CN116407557A - Pharmaceutical composition for preventing and treating cerebral arterial thrombosis and application thereof - Google Patents

Abstract

The invention discloses a pharmaceutical composition for preventing and treating hemorrhagic cerebral apoplexy and application thereof, relates to the technical field of medicines, and solves the technical problems that no special method is available for the hemorrhagic cerebral apoplexy in the prior art and the clinical prevention and treatment effects are limited. The invention relates to a pharmaceutical composition for preventing and treating cerebral arterial thrombosis, which consists of the following compounds or salts thereof: sulfoxylamine, deferiprone, furosemide, disulfiram, and sodium nitroprusside; the weight ratio of the sulfoxylamine, the deferiprone, the furosemide, the disulfiram and the sodium nitroprusside is 100-400:100-400:50-200:1-4. The pharmaceutical composition for preventing and treating hemorrhagic stroke can obviously reduce the mortality, the bleeding amount, the nerve injury and the inflammatory factor level of a mouse model of the hemorrhagic stroke, can provide a new medicine source for treating the hemorrhagic stroke, and has potential great economic and social benefits.

Description

A pharmaceutical composition for preventing and treating hemorrhagic stroke and its application

technical field

The invention relates to the technical field of medicine, in particular to a pharmaceutical composition for preventing and treating hemorrhagic stroke and its application.

Background technique

Stroke, commonly known as cerebrovascular accident, is an acute cerebrovascular disease, including ischemic and hemorrhagic subtypes.

Hemorrhagic stroke is a disease with acute onset and cerebral blood circulation disorder caused by cerebrovascular hemorrhage. Cerebrovascular hemorrhage in patients with hemorrhagic stroke is mainly caused by stenosis, occlusion or rupture of intracerebral arteries, clinically manifested as symptoms and signs of one-time or permanent brain dysfunction. Hemorrhagic stroke is a serious threat to human health around the world due to its high morbidity, disability, and mortality. At present, there is no effective treatment for this serious disease clinically, and it is urgent to develop new treatment methods.

Contents of the invention

The invention discloses a pharmaceutical composition for preventing and treating hemorrhagic cerebral apoplexy and its application, which solves the technical problems in the prior art that there is no special effective method for hemorrhagic cerebral apoplexy and the clinical prevention and treatment effects are limited.

In order to solve the above problems, the present invention adopts the following technical solutions:

The first aspect of the present invention provides a pharmaceutical composition for preventing and treating hemorrhagic stroke.

The pharmaceutical composition for preventing and treating hemorrhagic stroke of the present invention consists of the following compounds or salts thereof:

Buthionine sulfoxide amine, deferiprone, furosemide, disulfiram, and sodium nitroprusside;

The weight ratio of buthionine sulfoxide amine: deferiprone: furosemide: disulfiram: sodium nitroprusside is 100-400: 100-400: 100-400: 50-200: 1-4.

Preferably, the weight ratio of butthionine sulfoxide amine: deferiprone: furosemide: disulfiram: sodium nitroprusside is 100-200: 100-200: 100-200: 50-100: 1-2.

Preferably, the pharmaceutical composition for preventing and treating hemorrhagic stroke is also composed of pharmaceutically acceptable excipients or auxiliary components.

Preferably, the pharmaceutical composition for the prevention and treatment of hemorrhagic stroke contains buthionine sulfoxide amine, deferiprone, furosemide, disulfiram and sodium nitroprusside or their salts as active ingredients, adding Preparations prepared from pharmaceutically acceptable excipients or auxiliary ingredients.

The second aspect of the present invention provides the application of a pharmaceutical composition for preventing and treating hemorrhagic stroke.

Application of the pharmaceutical composition for preventing and treating hemorrhagic stroke described in any one of the technical solutions of the present invention in the preparation of medicines for preventing and treating hemorrhagic stroke.

The technical scheme adopted in the present invention can achieve the following beneficial effects:

The pharmaceutical composition for preventing and treating hemorrhagic stroke of the present invention is composed of the following compounds or salts thereof: buthionine sulfoximine, deferiprone, furosemide, disulfiram and sodium nitroprusside, wherein Thiamine sulfoximine can significantly reduce the tissue copper death in the hemorrhagic stroke model, deferiprone can significantly reduce the tissue ferroptosis in the hemorrhagic stroke model, furosemide has a significant diuretic effect on the hemorrhagic stroke model Disulfiram is an inhibitor of the porin GSDMD, which can inhibit the release of IL-1β and other DAMPs (damage associated molecular patterns, damage-related molecular patterns), and sodium nitroprusside can significantly reduce blood pressure in hemorrhagic stroke models. The combined use of buthionine sulfoxide amine, deferiprone, furosemide, disulfiram and sodium nitroprusside can significantly reduce the mortality, bleeding volume, nerve damage and inflammatory factor levels in a mouse model of hemorrhagic stroke .

On the other hand, the present invention provides the optimal ratio of each component of the drug combination, and it is found that under this ratio, it can better improve hemorrhagic stroke. Specifically, the weight ratio of butthionine sulfoxide amine: deferiprone: furosemide: disulfiram: sodium nitroprusside is 100~400: 100~400: 100~400: 50~200: 1~4, Especially preferred proportioning is: the weight ratio of buthionine sulfoxide amine: deferiprone: furosemide: disulfiram: sodium nitroprusside is 100~200: 100~200: 100~200: 50~100: 1~2.

The application of the pharmaceutical composition of the invention can provide a new drug source for the treatment of hemorrhagic cerebral apoplexy, and has potential significant economic and social benefits. The preparations prepared with buthionine sulfoxide amine, deferiprone, furosemide, disulfiram and sodium nitroprusside as active ingredients have application prospects as preventive and treatment drugs for hemorrhagic stroke, and are subject to national innovative drug approval The method is developed and developed, which is expected to become an innovative drug for the prevention and treatment of hemorrhagic stroke with high efficiency and low toxicity, and has broad prospects for industrialization.

That is, the pharmaceutical composition for preventing and treating hemorrhagic stroke of the present invention and its application solve the technical problems in the prior art that there is no specific method for treating hemorrhagic stroke and the clinical prevention and treatment effects are limited.

Detailed ways

In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Apparently, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other implementations obtained by persons of ordinary skill in the art without making creative efforts fall within the protection scope of the present invention.

Example 1

This example describes in detail the effect of the pharmaceutical composition of the present invention on the hemorrhagic stroke model.

Grouping and administration: C57BL/6 mice were randomly divided into 9 groups, namely sham operation group, model group, drug intervention group 1, drug intervention group 2, drug intervention group 3, drug intervention group 4, drug intervention group 5, Drug intervention group 6 and drug intervention group 7, 10 rats in each group. Drug intervention group 1 to drug intervention group 7 received drug combination intragastric intervention (buthionine sulfoxide amine, deferiprone, furosemide and disulfiram) and tail vein injection intervention (sodium nitroprusside), once a day, Dosing was started the day before modeling.

Modeling and sample collection: mice were anesthetized by intraperitoneal injection of 3.5% pentobarbital sodium, and fixed on the stereotaxic instrument in the prone position, fixed on the stereotaxic instrument in the prone position, local disinfection with iodine tincture and exposed skull drilling, and then used 0.3 μL of type VII collagenase (0.35 U) was injected into the micro-injector, the retention needle was retained for 5 minutes, and then the needle was pulled out slowly, the bone glue covered the skull and the skin was sutured after drilling the hole. The sham operation group was injected with the vehicle without collagenase in the same way.

Neurological function was measured 24 hours after the establishment of the hemorrhagic stroke model. The neurological function was scored using the Zea-Longa scoring standard. The more severe the hemorrhagic stroke, the more obvious the symptoms of neurological impairment, and the higher the neurological score.

4 points: The mouse has a disturbance of consciousness and cannot walk autonomously.

3 points: The body of the mouse fell to the uninjured side when walking.

2 points: the body of the mouse turns to the uninjured side in a circle, and forms a tail-chasing shape.

1 point: The uninjured forelimb of the mouse cannot be fully extended.

Measurement of the amount of cerebral hemorrhage: 24 hours after modeling, pentobarbital sodium anesthesia, PBS heart perfusion, brain tissue was taken immediately, the brain on the bleeding side was cut off, the blood on the surface of the brain tissue was washed with normal saline, and then homogenized. The supernatant was taken to detect the hemoglobin content with a hemoglobin detection kit, and the level of inflammatory factors in brain tissue was detected with an IL-1β kit, and the mortality of mice in each group was detected within 24 hours.

The drug components and weights included in drug intervention group 1 to drug intervention group 7 are as follows:

Drug intervention group 1: Buthionine sulfoximine 200 mg/kg, deferiprone 125 mg/kg, furosemide 100 mg/kg, disulfiram 50 mg/kg, sodium nitroprusside 1 mg/kg;

Drug intervention group 2: butthionine sulfoximine 100 mg/kg, deferiprone 400 mg/kg, furosemide 300 mg/kg, disulfiram 200 mg/kg, sodium nitroprusside 4 mg/kg;

Drug intervention group 3: Buthionine sulfoximine 400 mg/kg, deferiprone 100 mg/kg, furosemide 400 mg/kg, disulfiram 100 mg/kg, sodium nitroprusside 4 mg/kg;

Drug intervention group 4: Buthionine sulfoximine 400 mg/kg, deferiprone 400 mg/kg, furosemide 100 mg/kg, disulfiram 200 mg/kg, sodium nitroprusside 2 mg/kg;

Drug intervention group 5: butthionine sulfoximine 50 mg/kg, deferiprone 400 mg/kg, furosemide 300 mg/kg, disulfiram 200 mg/kg, sodium nitroprusside 4 mg/kg;

Drug intervention group 6: Buthionine sulfoximine 400 mg/kg, deferiprone 50 mg/kg, furosemide 400 mg/kg, disulfiram 100 mg/kg, sodium nitroprusside 4 mg/kg;

Drug intervention group 7: Buthionine sulfoximine 400 mg/kg, deferiprone 400 mg/kg, furosemide 50 mg/kg, disulfiram 200 mg/kg, and sodium nitroprusside 2 mg/kg.

Table 1 Effect of drug combination on mortality in hemorrhagic stroke model

group mortality rate(%) mock surgical group 0 model group 40 Drug Intervention Group 1 0 Drug Intervention Group 2 10 Drug Intervention Group 3 10 Drug Intervention Group 4 10 Drug Intervention Group 5 20 Drug Intervention Group 6 20 Drug Intervention Group 7 20

Table 1 shows the effect of the pharmaceutical composition on the mortality rate of the hemorrhagic stroke model. From the data in Table 1, it can be seen that the pharmaceutical composition used in drug intervention group 1 to drug intervention group 7 can significantly reduce the mortality rate of hemorrhagic stroke, especially the drug composition used in drug intervention group 1 has the best effect . Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

Table 2 Effect of drug combination on blood loss in hemorrhagic stroke model

group Brain tissue hemoglobin content (μg/mL) mock surgical group 232 ± 32 model group 980 ± 81 Drug Intervention Group 1 311 ± 31 Drug Intervention Group 2 370 ± 33 Drug Intervention Group 3 378 ± 36 Drug Intervention Group 4 367 ± 39 Drug Intervention Group 5 454 ± 38 Drug Intervention Group 6 483 ± 48 Drug Intervention Group 7 421 ± 40

Table 2 shows the effect of the pharmaceutical composition on the bleeding volume of the hemorrhagic stroke model. From the data in Table 2, it can be seen that the pharmaceutical composition used in drug intervention group 1 to drug intervention group 7 can significantly reduce the amount of bleeding in the hemorrhagic stroke model, especially the pharmaceutical composition used in drug intervention group 1 has the best effect. excellent. Specifically, the amount of bleeding in the model group increased by 748 μg/mL, and decreased by 89% after being treated with the composition of drug intervention group 1. Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

Table 3 Effects of drug combinations on nerve damage in hemorrhagic stroke models

group Neurological score (Zea-Longa score) mock surgical group 0 model group 2.9 ± 0.4 Drug Intervention Group 1 0.8 ± 0.2 Drug Intervention Group 2 1.2 ± 0.2 Drug Intervention Group 3 1.3 ± 0.3 Drug Intervention Group 4 1.2 ± 0.1 Drug Intervention Group 5 1.5 ± 0.3 Drug Intervention Group 6 1.6±0.4 Drug Intervention Group 7 1.6±0.3

Table 3 shows the effect of the pharmaceutical composition on nerve damage in the hemorrhagic stroke model. From the data in Table 3, it can be known that the pharmaceutical composition used in drug intervention group 1 to drug intervention group 7 can reduce the nerve damage of hemorrhagic stroke, especially the drug composition used in drug intervention group 1 has the best effect. Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

Table 4 Effect of drug combination on serum inflammatory factors in hemorrhagic stroke model

group Serum IL-1β level (ng/mL) mock surgical group 0.21 ± 0.06 model group 2.87±0.25 Drug Intervention Group 1 0.31±0.07 Drug Intervention Group 2 0.57 ± 0.06 Drug Intervention Group 3 0.62±0.05 Drug Intervention Group 4 0.68±0.07 Drug Intervention Group 5 1.17±0.12 Drug Intervention Group 6 1.23±0.10 Drug Intervention Group 7 1.15±0.11

Table 4 shows the effect of the pharmaceutical composition on the serum inflammatory factors of the hemorrhagic stroke model. From the data in Table 4, it can be seen that the pharmaceutical composition used in drug intervention group 1 to drug intervention group 7 can reduce the level of inflammatory factors in the brain tissue of the hemorrhagic stroke model, especially the pharmaceutical composition used in drug intervention group 1 Best results. Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

From the above data, it was found that the combination drugs butthionine sulfoximine, deferiprone, furosemide, disulfiram and sodium nitroprusside could significantly reduce the mortality, bleeding volume, neurological damage and inflammation in the hemorrhagic stroke model. Sex factor secretion, has excellent effect of prevention and treatment of hemorrhagic stroke; the above data also confirmed that the optimal ratio of buthionine sulfoxide amine, deferiprone, furosemide, disulfiram and sodium nitroprusside is: The weight ratio of thiamine sulfoxide amine: deferiprone: furosemide: disulfiram: sodium nitroprusside is 100~400: 100~400: 100~400: 50~200: 1~4, especially preferred The ratio is: the weight ratio of buthionine sulfoxide amine: deferiprone: furosemide: disulfiram: sodium nitroprusside is 100-200: 100-200: 100-200: 50-100: 1-2.

Example 2

This example describes in detail the drug toxicity evaluation of the pharmaceutical composition of the present invention.

Grouping and administration: C57BL/6 mice were randomly divided into 3 groups, namely sham operation group, model group, and drug combination group 1 (buthionine sulfoximine 200 mg/kg; deferiprone 125 mg/kg; Furosemide 100 mg/kg; disulfiram 50 mg/kg; sodium nitroprusside 1 mg/kg) and drug combination group 2 (buthionine sulfoxide amine 200 mg/kg; deferiprone 125 mg/kg; Disulfiram 50 mg/kg), 10 rats in each group. Drug combination group 1 and drug combination group 2 received drug combination intragastric intervention (drug combination group 1: buthionine sulfoximine, deferiprone, furosemide and disulfiram; drug combination group 2: buthionine Sulfoxide amine, deferiprone, disulfiram) and tail vein injection intervention (drug combination group 1: sodium nitroprusside; drug combination group 2: none), once a day, starting the day before modeling.

Modeling and sample collection: mice were anesthetized by intraperitoneal injection of 3.5% pentobarbital sodium, and fixed on the stereotaxic instrument in the prone position, fixed on the stereotaxic instrument in the prone position, local disinfection with iodine tincture and exposed skull drilling, and then used 0.3 μL of type VII collagenase (0.35 U) was injected into the micro-injector, the retention needle was retained for 5 minutes, and then the needle was pulled out slowly, the bone glue covered the skull and the skin was sutured after drilling the hole. The sham operation group was injected with the vehicle without collagenase in the same way.

Neurological function was measured 24 hours after the establishment of the hemorrhagic stroke model. The neurological function was scored using the Zea-Longa scoring standard. The more severe the hemorrhagic stroke, the more obvious the symptoms of neurological impairment, and the higher the neurological score.

4 points: The mouse has a disturbance of consciousness and cannot walk autonomously.

3 points: The body of the mouse fell to the uninjured side when walking.

2 points: the body of the mouse turns to the uninjured side in a circle, and forms a tail-chasing shape.

1 point: The uninjured forelimb of the mouse cannot be fully extended.

Measurement of the amount of cerebral hemorrhage: 24 hours after modeling, pentobarbital sodium anesthesia, PBS heart perfusion, brain tissue was taken immediately, the brain on the bleeding side was cut off, the blood on the surface of the brain tissue was washed with normal saline, and then homogenized. The supernatant was taken to detect the hemoglobin content with a hemoglobin detection kit, and the level of inflammatory factors in brain tissue was detected with an IL-1β kit, and the mortality of mice in each group was detected within 24 hours.

Table 5 Effect of drug combination group 1 and drug combination group 2 on the mortality rate of hemorrhagic stroke model

group mortality rate(%) mock surgical group 0 model group 40 drug combination group 1 0 drug combination group 2 10

Table 5 shows the effect of drug combination group 1 and drug combination group 2 on the mortality rate of the hemorrhagic stroke model. From the data in Table 5, it can be seen that the pharmaceutical compositions used in drug combination group 1 and drug combination group 2 can significantly reduce the mortality rate of hemorrhagic stroke, especially the effect of the drug combination group 1 is better . Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

Table 6 Effects of drug combination group 1 and drug combination group 2 on bleeding volume in hemorrhagic stroke model

group Brain tissue hemoglobin content (μg/mL) mock surgical group 232 ± 32 model group 980 ± 81 Drug Combination Group 1 311 ± 31 drug combination group 2 458 ± 42

Table 6 shows the effect of drug combination group 1 and drug combination group 2 on the bleeding volume of the hemorrhagic stroke model. From the data in Table 6, it can be seen that the pharmaceutical composition used in the drug combination group 1 and the drug combination group 2 can significantly reduce the amount of bleeding in the hemorrhagic stroke model, especially the effect of the pharmaceutical composition used in the drug intervention group 1 is the highest. excellent. Specifically, the amount of bleeding in the model group increased by 748 μg/mL, and decreased by 89% after being treated with the composition of drug intervention group 1. Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

Table 7 Effects of drug combination group 1 and drug combination group 2 on nerve damage in hemorrhagic stroke model

group Neurological score (Zea-Longa score) mock surgical group 0 model group 2.9 ± 0.4 drug combination group 1 0.8 ± 0.2 drug combination group 2 1.2 ± 0.2

Table 7 shows the effect of the pharmaceutical composition on nerve damage in the hemorrhagic stroke model. From the data in Table 7, it can be known that the pharmaceutical compositions used in drug combination group 1 and drug combination group 2 can reduce the nerve damage in hemorrhagic stroke, especially the drug composition used in drug intervention group 1 has the best effect. Values are expressed as mean ± SED. n=10, p<0.05 vs Model.

From the above data, it was found that, compared with the combination drugs buthionine sulfoxide amine, deferiprone and disulfiram, the combination drugs butthionine sulfoxide amine, deferiprone, furosemide, disulfiram and nitroprusside Sodium significantly reduces mortality, blood loss, and neurological damage in a model of hemorrhagic stroke. Specifically, disulfiram is an inhibitor of the porin GSDMD. Excessive use of disulfiram in hemorrhagic stroke has off-target effects and may cause toxic side effects, while sodium nitroprusside is a vasodilator drug and furosemide is a diuretic drug, which can promote Butthionine sulfoxide amine, which chelates copper ions, and deferiprone, which chelates iron ions, are quickly eliminated from the body and play a role in removing toxins from the body. Therefore, the combined use of sodium nitroprusside and furosemide can reduce toxins in the body and increase bleeding. The survival rate of stroke, reduce the amount of cerebral hemorrhage and nerve damage.

The above is only a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Anyone skilled in the art can easily think of changes or substitutions within the technical scope disclosed in the present invention. Should be covered within the protection scope of the present invention.

Claims (5)

1. A pharmaceutical composition for preventing and treating cerebral arterial thrombosis, which is characterized by comprising a compound or salt thereof as shown in the following formula:

sulfoxylamine, deferiprone, furosemide, disulfiram, and sodium nitroprusside;

the weight ratio of the sulfoxylamine, the deferiprone, the furosemide, the disulfiram and the sodium nitroprusside is 100-400:100-400:50-200:1-4.

2. The pharmaceutical composition for preventing and treating cerebral arterial thrombosis according to claim 1, wherein the weight ratio of the sulfoxylamine-busulfan, the deferiprone, the furosemide, the disulfiram and the sodium nitroprusside is 100-200:100-200:50-100:1-2.

3. The pharmaceutical composition for preventing and treating cerebral arterial thrombosis according to claim 1 or 2, characterized in that it also consists of pharmaceutically acceptable auxiliary materials or auxiliary components.

4. The pharmaceutical composition for preventing and treating cerebral arterial thrombosis according to claim 3, which is a preparation prepared by adding pharmaceutically acceptable auxiliary materials or auxiliary components to active ingredients such as sulfoxylamine, deferiprone, furosemide, disulfiram and sodium nitroprusside or salts thereof.

5. Use of the pharmaceutical composition for preventing and treating hemorrhagic stroke according to any one of claims 1 to 4 in the preparation of a medicament for preventing and treating hemorrhagic stroke.