UA112751C2 - Combination pharmaceutical composition and methods of treating diseases or conditions associated with neurodegenerative diseases - Google Patents

Abstract

The present invention relates to combination pharmaceutical composition comprising an activated-potentiated form of an antibody to gamma interferon, and an activated-potentiated form of an antibody to S-100 protein and method of treating multiple sclerosis and other neurodegenerative diseases, as well as the diseases and conditions associated with neuroinfections.

Description

(57) Abstract:

The invention relates to a combined medicinal product containing a mixture of solutions of homeopathically potentiated forms of a) antibodies to gamma interferon in a mixture of three homeopathic dilutions C12, C30 and C 200 or C12, C30 and C50, and B) antibodies to protein 5-100 in a mixture of three homeopathic dilutions C12, C30 and C 200 or C12, C30 and C50. The invention also relates to methods of treating or reducing the frequency of multiple sclerosis relapses, treating other neurodegenerative diseases and diseases and conditions associated with neuroinfections.

TECHNICAL FIELD

The present invention relates to combined medicinal preparations and methods of treatment of multiple sclerosis and other neurodegenerative diseases, as well as diseases or conditions associated with neuroinfections.

TECHNICAL LEVEL

Multiple sclerosis (MS) is a chronic disease that affects the brain and spinal cord, resulting in loss of muscle control, balance, and sensation (such as numbness). At this time, the exact cause of MS remains unknown, but researchers believe that a combination of factors may contribute. It is believed that MS occurs in genetically predisposed individuals, probably in the presence of specific external factors that determine the development

RUS It has now been established that an autoimmune process is involved in MS - an abnormal reaction of the body's immune system directed against myelin (the fatty sheath that surrounds and envelops nerve fibers) in the central nervous system (CNS - the brain, spinal cord and optic nerves).

MS leads to thinning or complete loss of myelin and, as the disease progresses, cutting (severing) of neuron processes (axons). With the loss of myelin, the neuron is unable to conduct electrical signals effectively. A repair process called "remyelination" occurs in the early stages of the disease, but the myelin sheath of the cell cannot be completely restored. Repeated attacks lead to fewer instances of successful and efficient remyelination until a scar-like plaque forms around the damaged axons.

In addition to myelin loss, another pathological feature of the disease is inflammation.

According to a purely immunological explanation, the inflammatory process is caused by T cells, a type of lymphocyte. Lymphocytes are cells that play an important role in the protective functions of the body. With ROS, T-cells are able to penetrate the brain through the hemoencephalitic barrier.

T cells are thought to perceive myelin as foreign and attack it, leading to inflammation, stimulation of other immune cells, and soluble factors such as cytokines and antibodies.

In addition to an autoimmune disorder, some researchers believe that infections can

Koo) somehow "force" the immune system to attack nerve cells. In general, the virus (or bacteria) causing the original infection is thought to "look" like a nerve cell. The immune system produces T-cells to fight the virus. These T cells remain in the body after the infection has been eliminated, and after "seeing" the nerve cell, they mistake it for the pathogen. As a result, the immune system attacks the nervous system.

There are four main varieties of ROS. 1. Relapsing/relapsing multiple sclerosis (MS): Characterized by relapses in which new symptoms may appear and old ones may reappear or worsen. 2. Secondary progressive multiple sclerosis (PSMS): characterized by gradual worsening of the disease in the periods between relapses. 3. Progressive relapsing multiple sclerosis (RPMS): This form of MS is progressive from the onset of the disease and is punctuated by relapses. 4. Primary progressive multiple sclerosis (PPMS), this type of MS is characterized by a gradual progression of the disease from the very beginning without remissions at all.

Currently, there is no known cure for MS. However, there are types of therapy that can slow down the disease. The goal of treatment is to control symptoms. Medicines that correct the immune system, such as interferons, are used to treat multiple sclerosis. Interferons are protein messengers produced by cells of the immune system and used by them to communicate with each other. There are different types of interferons, such as alpha, beta and gamma. All interferons are able to regulate the immune system and play an important role in defense against foreign organisms, including viruses. All interferons have different functions, which at the same time overlap. Beta interferons have been found to be useful in controlling multiple sclerosis.

There is a constant need to find new drugs that have the desired therapeutic efficacy in the treatment of MS and related symptoms.

The therapeutic effect of the maximally diluted (or ultralow) form of antibodies potentiated with the help of homeopathic technology (activated potentiated form) was discovered by the inventor of this patent application, Dr. O.I. Epstein. US Patent No. 7,582,294 describes an agent for the treatment of benign prostatic hyperplasia or prostatitis by administering a homeopathically activated form of antibodies to prostate-specific antigen (PSA). because Protein 5-100 is a cytoplasmic acidic calcium-binding protein that is formed mainly in the gray matter of the brain, primarily in glia and Schwann cells.

The protein exists in several homo- or heterodimeric isoforms, consisting of two immunologically separated subunits - alpha and beta. Protein 5-100 is proposed for use as an aid in the diagnosis and evaluation of brain lesions and neurological damage resulting from traumatic brain injury such as stroke. See Magdap ei a!I., OveitsIpev5 oi 51008 Rgoivip ip Meshgoiodisa! Oizogaetgtv5, ) Cancer Mei Avzos Moi. 61, Mo. 3, Magsy 2011, which is incorporated into this document by reference.

It was established that ultra-low doses of antibodies to protein 5-100 have anxiolytic, anti-asthenic, anti-aggressive, stress-protective, anti-hypoxic, anti-ischemic, neuroprotective and nootropic activity. See Savziadpe M. ei aiYi., Apiirodieye tyu 5100 rgoivipe Name aphioiuis-iike asimpu ai iPga-Iom/ dose5 ip adiy gai, In Rpapt RNaptasoi. 2008, 60(3):309-16; Epstein 0. 1.,

Antibodies to calcium-binding protein 51008 block the formation of long-term sensitization in the land snail, Rnaptasoi! Viosnet Vepam., 2009, 94(1):37-42; Voronina T.A. et al., Chapter 8. Antibodies to protein 5-100 in anxiety-depressive disorders in experimental and clinical conditions. In the book "Animal models in biological psychiatry", ed.

Kaluyeva A. V. M-U, "Moma zZsiepse Rybiizpegv, Ips.", 2006, p. 137-152, which are incorporated herein by reference.

It was found that ultra-low doses of antibodies to gamma interferon are useful in the treatment and prevention of diseases of viral etiology. See US Patent No. 7,572,441, which is hereby incorporated by reference in its entirety.

BRIEF DESCRIPTION OF THE INVENTION

In one aspect, the present invention provides a combined medicinal product containing a) an activated potentiated form of an antibody to gamma interferon and b) an activated potentiated form of an antibody to protein 5-100.

In one variant, the present invention is a combined medicinal product containing a) an activated potentiated form of an antibody to gamma-interferon and b) an activated potentiated form of an antibody to protein 5-100, where an antibody to the whole gamma-interferon or its parts is present.

In one embodiment, the present invention is a combined medicinal product containing a) an activated potentiated form of an antibody to gamma interferon and b) an activated potentiated form of an antibody to protein 5-100, where the antibody to protein 5-100 is an antibody to

From the whole protein 5-100 or its parts.

In one embodiment, the combined medicinal product of this aspect of the invention includes an activated potentiated form of the gamma-interferon antibody in the form of a mixture of homeopathic dilutions (C12, C30, and C50) or (C12, C30, and C200) impregnated on a solid carrier. The activated potentiated form of antibody to protein 5-100, presented in the form of a mixture of homeopathic dilutions (C12, C30, and C50) or (C12, C30 and C200), can subsequently be impregnated on a solid support.

In another embodiment, the combined medicinal product of this aspect of the invention includes an activated potentiated form of antibody to protein 5-100 in the form of a mixture of homeopathic dilutions (C12, C30, and C50) or (C12, C30, and C200) impregnated on a solid carrier. The activated potentiated form of the gamma-interferon antibody, presented in the form of a mixture of homeopathic dilutions (C12, C30, and C50) or (C12, C30 and C200), can subsequently be impregnated on a solid support.

If possible, the activated potentiated form of the antibody to gamma-interferon is a monoclonal, polyclonal or natural antibody, preferably a polyclonal antibody. In one embodiment of this aspect of the invention, the activated potentiated form of the gamma-interferon antibody is prepared by serial hundredfold dilutions in combination with shaking of each dilution.

If possible, the activated potentiated form of antibody to protein 955-100 is a monoclonal, polyclonal or natural antibody, preferably a polyclonal antibody. In one embodiment of this aspect of the invention, the activated potentiated form of the antibody to protein 5-100 is prepared by successive hundredfold dilutions in combination with shaking of each dilution.

Vertical shaking is specially provided.

In another aspect, the invention provides a method of treating patients suffering from multiple sclerosis by administering a) an activated potentiated form of an antibody to gamma interferon and b) an activated potentiated form of an antibody to protein 595-100. If possible, the activated potentiated form of anti-gamma-interferon antibody and the activated potentiated form of anti-protein 5-100 antibody should be administered in the form of a combined medicinal product. In another aspect, the invention provides a method of significantly delaying the onset of symptoms in patients suffering from multiple sclerosis by administering a combined medicinal product, wherein the medicinal product contains a) an activated potentiated form of an antibody to gamma interferon and B) an activated potentiated form of an antibody to protein 5 -100.

In another aspect, the present invention also provides a method of reducing the frequency of relapses in patients suffering from multiple sclerosis by administering a combination medicinal product, wherein the medicinal product contains a) an activated potentiated form of an antibody to gamma interferon, and B) an activated potentiated form of an antibody to protein 5-100.

In one variant of the invention, it is envisaged to introduce from 1 to 2 standard dosage forms of the activated potentiated form of the antibody to gamma-interferon, and from 1 to 2 standard dosage forms of the activated potentiated form of the antibody to protein 5-100, and each dosage form is administered from 1 to 4 times day. If possible, 1-2 standard dosage forms of each activated potentiated form of antibody should be administered twice daily.

A preferred embodiment of this aspect of the invention provides for the administration of 1 to 2 standard dosage forms of a combined preparation containing a) an activated potentiated form of an antibody to gamma interferon and B) an activated potentiated form of an antibody to protein 5-100, and each dosage form the form is administered from 1 to 4 times a day.

If possible, 1-2 standard dosage forms of each activated potentiated form of antibody should be administered twice daily.

In another preferred embodiment of this aspect of the invention, the combination drug is administered as 1 standard dosage form containing a) an activated potentiated form of the anti-gamma interferon antibody and B) an activated potentiated form of the anti-protein 5-100 antibody, preferably twice day.

DESCRIPTION OF DRAWINGS

Fig. 1 - illustrates the immune reaction in the pathogenesis of multiple sclerosis

Fig. 2 - shows the average period of onset of clinical symptoms

Fig. C - shows the severity of the disease (points)

Fig. 4 - shows the degree of severity at different stages (points)

Fig. 5 - shows the frequency of relapses

DETAILED DESCRIPTION OF THE INVENTION

The invention is defined with reference to the appended claims. For statements, the following glossary provides relevant definitions.

The term "antibody" as used herein means an immunoglobulin that specifically binds (and is accordingly defined as complementary) to a specific spatial and polar organization of another molecule. As indicated in the claims, antibodies may include all or part of an immunoglobulin, may be natural, polyclonal or monoclonal, and may include different classes and isotypes such as IDA, dO, I9E, Ide, Idoga,

IChS2b and IdS3, I9M, etc. Its parts may include Rab, Em and E(ar)2, Bar', etc.

The term "antibody" in the singular includes "antibodies" in the plural. The term "activated potentiated form" or "potentiated form", respectively, in relation to the antibodies specified herein, is used to refer to the product of homeopathic potentiation of any original antibody solution. "Homeopathic potentiation" means the application of homeopathic methods to add homeopathic activity to a stock solution of a corresponding substance.

Although the term is not so limited, "homeopathic potentiation" may include, for example, repeated serial dilutions in combination with external treatment, in particular, vertical (mechanical) shaking. In other words, the original antibody solution is subjected to serial repeated dilutions and repeated vertical shaking of each resulting solution according to homeopathic technology. The preferred concentration of the original antibody solution in the solvent, preferably water or a mixture of water and ethyl alcohol, varies from about 0.5 to 5.0 mg/ml. The preparation procedure of each component (ie, the antibody solution) to which the preference is to use a mixture of C aqueous or aqueous-alcoholic dilutions of the primary matrix solution (mother tincture), antibodies diluted 10072, 10030, and 100200 times, respectively, which is equivalent to hundredth homeopathic dilutions (C12, C30, and C200), or using mixture of three water or water-alcohol dilutions of a well matrix solution of antibodies diluted 10072, 10030 and 10059 times, respectively, which is equivalent to hundred homeopathic dilutions (C12, C30 and C50). Examples of homeopathic potentiation are described in US Pat

Nos. 7,572,441 and 7,582,294, which are hereby incorporated by reference in their entirety for the stated purpose. While the term "activated potentiated form" is used in the claims, the examples use the term "ultra-low doses". The expression "ultra-low doses" became a special term created during the research and use of homeopathically diluted and potentized form of the substance. The term "ultra-low dose" or "ultra-low doses" fully supports and is primarily synonymous with the term "activated potentiated" form used in the claims.

In other words, the antibody is in an "activated potentiated" or "potentiated" form in the presence of three factors. First of all, the "activated potentiated" form of the antibody is the product of a preparation process well known in the homeopathic field. Secondly, the "activated potentiated" form of the antibody must have biological activity determined by methods accepted in modern pharmacology. Thirdly, the biological activity manifested by the "activated-potentiated" form of the antibody cannot be explained by the presence of the molecular form of the antibody in the final product of the homeopathic process.

For example, an activated potentiated form of antibodies can be prepared by subjecting the original, isolated antibody in molecular form to successive multiple dilutions in combination with an external action, for example, mechanical shaking. External processing in the course of reducing concentration can also be accompanied, for example, by the influence of ultrasonic, electromagnetic or other physical factors. W. Schwabe "Homeopathic Medicines", M., 1967, US Patents MoMo 7,229,648 and 4,311,897, which are fully incorporated herein by reference and for the stated purpose, describe such processes which are common methods of homeopathic potentiation in homeopathic practice.

This procedure causes a uniform decrease in the molecular concentration of the initial molecular form of the antibody. This procedure is repeated until the required homeopathic activity is obtained. For an individual antibody, the required homeopathic activity can be determined by subjecting intermediate dilutions to biological tests in the desired pharmacological model. Although the term is not so limited, "homeopathic potentiation" may include, for example, repeated serial dilutions combined with external treatments, particularly vertical (mechanical) shaking. In other words, the initial antibody solution is subjected to successive repeated dilutions and repeated vertical shaking of each obtained solution according to homeopathic technology.

The preferred concentration of the original solution of the antibody in the solvent, preferably water or a mixture of water and ethyl alcohol, varies from about 0.5 to 5.0

From mg/ml. A preferred procedure for the preparation of each component (i.e., the antibody solution) is to use a mixture of 3 aqueous or aqueous-alcoholic dilutions of the primary matrix solution (mother tincture) of the antibodies diluted 10072, 10030, and 100200, respectively, which is equivalent to one-hundredth homeopathic dilution C12, C30 and C200, or the use of a mixture of 3 aqueous or water-alcohol dilutions of the primary matrix solution (mother tincture) of antibodies diluted 10072, 10030 and 10059 times, respectively, which is equivalent to hundreds of homeopathic dilutions of C12, C30 and C50. Examples of obtaining the desired activity are also provided, for example, in US Patents MoMo 7,229,648 and 4,311,897, which are incorporated herein by reference for the stated purpose. The procedure applicable to the "activated potentiated" form of the antibody described herein is described in more detail below.

There is a lot of controversy in scientific circles about homeopathic treatment of people. While this invention is based on generally accepted homeopathic processes for obtaining an "activated potentiated" form of antibodies, it is not only based on the use of homeopathy in humans to prove its effectiveness. The inventor unexpectedly discovered and, in this application to a sufficient extent, demonstrated in conventional pharmacological models that the solvent ultimately obtained after successive multiple dilutions of the original molecular form of antibodies had significant activity unrelated to the presence of residues of the molecular form of the antibody in the target dilution. An "activated potentiated" form of an antibody as defined herein is tested for biological activity in conventional pharmacological activity models, or in appropriate ip miigo or ip mimo experiments in suitable animal models. The experiments described below confirm the biological activity in such models. Human clinical studies also confirm that the activity observed in animal models correlates well with treatment in humans. Studies in humans have also confirmed the existence of the "activated-potentiated" forms described herein for the treatment of certain diseases or disorders in humans generally accepted in medicine as pathological conditions.

In addition, the declared "activated potentiated" form of the antibody covers only solutions or solid preparations whose biological activity cannot be explained by the presence of the molecular form of the antibody remaining from the initial, initial solution. In other words, although it is assumed that the "activated potentiated" form of the antibody may contain remnants of the original molecular form of the antibody, one skilled in the art cannot attribute the biological activity observed in known pharmacological models to the remaining molecular form of the antibody with at least some degree of probability, given very low concentrations of the molecular form of the antibody remaining after serial dilutions. While the invention is not limited by any particular theory, the biological activity of the "activated-potentiated" form of the antibodies of the present invention is not related to the original molecular form of the antibody. Preference is given to the "activated-potentiated" form of the antibody in the liquid or solid state, in which the concentration of the original molecular form of the antibody is below the threshold of detection according to conventional analytical methods such as capillary electrophoresis and high performance liquid chromatography. In particular, an "activated potentiated" form in the liquid or solid state in which the concentration of the original molecular form of the antibody is below Avogadro's number is preferred. In the pharmacology of molecular forms of therapeutic substances, it is common practice to draw a dose-response curve, in which the level of pharmacological response is plotted against the concentration of the active drug administered to the patient or tested i.p. The minimum level of the drug that provides at least some the detectable response is called the threshold dose. It is hereby specifically contemplated and preferred that the "activated potentiated" antibody form contains the molecular antibody, if present, at a concentration below the threshold dose for the molecular form of the antibody in a given biological model.

Known experimental models of multiple sclerosis. For example, Experimental Autoimmune Encephalomyelitis, sometimes referred to as Experimental Allergic Encephalomyelitis (EAE), is an animal model of inflammatory demyelinating diseases of the central nervous system. It is mostly used on rodents and knockout mice. This experimental model is a widely studied animal model of demyelinating diseases of the human central nervous system, such as multiple sclerosis. EAE can be induced in susceptible animals with the help of a homogenate of homologous or heterologous brain tissue, purified myelin or proteins introduced into the preparation (Maipe, ei aiYi., App. MU Asad. 5si., 1984, 436: 33-51; MeSotre, evy ai., U. Meishgoittipo!). 1994; 51 (2): 153-167).

At the moment, more than 20 myelin proteins have been described that have immunogenic properties and are most often used to induce EAE (Vaitapp, ei ai., Rpuzioi. Chem.

Zo 2001, 81(2): 871-927). Among them: basic myelin protein ("MVR") (Nazpyit, Ittipoi. Chem., 1978, 39:60-107); proteolipid protein ("RI P") (Vgadi, eyi ai., Vgaip Raiyoi!., 1996, 6(3): 303-311;

Tyopu, Meshhospet. Nez., 1994, 19(8):935-944); glycoproteins: myelin-associated glycoprotein (SMAS") (Mueegii ei aiYi, 1999); myelin oligodendrocyte glycoprotein (MO); and oligodendrocyte-specific protein (O5P") (5iemepv, ei ai., U Ittipoi., 1999; 162 (12 ): 7501-1509). It is generally accepted that not only whole proteins, but also specific parts of these proteins are capable of causing EAE when administered to animals. The most commonly used antigens in rodents are spinal cord homogenate (SHC), purified myelin, myelin protein such as MVR, RI P, and

MOSS or peptides of these proteins, each of which drives distinct patterns with different disease characteristics in relation to both immunology and pathology.

The most representative model of MS is induced in EAE models by the introduction of homogenate of the spinal cord (Hilerovych et al., 2010; Zhabotinsky, Joffe, 1975;

Zhitnukhin et al., 2008; 5ippa ei ai, 2009). In this model, as in the original disease, an immune reaction to all components of myelin is provided: it is caused with subsequent demyelination, disintegration of axons and then the nerve cells themselves (Hilerovych et al., 2010;

Zippa Yeyi, 2009). The pathological chain of events manifests itself through the development of paresis, paralysis and other symptoms of the disease.

Four stages of the development of EAE can be established: 1. Sensitization of peripheral T-lymphocytes under the influence of cerebral antigen from CEA and increased permeability of SEV. 2. Migration of activated T cells to the CNS and activation of antigen-representing cells (astrocytes and microglia) directly in the brain. These two stages correspond to the latent period (induction), when clinical manifestations are not observed 3. Development of autoimmune and inflammatory reactions in the brain, which leads to demyelination of nerve fibers (clinical period). 4. Suppression of pathological processes and restoration of damaged tissues (recovery phase). During this period, most animals resolve neurological and motor disorders, after which complete or partial recovery occurs and animals become resistant to repeated EAE stimulation. The average duration of EAE is 30 days: latent stage, clinical stage and recovery stage. The duration of each stage, as a rule, is 7-10 days. because Analogous phases of the development of the pathological process in the central nervous system are also observed in multiple sclerosis. See Fig. 1 - Immune reaction in the pathogenesis of multiple sclerosis (Umiepai 5

Kieseig, 2003).

The present invention describes a combined drug containing a) an activated potentiated form of an antibody to gamma interferon and B) an activated potentiated form of an antibody to protein 5-100. As stated above in this document, each of the individual components of the combination is well known and widely used in medical practice. However, the inventors of this application were surprised to find that the introduction of the combination significantly delays the onset of symptoms and reduces the likelihood of relapse in patients with multiple sclerosis.

The combined medicinal product according to this aspect of the invention may be in solid or liquid form. Each of the activated-potentiated forms of antibodies contained in the drug is prepared from the original molecular form of antibodies using a process accepted in homeopathic practice. The starting antibodies can be monoclonal or polyclonal antibodies, prepared according to known processes, for example, as described in the literature: Immunological methods, H. Frimel, M., "Medicine", 1987, p. 9-33; "Thread

Apiirodiev. Moposiopai! apa gesotbipapi apiibodiez 30 uyeagz apeg!", anu E., Zodoueg V. - 2005 - Moi. 14. - M 1-2. P.33-55, which are included in this document by reference.

Monoclonal antibodies can be obtained, for example, using hybridoma technology.

The initial stage of the process involves immunization based on principles previously developed during the preparation of polyclonal antiserum. The next stages of work include the production of hybrid cells that generate clones of antibodies with identical specificity. Their separate isolation is carried out using the same methods as in the preparation of polyclonal antisera.

Polyclonal antibodies can be obtained by active immunization of animals. For this, suitable animals (for example, rabbits) are given a series of injections of the appropriate antigen: protein 5-100 or gamma interferon. The immune system of animals produces appropriate antibodies that are collected in animals in a known manner. This procedure makes it possible to prepare a monospecific serum saturated with antibodies.

If desired, serum containing antibodies can be purified, for example, by affinity chromatography, fractionation by salt precipitation, or ion exchange chromatography. The resulting purified, antibody-saturated serum can be used as a starting material for the preparation of an activated potentiated form of antibodies.

The preferred concentration of the obtained initial solution of the antibody in the solvent, preferably water or a mixture of water and ethyl alcohol, varies from about 0.5 to 5.0 mg/ml.

A preferred procedure for the preparation of each component is to use a mixture of three aqueous-alcoholic dilutions of the primary antibody matrix diluted 10012, 10030, and 10059 times, respectively, which is equivalent to one-hundredth homeopathic dilution

C12, C30 and C50, or diluted 100727, 10030 and 100200 times, respectively, which is equivalent to hundreds of homeopathic dilutions of C12, C30 and C200. To prepare a solid dosage form, the solid carrier is treated with the necessary dilution obtained by the homeopathic process. To obtain a standard dosage form of the combination of the invention, the carrier mass is enriched with each of the dilutions. Both saturation orders are suitable for the preparation of the desired combination dosage form.

In a preferred embodiment, the starting material for the preparation of the activated potentiated form containing the combination of the invention is a polyclonal antibody obtained from an animal to the corresponding antigen, namely, interferon gamma or protein 5-100.

To obtain an activated potentiated form of polyclonal antibodies to gamma-interferon, the corresponding antigen can be administered as an immunogen to laboratory animals, if possible, to rabbits. Polyclonal antibodies to interferon-gamma can be obtained using whole interferon-gamma from the following sequence:

The sequence of ZE. I. MO;

Meg Was Tu Tv obeg o Tu Ne be Av Rie s Se was Ne Ma

From 5 to 10

Yvo su bag oivzy Su Suv Tug o Suv obi Avro bo Tu Mar oguv 16 250 sho 25 y 30

Aza st Ava ieo yuv Tu Tu Bbe Ava Abo su Niv bBeg Azv. Wow

From 35 to 45

Aa Avr Avp UM TP axis baye fev o byu Me be) Guz Aza Tr o Buv 45 50 56 ba

Sish would be Azr At Guz Ne Me Oip be obl Ne Ma! beg RNe 61 85 to 75

Tug ble Guk fesh Riv Guv Ap o RAes was 5 o Ava For ego He did not sit down 76 Vo 85 cha

Km Web Ma bin TpRONe iuv Siv Abro Mer Avo Ma uv Re Re 91 95 KU 3) 105

Avp o begoAva uv uz uv Ag der AvroRbe b yuv os Togo Av 108 o 5 120

Tug o Zeg UVI iVbg o drogetssh Aby Ua! b Robs Aa No Neo b 151 125 130 135

Bec Pe o bip Maii Mef Av bsh iyu Zeg bto Aa Aga iuv To bu 136 To: 145 150

Suk Ag Buv Ag beg siy Maebivch Re Ag bu AgoAg Aa Veg 151 155 180 155 sp 186

Polyclonal antibodies to interferon-gamma can be obtained using the following whole sequence of interferon-gamma:

EC sequence. Z. MO: 2

Mei Muvo tm Ti ego Tub Ne ate Av Re bib be Su Neo Ua! 1 5 KO 15 ey Siu beg ate Obmzh was Tub was byu Azr oto bus o Uai Was s te 20. 25 30

Aa si dva Mep iuv Sub Tu Rve Avp Av o OM Niv ego ABroma! 31 35 40 45

Aza Avr Aza OM ChbgoGeno Re be) su Ne Sei Guz Ba Ter Guk 46 5 55 50

Osh bSish Zeg o Avr Agu Sub Ne Meb Oto bBeg Op Ne Mag o Zeg Re 81 65 t 73

Tut Re uz iien Rne Tuz DAZp Re (uz Avr o Avr obi ber oNe zhe 76 0 85 0 iu5 Zv Ua ish TVg Ne fu bib Abro Mer AvpoMa! Tuz Re Re a 5 150 105

Avl Zeg o Avp Buv uv Buv Ak oAvroAvro Re Oh fuye bey Tvgbo vy 106 119 5 120

Tug Zeg Woah! TygoAurotsei Avp Mag obchni Agu was o Av o Ne Nivo osa 121 125 130 135 ii vo Ne b ma! Mei Av bab ii6m o Zeg Ro Aa Av uv ivgo vu 136 140 145. 159 uv Ag iiuz Aga be biv Mefoien Re bi bu Ag Ag Av o beye 151 155 180 165 vii 166

E It is also envisaged to use parts of gamma-interferon as an antigen: The corresponding sequence for such an antigen is instructive:

BO sequence. Yu. MO: Z

Did not eat Av: Rpe bib Ge o Suye Ne Ma 7 10 15

Kei EM Berg sen bm buye TU bu bip Azr o Rgo Tug May Zu8 bij to 25 Ko)

Aa OM you Te) Was in Tug RBE Avp o Az su Nib beg Avro Ma!

With | 5 40 ac

DE AvroAvp Ou Lit iv Reia was Ne 8 5O 55

ZEO sequence. 1Yu. MO: 4

Xia oAbrobge tugo Ma Buv dA Ko

Av bi Azp be fuz Tuz Tut Re Avya div bu Niv beg Avr Ma

And 35 at 45

Dido Avrodep o siu y fey Re vu Su Ne Gay fuz Av ytro was io B 55 bo bin biv beg Avr Aga Suv Ne Meb bij Zeg bij Ne Mai Bego Ra. in 5 KI 75

Tub Rnpe uv fe Re gub Ava Re gub AvroAvro bij ego Ne Z 75 va 85 0 uz Zeg mai bin Lygo Ne uv in Dvro Me Avpo Mag obu ble rpe chi 95 to 195

Avp o beg Avpofuz uv iuz Ag AvroAero Re Sino fuv Sei Tab Ava 16 o 5 129.

Tug Beg Mar Tig o ler oTiey Abp Ua! Oi Ag Su Av Na niv ots 21 125 130 135

Men Don't fight mag Me Aha and Seh ZCheg to Ai should fuz Tr Ou 136 140 145 150

Hugo yga Su Ag beg Oi Mei yiv) Be Ar obu Arr OA Av o Beg ii 58 tvo 155 ip 166

Sequence of BE. NU MOH Z bSip Ar Ro eat Mar osuye byu

Av o Si Aza Ge Tuz Tu tuye bBe 24 Av bu Mi beg Avr Zp dep y "chai

From AvroAvpo ZU 35 Rec Reven Siu 40 Tez iuv iza Ter 45

Aa Te Ne Guye bi Bek Abr B) 1uv Ne Mes o bi 55 bij Ne Ma Zeebye 50 o Av Veg Riv 81 Re uh Gay 85 (uv Azy Bra Gu 70 Avr Si beg Ne 75 tJ Re AB ek! 75 Zeg mag Zi 80 Ne yiub Ov ABro8uU Avp Ma! Suv Rpye va

Ge tyag Me Re «I Zeg oAvpoGuv 9 Gu5 Ag AeroAvro 00 bio Buv fe Tok o 105

Ap iub Re Av

BV oBeg Wow! TE 119 yeey Aza Ua! biv 15 in Av Ne oNiz (120

TUg Avgr Ag sy 191 125 1360 135 yesh ne bi Uvi Me Az bib Tets o beg Bo Av Av uv Theo Ou t38 340 145 lei yiuv Aga Guz Aeyudr obeg cha Meb iiey Re ip Su At Aga Aa bBeg 181 155 160 I 165 siy 186

Sequence of EARTH. MO: 5

Sp obBego bi Ne Mo bego Rae and 69 I and

Tu o Rije uv fey Re fuv Avpo Re Gu sAvroAvbro bi ego Ne bij 75 80 Ve vo

Tuz Zeg may Zv TL Ne fu bib Azr o Me Aza Ua was ble re

Thu 95 at 105

Avp o bek Abp Tu Buv Hugo AR) Avro Or Re so fu Ten TAE leg 06 10 in t

Tug Beg Ma 121 123

EC sequence. 10. MO: U

Me Azlo Waiiuye RBe Riv 100 5

Azy o BegABY ofub Guz Tuz Ag dvr Ar Riv si 1uzivts TE Av 106 o 5 120

TU begMa! Tr odAvr Geo Avpo Mar biy Agroiuz Abe Ni sshsh 12 125 130. ieu Nebi Uvi May Av o schi Mez Zeg Ryu 135

B 140 145

Sequence 5620. Yu. MO 8 century b Tig LeiyueOyu Ar Me Asp Ua uz Rve Ra 92 Fe 100 105

Ap oZegsAvp iuz uh fu Agro Avr Ar. Rae byu Guya Tseng Tr. Av tob for 5 120

ThugbagMma! TpgAvr Hey Av Ma! beep yo 21 125 30 g Sequence ZEO DU. MO 9 che Te o Avro Teo Ava Mar obed du uv Av o Ne Ni s 123 125 130 135 iv Ne st ouar Mer Ave oom iienobego Re Av o Aa 1368 149 145 147

Sequence of ZEE; MO: THAT

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KO sequence. 10. MO 11

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Avp o oBegoAva Buk o Guz Buv Ag Ar: Avr 106 119 114

Polyclonal antibodies to gamma-interferon can be obtained using a molecule of recombinant gamma-interferon from one of the following sequences;

EC sequence. U. MO 12

Mei biz Avr Re Tug Ma uv bi 28. Zo

Av Si Aza iey uv o iuv Tu Rpe dep Az bu H5 o Beg o dev Ma

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BI whatever 105

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Zp t66

The sequence of ZO, OO. MO: 13

Mei Si: Avro bio Tut Mai o iuya C 24 za

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A typical procedure for preparing initial polyclonal antibodies to human interferon gamma can be described as follows: 7-9 days before blood sampling, rabbits are given 1-3 intravenous injections to increase the level of polyclonal antibodies in the bloodstream. After immunization, blood samples are taken to detect the level of antibodies. As a rule, the maximum level of immune response to soluble antigen is reached 40-60 days after the first injection. After the completion of the first cycle of immunization, rabbits undergo a 30-day rehabilitation period, after which re-immunization is carried out with the help of 1-3 intravenous injections.

To obtain antiserum containing the necessary antibodies, the blood of immunized rabbits is taken and placed in a 50-ml centrifuge tube. Clots of the product formed on the walls of the test tube are removed with a wooden spatula, and a stick is placed to the clot in the center of the test tube. After that, the blood is placed in the refrigerator for 1 night at a temperature of approximately 40 °C.

The next day, the clot on the spatula is removed and the residual liquid is centrifuged for 10 minutes at 13,000 rpm. The supernatant is the necessary antiserum. The resulting antiserum is usually yellow in color.

2095 MamM»z (weight concentration) is added to the antiserum to obtain a final concentration of 0.0295 and before use it is stored in a frozen state at a temperature of - "C (or without the addition of McMeese - at a temperature of -70 "C). The following sequence of absorption on the solid phase is suitable for separating the required antibodies 20 to gamma-interferon from the antiserum: 10 ml of rabbit antiserum is subjected to a two-fold dilution of 0.15 M mass, after which 6.26 g of Mag5O" is added, mixed and cultured for approximately 12-16 hours at a temperature 4" C. The precipitate is removed by centrifugation, dissolved in 10 ml of phosphate buffer and dialyzed against the same buffer for 1 night at room temperature. After removal of the precipitate by centrifugation, the solution is placed on a column of

DEAE-cellulose balanced with phosphate buffer. The antibody fraction is determined by measuring the optical density of the eluate at 280 nm.

Isolated crude antibodies are purified using the method of affinity chromatography by attaching the obtained antibodies to endothelial MO-synthase, located on

From the insoluble matrix of the medium for chromatography, further elution with concentrated water-salt solutions.

The resulting buffer solution is used as the starting solution for the homeopathic dilution process used to prepare the activated potentiated form of antibodies. The preferred concentration of the original matrix solution of antigen-purified rabbit polyclonal antibodies to gamma-interferon is 0.5 - 5.0 mg/ml, if possible, 2.0 - 3.0 mg/ml.

Brain-specific protein 5-100, expressed by neurons and glial cells (astrocytes and oligodendrogliocytes), directly or through interaction with other proteins, performs a number of functions in the central nervous system aimed at supporting normal brain functioning, including influencing learning and memory processes, growth and viability of neurons, regulation of metabolic processes in neuronal tissues, etc. To prepare an activated potentiated form of antibodies, antiserum to brain-specific protein 5-100 can be removed from brain tissue and processed as follows: - bull brain tissue frozen in liquid nitrogen is turned into powder using a special grinder; - proteins are extracted in a ratio of 1:3 (mass/volume) using an extracting buffer with homogenization; - the homogenate is heated for 10 minutes. at a temperature of 60 "C, then cooled to 4 "C in an ice bath; - heat-labile proteins are removed by centrifugation; - fractionation with ammonium sulfate is carried out in stages, with subsequent removal of precipitated proteins; - the fraction containing protein 5-100 is precipitated with 10095 saturated ammonium sulfate, which is carried out by lowering the pH level to 4.0; the necessary fraction is collected by centrifugation; - the precipitate is dissolved in a minimum volume of buffer containing EDTA and mercaptoethanol, the precipitate is dialyzed with deionized water and lyophilized; - after fractionation of acidic proteins, chromatography is carried out on an ion-exchange medium - DEAE-cellulose OE-52, and then DEAE-sephadex A-50; - collected and dialyzed fractions containing 5-100 proteins are separated according to

From the molecular weight by gel filtration on Sephadex 5-100; - purified protein 5-100 is subjected to dialysis and lyophilized.

The molecular weight of the purified brain-specific protein is 5-100 - 21000 0.

Polyclonal antibodies to protein 5-100 can also be obtained using a technique similar to that described for antibodies to gamma interferon, using an adjuvant. A whole molecule of protein 5-100 can be used as an immunogen (antigen) for immunization of rabbits.

Buchanya ZZOTSV ECHO 10. MOK 14) whole soybeans As food We eat wild wild beans; Mich

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Lkdesy BYKA (KP. NM MOSTU

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Gkodivka ZHTOOg PZEOM MO OMO 16) era Shcheg si iv sm ok oma ma be Te iano no lap hv ke ne AD ne stou My su derbi Ly obue yem ve k 75 SCHE KO tsu uh bi izheideu zhi ev s peiya She obhu rizh 31 KN cho ah ie Akhrofmh bit o zue aero Me! Ah) Me o UarAero tsu MM UZH uv ke sh s Ba tk Bso kros yo su ABrOMu S che vu oi shehe He ta mo sheizy Mo Me izui TVO MU Ou lez deya RUEOORNE te 8 I ate

Chtrobyam av Be a - fizimo BA LIBVEOM MOMOT

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Ki Z lo Ah inmoman oko (yo ii MHerolyuYu MM: lii o mmearoh zuye liu MO ih "o schi t y sSfino Aurobyu dovi dorsu Me m'sryyuu rne s YAM tuye t tk 73 ta km: iv MM o vy DU di TM heh o Mn luUM Ah I give them ryu two more bah k 5 cho

To obtain antiserum, brain-specific 5-100 protein or a mixture of protein 5-100 (antigens) is prepared in combination with ethylated bovine serum albumin as a carrier substance with complete Freund's adjuvant, and pre-isolated brain-specific protein 5-100 is added, which is injected subcutaneously into the experimental animal. (rabbit) in the back area in a volume of 1-2 ml. Antiserum can have a titer of 1:500-1:1000.

The activated potentiated form of each component of the combination can be prepared from the original solution by homeopathic potentiation, if possible - by the method of proportional reduction of concentration by a series of dilutions of 1 part of each previous solution (starting from the original solution) in 9 parts (for decimal dilutions) or in 99 parts (for hundreds of dilutions), or in 999 parts (for dilutions in a thousand - attenuation M) of a neutral solvent, starting from the concentration of the original antibody solution in the solvent, if possible, water or a mixture of water and ethyl alcohol, in the range from approximately 0.5 to 5.0 mg/ml, in combination with external influence. If possible, the external influence should include repeated vertical shaking (dynamization) of each dilution. If possible, separate containers should be used for each subsequent dilution to the required activity level or dilution factor. This method is generally accepted in homeopathic practice

See, for example, V. Shvabe "Homeopathic Medicines", M., 1967, p. 14-29, which is incorporated herein by reference for its stated purpose.

For example, to prepare 12 percent dilutions (denoted as C12), one part of the original matrix solution of antibodies to gamma-interferon with a concentration of 3.0 mg/ml is diluted in 99 parts of a neutral water or water-alcohol solvent (if possible, 15 95 ethyl alcohol). , and then vertically shaken many times (10 or more) to prepare the 1 hundredth dilution (denoted as C1). The 2nd hundredth dilution (C2) is prepared from the 1st hundredth dilution C1. This procedure is repeated 11 times to prepare the 12th hundredth dilution of C12. Thus, the 12th hundredth dilution of C12 is a solution obtained by 12 successive dilutions of one part

From the original matrix solution of antibodies to brain-specific protein 5-100 with a concentration of 3.0 mg/ml in 99 parts of a neutral solvent in different containers, which is equivalent to a hundredth homeopathic dilution of C12. Analogous procedures with the appropriate dilution factor are carried out to obtain C30, C50 and C200 dilutions. Intermediate dilutions to test activity can be tested in the required biological model. The preferred activated-potentiated forms for both antibodies of the invention are a mixture of C12, C30 and C200 dilutions or C12, C30 and C50 dilutions. When using a mixture of different homeopathic dilutions (first of all, hundreds) of the active substance as a biologically active component, each component of the drug (for example, C12, C30, C50, C200) is prepared separately according to the above procedure, until the penultimate dilution is obtained (for example , to C11, C29, and C199, respectively), and then one part of each component is added to one container based on the mixture of the drug and mixed with the required amount of solvent (for example, with 97 parts for hundredth dilutions).

It is possible to use the active substance in the form of a mixture of other different solutions according to homeopathic technology, for example, decimal and/or hundredths (020, C30, C100 or C12,

C30, C50 or C12, C30, C200, etc.). Their effectiveness is determined experimentally, by testing the dilution on a suitable biological model, for example, on the models described in the examples below.

External treatment in the course of potentiation and reduction of concentration can also be carried out by means of ultrasonic, electromagnetic or any other physical influence accepted in homeopathic practice.

If possible, the combined medicinal product of the present invention may be in liquid form or in a solid standard dosage form. A preferred liquid formulation is a mixture, preferably 1:71, of an activated potentiated form of anti-gamma interferon antibodies and an activated potentiated form of anti-protein 95-100 antibodies. A preferred liquid carrier is water or a mixture of water and ethyl alcohol.

The solid standard dosage form of the drug according to the present invention can be prepared by impregnating a solid carrier, acceptable in pharmaceutical practice, with a mixture of activated potentiated form of aqueous or aqueous-alcohol solutions of the active components, mixed, if possible, in a 1:1 ratio. Alternatively, the medium can be enriched sequentially with each appropriate dilution. Both saturation orders are acceptable.

If possible, the medicinal product in a solid standard dosage form is prepared from granules of a carrier acceptable in pharmaceutical practice, which has been previously saturated with water or water-alcohol dilutions of the activated potentiated form of antibodies to gamma-interferon and the activated potentiated form of antibodies to protein 5-100. Solid medicinal

The form may be in any form known in pharmaceutical practice, including tablets, capsules, lozenges, and the like. Glucose, sucrose, maltose, starch, isomaltose, isomalt and other mono-, oligo- and polysaccharides used in the production of medicines can be used as inactive pharmaceutical ingredients, as well as technological mixtures of the above-mentioned inactive pharmaceutical ingredients with other auxiliary substances acceptable in pharmaceutical practice, for example, isomalt, crospovidone, sodium cyclamate, sodium saccharin, anhydrous citric acid, etc.), including glidants, disintegrants, binders and dyes. Preferred carriers are lactose and isomalt. The pharmaceutical dosage form may additionally include standard pharmaceutical excipients, for example, microcrystalline cellulose, magnesium stearate and citric acid.

To prepare a solid form for oral use, 100-300-μm lactose granules are enriched with aqueous or water-alcohol solutions of activated potentiated form of antibodies to histamine, activated potentiated form of antibodies to gamma-interferon, and activated potentiated form of antibodies to protein 5-100 in the ratio of 1 kg of a solution of antibodies per 5 or 10 kg of lactose (1:5-1:10). In order to saturate the lactose granules, they are subjected to saturation irrigation on a fluidized bed at a suitable installation (for example, "NashIip

Riyutsar" manufactured by Nynip ZtrN) with subsequent drying with a flow of heated air at a temperature below 40"C. The prescribed amount of dried granules (10-34 parts by mass) saturated with the activated-potentiated form of antibodies is placed in a mixer and mixed with 25-45 parts by mass of "unsaturated" pure lactose (which is used in order to reduce costs, simplify and accelerate the technological process without reducing the therapeutic efficiency), together with 0.1-1 mass parts of magnesium stearate and 3-10 mass parts of microcrystalline cellulose. The resulting tablet mass is homogeneously mixed and tableted by the method of direct dry pressing (for example, on a Kogesp-XI. 400 tablet press) to form 150-500-mg round tablets, if possible, 300-mg. After tableting, tablets of 300 mg are obtained, which are saturated with a water-alcohol solution (3.0-6.0 mg/table) of a combination of activated potentiated form of antibodies to gamma-interferon and activated potentiated form of antibodies to protein 5-100. Each component of the infusion combination is in the form of a mixture of 100% homeopathic dilutions of C12, 60% C30, and C50 or a mixture of 100% homeopathic dilutions of C12, C30, and C200.

While the invention is not limited, it is believed that the activated potentiated form described herein does not contain the molecular form of the antibody in an amount sufficient to provide the biological activity attributed to such molecular form. The biological activity of the combined agent (combined drug) according to this invention is sufficiently fully demonstrated in the attached examples.

In addition, the present invention represents a method of significantly delaying the onset of symptoms of multiple sclerosis, namely, a method that includes the administration of a combined drug containing a) an activated potentiated form of an antibody to gamma interferon and B) an activated potentiated form of an antibody to protein 5-100 .

In addition, the present invention represents a method of reducing the probability of relapse in patients suffering from multiple sclerosis by administering a combined medicinal product containing a) an activated potentiated form of an antibody to gamma interferon and B) an activated potentiated form of an antibody to protein 5-100 .

If possible, for the purpose of treatment, the combination according to the present invention is administered from 1 to 4 times a day, if possible twice a day, 1 or two combined standard dosage forms at each reception.

The invention is further illustrated by the following non-limiting examples.

EXAMPLES

Example 1

Female Wistar rats (200-220 g) were used in this study. EAE was induced in them by a one-time cultivation of an encephalitogenic mixture based on 100 mX homogenate of the homologous spinal cord, 0.2 ml of CEA (content of destroyed mycobacteria - 5 mg/ml) and 0.2 ml of physiological development in one animal. E55 was injected subcutaneously (at the base of the tail along the tail vein) under anesthesia with ether in the amount of 0.4 ml (0.2 ml on the right and left) (Abdurasulova, IN et al., 2004; Zhitukhin Y.L. et al., 2008; Serebryana, N. .B. et al., 2010). The following drugs were administered intragastrically 2 times a day at 7-hour intervals for 30 days, starting from the day of activation of EAE: (a) Ultra-low dose of antibodies to protein 5-100 (further on in the text - "NND abs. to protein 5-100") (p-10, 2.5 ml/kg/d); (5) ultra-low dose of antibodies to gamma-interferon (further in the text - NLD abs. to interferon-gamma") (p-10, 2.5 ml/kg/d); (c) combination of NLD abs. to protein 5-100 and NND abs. to interferon-gamma (hereinafter referred to as "combined drug") (n-10, 5 ml/kg/d) and (a)

From distilled water (control; p-10, 5ml/kg/d). The immunomodulator Copaxonof) (Tema, Israel) was used as a reference drug, which was administered intramuscularly at a dosage of 4 mg/kg, 3 2 to 25 days after EAE activation. The results are shown in fig. 2-5.

Clinical symptoms of experimental autoimmune encephalomyelitis (EAE) were assessed every day for 30 days, starting from the day of activation of experimental autoimmune encephalomyelitis (EAE). The examination of each rat was carried out immediately before the introduction of the studied drugs. In the case of rapid disease progression, clinical symptoms were evaluated twice a day.

Severity of neurological abnormalities was assessed in points: presence of muscle weakness, tremor (0.5 points); resistant paresis (1 point); paralysis (1.5 points).

The clinical index (CI) was calculated as the sum of symptoms in 4 limbs. In addition, SI was scored as zero if there were no visible clinical signs of neurological abnormalities, and scored as 6 in case of animal death. The maximum SI value was recorded every day. The cumulative index for each rat, which was calculated as the sum of individual CIs during the entire disease period (30 days), was also recorded. The 30-day period of the disease was divided into the following phases: latent phase (1-10 days); phase of clinical manifestations (11-18 days); and the recovery phase (19-30 days).

In order to evaluate the effectiveness of the studied drugs in the model of experimental autoimmune encephalomyelitis (EAE), the following parameters were analyzed in each studied group: 1) time of disease onset (days); 2) changes in the average severity of the disease over time (SI, scores); 3) the average severity of the disease at different phases (SI, points) 4) the total number of relapses, return of symptoms of the disease (95).

NND abs. to protein 5-100 delayed the onset of clinical symptoms of disease periods, both in comparison with the negative control (p"0.05) and in comparison with Copaxone (p"0.05). The onset of the disease in the two groups receiving NND abs. to interferon-gamma, and in the group receiving Copaxone, it did not differ from the control (see Fig. 2). At the same time, in animals administered the combined drug, there was a tendency to shorten the period of clinical signs of multiple sclerosis in comparison with the control. It should be noted that there were statistically significant differences in the time of onset of the disease between the group receiving NND abs. to protein 5-100, and the group receiving the combined drug. because the percentage of animals with a severe course of the disease (73 points) was also lower in the group receiving NND abs. to protein 5-100. In the groups receiving the combined drug,

the percentage of animals with a severe course of the disease was higher than in other groups in all periods of the disease (see Fig. 3).

Due to the fact that different mechanisms of the pathogenesis of EAE were activated in different phases of the development of multiple sclerosis, an analysis of the effect of drugs on the severity of the disease in the latent period, in the phase of development of clinical signs and in the recovery phase was carried out. Introduction of NND abs. to protein 5-100 significantly contributed to the reduction of the average scores of the manifestation of clinical signs of the disease in the phase of clinical manifestations and in the recovery phase, while Copaxone and NND abs. to interferon-gamma significantly reduced the severity of clinical symptoms only in the last phase. The combined drug, in comparison with the control and groups that received only NND abs. to interferon-gamma or only NND abs. to protein 5-100, significantly increased the average scores of exposure to animals in relation to the appearance of clinical signs at the stage of the disease and recovery (see Fig. 4).

It should be noted that in some animals, after the complete disappearance of clinical signs of the disease, a state somewhat similar to a relapse was observed, i.e., the return of clinical manifestations of EAE. In the group receiving NND abs. to protein 5-100, despite the favorable effect of the drug on the clinical course of EAE (later period of disease onset, lower severity of clinical signs), a larger number of animals (25,905) with relapses was noted. At the same time, in the group that received the combined drug, which was characterized by an early onset of the disease and more significant manifestations of the pathological process, there were no relapses in any animal. (see Fig. 5).

Conclusions:

In the model of experimental autoimmune encephalomyelitis (EAE) NND abs. to protein 5-100 had a beneficial effect on the course of the disease. NND abs. to protein 5-100 reduced the clinical signs of the disease at its onset, as well as the severity of the symptoms. NND abs. to interferon-gamma and a comparative drug -

Copaxone - did not affect the course of the disease in the selected group

From the experimental model of multiple sclerosis, that is, when using these substances, the amount of clinical signs of the disease periods and the severity of the symptoms did not differ from the control (distilled water).

The manifestation of the effect of the drugs depended on the stage of development of EAE (latent phase, phase of appearance of clinical signs, phase of recovery).

The effect of the combined drug differed from the effect of the components, namely, separately NND abs. to protein 5-100 or NND abs. to interferon-gamma. The use of the combined drug led to an increase in the immune response. The combined drug led to an earlier onset of the disease, an increase in the number of sick animals and an increase in the severity of the disease.

In 25 95 animals in the group receiving NND abs. to protein 5-100, and 15 95 animals in the group receiving NND abs. to interferon-gamma, relapses occurred during the study period (30 days). At the same time, relapses were not observed in any animal in the group receiving the combined drug. The most typical course of multiple sclerosis is the relapsing-remitting subtype, characterized by unpredictable attacks (relapses), followed by periods of relative remission without new signs of disease activity.

The primary goals of multiple sclerosis therapy are to reduce neuronal damage during acute attacks and restore function after attacks, as well as prevent new attacks that can lead to incapacitation.

NNDabs. to protein 5-100 alone were able to improve the clinical symptoms of EAE during all phases of the disease model, making them potentially useful for controlling multiple sclerosis attacks. The combination of NND abs. to protein 5-100 I NND abs. to interferon-gamma completely prevented the return of clinical symptoms of EAE and, thus, can be used in the therapy of multiple sclerosis in humans to prevent relapses.

Thus, the combination of NND abs. to protein 5-100 4 NND abs. to interferon-gamma is a promising drug for the treatment of multiple sclerosis, which is believed to promote more effective rehabilitation and prevention of relapses by enhancing the immune response at the peak of the disease. 60 Example 2

WITH

The sigma-1 receptor (01) is an intracellular receptor localized in cells of the central nervous system, cells of most peripheral tissues, and cells of the immune component. It is believed that this receptor, by controlling the homeostasis of intracellular calcium, regulates intracellular signals, leading to the activation of the relevant transcription factors and the transcription of the entire gene family. The sigma receptor is involved in the pathogenesis of various diseases, including infectious and neurodegenerative conditions. In this regard, drugs that affect this receptor and the effectiveness of the interaction of ligands with this receptor can be considered effective drugs for the treatment of neuroinfectious and neurodegenerative diseases.

The effect of the combined drug, which includes ultra-low doses of antibodies to protein 5-100 (a mixture of homeopathic dilutions C12-C30-C50) and ultra-low doses of antibodies to interferon-gamma (a mixture of homeopathic dilutions C12-C30-C50) in a ratio of 1:1, as well as components of the composition (ultra-low doses of antibodies to protein 5-100 (mixture of homeopathic dilutions C12-230-050) (NND abs. to protein 5-100) and ultra-low doses of antibodies to interferon-gamma (mixture of homeopathic dilutions C12-C30-C50 ) (NND abs. to interferon-gamma)), was analyzed by ip myo upon binding of the standard ligand NI|Ipentazocine with recombinant human 01 receptor and evaluated using the radioligand method. Potentiated distilled water (a mixture of homeopathic dilutions C124C30-C50) (potentiated water) was used as a control. 20 μl of the combined drug or 10 μl of NND abs. to protein 5-100 or NND abs. to interferon-gamma was added to the incubation medium. Thus, the amount

NND abs. to protein 5-100 and NND abs. to interferon-gamma transferred to the test well during the trial of the combined drug was identical to the amount of NND abs. to protein 5-100 and NND abs. to interferon-gamma, which were tested as monopreparations, which made it possible to compare the effectiveness of the combined preparation and its individual components that make up the complex preparation. Potentiated water was added to the incubation medium in quantity

From 20 μl and 10 μl.

Next, 160 μl (approximately 200 μg of protein) of a homogenate of the membranes of the Uigkai cell line (a line of human leukemic T-lymphocytes) was added, and finally 20 μl of a tritium-labeled radioligand ("NIpentazocine (15 nm). 32

In order to measure nonspecific binding, 20 μl of unlabeled ligand - haloperidol (10 μm) was transferred to the incubation medium instead of drugs or potentiated water.

Radioactivity was measured using a scintillometer (Torsoshpi, RasKaga) and scintillation mixture (Misgoz5sipi 0, RasKaga) after cultivation for 120 minutes at a temperature of 22°C in 50 mM Tris-HCl buffer (pH-7.4) and filtration using glass fiber filters (CE (B, RasKkaga) Specific binding (in test or control) was calculated as the difference between total (in test or control) and non-specific binding.

The results (when measuring covalent binding) are presented as a percentage of specific binding in the control (distilled water was used as a control) (see Table 4).

Table 4

The influence of the studied drugs and potentiated water on the binding of the standard radioligand NIpentazocine to the recombinant human sigma-1 receptor and Quantity, о и , p. and binding

The studied group of specific binding of radioligands 2: introduced into the well. of radioligands in the control: the control of the bath 1 Z le (ee r indicator of the preparation of the wound of the tube)

Fo of specific binding in the control - (specific binding during the test / specific binding in the control)" 100,905;

Fo inhibition of specific binding in the control - 100 95 - (specific binding during the test/specific binding in the control) " 100 95).

Inhibition greater than 50 95 indicates the existence of a significant effect on binding.

A suppression of 25 95 to 50 95 confirms a minor to moderate impact. Inhibition of less than 25,956 is considered insignificant relative to the effect on binding and is within the original threshold values.

Conclusion:

C3

The combined drug more effectively inhibits the binding of the standard radioligand

GNIpentazocine with recombinant human sigma receptor than its individual components (NLD abs. to protein 5-100 or NND abs. to interferon-gamma).

NNDabs. to the protein 5-100, transferred to the test well in a volume of 10 μl, inhibit the binding of the standard ligand ("NIpentazocine" to the recombinant human sigma-1 receptor, but the intensity of this effect is less than when using the combined drug.

NND abs. to interferon-gamma, transferred to the test well in a volume of 10 μl, did not affect the binding of the standard ligand ("NIpentazocine" to the recombinant human sigma-1 receptor.

Potentiated water, transferred to the test well in a volume of 10 μl or 20 μl, did not affect the binding of the standard ligand ("NIpentazocine" to the recombinant human sigma-1 receptor.

Claims (8)

FORMULA OF THE INVENTION 1. Combined medicinal product for use in the therapy of neurodegenerative ZO diseases, containing a mixture of solutions of homeopathically potentiated forms of a) antibodies to gamma interferon in a mixture of three homeopathic dilutions C12, C30 and C 200 or C12, C30 and C50, and B) antibodies to protein 5-100 in a mixture of three homeopathic dilutions C12, C30 and C 200 or C12, C30 and C50. 2. Combined medicinal product according to formula 1, which differs in that the antibodies in the activated potentiated form to gamma-interferon are antibodies to the whole gamma-interferon, which has the sequence zZEO IU MO: 1 or for 5EO IU MO: 2. 3. The combined medicinal product according to claim 1, which is characterized by the fact that the antibodies in the activated potentiated form to gamma-interferon are antibodies to a part of gamma-interferon having sequences selected from the group consisting of ZEO IU MO: 3, ZEO IU MO: 4, BEO IU MO: 5, 5EO IO MO: 6, 5EO I MO: 7, 5EO IU MO: 8, 5BEO IO MO: 9, 5EO I MO: 10, 5EO IO MO: 11, 5EO IO MO: 12 and 5EO IO MO: 13. 4. The combined medicinal product according to claim 1, which is characterized by the fact that the antibodies in the activated potentiated form to protein 5-100 are antibodies to whole bovine protein 5-100. 5. The combined medicinal product according to claim 1, which is characterized by the fact that the antibodies in the activated potentiated form to the protein 5-100 are antibodies to the entire protein, which has the sequence 5-100 with ZEO IU MO: 14. 6. Combined medicinal product according to formula 1, which differs in that the antibodies in the activated potentiated form to the protein 5-100 are antibodies to the whole protein, BO, which has the sequence 5-100 with ZEO IU MO: 17. 7. Combined medicinal product according to claim 1, which is characterized by the fact that antibodies in an activated potentiated form to gamma interferon are presented in the form of a mixture of homeopathic dilutions C12, C30 and C50, impregnated on a solid carrier, and antibodies in an activated potentiated form to a protein 5-100 are presented in the form of a mixture of homeopathic dilutions C12, C30 and C50, subsequently impregnated on a solid carrier. 8. Combined medicinal product according to claim 1, where antibodies in an activated potentiated form to gamma interferon are presented in the form of a mixture of homeopathic dilutions C12, C30 and C200, impregnated on a solid carrier, and antibodies in an activated potentiated form to protein 5-100 are presented in the form of a mixture of homeopathic dilutions C12, C30 and C200, subsequently impregnated on a solid carrier. 9. Combined medicinal product according to claim 1, which is characterized by the fact that antibodies in an activated potentiated form to protein 5-100 are presented in the form of a mixture of homeopathic dilutions C12, C30 and C50, impregnated on a solid carrier, and antibodies in an activated potentiated form to gamma-interferon are presented in the form of a mixture of homeopathic dilutions C12, C30 and C50, subsequently impregnated on a solid carrier. 10. Combined medicinal product according to claim 1, which is characterized by the fact that antibodies in an activated potentiated form to protein 5-100 are presented in the form of a mixture of homeopathic dilutions C12, C30 and C200, impregnated on a solid carrier, and antibodies in an activated potentiated form to gamma-interferon are presented in the form of a mixture of homeopathic dilutions C12, C30 and C200, subsequently impregnated on a solid carrier. 11. Combined medicinal product according to claim 1, which is characterized by the fact that the antibodies in the activated potentiated form to gamma-interferon are monoclonal, polyclonal or natural antibodies. 12. Combined medicinal product according to claim 11, which is characterized by the fact that the antibodies in the activated potentiated form to gamma-interferon are polyclonal antibodies. 13. The combined medicinal product according to claim 1, which is characterized by the fact that antibodies in an activated potentiated form to gamma-interferon are prepared by successive hundredfold dilutions in combination with shaking of each dilution. 14. The combined medicinal product according to claim 1, which is characterized by the fact that the antibodies in the activated potentiated form to protein 5-100 are monoclonal, polyclonal or natural antibodies. Zo 15. The combined medicinal product according to claim 14, which is characterized by the fact that the antibodies in the activated potentiated form to protein 5-100 are polyclonal antibodies. 16. The combined medicinal product according to claim 1, which is characterized by the fact that antibodies in an activated potentiated form to protein 5-100 are prepared by successive hundredfold dilutions in combination with shaking of each dilution. 17. The method of treating a neurodegenerative disease, and the specified method provides that the corresponding patient is simultaneously administered a mixture of solutions of homeopathically potentiated forms of a) antibodies to gamma interferon in a mixture of three homeopathic dilutions of C12, C30 and C200 or C12, C30 and C50, and B) antibodies to protein 5-100 in a mixture of three homeopathic dilutions C12, C30 and C 200 or C12, C30 and C50. 18. The method according to claim 17, characterized in that said neurodegenerative disease is multiple sclerosis. 19. The method according to clauses of the formula 17 or 18, which is characterized by the fact that the indicated antibodies in the activated potentiated form to gamma interferon and the indicated antibodies in the activated potentiated form to protein 5-100 are administered in the form of a combined medicinal product. 20. The method of reducing the frequency of relapses in patients suffering from multiple sclerosis, which is characterized by the fact that the combined medicinal product according to formula 1 is administered. 21. The method according to formula 19, which differs in that the combined drug is administered in 1-2 standard dosage forms, and each dosage form is administered from 1 to 4 times a day. 22. The method according to formula 19, which differs in that the combined drug is administered in 1-2 standard dosage forms, and each dosage form is administered twice a day. 23. Medicinal preparation for use in the therapy of patients suffering from multiple sclerosis, and the indicated preparation is obtained by combining solutions of homeopathically potentiated forms of a) antibodies to gamma interferon in a mixture of three homeopathic dilutions C12, C30 and C200 or C12, C30 and C50 , and B) antibodies to protein 5-100 in a mixture of three homeopathic dilutions of C12, C30 and C200 or C12, C30 and C50, each of 60 of which is prepared by successive multiple dilutions and multiple shaking of each resulting solution according to homeopathic technology and then or combining potentiated solutions by mixing them, or, alternatively, by impregnating the carrier with the specified combined solution or solutions separately. 24. The method of treating a neuroinfection associated with damage or destruction of nerve cells, which is characterized by the fact that the specified method involves the simultaneous administration to the corresponding patient of solutions of homeopathically potentiated forms of a) antibodies to gamma interferon in a mixture of three homeopathic dilutions C12, C30 and C200 or C12 , C30 and C50, and p) antibodies to protein 5-100 in a mixture of three homeopathic dilutions of C12, C30 and C200 or C12, C30 and C50. 25. The method according to claim 24, which is characterized by the fact that said antibodies in activated potentiated form to gamma-interferon and said antibodies in activated potentiated form to protein 5-100 are administered in the form of a combined drug. how else ; , - a list of SEQUENCES ot zlyu sad tat tii kesh ov ku her B I om she ee "tSHOOoz EPShtTEIV Oleg o ILYECH sr seii, bozhets BIBIEV shi em mom r IT TATI KITIV zidenttyi rt VE "zo" COMBINED BAVMACEVTIC COMOCHINIA AND METHODS OF BAKING slot z . rya EC account TT ST sieve JEEBNLO COMBINATION soot TVO MKU and tu chkoh pe uzhr vni M rus sv xx TE "be toi bure"" protein; for Fran dulyu koi ev torkanism-A nom Weight etb "ab 1 Mmek GBud tTUue Tvy shah Tuk dia Bbem Mi be Siv Bez Suv YIT1ie Uk" Gay 1 | Be 18 5 U "g s from Kasa uko dna RIDI schu puh tik Di Zola DIKE pk ZHI shiUu at yin SM Kuk Tuk was kpolLeyuU already tuye Maya was SYA AJ Si Y Y rey Y ih o y 30 myh pore teku p p do dya potiiy Da sg pod TE action vad liu wat yny Be iUh Huk vne dat Asia y u nyka UVO cha1i dka Er AV o zo 25 ich t vyny s and I t that I Z lovu Iz i sk eh bit tlaE this obreee This biu ie lu was beating tTerobu svis sl be dey For BA ay I and ee, . 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ЖАВсз. water kre so dredyu Mokh iz "x Shut Buv liy laer Mei llu" Uai Gu EMe Bpye azy bat oynp was RGme was tu ni to y dih "sia that 145 TO. ч чк вк г Ya zr" h ko Pukh vk Pav sAktu o Tochal Doju. ha vzrvoyvo rye sich bu iye: TVzh yza o Tukhk Shchek Mai Turgo deroCses dat uv 115 255 125 Iii KA uhe ;: z Sho taya shchi tru a kad che Eat together Bush Aza o Tiv NIiB Siz Bey Ti biy Udi Mes Ata Su fei; Bek i as vd t i35 140 pu tr pt, Z N ki m zhi - Tok uchy - TK Anyu sokah in, Dok Ego oATa via Buv Tak osiu was Ata Mu As ber yaziIy Me obo) vve dk chi45 150 155 t5o pi vh Akea doyin vek bi 1645 "1" is "what 15 OK be LADY "183" Yaoto Zariele "Eh ni o KU ptrKi stychy zhevu g chiiz SOURCE PSKOVZENATION deti tru: keshzhshi 1.55 psy pa nn yam ytei suyuen prezhvehn: pu with student z ra r serreniyame-" Nemo zariechtiv "Zh I g I NYA ggev chi. polu, that ie tod that en AN yar vich then Me Buv oOTuk ALSO bek bik Iyiv cezoAiz Epe bij obeb Suv shde UschA femz : и и; 18 te and ShcheYa a ko bi pet this siu was Tuh Suz bipoivr ro Tuk Ma) Bu biz A1ia sich schu 25 Zo Dvpogvo Buvoiyuya tus Bpe Av o Aia b1iu Shchi bek dvo Uuaii Aza Avo Dap Z 40 43 biu Tv oce) Ve ber biu ii BCeb Buv Depo Ter buh pod bii bek dvo o 5 va Akch was Yi1e Mei biv Chek biv T1v Uvi bek RBe here EBe Muvo Temp pe 7 75 vo Buv yzp Be Buk dr Azr bil beh YITte bip buh bZeh Udi bi tTVg her B5 y вх Муз sela Avr Me Aa Uki ime Ble Be Avgpobek ABY Buv Bu Buz DF i0o lov her deroa5v pe bim uv bem TBK Avi Tuk bej UyaV tri Avr oi av May shko: 12 125 si» AK Tuz Av ttie Niv ovi lei giv bij Uzi MV action 5105 Pem bek 80 135 Io Rgo s dz zha Bu tp btU buye yru pu Azh bek bii MeE and Rie SIY 145 159 155 i160 TU Aga Aa AT Sheh S 5 "210" Z "811 45 "212" VNT "ka" Chocho pariv kveI" zayi" JEWEL OF APPRECIATION "in" 1.43 taya tou buree"norotein" Isergavism-"Noto variv" "ap" tt yivz ATZ ve biv isp buye TV Mav tsev slya; eh em stu Su tut i 5 Hey 45 kUv'siv dayu ro Tuk chai Mu BIO piv io Av Yi Bu was Tuyu tse: ho | 28 Zo av Aa spiuU niv bek Avrouzi Aiv Ar oAvBo bu Tu Gay o vBestyan ZU Z yap 45 TIB "21054 "8112 743 raw" RVE "thie" Moto brewing "their "ishiv TZHEEELO VOKHODAENYA "Vuz ii. 13 "ori" la) purae"pretehv" isoganisme"Noto vavrivlE" "poh y biv Avrobso Tuye Ua) ruz biz Akta bib Dyam Theo was Buye Tut Ryv yapc dia bi Cizh Shek AvroUvi Asia yzr ydi bij TVkobaem Be tseo Pit 118. 7 5 30 ivo PuzoAzv Tkrotsuya Shchi" io beth Ayar yko uv (from MEZH was o beyu 3 cha a Iie bab rey ve tuye Re Buv Bez VI Buv dv be uz dr oAZYU SIV BE b Chak Ii bij Buz Bak in 5 tp o Biv buv si: Akr Met dv uva muz | 73 75 and Ra rde Avposhakh dai Gu ruk uv Ka AVOo AYU RNe si; I have been to Tuk bek vy t oABOO Bey Ani Ma sim deka ev o Ata tiv Nv asla 159 195 110 Sed i1e Siv Uvi Meh Asia bid be bei Rgo biz AtTa pu TB biu gru herye5 129 195 yka Music ygo back siv Meb vu Re Dgo biu Ago ygo Aza bek bil 135 135 149 "9105 5 -91st 143 "im vt zhav Nome zara "she kesh" SOURCE POHOPZHEYNYA "Hey her "ia o yvei Surezh"proteiy" lovrganisme"shchove variela" "4032 u piyoazhev REO Tut tai BbubB o bti action bij dB gay PUu8 Buv Tuk RpE de Z Z 15 15 Vi biUu Ni Be AvroUgi vAia ar aivy 0iUu TV ia No (o BIU TT A. Z za tsem Was two o tTgr was Ok bis bek two pu Was 116 Mes bip Bek siy Tie Mai Vas RnestTek RBe Guv pea Rae ru dev Bpe bud der Avr o biv v Zo BY Bak tiv spi; Ta o Zek uai piyu TV her Bu sb» ylr o Me Avpouvi Buv ih o 15 that pe Be vp obeh Avvobuv Buv Muz Ak Var dar Ra 1 Vuz her fah U: va 55 vyy Zhuk SesoUuai Yes Or opei Ay UdI StivsAZha BUB ATa Ti niv syh i10o 05 Id Gezhcho liv b Maiyi Media ssha zhoh eh VEO Aza Aza was TNK bi Beh viz tra 125 wk was wk Bezh bij Meb mem ri si bi Ato vaga dia bik Bij nos 135 140 "ijhe "dm A kargy. RE "vii" Nopse varieva "eh de SOURCE OF ORIGIN "y 1..55 kiy" pi bure uroyetien" zorganisuee "Noto variepe" kah zip Shek ip her Zai Bek BOye Tuye Be was in Bi was two would BE 1 that 10 15 yvr Ar osiv dvy tTie bip Puv betst Chai bi TB fi buk Bii beyu Mee o 5 zo AzpoUai tsu Rpe Re Azv oBesg Av tsu pu GU Akva VlroAvo Re sg a o IZ uv Bei yBkoABa bug bas Uvi what 55 kra "211: 46 "2i2z RENT "VISA" ote vzvrisetya ze chyki" EARNED COOLING "ry" u. "javaR Yani duree" pratetn" lorganisme "Nota zchpriedyv" mes Aa Ua uz this Rce two oats vBy Buv tsu Buv ArUu Derodyar re y Z IV ih Si Buv this Tps At tuyu Bach UZH Tk deroivnyu Azy MV siv Aa Sbuh y 2 Za ATta TT oninssiv betyu iz6e her Uvi Meh: Aibd biz ey ba orso for 4 AB "10" 8 "au" 3 "k2i18z RT "13" Cheomo zartzepe s" "yk1ih SOURCE. LIVING "priv 1.39 "y ns sheree" protein" lpyushtamnuzme "Nome shtor eta" "00 V Bek ak from TV 118 bu Biy) AhroMech ABY Ua) Pukh RI RN Avt dec i Z | I 15 Avp oMmuv eye buh Ak OR gave Be Sich was pev" Ts lav tuyu Bek chai I 25 Zo ti Ar o pez Ai Uvi bib vu 35 KI Z "2133 5 "vaz RE "13 Nomo zarivvv "aga -gshiz JHE ORIGIN " pasha, kiy" ischoi Suree" protown" lerganisme "Novek havietiv" "40059 Uvi Tys Ave tseo aap o udi biv Ak Mu din yi1yi6 Ni biz Men Iyi5 b) 1 9 15 15 Uvi Megssodia bip des Best ge AT Atd yi5 keiikh 1 « it KT EV Moto zvarueiaz chiya haiyi" GERENE ORIGIN karykh. "2233 /schoi bura-"proteh vy. tlorkanizshe"Noto variepe" sadah CE Shazh Tukh IT Ivo Aza re bilopez Suv Ti-zh Mv3 yivc biu pek ped u From 5. she was 15 o Ttuk Suv settled vvroBro Ter ovi Buv bi; ATZ bir deya Bea Was at 25 ZO Tek VnNe two Asia shu Niv bek Anor Uvi 35 35 "il 11 kaz ki "ev REGH "kei3z Notpto varivaza BC "Fo" SOURCE OF IGNITION, svohl 1,2 "253" lLabi Hurek"protain "l/orgvanizme" Noyuo karivla" "ai" 11 half TVt her There was a battle Der oMesodap Uai Mu Re re dyaloyes AvpoGPuv I i: 15 i5 pu buh ko Avro Arch "ei "ih zhAi "Fah vit kriZ" shtuchni peospidovnael "ap chiniv JEENO ORIGIN ia E.V "gas" / schoi guraee"protems" lpoiek" recombinant human gammasichterov" lovyukanisme"artificial sopdsovnosts" sao U. 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Meb chi der orko Tuhk Uli Bu ich A1a bij Av" o beb Buv Buya tuyu Bie 1 5 15 15 dvpoAiv Siu Nizh Bek dvr Uvi Asia Ar Av J Tig obere Mei bu yiBa Pvo Buv Ap Ter obuv bij ish Zak Avr As Buz tie Me biv Bas Z5 40 5 sik Ti Zai Bezh tva tuyu re pu ek BChe was DIA Be uv Dvre OR ve E o siy eh yi biv zuz Bej Uuai SIYA TV Ge Buv BIZ Azv Met AvpoMai JAK to pe o muy EPe RY Avo vezh dep buh Mu BU Ak AV ABb Rke obich was Gay Z Be TEO Avp otuh tu Uudi Tpe Ar Pec ABap Uvi BI Asya o BUuz AKA pue niv, in 105 119 siy ate the same battle in Media bIi0 ii-eo bek bo div svid ev TveE b) 315 129 125 Tu Ah was Ara Bek obip Me pe) Beijha Siu Du Aka div back 15 13 135 145 "10" 14 "2115 92 "vii VE -FI13" Vovsatya teh "she SHZHESENO INJURY podny schi ho? Shoi Suree "protasies" yorkanizm-bnov: Bbavatsev" -4005 4 Me det bij iliez bib Buv div Uzi Uai Aza her i1ie Ar oUai Rre Kiv i Z Io 15 pali Kuk bek BI" Abu siy stu Avo puv Ni Buv zeyp Bu Bu dec bly i eV Zo ev sbuv 01z vaie AvpoAka bib Hey bes har rve Her Siv syu ge 35 "o ah was ЗB bilo ЕТv uzi Uvi AbVrobme Uvi Me siv TB hey dvo bbh Ar 50 5 B To pz these ptejio Z I in Z sapany se I e ti YAN I - she. schu Ave 5IUu 5 Se yzo Re biv bi) rue Me vba BbyeE Ua Aga Me To TB 5 tiv taj tk At Su NiBOFIU Rv re Ot Nu bich ve 5 "10" 15 "2115 95 sho muh tu mai "im RU "|z Pe yani ia ia seli sha ХВ "hey" corner same in. Three of them, you kiki" WILDLY REFRESHMENT "pev" 10:35 this tue bure" troetekhny" ; ryn yavyti sg tur hu sarvganvm-"Poshcheo variene "Oh 15 yshm Fu IL Zhdu rt v; en chn y Ne s ti dk Ky er ih Mezh eh sulya Bei sii duv o Aiv Me Uvi Aia tsez Se Avro uv ke NEV i SE KZ ! SHO shi TUYE run TU run im slu AzhU UVO NIV Gu em was bue VEZ SO I | Me; Zo provuta se KU t op donya i UK enNae pokoy TYA u nya SHY them (Sk t wl ze og»uv 51 ipez oti Av dvy bi fer eh BivB RBe Bevobiy it Ite U 30 13 pUuz biz piv siv Ua Uzi Varomdeuv zi Mebi) TV pez Avrodvoa Ave ZO 55 v yti meha y uyu ky u ruta duvatch that 3 A EN tourom U ha yayu iUu AvbosiUu bil oSuv yzo Be bio b3v re mes AtTa svya U) ALLA Me Z 70 a vot. 2102 16 "khm va "ai vet Oh gustu i ba zhude "I syu vari purekkh "azh drink, e Zogurt kum sttyue - tturrUE sashir" REVIVED INJURY tai 1. IN TKA rn ny o i Oko ZAT vah shoi Sure-"tsretekhti zero t Cya Z kudi MI game dodoszhk and skl zsorganizie newly cooked "kash iv Mas ku des l bro sib TV biz Me yishshsh Tve bez Te Ai Ud1 Rich y ; their vka i sh Ko Ko) from Aia Ni Beh siyu Buv BI shi Azhyu Buv tuyu Buv SBedy bej GPU Buv i 5 30 Em o Bbes push ak im iTvo bil Tik b); imml ek Si Ra Bem Avo Aqua ay 35 - so M chi Tush Sh piku st sho j Hr», which OT Eu ru TT piv buh ver chai zr oATa Uai Azrobuch Mai Me buh 10 bo Avro ok BE. 5 BO waza obi av b1u bij Mai Avo vve bity tuyu mmai Uvi Pec di dia to To a po ra an v i i nom nn i I Al bez TV uv vka Su dp oiva Bper rteh Zsr ory Lem Bek y kiv "1 For . "women" BE 135 Vohu taptih 251 SOURCE OF ORIGIN "ida 1. "She" "shchoi Bure" prostheses" "ortachnizme"Woh aty" "bok Mas shu vek siy ii Z1o KhBkoAva Me biv sMH iv Ii dz U Ve 1 Z To ve Nush Ai Ni bej BiUu Buv biu biUu Asia bu Tuye was Gay back ru Pukh 20 aa ZO sii hemo Buv Sich em gbeiy biz otr bib pev osheh Tu Re To Azov 45 Re Terobich two days 5 Zo System division |. SEV value of the immune system! t their beer beer he p; ok, nya s! St | them, and "more. 3 She yi KZ Gu; I V Motto y . i p and field; those yayu CD kh ay: o re, za i in, S fe Y De sr i nok t 4 r y" y - g chk Her. 1. Appearance of antigen and 2. Activation of B-kpitin in it 3. Chemokines, adhesions and activation of T-cells for the production of antibodies to the virus; SEK The autogen-representative cell is the storage of VI Z mys dva a -k . pan and YOU fucking sleep - i vv tsu atak ze "i SM svt. I i NE ST. i iy . she tu "I She, poch ! | i emMOomi me 5 4. Activation of macrophages: 5. Decay of axons, b. Apoptosis of T-KpitiB and devilinization: reduction of trophic functions and ductility of Ma-channels Dysfunction at 4 h meshonduny sh tela at SVO vv" and U . x Ying I -» and Y. ss y 1 « inn y h SM is same AY ba dvy bah o , ha Disintegration of cytoflowers, the first day of occurrence of hpc symptoms: 14 5 z. m te -- a pe and zh sho 10. and sho and | : і х мя ов ше and I І ty 44 | ee her I at SO YANA zbo de Ffn- NN zbo to Kompovkomy Distynsazna. Copaxom and urogeinum B-190 preparation water ZNE-wounds are statistically significant in relation to NND abs. to protein. BO z roi, B - physics are statistically significant in relation to: NND abs, to interferonutamma: z vdo Fig2 Severity of the disease (bai) Animals without clinical signs of the disease during the entire period of the study were not taken into account. « ro ok s Shk a i TU Z YAK a. , Koh 8. yav sk as x. I x pi E X You sh Shchi y; E KE my shi nn F E / ya ry ty g sh Y osyat penya. and KD vn. g / v i NYA fr a. r ps 8 in Zhe them in i teren DD nets MU rub Kt tt tv those Z 3 8 5 vo oloya 8 sho yiv az la 8 boi 8 3 20 31 55 a ba oj zh tv otv Dnuspoterezheniya B NV zbo. to KRN-gamma «yt NND abs. to protein 5-00 o Complex preparation. Copaxone in Distal Bodya x-. satchels are statistically significant in relation to distilled water with ed Fig.Z Severity of the disease (points) Animals without "pinic signs of snoring during the entire period of the study were taken into account I 44 her neck 35 M i i i - p i iii h E ha sha ne noya 2 Z hY Ne y ny y vnu VT ! a NI EMB sp nn i NN drinking par E z: This zhk chakh her o i same z SHENNN 14 23 5 t SOVI ne a SHE i y z 5 34 dk: : VN in Ne. y zha MM U, it zhk Fe. Ж p o ese in v y uh God whining MeV Kk kv m sce Z Yar de: h t SV x5 t EK not i s y: V t m - - Eat daily to drink Days 1-18 | dnz | Days che-3a I GIA ND abs, to protein 3-10 NAD vbe. to 1FN-gamma p g Complex drug EY Copaxone I.I Distidiovana defect with. the differences are statistically significant in relation to distilled water from plants; we. the differences are statistically significant in relation to denatured water from 00 JAN - the differences are statistically significant in relation to NND. or. to the patron 5-100. with ve; 555 - the differences are statistically significant in relation to MND zbo. to isterferonutamma with red du Fig. 4 І "Frequent vinnianennh pecidnyaya | and in, zpzhzhya in do ya renty V uki tteyustnnni VG Y lo i 5 V 1 to. o I dnk nn 2 pemnvnannnnnnnnnnnnnnnnn. t | vro I EZ ek SHI | I osseoya with | tttvevy Va pivu I" | ish and BV OO in B) and pn shi nyny SHY SHE shsh SHE shi and vesha o more I nd to I Zhomplesovy oi Dkstaoivhnya oi opa Fig. 5