Combined medicine, application of combined medicine in preparation of medicine for preventing or treating microcirculation disturbance diseases and medicine composition of combined medicine

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a combined medicine, application of the combined medicine in preparation of medicines for preventing or treating microcirculation disturbance diseases, and a medicine composition of the combined medicine. According to the invention, by screening the ratio of the vasodilatation drug to the lucid ganoderma, the influence on microcirculation disturbance is investigated, and the optimal ratio combination for improving the microcirculation function is optimized. Experiments discover that the vasodilatation drug and the lucid ganoderma are compounded, so that the microcirculation function of mice with microcirculation disturbance can be remarkably improved, and particularly, when the dosage ratio of the vasodilatation drug to the lucid ganoderma is (0.65-1.3): 2000, the vasodilatation drug and the lucid ganoderma have a synergistic improvement effect. More choices are provided for preventing or treating diseases related to microcirculation disturbance, and good application prospects are achieved.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a combined drug and its use in preparing a drug for preventing or treating microcirculatory disorders, and a pharmaceutical composition thereof. Background Art

[0002] Microcirculatory disorder refers to a pathological state in which the structure or function of the microvascular system is abnormal, leading to insufficient tissue perfusion or abnormal metabolism. Microcirculatory disorder is closely related to many diseases, especially in the cardiovascular system. For example, coronary microcirculatory disorder (CMD) is a common problem in cardiovascular diseases and is widely present in diseases such as coronary heart disease and acute myocardial infarction. About 40% of patients with coronary heart disease have coronary microvascular dysfunction. Even if coronary angiography does not show obvious stenosis, patients may still experience angina pectoris. In addition, after percutaneous coronary intervention (PCI) for patients with acute myocardial infarction, although the large blood vessels are recanalized, more than half of the patients' distal coronary microcirculation is not effectively restored, resulting in continued angina pectoris. In addition to cardiovascular diseases, microcirculatory disorders are also related to many other diseases, such as diabetes, hypertension, and cerebrovascular diseases. These diseases may cause microcirculatory disorders by inducing microvascular lesions, endothelial dysfunction, inflammatory response and other mechanisms, thereby affecting organ function and disease progression. Therefore, improving microcirculatory disorders is crucial in the treatment of many diseases.

[0003] Currently, there are many drugs to improve microcirculatory disorders, such as antiplatelet drugs, vasodilators or antispasmodics, statins, antioxidants, etc. However, the pathogenesis of microcirculatory disorders is complex, involving multiple pathological and physiological links such as inflammatory response, oxidative stress, vascular endothelial injury, microthrombosis, etc.; the clinical manifestations of microcirculatory disorders are diverse, and the current treatment strategies for microcirculatory disorders have a single target, which is difficult to comprehensively improve microcirculatory disorders and cannot fully meet clinical needs. Finding multi-pathway and multi-target intervention measures is the current research direction.

[0004] In recent years, Chinese medicine has shown certain advantages in the treatment of microcirculatory disorders through various mechanisms, such as blood-activating and blood-stasis-removing drugs such as Danshen, Chuanxiong, Safflower, and Panax notoginseng, warming meridians and dispelling cold drugs such as cinnamon twig and aconite, and Ganoderma lucidum, which can promote blood circulation and reduce platelet aggregation. However, the therapeutic effect of a single Chinese medicine is limited, and multiple Chinese medicines need to be used in combination. The combination method is complicated, and the effect will vary depending on individual differences.

[0005] The combined use of chemical drugs and traditional Chinese medicine can give full play to the rapid effect of chemical drugs and the multi-target and multi-pathway advantages of traditional Chinese medicine, improve microcirculation more comprehensively, and improve the clinical efficacy and quality of life of patients. At present, there is no report showing whether the combined use of vasodilator pancreatic kallikrein and traditional Chinese medicine Ganoderma lucidum can achieve better results in improving microcirculatory disorders. Summary of the invention

[0006] In view of the defects of the prior art, the present invention provides a combined medication of pancreatic kallikreinogenase and Ganoderma lucidum, aiming at better improving vascular microcirculation disorders.

[0007] The present invention provides a combined medicine, which is a vasodilator and ganoderma lucidum administered separately or simultaneously; the weight ratio of the vasodilator to ganoderma lucidum is 0.65-1.3:2000.

[0008] Preferably, the vasodilator is selected from at least one of kallikreinogenase, alprostadil, beraprost sodium, calcium channel blockers, pentoxifylline, sodium nitroprusside, and nitrate vasodilators.

[0009] Preferably, the vasodilator is kallikrein.

[0010] Preferably, the Ganoderma lucidum is selected from Ganoderma lucidum No. 1.

[0011] Preferably, the weight ratio of the vasodilator to Ganoderma lucidum is 0.65:2000 or 1.3:2000.

[0012] Preferably, the weight ratio of the vasodilator to Ganoderma lucidum is 1.3:2000.

[0013] Preferably, the Ganoderma lucidum is a crude drug or an extract thereof, and the extract is a water extract.

[0014] The present invention provides use of any of the above-mentioned combined drugs in the preparation of drugs for treating microcirculatory disorders.

[0015] Preferably, the dosage of the vasodilator is 0.65-1.3 mg / kg; the dosage of the Ganoderma lucidum is 2 g crude drug / kg.

[0016] The present invention provides a pharmaceutical composition, which is prepared by using any of the above-mentioned combined drugs as active ingredients and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0017] The term "vasodilator" refers to a class of chemical or biological drugs that do not include traditional Chinese medicine and its extracts, which can dilate blood vessels and improve blood circulation.

[0018] The present invention screens the ratio of pancreatic kallikreinase and ganoderma lucidum, examines the influence on microcirculatory disorders, and optimizes the best ratio combination for improving microcirculatory function. Experiments show that the combination of pancreatic kallikreinase and ganoderma lucidum can significantly improve the microcirculatory function of mice with microcirculatory disorders, especially when the dosage ratio of pancreatic kallikreinase to ganoderma lucidum is 0.65-1.3:2000, there is a synergistic improvement effect. This provides more options for preventing or treating diseases related to microcirculatory disorders and has good application prospects.

[0019] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0020] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. DETAILED DESCRIPTION

[0021] In the following examples and experimental examples, reagents and materials not particularly described are all commercially available.

[0022] Example 1 Pancreatic kallikreinase combined with Ganoderma lucidum drug This embodiment provides a combined drug of pancreatic kallikreinase and ganoderma lucidum, which can be used simultaneously or separately, and the amount of ganoderma lucidum and pancreatic enzyme used is a mass ratio of 2000:1.3.

[0023] As a preferred scheme, the combined drug contains 2g of Ganoderma Lucidum No. 1 / kg animal and 1.3mg / kg animal of pancreatic kallikreinogenase.

[0024] The preparation method of the combined drug is as follows: 1. Mix The weighed kallikreinogenase, ganoderma lucidum powder and microcrystalline cellulose are placed in a three-dimensional motion mixer and mixed thoroughly.

[0025] 2. Making soft materials Slowly add an appropriate amount of binder solution (hydroxypropyl methylcellulose aqueous solution) to the evenly mixed materials, stirring while adding, so that the materials form a soft material with suitable softness.

[0026] 3. Extrusion The prepared soft material is placed in a screw extruder, a screen with a pore size of 1 mm is selected, and the screw speed and temperature of the extruder are adjusted for extrusion. The extrudate is a cylindrical strip.

[0027] 4. Rounding Immediately transfer the extruded strips to a high-speed centrifugal spheronizer, adjust the speed and spheronization time of the spheronizer, and allow the strips to gradually spheronize to form micropellets under the action of centrifugal force and friction.

[0028] 5. Drying The spheronized pellets are placed in a forced air drying oven and dried at 40-60°C until the moisture content meets the requirements and the moisture content is controlled at 2% - 5%.

[0029] 6. Whole grain The dried micro-pellets may be stuck together or uneven in size. They can be sieved to remove the stuck micro-pellets and fine powder to obtain micro-pellets with a relatively uniform particle size for subsequent enteric coating operations.

[0030] 7. Coating Install and debug the coating pot, turn on the heating and blowing device of the coating pot, and preheat the coating pot to an appropriate temperature. Pour a certain amount of micro-pills carefully into the coating pot, start the coating pot, and let the micro-pills roll slowly in the coating pot to reach a fluidized state. Spray the coating liquid (composed of polyolefin resin II, triethyl citrate, and talcum powder) evenly on the surface of the rolling micro-pills through a spray gun. During the coating process, hot air (temperature 30-40°C) is continuously introduced to dry the micro-pills, so that the solvent in the coating liquid evaporates quickly and promotes the formation of the coating film. Monitor the weight change of the micro-pills in real time, and stop the coating operation when the predetermined weight gain target is reached (coating weight gain 10%-15%).

[0031] Example 2 Pancreatic kallikreinase combined with Ganoderma lucidum drug A combination drug of pancreatic kallikreinase and Ganoderma lucidum is prepared according to the method of Example 1, except that the amount of Ganoderma lucidum and pancreatic enzyme is in a mass ratio of 2000:0.975; as a preferred scheme, the combination drug contains 2g of Ganoderma lucidum No. 1 / kg animal and 0.975mg / kg animal of pancreatic kallikreinase.

[0032] Example 3 Pancreatic kallikreinase combined with Ganoderma lucidum drug A combination drug of pancreatic kallikreinase and Ganoderma lucidum is prepared according to the method of Example 1, except that the amount of Ganoderma lucidum and pancreatic enzyme is in a mass ratio of 2000:0.65; as a preferred scheme, the combination drug contains 2g of Ganoderma lucidum No. 1 / kg animal and 0.65mg / kg animal of pancreatic kallikreinase.

[0033] The technical solution of the present invention is further described below through experiments. The combined drugs used in the following experimental examples are prepared by the methods of the above Examples 1-3.

[0034] Experimental Example 1 Effects of pancreatic kallikreinase combined with Ganoderma lucidum on microcirculatory disorders in mice 1. Experimental Methods 1. Experimental Animals KM mice, 80, half male and half female, weighing 18-22 g, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., Animal Production License No.: SCXK (Beijing) 2019-0010. KM mice were raised in the SPF Animal Laboratory of the Animal Experiment Center of Sichuan Academy of Traditional Chinese Medicine, Animal Use License No.: SYXK (Sichuan) 2023-0100.

[0035] 2. Experimental instruments Microcirculation microscope / microcirculation detection system 3. Animal grouping After 3 days of adaptive feeding, the mice were randomly divided into 7 groups according to their body weight. The groups and drug administration are shown in Table 1.

[0036] Table 1 4. Experimental drugs and their preparation The Ganoderma lucidum used is Medicinal Ganoderma No. 1, which is a variety certified by the Sichuan Non-Major Crop Variety Certification Committee, with the certification number Sichuan Rated Medicinal 2020010, and the fruiting body obtained through cultivation.

[0037] Pancreatic kallikreinase: provided by DeBoer, 70 IU / mg.

[0038] Drugs in each dosing group: (1) A drug of 100% pancreatic kallikreinase + Ganoderma lucidum group was prepared according to the method of Example 1; (2) Prepare the drug of 75% pancreatic kallikreinase + Ganoderma lucidum group according to the method of Example 2; (3) Prepare the drug of 50% pancreatic kallikreinase + Ganoderma lucidum group according to the method of Example 3; (4) According to the preparation method of Example 1, the difference is that the concentration of pancreatic kallikreinase is 0.325 mg / kg, and a drug of 25% pancreatic kallikreinase + Ganoderma lucidum group is prepared; (5) According to the preparation method of Example 1, the difference is that the Ganoderma lucidum powder is not weighed, and a drug of 100% pancreatic kallikreinogenase group is prepared; (6) The preparation method of Example 1 was followed, except that the pancreatic kallikreinogenase was not weighed, to prepare the Ganoderma lucidum group of drugs.

[0039] 5. Animal medication The mice in the normal control group and the model control group were orally administered with distilled water, and the drug-treated group was orally administered according to the group design. The drug was administered once a day for 3 days, with one pill of drug each time. One hour after the last administration, the mice were anesthetized by intraperitoneal injection of chloral hydrate (350 mg / kg), and the mouse hair on the auricle was pulled out with medical adhesive tape. The anesthetized mice were fixed on the mouse observation platform with their abdomen facing down, and the height of the ear support was adjusted so that the mouse's auricle was spread flat on the ear support. 1 to 2 drops of cedar oil were dripped on the auricle, and then the observation platform was placed on the stage of the microscope. The changes in the auricular microcirculation of each group of mice were observed under transmitted light, including the caliber of the auricular arterioles, the caliber of the microvenules, the number of vascular crossings, and the amount of capillary opening.

[0040] Except for the mice in the normal control group, the mice in other groups were intraperitoneally injected with epinephrine (10 μg / 10 g) to create an acute microcirculatory disorder model. The changes in auricular microcirculation of the mice in each group were observed 15 minutes after the injection, including the caliber of auricular arterioles, caliber of microvenules, number of vascular crossings and amount of capillary opening.

[0041] 5. Data processing In this experimental example, there were at least 10 mice in each group. When processing the experimental results of each group, the average value of each index in the group was taken, and the inter-group differences of the indexes in each group were compared by t test.

[0042] The following is the calculation method for the average difference value, average change rate, vascular caliber dilation rate, capillary patency, and increase rate of vascular crossing number: Before and after the injection of adrenaline, the caliber of the auricular arterioles, caliber of the venules, number of vascular crossings and the amount of capillary opening of the mice were measured and recorded.

[0043] Difference value = value before epinephrine injection - value after epinephrine injection; Average difference value = sum of difference values ​​of all mice in the group / number of mice in the group; Change rate = difference value / value before injection of epinephrine × 100%; Average change rate = the sum of the change rates of all mice in the group / the number of mice in the group; Vascular caliber expansion rate = (average difference value of model control group - average difference value of experimental group) ÷ average difference value of model control group × 100%; Increase rate of capillary opening volume = (average difference value of model control group - average difference value of experimental group) ÷ average difference value of model control group × 100%; The increase rate of the number of vascular crossings = (average difference value of the model control group - average difference value of the experimental group) ÷ model average difference value of the control group × 100%.

[0044] 2. Experimental Results By comparing the changes in the microcirculation of the auricle of each group of mice 15 minutes after injection of epinephrine, including the caliber of auricular arterioles and venules, the number of vascular crossings and the amount of capillary opening, we investigated whether the samples have the effect of improving microcirculation. The experimental results are shown in Tables 2-5 below.

[0045] From the experimental results, it can be seen that 15 minutes after the intraperitoneal injection of adrenaline in the model control group mice, the caliber of the venules and arterioles was significantly reduced, the amount of capillary opening was reduced, and the number of vascular crossings was reduced, that is, a significant microcirculatory disorder model was created (p<0.01), proving that the model was successful.

[0046] Table 2 Results of microvenous caliber Table Notes: E represents pancreatic kallikreinogenase, LZ represents Ganoderma lucidum. Compared with the normal group, ** represents P < 0.01; compared with the model group, # represents P < 0.05, ## represents P < 0.01.

[0047] From the results of microvenous caliber in Table 2, it can be seen that the microvenous caliber of mice that took 100% pancreatic kallikrein orally was significantly expanded compared with the model control group mice (p<0.01), and the expansion was 65.1%; the microvenous caliber of mice that took Lingzhi orally did not expand, but contracted, and the expansion ratio was -62.4% compared with the model control group mice; when pancreatic kallikrein and Lingzhi were compounded, the caliber of blood vessels was significantly expanded when the dosage of pancreatic kallikrein was 50%-100%, and the expansion rate reached 65.1%-122.3%, achieving a synergistic improvement effect. Among them, the expansion rate when the dosage of pancreatic kallikrein was 75% was less than the expansion rate of 50%, indicating that when pancreatic kallikrein and Lingzhi were compounded, the effect of the dosage of pancreatic kallikrein on microvenous blood vessels was more complicated, and the more the better, and Lingzhi promoted the role of pancreatic kallikrein in mice.

[0048] Table 3 Results of arteriolar caliber Table Notes: E represents pancreatic kallikreinogenase, LZ represents Ganoderma lucidum. Compared with the normal group, ** represents P < 0.01; compared with the model group, # represents P < 0.05, ## represents P < 0.01.

[0049] From the results of arteriolar caliber in Table 3, it can be seen that the caliber of arteriolar vessels of mice that took 100% pancreatic kallikrein orally was significantly expanded compared with the model control group mice (p<0.01), expanding by 71.9%; the caliber of microvenous vessels of mice that took Lingzhi orally did not expand, but contracted, with an expansion ratio of -19.8% compared with the model control group mice; when pancreatic kallikrein and Lingzhi were compounded, the caliber of blood vessels was significantly expanded when the dosage of pancreatic kallikrein was 50% and 100%, and the expansion rate reached 80.3%-97.1%, achieving a synergistic improvement effect. Among them, the expansion rate when the proportion of pancreatic kallikrein was 75% was less than the expansion rate of 50%, indicating that when pancreatic kallikrein and Lingzhi were compounded, the effect of the dosage of pancreatic kallikrein on arteriolar vessels was more complicated, and the more the better, and Lingzhi promoted the role of pancreatic kallikrein in mice.

[0050] Table 4 Capillary patency results Table Notes: E represents pancreatic kallikreinogenase, LZ represents Ganoderma lucidum. Compared with the normal group, ** represents P < 0.01; compared with the model group, # represents P < 0.05.

[0051] From the results of capillary openness in Table 4, it can be seen that the capillary openness of mice that took 100% kallikrein orally did not change significantly compared with the mice in the model control group, with an increase rate of -1%; the capillary openness of mice that took Ganoderma lucidum orally did not increase, but decreased compared with the mice in the model control group, with an increase rate of -11.1%; when kallikrein was combined with Ganoderma lucidum, the capillary openness increased significantly when the kallikrein dosage was 50%-100%, with an increase rate of 5.74%-59.3%, achieving a synergistic improvement effect.

[0052] Table 5 Results of blood vessel crossing number Table Notes: E represents pancreatic kallikreinogenase, LZ represents Ganoderma lucidum. Compared with the normal group, ** represents P < 0.01; compared with the model group, # represents P < 0.05, ## represents P < 0.01.

[0053] From the results of the vascular crossing number in Table 5, it can be seen that the number of vascular crossings in mice that took 100% kallikrein orally increased compared with the mice in the model control group, with an increase rate of 12.1%; the number of vascular crossings in mice that took Ganoderma lucidum orally increased significantly compared with the mice in the model control group, with an increase rate of 21.2%; when kallikrein and Ganoderma lucidum were combined, the number of vascular crossings increased significantly when the kallikrein dosage was 75%-100%, with an increase rate of 60.6%-76.7%, achieving a synergistic improvement effect.

[0054] In summary, when used in microcirculatory disorder model animals, the combination of pancreatic kallikrein and Ganoderma lucidum No. 1 can improve the expansion of microvenous and arterial calibers, as well as the increase in capillary opening and the number of vascular crossings in mice with microcirculatory disorders. In particular, when 50-100% pancreatic kallikrein was used, the index data was significantly better than that of single oral pancreatic kallikrein or Ganoderma lucidum, and achieved a synergistic improvement effect.

[0055] Through the above embodiments and experimental examples, it can be seen that the present invention screens the ratio of pancreatic kallikreinase and ganoderma lucidum, examines the effect on microcirculatory disorders, and optimizes the best ratio combination for improving microcirculatory function. Experiments have found that the combination of pancreatic kallikreinase and ganoderma lucidum can significantly improve the microcirculatory function of mice with microcirculatory disorders, especially when the dosage ratio of pancreatic kallikreinase to ganoderma lucidum is 0.65-1.3:2000, it has a synergistic improvement effect. This provides more options for preventing or treating diseases related to microcirculatory disorders and has a good application prospect.

Claims

1. A combined drug, characterized in that: The combined drug is a vasodilator and ganoderma lucidum administered separately or simultaneously; the weight ratio of the vasodilator to ganoderma lucidum is 0.65-1.3:2000.

2. The combined drug according to claim 1, characterized in that: The vasodilator is selected from at least one of kallikrein, alprostadil, beraprost sodium, calcium channel blockers, pentoxifylline, sodium nitroprusside, and nitrate vasodilators.

3. The combined drug according to claim 2, characterized in that: The vasodilator is pancreatic kallikrein.

4. The combined drug according to claim 1, characterized in that: The ganoderma lucidum is selected from Ganoderma lucidum No.

1.

5. The combined drug according to claim 1, characterized in that: The weight ratio of the vasodilator to the ganoderma lucidum is 0.65:2000 or 1.3:2000.

6. The combined drug according to claim 5, characterized in that: The weight ratio of the vasodilator to the ganoderma lucidum is 1.3:2000.

7. The combined drug according to claim 1, characterized in that: The ganoderma lucidum is a crude drug or an extract thereof, and the extract is a water extract.

8. Use of the combined drug according to any one of claims 1 to 7 in the preparation of a drug for preventing or treating microcirculatory disorders.

9. Use of the combined drug according to claim 8 in the preparation of a drug for preventing or treating microcirculatory disorders, characterized in that: The dosage of the vasodilator is 0.65-1.3 mg / kg; the dosage of the ganoderma lucidum is 2 g of raw medicine / kg.

10. A pharmaceutical composition, characterized in that: The invention is prepared by taking the combined drug according to any one of claims 1 to 7 as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

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