A composition for delaying progression of chronic kidney disease, and a preparation method and use thereof

Abstract

The application discloses a composition for delaying progression of chronic kidney disease and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine compositions. The composition for delaying progression of chronic kidney disease is obtained by mixing betaine and total flavone extract of pueraria in a mass ratio of 1:0.1-0.2, wherein the content of total flavone in the total flavone extract of pueraria is greater than or equal to 40%. The application realizes the multi-target synergistic effect of betaine and the total flavone extract of pueraria in a specific ratio. The application solves the problems of single drug target and limited curative effect, expands the selection range of treatment of chronic kidney disease, and provides a new drug or health food raw material source with definite curative effect, small adverse reaction and good patient compliance for clinic.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine composition technology, specifically relating to a composition for delaying the progression of chronic kidney disease, its preparation method, and its application. Background Technology

[0002] Chronic kidney disease has become a global public health problem, with end-stage patients requiring dialysis or kidney transplantation to sustain life, imposing a heavy economic burden on patients and society. Renal fibrosis is a common pathological pathway leading to the end-stage of various chronic kidney diseases (CKD), and delaying / reversing renal fibrosis is a core strategy for treating CKD.

[0003] Currently, the main drugs used clinically to slow the progression of CKD include angiotensin-converting enzyme inhibitors / angiotensin II receptor antagonists, sodium-glucose cotransporter 2 inhibitors, and nonsteroidal mineralocorticoid receptor antagonists. While these drugs can reduce proteinuria and slow the decline in renal function to some extent, they still have the following significant limitations in clinical application: Adverse reactions of existing chemotherapy drugs limit their long-term use. For example, ACEI / ARB drugs carry the risk of causing hyperkalemia and transient increases in serum creatinine; some patients even experience acute deterioration of renal function in the early stages of treatment. SGLT2 inhibitors are limited in use in patients with extremely low estimated glomerular filtration rates and carry risks of adverse reactions such as urinary tract infections and ketoacidosis. Furthermore, CKD progression involves the interaction of multiple signaling pathways, including oxidative stress, inflammatory response, apoptosis, and epithelial-mesenchymal transition. Existing chemotherapy drugs mostly target single points, making it difficult to effectively intervene in complex pathological networks and limiting their effectiveness in reversing established renal interstitial fibrosis.

[0004] Therefore, research on the prevention and treatment of CKD using natural products has been increasing in recent years. Extracts of traditional Chinese medicines such as kudzu root, rhubarb, and astragalus have been proven to have renal protective activity. However, existing compositions or compound preparations involving natural products share common problems. Complex composition and unclear material basis: Existing traditional Chinese medicine compound preparations often consist of multiple herbs, resulting in extremely complex chemical compositions and difficulty in identifying the effective component groups. This leads to difficulties in quality control, poor batch-to-batch consistency, and fails to meet the requirements of modern precision medicine and standardized production. Low bioavailability and limited efficacy: Some natural active ingredients (such as flavonoids) have poor water solubility and low oral absorption rates, limiting their full clinical efficacy. Lack of streamlined combinations for multi-target intervention in CKD: While there are existing applications of betaine in liver protection or cardiovascular protection, and the application of kudzu root total flavonoids in improving circulation, there are no reports of combining betaine and kudzu root total flavonoid extracts in a specific ratio specifically for synergistic intervention against the vicious cycle pathway of oxidative stress-inflammation-fibrosis in CKD progression. Summary of the Invention

[0005] The purpose of this section is to outline some aspects of embodiments of the present invention and to briefly describe some preferred embodiments. Simplifications or omissions may be made in this section, as well as in the abstract and title of this application, to avoid obscuring the purpose of these documents; however, such simplifications or omissions should not be construed as limiting the scope of the invention.

[0006] In view of the shortcomings of the prior art, the present invention aims to solve the following specific technical problems: (1) A composition for delaying the progression of CKD with simplified ingredients and controllable quality is provided. It solves the problems of vague ingredients and difficult quality control in existing Chinese medicine compound prescriptions, and only uses two clearly defined ingredients, betaine and kudzu root total flavonoid extract.

[0007] (2) To achieve synergistic effects of two active ingredients at a specific ratio on multiple targets. To solve the problem of single drug targets and limited efficacy.

[0008] (3) Establish a stable preparation method suitable for industrial production. Solve the problems of low extraction rate and low purity of total flavonoids from kudzu root, and provide an extraction and purification process that can effectively enrich the active ingredients of total flavonoids from kudzu root.

[0009] (4) Expand the range of treatment options for chronic kidney disease. Provide a new source of raw materials for drugs or health foods that are effective, have few adverse reactions, and have good patient compliance.

[0010] In extensive screening, the inventors first discovered and focused on the potential synergistic node between betaine as a methyl donor to alleviate endoplasmic reticulum stress and total flavonoids from kudzu root to scavenge oxygen free radicals and inhibit the NF-κB pathway. However, the combination of the two is not a simple functional additive. Through systematic ratio screening studies, it was unexpectedly discovered that only within a very specific mass ratio range (1:0.1–0.2) can the two achieve pharmacodynamic resonance, triggering a powerful anti-inflammatory and antioxidant synergistic signaling network; deviating from this range, not only does the synergistic effect disappear, but antagonism even occurs. This revelation of precise ratios constitutes the core of the inventive work of this invention.

[0011] In view of the problems existing in the above and / or prior art, the present invention is proposed.

[0012] Therefore, the object of the present invention is to overcome the shortcomings of the prior art and provide a composition for delaying the progression of chronic kidney disease. The composition comprises betaine and total flavonoid extract of kudzu root in a mass ratio of 1:0.1 to 0.2.

[0013] As a preferred embodiment of the composition for delaying the progression of chronic kidney disease according to the present invention, wherein the total flavonoid content of the kudzu root total flavonoid extract is ≥40%.

[0014] A preferred embodiment of the method for preparing the composition for delaying the progression of chronic kidney disease according to the present invention includes: extracting dried kudzu root powder and 50%–95% ethanol by heating and reflux 1–3 times, each time for 0.5–2 hours; combining the extracts; and concentrating under reduced pressure to obtain a concentrated solution.

[0015] The concentrated solution was loaded onto a macroporous adsorption resin column. First, water-soluble impurities were removed by elution with 3 to 5 column volumes of deionized water. Then, low-polarity impurities were removed by elution with 2 to 4 column volumes of 10% to 30% ethanol. Finally, desorption elution was performed with 3 to 6 column volumes of 50% to 70% ethanol, and this eluent was collected.

[0016] After recovering the ethanol from the eluent, the extract was concentrated and dried to obtain the total flavonoid extract of kudzu root.

[0017] A mixture of total flavonoid extract from kudzu root and betaine yields a composition for delaying the progression of chronic kidney disease.

[0018] In a preferred embodiment of the method for preparing the composition for delaying the progression of chronic kidney disease according to the present invention, the ethanol used for heating and reflux is 95% ethanol.

[0019] In a preferred embodiment of the method for preparing the composition for delaying the progression of chronic kidney disease according to the present invention, the macroporous adsorption resin column is of type AB-8, D101 or HPD-300.

[0020] As a preferred embodiment of the method for preparing the composition for delaying the progression of chronic kidney disease according to the present invention, the drying includes vacuum drying or spray drying, wherein the temperature of vacuum drying is ≤70°C.

[0021] In a preferred embodiment of the preparation method of the composition for delaying the progression of chronic kidney disease according to the present invention, the relative density of the concentrate measured at 60°C is 1.05 to 1.15.

[0022] The purpose of this invention is to overcome the shortcomings of the prior art and provide an application of a composition in the preparation of a medicament for delaying the progression of chronic kidney disease, wherein the composition for delaying the progression of chronic kidney disease is used as the active ingredient and further comprises pharmaceutically acceptable excipients.

[0023] As a preferred embodiment of the use of the composition described in this invention in the preparation of a medicament for delaying the progression of chronic kidney disease, the excipients include fillers (microcrystalline cellulose, lactose, starch), disintegrants (sodium carboxymethyl starch, crospovidone), binders (povidone K30, hydroxypropyl methylcellulose), and lubricants (magnesium stearate, silicon dioxide).

[0024] As a preferred embodiment of the use of the composition of the present invention in the preparation of a medicament for delaying the progression of chronic kidney disease, wherein the dosage form of the medicament includes tablets, capsules or granules.

[0025] Beneficial effects of this invention: (1) This invention uses the Jin Zhengjun Q-value method to systematically screen the formulation, and for the first time discovered and verified that the synergistic effect of betaine and kudzu root total flavonoid extract has a strict nonlinear characteristic. Only within a narrow ratio window of 1:0.1 to 0.2, the Q value is greater than 1.15, showing a significant synergistic effect. When the ratio deviates from this range, the synergistic effect weakens rapidly and may even turn into antagonism. After locking in the optimal ratio, further pharmacodynamic verification experiments confirmed that the Q value of the combined use of the two on the inhibition of serum creatinine, urea nitrogen, and urinary protein is consistently greater than 1.15, showing a significant synergistic effect. The discovery of this narrow "synergistic window" allows this composition to achieve a multi-dimensional protective effect on renal function far exceeding the simple superposition of single drugs with a simplified composition, achieving better efficacy while reducing the dosage of single drugs and reducing the risk of potential adverse reactions. This nonlinear synergistic phenomenon cannot be predicted by conventional experimental design alone.

[0026] (2) Through systematic pharmacodynamic experiments, this invention comprehensively demonstrated the renal protective effect of the composition from four dimensions: renal function indicators, renal pathology, oxidative stress, and inflammatory factors. Compared with the model control group, each dose group of the composition significantly reduced serum creatinine, blood urea nitrogen, and 24-hour urinary protein excretion in rats with chronic kidney disease (P < 0.05 or P < 0.01), and this effect was dose-dependent. The high-dose group had an effect comparable to or better than enalapril. HE staining of renal tissue showed that the composition group significantly reduced the degree of renal tubular damage, interstitial inflammatory cell infiltration, and fibrosis. At the same time, the composition significantly increased the activity of SOD and GSH-Px in renal tissue (P < 0.05 or P < 0.01) and significantly reduced the serum IL-1β and TNF-α levels (P < 0.05 or P < 0.01). The experimental evidence from the above four dimensions corroborates each other and together constitutes a complete chain of evidence for the renal protective effect of the composition of this invention.

[0027] (3) The composition of this invention is concise, containing only two clearly defined active ingredients, which can be accurately quantified using conventional analytical methods, making it easy to establish and verify quality standards. The raw materials for the preparation of the composition of this invention are widely available, and the preparation process is a classic alcohol extraction-resin purification process, with no risk of high-toxicity reagent residues. Betaine and kudzu root are both natural components with extremely high human tolerance. Animal experiments showed that no obvious signs of liver and kidney toxicity or electrolyte disturbances were observed in the high-dose group, indicating high safety and potential for conversion into food and medicine homology. Attached Figure Description

[0028] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort. Wherein: Figure 1 The serum creatinine levels of rats in different groups of Example 2 and Comparative Examples 1-4 are shown.

[0029] Figure 2 Serum creatinine (A); blood urea nitrogen (B); urinary protein excretion (C); H&E staining of rat kidneys (D) were measured in different groups of rats.

[0030] Figure 3 The superoxide dismutase activity (A) and glutathione peroxidase activity (B) in rat kidney tissues from different groups were compared.

[0031] Figure 4 Serum IL-1β levels (A) and serum TNF-α levels (B) were measured in different groups of rats.

[0032] Figure 5 The values ​​for serum creatinine (A), serum urea nitrogen (B), and urinary protein excretion (C) are for different groups of rats.

[0033] In the above figures, Normal: normal control group; Control: model control group; Betaine: betaine alone group; Pueraria Flavonoids: kudzu root total flavonoid extract alone group; Formulation A~G: formulation groups A~G; Enalapril: enalapril group; low-dose: low-dose group of composition; Moderate-dose: medium-dose group of composition; High-dose: high-dose group of composition; Composition: composition group.

[0034] The data in this invention are expressed as Mean±SEM, compared with the Normal group. # P<0.05, ## P<0.01, ### P<0.001; compared with the Control group, P<0.05, P<0.01, P<0.001. Detailed Implementation

[0035] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0036] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0037] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that is mutually exclusive with other embodiments.

[0038] Unless otherwise specified, all raw materials used in this invention are commercially available.

[0039] Dried rhizome of Pueraria lobata (Willd.) Ohwi, commonly known as wild kudzu (Anhui Bozhou Traditional Chinese Medicine Market); betaine, CAS No.: 107-43-7, purity ≥98% (Chengdu Zhibiao Chemical Biotechnology Co., Ltd.); 0.5% adenine feed (Nantong Trofi Feed Technology Co., Ltd.); enalapril (Yangtze River Pharmaceutical Group Co., Ltd.); microcrystalline cellulose SH-102, sodium carboxymethyl starch, magnesium stearate (Anhui Shanhe Pharmaceutical Excipients Co., Ltd.); 95% ethanol (Sinopharm Chemical Reagent Co., Ltd.); No. 0 capsule shell (Suzhou Capsule Co., Ltd.); lactose Flowlac 100 (Meggle); Microcrystalline cellulose PH102 (Asahi Kasei); Crospovidone XL-10 (Ashland); Povidone K30 (Shinya Pharmaceutical); Sucrose powder (Guangxi Sugar Industry Group); Mannitol (Shandong Tianli Pharmaceutical Co., Ltd.); Citric acid (Shandong Yingxuan Industrial Co., Ltd.); Sweet orange flavor (Aipu Flavor Group Co., Ltd.).

[0040] AB-8 macroporous adsorption resin (Tianjin Bohong Resin Technology Co., Ltd.). Male SD rats, license number: SCXK (Beijing) 2021-0006 (Vital River Laboratory Animal Technology Co., Ltd., Beijing); Detection kits for serum creatinine (Scr), serum urea nitrogen (BUN), urinary microalbumin, superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) (Nanjing Jiancheng Bioengineering Institute); Rat serum interleukin-1β (IL-1β) ELISA detection kit, rat serum tumor necrosis factor-α (TNF-α) ELISA detection kit (Shanghai Jianglai Biotechnology Co., Ltd.).

[0041] Detection method for serum Scr: The sarcosine oxidase method is adopted for detection. After the last administration to the rats, the rats were fasted for 12 hours, blood was collected from the abdominal aorta, and the serum was separated by centrifugation at 3500 r / min for 20 minutes, followed by operations in accordance with the kit instructions.

[0042] Detection method for serum BUN: The urease-glutamate dehydrogenase method is adopted. Take the aforementioned serum, add samples for reaction according to the kit instructions, measure the absorbance at 340 nm with an automatic biochemical analyzer, and calculate the serum urea nitrogen content according to the standard curve.

[0043] Detection method for urinary microalbumin: The immunoturbidimetric method is adopted. A metabolic cage was used to collect 24 h urine of the rats, the urine volume was recorded, the supernatant was taken after centrifugation at 3000 r / min for 10 min, operations were carried out according to the kit instructions, the absorbance was measured at 600 nm with a microplate reader, the urinary microalbumin concentration was calculated according to the standard curve, and the 24 h urinary microalbumin excretion was calculated in combination with the urine volume.

[0044] Renal histopathological examination (HE staining): Rat kidney tissue was fixed in 4% paraformaldehyde for 24 h, subjected to gradient dehydration and paraffin embedding, to prepare 4 μm consecutive sections; the sections were flattened in hot water and dried. After dewaxing, eosin staining was performed. After staining, the sections were dehydrated with absolute ethanol and cleared with xylene, and finally mounted, and the pathological morphology of the kidney tissue was observed under a light microscope. Images were collected and recorded.

[0045] Detection of SOD activity in kidney tissue: The WST-1 method is adopted. Rat kidney tissue was taken to prepare 10% tissue homogenate, centrifuged at 3500 r / min for 10 min, and the supernatant was taken; operations were carried out strictly in accordance with the kit instructions, the absorbance at the corresponding wavelength was measured with a microplate reader, and the SOD activity was calculated.

[0046] Detection of GSH-Px activity in kidney tissue: The colorimetric method is adopted. Take the aforementioned supernatant of kidney tissue homogenate, add samples, incubate, and develop color according to the kit instructions; measure the absorbance with a microplate reader, and calculate the GSH-Px activity according to the standard curve.

[0047] Serum interleukin-1β (IL-1β) content detection: The double antibody sandwich ELISA method was used. Rat serum was taken, and the sample was added, incubated, washed, and colored according to the ELISA kit instructions. The absorbance was measured at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the IL-1β concentration was calculated based on the standard curve.

[0048] Serum tumor necrosis factor-α (TNF-α) content detection: The double antibody sandwich ELISA method was used. Rat serum was collected and the ELISA kit was operated according to the instructions. The absorbance was measured at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the TNF-α concentration was calculated based on the standard curve.

[0049] Example 1: Preparation of total flavonoid extract from kudzu root

[0050] This embodiment provides a method for preparing total flavonoid extract from kudzu root, specifically as follows: 1000g of dried kudzu rhizome was crushed and passed through a 20-mesh sieve. 10L of 95% ethanol was added, and the mixture was extracted twice by reflux, 1.5 hours each time. The filtrates were combined and concentrated under reduced pressure at 60℃ to a relative density of 1.12. The concentrate was passed through a glass chromatography column packed with 2.5kg AB-8 macroporous adsorption resin. Impurities were eluted successively with 10L of deionized water and 8L of 20% ethanol, which were then discarded. Elution was then performed with 15L of 60% ethanol, and the eluent was collected. The eluent was concentrated under reduced pressure at 60℃ to recover the ethanol. The concentrate was then vacuum dried to obtain 32.5g of a yellowish-brown powder. Ultraviolet spectrophotometry (with puerarin as a reference) showed a total flavonoid content of 46.8%, with a yield of 3.25%.

[0051] Example 2

[0052] This embodiment provides a combination for delaying the progression of chronic kidney disease, consisting of betaine and kudzu root total flavonoid extract (prepared in Example 1) in a mass ratio of 1:0.1 to 0.2.

[0053] Comparative Example 1

[0054] The difference between this comparative example and Example 2 is that the mass ratio of betaine and total flavonoid extract of kudzu root was adjusted to 1:0.05, while the other processes were the same as in Example 2, and the composition was prepared.

[0055] Comparative Example 2

[0056] The difference between this comparative example and Example 2 is that the mass ratio of betaine and kudzu root total flavonoid extract was adjusted to 1:0.5, while the other processes were the same as in Example 2, and the composition was prepared.

[0057] Comparative Example 3

[0058] The difference between this comparative example and Example 2 is that the mass ratio of betaine and total flavonoid extract of kudzu root was adjusted to 1:1, while the other processes were the same as in Example 2, and the composition was prepared.

[0059] Comparative Example 4

[0060] The difference between this comparative example and Example 2 is that the mass ratio of betaine and kudzu root total flavonoid extract was adjusted to 20:1, while the other processes were the same as in Example 2, to prepare the composition.

[0061] Ten male Sprague-Dawley SD rats weighing 180–220 g were randomly selected as the normal control group; the remaining rats were fed a diet containing 0.5% adenine to establish a chronic kidney disease (CKD) model. Two weeks after modeling, the model rats were stratified and randomly divided into groups of 10 rats each according to renal function indicators, with one group serving as the model control group without medication. The specific grouping and medication regimens are shown in Table 1 below.

[0062] Table 1 Dosing Regimen

[0063] Except for the normal control group, all animals were continuously given a diet containing 0.5% adenine and administered the corresponding drug by gavage daily for 4 weeks. After the last administration, serum Scr levels were measured in each group of rats, and the interaction type of each administration combination was analyzed using the Jinzhengjun Q-value method. , Among them, E 单药1+单药2 E 单药1 E 单药2 The values ​​represent the effect inhibition rates relative to the model control group in the combination of single drug 1 and single drug 2, the single drug 1 group, and the single drug 2 group, respectively.

[0064] The formula for calculating the inhibition rate E is as follows: E = (mean of model control group - mean of drug treatment group) / mean of model control group × 100%.

[0065] A Q > 1.15 was considered a synergistic effect, a Q between 0.85 and 1.15 was considered an additive effect, and a Q < 0.85 was considered an antagonistic effect. Example 2: Serum Scr levels in rats of Comparative Examples 1-4 were as follows... Figure 1 As shown in Table 2, the experimental results are as follows: Table 2. Effects and synergistic effects of different formulation ratios on serum Scr in CKD rats ( ±s)

[0066] Experiments show that the combined therapeutic effect of betaine and kudzu total flavonoids does not change linearly with dosage. When the ratio is much lower than 1:0.1 or higher than 1:0.2, the synergistic effect disappears rapidly. Even at ratios of 1:0.5 or 1:1, the composition exhibits antagonistic effects, and the therapeutic effect is worse than that of using either alone. This inverted U-shaped dose-response relationship completely negates the common assumption that any combination can enhance efficacy. The composition prepared by this invention using betaine and kudzu total flavonoid extract at a mass ratio of 1:0.1 to 0.2 exponentially surpasses the effect of simple addition. The composition within this range best exerts the multi-target synergistic effect of both and reverses the vicious cycle of oxidative stress-inflammation-fibrosis.

[0067] Example 3

[0068] This embodiment conducted a pharmacodynamic experiment to verify the protective effect of different dosages on a rat model of chronic kidney disease. The mass ratio of betaine to total flavonoid extract of kudzu root was set to 1:0.1, and three dosages (low, medium, and high) were used in the experiment. Specifically:

[0069] Sixty male SD rats, weighing 180–220 g, were randomly divided into six groups of ten rats each: a blank control group, a model control group, a positive control group (enalapril, 10 mg / kg), a low-dose group (betaine 200 mg / kg + kudzu root total flavonoid extract 20 mg / kg), a medium-dose group (betaine 400 mg / kg + kudzu root total flavonoid extract 40 mg / kg), and a high-dose group (betaine 800 mg / kg + kudzu root total flavonoid extract 80 mg / kg). Except for the blank control group, all other groups were fed a diet containing 0.5% adenine to establish a CKD model for six consecutive weeks. From day 1 of modeling, each treatment group was administered the corresponding drug by gavage daily, while the blank control group and the model control group were given an equal volume of distilled water for four consecutive weeks.

[0070] Twenty-four hours after the last administration, 24-hour urine samples were collected from rats to measure urinary protein levels; serum was collected and serum Scr and urinary BUN levels were measured; kidney tissue was taken and hematologic malignancies were observed by HE staining. The experimental results showed that, compared with the model control group, all dose groups of the composition of this invention significantly reduced serum Scr, BUN levels, and urinary protein excretion in rats (P < 0.05 or P < 0.01), and this reduction was dose-dependent. Pathological examination of kidney tissue showed that the composition group rats exhibited significantly reduced renal tubular damage, interstitial inflammatory cell infiltration, and fibrosis. The high-dose group showed effects comparable to or better than the positive control drug enalapril. Figure 2 Experimental results show that the composition of the present invention has a significant protective effect on the kidneys of rats with chronic kidney disease.

[0071] Kidney tissue homogenates were collected from each group of rats, and SOD and GSH-Px activities were measured. The experimental results showed that, compared with the model control group, all dosage groups of the composition of this invention significantly increased SOD and GSH-Px activities in kidney tissue (P < 0.05 or P < 0.01). Figure 3 This indicates that the composition of the present invention has a significant antioxidant effect.

[0072] The serum levels of inflammatory factors IL-1β and TNF-α in rats of each group were detected by ELISA. The results showed that, compared with the model control group, all doses of the composition of this invention significantly reduced serum IL-1β and TNF-α levels (P < 0.05 or P < 0.01). Figure 4 This indicates that the composition of the present invention has significant anti-inflammatory effects.

[0073] Example 4

[0074] This embodiment verifies the synergistic effect. Fifty male SD rats were used to establish a CKD model with a 0.25% adenine diet and randomly divided into 5 groups of 10 rats each: model control group, betaine single treatment group (400 mg / kg), kudzu root total flavonoid extract single treatment group (40 mg / kg), combination group (betaine 400 mg / kg + kudzu root total flavonoid extract 40 mg / kg, i.e., the dose group in Example 3), positive control group (enalapril, 10 mg / kg), and a blank control group of 10 rats. After 4 weeks of administration, serum Scr, BUN, and 24-hour urinary protein excretion were measured in each group of rats. Figure 5 The synergistic effect was evaluated using the Jin Zhengjun Q-value method.

[0075] Table 3 Comparison of serum Scr, BUN and renal function indicators in rats of different groups ( ±s)

[0076] Table 4 Synergistic Effect of Betaine and Total Flavonoids from Pueraria lobata (Jin Zhengjun Q-value Analysis)

[0077] It can be seen that the inhibition rates of betaine alone, kudzu root total flavonoid extract alone, and the combination of the two groups on serum creatinine (Scr) were 28.4%, 23.7%, and 58.6%, respectively, with a calculated Q value of 1.29 (>1.15); the inhibition rates on urinary tract infection (BUN) were 26.3%, 21.9%, and 54.3%, respectively, with a Q value of 1.28 (>1.15); and the inhibition rates on urinary protein were 31.1%, 25.6%, and 61.3%, respectively, with a Q value of 1.26 (>1.15) (Table 4). The results indicate that the combination of betaine and kudzu root total flavonoid extract has a significant synergistic effect.

[0078] Example 5: Preparation of hard capsules (Representative ratio: 1g betaine + 150mg kudzu root total flavonoid extract)

[0079] Prescription: 100g betaine (99% purity), 15g total flavonoid extract of kudzu root obtained in Example 1, 30g microcrystalline cellulose SH-102, 5g sodium carboxymethyl starch, and 1g magnesium stearate.

[0080] Preparation: Add equal amounts of betaine and kudzu root total flavonoid extract sequentially and mix thoroughly. Then add microcrystalline cellulose and sodium carboxymethyl starch and mix for 20 minutes. Prepare a soft mass using 95% ethanol as a wetting agent, granulate through a 20-mesh sieve, dry at 55℃ until moisture content <3%, and granulate again through a 20-mesh sieve. Add magnesium stearate and mix for 5 minutes. Fill into size 0 capsule shells; each capsule has a net weight of approximately 0.4g (containing 100mg betaine and 15mg kudzu root flavonoids).

[0081] Example 6: Preparation of tablets (representative mass ratio of betaine and total flavonoid extract of kudzu root 1:0.2)

[0082] Prescription: 200g betaine, 40g total flavonoid extract of kudzu root, 80g lactose Flowlac 100, 60g microcrystalline cellulose PH102, 12g crospovidone XL-10, 4g magnesium stearate.

[0083] Preparation: Pass the raw materials and excipients through an 80-mesh sieve. Add betaine, kudzu root extract, lactose, microcrystalline cellulose, and crospovidone (8g added) to a wet granulator and mix for 5 minutes. Add an appropriate amount of 5% povidone K30 aqueous solution to prepare a soft mass, granulate through a 24-mesh sieve, and dry in a fluidized bed at 60℃. Sizing the dry granules through a 24-mesh sieve, then add the remaining crospovidone and magnesium stearate and mix. Produce shallow concave tablets, each weighing approximately 0.6g (each tablet contains 200mg betaine and 40mg kudzu root flavonoids).

[0084] Example 7: Preparation of Granules

[0085] Prescription: 100g betaine, 10g total flavonoid extract of kudzu root, 400g sucrose powder, 100g mannitol, 2g citric acid, and appropriate amount of sweet orange flavoring.

[0086] Preparation method: Granulate according to wet granulation process, and package into 5g bags (containing 1g betaine and 100mg kudzu flavonoids).

[0087] In summary, through extensive screening, the inventors discovered and focused for the first time on the potential synergistic node between betaine as a methyl donor to alleviate endoplasmic reticulum stress and total flavonoids from kudzu root to scavenge oxygen free radicals and inhibit the NF-κB pathway. However, the combination of the two is not a simple functional additive. Through systematic ratio screening studies, it was unexpectedly discovered that only within a very specific mass ratio range (1:0.1–0.2) can the two achieve pharmacodynamic resonance, triggering a powerful anti-inflammatory and antioxidant synergistic signaling network; deviating from this range, not only does the synergistic effect disappear, but antagonism even occurs. This revelation of precise ratios constitutes the core of the inventive work of this invention.

[0088] The unexpected synergistic effect of the composition in this invention within a specific, narrow ratio range breaks through conventional linear thinking, achieving a clear synergistic effect and superior efficacy compared to single-drug use. This invention, through systematic ratio screening using the Jin Zhengjun Q-value method, has for the first time discovered and verified a strictly non-linear characteristic in the synergistic effect of betaine and kudzu root total flavonoid extract. Only within a narrow ratio window of 1:0.1–0.2 does the Q-value exceed 1.15, exhibiting a significant synergistic effect; when the ratio deviates from this range, the synergistic effect rapidly weakens, even turning into antagonism. After locking in the optimal ratio, further pharmacodynamic verification experiments confirmed that the Q-values ​​for the inhibitory effects of the two on serum creatinine, blood urea nitrogen, and urinary protein are consistently greater than 1.15, exhibiting a significant synergistic effect. The discovery of this narrow synergistic window allows this composition to achieve a multi-dimensional protective effect on renal function far exceeding the simple superposition of single drugs with a simplified composition, achieving better efficacy while reducing the dosage of single drugs and minimizing potential adverse reaction risks.

[0089] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A composition for delaying the progression of chronic kidney disease, characterized in that: The composition consists of betaine and kudzu root total flavonoid extract in a mass ratio of 1:0.1 to 0.2; The total flavonoid content of the kudzu root total flavonoid extract is ≥40%; The preparation method of the total flavonoid extract from kudzu root includes, Extract kudzu root powder with 50%–95% ethanol by heating and reflux 1–3 times, each time for 0.5–2 hours. Combine the extracts and concentrate under reduced pressure to obtain a concentrated solution. The concentrated solution was loaded onto a macroporous adsorption resin column. First, water-soluble impurities were removed by elution with 3 to 5 column volumes of deionized water. Then, low-polarity impurities were removed by elution with 2 to 4 column volumes of 10% to 30% ethanol. Finally, desorption elution was performed with 3 to 6 column volumes of 50% to 70% ethanol, and this eluent was collected. After recovering the ethanol from the eluent, the extract was concentrated and dried to obtain the total flavonoid extract of kudzu root.

2. The composition for delaying the progression of chronic kidney disease as described in claim 1, characterized in that: The ethanol used for heating and reflux is 95% ethanol.

3. The composition for delaying the progression of chronic kidney disease as described in claim 1, characterized in that: The relative density of the concentrate measured at 60°C was 1.05–1.

15.

4. The composition for delaying the progression of chronic kidney disease as described in claim 1, characterized in that: The macroporous adsorption resin column includes models such as AB-8, D101, or HPD-300.

5. The composition for delaying the progression of chronic kidney disease as described in claim 1, characterized in that: The drying process includes vacuum drying or spray drying, wherein the temperature of vacuum drying is ≤70℃.

6. The use of the composition according to any one of claims 1 to 5 in the preparation of a medicament for delaying the progression of chronic kidney disease, characterized in that: The composition according to any one of claims 1 to 5 is used as the active ingredient, and further includes pharmaceutically acceptable excipients.

7. The use of the composition according to claim 6 in the preparation of a medicament for delaying the progression of chronic kidney disease, characterized in that: The excipients include fillers, disintegrants, binders, and lubricants.

8. The use of the composition according to claim 7 in the preparation of a medicament for delaying the progression of chronic kidney disease, characterized in that: The dosage form of the drug includes tablets, capsules, or granules.

Images