RU2330680C2 - Injection dosage form for acute stroke treatment, method of production and application thereof - Google Patents

Abstract

FIELD: medicine; pharmacology.

SUBSTANCE: injection dosage form for acute stroke treatment includes noopeot in therapeutically effective amount as reactant, and as adjuvants, at least one selected from group including lactose, saccharose, polyvinyl alcohol, polyvinylpyrrolidone, sorbite, mannitol, glycine, aqueous solutions, fatty oils, ethyl oleate, glycerine, ethyl alcohol, propylene glycol, polyethylene oxide 400, benzyl benzoate, benzyl alcohol and other adjuvants allowed for medical application. Injection dosage form meets all requirements of State Pharmacopeia, XI edition. Medical product Noopept (N-phenyl acetyl-L- prolyl glycine ethyl ether) as injection dosage form provides high nootropic and neuroprotective activity applied to models of extensive ischemic stroke and hemorrhagic stroke.

EFFECT: stabile storage ability and high bioavailability.

8 cl, 1 tbl, 2 ex

Description

The invention relates to medicine, in particular to pharmacology and pharmaceuticals, and for the injection form of the drug NOOPEPT for the treatment of the early stages of stroke.

Stroke is a vascular lesion of the brain caused by either circulatory failure (ischemic stroke) or cerebral hemorrhage (hemorrhagic stroke). According to world statistics, annually a stroke affects 6 million people, and if its prevalence in the world is 460-560 cases per 100 thousand people, then in Russia this figure is 1050 cases. Stroke ranks third among the causes of death in the world and second in Russia. The frequency of development of ischemic strokes is 70-80%, hemorrhagic strokes account for 20-30%. During the first month of the disease, 15-30% of patients with ischemic strokes and 30 to 70% of patients with hemorrhagic stroke die.

The success of treatment of stroke patients is determined mainly by the early dates of initiation of therapy, the strategy of which differs significantly in ischemic and hemorrhagic stroke. Therapy of ischemic stroke is aimed at restoring patency of the artery, preventing further buildup of the blood clot, and then at preventing the death of neurons in the penumbra zone (penumbra) using neuroprotectors. With hemorrhagic stroke, the use of neuroprotectors is also advisable, but drugs that prevent thrombosis are contraindicated. However, differential diagnosis of these two forms at the earliest stages of the disease is difficult, and in more than 25% of cases it was completely impossible. These facts dictate the need to develop such drugs that can be effective in both forms of strokes. Of the currently used drugs, it is possible to point out calcium channel blockers, however, a significant drawback of these drugs is the presence of cardiovascular effects, which lead to "brain robbery."

The drug NOOPEPT - N-phenylacetyl-L-prolylglycine ethyl ester (RF patent No. 2119496, published in BI No. 27, 1998), which belongs to a new group of compounds, has various types of psychotropic activity, including antiamnestic, antihypoxic and anorexigenic action.

Previously, a pharmaceutical composition was developed containing effective doses of NOOEPEPTA for oral use, mainly in the form of tablets, which can be used to treat patients suffering from mental decline caused by brain injury, aging, prenatal hypoxia or alcoholism, as well as weakening the manifestation of the withdrawal syndrome of benzodiazepine tranquilizers (Application for the grant of a patent of the Russian Federation for invention No. 2004123357). Currently, as a nootropic drug (oral dosage form, 5 mg tablets), NOOEPT passed the I and IIa phase of clinical trials that showed the safety of the drug and its effectiveness for the treatment of moderate cognitive impairment of cerebrovascular and post-traumatic origin.

The effectiveness of the NOOPEPT substance on a model of small ischemic stroke caused by photoinduced thrombosis, in which the volume of damage to the cerebral cortex did not exceed 2% of the volume of the cerebral hemisphere, was described (Ostrovskaya RU, Romanova GA, Gudasheva TA et al., Behav. Pharmacol., 1999, 10, 549-553). A decrease in the volume of the affected area was revealed and it was shown that the neuroprotective effect of the NOOPEPT preparation is based on the ability to prevent the development of metabolic disorders such as the accumulation of free radical forms of oxygen, intracellular calcium and excess glutamate. These fundamentally important properties of the drug in combination with the anticoagulant and fibrinolytic activity revealed in it in the absence of antiplatelet action (i.e., effect on primary hemostasis) exclude the development of increased bleeding, which is dangerous in hemorrhagic stroke, when it is used.

The essence of the invention lies in the development of an injectable dosage form of the drug NOOPEPT, which, when administered parenterally on models of massive ischemic stroke (lesion volume is 18-20% of the total volume of the entire hemisphere of the brain) and hemorrhagic stroke, provides a quick onset of effect and the possibility of use in those patients in whom due to their condition, the use of an oral form of the drug is impossible.

An injectable dosage form in the form of a pharmaceutical composition was developed containing NOOPEPT as an active ingredient, as well as a sufficient amount of excipients, which made it possible to obtain a stable dosage form that easily releases the active substance and ensures its high bioavailability.

NOOPEPT, on the one hand, is slightly soluble in water, and for its dissolution it is necessary to use excipients that increase its solubility, and on the other hand, being an ester, it undergoes hydrolysis in contact with water, and to obtain a stable injectable dosage form reduce the rate of its destruction.

These problems were solved by using aqueous solutions, fatty oils, ethyl oleate, ethyl alcohol, glycerol, propylene glycol, polyethylene oxide 400, benzyl sprite, benzyl benzoate.

The stability of the injectable dosage form of NOOPEPTA was achieved by the manufacture of dry solid powders, porous masses obtained in the form of dry powders, lyophilized powders containing therapeutically effective amounts of the drug and auxiliary excipients, which include lactose, sucrose, polyvinyl alcohol, polyvinylpyrrolidone, sorbitol , mannitol, glycine.

As a filler in the composition, one or more compounds can be used that are capable of providing an injection mass of good quality. The use of filler in an amount close to the lower limit of the mass range of the filler is desirable.

For sterilization of the drug due to the thermolability of its substance, the sterile filtration method was used.

The following are possible formulations for the preparation of an injectable dosage form of NOOPEPTA.

1. Obtaining dry powder powders.

To obtain a stable injectable dosage form of NOOPEPTA by dry grinding, triturations of Noopept and auxiliary substances are prepared in order to increase the solubility of the substance and the stability of the solution. Lactose, sucrose, mannitol, glycine and other excipients permitted for medical use can be used as a filler in the composition for obtaining dry powder. The contents of the ampoule are dissolved in water for injection or in physiological saline immediately before injection.

The pharmaceutical composition is made in solid form, in appearance it is an amorphous powder in an ampoule or vial. An ampoule or vial contains:

Noopept - 0.0005-0.02 g.

Lactose - 0.005-0.2 g.

2. Obtaining an injection solution based on non-aqueous solvents.

To exclude the contact of NOOPEPTA with water, the following substances and their compositions can be used as a solvent for injection: fatty oils, ethyl oleate, glycerin, ethyl alcohol, propylene glycol, polyethylene oxide 400, benzyl benzoate, benzyl alcohol and other solvents approved for medical use. As a solvent in the composition of the present invention, one or more compounds can be used that are capable of providing an injection solution of appropriate quality.

The pharmaceutical composition is in liquid form, in appearance it is a viscous liquid in an ampoule. The ampoule contains:

Noopept - 0.0005-0.02 g.

Olive oil - up to 1 ml.

3. Obtaining a lyophilisate for the preparation of an injection solution.

To obtain a storage-stable and readily soluble lyophilisate for injection, auxiliary excipients are used, which include lactose, sucrose, polyvinyl alcohol, polyvinylpyrrolidone, sorbitol, mannitol, glycine and other auxiliary substances approved for medical use.

As filler in the composition of the present invention can be used one or more compounds capable of providing a lyophilized mass of the proper quality.

As a solvent for obtaining a solution for freeze-drying, purified water is used, a mixture of purified water with ethyl alcohol, tert-butanol.

The following are possible formulations for preparing a lyophilized dosage form of NOOPEPTA.

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

Noopept - 0.0005-0.02 g.

Mannitol - 0.005-0.2 g.

The pharmaceutical composition based on NOOPEPTA can be obtained by any known lyophilization method. One of these methods involves the following steps:

1) preparation of a solution containing noopept and excipients;

2) sterile filtration of the resulting solution;

3) pouring the solution into ampoules or vials;

4) drying the solution in a sublimation unit with preliminary freezing in a freezer or in a chamber of a sublimation unit;

5) sealing of ampoules, corking of bottles with the finished product.

The following methods are used to sterilize the drug: 1) aseptic preparation; 2) thermal sterilization; 3) sterile filtration.

The following are examples illustrating the effectiveness of NOOEPEPT on a model of massive cortical ischemia and hemorrhagic stroke.

Example No. 1. The study of the effectiveness of the injectable dosage form of NOOPEPTA on the model of massive cortical ischemia.

The experiments were performed on white outbred male rats weighing 350-450 g, anesthetized with chloral hydrate (400 mg / kg, ip), which were ligated the left carotid artery, and then under the microscope (magnification 14.0 × 3.3), the middle cerebral artery was occlusion (OSMA ) proximal to the place of its bifurcation on the frontal and parietal branches. After the operation, the animals were randomly divided into 2 groups. The animals of the experimental group were administered NOOEPEPT in the form of an injectable dosage form for parenteral use. The drug was administered intraperitoneally after 30 minutes, then after 2, 24 and 48 hours after surgery. The animals of the control group were injected with saline (0.9% sodium chloride) at the same time. 72 hours after OSMA, the animals were decapitated (anesthesia with chloral hydrate 800 mg / kg, ip). Five front sections, 2.5 mm thick, were prepared from frozen at -20 ° C, which were stained with a solution of 2,3,5-triphenyltetrazolium chloride (TFC). Using the Auc1 morphometric program, planimetry was performed, the volume of the lesion zone and the percentage of the lesion zone volume relative to the ipsilateral hemisphere volume were determined. To compare the severity of changes in different sections, a conditional scale was used, in which a lesion of 75% of the hemisphere was rated as 2.5 points, 60% - 2 points, 45% - 1.5 points, 20% - 1 point, 5% - 0.5 points, 0% - 0 points.

This operation creates an extensive ischemic lesion of the cerebral cortex, localized within the front-parietal and dorso-lateral zones. In control animals treated with saline, the total lesion volume is 18.65 ± 2.65%. The maximum ischemic damage occurs at the level of the 2nd and 3rd sections (table).

Table The effect of the injectable dosage form of NOOPEPTA on the severity of ischemic lesions of the cerebral cortex (score sections) one 2 3 four 5 Fiz.rr (n = 8) 0.5 ± 0.11 1.44 ± 0.2 1.38 ± 0.22 0.69 ± 0.15 0.19 ± 0.1 Noopept 0.5 mg / kg (n = 9) 0.06 ± 0.01 * 1.06 ± 0.1 * 0.8 ± 0.2 * 0.63 ± 0.12 0.19 ± 0.1 * p <0.05

NOOEPTOM therapy leads to a significant reduction in damage at the level of the first three sections in the absence of significant changes at the level of the 4th and 5th sections (table). The total lesion area decreased to 12.21 ± 2.65%. From comparison with the group of control animals, it follows that NOOPEPT, applied after ischemic exposure, reduces the total lesion volume by 34.5% (p <0.05). The totality of the obtained data indicates that the studied drug, introduced after the implementation of the ischemic effect, significantly reduces the severity of morphological changes in the cerebral cortex due to coagulation of the midbrain artery in combination with ligation of the ipsilateral carotid artery.

In a separate series of experiments, animals were trained in the test of conditioned passive avoidance reflex (passive avoidance reaction), developed in the apparatus "Lafayette Instrument Co", USA), consisting of a brightly lit hanging platform connected to a dark chamber with an electrified floor. Due to the congenital mink reflex, the rat entered the dark chamber during training, where it received electric pain irritation. The memory was evaluated 24 hours after training, recording the number of animals that did not enter the dark chamber for 180 s, and for those who entered it, the latent time of entry. As shown earlier in numerous control experiments on intact rats, they usually do not enter the danger chamber or enter at the very end of the test (average latent period 158 ± 12.1 s). In ischemic animals treated with saline, the latency of approach is 20 ± 6.1 s (most animals did not remember the irritation caused the day before and quickly entered a dangerous chamber). The group of ischemic animals treated with noopept showed an increase in the latent period to 67 ± 9.3 s, which indicates an improvement in memory.

Example No. 2. Study of the effectiveness of the injectable dosage form of NOOPEPTA on the model of hemorrhagic stroke.

In experiments on male outbred white rats anesthetized with nembutal (40 mg / kg, w / m), the simulation of intracerebral post-traumatic hematoma (HI) was performed by mechanical destruction of brain tissue in the area of the internal capsule and introduction of blood taken from under the tongue into the site of damage (0, 02-0.03 ml). All animals were randomly divided into 4 groups (in each of 10-15 animals): 1) intact animals that were injected with saline; 2) animals with HI, which were injected with saline; 3) a group of animals with HI, which was introduced NOOPEPT at a dose of 0.5 mg / kg once; 4) animals with GI treated with NOOPEPT repeatedly for 14 days in the same dose. Saline or injectable dosage form of NOOPEPTA at a dose of 0.5 mg / kg was administered intraperitoneally, with the first injection in animals of the 3rd group once, and in animals of the 4th group, the first injection was also carried out after 4 hours, the second after 8 hours, on the second and third days, two injections were carried out, from the 4th to the 14th day - one injection per day.

Analysis of the neurological status of the surviving animals showed that the presence of weak neurological disorders was detected in 85% of animals in group 2 (GI + saline) and only in 24% of rats with GI treated with NOOEPT. The drug also reduced the incidence of severe neurological symptoms: if in the group by the 7th, 14th days this indicator was 65 and 70%, NOOPEPT reliably attenuated the neurological deficit, while its effect was more pronounced on the 7th, 14th day, this indicator decreased to 20 and 18%, respectively.

When registering the behavior of animals in the passive avoidance reaction test, the nootropic effect of NOOPEPT is shown. For example, if in the control group, when the reflex was reproduced on the 7th day after the operation, 92% of the animals remembered the pain stimulus applied in the dark chamber and did not enter it, and in animals after GI treated with saline, only 30% remembered this and did not enter into it, NOOPEPT, administered once at a dose of 0.5 mg / kg and reused in the same dose, significantly increased the number of trained animals to 67% and 60%. Injection drug significantly increased the latent period of entry into the dark chamber. If in the control group this indicator was 160 ± 18 s, then in the group of animals with GI treated with saline, it decreased to 60 ± 15 s, and in animals with GI treated with NOOPEPT, there was a significant increase in the latent period (up to 135 ± 15 s), which indicates a pronounced antiamnestic effect of the drug.

In the open field test, it was shown that in the group of rats with HI treated with saline, there was a significant decrease in horizontal and vertical activity, the number of examinations of holes. NOOPEPT, when administered at a dose of 0.5 mg / kg, both once and for 14 days, significantly increased activity. When studying the effect of the injectable form of NOOPEPTA on the coordination of movements and muscle relaxation of animals in tests of a rotating rod and horizontal crossbar, violations were revealed in the groups of stroke rats. An injection drug, administered at a dose of 0.5 mg / kg once, halved the number of animals with impaired coordination of movements on the 7th and 14th day after surgery, and with chronic administration according to the scheme for 14 days it was statistically significant ( up to 3 times) reduced the number of animals with impaired coordination of movements throughout the entire observation period.

It is important to emphasize that both in the case of ischemic and in the case of hemorrhagic strokes, the injectable dosage form of the NOOPEPT preparation was administered after the damaging effect, while if the interval was 30 minutes on the model of ischemia, then in conditions of hemorrhagic strokes the first injection was carried out after 4 hours. This fact indicates a fairly wide "therapeutic window" for the studied drug.

Claims (8)

1. Injectable dosage form based on noopept for the treatment of acute stroke, comprising as the active substance a therapeutically effective amount of noopept, and as excipients, at least one selected from the group comprising lactose, sucrose, polyvinyl alcohol, polyvinylpyrrolidone, sorbitol , mannitol, glycine, aqueous solutions, fatty oils, ethyl oleate, glycerin, ethyl alcohol, propylene glycol, polyethylene oxide 400, benzyl benzoate, benzyl alcohol and other excipients permitted to me ditsinsky application. 2. The injection dosage form according to claim 1, characterized in that it contains in each ampoule or vial 0.0005-0.020 g of noopept. 3. The injection dosage form according to claim 1, in the form of a dry powder containing Noopept 0.0005-0.020 g Lactose 0.005-0.2 g 4. Injectable dosage form according to claim 1, containing Noopept 0.0005-0.020 g Olive oil up to 1 ml 5. Injectable dosage form according to claim 1 in the form of a lyophilized powder containing Noopept 0.0005-0.020 g Mannitol 0.005-0.2 g 6. A method of preparing an injectable dosage form, characterized in any one of claims 1 to 5, comprising mixing noopept and appropriate auxiliary substances, and, if necessary, sterilizing the injectable dosage form; pouring the solution into ampoules or vials; drying solutions in a sublimation unit; sealing ampoules or corking finished product bottles. 7. The use of an injectable dosage form, characterized in any one of claims 1 to 5, for the treatment of acute stroke. 8. The use according to claim 7 for emergency treatment of ischemic or hemorrhagic forms of stroke.