RU2304977C2 - Method for adjusting genome instability in children suffering from infantile cerebral paralysis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves applying mono therapy with Cerebrolysin intramuscularly introduced once a day in the morning at a daily dose of 0.1 ml/kg of weight as 20 days long course or Encephabol to be taken 3 times a day in 30 min after having taken meals. Encephabol is given at a dose of 50 mg in 2-5 years old children cases or at a dose of 100 mg in 6-10 years old children cases. Male children take Encephabol in not longer than 10 days long course. Female children take Encephabol in a course being longer than 10 days.

EFFECT: enhanced effectiveness of genome instability adjustment.

6 dwg

Description

The present invention relates to medicine, namely to children's neurology.

A known method for the medical treatment of cerebral palsy (cerebral palsy), including various drugs, such as piracetam, cerebrolysin, encephabol, actovegin, cavinton and others with specific nootropic and neurotrophic effects. These funds have been used since the first days of treatment of perinatal pathology, the cerebral palsy of which is a serious outcome. In the absence of contraindications, the drugs are used in various combinations among themselves with repeated courses (2-3 times a year).

There is a method of rehabilitation treatment using physiotherapeutic procedures (diadynamic and sinusoidally modulated currents, drug electrophoresis, microwave resonance therapy, mud therapy, etc.), physiotherapy exercises (methodology according to K.A.Semenova, Koysiotherapy according to Voight, etc.) and massage (general, acupressure, segmental). The courses of treatment are repeated, repeatedly throughout the year with small interruptions. One course of treatment, depending on the type, includes 8-20 procedures.

The aim of the study is to experimentally evaluate the antimutagenic and antioxidant activity of some neurotropic drugs used in the treatment of the chronically residual stage of cerebral palsy, select the most and least active substances from the results, and in clinical studies evaluate their ability to normalize the processes that destabilize the gene in patients with cerebral palsy.

The disadvantage of the prototype is the lack of a differentiated approach to the prescription of drugs, which, along with a specific neurotropic pharmacological activity, have an antimutagenic effect, as well as complex therapy of cerebral palsy include drugs with a clastogenic and / or prooxidant effect, which enhance the destabilization of the genome.

The method is as follows: in the experiment, the antimutagenic and antioxidant activity of some neurotropic drugs, the most widely used in complex therapy of the chronically residual stage of cerebral palsy, was evaluated. Based on the results, the most and least active drugs were selected. Then, in clinical trials, their ability to normalize the processes destabilizing the genome in patients with cerebral palsy, i.e. carried out pharmacological protection of the genome in patients with cerebral palsy.

For experimental and clinical studies, neurotropic agents used in the treatment of the chronically residual stage of cerebral palsy were used: cerebrolysin (Ebewe), encephabol (Nycomed), nootropil (Pliva dd) and its domestic analogue, piracetam (Moskhimpharmpreparat), cavinton (Gedeon Richter), acteg Nycomed).

1. Antioxidant and antiradical activity of neurotropic drugs

Nootropilum possessed the highest antioxidant activity under such conditions (C ₅₀ = 40 μl) (Table 1). In order to attenuate the effect, actovegin (C ₅₀ = 43 μl), piracetam (65 μl), cerebrolysin (82 μl), cavinton (above 100 μl) follow. Encephabol, in contrast, enhanced the formation of malondialdehyde, i.e. possessed prooxidant activity. Thus, five of the six drugs studied reduced the LPO intensity in the model system by 50% in the range from 40 to 80 μl / ml of the drug in the sample.

Only Cerebrolysin (C ₅₀ = 60 μl / ml) and Actovegin (C ₅₀ exceeded 100 μl and the effect was weak) possessed superoxide dismutase activity (SOD activity). The remaining drugs in this model system enhanced chemiluminescence, i.e. showed prooxidant activity.

The most active drug in its ability to directly react with nitric oxide (NO) was cerebrolysin (C ₅₀ = 4.25 μl / ml). Actovegin (C ₅₀ = 24 μl / ml) and encephabol (C ₅₀ = 33.7 μl / ml) showed significantly less activity against nitric oxide. Piracetam, Nootropil, Cavinton preparations at concentrations up to 150 μl / ml did not have an effect in the selected system.

Comparing data on the activity of drugs in various model systems, cerebrolysin can be considered the most effective antioxidant and antiradical drug. Of the 6 drugs studied, cerebrolysin has the highest ability to inhibit the activity of superoxide anion radical (SOD) and nitrogen monoxide (NO). Encephabol, the only of 6 drugs, has a pro-oxidizing ability, weakly inhibits nitrogen monoxide and does not have the ability to reduce the activity of superoxide anion radical.

1. The study of anticlastogenic activity of neurotropic drugs

The antimutagenic effect (or anticlastogenic activity) of some neurotropic drugs was studied using ethyl methyl sulfanate, the strongest mutagen that causes 100% mutation (aberration) on the plant medium.

It turned out that only some drugs showed a pronounced ability to suppress the intensity of chemically induced mutagenesis (Table 2). The maximum anticlastogenic activity was shown by cerebrolysin and to a lesser extent cavinton (by 43.0 and 24.6%, respectively). The pronounced antimutagenic effect of cerebrolysin can be associated with the antioxidant properties of the components that are included in its dosage form.

Unlike cerebrolysin, encephabol showed a low anticlastogenic effect: the antimutagenic activity of encephabol was more than 2 times lower than that of cerebrolysin (18.2 and 43.0%, respectively).

Thus, taking into account the results of our experiments on the anticlastogenic, pro- and antioxidant properties of some drugs used in the treatment of cerebral palsy, for clinical trials we selected two drugs from the group of neurotropic drugs: cerebrolysin, the drug with the highest antioxidant and antiradical ability, and Encephabol, which has prooxidant properties, weak antiradical ability and low anticlastogenic effect.

3. Dynamics of the level of micronuclei in polychromatophilic peripheral blood erythrocytes in patients with cerebral palsy under the influence of cerebrolysin and encephabol.

The study on two drugs - cerebrolysin and encephabol after informed consent included 20 patients (9 girls and 11 boys) 2-10 years old with a diagnosis of cerebral palsy, spastic diplegia of moderate severity. All patients were divided into two groups: the first group (6 boys and 4 girls) took cerebrolysin, the second (5 boys and 5 girls) took encephabol. The choice of form — spastic diplegia — is determined by its intermediate position in terms of the level of cytogenetic disturbances in comparison with other clinical forms of cerebral palsy (Table 3).

All patients had a prerequisite for the duration of the experiment was the absence of concomitant somatic diseases. Drugs were not prescribed for children with epilepsy and uncoupled paroxysmal reactions. If it is necessary to use other drugs, along with the above-mentioned nootropic drugs (for example, antipyretic or antitussive drugs, etc.), the patient was excluded from further research. 30 days before monotherapy, patients did not receive medication and physiotherapy, and no instrumental examination was performed.

The effectiveness of monotherapy with cerebrolysin and encephabol was assessed by clinical indicators, the dynamics of the number of red blood cells with micronuclei (EM) and the percentage reduction in the level of EM - antianugen activity. EM level was determined at three points: on the day of monotherapy initiation (before treatment), after 10 and 20 days of treatment.

Cerebrolysin was used IM once a day, in the morning, in a standard daily dose at the rate of 1 ml of the drug per 10 kg of the child's weight (0.1 ml / kg). Encephabol was administered orally at 50 mg from 2 to 5 years and 100 mg from 6 to 10 years 3 times a day 30 minutes after a meal.

As a result of treatment of patients with cerebrolysin and encephabol, positive clinical dynamics were observed: in 9 out of 10 children taking cerebrolysin, and in 8 out of 10 patients taking encephabol. In patients, spastic hypertonicity in the affected limbs decreased, the range of active movements expanded, and oculomotor disturbances decreased. Parents noted that against the background of cerebrolysin therapy in children, interest in the outside world increases, speech and memorization skills improve. When taking encephabol, the characteristics of behavior improve, the general vitality increases, positive changes in the emotional sphere are noted.

In all patients included in the study (main group), the initial EM level (before treatment) was significantly higher than the number of EM in healthy individuals (control group) (p <0.05).

The dynamics of EM in the studied patients of both groups of patients treated with cerebrolysin or encephabol had some features (Table 4).

It turned out that antianeugenic activity in cerebrolysin by the 10th day of therapy was significantly higher than in encephabol (35.6 and 32.7%, respectively; p <0.001). By the 20th day of therapy, the antianeugenic effect of cerebrolysin increased and reached 37.1%. In the group of patients taking encephabol, by the 20th day of therapy there was some negative dynamics in the level of EM and the antimutagenic effect of the drug slightly decreased (up to 31.4%).

This clearly indicates that the drugs have the ability to suppress the intensity of aneugenesis at the level of the whole organism. It is important to emphasize two points.

Firstly, drugs reduced the intensity of aneugenesis not to the physiological norm, but at best by one third (from 31.4 to 37.1%).

Secondly, in the group of children taking cerebrolysin, during therapy, the level of EM gradually decreased and decreased by almost 1.5 times during the 20 days of treatment. In patients taking encephabol, such a clear regression of cytogenetic disturbances did not occur by the end of therapy, on the contrary, there was a tendency for a certain increase in cells with rearrangements.

For a more detailed study of this phenomenon, the dynamics of the number of EMs in boys and girls separately was studied. It turned out that all the girls who were treated with these drugs showed a gradual decrease in the number of EMs (Figs. 1 and 2). In the group of boys who were on encephabol therapy, after the decrease achieved by the 10th day of treatment, the number of EMs increased by the 20th day.

Consequently, in the group of patients taking encephabol, gender differences in the regression of the number of EMs were revealed: in boys, the antianugenic activity by the 20th day of therapy was significantly lower than in girls.

Thus, for the first time in experimental and clinical studies, the antimutagenic, pro- and antiradical activity of neurotropic drugs was evaluated - cerebrolysin, encephabol (pyritinol), nootropil (piracetam), actovegin and cavinton (vinpocetine). In chemical, enzymatic and cellular model systems, only cerebrolysin inhibited the generation of superoxide anion radical, nitric monoxide and end products of degradation of fatty acid chains. Cerebrolysin possessed the greatest anticlastogenic activity in experiments on Crepis capillaries. Encephabol showed the greatest prooxidant effect and low anticlastogenic activity. The modifying effect of the drugs was different: cerebrolysin more than encephabol reduced the intensity of ane- and clastogenesis in patients. The protective effect of the drugs on day 20 of administration was more effective in the group of girls.

FINDINGS

1. In experimental studies, drugs from the group of neurotropic drugs showed varying degrees of antimutagenic and antioxidant activity. The most active in the studied properties was cerebrolysin.

2. Encephabol showed the least antimutagenic activity, and antioxidant properties were practically absent (prooxidant).

3. For pharmacological correction of damage to the genetic apparatus in patients with cerebral palsy, cerebrolysin is recommended (at a dose of 0.1 ml per kg of body weight daily in the morning of m / m for 20 days), in which, along with specific pharmacological activity, an antimutagenic component is also expressed.

4. For boys with cerebral palsy, encephabol is recommended for a course of no more than 10 days by mouth at a dose of 50 mg from 2 to 5 years and 100 mg from 6 to 10 years 3 times a day 30 minutes after eating.

Information sources

1. Zavadenko N.N. Nootropic drugs in the practice of a pediatrician and pediatric neurologist. M., 2003, 23 p.

2. Voronina T.A., Seredenin S.B. Nootropic drugs, achievements and prospects // Experiment. and wedge. Pharmacology, 1998, No. 4, pp. 3-9.

3. Gonzalez M.E., Francis L., Castelleno O. 1998. System antioxidant activity of Cerebrolyzin // J. Neural Trasm. Vol. 52. P.333-341.

4. Schmid W. The micronucleus test. // Mutat. Res. - 1975 .-- Vol.31. - N 1. - P.9-15.

Table 1. The results of the study of antioxidant and antiradical activity of neurotropic drugs. The method of correction of instability Drug activity Drug name antioxidant antiradical FLOOR (%) SOD (%) NO (%) Encephabol (pyritnol) 0 (in this system, the drug has prooxidative activity) Proosidant C ₅₀ = 33.7 μl / ml Piracetam C ₅₀ = 65 μl / ml Prooxidant No effect
(C _max = 150 μl / ml) Nootropilum (Piracetam) C ₅₀ = 40 μl / ml Prooxidant No effect
(C _max = 150 μl / ml) Actovegin C ₅₀ = 43 μl / ml The effect is weak (C ₅₀ = 100 μl / ml) C ₅₀ = 24 μl / ml Cavinton (vinpocetine) Prooxidant Prooxidant No effect
(C _max = 150 μl / ml) Cerebrolysin C ₅₀ = 8C μl / ml Pronounced effect
(C ₅₀ = 60 μl / ml) C ₅₀ = 4.25 μl / ml Table 2. Anticlastogenic effect of neurotropic drugs Instability correction method ... The name of the drug Anticlastogenic activity (%) Nootropil 14.5 Actovegin 6.9 Piracetam 14.7 Cerebrolysin 43.0 Cavinton 24.6 Encephabol 18.2 Table 3. The level of red blood cells with micronuclei in patients with various clinical forms of cerebral palsy Forms of cerebral palsy The average number of micronuclei Hemiparetic 0.1946 Atonic-astatic 0.2107 Spastic diplegia 0.2114 Hyperkinetic 0.2149 Double hemiplegia 0.2238 Total 0.2111 Healthy 0,0787 Table 4. Antianeugenic activity of cerebrolysin and encephabol preparations during monotherapy in patients with cerebral palsy A drug EM level (%) during therapy Anti-antigenic activity of the drug (%) Source by day 10 by day 20 by day 10 by day 20 Cerebrolysin 0.205 0.132 0.129 ^*
35.6 ^{** *}
37.1 ^** Encephabol 0.296 0.199 0,203 32.7 ^* 31.4 ^* Note: differences in values are significant (p <0.001):
^* - vertical, ^** - horizontal

Claims (1)

A method for correcting genome instability in children with cerebral palsy, which consists in drug treatment of children with elevated levels of red blood cells with micronuclei in the peripheral blood, while the indicated therapy, regardless of the child’s gender, is used intramuscularly with cerebrolysin monotherapy once a day in the morning at a daily dose of 0, 1 ml / kg of body weight for 20 days or with encephabol 3 times a day 30 minutes after meals in the following doses: 50 mg in children from 2 to 5 years old or 100 mg in children from 6 to 10 years old, with male children and encephabol is prescribed for no more than 10 days, and for female children more than 10 days.

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