Use of aescin in the preparation of a medicament for the treatment and / or prevention of IgAN

Abstract

The present application relates to the field of medicine, and more particularly to the use of aesculin or verbascoside in the preparation of a medicament for treating and / or preventing IgAN. The present inventors have found that aesculin and / or verbascoside can significantly improve proteinuria caused by IgAN and reduce inflammatory reactions, thereby playing a role in protecting kidney function, and do not have the serious shortcomings of clinically commonly used drugs, indicating that aesculin and / or verbascoside have a strong and low-toxicity effect in treating IgAN. In addition, aesculin and / or verbascoside are a natural compound, low toxicity and easy to be accepted by the human body.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more particularly to the use of lycopene or verbenacin in the preparation of medicaments for the treatment and / or prevention of IgAN. Background Technology

[0002] IgAN (IgA nephropathy) refers to a glomerular disease characterized by the deposition of IgA or predominantly IgA in the glomerular mesangial area. It is currently the most common primary glomerular disease worldwide and a significant cause of end-stage renal disease. The pathogenesis of IgAN is not fully understood, but it is believed to be primarily related to the formation of abnormally glycosylated IgA1 and complement activation. After the production of abnormally glycosylated IgA1, it can bind to abnormal anti-abnormally glycosylated IgA1 antibodies in the body, and simultaneously form immune complexes with the abnormally activated complement system, which deposit in the glomeruli and circulatory system, leading to local inflammation and cell proliferation, ultimately resulting in renal fibrosis. IgAN has an insidious onset, often presenting as asymptomatic hematuria, with or without proteinuria. Some patients present with nephrotic syndrome and varying degrees of renal function impairment. Statistics show that 15%–20% of IgAN patients will develop end-stage renal disease within 10 years, severely impacting their renal function, quality of life, and life expectancy. Currently, treatment methods for IgAN are mainly divided into two types: non-immunosuppressive therapy and immunosuppressive therapy. Commonly used non-immunosuppressive therapies include ACEIs / ARBs, used to control blood pressure and slow the progression of kidney function. Immunosuppressive therapies include glucocorticoids and immunosuppressants (cyclophosphamide, mycophenolate mofetil, azathioprine, and calcineurin inhibitors, etc.). These are all common drugs for treating IgAN, but no single drug can truly and completely control the progression of the disease. For example, ACEIs / ARBs can reduce proteinuria to some extent, but their effect on treating the primary disease is limited; furthermore, some drugs, such as immunosuppressants, have serious side effects—gastrointestinal bleeding, worsening of infections, coagulation disorders, and cardiovascular events. Therefore, there is an urgent need to develop new drugs for treating IgAN. Natural medicines are an important treasure trove in my country. Due to their wide availability, rich components, few adverse reactions, and ease of acceptance by the human body when developed into new drugs, they are attracting increasing attention from researchers both domestically and internationally. Several studies on traditional Chinese medicine formulas for treating IgAN have achieved certain clinical efficacy. For example, modified Shenqi Dihuang Decoction, modified Danggui Buxue Decoction, and Zhuling Decoction combined with Xiaoji Decoction can all reduce proteinuria and improve kidney function. Currently, searching for novel and effective active medicinal substances from natural medicines to treat IgAN has become a research hotspot. Summary of the Invention

[0003] In view of this, the present invention provides the use of verbenacin or arbutin in the preparation of medicaments for the treatment and / or prevention of IgAN. The inventors have discovered that verbenacin and / or arbutin can significantly improve proteinuria caused by IgAN and reduce inflammatory responses, thereby protecting renal function, without the serious drawbacks of commonly used clinical drugs, indicating that verbenacin and / or arbutin have a potent and low-toxicity therapeutic effect on IgAN. Furthermore, verbenacin and / or arbutin, as natural compounds, are low in toxicity and easily tolerated by the human body.

[0004] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0005] This invention provides the use of lycopene or verbenacin in the preparation of medicaments for the treatment and / or prevention of IgAN.

[0006] The present invention also provides compositions for the treatment and / or prevention of IgAN, comprising: honeysuckle glycoside and verbenacin.

[0007] In some embodiments of the present invention, the mass ratio of the honeysuckle glycoside and the verbena glycoside in the above composition is 1:1.

[0008] The present invention also provides the use of the above composition in the preparation of medicaments for the treatment and / or prevention of IgAN.

[0009] In some embodiments of the present invention, in the above applications, the mass ratio of honeysuckle glycoside and verbenacin is 1:1.

[0010] In some embodiments of the present invention, in the above applications, the dosage of the honeysuckle glycoside and the verbena glycoside is 30~90 mg / kg.

[0011] In some embodiments of the present invention, in the above applications, the dosage of the honeysuckle glycoside and the verbena glycoside is 60 mg / kg.

[0012] In some embodiments of the present invention, in the above-described applications, the honeysuckle glycoside and / or the verbena glycoside reduce the number of urinary red blood cells.

[0013] In some embodiments of the present invention, in the above applications, the honeysuckle glycoside and / or the verbena glycoside reduce the content of urinary protein.

[0014] In some embodiments of the present invention, in the above-described applications, the honeysuckle glycoside and / or the verbena glycoside reduce IgA deposition in the glomerular mesangial region.

[0015] In some embodiments of the present invention, the above-described applications of honeysuckle glycoside and / or verbenacin glycoside have any of the following applications;

[0016] (a) Reduces or inhibits the proliferation of mesangial cells in the glomerular mesangial region;

[0017] (b) Reduce or inhibit the deposition of mesangial matrix in the glomerular mesangial area;

[0018] (c) Relieve or reduce pathological damage to kidney tissue.

[0019] The beneficial effects of this invention include:

[0020] (1) This invention provides the use of honeysuckle glycoside and / or verbenacin in the treatment of IgAN: honeysuckle glycoside or verbenacin, when used alone, can significantly reduce urinary hemoglobin and urinary protein, improve renal function (reduce serum creatinine and urea levels), alleviate pathological damage to renal tissue, and reduce IgA deposition in the glomerular mesangial area, thus achieving both symptomatic and root-cause treatment, overcoming the shortcomings of existing technologies such as ACEI / ARB drugs. In particular, when the two are used in combination (mass ratio 1:1), they exhibit a significant synergistic effect, with better efficacy than either drug alone.

[0021] (2) The honeysuckle glycosides and / or verbenacin provided by this invention have the potential to be highly effective in treating IgAN and have significant economic benefits: IgAN is currently the most common glomerular disease worldwide, with the highest incidence in Asian countries. Therefore, the market for IgAN treatment drugs is huge. Monomeric compounds of honeysuckle glycosides or verbenacin, their combinations, or preparations made from pharmaceutically acceptable carriers thereof all have promising applications in treating IgAN. Developed and manufactured in accordance with the national innovative drug approval regulations, they are expected to become a Class 1 highly effective and low-toxicity new drug for treating IgAN with independent intellectual property rights in my country, and their industrialization prospects are clear. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0023] Figure 1 The safety ratings of lycopene, verbenacin, and the combination therapy group are shown.

[0024] Figure 2 Pathological sections of the kidneys of mice in each group are shown (HE staining, 100×).

[0025] Figure 3 Pathological sections of the kidneys of mice in each group (HE staining, 200×) are shown.

[0026] Figure 4 The following are pathological sections of the kidneys of mice in each group (IgA immunofluorescence, 100×).

[0027] Figure 5 The pathological sections of the kidneys of mice in each group are shown (IgA immunofluorescence, 200×). Detailed Implementation

[0028] This invention discloses the use of lycopene or verbenacin in the preparation of medicaments for the treatment and / or prevention of IgAN.

[0029] It should be understood that the expression “one or more of…” individually includes each of the objects described after the expression, as well as various different combinations of two or more of the described objects, unless otherwise understood from the context and usage. The expression “and / or” combined with three or more described objects should be understood to have the same meaning, unless otherwise understood from the context.

[0030] The terms “including,” “having,” or “containing,” including the use of their grammatical synonyms, should generally be understood as open-ended and non-restrictive, for example, not excluding other unstated elements or steps, unless otherwise specifically stated or understood from the context.

[0031] It should be understood that the order of the steps or the order in which certain actions are performed is not important as long as the invention remains operational. Furthermore, two or more steps or actions can be performed simultaneously.

[0032] The use of any and all instances or exemplary language such as “e.g.” or “including” in this document is merely intended to better illustrate the invention and is not intended to limit the scope of the invention unless the claims are made. No language in this specification should be construed as indicating that any unclaimed element is essential to the practice of the invention.

[0033] Furthermore, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Therefore, unless explicitly stated otherwise, it should be understood that all ranges, quantities, values, and percentages used in this disclosure are modified with the word "approximately". Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range.

[0034] The technical solution provided by this invention is:

[0035] 1. Use of verbena or its pharmaceutically acceptable salts in the preparation of medicines for the prevention and / or treatment of IgAN;

[0036] 2. Use of verbenacin and lycopene in combination in the preparation of drugs for the prevention and / or treatment of IgAN;

[0037] Verbenalin, first isolated from plants of the genus *Cornus*, hence its other name, is the first iridoid glycoside discovered in nature. Current research indicates that verbenalin constitutes more than 0.4% of *Verbena officinalis*. It is a white powder at room temperature with a bitter taste. It is almost insoluble in water but soluble in organic solvents such as methanol, ethanol, and DMSO. As a known monomeric compound, verbenalin possesses significant biological activity. It exhibits anti-inflammatory, antioxidant, antitumor, antiviral, antitussive, analgesic, hepatoprotective, neuroprotective, and sleep-promoting effects, and is used clinically to treat edema, dysmenorrhea, amenorrhea, jaundice, and malaria. Pharmacological studies have shown that verbenacin has almost no toxicity to normal epithelial cells, peripheral blood cells, and bone marrow cells. It inhibits inflammatory signaling pathways such as NF-κB, reduces pro-inflammatory factors and chemokines such as TNF-α and IL-6, and simultaneously activates antioxidant pathways such as Nrf2 / ARE, upregulates various endogenous antioxidant enzymes, effectively scavenges reactive oxygen species, and antagonizes oxidative stress damage. It has shown clear protective effects in neurodegenerative diseases, cardiovascular and cerebrovascular diseases, tumors, liver diseases, diabetes, and prostatitis. To date, there are no research reports on the application of verbenacin as a monomeric compound in the preparation of drugs for the treatment of IgAN. The structural formula of verbenacin is as follows:

[0038] .

[0039] Lonicerin is a flavonoid compound extracted from the dried leaves of honeysuckle. It is a pale yellow powder at room temperature. It is almost insoluble in water but soluble in organic solvents such as methanol, ethanol, and DMSO. The structural formula of lonicerin is as follows:

[0040] .

[0041] Lonicera glycosides, as a known monomeric compound, possess significant biological activity. They exhibit anti-inflammatory, anti-arthritis, antioxidant, antifungal, antibacterial, and neuroprotective effects, and have gained importance in clinical applications. Pharmacological studies have shown that lonicera glycosides have almost no effect on normal epithelial cells, peripheral blood cells, and bone marrow cells. They scavenge reactive oxygen species by attenuating NF-κB activity and inhibiting the expression of inflammatory cytokines and chemokines. They play important antioxidant and anti-inflammatory roles in diabetes, cardiovascular disease, inflammatory bowel disease, rheumatoid arthritis and gout, and neurodegenerative diseases. To date, no research reports have been published on the application of lonicera glycosides as a monomeric compound in the preparation of drugs for the treatment of IgAN.

[0042] The present invention also provides the use of pharmaceutical compositions in the preparation of medicaments for the prevention and / or treatment of IgAN, wherein the pharmaceutical composition contains verbenacin, or a geometrical isomer thereof, a pharmaceutically acceptable salt, a solvate or a hydrate.

[0043] In a preferred embodiment of the present invention, one or more pharmaceutically acceptable inert and non-toxic excipients are also included.

[0044] Preferably, the excipient is a carrier, solvent, emulsifier, dispersant, wetting agent, binder, stabilizer, colorant, or fragrance.

[0045] Preferably, the drug is in the form of tablets, capsules, granules, drops, lyophilized material, granules, ointment, or injection.

[0046] Preferably, the verbenacin, or its geometric isomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, is the sole active ingredient.

[0047] In the above technical solution, the IgAN treatment drug composition can be made into tablets, capsules, granules, drops, lyophilized products, ointments or injections by adding common pharmaceutical excipients with honeysuckle glycosides with a purity of 98% or higher.

[0048] The pharmaceutical compositions according to the invention can act systemically and / or locally, and for this purpose can be taken in a suitable manner, such as via oral, parenteral, pulmonary, or nasal routes. The compositions according to the invention can be taken in a form suitable for these routes of administration.

[0049] Suitable for oral administration are administration forms that act and rapidly and / or in an improved manner according to the level of existing technology, and comprise pharmaceutical compositions according to the invention in crystalline and / or amorphous and / or dissolved forms, such as tablets (uncoated or coated tablets, which, for example, have a coating that resists gastric juices or delays dissolution or does not dissolve, upon release of the composition according to the invention), tablets that break rapidly in the mouth, or films, films / lyophilized products, capsules (e.g., hard or soft capsules), sugar-coated tablets, granules, pellets, powders, emulsions, suspensions, aerosols, or solutions.

[0050] Parenteral administration can avoid absorption steps (e.g., intravenous, intra-arterial, intracardiac, intraspinal, intralumbar, or intra-articular) or simultaneously include absorption (e.g., intramuscular, subcutaneous, intradermal, percutaneous, or intraperitoneal). Suitable forms of administration for parenteral administration primarily include formulations for injection and infusion in the form of solutions, suspensions, emulsions, lyophilized products, or sterile powders.

[0051] Another suitable route of administration is, for example, inhaled forms of medication, such as powder inhalers or nebulizers, or nasal forms of medication, such as drops, solutions, or sprays.

[0052] The pharmaceutical composition according to the present invention can be converted into the above-mentioned dosage forms. This can be carried out in a manner known per se by mixing with inert, non-toxic, pharmacologically suitable excipients. These excipients particularly include carriers (such as microcrystalline cellulose, lactose, mannitol, starch), solvents (such as liquid polyethylene glycol), emulsifiers and dispersants or wetting agents (such as sodium dodecyl sulfate, polyoxy sorbitan oleate, propylene glycol), binders (such as polyvinylpyrrolidone), synthetic and natural polymers (such as albumin), stabilizers (such as antioxidants, such as ascorbic acid), colorants (such as inorganic pigments, such as iron oxides) and masking fragrances and odors.

[0053] The effective doses of lonicerin or verbenalin used alone can vary according to the administration route, the age, weight of the patient, the severity of the disease and other relevant factors. When administered orally, the recommended doses of lonicerin or verbenalin are each 40 - 90 mg per dose, 2 - 3 times a day; the recommended doses for parenteral administration are each 10 - 30 mg per dose, once a day. When used in combination, the mass ratio of lonicerin to verbenalin is preferably 1:1, and the total oral dose per time is 80 - 180 mg (including 40 - 90 mg of lonicerin and 40 - 90 mg of verbenalin), 2 - 3 times a day; the total parenteral dose is 20 - 60 mg per dose (including 10 - 30 mg of lonicerin and 10 - 30 mg of verbenalin), once a day.

[0054] In in vivo experiments, an IgAN mouse model was constructed by the method of inducing with xenoproteins of BSA combined with LPS and CCl4 / castor oil. For the first time, a composition of lonicerin and verbenalin was selected as the therapeutic drug. By detecting renal function indexes such as urinary red blood cells, 24-hour urinary protein quantification, serum creatinine (Scr), blood urea nitrogen (BUN), etc., combined with pathological morphological observation of renal tissue HE staining and immunofluorescence detection of the deposition intensity of IgA in the glomerular mesangial area, the preventive and therapeutic effects of lonicerin, verbenalin and their combined use on IgAN were evaluated, and the synergistic q value was calculated by the Jin Zhengjun method, so as to provide an experimental basis for the treatment of IgAN with the composition of lonicerin and verbenalin.

[0055] The materials involved in the present invention:

[0056] 1. Experimental animals

[0057] 50 SPF-grade C57BL / 6JGpt male mice, 6 - 8 weeks old, with a body weight of (25 ± 1.5) g, were provided by Jiangsu Jicui Yakang Biotechnology Co., Ltd. The production license number of experimental animals is (SCXK(Su)2023 - 0009). During the experiment, the temperature of the breeding room was 20 ± 3.2 °C, and the relative humidity was 65 - 75%. They were allowed to eat and drink freely.

[0058] 2. Drugs and reagents:

[0059] 2.1 Medicines

[0060] Verbenin, white powder, content ≥98%, provided by Chengdu Ruifenside Biotechnology Co., Ltd., batch number RDD-M04202511010; Lonicera glycoside, light yellow powder, content ≥98%, provided by Chengdu Ruifenside Biotechnology Co., Ltd., batch number RDD-R06702412017.

[0061] 2.2 Main Reagents

[0062] FITC-labeled rabbit anti-mouse IgA antibody (Sigma-Aldrich, USA), bovine serum albumin (BSA) (Sologne Biotech, Beijing), CCL4 (Aladdin Reagent (Shanghai) Co., Ltd.), castor oil (Aladdin Reagent (Shanghai) Co., Ltd.), DAPI solution (Sologne Biotech, Beijing), 10× EDTA antigen retrieval solution (pH 9.0) (Hubei Bios Biotech Co., Ltd.), fluorescent mounting medium (Southern Biotech), urea nitrogen assay kit, creatinine assay kit, and urine protein quantification test kit were all purchased from Nanjing Jiancheng Bioengineering Institute, and PBS buffer (Beyotime Biotech, Shanghai).

[0063] 2.3 Main Instruments

[0064] Dehydrator (Changzhou Zhongwei Electronic Instruments Co., Ltd.), embedding machine (Changzhou Zhongwei Electronic Instruments Co., Ltd.), pathology slide machine (Thermo Fisher Scientific), fluorescence microscope (Olympus Ltd.), anti-detachment slide (Hubei Bios Biotechnology Co., Ltd.), imaging system (3D HISTECH).

[0065] The preparation examples provided in this invention are merely illustrative; other dosage forms are also included in the technical solutions of this invention.

[0066] In the preparation examples, embodiments, and effect examples of this invention, all raw materials and reagents used are commercially available.

[0067] The present invention will be further illustrated below with reference to the embodiments:

[0068] Preparation Example 1 (Compound Capsule)

[0069] Take 450 g of honeysuckle glycoside monomer compound and 450 g of verbenacin monomer compound, add 300 g of medicinal starch, mix thoroughly, and make 1000 capsules, each weighing 1.2 g and containing 450 mg of honeysuckle glycoside and 450 mg of verbenacin. A 60 kg adult should take 8 capsules at a time.

[0070] Preparation Example 2 (Compound Tablets)

[0071] Take 450 g of honeysuckle glycoside monomer compound and 450 g of verbenatin monomer compound, add 300 g of medicinal starch, mix thoroughly, and compress into 1000 tablets, each weighing 1.2 g, containing 450 mg of honeysuckle glycoside and 450 mg of verbenatin. A 60 kg adult should take 8 tablets at a time.

[0072] Example 1: Modeling and Drug Administration

[0073] Fifty SPF-grade male C57BL / 6JGpt mice were weighed, numbered, and randomly divided into five groups: a normal control group, an IgAN model group, a verbenacin dosage group (60 mg / kg), a lycopene dosage group (60 mg / kg), and a combined treatment group (verbenin 60 mg / kg + lycopene 60 mg / kg), with 10 mice in each group. Except for the normal control group, all mice underwent IgAN induction using a heterologous protein induction method with BSA + LPS + CCl4 + castor oil. The normal control group mice were given the same volume of physiological saline as the model group and the treatment groups via gavage or injection, but were not treated with bovine serum albumin (BSA), lipopolysaccharide (LPS), carbon tetrachloride (CCl4), or other modeling drugs, and maintained normal diet and feeding conditions. The specific procedures are as follows: BSA 400mg / kg was administered by gavage every other day for 8 weeks, and CCl4 0.1mL + castor oil 0.4mL was injected subcutaneously once a week for a total of 9 weeks. In the 6th week, 0.05mg LPS was injected via the tail vein. Observation continued until the end of the 12th week.

[0074] Administration method: Drug administration began after model establishment. Verbenin and honeysuckle were suspended in sodium carboxymethyl cellulose (CMC) and administered by gavage. Mice in the normal control and model groups were administered physiological saline by gavage (10 mL / kg) daily; the verbenin group was administered verbenin suspension by gavage (60 mg / kg); the honeysuckle group was administered honeysuckle suspension by gavage (60 mg / kg); and the combined treatment group was administered verbenin + honeysuckle suspension by gavage (60 mg / kg each). The administration volume was 10 mL / kg, once daily for two consecutive weeks.

[0075] 2. Specimen collection and detection of blood and urine biochemical indicators

[0076] Urine collection: One day before the end of the experiment, morning urine and 24-hour urine were collected using a metabolic cage, and the 24-hour urine volume was recorded. Urine sediment was examined under a microscope to determine the urine red blood cell count; urine protein concentration was determined using a urine protein detection kit (CCB method), and the 24-hour urine protein quantification was calculated.

[0077] Blood collection and serum preparation: After the drug intervention, the patient was anesthetized by intraperitoneal injection of 2% sodium pentobarbital, blood was collected from the eyeball, and serum was separated (3000 r / min, 15 min). Serum creatinine and blood urea nitrogen levels were then measured.

[0078] Kidney tissue collection: After drug intervention, the patient was anesthetized by intraperitoneal injection of 2% sodium pentobarbital, and the left kidney tissue was collected. The tissue was fixed with 4% paraformaldehyde to form a paraffin block and then frozen at -80°C for later use.

[0079] 3. Histopathological observation

[0080] Kidney tissue was dehydrated, dewaxed, embedded, and stained with hematoxylin and eosin (HE). Pathological changes in mouse kidneys were observed under a microscope.

[0081] 4. Immunofluorescence observation of kidney tissue

[0082] After dewaxing, hydration, and EDTA (pH 9.0) autoclaving for antigen retrieval, sections were blocked with 10% normal donkey serum at room temperature for 30 minutes. Primary antibody (IgA-FITC, 1:100) was then added and incubated overnight at 4°C. The next day, sections were washed with TBST, incubated with the corresponding secondary antibody (Alexa Fluor® 594 donkey anti-rabbit, 1:600, 37°C for 45 minutes), counterstained with DAPI nuclei, and finally mounted with anti-fluorescence quenching mounting medium. All sections were observed and images were acquired under a fluorescence microscope (Olympus BX53).

[0083] 5. Statistical methods

[0084] All experimental data were obtained based on three independent replicate experiments. Statistical analysis and graphing were performed using GraphPad Prism 9.0 and SPSS software. One-way ANOVA was used for comparisons among multiple groups. A p-value < 0.05 was considered statistically significant.

[0085] Example of effect

[0086] 1. Effects of verbenacin, lycopene, and their combined use on urinary erythrocytes and urinary protein in mice.

[0087] As shown in Table 1, experimental mice were treated by gavage with lycopene, verbenacin, or a combination of both, and urine was collected for urinary red blood cell count and 24-hour urinary protein quantification. The results showed that the urinary red blood cell count and 24-hour urinary protein quantification in the model group were significantly higher than those in the normal control group (P<0.05); the above indicators in each treatment group were significantly lower than those in the model group (P<0.05), and the combined drug group was significantly more effective than any single drug group (P<0.05).

[0088] Table 1 Comparison of urinary erythrocyte and urinary protein levels in mice of different groups

[0089]

[0090] Note:* Compared with the control group, P<0.05; # Compared with the model group, P < 0.05;

[0091] 2. Effects of verbenacin, lycopene, and their combination on renal function indicators Scr and BUN in mice.

[0092] As shown in Table 2, the serum Scr and BUN levels in the model group were significantly higher than those in the normal control group (P<0.05). The serum Scr and BUN levels in each treatment group were significantly lower than those in the model group (P<0.05). Among them, the combined drug group showed the best effect on improving renal function, which was significantly better than any single drug group (P<0.05), and its BUN and Scr levels were no longer statistically different from those in the normal control group.

[0093] Table 2 Comparison of renal function-related indicators among different groups of mice

[0094]

[0095] Note: * Compared with the control group, P<0.05; # Compared with the model group, P < 0.05;

[0096] 3. Effects of verbenacin and lycopene on pathological changes in mouse kidney tissue

[0097] like Figure 2 and Figure 3 As shown, under light microscopy, most glomerular mesangial cells in the model group exhibited severe proliferation, increased matrix, focal segmental sclerosis, and multifocal tubular atrophy. In both verbenacin and honeysuckle-based treatment groups, mesangial cell and matrix proliferation were reduced, with occasional segmental sclerosis and glomerular adhesions. Pathological changes indicated that honeysuckle-based treatment was less effective than verbenacin in treating IgA nephropathy. The combined treatment of both showed better results.

[0098] 4. Effects of verbenacin and lycopene on IgA immunofluorescence in renal tissue

[0099] like Figure 4 and Figure 5 As shown, no IgA deposition was observed in the glomerular mesangial area of ​​the normal control group, while strong, clumpy, green, intense IgA fluorescence or granular, moderate-intensity IgA fluorescence was observed in the glomerular mesangial area of ​​the model group. The IgA fluorescence intensity in the glomerular mesangial area of ​​mice in each drug intervention group was significantly weaker than that in the model group (P<0.05). Lonicerin was less effective than verbenatin in improving IgA deposition in IgA nephropathy. The synergistic effect of both treatments was better.

[0100] Table 3

[0101]

[0102] The synergistic effect q value of the combined inhibition rate of IgA by honeysuckle glycoside and verbenatin was calculated by the Jin Zhengjun method. The q value was 1.23, indicating that the combined drug treatment has a synergistic effect.

[0103] In summary, continuous gavage administration of verbenacin to IgAN mice for two weeks significantly reduced urinary hemoglobin and 24-hour urinary protein levels. Verbenin significantly reduced serum Scr and BUN levels in IgAN mice and improved renal function. Renal histopathology showed that both verbenacin and honeysuckle glycoside treatment reduced mesangial cell proliferation and mesangial matrix proliferation in the glomerular mesangial area of ​​IgAN mice. Immunofluorescence observation of renal tissue indicated a significant decrease in IgA fluorescence intensity in the glomerular mesangial area after treatment with both verbenacin and honeysuckle glycoside. Both verbenacin and honeysuckle glycoside improved renal function in IgAN mice, reduced urinary hemoglobin and protein, and alleviated renal histopathological damage. From a therapeutic perspective, verbenacin is more effective than honeysuckle glycoside in treating IgA nephropathy, and the two may have a synergistic effect.

[0104] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of lycopene or verbenacin in the preparation of drugs for the treatment and / or prevention of IgAN.

2. A composition for the treatment and / or prevention of IgAN, characterized in that, include: Honeysuckle glycoside and verbenacin.

3. The composition according to claim 2, characterized in that, The mass ratio of the honeysuckle glycoside and the verbena glycoside is 1:

1.

4. Use of the composition according to claim 2 or 3 in the preparation of a medicament for treating and / or preventing IgAN.

5. The application as described in claim 4, characterized in that, The application rate of the honeysuckle glycoside and the verbenacin glycoside is 30~90 mg / kg.

6. The application as described in claim 5, characterized in that, The application rate of the honeysuckle glycoside and the verbenacin glycoside is 60 mg / kg.

7. The application as described in any one of claims 4 to 6, characterized in that, The honeysuckle glycoside and / or the verbena glycoside reduce the number of red blood cells in the urine.

8. The application as described in any one of claims 4 to 7, characterized in that, The honeysuckle glycoside and / or verbenacin reduce the content of urinary protein.

9. The application as described in any one of claims 4 to 8, characterized in that, The honeysuckle glycoside and / or verbenacin reduce IgA deposition in the glomerular mesangial region.

10. The application as described in any one of claims 4 to 9, characterized in that, The honeysuckle glycoside and / or the verbena glycoside have any of the following applications; (a) Reduces or inhibits the proliferation of mesangial cells in the glomerular mesangial region; (b) Reduce or inhibit the deposition of mesangial matrix in the glomerular mesangial area; (c) Relieve or reduce pathological damage to kidney tissue.

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