A traditional chinese medicine composition for treating immune-related glomerular diseases and its application

Abstract

The traditional Chinese medicine composition disclosed by the invention adopts a method of warming yang and relieving exterior syndrome to achieve the effects of freeing lung and inducing diuresis, tonifying spleen and warming kidney, and strengthening defensive qi and diminishing swelling, and is prepared to be used for treating the immune-related glomerular diseases, especially membranous nephropathy and minimal lesion nephropathy. The traditional Chinese medicine composition is a pure traditional Chinese medicine preparation, clinical observation and in-vivo and in-vitro basic tests show that the prescription can remarkably improve edema, massive proteinuria, hypoproteinemia and the like, meanwhile, compared with the prior art, the traditional Chinese medicine composition has a better effect on symptoms such as stomachache and diarrhea, has an anti-inflammatory effect and does not have obvious side effects in clinical application. The traditional Chinese medicine composition has an excellent effect on patients suffering from membranous nephropathy and minimal lesion nephropathy caused by syndrome differentiation of spleen-kidney yang deficiency, internal rheumatism and water-fluid overflowing.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202511901692.1 (22) Application Date 2025.12.16 (71) Applicant: Beijing Hospital of Traditional Chinese Medicine, Capital Medical University Address: No. 23, Meishuguan Houjie, Dongcheng District, Beijing 100010, China (72) Inventor: Liu Baoli (74) Patent Agency: Beijing Xianghe Intellectual Property Agency (General Partnership) 11893 Patent Attorney: Feng Mingyan (51) Int.Cl. A61K 36 / 9068 (2006.01) A61K 36 / 17 (2006.01) A61K 36 / 481 (2006.01) A61P 13 / 12 (2006.01) (54) Invention Title: A Traditional Chinese Medicine Composition for Treating Immune-Related Glomerular Diseases and Its Application (57) Abstract: This invention employs the method of warming yang and relieving exterior syndromes to achieve the effects of clearing the lungs and promoting diuresis, strengthening the spleen and warming the kidneys, and strengthening the defensive qi and reducing swelling. A traditional Chinese medicine composition is proposed for treating immune-related glomerular diseases, particularly membranous nephropathy and minimal change disease. The drug composition in this invention is a pure traditional Chinese medicine preparation. Clinical observation and in vitro and in vivo basic tests show that the prescription can significantly improve edema, massive proteinuria, and hypoalbuminemia. It also shows better efficacy than existing technologies in treating symptoms such as abdominal pain and diarrhea, and has anti-inflammatory effects. No obvious side effects were found in clinical application. It is particularly effective for patients with membranous nephropathy and minimal change disease whose TCM diagnosis is spleen and kidney yang deficiency, internal wind-dampness, and water retention. Claims (2 pages), Description (14 pages), Drawings (3 pages), CN 121401392 A 2026.01.27 CN 1 21 40 13 92 A 1. A traditional Chinese medicine composition for treating kidney disease, the traditional Chinese medicine composition comprising the following traditional Chinese medicines in parts by weight: 3-30 parts Ephedra, 5-120 parts Astragalus, 3-30 parts Aconitum carmichaelii (processed), 5-60 parts Poria cocos, 5-50 parts Cinnamomum cassia, 5-60 parts Atractylodes macrocephala (fried), 5-60 parts Stephania tetrandra (processed), 3-50 parts Glycyrrhiza uralensis (processed), 3-50 parts Zingiber officinale (fresh), and 3-50 parts Ziziphus jujuba. The traditional Chinese medicine composition can significantly improve edema, massive proteinuria, and hypoalbuminemia, and also has significant therapeutic effects on symptoms such as abdominal pain and diarrhea, and has anti-inflammatory effects. 2. The traditional Chinese medicine composition as described in claim 1, wherein the traditional Chinese medicine composition comprises the following traditional Chinese medicines in parts by weight: 5-25 parts ephedra, 15-100 parts astragalus, 5-25 parts prepared aconite root, 10-50 parts poria cocos, 10-30 parts cinnamon twig, 10-30 parts stir-fried atractylodes macrocephala, 8-30 parts bupleurum chinense root, 8-30 parts prepared licorice root, 5-40 parts fresh ginger, and 5-40 parts jujube. 3. The traditional Chinese medicine composition as described in claim 1, wherein the traditional Chinese medicine composition comprises the following traditional Chinese medicines in parts by weight: 6-15 parts ephedra...The ingredients are: 20-60 parts Astragalus membranaceus, 8-20 parts Aconitum carmichaelii (processed), 15-30 parts Poria cocos, 10-20 parts Cinnamomum cassia, 15-30 parts Atractylodes macrocephala (fried), 9-25 parts Stephania tetrandra (processed), 6-15 parts Glycyrrhiza uralensis (processed), 9-15 parts Zingiber officinale (fresh), and 9-15 parts Ziziphus jujuba. 4. The traditional Chinese medicine composition as described in claim 1, wherein 1-2 medicinal and edible herbs may be added according to specific symptoms. For example, for poor sleep, Albizia julibrissin bark or Ziziphus jujuba seed may be added as adjuvant herbs; for constipation, hemp seed or rhubarb (processed with wine) may be added; for blood stasis, Panax notoginseng powder, Curcuma zedoaria, or Hirudo medicinalis may be added; for significant sweating, ginseng or Paeonia lactiflora may be added. 5. A traditional Chinese medicine preparation for treating kidney disease, said preparation comprising the following ingredients: 3-30 parts ephedra, 3-30 parts prepared aconite root, 5-120 parts astragalus, 5-60 parts poria cocos, 5-50 parts cinnamon twig, 5-60 parts bupleurum root, 5-60 parts stir-fried atractylodes macrocephala, 3-50 parts ginger, 3-50 parts jujube, 3-50 parts prepared licorice root, and excipients required for preparing the corresponding traditional Chinese medicine dosage form. 6. The traditional Chinese medicine preparation according to claim 5, wherein the dosage form is one of the following: decoction, tablets, pills, capsules, granules, powdered tablets, oral liquid, or compound traditional Chinese medicine. 7. A method for preparing a traditional Chinese medicine composition for treating kidney disease, wherein the traditional Chinese medicine composition is obtained by the following preparation method: Preparation method 1 includes the following steps: S01, take the prescribed amount of black aconite slices, soak in water, adjust the pH, decoct and extract, ultrafilter through an ultrafiltration membrane to obtain the extract, concentrate and dry to obtain powder A; S02, take the prescribed amount of ephedra and fangji, pulverize, soak in water, adjust the pH, extract by ultrasonication to obtain the extract, concentrate and dry to obtain powder B; S03, take the prescribed amount of ginger, wash and juice, filter to obtain ginger juice, set aside; S04, take the prescribed amount of astragalus, poria, cinnamon twig, stir-fried atractylodes macrocephala, prepared licorice root and jujube, decoct in water, filter, concentrate to a clear extract, dry to obtain powder C; S05, mix powders A, B and C obtained in steps S01, S02 and S04, add ginger juice obtained in step S03, mix well, dry, pulverize to obtain the traditional Chinese medicine composition; Preparation method 2 includes the following steps: S01, Take the prescribed amounts of Aconitum carmichaelii slices, Ephedra sinica, and Stephania tetrandra, pulverize and mix them, soak them in water, adjust the pH, extract with ultrasound to obtain an extract, concentrate and dry to obtain powder A; S02, Take the prescribed amount of fresh ginger, wash and juice it, filter to obtain ginger juice, and set aside; S03, Take the prescribed amounts of Astragalus membranaceus, Poria cocos, Cinnamomum cassia, stir-fried Atractylodes macrocephala, prepared Glycyrrhiza uralensis, and jujube, decoct them in water, filter and concentrate to a clear paste, dry to obtain powder B; S04, Mix powder A and powder B from steps S01 and S03 evenly, add the ginger juice obtained in step S02, mix well, dry, pulverize, and obtain the traditional Chinese medicine composition. 8. The use of the traditional Chinese medicine composition of claim 1 or the traditional Chinese medicine preparation of claim 5 in the preparation of a medicament for treating immune nephropathy. Claims 1 / 2 pages 2 CN 121401392 A9. The application as described in claim 8, wherein the immune nephropathy includes primary immune nephropathy and secondary immune nephropathy. 10. The application as described in claim 8, wherein the primary immune nephropathy includes membranous nephropathy and minimal change disease, wherein the TCM syndrome differentiation of membranous nephropathy is spleen and kidney yang deficiency with water retention, and the TCM syndrome differentiation of minimal change disease is spleen and kidney yang deficiency with water retention. Claims 2 / 2 pages 3 CN 121401392 A Traditional Chinese Medicine Composition for Treating Immune-Related Glomerular Diseases and Its Application Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to a traditional Chinese medicine composition and its application. Background Art

[0002] The kidney, as an important metabolic and filtering organ in the human body, relies on the precise regulation of the immune system for its functional homeostasis. Autoimmune nephropathy refers to diseases caused by the immune system losing tolerance to its own antigens, leading to activated immune responses and immune-related pathological damage. These include membranous nephropathy, minimal change disease, focal segmental glomerulosclerosis, IgA nephropathy, and lupus nephropathy. Membranous nephropathy (MN) is an autoimmune glomerular disease characterized by subepithelial deposition of immune complexes in the glomerular basement membrane, accompanied by diffuse thickening of the glomerular basement membrane epithelium. Clinical symptoms mainly manifest as nephrotic syndrome-related symptoms, such as massive proteinuria, hypoalbuminemia, severe edema, and hyperlipidemia. Based on etiology, it can be divided into idiopathic membranous nephropathy and secondary membranous nephropathy. It commonly affects individuals over 40 years of age, and the onset of membranous nephropathy is often insidious. Approximately one-third of patients develop end-stage renal failure within ten years. In recent years, the incidence of membranous nephropathy has been gradually increasing, posing a serious threat to people's health and economic well-being.

[0003] Currently, modern medicine mainly uses immunosuppressants to treat membranous nephropathy, with the goal of exhausting pathogenic immune cells. For example, biological agents targeting CD20 are used to exhaust B cells, or calcineurin inhibitors (CNIs) are used to kill T cells, or cyclophosphamide is used when the former two are ineffective, which simultaneously kills T cells and B cells and other proliferating cells (Membranous nephropathy[J]. J Bras Nefrol. 2023;45(2): 229-243.). However, although immunosuppressive therapy can achieve clinical remission in some patients, the following problems still exist: (1) Some patients are difficult to achieve remission, and about 20%–30% of patients do not respond well to first-line treatment; (2) The relapse rate is high, with a relapse rate as high as 40% after CNIs are discontinued; (3) The safety is low, and long-term immunosuppression significantly increases the risk of adverse reactions such as serious infection, bone marrow suppression and metabolic disorders (Novel Treatments Paradigms: MembranousNephropathy[J]. Kidney Int Rep. 2023;8(3):419-431.). Therefore, exploring safer and more effective treatment options is an urgent clinical problem to be solved in the treatment of membranous nephropathy.

[0004] In addition, minimal change disease is also a major pathological type of autoimmune nephropathy, accounting for 10% to 15% of adult nephrotic syndrome patients and is a common cause of nephropathy in children. Minimal change disease can lead to glomerular podocyte damage and increased glomerular permeability, characterized by no significant changes in glomerular appearance under light microscopy, no immune complex deposition under fluorescence, and diffuse disappearance of podocyte foot processes under electron microscopy. Western medicine treatment for minimal change disease is mainly based on hormones. High-dose oral glucocorticoids are the initial treatment for MCD. Although some patients are sensitive to hormone therapy, some patients are not sensitive to hormones or frequently relapse after remission. Immunosuppressants such as cyclophosphamide, calcineurin inhibitors, and mycophenolate mofetil are alternative drugs for patients who are not sensitive to hormone therapy, mainly by killing immune cells and promoting podocyte repair. In recent years, studies have found that rituximab can also achieve certain effects, but there are few clinical studies on it. In addition, prednisone or prednisolone is the main drug for steroid treatment during disease flare-ups. However, there is no clear consensus on the optimal dosage and duration of treatment for this drug (Minimal Change Disease. Clin J Am Soc Nephrol. 2017 Feb 7;12(2):332-345.).

[0005] According to clinical manifestations, membranous nephropathy and minimal change disease, etc., can be classified into the category of "edema disease" or "water-qi disease" in traditional Chinese medicine, which are closely related to the lungs, spleen, and kidneys. However, according to the immunological pathogenesis, autoimmune nephropathy can also be classified as a type of visceral rheumatism, that is, when the body is exposed to wind, cold and dampness, the body's vital energy is insufficient and latent pathogens reside in the internal organs, and the latent pathogens can be released internally, leading to the onset of the disease. In previous practice and research, the inventors have continuously optimized the traditional Chinese medicine treatment plan for autoimmune nephropathy, including membranous nephropathy and minimal change disease, striving to partially replace the efficacy of immunosuppressants. Based on the Six Channels theory, the inventors' team previously disclosed a traditional Chinese medicine composition for treating membranous nephropathy in patent CN108686188B (hereinafter referred to as patent 1), mainly targeting proteinuria in membranous nephropathy; in order to improve clinical efficacy, the formulation was optimized based on patent 1, and a traditional Chinese medicine composition and preparation method for treating kidney disease were disclosed in patent CN112717111B (hereinafter referred to as patent 2), focusing on edema of yin syndrome in kidney disease patients. However, with clinical diagnosis and continuous research, a deeper understanding has been gained of the treatment of autoimmune nephropathy with traditional Chinese medicine, especially regarding the effects of harmonizing the body's vital energy and protecting the spleen and stomach on the treatment of membranous nephropathy and minimal nephropathy.The significance of this invention for autoimmune nephropathy such as membranous nephropathy, leading to the development of superior and more suitable traditional Chinese medicine prescriptions.

[0006] Based on long-term clinical practice and modern pharmacology, this invention proposes a traditional Chinese medicine composition targeting the core pathogenesis of membranous nephropathy and minimal change disease: "spleen and kidney yang deficiency, internal wind-dampness, and water retention." Clinical observation and in vitro / in vivo experiments have shown that this traditional Chinese medicine composition is more effective in treating membranous nephropathy and minimal change disease than existing traditional Chinese medicine prescriptions, and no significant side effects have been observed in clinical application. Based on this, this invention is completed.

[0007] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating kidney disease, the traditional Chinese medicine composition comprising the following traditional Chinese medicines in parts by weight: 3-30 parts ephedra, 5-120 parts astragalus, 3-30 parts prepared aconite root, 5-60 parts poria cocos, 5-50 parts cinnamon twig, 5-60 parts stir-fried atractylodes macrocephala, 5-60 parts bupleurum root powder, 3-50 parts prepared licorice root, 3-50 parts fresh ginger, and 3-50 parts jujube. The traditional Chinese medicine composition can significantly improve edema, massive proteinuria, and hypoalbuminemia, and at the same time has significant therapeutic effects on symptoms such as abdominal pain and diarrhea, and has anti-inflammatory effects.

[0008] Preferably, the traditional Chinese medicine composition comprises the following parts by weight of traditional Chinese medicine: 5-25 parts ephedra, 15-100 parts astragalus, 5-25 parts prepared aconite root, 10-50 parts poria cocos, 10-30 parts cinnamon twig, 10-30 parts stir-fried atractylodes macrocephala, 8-30 parts bupleurum root, 8-30 parts prepared licorice root, 5-40 parts fresh ginger, and 5-40 parts jujube.

[0009] More preferably, the traditional Chinese medicine composition comprises the following parts by weight of traditional Chinese medicine: 6-15 parts ephedra, 20-60 parts astragalus, 8-20 parts prepared aconite root, 15-30 parts poria cocos, 10-20 parts cinnamon twig, 15-30 parts stir-fried atractylodes macrocephala, 9-25 parts bupleurum root, 6-15 parts prepared licorice root, 9-15 parts fresh ginger, and 9-15 parts jujube.

[0010] Further, in the traditional Chinese medicine composition, the principal herbs are: ephedra and black aconite root slices; the assistant herbs are: astragalus, cinnamon twig, poria, and fangji; the adjuvant herbs are: stir-fried atractylodes macrocephala, ginger, and jujube; and the guiding herb is prepared licorice root.

[0011] Further, the traditional Chinese medicine composition can be modified by adding 1-2 medicinal and edible herbs according to specific symptoms, provided that the core formula remains unchanged. For example, if sleep is poor, albizia bark or jujube seed can be added as an adjuvant; if constipation is present, hemp seed or rhubarb can be added; if blood stasis is present, notoginseng powder, turmeric, or leech can be added; if sweating is significant, ginseng or white peony root can be added.

[0012] Even further, the core formula consists of ephedra, black aconite root slices, astragalus, poria, cinnamon twig, powdered fangji, stir-fried atractylodes macrocephala, ginger, jujube, and prepared licorice root.

[0013] Furthermore, the medicinal and edible herbs are preferably one or more of the following: coix seed, fox nut, yam, and / or red adzuki bean.

[0014] In a second aspect, the present invention provides a traditional Chinese medicine preparation for treating kidney disease, the preparation comprising the following components:3-30 parts ephedra, 3-30 parts black aconite root slices, 5-120 parts astragalus, 5-60 parts poria, 5-50 parts cinnamon twig, 5-60 parts fangji powder, 5-60 parts stir-fried atractylodes macrocephala, 3-50 parts ginger, 3-50 parts jujube, 3-50 parts prepared licorice root, and excipients required for preparing the corresponding traditional Chinese medicine dosage form. Further, the traditional Chinese medicine dosage form is one of the following: decoction, tablet, pill, capsule, granule, powder, oral liquid, or traditional Chinese medicine compound.

[0015] Further, the excipients include fillers, binders, disintegrants, lubricants, and flavoring agents.

[0016] In a third aspect, the present invention provides a method for preparing a traditional Chinese medicine composition for treating kidney disease, wherein the traditional Chinese medicine composition is obtained by the following preparation method: Preparation method 1 includes the following steps: S01, take the prescribed amount of black aconite slices, soak them in water, adjust the pH, decoct and extract, ultrafilter the extract through an ultrafiltration membrane, concentrate and dry to obtain powder A; S02, take the prescribed amount of ephedra and fangji, pulverize them, soak them in water, adjust the pH, extract them by ultrasonication, concentrate and dry to obtain powder B; S03, take the prescribed amount of ginger, wash and juice it, filter it to obtain ginger juice, and set it aside; S04, take the prescribed amount of astragalus, poria, cinnamon twig, stir-fried atractylodes macrocephala, prepared licorice and jujube, decoct them in water, filter them, concentrate them to a clear paste, dry them to obtain powder C; S05, mix the powders A, B and C obtained in steps S01, S02 and S04, add the ginger juice obtained in step S03, mix them, dry them, pulverize them to obtain the traditional Chinese medicine composition.

[0017] Preparation method 2 includes the following steps: S01, take the prescribed amount of black aconite slices, ephedra and fangji powder, pulverize and mix, soak in water, adjust pH, extract with ultrasound to obtain extract, concentrate and dry to obtain powder A; S02, take the prescribed amount of ginger, wash and juice, filter to obtain ginger juice, set aside; S03, take the prescribed amount of astragalus, poria, cinnamon twig, stir-fried atractylodes macrocephala, prepared licorice and jujube, decoct in water, filter and concentrate to clear paste, dry to obtain powder B; S04, mix powder A and powder B from steps S01 and S03 evenly, add ginger juice obtained in step S02, mix evenly, dry, pulverize to obtain traditional Chinese medicine composition.

[0018] Fourth aspect, the present invention provides the application of the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine preparation of the second aspect in the preparation of a drug for treating immune nephropathy.

[0019] Further, the immune nephropathy includes primary immune nephropathy and secondary immune nephropathy.

[0020] Furthermore, the primary immune nephropathy includes membranous nephropathy, minimal change disease, IgA nephropathy, focal segmental glomerulosclerosis, mesangial proliferative glomerulonephritis, and membranoproliferative glomerulonephritis.

[0021] Preferably, the primary immune nephropathy includes membranous nephropathy and minimal change disease.

[0022] Furthermore, the traditional Chinese medicine diagnosis of membranous nephropathy is spleen and kidney yang deficiency with water retention; the diagnosis of minimal change disease is...Traditional Chinese medicine diagnosis is spleen and kidney yang deficiency with water retention.

[0023] The medicinal properties and effects of each Chinese medicinal material in this invention: Ephedra: pungent and slightly bitter in taste, warm in nature. It enters the lung and bladder meridians. It has the effects of inducing sweating and dispelling cold, clearing the lungs and relieving asthma, and promoting diuresis and reducing swelling. It is used for wind-cold common cold, chest tightness and cough, and edema.

[0024] Aconitum carmichaelii: pungent and sweet in taste, very hot in nature; toxic. It enters the heart, kidney, and spleen meridians. It has the effects of restoring yang and rescuing from collapse, tonifying fire and assisting yang, and dispelling cold and relieving pain. It is used for collapse due to yang deficiency, cold limbs and weak pulse, insufficient heart yang, chest pain, vomiting and diarrhea due to deficiency and cold, cold pain in the stomach and abdomen, kidney yang deficiency, impotence and cold uterus, edema due to yin and cold, yang deficiency and external invasion, and cold-damp bi syndrome.

[0025] Astragalus membranaceus: slightly warm in nature, sweet in taste, enters the spleen and lung meridians. It has the effects of invigorating qi and raising yang, consolidating the exterior and stopping sweating, promoting diuresis and reducing swelling, generating fluids and nourishing blood, and detoxifying and draining pus. Used for chronic diarrhea, rectal prolapse, uterine prolapse, excessive sweating, urticaria, edema, dizziness, palpitations, and slow-healing sores caused by deficiency of Qi.

[0026] Poria: sweet, bland, and neutral in nature, and enters the heart, lung, spleen, and kidney meridians. It has the effects of promoting diuresis and eliminating dampness, strengthening the spleen, and calming the mind. Used for spleen deficiency with scanty food, loose stools and diarrhea, restlessness, palpitations and insomnia, edema with scanty urine, phlegm retention and dizziness.

[0027] Cinnamon twig: pungent and sweet in taste, and warm in nature. Enters the heart, lung, and bladder meridians. It has the effects of inducing sweating and relieving muscle tension, warming and unblocking the meridians, assisting Yang and transforming Qi, and calming and descending Qi. Used for wind-cold common cold, cold pain in the stomach and abdomen, blood cold and amenorrhea, joint pain, phlegm retention, edema, palpitations, and running piglet syndrome.

[0028] Fried Atractylodes macrocephala: bitter and sweet in taste, and warm in nature. Enters the spleen and stomach meridians. It has the effects of strengthening the spleen and replenishing qi, drying dampness and promoting diuresis, stopping sweating, and calming the fetus. It is used for spleen deficiency with poor appetite, abdominal distension and diarrhea, phlegm retention and dizziness, edema, spontaneous sweating, and threatened abortion.

[0029] Stephania tetrandra: bitter in taste and cold in nature. It enters the bladder and lung meridians. It has the effects of dispelling wind and relieving pain, promoting diuresis and reducing swelling. It is used for rheumatic pain, edema and beriberi, difficulty in urination, eczema and sores.

[0030] Ginger: pungent in taste and slightly warm in nature. It enters the lung, spleen, and stomach meridians. It has the effects of relieving exterior syndromes and dispersing cold, warming the middle and stopping vomiting, resolving phlegm and stopping cough, and detoxifying fish and crab poisoning. It is used for wind-cold common cold, stomach cold vomiting, cold phlegm cough, and fish and crab poisoning.

[0031] Jujube: sweet in taste and warm in nature. It enters the spleen, stomach, and heart meridians. It has the effects of replenishing the middle and replenishing qi, nourishing blood and calming the mind. It is used for spleen deficiency with poor appetite, fatigue and loose stools, and hysteria in women.

[0032] Prepared licorice root: sweet in taste and neutral in nature. It enters the heart, lung, spleen, and stomach meridians. It has the effects of tonifying the spleen and stomach, replenishing qi and restoring pulse. It is used for spleen and stomach weakness, fatigue, palpitations, and irregular pulse.

[0033] The treatment principle in this invention: warming yang and resolving stagnation, relieving exterior symptoms and expelling pathogens.

[0034] This invention is the result of combining long-term clinical practice with modern pharmacological research. The treatment principle is based on "warming yang and resolving stagnation, relieving exterior symptoms and expelling pathogens" as the core principle. This principle focuses on treating complex edema syndrome of "spleen and kidney yang deficiency, wind-dampness, and water retention". The principal drugs are ephedra and black aconite.

[0035] Ephedra: relieves exterior symptoms and dispels cold, promotes lung function and diuresis; black aconite: warms and tonifies kidney yang, resolves stagnation and eliminates stagnation. The two drugs are the principal drugs, and black aconite warms yang.Ephedra is used to dispel pathogens and strengthen the body's resistance. The two herbs are used together for the symptoms of yang deficiency and accumulation, internal stagnation of pathogens, and overflow of water retention in this disease.

[0036] Assistant herbs: Astragalus, Cinnamon Twig, Poria, and Stephania.

[0037] Astragalus: Strengthens the stomach and consolidates the exterior, promotes water metabolism and reduces swelling. It tonifies the stomach qi, and when the stomach qi is sufficient, water flows smoothly, promoting the circulation of body fluids, promoting diuresis and reducing swelling; at the same time, Astragalus assists the principal herb in tonifying the body's resistance and assists in expelling pathogens. Cinnamon Twig: Warms and unblocks yang qi, regulates the four limbs, and transforms water and dampness. It assists Aconitum in warming and unblocking, and when combined with Astragalus, it tonifies qi and unblocks yang, relieves muscle tension and disperses pathogens. Poria: An essential herb for promoting diuresis and eliminating dampness, when combined with Cinnamon Twig, it warms and transforms water and dampness between the skin. Stephania: Dispels wind and dampness, promotes diuresis and reduces swelling, when combined with Astragalus, it expels the pathogenic water and dampness that lingers in the meridians of the skin, and at the same time restrains the heat of the herbs. The assistant herbs target the stomach deficiency and the inability to transform body fluids, and the retention of dampness. They assist the principal herbs in warming the yang and benefiting the stomach, promoting the distribution of body fluids, transforming qi and promoting diuresis, and expelling pathogens.

[0038] The adjuvant herbs are: stir-fried Atractylodes macrocephala, ginger and jujube.

[0039] Stir-fried Atractylodes macrocephala: strengthens the spleen and dries dampness. It helps Astragalus membranaceus to strengthen the spleen and benefit the stomach, dry dampness and consolidate the interior, and also helps Cinnamomum cassia and Poria cocos to warm the middle and transform qi, so that the body has no source of dampness. Ginger and jujube: strengthen the spleen and harmonize the stomach, and disperse dampness. They can help Astragalus membranaceus and Cinnamomum cassia to warm the yang and release the exterior, and consolidate the stomach qi, and also harmonize the herbs. The adjuvant herbs strengthen the spleen to eliminate the source of dampness on the one hand, and benefit qi and blood and replenish the body's vital energy on the other hand, and help the principal and assistant herbs to perform their functions.

[0040] The guiding herb is: roasted licorice.

[0041] Roasted licorice: moderates the properties of herbs (such as ephedra and aconite), and can be used with other herbs to benefit qi and harmonize the middle, and transform yang with pungent and sweet herbs.

[0042] The traditional Chinese medicine composition of the present invention has a strict combination of principal, assistant, adjuvant, and guide herbs, and regulates the lung, spleen, and kidney organs in a coordinated manner. It integrates the methods of clearing the lungs, strengthening the spleen, warming the kidneys, promoting diuresis, and strengthening the defensive qi into one, so as to achieve the effects of clearing the lungs and promoting diuresis, strengthening the spleen and warming the kidneys, strengthening the defensive qi and reducing swelling. It hits the pathogenesis and is an innovative core effective prescription for the treatment of immune nephropathy with traditional Chinese medicine compound.

[0043] Beneficial effects The present invention adopts the method of warming yang and relieving exterior syndrome, so as to achieve the effects of clearing the lungs and promoting diuresis, strengthening the spleen and warming the kidneys, strengthening the defensive qi and reducing swelling, and treating immune-related nephropathy, especially membranous nephropathy and minimal change disease.

[0044] The drug composition of the present invention is a pure traditional Chinese medicine preparation. Clinical observation and in vitro and in vivo basic tests show that the prescription can significantly improve edema, massive proteinuria and hypoalbuminemia, etc., and has better effects on symptoms such as abdominal pain and diarrhea than patent 2. It also has anti-inflammatory effects, and no obvious side effects have been found in clinical application. The treatment is particularly effective for patients with membranous nephropathy and minimal change disease diagnosed by traditional Chinese medicine as having spleen and kidney yang deficiency, internal wind-dampness, and water retention.

[0045] Figure 1 shows the biochemical indicators of each group of rats.

[0046] Note: A represents the 24-hour urinary protein quantification (24hUTP) results of each group of rats during the 12-week treatment period; B represents the serum albumin (ALB) level of each group of rats; C represents the serum creatinine (SCr) level of each group of rats; D represents the serum cholesterol (CHO) level of each group of rats; E represents...Triglyceride (TG) levels in each group of rats; F represents low-density cholesterol ester (LDL-C) levels in each group of rats; G represents blood urea nitrogen (BUN) levels in each group of rats; H represents alanine aminotransferase (ALT) levels in each group of rats; I represents aspartate aminotransferase (AST) levels in each group of rats. Data are expressed as mean ± standard deviation. *P<0.05, **P<0.01, ***P<0.001 indicate statistically significant differences between the two groups.

[0047] Figure 2 shows the pathological damage to the kidneys of PHN rats after intervention.

[0048] Note: Pathological images (×400) and images of the glomerular filtration barrier (×3000k) observed under a light microscope and a transmission electron microscope are shown in the figures. Black arrows in the figures indicate electron-dense deposits on the basement membrane. Red arrows in the figures indicate podocytes with reduced edema after traditional Chinese medicine treatment.

[0049] Figure 3 shows the inhibitory level of complement activation in PHN rats in each group.

[0050] Note: A represents the serum C3a level of each group of rats; B represents the serum C5a level of each group of rats (data are expressed as mean ± standard deviation. *P<0.05 indicates that the difference between the two groups is statistically significant); C represents the immunohistochemical staining images of C5b-9 deposition in the glomeruli of each group of rats (×400); D represents the immunofluorescence staining images of C3 deposition in the glomeruli of each group of rats (×400).

[0051] Figure 4 shows the overall remission rate and the number of patients who received remission in the two groups of patients.

[0052] Figure 5 shows the comparison of the efficacy of single syndromes in traditional Chinese medicine between the two groups of patients after treatment. Detailed Embodiments

[0053] The specific embodiments of the present invention will be further described below. It should be noted that the description of these embodiments is for the purpose of helping to understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.

[0054] Unless otherwise specified, the experimental methods in the following embodiments are conventional methods, and the experimental materials used in the following embodiments can be purchased through conventional commercial channels unless otherwise specified.

[0055] Example 1 Traditional Chinese Medicine Composition and its Preparation Components of the Traditional Chinese Medicine Composition: Ephedra 10g, Aconite 15g, Astragalus 30g, Poria 15g, Cinnamon Twig 10g, Stephania tetrandra 10g, Atractylodes macrocephala 12g, Ginger 10g, Jujube 15g, Licorice Root 10g.

[0056] The traditional Chinese medicine composition was prepared by the following method.

[0057] Preparation method 1 includes the following steps: S01, take the prescribed amount of Aconite, add 1-2 times the amount of water and soak for 3-4 hours, adjust the pH value to 3-5, decoct and extract for 2-4 hours, use an ultrafiltration membrane with a molecular weight cutoff of 5KD-10KD to obtain the extract, concentrate and dry to obtain powder A;S02, take the prescribed amount of ephedra and fangji, grind them into coarse powder, add 1-2 times the amount of water to soak, adjust the pH value to 3-5, soak at room temperature for 3-6 hours, ultrasonically extract for 2-4 hours, extract at 50-75℃, maintain pH value, extract 1-2 times, combine the extracts, concentrate and dry to obtain powder B; S03, take the prescribed amount of ginger, wash and juice, filter to obtain ginger juice, set aside; S04, take the prescribed amount of astragalus, poria, cinnamon twig, stir-fried atractylodes macrocephala, prepared licorice and jujube, add 8-12 times the amount of water to decoct for 1-2 hours, twice in total, combine the decoctions, filter, concentrate to a clear extract with a relative density of 1.10-1.20 (measured at 60℃), dry to obtain powder C; S05, mix the powders A, B and C obtained in steps S01, S02 and S04, add the ginger juice obtained in step S03, mix well, dry, grind to obtain the traditional Chinese medicine composition.

[0058] Preparation method 2 includes the following steps: S01, take the prescribed amount of black aconite slices, ephedra and fangji powder, crush them into coarse powder, mix them, add 1-2 times the amount of water to soak, adjust the pH value to 3-5, soak at room temperature for 3-6 hours, ultrasonically extract for 2-4 hours, extract at 50-75℃, maintain the pH value, extract 1-2 times, combine the extracts, concentrate and dry to obtain powder A; S02, take the prescribed amount of ginger, wash and juice it, filter to obtain ginger juice, and set aside; S03, take the prescribed amount of astragalus, poria, cinnamon twig, stir-fried atractylodes macrocephala, prepared licorice and jujube, add 8-12 times the amount of water to decoct for 1-2 hours, twice in total, combine the decoctions, filter, concentrate to a clear paste with a relative density of 1.10-1.20 (measured at 60℃), dry to obtain powder B; S04, mix powder A and powder B from steps S01 and S03 evenly, add ginger juice obtained in step S02, mix well, dry, and pulverize to obtain a traditional Chinese medicine composition.

[0059] Add the excipients required for preparing the corresponding traditional Chinese medicine dosage form to the traditional Chinese medicine compositions obtained by the above preparation methods 1 and 2 to obtain the traditional Chinese medicine preparation.

[0060] Example 2 Clinical efficacy and safety observation 1. Clinical data 1.1 Case source This study takes patients with membranous nephropathy diagnosed by traditional Chinese medicine as spleen and kidney yang deficiency and water and dampness overflow as the research subjects. The included cases are patients who visited the Department of Nephrology of Beijing Hospital of Traditional Chinese Medicine, Capital Medical University from January 2023 to June 2024 and were selected as qualified by the inclusion and exclusion criteria.

[0061] 1.2 Diagnostic Criteria The Western medicine diagnostic criteria refer to the 4th edition of *Nephrology*, namely: (1) patients with primary membranous nephropathy diagnosed by renal biopsy; (2) patients whose renal biopsy confirms atypical membranous nephropathy and excludes secondary factors such as systemic lupus erythematosus; (3) patients who are positive for anti-PLA2R1 antibody by serological testing and whose clinical manifestations are consistent with primary membranous nephropathy and exclude secondary factors.

[0062] The TCM diagnostic and syndrome differentiation criteria are formulated with reference to the standards in *Internal Medicine of Traditional Chinese Medicine* edited by Zhou Zhongying.

[0063] Main symptoms: chills, cold hands and feet, soreness in the lower back, edema, abdominal distension, abdominal pain, loose stools or dry stools, pale and tender tongue, thin and greasy or slippery tongue coating, deep pulse.

[0064] Secondary symptoms: fatigue, shortness of breath, reluctance to speak, spontaneous sweating, scanty urination, frequent urination at night, etc.

[0065] If the patient has the above three main symptoms and two secondary symptoms, the patient can be diagnosed with spleen and kidney yang deficiency and water retention syndrome. Instructions for use 6 / 14 pages 9 CN 121401392 A

[0066] 1.3 Inclusion criteria: age 18-80 years; gender not limited; patients with a confirmed diagnosis of membranous nephropathy by renal biopsy or patients with positive serum antiphospholipase A2 antibody; clinical manifestations that meet the criteria for low, medium and high risk patients as defined by the Kidney Disease: Improving Global Outcomes (KDIGO) guidelines (see table below); patients who voluntarily accept simple traditional Chinese medicine treatment.

[0067] 1.4 Exclusion Criteria: Secondary MN; Patients with other renal impairments; Patients allergic to Chinese medicine components; Patients with severe cardiovascular and cerebrovascular diseases, cancer, or other serious diseases; Patients with severe psychological or mental abnormalities; Patients who are not regularly followed up or lost to follow-up.

[0068] 1.5 Dropout Criteria: Patients who are unsuitable to continue the trial due to adverse events or whose doctors deem them unsuitable to continue the trial; Patients who develop certain serious comorbidities or complications, experience pregnancy, die, or are lost to follow-up; Patients who use other prohibited treatments or drugs, affecting the determination of efficacy and safety.

[0069] 2. Research Methods 2.1 Treatment Methods: All enrolled patients received standardized prescription treatment. The core prescription consisted of Ephedra 10g, Aconite 15g, Astragalus 30g, Poria 15g, Cinnamon Twig 10g, Stephania Tetrandra 10g, Atractylodes Macrocephala 12g, Ginger 10g, Jujube 15g, and Prepared Licorice Root 10g. The traditional decoction is administered orally twice daily, 200 mL each time.

[0070] Under the premise of keeping the core formula unchanged, the attending physician is allowed to add 1-2 Chinese medicinal herbs (including coix seed, euryale seed, yam, and red adzuki bean) according to the patient's specific symptoms. The standard treatment course is 12 months. For those who do not achieve remission, the treatment can be extended. For those who achieve complete remission, a dose reduction maintenance regimen is initiated.

[0071] All patients simultaneously receive optimized supportive care based on the KDIGO guidelines, including: a high-quality, low-protein, low-phosphorus, and adequate-calorie diet; angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists to control blood pressure at 120-135 / 75-85 mmHg; symptomatic diuresis to correct water, electrolyte, and acid-base imbalances; anti-infective treatment for those with infections; and routine rivaroxaban or warfarin anticoagulation therapy for serum albumin below 25 g / L.

[0072] 2.2 Observation Indicators 2.2.1 Laboratory Indicators(1) Primary efficacy indicators: 24-hour urine protein quantification (24hUTP), serum albumin (ALB).

[0073] (2) Secondary efficacy indicators: Renal function: including serum creatinine (SCr), estimated glomerular filtration rate (eGFR); blood lipid status: including serum total cholesterol (TCHO), serum triglycerides (TG), serum low-density lipoprotein (LDL-C). eGFR was calculated using the Xiangya formula, and the calculation formula is as follows: eGFR (ml / (min*1.73m2))=186× (Scr)^-1.154× (age)^-0.203× (0.742 female).

[0074] Safety indicators: General safety indicators: liver function, kidney function, etc.

[0075] 2.2.2 TCM syndrome score The main observed symptoms are aversion to cold, cold hands and feet, soreness in the lower back, edema, abdominal distension and pain, loose stools or dry stools, fatigue, shortness of breath and reluctance to speak, spontaneous sweating, and frequent urination. The above symptoms are divided into four levels according to severity: 0 points for no symptoms, 1 point for mild, 2 points for moderate, and 3 points for severe. The scores are counted once before and after treatment.

[0076] 2.2.3 Clinical efficacy Clinical remission rate calculation: The clinical condition is judged according to the level of serum albumin and 24-hour urine protein quantification. Clinical remission rate = ((number of complete remissions + number of partial remissions) / total number of cases in this group) × 100%.

[0077] (1) Complete remission criteria: 24-hour urine protein quantification ≤ 0.3g, serum albumin > 35g / L, and eGFR > 60 [mL / (min·1.73m2)].

[0078] (2) Partial relief criteria: 24-hour urine protein quantification ≤3.5g, and a decrease of ≥50% from baseline; serum albumin level rises or returns to normal; serum creatinine level remains stable.

[0079] (3) No relief criteria: 24-hour urine protein quantification decreases <50% from baseline, or 24-hour urine protein quantification >3.5g.

[0080] 2.2.4 The efficacy of TCM syndromes is formulated with reference to the "Guiding Principles for Clinical Research of New Chinese Medicines (Trial)": Clinical control: TCM clinical symptoms and signs disappear or basically disappear, and the syndrome efficacy rate is >90%; Significant effect: TCM clinical symptoms and signs are significantly improved, and the syndrome efficacy rate is >70% (<90%); Effective: TCM clinical symptoms and signs are all improved, and the syndrome efficacy rate is >30% (<70%); Ineffective: TCM clinical symptoms and signs are not significantly improved, or even aggravated, and the syndrome efficacy rate is <30%. (Syndrome efficacy rate (nimodipine method) = [(total score before treatment - total score after treatment) ÷ total score before treatment] x 100%).

[0081] 2.3 Sample size calculation This invention is based on clinical series case observation, with a sample size of 105 cases, according to "Rituxima b or Cyclophosphamide in the Treatment of MembranousThe report "Nephropathy: The RI-CYCLO Randomized Trial" demonstrates clinical efficacy and safety.

[0082] 2.4 Statistical Analysis Methods Instructions 8 / 14 pages 11 CN 121401392 A SPSS 25.0 software was used for data statistical processing. Normally distributed measurement data were expressed as mean ± standard deviation; non-normally distributed measurement data were expressed as median (interquartile range); and count data were expressed as rates. Normally distributed measurement data were statistically analyzed using t-tests; intergroup comparisons were performed using independent samples t-tests; intragroup comparisons before and after treatment were performed using paired t-tests; non-normally distributed or unequal variance measurement data were statistically analyzed using rank-sum tests; two-way ANOVA for non-normally distributed or unequal variance measurement data was performed using the rank-based Scheirer-Ray-Hare test; count data were statistically analyzed using chi-square tests; and ordinal data were analyzed using non-parametric statistical methods (rank-sum tests). P < 0.05 indicates statistical significance.

[0083] Results 3.1 Baseline Data This cohort included 105 patients who met the inclusion and exclusion criteria, with a mean age of 47.5 ± 12.3 years and a median disease duration of 10.0 (4.5, 26.0) months. The cohort included 61 male patients (58.1%) and 44 female patients (41.9%). Among them, 45 patients (42.9%) were high-risk, 34 patients (32.4%) were intermediate-risk, and 26 patients (24.7%) were low-risk. The baseline 24-hour urinary protein quantification for the entire population was 6.0 (2.7, 8.9) g / 24h, serum albumin was 28.7 ± 6.1 g / L, and median serum creatinine was 69.7 (55.7, 89.0) μmol / L (see Table 1).

[0084] Table 1 Patient baseline characteristics 3.2 Clinical efficacy During the treatment, a total of 77 patients (73.3%) achieved clinical remission, of which 49 cases achieved partial remission, accounting for 46.7% of the total number of patients, and 28 cases achieved complete remission, accounting for 26.7% of the total number of patients. (See Table 2) Table 2 Overall remission rate and number of patients with remission 3.3 Analysis of changes in laboratory indicators before and after treatment After treatment, the changes in laboratory indicators of patients were as follows: the average 24-hour urinary protein decreased to 1.6 (0.6, 3.7) g / 24 h, and the average serum albumin increased to 39.4 (33.8, 42.5) g / L. There were statistically significant differences in both before and after treatment (P<0.05). The mean serum creatinine level was 67.2 (52.1, 87.1) μmmol / L, and the mean estimated glomerular filtration rate was 89.2 ± 15.1 mL / (min·1.73m2). No statistically significant difference was observed between the two groups before and after treatment (P>0.05). The mean alanine aminotransferase level was 15.8.The mean values ​​of aspartate aminotransferase (AST) were (11.9, 24.3) U / L and (17.2, 24.5) U / L, with no statistically significant difference before and after treatment (P>0.05) (see Table 3). Furthermore, the positive rate of anti-PLA2R antibodies was 55% before treatment, decreasing to 15% after one year of treatment.

[0085] Table 3 Comparison of laboratory indicators before and after treatment in the overall patient population 3.4 Changes in TCM syndrome scores before and after treatment The TCM syndrome scores of patients decreased to 7.2±5.3 after treatment, showing a statistically significant difference before and after treatment (see Table 4).

[0086] Table 4 Comparison of TCM syndrome scores before and after treatment in the overall patient population 3.5 Safety During treatment and follow-up, no patients experienced adverse events such as respiratory infection, diarrhea, nausea, vomiting, hair loss, or leukopenia. No participants experienced end-stage renal disease (ESKD) or serious adverse events. The main drugs in the traditional Chinese medicine composition provided by this invention include ephedra and aconite, which are known to cause arrhythmia, excessive sweating, shortness of breath and weakness, nausea and vomiting, numbness in the limbs, and difficulty urinating. In the limited clinical data, only one patient withdrew from treatment due to stomach discomfort after taking the medicine, three patients experienced difficulty urinating, and the symptoms were relieved after the dosage was reduced. No adverse drug reactions were observed in the remaining patients.

[0087] Example 3 Clinical comparative observation of different formulations The intermediate and high-risk patients in Example 1 were selected and compared with the cohort of patients with membranous nephropathy treated. The patients were matched according to baseline age, sex, serum albumin level, proteinuria, serum creatinine and eGFR. Each group included 57 patients. Instructions for Use, Pages 10 / 14, 13 CN 121401392 A

[0088] Under the same basic treatment in the research plan, the treatment group is the Chinese medicine composition described in this invention, and the control group is the Chinese medicine composition described in Patent 2: (1) Basic treatment: high-quality low-protein, low-phosphorus, and sufficient calorie diet; angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists to control blood pressure at 120~135 / 75~85 mmHg; symptomatic diuresis to correct water, electrolyte and acid-base balance; anti-infective treatment for those with infection; routine anticoagulation treatment with rivaroxaban or warfarin when serum albumin is below 25 g / L.

[0089] (2) The treatment group is the Chinese medicine composition described in this invention (see Example 1): Ephedra 10g, Astragalus 30g, Aconitum carmichaelii 15g, Poria cocos 15g, Cinnamomum cassia 10g, Atractylodes macrocephala 12g, Stephania tetrandra 10g, Glycyrrhiza uralensis 10g, Zingiber officinale 10g, Ziziphus jujuba 15g.

[0090] (3) The herbal composition of the control group was: 10g of raw ephedra, 12g of black aconite root slices, 30g of raw astragalus root, 15g of poria cocos, 10g of fangji (Stephania tetrandra), 10g of ginger, 10g of alisma plantago-aquatica, 10g of cinnamon twig, and 6g of prepared licorice root.

[0091] The herbal composition of the control group was decocted with water for 24 hours, twice, the decoction was concentrated, dried and powdered, and then added to the decoction.Pharmaceutically acceptable carriers and / or excipients.

[0092] Observation indicators and statistical analysis The observation indicators and statistical analysis methods in this embodiment refer to the clinical study described in Example 2.

[0093] Results 3.1 Baseline characteristics of the two groups of patients The baseline characteristics of the two groups of patients are shown in Table 5. There was no statistically significant difference in general and disease conditions between the two groups.

[0094] Table 5 Baseline characteristics of patients 3.2 Comparison of efficacy After 6 months and 12 months of treatment, the remission rate of both groups increased significantly. After 12 months of treatment, the remission rate of both groups increased significantly, but the difference between the two groups was small (see Figure 4).

[0095] Based on the above results and clinical treatment experience, 51 patients in each group were matched for a treatment follow-up period of 18 months. Statistical analysis showed that as the treatment time increased, the remission rate of both groups increased compared with the previous period, and the remission rate of the treatment group was significantly higher than that of the control group. The difference between the two groups was statistically significant (P=0.02).

[0096] Table 6 Remission status of the two groups of patients after 18 months of follow-up 3.3 Changes in laboratory indicators After 12 months of treatment, the quantitative urinary protein in both groups decreased (P<0.05), and the quantitative albumin increased significantly (P<0.05). There was no difference in serum creatinine and eGFR between the two groups during the treatment process (see Table 7). After 18 months of treatment, the decrease in urinary protein in the treatment group was more significant than that in the control group.

[0097] Table 7 Changes in main laboratory indicators of the two groups Note: *P<0.05 compared with baseline for each group, #P<0.05 compared with the two groups.

[0098] 3.4 Analysis of efficacy of single syndrome in traditional Chinese medicine The total effective rate of the treatment group in improving the three syndromes of fatigue, spontaneous sweating, abdominal distension and abdominal pain was significantly higher than that of the control group (P<0.05) (see Figure 5).

[0099] The inventors' team has conducted long-term clinical practice in the treatment of membranous nephropathy with traditional Chinese medicine. The application of the traditional Chinese medicine composition described in this invention to treat membranous nephropathy can significantly reduce urinary protein excretion and increase serum albumin levels. With the extension of treatment time, the clinical remission rate increases significantly. Liver and kidney function are regularly monitored throughout the treatment process, and no obvious adverse reactions are observed with long-term use. At the same time, this traditional Chinese medicine composition is significantly better than the traditional Chinese medicine composition described in Patent 2 in improving clinical symptoms such as fatigue, shortness of breath, edema, abdominal distension and abdominal pain in patients.

[0100] Example 4 Animal Experiment Comparison Experiment Method Animal Model Construction and Grouping Preparation Passive Heymann Nephritis (PHN) Model. A total of 36 male SD rats aged 6-7 weeks and weighing 130-150g were purchased from Beijing Huafukang Biotechnology Co., Ltd. (License No.: SCXK (Beijing) 2016–0002). All animals were housed in an environment free of specific pathogens and kept at a constant temperature (22±The animals were kept at a constant temperature (2℃) and humidity (55±5%), with alternating light and dark periods for 12 hours, and free access to food and water. After 3 days of acclimatization, 6 out of 36 male SD rats were randomly assigned to the blank control group (CTL), and the CTL received a tail vein injection of 0.5 ml / 100 g physiological saline. The remaining 30 rats received a single tail vein injection of 0.5 ml / 100 g goat anti-rat Fx1A antibody serum (PTX-002S, Probetex, USA). One week after the injection, 24-hour urine protein quantification (24hUTP) was performed. A 24hUTP > 10 mg was considered a successful model. The 30 successfully modeled PHN rats were randomly divided into a model group, a cyclosporine A (CsA) group, a traditional Chinese medicine group 1, a traditional Chinese medicine group 2, and a traditional Chinese medicine group 3.

[0101] Intervention measures Cyclosporine A (CsA) group: Cyclosporine A was purchased from Hangzhou Sino-American East China Pharmaceutical Co., Ltd., production batch number FJB1006004, specification 25 mg / capsule. Cyclosporine was injected into the tail vein, and the rat dose was converted according to body surface area = human clinical dose × 6.3.

[0102] Traditional Chinese medicine group 1: Ephedra 10g, black aconite 15g, stir-fried atractylodes macrocephala 12g, poria cocos 20g, dried ginger 10g and prepared licorice root 10g. Weigh the above traditional Chinese medicine composition according to the prescription. Except for ephedra and aconite, wash and mix the remaining drugs. Add water equivalent to three times the volume of the drugs to the drugs and soak for 30 min. First decoct aconite for 20 min, then add ephedra and decoct for another 10 min. Then add the remaining drugs and heat to decoct. After boiling, continue to heat over low heat and concentrate the decoction to 1 / 3 of the total volume before decoction. After filtering the dregs, the decoction is obtained.

[0103] Group 2 of traditional Chinese medicine: 10g of raw ephedra, 12g of black aconite root slices, 30g of raw astragalus root, 15g of poria cocos, 10g of fangji (Stephania tetrandra), 10g of ginger, 10g of alisma plantago-aquatica, 10g of cinnamon twig, and 6g of prepared licorice root. The above-mentioned traditional Chinese medicine combination was decocted with water for 24 hours, twice. The decoction was concentrated to a certain volume, and the dregs were filtered to obtain a decoction.

[0104] Group 3 of traditional Chinese medicine (see Example 1): 10g of ephedra, 15g of black aconite root slices, 30g of astragalus root, 15g of poria cocos, 10g of cinnamon twig, 12g of stir-fried atractylodes macrocephala, 10g of powdered fangji (Stephania tetrandra), 15g of jujube, 10g of ginger, and 10g of prepared licorice root. The traditional Chinese medicine groups were dissolved in water according to the equivalent dosage and administered orally at 1ml / 100g / d. The CsA dose was 25 mg / kg / d, dissolved in water and administered orally at 1ml / 100g / d. The control group was given 1 ml / 100 g / day of drinking water. This was administered via gavage for 12 consecutive weeks.

[0105] Biochemical indicators were measured every two weeks for 24-hour UTP. 24-hour urine was collected in a metabolic cage and 24-hour UTP was detected by Coomassie Brilliant Blue assay (C035–2-1, Nanjing Jiancheng Biotechnology Institute, China). At the end of the final treatment, after a 12-hour fast, urine was administered via abdominal...Rats were anesthetized by intracavitary injection of 1% sodium pentobarbital, and blood was collected from the abdominal aorta. Blood used for detecting biochemical indicators was collected by a procoagulant serum separation extraction tube, then the serum was separated by centrifugation and stored at −80℃ for detection and analysis.

[0106] Serum biochemical indicators included serum albumin (ALB), serum creatinine (CRE), blood urea nitrogen (BUN), total cholesterol (CHO), triglycerides (TG), low-density lipoprotein (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), which were measured by a Hitachi 7600 automated biochemical analyzer.

[0107] After systemic perfusion with physiological saline, rat kidney tissue was fixed with 4% paraformaldehyde for 2 hours. After dehydration, the tissue was embedded in paraffin and cut into 3 μm thick sections, and stained with hematoxylin and eosin (HE), Masson staining, and periodate hexamine silver staining (PASM)-Masson trichrome staining (P+M). The degree of tubulointerstitial disease in PHN rats was assessed using a semi-quantitative tubulointerstitial scoring system. The specific assessment method was as follows: Each HE-stained kidney section was observed under a ×100x microscope, with 10 fields of view showing tubular atrophy or dilation, interstitial inflammation, and fibrosis. The scoring range was 0 to 3 (0 points, normal tubulointerstitial tissue; 1 point, diseased area <25% of the section; 2 points, diseased area is 25–50% of the section; 3 points, diseased area >50%). The average value was analyzed using Image-J (version 1.48) software to obtain the renal tubulointerstitial disease index.

[0108] Serum complement level detection in rats: Blood from PHN rats was collected using EDTA anticoagulant tubes, plasma was separated by centrifugation, and stored at -80°C for analysis. The concentrations of C3a, C5a, and C5b9 in rat plasma were determined according to the instructions using a commercially available enzyme-linked immunosorbent assay kit.

[0109] PHN Rat Immunohistochemistry and Immunofluorescence Analysis Instructions, Pages 13 / 14, CN 121401392 A. Paraffin-embedded kidney tissue was cut into 3 μm thick sections. After complete dewaxing, antigen retrieval was performed using sodium citrate buffer. The retrieval sections were blocked with 5% bovine serum albumin (BSA), followed by the addition of rabbit anti-C3 antibody (Abcam, catalog number ab200999, USA), and incubated overnight at 4°C. Donkey anti-rabbit IgG (H+L) highly crosslinked secondary antibody (Invitrogen, catalog number A21206, AF488 label) was added to the unlabeled fluorescent antibody. All sections were mounted using anti-fluorescence attenuation mounting medium containing DAPI dye (Solepro, catalog number S2110, China). The immunohistochemical staining procedure was as follows: After pre-cooling the sections at 4°C, they were incubated overnight with rabbit anti-C5b-9 antibody (Abcam, catalog number ab55811, USA). Then use HRPThe labeled secondary antibody (#SP-00223, Bio-Sen Biotechnology, Beijing) was used for a second incubation. Finally, DAB chromogenic agent was added and hematoxylin was stained; brown was a positive staining result. All sections were observed under a confocal microscope (Zeiss LSM 800, Germany).

[0110] To verify the efficacy of the traditional Chinese medicine composition described in this invention in PHN rats, continuous 24-hour UTP measurements were performed. The results are shown in Figure 1A. The urinary protein in PHN rats gradually increased and fluctuated. After 8 weeks, the urinary protein levels in the CsA group and the traditional Chinese medicine group 3 were significantly lower than those in the model group. The urinary protein in the traditional Chinese medicine group 2 was significantly lower than that in the model group at 10 and 12 weeks, while the proteinuria in the traditional Chinese medicine group 1 was significantly lower than that in the model group at 12 weeks. After 12 weeks of intervention, other laboratory indicators of the rats were detected, as shown in Figures 1B-I. Both the traditional Chinese medicine group and the CsA intervention improved the levels of CHO, TG, and LDL-C in the rats. Traditional Chinese medicine (TCM) groups 2 and 3 significantly reduced blood urea nitrogen levels, while TCM group 3 significantly reduced blood creatinine levels. The above experimental results indicate that the TCM composition described in this invention is faster and more effective in treating membranous nephropathy compared to existing formulations.

[0111] The pathological changes in the kidneys of PHN rats are similar to those in membranous nephropathy, characterized by IgG deposition and diffuse thickening of the basement membrane. To verify the effect of TCM group 3 on the pathological damage to the kidneys of PHN rats, this invention observed the pathological changes in the kidneys using an optical microscope (see Figure 2). HE and P+M staining showed that the glomerular basement membrane in the model group was thicker than that in the control group, forming local "nail-like" structures. In contrast, the degree of basement membrane lesions in the cyclosporine group and the TCM group was milder than that in the model group. MASSON staining showed that compared to the control group, the model group and CsA group had more severe renal interstitial fibrosis, while the TCM group had milder renal interstitial damage. This result indicates that TCM treatment has a protective effect on the kidneys of PHN rats.

[0112] The damage to the glomerular filtration barrier in rats was observed by transmission electron microscopy. As shown in Figure 2, compared with the control group, the glomerular basement membrane in the model group was thickened, electron-dense deposits were visible under the epithelial cells, and podocyte processes showed edema and extensive fusion. In contrast, the thickening of the basement membrane in the traditional Chinese medicine group 3 was significantly reduced, and the edema of podocyte processes was also slowed down. This indicates that the traditional Chinese medicine group 3 can effectively reduce podocyte damage and ultrastructural lesions, ultimately repairing the filtration barrier and reducing proteinuria.

[0113] In addition, this invention found that after treatment with the traditional Chinese medicine composition described in this invention, the serum C3a and C5a levels in rats decreased significantly, which was not observed in other traditional Chinese medicine groups (see Figures 3A-B). The histochemical and fluorescence experiments of rat kidney tissue also showed that the level of kidney complement activation was significantly inhibited under the intervention treatment of the traditional Chinese medicine composition described in this invention (see Figures 3C-D).

[0114] The above experimental results further confirm that the traditional Chinese medicine composition described in this invention is effective in treating membranous nephropathy and minimal change disease.It exhibits outstanding efficacy in treating kidney diseases, demonstrating significant therapeutic effects and a clear protective effect on the kidneys. Its therapeutic effects are not only reflected in reducing proteinuria and improving biochemical indicators, but more importantly, it can significantly reduce pathological damage and ultrastructural lesions in the kidneys, and uniquely inhibit the excessive activation of the complement system. Specification 14 / 14 page 17 CN 121401392 A Figure 1 Figure 2 Description description page 1 / 3 page 18 CN 121401392 A Figure 3 Figure 4 Description description page 2 / 3 page 19 CN 121401392 A Figure 5 Description description page 3 / 3 page 20 CN 121401392 A Abstract The present invention adopts the therapeutic method of warming yang and relieving exterior syndromes to diffuse the lung and promote diuresis, invigorate the spleen and warm the kidney, consolidate defensive qi and relieve edema, and provides a traditional Chinese medicine composition for treating immune-related glomerular diseases, especially membranous nephropathy and minimal change disease. The pharmaceutical composition of the invention is a pure traditional Chinese medicine preparation. Clinical observations as well as in vivo and in vitro basic experiments demonstrate that the prescription can significantly ameliorate edema, massive proteinuria, hypoalbuminemia and other symptoms. Meanwhile, it exerts superior efficacy inalleviating abdominal pain, diarrhea and other manifestations compared with the prior art, and possesses anti-inflammatory activity. No obvious adverse reactions have been observed in clinical application. It achieves particularly remarkable curative effects on patients suffering from membranous nephropathy and minimal change disease differentiated in traditional Chinese medicine as the syndrome of spleen-kidney yang deficiency, latent wind-dampness, and overflowing fluid retention.

Claims

1. A traditional Chinese medicine composition for treating kidney disease, said composition comprising the following traditional Chinese medicines in parts by weight: The herbal composition, consisting of 3-30 parts ephedra, 5-120 parts astragalus, 3-30 parts prepared aconite root, 5-60 parts poria cocos, 5-50 parts cinnamon twig, 5-60 parts stir-fried atractylodes macrocephala, 5-60 parts powdered fangji, 3-50 parts prepared licorice root, 3-50 parts fresh ginger, and 3-50 parts jujube, can significantly improve edema, massive proteinuria, and hypoproteinemia. It also has significant therapeutic effects on symptoms such as abdominal pain and diarrhea, and possesses anti-inflammatory properties.

2. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition comprises the following traditional Chinese medicines in parts by weight: 5-25 parts ephedra, 15-100 parts astragalus, 5-25 parts prepared aconite root, 10-50 parts poria cocos, 10-30 parts cinnamon twig, 10-30 parts stir-fried atractylodes macrocephala, 8-30 parts bupleurum root powder, 8-30 parts prepared licorice root, 5-40 parts fresh ginger, and 5-40 parts jujube.

3. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition comprises the following traditional Chinese medicines in parts by weight: 6-15 parts ephedra, 20-60 parts astragalus, 8-20 parts prepared aconite root, 15-30 parts poria cocos, 10-20 parts cinnamon twig, 15-30 parts stir-fried atractylodes macrocephala, 9-25 parts bupleurum root, 6-15 parts prepared licorice root, 9-15 parts fresh ginger, and 9-15 parts jujube.

4. The traditional Chinese medicine composition as described in claim 1, wherein one or two medicinal and edible herbs may be added to the composition according to the specific symptoms. For example, if sleep is poor, Albizia bark or Ziziphus jujuba seed may be added as adjuvant herbs; if constipation is present, hemp seed or rhubarb may be added; if blood stasis is present, Panax notoginseng powder, Curcuma zedoaria or leech may be added; if sweating is significant, ginseng or white peony root may be added.

5. A traditional Chinese medicine preparation for treating kidney disease, said preparation comprising the following components: 3-30 parts ephedra, 3-30 parts black aconite root slices, 5-120 parts astragalus, 5-60 parts poria cocos, 5-50 parts cinnamon twig, 5-60 parts fangji powder, 5-60 parts stir-fried atractylodes macrocephala, 3-50 parts ginger, 3-50 parts jujube, 3-50 parts prepared licorice root, and excipients required for preparing the corresponding Chinese medicine dosage forms.

6. The traditional Chinese medicine preparation as described in claim 5, wherein the dosage form is one of decoction, tablet, pill, capsule, granule, powder, oral liquid or compound traditional Chinese medicine.

7. A method for preparing a traditional Chinese medicine composition for treating kidney disease, wherein the traditional Chinese medicine composition is obtained by the following preparation method: Preparation method 1 includes the following steps: S01, take the prescribed amount of black aconite slices, soak them in water, adjust the pH, decoct and extract, ultrafilter the extract through an ultrafiltration membrane, concentrate and dry to obtain powder A; S02, take the prescribed amount of ephedra and fangji, pulverize them, soak them in water, adjust the pH, extract them by ultrasonication to obtain the extract, concentrate and dry them to obtain powder B; S03, take the prescribed amount of fresh ginger, wash it, squeeze out the juice, filter it to obtain ginger juice, and set it aside; S04, take the prescribed amount of Astragalus membranaceus, Poria cocos, Cinnamomum cassia, stir-fried Atractylodes macrocephala, roasted Glycyrrhiza uralensis and jujube, add water to decoct, filter, concentrate to clear extract, dry, and obtain powder C; S05, mix the powders A, B and C obtained in steps S01, S02 and S04, add the ginger juice obtained in step S03, mix well, dry, and pulverize to obtain the traditional Chinese medicine composition. Preparation method 2 includes the following steps: S01, take the prescribed amount of black aconite slices, ephedra and fangji powder, crush and mix them, soak them in water, adjust the pH, extract them by ultrasonication to obtain the extract, concentrate and dry them to obtain powder A; S02, take the prescribed amount of fresh ginger, wash it, squeeze out the juice, filter it to obtain ginger juice, and set it aside; S03, take the prescribed amount of Astragalus membranaceus, Poria cocos, Cinnamomum cassia, stir-fried Atractylodes macrocephala, roasted Glycyrrhiza uralensis and jujube, add water and decoct, filter and concentrate to a clear paste, dry to obtain powder B; S04. Mix powder A and powder B from steps S01 and S03 evenly, add the ginger juice obtained in step S02, mix well, dry, and pulverize to obtain the traditional Chinese medicine composition.

8. The use of the traditional Chinese medicine composition of claim 1 or the traditional Chinese medicine preparation of claim 5 in the preparation of a medicament for treating immune nephropathy.

9. The application as described in claim 8, wherein the immune nephropathy includes primary immune nephropathy and secondary immune nephropathy.

10. The application as described in claim 8, wherein the primary immune nephropathy includes membranous nephropathy and minimal change disease. The TCM diagnosis of membranous nephropathy is spleen and kidney yang deficiency with water retention, and the TCM diagnosis of minimal change disease is spleen and kidney yang deficiency with water retention.

Images