RU2532323C2 - Therapeutic agent for treating pathological syndrome and method of treating functional bowel disorders - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: therapeutic agent containing activated-potentiated forms of anti-histamine, anti-tumour necrosis factor alpha (anti-TNF - α) and anti-S-100 brain specific antibodies is administered.

EFFECT: treating functional bowel disorders by ensuring the spasmolytic action and normalising the motor-evacuation function of the intestine.

11 cl, 4 ex, 4 tbl

Description

The invention relates to medicine and can be used to treat functional bowel disorders, including irritable bowel syndrome and impaired intestinal motor-evacuation function.

Functional diseases of the intestine should be understood as such diseases of the intestine when the disorder of its function does not occur as a result of organic damage, but as a result of a violation of the nervous and humoral regulation of the digestive tract (organic lesions of the intestine are either absent or play a secondary role). Functional intestinal diseases are characterized by the absence of morphological changes (which could explain the existing clinical symptoms) and their relationship, firstly, with increased excitability of motility, secondly, with sensory hypersensitivity and, thirdly, with an inadequate response of internal organs to signals the central nervous system when exposed to psychosocial factors. Functional intestinal diseases are the most common type of functional pathology of the gastrointestinal tract and are observed in 40-70% of patients with a gastroenterological profile. The formation of functional intestinal disorders is influenced by genetic factors, environmental factors, psychosocial factors, visceral hypersensitivity and infections. Functional bowel diseases form part of a large group of diseases of the gastrointestinal tract related to functional pathology and, according to the classification of functional bowel diseases (Roman Consensus, 1999), include such clinical conditions as irritable bowel syndrome, functional flatulence, functional constipation, functional diarrhea and non-specific functional bowel disorders. In the pathogenesis of diseases, an essential role is played by violations of the motor function of the stomach and intestines. A feature of patients with functional bowel diseases is an increase in motor and sensory reactions, the appearance of abdominal pain in response to stress. The complex of functional intestinal disorders includes complaints of abdominal pain (usually decreasing after a bowel movement), flatulence, rumbling, a feeling of incomplete bowel movement, mandatory urge to defecate, constipation, diarrhea or their alternation. Clinical features common to all functional diseases of the gastrointestinal tract: long-term (usually long-term) course of the disease without noticeable progression; the diversity of the clinical picture (a combination of abdominal pain, dyspeptic disorders and intestinal dysfunctions with headaches such as migraines, sleep disturbances, coma when swallowing, dissatisfaction with inspiration, inability to sleep on the left side, frequent urination, various vasospastic reactions and other autonomic disorders) ; the volatile nature of complaints; connection worsening well-being with psycho-emotional factors.

Irritable bowel syndrome (IBS) is one of the most common functional bowel diseases, which is recorded, as recent observations show, in almost all countries of the world. The prevalence of IBS in most countries of the world averages 20%, varying, according to various studies, from 9 to 48%. The peak incidence occurs in young working age - 30-40 years. The ratio of women to men ranges from 1: 1 to 2: 1. Among men of “problem” age (after 50 years), IBS is as common as among women. The average age of patients is 24-41 years.

The etiology and pathogenesis of IBS are complex and not fully known. Most researchers agree on the important role of psychoemotional stress in the development of IBS. Depending on the leading symptom, three variants of the course of IBS are distinguished: with predominant abdominal pain and flatulence, with predominant diarrhea and with predominant constipation. Until 1988, IBS was described under various names, such as spastic colitis, mucous colic, nervous diarrhea, irritable colon, functional intestinal distress syndrome, and others. These names reflected various symptoms of the disease and did not reflect a common understanding of the problem. In 1988, in Rome, the International Group for the Study of Functional Pathology of the Gastrointestinal tract (GIT) for the first time officially approved the term “irritable bowel syndrome”, gave its definition and developed the criteria for diagnosis, which later became known as the “Roman criteria for IBS”. In 1999, the criteria were supplemented and adopted by the "Roman criteria of IBS II", and in 2006 - the "Roman criteria of IBS III". According to the “Roman Criteria III”, IBS is a stable combination of functional disorders, manifested in the presence of recurring abdominal pain or discomfort, noted for at least 3 days during the month over the past 3 months and combined with two of the following three symptoms: pain decrease after the act of defecation; pain is accompanied by a change in stool frequency; pain is accompanied by a change in the consistency of the stool; these symptoms should be noted in patients within the last 3 months; their total duration should be at least 6 months; discomfort should be understood as unpleasant sensations that are not described by patients as pain; for pathophysiological and clinical studies, it is recommended to select patients in whom pain and discomfort are noted at least 2 days a week.

A medicine is known from the prior art for the treatment of erosive and inflammatory diseases of the gastrointestinal tract based on activated forms of ultra-low doses of antibodies to morphine and activated forms of ultra-low doses of antibodies to histamine (RU 2197266 C1, A61K 39/395, 01.27.2003). However, this drug may not in all cases provide sufficient therapeutic efficacy for the treatment of functional bowel disorders, including irritable bowel syndrome and impaired intestinal motor-evacuation function.

The invention is aimed at increasing the effectiveness of the treatment of functional bowel disorders, including irritable bowel syndrome and impaired intestinal motor-evacuation function.

The solution to this problem is provided by the fact that the drug for the treatment of pathological syndrome, including for the treatment and prevention of functional intestinal disorders, containing the activated-potentiated form of antibodies to histamine, according to the invention, is made in the form of a pharmaceutical composition and contains as additional (enhancing) components of the activated-potentiated form of antibodies to the tumor necrosis factor - alpha (TNF-α) and the activated-potentiated form of antibodies to brain-specific eskomu protein S-100.

The indicated functional disorders of the gastrointestinal tract can be, in particular, irritable bowel syndrome, impaired intestinal motor-evacuation function, flatulence, functional constipation, functional diarrhea, nonspecific functional intestinal disorders, etc.

The activated-potentiated form of antibodies to histamine, the activated-potentiated form of antibodies to TNF-α and the activated-potentiated form of antibodies to the brain-specific protein S-100 are used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution obtained by sequential multiple dilution in aqueous or a water-alcohol solvent and an intermediate external mechanical effect - vertical shaking.

The pharmaceutical composition of the claimed drug can be made in solid dosage form, which contains an effective amount of granules of a neutral carrier saturated with a mixture of aqueous or aqueous-alcoholic solutions of the activated-potentiated form of antibodies to histamine, the activated-potentiated form of antibodies to TNF-α and the activated-potentiated forms of antibodies to the brain-specific protein S-100; and pharmaceutically acceptable additives. In this case, pharmaceutically acceptable additives include, for example, lactose, microcrystalline cellulose and magnesium stearate.

Aqueous or aqueous-alcoholic solutions of activated-potentiated forms of antibodies to histamine, to TNF-α and to brain-specific protein S-100 can be obtained by repeated serial dilution and intermediate external exposure from matrix solutions of affinity-purified antibodies to histamine, to TNF-α and to the brain-specific protein S-100 with a concentration of 0.5 ÷ 5.0 mg / ml.

Moreover, each of the components of ultra-low doses of affinity-purified antibodies is used in the form of a mixture of various, mainly hundred, homeopathic dilutions.

Preferably, the activated-potentiated form of antibodies to histamine, the activated-potentiated form of antibodies to TNF-α and the activated-potentiated form of antibodies to the brain-specific protein S-100 in an ultra-low dose of each component prepared from a matrix solution diluted in 100 ¹² , in 100 ³⁰ , in ^100,200 , which is equivalent to a mixture of hundred-percent homeopathic dilutions of C12, C30, C200.

The solution to this problem can also be achieved by the fact that in the method of treating functional intestinal disorders by introducing into the body a medicine containing an activated - potentiated form of anti-histamine antibodies, according to the invention, an activated-potentiated form of ultra-low doses of affinity-purified antibodies to TNF is additionally combined and administered -α and the activated-potentiated form of antibodies to the brain-specific protein S-100.

In this case, a mixture of various homeopathic dilutions of antibodies to histamine combined with a mixture of various homeopathic dilutions of antibodies to TNF-α and a mixture of various homeopathic dilutions of antibodies to brain-specific protein S-100 can be prepared in the form of a single drug - a single dosage form.

The claimed drug can be used for the treatment and prevention of patients with functional bowel disorders, including irritable bowel syndrome, impaired intestinal motor-evacuation function, etc.

In addition, the claimed drug, as a complex pharmaceutical composition, may contain active ingredients in a ratio of 1: 1: 1, with each component being used as a mixture of three appropriate matrix solutions diluted 100 ¹² , 100 ³⁰ , and 100 ²⁰⁰ times, which is equivalent to hundreds of homeopathic dilutions of C12, C30, C200.

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

In the treatment of functional intestinal disorders, separate use is possible in the form of three 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 histamine, an activated-potentiated form ultra-low doses of affinity-purified antibodies to TNF-α and, accordingly, an activated-potentiated form of ultra-low doses of affinity-purified antibodies to brain-specific protein S-100.

The proposed combination of activated-potentiated forms of antibodies to histamine, to TNF-α and to brain-specific protein S-100 in a pharmaceutical composition (i.e., forms of antibodies to histamine, to TNF-α and to brain-specific protein S-100, prepared according to homeopathic technology potentiation by repeated serial dilution and intermediate external action - vertical shaking, which are active in pharmacological models and / or clinical methods of treatment of functional intestinal disorders) This means that an unexpected synergistic therapeutic effect is achieved, which consists in a more pronounced therapeutic effect in the treatment of functional intestinal disorders, in particular, irritable bowel syndrome, impaired intestinal motor-evacuation function, etc., as compared to the action exerted by individual components, which is confirmed by adequate experimental models and clinical studies, which consists in normalizing the nervous and humoral regulation of bowel function, reducing intestinal hypersensitivity of the colon receptors to stretching, which ensures the restoration of intestinal motility disorders, relief of the sensation of bloating and overfilling of the stomach, reducing the severity of pain. In this case, smooth muscles relax, a decrease in the tone of the wall of the gastrointestinal tract (GIT), a decrease in intraluminal pressure, normalization of the stool consistency, its frequency and associated symptoms (relief of imperative urges, tenesmus, feelings of incomplete bowel movement, additional efforts during defecation, etc. .).

Thus, the drug contains three components that, in combination, have a multifactorial positive (therapeutic) effect on various manifestations of impaired intestinal activity and their claimed joint use in the form of a pharmaceutical composition is accompanied by normalization of neurohumoral regulation and gastrointestinal activity.

In addition, the claimed drug expands the arsenal of drugs intended for the treatment and prevention of functional bowel disorders.

The drug is prepared mainly as follows.

To prepare a homeopathically 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. For experimental studies, antibodies prepared by order of a specialized pharmaceutical company can be used.

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. To do this, according to a specially developed scheme, the animals are given a series of injections with the substance required in accordance with the invention - an antigen: histamine, TNF-α and brain-specific protein S-100. As a result of this procedure, a monospecific antiserum with a high antibody content is obtained, which is used to obtain the activated-potentiated form. If necessary, the antibodies present in the antiserum are purified, for example, by affinity chromatography, using fractionation by salt precipitation or ion exchange chromatography.

Polyclonal antibodies to histamine, which is a biogenic amine (4- (2-aminoethyl) -imidazole or beta-imidazolyl-ethylamine with the chemical formula C ₅ H ₉ N ₃ ), are prepared using an adjuvant as an immunogen (antigen) for immunizing rabbits commercially available histamine dihydrochloride.

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 13000 g 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 anti-histamine antibodies from the 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 removal of 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 histamine, which is located on an insoluble matrix, followed by elution with concentrated salt solutions.

Thus obtained buffer solution of polyclonal rabbit antibodies to histamine, purified on 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.

Obtaining polyclonal antibodies to the tumor necrosis factor-alpha (TNF-α) can be performed according to the above method of obtaining polyclonal antibodies to histamine using a whole tumor necrosis factor-alpha molecule of the following sequence:

1 MSTESMIRDV ELAEEALPKK TGGPQGSRRC LFLSLFSFLI VAGATTLFCL LHFGVIGPQR

61 EEFPRDLSLI SPLAQAVRSS SRTPSDKPVA HVVANPQAEG QLQWLNRRAN ALLANGVELR

121 DNQLVVPSEG LYLIYSQVLF KGQGCPSTHV LLTHTISRIA VSYQTKVNLL SAIKSPCQRE

181 TPEGAEAKPW YEPIYLGGVF QLEKGDRLSA EINRPDYLDF AESGQVYFGI IAL

It is possible to obtain polyclonal antibodies to the tumor necrosis factor alpha (TNF-α) using a polypeptide fragment of the tumor necrosis factor selected, for example, from the following sequences:

84-88:

PSDKP

93-97:

Vanpq

65-199:

RDLSLI SPLAQAVRSS SRTPSDKPVA HVVANPQAEG QLQWLNRRAN ALLANGVELR DNQLVVPSEG LYLIYSQVLF KGQGCPSTHV LLTHTISRIA VSYQTKVNLL SAIKSPCQRE TPEGAEAKPW YEPIYLGGV

77-93:

RSS SRTPSDKPVA HVV

32-54:

GGPQGSRRC LFLSLFSFLI VAGA

56-73:

IGPQR EEFPRDLSLI SPL

123-160:

QLVVPSEG LYLIYSQVLF KGQGCPSTHV LLTHTISRIA

176-190:

PCQRE TPEGAEAKPW

5-45:

SMIRDV ELAEEALPKK TGGPQGSRRC LFLSL

150-184:

VLLTHTISRIA VSYQTKVNLL SAIKSPCQRE TPEG

77-233:

VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGANGR

DNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKV NLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINR PDYLDFAESGQVYFGIIAL.

Obtaining antibodies to the brain-specific protein S-100. To prepare a potentiated form of antibodies - antisera to the brain-specific protein S-100, use the brain-specific protein S-100 isolated from the 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 D.

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

The resulting antiserum has a titer of 1: 500 -1: 1000.

Preferred for the preparation of the claimed drug is the use of polyclonal antibodies to histamine, to TNF-α and to brain-specific protein S-100, which as a matrix (primary) solution with a concentration of 0.5 ÷ 5.0 mg / ml (mainly 2, 0α3.0 mg / ml) is used for the subsequent preparation of the activated-potentiated form.

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 matrix solution in 9 parts (for decimal dilution) or in 99 parts (for hundredth dilution) or in 999 parts (for thousandth dilution) of a neutral solvent with multiple vertical shaking ("dynamization") of each dilution obtained and using separate containers for each subsequent dilution until the desired potency is obtained - cr dilution characteristics according to the homeopathic method (see, for example, V. 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 C12 dilution, one part of the mentioned 2.5 mg / ml histamine antibody matrix solution is diluted in 99 parts of a neutral aqueous or hydroalcoholic solvent (mainly 15% ethanol) and repeatedly (10 and more than once) 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 C 12. Thus, the 12th hundredth dilution of C 12 is a solution obtained by diluting successively in different containers 12 times of the 1st part of the initial matrix solution of antibodies to histamine with a concentration of 2 5 mg / ml in 99 parts of a neutral solvent, i.e. a solution prepared from a matrix solution diluted 100 to ¹² times. Similar operations with the appropriate dilution ratio are carried out to obtain a dilution of C30 and C200.

When using as a biologically active liquid component a mixture of various homeopathic, mainly hundred, dilutions of the active substance, each component of the composition (for example, C12, C30, C200) is prepared separately according to the technology described above to the penultimate dilution (respectively, to obtain C11, C29, C199) and then, in accordance with the composition of the mixture, one part of each component is added to one container and mixed with the required amount of a neutral aqueous or aqueous-alcoholic solvent (mainly but 50-70% ethyl alcohol) (respectively, with 97 parts for hundred percent dilution). In this case, an activated-potentiated form of antibodies to histamine is obtained in an ultra-low dose prepared from a matrix solution diluted in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ , which is equivalent to a mixture of hundred-percent homeopathic dilutions of C12, C30, C200.

It is possible to use the active substance in the form of a mixture of various different homeopathic ones, dilution, for example, decimal and or hundredths (D20, C30, C100 or C12, C30, C50, etc.), 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 ratio of 1: 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 - for example, lactose - saturated by soaking in a mixture of aqueous or aqueous-alcoholic solutions with an activated potentiated form of anti-histamine antibodies, an activated-potentiated antibody form TNF-α and the activated-potentiated form of antibodies to the brain-specific protein S-100, and pharmaceutically acceptable additives, including mainly lactose, microcrystalline cellulose and magnesium stearate.

To obtain a solid oral form of the claimed drug, a fluidized bed is installed (for example, type “Hiittlin Pilotlab” manufactured by Huttlin GmbH) irrigation until saturation of granules of a neutral substance - lactose (milk sugar) with a particle size of 150-300 μm, a pre-prepared aqueous or aqueous-alcoholic solution of activated-potentiated forms of antibodies to histamine, activated-potentiated forms of antibodies to TNF-α and activated-potentiated forms of antibodies to brain-specific protein S-100, mainly in the ratio of 1 kg of antibody solution 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 no higher than 40 ° C. The estimated amount of 0.17 ÷ 0.45 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 2 ÷ 5 mass. parts of the total mass of the load - from the mass of solid oral form. Then, 10 ÷ 25 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), magnesium stearate in an amount of 0.1 ÷ 0.3 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. After tabletting, 300 mg tablets are obtained, impregnated with an aqueous-alcoholic solution (3.0-6.0 mg / tablet) with an activated-potentiated form of antibodies to histamine, to TNF-α and to the brain-specific protein S-100 in an ultra-low dose of each component prepared from a matrix solution diluted in 100 ¹² , in 100 ³⁰ , in 100 ²⁰⁰ , which is equivalent to a mixture of hundred-percent homeopathic dilutions of C12, C30, C200.

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

In the following Examples 1-3, we studied the effectiveness of the claimed complex drug in the treatment of functional bowel disorders in the form of a composition containing antibodies to histamine, affinity purified on antigen, in an ultra-low dose obtained by super-dilution of the initial matrix solution (concentration 2.5 mg / ml) in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times (AT Gis), antibodies to S100 protein, affinity purified on antigen, in the ultra-low dose obtained by super-dilution of the initial matrix solution (concentration 2.5 mg / ml) in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times (AT S100) and en Ithel tumor necrosis factor - alpha, affinity purified on an antigen, in ultralow dose received sverhrazvedeniem initial matrix solution (concentration 2.5 mg / ml) 100 ¹² 100 ³⁰ 100 ²⁰⁰ time (AT TNF-α) (AT S100 + AT TNF-α + AT His). Comparison drug: anti-histamine antibodies affinity purified on antigen in an ultra-low dose obtained by super-dilution of the initial matrix solution (concentration 2.5 mg / ml) 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times (AT Gis),

Example 1

In a study of the effect of the claimed drug on the motor-evacuation function of the intestines of mice, 31 outbred male mice were used (weight 17.5-26.3 g, age 1.5-2 months), which were administered intragastrically for 5 days with distilled water (control 15 ml / kg, 10 mice), AT His (15 ml / kg, 11 mice) or AT S100 + AT TNF-α + AT His (15 ml / kg, 10 mice). The state of the motor-evacuation function of the intestine was studied by the method of “tags” (Coopman GP, Kermis HM Two methods to assess the gastrointestinal transit - time in mice // Z. Versuchstierk. 1977. Vol.19, No.5. P.298-303) . To do this, 1 hour after the last injection of the mice, a 10% suspension of activated charcoal prepared with 2% mucilage of potato starch in the amount of 0.5 ml / one mouse was injected into the digestive tract as mice. 10 minutes after the introduction of the “label”, the intestines were removed from the mice, stretched on a glass plate, the total length of the intestine and filled with charcoal was measured (the ratio of the length of the part of the intestine filled with activated charcoal to the total length was expressed as a percentage).

It was found that a preliminary course administration of AT S100 + AT TNF-α + AT Gis to healthy male mice at a dose of 15 ml / kg led to a statistically significant increase in the path length of activated carbon in the intestine by 1.3 and 1.2 times relative to the corresponding values AT gis groups and distilled water (control) (Table 1). Moreover, in this group, the ratio of the length of the coal-filled part to the total length of the intestine also exceeded the similar indicators in the AT Gis group (P <0.05) and the control group (P <0.05).

Table 1 The influence of the tested drugs on the motor-evacuation activity of the intestines of outbred male mice during the study using "carbon labels" Experimental group (number of animals) The length of the part of the intestine filled with coal per 1 mouse (M ± m), cm The length of the coal-filled part to the total length of the intestine (M ± m),% Diet water (n = 10) 25.4 ± 1.49 44.1 ± 2.77 AT Gis (n = 11) 24.1 ± 1.92 44.9 ± 3.65 AT S100 + AT TNF-α + AT His (n = 10) 31.1 ± 2.09 * # 54.4 ± 3.24 * # * - differences are significant relative to the control group (p <0.05) # - differences are significant relative to the AT Gis group (p <0.05)

Thus, it was shown that the complex preparation AT S100 + AT TNF-α + AT Gis enhances the motor-evacuation activity of the intestines of mice, that is, it has a moderate tonogenic effect, superior in efficiency to AT Gis.

Example 2

In a study of the effect of the claimed complex drug on the excretory function of the intestines of mice, 33 outbred male mice (weight 17.5-26.3 g, age 1.5-2 months) were used, which were administered intragastrically once with distilled water (control, 15 ml / kg ), AT His (15 ml / kg) or AT S100 + AT TNF-α + AT His (15 ml / kg). The state of the excretory function of the intestine was studied by the method of G.V.Obolentseva (Obolentseva G.V., Khadzhai Y.I. Pharmacological study of plantaglucide // Pharmacology and Toxicology. - 1996. - No. 4. - S.469-472). For this, the preparations were administered together with activated carbon at a dose of 10 mg / kg. The appearance of feces stained with black coal was considered a positive result. Measurements of the results were carried out after 3, 6, 24 hours from the start of the experiment. The degree of manifestation of the effect for each animal was determined in points: "+" - the appearance of the decorated colored feces; "++" - the appearance of soft colored feces; "+++" - the appearance of liquid stool. The laxative activity of the drugs was evaluated by the total score in the group and by the percentage of animals with a positive reaction. The effect of the tested drugs on the excretory function of the intestine in outbred male mice is shown in Table 2.

table 2 The influence of the tested drugs on the excretory function of the intestine in outbred male mice Monitoring group (number of animals) The magnitude of the effect (score /% of animals with reaction) 3 h 6 h 24 h Four times the introduction of drugs Control (n = 11) 22/100 18/100 7/55 AT Gis (n = 11) 18/100 18/100 6/55 AT S100 + AT TNF-α + AT His (n = 11) 25/100 24/100 6/55

A slight increase in peristalsis occurred already in the first 3 hours after the administration of AT S100 + AT TNF-α + AT Gis: 25 points vs 18 points in the AT Gis group and 22 points in the control group (Table 3). The effect persisted in the AT S100 + AT TNF-α + AT His group after 6 h of observation: 24 vs 18 points in the AT His group and control. After 24 hours, there was a decrease in excretory activity in all three experimental groups until the end of defecation in 45% of the animals.

Thus, it was shown that the complex preparation AT S100 + AT TNF-α + AT Gis has a mild laxative effect, superior in effectiveness to AT gis.

Example 3

In the study of the antispasmodic activity of the claimed complex drug, 30 outbred male mice (weight 17.5-26.3 g, age 1.5-2 months) were used, which were administered intragastrically for 5 days with distilled water (control, 15 ml / kg), AT His (15 ml / kg) or AT S100 + AT TNF-α + AT His (15 ml / kg). The antispasmodic activity of the preparations was evaluated according to the method of J. Setnicar (Setnicar J., Da Re P. 3-methyl-6 (N-diethyl-amino-methyl) -Flavone - a new smooth-muscle relaxant // Arzneimittel-Forsch. 1959. No .9. P.653-697).

One hour after the last administration of the preparations, mice were injected intraperitoneally with 0.2 ml of a 0.1% BaCl ₂ solution and intragastrically with 0.5 ml of a 10% suspension of activated charcoal prepared in 2% potato starch mucus. After 10 minutes, the animals were sacrificed and the ratio between the total length of the intestine and the charcoal filled was determined. The ratio was expressed as a percentage and was taken into account as the primary result of the experiment.

In the group of mice treated with the AT S100 + AT TNF + AT Gis preparation, the maximum distance of coal advancement in the intestines of mice was observed: the length of the ulcer-filled part was 1.3 times longer than the similar parameters of the AT Gis group and control (Table 3). That is, a gastrointestinal spasm caused by the introduction of BaCl ₂ and leading to a decrease in the rate of advancement of activated carbon through the intestines was least expressed in the AT S100 + AT TNF-α + AT His group.

Table 3 Evaluation of the antispasmodic effect of the tested drugs by the method of "carbon labels" (with BaCl ₂ ) Monitoring group (number of animals) The length of the part of the intestine filled with coal per 1 mouse (M ± m), cm The length of the coal-filled part to the total length of the intestine (M ± m),% Control (n = 10) 23.3 ± 2.20 42.4 ± 3.49 AT Gis (n = 10) 22.3 ± 2.09 40.1 ± 3.81 AT S100 + AT TNF-α + ATGis (n = 10) 30.0 ± 2.42 * # 56.4 ± 4.50 * # * - differences are significant relative to the control group (p <0.05) # - differences are significant relative to the AT Gis group (p <0.05) Thus, it was shown that the complex preparation AT S100 + AT TNF-α + AT Gis has a moderate antispasmodic effect, superior in effectiveness to AT Gis.

Thus, it was shown that the complex preparation AT S100 + AT TNF + AT Gis has a moderate antispasmodic effect, superior in effectiveness to AT Gis.

Example 4

In the study of the properties of the claimed drug for the treatment of patients of the active drug group, 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 human tumor necrosis factor alpha (AT TNF-α), brain-specific protein S-100 (AT S100) and histamine (AT His) in ultra-low doses (SMD) obtained by super-dilution of the initial concentration 2 matrix solution, 5 mg / ml 100 ¹² , 100 ³⁰ , 100 ²⁰⁰⁰ times, equivalent to a mixture of hundreds of homeopathic dilutions of C12, C30, C200. To treat patients of the comparison group, 300 mg tablets were used, impregnated with an aqueous-alcoholic solution (3 mg / tablet) of an activated, potentiated form of polyclonal rabbit antibodies to histamine, purified on an antigen, in an ultra-low dose (SMD AT Gis) obtained by super-dilution of the initial a matrix solution of a concentration of 2.5 mg / ml in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, the equivalent mixture of hundred-percent homeopathic dilutions of C12, C30, C200 (comparison drug).

A comparative study evaluating the effectiveness of the composition of SMD AT TNF-α + AT S100 + AT Gis and the monocomponent drug AT Gis in the treatment of patients with irritable bowel syndrome (IBS) involved 52 patients with a diagnosis verified in accordance with Rome III (2006), which underwent an outpatient course of observation and therapy for 12 weeks. 24 study participants were included in the active drug group (DMF AT TNF-α + AT S100 + AT Gis 2 tablets 2 times a day), 28 - in the comparison drug group (SMD AT GIS 2 tablets 2 times a day). Both groups of patients were comparable in terms of baseline demographic, anthropometric, and clinical laboratory parameters. 18 patients had IBS with constipation (hard or lumpy stools accounted for more than 25%, and loose stools - less than 25% of all bowel movements), 14 - IBS with diarrhea (mushy or loose stools made up more than 25%, and hard stools - less than 25% all bowel movements) and 20 - a mixed variant of IBS (respectively, hard, lumpy stools and loose stools accounted for more than 25% of all bowel movements). As criteria for the effectiveness of therapy, a decrease in the intensity of pain / discomfort (weekly average, fluctuations from 0 to 10 points) compared with the initial state, as well as the dynamics of other dyspeptic symptoms on the VAS-IBS (Visual Analogue Scale-Irritable Bowel Syndrome ( VAS-IBS): Guidance for Industry Irritable Bowel Syndrome - Clinical Evaluation of Products for Treatment. US Department of Health and Human Services; Food and Drug Administration; Center for Drug Evaluation and Research (CDER). - March 2010; Muller-Lissner, S, G Koch, NJ Talley et al., 2004, Subject's Global Assessment of Relief: An Appropriate Method to Assess the Impact of Treatment on IBS-Related Symptoms in Clinical Trials, J Clin Epidemiol, 56: 310-316; CPMP / EWP / 785/97, 2003, Points to Consider on the Evaluation of Medicinal Products for 483 the Treatment of Irritable Bowel Syndrome, London, Available at: 484 http://www.emea.europa.eu/pdfs/human/ewp/078597en.pdf); Visceral Sensitivity Index (VSI): Labus S. Jennifer et al. The Central Role of Gastrointestinal-Specific Anxiety in Irritable Bowel Syndrome: Further Validation of the Visceral Sensitivity Index Psychosomatic Medicine, 2007; 69: 89-98) and HADS score (Hospital Anxiety And Depression Scale (HADS): Snaith R. Philip. The Hospital Anxiety And Depression Scale. Health and Quality of Life Outcomes 2003, 1: 1-4. Http://www.hqlo.com/content/1/1/29); in patients with IBS with a predominance of diarrhea, the proportion of patients with a change in stool type was calculated according to the Bristol Stool Shape Scale (Guidance for Industry Irritable Bowel Syndrome - Clinical Evaluation of Products for Treatment. US Department of Health and Human Services; Food and Drug Administration; Center for Drug Evaluation and Research (CDER). - March 2010.) up to 5 or less (but not lower than ≤2 on average per week); in patients in the IBS subgroup with a predominance of constipation, the percentage of patients with an increase in the number of bowel movements on average 1 time per week compared to the original.

In the clinical picture of the disease, pain abdominal syndrome prevailed in both groups (average score 7.56 ± 0.26 in the active drug group and 7.21 ± 0.24 in the VAS-IBS comparison group) (Table 4), which occurred in 100% of patients at the initial examination. Among dyspeptic manifestations, diarrhea (in 54% of patients of the active drug group and in 57% of patients in the comparison group) and constipation (62% and 57%, respectively), bloating and flatulence (in 46% and 36%, respectively) were recorded equally, the severity of which was approximately the same in two groups (Table 4). Nausea and vomiting were less common (30% and 32%, respectively). The average visceral sensitivity index (VSI) and HADS score did not differ in both groups (Table 4), indicating a significant effect of disturbances on the part of the central nervous system in the development of IBS. In addition to the studied therapy, patients of both groups were allowed to take the laxative Guttalax® and the antidiarrheal Smecta® for relieving constipation or diarrhea, respectively, as well as No-shpa® to reduce the severity of spastic abdominal pain. These drugs were taken by patients as needed. All patients of the study groups completed the treatment within the time period established by the study protocol; there were no prematurely retired patients.

Table 4 The dynamics of the main indicators depending on the type of therapy Period AT TNF-α + AT S100 + AT His (n = 24; M ± SE) AT Gis (n = 28; M ± SE) VAS-IBS: pain / discomfort, points Originally 7.56 + 0.26 7.21 ± 0.24 12 weeks 3.32 ± 0.13 *** 5.64 ± 0.24 ** VAS-IBS: diarrhea, points Originally 5.52 ± 0.18 5.73 ± 0.34 12 weeks 3.34 ± 0.22 * 4.86 ± 0.22 VAS-IBS: constipation, points Originally 5.79 ± 0.26 4.98 ± 0.34 12 weeks 4.41 ± 0.33 ** 4.25 + 0.26 VAS-IBS: bloating and flatulence, points Originally 4.98 ± 0.34 4.57 + 0.31 12 weeks 3.84 ± 0.24 ** 4.04 ± 0.31 VAS-IBS: vomiting and nausea, points Originally 3.94 + 0.31 3.56 ± 0.24 12 weeks 2.87 ± 0.12 * 3.03 ± 0.27 Visceral Sensitivity Index (VSI-IBS), points Originally 22.3 ± 1.6 25.4 ± 1.8 12 weeks 68.7 ± 2.4 *** 33.9 ± 1.7 HADS Hospital Anxiety and Depression Scale, points Originally 19.3 ± 1.4 18.9 ± 1.5 12 weeks 12.8 ± 0.6 *** 14.4 ± 1.3 Note. * significance of differences between groups AT TNF-α + AT S100 + AT Gis and AT Gis, p <0.05; ** significance of differences with the initial indicator, p <0.05.

The analysis of efficacy revealed that during the 12-week therapy the severity of the main clinical manifestation of IBS - abdominal pain syndrome - in patients of the group receiving the composition of SMD AT TNF-α + AT S100 + AT Gis decreased on average by more than 50% compared with the initial state (up to 3.32 ± 0.13 points). This decrease had significant differences when compared with the results of treatment with the reference drug (SMD AT Gis), which also helped to reduce the severity of pain / discomfort, but this decrease was less significant (less than 30%).

Acceptance of the composition of SMD AT TNF-α + AT S100 + AT Gis had a positive effect on other dyspeptic disorders in patients, including diarrhea (reduction of the initial 5.52 ± 0.18 points to 3.34 ± 0.22 points by the end of therapy), constipation (5.79 ± 0.26 and 4.41 ± 0.33 points, respectively), bloating and flatulence (4.98 ± 0.34 and 3.84 ± 0.24 points, respectively), and the effectiveness in relation to the last two the symptoms had significant differences when compared with both the initial state and the effectiveness of therapy with the monocomponent drug SMD AT Gis.

In the subgroup of patients with IBS with a predominance of diarrhea (n = 14), the proportion of patients with a change in stool type on the Bristol scale of stool shape to 5 or less as a result of a 12-week course of therapy with SMD AT TNF-α + AT S100 + AT Gis was 57% ; in the IBS subgroup with a predominance of constipation (n = 18), the percentage of patients with an increase in the number of bowel movements on average 1 time per week reached 94%; among patients with a mixed IBS variant (n = 20), similar indicators were 45% and 75%, respectively. Among patients in the comparison group, the studied indicators did not have significant dynamics.

The effectiveness of treatment with the SMD AT TNF-α + AT S100 + AT Gis composition was manifested in a positive effect on visceral hypersensitivity, which significantly decreased after 12 weeks of taking the combined drug, as a result of which the VSI index increased from 22.3 ± 1.6 to 68, 7 ± 2.4 (versus 25.4 ± 1.8 and 33.9 ± 1.7, respectively, in the comparison drug group). Positive shifts under the influence of the composition of SMD AT TNF-α + AT S100 + AT GIS also manifested themselves in a decrease in the total score on the HADS scale (from 19.3 ± 1.4 to 12.8 ± 0.6), which indicated the relief of the initial a place of subclinical anxiety and depression.

The safety assessment of therapy, which was carried out on the basis of registration of adverse events during treatment and re-examination of laboratory parameters, indicated good tolerance to the drugs. The safety analysis included data from all patients participating in the study (n = 52). During the 12-week course of treatment, not a single patient from both groups revealed a single adverse event that had a “possible” or “obvious” connection with taking the drug. Laboratory studies, including general and biochemical blood tests, a clinical analysis of urine, also did not record any pathological abnormalities during therapy.

Thus, the study demonstrated the effectiveness and safety of the composition of SMD AT TNF-α + AT S100 + AT His in the treatment of patients with IBS. It was shown that a 12-week intake of the composition of SMD AT TNF-α + AT S100 + AT Gis contributed to the relief of abdominal pain syndrome, as well as a significant decrease in the severity of dyspeptic manifestations in patients with IBS. The treatment effect was confirmed by a high percentage of patients who improved bowel movements. In addition to the positive dynamics of the main symptoms of the gastrointestinal tract, a normalization of the patient's somatic and mental state was observed, which was expressed in positive changes in visceral sensitivity and the leveling of symptoms of anxiety and depression. All the noted effects of therapy with the SMD AT TNF-α + AT S100 + AT Gis composition had significant differences compared with the initial state and the results of treatment with the SMD AT Gis. The safety profile was equally high for both drugs. Both the composition of SMD AT TNF-α + AT S100 + AT Gis and the monocomponent drug SMD AT Gis did not lead to the appearance of adverse events or pathological deviations of laboratory parameters associated with the administration of the studied drugs.

Claims (11)

1. A drug for the treatment of functional bowel disorders, containing an activated-potentiated form of antibodies to histamine, characterized in that it is made in the form of a pharmaceutical composition and contains, as additional - enhancing components, an activated-potentiated form of antibodies to tumor necrosis factor-alpha (TNF- α) and the activated-potentiated form of antibodies to the brain-specific protein S-100. 2. The drug according to claim 1, characterized in that the activated-potentiated form of antibodies to histamine, the activated-potentiated form of antibodies to TNF-α and the activated-potentiated form of antibodies to the brain-specific protein S-100 are used in the form of activated-potentiated aqueous or aqueous-alcoholic solution obtained in the process of successive multiple dilutions of the matrix solution of the corresponding antibodies in an aqueous or aqueous-alcoholic solvent and an intermediate external mechanical zdeystviya - shaking. 3. The 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 an activated-potentiated form of antibodies to histamine, an activated-potentiated form of antibodies to TNF-α and an activated-potentiated form of antibodies to the brain-specific protein S-100; and pharmaceutically acceptable additives. 4. The drug according to claim 1 or 2, characterized in that aqueous or aqueous-alcoholic solutions of activated-potentiated forms of antibodies to histamine, to TNF-α and to brain-specific protein S-100 are obtained by repeated serial dilution and intermediate external exposure from matrix solutions of affinity-purified antibodies to histamine, to TNF-α and to the brain-specific protein S-100 with a concentration of 0.5 ÷ 5.0 mg / ml. 5. The drug according to claim 1 or 2, characterized in that each of the components of the smallest doses of affinity-purified antibodies is used as a mixture of various, mainly hundred, homeopathic dilutions. 6. The drug according to claim 3, characterized in that the pharmaceutically acceptable additives include lactose, microcrystalline cellulose and magnesium stearate. 7. A method of treating functional intestinal disorders by introducing into the body a drug containing an activated-potentiated form of anti-histamine antibodies, characterized in that it additionally simultaneously combines the activated-potentiated form of ultra-low doses of affinity-purified antibodies to TNF-α and the activated-potentized form antibodies to the brain-specific protein S-100. 8. The treatment method according to claim 7, characterized in that a mixture of different homeopathic dilutions of antibodies to histamine combined with a mixture of different homeopathic dilutions of antibodies to TNF-α and a mixture of different homeopathic dilutions of antibodies to brain-specific protein S-100. 9. The method according to claim 7, characterized in that said functional bowel disorder is irritable bowel syndrome. 10. The method according to claim 7, characterized in that said functional bowel disorder is a violation of the motor-evacuation function of the intestine. 11. The method according to claim 7, characterized in that the activated-potentiated form of antibodies to histamine, the activated-potentiated form of antibodies to TNF-α and the activated-potentiated form of antibodies to the brain-specific protein S-100 are used in the form of an activated-potentiated aqueous or aqueous -alcohol solution obtained in the process of sequential multiple dilution in an aqueous or aqueous-alcoholic solvent and an intermediate external mechanical action - shaking.