RU2525156C2 - Method of treatment and prevention of arterial hypertension and pharmaceutical composition for treatment of arterial hypertension - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed group of inventions relates to medicine, namely to cardiology, and deals with treatment and prevention of arterial hypertension. For this purpose pharmaceutical composition, containing activated potentiated antibodies to angiotensin II receptor and to endothelial NO-synthase, is introduced.

EFFECT: method ensures effective treatment of arterial hypertension due to synergic antihypertensive action of composition components.

11 cl, 2 ex, 1 tbl

Description

The invention relates to medicine and can be used to increase the effectiveness of the treatment of arterial hypertension of the I-II degree, as well as primary and secondary prevention of arterial hypertension.

The prior art preparation for the treatment of arterial hypertension based on activated potentiated forms of ultra-low doses of antibodies to the angiotensin II receptor (Treatment of hypertension using ultra-low doses of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor. Petrov V.I. et al. Arterial hypertension, 2008, p. 236-238). However, this drug does not in all cases provide sufficient therapeutic efficacy.

The invention is aimed at increasing the effectiveness of the treatment of arterial hypertension of the I-II degree, as well as primary and secondary prophylaxis of arterial hypertension.

The solution to this problem is provided by the fact that in the method of treatment and prevention of arterial hypertension by introducing a drug into the body based on an activated potentiated form of ultra-low doses of affinity-purified antibodies to the angiotensin II receptor, prepared by repeated serial dilution and external exposure according to the invention, it is additionally simultaneously combined an activated potentiated form of ultra-low doses of affinity-purified antibodies to endothelial NO synth is introduced aze.

An activated potentiated form of antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor II and an activated potentiated form of antibodies to endothelial NO synthase are used.

In addition, the activated potentiated form of antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor and the activated potentiated form of antibodies to endothelial NO synthase are used in the form of an activated potentiated aqueous or aqueous-alcoholic solution of each component obtained by sequential multiple dilution in aqueous or a water-alcohol solvent and an intermediate external mechanical effect - vertical shaking.

In this case, a mixture of various homeopathic dilutions of antibodies to the C-terminal fragment of the AT ₁ receptor of angiotensin II in combination with a mixture of various homeopathic dilutions of antibodies to endothelial NO synthase is prepared in the form of a single drug.

It is possible to use a pharmaceutical composition based on an activated potentiated form of ultra-low doses of affinity-purified antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor and to endothelial NO synthase in combination with known standard drugs used to treat diseases of the cardiovascular system of the following groups: β-blockers, ACE inhibitors, including combined, diuretics, nitrates.

The solution to this problem is also provided by the fact that the drug for the treatment and prevention of arterial hypertension based on the activated potentiated form of ultra-low doses of affinity purified antibodies to the angiotensin II receptor, according to the invention, is made in the form of a pharmaceutical composition and additionally contains an activated potentiated antibody form as an enhancing component to endothelial NO synthase.

In this case, the activated potentiated form of antibodies to the angiotensin II receptor and to endothelial NO synthase is used in the form of an activated potentiated aqueous or hydroalcoholic solution obtained by sequential multiple dilution of a matrix solution of the corresponding antibodies in an aqueous or hydroalcoholic solvent and intermediate external mechanical action - vertical shaking.

In addition, the pharmaceutical composition may be in solid dosage form and contains an effective amount of a neutral carrier saturated with a mixture of an activated potentiated form of antibodies to the angiotensin II receptor and an activated potentiated form of antibodies to endothelial NO synthase, and pharmaceutically acceptable additives that may include lactose, microcrystalline cellulose and magnesium stearate.

In this case, aqueous or aqueous-alcoholic solutions of activated potentiated forms of antibodies to the angiotensin II receptor and to endothelial NO synthase were obtained by repeated serial dilution and intermediate external exposure from matrix solutions of affinity purified antibodies to angiotensin II receptor and to endothelial NO synthase with a concentration of 0 5 ÷ 5.0 mg / ml.

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

In addition, the pharmaceutical composition contains active ingredients: an activated potentiated form of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial NO synthase in a 1: 1 ratio, each component being used in an ultra-low dose as a mixture of three corresponding matrix solutions diluted 100 ¹² , 100 ³⁰ and 100 ²⁰⁰ times, which is equivalent to hundreds of homeopathic dilutions of C 12, C 30, C 200.

It is possible that the claimed pharmaceutical composition based on the activated potentiated form of ultra-low doses of affinity purified antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor and to endothelial NO synthase be used in combination with known standard drugs used to treat diseases of the cardiovascular system of the following groups :

- β-blockers (egilok, atenolol, concor, betalok ZOK);

- ACE inhibitors, including combined (enap, enalapril, kapoten, renitek, prestarium (berlipril, diroton, kapoten, quadropril, monopril, renitek, prestarium, noliprel-forte, enap-N));

- diuretics (furosemide, veroshpiron, hypothiazide, arifon retard, indapamide, hypothiazide, diuver, indap, indapamide);

- cardiac glycosides (digoxin);

- nitrates (dilaside, cardicet, cardicetetordord, mitrolinate, MonoMack, monochinque, nitroglycerin, nitrosorbide, olicardium, pectrol, sydnopharm).

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

The proposed combination of activated potentiated forms of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial synthase nitric oxide (NO synthase) in a pharmaceutical composition (i.e., the forms of antibodies to the C-terminal fragment of AT ₁ receptor for angiotensin II and endothelial NO synthase prepared according to the homeopathic technology of potentiation by repeated serial dilution and intermediate external action - vertical shaking, which are active in the pharmacological model and / or clinical methods of treatment of hypertension) provides an unexpected synergistic therapeutic effect is more pronounced hypotensive effect compared to the effect exerted by the individual components, as confirmed experimentally by the adequate model.

In addition, the claimed drug expands the arsenal of centrally acting drugs intended for treatment, as well as primary and secondary prophylaxis of arterial hypertension without side effects.

The pharmaceutical composition 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 by known technologies - the techniques described, for example, in the book: Immunological methods, ed. G. Fremel, M., "Medicine", 1987, S. 9-33; or, for example, in an article by Laffly E., Sodoyer R. Hum. Antibodies. Monoclonal and recombinant antibodies, 30 years after. - 2005 - Vol.14. - N 1-2. P.33-55.

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

Polyclonal antibodies can be obtained by active immunization of animals. To this end, according to a specially developed scheme, the animals are given a series of injections of the substance required by the invention, an antigen: endothelial NO synthase and the C-terminal fragment of AT ₁ angiotensin II receptor. 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.

Preferred for the preparation of the claimed pharmaceutical composition is the use of polyclonal antibodies to endothelial NO synthase and to the C-terminal fragment of AT ₁ angiotensin II receptor, which are used as a matrix (primary) solution with a concentration of 0.5 ÷ 5.0 mg / ml for subsequent preparation of activated potentiated form.

Preferred for the preparation of each component is the use of a mixture of three water-alcohol dilutions of the primary matrix solution of antibodies diluted 100 ¹² , 100 ³⁰ and 100 ²⁰⁰ times, respectively, which corresponds to hundreds of homeopathic dilutions of C 12, C 30 and C 200. When performing the claimed drug in solid dosage form, a mixture of these components is applied to lactose.

Preferred for the preparation of the claimed drug is the use of polyclonal antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and endothelial NO synthase, which can be obtained by immunization of rabbits as follows.

To obtain polyclonal antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor, the adjuvant and the C-terminal fragment AT ₁ angiotensin II receptor selected, for example, from the following group are used as immunogen (antigen) for rabbits immunization:

GKKFK RYFLQLLKYI PPKAKSHSNL STKMSTLSYR PSDNVSSSTK KPAPCFEVE

LNPF FYVFFGKNFK KYFLQLIKYI PPNVSTHPSL TTKMSSLSYR

PPENIRLPTK

KTAGSFDTE

QLLKYI PPKA

SSLSYRPPENIR

QLIKYI PPNVSTHP

SNL STKMSTLSYR PSDNVSSSTK KPAPCF

It is possible to obtain polyclonal antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor using an angiotensin II human C-terminal fragment AT1 receptor with the cysteine (C) added to the N end as an immunogen (antigen):

CGKKF KRYFL QLLKY IPPKA KSHSN LSTKM STLSY RPSDN VSSST KKRAR CFEVE

Before blood sampling for 1-3 days, 1-3 intravenous injections are performed to increase the level of antibodies. During immunization, small blood samples are taken from rabbits to evaluate the amount of antibody. The maximum level of the immune response to the administration of most soluble antigens is achieved 40-60 days after the first injection. After the end of the first cycle of immunization of rabbits for 30 days, they restore health and re-immunization, including 1-3 intravenous injections. To obtain antisera from immunized rabbits, blood is collected in a 50 ml centrifuge tube. Using a wooden spatula, the formed clots are removed from the walls of the tube and the wand is placed in the clot formed in the center of the tube. Blood is placed in a refrigerator (temperature 4 ° C) at night. The next day, the clot attached to the spatula is removed and the remaining liquid is centrifuged at 13000g for 10 minutes. The supernatant (supernatant) is an antiserum. The resulting antiserum should be yellow. Add to the antiserum 20% (weight concentration) NaN ₃ to a final concentration of 0.02% and store until use in a frozen state at a temperature of -20 ° C. To isolate antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor from antiserum, absorption on the solid phase is carried out in the following sequence:

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

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

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

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

The antibodies are then purified by affinity chromatography by attaching the obtained antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor, which is located on an insoluble matrix, followed by elution with concentrated salt solutions.

Thus obtained buffer solution of polyclonal rabbit antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor purified on antigen with a concentration of 0.5 ÷ 5.0 mg / ml, preferably 2.5 ÷ 3.0 mg / ml, is used in as a matrix (primary) solution for the subsequent preparation of the activated potentiated form.

Polyclonal antibodies to endothelial NO synthase are obtained in a similar manner to the above, using as an immunogen (antigen) for immunization of rabbits an adjuvant and a whole molecule of endothelial NO synthase of the following sequence:

1 MGNLKSVGQE PGPPCGLGLG LGLGLCGKQG PASPAPEPSR ARARATRNAR DHSPAPNSPT

61 LTRPPEGPKF PRVKNWELGS ITYDTLCAQS QQDGPCTPRR CLGSLVLPRK LQTRPSPGPP

121 PAEQLLSQAR DFINQYYSSIKRSGSQAHEE RLQEVEAEVA STGTIHLRES ELVFGAKQAW

181 RNAPRCVGRI QWGKLQVFDA RDCSSAQEMF TYICNHIKYA TNRGNLRSAI TVFPQRAPGR

241 GDFRIWNSQL VRYAGYRQQD GSVRGDPANV EITELCIQHG WTPGNGRFDV LPLLLQAPDE

301 APELFVLPPE LVLEVPLGAP HTGWRGPGL RWYALPAVSN MLLEIGGLEF SAAPFSGWYM

361 STEIGTRNLC DPHRYNILED VAVCMDLDTR TTSSLWKDKA AVEINLAVLH SFQLAKVTIV

421 DHHAATVSFM KHLDNEQKAR GGCPADWAWIVPPIYGSLPP VFHQEMVNYI LSPAFRYQPD

481 PWKGSATKGA GITRKKTFKE VANAVKISAS LMGTLMAKRV KATILYASET GRAQSYAQQL

541 GRLFRKAFDP RVLCMDEYDV VSLEHEALVL WTSTFGNGD PPENGESFAA ALMEMSGPYN

601 SSPRPEQHKS YKIRFNSVSC SDPLVSSWRR KRKESSNTDS AGALGTLRFC VFGLGSRAYP

661 HFCAFARAVD TRLEELGGER LLQLGQGDEL CGQEEAFRGW AKAAFQASCE TFCVGEEAKA

721 AAQDIFSPKR SWKRQRYRLS AQAEGLQLLP GLIHVHRRKM FQATVLSVEN LQSSKSTRAT

781 ILVRLDTAGQ EGLQYQPGDHIGISAPNRPG LVEALLSRVE DPPPPTESVA VEQLEKGSPG

841 GPPPSWVRDP RLPPCTVRQA LTFFLDITSP PSPRLLRLLS TLAEEPSEQQ ELETLSQDPR

901 RYEEWKLVRC PTLLEVLEQF PSVALPAPLL LTQLPLLQPR YYSVSSAPNA HPGEVHLTVA

961 VLAYRTQDGL GPLHYGVCST WLSQLKTGDP VPCFIRGAPS FRLPPDPYVP CILVGPGTGI

1021 APFRGFWQER LHDIESKGLQ PHPMTLVFGC RCSQLDF £ LYR DEVQDAQERG VFGRVLTAFS

1081 REPDSPKTYV QDILRTELAA EVHRVLCLER GHMFVCGDVT MATSVLQTVQ RILATEGDME

1141 LDEAGDVIGV LRDQQRYHED IFGLTLRTQE VTSRIRTQSF SLQERHLRGA VPWAFDPPGP 1201 DTPGP

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

1 MGNLKSVAQE PGPPCGLGLG LGLGLCGKQG PATPAPEPSR APASLLPPAP EHSPPSSPLT

61 QPPEGPKFPR VKNWEVGSIT YDTLSAQAQQ DGPCTPRRCL GSLVFPRKLQ GRPSPGPPAP

121 EQLLSQARDF INQYYSSIKR SGSQAHEQRL QEVEAEVAAT GTYQLRESEL VFGAKQAWRN

181 APRCVGRIQW GKLQVFDARD CRSAQEMFTYICNHIKYATN RGNLRSAITV FPQRCPGRGD

241 FRIWNSQLVR YAGYRQQDGS VRGDPANVEI TELCIQHGWT PGNGRFDVLP LLLQAPDDPP

301 ELFLLPPELV LEVPLEHPTL EWFAALGLRW YALPAVSNML LEIGGLEFPA APFSGWYMST

361 EIGTRNLCDP HRYNILEDVA VCMDLDTRTT SSLWKDKAAV EINVAVLHSY QLAKVTIVDH

421 HAATASFMKH LENEQKARGG CPADWAWIVP PISGSLTPVF HQEMVNYFLS PAFRYQPDPW

481 KGSAAKGTGI TRKKTFKEVA NAVKISASLM GTVMAKRVKA TILYGSETGR AQSYAQQLGR

541 LFRKAFDPRV LCMDEYDVVS LEHETLWLVV TSTFGNGDPP ENGESFAAAL MEMSGPYNSS

601 PRPEQHKSYKIRFNSISCSD PLVSSWRRKR KESSNTDSAG ALGTLRFCVF GLGSRAYPHF

661 CAFARAVDTR LEELGGERLL QLGQGDELCG QEEAFRGWAQ AAFQAACETF CVGEDAKAAA

721 RDIFSPKRSW KRQRYRLSAQ AEGLQLLPGL IHVHRRKMFQ ATIRSVENLQ SSKSTRATIL

781 VRLDTGGQEG LQYQPGDHIG VCPPNRPGLV EALLSRVEDP PAPTEPVAVE QLEKGSPGGP

841 PPGWVRDPRL PPCTLRQALT FFLDITSPPS PQLLRLLSTL AEEPREQQEL EALSQDPRRY

901 EEWKWFRCPT LLEVLEQFPS VALPAPLLLT QLPLLQPRYY SVSSAPSTHP GEIHLTVAVL

961 AYRTQDGLGP LHYGVCSTWL SQLKPGDPVP CFIRGAPSFR LPPDPSLPCI LVGPGTGIAP

1021 FRGFWQERLH DIESKGLQPT PMTLVFGCRC SQLDHLYRDE VQNAQQRGVF GRVLTAFSRE

1081 PDNPKTYVQD ILRTELAAEV HRVLCLERGH MFVCGDVTMA TNVLQTVQRI LATEGDMELD

1141 EAGDVIGVLR DQQRYHEDIF GLTLRTQEVT SRIRTQSFSL QERQLRGAVP WAFEPPGSDT 1201 NSP

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

1192-1195

Pwaf

1189-1192:

Rgavp

1185-1205:

RHLRGAVPWAF DPPGPDTPGP

1194-1205:

AF DPPGPDTPGP

1186-1196:

HLRGAVPWAF D

1186-1205:

HLRGAVPWAF DPPGPDTPGP

The activated potentiated form of each component is prepared by uniformly decreasing the concentration as a result of successive dilution of 1 part of the aforementioned matrix solution in 9 parts (for decimal dilution) or in 99 parts (for hundredth dilution) or in 999 parts (for thousandth dilution) of a neutral solvent with multiple vertical shaking ("dynamization") of each dilution obtained and using separate containers for each subsequent dilution to obtain the desired potency - cr dilution characteristics according to the homeopathic method (see, for example, V. Schwabe "Homeopathic medicines", M., 1967, p.14-29).

For example, to prepare a 12th hundred-fold dilution of C12, one part of the aforementioned matrix solution of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor with a concentration of 3.0 mg / ml is diluted in 99 parts of a neutral aqueous or aqueous-alcoholic solvent (mainly 70% ethyl alcohol) and repeatedly (10 or more times) vertically shake potentiate the obtained 1st hundredth C1 dilution. From the 1st hundredth dilution C1, a second hundredth dilution of C2 is prepared. This operation is repeated 11 times, obtaining the 12th hundredth dilution of C12. Thus, the 12th hundredth dilution of C12 is a solution obtained by diluting successively in different containers 12 times of the 1st part of the initial matrix solution of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor with a concentration of 3.0 mg / ml in 99- te parts of a neutral solvent, i.e. a solution prepared from a matrix solution diluted 100 to ¹² times, which is equivalent to a hundredth homeopathic dilution of C12. Similar operations with an appropriate dilution ratio are carried out to obtain dilutions of C 30 and C 200.

When using as a biologically active liquid component a mixture of various homeopathic, mainly hundred, dilutions of the active substance, each component of the composition (for example, C 12, C 30, C 200) is prepared separately according to the technology described above to the penultimate dilution (respectively, to obtain C 11 , C 29, C 199) and then, in accordance with the composition of the mixture, make one part of each component in one container and mix with the required amount of solvent (respectively, with 97 parts for hundred-percent dilution). In this case, an activated potentiated form of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor is obtained in an ultra-low dose of each component prepared from a matrix solution diluted in 100 ¹² , 100 ³⁰ , 100 ²⁰⁰ , which is equivalent to a mixture of hundred homeopathic dilutions C12, C30 and C200.

It is possible to use the active substance in the form of a mixture of various different homeopathic dilutions, for example decimal and / or hundredths (C 20, C 30, C 100 or C12, C30, C50, etc.), the effectiveness of which is determined experimentally.

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

To obtain the claimed pharmaceutical composition, aqueous or aqueous-alcoholic solutions of the active ingredients are mixed, mainly in a ratio of 1: 1 and used in liquid dosage form. Preferably, the pharmaceutical composition contains an activated potentiated form of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial NO synthase in an ultra-low dose of each component prepared from a matrix solution diluted in 100 ¹² , in 100 ³⁰ , in 100 ²⁰⁰ , which is equivalent a mixture of hundreds of homeopathic dilutions C12, C30 and C200.

The claimed pharmaceutical composition can be used in solid dosage form, which contains an effective amount of granules of a neutral carrier - lactose, saturated by soaking, until saturated with a mixture of aqueous or aqueous-alcoholic solutions, of the activated potentiated form of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and activated potentiated forms of antibodies to endothelial NO synthase, and pharmaceutically acceptable additives, including, mainly, lactose, microcrystalline cellulose matic and magnesium stearate.

To obtain a solid oral form of the claimed medicinal product, a fluidized bed is installed (for example, Huttlin Pilotlab type manufactured by Huttlin GmbH) until the granules of a neutral substance - lactose (milk sugar) are introduced into the fluidized - boiling layer with a particle size of 150 ÷ 300 μm pre-prepared aqueous or aqueous-alcoholic solution of activated potentiated forms of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and endothelial synthase nitric oxide (NO synthase), advantages Specifically, in the ratio of 1 kg of antibody solution to 5 or 10 kg of lactose (1: 5-1: 10) with simultaneous drying in a stream of heated air supplied under the grate at a temperature of no higher than 40 ° C. The estimated amount of 0.17 ÷ 0.34 by weight of the solid oral form of the dried granules saturated with the activated potentiated form of antibodies is loaded into the mixer and mixed with microcrystalline cellulose introduced in an amount of 3-8 wt. hours from the total mass of the load - from the mass of a solid oral form. Then 25 ÷ 45 wt. including the total mass of the load of "unsaturated" pure lactose (to reduce the cost and some simplification and acceleration of the process without reducing the effectiveness of the therapeutic effect), magnesium stearate in an amount of 0.1 ÷ 0.3 wt. including from the total mass of the load and microcrystalline cellulose in an amount of 3 ÷ 8 wt. hours from the total mass of the load. The resulting tablet mass is uniformly mixed and tabletted by direct dry pressing (for example, in a Korsch tablet press - XL 400) with the formation of round tablets weighing 150 ÷ 500 mg. After tabletting, 300 mg tablets are obtained, impregnated with an aqueous-alcoholic solution (3.0-6.0 mg / tablet) of an activated potentiated form of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and NO synthase in an ultra-low dose of each component, prepared from a matrix solution diluted in 100 ¹² , in 100 ³⁰ in 100 ²⁰⁰ , which is equivalent to a mixture of hundreds of homeopathic dilutions C12, C30 and C200.

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

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

Example 1

For an experimental study of the hypotensive effect of the claimed technical solution, a pharmaceutical composition was used containing an aqueous solution of activated potentiated forms (ultra-low doses - SMD) of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial NO synthase, with each component taken in a mixture homeopathic dilutions C12, C30, C200. Studies were conducted on 40 male rats of the SHR line (weight 350 ± 50 g, age 4.5-5 months), caused by hypertension, which were divided into 4 groups of 10 animals.

The following drugs were orally administered to animals once a day for 28 days: group 1 - 2.5 ml / kg of DMD antibodies to the C-terminal fragment of AT ₁ human angiotensin II receptor (a mixture of aqueous dilutions C12, C30, C200) in combination with 2 5 ml / kg of distilled water, group 2 - 2.5 ml / kg of SMD antibodies to endothelial NO synthase (a mixture of aqueous dilutions of C12, C30, C200) in combination with 2.5 ml / kg of distilled water, group 3 - 5 ml / kg pharmaceutical composition based on DMD antibodies to the C-terminal fragment of AT ₁ human angiotensin II receptor and SMD antibodies to endothelial NO c intase (a mixture of aqueous homeopathic dilutions C12, C30, C200 of each component), group 4 - 5 ml / kg of distilled water.

The measurement of systolic blood pressure (SBP) in awake rats was carried out by an indirect method in the caudal artery (using a cuff) once a week 9 hours after the last injection of drugs.

All drugs had a hypotensive effect (p <0.05): by day 28, systolic blood pressure (SBP) decreased compared to the initial level in group 1, which received SMD antibodies to the C-terminal fragment of the human angiotensin II receptor AT ₁ receptor, by 20, 6%; in group 2, receiving SMD antibodies to endothelial NO synthase, by 14.4%; in group 3, which received the combined preparation of SMD antibodies, by 27.6%. In control group 4, the change in SBP relative to baseline was 1.6%.

The obtained results indicate a more pronounced synergistic hypotensive effect of the claimed pharmaceutical composition containing activated potentiated forms of ultra-low doses of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial NO synthase in SHR rats compared to animals treated with individual components ( SMD antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor or SMD antibodies to endothelial NO synthase).

Example 2

For an experimental study of the hypotensive effect of the claimed technical solution, a pharmaceutical composition was used containing an aqueous solution of activated potentiated forms (ultra low doses - SMD) of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial NO synthase, with each component taken in a mixture homeopathic dilutions C12, C30, C200. The studies were conducted on 50 ISIAH male rats (weight 300 g, age 4 months) with hereditary stress-sensitive arterial hypertension, which were divided into 5 groups of 10 animals.

Animals orally 1 time per day for 28 days received the following drugs: group 1 - 2.5 ml / kg of SMD antibodies to the C-terminal fragment of AT ₁ human angiotensin II receptor (a mixture of dilutions C12, C30, C200) in combination with 2, 5 ml / kg of distilled water, group 2 - 2.5 ml / kg of SMD antibodies to endothelial NO synthase (mixture of dilutions C12, C30, C200) in combination with 2.5 ml / kg of distilled water, group 3-5 ml / kg of a pharmaceutical composition based on DMD antibodies to the C-terminal fragment of AT ₁ human angiotensin II receptor and SMD antibodies to endothelial NO synthase (mixture homeopathic aqueous dilutions C12, C30, C200 of each component), group 4 - 5 ml / kg (dose 10 mg / kg) comparison drug losartan, group 5 - 5 ml / kg of distilled water.

Twice a week 2–6 hours after the administration of SMD antibodies and losartan, systolic blood pressure (SAD) was measured indirectly in the caudal artery (using a cuff). Table 1 shows the dynamics of changes in systolic blood pressure in ISIAH rats, as measured by the indirect method.

Table 1 Indicator GARDEN outcome., In
mmHg. GARDEN after 28 days of drug administration, in mmHg Δ compared with the initial level, in mm Hg % of baseline SMD of antibodies to the C-terminal fragment of AT ₁ human angiotensin II receptor 176 150 -26 -14.7% SMD antibodies to endothelial NO synthase 175 164.5 -10.5 -6% Combined preparation based on SMD antibodies to the C-terminal fragment of the ATI angiotensin II receptor and to endothelial NO synthase 179.5 140 -39.5 -22% Losartan 173.5 140.5 -33 -19% Control (distilled water) 181 178 -3 -1.6%

The results indicate a more pronounced synergistic hypotensive effect of the claimed pharmaceutical composition containing activated potentiated forms of ultralow doses of antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor and to endothelial NO synthase in ISIAH rats, compared with animals treated with individual components ( SMD antibodies to the C-terminal fragment of AT ₁ angiotensin II receptor or SMD antibodies to endothelial NO synthase) combinations. Moreover, the claimed pharmaceutical composition containing activated potentiated forms of ultra-low doses of antibodies to the C-terminal fragment of AT1 of the angiotensin II receptor and to endothelial NO synthase is at least as effective as the well-known and widely used antihypertensive drug losartan.

Claims (11)

1. A method for the treatment and prevention of arterial hypertension by introducing into the body a drug based on an activated potentiated form of affinity-purified antibodies to the angiotensin II receptor, prepared by repeated serial dilution and external exposure, characterized in that the activated potentiated form of affinity-purified is additionally simultaneously combined. antibodies to endothelial NO synthase. 2. The method according to claim 1, characterized in that they use an activated potentiated form of antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor II and an activated potentiated form of antibodies to endothelial NO synthase. 3. The method according to claim 1 or 2, characterized in that the activated potentiated form of antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor II and the activated potentiated form of antibodies to endothelial NO synthase are used in the form of an activated potentiated aqueous or alcoholic solution each component obtained in the process of serial multiple dilutions in an aqueous or hydroalcoholic solvent and an intermediate external mechanical action - shaking of each dilution. 4. The method according to claim 1 or 2, characterized in that they use a pharmaceutical composition prepared in the form of a single drug - one dosage form, which includes a mixture of various homeopathic dilutions of antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor II in combination with a mixture of different homeopathic dilutions of antibodies to endothelial NO synthase. 5. The method according to claim 1 or 2, characterized in that they use a pharmaceutical composition based on an activated potentiated form of affinity-purified antibodies to the C-terminal fragment of the AT ₁ angiotensin II receptor and to endothelial NO synthase in combination with known standard drugs, used for the treatment of diseases of the cardiovascular system of the following groups: ACE inhibitors, including combined, diuretics, β-blockers, nitrates, cardiac glycosides, calcium antagonists, lipid-lowering drugs, antiplatelet agents, antihypoxants, anticoagulants. 6. The drug for the treatment of arterial hypertension according to claim 1, based on an activated potentiated form of affinity purified antibodies to the angiotensin II receptor, characterized in that it is made in the form of a pharmaceutical composition and further comprises an activated potentiated form of antibodies to endothelial NO synthase as a reinforcing component . 7. The drug according to claim 6, characterized in that the activated potentiated form of antibodies to the angiotensin II receptor and to endothelial NO synthase is used in the form of an activated potentiated aqueous or aqueous-alcoholic solution obtained by sequential multiple dilution of the matrix solution of the corresponding antibodies in aqueous or hydroalcoholic solvent and intermediate external mechanical action - shaking each dilution. 8. The drug according to claim 6 or claim 7, characterized in that the pharmaceutical composition is made in solid dosage form and contains an effective amount of granules of a neutral carrier with an activated potentiated form of antibodies to the angiotensin II receptor and an activated potentiated form of antibodies to endothelial NO synthase and pharmaceutically acceptable additives. 9. The drug according to claim 6 or 7, characterized in that aqueous or aqueous-alcoholic solutions of activated potentiated forms of antibodies to the angiotensin II receptor and to endothelial NO synthase are obtained by repeated serial dilution and intermediate external exposure from matrix solutions of affinity purified antibodies to the angiotensin II receptor and to endothelial NO synthase with a concentration of 0.5-5.0 mg / ml. 10. The drug according to claim 6 or 7, characterized in that each of the components of the affinity purified antibodies is used as a mixture of various, mainly hundred, homeopathic dilutions. 11. The drug of claim 8, characterized in that the pharmaceutically acceptable additives include lactose, microcrystalline cellulose and magnesium stearate.