KR20210058592A - A pharmaceutical composition for treating cognitive decline due to stroke containing sildenafil and Rho-associasted kinase inhibitor - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating cognitive decline due to stroke, including sildenafil and GSK429286A. The present inventors have performed research, based on clinical knowledge related to stroke treatment, in order to use sildenafil and a ROCK inhibitor in combination to treat sequelae of stroke, and as a result, have confirmed for the first time that cognitive decline caused by stroke is significantly alleviated when sildenafil and the ROCK inhibitor are administered in combination compared to when sildenafil or the ROCK inhibitor are administered alone. Thus, it is expected that a pharmaceutical composition according to the present invention will be advantageously used for treating or studying cognitive decline-associated sequelae caused by stroke and for studying an improvement in cognitive functions.

Description

A pharmaceutical composition for treating cognitive decline due to stroke containing sildenafil and Rho-associasted kinase inhibitor}

The present invention relates to a pharmaceutical composition for the treatment of sequelae due to ischemic cerebrovascular disease comprising a phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor) and a ROCK inhibitor (Rho-associasted kinase inhibitor), and more specifically It relates to a pharmaceutical composition for treating cognitive decline due to stroke, comprising sildenafil and GSK429286A.

Stroke is defined as a condition in which consciousness disorders and neurological symptoms appear rapidly due to cerebrovascular disorders such as cerebral hemorrhage and cerebral infarction. It is reported that there are 5,000 new stroke patients.

Stroke is a temporary brain disease caused by blockage or rupture of blood vessels going to the brain.It is a disease that leaves serious sequelae, and bleeding such as loss of motor function, sensory abnormality, cognitive, speech impairment, loss of consciousness, impaired swallowing function due to damage to cranial nerve cells And various neurological symptoms depending on the site of the occlusion.

In addition, causes of cranial nerve cell damage caused by stroke include excessive excitatory neurotransmitter release, generation of free radicals, inhibition of protein synthesis, abnormal gene expression and activation of immune responses, and the complexity of the mechanism of cranial nerve cell damage. For this reason, no treatment has been developed yet to protect the brain's nerve cells from damage.

However, there are a number of studies on the use of pharmaceutical compositions containing other compounds to prevent or treat stroke sequelae (Korean Patent Publication No. 10-1869322), but phosphodiesterase type 5 activity inhibitors and ROCK inhibitors There is no research on the therapeutic use of a pharmaceutical composition containing a loss of motor function due to stroke.

The present inventors studied to use sildenafil and a ROCK inhibitor in combination in the treatment of sequelae due to stroke based on clinical knowledge related to stroke treatment. When administered, it was confirmed for the first time that the decrease in cognitive function due to stroke was significantly improved or that cognitive function was improved compared to the normal control group, and the present invention was completed based on this.

Thus, the present invention is a phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And

An object of the present invention is to provide a pharmaceutical composition for the treatment of cognitive decline caused by ischemic cerebrovascular disease, comprising a ROCK inhibitor (Rho-associasted kinase inhibitor).

In addition, the present invention is a phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And

Another object is to provide a pharmaceutical composition for enhancing cognitive function, comprising a ROCK inhibitor (Rho-associasted kinase inhibitor).

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention as described above, the present invention

Phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And

It provides a pharmaceutical composition for the treatment of cognitive decline caused by ischemic cerebrovascular disease, comprising a ROCK inhibitor (Rho-associasted kinase inhibitor).

In one embodiment of the present invention, the ischemic cerebrovascular disease may be any one selected from the group consisting of ischemic stroke (cerebral infarction), thrombosis, embolism, transient ischemic attack, leukodystrophy, and small infarction.

In another embodiment of the present invention, the phosphodiesterase type 5 activity inhibitor is mirodenafil, sildenafil, vardenafil, tadalafil, udenafil ), dasantafil, avanafil, and pharmaceutically acceptable salts, solvates, and hydrates thereof.

In another embodiment of the present invention, the ROCK inhibitor is Fasudil, Ripasudil, RKI-1447, Y-27632, GSK429286A, Y-30141, and pharmaceutically acceptable salts and solvates thereof. And it may be any one selected from the group consisting of a hydrate.

In another embodiment of the present invention, the pharmaceutical composition may be for administration after cerebrovascular occlusion, before reperfusion, simultaneously with reperfusion, or after reperfusion.

In addition, the present invention provides a method of treating cognitive decline caused by ischemic cerebrovascular disease, comprising administering the pharmaceutical composition to an individual.

In addition, the present invention provides the use of the pharmaceutical composition for the treatment of cognitive decline caused by ischemic cerebrovascular disease.

In addition, the present invention is a phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And

It provides a pharmaceutical composition for enhancing cognitive function, comprising a ROCK inhibitor (Rho-associasted kinase inhibitor).

In addition, the present invention provides a method for enhancing cognitive function, comprising administering the pharmaceutical composition to an individual.

In addition, the present invention provides the use of enhancing the cognitive function of the pharmaceutical composition.

The present inventors studied to use sildenafil and a ROCK inhibitor in combination in the treatment of sequelae due to stroke based on clinical knowledge related to stroke treatment. When administered, it was confirmed for the first time that the decrease in cognitive function due to stroke is significantly improved or that cognitive function is improved compared to the normal control group. It is expected to be useful in treatment or research and research related to cognitive function enhancement.

1 shows the escape hole used in the Barnes maze test.
Figure 2 shows the tracing figures for confirming the cognitive function of the control group, stroke (cerebral infarction) animal model, drug-only treatment group and drug combination treatment group. Here, MCAo30 represents a stroke (cerebral infarction) animal model, SIL represents a group treated with sildenafil alone, ROCK-I represents a group treated with a ROCK inhibitor (Rho-associasted kinase inhibitor) alone, and SR represents a group treated with sildenafil ) And ROCK inhibitor (Rho-associasted kinase inhibitor) are shown in combination treatment group, pre is the administration group before reperfusion after middle cerebral artery occlusion, and post is the administration group after reperfusion after middle cerebral artery occlusion.
3 is a graph showing the results of confirming the level of improvement in cognitive decline in the control group, the stroke (cerebral infarction) animal model, the drug alone treatment group, and the drug combination treatment group. Here, MCAo30 represents a stroke (cerebral infarction) animal model, SIL represents a group treated with sildenafil alone, ROCK-I represents a group treated with a ROCK inhibitor (Rho-associasted kinase inhibitor) alone, and SR represents a group treated with sildenafil ) And ROCK inhibitor (Rho-associasted kinase inhibitor) are shown in combination treatment group, pre is the administration group before reperfusion after middle cerebral artery occlusion, and post is the administration group after reperfusion after middle cerebral artery occlusion.
Figure 4 shows the results of confirming the improvement of the reduction in neuroplasticity due to the stroke by measuring the fEPSP slope in the control group, stroke (cerebral infarction) animal model, and sildenafil treatment group alone. Here, MCAo30 represents a stroke (cerebral infarction) animal model, SIL represents a group treated with sildenafil alone, pre represents a group administered before reperfusion after middle cerebral artery occlusion, and post represents a group administered after reperfusion after middle cerebral artery occlusion. Is shown.
5 is a graph showing the improvement in neuroplasticity reduction due to stroke by measuring the fEPSP slope in the control group, stroke (cerebral infarction) animal model, and sildenafil treatment group alone. Here, MCAo30 represents a stroke (cerebral infarction) animal model, SIL represents a group treated with sildenafil alone, pre represents a group administered before reperfusion after middle cerebral artery occlusion, and post represents a group administered after reperfusion after middle cerebral artery occlusion. Is shown.

Hereinafter, the present invention will be described in detail.

The present inventors studied to use sildenafil and a ROCK inhibitor in combination in the treatment of sequelae due to stroke based on clinical knowledge related to stroke treatment.As a result, sildenafil and a ROCK inhibitor were used in combination compared to the case where sildenafil and ROCK inhibitor were administered alone. In the case of administration, it was first confirmed that the decrease in cognitive function due to stroke was significantly improved or that cognitive function was improved compared to the normal control group, and the present invention was completed based on this.

Thus, the present invention is a phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And it provides a pharmaceutical composition for the treatment of cognitive decline caused by ischemic cerebrovascular disease, comprising a ROCK inhibitor (Rho-associasted kinase inhibitor).

The term "ischemic cerebrovascular disease" according to the present invention means a generic term for a disease that occurs when blood vessels in the brain are blocked for some reason and blood required for the brain is not supplied, and is also referred to as cerebral ischemic disease, and the ischemic cerebrovascular disease is brain It refers to a disease caused by the death of nerve cells in tissues known to be sensitive to cerebral ischemia, as the supply of oxygen and glucose is not performed normally due to decreased blood flow of the brain.For example, ischemic cerebrovascular disease is ischemic stroke. (Cerebral infarction), thrombosis, embolism, transient ischemic attack, leukoplakia, and small infarction, and the like, preferably ischemic stroke, but is not limited thereto.

Symptoms of "cognitive function decline due to ischemic cerebrovascular disease", which is a disease target in the present invention, are very complex and comprehensive symptoms related to perception, analysis, language, memory, and judgment, and vary depending on the location and degree of brain damage. It indicates a decrease in cognitive function, and includes, but is not limited to, a decrease in learning ability, a decrease in spatial ignorance, a decrease in spatio-temporal perception, a decrease in attention, and a decrease in memory.

In the present invention, "phosphodiesterase type 5 inhibitor (PDE inhibitor)" is an enzyme found in various tissues. The PDE inhibitors provide strong inhibition of selected alloenzymes without side effects arising from non-selective inhibitors, and PDE-5 inhibitors are known to be used in the treatment of primary pulmonary hypertension and erectile dysfunction. Examples include mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, and avanafil. And the like, and preferably sildenafil, but is not limited thereto.

In the present invention, the "ROCK inhibitor (Rho-associasted kinase inhibitor)" is a substance that functions to inhibit apoptosis, and is an action-induced Ca ^{2 in the regeneration of neurites, myosin phosphorylation and smooth muscle contraction. +} sensitizer is known that the function of such inhibition of ^{(agonist-induced Ca 2+ sensitization)} . More specifically, ROCK inhibitors are reported to alleviate the abnormal structures of muscle cells that cause hypertension and asthma, increase blood flow to the optic nerve nipple, and continuously reduce intraocular pressure. For example, Fasudil , Ripasudil, RKI-1447, Y-27632, GSK429286A, Y-30141, and the like, preferably GSK429286A (N-(6-Fluoro-1H-indazol-5-yl )-6-methyl-2-oxo-4-[4-(trifluoromethyl)phenyl]-3,4-dihydro-1H-pyridine-5-carboxamide), but is not limited thereto.

[Formula 1]

In addition, the pharmaceutical composition may be for administration after cerebrovascular occlusion, before reperfusion, simultaneously with reperfusion, or after reperfusion, preferably after middle cerebral artery occlusion, before reperfusion, simultaneously with reperfusion, or after reperfusion, but is limited thereto. In the present invention, "reperfusion" is used to resume blood vessel flow using surgery or thrombolytic agents when the supply of oxygen and nutrients to cells is cut off due to a temporary interruption of blood flow in organs or tissues. Say action.

As described above, in an embodiment of the present invention, sildenafil (phosphodiesterase type 5 activity inhibitor) and GSK429286A (ROCK inhibitor) were administered in combination to treat cognitive decline occurring in an acute stroke (cerebral infarction) animal model. In the case, it was observed that the escape hole was reached at a level similar to the escape time of the normal control group, and it was confirmed that cognitive decline was significantly improved, and some individuals in the combination administration group had a period of reaching the escape hole compared to the normal control group. By shortening, it was confirmed that it showed improved cognitive function (improvement of memory) compared to the normal control group without stroke (see Example 2).

In addition, the results of the above examples show that when sildenafil (phosphodiesterase type 5 activity inhibitor) and GSK429286A (ROCK inhibitor) were administered in combination compared to the case where sildenafil or GSK429286A was administered alone, the escape time was shortened compared to the normal control group. In comparison, it means that cognitive function is improved, suggesting that the pharmaceutical composition comprising sildenafil and GSK429286A according to the present invention can be usefully used as a cognitive function enhancer.

The pharmaceutical composition according to the present invention comprises a phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And a ROCK inhibitor (Rho-associasted kinase inhibitor) as an active ingredient, and may also include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in formulation, and includes, but is limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, and the like. It is not, and other conventional additives such as antioxidants and buffers may be further included as needed. In addition, diluents, dispersants, surfactants, binders, lubricants, and the like may be additionally added to form injection formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, it can be preferably formulated according to each component using a method disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in its formulation, but may be formulated as an injection, an inhalant, an external preparation for the skin, or an oral ingestion.

The pharmaceutical composition of the present invention can be administered orally or parenterally according to a desired method (for example, intravenous, subcutaneous, skin, nasal, airway), and the dosage is the patient's condition, weight, and disease. It depends on the degree of the drug, the form of the drug, the route of administration and the time, but may be appropriately selected by those skilled in the art.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, and drug activity of the patient. , Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

Specifically, the effective amount of the composition according to the present invention may vary depending on the age, sex, and body weight of the patient, and generally 0.001 to 150 mg, preferably 0.01 to 100 mg per 1 kg of body weight is administered daily or every other day, or 1 It can be administered in 1 to 3 times a day. However, the dose may be increased or decreased depending on the route of administration, cognitive decline due to ischemic cerebrovascular disease, preferably the severity of cognitive decline due to ischemic stroke, sex, weight, age, etc. It is not limiting.

On the other hand, as another aspect of the present invention, the present invention provides a method for controlling or treating cognitive decline according to ischemic cerebrovascular disease, comprising administering the pharmaceutical composition to an individual, preferably according to ischemic stroke. A method for controlling or treating cognitive decline is provided, but is not limited thereto.

The term "treatment" used in the present invention improves or benefits from cognitive decline due to ischemic cerebrovascular disease, preferably cognitive decline due to ischemic stroke, by administration of the pharmaceutical composition according to the present invention. It means all actions that change.

In the present invention, "individual" means a subject in need of a method for controlling or treating a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, It means mammals such as horses and cattle.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention. However, the following examples are provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Experimental materials and test methods

1-1. Experimental animals

Male rats, Sprague-Dawley, 8 weeks old and 260-280 g of experimental animals were used, and the experimental animals were obtained from Soonchunhyang University Laboratory Animal Center (Cheonan). All experimental animals were provided with commercial feed and water ad libitum under certain temperature, humidity and lighting conditions (contrast cycle 12:12, 22±2°C, 55±5%). All animal protocols were approved by the administrative panel of Soonchunhyang University Laboratory Animal Care (license number SCH16-0024). In carrying out the experiment related to the present invention, it was attempted to avoid the pain of the experimental animals and to minimize the number of experimental animals used in the experiment.

1-2. Induction of Middle Cerebral Artery occlusion (MCAo)

The rat was anesthetized with 2% isoflurane (2% isoflurane in mixed gas of oxygen and nitrous oxide, 0.3: 0.7), and the body temperature was maintained at 37 ℃ using a heating pad during surgery.

All micro-forceps were sterilized. The tip of 4-0 monofilament (Nylon suture, Medtronic plc.) is coated with dental silicone to a thickness of 0.37±0.02 mm, and the filament piece is made of 0.01% (w/v) poly-L-lysine (Sigma, USA). Coated.

The monofilament was inserted into the left common carotid artery (CCA) and proceeded toward the internal carotid artery (ICA) at 18 mm to block the origin of the middle cerebral artery (MCA).

Next, a laser Doppler probe (Millwey, Axminste, UK) was fixed to the parietal bone to measure the local cortical blood flow supplied by the middle cerebral artery during surgery. It was considered successful.

Then, transient local ischemia was induced for 30 minutes by occlusion of the left middle cerebral artery. Thirty minutes after the onset of ischemia, the rat recovered from anesthesia and the middle cerebral artery was reperfused from transient ischemia.

To verify the induction of middle cerebral artery ligation (occlusion), all rats after surgery were confirmed to be capable of the following actions:

① Cannot extend the other front leg

② Unable to circling toward contralateral to infarct after neurological deficit

The pharmaceutical composition according to the present invention was applied to the induction model of middle cerebral artery ligation (occlusion), and several behavior tests, electrophysiology, and immunohistochemistry experiments were performed in a designated time course. It was used for.

The induction model of middle cerebral artery ligation (occlusion) without experience in the behavioral test and the like and normal rats of the same age were used as a control group.

1-3. Drug treatment

The present inventors applied i) Sildenafil citrate, ii) GSK429286A and iii) Sildenafil citrate and GSK429286A to experimental animals, respectively, to direct brain injury caused by a minor ischemic stroke or a phosphodiesterase type 5 activity inhibitor (phosphodiesterase) against various phenotypic sequelae. type 5 inhibitor) and/or ROCK inhibitor (Rho-associasted kinase inhibitor).

More specifically, the following experimental drugs were administered intraperitoneally 30 minutes before reperfusion after middle cerebral artery occlusion and 30 minutes after reperfusion after middle cerebral artery occlusion.

I) Sildenafil citrate (20mg/kg/saline, Hanmi Parm. Co., Republic of Korea)

Ii) GSK429286A (10mg/kg/DMSO, Sigma, USA)

Iii) Sildenafil citrate (20mg/kg/saline) and GSK429286A (10mg/kg/DMSO)

1-4. Barnes maze test

Barnes maze consists of a circular platform (122 cm in diameter) with 20 holes around the same distance, rising 105 cm from the bottom. Four different visual cues were placed in each quadrant wall surrounding the platform at a height easy to observe the experimental animals, and one hole in the quadrant area (escape hole) contains an escape chamber under the platform. In addition, bedding was added to the escape chamber for the safety of the experimental animals. To amplify anxiety for the motivation to find the escape chamber, a metronome and bright lighting were used. More specifically, a metronome was used to continuously generate noise of 80 Hz and the illuminance measured at the center of the platform was maintained at 300 lux.

During the acclimation phase, the experimental animal was placed in a black cylinder at the center of the platform, and the acclimation phase lasted for 4 days. After 10 seconds, when the cylinder was removed, the experimental animal navigated the platform from the center and this activity was recorded by a video camera. In addition, a time of 180 seconds was given for the experimental animal to find the escape hole. When the experimental animal entered the escape chamber through the escape hole within 180 seconds, a cover was placed on the hole for 120 seconds to block the light and stop the noise of 80 Hz.

In addition, when the test animal could not find the escape hole, the test animal was carefully pulled out and guided to the escape hole with the escape chamber to induce learning about the space. The learning was an adaptation step to improve spatial memory for finding an exit, and was performed on each experimental animal for 3 times at 15 minute intervals.

In the space acquisition phase (probe phase) on the 5th day, 90 seconds were given to the experimental animals to find an escape, but the escape chamber was removed. This step was performed only once.

The distance and duration traveled in the quadrant area with the escape chamber connected to the escape hole was analyzed with Ethovision® software and the waiting time to reach the escape chamber connected to the escape hole was analyzed manually.

1-5. fEPSP slope measurement

Field potentials (fEPSP) were recorded at a slightly modified site from CA1 in the hippocampus. After the behavioral test, the animals were anesthetized intraperitoneally using urethane (1.5 g/kg) and placed in a stereotaxic frame. A temperature controller (Harvard Instruments, USA) was used to maintain the rectal temperature at 37±0.3° C. during the surgical procedure.

Next, the scalp was opened and separated, and a hole was drilled through the skull to introduce electrodes.

The coordinates (in mm) based on the bregma (the junction of the coronary artery of the skull and the sagittal suture) are as follows.

I) Recording electrode (to the Schaffer collateral): 4.0 posterior to Bregma, 3.0 laterally to centerline, 2.5 depth

Ii) Stimulating electrode (to the stratum radiatum of CA1): 3.5 posterior to Bregma, 2.0 laterally to centerline, 3.5 depth to hippocampus.

The electrode depth was finally determined by observing whether or not the induced response was optimized. fEPSP was adjusted to ~60% of the maximum response size for the test, and stimulation was generated by the BNC-2110 apparatus (National Instruments, USA) and Digital Stimulus Isolation unit (Getting Instruments, CA, USA). The response of the pyramidal neuron to the Schaffer collateral stimulation was P55 A.C. It was recorded using a pre-amplifier (3-1000 Hz bandpass, Astro-Med Inc.), and WinLTP ver. It was analyzed using 2.01 software (WinLTP Ltd.). In addition, a response occurred by a single pulse stimulation and was delivered at intervals of 20 seconds. A stable baseline was recorded for 30 to 60 minutes.

Long-term potential (LTP) is a strong theta patterned stimulus (sTPS, four trains of 10 bursts of 5 pulses at 400 Hz with a 200-ms inter-burst interval, 15-s inter-train interval). Induced by This is because sTPS (bursts of 400 Hz stimuli) are NMDA-receptor dependent and potent LTP in CA1 and DG regions is induced by sTPS in vivo.

To analyze the change in fEPSP, the slope of fEPSP was averaged at 60 second intervals and expressed as a percentage of the average fEPSP slope measured during the 30 minute reference time, which was expressed as 100%. The response of fEPSP to the same week-old control group was also measured using the same method.

Example 2. When sildenafil and a ROCK inhibitor were co-administered, confirmation of improvement in cognitive function (long-term memory) decline through the Barnes maze test

Using MCAo30, an animal model of acute stroke (cerebral infarction), i) sildenafil or ROCK inhibitor alone, ii) sildenafil and ROCK inhibitor combined administration, to determine the degree of cognitive decline, one of the sequelae due to stroke. , Barnes maze test was performed, and cognitive function was based on the escape time of the normal control group. The experiment was performed based on the method described in 1-4.

As a result, as shown in FIG. 3, it was confirmed that the escape time to find the escape hole of the stroke (cerebral infarction) animal model (MCAo30) increased by about 3 times compared to the normal control group. The above results confirm that memory loss is one of the most serious sequelae following stroke.

In addition, the escape time of the sildenafil alone administration group and the ROCK inhibitor alone administration group was measured at a level similar to that of the normal control group, and it was confirmed that cognitive decline was significantly improved when sildenafil or ROCK inhibitor alone was administered.

In addition, the escape time of the group administered with sildenafil and ROCK inhibitor was measured at a level similar to that of the normal control group, confirming that cognitive decline was significantly improved. It was confirmed that the cognitive function (improvement of memory) was improved compared to that of the normal control group.

In addition, the escape time of the drug-administered group after reperfusion was significantly shorter than that of the drug-administered group after middle cerebral artery occlusion and reperfusion, and it was confirmed that the effect of improving cognitive function was excellent in the drug-administered group after reperfusion after middle cerebral artery occlusion. .

Based on the above experimental results, it can be seen that co-administration of sildenafil and a ROCK inhibitor after reperfusion after middle cerebral artery occlusion can not only improve cognitive decline due to stroke more effectively, but also enhance cognitive function compared to the normal control group. have.

Example 3. When sildenafil and a ROCK inhibitor were administered in combination, it was confirmed whether neuroplasticity was improved

Using MCAo30, an animal model of acute stroke (cerebral infarction), an experiment to confirm the improvement of neuroplasticity when i) sildenafil was administered alone before reperfusion after middle cerebral artery occlusion, ii) sildenafil was administered alone after reperfusion after middle cerebral artery occlusion Was performed, and the cognitive function was compared with the normal control group. The experiment was performed based on the method described in 1-5.

As a result, as shown in FIGS. 4 and 5, it was confirmed that the stroke model MCAo30 deteriorated the fEPSP slope to 84.11%.

However, it was confirmed that the fEPSP slope improved to 102.35% in the drug-administered group after middle cerebral artery occlusion and before reperfusion, almost the same level as the normal control group, and in the drug-administered group after middle cerebral artery occlusion, the fEPSP slope improved to 158.31%. .

Comparing the experimental results numerically, the fEPSP slope was improved by 4.06 times in the drug-administered group after reperfusion after middle cerebral artery occlusion and reperfusion after middle cerebral artery occlusion, and neuroplasticity in the drug-administered group after reperfusion after middle cerebral artery occlusion. It was confirmed that the improvement effect of was significantly improved, and the result was consistent with the result of Example 2.

In addition, when the above experimental results are summarized, it can be seen that the improvement effect of reducing neuroplasticity due to stroke is maximized when sildenafil is administered alone after reperfusion after middle cerebral artery occlusion in an animal model of stroke (cerebral infarction).

The above-described description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains can understand that it is possible to easily transform it into other specific forms without changing the technical spirit or essential features of the present invention. There will be. Therefore, it should be understood that the embodiments described above are illustrative and non-limiting in all respects.

Claims (5)

Phosphodiesterase type 5 activity inhibitor (phosphodiesterase type 5 inhibitor); And
A pharmaceutical composition for the treatment of cognitive decline due to ischemic cerebrovascular disease, comprising a ROCK inhibitor (Rho-associasted kinase inhibitor).
The method of claim 1,
The ischemic cerebrovascular disease is any one selected from the group consisting of ischemic stroke (cerebral infarction), thrombosis, embolism, transient ischemic attack, leukemia, and small infarction, a pharmaceutical composition for the treatment of ischemic cerebrovascular disease.
The method of claim 1,
The phosphodiesterase type 5 activity inhibitors are mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, and dasantafil. , Avanafil (avanafil), and a pharmaceutical composition for the treatment of ischemic cerebrovascular disease, characterized in that at least one selected from the group consisting of a pharmaceutically acceptable salt, solvate, and hydrate thereof.
The method of claim 1,
The ROCK inhibitor is any selected from the group consisting of Fasudil, Ripasudil, RKI-1447, Y-27632, GSK429286A, Y-30141 and pharmaceutically acceptable salts, solvates, and hydrates thereof. A pharmaceutical composition for the treatment of ischemic cerebrovascular disease, characterized in that at least one.
The method according to any one of claims 1 to 4,
The pharmaceutical composition is for administration after cerebrovascular occlusion before reperfusion, simultaneously with reperfusion, or after reperfusion.

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