than that of the treatment group with Shenyan Kangfu Tablets.

[0070] After 28 days of the experiment, the body weights of each group of mice were weighed, and the results are shown in Table 7.

[0071] Table 7 Changes in the body weight of mice (g)

[0072]

[0073] As can be seen from Table 7, compared with the body weights of the mice in the model group, the body weights of the mice in each medicated group all increased, especially in Example 4. This result indicates that the drug provided in the present invention can improve the feeding of mice with acute glomerulonephritis, thereby increasing the weight.

[0074] In order to further confirm the therapeutic effect of the drug in Example 4, after 28 days of the experiment, the mice in the Example 4 group stopped taking the drug and were continuously fed for 14 days. The changes in relevant indicators of the mice were detected on the 14th day as follows:

[0075] Table 8 Changes in various indicators of mice after stopping the drug

[0076]

[0077] As can be seen from Table 8, on the 14th day after stopping the drug, the indicators of the mice in the Example 4 group all reached the indicators of healthy mice. Among them, the contents of urinary occult blood and urinary protein were both 0, and the contents of red blood cells and white blood cells detected by microscopy were not detected. And except for body weight, the other indicators tended to be stable. The increase in the body weight of the mice was because the provided drug could improve the feeding of mice with acute glomerulonephritis, thus increasing the weight. All in all, the data in Table 8 show that the drug in Example 4 has a curative effect on mice with acute glomerulonephritis.

[0078] Cytotoxicity test

[0079] The cytotoxicity of Examples 1-7 was determined by the cell thiazolyl blue (MTT) colorimetric method. Mouse macrophages J774A.1 were placed in a cell culture dish and incubated overnight at 37 °C and 5% CO2 for standby. The cells were washed with pre-warmed Hanks buffer (HNSS) at 37 °C. While setting up a blank group, the drugs corresponding to Examples 1-7 were added to each culture dish respectively, so that the final concentration of the drug in the culture dish was 0.03 mg / mL, and incubated at 37 °C for 4 h. Subsequently, 0.5 mg / mL MTT solution was added and incubated at 37 °C for 4 hours. After removing the excess MTT solution, an appropriate amount of dimethyl sulfoxide (DMSO) was added to form blue-violet crystals. An enzyme-linked immunosorbent assay (ELISA) reader was used to measure the absorbance at 560 nm and calculate the cell survival rate: Cell survival rate = OD value of the experimental group / OD value of the blank group × 100%.

[0080] The results are as Figure 1 shown. The survival rates of the cells treated with Examples 1-7 were all above 98.5%. According to the 70% cytotoxicity critical value recommended by ISO10993-5:2009 (Biological evaluation of medical devices - Part 5: Tests for in vitro cytotoxicity), it shows that the cytotoxicity of Examples 1-7 to J774A.1 cells can be almost ignored. This further proves that the natural plant components used in Examples 1-7 are safe for the human body.

[0081] The above are only the preferred embodiments of the present invention, and are not intended to limit the present invention in other forms. Any person skilled in the art may use the disclosed technical content to make changes or modifications into equivalent embodiments with equivalent changes. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the technical solution content of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims

1. A drug for treating acute glomerulonephritis, characterized in that, The active ingredient is at least one of asiatic acid, N-acetyltryptophan, and epigallocatechin gallate.

2. The drug for treating acute glomerulonephritis according to claim 1, characterized in that, The mass ratio of asiatic acid, N-acetyltryptophan, and epigallocatechin gallate is: 0-6:0-3:0-3.

3. The drug for treating acute glomerulonephritis according to claim 1, wherein The active ingredient is asiatic acid, N-acetyltryptophan, and epigallocatechin gallate.

4. The drug for treating acute glomerulonephritis according to claim 3, characterized in that, The mass ratio of N-acetyltryptophan to epigallocatechin gallate is 1:

1.

5. The drug for treating acute glomerulonephritis according to claim 4, characterized in that, The mass ratio of asiatic acid, N-acetyltryptophan, and epigallocatechin gallate is 2:5:5-10:1:

1.

6. The drug for treating acute glomerulonephritis according to claim 5, characterized in that, The mass ratio of asiatic acid, N-acetyltryptophan, and epigallocatechin gallate is 2:1:

1.

7. The drug for treating acute glomerulonephritis according to any one of claims 1-6, characterized in that, It further contains pharmaceutically acceptable excipients.

8. The drug for treating acute glomerulonephritis according to claim 7, characterized in that, The pharmaceutical dosage form is at least one of tablets, capsules, granules, pills, and oral liquids.

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