阅读说明：本技术 一种具有肾脏保护作用的药物组合物 (Pharmaceutical composition with kidney protection effect ) 是由 段洪伟 白洁 张磊 陈光亮 徐希平 于 2019-08-28 设计创作，主要内容包括：本发明涉及一种具有肾脏保护作用的药物组合物。该药物组合物由一种内皮素抑制剂、依那普利、叶酸类物质及药剂学上可接受的载体组成,内皮素抑制剂包括阿曲生坦。本发明提供了该药物组合物在制备用于预防、治疗或延缓慢性肾脏疾病和糖尿病性肾病的药物中的用途。通过本发明的实施,提供给患者该药物组合物,可以提高疗效,增加治疗依从性,降低医药费用。(The invention relates to a pharmaceutical composition with kidney protection effect. The medicine composition consists of an endothelin inhibitor, enalapril, folic acid substances and a carrier acceptable in pharmaceutics, wherein the endothelin inhibitor comprises atrasentan. The invention provides application of the pharmaceutical composition in preparing a medicament for preventing, treating or delaying chronic kidney diseases and diabetic nephropathy. Through the implementation of the invention, the pharmaceutical composition is provided for patients, the curative effect can be improved, the treatment compliance is increased, and the medical cost is reduced.)

1. A pharmaceutical composition comprising:

(1) a pharmaceutically acceptable dose of an endothelin inhibitor;

(2) a pharmaceutically acceptable dose of enalapril;

(3) a pharmaceutically acceptable dose of folic acid;

(4) a pharmaceutically acceptable carrier.

2. The pharmaceutical composition of claim 1, wherein: the endothelin inhibitor is atrasentan, and the medicinal dosage is 0.375-1.5 mg.

3. The pharmaceutical composition of claim 1, wherein: the medicinal dosage of the enalapril is 5-20 mg, preferably 5-10 mg.

4. The pharmaceutical composition of claim 1, wherein: the folic acid substances comprise folic acid, 5-methyltetrahydrofolic acid, leucovorin, folinic acid, calcium levofolinate, folic acid medicinal salt and the like, and the medicinal dosage is 0.4-1.6 mg, preferably 0.4-0.8 mg.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the composition is in the form of an oral preparation.

6. Use of the pharmaceutical composition of any one of claims 1-5 in the preparation of a medicament for preventing, treating or delaying chronic kidney disease.

7. Use of the pharmaceutical composition of any one of claims 1-5 in the preparation of a medicament for preventing, treating or delaying diabetic nephropathy.

8. Use of a pharmaceutical composition according to any one of claims 1 to 5 in the manufacture of a medicament for the treatment of hypertension associated with impaired renal function.

Technical Field

The invention relates to a pharmaceutical composition for combined application of an endothelin inhibitor, enalapril and folic acid substances and application thereof, belonging to the field of pharmacy.

Background

Chronic Kidney Disease (CKD) is a Chronic kidney structure and function disorder caused by various causes, and it is estimated that about 5 million people worldwide have kidney lesions of different degrees, so CKD has become a global health problem. The medical service cost of the CKD patient is 1.8 times of that of the non-chronic kidney disease crowd, and the average medical service cost of the dialysis patient is 10.3 times of that of the non-chronic kidney disease crowd, so that huge economic burden is brought to the patient, and the life quality of the patient is seriously influenced. In addition, the incidence and the fatality rate of cardiovascular diseases of CKD patients are obviously higher than those of common people, and the disease is early and fast in progress, which is the leading cause of death and disability of chronic kidney diseases.

Diabetic nephropathy (DKD) refers to glomerulosclerosis caused by Diabetic microangiopathy, is one of the most common chronic complications of microvascular diabetes, and is also one of the major causes of cardiovascular events and death in Diabetic patients. According to epidemiological statistical studies, it is predicted that by 2030, worldwide diabetic patients will reach 3.66 million, while diabetic nephropathy patients will exceed 1 million. In China, diabetic nephropathy has become the second leading cause of end-stage renal failure, second only to glomerulonephritis. .

Studies have shown that folate supplementation can reduce the risk of stroke by 18% overall. The layering analysis shows that when the traditional Chinese medicine composition is used for primary prevention, the stroke risk is reduced by 25%; when applied to people who do not receive foods and add folic acid, the risk of cerebral apoplexy is reduced by 25% [ Wang X, Qin X, Demirtas H, et al.effectiveness of Folic acid supplementation in stroke preservation a meta-analysis.Lancet 2007; 369: 1876-; additionally, enalapril folate Therapy compares to enalapril alone [ Xin Xu, Xianhui qin.et al.efficacy of Folic Acid Therapy on the progression of Chronic Kidney disease. TAMA Inter Med.2016; 176: 1443-. Significantly delays CKD progression in mild to moderate CKD patients. Long-term folic acid supplementation can also significantly reduce the intimal-media thickness progression, with better efficacy in chronic kidney disease people or high risk people with cardiovascular disease [ Qin X, Xu M, Zhang Y, et al. Effect of folic acid administration on the progression of myocardial intervention-media diseases, A meta-analysis of random controlled variants. Atherosclerosis, 2012; 222: 307-.

Endothelin (ET) is derived from endothelial cells and has potent and durable vasoconstrictive activity. Pathophysiological studies have shown that ET-1 is mediated by two subtypes, endothelin receptors ETA and ETB, in various tissues and is involved in the physiological and pathological processes of various cardiovascular diseases. Both ETA and ETB promote vasoconstriction in smooth muscle cells, whereas ETB receptors in endothelial cells can clear ET-1, which plays a role in vascular protection by regulating nitric oxide coupling, vasodilation and anti-cell proliferation. The research shows that [ Weizhen, ET al ] the novel angiotensin II receptor and endothelin receptor dual antagonist research progresses pharmaceutical progress 2006,30(5):193-199 ], endothelin has angiotensin (Ang) converting enzyme-like activity and can promote the conversion of Ang I to Ang II, and the application of ET receptor antagonist can reduce Ang II, while Ang II can promote the expression and release of ET, the mechanism may be that a regionalized ET-Ang II interaction system exists in the cardiovascular system, and the mechanism participates in the regulation of local circulation homeostasis [ Laplan M A, ET al.

Atrasentan is an ETA receptor antagonist with high efficiency and high selectivity (the affinity to ETA is 1860 times that of ETB), and can reverse or block the promotion effect of ET-1 on cell proliferation, angiogenesis and bone remodeling and the influence thereof on hemodynamics. Atrasentan has been shown to reduce proteinuria levels in diabetic nephropathy and chronic kidney disease patients and as a selective endothelin receptor antagonist, fewer adverse events during treatment.

Disclosure of Invention

The invention aims to provide a pharmaceutical composition with remarkable curative effect on chronic kidney diseases or diabetic nephropathy.

In order to achieve the purpose, the invention adopts the following technical scheme:

a pharmaceutical composition comprising:

(1) a pharmaceutically acceptable dose of an endothelin inhibitor;

(2) a pharmaceutically acceptable dose of enalapril;

(3) a pharmaceutically acceptable dose of folic acid;

(4) a pharmaceutically acceptable carrier.

In the pharmaceutical composition provided by the invention, the endothelin inhibitor mainly refers to atrasentan, and the atrasentan can exist in the form of salts, lipids, active metabolites or medicinal precursors. The atrasentan is used as a main medicinal component, the salt, lipid, active metabolite or medicinal precursor form of the atrasentan is also in the protection range of the application, the medicinal dose of the atrasentan is 0.375-1.5 mg, and the medicinal dose of the lipid, active metabolite or medicinal precursor form of the atrasentan can be correspondingly converted.

In the pharmaceutical compositions provided herein, enalapril may be present in the form of salts, lipids, active metabolites, or pharmaceutically acceptable precursors. The enalapril provided by the invention is used as a medicinal component, the existing forms of salts, esters, active metabolites, medicinal precursors and the like of the enalapril are also within the protection scope of the application, the medicinal dose of the enalapril is 2.5-10 mg, and the medicinal doses of various forms can be mutually converted.

The folic acid substances in the pharmaceutical composition provided by the invention comprise folic acid, leucovorin calcium, 5-methyltetrahydrofolic acid (L-methylfolic acid), folate, active metabolites of folic acid or folate and substances capable of releasing/generating folic acid in vivo. The pharmaceutical dosage of folic acid compounds is 0.4-1.6 mg, preferably 0.4-0.8 mg, and the pharmaceutical dosages of various forms thereof can be converted to each other.

In the present invention, the pharmaceutically acceptable dose of the active ingredient of the composition means a dose range in which the active ingredient of the composition exerts its pharmacological effect when combined with other active ingredients in the composition. The dosage range of the effective components of the composition is also the optimal dosage range, so that the patients can obtain more benefits.

Through research, the pharmaceutical composition of the invention:

preferably, the pharmaceutical composition provided by the invention comprises 0.75mg of asiracetam, 10mg of enalapril and 0.4mg of folic acid.

As another preferred mode, the pharmaceutical composition provided by the invention comprises 0.75mg of asistantan, 10mg of enalapril and 0.8mg of folic acid.

As another preferred mode, the pharmaceutical composition provided by the invention comprises 1mg of asiracetam, 10mg of enalapril and 0.4mg of folic acid.

As another preferred mode, the pharmaceutical composition provided by the invention comprises 1mg of asiracetam, 10mg of enalapril and 0.8mg of folic acid.

The pharmaceutical composition also comprises pharmaceutically acceptable carriers, can be prepared into common oral preparations, including common tablets, common capsules, granules and the like, and the pharmaceutically acceptable carriers comprise excipients which are helpful for preparing active compounds into the pharmaceutical preparations when the pharmaceutical composition is prepared into the tablets, such as one or a combination of more of pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, lactose, magnesium stearate, polyvinylpyrrolidone and the like, and belong to the common knowledge in the field.

The pharmaceutical compositions of the present invention may alternatively be used in a "combination kit". The above-mentioned "combined medicine box" is a box-shaped container, in which the medicine combination with several dosage forms is placed, and its administration instruction. The combined medicine box is more suitable for individual medicine.

The invention has the beneficial effects that: the pharmaceutical composition is suitable for CKD and diabetic nephropathy, and can significantly improve renal function of patients. The active ingredients of the pharmaceutical composition provided by the invention are atrasentan, enalapril and folic acid substances, wherein the atrasentan is a high-selectivity endothelin inhibitor, can reverse or block the promotion effect of ET-1 on cell proliferation, angiogenesis and bone remodeling and the influence thereof on hemodynamics, and reduces the protein urine level of patients with diabetic nephropathy and chronic nephropathy. Enalapril is an angiotensin converting enzyme inhibitor, can effectively protect the kidney when being used together with folic acid, can also reduce the risk of cardiovascular and cerebrovascular diseases, can synergistically improve the kidney function when being used together, and has the function of reducing blood pressure for patients with hypertension, so that the pharmaceutical composition provided by the invention is more suitable for treating the patients with the hypertension accompanied with renal function damage. The synergistic improvement effect of the pharmaceutical composition provided by the invention cannot be obvious by single medicine or two-two combination. The pharmaceutical composition provided by the invention also has the characteristics of low dosage and less side effect. In addition, the invention can also facilitate the patients to take medicine, improve the compliance of the patients, reduce the medical cost and have better market prospect.

The invention will now be further described with reference to specific embodiments, it being understood that the dosages selected in the examples are not intended to be limiting of the invention, and that any equivalent replacement in the art which has been made in accordance with the teachings of the present invention is deemed to be within the scope of the present invention.

Detailed Description

Examples 1-6 atrasentan + enalapril + folic acid tablets (1000 tablets amount)

The preparation process comprises the following steps:

mixing atrasentan, enalapril and folic acid, adding carboxymethyl starch sodium and sodium dodecyl sulfate, mixing, adding microcrystalline cellulose and pregelatinized starch, mixing, adding appropriate amount of 10% polyvidone ethanol solution to obtain soft material, granulating, drying, grading, mixing with appropriate amount of magnesium stearate and granule with water content of about 3%, and tabletting to obtain 1000 tablets.

Examples 7-12 atrasentan + enalapril + folic acid capsules (1000 capsules)

The preparation process comprises the following steps: pulverizing the raw materials and adjuvants, sieving with 80 mesh sieve, and drying. Taking atrasentan, enalapril and folic acid, uniformly mixing, adding lactose crystal, polyvinylpyrrolidone, microcrystalline cellulose and sodium carboxymethyl starch, uniformly mixing, preparing a soft material by using superfine silica gel powder, granulating by using a 20-mesh sieve, drying at 60 ℃ for about 2 hours, finishing granules by using the 20-mesh sieve, controlling the water content of the granules to be 2-3%, uniformly mixing the dried granules with a proper amount of 1% magnesium stearate, detecting a semi-finished product, measuring the content, and filling into hollow capsules to obtain 1000 capsules. Care should be taken to avoid light during the preparation process. And packaging the aluminum-plastic bubble cap after the finished product is qualified by inspection, and storing in dark place.

Example 13: the composition of atrasentan and enalapril folic acid has kidney protection effect on chronic nephrotic rats.

Firstly, molding:

adult Wistar rats with the weight of 200-220g are male, animals with the weight deviation of more than 10% are removed after being fed with common feed and quarantined for 7 days, 10 rats are randomly selected as a normal control group, and the animals are continuously fed with the common feed until the experiment is finished. The animals are fasted for 12 hours before the model building, and are injected by single tail vein of adriamycin (7.9mg/kg), and normal diet and drinking water are recovered after the model building is carried out for 2 hours. And after 4w, detecting the levels of 24h urine protein of all animals, and selecting the molded animals with the 24h urine protein more than or equal to 20mg for subsequent experiments.

(II) grouping and administration:

10 rats in a normal control group are divided into 7 groups randomly, a model group, an atrasentan group of 0.15mg/kg, an enalapril folic acid (efl for short) group of 1.08mg/kg, atrasentan + efl (0.05+0.54) mg/kg, atrasentan + efl (0.075+1.08) mg/kg, atrasentan + efl (0.1+1.08) mg/kg, and atrasentan + efl (0.15+2.16) mg/kg. The animals in each group were gavaged to a volume of 10 ml/kg. The biochemical index is detected after the administration is carried out once a day for 12 weeks.

[ notes ] in the description of the dose, 1.08mg/kg of enalapril per leaf means 1mg/kg of enalapril and 0.08mg/kg of folic acid, 0.54mg/kg of enalapril means 0.5mg/kg of enalapril and 0.04mg/kg of folic acid, and 2.16mg/kg of enalapril means 2mg/kg of enalapril and 0.16mg/kg of folic acid.

(III) detection indexes:

1. urine index: rats were transferred to metabolism cages for 12 weeks after administration, and urine was collected for 24 hours and recorded. Uniformly mixing the collected 24-hour urine, taking the urine, centrifuging at 3000r/min for 10 minutes, taking the supernatant, and measuring the urine protein concentration by a Coomassie brilliant blue method; multiplying by the animal's 24h urine volume to calculate the 24 hour urine protein volume.

2. Blood index: after the administration, animals are anesthetized by pentobarbital sodium, then the abdominal aorta is subjected to blood sampling, a blood sample is kept stand for 2-4h, centrifugation is carried out at 3000r/min for 10 minutes, a supernatant is taken, and the blood creatinine and blood urea nitrogen levels are measured by a biochemical analyzer.

(IV) experimental results:

TABLE 1 Effect of atrasentan + Egyptian on Biochemical indices in Chronic renal disease rats ((n＝10-12)

Note: in comparison with the normal group,^*P<0.05,^**P<0.01; in comparison with the set of models,^#P<0.05，^##P<0.01; compared with the atrasentan group,^△P<0.05,^△△P<0.01. compared with the leaf group,^▲P<0.05,^▲▲P<0.01.

after adriamycin is given for molding, the weight of the animals in the molding group is reduced, the activity is reduced, the stress response caused by adriamycin molding is inferred, the weight begins to stably increase after 3 weeks of injection, the weight of the animals in the normal control group keeps increasing all the time in the experimental period, and the average value of the weight of the animals in the model group is lower than that of the animals in the normal group, but no significant difference exists. No mortality or other abnormalities were found in all groups of animals throughout the experimental period.

The animals are continuously dosed for 12 weeks, and compared with the normal group of rats, the model group of rats have increased blood creatinine, increased BUN and increased urine protein, which indicates that the rats have the symptoms of chronic kidney disease. After administration treatment, the blood creatinine and urine protein of each administration group are obviously reduced and reach obvious difference; however, the single treatment of atrasentan or eletal cannot achieve a very good therapeutic effect. It was unexpectedly found from the data in the above table that atrasentan + efruit exhibited significant synergy at both (0.075+1.08) and (0.1+1.08) mg/kg dosage levels, and further increasing the dosage did not show better efficacy, but the corresponding side effects increased, compared to the single drug.

Example 14: the composition of atrasentan and enalapril folic acid has kidney protection effect on diabetic nephropathy rats.

Firstly, molding:

adult Wistar rats with the weight of 200-220g are male, animals with the weight deviation of more than 10% are removed after 7 days of quarantine by ordinary daily ration feed, 12 rats are randomly selected as a sham operation control group, and the animals are continuously fed by the ordinary feed until the experiment is finished. Feeding high-fat high-sugar feed to the model building animals for 3 weeks, fasting all animals for 12 hours without water supply, performing left kidney ligation operation after pentobarbital sodium anesthesia, only opening the abdominal cavity of the animals in a sham operation group, finding the operation of the left kidney, not ligating, and administering 10 ten thousand units of penicillin to prevent infection within 3 days after the operation. 4w animals recovered after surgery. All animals are fasted for 12 hours without water prohibition, an abdominal cavity one-time injection STZ solution of 35mg/kg is adopted for carrying out a molding operation, the blood sugar of all animals is detected at 3d, 7d and 14d after injection respectively, the urinary protein level of the animals is measured at 24h after administration at 14d, and molded animals with fasting blood sugar (FBG) being more than or equal to 16.7mmol/L and the urinary protein being more than or equal to 20mg after 24h are selected for entering a subsequent test. The molded animals were fed with a high-fat and high-sugar diet throughout the experiment until the end of the experiment.

(II) grouping and administration:

the sham operated control group rats were kept for 12 animals, and animals successfully molded were randomly divided into 6 groups, a model group, an atrasentan 0.1mg/kg group, an enalapril folic acid (abbreviated as efletin) 1.04mg/kg group, atrasentan + efletin 0.075+1.04mg/kg group, atrasentan + efletin 0.1+1.04mg/kg group, and atrasentan + efletin 0.15+2.08mg/kg group. 14 animals per group were gavaged at a volume of 10 ml/kg. The biochemical index is detected after the administration is carried out once a day for 12 weeks.

(III) detection indexes:

1. rat fasting blood sugar

2. Urine index: rats were transferred to metabolism cages for 12 weeks after administration, and urine was collected for 24 hours and recorded. Uniformly mixing the collected 24-hour urine, taking the urine, centrifuging at 3000r/min for 10 minutes, taking the supernatant, and measuring the urine protein concentration by a Coomassie brilliant blue method; and multiplying the urine volume of the animal for 24h to calculate the 24h urine protein volume.

3. Blood index: after the administration, animals are anesthetized by pentobarbital sodium, then the abdominal aorta is subjected to blood sampling, a blood sample is kept stand for 2-4h, centrifugation is carried out at 3000r/min for 10 minutes, a supernatant is taken, and the blood creatinine and blood urea nitrogen levels are measured by a biochemical analyzer.

(IV) experimental results:

the model animals gained weight faster than the control rats during high fat diet feeding, but there was no significant difference between the mean values of the body weights of the two groups. The model building animal begins to have three or more symptoms after STZ model building and surgical ligation of kidney: the weight growth speed of the model animal is reduced due to the toxicity of STZ and the stress injury of the operation, and the 'three more' condition of the model animal is obviously improved after the administration treatment. At the end of the experiment, the average body weight of the model and administration group animals is lower than that of the control group animals, but no significant difference exists. Throughout the experiment, one rat was found dead after 1 week of treatment with the drug, and after dissection, it was found that the ligated kidney tissue was atrophied and hardened, and no obvious lesions were found in other tissues, and no death and abnormalities were found in the remaining animals.

After the continuous administration for 12 weeks, compared with the rats in the sham operation group, the rats in the model group have the advantages of increased blood sugar, increased drinking water amount and increased urine volume, which indicates that the rats have obvious symptoms of diabetes; the increase of blood creatinine, BUN and urine protein of the model group rats shows that the rats have obvious pathological characteristics of diabetic nephropathy. After administration treatment, the urine volume of rats of each administration group is reduced, the blood sugar is not obviously changed, the blood creatinine is obviously reduced, and the urine protein is obviously reduced, which all achieve obvious difference. It was unexpectedly found from the data in table 2 that atrasentan + efolium exhibited significant synergy at both (0.075+1.04) and (0.1+1.04) mg/kg dosage levels, and further increasing the dosage did not show better efficacy, but the corresponding side effects increased, compared to the single drug.

TABLE 2 Effect of atrasentan + Egyptian on Biochemical indices in diabetic nephropathy rats ((n＝12-14)

Note: compared with the group of the pseudo-operation,^*P<0.05,^**P<0.01; in comparison with the set of models,^#P<0.05，^##P<0.01; compared with the atrasentan group,^△P<0.05,^△△P<0.01. compared with the leaf group,^▲P<0.05,^▲▲P<0.01；

as can be seen from examples 13-14, both atrasentan and efylline can obviously improve the renal function of chronic kidney disease rats or diabetic kidney disease rats, but the renal functions cannot reach a better level when being used alone, so that the combination of atrasentan and efylline is selected, and the combination of atrasentan and efylline is unexpectedly found in experiments, and the renal functions are obviously improved compared with the renal functions of single drugs only under the dosage levels of (0.075+ 1.04-1.08) and (0.1+ 1.04-1.08) mg/kg. Clinically, chronic kidney disease patients have edema symptoms of different degrees, and when the dose of atrasentan is too high, the edema of the chronic kidney disease patients is aggravated to cause drug withdrawal (not the serious side effect caused by atrasentan per se).