RU2431496C2 - Injection dosage form for treating acute ischemic stroke and craniocereberal injury, method for preparing and using - Google Patents

Abstract

FIELD: medicine, pharmaceutics. ^ SUBSTANCE: invention refers to medicine, and concerns an injection dosage form for treating an acute ischemic stroke and a craniocereberal injury characterised by the fact that as an active ingredient, it contains a therapeutically effective amount of heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro, and at least one substance chosen from a group of antioxidants. ^ EFFECT: invention provides high neuroprotective effect in acute severe CNS affections associating various models of the stroke and the craniocereberal injury, as well as high stability and prolonged use. ^ 12 cl, 13 ex, 2 dwg, 5 tbl

Description

The invention relates to medicine, namely to the treatment of dysfunctions of the central nervous system (CNS) caused by ischemic and traumatic lesions, as well as the field of pharmacology, namely to the development and manufacture of a new injectable dosage form of a previously known substance - heptapeptide of the Met-Glu sequence His-Phe-Pro-Gly-Pro.

The treatment of disorders of the central nervous system caused by ischemic and traumatic brain lesions is a complex task. One of the promising modern approaches is neuroprotective therapy, aimed at improving the survival of neurons in the affected area and normalizing neuroplastic processes in the brain as a whole. Among neuroprotective drugs, various classes of substances and various dosage forms are known. So, for example, a number of drugs with neuroprotective properties (piracetam, noopept, gliatilin) are presented in tablet form. However, the use of tablet neuroprotective agents is ineffective in acute conditions associated with loss of consciousness (stroke, severe traumatic brain injury) and in young children.

The peptide Met-Glu-His-Phe-Pro-Gly-Pro is known as a memory stimulant of prolonged action (USSR Patent 939440, class C07C 103/52, 1981). The possibility of using this heptapeptide as a treatment for ischemic stroke with intranasal administration is described (RF patent No. 2144365, 1997/1999). Intranasal administration of neuroprotective agents, including peptide nature, is a promising direction (Semax preparations - heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro, Selank). This method of administration has a number of positive aspects, such as high bioavailability, low dosage, and painlessness for the patient upon administration. However, one of the limitations of this method of administration is the need for frequent (up to 7 or more times a day) administration of the drug, technical difficulties in administering large doses, which, in general, makes it difficult to use intranasal administration in the treatment of acute severe CNS lesions.

The most widespread in medical practice are injectable peptide neuroprotective drugs, such as cortexin, cerebrolysin, the injectable form of the drug noopept is known (RF patent No. 2330680, 09/12/2005). Injection administration is most consistent with the goals and conditions of emergency therapy for acute severe central nervous system lesions such as stroke and traumatic brain injury (TBI), and the practice of renimation measures. Thus, the use of the heptapeptide of the Met-Glu-His-Phe-Pro-Gly-Pro sequence as part of the Semax preparation for intranasal administration in acute severe central nervous system lesions such as ischemic stroke and head injury has a number of significant differences and disadvantages:

- when using intranasal administration, frequent (up to 7 or more times a day) administration of the drug to the patient is necessary;

- there are technical difficulties with the simultaneous administration of the recommended therapeutic doses by the intranasal method;

- the effectiveness of intranasal administration and the bioavailability of the drug is largely determined by the condition of the nasopharyngeal mucosa of the patient, while the intranasal administration of the existing dosage form is not justified for colds, traumatic or other lesions of such;

- storage of the peptide preparation in a solution for intranasal administration leads to reduced stability of the active substance, which ensures a shelf life of two years and requires a special temperature regime of + 8-10 ° C.

It is known to use the Semax peptide of the formula Met-Glu-His-Phe-Pro-Gly-Pro as an antithrombolytic, anticoagulant, fibrinodepolymerization and fibrinolytic agent for intravenous administration (RF patent No. 2286168). In this case, it was proposed to dissolve 1 mg of Semax - Met-Glu-His-Phe-Pro-Gly-Pro in 1.5 ml of physiological saline and 0.5 ml (200 μg) per 200 g of body weight administered to laboratory rats. However, the use of such a solution of the peptide has several disadvantages associated with the instability of the peptides in the solution in general, and Semax in particular in solutions, in the absence of stabilizing and buffering components. In addition, there is currently no dosage form that allows the implementation of the proposed procedure in the clinic. Therefore, the results presented in the patent of the Russian Federation 2286168 have experimental value, but cannot be applied in practical medicine.

The present invention is the creation of an injectable dosage form of the drug Met-Glu-His-Phe-Pro-Gly-Pro, which is suitable for the treatment of acute ischemic stroke and traumatic brain injury and can be stored for a long time in the form of a solution and emulsion for direct administration, and also in lyophilized form and in the form of dry powder.

Another objective is to increase the effectiveness of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro for the treatment of acute ischemic stroke and traumatic brain injury.

The tasks are solved by the fact that the proposed dosage form based on the heptapeptide sequence Met-Glu-His-Phe-Pro-Gly-Pro for the treatment of acute ischemic stroke and traumatic brain injury, which according to the invention contains a therapeutically effective amount of heptapeptide as an active substance Met-Glu-His-Phe-Pro-Gly-Pro and at least one substance selected from the group of antioxidants including β-carotene, retinol acetate, α-tocopherol acetate, ascorbic acid, lutein, quercetin, rutin (vitamin P), dihydride rockvercetin, butylhydroxytoluene, α-lipoic acid, L-carnitine, succinic acid, as well as optionally at least one excipient selected from the group of stabilizers, prolongators, buffering agents, emulsifiers - solubilizers, solvents, fillers, preservatives and other excipients approved for medical use.

The proposed composition contains mainly the following ratio of components, wt.%:

heptapeptide 0,0002-99,99998, antioxidant 0.00002-99.9998, Excipients up to 100.

Moreover, as a stabilizer, the composition contains at least one substance selected from the group comprising Trilon B, sodium metabisulfite, sodium thiosulfate, glycine, arginine, histidine, lysine or their physiologically acceptable salts, for example, such as hydrochloride, sulfate, acetate, glutamate, aspartate and maleate;

at least one substance selected from the group consisting of polyvinylpyrrolidone with a molecular weight of 10-60 kDa, dextran with a molecular weight of 10-100 kDa, for example, polyglukin or reopoliglukin, polyvinyl alcohol, sodium carboxymethyl cellulose;

as a buffering agent, at least one substance selected from the group consisting of sodium chloride, sodium / potassium hydro and / or dihydrogen phosphate, sodium or ammonium acetate;

as an emulsifier - solubilizer - at least one substance selected from the group comprising soya lecithin for injection, polysorbate-20, polysorbate-60, polysorbate-80, span-20, span-40, span-60, span-85 sodium dodecyl sulfate;

as a solvent, at least one substance selected from the group comprising water for injection, olive oil, peach oil, sunflower oil;

as a filler, at least one substance selected from the group comprising sorbitol, mannitol, xylitol, lactose, sucrose, dextrose, copolymer D, L-lactic and glycolic acids;

at least one substance selected from the group consisting of chlorobutanol hydrate, ethyl alcohol, benzyl alcohol, phenol, chlorocresol, benzoic acid, sorbic acid, merthiolate, nipagin as a preservative; nipazole, benzalkonium chloride, benzethonium chloride, cetylpyridinium chloride, dimethyldodecylbenzylammonium chloride.

The proposed injectable dosage form may be in the form of a lyophilized powder, dry powder, emulsion, solution for intramuscular or intravenous administration, or a solution for infusion.

The essence of the invention lies in the fact that, as it was experimentally established, the addition of at least one of the above antioxidants to the Met-Glu-His-Phe-Pro-Gly-Pro peptide significantly potentiates the pharmacological activity of this peptide, which allows to reduce the concentration of the peptide in form and get a more pronounced pharmacological result.

Thus, the proposed injection form has properties that enhance the effectiveness of therapy (severity of neuroprotective effect) of acute severe CNS lesions, such as ischemic stroke and traumatic brain injury, by increasing the bioavailability of the peptide, reducing the frequency of administration of the drug during the day, and facilitating the administration therapeutic dose and ensuring the accuracy of dosing of the necessary therapeutic dose.

In addition to the above therapeutic advantages, the proposed injection form is stable at room temperature, has a longer shelf life (at least 3 years) compared with the dosage form for intranasal administration, and does not require special temperature conditions for storage and transportation.

The following possible compositions of the injectable peptide form Met-Glu-His-Phe-Pro-Gly-Pro, developed experimentally, are proposed.

In the form of dry powder on one ampoule or bottle:

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.001-0.02 g routine 0.0005-0.025 g, glycine 0.00-0.025 g lactose 0.005-0.50 g; or heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.001-0.020 g, ascorbic acid 0.0005-0.050 g sodium chloride 0.0001-0.01 g, sodium hydrogen phosphate 0.0001-0.005 g, sodium dihydrogen phosphate 0.0001-0.0025 g, mannitol 0.005-0.5 g; or heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.001-0.020 g, ascorbic acid 0.0005-0.050 g α-lipoic acid 0.0005-0.025 g, glycine 0.005-0.050 g.

In the form of a solution for one ampoule or bottle:

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.001-0.020 g, quercetin 0.0005-0.010 g, polyvinylpyrrolidone 0.0005-0.050 g trilon-B 0.00-0.010 g glycine 0.0001-0.050 g water for injection up to 1 ml.

In the form of a solution for infusion on one bottle:

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.001-0.020 g, ascorbic acid 0.0001-0.500 g succinic acid 0.0001-0.300 g lysine 0.00-0.03 g dextran with a molecular weight of 10-100 kDa 0.00-12.0 g 0.9% sodium chloride solution isotonic or glucose solution 5-10% 100.0-500.0 ml.

As bottles or containers for infusion, bottles or containers made of polypropylene can be used.

In the form of an emulsion per ampoule or vial:

Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.001-0.02 g α-tocopherol acetate 0.0005-0.050 g Polysorbate-80 0.0001-0.010 g, Nipazole 0.0005-0.0001 g, Water for injection 0.1-0.5 ml Olive oil 1.5-1.9 ml.

A method for preparing a dosage form for injection by mixing the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro with an antioxidant and at least one suitable excipient, and, if necessary, sterilizing the injectable dosage form; pouring the solution into ampoules, vials or containers for infusion solutions; drying solutions in a sublimation unit; sealing of ampoules or corking of finished product bottles.

A wide range of concentrations of excipients is due to the fact that substances with antioxidant activity contribute to the manifestation of a synergistic effect when included in the composition of the injection drug in a wide range of concentrations. In particular, antioxidant additives, even at the lowest doses, significantly stabilize the heptapeptide molecule of the Met-Glu-His-Phe-Pro-Gly-Pro sequence and potentiate its pharmacological activity. With an increase in the concentration of antioxidant in the composition of the drug, the antioxidant not only potentiates the pharmacological effect of the peptide, but also manifests its therapeutic effect, which further potentiates the pharmacological effect of the drug. The concentration of auxiliary substances of other groups, for example, prolongators (polyvinylpyrrolidone 0.05 - 50.0 mg, dextran from 1.0 to 12.0 grams) is selected in accordance with the method of application and dosages in clinical practice.

The proposed composition of the drug allows it to be used for intramuscular and intravenous injections, as well as in the composition of infusion solutions, which allows the use of new dosage forms in cases where the use of the intranasal form is not effective. Thus, the use of the drug in therapy creates convenience during emergency resuscitation.

The drawings are presented, confirming the receipt of the technical result, in which Fig. 1 shows the dynamics of the neurological status in rats with incomplete global ischemia during experimental therapy with the proposed dosage forms based on Met-Glu-His-Phe-Pro-Gly-Pro (n = 15, * - p≤0.05 in comparison with the control group (saline) and 1% solution of Met-Glu-His-Phe-Pro-Gly-Pro) for example 9;

figure 2 shows the mortality curves of rats in groups with incomplete global ischemia during experimental therapy with new dosage forms based on the substance Met-Glu-His-Phe-Pro-Gly-Pro (n = 15, * - P ₁ = 0.002; P ₂ = 0.010; P ₃ = 0.012; P ₄ = 0.009; P ₅ = 0.05 with respect to the control group, respectively) for example 11.

The following are examples of the preparation of heptapeptide dosage forms of the Met-Glu-His-Phe-Pro-Gly-Pro sequence for injection: liquid forms (solutions and emulsions for injection), solid forms (dry sterile powders, lyophilized powders for the preparation of solutions for injection) The possible compositions of each type of injectable dosage form and a description of the properties of the resulting preparation are given.

Example 1. Obtaining a dry sterile powder for injection.

To obtain a stable injectable heptapeptide dosage form, the Met-Glu-His-Phe-Pro-Gly-Pro heptapeptide sequences are prepared by dry crushing under aseptic conditions using trituration from sterile heptapeptide and excipients. The above sorbitol, mannitol, xylitol, lactose, sucrose, dextrose, copolymer of D, L-lactic and glycolic acids can be used as a filler in the composition for obtaining dry powder; substances that have a stabilizing effect on the heptapeptide molecule (glycine, arginine, histidine, lysine or their physiologically acceptable salts) and other auxiliary substances approved for medical use.

The particular pharmaceutical composition was in sterile solid form, in the form of a dry powder. The drug in appearance is an amorphous powder or a porous mass of white with a yellowish tint or yellow in an ampoule or vial. The ampoule or vial contains

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.0015 g routine 0.0075 g glycine 0.01 g lactose 0.30 g.

When used, the contents of the ampoule or vial are dissolved in 1-2 ml of water for injection or isotonic 0.9% sodium chloride solution or in 0.5-1% procaine solution immediately before injection.

Example 2. Obtaining a dry sterile powder for injection.

To obtain a stable injectable heptapeptide dosage form of the Met-Glu-His-Phe-Pro-Gly-Pro heptapeptide, two powders of the following compositions are prepared separately under aseptic conditions:

A) mixing sterile heptapeptide sequences of Met-Glu-His-Phe-Pro-Gly-Pro and an excipient of the above.

B) mixing an auxiliary substance from the group of antioxidants with buffering additives. As antioxidants in the composition for producing dry powder, ascorbic acid, rutin (vitamin P), lutein, quercetin, dihydroquercetin, α-carotene, butylhydroxytoluene, α-lipoic acid, L-carnitine, succinic acid can be used. Sodium chloride, sodium / potassium hydro- and / or dihydrogen phosphates, sodium or ammonium acetates and other auxiliary substances approved for medical use can be used as a buffering additive in the composition for obtaining dry powder.

The obtained powders A and B are combined and packaged in ampoules or vials under aseptic conditions.

The specific pharmaceutical composition was made in solid sterile form, in the form of a dry powder and in appearance it was an amorphous powder or a porous mass of white or white with a yellowish tint in an ampoule or vial.

The ampoule or vial contains

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.0025 g vitamin C 0.0050 g sodium chloride 0.003 g sodium hydrogen phosphate 0.003 g sodium dihydrogen phosphate 0.0015 g mannitol 0.2 g

When used, the contents of the ampoule or vial are dissolved in 1-2 ml of water for injection or isotonic 0.9% sodium chloride solution or in 0.5-1% procaine solution immediately before injection.

Example 3. Obtaining an injectable aqueous solution.

To prepare a stable injection solution in water for injection, first dissolve auxiliary substances from the groups of antioxidants, prolongators, stabilizers, etc. As antioxidants, ascorbic acid, rutin (vitamin P), quercetin, dihydroquercetin, β- can be used in the composition for injection water. carotene, α-lipoic acid, L-carnitine, succinic acid. Polyvinyl pyrrolidone with a molecular weight of 10-60 kDa, dextran with a molecular weight of 10-100 kDa (polyglyukin or reopoliglukin), polyvinyl alcohol, sodium carboxymethyl cellulose can be used as prolongators in the composition for injectable aqueous solution. Trilon B, sodium metabisulfite, sodium thiosulfate, glycine, arginine, histidine, lysine or their physiologically acceptable salts (hydrochloride, sulfate, acetate, glutamate, aspartate and maleate) can be used as stabilizers in the composition for injectable aqueous solution. Then, the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro is dissolved in the resulting excipient solution. The resulting solution is sterilized by membrane filtration under aseptic conditions, passing through a filter with a pore diameter of 0.22 μm, poured into ampoules or vials in an inert gas atmosphere, ampoules are sealed or the bottles with finished products are sealed and run-in.

A specific pharmaceutical composition is made in liquid form, in appearance it is a clear liquid, packaged in ampoules or vials. One ampoule / vial contains

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.0035 g quercetin 0.0010 g polyvinylpyrrolidone 0.0015 g trilon-B 0.0010 g glycine 0.0050 g water for injections up to 1 ml.

The resulting preparation is ready for direct injection.

Example 4. Obtaining an injection solution for infusion.

To prepare a stable injection solution for infusion in water for injection, auxiliary substances from the groups of prolongators, stabilizers, antioxidants are alternately dissolved. The heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro is dissolved in the resulting excipient solution.

A solution containing all the components is sterilized by membrane filtration under aseptic conditions on a filtration unit, pre-filtering through cartridges with a pore diameter of 0.45 μm and sterilizing filtration through membranes with a pore diameter of 0.22 μm. A sterile injection solution for infusion is poured into bottles or containers made of polypropylene for infusion solutions, cork and run the bottles, seal containers with finished products.

The pharmaceutical composition is made in liquid form, in appearance it is a transparent liquid, in a bottle or container made of polypropylene for infusion solutions. The vial contains

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.020 g vitamin C 0.350 g succinic acid 0.250 g lysine 0.020 g dextran (with a molecular weight of 10-100 kDa) 12.0 g 0.9% isotonic sodium chloride solution 200 ml.

The drug is ready for direct use.

Example 5. Obtaining an injection solution for infusion "ex tempore".

In contrast to the method for preparing an injection solution for infusion described in EXAMPLE 4, a technological technique can be used to obtain a stable injection solution for infusion, which allows preparing a solution for intravenous administration in intensive care and resuscitation departments, as well as in the provision of emergency medical care “ex tempore. " For this, an isotonic solution of sodium chloride for infusion is used as a base solution, to which a basic solution is added containing auxiliary substances from the groups of antioxidants or stabilizers and the heptapeptide of the Met-Glu-His-Phe-Pro-Gly-Pro sequence. Polyglukin (6% isotonic dextran solution with an average molecular weight of about 70 kDa) or Reopoliglukin (10% isotonic dextran solution with a molecular weight of 30-40 kDa) can be used as a base solution in the composition for an injection solution for infusion at least one of the above antioxidants.

To prepare a stable injection solution for infusion, the sterile heptapeptide powder of the Met-Glu-His-Phe-Pro-Gly-Pro sequence is dissolved in a bottle in 1-2 ml of a 10% solution of ascorbic acid for injection. The prepared stock solution is injected with a syringe into the vial of the base solution or a polymer container for infusion solutions, observing aseptic rules. Ready infusion solution is administered intravenously.

The “ex tempore” prepared pharmaceutical composition is in liquid form, in appearance it is a clear liquid in a polypropylene polymer bottle or container for infusion solutions. Contained in vial / container

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.020 g ascorbic acid solution 10% 2 ml Polyglukin solution (6% solution molecular weight dextran 10-100 kDa in isotonic sodium chloride solution) 200 ml.

Example 6. Obtaining an injection solution in the form of an emulsion.

To reduce the hydrolytic processes upon contact of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro with water and to include antioxidants exhibiting hydrophobic activity in the preparation, non-aqueous solvents, in particular fatty ones, can be used as a solvent for the injection oils (olive oil, peach oil, sunflower oil), and substances that make it possible to obtain homogeneous diphilic systems from the group of emulsifiers (solubilizers). At least one substance selected from the group consisting of soya lecithin for injection, polysorbate-20, polysorbate-60, polysorbate-80, span-20, span- can be used as emulsifiers in the composition for injection in the form of an emulsion. 40, span-60, span-85, sodium dodecyl sulfate.

As antioxidants in the composition for injection in the form of an emulsion, any of the above antioxidants can be used. To increase the antimicrobial stability of the drug, one of the above preservatives may be included in the composition. Polysorbate-80, nipazole, an oil solution of vitamin E (α-tocopherol acetate) are dissolved in olive oil. The heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro is dissolved in a minimum amount of water for injection. The resulting aqueous solution of the active substance with stirring is quantitatively transferred to an oil solution containing an antioxidant, preservative and emulsifier.

A specific pharmaceutical composition was made in liquid form, in appearance it is a viscous liquid in an ampoule or vial. The ampoule / vial contains

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.002 g α-tocopherol acetate 0.020 g polysorbate-80 0.010 g nipazole 0.001 g water for injections 0.25 ml olive oil up to 2 ml.

Example 7. Obtaining lyophilized powder for the preparation of a solution for injection.

To obtain a stable during storage and well soluble lyophilized powder for the preparation of a solution for injection, auxiliary excipients from the groups of fillers, stabilizers, buffering additives, and prolongators are used.

The pharmaceutical composition based on the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro can be obtained by any known lyophilization method. One of these methods involves the following steps:

1. Preparation of the filling solution.

- Dissolution of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro in water for injection.

- Addition of antioxidant and excipients.

2. Sterilizing filtration, which includes preliminary and sterilizing filtration on sequentially installed Pall filters with pore diameters of 0.5 and 0.22 microns.

3. Aseptic filling of the solution into ampoules or vials.

4. Freeze-drying of the solution in a freeze-drying unit. The cassettes are placed in a freeze-drying unit and frozen for 4 hours to a temperature of (-45) ° C. Drying is carried out at a residual pressure of from 100 to 120 μm Hg. within 48 hours.

5. Sealing of ampoules, capping and filling of bottles with the finished product.

6. Packaging and labeling.

The pharmaceutical composition is made in sterile solid form, in appearance it is an amorphous powder or a loose tablet in an ampoule or vial.

Each vial or vial contains

heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.002 g vitamin C 0.010 g α-lipoic acid 0.002 g glycine 0.026 g

When using, the contents of the ampoule or vial are dissolved in 1-2 ml of water for injection or isotonic 0.9% sodium chloride solution or in 0.5-1% procaine solution immediately before injection.

Example 8. The effect of injection forms of the proposed composition on the volume of necrosis with the development of focal stroke.

The rats used were Wistar populations. In the experiments, rats were used, aged 14 weeks and weighing 253.2 ± 35.6 g. The quarantine period was 10 days. Rats were kept under standard conditions: a 12-hour adjustable day-night regimen, free access to water and food, 8 rats in a standard T4 cage were mixed — animals of the control and experimental groups. For feeding, we used full-fledged compound feed for keeping rats in vivarium conditions; no additional feeding was introduced. Animals were divided into six experimental groups, 9-11 rats in each group.

The effects of peptide drugs on the volume of brain damage in ischemic stroke were studied using the focal stroke model using a modified technique (Chen, 1964). The operation was performed under anesthesia, 400 mg / kg of chloral hydrate intraperitoneally. Ischemic damage was created by occlusion of the left branch of the middle cerebral artery and a suitable vein with simultaneous ligation of the ipsilateral carotid artery to stabilize the volume of the lesion. The experimental preparations of the peptide Met-Glu-His-Phe-Pro-Gly-Pro were administered intraperitoneally at a dose of 150 μg / kg in a volume of 100 μl per 100 g of animal weight 15 minutes and 2 hours 15 minutes after coagulation of the middle cerebral artery and in the same dose 24 and 48 hours after surgery. A 1% solution of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro was injected in the same dose. The control group was injected with physiological saline in an equivalent volume. After 72 hours, anesthetized (400 mg / kg chloral hydrate intraperitoneally) rats were decapitated, the brain was removed to determine the volume of necrosis. To determine the size of the lesion area of the cerebral cortex, sections 1.5–2 mm thick were stained with 1% solution of 2,3,5-Tripheniltetrazolium chloride (TTX), revealing the dehydrogenase activity of living cells. Brain infarction size was measured planimetrically using the Auc7 program.

Table 1 The size of the lesion of the cerebral cortex in rats with focal stroke Dosage form The size of the lesion (% of the total cortex of the left hemisphere of the brain) one Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 5 wt.%, Ascorbic acid 25 wt.%, Α-lipoic acid 5 wt.%, Glycine 65 wt.%. (example 7) 5.1 ± 0.3 * 2 The heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.47 wt.%, Rutin 2.35 wt.%, Glycine 3.14 wt.%, Lactose 94.04 wt.%. (example 1) 5.3 ± 0.4 * 3 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.35 wt.%, Quercetin 0.10 wt.%, Polyvinylpyrrolidone 0.15 wt.%, Trilon-B 0.10 wt.%, Glycine 0, 50 wt.%, Water for injection up to 1 ml (example 3). 6.4 ± 0.6 * four Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1 wt.%, Α-tocopherol acetate 1.0 wt.%, Polysorbate-80 0.5 wt.%, Nipazole 0.05 wt.%, water for injection 12.5 wt.%, olive oil up to 2 ml (85.85 wt.%) (example 6) 6.0 ± 0.7 * 5 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.0 wt.%, Sodium chloride solution isotonic 0.9% 1 ml. 8.3 ± 1.5 6 Physiological solution (isotonic sodium chloride 0.9% solution) - control 9.5 ± 2.4 Note: * P≤0.05 with respect to groups 5 and 6

According to the results of this series of experiments, it can be concluded that the injectable forms of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro protect the brain of rats from ischemia, the size of necrosis is significantly reduced in comparison with the control groups (6) and the injection of 1% heptapeptide solution Met-Glu-His-Phe-Pro-Gly-Pro in a solution of sodium chloride isotonic 0.9%.

Example 9. A study of the effect of injectable heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro sequences on rat incomplete global brain ischemia models.

The study was conducted on rats of the Wistar population weighing 165.5 ± 15.7 g. Animals were divided into six experimental groups, 15 rats in each group.

Assessment of neurological symptoms and mortality of animals with ischemic damage to rat brain was evaluated in a model of incomplete global brain ischemia (Hossmann K.-A., 1998). Incomplete global brain ischemia was created by irreversible simultaneous ligation of two carotid arteries under ether anesthesia. To prevent reperfusion, two ligatures were placed on each of the two vessels, between which the vessel was cut. Essential anesthesia allows you to almost immediately monitor the development of neurological symptoms and does not cause a protective effect on the brain during the development of ischemia (McGraw, 1977). Peptide Met-Glu-His-Phe-Pro-Gly-Pro peptides were administered intraperitoneally at a dose of 150 μg / kg in a volume of 100 μl per 100 g of animal weight 15 minutes and 2 hours 15 minutes after ligation of the carotid arteries on the day of surgery and in that the same dose once a day for another 4 days after surgery. A 1% solution of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro was injected in the same dose. The control group was injected with physiological saline in an equivalent volume. Within 10 days, weight dynamics and animal mortality were evaluated.

After simultaneous bilateral ligation of the common carotid arteries, a pronounced symptomatology of neurological deficiency develops, which is expressed first in the restriction of the animal’s mobility, the appearance of ptosis (omission of the upper eyelid due to impaired muscle function that lifts it), then violent movements or propulsions: circles and jumps, rotational, clonic and tonic convulsive seizures within a few hours after occlusion of the common carotid arteries, paresis (decrease in the strength or amplitude of voluntary movements due to a violation of the innervation of the corresponding muscles) and paralysis (loss of motor function as a result of pathological processes in the nervous system) of the extremities, then a coma occurs - the animal breathes, but does not respond to irritations, and ultimately the animal dies. The condition of each animal after ligation of the carotid arteries was evaluated on a point scale every 30 minutes for ten hours and the next day. A specific score is assigned to each trait that occurs in animals after ischemia. The accumulated points were summed up every 30 min, the total score characterizes the severity of neurological deficit: 0-3 points - the condition is estimated as close to normal, 3-6 - moderate, 7-24 - severe, 25 points on the Sarkisova scale - death. The result of assessing the development of neurological deficit on the Sarkisova scale is presented in Figure 1. As can be seen from figure 1, the animals of the study groups, which were introduced heptapeptide sequence Met-Glu-His-Phe-Pro-Gly-Pro in dosage forms in accordance with Table 1 to Example 8, have the same average score and the dynamics of the development of neurological status. Statistically, these groups significantly differ from the control group (physiological solution) starting from 2 hours of the development of ischemia and the injection group of 1% Met-Glu-His-Phe-Pro-Gly-Pro solution. Statistical processing of the results suggests that the developed dosage forms based on the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro, administered intraperitoneally at a dose of 150 μg / kg, facilitate neurological status compared with the control group of animals.

Example 10. The frequency of appearance of rats with mild and severe manifestations of neurological symptoms with incomplete global brain ischemia during experimental treatment with injectable dosage forms of the heptapeptide of the Met-Glu-His-Phe-Pro-Gly-Pro sequence.

The experiment was carried out in accordance with the procedure described in Example 9. Relief of the neurological status is due to the appearance of a larger number of animals with a mild form of neurological deficiency in these groups. In this series of experiments, the data processing relied on an estimate of the incidence of rats with mild and severe course of the disease (Table 2). From table 2 it follows that the heptapeptide sequence Met-Glu-His-Phe-Pro-Gly-Pro, introduced in the form of the proposed drugs intraperitoneally in the acute stage of development of ischemic lesion, causes a relief of the neurological status of rats. Injection of a 1% solution of the indicated heptapeptide led to a decrease in severe forms at the trend level.

table 2
The ratio of rats with mild and severe neurological deficit in different experimental groups Experimental groups The number of rats with severe symptoms The number of rats with mild symptoms Statistically significant differences compared with the control group Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 5 wt.%, Ascorbic acid 25 wt.%, Α-lipoic acid 5 wt.%, Glycine 65 wt.% (Example 7) 3 12 P = 0.006 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.17 wt.%, Ascorbic acid 2.34 wt.%, Sodium chloride 1.40 wt.%, Sodium hydrogen phosphate
1.40 wt.%, Sodium dihydrogen phosphate 0.70 wt.%, Mannitol 93.02 wt.% (Example 2) 5 10 P = 0.035 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.35 wt.%, Quercetin 0.10 wt.%, Polyvinylpyrrolidone 0.15 wt.%, Trilon-B 0.10 wt.%, Glycine 0, 50 wt.%, Water for injection up to 1 ml (example 3). four eleven P = 0.035 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1 wt.%, Α-tocopherol acetate 1.0 wt.%, Polysorbate-80 0.5 wt.%, Nipazole 0.05 wt.%, water for injection 12.5 wt.%, olive oil up to 2 ml (85.85 wt.%) (example 6) 6 9 P = 0.009 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.0 wt.%, Sodium chloride solution isotonic 0.9% 1 ml. 7 8 P = 0.052 Physiological solution (sodium chloride solution isotonic 0.9%) - control 13 3

Example 11. Mortality of rats with incomplete global cerebral ischemia during experimental treatment with injectable dosage forms of the heptapeptide of the sequence Met-Glu-His-Phe-Pro-Gly-Pro.

The experiment was carried out in accordance with the procedure described in Example 9. During the acute period of development of ischemia, the injection of heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro heptapeptide sequences prevented rat mortality. In the groups receiving these drugs in the first 12 hours of the experiment, significantly fewer rats died than in the control group (Figure 2). However, the total mortality of animals 10 days after the experiment is the same and does not differ in these groups. Thus, the developed injectable dosage forms significantly expand the “therapeutic window” for ischemic brain damage, but are not a substitute for general resuscitation measures.

Example 12. The effect of experimental therapy with injectable heptapeptide dosage forms of the Met-Glu-His-Phe-Pro-Gly-Pro heptapeptide on the restoration of conditioned reflexes after closed traumatic brain injury (TBI).

The study was conducted on male albino rats weighing 150-160 g. Animals were kept in a vivarium in plastic cages of 5 animals each at an air temperature of 20-22 ° C, natural light, and on bedding from sawdust of deciduous trees. Feeding was carried out ad libitum in the morning. An animal solution of picric acid was used to label animals. The comparability of the experimental groups was ensured by randomizing the samples used in the experiments. The number of animals in the group of 20 pcs.

The simulation of TBI was carried out using the weight-drop method on the original installation. Experimental therapy was started 0.5 hours after mechanical exposure. The experimental preparations were administered intraperitoneally at a dose of 150 μg / kg in a volume of 100 μl per 100 g of animal weight 15 minutes and 2 hours 15 minutes after coagulation of the middle cerebral artery; and at the same dose 24 and 48 hours after surgery. A 1% solution of the heptapeptide sequence of Met-Glu-His-Phe-Pro-Gly-Pro was injected in the same dose. The control group was injected with physiological saline in an equivalent volume.

To study the restoration of mnemonic functions after traumatic brain injury, the method of active swimming avoidance was used. Prior to modeling a closed head injury in experimental animals, a conditioned reflex of active avoidance by swimming with a latent period of 10-20 s was formed for 3 days. In the process of developing reflexes, animals that were not capable of fulfilling the tasks were rejected.

Assessment of the restoration of the pre-formed conditioned reflex was carried out starting from the first day after the application of head injury, and continued daily for 7 days. The time the animals spent the central rod in the pool and the animal left the water was estimated.

The studied drugs with intraperitoneal administration had a positive effect on the recovery time of the conditioned reflex, starting from the 3rd day of the experiment. Injection of a 1% solution of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro also had a positive, but less pronounced effect on the restoration of cognitive functions. The dynamics of recovery of this reflex is presented in the Table. 3.

Table 3
The dynamics of the restoration of the conditioned reflex of active avoidance by swimming (time of animals leaving the water, sec) Experimental groups Study time, days before injury one 2 3 four 5 6 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 5 wt.%, Ascorbic acid 25 wt.%, Α-lipoic acid 5 wt.%, Glycine 65 wt.% (Example 7) 17.4 24.3 9.3 * 6.9 * 5.8 * 6.8 * 5.9 * Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.17 wt.%, Ascorbic acid 2.34 wt.%, Sodium chloride 1.40 wt.%, Sodium hydrogen phosphate 1.40 wt.%, Sodium dihydrogen phosphate 0.70 wt.%, Mannitol 93.02 wt.% (Example 2) 17.4 22.8 8.2 * 5.8 * 6.3 6.2 * 5.1 * Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.35 wt.%, Quercetin 0.10 wt.%, Polyvinylpyrrolidone 0.15 wt.%, Trilon-B 0.10 wt.%, Glycine 0, 50 wt.%, Water for injection up to 1 ml (example 3). 17.4 29.8 8.5 * 7.2 * 7.8 * 7.9 * 6.1 * Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1 wt.%, Α-tocopherol acetate 1.0 wt.%, Polysorbate-80 0.5 wt.%, Nipazole 0.05 wt.%, water for injection 12.5 wt.%, olive oil up to 2 ml (85.85 wt.%) (example 6) 17.4 30.6 9.3 * 6.9 * 7.1 * 6.8 * 5.9 * Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.0 wt.%, Sodium chloride solution isotonic 0.9% 1 ml. 17.4 28.6 13.9 9.5 * 12.1 11.1 * 10.2 * Physiological solution (sodium chloride solution isotonic 0.9%) - control 17.4 31.5 15,5 17.1 12.5 18,4 17,2 Note: * - p <0.05 in comparison with the group "Physiological solution (control)"

Example 12. The study of the synergistic effect of the heptapeptide sequence of Met-Glu-His-Phe-Pro-Gly-Pro and antioxidants in the composition of complex injection drugs on a model of focal brain ischemia in rats induced by photothrombosis.

The work was performed on 80 male Wistar rats weighing 213 ± 2 g. Animals were kept in a vivarium with free access to food and water and a twelve-hour light regime (lighting - from 7:00 to 19:00).

Unilateral focal ischemic infarction in the sensorimotor region of the rat cerebral cortex was created by the method of photochemically induced thrombosis according to the technique described by Watson et al. (Watson et al., 1985). Limb-placing and Forelimbplacing tests were used to determine the neurological deficit caused by photochemically induced thrombosis and assess the effect of drugs on the restoration of impaired functions (motor activity of the limbs, coordination of movements). To visually control the size of ischemic cerebral infarction caused by photothrombosis, the animals were decapitated the day after the last tests mentioned above (on the 8th day after surgery), the brain was removed from the cranium and a visual assessment of the degree of cortical damage in the sensorimotor region was performed.

Limb-placing test. We used a variant of the test, first described by De Rick et al. (De Ryck et al., 1989), in a modification of Yolkkonen et al. (Jolkkonen et al., 2000). This test consists of 7 exercises in which the reaction of the front and hind limbs to tactile and proprioceptive stimulation is determined. Tasks were evaluated on a 3-point scale: 2 points — normal reaction; 1 point - the reaction is weakened or with a delay of ≥2 s; 0 points - no reaction. The maximum number of points was 14. Rats were tamed for several days and then tested 1 day before the operation (“baseline”) and 1, 2, 3, 5, and 6 days after the operation. Tested right and left limbs. Forelimb-placing test. The test was carried out according to the technique described in detail in (Jolkkonen et al., 2003).

Statistical data processing. Statistical analysis was performed using the program Statistica 6.0 (StatSoft, USA) for Windows. The normality of the distribution of the trait in the sample was evaluated by the Shapiro-Wilk W test. For comparison, the indices obtained in the Limb-placing and Forelimb-placing tests were performed using ANOVA analysis with Kruskal-Wallis criteria. For intragroup comparisons, the Mann-Whitney U test with Bonferroni correction was used.

The most clear and indicative results were obtained in the Limb Placing test. To reduce dispersion, the ratio of scores for the right and left limbs was used as an indicator. From the data obtained in the Forelimb Placing and Limb Placing tests, it is clearly seen that in the first days of testing (days 1 and 2 - 1 and 2 days after surgery) as a result of ischemic damage to the sensorimotor region of the left hemisphere cortex, selective deterioration of neurological parameters for the right limbs occurs, but starting from the 2nd day (2 days after the operation), a gradual restoration of neurological indicators was observed in all experimental groups. Almost complete recovery of neurological parameters was observed already on the 7th day after the operation. At the same time, in false-operated animals, the disturbances were insignificant.

We studied the effect of heptapeptide sequences of Met-Glu-His-Phe-Pro-Gly-Pro at a dose of 15 μg / kg in saline, antioxidants, and a mixture of the heptapeptide with antioxidants on the change in neurological parameters after focal ischemia of the sensorimotor region in the Limb Placing test. The rats were given 8 intraperitoneal injections of these drugs within seven days after surgery. The results of this experiment are presented in table 4. As can be seen from this table, a statistically significant improvement in neurological parameters was established compared with the control 2, 3, and 5 days after photochemically induced thrombosis (Table 4 in groups 5-7). In this case, a pronounced synergistic effect was observed when using drugs that are a mixture of heptapeptide and antioxidants (groups 5-7), compared with control groups, with the introduction of only heptapeptide sequences of Met-Glu-His-Phe-Pro-Gly-Pro or only antioxidants . Injection forms, made in the form of lyophilized powder, containing the heptapeptide sequence Met-Glu-His-Phe-Pro-Gly-Pro in combination with an antioxidant (α-lipoic acid - composition 6, ascorbic acid - composition 5 and α-tocopherol acetate - composition 7), show significant effectiveness in experimental treatment of stroke, while the heptapeptide sequences Met-Glu-His-Phe-Pro-Gly-Pro (15 μg / kg) and antioxidants (ascorbic acid, α-lipoic acid, α-tocopherol acetate), taken separately, do not lead to a significant improvement in neurovol ogichesky indicators in comparison with control. Thus, this experiment confirms the presence of a synergistic effect.

Example 13. The study of the stability of the heptapeptide sequence of Met-Glu-His-Phe-Pro-Gly-Pro with the addition of an antioxidant in various injection forms compared with a solution of the heptapeptide of the sequence Met-Glu-His-Phe-Pro-Gly-Pro, prepared in physiological sodium chloride solution.

To experimentally study the stability of the proposed injectable dosage forms of the Met-Glu-His-Phe-Pro-Gly-Pro preparation (with the addition of an antioxidant) and determine the shelf life, model mixtures of several formulations of the preparation were made (Table 5). As a comparison drug used the drug "Semax" for intranasal use of 1% solution (manufacturer of the IPPC "Peptogen") and 1% solution of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro in an isotonic 0.9% solution sodium chloride, prepared without the addition of antioxidants and stabilizing additives. Samples were stored in a cool place protected from light and relative humidity (60 ± 5)% during the observation period of 3 years. The results of stability studies under natural storage conditions are presented in Table 5.

Table 5
The results of a study of the stability of the heptapeptide sequence of Met-Glu-His-Phe-Pro-Gly-Pro in the proposed injectable dosage forms (lyophilized powder, dry powder, emulsion, solution for intramuscular or intravenous administration) The composition of the drug The concentration of the active substance (heptapeptide sequence Met-Glu-His-Phe-Pro-Gly-Pro) relative to the freshly prepared solution, determined by HPLC,% 1 month 6 months 12 months 18 months 24 months 36 months Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 5 wt.%, Ascorbic acid 25 wt.%, Α-lipoic acid 5 wt.%, Glycine 65 wt.% (Lyophilized powder, example 7) 99.68 ± 0.05 99.50 ± 0.12 99.31 ± 0.09 98.56 ± 0.04 98.51 ± 0.11 97.83 ± 0.15 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.17 wt.%, Ascorbic acid 2.34 wt.%, Sodium chloride 1.40 wt.%, Sodium hydrogen phosphate 1.40 wt.%, Sodium dihydrogen phosphate 0.70 wt.%, Beckons 93.02 wt.% (Lyophilized powder, example 2) 98.84 ± 0.21 98.42 ± 0.18 98.02 ± 0.14 97.90 ± 0.12 97.31 ± 0.09 97.25 ± 0.11 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.35 wt.%, Quercetin 0.10 wt.%, Polyvinylpyrrolidone 0.15 wt.%, Trilon-B 0.10 wt.%, Glycine 0, 50 wt.%, Water for injection up to 1 ml (solution for injection, example 3) 99.26 ± 0.25 99.16 ± 0.18 98.82 ± 0.05 98.61 ± 0.14 98.51 ± 0.11 98.19 ± 0.06 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1 wt.%, Α-tocopherol acetate 1.0 wt.%, Polysorbate-80 0.5 wt.%, Nipazole 0.05 wt.%, water for injection 12.5 wt.%, olive oil up to 2 ml (85.85 wt.%) (solution for injection, example 6) 99.52 ± 0.03 99.53 ± 0.07 99.40 ± 0.12 99.14 ± 0.05 99.02 ± 0.05 98.95 ± 0.13 Semax for intranasal use 1% solution (Peptogen) 98.01 ± 0.05 97.16 ± 0.09 95.31 ± 0.17 94.82 ± 0.08 87.68 ± 0.12 78.01 ± 0.19 Heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 1.0 wt.%, Sodium chloride solution isotonic 0.9% 1 ml. 97.95 ± 0.16 97.06 ± 0.04 95.22 ± 0.09 92.15 ± 0.23 83.41 ± 0.22 62.27 ± 0.16

The results of studies on the stability of the heptapeptide of the Met-Glu-His-Phe-Pro-Gly-Pro sequence in injectable dosage forms with the addition of an antioxidant (lyophilized powder, dry powder, emulsion, solution for intramuscular or intravenous administration) confirm that the introduction of the antioxidant into the composition of the drug significantly stabilizes the peptide molecule and contributes to an increase in shelf life compared to a drug in which there are no excipients with an antioxidant effect. It was found that the most stable dosage forms of the drug, made in the form of lyophilized powder, dry powder and oil solution for injection.

In addition, the proposed composition provides the ability to store the drug at room temperature, can withstand freezing / thawing at least 4 times, remains active for 7 days at a temperature of + 35 ° C, and also after prolonged storage (1 year) at a low temperature of -20 ° From and on for 2 years. The addition of auxiliary substances contributes to an easier release of the active substance and ensuring its high bioavailability.

Claims (11)

1. Injection dosage form based on a heptapeptide of the Met-Glu-His-Phe-Pro-Gly-Pro sequence for the treatment of acute ischemic stroke and traumatic brain injury, characterized in that it contains a therapeutically effective amount of Met-Glu- heptapeptide as an active ingredient His-Phe-Pro-Gly-Pro and at least one substance selected from the group of antioxidants including β-carotene, retinol acetate, α-tocopherol acetate, ascorbic acid, lutein, quercetin, rutin (vitamin P), dihydroquercetin , butylhydroxytoluene, α-lipoic ki lot, L-carnitine, succinic acid, and, optionally, at least one excipient selected from the group of stabilizers, prolongers, buffering agents, emulsifiers, solubilizers, solvents, fillers, preservatives and other excipients permitted for medical use, in the following ratio of components, wt.%:
heptapeptide 0,0002-99,99998 antioxidant 0.00002-99.9998 2. The injection dosage form according to claim 1, characterized in that it further comprises at least one excipient selected from the group consisting of stabilizers, prolongers, buffering additives, emulsifiers, solubilizers, solvents, fillers, preservatives and other auxiliary substances approved for medical use, while containing as stabilizer at least one substance selected from the group consisting of Trilon B, sodium metabisulfite, sodium thiosulfate, glycine, arginine, histide in, lysine or their physiologically acceptable salts, for example hydrochloride, sulfate, acetate, glutamate, aspartate and maleate, as a prolongator, at least one substance selected from the group consisting of polyvinylpyrrolidone with a molecular weight of 10-60 kDa, molecular weight dextran weighing 10-100 kDa, for example polyglucin or reopoliglukin, polyvinyl alcohol, sodium carboxymethyl cellulose, as a buffering additive - at least one substance selected from the group consisting of sodium chloride, sodium / potassium hydro- and / or dihydrogen phosphate, sodium ia or ammonium acetate, as an emulsifier-solubilizer - at least one substance selected from the group comprising soya lecithin for injection, polysorbate-20, polysorbate-60, polysorbate-80, span-20, span-40, span 60, span-85, sodium dodecyl sulfate, as a solvent, at least one substance selected from the group consisting of water for injection, olive oil, peach oil, sunflower oil, at least one substance selected as a filler from the group including sorbitol, mannitol, xylitol, lactose, sucrose, dext ozu, a copolymer of D, L-lactic and glycolic acids, as a preservative - at least one substance selected from the group consisting of chlorobutanol hydrate, ethyl alcohol, benzyl alcohol, phenol, chlorocresol, benzoic acid, sorbic acid, merthiolate, nipagin, nipazole, benzalkonium chloride, benzethonium chloride, cetylpyridinium chloride, dimethyldodecylbenzylammonium chloride. 3. An injection dosage form according to any one of claims 1 and 2, characterized in that it can be in the form of a lyophilized powder, dry powder, emulsion, solution for intramuscular or intravenous administration, or an injection solution for infusion. 4. Injection dosage form according to claim 2, characterized in that it contains in each ampoule or vial, wt.%:
heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1-5.0 routine 0.05-5.0 glycine 0.00-5.0 lactose up to 100.0
and made in the form of dry powder. 5. Injection dosage form according to claim 2, characterized in that it contains in each ampoule or vial, wt.%:
heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1-5.0 ascorbic acid 0.01-10.0 sodium chloride 0.02-2.0 sodium hydrogen phosphate 0.01-1.00 sodium dihydrogen phosphate 0.01-0.50 beckons up to 100.0
and made in the form of dry powder. 6. The injection dosage form according to claim 2, characterized in that it contains in each ampoule or vial, wt.%:
heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.1-2.0 quercetin 0.005-1.0 polyvinylpyrrolidone 0.005-5.0 trilon-B 0.00-1.0 glycine 0.001-5.0 water for injection up to 100.0
and made in the form of a solution. 7. Injection dosage form according to claim 2, characterized in that it contains in each bottle or container for infusion solutions, wt.%:
heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.0002-0.004 ascorbic acid 0.00002-0.1 succinic acid 0.00002-0.06 lysine 0.00-0.006 molecular weight dextran 10-100 kDa 0,0-6,0 0.9% sodium chloride solution isotonic or glucose solution 5-10% to 100.0
and made in the form of a solution for infusion. 8. Injectable dosage form according to claim 2, characterized in that it contains in each ampoule or vial, wt.%:
heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.05-1.0 α-tocopherol acetate 0.0025-2.5 polysorbate-80 0.005-0.5 nipazole 0.0025-0.005 water for injection 5.0-50.0 olive oil up to 100.0
and made in the form of an emulsion. 9. Injection dosage form according to claim 2, characterized in that it contains in each ampoule or vial, wt.%:
heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro 0.50-15.0 ascorbic acid 0.25-35.0 α-lipoic acid 0.25-17.5 glycine up to 100.0
and made in the form of lyophilized powder. 10. A method of preparing a dosage form for injection, described in any one of claims 1 to 9, comprising mixing the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro with an antioxidant and the corresponding at least one excipient and, if necessary, sterilization of the injectable dosage form; pouring the solution into ampoules, vials or containers for infusion solutions; if necessary, drying solutions in a sublimation unit; sealing of ampoules and containers for infusion solutions or corking of finished product bottles. 11. The use of the injectable dosage form of the heptapeptide Met-Glu-His-Phe-Pro-Gly-Pro, characterized in any one of claims 1 to 9 for the treatment of acute ischemic stroke and traumatic brain injury.

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