RU2536234C2 - Neurotropic drug and method of treating structural diseases of nervous system, psychoorganic syndrome and encephalopathies of various origins - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: composition containing an activated-potentiated form of antibodies to the brain-specific S-100 protein, and an activated-potentiated form of antibodies to endothelial NO-synthase as an additional exalting component is administered.

EFFECT: effective treatment of organic diseases of the nervous system by the synergetic neurotropic action of the components.

18 cl, 5 ex, 5 tbl

Description

The invention relates to medicine and can be used for the treatment of organic diseases of the nervous system, including endogenously organic and exogenously organic, for the treatment of the psycho-organic syndrome and encephalopathies of various origins, including those caused by acute and chronic disorders of the cerebral circulation, cranial injuries, neuroinfections, atherosclerotic and neurodegenerative brain lesions.

The psycho-organic syndrome is a symptom complex characterized by a weakening of memory, a decrease in intelligence, and incontinence of affects (the Walter-Buel triad). Asthenic phenomena are often observed. Memory impairment to one degree or another affects all its types. Hypomnesia, in particular dysmnesia, is revealed with the greatest constancy; amnesia and confabulation are possible. The amount of attention is significantly limited, increased distractibility. The quality of perception suffers, in the situation only private details are captured. Orientation worsens, first in the environment, and then in the self. The level of thinking decreases, which is manifested by the impoverishment of concepts and ideas, the weakness of judgments, the inability to adequately assess the situation, its capabilities. The pace of thought processes is slowed down, the torpidity of thinking is combined with a tendency to detail, perseveration. Psycho-organic syndrome (organic psychosyndrome) is a state of mental weakness caused by organic damage to the brain (with vascular diseases of the brain, lesions of the central nervous system, with syphilis, craniocerebral injuries, various intoxications, chronic metabolic disorders, with tumors and abscesses of the brain brain, encephalitis, as well as diseases associated with convulsive seizures). But especially often, the psycho-organic syndrome occurs with atrophic processes of the brain in preschool and senile age (Alzheimer's disease, senile dementia). In its mildest form, the psycho-organic syndrome is an asthenic state with weakness, increased exhaustion, emotional lability, instability of attention, decreased performance. In severe forms of the psycho-organic syndrome, the intellectual-mnestic decrease comes to the first place, reaching the degree of dementia (dementia).

Organic diseases of the nervous system are vascular diseases of the central nervous system (consequences of a stroke, discirculatory encephalopathy), degenerative diseases of the central nervous system, demyelinating diseases of the nervous system, hereditary diseases of the central nervous system, etc.

Parkinson's disease is a chronic, progressive organic disease of the nervous system (neurodegenerative disease), caused by the loss of cells containing dopamine. Degeneration of dopaminergic neurons leads to impaired dopamine synthesis and ultimately to severe motor disorders, impaired coordination of movements and a deterioration in the quality of life of patients.

Encephalopathy is a generic name for non-inflammatory (as opposed to encephalitis) brain diseases. Encephalopathy is congenital and acquired (organic brain damage associated with poisoning, infections, alcoholism, trauma, hypovitaminosis, cerebrovascular disease, lack of vitamin B1). Manifestations: mostly neurosis-like and psychopathic. The treatment of encephalopathy depends on the cause that caused it.

A stroke is an acute cerebrovascular disorder related to cerebrovascular diseases, which is characterized by a sudden (within a few minutes, hours) onset of focal and / or cerebral neurological symptoms, which lasts more than 24 hours or leads to the death of the patient in a shorter period of time due to cerebrovascular pathology. Strokes include cerebral infarction, cerebral hemorrhage, and subarachnoid hemorrhage, which have etiopathogenetic and clinical differences.

It is known from the prior art to treat a psycho-organic syndrome with a neurotropic drug based on an activated-potentiated form of antibodies to the brain-specific protein S-100 (RU 2156621 C1, A61K 39/395, 2000). However, this technical solution may not be effective enough in the treatment of various organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various origins.

The invention is aimed at expanding the therapeutic range of a neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 and increasing the effectiveness of the treatment of organic diseases of the nervous system, including Parkinson's disease, psycho-organic syndrome, including acute cerebrovascular accident (stroke), and various encephalopathies genesis.

The solution to this problem is provided by the fact that a neurotropic drug containing an activated-potentiated form of antibodies to the brain-specific protein S-100 according to the invention is made in the form of a pharmaceutical composition and contains an activated-potentiated form of antibodies to endothelial NO synthase as an additional reinforcing component.

In addition, the activated-potentiated form of antibodies to endothelial NO synthase and the activated-potentiated form of antibodies to brain-specific protein S-100 are used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution, the activity of which is due to the process of multiple sequential dilutions in aqueous or aqueous alcohol solvent in combination with external mechanical action - vertical shaking of each dilution.

Moreover, the pharmaceutical composition can be made in solid dosage form and contains an effective amount of granules of a neutral carrier saturated with a mixture of aqueous or aqueous-alcoholic solutions of an activated-potentiated form of antibodies to endothelial NO synthase and an activated-potentiated form of antibodies to brain-specific protein S-100, and pharmaceutically acceptable additives including lactose, microcrystalline cellulose and magnesium stearate.

Moreover, aqueous or aqueous-alcoholic solutions of activated-potentiated forms of antibodies to endothelial NO synthase and brain-specific protein S-100 are obtained by repeated serial dilutions and external influences — vertical shaking of each dilution from matrix solutions of affinity-purified antibodies to endothelial NO synthase and to brain-specific protein S-100 with a concentration of 0.5 ÷ 5.0 mg / ml

Each of the components of the claimed neurotropic drug is used in the form of a mixture of various, mainly hundred, dilutions according to homeopathic technology.

In addition, a pharmaceutical composition comprising an activated-potentiated form of antibodies to the brain-specific protein S-100 and an activated-potentiated form of antibodies to endothelial NO synthase is used to treat organic diseases of the nervous system.

Moreover, the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used, preferably, for the treatment of Parkinson's disease.

Moreover, the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat the psycho-organic syndrome.

Moreover, a pharmaceutical composition containing an activated-potentiated form of antibodies to the brain-specific protein S-100 and an activated-potentiated form of antibodies to endothelial NO synthase is used to treat cerebrovascular diseases, preferably for the treatment of acute cerebrovascular accident - stroke.

In addition, a pharmaceutical composition comprising an activated-potentiated form of antibodies to the brain-specific protein S-100 and an activated-potentiated form of antibodies to endothelial NO synthase is used to treat encephalopathies of various origins.

The solution to this problem is also ensured by the fact that in the method of treating organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various origins by introducing into the body a neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 according to the invention, an activated -potentiated form of antibodies to endothelial NO synthase.

In this case, the activated-potentiated form of antibodies to endothelial NO synthase and the activated-potentiated form of antibodies to brain-specific protein S-100 are used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution of each component, the activity of which is due to the process of repeated dilution in aqueous or aqueous alcohol solvent in combination with external mechanical action - vertical shaking of each dilution.

In addition, they use a mixture of different dilutions of antibodies to endothelial NO synthase prepared in the form of a single drug - a single dosage form in combination with a mixture of different dilutions of antibodies to the brain-specific protein S-100, prepared according to homeopathic technology.

Moreover, a neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used, preferably, for the treatment of Parkinson's disease.

In addition, a neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat cerebrovascular diseases, preferably for the treatment of acute cerebrovascular accident - stroke.

The drug may contain active ingredients in a volume ratio of 1: 1, with each component used in the form of a mixture of three matrix solutions, diluted, respectively, 100 ¹² , 100 ³⁰ and 100 ²⁰⁰ times, which is equivalent to hundreds of dilutions of C12, C30, C200, cooked by homeopathic technology.

The claimed pharmaceutical composition is recommended to be taken, preferably, 1-2 tablets 2-4 times a day.

In the treatment of organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various origins, separate but combined and simultaneous use (administration to the body) of the claimed pharmaceutical composition in the form of two separately prepared preparations, both in the form of solutions, and in solid dosage forms (tablets), each of which contains an activated-potentiated form of ultra-low doses of affinity-purified antibodies to the brain-specific protein S-100 and, accordingly, an activated-potentiator form of ultra-low doses of affinity-purified antibodies to endothelial NO synthase.

The proposed combination of activated-potentiated forms of antibodies to the brain-specific protein S-100 and to endothelial NO synthase in a pharmaceutical composition (i.e., forms of antibodies to the brain-specific protein S-100 and to endothelial NO synthase prepared according to the homeopathic potentiation technique by repeated dilution and external exposure - repeated vertical shaking of each dilution (see, for example, V. Schwabe. "Homeopathic medicines", M., 1967, p. 14-29), which are active, due to using potentiating technology, in pharmacological models and / or clinical methods of treating psycho-organic syndrome and encephalopathies of various genesis) provides an unexpected synergistic therapeutic effect, confirmed by adequate (valid) experimental models and clinical studies, which consists in increasing the effectiveness of the treatment of organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various origins.

The indicated technical result is provided by an increase in the neuroprotective activity of antibodies to the S-100 protein, due to the influence on the efficiency of the interaction of ligands with the sigma-1 receptor, an increase in the vegetative stabilizing effect, normalization of the vegetative status, a synergistic effect of both components on neuronal plasticity and, consequently, an increase in brain resistance to toxic effects, which improves integrative activity and restores interhemispheric connections of the brain, helps to injury to cognitive impairment, stimulates reparative processes and accelerates the restoration of central nervous system (CNS) functions, increases mental performance, restores learning and memory processes, normalizes somatovegetative manifestations, increases cerebral blood flow and, accordingly, provides an extension of the therapeutic range of the drug and an increase in the effectiveness of treatment of organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various origins, due to ennyh acute and chronic cerebral circulatory disorders, traumatic brain injuries, neuroinfections, atherosclerotic and neurodegenerative brain injury, cerebrovascular (including ischemic) diseases. The claimed drug can be used in monotherapy of somatoform dysfunction of the autonomic nervous system. In addition, the use of the claimed drug ensures the restoration of integrative activity of the brain with a wide range of organic disorders of the central nervous system. Including the claimed drug can be used as part of complex therapy.

In this case, the claimed drug, as well as its constituent components, does not have a sedative and muscle relaxant effect, does not cause addiction and addiction.

In addition, the claimed drug expands the arsenal of drugs intended for the treatment of organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various origins.

The drug is prepared mainly as follows.

For the preparation of the activated-potentiated form of the active components, monoclonal or, mainly, polyclonal antibodies are used, which can be obtained by known technologies - the techniques described, for example, in the book: Immunological methods, ed. G. Fremel, Moscow: “Medicine”, 1987, p. 9-33; or, for example, in an article by Laffly E., Sodoyer R. Hum. Antibodies. Monoclonal and recombinant antibodies, 30 years after. - 2005 .-- Vol.14. - N 1-2. P.33-55.

Monoclonal antibodies are obtained, for example, using hybridoma technology. Moreover, the initial stage of the process includes immunization, based on the principles already developed in the preparation of polyclonal antisera. Further stages of the work include obtaining hybrid cells producing clones of antibodies of the same specificity. Their isolation in an individual form is carried out by the same methods as in the case of polyclonal antisera.

Polyclonal antibodies can be obtained by active immunization of animals. For this purpose, according to a specially developed scheme, animals are given a series of injections with the antigens required by the invention: brain-specific protein S-100 and endothelial NO synthase. As a result of this procedure, monospecific antisera with a high content of antibodies are obtained, which are used to obtain activated-potentiated forms of components. If necessary, the antibodies present in the antiserum are purified, for example, by affinity chromatography, using fractionation by salt precipitation or ion exchange chromatography.

Preferred for the preparation of the claimed pharmaceutical composition is the use of polyclonal antibodies to the brain-specific protein S-100 and to endothelial NO synthase, which are used as a matrix (primary) solution with a concentration of 0.5 ÷ 5.0 mg / ml for subsequent preparation of activated-potentiated forms.

Preferred for the preparation of each component is the use of a mixture of three water-alcohol dilutions of the initial matrix solution of antibodies, diluted, respectively, 100 ¹² , 100 ³⁰ and 100 ²⁰⁰ times, which corresponds to hundreds of dilutions C12, C30 and C200 prepared according to homeopathic technology. When performing the claimed drug in solid dosage form, a mixture of these components in a volume ratio of 1: 1 can be applied to a neutral carrier - lactose.

Preferred for the preparation of the claimed drug is the use of polyclonal antibodies to endothelial NO synthase and to the brain-specific protein S-100, which can be obtained by immunization of rabbits as follows.

For experimental studies, antibodies were used, prepared by order of a specialized biotechnological company.

Polyclonal antibodies to endothelial NO synthase are prepared using the adjuvant and the whole endothelial NO synthase molecule of the following sequence as immunogen (antigen) for rabbit immunization:

It is possible to obtain polyclonal antibodies to endothelial NO synthase using the following sequence as an immunogen (antigen) of a whole molecule of endothelial NO synthase:

It is possible to obtain polyclonal antibodies to endothelial NO synthase using, as an immunogen (antigen), a synthetic peptide of endothelial NO synthase, selected, for example, from the following amino acid sequences:

Before blood sampling for 1-3 days, 1-3 intravenous injections are performed to increase the level of antibodies. During immunization, small blood samples are taken from rabbits to evaluate the amount of antibody. The maximum level of the immune response to the administration of most soluble antigens is achieved 40-60 days after the first injection. After the end of the first cycle of immunization of rabbits for 30 days, they restore health and re-immunization, including 1-3 intravenous injections. To obtain antisera from immunized rabbits, blood is collected in a 50 ml centrifuge tube. Using a wooden spatula, the formed clots are removed from the walls of the tube and the wand is placed in the clot formed in the center of the tube. Blood is placed in a refrigerator (temperature 4 ° C) at night. The next day, the clot attached to the spatula is removed and the remaining liquid is centrifuged at 13000g for 10 minutes. The supernatant (supernatant) is an antiserum. The resulting antiserum should be yellow. Add to the antiserum 20% (weight concentration) NaN ₃ to a final concentration of 0.02% and store until use in a frozen state at a temperature of -20 ° C (or without adding NaN ₃ at a temperature of -70 ° C). To isolate antibodies to endothelial NO synthase from antiserum, solid phase absorption is performed in the following sequence:

1) 10 ml of rabbit antiserum is diluted 2 times with 0.15 M NaCl, 6.26 g of Na ₂ SO ₄ are added, stirred and incubated for 12-16 hours at 4 ° C;

2) the precipitate formed is removed by centrifugation, dissolved in 10 ml of phosphate buffer and then dialyzed against the same buffer overnight at room temperature;

3) after removing the precipitate by centrifugation, the solution is applied to a column with DEAE-cellulose, balanced with phosphate buffer;

4) the antibody fraction is determined by measuring the optical density of the eluate at 280 nm.

The antibodies are then purified by affinity chromatography by attaching the obtained antibodies to endothelial NO synthase, which is located on an insoluble matrix, followed by elution with concentrated salt solutions.

Thus obtained buffer solution of polyclonal rabbit antibodies to endothelial NO synthase purified on an antigen with a concentration of 0.5 ÷ 5.0 mg / ml, preferably 2.0 ÷ 3.0 mg / ml, is used as a matrix (primary) solution for the subsequent preparation of the activated-potentiated form according to homeopathic technology.

To obtain polyclonal antibodies to the brain-specific protein S-100, the brain-specific protein S-100 is used, the physicochemical properties of which are described in the article by M.V. Starostina, S.M. Sviridov, Neurospecific protein S-100, Advances in modern biology, 1977, v.5, p.170-178; book of MB Stark, Brain-specific proteins / antigens / and neuron functions, M .: "Medicine", 1985, p.12-14, isolated from bull brain tissue by the following method:

- fabric frozen in liquid nitrogen is ground into powder in a special mill;

- the extraction of proteins is carried out in a ratio of 1: 3 (weight / volume) in the extraction buffer during homogenization;

- the homogenate is heated for 10 min at 60 ° C and cooled to 4 ° C in an ice bath;

- thermolabile proteins are removed by centrifugation;

- carry out stepwise fractionation with ammonium sulfate, followed by removal of precipitated impurity proteins;

- the S-100 protein-containing fraction is precipitated with 100% saturated ammonium sulfate when the pH is reduced to 4.0 and collected by centrifugation;

- the precipitate is dissolved in a minimum volume of a buffer containing EDTA and mercaptoethanol, dialyzed against deionized water and freeze-dried;

- fractionation of acidic proteins is continued by chromatography on ion-exchange media — DEAE cellulose DE-52 and then DEAE-Sephadex A-50;

- collected and dialyzed fractions containing S-100 protein are separated by molecular weight by gel filtration on Sephadex G-100;

- purified S-100 protein is dialyzed and freeze-dried.

The molecular weight of the purified brain-specific protein S-100 is 21,000 Da.

Due to the high content of aspartic and glutamic acids, the brain-specific protein S-100 is strongly acidic and occupies the extreme anode position during electrophoresis in a discontinuous buffer system in a polyacrylamide gel, which facilitates its identification.

To obtain a brain-specific antiserum, a purified S-100 protein (antigen) is prepared in combination with methylated bovine serum albumin as a carrier with Freund's complete adjuvant, which is administered subcutaneously to a laboratory rabbit animal in the amount of 1- to a selected brain-specific protein S-100. 2 ml On the 8th, 15th day, re-immunization is carried out. Blood is taken (for example, from an ear vein) on the 26th and 28th day.

The resulting antiserum has a titer of 1: 500 - 1: 1000, forms a single precipitation band with extract from the nervous tissue, but does not react with extracts of heterologous organs, and forms a single precipitation peak with both pure S-100 protein and the extract of nervous tissue, which indicates the monospecificity of the obtained antiserum.

The activated-potentiated form of antibodies of each component (i.e., the forms of antibodies to bradykinin, to morphine, and to histamine prepared using the homeopathic potentiation technique by repeated serial dilution in combination with external exposure - repeated vertical shaking of each component (see, for example, B . Schwabe. "Homeopathic medicines", M., 1967, p.14-29), which have activity due to potentiation technology in pharmacological models and / or clinical methods of diseases of the respiratory tract) is prepared by uniformly decreasing the concentration as a result of successive dilutions of 1 part of the above matrix solution in 9 parts (for decimal dilution D) or 99 parts (for hundredth dilution C) or 999 parts (for thousandth dilution M) of a neutral solvent in combination with repeated (at least 10 times) vertical shaking ("dynamization") of each dilution obtained and the use of separate containers for each subsequent dilution to obtain the desired otentsii - dilution multiplicity of homeopathic method (see, e.g., W. Schwabe.. "Homeopathic medicines", M., 1967, p.14-29).

External processing in the process of reducing the concentration can also be performed by ultrasound, electromagnetic or other physical effects.

For example, to prepare the 12th hundredth dilution of C12, one part of the above matrix solution of antibodies to NO synthase (or to brain-specific protein S-100) with a concentration of 2.5 mg / ml is diluted in 99 parts of a neutral aqueous or aqueous-alcoholic solvent (mainly 70% ethyl alcohol) and repeatedly (10 or more times) vertically shake - potentiate the obtained 1st hundredth C1 dilution. From the 1st hundredth C1 dilution prepare the 2nd hundredth dilution C2. This operation is repeated 11 times, obtaining the 12th hundredth dilution of C12. Thus, the 12th hundredth dilution of C12 is a solution obtained by diluting successively in different containers 12 times of the first part of the initial matrix solution of antibodies to NO synthase with a concentration of 2.5 mg / ml in 99 parts of a neutral solvent, t .e. a solution prepared by diluting the matrix solution in 100 ¹² times. Similar operations with the appropriate dilution ratio are carried out to obtain dilutions of C30 and C200.

When using as a component (for example, an activated-potentiated form of antibodies to the brain-specific protein S-100) mixtures of different, mainly hundred, dilutions, each dilution of the component (for example, C12, C30, C200) is prepared separately according to the technology described above to their penultimate dilution (respectively, to obtain C11, C29, C199) and then add one part of each penultimate dilution obtained in one container and mix with the required amount of solvent (respectively, with 97 parts for hundreds of dilution). In this case, an activated-potentiated form of antibodies to the brain-specific protein S-100 is obtained in an ultra-low dose obtained by super-dilution of the matrix solution, respectively, 100 ¹² , 100 ³⁰ and 100 ²⁰⁰ times, equivalent to a mixture of hundred-percent dilutions of C12, C30 and C200, using homeopathic technology.

It is possible to use each component in the form of a mixture of other different dilutions according to homeopathic technology, for example decimal and or hundredths (C12, C30, C100; D20, C30, C100 or D20, C30, M100, etc.), the effectiveness of which is determined experimentally.

To obtain the claimed pharmaceutical composition, aqueous or aqueous-alcoholic solutions of the active ingredients are mixed, mainly in a volume ratio of 1: 1 and used in liquid dosage form.

The claimed pharmaceutical composition can be used in solid dosage form, which contains an effective amount of particles of a neutral carrier - lactose, saturated by impregnation of an activated-potentiated form of antibodies to endothelial NO synthase with an activated-potentiated form of antibodies to a mixture of aqueous or aqueous-alcoholic solutions brain-specific protein S-100, and pharmaceutically acceptable additives, including, mainly, lactose, microcrystalline cellulose and magnesium with earat.

To obtain a solid oral form of the claimed medicinal product, a fluidized bed is installed (for example, Huttlin Pilotlab type manufactured by Huttlin GmbH) irrigation until saturation of particles of a neutral substance - lactose (milk sugar) with a particle size of 100 ÷ 300 is introduced into the fluidized - boiling layer μm, a pre-prepared aqueous or aqueous-alcoholic solution of activated-potentiated forms of antibodies to the brain-specific protein S-100 and to endothelial synthase of nitric oxide (NO synthase), mainly in the ratio 1 kg of antibody solution per 5 or 10 kg of lactose (1: 5 - 1:10) with simultaneous drying in a stream of heated air supplied under the grate at a temperature not exceeding 40 ° C. The estimated amount of 10-34 wt. parts of dried granules saturated with the activated-potentiated form of antibodies are loaded into the mixer and mixed with 25 ÷ 85 wt. parts of the total load mass of “unsaturated” pure lactose (to reduce the cost and some simplification and acceleration of the process without reducing the effectiveness of the therapeutic effect). Then, magnesium stearate is added to this mixture in an amount of 0.1 ÷ 1 wt. parts of the total mass of the load and microcrystalline cellulose in an amount of 3 ÷ 10 wt. parts of the total mass of the load. The resulting tablet mass is uniformly mixed and tabletted by direct dry pressing (for example, in a Korsch tablet press - XL 400) with the formation of round tablets weighing 150 ÷ 500 mg, mainly weighing 300 mg, soaked in an aqueous-alcoholic solution (3.0-6.0 mg / table.) of the activated-potentiated form of antibodies to the brain-specific protein S-100 and to NO synthase in an ultra-low dose, each component prepared from a matrix solution diluted, respectively, in 100 ¹² , in 100 ³⁰ and in 100 ²⁰⁰ , which equivalent to a mixture of hundred dilutions C12, C30 and C200, pr tovlennyh by homeopathic technology.

Glucose, sucrose, maltose, starch, isomaltose, isomalt and other monosaccharides, oligosaccharides and polysaccharides used in the manufacture of pharmaceutical preparations, as well as technological mixtures based on the latter with the addition of pharmaceutically acceptable additives (for example, isomalt, crospovidone, can also be used as a neutral carrier. , sodium cyclamate, sodium saccharin, anhydrous citric acid, etc.), including lubricants, disintegrants, binders and dyes.

Preferably, the claimed pharmaceutical composition is recommended to take 1-2 tablets 2-4 times a day.

For experimental studies, antibodies were used, prepared by order of a specialized biotechnological company.

Example 1

Investigation of the effect of a complex preparation, which includes activated-potentiated forms of polyclonal affinity purified rabbit antibodies to brain-specific protein S-100 (anti-S100) and to endothelial NO synthase (anti-eNOS) in ultra-low doses (SMD) obtained by super-dilution of the original matrix solution (concentration of 2.5 mg / ml) in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of hundreds of homeopathic dilutions C12, C30, C200, in a ratio of 1: 1 (SMD anti-S100 + anti-eNOS), and also the components that make up its activated-potenti bath forms of polyclonal rabbit antibodies to brain-specific protein S-100, purified on antigen, in an ultra-low dose (SMD anti-S100) obtained by super-dilution of the initial matrix solution 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of hundred-percent homeopathic dilutions C12, C30 , C200 and an activated-potentiated form of polyclonal rabbit antibodies to endothelial NO synthase purified on an antigen in an ultra-low dose (SMD anti-eNOS) obtained by super-dilution of the initial matrix solution 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of cell All homeopathic dilutions of C12, C30, C200 were performed in vitro to bind the standard [ ³ H] pentazocine ligand to the recombinant human sigma-1 receptor, and evaluated by the radioligand method.

Sigma-1 receptor is an intracellular receptor localized in cells of the central nervous system, cells of most peripheral tissues and immunocompetent cells. Receptors demonstrate a unique ability to translocate, which is caused by many psychotropic drugs. The dynamics of sigma-1 receptors is directly related to the various effects carried out by the drugs when they act on sigma-1 receptors. These effects include regulation of activity channels, ecocytosis, signaling, plasma membrane remodeling (raft formation), and lipid transport / metabolism. All this can contribute to the development of plasticity of neurons in the brain. There is evidence that sigma-1 receptors have a modulating effect on all major neurotransmitter systems: noradrenergic, serotonergic, dopaminergic, cholinergic systems and NMDA-regulated glutamate effects. Sigma-1 receptor plays an important role in the pathophysiology of neurodegenerative diseases, in particular Parkinson's disease, mental and affective disorders, stroke, is involved in learning and memory. In this regard, the ability of drugs to influence the effectiveness of the interaction of ligands with the sigma-1 receptor indicates the presence of neuroprotective, anti-ischemic, anxiolytic, antidepressant, anti-asthenic components in the spectrum of their pharmacological activity, which allows us to consider these drugs as effective drugs, including including for the treatment of cerebrovascular disease.

Potentiated distilled water (a mixture of homeopathic dilutions C12 + C30 + C200) was tested as a control of the tested drugs.

In the experiment (for measuring total binding), 20 μl of the complex preparation of SMD anti-S100 + anti-eNOS or 10 μl of SMD AT to S100 or 10 μl of SMD AT to NOS was added to the incubation medium. Thus, the amount of SMD anti-S100 + anti-eNOS introduced into the experimental well when testing the complex preparation was identical to the number of SMD AT to S100 and SMD AT to NOS tested as single drugs, which allows us to compare the effectiveness of the complex preparation with its individual the components that make up its composition. Potentiated distilled water was introduced into the incubation medium in a volume of 20 μl and 10 μl.

Then 160 μl (~ 200 μg protein) of the homogenate of the cell membranes of the Jurkat line (human leukemic T-lymphocyte line), and lastly 20 μl of the tritium-labeled radioligand, [ ³ H] pentazocine (15 nM) were added.

To measure nonspecific binding, instead of preparations or potentized water, 20 μl of unlabeled ligand, haloperidol (10 μM), was added to the incubation medium.

Radioactivity was measured on a scintillation counter (Topcount, Packard) using a scintillation mixture (Microscint 0, Packard) after incubation for 120 minutes at 22 ° C in 50 mM Tris-HCl buffer (pH 7.4) and filtration on glass fiber filters ( GF / B, Packard). Specific binding (in experiment or control) was calculated as the difference between total (in experiment or control) and non-specific binding.

The results are presented as percent inhibition of specific binding in the control (distilled water was used as a control) (Table 1).

Table 1 The effect of drugs and potentiated water on the binding of a standard radioligand [ ³ H] pentazocine to a recombinant human sigma-1 receptor Experimental group The amount introduced into the experimental well % of specific radioligand binding in the control % inhibition of radioligand binding in the control 1st dimension 2nd dimension Average value SMD anti-S100 + anti-eNOS 20 μl 48,4 35.5 42.0 58.0 SMD anti-S100 10 μl 67.3 63.1 65,2 34.8 SMD anti-eNOS 10 μl 147.5 161.1 154.3 -54.3 Potentized water 20 μl 98.1 75.8 86.9 13.1 Potentized water 10 μl 140.1 106,2 123,2 -23.2 Note: % specific binding in the control = (specific binding in the experiment / specific binding in the control) * 100%; % inhibition of specific binding in the control = 100% - (specific binding in the experiment / specific binding in the control) * 100%).

Results reflecting inhibitions above 50% represent significant effects of the test compounds; inhibition from 25% to 50%, indicates the effects of mild to moderate; inhibition of less than 25% is considered a negligible effect of the test compound and is within the background level.

Thus, under the conditions of this experimental model, it was shown that the complex preparation of SMD anti-S100 + anti-eNOS is more effective than its individual components (SMD anti-S100 and SMD anti-eNOS) and inhibits the binding of the standard radioligand [ ³ H] pentazocine to human recombinant sigma-1 receptor; SMD anti-S100, introduced into the experimental well in a volume of 10 μl, inhibits the binding of the standard radioligand [ ³ H] pentazocine to the recombinant human sigma-1 receptor, but the severity of the effect is less than that of the complex drug SMD anti-S100 + anti-eNOS; SMD anti-eNOS introduced into the experimental well in a volume of 10 μl did not affect the binding of the standard radioligand [ ³ H] pentazocine to the recombinant human sigma-1 receptor; potentiated water introduced into the experimental well in a volume of 10 μl or 20 μl did not affect the binding of the standard radioligand [ ³ H] pentazocine to the recombinant human sigma-1 receptor.

Example 2

To study the properties of the claimed drug in the treatment of acute cerebrovascular accident - stroke, 300 mg tablets were used, impregnated with a pharmaceutical composition containing water-alcohol solutions (6 mg / tablet) of activated-potentiated forms of polyclonal affinity-purified rabbit antibodies to brain-specific protein S -100 (anti-S100) and to endothelial NO synthase (anti-eNOS) in ultra-low doses (SMD) obtained by super-dilution of the initial matrix solution (with a concentration of 2.5 mg / ml) in 100 ^{1 2} , 100, ³⁰ , 100, ²⁰⁰ times, equivalent to a mixture of hundreds of homeopathic dilutions C12, C30, C200 (SMD anti-S100 + anti-eNOS).

The study included patients with a diagnosis of acute cerebrovascular accident (stroke) according to the ischemic type in the system of the right internal carotid artery (ischemic stroke).

As a control group, we used a group of patients receiving standardized vascular-metabolic therapy, including non-narcotic analgesics and non-steroidal anti-inflammatory drugs, antiplatelet agents, anticoagulants and neuroprotective drugs.

By design, the study was an open, randomized comparative clinical study evaluating the effectiveness of the use of the drug SMD anti-S100 + anti-eNOS in combination with complex vascular and metabolic therapy compared to the complex vascular and metabolic therapy itself (SMT) in the treatment of patients in the early recovery period ischemic stroke

The study included 12 patients with a diagnosis of acute cerebrovascular accident according to ischemic type in the system of the right internal carotid artery aged 50 to 84 years. The average age of the patients was 66.66 ± 9.3 years.

The study examined patients' compliance with the following inclusion and exclusion criteria.

Inclusion criteria

1. Patients with acute cerebrovascular accident according to ischemic type in the system of the right internal carotid artery, 21 days old by the time of visit 1, confirmed by medical history, neurological examination and medical documentation.

2. Patients from the first day of the disease receive complex vascular-metabolic therapy in the framework of the standards of medical care in Moscow.

3. Right-handed patients, with confirmation by a study of the lateralization of sensorimotor functions according to T. A. Dobrokhotova and N. N. Bragina.

4. The etiological factor of stroke in a patient is a combination of atherosclerosis and arterial hypertension.

5. Patients with left-sided hemiparesis with increased muscle tone in spastic type at visit 1.

6. Integral assessment according to the Lindmark scale at visit 0 is not lower than 345 points.

7. Integral assessment on the Lindmark scale at visit 1 from 345 to 404 points.

8. The lack of prerequisites for the appointment of immunomodulating agents over the next 6 months.

9. The level of education and the degree of understanding of the patient are sufficient to adequately communicate with the researcher and the coordinator of the study.

10. Patients are evaluated by the researcher as reliable, ready to perform all prescribed clinical visits, tests and procedures provided for by the protocol.

11. Patients have a valid home address.

12. Signed informed consent form.

Exclusion criteria

1. A history of surgical intervention on the brain.

2. Acute myocardial infarction.

3. Hemorrhagic heart attack.

4. A history of psychosis, bipolar disorder, or schizoaffective disorder.

5. Major depressive disorder according to the criteria for the depression module of the international neuropsychiatric mini-interview (MINI).

6. Factors / conditions of a medical or other nature that, in the opinion of the researcher, may affect the patient's test results in this study.

7. Answers “2A”, “2B”, “2B” or “3” in the heading “I” of the Beck Depression Questionnaire (active suicidal thinking with some intention to act, without a specific plan or active suicidal thinking with a specific plan and intent).

8. A history of autoimmune disease.

9. Acute liver damage or severe cirrhosis (Child-Pugh class C).

10. Unregulated thyroid dysfunction.

11. History of uncompensated arterial hypertension.

12. Serious or decompensated cardiovascular disease, liver, kidney disease, metabolic, respiratory or hematological disease, symptomatic peripheral vascular disease, or other medical or psychiatric condition that, in the opinion of the researcher, may affect the patient’s participation in the study or may lead to prolongation of hospitalization or re-hospitalization of the patient during the study.

13. Diseases and conditions that, in the opinion of the researcher, may interfere with the participation of the patient in the study.

14. Taking Impaz (SMD anti-eNOS) or Tenoten (SMD anti-S100) before taking the study.

15. Admission of antidepressants of any group, not excluding herbal and homeopathic medicines.

16. Reception of anxiolytics of any group, not excluding herbal and homeopathic preparations.

17. Reception of immunomodulators, not excluding herbal and homeopathic preparations.

18. Systemic treatment with steroids for 1 month or less before visit 0.

19. Vaccinations, including against the flu.

20. Participation in the study of Impaz (SMD anti-eNOS) or Tenoten (SMD anti-S100), if patients have taken at least one dose of the drug.

21. Participation in other clinical trials for 1 month before inclusion in this study.

22. Pregnancy, breast-feeding, the inability to use adequate contraception during the study and within a month after the last administration of the study drug.

23. The presence of allergies / intolerances to any of the components of the drugs used in the treatment, including lactose intolerance.

24. The use of drugs, antipsychotics, alcoholism, mental illness of the patient.

25. Patients are the staff of the center directly related to the study, and / or are members of the family of the staff of the research center directly related to the study. The concept of "family members" includes a spouse (s), parents, children, brothers (sisters) associated with both biological and legal kinship.

26. Participation in litigation or alleged compensation or participation in litigation, according to the researcher.

After determining whether the patient met the criteria for inclusion and exclusion of the protocol, the patients were randomized into two study groups: a group of patients receiving SMD anti-S100 + anti-eNOS in combination with standardized vascular and metabolic therapy (6 people, women - 33.33%, men - 66.66%, average age - 65.33 ± 6.71 years), a group of patients receiving actually standardized vascular-metabolic therapy (SMT) (6 people, women - 50%, men - 50%, average age - 68 , 0 ± 11.88 years).

In the course of this study, 5 visits were made to patients. The treatment phase lasted from Visit 1 to Visit 4 for 84 ± 5 days. Visit 4 (day 84 ± 5) was the first endpoint of the study, after which the follow-up phase began. The follow-up phase continued until Visit 5 (day 168 ± 5).

The safety analysis included data from all patients participating in the study (n = 12). During the entire period of observation of patients, good tolerability of the drug was noted. Adverse events were absent. All patients of the study groups completed the treatment within the time period established by the study protocol; there were no prematurely retired patients.

When assessing the effect of the drug SMD anti-S100 + anti-eNOS on the main manifestations of stroke (Lindmark scale), anxiety-depressive disorders of the patient (Beck depression questionnaire), as well as the patient's cognitive functions (MMSE questionnaire), an improvement was found in the main manifestations of stroke, expressed in a statistically significant decrease in the overall Lindmark score (from 383.5 ± 7.81 to 412.33 ± 7.47, p <0.001), the severity of anxiety-depressive disorders, expressed in a statistically significant decrease in the overall score of the Beck depression questionnaire (s 12.16 ± 3.6 to 6.33 ± 4.76, p <0.05), and also a statistically significant improvement in the cognitive sphere, expressed in an increase in the total score for the MMSE questionnaire (from 26.0 ± 3.69 to 29.66 ± 0.52, p <0.05) by the end of treatment (Table 2).

Moreover, in the control group only a statistically significant trend of influence on the main manifestations of ONMK (Lindmark scale) was shown in the form of an increase in the total score (from 379.5 ± 8.12 to 394.66 = 13.67, p <0.05).

An intergroup comparison showed a significant difference in the dynamics of the main manifestations of stroke in the Lindmark scale (compared with the control p <0.02). The differences between the groups in the dynamics of anxiety-depressive disorders did not reach statistically significant values, which, apparently, is associated with an insufficient number of patients.

The described changes did not undergo significant differences during the follow-up period.

table 2 Lindmark scale Beck Depression Questionnaire MMSE SMD anti-S100 + anti-eNOS before treatment 383.5 ± 7.81 12.16 ± 3.6 26.0 ± 3.69 SMD anti-S100 + anti-eNOS after treatment 412.33 ± 7.47 ** # 6.33 ± 4.76 * 29.66 ± 0.52 * SMT before treatment 379.5 ± 8.12 11.66 ± 3.67 27.83 ± 2.48 SMT after treatment 394.66 = 13.67 * 10.16 ± 2.714 28.5 ± 2.07 * - p from ref. <0.05 ** - p from ref. 0.001 # - p compared to control <0.02

Thus, the clinical study showed the positive effect of the combined preparation of SMD anti-S100 + anti-eNOS in combination with standardized vascular metabolic therapy (SMT) on the main manifestations of stroke and the possible effect on anxiety-depressive disorders and cognitive functions of patients in early recovery period of ischemic stroke. In addition, high tolerance to the drug has been confirmed. Adverse events when taking the drug SMD anti-S100 + anti-eNOS were absent.

Example 3

To study the properties of the claimed drug in the treatment of Parkinson's disease, 300 mg tablets were used, impregnated with a pharmaceutical composition containing water-alcohol solutions (6 mg / tablet) of activated-potentiated forms of polyclonal affinity-purified rabbit antibodies to brain-specific protein S-100 (anti -S100) and to endothelial NO-synthase (anti-eNOS) in ultra low doses (RMA) obtained sverhrazvedeniem initial matrix solution (2.5 mg / ml) 100 ^12, 100 ^30, 100 ^{to 200} times equivalent mixtures centesimal homeopathic dilutions C12, C30, C200 (SMD anti-S100 + anti-eNOS).

As a control group, we used a group of patients receiving the drug in the form of tablets soaked in an aqueous-alcoholic solution (6 mg / tablet) of an activated-potentiated form of polyclonal rabbit antibodies to brain-specific protein S-100, purified on antigen, in an ultra-low dose (SMD anti -S100) obtained by super-dilution of the initial matrix solution 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of hundred homeopathic dilutions C12, C30, C200.

Patients with an established diagnosis of Parkinson's disease were included in the study. Parkinson's disease is characterized by progressive destruction and death of dopamine neurons in the central nervous system.

By design, the study was a single-center, open, randomized comparative clinical study of the efficacy and safety of the use of anti-S100 + anti-eNOS SMD and anti-S100 SMD drugs used as monotherapy, together with dopamine receptor agonists or together with levodopa drugs in the treatment of patients with Parkinson's disease.

The study included 14 patients with a diagnosis of Parkinson's disease, aged 52 to 80 years. The average age of the patients was 67.93 ± 11.56 years.

The study examined patients' compliance with the following inclusion and exclusion criteria.

Inclusion criteria

1. Patients with Parkinson's disease, stages 1 to 5 according to Hen-Yar, confirmed by the medical history (including the test with levodopa), neurological examination and medical documentation.

2. The form of Parkinson's disease is predominantly tremulous, trembling-rigid or akinetic-rigid.

3. Hyun-Yar stages for patients not receiving antiparkinsonian therapy from 1 to 2.5 inclusive at visit 1.

4. Hyun-Yar stages for patients receiving antiparkinsonian therapy from 1 to 5 inclusive at visit 1.

5. Lack of prerequisites for the appointment of immunomodulating agents over the next 6 months.

6. The level of education and the degree of understanding of the patient are sufficient to adequately communicate with the researcher and the coordinator of the study.

7. Patients are evaluated by the researcher as reliable, ready to perform all the prescribed clinical visits, tests and procedures prescribed by the protocol.

8. Patients have a valid home address.

9. Signed informed consent form.

Exclusion criteria

1. A history of surgical intervention on the brain.

2. A history of stroke.

3. A history of psychosis, bipolar disorder, or schizoaffective disorder.

4. Major depressive disorder according to the criteria for the depression module of the international neuropsychiatric mini-interview (MINI).

5. Factors / conditions of a medical or other nature that, in the opinion of the researcher, may affect the patient's test results in this study.

6. Answers “2A”, “2B”, “2B” or “3” in the heading “I” of the Beck Depression Questionnaire (active suicidal thinking with some intention to act, without a specific plan or active suicidal thinking with a specific plan and intent).

7. A history of autoimmune disease.

8. Acute liver damage or severe cirrhosis (Child-Pugh class C).

9. Unregulated thyroid dysfunction.

10. An uncompensated history of hypertension.

11. Serious or decompensated cardiovascular disease, liver, kidney disease, metabolic, respiratory or hematological disease, symptomatic peripheral vascular disease, or other medical or psychiatric condition that, in the opinion of the researcher, may affect the patient’s participation in the study or may lead to prolongation of hospitalization or re-hospitalization of the patient during the study.

12. Diseases and conditions that, according to the researcher, may interfere with the participation of the patient in the study.

13. Taking Impaz (SMD anti-eNOS) or Tenoten (SMD anti-S100) before taking the study.

14. Admission of antidepressants of any group, not excluding herbal and homeopathic medicines.

15. Reception of anxiolytics of any group, not excluding herbal and homeopathic preparations.

16. Reception of immunomodulators, not excluding herbal and homeopathic preparations.

17. Systemic steroid treatment for 1 month or less prior to visit 1.

18. Vaccinations, including against the flu.

19. Participation in the study of Impaz (SMD anti-eNOS) or Tenoten (SMD anti-S100), if patients have taken at least one dose of the drug.

20. Participation in other clinical trials for 1 month before inclusion in this study.

21. Pregnancy, breast-feeding, the inability to use adequate contraception during the study and within a month after the last administration of the study drug.

22. The presence of allergies / intolerances to any of the components of the drugs used in the treatment, including lactose intolerance.

23. The use of drugs, antipsychotics, alcoholism, mental illness of the patient.

24. Patients are the staff of the center directly related to the study, and / or are members of the family of the staff of the research center directly related to the study. The concept of "family members" includes a spouse (s), parents, children, brothers (sisters) associated with both biological and legal kinship.

25. Participation in litigation or alleged compensation or participation in litigation, according to the researcher.

After determining whether the patient met the criteria for inclusion and exclusion of the protocol, patients were randomized into two study groups: a group of patients who received anti-S100 SMD in combination with anti-Parkinsonian therapy (7 people, 57.14% women, 42.86% men, medium age - 65.25 ± 10.6 years), and a group of patients who received SMD anti-S100 + anti-eNOS in combination with anti-Parkinsonian therapy (7 people, women - 57.14%, men - 42.86%, average age - 71.5 ± 12.79 years).

During this study, 4 visits to the research center were conducted. The treatment phase lasted from Visit 1 to Visit 3 for 56 ± 5 days. Visit 3 (day 56 ± 5) was the first endpoint of the study, after which the follow-up phase began. Follow-up phase continued until Visit 4 (day 84 ± 5).

The safety analysis included data from all patients participating in the study (n = 14). During the entire period of observation of patients, good tolerability of the drug was noted. Adverse events were absent. All patients of the study groups completed the treatment within the time period established by the study protocol; there were no prematurely retired patients.

When planning the study and recruiting patients, the groups were divided into the following subgroups:

1. Patients in the early stages of Parkinson's disease (1-2.5 according to Hen-Yar) who are not receiving anti-Parkinsonian therapy, who will receive the drug SMD anti-S100 + anti-eNOS as monotherapy. In a subgroup of 2 patients.

2. Patients at any stage of Parkinson's disease (1-5 according to Hen-Yar), receiving levodopa drugs, in whom the treatment is not accompanied by complications of antiparkinsonian therapy, to which the drug SMD anti-S100 + anti-eNOS will be added. In a subgroup of 2 patients.

3. Patients at any stage of Parkinson's disease (1-5 according to Hen-Yar), receiving levodopa drugs, with complications of anti-Parkinsonian therapy (dyskinesia, on-off periods, hallucinations, etc.), to whom anti-SMD will be added to therapy -S100 + anti-eNOS. In a subgroup of 2 patients.

4. Patients at any stage of Parkinson's disease (1-5 according to Hen-Yar), receiving dopamine receptor agonists as monotherapy, to which anti-S100 + anti-eNOS SMD will be added to therapy. Subgroup 1 patient.

5. Patients in the early stages of Parkinson's disease (1-2.5 according to Hen-Yar) who are not receiving anti-Parkinsonian therapy, who will receive the drug SMD anti-S100 as monotherapy. Subgroup 1 patient.

6. Patients at any stage of Parkinson's disease (1-5 according to Hen-Yar), receiving levodopa drugs, in whom treatment is not accompanied by complications of antiparkinsonian therapy, to which anti-S100 SMD will be added to therapy. In a subgroup of 2 patients.

7. Patients at any stage of Parkinson's disease (1-5 according to Hen-Yar), receiving levodopa drugs, with complications of anti-Parkinsonian therapy (dyskinesia, on-off periods, hallucinations, etc.), to whom anti-SMD will be added to therapy -S100. In a subgroup of 2 patients.

8. Patients at any stage of Parkinson's disease (1-5 according to Hyun-Yar), receiving dopamine receptor agonists as monotherapy, to which anti-S100 SMD will be added to therapy. In a subgroup of 2 patients.

When evaluating the effect of the drug SMD anti-S100 + anti-eNOS on the main manifestations of Parkinson's disease (UPDRS questionnaire), the activity of the patient's daily life (Schwab-Ingland questionnaire), as well as anxiety-depressive disorders of the patient (Beck depression questionnaire), an improvement in the main manifestations of Parkinson's disease, expressed in a statistically significant decrease in the total score of the UPDRS questionnaire (from 63.5 ± 9.77 to 49.16 ± 10.09, p <0.05), and an increase in the level of daily activity of the patient, expressed in a statistically significant increasing score n the Schwab-Ingland scale (from 65.0 ± 12.25 to 78.33 ± 7.53, p <0.05) to Visit 3. In addition, a tendency to a decrease in the severity of anxiety-depressive disorders was shown, which did not reach statistically significant values (possibly due to an insufficient number of patients and a large scatter of initial values) (Table 3).

Moreover, the difference between the groups according to the total score of the UPDRS questionnaire by the end of therapy was significant with a probability of p <0.05.

Similar indicators in the group of patients receiving anti-S100 SMD did not show improvement trends, except for a statistically significant decrease in the level of anxiety-depressive disorders in the Beck Depression Questionnaire (from 12.0 ± 7.08 to 6.0 ± 2.52, p <0.05)

When analyzing the obtained data, no statistically significant differences were found both between the subgroups receiving the anti-S100 + anti-eNOS SMD preparation in combination with various treatment options at different stages of Parkinson's disease, and between similar subgroups receiving the anti-S100 SMD drug.

Table 3 UPDRS Swab-England Questionnaire Beck Depression Questionnaire SMD anti-S100 + anti-eNOS before treatment 63.5 ± 9.77 65.0 ± 12.25 18.28 ± 14.33 SMD anti-S100 + anti-eNOS after treatment 49.16 ± 10.09 * # 78.33 ± 7.53 * 8.67 ± 7.99 SMD anti-S100 before treatment 70.125 ± 16.57 78.57 ± 13.45 12.0 ± 7.08 SMD anti-S100 after treatment 66.75 ± 14.62 78.75 ± 12.46 6.0 ± 2.52 * * - p from ref. <0.05 # - p compared to control <0.05

Thus, a clinical study of the combined preparation of SMD anti-S100 + anti-eNOS showed a positive effect on the main manifestations of Parkinson's disease and a possible effect on anxiety-depressive disorders of patients. In addition, the high tolerance of the drug SMD anti-S100 + anti-eNOS was confirmed. Adverse events when taking the drug SMD anti-S100 + anti-eNOS were absent.

Example 4

To study the properties of the claimed drug in the treatment of psycho-organic syndrome, 300 mg tablets were used, impregnated with a pharmaceutical composition containing aqueous-alcoholic solutions (6 mg / tablet) of activated-potentiated forms of polyclonal affinity-purified rabbit antibodies to brain-specific protein S-100 (anti -S100) and to endothelial NO-synthase (anti-eNOS) in ultra low doses (RMA) obtained sverhrazvedeniem initial matrix solution (2.5 mg / ml) 100 ¹² 100 ³⁰ 100 ²⁰⁰ times the equivalence GOVERNMENTAL mixture of centesimal homeopathic dilutions C12, C30, C200 (SMD anti-S100 + anti-eNOS.

As a control group, we used a group of patients receiving the drug in the form of tablets soaked in an aqueous-alcoholic solution (6 mg / tablet) of an activated-potentiated form of polyclonal rabbit antibodies to brain-specific protein S-100, purified on antigen, in an ultra-low dose (SMD anti -S100) obtained by super-dilution of the initial matrix solution 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of hundred homeopathic dilutions C12, C30, C200.

The study included patients with a diagnosis of psycho-organic syndrome of post-traumatic origin. The psycho-organic syndrome is characterized by a weakening of memory, a decrease in intelligence, and incontinence of affects (the Walter-Buel triad).

By design, the study was an open, randomized comparative clinical study of the efficacy and safety of the study therapy in two parallel groups in the treatment of patients with psycho-organic syndrome of post-traumatic genesis (the first group of patients took the drug SMD anti-S100, the second group of patients took the drug SMD anti-S100 + anti -eNOS.

The study included 6 patients with a diagnosis of “Organo-Syndrome Syndrome” aged 35 to 90 years. The average age of the patients was 70.83 ± 21.95 years.

The study examined patients' compliance with the following inclusion and exclusion criteria.

Inclusion criteria

1. Patients with a diagnosis of “Post-traumatic encephalopathy with psycho-organic syndrome” or “Mixed genesis encephalopathy (vascular, post-traumatic) with psycho-organic syndrome”, confirmed by medical history, neurological examination and medical documentation.

2. Constancy of concomitant therapy for at least a month before Visit 1.

3. The lack of prerequisites for a change in neurological concomitant therapy throughout the observation period.

4. Lack of prerequisites for the appointment of immunomodulating agents over the next 6 months.

5. The level of education and the degree of understanding of the patient are sufficient to adequately communicate with the researcher and the coordinator of the study.

6. Patients are evaluated by the researcher as reliable, ready to perform all prescribed clinical visits, tests and procedures provided for by the protocol.

7. Patients have a valid home address.

Exclusion Criteria:

1. A history of surgical intervention on the brain.

2. A history of stroke.

3. A history of psychosis, bipolar disorder, or schizoaffective disorder.

4. Major depressive disorder according to the criteria for the depression module of the international neuropsychiatric mini-interview (MINI).

5. Factors / conditions of a medical or other nature that, in the opinion of the researcher, may affect the patient's test results in this study.

6. Answers “2A”, “2B”, “2B” or “3” in the heading “I” of the Beck Depression Questionnaire (active suicidal thinking with some intention to act, without a specific plan or active suicidal thinking with a specific plan and intent).

7. A history of autoimmune disease.

8. Acute liver damage or severe cirrhosis (Child-Pugh class C).

9. Unregulated thyroid dysfunction.

10. An uncompensated history of hypertension.

11. Serious or decompensated cardiovascular disease, liver, kidney disease, metabolic, respiratory or hematological disease, symptomatic peripheral vascular disease, or other medical or psychiatric condition that, in the opinion of the researcher, may affect the patient’s participation in the study or may lead to prolongation of hospitalization or re-hospitalization of the patient during the study.

12. Diseases and conditions that, according to the researcher, may interfere with the participation of the patient in the study.

13. Taking Impaz (SMD anti-eNOS) or Tenoten (SMD anti-S100) before taking the study.

14. Admission of antidepressants of any group, not excluding herbal and homeopathic medicines.

15. Reception of anxiolytics of any group, not excluding herbal and homeopathic preparations.

16. Reception of immunomodulators, not excluding herbal and homeopathic preparations.

17. Systemic treatment with steroids, for 1 month or less before Visit 1.

18. Vaccinations, including against the flu.

19. Participation in the study of Impaz (SMD anti-eNOS) or Tenoten (SMD anti-S100), if patients have taken at least one dose of the drug.

20. Participation in other clinical trials for 1 month before inclusion in this study.

21. Pregnancy, breast-feeding, the inability to use adequate contraception during the study and within a month after the last administration of the study drug.

22. The presence of allergies / intolerances to any of the components of the drugs used in the treatment, including lactose intolerance.

23. The use of drugs, antipsychotics, alcoholism, mental illness of the patient.

24. Patients are the staff of the center directly related to the study, and / or are members of the family of the staff of the research center directly related to the study. The concept of "family members" includes a spouse (s), parents, children, brothers (sisters) associated with both biological and legal kinship.

25. Participation in litigation or alleged compensation or participation in litigation, according to the researcher.

After determining whether the patient met the criteria for inclusion and exclusion of the protocol, the patients were randomized into two study groups: a group of patients receiving anti-S100 SMD (3 people, women 33.33%, men 66.66%, average age 71.33 ± 16.25 years), and a group of patients receiving SMD anti-S100 + anti-eNOS (3 people, women 66.66%, men 33.33%, average age 70.33 ± 30.66 years) .

During this study, 5 visits were made to the research center. The treatment phase lasted from Visit 1 to Visit 4 for 84 ± 5 days. Visit 4 (day 84 ± 5) was the first endpoint of the study, after which the follow-up phase began. The follow-up phase continued until Visit 5 (day 168 ± 5).

The safety analysis included data from all patients participating in the study (n = 6). During the entire period of observation of patients, good tolerability of the drug was noted. Adverse events were absent. All patients of the study groups completed the treatment within the time period established by the study protocol; there were no prematurely retired patients.

When assessing the effect of the drug SMD anti-S100 + anti-eNOS on the main neuropsychiatric manifestations of illness of the psychoorganic syndrome (neuropsychic NPI questionnaire, section severity), on the severity of concomitant distress of the caregiver of the patient (neuropsychic questionnaire NPI, section distress), the patient's daily life activity ( questionnaire ADS-ADL), as well as the patient’s cognitive function (MMSE questionnaire), an improvement was revealed in the main neuropsychic manifestations of the psycho-organic syndrome, expressed in statistical terms a decrease in the total score of the “severity” section of the neuropsychic NPI questionnaire (from 91.0 + 15.13 to 69.0 + 6.244, p <0.05), and the level of distress of the caregiver, expressed as a statistically significant decrease in the total score of the section “distress” »The neuropsychic questionnaire NPI (from 44.33 + 17.78 to 36.33 + 3.21, p <0.05) to Visit 4 (Table 4).

Moreover, the difference between the groups of patients in the total score of the section “severity” of the neuropsychic NPI questionnaire by the end of therapy was significant with a probability p <0.05.

Similar indicators in the group of patients receiving anti-S100 SMD did not show improvement trends.

Table 4 NPI (severity) NPI (distress) ADS-ADL MMSE SMD anti-S100 + anti-eNOS before treatment 91.0 + 15.13 44.33 + 17.78 42.66 + 4.93 22.33 + 3.21 SMD anti-S100 + anti-eNOS after treatment 69.0 + 6.244 * # 36.33 + 3.21 * 52.0 + 5.57 22.66 + 2.08 SMD anti-S100 before treatment 114.0 + 25.53 45.66 + 14.47 33.0 + 13.89 22.33 + 4.16 SMD anti-S 100 after treatment 99.66 + 18.0 49.0 + 17.05 31.66 + 10.6 9 23.0 + 4.36 * - p from ref. <0.05 # - p compared to control <0.05

Thus, a clinical study of the combined preparation of SMD anti-S100 + anti-eNOS showed a positive effect on the main neuropsychic manifestations of PIC and a possible effect on the cognitive functions of patients with PIC. In addition, high tolerance to the drug has been confirmed. Adverse events when taking the drug were absent.

Example 5

In the following study, we studied the effectiveness of activated-potentiated forms of polyclonal affinity-purified rabbit antibodies on antigen for brain-specific protein S-100 (anti-S100) and endothelial NO synthase (anti-eNOS) in ultra-low doses (SMD) obtained by super-dilution of the initial matrix solution (with a concentration of 2.5 mg / ml) in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of hundreds of homeopathic dilutions C12, C30, C200 (SMD anti-S100 + anti-eNOS) in the treatment of ischemic stroke in rats caused by photothrombosis prefrontal cortex g tin brain.

Acute cerebrovascular accident (stroke) is the third leading cause of death in developed countries and is one of the leading causes of disability (Gusev E.I., 2003; Janardhan V., Qureshi A.I., 2004).

The model of photoinduced thrombosis meets almost all the requirements for experimental models of focal cerebral ischemia. The technique developed by Watson (Watson B. et al., 1985) is based on the fact that under the influence of light with a wavelength of 560 nm on the photosensitized Bengal rose dye introduced into the bloodstream, active oxygen forms are formed, under the influence of which the adhesiveness increases endothelial cells and platelets, and blood clots form that block the lumen of the vessels. The technique for reproducing ischemic pathology using photoinduced thrombosis is technically simple and as close as possible to the clinical forms of ischemic cerebral infarction. The great advantage of this model over others is that it is non-invasive, i.e. does not require craniotomy, and, thus, more accurately reproduces the clinical pathology that occurs with thrombosis of cerebral vessels.

In a study of the effectiveness of the SMD anti-S100 + anti-eNOS in rats with ischemic stroke caused by photothrombosis of the prefrontal cortex, 37 Wistar male rats were used (weight 150-180 g; 2-3 months). Bilateral focal ischemic damage in the prefrontal cortex of rats was caused by the method of photochemical thrombosis according to Watson (Watson B. D. et al., 1985) modified by I.V. Viktorova (Romanova G.A. et al., 1998). Rats (n = 37) under anesthesia (chloral hydrate 300 mg / kg, ip) were injected with a 3% dye solution (pink Bengal 40 mg / kg) into the jugular vein. Using a 3 cm diameter optical fiber, a light beam from a halogen lamp (24 V, 250 W) was brought to the surface of the skull above the frontal cortex of the left and right hemispheres of the brain, inducing photothrombosis. False-operated rats (n = 6) under the same conditions performed all the above manipulations, except for the introduction of dye and irradiation with light from a halogen lamp. The intact group included 6 rats.

To rats with photothrombosis, distilled water (control-photothrombosis, n = 12), SMD anti-S100 (n = 7) or SMD anti-S100 + anti-eNOS (n = 6) was injected intragastrically within 5 days before the induction of stroke and 9 days after ) at a dose of 5 ml / kg. On the 8th day after the operation (or false operation), to assess the learning ability and memory in all rats, the test of conditioned passive avoidance reflex (passive avoidance reaction) was performed. Rats were placed in a facility, which consists of an illuminated area and a darkened chamber connected to it, onto the floor of which an electric pain stimulation of 0.45 mA is applied, as a result of which the normally preferred darkened chamber became dangerous. The next day we tested the development of a conditioned reflex of passive avoidance. For this, rats were placed on a lighted area and the latent period of the first entry into the dark chamber was recorded. If the rat did not enter the dark chamber for a long time, it was concluded that it remembers the danger (electric pain irritation). The longer the latent period of entry into the camera, the better the safety of memory, which is impaired after a stroke caused by photothrombosis.

On the 9th day after the operation, the volume of the stroke zone in a morphometric study was estimated in some rats of the experimental groups.

In rats of the control group, photothrombosis led to the formation of an extensive zone of stroke and, as a result, to memory impairment - to a decrease in passive avoidance reaction by 9.6% compared to intact rats and by 22.9% compared to false-operated ones (Table 5). The introduction of SMD anti-S100 reduced the area of the stroke by 42.2% and improved memory by 14.0% compared with the control-photothrombosis. The introduction of SMD anti-S100 + anti-eNOS was more effective: in this group, the stroke focus was 44.0% less, and the reproduction of the conditioned reflex was 33.4% better than in the control-photothrombosis group.

Thus, the use of the complex preparation of SMD anti-S100 + anti-eNOS was more effective than the introduction of a single drug SMD anti-S100.

Table 5 The volume of the stroke, mm ³ ; number of animals Latent period passive avoidance reaction, sec; number of animals Intact - 135.8 ± 28.8; n = 6 Falsely operated - 159.3 ± 18.7; n = 6 Photothrombosis Control 3.41 ± 0.5; n = 9 122.8 ± 20.9; n = 12 Photothrombosis + SMD anti-S100 1.97 ± 0.6; n = 4 140.0 ± 26.5; n = 7 Photothrombosis + SMD anti-S100 + anti-eNOS 1.91 ± 0.5; n = 4 163.8 ± 16.2; n = 6

Claims (18)

1. A neurotropic drug containing an activated-potentiated form of antibodies to the brain-specific protein S-100, characterized in that it is made in the form of a pharmaceutical composition and contains an activated-potentiated form of antibodies to endothelial NO synthase as an additional reinforcing component. 2. The neurotropic drug according to claim 1, characterized in that the activated-potentiated form of antibodies to endothelial NO synthase and the activated-potentiated form of antibodies to brain-specific protein S-100 are used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution, activity which is due to the process of successive multiple dilutions in an aqueous or hydroalcoholic solvent in combination with an external mechanical action - vertical shaking of each dilution Niya. 3. The neurotropic drug according to claim 1 or 2, characterized in that the pharmaceutical composition is made in solid dosage form and contains an effective amount of granules of a neutral carrier saturated with a mixture of aqueous or aqueous-alcoholic solutions of an activated-potentiated form of antibodies to endothelial NO synthase and an activated-potentiated form of antibodies to the brain-specific protein S-100; and pharmaceutically acceptable additives. 4. The neurotropic drug according to claim 1 or 2, characterized in that aqueous or aqueous-alcoholic solutions of activated-potentiated forms of antibodies to endothelial NO synthase and to brain-specific protein S-100 are obtained by repeated serial dilution and external exposure - vertical shaking each dilution from matrix solutions of affinity purified antibodies to endothelial NO synthase and to brain-specific protein S-100 with a concentration of 0.5 ÷ 5.0 mg / ml 5. The neurotropic drug according to claim 1 or 2, characterized in that each of the components based on antibodies is used as a mixture of various, mainly hundred, dilutions according to homeopathic technology. 6. The neurotropic drug according to claim 3, characterized in that the pharmaceutically acceptable additives include lactose, microcrystalline cellulose and magnesium stearate. 7. The neurotropic drug according to claim 1 or 2, characterized in that the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat organic diseases of the nervous system . 8. The neurotropic drug according to claim 7, characterized in that the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat Parkinson's disease. 9. The neurotropic drug according to claim 1 or 2, characterized in that the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat the psycho-organic syndrome. 10. The neurotropic drug according to claim 1 or 2, characterized in that the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat cerebrovascular diseases. 11. The neurotropic drug of claim 10, characterized in that the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat acute cerebrovascular accident - stroke . 12. The neurotropic drug according to claim 1 or 2, characterized in that the pharmaceutical composition containing the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat encephalopathies of various origins. 13. A method of treating organic diseases of the nervous system, psycho-organic syndrome and encephalopathies of various genesis by introducing into the body a neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100, characterized in that the activated-potentiated form of antibodies is additionally simultaneously and simultaneously administered. to endothelial NO synthase. 14. The method according to item 13, characterized in that the activated-potentiated form of antibodies to endothelial NO synthase and the activated-potentiated form of antibodies to brain-specific protein S-100 are used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution of each component, activity which is due to the process of repeated dilution in an aqueous or hydroalcoholic solvent in combination with an external mechanical action - vertical shaking of each dilution. 15. The treatment method according to item 13 or 14, characterized in that use is made of a mixture of different dilutions of antibodies to endothelial NO synthase in combination with a mixture of different dilutions of antibodies to brain-specific protein S-100, prepared as a single drug - one dosage form cooked by homeopathic technology. 16. The treatment method according to item 13 or 14, characterized in that the neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat Parkinson's disease. 17. The treatment method according to item 13 or 14, characterized in that the neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat cerebrovascular diseases. 18. The treatment method according to item 13 or 14, characterized in that the neurotropic drug based on the activated-potentiated form of antibodies to the brain-specific protein S-100 and the activated-potentiated form of antibodies to endothelial NO synthase is used to treat acute cerebrovascular accident - a stroke.