Application of hispidulin in preparation of medicine for treating lupus nephritis

Abstract

The invention relates to application of hispidulin or pharmaceutically acceptable salts, esters and solvates thereof in preparation of medicines for treating lupus nephritis. The invention develops a new application of the hispidulin, and proves the treatment effect of the hispidulin on lupus nephritis. Meanwhile, the invention verifies that the hispidulin can obviously improve the renal function indexes (including reducing urine protein, blood urea nitrogen and serum creatinine, increasing serum albumin and improving lipid metabolism) of lupus nephritis, can relieve renal pathological injuries (including reducing glomerular mesangial hyperplasia, basilar membrane thickening, renal tubule dilatation and interstitial inflammation infiltration), and can improve the renal function indexes of lupus nephritis. Kidney immune complex deposition can be reduced (including reduction of deposition of kidney IgG and complement C3), meanwhile, it is found for the first time that angiogenin-like protein 4 (Angptl4) is a key action target for treating lupus nephritis by means of the homospidulin, and the homospidulin plays a kidney protection role by down-regulating Angptl4 expression.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to application of high plantain (Hispidulin) or a pharmaceutically acceptable salt, ester or solvate thereof in preparation of a drug for treating lupus nephritis. BACKGROUND

[0002] Systemic Lupus Erythematosus (SLE) is an autoimmune disease that involves multiple organs of the whole body, and the disease condition alternates between repeated attacks and remission. Among them, the kidney is the most common organ involved in the pathogenesis of SLE. Lupus nephritis (LN) is an immune complex nephritis caused by systemic lupus erythematosus involving the kidney, and the pathological changes mainly include mesangial cell proliferation, increased mesangial matrix, and obvious inflammatory cell infiltration in the glomerulus and renal interstitium; there are also crescents, glomerular capillary hyperplasia, renal interstitial sclerosis, and renal tubular atrophy, etc. LN is the main cause of death in SLE patients.

[0003] As one of the most serious complications of SLE, 10%-15% of LN patients will eventually develop end-stage renal disease.

[0004] The existing treatment regimen for lupus nephritis has the following shortcomings:

[0005] (1) Many adverse reactions: long-term use of glucocorticoids can increase the risk of infection, osteoporosis, glucose metabolism disorder, and serious side effects such as gastrointestinal ulcers; immunosuppressive agents can cause bone marrow suppression, liver and kidney function damage, infection and other complications.

[0006] (2) Uncertain efficacy: some patients do not respond well to existing treatment regimens, and there are refractory or recurrent lupus nephritis.

[0007] (3) Expensive: the high price of biological agents limits their widespread use in clinical practice.

[0008] Therefore, there is an urgent need to develop new, safe and effective, and kidney-protective drugs for the treatment of lupus nephritis. Natural flavonoids provide a new research direction for the treatment of lupus nephritis due to their good safety, multi-target effect and fewer adverse reactions.

[0009] Hispidulin is a natural flavonoid compound with a molecular formula of C 16 H 12 O6 and a molecular weight of 300.26. Hispidulin mainly exists in the leaves, seeds and whole grass of the plantain family such as plantain. Previous studies suggest that it has significant anti-inflammatory, antioxidant and anti-tumor pharmacological effects.

[0010] In the cardiovascular field, studies have confirmed that psyllium has therapeutic effects on myocardial hypertrophy and heart failure. CN112156091A discloses the application of psyllium in the preparation of drugs for the treatment and / or prevention of cardiovascular diseases, confirming that psyllium can alleviate myocardial hypertrophy and heart failure by improving mitochondrial function and inhibiting oxidative stress.

[0011] In the field of nephrology, research has found that psyllium husk can improve acute kidney injury in mice through inflammation, oxidative stress, and apoptosis.

[0012] However, to date, there have been no reports of the use of psyllium husk in the preparation of drugs for the treatment of lupus nephritis. Summary of the Invention

[0013] In view of the shortcomings of the prior art, the purpose of this invention is to provide the use of psyllium oleoresin or its pharmaceutically acceptable salts, esters, and solvates in the preparation of medicaments for treating lupus nephritis.

[0014] To achieve this objective, the present invention adopts the following technical solution:

[0015] In a first aspect, the present invention provides the use of psyllium oleraceum or its pharmaceutically acceptable salts, esters, or solvates in the preparation of medicaments for treating lupus nephritis.

[0016] This invention develops a new use for psyllium glycosides, demonstrating its therapeutic effect on lupus nephritis and filling a gap in the application of psyllium glycosides in the field of autoimmune kidney diseases. Simultaneously, this invention verifies that psyllium glycosides can significantly improve renal function indicators in lupus nephritis (including reducing proteinuria, blood urea nitrogen, and serum creatinine, increasing serum albumin, and improving lipid metabolism), alleviate renal pathological damage (including reducing glomerular mesangial proliferation, basement membrane thickening, renal tubular dilation, and interstitial inflammatory infiltration), and reduce renal immune complex deposition (including reducing renal IgG and complement C3 deposition). Furthermore, it is the first time that Angptl4 has been discovered as a key target for psyllium glycosides in the treatment of lupus nephritis, and psyllium glycosides exert its renal protective effect by downregulating Angptl4 expression.

[0017] In this invention, purpuricin can be used as a single active ingredient with a well-defined structure to treat lupus nephritis. This is fundamentally different from a mixture of multiple components such as plantain and plantain seeds, as well as purpuricin, which is a prodrug-like substance that needs to be hydrolyzed in vivo to be converted into purpuricin before it can exert its effects.

[0018] Preferably, the drug further contains pharmaceutically acceptable excipients.

[0019] Preferably, the pharmaceutically acceptable excipients include any one or a combination of at least two of the following: fillers, binders, wetting agents, disintegrants, solubilizers, osmotic pressure regulators, coating materials, colorants, pH adjusters, antioxidants, or antibacterial agents.

[0020] Preferably, the dosage form of the drug includes any one of dispersible tablets, sustained-release tablets, effervescent tablets, enteric-coated tablets, sustained-release capsules, enteric-coated capsules, granules, injections, pellets, or solutions.

[0021] Secondly, the present invention provides the use of psyllium argentide or its pharmaceutically acceptable salts, esters, and solvates in the preparation of regulators of renal function-related serum biochemical indicators.

[0022] Preferably, the renal function-related serum biochemical index regulator includes any one or a combination of at least two of the following: regulators that lower blood urea nitrogen levels, regulators that lower creatinine levels, regulators that lower cholesterol levels, or regulators that increase albumin levels.

[0023] Thirdly, the present invention provides the use of psyllium argentide or its pharmaceutically acceptable salts, esters, and solvates in the preparation of medicaments for improving renal histopathological damage.

[0024] Preferably, the renal histopathological damage includes any one or a combination of at least two of the following: glomerular lesions, tubulointerstitial damage, basement membrane thickening, or mesangial matrix deposition.

[0025] Preferably, the drug further contains pharmaceutically acceptable excipients.

[0026] Preferably, the pharmaceutically acceptable excipients include any one or a combination of at least two of the following: fillers, binders, wetting agents, disintegrants, solubilizers, osmotic pressure regulators, coating materials, colorants, pH adjusters, antioxidants, or antibacterial agents.

[0027] Preferably, the dosage form of the drug includes any one of dispersible tablets, sustained-release tablets, effervescent tablets, enteric-coated tablets, sustained-release capsules, enteric-coated capsules, granules, injections, pellets, or solutions.

[0028] Fourthly, the present invention provides the use of psyllium argentide or its pharmaceutically acceptable salts, esters, or solvates in the preparation of immune complex deposition inhibitors.

[0029] Preferably, the immune complex deposition inhibitor includes an IgG deposition inhibitor and / or a C3 deposition inhibitor.

[0030] Fifthly, the present invention provides the use of psyllium argentide or its pharmaceutically acceptable salts, esters, or solvates in the preparation of angiopoietin-like protein expression inhibitors.

[0031] Preferably, the angiopoietin-like protein expression inhibitor includes an angiopoietin-like protein 4 expression inhibitor.

[0032] This invention demonstrates, using a mouse model of nephrotoxic serum nephritis, that psyllium tumefaciens has an inhibitory effect on the occurrence / development of lupus nephritis. That is, psyllium tumefaciens can be formulated into a drug for the treatment of lupus nephritis, or into a simple experimental preparation for the purpose of exploring serum biochemical indicators, renal tissue pathological damage, and other related fields of lupus nephritis.

[0033] The above-mentioned applications protected by this invention are not intended to eliminate the cause or lesion, that is, for the purpose of non-disease treatment.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] This invention develops a new use for psyllium glycosides, demonstrating its therapeutic effect on lupus nephritis and filling a gap in the application of psyllium glycosides in the field of autoimmune kidney diseases. Simultaneously, this invention verifies that psyllium glycosides can significantly improve renal function indicators in lupus nephritis (including reducing proteinuria, blood urea nitrogen, and serum creatinine, increasing serum albumin, and improving lipid metabolism), alleviate renal pathological damage (including reducing glomerular mesangial proliferation, basement membrane thickening, renal tubular dilation, and interstitial inflammatory infiltration), and reduce renal immune complex deposition (including reducing renal IgG and complement C3 deposition). Furthermore, it is the first time that Angptl4 has been discovered as a key target for psyllium glycosides in the treatment of lupus nephritis, and psyllium glycosides exert its renal protective effect by downregulating Angptl4 expression. Attached Figure Description

[0036] Figure 1 This is the experimental design flowchart for Example 1.

[0037] Figure 2 This is a comparison chart of kidney function indicators in mice from different experimental groups.

[0038] Figure 3 This is a comparison chart of liver function indicators in mice from different experimental groups.

[0039] Figure 4 These are comparison images of PAS staining in mice from different experimental groups.

[0040] Figure 5 These are comparison images of immunofluorescence staining of mice in different experimental groups.

[0041] Figure 6 This is a comparison of the relative mRNA expression levels of Angptl4 in the kidney tissues of mice in each experimental group. Detailed Implementation

[0042] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0043] Example 1

[0044] This embodiment verifies the therapeutic effect of psyllium husk on a mouse model of lupus nephritis.

[0045] (1) Experimental materials:

[0046] (1.1) Experimental animals: SPF-grade male C57BL / 6 mice, 6-8 weeks old, weighing 18-20 g, were purchased from Guangdong Yaokang Biotechnology Co., Ltd. and housed in an SPF-grade animal room with free access to water and food. The housing environment was: room temperature 22±2℃, relative humidity 50-60%, and 12-hour light-dark alternation.

[0047] (1.2) High-quality carp.

[0048] (2) Experimental method:

[0049] Experimental design process as follows Figure 1 As shown.

[0050] (2.1) Construction of a nephrotoxic serum nephritis (NTN) model:

[0051] Five days before the start of the experiment, mice (the model group, positive control group, low-dose psyllium glycoside group and high-dose psyllium glycoside group) were pre-sensitized by intraperitoneal injection of 0.25 mg of anti-glomerular basement membrane IgG. Starting from day 0 of the experiment, 100 μL of nephrotoxic serum (NTS) was injected via tail vein to induce the NTN model. The drug was administered continuously for 7 days starting from day 0.

[0052] (2.2) Experimental grouping and intervention methods:

[0053] Female C57BL / 6 mice were randomly divided into 5 groups of 6 mice each, and were subjected to the following treatments:

[0054] Normal control group (NC): No treatment was given, and an equal volume of DMSO solution was injected intraperitoneally daily starting from day 0 of the experiment;

[0055] Model group (NTN): An NTN model was established, and an equal volume of DMSO solution was injected into the peritoneum daily starting from day 0 of the experiment;

[0056] Positive control group (prednisone): An NTN model was established, and prednisone 5 mg / kg / day was administered by gavage daily starting from day 0 of the experiment;

[0057] Low-dose psyllium phosphate group (His 5 mg / kg): NTN model was established, and psyllium phosphate 5 mg / kg / day was injected intraperitoneally daily starting from day 0 of the experiment;

[0058] High-dose psyllium ether group (His 10 mg / kg): An NTN model was established, and psyllium ether was injected intraperitoneally 10 mg / kg / day starting from day 0 of the experiment.

[0059] (3) Specimen collection:

[0060] On day 7 of the experiment, after fasting but not watering overnight, urine was collected from mice (metabolic cage method, 4 hours). Blood was collected from the orbital cavity after anesthesia, and serum was separated. Mice were euthanized, and both kidneys were removed, weighed, and one kidney was fixed in 4% paraformaldehyde or embedded in OCT for pathological examination. The other kidney was preserved in liquid nitrogen for molecular biological analysis.

[0061] (4) Experimental results:

[0062] (4.1) Behavioral observation:

[0063] During the experiment, the condition of mice in each group was observed throughout. Mice in the normal control group moved freely, had shiny fur, normal diet, and steady weight gain. Mice in the model group began to lose weight on the first day after modeling, and some mice were found to have positive urine protein by urine protein test strips. Afterward, they began to show symptoms such as slow weight gain, lethargy, reduced activity, dull fur, arched back, and curled-up posture. The above symptoms of mice in the psyllium treatment groups (low-dose group and high-dose group) were significantly improved compared with those in the model group. In particular, the mice in the high-dose psyllium group had a good mental state and relatively shiny fur.

[0064] (4.2) Blood and urine test results:

[0065] The urine protein / creatinine ratio (UTP / UCr), blood urea nitrogen (BUN), serum creatinine (Cr), serum albumin (ALB), triglycerides (TG), and cholesterol (CH) were measured using a fully automated biochemical analyzer. The test results are as follows: Figure 2 As shown, P<0.05, P<0.01, P<0.001, P<0.0001.

[0066] Depend on Figure 2The data showed that, compared with the normal control group, the model group mice had significantly increased serum BUN, Cr, TG, and CH, significantly decreased ALB, and significantly increased urinary UTP / UCr, all indicating successful establishment of the animal model. Compared with the model group, psyllium brevicornuate and hormone treatment significantly reduced serum BUN, Cr, and CH, increased ALB, and decreased urinary UTP / UCr in NTN mice (P < 0.05). The therapeutic effect of high-dose psyllium brevicornuate (10 mg / kg) was similar to that of the positive control group (prednisone), suggesting that 10 mg / kg is the optimal dose for psyllium brevicornuate to exert its therapeutic effect.

[0067] In addition, to assess the potential impact of psyllium argentide on liver function in mice with nephrotoxic serum nephritis (NTN), serum liver function indicators were measured. Figure 3 As shown, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in all groups of mice remained within the normal range, and there were no statistically significant differences among the groups (P > 0.05). These results indicate that, within the experimental dosage range, psyllium arvense intervention did not cause hepatocyte damage in NTN mice and had no significant toxic side effects on liver function, suggesting that it has good liver safety.

[0068] (4.3) Renal pathological evaluation (PAS staining):

[0069] PAS staining results of mice in each experimental group are as follows: Figure 4 As shown (scale bar = 50 μm), the top row shows glomerular morphology, and the bottom row shows renal tubules. It can be seen that the kidney structure of mice in the normal control group is normal; mice in the model group show obvious glomerular mesangial proliferation, renal tubular dilation, interstitial edema, and inflammatory infiltration, with a large amount of PAS-positive material deposited in the mesangial matrix; the psyllium ether treatment groups (His 5 mg / kg and 10 mg / kg) dose-dependently reduced glomerular lesions and tubulointerstitial damage, reduced basement membrane thickening, decreased mesangial matrix deposition, and improved renal histopathological changes.

[0070] (4.4) Renal IgG deposition and complement C3 deposition:

[0071] Immunofluorescence staining was used to detect IgG and complement C3 deposition in the kidneys, and the results were as follows: Figure 5 As shown (scale bar = 50 μm), the top row shows IgG deposition, and the bottom row shows complement C3 deposition. It is evident that the model group mice exhibited abundant IgG and C3 deposition in the glomerular region, distributed in granular or clumped patterns in the mesangial area and capillary loops. Treatment with psyllium ether significantly reduced IgG and C3 deposition in the kidneys, suggesting that it may exert a renal protective effect by inhibiting immune complex deposition and complement activation.

[0072] (4.5) Detection of relative mRNA expression level of Angptl4:

[0073] RNA was extracted from mouse kidney tissue and analyzed by qPCR to determine the relative expression level of Angptl4 mRNA in mouse kidney tissue. The test results are as follows: Figure 6 As shown, compared with the normal control group, the expression of Angptl4 in the kidneys of mice in the model group was significantly increased; treatment with high-dose psyllium ether (His 10 mg / kg) significantly reduced the expression of Angptl4.

[0074] This invention explores the pharmacodynamic effects of psyllostigmine in the treatment of lupus nephritis (LN) by constructing an NTN model. The NTN model is based on an immune complex-mediated glomerular injury mechanism, which is highly consistent with the pathogenesis of human lupus nephritis. This model is characterized by glomerular immune complex deposition, complement activation, and proliferative lesions, which are fundamentally different from other kidney injury models such as endotoxin-induced acute inflammatory kidney injury (without immune complexes) and doxorubicin nephropathy (primarily podocyte toxicity). The NTN model is specific and irreplaceable for studying the pathogenesis and drug evaluation of human lupus nephritis.

[0075] This invention has demonstrated the pharmacodynamic effects of psyllium brevicornu in the treatment of lupus nephritis through the above experiments. The results show that the unique pharmacological effects of psyllium brevicornu can be used to reduce kidney damage in lupus nephritis, thereby reducing proteinuria and renal function impairment, and alleviating pathological changes caused by immune complex deposition. At the same time, this invention preliminarily reveals that Angptl4 may be its key target, providing important experimental evidence and theoretical support for the clinical translation of psyllium brevicornu and the pharmacodynamic material basis research of traditional Chinese medicine compound lupus formula.

[0076] The invention relates to psyllium husk, which achieves therapeutic effects similar to those of hormones and can largely meet the urgent clinical needs for targeted therapy and kidney protection in lupus nephritis.

[0077] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0078] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0079] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable way without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. Use of a high pinitol or a pharmaceutically acceptable salt, ester, solvate thereof in the preparation of a medicament for treating lupus nephritis.

2. Use according to claim 1, characterized in that, The medicament further comprises a pharmaceutically acceptable excipient; Preferably, the pharmaceutically acceptable excipient comprises any one or a combination of at least two of a filler, a binder, a wetting agent, a disintegrant, a solubilizer, an osmotic pressure regulator, a coating material, a colorant, a pH regulator, an antioxidant, or a bacteriostatic agent. Preferably, the dosage form of the medicament comprises any one of a dispersible tablet, a sustained-release tablet, an effervescent tablet, an enteric tablet, a sustained-release capsule, an enteric capsule, a granule, an injection, a dripping pill, or a solution.

3. Use of a high pinitol or a pharmaceutically acceptable salt, ester, solvate thereof in the preparation of a modulator of a serum biochemical index related to kidney function.

4. Use according to claim 3, characterized in that, The modulator of a serum biochemical index related to kidney function comprises any one or a combination of at least two of a modulator for reducing urea nitrogen level, a modulator for reducing creatinine level, a modulator for reducing cholesterol level, or a modulator for increasing albumin level.

5. Use of a high pinitol or a pharmaceutically acceptable salt, ester, solvate thereof in the preparation of a medicament for improving histopathological damage of a kidney.

6. Use according to claim 5, characterized in that, The histopathological damage of a kidney comprises any one or a combination of at least two of glomerular lesion, tubulointerstitial damage, basement membrane thickening, or mesangial matrix deposition.

7. Use of a high pinitol or a pharmaceutically acceptable salt, ester, solvate thereof in the preparation of an immune complex deposition inhibitor.

8. Use according to claim 7, characterized in that, The immune complex deposition inhibitor comprises an IgG deposition inhibitor and / or a C3 deposition inhibitor.

9. Use of a high pinitol or a pharmaceutically acceptable salt, ester, solvate thereof in the preparation of an angiopoietin-like protein expression inhibitor.

10. Use according to claim 9, characterized in that, The angiopoietin-like protein expression inhibitor comprises an angiopoietin-like protein 4 expression inhibitor.

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