Application of Danshufu Fuyuan Formula in preparation of drugs for treating idiopathic membranous nephropathy

Abstract

The application belongs to the field of medicine, and particularly relates to application of a Danzhu Fuguan formula in preparation of a drug for treating idiopathic membranous nephropathy (IMN). The Danzhu Fuguan formula comprises the following raw materials in parts by weight: 25-35 parts of Danshen, 10-15 parts of Baizhu, 25-35 parts of Huangqi, 10-20 parts of Jinyinhua, 10-20 parts of Hujiang, 10-20 parts of Yujin, 10-15 parts of Fuling, 25-35 parts of Yiyiren and 10-15 parts of Qianghuo. Research shows that the Danzhu Fuguan formula can effectively reduce the 24-hour urine protein level of IMN patients, and the 24-hour urine protein level shows a significant downward trend with the extension of the treatment course; according to the IMN clinical efficacy standard, the total remission rate of patients receiving the Danzhu Fuguan formula treatment for 3 months is 37%, the total remission rate of patients receiving the treatment for 6 months is 50%, and the total remission rate of patients receiving the treatment for 12 months reaches 72%, and the remission rate increases significantly with the extension of the treatment course.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceuticals, specifically relating to the application of Danshu Fuyuan Formula in the preparation of drugs for treating idiopathic membranous nephropathy (IMN). Background Technology

[0002] Idiopathic membranous nephropathy, clinically known as primary nephrotic syndrome, is a type of nephrotic syndrome. It is generally caused by drug-induced irritation or may be due to pathogenic microbial infection. Patients may present with abnormal urination, such as hematuria, proteinuria, and edema. Some patients may also experience symptoms of venous thrombosis or infection. If these symptoms are not treated promptly, they can impair normal kidney function and, in severe cases, lead to kidney failure, posing a serious threat to health.

[0003] Traditional treatments may involve medications such as amoxicillin capsules and norfloxacin capsules, but these have certain side effects and are slow to take effect.

[0004] Therefore, based on this, the technical solution of the present invention is proposed. Summary of the Invention

[0005] To address the problems existing in the prior art, this invention provides an application of Danshu Fuyuan Formula in the preparation of drugs for treating idiopathic membranous nephropathy. The raw materials of Danshu Fuyuan Formula include: Danshen, Atractylodes macrocephala, Astragalus membranaceus, Lonicera japonica, Polygonum cuspidatum, Curcuma longa, Poria cocos, Coix lacryma-jobi, and Notopterygium incisum.

[0006] The formula uses Danshen (Salvia miltiorrhiza), which is bitter and slightly cold in nature, entering the pericardium and liver meridians. Its effects are similar to Siwu Decoction, cooling and promoting blood circulation, making it the principal herb. The assistant herbs are Atractylodes macrocephala (Bai Zhu) and Astragalus membranaceus (Huang Qi), which tonify the original qi; Lonicera japonica (Jin Yin Hua) and Polygonum cuspidatum (Hu Zhang), which clear heat from the blood vessels and disperse existing stagnation. The adjuvant herbs are Poria cocos (Fu Ling) and Coix lacryma-jobi (Yi Yi Ren), which strengthen the spleen and resolve phlegm, ensuring that Astragalus membranaceus and Atractylodes macrocephala do not become stagnant. Curcuma longa (Yu Jin), along with Lonicera japonica and Polygonum cuspidatum, assists Danshen in clearing heat, cooling the blood, and dispersing stagnation. Notopterygium incisum (Qiang Huo) serves as an adjuvant, its pungent and warm properties preventing the entire formula from becoming excessively cold, and its wind-dispelling properties promoting the upward movement of yang qi, while also restoring the spleen and stomach's qi circulation when combined with Astragalus membranaceus and Atractylodes macrocephala. The entire formula combines cold and hot herbs, tonifying and purging, with a scientifically sound and rational combination.

[0007] Preferably, the raw materials of the Danshu Fuyuan Formula also include one or more of the following: ginseng, fangji, coptis, angelica, dipsacus, epimedium, rehmannia, and bupleurum.

[0008] Preferably, the Danshu Fuyuan Formula comprises the following ingredients in parts by weight: 25-35 parts of Salvia miltiorrhiza, 10-15 parts of Atractylodes macrocephala, 25-35 parts of Astragalus membranaceus, 10-20 parts of Lonicera japonica, 10-20 parts of Polygonum cuspidatum, 10-20 parts of Curcuma longa, 10-15 parts of Poria cocos, 25-35 parts of Coix lacryma-jobi, and 10-15 parts of Notopterygium incisum.

[0009] Preferably, the Danshu Fuyuan Formula comprises the following ingredients in parts by weight: 30 parts of Salvia miltiorrhiza, 12 parts of Atractylodes macrocephala, 30 parts of Astragalus membranaceus, 15 parts of Lonicera japonica, 15 parts of Polygonum cuspidatum, 15 parts of Curcuma longa, 12 parts of Poria cocos, 30 parts of Coix lacryma-jobi, and 12 parts of Notopterygium incisum.

[0010] Preferably, the preparation method of the Danshu Fuyuan Formula is as follows:

[0011] The raw materials of Danshu Fuyuan Formula are pulverized and mixed to obtain a composition.

[0012] Alternatively, the raw materials of Danshu Fuyuan Formula can be mixed and then pulverized to obtain a composite material;

[0013] Alternatively, the raw materials of Danshu Fuyuan Formula can be extracted and refined using conventional processes to obtain one of the following: tablets, capsules, granules, pills, powders, or oral liquids.

[0014] Preferably, the drug is a drug that reduces urinary protein levels.

[0015] Preferably, the drug is a drug that increases plasma albumin and total protein levels.

[0016] Preferably, the drug is a drug for improving lipid metabolism disorders.

[0017] The beneficial effects of this invention are as follows:

[0018] Studies have shown that the Danshu Fuyuan Formula described in this invention can effectively reduce the 24-hour urinary protein level in IMN patients, and the 24-hour urinary protein level shows a significant decreasing trend with the extension of the treatment course. Furthermore, according to the clinical efficacy criteria for IMN, the overall remission rate of patients treated with Danshu Fuyuan Formula for 3 months was 37%, the overall remission rate for 6 months was 50%, and the overall remission rate for 12 months reached 72%, and the remission rate increased significantly with the extension of the treatment course.

[0019] The preferred embodiment of the present invention has the following advantages:

[0020] The modified Danshu Fuyuan formula, a traditional Chinese medicine treatment, is safe and effective in treating IMN. It can significantly reduce proteinuria in IMN patients and increase plasma albumin and total protein levels. At the same time, the modified Danshu Fuyuan formula can significantly improve lipid metabolism disorders in IMN patients.

[0021] The risk level and plasma albumin level of IMN patients are closely related to the efficacy of the modified Danshu Fuyuan formula. IMN patients with low risk and higher albumin levels have a better prognosis.

[0022] The Danshu Fuyuan Formula described in this invention provides a novel approach to the treatment of idiopathic membranous nephropathy. Detailed Implementation

[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0024] Example 1

[0025] This embodiment provides a method for preparing Danshu Fuyuan Formula, specifically: 25g of Danshen, 10g of Baizhu, 25g of Huangqi, 10g of Jinyinhua, 10g of Huzhang, 10g of Yujin, 10g of Fuling, 25g of Yiyiren and 10g of Qianghuo are pulverized and mixed to obtain a composition, which is then processed into tablets using conventional methods.

[0026] Example 2

[0027] This embodiment provides a method for preparing Danshu Fuyuan Formula, specifically: 35g of Danshen, 15g of Baizhu, 35g of Huangqi, 20g of Jinyinhua, 20g of Huzhang, 20g of Yujin, 15g of Fuling, 35g of Yiyiren and 15g of Qianghuo are mixed and pulverized to obtain a composition, which is then conventionally processed into capsules.

[0028] Example 3

[0029] This embodiment provides a method for preparing Danshu Fuyuan Formula, specifically: 30g of Danshen, 12g of Baizhu, 30g of Huangqi, 15g of Jinyinhua, 15g of Huzhang, 15g of Yujin, 12g of Fuling, 30g of Yiyiren and 12g of Qianghuo are mixed, and extracted and refined according to conventional processes to obtain granules.

[0030] Example 4

[0031] The difference between Example 4 and Example 3 is that the Danshu Fuyuan Formula is made into pills.

[0032] Example 5

[0033] The difference between Example 5 and Example 3 is that the Danshu Fuyuan Formula is made into a powder.

[0034] Example 6

[0035] The difference between Example 6 and Example 3 is that the Danshu Fuyuan Formula is made into an oral liquid.

[0036] Validation example (using the granules obtained in Example 3 as the validation object)

[0037] (a) Research Subjects

[0038] 1. Case source: IMN subjects and healthy individuals who received outpatient and inpatient treatment at Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Hospital of Chinese Medicine, and Fangshan District Hospital of Chinese Medicine in Beijing from December 2019 to December 2021.

[0039] 2. Diagnostic criteria:

[0040] 2.1 According to Wang Haiyan's 2008 edition of Nephrology (3rd edition), the onset is insidious, often without preceding infection, and the medical history and physical examination meet the diagnostic criteria for IMN.

[0041] 2.2. Referencing "Renal Biopsy Pathology (Third Edition)" edited by Zou Wanzhong in 2014. The renal tissue biopsy showed characteristics consistent with membranous nephropathy under light, electron, and immunofluorescence microscopy. Secondary membranous nephropathy (MN) was excluded, and the diagnosis was IMN, staged according to the Ehrenreich-Churg staging system.

[0042] Stage I: The glomerular basement membrane is basically normal, with small and scattered electron-dense deposits visible, irregularly distributed along the basement membrane epithelium; Stage II: The deposits increase compared to Stage I, with the formation of spikes and the disappearance of foot processes of epithelial cells; Stage III: The spikes fuse, and the deposited material is irregular and sparse, resembling worm-eaten material; Stage IV: The basement membrane thickens significantly and irregularly, and the density of the deposited electron-dense deposits decreases.

[0043] 2.3. Referring to the "Research on Macroscopic Diagnostic Criteria for Coronary Heart Disease with Phlegm-Blood Stasis Syndrome" published in the Chinese Journal of Integrated Traditional and Western Medicine in 2016, the TCM diagnostic criteria for IMN with Phlegm-Blood Stasis Syndrome were formulated, as shown in Table 1.

[0044] Table 1. Clinical diagnostic criteria for phlegm-blood stasis syndrome in idiopathic membranous nephropathy.

[0045]

[0046] (II) Research Plan

[0047] 1. Treatment plan:

[0048] 1.1 Basic Treatment: A low-salt, low-fat, and bland diet; use ACEIs and ARBs within the patient's tolerance range. Blood pressure should be controlled below 130 / 80 mmHg. If blood pressure has not reached the target level (target blood pressure: systolic blood pressure ≤130 mmHg and diastolic blood pressure ≤80 mmHg), CCB preparations, beta-blockers, and diuretics can be used in combination.

[0049] 1.2 Clinical Trial Methods: IMN Subject Group: Basic treatment + Danshu Fuyuan Formula (granules). Danshu Fuyuan Formula (granules) is composed of Danshen, Baizhu, Yujin, Yiyiren, Huangqi, Fuling, Huzhang, Jinyinhua, and Qianghuo. Efficacy: Tonifies Qi, resolves phlegm, invigorates blood, clears heat, and dissipates nodules. Dosage form: Granules. One dose of Chinese medicine is taken twice daily, morning and evening, for a course of 6 months.

[0050] 1.3. Concomitant Medication: For patients with hyperlipidemia, statins can be administered; for patients with hypoalbuminemia (plasma albumin below 25 g / L) and severe edema, human albumin can be administered intravenously; for patients with hypercoagulable states and thrombosis risk, medications such as PulseXueKang, aspirin, clopidogrel, or low molecular weight heparin can be given; for severe cases of respiratory tract infection, urinary tract infection, or gastrointestinal infection, antibiotics can be given. Mild respiratory tract infections can be treated with cold and fever-reducing granules, Jin Hua Qing Gan granules, etc.; mild gastrointestinal infections can be treated with berberine hydrochloride, Bacillus licheniformis, etc.; mild urinary tract infections can be treated with Re Lin Qing granules, Long Qing tablets, etc.; for patients with constipation, add Bian Tong capsules.

[0051] 2. Observation indicators and methods:

[0052] 2.1 Clinical efficacy observation indicators: (1) Primary efficacy indicators: 24-hour urine protein quantification (24hUTP), albumin (ALB). (2) Secondary efficacy indicators: serum total protein (TP), blood urea nitrogen (BUN), serum creatinine (Scr), estimated glomerular filtration rate (eGFR), eGFR was calculated using the CKD-EPI formula.

[0053] 2.2 TCM Syndrome Evaluation: The clinical symptoms of phlegm-blood stasis syndrome are quantified, including: abdominal distension, poor appetite, heaviness in the head and body, edema, obesity, dark lips or gums, dark or dull complexion, rough and dry skin, cyanosis of the extremities, sticky stools, drowsiness, and sticky mouth. These are categorized by severity as none, mild, moderate, and severe, scored as 0, 2, 4, and 6 points respectively. Tongue and pulse are not included in the score. Mild refers to occasional symptoms / mild sensations, moderate refers to persistent symptoms / significant sensations, and severe refers to symptoms affecting daily life. The TCM syndrome score is the sum of the scores for each symptom; a higher score indicates more severe symptoms. See Table 2 for details.

[0054] Table 2 Scoring Table for Phlegm-Blood Stasis Syndrome in Idiopathic Membranous Nephropathy

[0055]

[0056]

[0057] 3. Evaluation of therapeutic effect:

[0058] 3.1 Clinical efficacy evaluation criteria:

[0059] The IMN clinical efficacy criteria were established with reference to the 2012 KDIGO Clinical Practice Guidelines for Glomerulonephritis:

[0060] (1) Complete remission: urinary protein quantification is less than 0.3g / d, with an interval of at least 1 week, and the target is achieved twice; plasma albumin and serum creatinine are within the normal range.

[0061] (2) Partial relief: urinary protein quantification is less than 3.5g / d and has decreased by more than 50% from the peak value; the interval is at least 1 week, the target is achieved twice, plasma albumin is normal or improved, and serum creatinine is stable;

[0062] (3) Invalid: The above standards are not met.

[0063] 3.2 Traditional Chinese Medicine Syndrome Evaluation Standards:

[0064] Refer to the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine for the Treatment of Chronic Nephritis" in the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine (Trial)".

[0065] (1) Clinical cure: TCM clinical symptoms and signs disappear or basically disappear, and the symptom score decreases by ≥95%;

[0066] (2) Significant effect: Clinical symptoms and signs in TCM are significantly improved, and the symptom score is reduced by ≥70%;

[0067] (3) Effective: Clinical symptoms and signs in TCM patients improved, and the symptom score decreased by ≥30%;

[0068] (4) Ineffective: There is no significant improvement in the clinical symptoms and signs of TCM, or even aggravation, and the symptom score is reduced by less than 30%.

[0069] Note: The calculation formula (nimodipine method) is: [(pre-treatment score - post-treatment score) ÷ pre-treatment score] × 100%.

[0070] (III) Research Results

[0071] 1. Case Inclusion: This study included 10 healthy subjects and 65 subjects from the IMN (Integrated Medical Network) of Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Hospital of Chinese Medicine, and Fangshan District Hospital of Chinese Medicine, Beijing. Six subjects were mistakenly admitted, and five were lost to follow-up after three months of treatment. Ultimately, 59 subjects were included in the FAS (Functional Analysis Set) for statistical analysis, 54 subjects were included in the PPS (Programmable Statistical Analysis Set), and 59 subjects were included in the SS (Safety Analysis Set).

[0072] 2. General and clinical information:

[0073] 2.1 Gender and Age: Among the 54 IMN subjects, 29 were male (53.7%) and 25 were female (46.3%), with a male-to-female ratio of 1.16:1. The healthy control group consisted of 10 subjects, 5 males (50%) and 5 females (50%), with a male-to-female ratio of 1:1. Chi-square test showed no statistically significant difference in gender between the two groups (P > 0.05). The IMN group ranged in age from 20 to 77 years, with a mean age of 53.07 ± 12.86 years. The healthy control group ranged in age from 28 to 63 years, with a mean age of 46.6 ± 10.53 years. t-test showed no statistically significant difference in age between the two groups (P > 0.05). Specific results are shown in Table 3.

[0074] Table 3 Comparison of basic data between the IMN group and the healthy control group (x±s)

[0075]

[0076] Note: When comparing the IMN group with the healthy group, P < 0.05 is considered statistically significant.

[0077] 2.2 CKD staging: Among the 54 IMN subjects, 36 subjects were CKD stage 1, accounting for 66.7%; 14 subjects were CKD stage 2, accounting for 25.9%; and 4 subjects were CKD stage 3, accounting for 7.4%.

[0078] 2.3 Risk assessment: Of the 54 IMN subjects, 26 were at low risk, accounting for 48.1%; 23 were at medium risk, accounting for 42.6%; and 5 were at high risk, accounting for 9.3%.

[0079] 2.4 Pathological staging: Among the 54 IMN subjects, there were 19 subjects in stage I, 13 subjects in stage I-II, 11 subjects in stage II, 2 subjects in stage II-III, and 9 subjects with atypical membrane kidneys.

[0080] 2.5 Among the 54 IMN subjects, 5 cases, accounting for 9.3% of the total, presented with proteinuria within the range of nephrotic syndrome.

[0081] 2.6 Distribution of comorbidities: Among the 54 IMN subjects, 16 had hypertension, accounting for 29.6%; 8 had hyperlipidemia, accounting for 14.8%; 7 had diabetes, accounting for 13.0%; 3 had carotid and lower extremity arteriosclerosis, accounting for 5.6%; 2 had inflammatory skin diseases such as psoriasis and eczema, accounting for 3.7%; and 1 each had coronary heart disease, lacunar infarction, respiratory diseases, hypokalemia, benign prostatic hyperplasia, and femoral head necrosis, accounting for 1.9%.

[0082] 3. FAS analysis set:

[0083] 3.1 Evaluation of the main efficacy indicators of Danshu Fuyuan Formula (granules) in the treatment of IMN: The study compared the changes in 24hUTP and ALB levels before and after treatment in IMN subjects. The results showed that after 3 and 6 months of treatment with Danshu Fuyuan granules, the 24hUTP level of IMN subjects was significantly lower than that before treatment, and the difference was statistically significant (P < 0.01). After 6 months of treatment, the ALB level was slightly higher than that before treatment, but the difference was not statistically significant (P > 0.05). The specific results are shown in Table 4.

[0084] Table 4. 24h UTP and ALB levels before and after treatment in 59 IMN subjects (x±s)

[0085]

[0086] Note: Compared with the IMN group before treatment, *P<0.05, **P<0.01, the same below.

[0087] 3.2 Clinical remission rate of Danshu Fuyuan Formula (granules) in the treatment of IMN: After 3 months of treatment with Danshu Fuyuan granules, 3 out of 59 IMN subjects achieved complete remission (response rate 5.1%), 17 achieved partial remission (response rate 28.8%), and the overall remission rate was 33.9%. After 6 months of treatment, 5 out of 5 subjects achieved complete remission (response rate 8.5%), 23 achieved partial remission (response rate 39.0%), and the overall remission rate was 47.5%.

[0088] 4. PPS Analysis Set:

[0089] 4.1 Evaluation of the main efficacy indicators of Danshu Fuyuan Formula (granules) in the treatment of IMN: The study compared the changes in 24hUTP and ALB levels before and after treatment in IMN subjects. Results showed that after 3 months of treatment with Danshu Fuyuan granules, the 24hUTP level in IMN subjects decreased compared to before treatment, with a statistically significant difference (P < 0.05). The efficacy after 6 months of treatment was better than that after 3 months, with a more significant decrease in 24hUTP, and the difference was statistically significant (P < 0.01). The ALB level in 54 subjects increased slightly compared to before treatment, but the difference was not statistically significant (P > 0.05). Specific results are shown in Table 5.

[0090] Table 5. 24h UTP and ALB levels before and after treatment in 54 IMN subjects (x±s)

[0091]

[0092] 4.2 Evaluation of secondary efficacy endpoints of Danshu Fuyuan Formula (granules) in the treatment of IMN: The study compared the changes in TP, BUN, CREA, and eGFR levels before and after treatment in IMN subjects. The results showed that after 3 and 6 months of treatment with Danshu Fuyuan granules, there were no statistically significant differences in TP, BUN, CREA, and eGFR levels compared with those before treatment (P>0.05). Specific results are shown in Table 6.

[0093] Table 6. TP, BUN, CREA, and eGFR levels before and after treatment in 54 IMN subjects (x±s)

[0094]

[0095] 4.3 Clinical remission rate of Danshu Fuyuan Formula (granules) in the treatment of IMN: After 3 months of treatment with Danshu Fuyuan granules, 3 IMN subjects achieved complete remission (response rate 5.6%), 17 achieved partial remission (response rate 31.4%), and the overall remission rate was 37.0%. After 6 months of treatment, 5 subjects achieved complete remission (response rate 9.3%), 24 achieved partial remission (response rate 40.7%), and the overall remission rate was 50.0%.

[0096] 4.4 Evaluation of TCM Syndrome Scores and Efficacy of Danshu Fuyuan Granules in the Treatment of IMN: This study compared the changes in TCM syndrome scores before and after treatment in IMN subjects. Results showed that the TCM syndrome scores of IMN subjects significantly decreased after 3 and 6 months of treatment with Danshu Fuyuan Granules (P < 0.01). The decrease in TCM syndrome scores after 6 months of treatment was more significant than that after 3 months of treatment (P < 0.01). After 3 months of treatment with Danshu Fuyuan Granules, 2 IMN subjects achieved clinical cure, 0 showed significant improvement, 15 showed improvement, and 37 showed no improvement, with a total effective rate of 31.5%. After 6 months of treatment, 6 subjects achieved clinical cure, 9 showed significant improvement, 29 showed improvement, and 10 showed no improvement, with a total effective rate of 81.5%. Specific results are shown in Table 7.

[0097] Table 7. Results of TCM syndrome scores (x±s) before and after treatment in 54 IMN subjects.

[0098]

[0099] Note: Compared with the IMN group before treatment, *P<0.05, **P<0.01; compared with 3 months of treatment at 6 months, △ P < 0.05 △△ P < 0.01.

[0100] 4.5 One-year follow-up: Of the 54 IMN subjects, 25 had 12-month follow-up data. The 24-hour UTP results of these 25 subjects were analyzed, and the specific results after one year are shown in Table 8.

[0101] Table 8. 24hUTP results (x±s) of 25 subjects

[0102]

[0103] After 3 months of treatment with Danshu Fuyuan Granules, 1 of the 25 subjects achieved complete remission (4% remission rate), 10 achieved partial remission (40% remission rate), and the overall remission rate was 44.0%. After 6 months of treatment, 3 subjects achieved complete remission (12% remission rate), 8 achieved partial remission (32% remission rate), and the overall remission rate was 44.0%. A one-year follow-up revealed that among the 25 subjects, 6 achieved complete remission (24% remission rate), 12 achieved partial remission (48% remission rate), and the overall remission rate was 72%.

[0104] 4.6 Evaluation of the efficacy of Danshu Fuyuan Granules in treating IMN (Intra-Infectious Disease) grouping:

[0105] 4.6.1 Baseline Data Comparison: Patients were grouped according to whether they had previously used hormones or immunosuppressants. The group with a history of hormone or immunosuppressant use consisted of patients who had received standardized treatment in the nephrology department of a tertiary hospital for more than 6 months without achieving clinical remission. Baseline levels were compared between the two groups.

[0106] The subjects were grouped according to their prior use of hormones or immunosuppressants; 24 had previously used hormones or immunosuppressants, and 30 had not. No statistically significant differences were found between the two groups in terms of gender, age, CKD stage, risk assessment, pathological stage composition ratio, and laboratory test results (24hUTP, ALB, TP, BUN, CREA, eGFR) (P > 0.05). Therefore, the group comparisons were comparable. Specific results are shown in Table 9.

[0107] Table 9. Comparison of baseline data of the two groups of IMN subjects

[0108]

[0109]

[0110] 4.6.2 Previous use of hormones or immunosuppressants: Among the 24 subjects with a history of using hormones or immunosuppressants, 21 had used hormone drugs, accounting for 87.5%; 8 had used cyclophosphamide, accounting for 33.3%; 8 had used cyclosporine, accounting for 33.3%; 6 had used triptolide preparations, accounting for 25%; 4 had used tacrolimus, accounting for 16.7%; 2 had used leflunomide, accounting for 8.3%; 2 had used rituximab, accounting for 8.3%; and 1 had used mycophenolate mofetil, accounting for 4.2%.

[0111] 4.6.3 Comparison of clinical remission rates between the two groups: After 3 months of treatment with Danshu Fuyuan granules, the clinical remission rate was 46.7% in subjects who did not use hormones or immunosuppressants, and 25% in subjects who had previously used hormones or immunosuppressants. There was no statistically significant difference in clinical remission rates between the two groups (P>0.05).

[0112] After 6 months of treatment with Danshu Fuyuan granules, the clinical remission rate was 63.3% in subjects who did not use hormones or immunosuppressants, and 33.3% in subjects who had previously used hormones or immunosuppressants. The difference in clinical remission rates between the two groups was statistically significant (P < 0.05). With prolonged treatment, subjects who did not use hormones or immunosuppressants showed better efficacy, as detailed in Table 10.

[0113] Table 10 Clinical response rates in the two groups

[0114]

[0115] Note: *P<0.05 compared with those who have previously used hormones or immunosuppressants.

[0116] 5. Safety evaluation:

[0117] 5.1 Effect of Danshu Fuyuan Granules on Safety Indicators: ALT and AST levels of IMN subjects were compared after 3 months and 6 months of treatment. The results showed that there was no statistically significant difference in ALT and AST levels before and after treatment (P>0.05). The specific results are shown in Table 11.

[0118] Table 11 ALT and AST levels in IMN subjects before and after treatment (x±s)

[0119]

[0120] 5.2 Complications during treatment: During the follow-up period, 4 cases developed abnormal liver function and 1 case developed a respiratory infection. No serious adverse events occurred.

[0121] (iv) Modified Danshu Fuyuan Formula for the Intervention of Idiopathic Membranous Nephropathy

[0122] 1. Treatment plan:

[0123] In addition to basic treatment, all subjects received Danshu Fuyuan Formula, which was modified according to the emphasis of the pathogenesis.

[0124] Composition and preparation method of Danshu Fuyuan Formula: Astragalus membranaceus, Salvia miltiorrhiza, Atractylodes macrocephala, Curcuma longa, Coix lacryma-jobi, Poria cocos, Polygonum cuspidatum, Lonicera japonica, Notopterygium incisum. Decoct in water and take twice daily, after breakfast and dinner. Preparation method: The decoction room of Xiyuan Hospital will prepare the medicine. Each dose will yield 400 ml of decoction, divided into two packets. Dosage: 200 ml twice daily, taken warm half an hour after breakfast and dinner.

[0125] Treatment plan with modifications: For those with obvious fatigue, shortness of breath, and qi deficiency, add ginseng or codonopsis, dodder seed, euryale seed, and prepared licorice root; for those with obvious blood stasis, add angelica, red peony root, and chuanxiong rhizome; for those with obvious phlegm-heat, add coptis, scutellaria, and forsythia, and remove notopterygium root; for those with chest and rib distension and qi stagnation, add corydalis rhizome, bupleurum root, and immature bitter orange peel; for those with combined lower back pain, weak legs, and kidney deficiency, add dipsacus root and eucommia bark; for those with yin deficiency such as dry mouth, dry throat, and dry tongue, add rehmannia root, cornus fruit, ophiopogon root, and scrophularia root, and remove notopterygium root; for those with yang deficiency such as aversion to cold, cold limbs, and cold extremities, add epimedium, curculigo, prepared aconite root, and ginger; for those with obvious edema, add tangerine peel, mulberry bark, and ginger peel.

[0126] Basic treatment: A low-salt, low-fat, and bland diet; use ACEIs and ARBs at a tolerable dose. Blood pressure should be controlled below 130 / 80 mmHg. If blood pressure has not reached the target level (target blood pressure: systolic blood pressure ≤130 mmHg and diastolic blood pressure ≤80 mmHg), CCB preparations or beta-blockers may be used in combination.

[0127] Concomitant medication guidelines: For patients with hyperlipidemia, statins may be given; for patients with hypercoagulable states and thrombosis risk, PulseXueKang may be given; for patients with respiratory tract infections, urinary tract infections, or gastrointestinal infections, antibiotics may be given.

[0128] Follow-up period: 6 consecutive months of observation. Visit time: before treatment and at 3 and 6 months after treatment.

[0129] 2. Efficacy evaluation indicators:

[0130] Main therapeutic indicators: 24-hour urine protein quantification, serum albumin.

[0131] Secondary efficacy endpoints: serum total protein, eGFR, serum creatinine, total cholesterol, and triglycerides.

[0132] 3. Results Study:

[0133] 3.1 Case Inclusion: A total of 303 patients with IMN were included in the clinical study from Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Hospital of Chinese Medicine, and Fangshan District Hospital of Chinese Medicine, Beijing. 20 cases were excluded (17 incorrectly included, 3 misdiagnosed). Among these, 32 patients had a follow-up period of less than 6 months, and their last primary efficacy endpoint results were carried over to the FAS set for statistical analysis (283 cases). 251 patients completed the 6-month follow-up and were included in the PPS set for statistical analysis. 215 patients who received at least one treatment and had recorded post-treatment safety indicators were included in the SS set for safety analysis.

[0134] 3.2 General and Clinical Information:

[0135] 3.2.1 Gender and Age: Of the 251 patients, 136 were male and 115 were female. The patients' ages ranged from 18 to 70 years old, with 14 patients aged 18-30 (5.6%), 84 patients aged 31-50 (33.5%), and 153 patients aged 51-70 (61.0%).

[0136] 3.2.2 CKD staging: CKD staging of 251 IMN patients: 165 cases (65.7%) were CKD stage 1; 67 cases (26.7%) were CKD stage 2; and 19 cases (7.6%) were CKD stage 3.

[0137] 3.2.3 Risk assessment: Of the 251 patients with IMN, 141 (56.2%) were at low risk; 73 (29.1%) were at medium risk; and 37 (14.7%) were at high risk.

[0138] 3.2.4 Pathological staging: Among the 251 IMN patients, the highest proportion was stage I with 86 cases (34.3%); followed by stage II patients with a total of 69 cases (27.5%); stage I-II with 43 cases (17.1%); stage II-III with 11 cases (4.4%); and atypical membranous nephropathy with 42 cases (16.7%).

[0139] 3.2.5 Distribution of nephrotic syndrome: Among the 251 patients with IMN, 89 patients presented with proteinuria within the scope of nephrotic syndrome, accounting for 35.5% of the total.

[0140] 3.2.6 Distribution of comorbidities: Among the patients included in the study, 125 had hypertension, accounting for 49.8%; 28 had diabetes, accounting for 11.2%; 10 had coronary heart disease, accounting for 4.0%; 13 had hyperuricemia, accounting for 5.2%; 9 had cerebrovascular disease, accounting for 3.6%; 6 had rheumatoid arthritis, accounting for 2.4%; 6 had psoriasis, accounting for 2.4%; and 5 had autoimmune thyroid disease, accounting for 2.0%.

[0141] 3.2.7. Modifications to the Danshu Fuyuan Formula: All subjects received the Danshu Fuyuan Formula as part of their basic treatment, with modifications made according to the specific pathogenesis. The most frequent addition was ginseng or codonopsis, accounting for 99.2%; followed by fangji (Stephania tetrandra), accounting for 93.2%. Details are shown in Table 12.

[0142] Table 12 Modifications to the Danshu Fuyuan Formula

[0143]

[0144] 3.3 FAS Analysis Set:

[0145] 3.3.1 Evaluation of the main efficacy indicators of Danshu Fuyuan Formula for treating IMN: All 283 IMN patients received Danshu Fuyuan Formula with added ingredients. After 3 and 6 months of treatment, the 24hUTP of IMN patients decreased compared with that before treatment, and the difference was statistically significant (P < 0.01). ALB showed a significant upward trend compared with that before treatment, and the mean value was close to the normal level, and the difference was statistically significant (P < 0.01), as shown in Tables 13 and 14.

[0146] Table 13. Statistical data of 283 IMN patients before and after 24hUTP treatment [M(Q1~Q3)]

[0147]

[0148] Table 14. Statistical data of 283 IMN patients before and after 24hUTP treatment (x±s)

[0149]

[0150] 3.3.2 Clinical remission rate of modified Danshu Fuyuan formula for treating IMN: The effective rates of 283 patients treated for 3 months and 6 months were 34.6% and 56.9%, respectively, as shown in Table 15.

[0151] Table 15 Clinical response rate in 283 patients with IMN [cases (%)]

[0152]

[0153] 3.4 PPS Analysis Set:

[0154] 3.4.1 Evaluation of the main efficacy indicators of Danshu Fuyuan Formula for treating IMN: All 251 IMN patients received Danshu Fuyuan Formula with added ingredients. After 3 and 6 months of treatment, the 24hUTP of IMN patients decreased compared with that before treatment, and the difference was statistically significant (P < 0.01). ALB showed a significant upward trend compared with that before treatment, and the mean value was close to the normal level, and the difference was statistically significant (P < 0.01), as shown in Tables 16 and 17.

[0155] Table 16. Statistical data of 251 IMN patients before and after 24hUTP treatment [M(Q1~Q3)]

[0156]

[0157] Table 17. Statistical data of 251 IMN patients before and after ALB treatment (x±s)

[0158]

[0159] 3.4.2 Evaluation of secondary efficacy indicators of modified Danshu Fuyuan Formula for the treatment of IMN: After 3 months and 6 months of treatment, the total phosphorus (TP) of IMN patients increased compared with that before treatment, and the difference was statistically significant (P<0.01).

[0160] After 3 and 6 months of treatment, patients' Scr decreased compared with before treatment, and eGFR increased compared with the baseline results before treatment. The differences were statistically significant (P < 0.01), as shown in Tables 18 and 19.

[0161] Table 18. TP, TC, and eGFR levels before and after treatment in 251 IMN patients (x±s)

[0162]

[0163] Table 19. TG and Scr levels before and after treatment in 251 IMN patients [M(Q1~Q3)]

[0164]

[0165] 3.4.3 Clinical remission rate of IMN with modified Danshu Fuyuan Formula as the main treatment: In this study, the effective rates of 251 patients after 3 months and 6 months of treatment were 37.5% and 61.4%, respectively, as shown in Table 20.

[0166] Table 20 Clinical remission rate of 251 patients with IMN treated with modified Danshu Fuyuan Formula

[0167]

[0168] 3.5 Safety evaluation of modified Danshu Fuyuan formula for treating IMN:

[0169] 3.5.1 Safety indicators: No significant abnormalities were observed in liver function before treatment and at 3 and 6 months of treatment, and the differences were not statistically significant (P>0.05), as shown in Table 21.

[0170] Table 21. AST and ALT levels in IMN patients before and after treatment (x±s)

[0171]

[0172] 3.5.2 Complications during treatment: During the follow-up period, 5 cases of infection occurred, including 2 cases of pulmonary infection, 2 cases of gastroenteritis, and 1 case of urinary tract infection; 1 case of heart failure; 1 case of atrial fibrillation; 5 cases of abnormal liver function; and 1 case of rash.

[0173] 3.6 Analysis of factors influencing the clinical efficacy of modified Danshu Fuyuan formula:

[0174] 3.6.1 Univariate Analysis: Patients were grouped according to whether they achieved remission. The following factors were compared between the two groups: gender, age, pathological type, risk level, CKD stage, history of nephrotic syndrome, pre-treatment serum albumin, pre-treatment serum total protein, pre-treatment 24-hour urinary protein, pre-treatment cholesterol, pre-treatment triglycerides, pre-treatment serum creatinine, pre-treatment glomerular filtration rate, history of immunosuppressive therapy, and medical history. Factors with statistically significant differences were screened. It can be seen that, after univariate screening, CKD stage, clinical manifestation of nephrotic syndrome, risk stratification, pre-treatment 24-hour UTP, pre-treatment ALB, pre-treatment TP, pre-treatment SCR, and pre-treatment eGFR may be influencing factors on the clinical efficacy of Jiawei Danshu Fuyuan Fang and need to be retained for subsequent multivariate analysis. See Tables 22-24 for details.

[0175] Table 22. Univariate analysis results of factors influencing clinical efficacy in 251 patients with IMN.

[0176]

[0177]

[0178] Table 23. Univariate analysis results of factors influencing clinical efficacy in 251 patients with IMN [M(Q1~Q3)]

[0179]

[0180] Table 24. Univariate analysis results of factors influencing clinical efficacy in 251 patients with IMN (x±s)

[0181]

[0182] 3.6.2 Multivariate analysis: Eight variables were used as independent variables, including CKD stage, clinical manifestation of nephrotic syndrome, risk level, 24h-UTP before treatment, ALB before treatment, TP before treatment, SCR before treatment, and eGFR before treatment, and whether the condition was in remission was used as the dependent variable. A binary logistic regression analysis was performed, as shown in Table 25.

[0183] Table 25. Multivariate analysis results of factors influencing clinical efficacy in 251 IMN patients.

[0184]

[0185] The Hosmer-Lemmeshow goodness-of-fit test (P=0.263>0.05) indicates that the regression model fits the actual data well, as shown in Table 26.

[0186] Table 26 Hosmer-Lemeshow Test

[0187]

[0188] The calculation results in the table above clearly show that:

[0189] CKD stage and clinical presentation of nephrotic syndrome were not independent prognostic factors for IMN (P>0.05);

[0190] Pre-treatment 24hUTP, pre-treatment TP, pre-treatment SCR, and pre-treatment eGFR were not independent prognostic factors for IMN (P>0.05);

[0191] IMN risk level was an independent prognostic factor for IMN (P=0.029<0.05); pre-treatment ALB and the efficacy of modified Danshu Fuyuan Formula for treating IMN were significantly positively correlated, suggesting that pre-treatment ALB was an independent prognostic factor for IMN (P<0.01).

[0192] (V) Conclusion

[0193] 1. Evaluation of the efficacy of Danshu Fuyuan Formula in Western medicine:

[0194] Danshu Fuyuan Formula can effectively reduce the 24-hour urinary protein level in IMN patients, and the 24-hour urinary protein level shows a significant downward trend with the extension of the treatment course; however, it has no significant intervention effect on the plasma albumin and total protein levels in IMN patients.

[0195] According to the IMN clinical efficacy criteria, the overall response rate was 37% for patients treated with Danshu Fuyuan Fang for 3 months, 50% for those treated for 6 months, and 72% for those treated for 12 months. The study showed that the response rate increased significantly with the extension of treatment duration.

[0196] Based on whether or not they had previously used hormones or immunosuppressants, after 3 months of treatment with Danshu Fuyuan Fang, the clinical remission rate was 46.7% for subjects who had not used hormones or immunosuppressants, and 25% for subjects who had previously used hormones or immunosuppressants. After 6 months of treatment, the clinical remission rate was 63.3% for subjects who had not used hormones or immunosuppressants, and 33.3% for subjects who had previously used hormones or immunosuppressants. The study indicates that with prolonged treatment, subjects who had not used hormones or immunosuppressants showed better efficacy.

[0197] 2. Evaluation of Traditional Chinese Medicine Syndrome Score for Danshu Fuyuan Formula:

[0198] Studies have shown that Danshu Fuyuan Formula can effectively improve the clinical symptoms of phlegm and blood stasis syndrome in traditional Chinese medicine, such as abdominal distension, poor appetite, heaviness in the head and body, edema, obesity, dark lips or gums, dark or dull complexion, rough and dry skin, cyanosis of the extremities, sticky stools, drowsiness, and sticky mouth. The longer the course of treatment, the more obvious the improvement in symptoms.

[0199] 3. Safety evaluation of Danshu Fuyuan Formula:

[0200] Studies have shown that Danshu Fuyuan Formula is safe and effective in treating IMN syndrome caused by phlegm and blood stasis, with no obvious adverse reactions and no effect on liver function.

[0201] 4. Modified Danshu Fuyuan Formula for the Intervention of Idiopathic Membranous Nephropathy:

[0202] 4.1 The modified Danshu Fuyuan Formula is a safe and effective treatment for IMN, which can significantly reduce proteinuria and increase plasma albumin and total protein levels in IMN patients. At the same time, the modified Danshu Fuyuan Formula can significantly improve lipid metabolism disorders in IMN patients.

[0203] 4.2 The risk level and plasma albumin level of IMN patients are closely related to the efficacy of modified Danshu Fuyuan Formula. IMN patients with low risk and higher albumin levels have a better prognosis.

[0204] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. The application of a Danshu Fuyuan formula in the preparation of drugs for treating idiopathic membranous nephropathy, characterized in that, The Danshu Fuyuan Formula is composed of the following ingredients in parts by weight: 25-35 parts of Danshen, 10-15 parts of Baizhu, 25-35 parts of Huangqi, 10-20 parts of Jinyinhua, 10-20 parts of Huzhang, 10-20 parts of Yujin, 10-15 parts of Fuling, 25-35 parts of Yiyiren, and 10-15 parts of Qianghuo.

2. The application according to claim 1, characterized in that, The Danshu Fuyuan Formula is composed of the following ingredients in parts by weight: 30 parts of Danshen, 12 parts of Baizhu, 30 parts of Huangqi, 15 parts of Jinyinhua, 15 parts of Huzhang, 15 parts of Yujin, 12 parts of Fuling, 30 parts of Yiyiren, and 12 parts of Qianghuo.

3. The application according to claim 1 or 2, characterized in that, The preparation method of the Danshu Fuyuan Formula is as follows: The raw materials of Danshu Fuyuan Formula are pulverized and mixed to obtain a composition. Alternatively, the raw materials of Danshu Fuyuan Formula can be mixed and then pulverized to obtain a composite material; Alternatively, the raw materials of Danshu Fuyuan Formula can be extracted and refined using conventional processes to obtain one of the following: tablets, capsules, granules, pills, powders, or oral liquids.

4. The application according to claim 1 or 2, characterized in that, The drug in question is one that reduces proteinuria levels.

5. The application according to claim 1 or 2, characterized in that, The drug is used to increase plasma albumin and total protein levels.

6. The application according to claim 1 or 2, characterized in that, The drug in question is for improving lipid metabolism disorders.

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