WO2023024332A1 - Application of composition in preparation of drug for preventing and treating stroke - Google Patents

Abstract

An application of a composition in the preparation of a drug for preventing and treating stroke. The composition is composed of ginseng, aconite, lingzhi, safflower, artificial deer musk, bear bile, pearl, bufotoxin, borneol, and calculus bovis. The described composition can significantly improve nerve function impairment, reduce the degree of cerebral edema, and reduce the area of cerebral infarction.

Description

[Title of the invention established by ISA under Rule 37.2] Use of a composition for the preparation of a medicament for the prevention and treatment of stroke technical field

The invention relates to the field of traditional Chinese medicine, in particular to a pharmaceutical composition for preventing and treating stroke.

Background technique

Stroke is a common disease in neurology. It is often caused by the sudden rupture of blood vessels in the brain or the inability of blood to flow into the brain due to vascular obstruction, which causes brain tissue damage. It has the characteristics of high morbidity, high mortality and high disability rate. A factor of death. In recent years, due to the increasing pressure of life and work, the incidence of stroke has been increasing year by year and tends to be younger. Stroke includes ischemic stroke and hemorrhagic stroke, and the incidence of ischemic stroke is relatively high, accounting for about 75% to 90% of the total number of strokes. Ischemic stroke usually occurs in a quiet state, manifested as dysphagia, slurred articulation, crooked corners of the mouth, dysphagia, hemiplegia, sensory disturbance, and in severe cases, fecal incontinence, etc.; hemorrhagic stroke mostly occurs in an active state, It manifests as headache, nausea, jet-like vomiting, limb hemiplegia, sensory disturbance, and in severe cases, disturbance of consciousness and coma may occur.

The mechanism of brain injury caused by stroke is relatively complex, which is the result of the interaction of complex pathophysiological processes such as excitotoxicity, oxidative stress, inflammation, and apoptosis, so the efficacy of a single drug is limited.

Cerebral stroke belongs to the category of "stroke" in traditional Chinese medicine. Due to the deficiency of righteousness, diet, emotion, fatigue and internal injuries, etc., the qi and blood are reversed, resulting in wind, fire, phlegm, and blood stasis, resulting in cerebral arteries blockage or blood spillage outside the cerebral arteries. It is the basic pathogenesis, and the main clinical manifestations are sudden fainting, hemiplegia, crooked tongue, slurred or silent speech, and numbness in one side of the body. The use of traditional Chinese medicine compound can play a role in multiple links of stroke and play a good therapeutic role.

The chemical drugs currently used to treat stroke are mainly thrombolytic drugs, anticoagulant drugs and neuroprotective agents. Among them, thrombolytic drugs are used to shrink thrombus and reduce the degree of vascular occlusion. Anticoagulant drugs are mainly used to improve microcirculation and restore blood flow. Neuroprotective agents are mainly used to maintain nerves and delay nerve damage. Traditional Chinese medicine is also widely used in the treatment of stroke. Single herbs and compound drugs are mainly used to promote blood circulation and remove blood stasis, such as Angelica, Sanqi, Chuanqiong, etc.; compound drugs mainly include Compound Danshen Tablets, Buyang Huanwu Decoction, Oral drugs such as Guanxin Shutong Capsules, Yiqi Qiangxin Capsules, Bushen Yiqi Capsules, and Naoxintong Capsules, as well as injections such as Danhong Injection and Compound Danshen Injection. However, these pharmaceutical compositions often have single efficacy, complex composition, or poor safety, and lack strict and systematic pharmacodynamic experiments that meet the requirements of modern pharmacology, making it difficult to carry out strict quality control and standardized production.

Contents of the invention

Based on this, the object of the present invention is to provide a composition for preparing a medicament for preventing and treating stroke.

CN108057066B discloses a Chinese medicine composition for preventing and treating unstable angina pectoris, which consists of ginseng, aconite, ganoderma lucidum, safflower, artificial musk, bear bile, pearl, toad venom, borneol and bezoar (due to less natural sources of bezoar, generally used in vitro cultivation of bezoar). The inventor surprisingly found that the traditional Chinese medicine composition can not only relieve angina pectoris, but also has a good therapeutic effect on stroke, especially ischemic stroke. The reasonable compatibility of traditional Chinese medicine components can effectively improve neurological deficits, reduce the size of cerebral infarction, reduce the degree of brain tissue edema, resist oxidative stress damage, relieve the symptoms of stroke from various angles, and block the progress of the disease; and it is safe and reliable , No toxic side effects on the human body.

The specific technical scheme is as follows:

An application of a composition in the preparation of a medicament for preventing and treating cerebral apoplexy, the composition mainly consists of the following components by weight:

In some embodiments of the present invention, the composition mainly consists of the following components by weight:

In some embodiments of the present invention, the composition mainly consists of the following components by weight:

In some embodiments of the present invention, the composition mainly consists of the following components by weight:

In some embodiments of the present invention, the medicament is prepared from the composition and pharmaceutically acceptable excipients.

In some embodiments of the present invention, the drug can be prepared according to conventional methods in the art. The medicament of the present invention is a composition obtained by pulverizing and mixing raw materials, or an extract obtained by mixing raw materials, or an extract obtained by separately extracting raw materials and mixing them. Optionally, the extract is further refined and purified. Wherein, the extraction method includes decoction extraction, reflux extraction, immersion extraction, ultrasonic extraction, percolation extraction, microwave extraction, etc.; the purification method includes water extraction and alcohol precipitation, alkali-soluble acid precipitation and various column chromatography purification methods, Such as macroporous resin column, silica gel column, gel column, reversed phase column, etc.

In some embodiments of the present invention, the dosage form of the drug is tablet, capsule, granule, pill, oral liquid or drop pill. The above composition is added with suitable pharmaceutically acceptable auxiliary materials, and can be made into corresponding tablets, capsules, granules, pills, oral liquids or drop pills according to the conventional preparation process of the required dosage form.

The pharmaceutically acceptable adjuvant refers to the conventional pharmaceutical adjuvant in the field of pharmacy, selected from fillers, binders, disintegrants, lubricants, suspending agents, wetting agents, pigments, essences, solvents, surfactants Or one or more flavoring agents. The filler is selected from starch, sucrose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose or glucose, etc.; the binder is selected from cellulose derivatives, alginate, starch, water, Dextrin, gelatin or polyvinylpyrrolidone, etc.; the disintegrant is selected from microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose or cross-linked sodium carboxymethyl cellulose ; Described lubricant is selected from stearic acid, macrogol, calcium carbonate, sodium bicarbonate, micropowder silica gel, talcum powder or magnesium stearate; Described suspending agent is selected from micropowder silica gel, beeswax, cellulose, solid polyethylene glycol; the wetting agent is selected from glycerin, Tween-80, ethoxylated hydrogenated castor oil or lecithin; the solvent is selected from ethanol, liquid polyethylene glycol, isopropanol, Tween-80 , glycerin, propylene glycol or vegetable oil, and the vegetable oil is selected from soybean oil, castor oil, peanut oil, blended oil, etc.; the surfactant is selected from sodium dodecylbenzenesulfonate, stearic acid, polyoxyethylene-polyoxy Propylene copolymer, fatty acid sorbitan or polysorbate (Tween), etc.; the flavoring agent is selected from aspartame, sucralose, essence, stevia, acesulfame potassium, citric acid or sodium saccharin.

In some embodiments of the present invention, the composition and the medicine prepared therefrom are used for preventing and treating stroke, and the stroke is ischemic stroke.

In some embodiments of the present invention, the composition and the medicine prepared therefrom improve neurological deficits, and/or reduce the size of cerebral infarction, and/or reduce the degree of brain tissue edema, and/or resist oxidative stress damage. In some embodiments of the present invention, the anti-oxidative stress damage is to reduce MDA content, increase GSH content and SOD activity (P<0.05).

Beneficial effects of the present invention:

The composition of the present invention can effectively improve neurological deficits, reduce the size of cerebral infarction, reduce the degree of brain tissue edema, resist oxidative stress damage, and relieve the symptoms of stroke from various angles through the reasonable compatibility of various traditional Chinese medicine components. Block the progress of the disease; controllable quality, easy to standardize production; and safe and reliable, non-toxic and side effects on the human body.

Detailed ways

The following are specific embodiments of the present invention, and the described examples are to further describe the present invention, but do not limit the present invention.

Example 1

The test drug of this embodiment experimental group is made up of the following components by weight:

Example 2

The test drug of this embodiment experimental group is made up of the following components by weight:

Example 3

The test drug of this embodiment experimental group is made up of the following components by weight:

Example 4

The test drug of this embodiment experimental group is made up of the following components by weight:

Example 5

The test drug of this embodiment experimental group is made up of the following components by weight:

Example 6

The test drug of this embodiment experimental group is made up of the following components by weight:

Pharmacodynamic study: effects on cerebral ischemia-reperfusion model rats

1. Experimental method

1.1 Animal grouping and drug intervention

Get 160 male SD rats (body weight 240 ± 20g), after adaptive feeding 1～2, be divided into 8 groups at random, be respectively sham operation group, model group and embodiment 1,2,3,4,5, 6 groups (wherein, the medicinal taste of each group of embodiment is pulverized according to the prescription quantity, sieved and mixed, suspended with sodium carboxymethylcellulose solution to make a suspension, shaken up before administration), totally 8 groups, each group of 20 Only, begin to give corresponding drug gavage every day (embodiment 1,2,3,4,5,6 groups are administered by 57,63,67,71,83,89mg crude drug amount/Kg body weight respectively, sham operation group, model group was given an equal amount of sodium carboxymethylcellulose solution). Are administered for 4 weeks. Modeling was established on the second day after the end of administration.

1.2 Preparation of cerebral ischemia-reperfusion model rats

12h fasting before operation, free drinking water. Rats in each group were anesthetized by intraperitoneal injection of chloral hydrate, fixed on the operating table in lateral position, and the anal temperature of rats was maintained at 37±1°C during the experiment. The operation was performed in the following steps: skin preparation, skin disinfection; midline incision in the neck, exposure of the common carotid artery, blunt separation of the internal carotid artery, clamping of the common carotid artery and the internal carotid artery, and slow insertion of the nylon thread through the main incision of the external carotid artery. Gently advance the internal carotid artery into the cranium, mark the bifurcation of the common carotid artery, encounter slight resistance when advancing about 18mm, and then reach the thinner middle cerebral artery to stop, ligate and block the middle cerebral artery, and make the local cerebral blood flow sufficient The wound was blocked and sutured, and a nylon thread about 3 cm in length was placed outside the body, and the operation area was disinfected with povidone iodine. After 1 hour of ischemia, the occlusion thread was pulled out for about 10 minutes to achieve reperfusion. Sham-operated group: No insertion and ligation, and the rest of the treatment was the same as that of the drug-administered group. During the operation, the cerebral blood flow was monitored by transcranial Doppler flow analyzer.

1.3 Measuring indicators

1.3.1 Behavior defect: within 24 hours of ischemia-reperfusion, closely observe the animals in each group, and evaluate the behavior defect with reference to the Zea Longa scoring standard, where 0 is normal without nerve damage symptoms; 1 is unable to fully extend the contralateral forelimb; 2 Divided into circling to the opposite side when crawling; 3 divided into body dumping to the opposite side when walking; 4 divided into inability to walk spontaneously and loss of consciousness.

1.3.2 Cerebral infarction volume and brain water content: After administration and reperfusion for 24 hours, the animals in each group were sacrificed, and the whole brains of 10 rats were taken, and the wet weight of the brain tissue was weighed, sliced, 2% triphenyl chloride Stain in tetrazolium (TTC) solution for 30 min, fix, separate pale area (infarct area, not stained) and non-pale area (normal area), cerebral infarction volume (%) = infarct area wet weight/brain tissue wet weight ×100%. Dry the stained brain tissue in an oven to constant weight, weigh its dry weight, and calculate the water content of the brain according to the following formula: brain water content (%)=(wet weight of brain tissue-dry weight of brain tissue)/wet weight of brain tissue ×100%.

1.3.3 Oxidative stress indicators: 24 hours after administration, the animals in each group were sacrificed, and 10 rats were taken, and the brain was quickly weighed, and a 10% brain tissue homogenate was prepared with normal saline. Centrifuge at low temperature, take the supernatant, and detect the content of malondialdehyde (MDA), reduced glutathione (GSH) and superoxide dismutase (SOD) activity in the brain tissue strictly according to the kit instructions.

2. Experimental results

2.1 Comparison of behavioral deficit scores of rats in each group

Compared with the sham operation group, the behavioral scores of rats in the model group were significantly higher. Compared with the model group, the behavioral score of the rats given the Example drug decreased significantly (P<0.05), indicating that the Example drug can improve neurological deficits. The results are shown in Table 1.

Table 1 Comparison of behavioral deficit scores of rats in each group (n=20)

group	Behavioral score (points)
mock surgical group	0
model group	3.40±0.55
Example 1 group	2.61±0.44*
Example 2 group	2.33±0.29*
Example 3 groups	2.47±0.27*
Embodiment 4 groups	2.63±0.38*
Embodiment 5 groups	2.52±0.31*
Embodiment 6 groups	2.35±0.45*

Note: Compared with the model group, *P<0.05

2.2 Comparison of cerebral infarction volume and brain water content in rats in each group

There was no infarction in the brain tissue of the rats in the sham operation group; compared with the sham operation group, the brain water content of the rats in the model group was significantly higher (P<0.05). Compared with the model group, the cerebral infarction volume and brain water content of the rats given the Example drug were significantly decreased (P<0.05), indicating that the Example drug can reduce the cerebral infarction volume and brain water content caused by ischemia. The results are shown in Table 2.

Table 2 Comparison of cerebral infarction volume and brain water content in rats in each group (%, n=10)

group	infarct volume	brain water content
mock surgical group	-	66.97±4.39*
model group	24.08±1.32	78.47±6.28
Example 1 group	22.19±1.77*	74.89±3.26*

Example 2 group	22.08±1.65*	73.29±6.17*
Example 3 groups	21.63±1.28*	71.32±4.20*
Embodiment 4 groups	21.49±2.09*	71.56±6.24*
Embodiment 5 groups	21.86±2.60*	73.20±5.17*
Embodiment 6 groups	21.60±1.83*	72.02±3.19*

Note: Compared with the model group, *P<0.05

2.3 Comparison of oxidative stress indicators in rats in each group

Compared with the sham operation group, the MDA content in the brain tissue of the rats in the model group was significantly higher, while the GSH content and SOD activity were significantly lower (P<0.05). Compared with the model group, the MDA content of the rats given the example drug was significantly reduced, and the GSH content and SOD activity were significantly increased (P<0.05), indicating that the example drug can reduce oxidative stress damage. The results are shown in Table 3.

Table 3 Comparison of oxidative stress indicators in rats in each group (n=10)

group	MDA (μmol/mg Prot)	GSH (μg/mg Prot)	SOD (μU/mg Prot)
mock surgical group	9.13±0.76*	46.84±6.92*	69.05±8.14*
model group	56.30±7.18	27.80±3.21	30.43±4.85
Example 1 group	45.51±3.92*	33.41±3.35*	41.74±6.32*
Example 2 group	41.42±4.55*	35.50±5.24*	46.21±4.49*
Example 3 groups	39.21±3.26*	36.45±4.06*	45.35±6.12*
Embodiment 4 groups	40.25±5.47*	36.37±2.81*	38.72±4.50*
Embodiment 5 groups	43.07±4.84*	34.01±5.65*	36.19±5.33*
Embodiment 6 groups	39.84±3.27*	33.79±3.34*	45.84±3.26*

Note: Compared with the model group, *P<0.05

3. Experimental conclusion

The risk factors of stroke (hypertension, hyperglycemia, etc.) and ischemia will lead to the activation of reactive oxygen species (ROS)-related enzyme systems, producing a large amount of ROS, which far exceeds the endogenous scavenging capacity of the body's antioxidant defense system, and oxidative stress occurs A large number of studies have shown that compared with normal people, SOD, GSH, catalase (CAT), glutathione peroxidase (GSH-Px), and total antioxidant capacity (TAC) in stroke patients are significantly higher after the onset of stroke. decreased, MDA was significantly increased, and changes in the above indicators aggravated the disease by promoting platelet aggregation, increasing endothelial permeability, and focal lesions of endothelial cells.

In this study, the rat focal cerebral ischemia-reperfusion model was prepared by suture method. Compared with the model group, the behavioral score, cerebral infarction volume, brain water content and MDA content in the brain tissue of the drug group in the example were significantly reduced. , GSH content and SOD activity in the brain tissue were significantly increased (P<0.05), indicating that the drug of the embodiment can improve neurological impairment, reduce the degree of cerebral edema, and reduce the size of cerebral infarction. This effect may be related to the reduction of oxidative stress, It is related to the protection of brain tissue from oxygen free radical damage. The pharmaceutical composition of the present invention relieves the symptoms of cerebral apoplexy from various angles and blocks the progression of the disease through the reasonable compatibility of various traditional Chinese medicine components; it is safe and reliable, and has no toxic and side effects on the human body.

In summary, the pharmaceutical composition of the present invention can significantly reduce oxidative stress damage in ischemic stroke model animals, improve neurological deficits, protect brain tissue, and is suitable for the prevention and treatment of ischemic stroke.

The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, should be considered as within the scope of this specification.

The above-mentioned embodiments only express several implementation modes of the present invention, and the descriptions thereof are relatively specific and detailed, but should not be construed as limiting the patent scope of the invention. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

Claims (10)

1. An application of a composition in the preparation of a medicament for preventing and treating cerebral apoplexy, the composition mainly consists of the following components by weight:

   
2. The application of the composition according to claim 1 in the preparation of a medicament for the prevention and treatment of cerebral apoplexy, wherein the composition is mainly composed of the following components by weight:

   
3. The application of the composition according to claim 2 in the preparation of a medicament for the prevention and treatment of cerebral apoplexy, wherein the composition is mainly composed of the following components by weight:

   

   
4. The application of the composition according to claim 3 in the preparation of a medicament for the prevention and treatment of cerebral apoplexy, wherein the composition is mainly composed of the following components by weight:

   
5. The use of the composition according to any one of claims 1-4 in the preparation of medicaments for the prevention and treatment of cerebral apoplexy, characterized in that the bezoar used in the composition is in vitro cultured bezoar.
6. According to the application of the composition according to any one of claims 1-4 in the preparation of a medicament for the prevention and treatment of stroke, it is characterized in that the medicine is composed of the composition and pharmacy according to any one of claims 1-4 Prepared with the above acceptable excipients.
7. The application of the composition according to claim 6 in the preparation of a medicament for preventing and treating cerebral apoplexy, wherein the dosage form of the medicament is tablet, capsule, granule, pill, oral liquid or drop pill.
8. The application of the composition according to claims 1-4 in the preparation of medicines for the prevention and treatment of cerebral apoplexy, characterized in that, the medicine is a composition obtained by mixing the raw materials after pulverization; or after mixing the raw materials The extract obtained by extraction, or the extract obtained by mixing each API after extraction alone; optionally, the extract is further refined and purified.
9. The use of the composition according to any one of claims 1-4 in the preparation of a medicament for the prevention and treatment of stroke, characterized in that the stroke is ischemic stroke.
10. The application of the composition according to any one of claims 1-4 in the preparation of a medicament for the prevention and treatment of cerebral apoplexy, characterized in that the medicament improves neurological deficits, and/or reduces the size of cerebral infarction, and/or Reduce brain tissue edema, and/or protect against oxidative stress damage.