Application of a composition containing cilostazol to the preparation of a therapeutic agent for cerebrovascular diseases

Abstract

The present invention belongs to the field of pharmacology and relates to the application of a composition of cilostazol and (+)-2-borneol in the preparation of a medicament for treating cerebrovascular disease, particularly ischemic cerebrovascular disease, and more particularly to a composition comprising cilostazol or a pharma- ceutically acceptable salt thereof and (+)-2-borneol, and the application of this composition in the preparation of a medicament for treating cerebrovascular disease, particularly ischemic cerebrovascular disease.

Description

Technical Field

[0001] [Cross-reference] This application claims priority based on Chinese Patent Application No. 202110039436.1, "Application of a Composition Containing Cilostazol to Cerebrovascular Diseases", filed with the China National Intellectual Property Administration on January 13, 2021, the entire content of which is incorporated herein by reference.

[0002] The present invention belongs to the field of pharmacy and relates to the application of a composition of cilostazol and (+)-2-borneol in the preparation of a medicament for treating cerebrovascular diseases, particularly ischemic cerebrovascular diseases.

Background Art

[0003] Cerebrovascular disease (CVD) refers to brain lesions caused by various cerebrovascular diseases, and is divided into acute cerebrovascular diseases (stroke) and chronic cerebrovascular diseases according to its onset process. Acute cerebrovascular diseases include transient ischemic attack, cerebral thrombosis, cerebral embolism, hypertensive encephalopathy, cerebral hemorrhage, and subarachnoid hemorrhage. Chronic cerebrovascular diseases include cerebral arteriosclerosis, cerebrovascular dementia, cerebral artery steal syndrome, Parkinson's disease, etc. Ischemic stroke refers to the general term for necrosis of brain tissue caused by stenosis or occlusion of the arteries supplying blood to the brain (carotid artery and vertebral artery) and insufficient blood supply to the brain. Cerebral ischemia includes four types: transient ischemic attack (TIA), reversible ischemic neurological deficit (RIND), progressive stroke (SIE), and complete stroke (CS). No cerebral infarction is observed in TIA, but different degrees of cerebral infarction are observed in RIND, SIE, and CS.

[0004] Cilostazol is a drug for suppressing platelet aggregation. It was first developed and synthesized by Otsuka Pharmaceutical Co., Ltd. in Japan. It was also launched in Japan in 1988, obtained the approval of the US FDA in May 1999, and was imported into China in 1996. Cilostazol is a selective inhibitor of type 3 phosphodiesterase (PDE3). The binding rate of cilostazol to plasma proteins is approximately 95%, and most of them exist as relatively stable prototypes. Cilostazol has a wide range of pharmacological activities and has clinical value for many diseases, such as peripheral thrombotic diseases and intermittent claudication. And cilostazol can prevent circulatory shock and coronary artery restenosis by having antiplatelet and vasodilatory effects. According to research, PDE3 has been shown to inhibit the degradation of cAMP in the circulatory system, increase cAMP in platelets and vascular smooth muscle, inhibit platelet formation, and also promote the proliferation of vascular smooth muscle cells. Cilostazol mainly inhibits platelet degradation by affecting factors such as arachidonic acid, adenosine diphosphate, epinephrine, collagen, and fibrinolase. Currently, experts recommend treating patients with carotid artery thrombosis with cilostazol, which can thereby enable the treatment or prevention of cerebral ischemia. In addition, PDE3 can inhibit the production of nitric oxide synthase (NOS), thereby reducing the production of nitric oxide (NO).

[0005] The structural formula of cilostazol is as follows.

[0006] [Chemical formula]

[0007] The main component of natural borneol is stipulated in the "Chinese Pharmacopoeia" 2020 edition that the content of (+)-2-borneol in natural borneol must be 96% or more. (+)-2-borneol is a bicyclic monoterpene compound present in the volatile oils of many traditional Chinese medicines, and exhibits various physiological activities such as anti-inflammatory, antioxidant, and enhancement of GABA receptor function (Euro J Pharma 2017, 811: 1-11). Borneol has been approved by the US FDA as a food flavor or adjuvant (21 CFR 172.515). Also, borneol is an oral adjuvant used to treat many diseases, and the recommended oral dose of natural borneol for adults is 0.3 - 0.9 g / day in the "Chinese Pharmacopoeia" 2020 edition. In addition, Xingnaojing injection (which contains about 1 mg / mL of borneol and is in Volume 17 of the Ministry of Health's Chinese Medicine Standards Prescription) is clinically diluted with 250 - 500 mL of 5% - 10% glucose injection or sodium chloride injection every 10 - 20 mL and administered by intravenous drip, so it is estimated that the dose of borneol required for a single intravenous drip injection is 10 - 20 mg. Also, borneol is used as an important component of Bingpeng San, and its content must be 30 mg / g (3%) or more in the "Chinese Pharmacopoeia" 2020 edition.

[0008] Therefore, providing the application of a composition containing cilostazol to cerebrovascular diseases has important practical significance.

Summary of the Invention

Problems to be Solved by the Invention

[0009] An object of the present invention is to provide the application of a pharmaceutical composition containing cilostazol or a pharmaceutically acceptable salt thereof and (+)-2-borneol in the preparation of a therapeutic agent for cerebrovascular diseases. By combining these drugs, the therapeutic effect on cerebrovascular diseases can be synergistically enhanced.

Means for Solving the Problems

[0010] The composition provided by the present invention comprises the following components: Component (I) which is cilostazol, its derivative, its pharmaceutically acceptable salt, or its stereoisomer, or its prodrug molecule, and contains component (II) which is (+)-2-borneol, borneol, or a drug having (+)-2-borneol as an active ingredient.

[0011] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 100:1 to 1:1.

[0012] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 50:1 to 1:1.

[0013] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 36:1 to 1:1.

[0014] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 36:1 to 3:1.

[0015] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 18:1 to 3:1.

[0016] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 9:1 to 3:1.

[0017] In some specific embodiments of the present invention, the weight ratio of the component (I) to the component (II) is 1:1, 3:1, 9:1, 10:1 and / or 18:1.

[0018] Furthermore, the present invention provides a drug comprising the composition and a pharmaceutically acceptable additive.

[0019] Based on the above research, the present invention also provides the use of the composition or the drug in the preparation of a drug for preventing and / or treating cerebrovascular diseases.

[0020] In some specific embodiments of the present invention, the cerebrovascular diseases include ischemic cerebrovascular diseases, and preferably, the ischemic cerebrovascular diseases include ischemic stroke.

[0021] In some specific embodiments of the present invention, according to the composition, it can significantly improve the neuropathy in MCAO mice and reduce the cerebral infarction size.

[0022] In the present invention, cerebrovascular disease (CVD) refers to brain lesions caused by various cerebrovascular diseases, and may be classified into two types: acute cerebrovascular disease (stroke) and chronic cerebrovascular disease according to its onset process. Examples of acute cerebrovascular diseases include transient ischemic attack, cerebral thrombosis, cerebral embolism, hypertensive encephalopathy, cerebral hemorrhage, and subarachnoid hemorrhage. Examples of chronic cerebrovascular diseases include cerebral arteriosclerosis, cerebrovascular dementia, cerebral artery steal syndrome, Parkinson's disease, etc. Ischemic stroke refers to the general term for necrosis of brain tissue caused by stenosis or occlusion of the arteries supplying blood to the brain (carotid artery and vertebral artery) and insufficient blood supply to the brain. Cerebral ischemia includes four types: transient ischemic attack (TIA), reversible ischemic neurological deficit (RIND), progressive stroke (SIE), and complete stroke (CS). No cerebral infarction is observed in TIA, but different degrees of cerebral infarction are observed in RIND, SIE, and CS.

[0023] By administering cilostazol at 1 to 18 mg / kg or (+)-2-borneol at 0.27 to 5 mg / kg via the tail vein to a rat model of local cerebral ischemia-reperfusion injury, the present inventors found that the neurological deficits in MCAO rats were significantly improved, the area of cerebral infarction could be reduced, and when components (with the mass ratio of cilostazol:(+)-2-borneol being 18:1 to 1:1) were combined within the above dosage range, a synergistic effect could be achieved. By administering cilostazol at 10 to 19.46 mg / kg or (+)-2-borneol at 0.54 to 10 mg / kg via the vein to a mouse model of local cerebral ischemia-reperfusion injury, the neurological deficits can be significantly improved and the area of cerebral infarction can be reduced.

Mode for Carrying Out the Invention

[0024] The present invention discloses the application of a composition containing cilostazol to cerebrovascular diseases. However, those skilled in the art can refer to the content of this specification and appropriately improve the process parameters to implement the present invention. It should be noted that all similar substitutions and modifications are obvious to those skilled in the art, and they should be regarded as being included in the present invention. Although the methods and applications according to the present invention have been described using preferred examples, it is obvious that those skilled in the art can modify or make appropriate changes and combinations to the methods and applications described in this specification without departing from the content, idea, and scope of the present invention in order to implement and apply the technology of the present invention.

[0025] In the pharmaceutical composition provided by the present invention, the weight ratio of cilostazol or its pharmaceutically acceptable salt:(+)-2-borneol is 100:1 to 1:1, preferably the weight ratio is 50:1 to 1:1, preferably the weight ratio is 36:1 to 1:1, preferably the weight ratio is 36:1 to 3:1, preferably the weight ratio is 18:1 to 3:1 and 9:1 to 3:1. More preferably, the weight ratio is 1:1, 3:1, 9:1, 10:1 and / or 18:1.

[0026] The drug combination of the present invention can be applied to the preparation of drugs for cerebrovascular diseases. Here, the cerebrovascular disease is preferably an ischemic cerebrovascular disease, and more preferably an ischemic stroke.

[0027] According to the present invention, by combining cilostazol and (+)-2-borneol, an advantageous effect was obtained in the pharmacological efficacy test of animals (rats, mice) that the pharmacological efficacy against cerebrovascular diseases could be synergistically enhanced.

[0028] Any of the raw materials and reagents used in the application of the composition containing cilostazol provided by the present invention to cerebrovascular diseases can be purchased from the market. Hereinafter, the present invention will be further described with reference to examples.

Example

[0029] Example 1 Research 1 on the protective effect of the composition of cilostazol and (+)-2-borneol on focal cerebral ischemia-reperfusion injury

[0030] 1. Materials and methods 1.1 Experimental animals Sprague-Dawley (SD) rats, male, SPF grade, body weight 250-280 g. 1.2 Test drugs

[0031]

Table 1

[0032] 1.3 Experimental methods 1.3.1 Grouping and dosing of animals The experimental animals were divided into four groups: cilostazol group (1 mg / kg), (+)-borneol group (1 mg / kg), composition group of cilostazol and (+)-borneol (2 mg / kg, cilostazol:(+)-borneol = 1:1, cilostazol 1 mg / kg, (+)-borneol 1 mg / kg), and model group. After creating a cerebral ischemia model, the animals were assigned to each group with equal probability by single-blind method. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal amount of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, photographed, and the area of cerebral infarction was measured.

[0033] 1.3.2 Establishment of local cerebral ischemia-reperfusion model A rat local cerebral ischemia-reperfusion model was established by the internal carotid artery suture method. The limbs and head of anesthetized rats were tied with rubber bands (the hind limbs were fixed above the knee joint and the forelimbs were fixed below the wrist joint), the animals were fixed in the supine position on the operating table, the hair from the head to the chest was shaved with an animal shaver, and the skin was disinfected with alcohol. The neck was incised along the midline, and the subcutaneous tissue was bluntly dissected. The thin fascia on the surface of the anterior triangle of the neck was dissected, the lower end of the omohyoid muscle was pulled out, and an artery pulsating vertically parallel to this muscle was visible. The sheath of the artery was opened to expose the right carotid artery bifurcation, the right common carotid artery, external carotid artery, and internal carotid artery were dissected, the vagus nerve was gently dissected, and the external carotid artery was ligated and cut. The proximal end of the common carotid artery was occluded, an incision was made from the distal end of the external carotid artery ligation, an embolization wire was inserted, passed through the bifurcation of the common carotid artery into the internal carotid artery, and then slowly inserted until resistance was felt (about 20 mm from the bifurcation), completely blocking the blood supply to the middle cerebral artery. The embolization wire was slightly fixed under the incision of the external carotid artery with a thread, the thread occluding the proximal end of the common carotid artery was loosened, the wound was covered with a gauze soaked in sterile physiological saline, and the rat was placed on a heating pad for warming. After 2.0 hours of cerebral ischemia on the right side, the embolization wire was slowly withdrawn to restore blood supply and reperfusion was performed. The external carotid artery was ligated with the thread of the fixed embolization wire, the skin was sutured and disinfected. The rats were placed in clean feed, and their normal state and breathing were observed until they woke up from anesthesia. Feed and water were given, and they were bred by the normal method.

[0034] 1.3.3 Measurement of the cerebral infarction area After evaluating the neurological deficit symptoms of the animals, they were sacrificed with CO2. After decapitation, the brains were removed, and the olfactory bulbs, cerebellum, and lower brainstem were removed. The blood on the brain surface was washed with physiological saline, the remaining water on the surface was absorbed, and the brains were placed at -20°C for 20 min. Immediately after removal, coronal sections were cut at 2-mm intervals with an intersecting plane perpendicular to the line of sight, and then stained in a 1% TTC solution (37°C, 30 min). Normal brain tissue was stained dark red, and ischemic brain tissue was lightly stained. After washing with physiological saline, the brain sections were quickly arranged in a row from the front to the back, the remaining water on the surface was absorbed, and the surface was dried. Next, photographs were taken.

[0035] Calculation of the cerebral infarction area: The photographs were processed with Image J software, and the corresponding areas of the left brain and the non-infarcted areas of the right brain were determined according to the following formula, and the ratio of the infarction range was calculated.

[0036] Calculation method of the infarction volume: V=t(A1+A2+A3+……+An) where t is the section thickness and A is the infarction area.

[0037] %I=100%×(VC-VL) / VC where %I is the ratio of the infarction volume, VC is the brain volume of the target side (left hemisphere), and VL is the volume of the non-infarcted area of the infarcted side (right hemisphere).

[0038] 1.3.4 Analysis of the synergism of the composition According to Kim's formula q = E(a + b) / (Ea + Eb - Ea×Eb), it was evaluated whether cilostazol and (+)-borneol in the composition had a synergistic effect. In the formula, E(a + b) is the effective rate of the combined drug, and Ea and Eb are the effective rates when drug A (cilostazol) and drug B ((+)-borneol) are administered alone, respectively. E administration group=(X モデル -X 投与 ) / X モデルHowever, X is the value of the cerebral infarction range. When the q value is within the range of 0.85 to 1.15, it indicates that the combination of the two drugs has a simple additive effect. When the q value > 1.15, it indicates that the combination of the two drugs has a synergistic effect. When the q value < 0.85, it indicates that the combination of the two drugs has an antagonistic effect.

[0039] 1.4 Statistics of Data The experimental data were expressed as mean ± standard deviation (Mean ± SD). The differences between groups were analyzed by one-way analysis of variance, and the comparison between groups was tested by the LSD method. P < 0.05 was defined as having a significant difference.

[0040] 2. Experimental Results The influence on the range of cerebral infarction is shown in Table 1. As a result of the experiment, the administration of (+)-2-borneol 1 mg / kg and the composition (cilostazol 1 mg / kg + (+)-2-borneol 1 mg / kg) could significantly reduce the range of cerebral infarction in animals (p < 0.01, p < 0.001). The administration of cilostazol 1 mg / kg tended to improve cerebral ischemia injury, but there was no statistical difference (p = 0.06). The calculation result of synergy was q = 1.24, indicating that the combination of the two drugs produced a synergistic effect.

[0041]

Table 2

[0042] Study 2 on the Protective Effect of the Composition of Cilostazol and (+)-2-Borneol against Focal Cerebral Ischemia-Reperfusion Injury

[0043] 1. Materials and Methods 1.1 Experimental Animals Sprague-Dawley (SD) rats, male, SPF grade, body weight 250 - 280 g.

[0044] 1.2 Test Drugs Cilostazol and (+)-2-borneol were the same as in Example 1.

[0045] 1.3 Experimental methods The experimental animals were divided into four groups: cilostazol group (9 mg / kg), (+)-2-borneol group (1 mg / kg), composition group of cilostazol and (+)-2-borneol (10 mg / kg, cilostazol:(+)-2-borneol = 9:1, cilostazol 9 mg / kg + (+)-2-borneol 1 mg / kg), and model group. After establishing the cerebral ischemia model, the animals were assigned to each group with equal probability by single-blind method. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal volume of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, and photographed to measure the range of cerebral infarction.

[0046] The establishment of the local cerebral ischemia-reperfusion model, the measurement of the range of cerebral infarction, the analysis of the synergy of the composition, and the statistical methods of the data were the same as those in Example 1.

[0047] 2. Experimental results Table 2 shows the effects on the range of cerebral infarction. From the experimental results, the administration of cilostazol 9 mg / kg, (+)-2-borneol 1 mg / kg, and the composition (cilostazol 9 mg / kg + (+)-2-borneol 1 mg / kg) could significantly reduce the range of cerebral embolism in animals (p = 0.024, p = 0.017, p = 0.000). The calculation result of synergy was q = 1.5, indicating that the combined use of the two drugs produced a synergistic effect.

[0048]

Table 3

[0049] Example 3 Research on the protective effect of the composition of cilostazol and (+)-2-borneol on local cerebral ischemia-reperfusion injury 3

[0050] 1. Materials and methods 1.1 Experimental animals Sprague-Dawley (SD) rats, male, SPF grade, body weight 250 - 280 g.

[0051] 1.2 Test agent Cilostazol and (+)-2-borneol were the same as in Example 1.

[0052] 1.3 Experimental method The experimental animals were divided into four groups: the cilostazol group (18 mg / kg), the (+)-2-borneol group (1 mg / kg), the composition group of cilostazol and (+)-2-borneol (19 mg / kg, cilostazol:(+)-2-borneol = 18:1, cilostazol 18 mg / kg + (+)-2-borneol 1 mg / kg), and the model group. After creating a cerebral ischemia model, the animals were assigned to each group with equal probability by a single-blind method. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal volume of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, and photographed to measure the area of cerebral infarction.

[0053] The creation of the local cerebral ischemia-reperfusion model, the measurement of the area of cerebral infarction, the analysis of the synergy of the composition, and the statistical method of the data were the same as in Example 1.

[0054] 2. Experimental results Table 3 shows the effects on the area of cerebral infarction. From the experimental results, it was shown that the administration of cilostazol 18 mg / kg, (+)-2-borneol 1 mg / kg, and the composition (cilostazol 18 mg / kg + (+)-2-borneol 1 mg / kg) could significantly reduce the area of cerebral embolism in animals (p = 0.001, p = 0.002, p = 0.000). The calculation result of synergy was q = 1.24, indicating that the combined use of the two drugs produced a synergistic effect.

[0055]

Table 4

[0056] Example 4 Effects of cilostazol / (+)-2-borneol (1:1, 3:1, 9:1) on local cerebral ischemia-reperfusion injury

[0057] 1. Materials and Methods 1.1 Experimental Animals Sprague-Dawley (SD) rats, male, SPF grade, weighing 250 - 280 g.

[0058] 1.2 Test Drugs Cilostazol and (+)-borneol were the same as in Example 1.

[0059] 1.3 Experimental Methods The experimental animals were divided into four groups: a model group and three groups of cilostazol / (+)-borneol compositions (1:1 group, cilostazol 5 mg / kg + (+)-borneol 5 mg / kg; 3:1 group, cilostazol 7.5 mg / kg + (+)-borneol 2.5 mg / kg; 9:1 group, cilostazol 9 mg / kg + (+)-borneol 1 mg / kg. The total administered dose of each composition was 10 mg / kg). After establishing a cerebral ischemia model, the animals were randomly assigned to each group with equal probability by single-blind method. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal volume of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, and photographed to measure the area of cerebral infarction.

[0060]

[0061] 2. Experimental Results The effect on the area of cerebral infarction is shown in Table 4. From the experimental results, it was shown that the combined administration of cilostazol / (+)-borneol at 1:1, 3:1, and 9:1 could significantly reduce the area of cerebral embolism in animals (p < 0.001).

[0062] [Table 5]

[0063] ​Example 5 Effects of Cilostazol / (+)-2-Borneol (9:1, 18:1, 36:1) on Focal Cerebral Ischemia-Reperfusion Injury

[0064] 1. Materials and Methods 1.1 Experimental Animals Sprague-Dawley (SD) rats, male, SPF grade, body weight 250 - 280 g.

[0065] 1.2 Test Drugs Cilostazol and (+)-2-borneol were the same as in Example 1.

[0066] 1.3 Experimental Methods The experimental animals were divided into four groups: a model group and three cilostazol / (+)-2-borneol composition groups (9:1 group, cilostazol 9 mg / kg + (+)-2-borneol 1 mg / kg; 18:1 group, cilostazol 9.47 mg / kg + (+)-2-borneol 0.53 mg / kg; 36:1 group, cilostazol 9.73 mg / kg + (+)-2-borneol 0.27 mg / kg. The total administered dose of each composition was 10 mg / kg). After establishing the cerebral ischemia model, the animals were randomly assigned to each group by single-blind method with equal probability. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal volume of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, and photographed to measure the area of cerebral infarction.

[0067] The establishment of the focal cerebral ischemia-reperfusion model, the measurement of the area of cerebral infarction, and the statistical method of data were the same as in Example 1.

[0068] 2. Experimental Results Table 5 shows the effects on the area of cerebral infarction. From the experimental results, it was shown that the combined administration of cilostazol / (+)-2-borneol at 9:1, 18:1, and 36:1 could significantly reduce the area of cerebral embolism in animals (p < 0.001).

[0069]

Table 6

[0070] Example 6 Effect of Cilostazol / (+)-2-Borneol (1:1, 3:1, 9:1) on Focal Cerebral Ischemia-Reperfusion Injury in Mice

[0071] 1. Materials and Methods 1.1 Experimental Animals C57BL / 6J mice, male, SPF grade, 8 weeks old.

[0072] 1.2 Test Drugs Cilostazol and (+)-2-borneol were the same as in Example 1.

[0073] 1.3 Experimental Methods The experimental animals were divided into four groups: a model group and three groups of cilostazol / (+)-2-borneol compositions (1:1 group, cilostazol 10 mg / kg + (+)-2-borneol 10 mg / kg; 3:1 group, cilostazol 15 mg / kg + (+)-2-borneol 5 mg / kg; 9:1 group, cilostazol 18 mg / kg + (+)-2-borneol 2 mg / kg. The total administered dose of each composition was 20 mg / kg). After establishing the cerebral ischemia model, the animals were assigned to each group with equal probability by single-blind method. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal volume of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, and photographed to measure the area of cerebral infarction.

[0074] Establishment of a local cerebral ischemia-reperfusion model: A local cerebral ischemia-reperfusion model in mice was established by the internal carotid artery suture method. The anesthetized mice were fixed on the operating table in the supine position. The neck was incised along the midline, and the subcutaneous tissue was bluntly dissected. The right common carotid artery, external carotid artery, and internal carotid artery were dissected. An embolization wire was inserted from the external carotid artery, passed through the bifurcation of the common carotid artery, and entered the internal carotid artery. Then, it was slowly inserted until resistance was felt (about 10 mm from the bifurcation), and the blood supply to the middle cerebral artery was completely blocked. After 60 minutes of right cerebral ischemia, the embolization wire was slowly withdrawn to restore blood supply and reperfusion was performed. The mice were placed in clean feed, and their respiration was observed in the normal state until they woke up from anesthesia. Feed and water were added, and they were bred by the normal method.

[0075] The measurement method of the range of cerebral embolism and the statistical method of data were the same as those in Example 1.

[0076] 2. Experimental results Table 6 shows the effect on the range of cerebral infarction. From the experimental results, it was shown that the combined administration of cilostazol / (+)-borneol at 1:1, 3:1, and 9:1 could significantly reduce the range of cerebral embolism in animals (p<0.001).

[0077]

Table 7

[0078] Example 7 Effects of cilostazol / (+)-borneol (9:1, 18:1, 36:1) on local cerebral ischemia-reperfusion injury in mice

[0079] 1. Materials and methods 1.1 Experimental animals C57BL / 6J mice, male, SPF grade, 8 weeks old.

[0080] 1.3 Experimental methods The experimental animals were divided into four groups: a model group and three groups of cilostazol / (+)-2-borneol compositions (9:1 group, cilostazol 18 mg / kg + (+)-2-borneol 2 mg / kg; 18:1 group, cilostazol 18.95 mg / kg + (+)-2-borneol 1.05 mg / kg; 36:1 group, cilostazol 19.46 mg / kg + (+)-2-borneol 0.54 mg / kg. The total dosage of each composition administered was 20 mg / kg). After establishing a cerebral ischemia model, the animals were randomly assigned to each group with equal probability by a single-blind method. The animals were administered the drug intravenously once immediately after reperfusion, while the animals in the model group were administered an equal volume of physiological saline. At 24 hours after cerebral ischemia, the animals were sacrificed, the brains were removed, stained, and photographed to measure the area of cerebral infarction.

[0081] The establishment of the local cerebral ischemia-reperfusion model was the same as in Example 6, and the measurement of the area of cerebral embolism and the statistical method of the data were the same as in Example 1.

[0082] 2. Experimental Results Table 7 shows the effect on the area of cerebral infarction. From the experimental results, it was shown that the combined administration of cilostazol / (+)-2-borneol at ratios of 9:1, 18:1, and 36:1 could significantly reduce the area of cerebral embolism in animals (p < 0.001, p < 0.001, p < 0.01).

[0083]

Table 8

[0084] As described above in detail, the application of the composition containing cilostazol provided by the present invention to cerebrovascular diseases has been described. The principle and embodiments of the present invention are described herein using specific examples, and the descriptions in the above examples are only used to understand the method and gist of the present invention. It should be noted that those skilled in the art may make several improvements and modifications to the present invention without departing from the principle of the present invention, and these improvements and modifications also fall within the scope of the patent of the present invention.

Claims

1. A pharmaceutical composition for preventing and / or treating cerebrovascular diseases, comprising component (I) which is cilostazol, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and component (II) which is (+)-2-borneol, and not containing active ingredients other than component (I) and component (II), wherein the weight ratio of component (I) to component (II) is 100:1 to 1:

1.

2. The pharmaceutical composition according to claim 1, wherein the weight ratio of component (I) to component (II) is 50:1 to 1:

1.

3. The pharmaceutical composition according to claim 1, wherein the weight ratio of component (I) to component (II) is 36:1 to 1:

1.

4. The pharmaceutical composition according to claim 1, wherein the weight ratio of component (I) to component (II) is 36:1 to 3:

1.

5. The pharmaceutical composition according to claim 1, wherein the weight ratio of component (I) to component (II) is 18:1 to 3:

1.

6. The pharmaceutical composition according to claim 1, wherein the weight ratio of component (I) to component (II) is 9:1 to 3:

1.

7. The pharmaceutical composition according to claim 1, wherein the weight ratio of component (I) to component (II) is 1:1, 3:1, 9:1, 10:1, or 18:

1.

8. A medicament, comprising the pharmaceutical composition according to any one of claims 1 to 7 and a pharmaceutically acceptable additive.

9. Use of the pharmaceutical composition according to any one of claims 1 to 7 or the medicament according to claim 8 in the manufacture of a medicament for preventing and / or treating cerebrovascular diseases.

10. The use according to claim 9, wherein the cerebrovascular disease is an ischemic cerebrovascular disease.

11. The use according to claim 10, wherein the ischemic cerebrovascular disease is ischemic stroke.

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