Drug concentrated pill for treating chronic kidney disease and preparation method and application thereof

Abstract

This invention relates to a concentrated pill for treating chronic kidney disease, its preparation method, and its application. The raw materials for preparation include the following components: a first extract, a second extract, and pharmaceutically acceptable excipients. The raw materials for preparing the first extract include *Malus kunmingense* (a type of medicinal herb); the raw materials for preparing the second extract include *Epimedium brevicornu*, *Lycium barbarum*, and *Cuscuta chinensis*. This invention improves upon the original dosage form and preparation process of the Kunming-Xian preparation, rationally combining the effective components of each medicinal material to obtain a concentrated pill that effectively treats chronic kidney disease and other diseases, thus better leveraging the clinical efficacy of the Kunming-Xian preparation. Furthermore, through investigation of the extraction and pill-making processes, this invention rationally designs the preparation steps, resulting in a more rational process for the Kunming-Xian concentrated pill, which is more in line with the theoretical basis of traditional Chinese medicine, improves the quality standards of the preparation, has high production efficiency, efficiently utilizes medicinal resources, and has energy-saving, emission-reducing, and environmentally friendly advantages, while also enhancing production safety and meeting the needs of industrial production.

Description

Technical Field

[0001] This application relates to the field of traditional Chinese medicine preparation technology, and in particular to a concentrated pill for treating chronic kidney disease, its preparation method and application. Background Technology

[0002] In recent years, triptolide has become a research hotspot due to its multi-target immunosuppressive effects. Tripterygium wilfordii is the main active ingredient of the traditional Chinese medicine Tripterygium wilfordii, and Tripterygium wilfordii preparations are currently used to treat various diseases, including lupus, tumors, rheumatoid arthritis, and nephrotic syndrome. The main pharmacological effects of triptolide include anti-inflammatory, anti-tumor, and immunosuppressive effects. In addition, studies have shown that triptolide has protective and therapeutic effects on cardiovascular diseases and osteoporosis.

[0003] Currently, the toxic side effects of Tripterygium wilfordii have been a persistent concern in clinical practice. Most documented poisoning incidents are caused by the whole plant, particularly the above-ground parts, which are unsuitable for medicinal use. Regarding the side effects of Tripterygium wilfordii, studies have shown that Tripterygium wilfordii preparations can cause damage to the digestive, reproductive, urinary, and hematopoietic systems. According to previous studies, among 223 patients who used Tripterygium wilfordii, 143 experienced side effects of varying degrees. The most common target organ was the digestive system (35.9%), followed by the reproductive system (22.9%), infection (14.3%), skin and mucous membrane damage (6.7%), and hematopoietic system damage (4.9%).

[0004] Kunxian Capsules are a new compound traditional Chinese medicine for the treatment of rheumatoid arthritis. Based on traditional Chinese medicine theory, it uses *Tripterygium wilfordii*, a renowned herb for treating arthralgia, as its principal ingredient. The formula was developed through years of pharmacodynamic, toxicological, and clinical screening studies of dozens of traditional Chinese medicine compound prescriptions. The principal ingredient, *Tripterygium wilfordii*, is the root or whole plant of *Tripterygium wilfordii*, which has the effects of dispelling wind and dampness, removing blood stasis, and unblocking the meridians. It has long been used in folk medicine to treat rheumatoid arthritis, systemic lupus erythematosus, and chronic nephritis, among other autoimmune diseases, with significant efficacy, but it also has reproductive system toxicity. The combination with *Epimedium brevicornu*, *Lycium barbarum*, and *Cuscuta chinensis*, which tonify the liver and kidneys, not only significantly enhances the wind-dispelling and dampness-removing effects of *Tripterygium wilfordii* but also counteracts its reproductive toxicity, thus achieving synergistic effects and reducing toxicity through the formulation.

[0005] In recent years, multiple clinical studies have shown that Kunxian Capsules are significantly effective in treating kidney disease, possessing the functions of tonifying the kidneys and unblocking the meridians, dispelling wind and dampness. It is primarily used to treat rheumatoid arthritis with a syndrome of wind-dampness obstruction and kidney deficiency. It can significantly improve proteinuria and serum immunoglobulin levels in kidney disease patients, with no major adverse events. Compared to immunosuppressants such as glucocorticoids, it has better efficacy and fewer side effects. The production process of Kunxian Capsules involves extracting *Trichosanthes kirilowii*, *Epimedium brevicornu*, *Lycium barbarum*, and *Cuscuta chinensis* with pure water. The extract is then separated and purified using macroporous adsorption resin. The eluent is collected, concentrated under reduced pressure, dried to a suitable amount, and then dried using a spray dryer to obtain a dry powder of the four herbs. This powder is then mixed with excipients, granulated, and encapsulated to obtain the product. The main limiting factor in its production is the use of macroporous resin, and the following problems exist:

[0006] 1. The elution time for each of the four medicinal ingredients in the production process is 13-18 hours, which is the most time-consuming stage in the entire production process. 2. Resin elution requires a large amount of 70% ethanol and must be carried out in an explosion-proof workshop, resulting in high production costs and safety risks. 3. The original process of Kunxian Capsules used macroporous resin to purify and enrich the aqueous extracts of Lycium barbarum and Cuscuta chinensis. This process discharges a large amount of waste liquid containing polysaccharides and other macromolecular substances, causing the COD of the production wastewater to exceed the standard, reaching 60,000 mg / L. The wastewater could not be discharged in time and required membrane separation and concentration, and multiple batches had to be collected before discharge. Furthermore, the waste liquid is prone to fermentation and deterioration, resulting in poor air quality in the factory. 4. The original process used macroporous adsorption resin to separate and purify Epimedium, Lycium barbarum, and Cuscuta chinensis, resulting in the loss of water-soluble components. Therefore, the indicative components betaine and hyperoside of the two medicinal ingredients listed in the 2020 edition of the Chinese Pharmacopoeia could not be detected in the formulation.

[0007] In summary, there is an urgent need to improve the original dosage form and process of Kunxian capsules in order to better meet production needs and to better exert the clinical efficacy of Kunxian preparations. Summary of the Invention

[0008] Therefore, it is necessary to provide a concentrated pill for treating chronic kidney disease, its preparation method and application. By changing the dosage form of Kunxian preparation from capsules to pills and improving the preparation process, the production efficiency is improved, and the preparation is more environmentally friendly, energy-saving and safe while ensuring the clinical efficacy of Kunxian preparation.

[0009] The technical solution is as follows:

[0010] In a first aspect, the present invention provides a concentrated pill for treating chronic kidney disease, the raw materials for which are prepared include the following components: a first extract, a second extract, and pharmaceutically acceptable excipients;

[0011] The raw materials for preparing the first extract include *Malus kunmingensis*;

[0012] The raw materials for preparing the second extract include epimedium, wolfberry, and dodder seed;

[0013] The first extract, the second extract, a mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to form a concentrated drug pill;

[0014] The second extract includes: dodder seed powder, and an extract prepared by extracting one or more of epimedium, wolfberry and dodder seed individually or in combination.

[0015] In one embodiment, the concentrated pill for treating chronic kidney disease comprises the following parts by weight of active pharmaceutical ingredient:

[0016] 1-3 parts of *Malus kunmingensis*, 1-3 parts of *Epimedium brevicornu*, 0.5-1.5 parts of *Lycium barbarum*, and 0.5-1.5 parts of *Cuscuta chinensis*.

[0017] In one embodiment, the concentrated pill for treating chronic kidney disease comprises the following parts by weight of active pharmaceutical ingredient:

[0018] Two portions of *Malus kunmingensis*, two portions of *Epimedium brevicornu*, one portion of *Lycium barbarum*, and one portion of *Cuscuta chinensis*.

[0019] In one embodiment, the first extract comprises dried powder of *Malus kunmingensis*; the second extract comprises: an extract prepared by combining the extraction of *Epimedium brevicornu*, *Lycium barbarum* and *Cuscuta chinensis*, or an extract prepared by combining the extraction of *Epimedium brevicornu*, *Lycium barbarum* and *Cuscuta chinensis* separately, or an extract prepared by combining the extraction of *Epimedium brevicornu* and *Lycium barbarum* separately and *Cuscuta chinensis* powder, or an extract prepared by combining the extraction of *Epimedium brevicornu* and *Lycium barbarum* separately and *Cuscuta chinensis* powder.

[0020] In one embodiment, the second extract comprises: a mixture of powdered dodder seed and extracts of the other two herbs, epimedium and wolfberry, prepared individually or in combination.

[0021] In one embodiment, the pharmaceutically acceptable excipient includes a mixture of starch and microcrystalline cellulose.

[0022] In one embodiment, the pharmaceutically acceptable excipients contain starch and microcrystalline cellulose in a mass ratio of (1-3):1.

[0023] In one embodiment, the concentrated pill for treating chronic kidney disease is prepared from the active ingredients in the raw materials.

[0024] Wherein, the Kunming crabapple is replaced by one or more of the diterpenoids, triterpenoids and alkaloids contained therein;

[0025] The epimedium is replaced by one or more of the following: epimedium glycoside, epimedium glycoside I, epimedium glycoside II, and epimedium glycoside A contained therein;

[0026] The dodder seed and wolfberry are replaced by one or more of the flavonoids and polysaccharides contained therein.

[0027] A second aspect of the present invention provides a method for preparing a concentrated pill for treating chronic kidney disease as described above, comprising the following steps:

[0028] Provides dried crabapple extract powder from Kunming;

[0029] The second solvent was used to extract Epimedium and Lycium barbarum separately or in combination, and the resulting extracts were concentrated to prepare an extract.

[0030] Dodder seeds are crushed to prepare dodder powder;

[0031] The dodder seed powder, dried crabapple extract powder, extract, mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to form concentrated drug pills.

[0032] The drug concentrate pill is Kunxian concentrate pill, and the second solvent is water.

[0033] In one embodiment, the step of using a second extraction solvent to perform a combined extraction of Epimedium and Lycium barbarum includes at least one of the following conditions:

[0034] (1) The extraction method was heating and reflux extraction;

[0035] (2) The extraction time is 1-3 hours;

[0036] (3) The volume ratio of the second solvent to the total mass of Epimedium and Lycium barbarum is (10-15) L: 1 kg;

[0037] (4) The mass ratio of the dodder powder to the pharmaceutically acceptable excipient is (1-3):1.

[0038] In one embodiment, the method for preparing the concentrated drug pill for treating chronic kidney disease includes the following steps:

[0039] Provides dried crabapple extract powder from Kunming;

[0040] The second solvent was used to extract Epimedium, Lycium barbarum and Cuscuta chinensis individually or in combination, and the resulting extracts were concentrated to prepare an extract.

[0041] The Kunming mountain crabapple dried extract powder, extract, mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to prepare concentrated drug pills;

[0042] The drug concentrate pill is Kunxian concentrate pill, and the second solvent is water.

[0043] In one embodiment, the preparation method of the dried extract powder of *Malus kunmingensis* includes the following steps:

[0044] The first solvent was used to extract the *Malus kunmingensis*, and the resulting extract was separated, purified, and dried to prepare *Malus kunmingensis* dry extract powder.

[0045] The first solvent is water.

[0046] In one embodiment, the preparation method of the dried extract powder of *Malus kunmingensis* includes the following steps:

[0047] Weigh out the crabapple from Kunming Mountain, add water and heat under reflux to extract three times; load the obtained extract into a macroporous resin for adsorption, and after adsorption, elute with 70% ethanol solution; concentrate the obtained eluent under reduced pressure and dry.

[0048] A third aspect of the present invention provides the use of the drug concentrate pills for treating chronic kidney disease as described above in the preparation of drugs for treating chronic kidney disease, rheumatoid arthritis, systemic lupus erythematosus, Crohn's disease, or leprosy reactions.

[0049] In one embodiment, the chronic kidney disease includes IgA nephropathy.

[0050] Compared with the prior art, this application has the following beneficial effects:

[0051] This application improves the original dosage form of Kunxian preparation by obtaining a concentrated pill that can effectively treat diseases such as chronic kidney disease based on the properties of the effective components in each medicinal material. The raw materials for its preparation include a first extract and a second extract. The first extract and the second extract are mixed and pharmaceutically acceptable excipients are added. Through reasonable compatibility, the clinical efficacy of Kunxian preparation can be better exerted.

[0052] Meanwhile, this application improves the Kunxian capsule process through extraction and pill-making process investigation, and proposes a preparation method for Kunxian concentrated pills. This method rationally designs the preparation steps based on the properties of the effective components in each medicinal material, retains the water-soluble components of Epimedium and Lycium barbarum, and makes the process more reasonable and more in line with the theoretical basis of traditional Chinese medicine. It improves the quality standards of the preparation, has high production efficiency, makes efficient use of Chinese medicinal resources, and has the advantages of energy saving, consumption reduction and environmental protection. It also has higher production safety and meets the needs of industrial production. Attached Figure Description

[0053] To more clearly illustrate the technical solutions in the specific embodiments of this application or the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0054] Figure 1 This is a stacked diagram showing the results of triptolide content determination in one embodiment.

[0055] Figure 2 This is a standard curve and determination coefficient diagram of triptolide in one embodiment.

[0056] Figure 3 This is a chromatogram showing the results of icariin content determination in one embodiment.

[0057] Figure 4 This is a standard curve and determination coefficient diagram of icariin in one embodiment.

[0058] Figure 5 This is a chromatogram for the determination of triptolide content in a mixture of four medicinal materials extracted with water in one embodiment.

[0059] Figure 6 This is a diagram showing the standard curve and correction coefficient of triptolide extracted from a mixture of four medicinal herbs in one embodiment.

[0060] Figure 7 This is a chromatogram for the determination of triptolide content in water extracted from *Malus kunmingensis* in one embodiment.

[0061] Figure 8 This is a diagram showing the standard curve and correction coefficient of triptolide extracted from *Malus kunmingensis* alone in one embodiment.

[0062] Figure 9 The image shows a chromatogram for the determination of triptolide content extracted from *Malus kunmingensis* using 50% ethanol in one embodiment.

[0063] Figure 10 This is a standard curve and correction coefficient diagram for the extraction of triptolide from 50% ethanol alone in one embodiment of *Malus kunmingensis*.

[0064] Figure 11 The image shows a chromatogram for the determination of triptolide content in the 50% ethanol extract of four medicinal herbs, Kunshan and Xianshan, in one embodiment.

[0065] Figure 12 The figure shows the standard curve and correction coefficient diagram for the extraction of triptolide from four medicinal herbs (Kunshan, Xian, and Dioscorea) using 50% ethanol in one embodiment.

[0066] Figure 13 This is a chromatogram for the determination of icariin content extracted from Epimedium alone in one embodiment.

[0067] Figure 14 This is a standard curve and correction coefficient diagram for the extraction of icariin from epimedium alone in one embodiment.

[0068] Figure 15 This is a standard curve and determination coefficient diagram of the pilot-scale product icariin in one embodiment.

[0069] Figure 16 This is a chromatographic overlay of a pilot-scale product for the determination of icariin content in one embodiment.

[0070] Figure 17 This is a standard curve and determination coefficient diagram of the pilot-scale product triptolide in one embodiment.

[0071] Figure 18 This is a chromatographic overlay diagram for the determination of triptolide content in a pilot-scale product in one embodiment.

[0072] Figure 19 This is a flowchart of the preparation process for Kunxian capsules and Kunxian concentrated pills.

[0073] Figure 20 The image shows a section of rat kidney pathological tissue (×100) from each group in one example. Detailed Implementation

[0074] The present application will be further described in detail below with reference to specific embodiments. The present application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of this application.

[0075] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0076] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0077] In this article, "one or more" refers to any one, two or more of the listed items.

[0078] In this document, the optional range of "and / or", "or / and", and "and / or" includes any one of two or more of the related listed items, as well as any and all combinations of the related listed items, including any two related listed items, any more related listed items, or a combination of all the related listed items.

[0079] In this document, terms such as “further,” “even further,” and “especially” are used to describe the purpose and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0080] In this document, terms such as "first aspect," "second aspect," "third aspect," and "fourth aspect" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly specifying the importance or quantity of the indicated technical features. Furthermore, terms such as "first," "second," "third," and "fourth" serve only as a non-exhaustive enumeration and should be understood as not constituting a closed limitation on quantity. Therefore, a feature specified as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified. In the description of this application, "several" means at least one, such as one, two, etc., unless otherwise explicitly specified.

[0081] In this application, numerical ranges are referred to as continuous unless otherwise specified, and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0082] Unless otherwise specified, the percentage content mentioned in this application refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures.

[0083] Unless otherwise specified, all percentage concentrations mentioned in this application refer to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of that component.

[0084] Unless otherwise specified, the temperature parameters in this application allow for both isothermal processing and processing within a certain temperature range. The isothermal processing allows temperature fluctuations within the precision range of instrument control, such as ±5℃, ±2℃, ±1℃, ±0.5℃, ±0.4℃, ±0.3℃, ±0.2℃, and ±0.1℃. In this application, room temperature or ambient temperature refers to the absence of temperature control operations, generally meaning 4℃ to 35℃, and preferably 20±5℃.

[0085] In a first aspect, the present invention provides a concentrated pill for treating chronic kidney disease, the raw materials for which are prepared include the following components: a first extract, a second extract, and pharmaceutically acceptable excipients;

[0086] The raw materials for preparing the first extract include *Malus kunmingensis*;

[0087] The raw materials for preparing the second extract include epimedium, wolfberry, and dodder seed;

[0088] The first extract, the second extract, a mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to form concentrated drug pills;

[0089] The second extract includes: dodder seed powder, and an extract prepared by extracting one or more of epimedium, wolfberry and dodder seed individually or in combination.

[0090] In some of these examples, the drug concentrate for treating chronic kidney disease comprises the following parts by weight of active pharmaceutical ingredient:

[0091] 1-3 parts of *Malus kunmingensis*, 1-3 parts of *Epimedium brevicornu*, 0.5-1.5 parts of *Lycium barbarum*, and 0.5-1.5 parts of *Cuscuta chinensis*.

[0092] In some of these examples, the drug concentrate for treating chronic kidney disease comprises the following parts by weight of active pharmaceutical ingredient:

[0093] Two portions of *Malus kunmingensis*, two portions of *Epimedium brevicornu*, one portion of *Lycium barbarum*, and one portion of *Cuscuta chinensis*.

[0094] In some examples, the first extract includes dried powder of *Malus kunmingensis*; the second extract includes: an extract prepared by combining the extraction of *Epimedium brevicornu*, *Lycium barbarum* and *Cuscuta chinensis*, or an extract prepared by combining the extraction of *Epimedium brevicornu*, *Lycium barbarum* and *Cuscuta chinensis* separately, or an extract prepared by combining the extraction of *Epimedium brevicornu* and *Lycium barbarum* separately and *Cuscuta chinensis* powder, or an extract prepared by combining the extraction of *Epimedium brevicornu* and *Lycium barbarum* separately and *Cuscuta chinensis* powder.

[0095] In some examples, the second extract comprises: a mixture of powdered dodder seed and extracts of the other two herbs, epimedium and wolfberry, extracted individually or in combination.

[0096] In some of these examples, the pharmaceutically acceptable excipients include a mixture of starch and microcrystalline cellulose.

[0097] In some of these examples, the pharmaceutically acceptable excipients contain starch and microcrystalline cellulose in a mass ratio of (1-3):1. It is understood that the mass ratio of starch to microcrystalline cellulose in the pharmaceutically acceptable excipients includes, but is not limited to, 1:1, 2:1, and 3:1.

[0098] In some examples, the concentrated pills for treating chronic kidney disease are prepared from the active ingredients in the raw materials used in the preparation;

[0099] Wherein, the Kunming crabapple is replaced by one or more of the diterpenoids, triterpenoids and alkaloids contained therein;

[0100] The epimedium is replaced by one or more of the following: epimedium glycoside, epimedium glycoside I, epimedium glycoside II, and epimedium glycoside A contained therein;

[0101] The dodder seed and wolfberry are replaced by one or more of the flavonoids and polysaccharides contained therein.

[0102] A second aspect of the present invention provides a method for preparing a concentrated pill for treating chronic kidney disease as described above, comprising the following steps:

[0103] Provides dried crabapple extract powder from Kunming;

[0104] The second solvent was used to extract Epimedium and Lycium barbarum separately or in combination, and the resulting extracts were concentrated to prepare an extract.

[0105] Dodder seeds are crushed to prepare dodder powder;

[0106] The dodder seed powder, dried crabapple extract powder, extract, mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to form concentrated drug pills.

[0107] The drug concentrate pill is Kunxian concentrate pill, and the second solvent is water.

[0108] The original preparation process of Kunxian Capsules involved extracting Kunming Mountain Crabapple, Epimedium, Lycium barbarum, and Cuscuta chinensis with pure water, followed by separation and purification using macroporous adsorption resin, and then spray drying to obtain a dry powder. This powder was then mixed, granulated, and encapsulated to obtain the product. Based on this, the technical personnel of this application proposed a new preparation process for pills by changing the preparation process and dosage form of Kunxian Capsules: Kunming Mountain Crabapple is extracted with solvent, then separated, purified, and dried to obtain a dry powder. Epimedium and Lycium barbarum are extracted with solvent and concentrated into an extract. Cuscuta chinensis is powdered, and the powder and extract are mixed, dried, and then mixed with the Kunming Mountain Crabapple dry powder and excipients to form pills. This process changes the original capsule formulation to a pill formulation, retaining the water-soluble components of Epimedium and Lycium barbarum. The powdering of Cuscuta chinensis reduces the use of excipients, while retaining all components of Cuscuta chinensis. The process is more rational in terms of the prescription ingredients, improving the quality standards of the preparation. Meanwhile, the technicians of this application conducted pharmacodynamic and immunological mechanism studies on the drug composition in IgA nephropathy rats using a pill dosage form. An IgA nephropathy model was established using a combined drug administration method of "bovine serum albumin + carbon tetrachloride + lipopolysaccharide". The pharmacological effects and immunological mechanisms were evaluated using proteinuria, creatinine, blood urea nitrogen, cytokines, immunoglobulin levels, and histopathological examination as indicators. The results showed that the Kunxian concentrated pills prepared by the new process had a significant therapeutic effect on IgA nephropathy rats, reducing proteinuria levels and renal function indicators, alleviating kidney tissue damage, and the effect was better than the original Kunxian capsules under the same raw drug dosage.

[0109] In summary, this application improves the Kunxian capsule process through extraction and pill-making process investigation, proposing a method for preparing Kunxian concentrated pills. This method rationally designs the preparation steps based on the properties of the effective components in each medicinal material. The process involves separately extracting and purifying *Trichosanthes kirilowii* to prepare a dry extract powder, then extracting and concentrating *Epimedium brevicornu* and *Lycium barbarum* together to form an extract, pulverizing *Cuscuta chinensis* and mixing it with the extract, drying, and pulverizing to obtain a mixed dry extract powder. The *Trichosanthes kirilowii* dry extract powder is then mixed with excipients and further mixed with the dry extract powders of the remaining medicinal ingredients to form pills. This process is more rational, more in line with the theoretical basis of traditional Chinese medicine, and can enhance the combination of medicinal ingredients to reduce toxicity and increase efficacy, improve the quality standards of the preparation, and has high production efficiency. It also has advantages such as efficient utilization of medicinal resources, energy saving, consumption reduction, and environmental protection, meeting the needs of industrial production. Furthermore, pharmacological studies have shown that the Kunxian concentrated pills prepared by this method have significant therapeutic effects on IgA nephropathy rats, reducing proteinuria levels and renal function indicators, alleviating kidney tissue damage, and with the same amount of raw drug, the effect is better than the original Kunxian capsules, showing broad application prospects in clinical practice.

[0110] In some of these examples, the conditions for the combined extraction of Epimedium and Lycium barbarum using a second extraction solvent include: the extraction method being reflux extraction.

[0111] In some examples, the conditions for the combined extraction of Epimedium and Lycium barbarum using a second extraction solvent include an extraction time of 1 to 3 hours. Understandably, the extraction time includes, but is not limited to, 1 hour, 2 hours, and 3 hours.

[0112] In some examples, the conditions for the combined extraction of Epimedium and Lycium barbarum using a second extraction solvent include: the volume ratio of the second extraction solvent to the total mass of Epimedium and Lycium barbarum is (10-15) L: 1 kg. Understandably, the volume ratio of the second extraction solvent to the total mass of Epimedium and Lycium barbarum includes, but is not limited to, 10 L: 1 kg, 11 L: 1 kg, 12 L: 1 kg, 13 L: 1 kg, 14 L: 1 kg, and 15 L: 1 kg.

[0113] In some of these examples, the mass ratio of the dodder seed powder to the pharmaceutically acceptable excipient is (1-3):1. It is understood that the mass ratio of the dodder seed powder to the pharmaceutically acceptable excipient includes, but is not limited to, 1:1, 2:1, and 3:1.

[0114] In some of these examples, the method for preparing the concentrated pills for treating chronic kidney disease includes the following steps:

[0115] Provides dried crabapple extract powder from Kunming;

[0116] The second solvent was used to extract Epimedium, Lycium barbarum and Cuscuta chinensis individually or in combination, and the resulting extracts were concentrated to prepare an extract.

[0117] The Kunming mountain crabapple dried extract powder, extract, mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to prepare concentrated drug pills;

[0118] The drug concentrate pill is Kunxian concentrate pill, and the second solvent is water.

[0119] In some examples, the preparation method of the dried extract powder of *Malus kunmingensis* includes the following steps:

[0120] The first solvent was used to extract the *Malus kunmingensis*, and the resulting extract was separated, purified, and dried to prepare *Malus kunmingensis* dry extract powder.

[0121] In some of these examples, the first solvent is water.

[0122] In some examples, the preparation method of the dried extract powder of *Malus kunmingensis* includes the following steps:

[0123] Weigh out the crabapple from Kunming Mountain, add water and heat under reflux to extract three times; load the obtained extract into a macroporous resin for adsorption, and after adsorption, elute with 70% ethanol solution; concentrate the obtained eluent under reduced pressure and dry.

[0124] A third aspect of the present invention provides the use of the drug concentrate pills for treating chronic kidney disease as described above in the preparation of drugs for treating chronic kidney disease, rheumatoid arthritis, systemic lupus erythematosus, Crohn's disease, or leprosy reactions.

[0125] In some of these examples, the chronic kidney disease includes IgA nephropathy.

[0126] Chronic kidney disease (CKD) is one of the major diseases threatening the health of the Chinese population. A recent epidemiological survey in my country showed a CKD incidence rate as high as 10.8%. Although hypertensive nephropathy and diabetic nephropathy have significantly increased the incidence of CKD, primary glomerulonephritis remains the leading cause of CKD in my country. How to actively treat primary glomerular diseases and slow the progression of CKD is a serious clinical challenge.

[0127] IgA nephropathy (IgAN) is the most common primary glomerulonephritis in clinical practice, especially prevalent in young and middle-aged patients. It is characterized by predominantly IgA or IgA deposition in the glomerular mesangial area, with or without the deposition of other immunoglobulins, and is an immune complex-mediated disease. The clinical manifestations of IgA nephropathy vary greatly, ranging from asymptomatic hematuria with or without proteinuria to nephrotic syndrome, and even some patients present with rapidly progressive glomerulonephritis. However, more than one-third of IgA nephropathy patients will eventually progress to end-stage renal disease within 10 years of onset. Current research suggests that the risk factors for progression to end-stage renal disease include uncontrolled hypertension, proteinuria exceeding 1 g / day, and elevated serum creatinine levels. Clinical treatment for IgA nephropathy mainly includes glucocorticoids and immunosuppressants. Although glucocorticoids and immunosuppressants can effectively reduce proteinuria and slow the progression of kidney disease, studies have also found that their use significantly increases the incidence of adverse reactions, especially serious infections. In clinical practice, individualized treatment is crucial for different patients. Treatment plans should be developed based on a thorough assessment of patient benefits and adverse event risks, and interventions focused on protecting the kidneys are essential for patient benefit. Therefore, it is urgent to find suitable alternative drugs that reduce the adverse reactions caused by long-term hormone therapy while actively treating kidney disease, and that offer relatively fewer adverse reactions and higher cost-effectiveness. The Kunxian concentrated pills provided in this application retain the water-soluble components of Epimedium, Lycium barbarum, and Cuscuta chinensis. Studies have shown that the water-soluble components of Lycium barbarum and Cuscuta chinensis have immunomodulatory and toxicity-mitigating effects, which help enhance the liver- and kidney-tonifying, immune-regulating, and toxicity-reducing effects of the adjuvant herbs in the formula, thus better leveraging the compatibility of the adjuvant herbs. The production process is also more aligned with the theoretical basis of traditional Chinese medicine.

[0128] Specifically, this application uses a combined drug therapy of bovine serum albumin, carbon tetrachloride, and lipopolysaccharide to establish an IgA nephropathy model. This model closely resembles the symptoms of proteinuria in clinical IgA nephropathy patients and is a commonly used disease model in the field of nephrology research. Results show that all drug-treated groups can reduce 24-hour urinary protein, thymus index, urinary white blood cell count, urinary bilirubin, urinary protein content, alanine aminotransferase, aspartate aminotransferase, liver index, kidney index, blood urea nitrogen, creatinine, interleukin-1β, interleukin-6, and tumor necrosis factor-α to varying degrees (P<0.05), and improve the proliferation of glomerular mesangial cells and increased matrix.

[0129] The following detailed description is provided in conjunction with specific embodiments. Unless otherwise specified, all raw materials used in the following embodiments are commercially available; all instruments used are commercially available; and all processes involved are conventionally selected by those skilled in the art unless otherwise specified.

[0130] Example 1

[0131] This case study examines the extraction and pelleting processes of Kunxian concentrated pills, as detailed below:

[0132] 1. Investigation of extraction process

[0133] 1.1 Determination of the content of index components in medicinal materials

[0134] 1.1.1 Determination of the medicinal content of *Malus kunmingensis*

[0135] (1) Experimental instruments and reagents

[0136] Experimental instruments: Agilent 1260 high performance liquid chromatograph, HWS-26 electric thermostatic water bath;

[0137] Experimental reagents: Kunming mountain crabapple medicinal material, 95% ethanol, 1,2-dichloroethane, methanol, silica gel, neutral alumina, triptolide.

[0138] (2) Experimental methods

[0139] Extraction method: Weigh three portions of Kunming mountain crabapple powder (passed through a 65-mesh sieve), each approximately 1.5g, accurately. Place in a Soxhlet extractor, add an appropriate amount of 95% ethanol, reflux for 8 hours, transfer the ethanol solution to an evaporating dish, evaporate to dryness, dissolve in a small amount of methanol, mix with 2g of silica gel (for column chromatography, 200-300 mesh) - neutral alumina (for column chromatography, 200-300 mesh) (1:1), evaporate to dryness, and dry-load onto a silica gel (for column chromatography, 200-300 mesh) - neutral alumina (for column chromatography, 200-300 mesh) (1:1, 2g) mixed column (20mm inner diameter). Elute with 50 mL of 1,2-dichloroethane, discard the eluent, and continue eluting with 50 mL of 1,2-dichloroethane in 1% ethanol (95% ethanol). Collect the eluent, evaporate to dryness, dissolve the residue in methanol and transfer it to a 5 mL volumetric flask. Dilute to the mark with methanol and shake well to obtain the final product.

[0140] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (40:60) was used as the mobile phase; and the detection wavelength was 225 nm. An appropriate amount of triptolide reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 5.3 μg per mL. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed.

[0141] (3) Experimental Results

[0142] The results are shown in Table 1. Figure 1 and Figure 2As shown in the table, samples 1 and 2 refer to two parallel determination experiments of the Kunming Mountain crabapple medicinal material used in the experiment to ensure the accuracy of the content determination (the sample numbers in other tables also refer to parallel determination experiments). The results show that the content of triptolide in the Kunming Mountain crabapple medicinal material is 0.0153‰.

[0143] Table 1. Results of Tripterygium wilfordii content detection in Kunming crabapple medicinal material.

[0144]

[0145]

[0146] 1.1.2 Determination of Epimedium content in medicinal materials

[0147] (1) Experimental instruments and reagents

[0148] Experimental instruments and reagents: Agilent 1200 high performance liquid chromatograph, SB-5200DT ultrasonic cleaner, Sartorius SQP balance, HG53 moisture analyzer;

[0149] Experimental reagents: Epimedium medicinal material, epimedin, 95% ethanol.

[0150] (2) Experimental methods

[0151] Extraction method: Weigh two portions of Epimedium powder (passed through a 65-mesh sieve), each approximately 0.2g, accurately place them in a stoppered conical flask, accurately add 20mL of 50% ethanol solution, and accurately weigh again. Sonicate for 1 hour, weigh again, and use the 50% ethanol solution to compensate for any weight loss. Shake well, filter, and collect the filtrate to obtain the final product.

[0152] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; acetonitrile-water (30:70) was used as the mobile phase; and the detection wavelength was 270 nm. An appropriate amount of icariin reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 0.1 mg / mL. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed. The theoretical plate number, calculated based on the icariin peak, should not be less than 1500.

[0153] (3) Experimental Results

[0154] The results are shown in Table 2. Figure 3 and Figure 4 As shown, the content of epimedin in the herbal medicine Epimedium is 0.7481%.

[0155] Table 2. Results of Epimedium content determination in Epimedium medicinal material.

[0156]

[0157] 1.2 Determining the extraction process route of medicinal ingredients through extraction process investigation

[0158] 1.2.1 Research on Mixed Pure Water Extraction Process

[0159] (1) Experiment on the mixed extraction of 4 medicinal materials

[0160] Experimental medicinal materials: Kunming crabapple, epimedium, wolfberry, dodder seed;

[0161] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath;

[0162] Experimental Method: Weigh 0.5 times the prescribed amount of medicinal materials: 50g of *Tripterygium wilfordii*, 50g of *Epimedium brevicornu*, 25g of *Lycium barbarum*, and 25g of *Cuscuta chinensis*. Place them in a 5000mL round-bottom flask, add 1800mL of drinking water, soak for 30 minutes, reflux for 1 hour, and pour off the extract. Add 1500mL of drinking water to the residue, reflux for 1 hour, and pour off the extract. Combine the two extracts and measure their volume and weight. A portion of the extract is used for the determination of the dry extract weight and triptolide content. (The second step is a repetition of the first step.)

[0163] The experimental results are shown in Table 3. The results show that after two extractions of the four medicinal materials, the dry extract rate was 22.04%; after a third extraction of the residue, the dry extract rate was 2.08%.

[0164] Table 3 Results of Dry Extract Yield Determination

[0165]

[0166] Tripterygium wilfordii content determination

[0167] Experimental instruments: Agilent 1260, EYEL4 rotary evaporator, SB-5200DT ultrasonic cleaner, HWS-26 electric thermostatic water bath, Sartorius BS110S electronic balance;

[0168] Experimental reagents: 95% ethanol, 1,2-dichloroethane, methanol, silica gel, neutral alumina, triptolide;

[0169] Experimental methods:

[0170] Pretreatment method: Transfer two portions of extract to a 150mL round-bottom flask, concentrate under reduced pressure in a rotary evaporator until completely dry, add 50mL methanol solution, sonicate to dissolve, transfer to a 50mL centrifuge tube, centrifuge at 10000r / min for 10min, transfer the supernatant to an evaporating dish, dry in an 80℃ water bath to an appropriate amount, and determine the triptolide content in the *Tripterygium wilfordii* medicinal material according to the method for determining triptolide content in *Tripterygium wilfordii*.

[0171] Accurately add 50 mL of methanol to a round-bottom flask, weigh it, sonicate (300 W, 40 kHz) for 15 minutes, cool, stopper tightly, and let stand overnight. Accurately pipette 25 mL of the supernatant into an evaporating dish, place it in a 60°C water bath and evaporate to about 3 mL. Add 2.5 g of silica gel (for column chromatography, 200–300 mesh) – neutral alumina (for chromatography, 200–300 mesh) (1:1), mix well, evaporate the methanol in a 60°C water bath while grinding finely, and evaporate the methanol completely. Dry-press the mixture onto a 2 cm inner diameter column containing 2.5 g of a silica gel (for column chromatography, 200–300 mesh) – neutral alumina (for chromatography, 200–300 mesh) (1:1) mixture. Elute with 70 mL of 1,2-dichloroethane, discard the eluent, and then elute with 70 mL of 1,2-dichloroethane solution containing 1.5% ethanol. Collect the eluent, recover the solvent under reduced pressure at 60°C until dry, dissolve in methanol and transfer to a 5 mL volumetric flask, add methanol to the mark, shake well, and filter to obtain the final product.

[0172] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (5% isopropanol) (18:82) was used as the mobile phase; the detection wavelength was 220 nm. An appropriate amount of triptolide reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 7.0 μg per ml. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed.

[0173] The experimental results are shown in Table 4. Figure 5 and Figure 6 As shown, the results indicate that after two extractions of the four medicinal materials, the extraction rate of triptolide was 5.811 μg / g, and after a third extraction of the residue, the extraction rate of triptolide was 1.3578 μg / g.

[0174] Table 4. Results of determination of triptolide content in extract.

[0175]

[0176] (2) Extraction of *Malus kunmingensis* alone, and extraction of *Epimedium brevicornu*, *Lycium barbarum*, and *Cuscuta chinensis* in combination.

[0177] Extraction experiment:

[0178] Experimental medicinal materials: Kunming crabapple, epimedium, wolfberry, dodder seed;

[0179] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath;

[0180] Experimental Method: Weigh out one prescription amount of the following herbs: 100g of *Trichosanthes kirilowii*, 100g of *Epimedium brevicornu*, 50g of *Lycium barbarum*, and 50g of *Cuscuta chinensis*. Extract *Trichosanthes kirilowii* separately, and extract the other three herbs together. Place *Trichosanthes kirilowii* in a 2000mL round-bottom flask, add 1200mL of drinking water, soak for 30 minutes, reflux for 1 hour, and discard the extract. Add 1000mL of drinking water to the residue, reflux for 1 hour, discard the extract, combine the two extracts, and measure the volume and weight. A portion of the extract is used for the determination of the dry extract weight and triptolide content. (The second step is a repetition of the first step.)

[0181] Place Epimedium, Lycium barbarum, and Cuscuta chinensis in a 5000mL round-bottom flask, add 2400mL of drinking water, soak for 30 minutes, reflux for 1 hour, and discard the extract. Add 2000mL of drinking water to the residue, reflux for 1 hour, and discard the extract. Combine the two extracts and measure their volume and weight. A portion of the extract is used for determining the weight of the dry extract. Divide the remaining extract into two portions and concentrate them separately under normal and reduced pressure to prepare the dry extract for later use. (The last sentence is a repetition of the previous one and can be omitted.)

[0182] The experimental results are shown in Table 5. The results indicate that extracting *Trichosanthes kirilowii* alone twice yielded a dry extract yield of 10.42%, while extracting the residue a third time yielded a dry extract yield of 1.08%. Extracting the remaining three herbs—Epimedium, Lycium barbarum, and Cuscuta chinensis—combined twice yielded a dry extract yield of 26.51%, while extracting the residue a third time yielded a dry extract yield of 2.85%.

[0183] Table 5 Results of Dry Extract Yield Measurement

[0184]

[0185] Determination of Tripterygium wilfordii content in extract

[0186] Experimental instruments: Agilent 1260, EYEL4 rotary evaporator, SB-5200DT ultrasonic cleaner, HWS-26 electric thermostatic water bath, Sartorius BS110S electronic balance;

[0187] Experimental reagents: 1,2-dichloroethane, methanol, silica gel, neutral alumina, triptolide;

[0188] Experimental methods:

[0189] Pretreatment method: Transfer the measured 4 portions of extract to 150mL round-bottom flasks, concentrate under reduced pressure in a rotary evaporator until completely dry, add 50mL methanol solution, sonicate to dissolve, transfer to 50mL centrifuge tubes, centrifuge at 10000r / min for 10min, transfer the supernatant to an evaporating dish, dry in an 80℃ water bath to an appropriate amount, and determine the triptolide content in the *Tripterygium wilfordii* medicinal material according to the method for determining triptolide content in *Tripterygium wilfordii*.

[0190] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (40:60) was used as the mobile phase; and the detection wavelength was 225 nm. An appropriate amount of triptolide reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 5.3 μg per mL. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed.

[0191] The experimental results are shown in Table 6. Figure 7 and Figure 8 As shown, the extraction rate of triptolide from *Malus kunmingensis* alone was 6.5195 μg / g after two extractions, and 1.0114 μg / g after a third extraction from the residue. This indicates that the extraction rate of triptolide from *Malus kunmingensis* alone was higher than that from the mixed extraction method of the four herbs.

[0192] Table 6. Results of determination of triptolide content in extract.

[0193]

[0194] 1.2.2 Study on 50% Ethanol Extraction Process

[0195] Research on 50% Ethanol Extraction Process of Malus baccata in Kunming

[0196] Experimental medicinal material: Kunming crabapple;

[0197] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath, EYEL4 rotary evaporator;

[0198] Experimental Method: Weigh 100g of *Malus kunmingensis* (one prescription amount) into a round-bottom flask, add 12 times its volume of 50% ethanol, soak for 30 minutes, reflux for 1 hour, pour off the extract, add 10 times its volume of 50% ethanol to the residue, reflux for 1 hour, pour off the extract, add 10 times its volume of 50% ethanol to the residue, reflux for 1 hour, pour off the extract, combine the three extracts, and measure their volume and weight. A portion of the extract is used for the determination of the weight and content of the dry extract. The remaining extract is concentrated under reduced pressure to prepare the dry extract for later use.

[0199] The experimental results are shown in Table 7. The results show that the yield of dry extract of *Malus kunmingensis* extracted with 50% ethanol alone is 12.68%.

[0200] Table 7. Results of the determination of 50% ethanol extract of *Malus kunmingensis* (Kunming crabapple) dry extract.

[0201]

[0202] Tripterygium wilfordii content determination

[0203] Experimental instruments: Agilent 1260, EYEL4 rotary evaporator, SB-5200DT ultrasonic cleaner, HWS-26 electric thermostatic water bath, Sartorius BS110S electronic balance;

[0204] Experimental reagents: 95% ethanol, 1,2-dichloroethane, methanol, silica gel, neutral alumina, triptolide;

[0205] Experimental Methods: Pretreatment: Two portions of the extract were transferred to 150 mL round-bottom flasks and concentrated under reduced pressure in a rotary evaporator until completely dry. 50 mL of methanol solution was added, and the mixture was dissolved by sonication. The solution was then transferred to 50 mL centrifuge tubes and centrifuged at 10,000 r / min for 10 min. The supernatant was transferred to an evaporating dish and dried in a water bath at 80 °C to a suitable volume. The triptolide content in the *Tripterygium wilfordii* medicinal material was determined according to the method for determining triptolide content in *Tripterygium wilfordii*.

[0206] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (40:60) was used as the mobile phase; and the detection wavelength was 225 nm. An appropriate amount of triptolide reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 5.3 μg per mL. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed.

[0207] The experimental results are shown in Table 8. Figure 9 and Figure 10 As shown in the experimental results, the extraction rate of triptolide from *Malus kunmingensis* using 50% ethanol alone was 10.0419 μg / g.

[0208] Table 8. Results of Determination of Tripterygium Witchylum Content Extracted Alone from 50% Ethanol in *Malus kunmingensis*

[0209]

[0210] (2) Extraction experiment of 50% ethanol from 4 medicinal materials

[0211] Experimental medicinal materials: Kunming crabapple, epimedium, wolfberry, dodder seed;

[0212] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath, EYEL4 rotary evaporator;

[0213] Experimental Method: Weigh out one prescription amount of medicinal materials: 100g of *Trichosanthes kirilowii*, 100g of *Epimedium brevicornu*, 50g of *Lycium barbarum*, and 50g of *Cuscuta chinensis*. Place the *Trichosanthes kirilowii*, *Epimedium brevicornu*, and *Lycium barbarum* in a 5000mL round-bottom flask, add 3000mL of 50% ethanol, soak for 30 minutes, reflux for 1 hour, and discard the extract. Add 2500mL of 50% ethanol to the residue, reflux for 1 hour, and discard the extract. Add 2500mL of 50% ethanol to the residue again, reflux for 1 hour, and discard the extract. Combine the three extracts and measure their volume and weight. A portion of the extract is used to determine the weight of the dry extract and the content of triptolide, icariin, and betaine. The remaining extract is concentrated under reduced pressure to a paste state for later use. After the dodder seed is pulverized, it is thoroughly mixed with the extract, dried in an oven at 80°C until completely dry, then ground into powder for later use.

[0214] The experimental results are shown in Table 9. After extracting the four herbs of Kunxian with 50% ethanol, the yield of the dry extract was 20.25%.

[0215] Table 9. Results of the 50% ethanol extract of four medicinal herbs from Kunshan.

[0216]

[0217] Tripterygium wilfordii content determination

[0218] Experimental instruments: Agilent 1260, EYEL4 rotary evaporator, SB-5200DT ultrasonic cleaner, HWS-26 electric thermostatic water bath, Sartorius BS110S electronic balance;

[0219] Experimental reagents: 95% ethanol, 1,2-dichloroethane, methanol, silica gel, neutral alumina, triptolide;

[0220] Experimental methods:

[0221] Pretreatment method: Transfer two portions of extract to a 150mL round-bottom flask, concentrate under reduced pressure in a rotary evaporator until completely dry, add 50mL methanol solution, sonicate to dissolve, transfer to a 50mL centrifuge tube, centrifuge at 10000r / min for 10min, transfer the supernatant to an evaporating dish, dry in an 80℃ water bath to an appropriate amount, and determine the triptolide content in the *Tripterygium wilfordii* medicinal material according to the method for determining triptolide content in *Tripterygium wilfordii*.

[0222] Accurately add 50 mL of methanol to a round-bottom flask, weigh it, sonicate (300 W, 40 kHz) for 15 minutes, cool, stopper tightly, and let stand overnight. Accurately pipette 25 mL of the supernatant into an evaporating dish, place it in a 60°C water bath and evaporate to about 3 mL. Add 2.5 g of silica gel (for column chromatography, 200–300 mesh) – neutral alumina (for chromatography, 200–300 mesh) (1:1), mix well, evaporate the methanol in a 60°C water bath while grinding finely, and evaporate the methanol completely. Dry-press the mixture onto a 2 cm inner diameter column containing 2.5 g of a silica gel (for column chromatography, 200–300 mesh) – neutral alumina (for chromatography, 200–300 mesh) (1:1) mixture. Elute with 70 mL of 1,2-dichloroethane, discard the eluent, and then elute with 70 mL of 1,2-dichloroethane solution containing 1.5% ethanol. Collect the eluent, recover the solvent under reduced pressure at 60°C until dry, dissolve in methanol and transfer to a 5 mL volumetric flask, add methanol to the mark, shake well, and filter to obtain the final product.

[0223] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (5% isopropanol) (18:82) was used as the mobile phase; the detection wavelength was 220 nm. An appropriate amount of triptolide reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 7.0 μg per mL. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed.

[0224] The experimental results are shown in Table 10. Figure 11 and Figure 12 As shown, the experimental results indicate that when the four herbs of Kunshan were extracted with 50% ethanol, the extraction rate of triptolide was 10.3008 μg / g.

[0225] Table 10 Results of Determination of Tripterygium Wylene Content in 50% Ethanol Extract from Four Herbs of Kunshan

[0226]

[0227] 1.3 Study on the transfer rate of pure water extraction of Epimedium, Lycium barbarum, and Cuscuta chinensis

[0228] 1.3.1 Study on the transfer rate of Epimedium extract

[0229] (1) Extraction Experiment

[0230] Experimental medicinal material: Epimedium;

[0231] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath;

[0232] Experimental Method: Weigh 200g of Epimedium (for two prescriptions) into a round-bottom flask, add 12 times its volume of drinking water, soak for 30 minutes, reflux for 1 hour, and discard the extract. Add 10 times its volume of drinking water to the residue, reflux for 1 hour, discard the extract, and combine the two extracts. Measure the volume and weight of each extract. A portion of the extract is used for the determination of the weight and content of the dried extract. (The second method is repeated in the original text.)

[0233] The experimental results are shown in Table 11. The results show that the yield of dry extract from Epimedium alone after two extractions was 12.63%, while the yield of dry extract from the residue after a third extraction was 2.09%.

[0234] Table 11 Results of the determination of the dry extract of Epimedium alone

[0235]

[0236] (2) Determination of icariin content

[0237] Experimental instruments: Agilent 1200 high performance liquid chromatograph, SB-5200DT ultrasonic cleaner, Sartorius SQP balance, HG53 moisture analyzer;

[0238] Experimental reagents: Icariin (National Institutes for Food and Drug Control, 110737-201516), 95% ethanol (Guangzhou Chemical Reagent Factory, 202009-1);

[0239] Experimental methods:

[0240] Pretreatment method: Transfer the measured 4 portions of extract to 150mL round-bottom flasks, concentrate under reduced pressure in a rotary evaporator until completely dry, add 25mL of 50% ethanol solution, weigh, sonicate for 1 hour, make up the weight with 50% ethanol, aspirate 15mL of supernatant to a 15mL centrifuge tube, centrifuge at 10000r / min for 10min, filter the supernatant, and inject it into the liquid phase for content determination.

[0241] Chromatographic method: Octadecylsilane-bonded silica gel was used as the stationary phase; acetonitrile-water (30:70) was used as the mobile phase; and the detection wavelength was 270 nm. An appropriate amount of icariin reference standard was accurately weighed and dissolved in methanol to prepare a solution containing 0.1 mg / mL. 10 μL each of the reference solution and the test solution were accurately injected into the liquid chromatograph and analyzed. The theoretical plate number, calculated based on the icariin peak, should not be less than 1500.

[0242] The experimental results are shown in Table 12. Figure 13 and Figure 14 As shown, the icariin extraction rate was 4.0972 mg / g after two extractions of Epimedium alone, and 0.8172 mg / g after a third extraction of the residue.

[0243] Table 12 Results of icariin content determination

[0244]

[0245] 1.3.2 Study on the transfer rate of pure water extraction of wolfberry

[0246] Experimental medicinal material: wolfberry;

[0247] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath;

[0248] Experimental Method: Weigh out the medicinal materials for two prescriptions and extract them separately. Weigh 100g of wolfberry and place it in a round-bottom flask. Add 12 times the volume of drinking water, soak for 30 minutes, and reflux for 1 hour. Pour off the extract. Add 10 times the volume of drinking water to the residue, reflux for 1 hour, and pour off the extract. Combine the two extracts and measure their volume and weight. A portion of the extract is used for the determination of the weight and content of the dried extract. (The second method is repeated in the original text.)

[0249] The experimental results are shown in Table 13. The results show that the yield of dry extract was 64.54% after two water extractions of wolfberry alone, and the yield of dry extract was 1.93% after a third extraction of the residue.

[0250] Table 13 Results of the determination of dry extract from wolfberry alone

[0251]

[0252] 1.3.3 Study on the transfer rate of Cuscuta chinensis extracted with pure water

[0253] Experimental medicinal material: Cuscuta chinensis;

[0254] Experimental instruments: A05-2J-LED-6 balance, TC-15 constant temperature electric heating mantle, HWS-26 electric constant temperature water bath;

[0255] Experimental Method: Weigh 100g of the medicinal materials according to two prescriptions and extract them separately. Place 100g of Cuscuta chinensis in a round-bottom flask, add 12 times its volume of drinking water, soak for 30 minutes, reflux for 1 hour, and pour off the extract. Add 10 times its volume of drinking water to the residue, reflux for 1 hour, and pour off the extract. Combine the two extracts and measure their volume and weight. A portion of the extract is used for the determination of the weight and content of the dried extract. (The second method is repeated in the original text.)

[0256] The experimental results are shown in Table 14. The yield of dry extract from Cuscuta chinensis after two water extractions was 20.83%, while the yield of dry extract from the residue after a third extraction was 3.57%.

[0257] Table 14 Results of the determination of dry extract from Cuscuta chinensis alone

[0258]

[0259] 1.4 Conclusions of the Extraction Process Investigation

[0260] Five experiments were conducted in this small-scale process study: (1) mixed water extraction of four medicinal materials; (2) water extraction of *Tripterygium wilfordii* alone and mixed water extraction of the other three medicinal materials; (3) water extraction of *Epimedium brevicornu*, *Lycium barbarum*, and *Cuscuta chinensis* alone; (4) extraction of *Tripterygium wilfordii* alone with 50% ethanol; and (5) co-extraction of all four medicinal materials with 50% ethanol. The dry extract yield and the extraction rate of triptolide under different extraction processes were mainly investigated. The specific results are as follows:

[0261] (1) Water extraction of four medicinal materials: The yield of dry extract after two extractions was 22.04%, and the extraction rate of triptolide was 5.8110 μg / g. The yield of dry extract after the third extraction was 2.08%, and the extraction rate of triptolide was 1.3578 μg / g. (2) Water extraction of *Trichoderma kunmingense* alone and water extraction of three medicinal materials: The yield of dry extract after two extractions of *Trichoderma kunmingense* alone was 10.42%, and the extraction rate of triptolide was 6.5195 μg / g. The yield of dry extract after the third extraction was 1.08%, and the extraction rate of triptolide was 1.0144 μg / g. The yield of dry extract after two extractions of the remaining three medicinal materials was 26.51%, and the yield of dry extract after the third extraction was 2.85%. (3) Water extraction of Epimedium, Lycium barbarum, and Cuscuta chinensis alone: ​​The yield of dry extract of Epimedium after two extractions was 12.63%, and the extract yield of icariin was 4.0972 mg / g. The yield of dry extract after the third extraction was 2.09%, and the extract yield of icariin was 0.8172 mg / g. The yield of dry extract of Lycium barbarum after two water extractions was 64.54%, and the yield of dry extract after the third extraction was 1.93%. The yield of dry extract of Cuscuta chinensis after two water extractions was 20.83%, and the yield of dry extract after the third extraction was 3.57%. (4) Extraction of Tripterygium wilfordii alone with 50% ethanol: The yield of dry extract of Tripterygium wilfordii with 50% ethanol was 12.68%, and the extract yield of triptolide was 10.0419 μg / g.

[0262] (5) The yield of the dry extract of the four herbs of Kunxian was 20.25% by 50% ethanol extraction, and the extraction rate of triptolide was 10.3008 μg / g.

[0263] The following conclusions can be drawn from the above results: From the perspective of triptolide extraction rate, ethanol extraction is superior to water extraction. However, due to the flammability of ethanol, there are safety hazards during production; therefore, water extraction is preferred in process selection. The results of water extraction show that, compared with single extraction, the extraction rate of triptolide from *Tripterygium wilfordii* was 6.5195 μg / g, which is 12.19% higher than the 5.8110 μg / g of mixed extraction, a significant improvement. Analysis of the extraction and concentration process reveals that *Tripterygium wilfordii* exhibits significant foaming during vacuum concentration, resulting in low efficiency and unsuitability for large-scale production. Therefore, after process investigation and research, it was determined that the original process for *Tripterygium wilfordii*, using water extraction followed by macroporous resin adsorption and elution with 70% ethanol, is mature, stable, and highly efficient.

[0264] The yield of the dry extract from a mixture of Epimedium, Lycium barbarum, and Cuscuta chinensis was 26.51% ± 2.85%. Based on a prescription of 100g Epimedium, 50g Lycium barbarum, and 50g Cuscuta chinensis, the total dry extract yield was 58.72g. If the three herbs were extracted individually by water, the yields of the dry extracts were 12.63% ± 2.09%, 64.54% ± 1.93%, and 20.83% ± 2.85%, respectively. Based on a prescription of 100g Epimedium, 50g Lycium barbarum, and 50g Cuscuta chinensis, the yield was 60.259g, representing a 2.68% increase compared to the mixed extraction. From the extraction process, it can be seen that the extracts of wolfberry and dodder seed alone will produce a lot of foam during the concentration process. Since large-scale production equipment uses vacuum concentration, the individual extraction process is not suitable for large-scale production.

[0265] In summary, through small-scale process research, the original process of preserving *Malus kunmingensis* has been basically determined. Macroporous resin adsorption and elution are used, and a mixed extraction process is used for *Epimedium*, *Lycium barbarum*, and *Cuscuta chinensis*. The next step is to further verify and determine the feasibility of the process through pelleting research.

[0266] 2. Examination of pellet-making process

[0267] 2.1 Small-scale process research

[0268] 2.1.1 Pill dosage form 1

[0269] Experimental medicinal materials: dried powder of Kunming mountain crabapple, epimedium, wolfberry, and dodder seed.

[0270] Experimental Method: Following the original process, *Trichosanthes kirilowii* extract was extracted with pure water, then adsorbed and eluted using macroporous adsorption resin, followed by spray drying to obtain *Trichosanthes kirilowii* dry extract powder. Four portions of 2g *Trichosanthes kirilowii* dry extract powder, 400g *Epimedium brevicornu*, 200g *Lycium barbarum*, and 200g *Cuscuta chinensis* were weighed. The three herbs were placed in a rice cooker, and 9600mL of pure water was added. The mixture was soaked for 30 minutes and then heated for 2 hours for extraction. Pure water was added in two batches (2000mL each) to maintain the liquid level. After extraction, the extract was filtered through a 200-mesh sieve. The residue was then added to 8000mL of pure water and heated for 1 hour for extraction. The extract was then filtered through a 200-mesh sieve. The two filtrates were combined and concentrated using an induction cooker to a fluid extract paste, weighing 0.5829kg.

[0271] The extract was divided into four portions, each weighing approximately 0.1457 kg. Different proportions of starch-microcrystalline cellulose were added to each portion (the excipients were pre-mixed with the dried extract powder of Begonia kunmingensis) and thoroughly mixed to form a soft material.

[0272] ① Prepare 81g of starch-microcrystalline cellulose = 2:1 excipient, premix it with Kunming mountain crabapple dry extract powder, and then mix it thoroughly with extract (145.5g). The soft material is too wet and cannot be shaped. Add starch-microcrystalline cellulose = 2:1 excipient in 3 batches. After mixing, the soft material is too loose, has no toughness and does not stick together, and cannot be made into soft material.

[0273] ② Prepare 80g of starch-microcrystalline cellulose (1:1 ratio) excipient, premix it with Kunming crabapple extract powder, and then thoroughly mix it with the extract (145.4g). The soft material does not take shape. Add the excipient in three batches: 28.1g in the first batch, 8.0g in the second batch, and 7.95g in the third batch. After mixing, the soft material becomes more resilient, but it still cannot be formed into strips.

[0274] ③ Take 80g of starch, premix it with the dried extract powder of Kunming crabapple, and then mix it thoroughly with the extract (145.8g). Since there is no microcrystalline cellulose dispersion, a soft material cannot be made, so this experiment was not carried out.

[0275] ④ Take 137.9g ​​of extract and concentrate it to approximately 89.2g using a water bath. Accurately weigh a portion of the thickened extract for solid content determination, which is 0.4213. Prepare 27.96g of starch-microcrystalline cellulose powder (3:1), premix it with 2g of dried extract powder from *Begonia kunmingensis*, and then mix it with the extract in 5 portions to obtain a suitable soft material suitable for pelleting.

[0276] Experimental results: The final product yielded 67.28g, with 26.50g of remaining soft material, totaling 93.78g. This experiment was equivalent to using 4.345 prescriptions, resulting in a total yield of 21.58g / prescription, with a dosage of 7.19g per dose. Preliminary process analysis determined that the ratio of starch-microcrystalline cellulose powder (3:1), dry extract (calculated based on solid content), and excipients was approximately 1.34:1, which was most suitable for pelleting. Problems: (1) Since this method involves timely mixing of extract, excipients, and dry extract powder to prepare soft material before direct pelleting, the extract cannot be stored for extended periods in large-scale production and must be dried, powdered, and stored as an intermediate product; (2) This pelleting method requires the addition of a large amount of excipients.

[0277] 2.1.2 Pill dosage form 2

[0278] Experimental materials: Kunming crabapple, epimedium, wolfberry, dodder seed;

[0279] Experimental instruments: DT-1002A electronic balance, pilot-scale pulverizer, Sartorius BS110 electronic balance.

[0280] Experimental Method: Weigh 360g of *Malus kunmingensis*, 360g of *Epimedium brevicornu*, and 180g of *Lycium barbarum*. Place them in a rice cooker, add 10800mL of pure water, soak for 30 minutes, heat and extract for 2 hours, filter through a 200-mesh sieve, add 9000mL of pure water, heat and extract for 1 hour, filter through a 200-mesh sieve again, combine the two filtrates, heat and concentrate in an induction cooker to a thick paste, cool, and set aside. Weigh the thick paste to 339.6g. Take two portions of the thick paste and place them in pre-weighed evaporating dishes for determining the solid content ratio. Take 113.40g of the thick paste and mix thoroughly with 66.0g of *Cuscuta chinensis* powder and 33.0g of excipients (starch:microcrystalline cellulose = 3:1). After forming a soft mass, the mass is quite soft, possibly due to the high water content of the thick paste. After slightly cooling, it can be made into pills. Take a portion of the soft mass to make concentrated water pills.

[0281] The remaining thick paste was transferred to an evaporating dish, with a total transfer volume of 188.31g. 38.09g was lost during the transfer, resulting in the transfer of 83.17% of the thick paste. The paste was concentrated in a water bath at 100°C to a volume of 176.11g. 88.05g of this was taken, and 55.44g of dodder seed powder and 27.72g of excipients were added. The mixture was thoroughly mixed to prepare a soft material. A small amount of the thick paste was used for calculating the solid content ratio.

[0282] The experimental results are shown in Table 15. Serial numbers 1 and 2 represent the solid-liquid ratio of the thick paste, and 3 and 4 represent the solid-liquid ratio after the thick paste is concentrated again in a water bath. The total amount of thick paste during the first pill-making was 339.6g. Based on the solid content ratio of 59.62%, the dry paste was 202.46g. If all of it were used for pill-making, 180g of Cuscuta chinensis powder and 90g of excipients would need to be added, resulting in 16.363 prescription amounts, each prescription amount being 28.87g. The dosage for each administration is 9.62g. The dosage of pill form 2 is higher than that of pill form 1.

[0283] Table 15 Results of Solid Content Measurement

[0284]

[0285] 2.1.3 Pill dosage form 3

[0286] Experimental materials: dried powder of *Malus kunmingensis*, epimedium, wolfberry, and dodder seed;

[0287] Experimental instruments: DT-1002A electronic balance, pilot-scale pulverizer, Sartorius BS110 electronic balance.

[0288] Experimental Methods: Weigh 200.0g of Cuscuta chinensis, pulverize it in a pulverizer in the pilot plant, and pass it through a 100-mesh sieve for later use. Weigh 400.0g of Epimedium brevicornu and 200.0g of Lycium barbarum, place them in a rice cooker, add 7200mL of pure water, soak for 30min, heat and extract for 2 hours, filter through a 200-mesh sieve, combine the filtrates, and concentrate them to a thick paste using an induction cooker. The paste weighs 239.68g. A portion of the paste is used for determining the solid content ratio, and approximately 1 / 3 of the paste, 81.2g, is used to prepare a soft material.

[0289] Take 2.67g of dried crabapple extract powder from Kunming Mountain, mix it with 66.66g of dodder seed powder, 33.0g of excipients (starch:microcrystalline cellulose = 3:1), and stir it evenly with 81.2g of thick paste to make a soft material. The soft material has moderate toughness and viscosity and can be used to make pills.

[0290] Experimental results: The solid content of the thick paste was 73.56%, therefore 81.2g of the thick paste contained 59.73g of dry paste. Mixed with 2.67g of dry paste powder, 66.66g of dodder seed, and 33.0g of excipients, a total of 162.06g of solids was obtained. This yielded 6.66 prescriptions, with each prescription containing 26.74g, and a dosage of 8.91g per administration. The advantage of this process is that dodder seed is used as one of the excipients mixed with the thick paste, with a dodder seed to excipient ratio of 2:1, reducing the application of excipients and eliminating the need for binders, thus saving excipient costs. Therefore, this process was selected for further pilot-scale research.

[0291] 2.1.4 Conclusion

[0292] The small-scale experiments above show that dosage form 2 has the highest dosage and is therefore excluded. Dosage form 1 has the lowest dosage, but requires a large amount of excipients, and the mixed extract is not conducive to preservation in large-scale production, which could easily lead to unreasonable production processes. Dosage form 3 has a higher dosage than dosage form 1, but requires less excipients, and the dodder seed is dried and powdered after being mixed with the extract, which is conducive to preservation. The production process is also more reasonable. Therefore, dosage form 3 was ultimately selected as the production process for the pills.

[0293] Example 1

[0294] 1. Pilot production of Kunxian concentrated pills

[0295] Experimental materials: Epimedium (cleaned feed), Lycium barbarum (cleaned feed), Cuscuta chinensis (cleaned feed), dried crabapple extract powder from Kunming Mountain, and refined honey.

[0296] Experimental Method: 4.4 kg and 2.2 kg of cleaned Epimedium and Lycium barbarum were placed in a pilot-scale extraction vessel, and 79.2 L of drinking water was added. The mixture was soaked for 30 minutes, then heated for extraction. The extraction time was started from boiling and continued for 2 hours. The extract was then transferred to a storage tank. The residue was added to 66 L of drinking water and heated for 1 hour. The two extracts were combined and concentrated to a suitable volume in a vacuum concentration vessel. The concentrate was then transferred to a rice cooker and further concentrated to a thick paste, yielding a total of 1970 g of thick paste. One portion of the thick paste was placed in a pre-weighed evaporating dish and dried at 105℃ for 6 hours for determining the solids content.

[0297] Take 2.2 kg of clean dodder seed, crush it and pass it through a 100-mesh sieve for later use. Take 1.1 kg of auxiliary materials (corn starch: microcrystalline cellulose = 1:1) and 52.1 g of dried powder of Kunming crabapple. Mix the above three powders thoroughly in a horizontal mixer, add them to the thick paste, mix thoroughly to form a soft material, and then make it into pellets in a pelleting machine.

[0298] Due to the excessive hardness of the soft material, pelleting failed. The soft material was removed and dried in an oven at 75℃ for approximately 3 hours, then ground into powder, yielding 4.6 kg of powder. This powder was mixed with 2440 mL of pure water and pelleted. The softness of the material was now suitable for pelleting. 140 g of the reserved dodder seed excipient powder was mixed with 28 g of carbon powder, and the mixture was sprinkled into pellets, ultimately yielding 2.09 kg of Kunxian water pellets. An appropriate amount of Kunxian water pellets was taken for pellet weight and moisture content determination.

[0299] Preparation of Kunming Mountain Crabapple Extract Powder: Weigh clean Kunming Mountain Crabapple material and place it in a multi-functional extraction tank. Add drinking water and decoct three times: add 13 times the amount of water for the first decoction, 10 times for the second, and 10 times for the third. Heat to boiling and maintain boiling for 1 hour. Filter the extract through a 100-mesh sieve each time, and store the liquid in a storage tank. Pass the filtrate through a macroporous resin column, and drain the effluent through the drain pipe. After all the extract has been adsorbed by the macroporous resin, wash the resin with drinking water until the effluent is colorless or light-colored, and allow the resin to elute. Add 2.5 times the resin volume of 70% ethanol to the resin column to begin elution. Start collecting the eluent when the color of the effluent becomes significantly darker, and stop collecting the eluent when the ethanol concentration in the effluent is below 30%. Spray dry the eluent using a spray dryer to obtain Kunming Mountain Crabapple Extract Powder.

[0300] Extraction of medicinal materials: Weigh 4.4 kg of cleaned Epimedium and 2.2 kg of cleaned Lycium barbarum, place them in the pilot extraction tank of the water extraction workshop, add 79.2 L of drinking water, soak for 30 minutes, steam heat and reflux extraction for 2 hours, filter, collect the extract into the storage tank, add 66 L of drinking water to the residue, reflux extraction for 1 hour, filter, combine the two extracts, heat and pressurize to concentrate to an appropriate amount, and obtain 2078.4 g of extract. Take about 1 g of extract, divide it into two portions, and use them for the determination of solid content ratio. The experimental results are shown in Table 16.

[0301] Herbal powdering: Weigh 2.2 kg of clean dodder seed material, pulverize it in a pulverizer in the pilot plant, and pass it through a 100-mesh sieve to obtain dodder seed powder;

[0302] Mixing: The dodder seed powder and the extract of medicinal materials are thoroughly mixed and dried in an oven at 80°C for more than 6 hours until the mixture is completely dry. After drying, the mixture is pulverized in a pulverizer in the pilot plant and passed through a 100-mesh sieve to obtain mixed powder. Take 52.1g of dried extract of Kunming crabapple and mix it with the mixture in equal increments to obtain 3802g of mixed powder.

[0303] Pelletizing: Before pelletizing, prepare a honey-water mixture. Take 128.9g of refined honey and mix it with water to make 1032g of honey-water mixture, resulting in a honey content of 12.49%. Weigh out 546g of excipients (corn starch: microcrystalline cellulose = 3:1), and mix them with the powder mixture in equal increments (reserve 300g of the mixture for dusting the pellets). Add the honey-water mixture and mix to form a soft mass. Once the mass is at the appropriate consistency, pelletize it in a pelletizing machine. After pelletizing, use the reserved powder for dusting, and then pelletize the pellets in a pelletizing machine to obtain Kunxian concentrated pellets.

[0304] Drying: The obtained Kunxian concentrated pills were placed in an oven and dried at 60℃ for 5 hours. After drying, a portion of the pills was taken for weight determination, moisture content measurement, and disintegration time determination. The experimental results are shown in Tables 17 and 18. The results show that 3.802 kg of the mixed powder was obtained in this experiment. When mixed with 0.546 kg of excipients and 0.0521 kg of Kunming mountain crabapple extract powder, a total dosage of 4.4001 kg was obtained. This pilot-scale experiment included 200 prescriptions, with a single prescription dosage of 22.0005 g, taken three times daily, with each dose being 7.3335 g, and an average pill weight of 0.1036 g.

[0305] Improvements in this pilot-scale experiment: (1) After the extract is concentrated into a thick paste, it is mixed with dodder seed powder and dried into powder. In actual production, the powder has a longer shelf life than the thick paste. The thick paste is prone to deterioration when left to stand, while the dried powder is more conducive to preservation. (2) The honey content of the honey in this experiment is 12.49%. Since honey has adhesive properties, the amount of excipients can be appropriately reduced. The amount of excipients used in this experiment was reduced to 546g, and the ratio of dodder seed to excipients was 4.02:1, which reduced the use of excipients. (3) After the thick paste is pulverized, it is mixed with dried crabapple extract powder from Kunming Mountain and then directly added to honey for pill making. This reduces the step of heating and drying after mixing with the thick paste, and reduces the loss of the heat-unstable component triptolide. (4) The dosage is further reduced to 7.3335g / time. The difference in pill weight is small, the disintegration time is qualified, and the moisture content is 7.62%. All parameters indicate that this process is the optimal process for Kunming Mountain concentrated pills.

[0306] Table 16 Results of the determination of solid content in the extract

[0307]

[0308] Table 17 Results of Moisture and Disintegration Time Determination of Pilot Products

[0309]

[0310] Table 18 Results of 10 Tests

[0311]

[0312] 2. Determination of the content of key components in Kunxian Concentrated Pills

[0313] (1) Determination of icariin content

[0314] Experimental reagents: 95% ethanol, icariin;

[0315] Experimental materials: pilot-scale product;

[0316] Experimental instruments: DT-1002A electronic balance, Sartorius BS110 electronic balance, KQ-300DE ultrasonic instrument, Waters...

[0317] Arc high-performance liquid chromatograph.

[0318] Experimental procedure: Preparation of standard: Weigh an appropriate amount of icariin accurately to 2.97 mg, and dilute to 25 mL in a volumetric flask with 95% ethanol to prepare a standard solution with a concentration of 118.8 μg / mL. Use after serial dilution with 95% ethanol.

[0319] Sample pretreatment: Take an appropriate amount of the pilot product, grind it, take about 0.5g, make 4 portions, accurately weigh them, add 50mL of 50% ethanol, weigh them, extract by sonication for 1 hour, make up the weight with 50% ethanol, take 15mL of the supernatant into a centrifuge tube, centrifuge at 10000r / min for 10min, take the supernatant, filter it with a 0.22μm microporous membrane, and inject it into the high performance liquid chromatograph for determination.

[0320] Liquid chromatography conditions: Chromatography system: Waters Arc; Column: Agilent XDB C-18, 5μm; Mobile phase: 30% acetonitrile-70% water; Detection wavelength: 270nm; Flow rate: 1mL / min; Column temperature: 30℃; Injection volume: 5μL.

[0321] Experimental results are as follows Figure 15 , Figure 16 As shown in Table 19, the results indicate that the content of icariin in the pilot product was 0.1908 mg / g.

[0322] Table 19 Results of Icariin Determination in Pilot-Scale Products

[0323]

[0324] (2) Determination of Tripterygium wilfordii content

[0325] Experimental reagents: methanol, ethyl acetate, sodium hydroxide, triptolide;

[0326] Experimental materials: pilot-scale product;

[0327] Experimental instruments: DT-1002A electronic balance, Sartorius BS110 electronic balance, DT-5200 ultrasonic instrument, Waters H-Class QDa liquid chromatography-mass spectrometry detector.

[0328] Experimental method: Preparation of standard solution: Tripterygium wilfordii standard prepared by the testing center was serially diluted and set aside.

[0329] Sample pretreatment: Take appropriate amounts of two batches of pilot-scale Kunxian concentrated pills, crush them, and accurately weigh approximately 0.4g each, making four portions. Place each portion in a 50mL Erlenmeyer flask, add 5mL of pure water, heat in a water bath at 80℃ for 5min, accurately add 25mL of ethyl acetate, and ultrasonically extract for 45min. Transfer to a separatory funnel, add 20mL of 3% sodium hydroxide solution, shake well to extract, and allow to stand for layering. Repeat this step twice. Finally, extract once with 20mL of pure water to remove sodium hydroxide. Transfer the entire ethyl acetate layer to a 150mL round-bottom flask, dry under reduced pressure at 60℃, dissolve in methanol solution, and dilute to 5mL in a volumetric flask. Filter using a 0.22μm microporous membrane and inject into a Waters Arc QDa for detection.

[0330] Liquid chromatography conditions: Liquid chromatography system: Waters H-Class UPLC; Column: Waters HSS T3 2.5μm; Detector: Waters QDa; Column temperature: 35℃; Injection volume: 2μL; Mobile phase: Methanol (A) - 0.1% formic acid (B), with gradient elution according to Table 20:

[0331] Table 20 Mobile Phase Proportions

[0332]

[0333] The experimental results are shown in Table 21. Figure 17 and Figure 18 As shown in the results, the triptolide content of the pilot-scale product was 3.204 μg / g. The batch number of the dried extract powder from Kunming Mountain crabapple used was 21092, with a triptolide content of 0.31‰. The amount of dried extract powder used was 52.1g, and the total weight of the pills should be 4.4001kg. Based on the calculation, the triptolide content per gram of pills should be 3.671 μg, while the actual measured content was 3.204 μg / g, resulting in a transfer rate of 87.29%.

[0334] Determination Results of Tripterygium Wilfordii Hook F in Pilot-scale Products in Table 21

[0335]

[0336] Conclusion: As can be seen from Investigation Example 1, a total of 3 kinds of pill-making processes were carried out in this experiment, namely (1) Epimedium, Lycium barbarum, and Cuscuta chinensis were extracted with water and concentrated into a thick paste, which was mixed with the dry paste powder of Tripterygium hypoglaucum Hutch to make Kunxian concentrated pills; (2) Tripterygium hypoglaucum Hutch, Lycium barbarum, and Cuscuta chinensis were extracted with water and concentrated into a thick paste, which was mixed with the powder of Cuscuta chinensis to make Kunxian concentrated pills; (3) Epimedium and Lycium barbarum were extracted with water and concentrated into a thick paste, which was mixed with Cuscuta chinensis, the dry paste powder of Tripterygium hypoglaucum Hutch, and excipients to make Kunxian concentrated pills. If they are all converted into the pilot-scale of 200 prescriptions, the dry paste amount, excipient dosage, and final dosage are as follows in Table 22:

[0337] Table 22 Excipient, Honey Dosage and Dosage of 3 Pill-making Processes

[0338]

[0339] As can be seen from the above table, the excipient dosages of the first 2 processes are relatively large. Only 546 g of excipients are used in Process 3 of the pill dosage form. And from the perspective of the dosage, the dosage of Process 3 of the pill dosage form is lower and the process is more reasonable. Therefore, finally, the improved water pill of Process 3 of the pill dosage form is determined as the pill-making process of the product, and the process flow chart is as Figure 19 shown.

[0340] 3. Pharmacodynamic Research

[0341] 3.1 Animals and Materials

[0342] 3.1.1 Animal Source

[0343] SPF-grade male SD rats, with a body weight of 180.0 g to 220.0 g, were provided by the Guangdong Provincial Center for Medical Laboratory Animals. The license number for the experimental unit to use is: SYXK(Yue)2017-0125. All rats in this experiment were housed in the barrier system of the Experimental Animal Center of Guangdong Pharmaceutical University, with a 12-hour light / 12-hour dark cycle, and provided with pure water and standard food at 24±2°C. All animal experiment procedures have been reviewed and approved by the Ethics Committee of the Animal Experiment Committee of Guangdong Pharmaceutical University, and the ethical approval number is: gdpulacspf2017561.

[0344] 3.1.2 Main Reagents

[0345] Tripterygium wilfordii glycoside tablets (Zhejiang Deende Pharmaceutical Co., Ltd., lot 2111131B); bovine serum albumin (Sigma-Aldrich, USA, lot V900933); lipopolysaccharide (Sigma-Aldrich, USA, lot L2880); castor oil (Tianjin Zhiyuan Chemical Reagent Co., Ltd., lot MB2569); carbon tetrachloride (CCl4, Shanghai Maclean Biochemical Technology Co., Ltd., lot C805332); IL-1β ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., lot MM-0047R1); TNF-α ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., lot MM-0180R2); IL-6 ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., lot MM-0190R1); alanine aminotransferase (ALT / GPT) kit (Nanjing Jiancheng Biotechnology Co., Ltd., lot C009-2-1); aspartate aminotransferase (AST / GOT) kit (Nanjing Jiancheng Biotechnology Co., Ltd., lot... C010-2-1); Protein Quantitative Assay (TP) Kit (Nanjing Jiancheng Biotechnology Co., Ltd., lot A045-2-2); Creatinine (CREA) Kit (Nanjing Jiancheng Biotechnology Co., Ltd., lot C011-2-1); Blood Urea Nitrogen (BUN) Kit (Nanjing Jiancheng Biotechnology Co., Ltd., lot C013-2-1).

[0346] 3.1.3 Main Instruments

[0347] Bio-Rad 680 fully automated microplate reader (BIO-RAD, USA); UV-1780 ultraviolet spectrophotometer (Shimadzu, Japan); 5810 refrigerated centrifuge (Eppendorf, Germany); pathology slide machine (Leica, Germany); optical microscope (Olympus, Japan); Mindray UA-600T urine analyzer (Shenzhen Mindray Bio-Medical Electronics Co., Ltd.).

[0348] 3.2 Experimental Methods

[0349] 3.2.1 Animal Modeling

[0350] After one week of acclimatization, eight SD rats were randomly selected as the control group (BSA) based on body weight, while the remaining rats were used as the model group. Rats in the model group were administered BSA (600 mg / kg) by gavage every two days for 12 weeks, and 0.6 mL of castor oil (containing 0.1 mL of LCl4) was injected subcutaneously once a week for 12 weeks. At weeks 6, 8, and 10, each rat was injected intravenously with 0.05 mg of LPS (lipopolysaccharide). Starting from week 9, rat body weight was measured weekly, and urine was collected in metabolic cages to detect 24-hour proteinuria and observe the model establishment. The control group received an equal volume of physiological saline instead of BSA via gavage, intravenous injection, and subcutaneous injection. At the end of the modeling process (i.e., at the end of the 12th week), 24-hour proteinuria was measured. If the urinary protein in the model group was significantly higher than that in the control group (P<0.05), the model was considered successfully established. After successful modeling, the rats were given BSA (bovine serum albumin, 600 mg / kg) by gavage every 2 days and 0.6 mL castor oil (containing 0.1 mL CCl4) by subcutaneous injection once a week to maintain proteinuria levels.

[0351] 3.2.2 Grouping and Dosing

[0352] Three rats in the modeling group died during the modeling period. Based on the urinary protein content at week 12 of modeling, rats that failed to establish a model were excluded, and the remaining 40 rats were stratified and randomly divided into the IgA model group (Model group), the positive group (Tripterygium wilfordii polyglycoside tablet group (LGT group), the medium and high dose groups of Kunxian capsules, and the low, medium and high dose groups of the pilot product Kunxian concentrated pills, with 8 rats in each group.

[0353] The dosage of Kunxian capsules is 6 capsules / day, or 1.8g / day. Based on an adult weight of 70kg, the dosage of Kunxian capsules is 25.71mg / kg. Using a dose conversion factor of 7 between human and rat body surface area, the dosage for rats is 180.0mg / kg. Therefore, the high-dose group of Kunxian capsules is 180.0mg / kg, and the medium-dose group is 90.0mg / kg. The dosage of the pilot-scale product, Kunxian concentrated pills, is 22.0g / day (divided into 3 doses of 7.3g each). Based on an adult weight of 70kg, the dosage is 314.2mg / kg. Using a dose conversion factor of 7 between human and rat body surface area, the high-dose group is 2199.4mg / kg, the medium-dose group is 1099.7mg / kg, and the low-dose group is 549.8mg / kg. From weeks 13 to 20 of the experiment, each treatment group was administered the corresponding drug once daily by gavage, while the blank control group and IgA model group were given an equal volume of physiological saline (once daily) for 8 weeks. The dosage of Tripterygium wilfordii polyglycoside tablets was 10.0 mg / kg. During the experiment, the 24-hour urinary protein level of each group of rats was continuously monitored. After the administration, relevant organs were weighed and their organ coefficients were calculated; the pathological condition of the kidney tissue of each group of rats was examined; the levels of eleven urinary parameters in each group of rats were detected using an automated urine analyzer; the levels of relevant inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in each group of rats were detected using ELISA; and the levels of relevant biochemical indicators such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Crea) in the liver and kidneys of each group of rats were detected using relevant reagent kits.

[0354] 3.2.3 Observation Items

[0355] (1) Necropsy of laboratory animals

[0356] After the last administration, 24-hour urine samples were collected from each group of animals using metabolic cages for routine urinalysis and 24-hour proteinuria quantification. Animals were weighed, anesthetized, and a midline abdominal incision was made to free the intestines, exposing the abdominal aorta for blood collection. After blood collection, the thoracic and abdominal cavities were exposed, and organs including the thymus, heart, lungs, liver, spleen, kidneys, testes, prostate, and gastrointestinal tract were harvested. These organs were washed with physiological saline, blotted dry with filter paper, and weighed to calculate the organ coefficient (the ratio of organ weight to animal body weight). The kidneys were fixed in 4% paraformaldehyde solution for later use. Histological sections were prepared to observe for pathological changes in the kidneys.

[0357] (2) 24-hour proteinuria and routine urine tests and other biochemical indicators

[0358] Urine was collected from each group of animals in metabolic cages over 24 hours to measure proteinuria levels. After the last administration, urine was collected and routine urinalysis parameters were measured using a urine analyzer: white blood cells (LEU), nitrite (NIT), protein (PRO), pH, occult blood (BLD), specific gravity (SG), ketone bodies (KET), bilirubin (BIL), glucose (GLU), ascorbic acid (VC), and urobilinogen (URO).

[0359] (3) Detection of biochemical indicators such as liver and kidney function

[0360] After the last administration, blood was collected from the abdominal aorta and serum was separated to detect biochemical indicators such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP), creatinine (CREA), and blood urea nitrogen (BUN), which are related to liver and kidney function.

[0361] (4) Serum-related cytokine levels

[0362] TNF-α, IL-1β, and IL-6 levels were measured using an ELISA kit according to the respective instructions.

[0363] (5) Renal pathological tissue pathological examination

[0364] One kidney was harvested, and paraffin sections were stained with hematoxylin and eosin (HE) to observe the pathological changes in renal mesangial cell matrix proliferation.

[0365] 3.2.4 Statistical Processing

[0366] The experimental results were analyzed using SPSS 10.0 (a statistical software package for social sciences). Quantitative data were expressed as mean ± standard deviation (x±s). Pairwise comparisons were performed using t-tests, and ordinal data were analyzed using rank-sum tests. P < 0.05 indicated a statistically significant difference.

[0367] 3.3 Experimental Results

[0368] 3.3.1 General Case

[0369] The control group rats had shiny fur and were in good spirits. The IgA model group rats had dull, sparse fur and reduced activity. Compared with the model group, the rats in different drug administration groups showed some improvement in both fur color and mental state.

[0370] 3.3.2 Effects of the drug administration group on urinary protein in IgA nephropathy rats

[0371] In week 8, the 24-hour proteinuria level in the model group was higher than that in the blank control group, but the difference was not statistically significant (P>0.05). During the modeling period, 24-hour proteinuria did not change significantly in the blank control group, but continued to increase in the model group. In weeks 9, 10, and 11, the 24-hour proteinuria level in the model group was significantly higher than that in the blank control group (P<0.05 or P<0.01). See Table 23.

[0372] At week 12, the 24-hour proteinuria in the model group was significantly higher than that in the blank control group (P<0.01), indicating successful modeling. At week 14, all groups showed significantly lower levels of proteinuria than the model group (P<0.01). From week 16 onwards, the 24-hour proteinuria in all treatment groups was significantly lower than that in the model group (P<0.01). At week 16, the proteinuria in the model group significantly increased, possibly due to kidney injury exceeding the threshold, resulting in a significant increase in proteinuria. Specific results are shown in Table 24. After 8 weeks of administration, i.e., week 20, low, medium, and high doses of Kunxian concentrated pills significantly reduced proteinuria levels in rats. The medium and high doses of Kunxian capsules showed better effects than the Tripterygium wilfordii polyglycoside tablets group. Compared with the same amount of raw Kunxian capsules, the effect of Kunxian concentrated pills was superior to that of the original Kunxian capsule formulation.

[0373] Table 23 Changes in proteinuria in rats of each group 24 hours before drug administration.

[0374]

[0375] Note: Compared with the blank group, * P<0.05, ** P<0.01.

[0376] Table 24 Changes in proteinuria (mg / 24h) in rats of each group after drug administration (24 hours) n=8)

[0377]

[0378] Note: Comparison with model group * P<0.05, ** P<0.01; compared with the same dose of Kunxian capsules group, △△ P<0.01.

[0379] 3.3.3 Effects of the drug administration group on the internal organs of IgA nephropathy rats

[0380] The organ indices of rats in each group were measured, and no statistically significant differences were found in the lung index, heart index, stomach index, testis index, and prostate index (P>0.05).

[0381] The spleen index of the low-dose and medium-dose groups of Kunxian Concentrated Pills was significantly higher than that of the model group (P<0.05 or P<0.01). The thymus index of the low-dose and high-dose groups of Kunxian Concentrated Pills was significantly lower than that of the model group (P<0.05 or P<0.01). The liver index of the medium-dose and high-dose groups was significantly lower than that of the model group (P<0.01). The kidney index of the low-dose, medium-dose, and high-dose groups of Kunxian Concentrated Pills was significantly lower than that of the model group (P<0.01). Regarding the liver and kidney coefficients, the medium-dose Kunxian Concentrated Pills had a smaller effect on the organ coefficients than Kunxian Capsules, indicating that the new dosage form is safer in terms of organ toxicity. See Tables 25 and 26.

[0382] Table 25 Effects of drug administration on organ coefficients in IgA nephropathy rats ( n=8)

[0383]

[0384]

[0385] Note: Comparison with model group * P < 0.05 ** P < 0.01; compared with the same dose of Kunxian capsules, △△ P<0.01.

[0386] Table 26 Effects of drug administration on organ coefficients in IgA nephropathy rats ( n=8

[0387]

[0388] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the same dose KXJ group, △△ P<0.01.

[0389] 3.3.4 Effects of the drug administration group on urinary parameters in IgA nephropathy rats

[0390] There were no statistically significant differences in urinary urobilinogen, ascorbic acid, glucose, occult blood, nitrite, ketones, and specific gravity among the groups (P > 0.05). Urinary urobilinogen, ascorbic acid, and glucose were negative in all groups. The urine pH in the low-dose and medium-dose groups of Kunxian Concentrated Pills was significantly lower than that in the model group (P < 0.05); the urinary white blood cell count in the medium-dose and high-dose groups of Kunxian Concentrated Pills was significantly lower than that in the model group (P < 0.05 or P < 0.01); the bilirubin in the low-dose, medium-dose, and high-dose groups of Kunxian Concentrated Pills was significantly lower than that in the model group (P < 0.01 or P < 0.05); and the urinary protein in the medium-dose and high-dose groups of Kunxian Concentrated Pills was significantly lower than that in the model group (P < 0.01 or P < 0.05). There were no statistically significant differences in any of these indicators between Kunxian Concentrated Pills and Kunxian Capsules at the same dosage (P > 0.05). Detailed results are shown in Table 27.

[0391] Table 27 Effects of drug administration on eleven urinary parameters in IgA nephropathy rats (n=8)

[0392]

[0393]

[0394] 3.3.5 Effects of the drug administration group on liver function in IgA nephropathy rats

[0395] Compared with the model group, the reduction in ALT in the low-dose, medium-dose, and high-dose groups of Kunxian Concentrated Pills was significantly different (P<0.01); the reduction in AST in the low-dose, medium-dose, and high-dose groups of Kunxian Concentrated Pills was also significantly different (P<0.01). At the same dosage, the regulatory effect of Kunxian Concentrated Pills on ALT and AST was significantly better than that of Kunxian Capsules, indicating that the new process has less impact on liver function than the original dosage form and has a better safety profile. Specific results are shown in Table 28.

[0396] Table 28 Effects of drug administration on liver function in IgA nephropathy rats n=8)

[0397]

[0398] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the same dose of Kunxian capsules group, △△ P<0.01.

[0399] 3.3.6 Effects of the drug administration group on renal function in IgA nephropathy rats

[0400] HE staining results are as follows Figure 20As shown, the kidney tissue in the model group exhibited varying degrees of mesangial cell proliferation and increased mesangial matrix, with some showing capillary loop compression, luminal narrowing, and balloon adhesion. The proliferation of glomerular mesangial cells and increased matrix were alleviated in all drug-treated groups.

[0401] Compared with the model group, the BUN (blood urea nitrogen) levels in all treatment groups were significantly reduced (P<0.01). The Cr (creatinine) values ​​in the low-dose, medium-dose, and high-dose groups of Kunxian concentrated pills were all significantly lower than those in the model group (P<0.01). Under the same raw drug dosage, the BUN and CR values ​​in the medium-dose Kunxian concentrated pill group were significantly lower than those in the medium-dose Kunxian capsule group, indicating that the pills were more effective than the original capsule formulation. Specific results are shown in Table 29.

[0402] Table 29 Effects of drug administration on BUN and CR in IgA nephropathy rats ( n=8)

[0403]

[0404] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the same dose of Kunxian capsules group, △△ P<0.01.

[0405] 3.3.7 Effects of the drug administration group on inflammatory markers in IgA nephropathy rats

[0406] Compared with the model group, the low-dose, medium-dose, and high-dose groups of Kunxian concentrated pills all showed a very significant decrease in IL-1β, IL-6, and TNF-α (P<0.01). Under the same raw drug dosage, the high-dose Kunxian concentrated pills had a significantly stronger regulatory effect on IL-1β, IL-6, and TNF-α indicators than Kunxian capsules, while the medium-dose Kunxian concentrated pills were better at regulating IL-1β than Kunxian capsules, indicating that the new pill dosage form is superior to the original dosage form. Specific results are shown in Table 30.

[0407] Table 30 Effects of drug administration on related inflammatory markers in IgA nephropathy rats ( n=8)

[0408]

[0409] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the same dose of Kunxian capsules group, △△ P<0.01.

[0410] 3.4 Conclusion

[0411] IgA nephropathy is the most common type of chronic kidney disease, with abnormal proteinuria as its main clinical manifestation. The animal model used in this study involved oral administration of BSA and CCl4, followed by tail vein injection of LPS. This stimulated the animal's immune response while simultaneously damaging the liver, thus affecting immunoglobulin metabolism and ultimately leading to immunoglobulin accumulation in the kidneys, resulting in kidney damage. This animal model produces significant proteinuria and is the most commonly used rat model for IgA nephropathy in pharmacodynamic studies. Experimental results showed that low, medium, and high doses of the pilot-scale product, Kunxian Concentrated Pills, significantly reduced proteinuria levels in the animals. Simultaneously, the pilot-scale product also significantly reduced blood BUN and Cr levels. These are all commonly used clinical indicators for assessing the progression of IgA nephropathy, indicating that the pilot-scale product has a significant therapeutic effect on IgA nephropathy.

[0412] Histopathological examination revealed varying degrees of mesangial cell proliferation and increased mesangial matrix in the glomeruli of the model group. Some glomeruli showed capillary loop compression, luminal narrowing, and glomerular adhesions, indicating significant kidney damage. After drug administration, these pathological damages were alleviated, further validating the significant efficacy of the drug. The pilot-scale product showed significant effects on liver function indicators AST and ALT, as well as inflammatory markers IL-1β, IL-6, and TNF-α, indicating that the product alleviates kidney damage by reducing the immune system response and relieving inflammation. It also has a certain restorative effect on liver function, mitigating severe liver damage caused by CCl4 and improving the metabolism of immunoglobulins in the body. Furthermore, comparing Kunxian concentrated pills and Kunxian capsules, Kunxian concentrated pills showed stronger regulatory effects on pharmacodynamic indicators, including proteinuria levels, kidney function indicators, and immune factors, compared to Kunxian capsules. However, it had less impact on safety indicators such as organ coefficients and liver function indicators than Kunxian capsules, indicating that the new formulation, Kunxian concentrated pills, indeed achieved reduced toxicity and enhanced efficacy through process improvement. The advantages of the new process are summarized below:

[0413] 1. This process enhances efficacy and reduces toxicity: The new process combines the drugs into a pill form, extracting *Tripterygium wilfordii* separately, and combining the extraction of *Lycium barbarum* and *Epimedium* from the original formula. *Cuscuta chinensis* is now ground into powder for medicinal use. This process retains the water-soluble components of *Epimedium*, *Lycium barbarum*, and *Cuscuta chinensis*, especially *Lycium barbarum* and *Cuscuta chinensis*, which contain a large amount of water-soluble components. These components play a role in regulating immunity and reducing the toxicity of *Tripterygium wilfordii*. Therefore, using this new process to process the drug combination enhances the toxicity reduction and efficacy of the combined medicinal ingredients. In the new process of Kunxian Concentrated Pills, *Epimedium* and *Lycium barbarum* are extracted with pure water, and *Cuscuta chinensis* is ground into powder for medicinal use. Because *Lycium barbarum* and *Cuscuta chinensis* contain a large amount of water-soluble components, they have effects such as regulating immunity, tonifying the liver and kidneys, and reducing toxicity. Compared with the original formulation, retaining these components helps improve efficacy and safety.

[0414] 2. This process offers high production efficiency: The original process involved extracting four medicinal herbs with pure water, separating and purifying the extract using macroporous adsorption resin, collecting the 70% ethanol eluent, concentrating and drying the eluent under reduced pressure to the appropriate amount, and then drying it using a spray dryer to obtain four dry powder extracts, which were then mixed with excipients for granulation. The main limiting factor in production was the use of macroporous resin. Each of the four medicinal herbs required elution time of 13-18 hours, making it the most time-consuming stage in the entire production process. Therefore, the production efficiency of Kunxian capsules was relatively low. Furthermore, resin elution required a large amount of ethanol, necessitating an explosion-proof workshop, resulting in high production costs.

[0415] 3. This process has environmental advantages: The original process for Kunxian Capsules used macroporous resin to purify and enrich the aqueous extracts of wolfberry and dodder seed. This process discharged a large amount of waste liquid containing macromolecular substances, causing the COD of the production wastewater to exceed the standard, reaching 60,000 mg / L, and the wastewater could not be discharged in time. After collection, the wastewater could not be discharged in time due to the high COD, and it needed to be concentrated by membrane separation and collected in multiple batches before it could be discharged. Moreover, the waste liquid was prone to fermentation and deterioration, resulting in poor air quality in the factory. The new process uses pure water to extract epimedium and wolfberry, and the dodder seed is crushed before being used in medicine, retaining the water-soluble components. Wolfberry and dodder seed do not produce wastewater, thus completely eliminating the problem of excessive COD and reducing the sewage discharge pressure of the original process.

[0416] Furthermore, the pharmacological effects of Kunxian Concentrated Pills prepared by this method have been verified: This application uses a combined drug therapy of "bovine serum albumin + carbon tetrachloride + lipopolysaccharide" to establish an IgA nephropathy model. This model closely resembles the symptoms of abnormal proteinuria in clinical IgA nephropathy patients. The pharmacological effects and immune mechanisms were evaluated using proteinuria, creatinine, blood urea nitrogen, cytokines, immunoglobulin levels, and pathological histological examination as indicators. The results showed that Kunxian Concentrated Pills had a significant therapeutic effect on IgA nephropathy rats, reducing proteinuria levels and renal function indicators, alleviating kidney tissue damage, and under the same raw drug dosage, the effect was superior to that of the original Kunxian Capsules.

[0417] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0418] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. A concentrated pill for treating chronic kidney disease, characterized in that, Its preparation materials include the following components: a first extract, a second extract, and pharmaceutically acceptable excipients; The raw materials for preparing the first extract include *Malus kunmingensis*; The raw materials for preparing the second extract include epimedium, wolfberry, and dodder seed; The first extract, the second extract, a mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to form a concentrated drug pill; The second extract includes: dodder seed powder, and an extract prepared by extracting one or more of epimedium, wolfberry and dodder seed individually or in combination.

2. The concentrated pill for treating chronic kidney disease according to claim 1, characterized in that, The active pharmaceutical ingredient includes the following parts by weight: 1-3 parts of *Malus kunmingensis*, 1-3 parts of *Epimedium brevicornu*, 0.5-1.5 parts of *Lycium barbarum*, and 0.5-1.5 parts of *Cuscuta chinensis*.

3. The concentrated pill for treating chronic kidney disease according to claim 1, characterized in that, The active pharmaceutical ingredient includes the following parts by weight: Two portions of *Malus kunmingensis*, two portions of *Epimedium brevicornu*, one portion of *Lycium barbarum*, and one portion of *Cuscuta chinensis*.

4. The concentrated pill for treating chronic kidney disease according to claim 1, characterized in that, The first extract includes dried powder of *Malus kunmingensis*; the second extract includes: an extract prepared by combining the extraction of *Epimedium brevicornu*, *Lycium barbarum* and *Cuscuta chinensis*, or an extract prepared by combining the extraction of *Epimedium brevicornu*, *Lycium barbarum* and *Cuscuta chinensis* separately, or an extract prepared by combining the extraction of *Epimedium brevicornu* and *Lycium barbarum* separately and *Cuscuta chinensis* powder, or an extract prepared by combining the extraction of *Epimedium brevicornu* and *Lycium barbarum* separately and *Cuscuta chinensis* powder.

5. The concentrated pill for treating chronic kidney disease according to claim 4, characterized in that, The second extract comprises: a mixture of powdered dodder seed and extracts of the other two herbs, epimedium and wolfberry, prepared individually or in combination.

6. The concentrated pill for treating chronic kidney disease according to claim 1, characterized in that, Pharmaceutically acceptable excipients include mixtures of starch and microcrystalline cellulose.

7. The concentrated pill for treating chronic kidney disease according to claim 6, characterized in that, In the pharmaceutically acceptable excipients, the mass ratio of starch to microcrystalline cellulose is (1-3):

1.

8. The concentrated pill for treating chronic kidney disease according to claim 1, characterized in that, It is prepared from the effective components in the raw materials mentioned above; Wherein, the Kunming Mountain crabapple is replaced by one or more of the diterpenoids, triterpenoids and alkaloids contained therein; The epimedium is replaced by one or more of the following: epimedium glycoside, epimedium glycoside I, epimedium glycoside II, and epimedium glycoside A contained therein; The dodder seed and wolfberry are replaced by one or more of the flavonoids and polysaccharides contained therein.

9. A method for preparing a concentrated pill for treating chronic kidney disease as described in any one of claims 1-8, characterized in that, Includes the following steps: Provides dried crabapple extract powder from Kunming; The second solvent was used to extract Epimedium and Lycium barbarum separately or in combination, and the resulting extracts were concentrated to prepare an extract. Dodder seeds are crushed to prepare dodder powder; The dodder seed powder, dried crabapple extract powder, extract, mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to form concentrated drug pills; The drug concentrate pill is Kunxian concentrate pill, and the second solvent is water.

10. The method for preparing concentrated pills for treating chronic kidney disease according to claim 9, characterized in that, The combined extraction of Epimedium and Lycium barbarum using a second extraction solvent includes at least one of the following conditions: (1) The extraction method was heating and reflux extraction; (2) The extraction time is 1-3 hours; (3) The volume ratio of the second solvent to the total mass of Epimedium and Lycium barbarum is (10-15) L: 1 kg; (4) The mass ratio of the dodder powder to the pharmaceutically acceptable excipient is (1-3):

1.

11. The method for preparing concentrated pills for treating chronic kidney disease according to claim 4, characterized in that, Includes the following steps: Provides dried crabapple extract powder from Kunming; The second solvent was used to extract Epimedium, Lycium barbarum and Cuscuta chinensis individually or in combination, and the resulting extracts were concentrated to prepare an extract. The Kunming mountain crabapple dried extract powder, extract, mixture of refined honey and water, and pharmaceutically acceptable excipients are mixed to prepare concentrated drug pills; The drug concentrate pill is Kunxian concentrate pill, and the second solvent is water.

12. The concentrated pill for treating chronic kidney disease according to any one of claims 1-8, characterized in that, The preparation method of the dried extract powder of Kunming crabapple includes the following steps: The first solvent was used to extract the *Malus kunmingensis*, and the resulting extract was separated, purified, and dried to prepare *Malus kunmingensis* dry extract powder. The first solvent is water.

13. The method for preparing the concentrated pill for treating chronic kidney disease as described in claim 12, characterized in that, The preparation method of the dried extract powder of Kunming crabapple includes the following steps: Weigh out the crabapple from Kunming Mountain, add water and heat under reflux to extract three times; load the obtained extract into a macroporous resin for adsorption, and after adsorption, elute with 70% ethanol solution; concentrate the obtained eluent under reduced pressure and dry.

14. The use of the concentrated pill of the drug for treating chronic kidney disease according to any one of claims 1-8 in the preparation of a drug for treating chronic kidney disease, rheumatoid arthritis, systemic lupus erythematosus, Crohn's disease, or leprosy reaction.

15. The concentrated pill for treating chronic kidney disease according to any one of claims 1-8, characterized in that, The chronic kidney disease mentioned includes IgA nephropathy.

Images