Application of novel glycyrrhizol in preparation of product for preventing or treating ischemic encephalopathy

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of novel glycyrrhizol in preparation of products for preventing or treating ischemic encephalopathy, and the ischemic encephalopathy comprises cerebral thrombosis, cerebrovascular stenosis / occlusion and cerebral atherosclerosis. According to the application disclosed by the invention, the novel glycyrrhizol is used for treating the ischemic encephalopathy for the first time, and experiments show that the novel glycyrrhizol has a remarkable treatment effect on the ischemic encephalopathy, can be used for obviously improving behavioral scores, cerebral thrombosis and vascular diameter conditions after ischemia and reducing the volume of cerebral infarction, and has a good treatment effect on cerebral thrombosis and middle artery embolism. Therefore, the compound can be used for preparing medicines for preventing or treating ischemic encephalopathy.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to a new use of new glycyrrhetol in preparation of a product for preventing or treating ischemic encephalopathy. BACKGROUND

[0002] Stroke, also known as apoplexy, is an acute cerebrovascular disease, and is one of the key diseases leading to most adult deaths and disabilities. Stroke is generally divided into two categories, one of which is ischemic stroke, which has the highest incidence, accounting for about 87% of all stroke cases; the other is hemorrhagic stroke, which accounts for a smaller part but has a higher mortality rate and more serious clinical outcomes. The pathogenesis of ischemic stroke is mainly due to blood stasis and the resulting cut-off of energy supply in the perfusion area, thereby causing damage to the nervous system, such as cerebral infarction, cerebral edema and inflammatory reaction. Intravenous injection of recombinant tissue plasminogen activator (tPA) or interventional thrombectomy / stenting within the time window is the most effective treatment method for patients with ischemic stroke in the acute phase, which can save part of the neuronal function. However, more than 90% of ischemic stroke patients cannot receive reperfusion therapy due to the strict time window (thrombolysis 3.5 to 4.5 hours, thrombectomy 6 hours). Even for patients who receive reperfusion therapy, more than 40% of patients still have severe complications and long-term disability, such as language disorders, hemiplegia, cognitive impairment and dependence on daily activities. In addition, due to the limited access to medical resources, high cost and limited regenerative capacity of the central nervous system, neurorehabilitation, which is considered as the main treatment method for stroke, can only benefit a small number of stroke patients. On the other hand, hemorrhagic stroke is the result of non-traumatic vascular rupture, leading to intracranial hematoma. Surgical removal of hematoma is considered to be the key to improving the mortality rate of hemorrhagic stroke. However, for both ischemic stroke and hemorrhagic stroke, there is no effective treatment method once the neural function damage occurs. Since the pathogenesis of stroke is not fully understood, there are almost no clinical treatment strategies except tPA. Therefore, it is of great significance to seek effective prevention and treatment methods for stroke. SUMMARY

[0003] The purpose of the present application is to overcome the shortcomings of the prior art and provide a new use of new glycyrrhetol in treating ischemic encephalopathy.

[0004] In order to achieve the above-mentioned purpose, the present application provides the use of new glycyrrhetol in the preparation of a product for preventing or treating ischemic encephalopathy.

[0005] The chemical structural formula of the new glycyrrhetol is as follows:

[0006] .

[0007] The ischemic encephalopathy according to the present application includes cerebral thrombosis, cerebral vascular stenosis / occlusion, cerebral atherosclerosis, etc.

[0008] The product includes a drug or a food.

[0009] The food includes a functional food or a health product.

[0010] The dosage form of the drug includes, but is not limited to, an injection, a capsule, a tablet, a granule, a gel, a sustained-release preparation, an oral liquid, a dripping pill or a nano preparation. The drug includes a pharmaceutically acceptable excipient, and the excipient includes a filler, a disintegrant, a lubricant, a suspending agent, a binder, a sweetener, a flavoring agent, a preservative, a base, etc. The filler includes starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.; the disintegrant includes starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linked sodium carboxymethyl cellulose, etc.; the lubricant includes magnesium stearate, sodium dodecyl sulfate, talc, silicon dioxide, etc.; the suspending agent includes polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methyl cellulose, etc.; and the binder includes starch paste, polyvinylpyrrolidone, hydroxypropyl methyl cellulose, etc.

[0011] The new glycyrrhetol is a commercially available product.

[0012] Compared with the prior art, the present application first uses the new glycyrrhetol to treat ischemic encephalopathy. Experiments show that the new glycyrrhetol has a significant therapeutic effect on ischemic encephalopathy, can obviously improve the behavior score after ischemia, cerebral thrombosis and blood vessel diameter, reduce the cerebral infarction volume, and has a good therapeutic effect on cerebral thrombosis and middle artery embolism. Therefore, the new glycyrrhetol can be used to prepare a drug for preventing or treating ischemic encephalopathy. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 The figure is a schematic diagram of the experimental results of the influence of the new glycyrrhetol according to the present application on the brain thrombosis of zebrafish.

[0014] Figure 2 The figure is a schematic diagram of the experimental results of the influence of the new glycyrrhetol according to the present application on the movement of zebrafish with brain thrombosis, wherein A is a zebrafish movement trajectory diagram; B is the total distance of movement, C is the average speed, D is the activity, E is the acceleration, F is the angular velocity, and G is the movement sinuosity. * P<0.05, ** P<0.01, *** P<0.001.

[0015] Figure 3The experimental results of the influence of the new glycyrrhetinol involved in the application on the blood vessel diameter of zebrafish are shown in the schematic diagram, wherein A is a blood vessel fluorescence picture; (B) is a blood vessel diameter statistical analysis diagram; *P<0.05, **P<0.01, ***P<0.001.

[0016] Figure 4 The result schematic diagram of the new glycyrrhetinol involved in the application for improving the cerebral infarction of mice. DETAILED DESCRIPTION

[0017] The application will be further described in detail below by specific examples and in combination with the drawings.

[0018] Example 1

[0019] This example relates to the experiment of the effect of the new glycyrrhetinol on cerebral thrombosis, and the new glycyrrhetinol is purchased from the United States MedChemExpress company, and the specific experimental steps are as follows:

[0020] First, the influence of the new glycyrrhetinol on the thrombus area and motor impairment of zebrafish with cerebral thrombosis

[0021] 1. Establishing a zebrafish cerebral thrombosis model

[0022] Zebrafish is a good model organism for studying thrombosis. Ponatinib is used to prepare a zebrafish cerebral ischemia model, specifically: 2 dpf zebrafish embryos are placed in a 6-well plate for culture, 15-20 embryos per well. After the zebrafish larvae are developed, the zebrafish larvae are treated with 1 μg / ml ponatinib (pon) for 24 hours to induce cerebral thrombosis, and the cerebral thrombosis model zebrafish is obtained.

[0023] 2. The influence of the new glycyrrhetinol on zebrafish cerebral thrombosis

[0024] After the model zebrafish is incubated with the new glycyrrhetinol (1 μg / ml) for 24 hours, it becomes the experimental group; the normal zebrafish larvae are used as the normal control group; the model zebrafish treated with aspirin (1 μg / ml) is used as the positive control group; the zebrafish is dyed using o-dianisidine dyeing agent, and each group of zebrafish is placed in 0.6 mg / ml o-dianisidine dyeing liquid containing 10 mM sodium acetate and 4v / v% ethanol, incubated in the dark at 28 ℃ for 15 min, and then washed with 100% DMSO (dimethyl sulfoxide) for 3 times. The zebrafish is observed and photographed under a stereomicroscope. The results are shown in Figure 1

[0025] From Figure 1 ​It can be seen that stimulation of juvenile zebrafish with 1 μg / ml ponatinib caused significant cerebral thrombosis and a significant reduction in cardiac blood perfusion, suggesting that ponatinib can obstruct systemic circulation, indicating successful modeling. Compared with the model group, zebrafish treated with neoglycyrrhizin showed a significant reduction in thrombosis and a significant increase in myocardial hemoglobin staining intensity, indicating that neoglycyrrhizin has a significant therapeutic effect on zebrafish thrombosis. The therapeutic effect of neoglycyrrhizin on zebrafish with cerebral thrombosis is comparable to that of aspirin.

[0026] 3. Zebrafish sports injury assessment

[0027] Zebrafish from the normal control group, the model group, and the experimental group treated with different concentrations (4 μg / ml, 2 μg / ml, 1 μg / ml, 0.5 μg / ml) of neoglycyrrhizin were placed in 96-well plates, one fish per well. The zebrafish were acclimatized in the 96-well plates for 10 min at 28℃ before movement was recorded. All experiments were conducted in bright or dark environments (10 min bright, 10 min dark alternation) for a total of 60 min. The total distance, average velocity, acceleration, angular velocity, and activity level of the zebrafish were recorded and analyzed using a viewpoint behavior analyzer. The zebrafish behavior trajectory was tracked and quantified using the Nordas zebrafish behavior trajectory tracking system. The results are as follows: Figure 2 As shown.

[0028] from Figure 2 As can be seen, compared with the normal control group, the zebrafish in the model group exhibited significantly reduced total distance of movement, average speed, acceleration, and activity frequency, as well as significantly increased flexure and angular velocity, indicating that cerebral thrombosis can cause damage to the overall behavior and neurobehavioral behavior of zebrafish. Compared with the model group, these motor indicators were significantly reversed in all four experimental groups, indicating that neoglycyrrhizin can improve the motor damage in zebrafish caused by cerebral ischemia.

[0029] II. Effects of Neoglycyrrhizin on the Vascular System of Zebrafish with Cerebral Thrombosis

[0030] Using genetically modified zebrafish Tg ( fli1 Using EGFP as the experimental subject, juvenile zebrafish were stimulated with ponatinib for 24 hours as the model group; the model transgenic zebrafish were treated with neoglycyrrhizin (1 μg / ml) for 24 hours as the experimental group; normal transgenic zebrafish served as the normal control group; 15 juvenile zebrafish from each of the normal control group, model group, and experimental group were randomly selected. After tricaine anesthesia, the fli1 fluorescence signal of the zebrafish was collected using a fluorescence microscope, and the blood vessel diameter of the zebrafish was calculated. The results are as follows: Figure 3 As shown.

[0031] from Figure 3It can be seen that, compared with the normal control group, the blood vessel diameter of the transgenic zebrafish in the model group becomes smaller, and even part of the blood vessels are blocked and necrotic; compared with the model group, the blood vessel diameter of the transgenic zebrafish in the experimental group is significantly increased, indicating that after the intervention of neoliquiritigenin, the blood vessel condition can be significantly improved, the blood vessel diameter is increased, and neoliquiritigenin has a vascular protection effect.

[0032] Example 2:

[0033] This example relates to the experiment of the improvement effect of neoliquiritigenin on the neurological damage of middle artery embolism mice, specifically:

[0034] 1. Establishing middle artery embolism model mice

[0035] A total of 40 male C57BL / 6J mice were randomly divided into a normal control group, a model group and two experimental groups (10 mg / kg and 20 mg / kg). The normal control group was not treated. The treatment method of the model group was as follows: the mice were weighed, anesthetized with isoflurane, and fixed in a supine position on the operating table. The left common carotid artery and the initial part of the external carotid artery were exposed in turn under a stereoscopic surgical microscope. Then the proximal end of the common carotid artery was ligated, a ligation line was placed near the bifurcation of the common carotid artery, and a micro-artery clamp was placed. A small opening was cut between the two lines, and a wire plug was quickly inserted. When the wire plug was inserted 1 cm, there was a slight resistance, and the insertion was stopped immediately. At this time, the head end of the wire was just in the middle cerebral artery. The silk thread at the bifurcation was ligated, the excess silk thread was cut off, and the mouse was placed in a 37°C incubator. After 1 hour of ischemia, the wire plug was removed, the soft tissue and skin were sutured, and the middle artery embolism model was prepared. The treatment method of the two experimental groups was as follows: first, the mice were treated with neoliquiritigenin by gavage, and the dosage of neoliquiritigenin was 10 mg / kg or 20 mg / kg (10 mg or 20 mg of neoliquiritigenin per kg of mouse body weight). Half an hour later, the middle artery embolism operation was performed according to the above method.

[0036] The brain infarction volume of the model group and the experimental group mice was detected 24 hours after ischemia-reperfusion.

[0037] 2. Brain infarction volume determination

[0038] 2,3,5-chlorinated triphenyl tetrazolium (TTC) can react with succinate dehydrogenase in the mitochondria of living cells to generate red formazan, which is used to detect cell viability and is a commonly used reagent for evaluating cerebral ischemic injury. The mice in each group were decapitated and the removed brain tissues were quickly placed in a -20 °C refrigerator, removed after 10 min of freezing, and then coronally cut into 6 consecutive brain coronal sections at an interval of 2 mm according to the brain atlas. Then the brain slices were quickly placed in 5 ml of a solution containing 2% TTC, incubated at 37 °C for 30 min, and the brain slices were turned over every 10 min to ensure uniform staining. Normal tissues were stained rose red and infarcted tissues were stained white. The brain slices were removed and photographed with a digital camera. The differences in cerebral infarction volume between the groups were compared. The results are shown in Figure 4

[0039] As can be seen from Figure 4 , compared with the normal control group, the cerebral infarction volume in the model group of mice significantly increased after arterial embolism, and the body weight loss index significantly increased. Compared with the model group, the experimental group was given neoliquiritigenin 20 mg / kg intervention, and the cerebral infarction volume of the mice significantly decreased. It is shown that neoliquiritigenin can significantly reduce the cerebral infarction volume caused by middle cerebral artery embolism in mice.​

Claims

1. Application of neoglycyrrhizin in the preparation of products for the prevention or treatment of ischemic encephalopathy.

2. The application of the neoglycyrrhizin according to claim 1 in the preparation of products for the prevention or treatment of ischemic encephalopathy, characterized in that, The ischemic encephalopathy includes cerebral thrombosis, cerebral vascular stenosis / occlusion, and cerebral arteriosclerosis.

3. The application of the neoglycyrrhizin according to claim 1 in the preparation of products for the prevention or treatment of ischemic encephalopathy, characterized in that, The product is either a drug or food.

4. The application of the neoglycyrrhizin according to claim 3 in the preparation of products for the prevention or treatment of ischemic encephalopathy, characterized in that, The food products mentioned are functional foods or health products.

5. The use of the neoglycyrrhizin according to claim 1 in the preparation of products for the prevention or treatment of ischemic encephalopathy, characterized in that, Neoglycyrrhizin can improve behavioral scores, cerebral thrombosis and vascular diameter after ischemia, and reduce the volume of cerebral infarction.

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