RU2505312C2 - Combined therapeutic agent for treating various types of influenza - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: what is administered is a combined therapeutic agent containing an activated potentiated form of human gamma-interferon (IFN-γ) antibodies, an activated potentiated form of CD4 receptor antibodies and an activated potentiated form of histamine antibodies.

EFFECT: effective treatment of the various types of influenza ensured by potentiating the antiviral activity of the activated potentiated gamma-interferon antibodies.

12 cl, 5 tbl, 1 dwg, 4 ex

Description

The invention relates to medicine and can be used to treat influenza, including influenza A and influenza B.

The prior art knows a medicine for the treatment of infectious diseases, including viral etiology, based on the activated form of ultra-low doses of antibodies to interferon (RU 2192888 C1, A61K 39/395, 11/20/2002).

The invention is aimed at improving the treatment of influenza, including influenza A and influenza B.

The solution to this problem is provided by the fact that a comprehensive drug for the treatment of various types of influenza, containing the activated-potentiated form of antibodies to human gamma-interferon, according to the invention, contains, as additional active components, the activated-potentiated form of antibodies to CD4 and the activated-potentiated form of antibodies to histamine.

In this case, the activated-potentiated form of antibodies to human gamma-interferon (IFN-γ), the activated-potentiated form of antibodies to the CD4 receptor and the activated-potentiated form of antibodies to histamine are used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution, the activity of which is due to the process of sequential multiple dilution of the matrix initial solution of antibodies in an aqueous or aqueous-alcohol solvent in combination with an external mechanical action - vertical by shaking of each dilution, with final dilutions contain ultralow doses (SMD) antibodies.

In this case, the complex drug can be made in solid dosage form in the form of a pharmaceutical composition and contain the technologically necessary (effective) amount of a neutral carrier saturated with a mixture of the activated-potentiated form of antibodies to human gamma-interferon, the activated-potentiated form of antibodies to the CD4 receptor and activated -potentiated forms of antibodies to histamine, and pharmaceutically acceptable additives, which include, for example, lactose, microcrystalline cellulose th and magnesium stearate.

In this case, aqueous or aqueous-alcoholic solutions of activated - potentiated forms of antibodies to human gamma interferon, to the CD4 receptor and to histamine are obtained by repeated serial dilution of the matrix-initial solution of antibodies to human gamma interferon, to the CD4 receptor and to histamine in combination with external influence - vertical shaking of each dilution, while the concentration of matrix solutions is 0.5 ÷ 5.0 mg / ml.

Preferably, each of the components of the claimed medicinal product is used as a mixture of various, mainly hundred, dilutions prepared by homeopathic technology.

The solution to this problem is also provided by the fact that in the method of treating various types of influenza by introducing into the body an activated - potentiated form of antibodies to human gamma-interferon, according to the invention, an activated (potentiated) form of antibodies to the CD4 receptor and an activated potentiated form of antibodies to histamine.

Preferably, the activated is the potentiated form of antibodies to human gamma interferon, the activated potentiated form of antibodies to the CD4 receptor, and the activated

- the potentiated form of antibodies to histamine is used in the form of an activated-potentiated aqueous or aqueous-alcoholic solution of each component, the activity of which is due to the process of successive multiple dilutions of the matrix-initial-solution of antibodies in an aqueous or aqueous-alcoholic solvent in combination with an external mechanical action - vertical shaking each breeding.

Moreover, the indicated influenza virus can be, for example, influenza A, including seasonal flu, H5N1 and H1N1, and influenza B.

At the same time, they can use a mixture of different dilutions of antibodies to human gamma-interferon prepared in the form of a single drug - a single dosage form in combination (in combination) with a mixture of different dilutions of antibodies to the CD4 receptor and to histamine prepared using homeopathic technology.

At the same time, each of the active ingredients is a mixture of hundreds of dilutions prepared according to homeopathic technology.

In addition, the claimed drug, as complex, contains active ingredients in a volume 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 ⁵⁰ times, which is equivalent hundred dilutions of C12, C30, C50, prepared according to homeopathic technology.

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

In the treatment of various types of influenza, including influenza A, including seasonal flu, H5N1 and H1N1, and influenza B, 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 the activated-potentiated form of antibodies to human gamma-interferon, the activated-potentiated form of antibodies to the CD4 receptor and, accordingly, the activated-potentiated form of antibodies to histamine.

According to the invention, the activated-potentiated form of antibodies is an antibody form, for example, to human gamma-interferon, to a CD4 receptor or to histamine, prepared according to the homeopathic potentiation technique by repeated serial dilution of the matrix-source antibody solution in combination with an external action - vertical shaking each dilution that is active in pharmacological models and / or clinical treatments for various types of influenza, including high atogenic flu, influenza A, including seasonal flu, H5N1 and H1N1, and influenza B.

The proposed combination of activated-potentiated forms of antibodies to human gamma-interferon, to CD4 receptor and to histamine provides an unexpected synergistic therapeutic effect, experimentally confirmed in an adequate model, which consists in increasing the effectiveness of treatment of various types of influenza, including highly pathogenic influenza, influenza A, in including seasonal flu, H5N1 and H1N1, and influenza B, which is due to increased antiviral activity and increased immunotropic effects due to increased fun tional activity of the CD4 receptor, receptor gamma - interferon and histamine. In addition, the use of the claimed drug in the pre- and post-vaccination period increases the effectiveness of vaccination, provides non-specific prophylaxis of acute respiratory viral infections and influenza at the time of establishment of post-vaccination immunity, prevents the development of intercurrent diseases in the post-vaccination period.

In this case, it is possible to use the claimed complex drug in combination with other antiviral drugs and symptomatic agents.

A drug is prepared mainly as follows.

To prepare the activated-potentiated form of the active components, monoclonal or, mainly, polyclonal antibodies are used, which can be obtained according to well-known technology-methods described, for example, in the book: Immunological methods, ed. G. Frimel, 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 biotechnological 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 hybridoma cells producing clones of antibodies of the same specificity. Their isolation in an individual form is carried out by the same methods as in the case of polyclonal antisera.

Polyclonal antibodies can be obtained by active immunization of animals. For this purpose, according to a specially developed scheme, the animals are given a series of injections of the substance-antigen or conjugated antigen required by the invention: human gamma-interferon, CD4 receptor and histamine. 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.

Example 1

To obtain polyclonal antibodies to human gamma-interferon, an adjuvant and, for example, a whole human gamma-interferon molecule of the following sequence are used as immunogen (antigen) for immunization of rabbits:

1 MKYTSYILAF QLCIVLGSLG CYCQDPYVKE AENLKKYFNA GHSDVADNGT LFLGILKNWK

61 EESDRKIMQS QIVSFYFKLF KNFKDDQSIQ KSVETIKEDM

NVKFFNSNKK KRDDFEKLTN

121 YSVTDLNVQR KAIHELIQVM AELSPAAKTG KRKRSQMLFR

GRRASQ

It is possible to obtain a polyclonal antibody to human gamma interferon as an immunogen (antigen) for immunization of rabbits using an adjuvant and, for example, one polypeptide fragment of human gamma interferon selected from the following sequences:

7-55:

ILAF QLCIVLGSLG CYCQDPYVKE AENLKKYFNA GHSDV

24-166:

QDPYVKEAE NLKKYFNAGH SDVADNGTLF LGILKNWKEE

SDRKIMQSQI VSFYFKLFKN FKDDQSIQKS VETIKEDMNV

KFFNSNKKKR DDFEKLTNYS VTDLNVQRKAIHELIQVMAE

LSPAAKTGKR KRSQMLFRGR RASQ

24-166:

QDPYVKEAE NLKKYFNAGH SDVADNGTLF LGILKNWKEE

SDRKIMQSQI VSFYFKLFKN FKDDQSIQKS VETIKEDMNV

KFFNSNKKKR DDFEKLTNYS VTDLNVQRKA IHELIQVMAE

LSPAAKTGKR KRSQMLFQGR RASQ

69-123:

QS QIVSFYFKLF KNFKDDQSIQ KSVETIKEDM NVKFFNSNKK

KRDDFEKLTN YSV

100-145:

M NVKFFNSNKK KRDDFEKLTN YSVTDLNVQR KAIHELIQVM

AELSP

92-130:

SVETIKEDM NVKFFNSNKK KRDDFEKLTN YSVTDLNVQR

123-147:

VTDLNVQR KAIHELIQVM AELSPAA

24-166:

MQDPYVKEAE NLKKYFNAGH SDVADNGTLF LGILKNWKEE

SDRKIMQSQI VSFYFKLFKN FKDDQSIQKS VETIKEDMNV

KFFNSNKKKR DDFEKXTNYS VTDLNVQRKAIHELIQVMAE

LSPAAKTGKR KRSQMLFQGR RASQ

5-45:

SYILAF QLCIVLGSLG CYCQDPYVKE AENLK

94-114:

ETIKEDM NVKFFNSNKK KRDD,

24-166:

MQDPYVKEAE NLKKYFNAGH SDVADNGTLF LGILKNWKEE

SDRKIMQSQI VSFYFKLFKN FKDDQSIQKS VETIKEDMNV

KFFNSNKKKR DDFEKXTNYS VTDLNVQRKA IHELIQVMAE

LSPAAKTGKR KRSQMLFRGR RASQ

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 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. You can add 20% NaN ₃ to the antiserum 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 °). The selection of anti-gamma interferon antibodies from the antiserum is possible as follows:

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.

Antibodies are purified by affinity chromatography on a column with antigen by binding of antibodies to gamma interferon with an antigen (gamma interferon) attached to an insoluble matrix of the column, followed by elution of the antibodies with concentrated salt solutions.

Thus obtained buffer solution of polyclonal rabbit anti-gamma interferon antibodies purified on an antigen with a concentration of 0.5 ÷ 5.0 mg / ml, preferably 2.0 ÷ 3.0 mg / ml, is used as a matrix (initial ) a solution for the subsequent preparation of the activated-potentiated form.

Polyclonal antibodies to the CD4 receptor are obtained according to the above method, using an immunogen (antigen) for immunization of rabbits with an adjuvant and a whole molecule or one polypeptide fragment of the CD4 receptor, selected, for example, from the following amino acid sequences:

412-458:

IGLGIFFCV RCRHRRRQAE RMSQIKRLLS EKKTCQCPHR

FQKTCSPI

26-60:

MNRGVPFRHL LLVLQLALLP AATQGKKVVL GKKGD

105-119:

A DSRRSLWDQG NFPL

115-139:

G NFPLIIKNLKIEDSDTYICE VEDQ

26-458: MNRGVPFRHL LLVLQLALLP AATQGKKVVL GKKGDTVELT CTASQKKSIQ FHWKNSNQIK ILGNQGSFLT KGPSKLNDRA DSRRSLWDQG NFPLIIKNLK IEDSDTYICE VEDQKEEVQL LVFGLTANSD THLLQGQSLT LTLESPPGSS PSVQCRSPRG KMQGGKTLS VSQLELQDSG TWTCTVLQNQ KKVEFKIDIV VLAFQKASSI VYKKEGEQVE FSFPLAFTVE KLTGSGELWW QAERASSSKS WITFDLKNKE VSVKRVTQDP KLQMGKKLPL HLTLPQALPQ YAGSGNLTLA LEAKTGKLHQ EVNLVVMRAT QLQKNLTCEV WGPTSPKLML SLKLENKEAK VSKREKAVWV LNPEAGMWQC LLSDSGQVLL ESNIKVLPTW STPVQPMALI VLGGVAGLLL FIGLGIFFCV RCRHRRRQAE RMSQIKRLLS EKKTCQCPHR FQKTCSPI

The preparation of polyclonal antibodies to histamine can be carried out according to the above method for the preparation of polyclonal antibodies to human gamma interferon using the immunogen (antigen) for immunization of rabbits with adjuvant and histamine (4- (2-aminoethyl) -imidazole, or beta-imidazolyl-ethylamine with the chemical formula C ₅ H ₉ N ₃ .

Preferred for the preparation of the claimed drug is the use of polyclonal antibodies to human gamma-interferon, to CD4 (for example, to CD4 receptor of T-lymphocytes) and to histamine, which as a matrix (initial) solution with a concentration of 0.5 ÷ 5.0 mg / ml is used for 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 successive dilution of 1 part of each solution to be diluted, starting from the above matrix solution, in 9 parts (for decimal dilution D) or 99 parts (for hundredth dilution C) or in 999 parts (for a thousandth dilution of M) of a neutral solvent in combination with multiple vertical shaking (potentiation, or "dynamization") of each dilution obtained and using tdelnyh containers for each subsequent dilution to the required potency - the multiplicity of homeopathic dilution method (see, eg, W. Schwabe "Homeopathic medicines", M., 1967, s.14-29.).

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

For example, to prepare the 12th hundredth C12 dilution, one part of a matrix (initial) antibody solution, for example, to human gamma-interferon, with a concentration of 2.5 mg / ml, is diluted in 99 parts of a neutral aqueous or aqueous-alcoholic solvent and repeatedly (10 and more than once) vertically shake-potentiate, getting the 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 C12 is a solution obtained by diluting 12 times in succession in different containers of the first part of the initial matrix solution of antibodies to human gamma-interferon with a concentration of 2.5 mg / ml in 99 parts of a neutral solvent, i.e. the solution obtained by diluting the matrix solution in 100 ¹² times. Similar operations with the appropriate dilution ratio are carried out to obtain dilutions of C30 and C50.

When using, for example, the activated-potentiated form of antibodies to human gamma-interferon in the form of a mixture of various, mainly hundred, dilutions, each dilution (for example, C12, C30, C50) is prepared separately according to the above technology until the dilution is 3 dilutions less than the final (respectively, to obtain C9, C27, C47), and then make, in accordance with the composition of the mixture, one part of each component in one container and mixed with the required amount of solvent (respectively, with 97 parts for a hundredth dilution ny). Next, the resulting mixture was successively diluted twice in a ratio of 1 to 100, potentiating the resulting solution after each dilution. In this case, an activated - potentiated form of antibodies is obtained, for example, to human gamma-interferon in an ultra-low dose obtained by diluting the matrix solution 100 ¹² , 100 ³⁰ , 100 ⁵⁰ times, equivalent to a mixture of hundred-percent dilutions C12, C30, C50.

It is possible to use each component in the form of a mixture of other different dilutions, for example, decimal and or hundredths (D20, C30, C100 or C12, C30, C200, etc.) prepared according to homeopathic technology, the effectiveness of which is determined experimentally.

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

The claimed drug can also be made in solid dosage form in the form of a pharmaceutical composition that contains a technologically necessary (effective) amount of a neutral carrier — for example, lactose — saturated by soaking in a mixture of aqueous or aqueous-alcoholic solutions of an activated-potentiated form of gamma antibodies to human interferon, an activated-potentiated form of antibodies to the CD4 receptor and an activated-potentiated form of antibodies to histamine, and a pharmaceutical and 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, Huttlin Pilotlab type manufactured by Htittlin GmbH) irrigation until saturation of granules of a neutral substance - lactose (milk sugar) with a particle size of 50 ÷ 500 microns, pre-prepared aqueous or aqueous-alcoholic solution of an activated potentiated form of antibodies to the CD4 receptor, mainly in the ratio of 1 kg of antibody solution to 5 or 10 kg of lactose (1: 5-1: 10) with simultaneous drying a second flow of heated air supplied under the grate at a temperature of not higher than 40 ° C. The estimated amount of lactose (10 ÷ 91% of the tablet mass) saturated with the activated-potentiated form of antibodies according to the above procedure is loaded into the mixer and mixed with lactose, moistened with the activated-potentiated form of antibodies, in an amount of 3 ÷ 10% of the tablet mass and with pure lactose in an amount of not more than 84% of the tablet mass (to reduce the cost and some simplification and acceleration of the process without reducing the effectiveness of the therapeutic effect). Then, microcrystalline cellulose is added to this mixture in an amount of 5-10% of the tablet mass and magnesium stearate in an amount of 1% of the tablet mass. 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 with an activated-potentiated form of antibodies to human gamma interferon, to CD4 T-lymphocyte receptor and to histamine in an ultra-low dose of each component prepared from a matrix solution diluted in 10012, in 10030 in 10050, which is equivalent to a mixture of hundred-percent dilutions of C12, C30 and C50 prepared according to homeopathic technology.

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

Example 2

To study the properties of the claimed drugs for the treatment of patients of the active drug group No. 1, 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 gamma interferon (anti-IFN-γ) and CD4 receptor (anti-CD4) at ultra low doses (RMA) obtained sverhrazvedeniem initial matrix solution 100 ^12, 100 ^30, 100 ^{to 200} times equivalent mixture of centesimal gomeopatiche FIR dilutions C12, C300, C200 (SMD anti-IFN-γ + anti-CD4); for the treatment of patients of the active drug group No. 2, 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 gamma interferon (anti-IFN-γ ), to the CD4 receptor (anti-CD4) and histamine (anti-H) in ultra-low doses (SMD) obtained by super-dilution of the initial matrix solution 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times, equivalent to a mixture of hundred-percent homeopathic dilutions C12, C30, C200 (SMD anti-IFN-γ + STI-CD4 + anti-H). 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 human gamma interferon, purified on antigen, in an ultra-low dose obtained by super-dilution of the initial matrix solution in 100 ¹² , 100 ³⁰ , 100 ⁵⁰ times, the equivalent mixture of hundreds of homeopathic dilutions C12, C30, C50 (SMD anti-IFN-γ).

The antiretroviral activity of the SMD anti-IFN-γ + anti-CD4 and SMD anti-IFN-γ + anti-CD4 + anti-H compositions was evaluated in an open comparative clinical trial in patients infected with human immunodeficiency virus (HIV), which were observed in Territorial Center for the Prevention and Control of AIDS and Infectious Diseases. The study included 65 men and 32 women (a total of 97 patients) aged 18 to 48 years with a viral load of ≥150 copies of HIV-1 RNA in 1 ml of plasma and a CD4-lymphocyte level of at least 250 cells / μl (or ≥0 , 25 × 10 ⁹ / L). 34 of the 97 study participants were observed for asymptomatic infectious status and had not previously received treatment for HIV infection (naive patients). 63 of 97 patients were on antiretroviral therapy (APT) for 1-2 years. The study did not include patients with liver cirrhosis, viral hepatitis C, severe concomitant diseases in the acute stage, pregnant women, as well as persons taking intravenous drugs. The study was conducted in the autumn-winter period, when there was a seasonal increase in the incidence of influenza and acute respiratory infections (ARVI).

75 patients were randomized into three groups, they were prescribed therapy with group 1 and 2 drugs or group 3 comparison drug in a prophylactic (against influenza / SARS) dosage - 1 tablet 1 time per day for 6 weeks, including: patients group 1 (n = 25) was assigned the composition of SMD anti-IFN-γ + anti-CD4 in the form of monotherapy (subgroup 1A: naive patients, n = 12) or in combination with APT (subgroup 1B, n = 13); group 2 patients (n = 23) were prescribed SMD anti-IFN-γ + anti-CD4 + anti-H compositions, including monotherapy (subgroup 2A: naive patients, n = 11) or in combination with APT ( subgroup 2B, n = 12); group 3 patients (n = 27) were prescribed SMD anti-IFN-γ, including monotherapy (subgroup 3A: naive patients, n = 11) or in combination with APT (subgroup 3B, n = 16). The control (fourth) group (n = 22) consisted of patients who continued to receive APT according to the previous scheme without adding any study drug (APT group).

At baseline and after 6 weeks, all patients were assessed for viral load, CD4 and CD8 lymphocyte counts, and CD4 / CD8 immunoregulatory index. To determine the copies of HIV-1 RNA in blood plasma, we used the COBAS AMPLICOR HTV-1 MONITOR Kit, version 1.5 for the COVAS AMPLICOR automatic PNR analyzer (Roche, Switzerland). Phenotyping of circulating peripheral blood lymphocytes was performed on a FACSCount flow cytometer (Becton Dickinson, USA) using standard FACSCount Reagent Kit test systems containing antibodies to CD3, CD4, CD8 labeled with FITC, PE fluorochromes.

Table 1 presents data on the viral load (the number of copies of HIV RNA) in the form of the median (Me), first and third quartiles (Q1, Q3) in patients of the studied groups in the dynamics of observation. As a result of the study, it was found that a 6-week intake of the SMD anti-IFN-γ + anti-CD4 composition reduced the number of copies of HIV-1 RNA in 58% of naive patients (in 7 of 12 people of subgroup 1A), while the average decrease in viral the load was 16.9%. The combined use of the composition of SMD anti-IFN-γ + anti-CD4 and APT was no less effective, since a decrease in the number of copies of HIV-1 RNA was observed in 62% of the study participants (in 8 out of 13 people of subgroup 1B), and the average viral load reduction was 18.2% of the baseline. Approximately similar indicators were obtained using the composition of SMD anti-IFN-γ + anti-CD4 + anti-H: antiviral activity was recorded in 55% of HIV-infected naive patients (in 6 of 11 people of subgroup 2A) and in 67% of people who received the combination SMD anti-IFN-γ + anti-CD4 + anti-H and APT (in 8 of 12 people of subgroup 2B); the average decrease in viral load was 17.3%) and 18.9%, respectively. The antiretroviral activity observed in the first two groups was slightly higher than the treatment results obtained in patients of the comparison group. After 6 weeks, monotherapy with SMD anti-IFN-γ reduced the number of copies of HIV-1 RNA in 36% of naive patients (in 4 of 11 people of subgroup 3A), while the decrease in viral load averaged 9.5%. The combined use of the monocomponent drug SMD anti-IFN-γ and APT increased the effectiveness of therapy, helping to reduce the viral load in 50% of the study participants (in 8 of 16 people of subgroup 3B) by an average of 14.2%. Similar treatment indices in individuals receiving only APT (control group 4) were 32% (in 7 of 22 people in group 4) and 13.3%, respectively.

Analysis of the indicators of the subpopulations of circulating lymphocytes in the dynamics of observation (Table 2) showed a more pronounced increase in the number of CD4 lymphocytes than in the control group after 6 weeks of therapy using SMD anti-IFN-γ + anti-CD4, SMD anti-IFN-γ + anti -CD4 + anti-H and SMD anti-IFN-γ as monotherapy in naive patients (subgroups 1A, 2A and 3A) or in combination with APT (subgroups 1B, 2B and 3B). Moreover, the number of CD8 after 6 weeks of drug administration (mono-or combination therapy) did not change in any of the studied groups. The positive dynamics of SB4 lymphocytes during treatment led to an increase in the immunoregulatory index of CD4 / CD8, the most significant in the subgroups of patients taking SMD compositions anti-IFN-γ + anti-CD4 and anti-IFN-γ + anti-CD4 + anti H (mono - or combination therapy - groups I and 2) and SMD anti-IFN-γ in combination with APT (subgroup 3B).

Table 1 Dynamics of viral load depending on the treatment regimen Viral load, copies / ml The average decrease in viral load,% SMD anti-IFN-γ + anti-CD4 (Me [Q1-Q3]) Originally 5769 [368-62584] 16.9 After 6 weeks of treatment 4575 [337-58526] APT and SMD anti-IFN-γ + anti-CD4 (Me [Q1-Q3]) Originally 5238 [385-59695] 18.2 After 6 weeks of treatment 4408 [320-50197] SMD anti-IFN-γ + anti-CD4 + anti-H (Me [Q1-Q3]) Originally 5638 [385-61742] 17.3 After 6 weeks of treatment 4754 [278-57426] APT and SMD anti-IFN-γ + anti-CD4 + anti-H (Me [Q1-Q3]) Originally 5189 [350-59798] 18.9 After 6 weeks of treatment 46108 [269-47987] SMD anti-IFN-γ (Me [Q1-Q3]) Originally 5813 [150-33356] 9.5 After 6 weeks of treatment 5786 [150-38359] APT and SMD anti-IFN-γ (Me [Q1-Q3]) Originally 4680 [274-9838] 14.2 After 6 weeks of treatment 4652 [272-8874] APT (Me [Q1-Q3]) Originally 5547 [385-58996] 13.3 After 6 weeks of treatment 5308 [338-57709]

table 2 Indicators of subpopulations of circulating lymphocytes in patients of the studied groups in the dynamics of observation Observation period CD4, cells / μl (M ± SE) CD4 / CD8 (M ± SE) SMD anti-IFN-γ + anti-CD4 (n = 12) Originally 516 ± 33 0.46 ± 0.09 After 6 weeks of treatment 712 ± 24 0.58 ± 0.07 * APT and SMD anti-IFN-γ + anti-CD4 (n = 13) Originally 499 ± 41 0.50 ± 0.08 After 6 weeks of treatment 728 ± 29 0.60 ± 0.06 * SMD anti-IFN-γ + anti-CD4 + anti-H (n = 11) Originally 509 ± 45 0.49 ± 0.06 After 6 weeks of treatment 706 ± 27 0.58 ± 0.08 * APT and SMD anti-IFN-γ + anti-CD4 + anti-H (n = 12) Originally 521 ± 37 0.48 ± 0.09 After 6 weeks of treatment 734 ± 22 0.62 ± 0.10 * SMD anti-IFN-γ (n = 11) Originally 513 ± 98 0.38 ± 0.19 After 6 weeks of treatment 563 ± 26 0.44 ± 0.12 APT and SMD anti-IFN-γ (n = 16) Originally 491 ± 49 0.55 ± 0.06 After 6 weeks 623 ± 45 0.67 ± 0.05 * treatment APT (n = 22) Originally 510 ± 29 0.44 ± 0.06 After 6 weeks of treatment 595 ± 35 0.50 ± 0.12 * significant differences compared to baseline, p <0.05

The absence in patients during treatment of adverse events associated with the administration of the studied drugs indicated a good tolerance to the drugs. Repeated monitoring of laboratory parameters, including blood and urine tests, biochemical markers, confirmed the safety of treatment.

Thus, the study demonstrated the antiretroviral efficacy of the drug SMD anti-IFN-γ + anti-CD4 and the drug SMD anti-IFN-γ + anti-CD4 + anti-H, apparently due to a change in the functional activity of CD4 receptors, which prevents the penetration of HIV into cells, as well as suppression of HIV replication inside the cell due to activation of transcription processes of antiviral protein mRNA. It was found that the decrease in viral load observed as a result of a 6-week intake of the drug SMD anti-IFN-γ + anti-CD4 and the drug SMD anti-IFN-γ + anti-CD4 + anti-H at a dose of 1 tablet per day exceeds the antiviral the effect that is recorded after 6 weeks of taking the monocomponent drug SMD anti-IFN-γ in the same dose or continuing according to the previous APT regimen. The combination of any drug SMD aHTH-IFN-γ + aHTH-CD4, SMD anti-IFN-γ + anti-CD4 + anti-H or SMD anti-IFN with APT slightly enhances the antiviral activity of the latter, which is manifested in a decrease in the average value of viral load after 6 weeks in a larger percentage of study participants.

The effect of SMD preparations anti-IFN-γ + anti-CD4 and anti-IFN-γ + anti-CD4 + anti-H on the ratio of CD4 / CD8 lymphocytes in HIV-infected patients (due to a slight increase in the absolute number of CD4 cells) was shown. most pronounced when used with APT. Given the simultaneous decrease in viral load in patients taking SMD anti-IFN-γ + anti-CD4 and SMD anti-IFN-γ + anti-CD4 + anti-H, it can be assumed that the increase in the number of CD4 cells is associated with an increase in the population of these cells due to healthy (uninfected) cells. The combination of APT with SMD anti-IFN-γ + anti-CD4, SMD anti-IFN-γ + anti-CD4 + anti-H or SMD anti-IFN is more effective in restoring the immunoregulatory index of CD4 / CD8 than the ongoing APT as before scheme.

The antiretroviral activity of the drugs SMD anti-IFN-γ + anti-CD4 and SMD anti-IFN-γ + anti-CD4 + anti-H allows them to be used for prophylactic and therapeutic purposes both in naive HIV-infected patients and in patients receiving Apt.

Example 3

For experimental study of the claimed technical solution, 300 mg tablets were used, impregnated with a pharmaceutical composition containing water-alcohol solutions (6 mg / tablet) of activated - potentiated forms of polyclonal affinity purified rabbit antibodies to human gamma interferon (AT GI), to the CD4 receptor molecule of T-lymphocytes (AT CD4), to histamine (AT G), in ultra-low doses (SMD) obtained by super-dilution of the initial matrix solution in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times of each component, equivalent to a mixture of hundred homeo pathological dilutions C12, C30, C200 (AT GI + AT CD4 + AT G). As the reference drug using the drug containing activated - potentiated form of polyclonal affinity-purified rabbit antibody to gamma interferon, human (AT GOP) in ultralow doses (RMA) obtained sverhrazvedeniem initial matrix solution 100 ^12, 100 ^30, 100 ^{to 200} times equivalent mixture hundredth homeopathic dilutions C12, C30, C200 (SMD AT GI). The placebo group received 300 mg tablets containing lactose monohydrate, microcrystalline cellulose, magnesium stearate.

In the ongoing double-blind, placebo-controlled study of the efficacy and safety of the drug AT GI + AT CD4 + AT G in SARS and in the ongoing open comparative controlled study of the efficacy and safety of the drug AT GI + AT CD4 + AT G in influenza, the number of people developing against the background of an acute infectious process was additionally evaluated complications, including bacterial (bacterial pneumonia, tracheitis, otitis media, glomerulonephritis, etc.).

In cases where the defense system is perfect, the infectious process may be interrupted or, remaining localized, not accompanied by the development of severe clinical symptoms, i.e. the adequacy of protective reactions leads to rapid inactivation of the pathogen, the restoration of impaired body functions and recovery. A different picture arises in an organism highly susceptible to an infectious agent and lacking the perfect mechanism of specific and nonspecific protection (immunocompromised patients). In such cases, pathogens reproduced in an increasing number and the products of their interaction with epithelial and immune cells, as well as the destroyed cells themselves enter the blood, causing the development of severe forms of the course of the disease, the formation of complications and a possible adverse outcome.

The use of the drug AT GI + AT CD4 + AT G both in case of influenza and ARVI significantly reduces the frequency of bacterial complications compared with placebo (Table 3), and, as a result, reduces the need for antibiotic therapy. The drug AT GI + AT CD4 + AT G prevents the development of secondary immunodeficiency at the stage of convalescence, providing an immunomodulating effect and increasing the body's natural defenses. The ability of AT GI + AT CD4 + AT G to reduce the incidence of bacterial complications was greater than that of SMI AT GI.

Table 3 The incidence of bacterial complications A drug Patient count Bacterial complications Otitis count /% Tracheitis number /% Pneumonia qty /% Total Qty /% AT GI + AT CD4 + AT G (ARVI) 40 0/0 1 / 2.5 0/0 1 / 2.5 Placebo (ARVI) 38 3 / 7.9 7 / 18.4 0/0 10 / 26.3 AT GI + AT CD4 + AT G (Flu) 6 0/0 0/0 0/0 0/0 Tamiflu® (Flu) eleven 0/0 2 / 18.2 1/9.1 3 / 27.2 SMD AT GI (Influenza and SARS) * thirty 1 / 3.3 2 / 6.7 0/0 3 / 10.0 Placebo (Flu & ARVI) * thirty 4 / 13.3 5 / 16.7 0/0 9/30 * - According to a double-blind, placebo-controlled randomized study of the clinical efficacy and safety of the therapeutic use of the drug SMD AT GI for influenza and other acute respiratory viral infections (GU Research Institute of Influenza RAMS, St. Petersburg, 2005).

Example 4

For experimental study of the claimed technical solution, 300 mg tablets were used, impregnated with a pharmaceutical composition containing water-alcohol solutions (6 mg / tablet) of activated - potentiated forms of polyclonal affinity purified rabbit antibodies to human gamma interferon (AT GI), to the CD4 receptor molecule of T-lymphocytes (AT CD4), to histamine (AT G), in ultra-low doses (SMD) obtained by super-dilution of the initial matrix solution in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ times of each component, equivalent to a mixture of hundred homeo pathological dilutions C12, SZO, C200 (AT GI + AT CD4 + AT G). Tamiflu® (the active ingredient is oseltamivir, F. Hoffmann-La Roche Ltd. - Switzerland) (Oseltamivir) was used as a comparison drug.

The ongoing open comparative controlled clinical trial of AT GI + AT CD4 + AT G and Tamiflu® includes patients aged 18-60 years of both sexes, with influenza A or B with signs of intoxication and catarrhal syndrome. The study includes patients with a body temperature of 37.8 ° C or more (provided that such a temperature is recorded from the moment of the onset of the disease), the duration of the disease is not more than 48 hours before treatment, without severe concomitant diseases. Patients undergo a rapid test for the presence of influenza antigens. Patients with negative test results are not included in the study. All patients give written voluntary consent to participate in the study before starting all study procedures. Patients fill out diaries in which they record body temperature twice a day, concomitant therapy, etc. Patients receive AT GI + AT CD4 + AT G, 8 tablets on the first day and three tablets a day from the second to the fifth days, or Tamiflu® 75 mg twice daily, according to manufacturer's instructions. Patients can take antipyretic drugs if necessary. The use of antiviral, immunomodulatory, antihistamines and antibiotics is not allowed. Before the first dose and at the last visit, patients give blood and urine samples for laboratory tests in order to monitor the safety of the therapy. Treatment lasts 5 days, follow-up - 2 days. Thus, on average, each patient takes part in the study within 7 days.

The effectiveness of therapy was evaluated by the timing of reaching body temperature of 37.0 ° C or less, the number of antipyretic doses was also compared.

At the time of the analysis, the study completed 17 patients (6 patients in the AT GI + AT CD4 + AT G group and 11 patients in the Tamiflu® group).

The proportion of patients with a body temperature of 37.0 ° C or less in the groups did not statistically differ among themselves throughout the treatment period. By the beginning of 4 days of treatment in both groups was achieved almost complete recovery of patients (Figure 2). By the beginning of the second day of treatment in both groups, a third of patients had normalized body temperature. The average number of antipyretic doses in the groups also did not have statistically significant differences and at the beginning of the 4th day of therapy was 7.6 ± 0.84 in the AT GI + AT CD4 + AT G group and 7.4 ± 0.90 in the group taking Tamiflu®, respectively.

The safety analysis included data from all 17 patients participating in the study and completing treatment within the protocol established terms; there were no early drop-out patients. During the entire observation period, the drug was well tolerated, adverse events were not observed. A blood test at the beginning and end of the treatment period did not reveal pathological abnormalities. A general urinalysis on the first and final day of the study also did not reveal any pathology in any patient.

When comparing the data with the results of a 2005 double-blind, placebo-controlled, randomized study of the clinical efficacy and safety of the therapeutic use of a drug containing activated - potentiated forms of polyclonal affinity purified rabbit antibodies to human gamma interferon (ATG) in ultra-low doses (SMD) obtained by ultra-dilution initial matrix solution 100 ^12, 100 ^30, 100 ^{to 200} times equivalent mixture of centesimal homeopathic dilutions C12, C30, C200 (SMD aT HY) In case of influenza and other acute respiratory viral infections (State Research Institute of Influenza, Russian Academy of Medical Sciences, St. Petersburg, 2005), it was found that AT GI + AT CD4 + AT G is more effective than AT SM GI in lowering the temperature (see Figure 1, Table 4 and Table 5).

Claims (12)

1. A comprehensive drug for the treatment of various types of influenza, characterized in that it contains an activated - potentiated form of antibodies to human gamma-interferon, an activated - potentiated form of antibodies to a CD4 receptor and an activated - potentiated form of antibodies to histamine. 2. The complex drug according to claim 1, characterized in that the activated — potentiated form of antibodies to human gamma-interferon, the activated — potentiated form of antibodies to the CD4 receptor, and the activated — potentiated form of antibodies to histamine are used in the form of an activated — potentiated water or water -alcohol solution, the activity of which is due to the process of successive multiple dilutions of the matrix - initial - solution of antibodies in an aqueous or aqueous-alcoholic solvent in the blood with external mechanical action - vertical shaking of each dilution. 3. The complex drug according to claim 1, characterized in that it is made in solid dosage form in the form of a pharmaceutical composition and contains the technologically necessary amount of a neutral carrier saturated with a mixture of activated potentiated forms of antibodies to human gamma-interferon, activated - potentiated forms of antibodies to CD4 receptor and activated - potentiated forms of antibodies to histamine, and pharmaceutically acceptable additives. 4. The complex drug according to claim 1, characterized in that aqueous or aqueous-alcoholic solutions of activated - potentiated forms of antibodies to human gamma interferon, to the CD4 receptor and to histamine are obtained by repeated serial dilution of matrix solutions of antibodies to human gamma interferon , to the CD4 receptor and to histamine in combination with an external action - vertical shaking of each dilution, while the concentration of the matrix solutions is 0.5 ÷ 5.0 mg / ml. 5. The complex drug according to claim 1, characterized in that each of the components is used in the form of a mixture of various, mainly hundred, dilutions prepared by homeopathic technology. 6. The complex drug according to claim 3, characterized in that the pharmaceutically acceptable additives include lactose, microcrystalline cellulose and magnesium stearate. 7. A method for treating various types of influenza by introducing into the body an activated - potentiated form of antibodies to human gamma-interferon, characterized in that an activated - potentiated form of antibodies to the CD4 receptor and an activated - potentiated form of antibodies to histamine are additionally simultaneously and simultaneously administered. 8. The method according to claim 7, characterized in that the activated — potentiated form of antibodies to human gamma-interferon, the activated — potentiated form of antibodies to the CD4 receptor, and the activated — potentiated form of antibodies to histamine, are used in the form of an activated — potentiated water or water-alcohol a solution of each component, the activity of which is due to the process of successive multiple dilutions of the matrix - initial - solution of antibodies in an aqueous or aqueous-alcoholic solvent in combination with another mechanical action - vertical shaking of each dilution. 9. The method according to claim 7, characterized in that said type of flu is influenza A or influenza B. 10. The method according to claim 9, characterized in that the specified influenza A is influenza A strain H5N1 and H1N1. 11. The treatment method according to claim 7, characterized in that a mixture of different dilutions of antibodies to human gamma-interferon in combination with a mixture of different dilutions of antibodies to the CD4 receptor and histamine prepared according to homeopathic prepared in the form of a single drug is used technologies. 12. A pharmaceutical composition for treating various types of influenza, characterized in that it contains a mixture of the activated - potentiated form of antibodies to human gamma interferon, the activated - potentiated form of antibodies to the CD4 receptor and the activated - potentiated form of antibodies to histamine.

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