Pharmaceutical composition for treating ischemic insult and method for therapy

FIELD: pharmaceutical chemistry, medicine.

SUBSTANCE: in the suggested composition one should apply heptapeptide of Met-Glu-His-Phe-Pro-Gly-Pro sequence (heptapeptide A) for treating ischemic insult due to introducing 2 drops of compositions into each nasal canal 5-6 times daily for 10 d at disease of average severity degree, and in case of severe degree - per 3 drops of the present composition into each nasal canal 7 times daily for 10 d. The present innovation provides increased efficiency at decreased concentration of heptapeptide without any side effects.

EFFECT: higher efficiency of therapy.

2 cl, 6 dwg, 8 ex, 5 tbl

 

The invention relates to medicine, namely to the study and application of peptide sequence Met-Glu-His-Phe-Pro-Gly-Pro (heptapeptide a) neuroprotective activity and the creation of a pharmaceutical composition with a neuroprotective effect. Drug on this basis can be used in various neurological diseases associated with brain damage, vascular lesions of the brain, progressive disease of the Central nervous system including hereditary.

The urgency of development of various neuroprotective funds, protection of the Central nervous system through activation of various metabolic processes of the brain, determined by the needs of modern health care. Enough, for example, to say that the mortality from stroke in our country came in second place after cardiovascular diseases. One promising neuroprotective therapy in ischemic tissues is increased neurotrophic support brain, which hinders the development of both apoptotic and necrotic changes of neurons.

Modern understanding of the plasticity of the Central nervous system suggests a significant opportunity to restore its functions, except in the pathological damage to the s, such as trauma, stroke, ischemia, etc. it Is believed that under these influences the cells of the nervous system undergo death by mechanisms of necrosis and programmed cell death (apoptosis). Around the necrotic lesion develops in the region, the cells which are in a state of apoptosis. A distinctive feature of apoptosis is its reversibility, which means that restore the health of nerve cells under the action of a so-called neuroprotective factors. Currently isolated and studied a number of such factors that protect neurons from the damaging effects, in particular, nerve growth Factor (NGF) and Neurotrophic factor brain (BDNF) and others. Moreover, it shows an increase in the level of synthesis of the neurotrophic Factor in the brain following ischemic brain damage. In this regard, there is a possibility of preventing the consequences of pathological effects on the Central nervous system with the help of such neuroprotective factors. There is, however, the principal difficulty of their use in clinical practice because of the way the introduction of these substances into the brain, due to impermeability to them the blood-brain barrier. In this regard, it is important synthesis of short peptides that were able to stimulate the synthesis of barotropically factors in the brain in vivo, especially after intranasal such peptide compounds.

Known heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro as a stimulant memory prolonged action (Patent USSR No. 939440, C 07 C 103/52, And 61 To 37/02, the priority of January 6, 1981) [1], which is the basis of nootropic drugs and the pharmaceutical composition nootropic action (Patent RF № 2045958, 16 a 61 K 38/08, 1994) [2].

There is a method of treatment of stroke using heptapeptide And for patients with moderate and severe strokes in a daily dose, respectively 9-13 and 15-24 mg (Patent RF № 2124365, C1 a 61 K 38/08) [3]. Heptapeptide And in the case of treatment of stroke, moderate and severe strokes used in the form of 4-5% solution (Patent RF № 2124365, C1 a 61 K 38/08). This patent is a prototype.

However, the use of 4-5% solution under the above doses involves introduction 1 drop in each nostril 2 times a day in case of stroke moderate and 1 drop 4 times (or 2 drops 2 times per day in case of a severe stroke. The introduction of the bow one drop of the drug dramatically reduces the efficiency of its use, as only a small part of medication moisten the mucous membrane of the nose. This is the main drawback of the prototype, as in the case of treatment of neurological diseases the drug is advisable to introduce 2-3 drops in each nostril DL is better wetting of the mucous membrane of the nasal cavity in 2-4 hours. In addition, the proposed prototype structure does not determine the pharmacological composition of the medicinal product, which depends on the guaranteed term safety of the drug.

The technical result achieved in the implementation of this invention, is the ability to create a new drug for the treatment of various pathologies of the brain (stroke, ischemia, injury, etc).

This is achieved by the fact that known heptapeptide, the General formula Met-Glu-His-Phe-Pro-Gly-Pro is used as neuroprotective agents in the following pharmaceutical compositions are:

Heptapeptide And distilled water as preservatives nipagin in the following ratio of components, g/l:

heptapeptide mentioned formula of 9.5 to 10.6

nipagin 0,90-1,10

distilled water up to 1 l

The following examples confirm the neuroprotective properties of heptapeptide And neuroprotective properties of drugs and pharmaceutical composition.

Example 1:

Conducted to study the ability of the above peptide sequence Met-Glu-His-Phe-Pro-Gly-Pro (heptapeptide A) to increase the survival rate of cultured nerve cells after hibernation embryonic brain. Nonlinear white rats with pregnancy 16-18 days was scored by carbon dioxide asphyxia. From the brain of embryos were allocated asalnya kernel forebrain, which dissociatively into individual cells. The cell suspension was transferred into a culture medium MEM/F-12 (1:1), products containing 6 g/l D-glucose, 25 mg/l insulin, 100 mg/l transferrin, 20 nm progesterone, 100 nm of putrescine, and 30 nm sodium Selenite, and were sown in 96-well plates, pre-treated D-polylysine. Cell density was 30-50 thousand cells per well. After attachment of cells to the substrate was added solutions heptapeptide and nerve growth Factor (positive control) in the culture medium of the composition to a final concentration of 10 μm and 100 ng/ml, respectively. As control was used environment that does not contain heptapeptide and nerve growth Factor. Cells were cultured in the CO2-incubator (5% CO2) at 37° With, then estimated the number of living cells using staining MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide). Heptapeptide and nerve growth Factor had no effect on cell survival after 24 hours after the start of cultivation, with an immediate allocation of the basal forebrain nuclei from embryonic brain. However, significantly increased the survival rate at pre-incubation dedicated the whole brain in Hanks solution at 4-6° within 24 hours (figure 1). Thus, heptapeptide And slows the death of a part of cultured cells basal the output nuclei of the forebrain, caused by hibernate embryonic brain and exerts a neuroprotective effect comparable to the effect of nerve growth Factor.

Example 2:

Conducted to study the ability of heptapeptide And to increase the survival rate of cultured over a long period of time, the nerve cells. Cultures were obtained and cultured as described in example 1. After attachment of cells to the substrate solution was added heptapeptide And in the culture medium of the composition to a final concentration of 10 μm. As control was used environment that does not contain heptapeptide A. within 7 days after the start of cultivation, the cells were fixed with 4% solution of paraformaldehyde in phosphate-buffered saline and spent the visualization of cholinergic neurons using cytochemical staining of cells on the acetylcholinesterase according to the methodology described in (Hefti F., Hartikka, J., et. Al.Sanchez-Ramos J.//A Dissection and Tissue Culture Manual of the Nervous System./eds.: Shahar, A., de Vellis J./Wiley-Liss, New York. 1989. P.172-182.) [4]. The results of these experiments are shown in figure 2. Heptapeptide And significantly increased the number kholinergicheskikh neurons.

Example 3:

Conducted to study the ability of heptapeptide And influence on the proliferation of glial cells. The culture of glial cells of the basal forebrain nuclei of the rat were obtained from newborn rats according to standard methods. Cells Kul who was rivervale in MEM medium with 15% fetal serum (ES) in CO 2-incubator (5% CO2) at 37° C. the Inclusion of [3H]thymidine into the DNA of glial cells was determined according to the standard technique. Glial cells were sown on 24-hole tablets with a density of 70 thousand cells per well, kept for 1 day in serum-free medium MEM in CO2-incubator (5% CO2) at 37° With, then add these 2 compounds and [3H]thymidine (mccoury/ml) or [3H]thymidine (control). Incubation with added compounds and [3H]thymidine was carried out for 24 hours under similar conditions. Was no significant decrease or increase the inclusion of [3H]thymidine into DNA glial cells (figure 3), indicating the absence of heptapeptide And ability to suppress or stimulate the proliferation of glial cells.

Example 4:

Examined the ability of the peptide sequence Met-Glu-His-Phe-Pro-Gly-Pro (heptapeptide A) to stimulate the synthesis of neurotrophic factor isolated from brain (BDNF) in the hippocampus of the rat brain in vivo. In the work were used male Wistar rats, weighing 300-350 g Animals were kept in standard vivarium conditions with free access to food and water and a 12-hour cycle of light. The peptide was injected animals intranasally in a volume of 100 ál/kg animal body weight once. Control animals were injected with equivalent about the eating of distilled water. The dosage used correspond to those that were effective in behavioral tests. In each group there were 3 animals. To determine the level of BDNF expression in the brain regions of heptapeptide were administered intranasally at doses of 50 and 250 mg/kg of the Experimental animals was deceptional after 3 and 24 hours after injection of the peptide. The relevant parts of the brain (the hippocampus) was separated and frozen in liquid nitrogen. Preparation of extracts from selected departments and further quantitative determination of the level of BDNF using enzyme immunoassay was performed according to the Protocol of the company Promega USA. For each experimental point used 4 parallel. The validity of the results was assessed by student's criterion using the program Sigma Plot 2.01.

From the results shown in figure 4, it is seen that heptapeptide increases levels of BDNF in the hippocampus of the brain of experimental animals. It should be noted that the level of BDNF was increased after 3 hours after injection of heptapeptide. The effect of heptapeptide were detected in the hippocampus and in the 24 hours after its introduction. Increased relative to the control level of BDNF was observed as with the introduction of heptapeptide dose of 50 and 250 mg/kg of body weight of the animal.

Example 5:

Studied the effectiveness of the peptide sequence Met-Glu-His-Phe-Pro-Gly-Pro (heptapeptide A) in vivo in models of global Isha the AI brain in animals with irreversible bilateral occlusion of common carotid arteries. Under the influence of heptapeptide And significantly reduces the severity of neurological deficit and there was a trend of increasing survival of animals, which is associated with the neuroprotective effect of the drug (figure 5).

A randomized blind study was conducted on 40 male Wistar rats, weighing from 80 to 110, Irreversible bilateral occlusion of common carotid arteries was performed under ether anesthesia using a surgical access and simultaneous ligation of arteries silk ligature. Surgery each animal was carried out as a standard, and took no more than 7-10 minutes, then rats recovered quickly after ether anesthesia.

All investigated substances, below, was administered as an intraperitoneal injection (daily dose in 1 ml solution). In the experiment included a group of animals treated with heptapeptide sequence Met-Glu-His-Phe-Pro-Gly-Pro (heptapeptide A), Cerebrolysin, saline (control), as well as linopirdine animals. The study group consisted of 10 animals treated with heptapeptide And the rate of 300 mg/kg/day, fractionally after 15, 60, 120 minutes and 5 hours after occlusion. Dose and fractional appointment of the drug were selected on the basis of clinical and experimental data have demonstrated the neuroprotective effect of small doses of heptapeptide And (3-30 mgkg) and antihypoxic and neurotrophic effects of high doses (150-300 mg/kg) [5,6,7]. The comparison group consisted of 10 animals treated with Cerebrolysin. Active active principle of the drug are low molecular weight peptides with distinct neurotrophic effect [8]. Cerebrolysin was administered at a dose of 2.5 ml/kg/day after 15 and 60 minutes after occlusion. 10 animals in the control group after occlusion received saline at 0.5 ml after 15 and 60 min after ischemia. In the group lonaprisan animals under the same anesthesia was carried out surgical access to the common carotid arteries without ligation, and were injected intraperitoneally with saline. Assessment of neurological deficit was performed blindly (without information about the distribution of rats in groups) every 30 minutes during the day on scales scoring: the definition of Stroke-Index no C.P.McGrow [9] and neurological scale Rudolphi et al. [10]. On a scale C.P.McGrow [9] Stroke-Index increases with the appearance of some signs of neurological deficit, on a scale Rudolphi et al. [10] neurological score varies from 9 (normal) to 0 (death). The main signs of neurological deficits included limiting the mobility of the animal, ptosis (bilateral or unilateral), circling, hyperactive behavior, rotational convulsions, tonic and clonic convulsions, coma with a weak pain response or lack of it.

For statistical analysis Yes is data on the dynamics of neurological deficit was used dispersion factor analysis ANOVA. To assess mortality and separate neurological manifestations were used the Fisher test.

In the control group of animals in the first 3 hours after occlusion of the common carotid arteries rotational and clonic convulsions have been reported in 80% of animals. In groups of animals treated with heptapeptide and Cerebrolysin at 40% (p=0,08) and 50% (p=0,17), respectively.

When assessing neurological deficit during the day revealed that the introduction of Cerebrolysin had no significant influence on the dynamics of change in scores on scales assessing Mc Grow [9] and Rudolphi et al. [10] when compared with the control group (figure 5). In the group of animals treated with heptapeptide And, during the day remained lower values of the average score on a scale Mc Grow [8] and higher values for Rudolphi et al. [7] compared with control animals and animals treated with Cerebrolysin. Significant differences were found in the period from 1.5 to 6.5 hours (p=0.03, ANOVA) between the control group and the group of animals treated with heptapeptide A.

When evaluating posutochno mortality should be noted that 50% of fatal cases in the control group and in the group of animals treated with Cerebrolysin was observed during the first 7 hours after the occlusion, whereas in the group of animals treated with heptapeptide And up to 7 hours, there were no reported fatalities. 50% of itelnych cases in the group of animals receiving heptapeptide And was observed in the interval from 8 to 10 hours after ischemia. When assessing the lethality to the end of the first day after global ischemia of the brain received the following indicators: 100% of the animals in the control group, 90% in the group of animals treated with Cerebrolysin, and 70% in the group of animals treated with heptapeptide And (p>0,05). Life expectancy in the control group was 9.1±1.7 hours, in the group of animals treated with Cerebrolysin, - 11,5±2.3 hours and in the group of animals treated with heptapeptide And, up 14.2±2.5 hours. The average score at the time of death on a scale S.R. McGrow [7] in all groups were comparable: 19,4±0.9 in the control group, 19,9±0.5 in group of animals treated with Cerebrolysin, and 20±1.1 in group of animals treated with heptapeptide A. it Should be noted that by the end of the day in the group of animals treated with heptapeptide And, the average score on a scale Mc Grow [7] was 10 points higher than in the group of animals treated with Cerebrolysin, testified to the survival in the background heptapeptide And even animals with severe neurological deficits.

Experimental studies have shown that the use of heptapeptide And global cerebral ischemia animal has a certain positive effect, reducing the severity of neurological deficit during the first 6.5 hours, p and compared with a control group and a group of animals treated with Cerebrolysin. The observed trend of increasing survival of animals, lengthening the period before the first death may also be due to the neuroprotective properties of heptapeptide A.

Example 6:

The shelf life of the drug “Semax - 1.0% solution” was determined by the method of accelerated aging. Solution dosage forms of the drug, which is an aqueous solution of heptapeptide a (1.0%) with addition of 0.1% of a preservative nipagina was under sterile conditions, filled into vials with a plastic tube by pipette 3 ml in a bottle. Made 3 series of drug 75 bottles in each.

Was implemented input control of the drug according to the following criteria: description of the solution, its color, transparency, pH, sterility, determining the quantitative content of the basic substance and nipagina. Quantitative content was performed by high performance liquid chromatography.

2 μl of the drug “Semax-1.0% solution” analyzed for any liquid chromatograph, high pressure, equipped with a UV spectrophotometric detector at 210-230 nm and thermostat columns.

Used for the analysis of the analytical column (e.g., size 4· 150 mm)filled by reversed-phase (C8, 16 or C18) sorbent (type Nucleosil C8, diasorb ST, diasorb C16T, Bond ODS and others), with grain size is 5-7 microns. To increase the lifetime of the column used predalone, filled with the same sorbent. The mobile phase eluent A. the Eluent And was prepared as follows: To 670 ml of buffer And was added 330 ml of methanol for liquid chromatography. Eluent was filtered through a filter with pore size 0.45 µm. Before use, the eluent was degirolami filtration under vacuum, transmission, helium or other suitable method. Shelf life eluent - 1 month at room temperature in a tightly closed container. The feed rate of eluent 800 µl/min Chromatography was performed under the following conditions: the Volume of injected sample to 2 ml.

Detection at a wavelength of 210 nm. The column temperature 35°C. Scope of registration - 2 eop

To determine heptapeptide and nipagina in solution was performed three-fold analysis of the corresponding solution. Approximate retention time of heptapeptide and nipagina in these conditions analysis was 9.1 min and 18 min, respectively. The desired concentration of heptapeptide and nipagina in solution (Ci,heptand Ci nip) is determined by the formula:

Ci,hept=Ai,hept·Tohept/Vi[mg/ml],

Ci nip=Ai nip·Tonip/Vi[mg/ml],

where ai, heptand ai nip- the peak areas of heptapeptide and nipagina on the i-th chromatogram;

Toheptand K nip- average values of the calibration coefficient for heptapeptide

And nipagina obtained by calibration.

Vi- volume of added sample µl (Vi= 5 ál).

Average concentrations of heptapeptide and nipagina in the product

“Semax-1.0% solution”

(CR,heptand CWed,nip):

Ci,hept=Σ and (Ci,hept)/3; Ci nip=Σ and (Ci nip_/3 [mg/ml].

Content nipagina in the product should be at least about 0.90 and no more than 1.10 g/l Content heptapeptide And the drug should not be less than 9.5 and not more than 10.6 g/L. Values of the standard deviation for heptapeptide and nipagina in a series of three tests should not be more than 0.02 (2%).

Sample preparation “Semax - 1.0% solution” (3 series 75 flasks) were subjected to accelerated aging, which was achieved by keeping them in a thermostat at a constant temperature of 60°C for a certain period of time.

Method of accelerated aging to determine the shelf life of the drug is based on the fact that the safety of substances in accelerated aging (storage in thermostat at T=60°). These periods was 5; 11; 23; 34 and 46 days, which corresponds to 1 month; 6 months; 1 year, 1.5 years, 2 years of storage in the refrigerator at T=8-10° C. At the expiration of each period of accelerated aging was conducted a series of studies on the same criteria, the input control. The quantitative content of heptapeptide and nipagina was determined by HPLC under the same chromatographic conditions as when the input control of the drug.

Used the method of absolute calibration standard (in this case the method of calculation of the amount of substance in the calibration table corresponding to the chromatogram of the sample of the drug, which has not been subjected to accelerated aging). On the basis of the results obtained in the quantitative determination of heptapeptide and nipagina deadlines accelerated aging taking into account the qualitative assessment of the safety of the drug in all the required criteria, a shelf life for the drug product “Semax - 1.0% solution” - 2 years.

Experimental results the safety of the drug Semax - 1.0% solution” method of accelerated aging are shown in table 1.

Example 7:

In this example, the generalized results of clinical use of the drug Semax - 1.0% solution in a clinic.

We examined 80 patients (38 men, 42 women, mean age of 68.3±1,98 years) with acute ischemic hemispheric stroke. All patients were hospitalized during the first 12 hours after stroke. In 42 patients were diagnosed with astronauta cerebral circulation in the system of the left internal carotid artery, 38 in the system right internal carotid artery.

The main etiological factor in the development of stroke in 56 (70 %) patients was the combination of atherosclerosis in hypertension. 14 (17,5%) stroke patients developed on the background of arterial hypertension, at an altitude of hypertensive crisis. The combination of diabetes, atherosclerosis and hypertension was observed in 2 cases(2,5%), 72 (90%) patients had evidence of coronary heart disease, 16 in the past suffered a myocardial infarction. In 1 patient an acute violation of cerebral circulation developed on the background of acute myocardial infarction (cardiocerebral syndrome), 22 (27.5 per cent) of patients had a permanent form of non-rheumatic atrial fibrillation nature, in 4 patients the stroke developed on the background of paroxysmal atrial fibrillation (5%).

All patients with a clinical picture consistent with localization center of brain lesions of varying severity. At the time of admission status in 44 patients (55%) was moderate, as the clinical picture was dominated focal neurologic disorders of moderate severity. In 36 patients (45%) condition was assessed as severe (table 2). These patients had disorders of consciousness and other cerebral symptoms, autonomic and trophic disorders, and signs of severe neurologic defect. Assessment of pain who's focused on three mutually enriching for informative clinical scales scoring neurological deficit (scale - original (Eyisi, Whiskerton, 1992), Orgogozo (1986), Scandinavian (1985)) at the time of admission of patients, 3rd, 5th and 21st day of the stroke. This helped to unify criteria for the severity of the condition, the severity of cerebral symptoms and neurological deficits for comparative analysis, statistical and correlation processing of the received material. A comparative analysis of the effectiveness of therapy was carried out on 5th (at the end of the acute period) and 21st (at the end of the acute period) the day of the stroke.

The control group consisted of 40 patients, who in the acute period of stroke was conducted comprehensive, standardized therapy aimed at the correction of disorders of the Central and cerebral hemodynamics, hemorheological indices, combating swelling of the brain. Neuroprotective drugs were not used. 40 patients from the first hours of the disease on the background of a comprehensive basic therapy was introduced “Semax-1.0% solution” by 2 drops each nostril 5-6 times a day in case of stroke moderate and 3 drops in each nasal passage 7 times a day in severe stroke. This allowed us to assess the effectiveness of the inclusion of a medicinal product “Semax-1.0% solution in the treatment of acute hemispheric ischemic stroke, as well as to clarify the metabolic aspects of the pathogenesis of ischemic Insa is TA and mechanism of action of a drug. The drug “Semax-1.0% solution” was introduced endonasal within the first 10 days of illness.

On the etiological factors, the characteristics of the onset of disease, duration of disease, age and sex composition of all the analyzed groups were comparable.

When comparing clinical dynamics in the group with use of the drug “Semax-1.0% solution” and in the control were found significant improvement in reduction processes on the background of drug treatment “Semax-1.0% solution”. In patients with moderate (p<0,01-0,05) and severe stroke (p<0,001-0,05). The data are shown in table 3.

In 87.5 % of patients receiving the drug “Semax-1.0% solution”, for stroke was egredientem, with a stable regression of neurological disorders, in any case, there was no progression of the stroke. By the end of the acute period of stroke in 84% of patients seen good recovery of impaired functions with regression of musculoskeletal disorders to a minimum monoparesis, preserving minimal elements of motor aphasia and full recourse disorders of sensitivity.

In severe stroke application in the first hours and days of preparation “Semax-1.0% solution is allowed to reliably accelerate the regression of not only local, but also cerebral and autonomic symptoms. All patients admitted in the state of stun, by the end of 2 with the current came back clear consciousness, the consciousness of the 4 patients who were in the spoor, to the 2nd day came on stage stun, only in one case, the spoor was maintained until the end of the 3 days of the disease. To 5-day stroke in 78% of patients with disorders of consciousness and meningeal symptoms regressed completely. Quantitative assessment of the dynamics of the mean values of cumulative clinical score showed a statistically significant ahead of the pace of recovery against the background of the drug “Semax-1.0% solution” for all three used scales, with differences from control were more pronounced than in the group of patients with moderate disease. As in the group of strokes moderate the most pronounced and significant (p<0,05) was accelerated regression of musculoskeletal disorders. In the group treated with the drug “Semax-1.0% solution” severe patients in 78% of cases were determined regimiento the course of the disease (control group - 32,0%), the remaining patients during the acute period of stroke was remittiruuschem. In the group with use of the drug “Semax-1.0% solution” mortality in severe stroke was 8.7%, while in the control group of 22.3% (p=0.07 by the Fisher test). By the end of the acute period of 5% of the surveyed were recorded good recovery of impaired functions (control - 1,3%), 64% - moderate limitation of functions (the control - 17.4%), severe focal defect persisted in 31% of patients (control 81,3%).

Comparing the effectiveness of different doses of the drug “Semax-1.0% solution” (6) found in the group of patients with moderate stroke greater effectiveness of a daily dose of 12 mg (average 150 μg/kg) compared with lower doses. At the same time in a group of seriously ill greater severity of positive clinical dynamics of the disease occurred in the treatment daily dose of 20 mg (270 μg/kg).

Example 8:

In this example, the generalized results of clinical use of the drug Semax-1.0% solution in another clinic. For study were selected 80 patients (44 men, 36 women, mean age 69,8±2,04 years) with acute ischemic hemispheric stroke admitted to hospital within the first 12 hours after stroke (table 4). In 46 patients were diagnosed with acute violation of cerebral circulation in the system of the left internal carotid artery, 34 - in the system of the right internal carotid artery. The main etiological factor in the development of stroke in 64 (80%) patients was the combination of atherosclerosis, arterial hypertension, the combination of diabetes, atherosclerosis and hypertension was observed in 4 cases (5%), in 8 patients the stroke had cardioembolic character, patients suffered p is the permanent form of atrial fibrillation (10%), in 4 patients the stroke developed on the background of paroxysmal atrial fibrillation (5%). All patients with a clinical picture consistent with cortical-subcortical localization of lesion of brain lesions of varying severity. At the time of admission status in 48 patients (60%) was moderate, the clinical picture was dominated focal neurologic disorders of moderate severity, in 32 patients (40%) condition was assessed as severe, these patients had disorders of consciousness and other cerebral symptoms, autonomic and trophic disorders, and signs of severe neurologic defect. Assessment of patients was carried out in three mutually enriching for informative clinical scales scoring neurological deficit (scale - original (Eyisi, Whiskerton, 1992), Orgogozo (1986), Scandinavian (1985) at the time of admission of patients, 3rd, 5th and 21st day of the stroke. This helped to unify criteria for the severity of the condition, the severity of cerebral symptoms and neurological deficits for comparative analysis, statistical and correlation processing of the received material. A comparative analysis of the effectiveness of therapy was carried out on 5th (at the end of the acute period) and 21st (at the end of the acute period) the day of the stroke.

The control group consisted of 40 patients to the m in acute stroke was a complex, standardized therapy aimed at the correction of disorders of the Central and cerebral hemodynamics, hemorheological indices, struggle with swelling of the brain, without the inclusion of neuroprotective drugs. 40 patients from the first hours of the disease on the background of a comprehensive basic therapy was administered the drug “Semax-1.0% solution in doses and according to the scheme of the previous example. This allowed us to assess the effectiveness of the inclusion of the drug “Semax-1.0% solution in the treatment of acute hemispheric ischemic stroke, as well as to clarify the metabolic aspects of the pathogenesis of ischemic stroke and the mechanism of action of the drug. The drug “Semax-1.0% solution” was introduced endonasal within the first 10 days of illness.

On the etiological factors, the characteristics of the onset of disease, duration of disease, age and sex composition of all the analyzed groups were comparable.

Comparison of clinical dynamics in the group with use of the drug “Semax-1.0% solution” and the control has detected a statistically significant improvement of recovery processes on the background of drug treatment “Semax-1.0% solution in patients with moderate (p<0,01-0,05) and severe stroke (p<0,001-0,05) (table 5).

In ischemic stroke moderate dominated the clinical picture of focal neurological symptoms effective is the drug industry “Semax-1.0% solution” was evident from the first day of the disease and was expressed in a General intensification of the sick, a statistically significant acceleration of the regression of focal symptoms to 5-m and 21-day diseases. In 88% of patients receiving the drug “Semax-1.0% solution”, for stroke was egredientem with a stable regression of neurological disorders that were significantly higher than those in the control group (55.2 per cent). By the end of the acute period of stroke in 80% of patients stated a good recovery of impaired functions (to a minimum monoparesis or reflex asymmetry), and in 36% of patients regression of focal symptoms was full (small stroke). In severe stroke application in the first hours and days of preparation “Semax-1.0% solution is allowed to reliably accelerate the regression of not only local, but also cerebral and autonomic symptoms. All patients admitted in the state of stun, by the end of 2 days came back clear consciousness. To 5-day stroke in 80% of patients with disorders of consciousness and meningeal symptoms regressed completely. In 82% of patients treated with the drug “Semax-1.0% solution”was determined regimiento the course of the disease (control group - 58%). In the group with use of the drug “Semax-1.0% solution” mortality in severe stroke was 10%, whereas in the control group of 28.9% (p=0.07 by the Fisher test). By the end of the acute period, 21% of the surveyed registerof is good elk restoration of disturbed functions (in control - at 5,1%), 40% had moderate limitation functions (control - 14,4%), severe focal defect was still present in 39% of patients (control - 57,1%).

Comparing the effectiveness of different doses of the drug “Semax-1.0% solution” found in the group of patients with moderate stroke greater effectiveness of a daily dose of 12 mg (average 150 μg/kg) compared with lower doses. At the same time in a group of seriously ill greater severity of positive clinical dynamics of the disease occurred in the treatment daily dose of 20 mg (270 μg/kg).

The above examples confirm the neuroprotective properties of the peptide sequence Met-Glu-His-Phe-Pro-Gly-Pro) and the efficiency of the proposed pharmaceutical composition.

Literature

1. Ponomarev-Stepnoi M.A., Nezavibatko V.N., Ashmarin I.P., Kamensky A.A., Antonova L.V. (1982). Patent USSR No. 939440, C 07 C 103/52, And 61 To 37/02, the priority of January 6, 1981.

2. La Andreeva, Ipharmacy, ASI, Nfeed, Vinnikopertika, Tveretina (1994). RF patent № 2045958, 16 a 61 K 38/08. The priority of the invention of March 28, 1994

3. Gusev E.I., V.I. Skvortsova, Zhuravleva EJ, Andreeva L.A., Nezavibatko V.N., Dime I.A., meat eaters NF (1999). RF patent № 2124365 C1 a 61 K 38/08. The priority of the invention may 13, 1997

4. Hefti F., Hartikka, J., Sanchez-Ramos J.//A Dissection and Tissue Culture Manual of the Nervous System./eds.: Shahar, A., de Vellis J./Wiley-Liss, New York. 1989. P.172-182.

5. Asmari the I.P., Nezavibatko V.N., Meat eaters NF, Kamensky A.A., Dime I.A., Ponomarev-Stepnoi M.A., Andreeva L.A., Kaplan YA, Koshelev V.B. have been, Racine T.V. (1997). Journal of higher nervous activity. So 47, Vol. 2, S-430.

6. Volkov A.V., Zarzecki J.V., Postnov, A., Balakina G.N., Kamensky A.A., Ants O., Misharin, GV, Gerenko A.N. (1992). The results of the application of regulatory peptides in the intensive care unit after cardiac arrest in the experiment. Terminal States and postresuscitation organism: pathophysiology, clinical features, prevention and treatment. M., Institute of General reanimatology RAMS. Pp.69-76.

7. Kaplan A.Ya., Kochetova A.G., Nezavibathko V.N., Rjasina T.V., Ashmarin I.P. Neuroscience Research Communications. (1996). Vol. 19, No. 2, P.115-123.

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10. Mc Grow P. Arch Neurol. (1977). Vol. 34.- P. 334-336.

Table 2
The distribution of patients by gender, severity of condition and lesion
The group of patientsLocalization of vascular lesionsTotal
 The system of internal carotid artery 
 Left Right 
 menwomenonlymenwomenonly 
A state of moderate severity >36;>25;>50 (55%) points*1016261261844
Heavy state ≤ 36; ≤ 25; ≤ 50 (45%) points*412161282036
Total14 (33,3%)28 (66,6%)42 (100%)24 (63,12%)14 (36,88%)38 (100%)80 (100%)
* points are given by the original. Scandinavian and scale Orgogozo, respectively

Table 3
Distribution of the examined patients of different groups according to severity of condition
The group of patientsA state of moderate severity (>36;>25;>50)Hard state (≤ 36; ≤ 25; ≤ 50)Only
The control group (100%)20 (50%)20 (50%)40
“Semax 1% to dissolve the (100%) 24 (60%)16 (40%)40
Total (100%)44 (55%)36 (45%)80

Table 4
Distribution of the examined patients of different age groups
Age (years)

The group of patients
Younger than 45 years46-5556-6566-75Over 75Only
Control (100%)2(5%)4(10%)14(35%)18(45%)2(5%)40
“Semax-1.0% solution” (100%)2(5%)6(15%)16(40%)14(35%)2(5%)40
Total (100%)4(5%)10(12,5%)30(37,5%)32(40%)4(5%)80

1. Pharmaceutical composition for treatment of ischemic stroke, containing an aqueous solution of heptapeptide formula Met-Glu-His-Phe-Pro-Gly-Pro, characterized in that it contains heptapeptide at a concentration of 0.95-1.06%and additionally nipagin in the following ratio of components, g/l:

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro of 9.5 to 10.6

nipagin 0,90-1,10

the ode distilled else.

2. The method of treatment of ischemic stroke involving the introduction of a pharmaceutical composition according to claim 1, when the disease is moderate to introduce 2 drops of the composition in each nostril 5-6 times a day for 10 days, and the heavy - 3 drops of the composition in each of the bow 7 times a day for 10 days.



 

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