RU2533224C2 - Method of treating psychoactive substance dependence, alcohol and nicotine addiction and medication for treatment of psychoactive substance dependence, alcohol and nicotine addiction - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: present group of inventions refers to medicine, namely to psychiatrics and addictology, and concerns treating psychoactive substance dependence, particularly alcohol and nicotine addiction. That is ensured by administering a therapeutic agent containing an activated potentiated form of antibodies against the brain-specific S-100 protein and an activated potentiated form of antibodies to a human cannabinoid receptor, more preferentially to the human cannabinoid receptor type I.

EFFECT: method provides suppressing both physical and mental alcohol and nicotine addiction.

11 cl, 1 ex

Description

The invention relates to medicine and can be used to treat addiction to psychoactive substances, including alcoholism and smoking.

The prior art neurotropic drug for the treatment of alcoholism based on the activated form of ultra-low doses of antibodies to the brain-specific protein S-100 (RU 2156621 C1, A61K 39/395, 09/27/2000). However, this drug does not in all cases of treatment for addiction to a narcotic substance provide sufficient therapeutic efficacy.

The invention is directed to the creation of a medicinal product that provides an increase in the effectiveness of the treatment of dependence, mainly on alcohol, nicotine and other psychoactive substances, including psychotropic drugs.

The solution to this problem is provided by the fact that in the method of treating dependence on psychoactive substances, mainly alcoholism and smoking, by introducing into the body an activated potentiated form of antibodies to the brain-specific protein S-100, according to the invention, an activated potentiated form of ultra-low doses of affinity-purified is additionally simultaneously combined. antibodies to a human cannabinoid receptor, preferably a human cannabinoid receptor of type I.

In this case, the activated potentiated form of antibodies to the brain-specific protein S-100 and the activated potentiated form of antibodies to the human cannabinoid receptor of type I 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 multiple sequential dilutions in aqueous or aqueous-alcoholic solvent in combination with external mechanical action - vertical shaking of each dilution.

Moreover, they use a mixture of different dilutions of antibodies to the brain-specific protein S-100 prepared in the form of a single drug - a single dosage form in combination with a mixture of different dilutions of antibodies to the human type I cannabinoid receptor prepared using homeopathic technology.

The solution to this problem is also provided by the fact that the drug for the treatment of dependence on psychoactive substances, mainly alcoholism and smoking, 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 enhancing component potentiated form of antibodies to human cannabinoid receptor, preferably to human cannabinoid receptor I type.

In this case, the activated potentiated form of antibodies to the brain-specific protein S-100 and the activated potentiated form of antibodies to the human cannabinoid receptor of type I 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 serial dilutions in an aqueous or aqueous-alcoholic solvent in combined with external mechanical action - vertical shaking of each dilution.

In addition, the pharmaceutical composition may be in solid dosage form that contains an effective amount of a neutral carrier granule saturated with a mixture of an activated potentiated form of antibodies to the brain-specific protein S-100 and an activated potentiated form of antibodies to a human cannabinoid receptor, and pharmaceutically acceptable additives that include lactose, microcrystalline cellulose and magnesium stearate.

In this case, aqueous or aqueous-alcoholic solutions of activated potentiated forms of antibodies to the brain-specific protein S-100 and to the human cannabinoid receptor are obtained by repeated serial dilutions in combination with an external action - vertical shaking of each dilution from matrix solutions of affinity-purified antibodies to the brain-specific protein S-100 and to the human cannabinoid receptor with a concentration of 0.5-5.0 mg / ml.

In addition, each of the components of ultra-low doses of affinity-purified antibodies is used in the form of a mixture of various, mainly hundreds, dilutions prepared by homeopathic technology.

In this case, the drug contains active ingredients in a volume ratio of 1: 1, with each component being used as a mixture of three appropriate matrix solutions, diluted, respectively, 100 ¹² , 100 ³⁰ , and 100 ²⁰⁰ times, which is equivalent to hundreds of dilutions of C12, C30 , C200, prepared according to homeopathic technology.

The claimed pharmaceutical composition is recommended to be taken, preferably, 1-2 tablets 2-6 times a day.

The claimed technical solution can be used both for the treatment of mental attraction to psychoactive substances, including alcohol and tobacco smoking, and for the treatment of psychosomatic disorders associated with deprivation of a psychoactive substance - withdrawal conditions.

In addition, the claimed drug can be used in combination therapy with other neuropsychotropic drugs used to treat addictive conditions.

In the treatment of dependence on psychoactive substances, alcoholism and tobacco smoking, separate but combined and simultaneous use (administration to the body) in the form of two separately prepared drugs, both in the form of solutions and in solid dosage forms (tablets), each of which contains activated the potentiated form of the ultra-low doses of affinity-purified antibodies to the brain-specific protein S-100 and, accordingly, the activated potentized form of the ultra-low doses of affinity purified antibodies to the cannabinoid recipe human torus.

The proposed combination of activated potentiated forms of antibodies to the brain-specific protein S-100 and to the human type I cannabinoid receptor in a pharmaceutical composition (i.e., antibodies to the brain-specific S-100 protein and to the human type I cannabinoid receptor prepared using the homeopathic technology of potentiation by multiple serial dilution in combination with external exposure - vertical shaking of each dilution (see, for example, V. Schwabe "Homeopathic medicines", M., 1967, p. 14-29), which are active in pharmacological models and / or clinical methods of treating dependence on psychoactive substances, mainly alcoholism and smoking) provides an unexpected synergistic therapeutic effect, confirmed on adequate (valid) experimental models and clinical studies. At the same time, the claimed drug modulates the system of positive emotional support, thereby reducing the severity of attraction to psychoactive substances, including alcoholism and nicotinism, normalizes the balance of neurotransmitters in the central nervous system, improves neuroprotective activity, increases brain resistance to toxic effects, improves integrative activity , helps to eliminate cognitive impairment, increases mental performance, normalizes somatovegetative symptoms in withdrawal conditions. As a result of this, irritability is eliminated, mood improves, internal discomfort is eliminated, thoughts of nicotine or alcohol are eliminated, working capacity, appetite is eliminated, sweating, tachycardia and general weakness are eliminated, as well as other affective, ideative and asthenovegetative disorders; quality of life improves. At the same time, the claimed neurotropic drug does not have a sedative, muscle relaxant effect, is not addictive, does not have drug dependence, does not have narcogenic potential and other side effects and has an extended therapeutic range in the treatment of dependence on psychoactive substances.

In addition, the claimed drug expands the arsenal of drugs designed to treat addiction to psychoactive substances.

The drug is prepared as follows.

To prepare 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, M., "Medicine", 1987, S. 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 human type I cannabinoid receptor. As a result of this procedure, monospecific antisera with a high content of antibodies are obtained, which are used to obtain activated potentiated forms. 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 type I cannabinoid receptor, which are used as a matrix (primary) solution with a concentration of 0.5-5.0 mg / ml for the 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 primary matrix solution of antibodies diluted, respectively, 100 ¹² , 100 ³⁰ , and 100 ²⁰⁰ times, which corresponds to hundreds of dilutions of C12, C30 and C200 prepared according to homeopathic technology. When performing the claimed drug in solid dosage form, a mixture of these components is applied to lactose.

Preferred for the preparation of the claimed drug is the use of polyclonal antibodies to the brain-specific protein S-100 and to the human cannabinoid receptor type I, which can be obtained by immunization of rabbits as follows.

Example 1

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

For immunization of rabbits, an adjuvant and a whole human type I cannabinoid receptor molecule are used as an immunogen (antigen):

or, preferably, a polypeptide fragment of a human type I cannabinoid receptor, selected, for example, from the following group:

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 the cannabinoid receptor type I from an antiserum of a person, type 1 absorption is performed on the solid phase 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 the human type I cannabinoid receptor, which is located on an insoluble matrix, followed by elution with concentrated salt solutions.

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. Starostin, S. M. Sviridov, Neurospecific protein S-100, Successes in modern biology, 1977. , t.5, p. 170-178; book of MB Stark, Brain-specific proteins / antigens / and neuron functions, M., "Medicine", 1985; p.12-14, isolated from tissue of the brain of a bull 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 Dal.

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 nervous tissue, but does not react with extracts of heterologous organs, and forms a single precipitation peak with both pure S-100 protein and nervous tissue extract, which indicates the monospecificity of the obtained antiserum.

Thus obtained buffer solutions of each component of polyclonal antibodies to the brain-specific protein S-100 and to and the human type I cannabinoid receptor with a concentration of 0.5 ÷ 5.0 mg / ml, preferably 2.0 ÷ 3.0 mg / ml, are used in as matrix (primary) solutions for the subsequent preparation of activated potentiated forms of the drug.

The activated potentiated form of each component is prepared by uniformly decreasing the concentration as a result of sequential dilution of 1 part of the above matrix solution in 9 parts (for decimal dilution D), or in 99 parts (for hundredth dilution C), or in 999 parts (for thousandth dilution M) neutral solvent with multiple vertical shaking ("dynamization") of each dilution obtained and the use of separate containers for each subsequent dilution to obtain the desired potency and - the multiplicity of homeopathic dilution method (see, eg, W. Schwabe "Homeopathic medicines", M., 1967, s.14-29.).

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

For example, for the preparation of the 12th hundredth C12 dilution, one part of the matrix solution of antibodies to the brain-specific protein S-100 (or to the human cannabinoid receptor type I) with a concentration of 3.0 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 sequentially in different containers 12 times of the 1st part of the initial matrix solution of antibodies to the brain-specific protein S-100 with a concentration of 3.0 mg / ml in 99 parts of a neutral solvent , a solution prepared by diluting the matrix solution 100 to ¹² times. Similar operations with the appropriate dilution ratio are carried out to obtain dilutions of C30 and C200.

When using as a biologically active liquid component a mixture of various, mainly hundred, dilutions of the active substance prepared according to homeopathic technology, each component of the composition (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, in accordance with the composition of the mixture, make one part of each component in one container and mix with the required amount of solvent (respectively, with 97 parts for centesimal 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 prepared by homeopathic technology.

It is possible to use the active substance in the form of a mixture of other various dilutions, for example, decimal and / or hundredths (D20, C30, C100 or C12, C30, C50, etc.) prepared according to homeopathic technology, the effectiveness of which is determined experimentally.

When potentiating instead of shaking in the process of decreasing the concentration, it is also possible to exert external influence by ultrasound, electromagnetic or other physical effect.

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 also be used in solid dosage form, which contains an effective amount of granules of a neutral carrier — lactose, saturated by soaking in a mixture of aqueous or aqueous-alcoholic solutions with an activated potentiated form of antibodies to the brain-specific protein S-100 and an activated potentiated form of antibodies to human cannabinoid receptor type I, and pharmaceutically acceptable additives, including, mainly, lactose, microcrystalline cellulose and magnesium and stearate.

To obtain a solid oral form of the claimed drug, a fluidized bed is installed (for example, Huttlin Pilotlab type manufactured by Huttlin GmbH) irrigation until saturation of granules of a neutral substance - lactose (milk sugar) with a particle size of 150-300 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 the human cannabinoid receptor type I, mainly in a ratio of 1 kg p alignment antibodies to 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 of not higher than 40 ° C. The estimated amount of 0.07-0.17 by weight of the solid oral form of the dried granules saturated with the activated potentiated form of antibodies is loaded into the mixer and mixed with microcrystalline cellulose introduced in an amount of 3.0-8.0 mass. parts of the total mass of the load - from the mass of solid oral form. Then, 25-45 masses are added to this mixture. parts of the total mass of the load of "unsaturated" pure lactose (to reduce the cost and some simplification and acceleration of the process without reducing the effectiveness of the therapeutic effect) and magnesium stearate in an amount of 0.8-1.2 mass. 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. The weight of the tablet may be 150 ÷ 500 mg. It is preferable to use tablets weighing 250–300 mg, which include an activated potentiated form of water-alcohol dilutions of a substance, the active substance of polyclonal rabbit antibodies to brain-specific protein S-100 and to type I cannabinoid receptor, purified on an antigen in an ultra-low dose prepared from a matrix solution diluted, respectively, in 100 ¹² , in 100 ³ in 100 ²⁰⁰ , which is equivalent to a mixture of hundred-percent dilutions of C12, C30 and C200 prepared according to homeopathic technology.

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

Claims (11)

1. A method of treating dependence on alcoholism and smoking by introducing into the body an activated potentiated form of antibodies to the brain-specific protein S-100, characterized in that an additional reinforcing component in the form of an activated potentiated form of antibodies to the human cannabinoid receptor is additionally simultaneously and simultaneously administered. 2. The method according to claim 1, characterized in that as an additional reinforcing component using an activated potentiated form of antibodies to cannabinoid receptor type I person. 3. The method according to claim 1 or 2, characterized in that the activated potentiated form of antibodies to the brain-specific protein S-100 and the activated potentiated form of antibodies to the cannabinoid receptor of human type I are used in the form of an activated potentiated aqueous or aqueous-alcoholic solution of each component obtained in the process of successive multiple dilutions in an aqueous or hydroalcoholic solvent in combination with an external mechanical action - shaking of each dilution. 4. The treatment method according to claim 1 or 2, characterized in that they use a mixture of different homeopathic dilutions of antibodies to the brain-specific protein S-100, prepared in the form of a single drug, in combination with a mixture of different dilutions of antibodies to the cannabinoid receptor type I person prepared by homeopathic technology. 5. A drug for the treatment of addiction to alcoholism and smoking, 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 additionally contains an enhancing component in the form of an activated potentiated form of antibodies to the human cannabinoid receptor. 6. The drug according to claim 5, characterized in that as an additional reinforcing component using an activated potentiated form of antibodies to the cannabinoid receptor type I person. 7. The drug according to claim 6, characterized in that the activated potentiated form of antibodies to the brain-specific protein S-100 and the activated potentiated form of antibodies to the cannabinoid receptor of human type I are used in the form of an activated potentiated aqueous or aqueous-alcoholic solution, the activity of which is due to the process successive multiple dilutions in an aqueous or hydroalcoholic solvent in combination with an external mechanical action - shaking of each dilution. 8. The drug according to claim 5 or 7, characterized in that the pharmaceutical composition is in solid dosage form and contains an effective amount of granules of a neutral carrier and an activated potentiated form of antibodies to the human specific brain protein S-100 and an activated potentiated form of antibodies to the human cannabinoid receptor of a neutral carrier and pharmaceutically acceptable additives. 9. The drug according to claim 5 or 7, characterized in that the aqueous or aqueous-alcoholic solutions of activated potentiated forms of antibodies to the brain-specific protein S-100 and to the human cannabinoid receptor are obtained by multiple serial dilutions in combination with the external action of each dilution from matrix solutions of antibodies to the brain-specific protein S-100 and to the human cannabinoid receptor with a concentration of 0.5 ÷ 5.0 mg / ml 10. The drug according to claim 5 or 7, characterized in that each of the components based on activated potentiated forms of antibodies to the brain-specific protein S-100 and to the human cannabinoid receptor is used as a mixture of various, mainly hundred, dilutions obtained by homeopathic technology . 11. The drug of claim 8, characterized in that the pharmaceutically acceptable additives include lactose, microcrystalline cellulose and magnesium stearate.