Use of apigenin and derivatives thereof for the preparation of a medicament for the treatment or prevention of kidney disease

Abstract

The present application relates to the field of pharmaceutical chemistry, and particularly relates to application of apigenin and derivatives thereof or pharmaceutically acceptable salts thereof in preparation of drugs for treating or preventing kidney diseases, for example, chronic renal failure diseases. The research results of the present application show that apigenin can obviously improve symptoms of kidney diseases, especially chronic renal failure diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical chemistry, in particular to the application of apigenin and its derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for treating or preventing kidney diseases, such as chronic renal failure. BACKGROUND

[0002] Apigenin (C 15 H 10 O5, CAS: 520-36-5) belongs to flavonoids, has the carcinogenic activity of inhibiting carcinogens, is an antiviral drug for treating human immunodeficiency virus (HIV) and other viral infections, an inhibitor of mitogen-activated protein kinase (MAP kinase), a treatment for various inflammations, an antioxidant, a sedative, a tranquilizer, and a blood pressure reducer. Compared with other flavonoids (quercetin, kaempferol), it has the characteristics of low toxicity and non-mutagenicity.

[0003]

[0004] In the existing related patents, only the application of apigenin (application number: CN201710857425.8) is found. The content of this patent is that apigenin can inhibit the activity of Dipeptidyl peptidase IV (DPP-IV) and / or pharmaceutically acceptable solvates. Such compounds can be used to treat diabetes, such as type 2 diabetes, hyperglycemia, metabolic syndrome, hyperinsulinemia, obesity, cardiovascular diseases and abnormalities such as atherosclerosis, cerebrovascular diseases, central nervous system diseases or abnormalities including schizophrenia, anxiety, bipolar depression, depression, insomnia, cognitive impairment, gastrointestinal diseases and abnormalities, cancer, inflammation and inflammatory diseases, respiratory diseases and abnormalities, skeletal muscle abnormalities, osteoporosis, menopausal symptoms or abnormalities, periodontal diseases such as gingivitis, and various immune-regulating diseases. The application of apigenin in inhibiting the activation of Wnt / β-catenin signaling pathway in colon cancer cells (application number: CN201710663618.X) is disclosed.

[0005] Chronic renal failure is a group of clinical symptoms composed of chronic kidney disease developing to the late stage caused by various causes. According to the degree of renal function damage, chronic renal failure is divided into 4 stages: decreased renal reserve function, patients have no symptoms; compensatory period of renal insufficiency; decompensated renal function (azotemia period), patients have fatigue, loss of appetite and anemia; uremia stage, with uremia symptoms.

[0006] Apigenin has been reported to have the effects of treating diabetes and preventing colon cancer. However, the prior art has not disclosed and suggested the use of apigenin for treating or preventing chronic kidney disease, especially chronic renal failure. SUMMARY

[0007] An object of the present application is to provide a new use of apigenin and derivatives thereof or pharmaceutically acceptable salts thereof in the field of pharmaceutical applications, i.e. the use of apigenin and derivatives thereof or pharmaceutically acceptable salts thereof in the preparation of a medicament for treating or preventing kidney disease, for example, chronic renal failure disease.

[0008] The present application provides the use of apigenin and derivatives thereof or pharmaceutically acceptable salts thereof in the preparation of a medicament for treating or preventing kidney disease, for example, chronic renal failure disease.

[0009] The present application provides a pharmaceutical composition for treating or preventing kidney disease, for example, chronic renal failure, comprising a therapeutically effective amount of apigenin and derivatives thereof or pharmaceutically acceptable salts thereof as an active ingredient. The pharmaceutical composition can further comprise a pharmaceutically acceptable carrier.

[0010] The apigenin and derivatives thereof or pharmaceutically acceptable salts thereof according to the present application can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents. The therapeutic agents include, but are not limited to, angiotensin II AT1 receptor antagonists, such as candesartan cilexitil, eprosartan, irbesartan, losartan potassium, telmisartan; converting enzyme inhibitors, such as alacepril, benazepril, captopril, cilazapril, enalapril, enalaprilat; diuretics, such as hydrochlorothiazide, bendroflumethiazide, chlorthalidone.

[0011] "Kidney disease" refers to diabetic nephropathy, chronic renal failure, chronic nephritis, end-stage kidney disease, kidney hypertrophy, kidney hyperplasia, glomerular sclerosis, or glomerulonephritis.

[0012] "Pharmaceutically acceptable salt" refers to a salt formed with inorganic acids such as phosphoric acid, sulfuric acid, hydrochloric acid, or organic acids such as acetic acid, tartaric acid, citric acid, malic acid, or acidic amino acids such as aspartic acid and glutamic acid, or a salt formed after esterification or amidation with the above acids and inorganic bases such as sodium, potassium, calcium, aluminum, and ammonium.

[0013] "pharmaceutically acceptable carrier" means one or more compatible solid or liquid filler or gel materials, which are suitable for human use and must have sufficient purity and low toxicity. "compatible" means that the composition components are capable of being commingled with the active ingredients of the present application and with each other in the pharmaceutical composition with no interaction that would substantially reduce the pharmaceutical efficacy of the active ingredients. Examples of suitable pharmaceutically acceptable carriers are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyhydric alcohols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., lecithin), wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0014] The pharmaceutical composition for treating or preventing kidney disease, e.g., chronic renal failure, according to the present application can be prepared in any of the various dosage forms of pharmaceuticals. For example, it can be in the form of oral or injectable, such as capsules, tablets, dispersible tablets, infusion solutions, injections, lyophilized preparations, oral preparations, suspensions, pills, emulsions, powders, or gels. The pharmaceutical composition for treating or preventing kidney disease, e.g., chronic renal failure, according to the present application can be prepared into any desired dosage form using techniques commonly used in the art.

[0015] The content of this patent application is the pharmacological study of apigenin and its derivatives or pharmaceutically acceptable salts thereof on kidney disease, e.g., chronic renal failure, and the test results show that they can effectively improve the symptoms of chronic renal failure.

[0016] Advantages

[0017] 1. The pharmacological study of apigenin and its derivatives or pharmaceutically acceptable salts thereof on kidney disease, especially chronic renal failure, shows that apigenin can significantly improve the symptoms of chronic renal failure.

[0018] 2. The raw material of apigenin and its derivatives or pharmaceutically acceptable salts thereof and different types of dosage forms thereof are effective for kidney disease, especially chronic renal failure. DETAILED DESCRIPTION

[0019] The present application will be further described below in conjunction with specific examples, but the present application is not limited thereto.

[0020] Example 1

[0021] 1) Materials:

[0022] ​Animals: 30 outbred SD rats, male, 300g±20g, provided by Shanghai Slac Laboratory Animal Co. Ltd, license number: SCXK (Shanghai) 2003-0003, fed with commercial solid feed for 7 days to adapt to the environment.

[0023] 2) Drugs:

[0024] Dexamethasone sodium phosphate injection, produced by Shanghai General Pharmaceutical Co., Ltd., content: 5mg / mL, batch number: 060608;

[0025] Apigenin was purchased from Jiangsu Jiangshan Pharmaceutical Co., Ltd., batch number: 061010. The required different concentrations of drug solution were prepared with solvent 0.5wt.% DMSO.

[0026] 3) Method: SD rats were randomly divided into five groups according to body weight, namely sham operation group, model group, positive drug group (dexamethasone) and low-dose apigenin group and high-dose apigenin group, 6 rats in each group. The 5 / 6 nephrectomy chronic renal failure (CRF) model of rats was used. The SD rats were anesthetized by intraperitoneal injection of 2wt.% pentobarbital (30mg / kg wt.), the skin was disinfected, and the left kidney was fully exposed through a left dorsal or abdominal incision. The kidney capsule was stripped, and about 1 / 3 of the upper and lower poles was quickly removed. Gelatin sponge was used to press the bleeding, and a few drops of thrombin solution were added to speed up the blood clotting. After there was no active bleeding, the layers were sutured.

[0027] The second operation modeling was performed 7 days after the operation. The right kidney was exposed by incision on the right back, the renal pedicle was ligated, the right kidney was removed, and the weight of the removed right kidney tissue was measured.

[0028] Two days after the second operation modeling, the positive drug group was given intramuscular injection of dexamethasone (0.1mg / kg wt.); the low-dose apigenin group and the high-dose apigenin group were given apigenin 4μM / kg wt. and 16μM / kg wt. by gavage, respectively, and the gavage volume was 0.1mL / kg wt.; a sham operation group was also set up, and the operation process was the same as the other groups except that the kidney was retained. The sham operation group and the model group were given the same volume of solution containing 0.1wt.% DMSO and 5wt.% glucose injection (referred to as DMSO: sugar water). The different treatment methods were given to the SD rats in each group for 34 days according to the method shown in Table 1, and all the animals were free to take feed and water.

[0029] On the second day after the second operation modeling, all rats received intramuscular injection of tetracycline (as a fluorescent dye) at 25mg / kg wt.; on the 30th day, intramuscular injection of calcein was performed at 15mg / kg wt.

[0030] Table 1 Treatment method of animals in each group

[0031]

[0032] 4) Each day, the test animals were observed routinely, and 24h urine was collected for urine protein detection every week. After the end of administration, bone density was tested, blood was taken from the abdominal aorta, serum was separated, and hemogram and biochemical indexes were detected. The animals were dissected, the kidneys were taken, weighed, and opened and organ coefficients were calculated, then fixed in 10% formalin solution, routinely embedded, sectioned, stained, and subjected to pathological examination.

[0033] The specific detection indexes and methods are as follows:

[0034] (1) General observation: every day;

[0035] (2) Body weight, once a week, and survival rate were calculated;

[0036] (3) Biochemical examination: TOCHE COBAS MIRA full-automatic biochemical analyzer produced in Switzerland was used to detect blood creatinine, serum total protein and albumin, serum urea nitrogen, K + , Na + , Ga 2+ , P, Mg 2+ and b-ALP, Zenyth200st enzyme label instrument produced in Austria and rat parathyroid hormone enzyme-linked immunoassay (PTH) ELISA kit (product number: E0465r) were used to detect the content of serum PTH (parathyroid hormone), and the examination was performed after drug withdrawal;

[0037] (4) 24h urine volume and urine component analysis: AUTION JET urine ten-item analyzer produced in Japan was used to detect urine protein content, once a week;

[0038] (5) Hematology examination: AB&VET ABC (169) blood cell counter produced in France was used to detect red blood cell count, hemoglobin and hematocrit, and the examination was performed after drug withdrawal;

[0039] (6) Bone density examination: the examination was performed after drug withdrawal, bone growth rate was observed, after the tibia and fibula were removed from soft tissues, the tibia was retained in 70% alcohol, refrigerated for two days, then transferred to 70% alcohol, refrigerated, and the total bone density, trabecular bone density and cortical bone density were detected by peripheral quantitative computed tomography (pQCT, peripheral Quantitative Computed Tomography, XCTReasearch SA) produced by Germany Stratec company. The fibula was immediately frozen with dry ice or liquid nitrogen, then wrapped with gauze soaked with normal saline, and stored in a refrigerator below-70 degrees for future frozen section and enzyme analysis;

[0040] (7) Gross histological observation, microscopic pathological histological examination of kidney.

[0041] ① Renal pathological morphology observation: gross and histological observation, calculation of kidney coefficient, PAS special staining, and calculation of mesangial / glomerular area ratio by IMS cell image analysis system and medical image analysis software developed by Shanghai Shenteng Information Technology Co., Ltd.

[0042] ② Proliferation cell nuclear antigen (PCNA) immunostaining of paraffin sections was performed using PCNA immunohistochemical staining kit (product number: SA2024) provided by Wuhan Doctorde Biological Engineering Co., Ltd. to calculate the proportion of positive cell nuclei in glomerulus.

[0043] 5) Data processing:

[0044] Results were expressed as , and significance test was performed by analysis of variance.

[0045] 6) Results

[0046] Table 2, effect of each test substance on body weight of rats g)

[0047]

[0048] Note: compared with the sham operation group # p<0.05, ## p<0.01; compared with the model group *p<0.05, **p<0.01.

[0049] From the results in Table 2, it can be seen that compared with the model group, the high dose and low dose groups of apigenin had significant differences in average body weight of animals (P<0.05).

[0050] Table 3, effect of each test substance on survival rate of rats (%)

[0051]

[0052] The sham operation group animals showed alertness, fast reaction, dense and neat fur, normal diet, and no animal death. The other groups of animals were in poor spirits, slow movement, loose and withered fur, loss of luster, loss of appetite, among which the model group was the most obvious. There were deaths of model animals every week, and the death conditions are shown in Table 3.

[0053] Table 4, effect of each test substance on urinary protein of rats mg)

[0054]

[0055]

[0056] Note: Compared with the sham operation group # p<0.05, ## p<0.01; compared with the model group *p<0.05, **p<0.01.

[0057] From the results of Table 4, it can be seen that the urine protein content of the high-dose and low-dose groups of apigenin animals had significant differences (P<0.05) compared with the model group.

[0058] Table 5, Effect of each test substance on blood and biochemical indicators of rats

[0059]

[0060] Note: Compared with the sham operation group # p<0.05, ## p<0.01; compared with the model group *p<0.05, **p<0.01.

[0061] Table 6, Effect of each test substance on blood and biochemical indicators of rats

[0062]

[0063] Note: Compared with the sham operation group # p<0.05, ## p<0.01; compared with the model group *p<0.05, **p<0.01.

[0064] From the results of Tables 5 and 6, it can be seen that there were no significant differences (P>0.05) in blood and biochemical indicators of animals in each group compared with the model group.

[0065] Table 7, Effect of each test substance on bone density of rats mg / cm 3 )

[0066]

[0067]

[0068] Note: Compared with the sham operation group # p<0.05, ## p<0.01; compared with the model group *p<0.05, **p<0.01.

[0069] Compared with the model group, the trabecular bone density of the high-dose and low-dose groups of apigenin animals had significant differences (P<0.05).

[0070] Table 8, the influence of each test object on the indicators related to kidney pathology of rats

[0071]

[0072] Note: compared with the sham operation group # p<0.05, ## p<0.01; compared with the model group, *p<0.05, **p<0.01.

[0073] Compared with the model group, the mesangial glomerular area ratio of the high-dose and low-dose groups of apigenin had significant differences (P<0.05); compared with the model group, the high-dose group of apigenin had significant differences in the proportion of proliferative cell nuclei in the kidney (P<0.05).

[0074] Taking the model group as the control, the degree of kidney injury, the high-dose and low-dose groups of apigenin had less kidney injury, see Table 8 for details.

[0075] The model group was successfully modeled, the positive drug dexamethasone had a certain relieving effect on the symptoms of chronic renal failure, and 16 μM / kg of apigenin had a certain improving effect on the symptoms of chronic renal failure.

Claims

1. The use of apigenin or its pharmaceutically acceptable salts in the preparation of medicaments for the treatment of kidney disease; The kidney disease mentioned is chronic renal failure.

2. Use of the pharmaceutical composition in the preparation of a medicament for treating kidney disease; The pharmaceutical composition contains a therapeutically effective amount of apigenin or a pharmaceutically acceptable salt thereof as the active ingredient, and optionally a pharmaceutically acceptable carrier; Alternatively, the pharmaceutical composition comprises a therapeutically effective amount of apigenin or a pharmaceutically acceptable salt thereof as an active ingredient, along with other pharmaceutically acceptable therapeutic agents, and optionally a pharmaceutically acceptable carrier; The kidney disease mentioned is chronic renal failure.

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