SE1350184A1

SE1350184A1 - PROCEDURE TO INCREASE THE EFFECT OF AN ACTIVATED, POTENTIZED FORM OF AN ANTIBODY

Info

Publication number: SE1350184A1
Application number: SE1350184A
Authority: SE
Inventors: Oleg Iliich Epshtein
Original assignee: Oleg Iliich Epshtein
Priority date: 2010-07-15
Filing date: 2011-07-15
Publication date: 2013-04-12

Abstract

ABSTRACT The preset invention provides a method of increasing the effect of an activated-potentiatedform of an antibody to an endogenous biological molecule by combining said endogenousbiological molecule with an activated-potentiated form of an antibody to endothelial NO-synthase. The present invention also provides a pharmaceutical composition comprising a) anactivated-potentiated form of an antibody to an endogenous biological molecule, and b) anactivated-potentiated form of an antibody to NO synthase. 90

Description

A METHOD OF INCREASING THE EFFECT OF AN ACTIVATED-POTENTIATED FORM OF AN ANTIBODY FIELDThe present invention relates to a method of increasing the effect of an activated-potentiatedform of an antibody and a pharmaceutical formulation comprising an activated-potentiated form ofan antibody to an endogenous biological molecule and an activated-potentiated form of an antibodyto endothelial NO-synthase.

BACKGROUND Nitric oxide (NO) is a gaseous molecule that has been shown to acts in thesignaling of different biological processes. Endothelium-derived NO is a key molecule inregulation of vascular tone and its association with vascular disease has long beenrecognized. NO inhibits many processes known to be involved in the formation ofatherosclerotic plaque, including monocyte adhesion, platelet aggregation and vascularsmooth muscle cell proliferation. Another important role of endothelial NO is theprotection of the vascular wall from the oxidative stress induced by its own metabolicEndothelial dysfunction occurs at very early stages of atherosclerosis. lt is therefore possible that products and by the oxidation products of lipids and lipoproteins. deficiency in local NO availability could be a final common pathway that acceleratesatherogenesis in humans. ln addition to its role in the vascular endothelium, NOavailability has been shown to modulate metabolism of lipoproteins. Negativecorrelation has been reported between plasma concentrations of NO metabolic productsand plasma total and Low Density Lipoprotein [LDL] cholesterol levels while HighDensity Lipoprotein [HDL] improves vascular function in hypercholesterolaemic subjects.The loss of NO has considerable effect on the development of the disease. Diabetesmellitus is associated with increased rates of morbidity and mortality caused primarily bythe accelerated development of atherosclerotic disease. l\/loreover, reports show that diabetics have impaired lung functions. lt has been proposed that insulin resistance leads to airvvay inflammation. Habib et al., Nitric Oxide Measurement From Blood To Lungs, Is There A Link? Pak J Physiol 2007; 3(1).

Nitric oxide is synthesized by the endothelium from L-arginine by nitric oxidesynthase (NO synthase). NO synthase occurs in different isoforms, including aconstitutive form (cNOS) and an inducible form (iNOS). The constitutive form is presentin normal endothelial cells, neurons and some other tissues.

The therapeutic effect of an extremely diluted form (or ultra-low form) ofantibodies potentized by homeopathic technology (activated potentiated form) has beenOleg l. U.S. Patent No. 7,700,096 discloses ahomeopathically potentized form of antibodies to endothelial NO-synthase. The homeopathically potentized form of antibodies to endothelial NO-synthase is marketed in discovered by Dr. Epshtein. the Russian Federation and other countries under the name lmpaza®.There is a continuing need for a method of increasing the effect of an activated- potentiated form of an antibody.

SUMMARY ln accordance with one aspect, the present invention provides a method ofincreasing the effect of an activated-potentiated form of an antibody to an endogenousbiological molecule, said method comprising combining said endogenous biologicalmolecule with an activated-potentiated form of an antibody to endothelial NO-synthase.Preferably, the method aspect of the invention includes administering said combinationto a patient in need of treatment with said activated-potentiated form of an antibody. ln one variant, said activated-potentiated form of an antibody to an endogenousbiological molecule is an antibody to S-1OO protein. ln another variant, said activated-potentiated form of an antibody to an endogenous biological molecule is an antibody toprostate specific antigen. ln another variant, said activated-potentiated form an antibodyto an endogenous biological molecule is an antibody to insulin receptor. ln anothervariant, said activated-potentiated form an antibody to an endogenous biologicalmolecule is an antibody to antigiotensin receptor ll. ln accordance with another aspect, the invention provides a pharmaceuticalcomposition comprising a) an activated-potentiated form of an antibody to anendogenous biological molecule, and b) an activated-potentiated form of an antibody toNO synthase. Preferably, the pharmaceutical composition pharmaceutically acceptablesolid carrier. Preferably, the activated-potentiated form of an antibody to endothelial NOsynthase contains a mixture of C12, C30, and C200 homeopathic dilutions impregnatedonto the solid carrier. The activated-potentiated form of an antibody to an endogenousbiological molecule could be a monoclonal, monoclonal, or natural antibody. Preferably,the antibody to human insulin receptor is a polyclonal antibody. lt is contemplated that the pharmaceutical composition comprises an activated-potentiated form of an antibody to an endogenous biological molecule prepared bysuccessive centesimal dilutions coupled with shaking of every dilution. lt is alsocontemplated that the antibody to endothelial NO-synthase is monoclonal, polyclonal ornatural antibody. lt is particularly preferred that the antibody to endothelial NO-synthaseis a polyclonal antibody. lt is contemplated that the activated-potentiated form of anantibody to endothelial NO-synthase is prepared by successive centesimal dilutions coupled with shaking of every dilution.

DESCRIPTION OF THE FIGURESFigure 1 - lllustrates the effect of tested preparations on blood plasma glucose level ofrats with streptozotocin-induced diabetes mellitus Figure 2 - lllustrates the effect of tested preparations on day 14 of injection on indicatorsof area under concentration-time curve (AUC) in the glucose tolerance test in rats withstreptozotocin-induced diabetes mellitus.

Figure 3 - lllustrates the effect of tested preparations on blood plasma glucose level of rats with spontaneous non-insulin-dependent diabetes mellitus.

Figure 4 - lllustrates the effect of tested preparations on day 28 of injection on indicatorsof area under concentration-time curve (AUC) in glucose tolerance test in rats withspontaneous non-insulin-dependent diabetes mellitus. 5 DETAILED DESCRIPTION The invention is defined with reference to the appended claims. With respect tothe claims, the glossary that follows provides the relevant definitions.

The term “antibody” as used herein shall mean an immunoglobulin thatspecifically binds to, and is thereby defined as complementary with, a particular spatialand polar organization of another molecule. Antibodies as recited in the claims mayinclude a complete immunoglobulin or fragment thereof, may be natural, polyclonal ormonoclonal, and may include various classes and isotypes, such as lgA, lgD, lgE, lgG1,lgG2a, lgG2b and lgG3, lgl\/l, etc. Fragments thereof may include Fab, Fv and F(ab')2,Fab', and the like. The singular “antibody” includes plural “antibodies.” The term “activated-potentiated form” or “potentiated form” respectively, withrespect to antibodies recited herein is used to denote a product of homeopathicpotentization of any initial solution of antibodies. “Homeopathic potentization” denotesthe use of methods of homeopathy to impart homeopathic potency to an initial solutionof relevant substance. Although not so limited, 'homeopathic potentization” may involve,for example, repeated consecutive dilutions combined with external treatment,particularly vertical (mechanical) shaking. ln other words, an initial solution of antibodyis subjected to consecutive repeated dilution and multiple vertical shaking of eachobtained solution in accordance with homeopathic technology. The preferredconcentration of the initial solution of antibody in the solvent, preferably water or a water-ethyl alcohol mixture, ranges from about 0.5 to about 5.0 mg/ml. The preferredprocedure for preparing each component, i.e. antibody solution, is the use of the mixtureof three aqueous or aqueous-alcohol dilutions of the primary matrix solution (mothertineture) ef antiheeies diiuted 10012, 10000 anei 100200 times, respeetiveiy, whieh isequivalent to centesimal homeopathic dilutions (C12, C30, and C200) or the use of themixture of three aqueous or aqueous-alcohol dilutions of the primary matrix solution ofantibodies diiuted 10012, 10000 anei 10000 times, respeetiveiy, whieh is equivalent te centesimal homeopathic dilutions (C12, C30 and C50). Examples of homeopathic potentization are described in U.S. Patent. Nos. 7,572,441 and 7,582,294, which areincorporated herein by reference in their entirety and for the purpose stated. While theterm “activated-potentiated form” is used in the claims, the term “ultra-low doses” is usedin the examples. The term “ultra-low doses” became a term of art in the field of artcreated by study and use of homeopathically diluted and potentized form of substance.The term “ultra-low dose” or “ultra-low doses” is meant as fully supportive and primarilysynonymous with the term 'activated-potentiated” form used in the claims. ln other words, an antibody is in the “activated-potentiated” or “potentiated” formwhen three factors are present. First, the “activated-potentiated” form of the antibody isa product of a preparation process well accepted in the homeopathic art. Second, the“activated-potentiated” form of antibody must have biological activity determined bymethods well accepted in modern pharmacology. And third, the biological activityexhibited by the “activated potentiated” form of the antibody cannot be explained by thepresence of the molecular form of the antibody in the final product of the homeopathicprocess.

For example, the activated potentiated form of antibodies may be prepared bysubjecting an initial, isolated antibody in a molecular form to consecutive multiple dilutionscoupled with an external impact, such as mechanical shaking. The external treatment in thecourse of concentration reduction may also be accomplished, for example, by exposure toultrasonic, electromagnetic, or other physical factors. V. Schwabe "Homeopathic medicines",l\/l., 1967, U.S. Patents Nos. 7,229,648 and 4,311,897, which are incorporated by reference intheir entirety and for the purpose stated, describe such processes that are well-acceptedmethods of homeopathic potentiation in the homeopathic art. This procedure gives rise to auniform decrease in molecular concentration of the initial molecular form of the antibody. Thisprocedure is repeated until the desired homeopathic potency is obtained. For the individualantibody, the required homeopathic potency can be determined by subjecting theintermediate dilutions to biological testing in the desired pharmacological model. Althoughnot so limited, 'homeopathic potentization” may involve, for example, repeatedconsecutive dilutions combined with external treatment, particularly vertical (mechanical) shaking. ln other words, an initial solution of antibody is subjected to consecutive repeated dilution and multiple vertical shaking of each obtained solution in accordancewith homeopathic technology. The preferred concentration of the initial solution ofantibody in the solvent, preferably, water or a water-ethyl alcohol mixture, ranges fromabout 0.5 to about 5.0 mg/ml. The preferred procedure for preparing each component,i.e. antibody solution, is the use of the mixture of three aqueous or aqueous-alcoholdilutions of the primary matrix solution (mother tincture) of antibodies diluted 10012, 10030and 100200 times, respectively, which is equivalent to centesimal homeopathic dilutionsC12, C30 and C200 or the mixture of three aqueous or aqueous-alcohol dilutions of theprimary matrix solution (mother tincture) of antibodies diluted 10012, 10030 and 10050times, respectively, which is equivalent to centesimal homeopathic dilutions C12, C30and C50. Examples of how to obtain the desired potency are also provided, for example, inU.S. Patent Nos. 7,229,648 and 4,311,897, which are incorporated by reference for thepurpose stated. The procedure applicable to the “activated-potentiated” form of theantibodies described herein is described in more detail below.

There has been a considerable amount of controversy regarding homeopathictreatment of human subjects. While the present invention relies on acceptedhomeopathic processes to obtain the “activated-potentiated” form of antibodies, it doesnot rely solely on homeopathy in human subjects for evidence of activity. lt has beensurprisingly discovered by the inventor of the present application and amplydemonstrated in the accepted pharmacological models that the solvent ultimatelyobtained from consecutive multiple dilution of a starting molecular form of an antibodyhas definitive activity unrelated to the presence of the traces of the molecular form of theantibody in the target dilution. The “activated-potentiated” form of the antibody providedherein are tested for biological activity in well accepted pharmacological models ofactivity, either in appropriate in vitro experiments, or in vivo in suitable animal models.The experiments provided further below provide evidence of biological activity in suchmodels. Human clinical studies also provide evidence that the activity observed in theanimal model is well translated to human therapy. Human studies have also provided evidence of availability of the “activated potentiated” forms described herein to treat specified human diseases or disorders well accepted as pathological conditions in themedical science.

Also, the claimed “activated-potentiated” form of antibody encompasses onlysolutions or solid preparations the biological activity of which cannot be explained by thepresence of the molecular form of the antibody remaining from the initial, startingsolution. ln other words, while it is contemplated that the “activated-potentiated” form ofthe antibody may contain traces of the initial molecular form of the antibody, one skilledin the art could not attribute the observed biological activity in the acceptedpharmacological models to the remaining molecular form of the antibody with anydegree of plausibility due to the extremely low concentrations of the molecular form ofthe antibody remaining after the consecutive dilutions. While the invention is not limitedby any specific theory, the biological activity of the “activated-potentiated” form of theantibodies of the present invention is not attributable to the initial molecular form of theantibody. Preferred is the “activated-potentiated” form of antibody in liquid or solid formin which the concentration of the molecular form of the antibody is below the limit ofdetection of the accepted analytical techniques, such as capillary electrophoresis andHigh Performance Liquid Chromatography. Particularly preferred is the “activated-potentiated” form of antibody in liquid or solid form in which the concentration of themolecular form of the antibody is below the Avogadro number. ln the pharmacology ofmolecular forms of therapeutic substances, it is common practice to create a dose-response curve in which the level of pharmacological response is plotted against theTheminimal level of the drug which produces any detectable response is known as a concentration of the active drug administered to the subject or tested in vitro.threshold dose. lt is specifically contemplated and preferred that the “activated-potentiated” form of the antibodies contains molecular antibody, if any, at aconcentration below the threshold dose for the molecular form of the antibody in the given biological model.

The combination pharmaceutical composition in accordance with this aspect ofthe invention may be in the liquid form or in solid form. Each of the activated potentiated forms of the antibodies included in the pharmaceutical composition is prepared from aninitial molecular form of the antibody via a process accepted in homeopathic art. Thestarting antibodies may be monoclonal, or polyclonal antibodies prepared in accordancewith known processes, for example, as described in lmmunotechniques, G. Frimel, l\/l.,“lVleditsyna”, 1987, p. 9-33; “Hum. Antibodies. l\/lonoclonal and recombinant antibodies,30 years after” by Laffly E., Sodoyer R. - 2005 - Vol. 14. - N 1-2. P.33-55, bothincorporated herein by reference. l\/lonoclonal antibodies may be obtained, e.g., by means of hybridoma technology.The initial stage of the process includes immunization based on the principles alreadydeveloped in the course of polyclonal antisera preparation. Further stages of workinvolve the production of hybrid cells generating clones of antibodies with identicalspecificity. Their separate isolation is performed using the same methods as in the caseof polyclonal antisera preparation.

Polyclonal antibodies may be obtained via active immunization of animals. Forthis purpose, for example, suitable animals (e.g. rabbits) receive a series of injections ofthe appropriate antigen, for example, NO synthase. The animals' immune systemgenerates corresponding antibodies, which are collected from the animals in a knownmanner. This procedure enables preparation of a monospecific antibody-rich serum. lf desired, the serum containing antibodies may be purified, for example by usingaffine chromatography, fractionation by salt precipitation, or ion-exchangechromatography. The resulting purified, antibody-enriched serum may be used as astarting material for the preparation of the activated-potentiated form of the antibodies.The preferred concentration of the resulting initial solution of antibody in the solvent,preferably water or a water-ethyl alcohol mixture, ranges from about 0.5 to about 5.0mg/ml.

The preferred procedure for preparing each component of the combination drugaccording to the present invention is the use of the mixture of three aqueous-alcoholdilutions of the primary matrix solution of antibodies diluted 10012, 10030 and 10050 times,respectively, which is equivalent to centesimal homeopathic dilutions C12, C30, and C50 or diluted 10012, 10030 and 100200 times, respectively, which is equivalent to centesimal homeopathic dilutions C12, C30 and C200. To prepare a solid dosage form, a solidcarrier is treated with the desired dilution obtained via the homeopathic process. Toobtain a solid unit dosage form of the combination of the invention, the carrier mass isimpregnated with each of the dilutions. Both orders of impregnation are suitable toprepare the desired combination dosage form. ln a preferred embodiment, the starting material for the preparation of theactivated potentiated form that comprise the combination of the invention is polyclonal,animal-raised antibody to the corresponding antigen, namely, NO synthase andendogenous biological molecule. To obtain the activated-potentiated form of polyclonalantibodies to NO synthase, the desired antigen may be injected as immunogen into alaboratory animal, preferably, rabbits. Polyclonal antibodies to NO synthase may be obtained using the whole molecule of bovine NO synthase of the following sequence: SEQ ID NO: 1Met Gly Asn Leu Lys Ser Val Gly Gln Glu Pro Gly Pro Pro Cys1 5 10 15Gly Leu Gly Leu Gly Leu Gly Leu Gly Leu Cys Gly Lys Gln Gly16 20 25 30Pro Ala Ser Pro Ala Pro Glu Pro Ser Arg Ala Pro Ala Pro Ala31 35 40 45Thr Pro His Ala Pro Asp His Ser Pro Ala Pro Asn Ser Pro Thr46 50 55 60Leu Thr Arg Pro Pro Glu Gly Pro Lys Phe Pro Arg Val Lys Asn61 65 70 75Trp Glu Leu GLys er Ile Thr Tyr Asp Thr Leu Cys Ala Gln Ser76 80 85 90Gln Gln Asp Gly Pro Cys Thr Pro Arg Cys Cys Leu GLys er Leu91 95 100 105 Val Leu Pro Arg Lys Leu Gln Thr Arg Pro Ser Pro Gly Pro Pro 106 110 115 120Pro Ala Glu Gln Leu Leu Ser Gln Ala Arg Asp Phe Ile Asn Gln121 125 130 135 Tyr Tyr Ser Ser Ile Lys Arg Ser GLys er Gln Ala His Glu Glu136 140 145 150Arg Leu Gln Glu Val Glu Ala Glu Val Ala Ser Thr Gly Thr Tyr151 155 160 165His Leu Arg Glu Ser Glu Leu Val Phe Gly Ala Lys Gln Ala Trp166 170 175 180 Arg181Gln196Thr211Leu226Gly241Tyr256Glu271Gly286Ala301Pro316Arg331Gly346Ser361Asn376Thr391Leu406Asp421Glu436Val451Met466Pro481Lys496Leu511 AsnValTyrArgAspArgIleArgProLeuTrpGlyThrIleThrAlaHisGlnProValTrpThr Met AlaPheIleSerPheGlnThrPheGluGluTyrLeuGluLeuSerValHisLysProAsnLysPhe Gly ProAspCysAlaArgGlnGluAspLeuHisAlaGluIleGluSerLeuAlaAlaIleTyrGLyLys Thr Arg185Ala200Asn215Ile230Ile245Asp260Leu275Val290Phe305Pro320Leu335Phe350Gly365Asp380Leu395His410Ala425Arg440Ser455Ile470Ser485Glu500Leu515 CysArgHisThr Trp ValAspIleVal Asn GLys er CysLeuValThrProSerThrValTrpSer Thr IleProLeuLeuAla Ala AlaLysPhe Val Gly Gly GLys er Leu Ala Val Met Ser Thr Ala Ala GlyCysLysPheSerValGlnLeuProGluValAlaAsnValAspGlnSerCysLeuProLysAsn Lys ArgSerTyrProGlnArgHisLeuProTrpSerProLeuCysLysLeuPheProThrAlaGlyAla Arg Ile190Ser205Ala220Gln235Leu250Gly265Gly280Leu295Glu310Phe325Asn340Phe355Cys370Met385Ala400Ala415Met430Ala445Pro460Phe475Ala490Val505Val510 GlnAlaThrArgValAspTrpGlnLeuAlaMet Ser AlaLys Lys ValArgGlyLys Lys TrpGlnAsn Ala ProThrAlaValAlaLeuGlyProLeuValValHisTrpPheTyrIleIle Ala GlyGluArgProTyrAlaProProLeuLeuLeuTrpHisAspGluThrLeuAlaHisGlnThrSer Thr LysMetGlyGlyAlaAsnGlyAspGluGlyGlu Tyr ThrIleIleAspTrpGlnProArgAla Ile Leu195Phe210Asn225Arg240Gly255Val270Asn285Glu300Val315Leu330Ile345Met360Tyr375Arg390Asn405Val420Asn435Ile450Glu465Asp480Lys495Ser510Leu525 Tyr526Gly541Asp556Val571Glu586Ser601Asn616Lys631Thr646His661Leu676Cys691Gln706Ala721Arg736Gly751Leu766Ile781Gln796Leu811Thr826Gly841Thr856 AlaArgGluValSerSerSerArgLeuPheGlyGlyAlaAlaTyrLeuSerLeuProValGluPro Leu SerLeuTyrThrPheProVal Lys CysGlyGlnSerGlnArgIleValValGlyGluSerPro Arg GluPheAspSerAlaArgSerGluPheAlaGluGluCysAspLeuHis Glu AspAlaValPro Gln Thr530Arg545Val560Thr575Ala590Pro605Cys620Ser635Cys650Phe665Arg680Glu695Glu710Ile725Ser740Val755Asn770Leu785His800Leu815Ala830Ser845Ala860 GlyLysValPheAlaGluSerSerValAlaLeuAlaThrPheThrHisLeuAspIleLeuValTrp Leu ArgAlaSerGlyLeuGlnAspAsnPheArgLeuPhePheSerGlnArgGlnThrGlySerGluVal Thr AlaPheLeuAsnMetHisProThrGlyAlaGlnArgCysProAlaArgSerAlaIleArg Gln Phe 11 GlnAspGluGlyGluLysLeuAspLeuValLeuGlyValLysGluLysSerGlyCysValLeuAsp Phe Ser535Pro550His565Asp580Met595Ser610Val625Ser640GLy655Asp670Gly685Trp700Gly715Arg730Gly745Met760Lys775Gln790Pro805Glu820Glu835Pro850Leu865 TyrArgGluProSerTyrSerAlaSerThrGlnAlaGluSerLeuPheSerGluProAsp Lys Asp AlaValAlaProGlyLysSer Gly ArgGlyLysGluTrpGlnGlnThrGlyAsn Pro GlnLeuLeuGluProIleTrpAlaAlaLeuAspAlaAlaLysLeuAlaArgLeuArg Pro GLys er Leu Ile Pro Thr GlnCysValAsnTyrArgArgLeuTyrGluGluAla Lys LeuThrAlaGlnProProProPro Ser Leu540Met555Leu570Gly585Asn600Phe615Arg630Gly645Pro660Glu675Leu690Phe705Ala720Gln735Pro750Val765Thr780Tyr795Gly810Pro825Gly840Cys855Pro870 Pro Ser871Pro886Arg901Val916Leu931Ser946Val961Gly976Val991Asp1006Ala Pro1021Ser Lys1036Arg Cys1051Ala Gln1066Arg Glu1081Thr Glu1096Gly His1111Leu Gln1126Leu Asp1141Arg Tyr1156Val Thr1171His Leu1186Asp Thr1201 SerTyrLeuThrAlaLeuValPro Pro ProGluGluGluGlnProAlaCysCysTyrPheGlySerGluProLeuMetThrGluHisSerArg Pro Leu875Gln890Trp905Phe920Pro935Ala950Arg965Thr980Ile995Val Pro 1010Arg Gly 1025Leu Gln 1040Gln Leu 1055Arg Gly 1070Asp Ser 1085Ala Ala 1100Phe Val 1115Val Gln 1130Ala Gly 1145Glu Asp 1160Arg Ile 1175Gly Ala 1190Gly Pro1205 GlnGluGlnLeuAsnTyrSer Phe LeuGluLysProLeuHisThrTrpArgCysPheProAspValProGluCysArgAspIleArg Val LeuTrpSerLeuProGln Leu IleTrpAlaHisPheLysValGlyIleValPheThr Pro LeuGluPheValGlnGlyAspSerAlaLeuGlnProLeuGlyThrHisAspLeuIleGlyGln Trp 12 Ser Thr880Leu895Cys910Leu925Arg940Val955Leu970Leu985Pro Ser1000Val Gly1015 Glu Arg Leu1030Met Thr1045Tyr Arg1060Arg Val1075Tyr Val1090Arg Val1105Val Thr1120Ala Thr1135Gly Val1150Leu Thr1165Ser Phe1180Ala Phe1195 LeuThr SerArg ProAla ProPro TyrGlu HisGly GlyGln LysPhe Pro LeuAspLeuGlnLeuMetGluLeuLeu Ser LeuGlnThrAlaTyrLeuPro Thr GlyHisValGluThrAspCysAlaGlyArgArgLeu Pro AlaAspLeuProSerThrLeuGlyLeuThrAspPheValAlaIleLeuThrAspAspThrGln Pro Glu Glu885Arg900Glu915Leu930Ser945Ala960Tyr975Pro990Pro Pro1005Gly Ile1020Ile Glu1035Gly Cys1050Gln Asp1065Phe Ser1080Leu Arg1095Glu Arg1110Ser Val1125Met Glu1140Gln Gln1155Gln Glu1170Glu Arg1185Gly Pro1200 ProLeuLeuValValHis Asp Polyclonal antibodies to NO synthase may be obtained using the whole molecule of human NO synthase of the following sequence: SEQ ID NO:2 Met1Gly16Pro31Leu46Gln61Val76Asp91Pro106Glu121Ser136 Gln Gly AsnLeu GlyThr Ala Pro Pro Pro Pro GLys er Pro Gly Arg Lys Gln Leu Ser Ile Glu Val Leu Leu Pro Ala Glu Ile Cys Leu Leu Lys Glu SerLeuAla ProPro GluGly ProThr Tyr80Thr ProGln Gly110Ser Gln125Arg Ser140 Ala Glu Val Gly Glu His Lys Asp Arg Arg Ala Ala Leu Pro Ser Phe Thr Arg Pro Arg GLys er Val Ala 13 Gln Gly Ser Pro Pro Leu Cys Ser Asp Gln Ala Glu Leu Arg Pro Arg Ser Leu100Pro115Phe130Ala145 Thr Pro Cys Ala Ser Val Ala Gly Gly Pro Ser Lys Gln GLys er Gly Pro Ile Asn His Glu Gly Thr Pro Lys Ala Pro Asn Ala Leu Pro Gln Gln Tyr Pro Gln Ser Leu Trp Gln Val Ala Tyr Arg Gln Cys Gly Leu Thr Glu Gln 90Phe105Pro120Tyr135Leu150 Leu 151Arg166Ala181Phe196Ile211Ser226Phe241Gln256Thr271Phe286Glu301Glu316Tyr 331 Glu Pro Asp Cys Ala Arg Gln Glu Asp Leu His Ala Ser Arg Ala Asn Ile Ile Asp Leu Val Phe Pro Leu Glu Cys Arg His Thr Trp GLy Cys Leu Leu Thr Pro 155Leu170Val185Asp200Ile215Val230Asn245Ser260Ile275Pro290Leu305Leu320Ala 335 Val Gly Cys Lys Phe Ser Val Gln Leu Pro Glu Val Phe Arg Arg Tyr Pro Gln Arg His Leu Pro Trp Ser Gly Ile Ser Ala Gln Leu Gly Gly Leu Glu Phe Asn 14 Ala Gln Ala Thr Arg Val Asp Trp Gln Leu Ala Met 160Lys175Trp190Gln205Asn220Cys235Arg250Pro265Thr280Ala295Val310Ala325Leu 340 Gln Gly Glu Arg Pro Tyr Ala Pro Pro Leu Leu Leu Ala Lys Met Gly Gly Ala Asn Gly Asp Glu Gly Glu Trp Leu Phe Asn Arg Gly Val Asn Glu Val Leu Ile Arg Gln Thr Leu Gly Tyr Glu Gly Pro Pro Arg Gly 165Asn180Val195Tyr210Arg225Asp240Arg255Ile270Arg285Pro300Leu3l5Trp330Gly 345 Leu346Glu361Leu376Ser391Val406His421Lys436Pro451Asn466Lys481Phe496Gly511 Ser Glu Ile Glu Ser Leu Ala Ala Ile Tyr Gly Lys Thr Glu Phe Gly Asp Leu His Ala Arg Ser Phe Ser Glu Val Thr Pro Ala 350Thr Arg365Val Ala380Trp Lys395Ser Tyr410Thr Ala425Gly 440GLys er455Leu Ser 470Ala Ala485Val Ala500Met Ala515 Gly Arg Ala Asn Val Asp Gln Ser Cys Leu Pro Lys Asn Lys Ala Pro Leu Cys Lys Leu Phe Pro Thr Ala Gly Ala Arg Gln Phe Cys Met Ala Ala Met Ala Pro Phe Thr Val Val Ser Ser Asp Asp Ala Lys Lys Asp Val Arg Gly Lys Lys Tyr Gly355Pro370Leu385Val400Val415His430Trp445Phe460Tyr475Ile490Ile505Ala510 Ala Trp His Asp Glu Thr Leu Ala His Gln Thr Ser Thr Gln Tyr Arg Thr Ile Ile Glu Trp Gln Pro Arg Ala Ile Gln Met Tyr Arg Asn Val Asn Ile Glu Asp Lys Ser Leu Leu Ser Asn Thr Val Asp Glu Val Met Pro Lys Leu Tyr Gly Thr360Ile375Thr390Ala405His420Gln435Pro450Val465Trp480Thr495Met510Gly525 Arg 526Leu541Tyr556Thr571Phe586Pro601Ile616Lys631Arg646Cys661Gly676Gln691Ala 706 Phe Asp Ser Ala Arg Ser Glu Phe Ala Glu Glu Cys Arg Val Thr Ala Pro Cys Ser Cys Phe Arg Glu Glu Lys Val Phe Ala Glu Ser Ser Val Ala Leu Ala Thr 530Ala545Ser560Gly575Leu590Gln605Asp620Asn635Phe650Arg665Leu680Phe695Phe 7l0 Phe Leu Asn Met His Pro Thr Gly Ala Gln Arg Cys Asp Glu Gly Glu Lys Leu Asp Leu Val Leu Gly Val Pro Arg Glu His Pro Asp Met Ser Ser Tyr Val Ser Ala Ser GLys er Asp Thr Gly Gln Trp Ala Gly Glu 16 535Val550Thr565Pro580Gly595Lys610Ser625Gly640Arg655Arg670Gly685Gln 700 7l5 Leu Leu Glu Pro Ile Trp Ala Ala Leu Asp Ala Ala Cys Val Asn Tyr Arg Arg Leu Tyr Glu Glu Ala Lys Met Leu Gly Asn Phe Arg Gly Pro Glu Leu Phe Ala Asp Val Glu Ser Asn Lys Thr His Leu Cys Gln Ala 540Glu555Val570Ser585Ser600Ser615Arg630Leu645Phe660Gly675Gly690Ala705Ala 720 Arg721Arg736Ile751Val766Val781Gly796Glu811Pro826Pro841Arg856Pro871Glu886 Glu Asp Leu His Glu Arg Asp Ala Val Pro Gln Gln Gln Glu Ile Ser Val Asn Leu His Leu Ala Gly Ala Leu Gln Trp Phe Ala His Leu Ile Leu Val Trp Leu Leu Glu Lys Ser725Gln740Arg755Gln770Thr785Gly800Ser815Glu830Val845Thr 860 875Leu890 Trp Pro Ala Arg Ser Gly Val Arg Gln Arg Phe Leu Glu Phe Lys Glu Lys Ser Cys Val Leu Asp Phe Leu Ala Arg Arg Gly Met Lys Gln Pro Glu Glu Pro Leu Ser Leu Cys17 Ser Leu Phe Ser Glu Pro Asp Lys Arg Asp Thr Ser Pro Trp730Gln745Gln760Thr775Gly790Asn805Pro820Gly835Leu850Ile865Leu880Gln895 Thr Lys Leu Ala Leu Arg Pro Ser Pro Thr Ala Asp Leu Leu Thr Ala Gln Pro Ala Pro Pro Ser Glu Pro Leu Gln Pro Ile Thr Tyr Gly Pro Gly Cys Pro Glu Arg Glu Gly Arg Ile Gln Leu Thr Thr Pro Pro Arg Val Tyr735Leu750Ser765Leu780Pro795Val810Glu825Pro840Leu855Ser 870 885Tyr900 Leu 901Glu Gln916Gln Leu931 Pro Ser946Ala Tyr961 Cys Ser976Cys Phe991Ser Leu1006Phe Arg1021Gly Leu1036Ser Gln1051Gln Arg1066Pro Asp 1081 Phe Pro Thr Arg Thr Ile Pro Gly Gln Leu Gly Asn 905Pro Ser920Leu Leu935His Pro950 Thr Gln965Trp Leu980 Arg Gly995Cys Ile1010Phe Trp1025Pro Thr1040Asp His1055Val Phe1070Pro Lys 1085 Val Gln Gly Asp Ser Ala Leu Gln Pro Leu Gly Thr Ala Pro Glu Gly Gln Pro Val Glu Met Tyr Arg Tyr Leu Ile Leu Leu Ser Gly Arg Thr Arg Val Val 18 910Pro Ala925Tyr Tyr940His Leu955Gly Pro970Lys Pro985 Phe Arg1000Pro Gly1015 Leu His1030Leu Val1045Asp Glu1060Leu Thr1075Gln Asp 1090 Pro Ser Thr Leu Gly Leu Thr Asp Phe Val Ala Ile Leu Val Val His Asp Pro Gly Ile Gly Gln Phe Leu Leu Ser Ala Tyr Pro Pro Ile Glu Cys Asn Ser 915Leu Thr930Ser Ala945 Val Leu960Gly Val975 Val Pro990Asp Pro1005Ala Pro1020Ser Lys1035Arg Cys1050Ala Gln1065Arg Glu1080Thr Glu 1095 Leu Ala Ala Glu Val His Arg Val 1096 1100 Met Phe Val Cys Gly Asp Val Thr 1111 1115 Thr Val Gln Arg Ile Leu Ala Thr 1126 1130 Glu Ala Asp Val Ile Gly Val 1141 1145 His Glu Asp Ile Phe Gly Leu Thr 1156 1160 Ser Arg Ile Arg Thr Gln Ser Phe 1171 1175 Arg Gly Ala Val Pro Trp Ala Phe 1186 1190 Asn Ser Pro 1201 1203 To obtain polyclonal antibodies to NO synthase, it is also possible to use a Leu Cys1105Met Ala1120Glu Gly1135Leu Arg1150Leu Arg1165Ser Leu1180Asp Pro 1195 Leu Thr Asp Asp Thr Gln Pro Glu Asn Met Gln Gln Glu Gly Val Glu Gln Glu Arg Ser Gly His1110Leu Gln1125Leu Asp1140Arg Tyr1155Val Thr1170Gln Leu1185Asp Thr 1200 fragment of NO synthase, selected, for example, from the following sequences: SEQ ID NO: 3 Pro Trp Ala Phe 1192 1195 SEQ ID NO: 4 19 Gly Ala Val Pro 1189 1192 SEQ ID NO: 5 Arg 1185His Leu Arg Gly Ala Val Pro Trp Ala Phe Asp Pro Pro Gly Pro1186 1190 1195 1200Asp Thr Pro Gly Pro 1201 1205SEQ ID NO: 6Ala Phe Asp Pro Pro Gly Pro11941195 1200Asp Thr Pro Gly Pro1201 1205SEQ ID NO: 7His Leu Arg Gly Ala Val Pro Trp Ala Phe Asp1186 1190 11951196SEQ ID NO: 8 His Leu Arg Gly Ala Val Pro Trp Ala Phe Asp Pro Pro Gly Pro1186 1190 1195 1200Asp Thr Pro Gly Pro1201 1205 The exemplary procedure for preparation of the starting polyclonal antibodies toNO synthase may be described as follows. ln 7-9 days before blood sampling, 1-3intravenous injections of the desired antigen are made to the rabbits to increase thelevel of polyclonal antibodies in the rabbit blood stream. Upon immunization, bloodsamples are taken to test the antibody level. Typically, the maximum level of immunereaction of the soluble antigen is achieved within 40 to 60 days after the first injection of the antigen. Upon completion of the first immunization cycle, rabbits have a 30-day rehabilitation period, after which re-immunization is performed with another 1-3intravenous injections.

To obtain antiserum containing the desired antibodies, the immunized rabbits'Product clots formed on the tube sides are removed with a wooden spatula, and a rod is placed into blood is collected from rabbits and placed in a 50m| centrifuge tube. the clot in the tube center. The blood is then placed in a refrigerator for one night at thetemperature of about 40°C. On the following day, the clot on the spatula is removed, andthe remaining liquid is centrifuged for 10 min at 13,000 rotations per minute.Supernatant fluid is the target antiserum. The obtained antiserum is typically yellow.20% of NaN3 (weight concentration) is added in the antiserum to a final concentration of0.02% and stored before use in frozen state at the temperature of -20°C or without NaN3at the temperature of -70°C. To separate the target antibodies to gamma interferon fromthe antiserum, the following solid phase absorption sequence is suitable: 10 ml of the antiserum of rabbits is diluted twofold with 0.15 l\/I NaCl, after which6.26g Na2SO4 is added, mixed and incubated for 12-16 hours at 4°C. The sediment isremoved by centrifugation, diluted in 10ml of phosphate buffer and dialyzed against thesame buffer during one night at ambient temperature. After the sediment is removed, theThe antibody fraction is determined by measuring the optical density of the eluate at 280 nm. solution is applied to a DEAE-cellulose column balanced by phosphate buffer.

The isolated crude antibodies are purified using affine chromatography method byattaching the obtained antibodies to NO synthase located on the insoluble matrix of thechromatography media, with subsequent elution by concentrated aqueous salt solutions.

The resulting buffer solution is used as the initial solution for the homeopathicdilution process used to prepare the activated-potentiated form of the antibodies. Thepreferred concentration of the initial matrix solution of the antigen-purified polyclonalrabbit antibodies to NO synthase is 0.5 to 5.0 mg/ml, preferably, 2.0 to 3.0 mg/ml.

The polyclonal antibodies to endogenous biological molecule may also beobtained by a similar methodology to the methodology described for endothelial NO synthase antibodies using an adjuvant. 21 The resulting buffer solution is used as the initial solution for the homeopathicdilution process used to prepare the activated potentiated form of the antibodies.

The activated potentiated form of each component of the combination may beprepared from an initial solution by homeopathic potentization, preferably using themethod of proportional concentration decrease by serial dilution of 1 part of eachpreceding solution (beginning with the initial solution) in 9 parts (for decimal dilution), orin 99 parts (for centesimal dilution), or in 999 parts (for mi||esimal dilution) of a neutralsolvent, starting with a concentration of the initial solution of antibody in the solvent,preferably, water or a water-ethyl alcohol mixture, in the range from about 0.5 to about5.0 mg/ml, coupled with external impact. Preferably, the external impact involvesmultiple vertical shaking (dynamization) of each dilution. Preferably, separate containersare used for each subsequent dilution up to the required potency level, or the dilutionfactor.

"Homeopathic medicines", l\/l., 1967, p. 14-29, incorporated herein by reference for the This method is well-accepted in the homeopathic art. See, e.g. V. Schwabe purpose stated.

For example, to prepare a 12-centesimal dilution (denoted C12), one part of theinitial matrix solution of antibodies to NO synthase with the concentration of 3.0 mg/ml isdiluted in 99 parts of neutral aqueous or aqueous-alcohol solvent (preferably, 15%-ethylalcohol) and then vertically shaked many times (10 and more) to create the 1stcentesimal dilution (denoted as C1). The 2nd centesimal dilution (C2) is prepared fromthe 1st centesimal dilution C1. This procedure is repeated 11 times to prepare the 12thcentesimal dilution C12. Thus, the 12th centesimal dilution C12 represents a solutionobtained by 12 serial dilutions of one part of the initial matrix solution of antibodies withthe concentration of 3.0 mg/ml in 99 parts of a neutral solvent in different containers,which is equivalent to the centesimal homeopathic dilution C12. Similar procedures withthe relevant dilution factor are performed to obtain the desired dilutions.The intermediatedilutions may be tested in a desired biological model to check activity. The preferredactivated potentiated forms for antibodies comprising the combination of the invention isa C12, C30 and C200 dilutions for each activated-potentiated form. When using themixture of various homeopathic dilutions (primarily centesimal) of the active substance 22 as biologically active liquid component, each component of the composition (e.g., C12,C30, C50, C200) is prepared separately according to the above-described procedureuntil the next-to-last dilution is obtained (e.g., until C11, C29, and C199 respectively),and then one part of each component is added in one container according to the mixturecomposition and mixed with the required quantity of the solvent (e.g. with 97 parts forcentesimal dilution). lt is possible to use the active substance as mixture of various homeopathicdilutions, e.g. decimal and/or centesimal (D20, C30, C100 or C12, C30, C50 or C12,C30, C200, etc.), the efficiency of which is determined experimentally by testing thedilution in a suitable biological model, for example, in models described in the examplesherein. ln the course of potentiation and concentration decrease, the vertical shakingmay be substituted for external exposure to ultrasound, electromagnetic field or anysimilar external impact procedure accepted in the homeopathic art.

The solid unit dosage form of the pharmaceutical composition of the inventionmay be prepared by using impregnating a solid, pharmaceutically acceptable carrier withthe mixture of the activated potentiated form aqueous or aqueous-alcohol solutions ofactive components that are mixed, primarily in 1:1 ratio and used in liquid dosage form.Alternatively, the carrier may be impregnated consecutively with each requisite dilution.

Preferably, the pharmaceutical composition in the solid unit dosage form isprepared from granules of the pharmaceutically acceptable carrier which was previouslysaturated with the aqueous or aqueous-alcoholic dilutions of the activated potentiatedform of antibodies. The solid dosage form may be in any form known in thepharmaceutical art, including a tablet, a capsule, a lozenge, and others. As an inactivepharmaceutical ingredients one can use glucose, sucrose, maltose, amylum, isomaltose,isomalt and other mono- olygo- and polysaccharides used in manufacturing ofpharmaceuticals as well as technological mixtures of the above mentioned inactivepharmaceutical ingredients with other pharmaceutically acceptable excipients, forexample isomalt, crospovidone, sodium cyclamate, sodium saccharine, anhydrous citricacid etc), including lubricants, disintegrants, binders and coloring agents. The preferred 23 carriers are lactose and isomalt. The pharmaceutical dosage form may further includestandard pharmaceutical excipients, for example, microcrystalline cellulose, magnesiumstearate and citric acid.

The example of preparation of the solid unit dosage form is set forth below. Toprepare the solid oral form, 100-300 um granules of lactose are impregnated withaqueous or aqueous-alcoholic solutions of the activated potentiated form of antibodies tohistamine, activated-potentiated form of antibodies to NO synthase and the activatedpotentiated form of antibodies to an endogenous biological molecule in the ratio of 1 kgof antibody solution to 5 or 10 kg of lactose (1:5 to 1:10). To effect impregnation, thelactose granules are exposed to saturation irrigation in the fluidized boiling bed in aboiling bed plant (e.g. "Hüttlin Pilotlab" by Hüttlin GmbH) with subsequent drying viaheated air flow at a temperature below 40°C. The estimated quantity of the driedgranules (10 to 34 weight parts) saturated with the activated potentiated form ofantibodies is placed in the mixer, and mixed with 25 to 45 weight parts of “non-saturated” pure lactose (used for the purposes of cost reduction and simplification andacceleration of the technological process without decreasing the treatment efficiency),together with 0.1 to 1 weight parts of magnesium stearate, and 3 to 10 weight parts ofmicrocrystalline cellulose. The obtained tablet mass is uniformly mixed, and tableted bydirect dry pressing (e.g., in a Korsch - XL 400 tablet press) to form 150 to 500 mg roundpills, preferably, 300 mg. After tableting, 300 mg pills are obtained that are saturatedwith aqueous-alcohol solution (3.0-6.0 mg/pill) of the combination of the activated-potentiated form of antibodies. Each component of the combination used to impregnatethe carrier is in the form of a mixture of centesimal homeopathic dilutions, preferably,C12, C30 and C200.

While the invention is not limited to any specific theory, it is believed that theactivated-potentiated form of the antibodies described herein do not contain themolecular form of the antibody in an amount sufficient to have biological activityattributed to such molecular form. The biological activity of the combination drug(combination pharmaceutical composition) of the invention is amply demonstrated in the appended examples. 24 Preferably, for the purpose of treatment, the combination of the invention isadministered from once daily to four times daily, preferably twice daily, eachadministration including one or two combination unit dosage forms.

The invention is further illustrated with reference to the appended non-limiting examples.

EXAMPLESExample 1.

Study of effect of a complex preparation containing ultralow doses (ULD) ofactivated - potentiated forms of polyclonal affinity purified rabbit antibodies to brain-specific protein S-100 (anti-S100) and endothelial NO-synthase (anti-eNOS), obtainedby super-dilution of initial matrix solution (concentration: 2.5 mg/ml) (10012, 10000, 100200times), equivalent to a blend of centesimal homeopathic dilutions C12, C30, C200 (ratio:1:1) (ULD of anti-S100+anti-eNOS), as well as its components - activated - potentiatedform of polyclonal affinity purified rabbit antibodies to ultralow doses (ULD) of brain-specific protein S-100 (anti-S100), purified on antigen, obtained by super-dilution ofinitial matrix solution (10012, 10030, 100200 times, equivalent to a blend of centesimalhomeopathic dilution C12, C30, C200, and activated - potentiated form of polyclonalrabbit antibodies to ultralow dose of endothelial NO-synthase (ULD of anti-eNOS),obtained by sopar-dilution of initiai matrix solution (10012, 1oo3°, 100200 times), aquivaiantto a blend of centesimal homeopathic dilution C12, C30, C200 in vitro on binding ofstandard ligand [3H]pentazocine to human recombinant o1 receptor was evaluated usingradioligand method. Potentiated distilled water (blend of homeopathic dilutions C12+C30+C200) was used as test preparations control.

Sigma-1 (o1) receptor - an intracellular one which is localized in the cells of centralnervous system, the cells of the most of peripheral tissues and immune component cells.Receptors exhibit a unique ability to be translocated which is caused by manypsychotropic medications. The dynamics of sigma-1 receptors is directly linked tovarious influences which are performed by preparations acting to the sigma-1 receptors.

These effects include the regulation of activity channels, ecocytosis, signal transferring, remodeling of the plasma membrane (formation of rafts) and Iipid transportation /metabolism. All this can contribute to the plasticity of neurons in a brain. There isevidence that the sigma-1 receptors have a modulating effect on all the majorneuromediator systems: noradrenergic, serotonergic, dopaminergic, cholinergic systemsand Nl\/lDA- adjustable glutamate effects. Sigma-1 receptor plays an important role in(e-g.,Parkinson), psychiatric and affective disorders and stroke; and italso takes part in the the pathophysiology of neurodegenerative diseases Alzheimer's disease,processes of learning and memory. ln this regard, the ability of drugs to influence theefficiency of interaction of ligands with sigma-1 receptor indicates on the presence ofneuroprotective, anti-ischemic, anxiolytic, antidepressant and anti astenic components inthe spectrum of its pharmacological activity that allows the consideration of these drugs as effective preparations particularly for the treatment of cerebrovascular diseases.

During the test (to measure total binding) 20 ul of complex preparation of ULD ofanti-S100+anti-eNOS or 10 ul of ULD of AB to S100 or 10 ul of ULD of AB to NOS weretransferred in the incubation medium. Thus, the quantity of ULD of anti-S100+anti-eNOS, transferred into the test basin when testing the complex preparation was identicalto that of ULD of AB to S100 and ULD of AB to NOS tested as monopreparations, whichallow comparing the efficiency of the preparation to its separate components. 20 ul and10 ul of potentiated water were transferred in the incubation medium.

Further, 160 ul (about 200ug of protein) ofJurkat cell line membranes homogenate(human leukemic T-lymphocyte line), and finally, 20 ul of tritium-labeled radioligand [3H]pentazocine (15 nm) were transferred. ln order to measure non-specific binding, 20 ul of non-labeled ligand- haloperidol(10 ul\/l) were transferred in the incubation medium instead of the preparations orpotentiated water.

Radioactivity was measured using a scintillometer (Topcount, Packard) andscintillation blend (l\/licroscint 0, Packard) following the incubation within 120 minutes at22°C in 50 mM Tris-HCI buffer (pH = 7,4) and filtration using fiberglass filters (GF/B, 26 Packard). Specific binding (during the test or control) was calculated as a differencebetween total (during the test or control) and non-specific binding.

Results are represented as percentage of specific binding inhibition in control5 (distilled waterwas used as control) (Table 1).

Table 1.% of radioligand specific binding in % ofQuantity control radioligandTest group per test St nd bindingbasin 1 test 2 test Average inhibition incontrol ULD of anti- 58,0S100+anti- 20 ul eNOS 48,4 35,5 42,0 ULD of anti- 10 l 34,8 s1oo l* 67,3 63,1 65,2 ULD of anti- 10 | -54,3eNos i* 147,5 161,1 154,3 Potentiated 20 | 13,1 water l* 96,1 75,6 66,9 Potentiated 10 | -23,2water i* 140,1 106,2 123,2 10 Effect of the preparations and potentiated water on binding of standard ligand [3H]pentazocine to human recombinant o 1 receptor Note: % of specific bindinq in control = (specific binding during the test/ specificbinding in control)* 100%; % of specific bindinq inhibition in control = 100% - (specific binding during the 15 test/ specific binding in control) * 100%). 27 The outcomes reflecting inhibition above 50% represent significant effects of thetested compounds; inhibition from 25% to 50% confirm mild to moderate effects;inhibition less than 25% is considered to be insignificant effect of the tested compound and is within background level.

Therefore, the conditions of this test model showed that the complex preparationof ULD of anti-S100+anti-eNOS is more efficient than its separate components (ULD ofanti-S100 and ULD of anti-eNOS) in inhibiting the binding of standard radioligand[3H]pentazocine to human recombinant o1 receptor; ULD of anti-S100, transferred intothe test basin, namely 10 pl, inhibit the binding of standard radioligand [3H]pentazocineto human recombinant o1 receptor, but the effect intensity is inferior to that of thecomplex preparation of ULD of anti-S100+anti-eNOS; ULD of anti-eNOS, transferredinto the test basin, namely 10 pl, had no effect on the binding of standard radioligand[3H]pentazocine to human recombinant o1 receptor; potentiated water, transferred intothe test basin, namely 10 pl or 20 pl, had no effect on the binding of standard radioligand[3H]pentazocine to human recombinant o1 receptor.

Example 2.

Alzheimer's disease (AD) is a neurodegenerative disease that is characterized bylowering of cognitive functions, memory deterioration, confused consciousness, andemotional changes. Although the main cause of this pathology is nowadays consideredthe accumulation of beta amyloid which leads to the formation of beta-amyloid plaquesand neurofibrillary tangles in brain tissues; AD is also accompanied by a deficiency ofcholinergic system. This is the basis of a most common way of modeling of AD inanimals with the help of antagonist of cholinergic system of scopolamine. Injection ofscopolamine into experimental animals (usually rats or mice) interrupts the ability tolearn and leads to deterioration of memory.

Various methods were used to assess cognitive functions of rats and mice, including l\/lorris water maze. The essence of this test is that the animals are released28 into a container with cloudy water from different points are forced to look for a hiddenfixed platform. The advantage of this method is that it allows the researcher to monitorthe process of animal training (the formation of ideas about the spatial alignment of theplatform no matter where the animal was placed in the water) so as to assess thememory strength (for this the test is conducted when the platform is removed).

The effectiveness in rats with Scopolamine amnesia of the combinationpharmaceutical composition of the present invention containing activated-potentiatedforms of polyclonal affinity purified on antigen of rabbit brain-specific proteins S-100(anti-S100) and to endothelial NO-syntheses (anti-eNOS) in ultra low doses (ULD)obtained by super dilution of storage stock solution (with concentration of 2.5 mg/ml) in10012, 10030, 100200 times, equivalent to centesimal homeopathic dilutions C12, C30,C200 (ULD anti-S100 + anti-eNOS) is studied. ln a study of the effectiveness of the drug ULD anti-S100 + anti-eNOS in rats withscopolamine amnesia (a model of Alzheimer's disease) 48 male rats of the Rj: Wistar(Han) line (weight 180-280g) were used. During 4 days the rats were subdermallyinjecting with normal saline (n = 12, intact) or scopolamine in doze of 0.5 mg / kg (n =36) (scopolamine-induced amnesia). Rats with scopolamine-induced amnesia weredivided into three groups and administered with distilled water (7.5 ml / kg, n = 12,control group 1), or ULD anti-S100 (7,5 ml / kg, n = 12, group 2) or ULD anti-S100 +anti-eNOS (7,5 ml / kg, n = 12, group 3) intragastrically for 9 days (4 days prior to theinjection of scopolamine, 4 days against the background of scopolamine and 1 day afterthe last scopolamine injection).

The training session in the l\/lorris water maze was conducted within 4 days of thescopolamine injection through 60 minutes after administration of tested drugs and 30minutes after administration of scopolamine (4 sequential tests at interval of 60seconds). Morris' maze is a round reservoir (diameter - 150 cm, height - 45 cm) at 30cm filled with water (26-28 ° C). At 18 cm from the edge of the container there is hiddenplatform (diameter - 15 cm) buried on 1.5 cm below the water level. Cloudy water madeby adding a non-toxic dye (e.g., milk powder) makes the platform invisible. For each test 29 the animal was placed in a maze in one of the initial points that are equidistant from thehidden platform and the animal was allowed to find the platform. lf the animal could notfind the platform within 120 seconds, the animal was put on the platform and left for 60seconds and the test was restarted. During the four tests in random order the animalsbegan to walk through the maze twice from each starting point. The tests were recordedon videotape and then analyzed for distance overcomes searching the platform in eachtrial and the latent period of searching for the platform. On day 5 the test wasperformed: the platform was removed from the maze and rats were given free float for 60 seconds. The time spent in the place where the platform used to be was recorded.

The administration of scopolamine significantly worsened the ability of animals tolearn. ln the control group the time spent by animals searching for platforms and thedistance that animals swam searching for the platform, significantly increased (Table 2,3). The test shows that the memory of animals in the control group worsened: theanimals in this group spent less time in the place where the platform used to be locatedThe administration of ULD anti-S100 didn't lead to improvement of the studied parameters (Tables 2, 3, 4). The administration of ULD anti- than intact animals (Table 4).

S100 + anti-eNOS led to some improvement in learning which resulted in a shortening ofthe latent time of the platform search time (Table 2) and covered distance (Table 3)within 4 days of training and an improvement of memory as reflected in increase of the time spent in a place where the platform used to be located (Table 4).

Table 2.

Latent period of the platform search, sec Group Training 1st day 2"” day 3” day 4"' day lntact, n=12 54.7i6.2 30.8i2.8 26.9i5.'| 20.5i3.6 Control, n=12 100.116.8*** 92.419.3*** 81.4110.7*** 77.719.4***ULD anti-S100, n=12 106.817.0 99.317.8 95.619.0 80.4111.1ULD anti-S100 + anti- 94.417.2 90.718.2 78.318.6 60.1110.2eNOS, n=12*** - difference from intact is significant, p<0.05Table 3.Distance overcome to search the platform, cmGroup Training 1st day 2"” day 3” day 4m daylntact, n=12 1055.7194.6 659.5162.2 564.81119.3 406.1161.2Control, n=12 2587.11217.2*** 2559.61250.5*** 2397.91312.6 2366.11293.8***ULD anti- 2797212089 2865.21255.1 2857.01300.8 2457.41344.4S100, n=12ULD anti-S100 2434.31222.8 2529.91282.7 2344212830 1905.11343.7+ anti-eNOS,n=12 *** - difference from intact is significant, p<0.05 Table 4.

Time spent in a place where the platform used to be located, sec.

Group Test0-30 sec. 30-60 sec. 0-60 sec.lntact, n=12 40.814.1 36.813.6 38.512.6 31 control, n=12 18.412.8*** 18.811 .9*** 18.811 .7***ULD anﬁ-s1oo, n=12 18.812.1 21 _5128 17.811 .3:tå antfsmo J' amFeNOS' 19.114.8 238122 21.212.5 *** - difference from intact is significant, p<0.05 Thus, in model of Alzheimer's disease, the administratoin of the complex ULDanti-S100 + anti-eNOS was more effective in comparison with administration of ULDanti-S100 and vehicle.

Example 3.

The preclinical research studied the ultra low doses (ULD) of activated -potentiated forms of polyclonal affinity purified rabbit antibodies to brain-specific proteinS-100 (anti-S100) purified on antigen, and endothelial NO-synthase (anti-eNOS),obtained by super-dilution of initial matrix solution (concentration: 2,5 mg/ml) (10012,10030, 100200 times), equivalent to a mixture of centesimal homeopathic dilutions C12,C30, C200 (ratio: 1:1) (ULD anti-S100+anti-eNOS) in treating ischemic stroke caused by prefrontal cerebrocortical photothrombosis in rats.

Acute cerebrovascular disease (brain stroke) ranks third among Iethality causesin developed countries and one of the main causes of disability in humans (Gusev E.l.,2003; Janardhan V., QureshiAJ., 2004).

The photo-induced thrombosis model meets almost all requirements to theexperimental model of focal cerebral ischemia. The method developed by Watson(Watson B. et al., 1985) is based on the effect of light with wavelength 560 nm onphotosensitive pigment Bengal rose introduced into the blood flow. Active oxygen formsare created and caused increase in adhesiveness of endothelium cells and platelets, and formation of clots closing vascular lumens. The method of ischemic brain lesion 32 induction by using photo-induced thrombosis is technically simple and to close to clinicalforms of ischemic brain stroke. A great advantage of this model is that it is non-invasive,i.e. does not require craniotomy and, therefore, more accurately reproduces clinical picture of cerebral thrombosis.

Thirty seven male Wistar rats (weight: 150-180 g; age: 2-3 months) were includedin the study of the activity of ULD anti-S100+anti-eNOS in rats with ischemic strokecaused by prefrontal cerebrocortical photothrombosis. Bilateral focal ischemic injury inprefrontal cerebral cortex in rats was induced using the photochemical thrombosismethod by Watson (Watson B. D. et al., 1985) as modified by l.V. Viktorov (RomanovaG.A. et al, 1998). Bengal rose (3% solution) was injected in the jugular vein ofanesthetized rats (n=37) (anesthesia: chloral hydrate 300 mg/kg, intraperitoneally).Using a fiber optic bundle (3 cm in diameter) the light beam from halogen lamp (24 V,250 W) was delivered to the skull surface above the frontal cortex of the left and rightcerebral hemispheres to induce photothrombosis. Sham-operated rats (n=6) weresubject to the same procedure except administration of bengal rose and exposure tohalogen lamp light. The intact group included 6 rats.

Five days before and 9 days after stoke induction the following preparations wereadministered to rats with photothrombosis: distilled water (control-photothrombosis, 5ml/kg daily, n=12), ULD anti-S100 (5 ml/kg daily, n=7) or ULD anti-S100+anti-eNOS (5ml/kg daily, n=6). On Day 8 after the operation (or sham operation) conditioned passiveavoidance reflex (CPAR) test was performed to assess learning capability and memoryin rats. Rats were placed in a unit consisting of illuminated site and connected darkchamber, where animals were exposed to electric foot-shock of 0,45 mA due to whichusually preferred dark chamber became dangerous. Development of conditionedpassive avoidance reflex was tested on the next day. At that, rats were placed in theilluminated chamber. Latent period of the first entry in the dark chamber was recorded. lfa rat avoided the dark chamber for a long time, a conclusion was made that itremembered the danger (electric shock). The longer the latent period of entry in the darkchamber, the better the memory. 33 Volume of the stroke lesion was morphologically assessed in a proportion of ratsof experimental groups on Day 9. ln control rats photothrombosis caused formation of a large stroke area and, 5 therefore, leaded to memory impairment: CPAR reproduction worsened by 9.6%compared to intact rats and by 22.9% compared to sham-operated (Table 5).Administration of ULD anti-S100 reduced the stroke volume by 42.2% and improvedmemory by 14.0% compared to control-photothrombosis group. Administration of ULDanti-S100+anti-eNOS was more effective: the stroke volume reduced by 44.0% , and 10 conditioned reflex reproduction - by 33.4% compared to control-photothrombosis group.

Therefore, administration of the complex preparation of ULD anti-S100 + anti- eNOS was more efficient than monocomponent preparation of ULD anti-S100.

Table 5.

Volume of focal Latent period of CPARstroke (mm3); the (seconds), the number number of animals of animals lntact - 135.8 i 28.8; n=6Sham-operated - 159.3 i 18.7; n=6Control-photothrombosis 3.41 i 0.5; n=9 122.8 i 20.9; n=12Photothrombosis + ULD anti-S100 1.97 i 0.6; n=4 140.0 i 26.5; n=7Photothrombosis + ULD anti- 1.91 i 0.5; n=4 163.8 i 16.2; n=6 S100+anti-eNOS 15 Example 4.Study of the combination of “activated” potentiated forms of antibodies to a C-terminal fragment of the angiotensin ll AT1-receptor, in a mixture of homeopathic 34 dilutions of C12, C30, C200, with the activated potentiated form of antibodies toendothelial NO-synthase, in a mixture of homeopathic dilutions of C12, C30, C200, inSHR rats in a model of hypertension.

The combination of the “activated” potentiated form of antibodies to a C-terminalfragment of the angiotensin ll AT1-receptor, in a mixture of homeopathic dilutions ofC12, C30, C200, and the activated potentiated form of antibodies to endothelial NO-synthase in a mixture of homeopathic dilutions of C12, C30, C200, was studied, insolution form, in the SHR rat hypertension model. investigations were conducted on 40SHR line male rats from (weight 350i50 g, age 4.5 - 5 months) with hypertension, whichwere divided into 4 groups of 10 animals each.

For 28 days, the animals were treated as follows. Group 1 - 2.5 ml/kg of thepotentiated activated form of antibodies to a C-terminal fragment AT1 of humanangiotensin ll receptor (a mixture of aqueous dilutions C12, C30, C200) in combinationwith 2.5 ml/kg of distilled water, Group 2 - 2.5 ml/kg of the potentiated activated form ofantibodies to endothelial NO-synthase (a mixture of aqueous dilutions C12, C30, C200)in combination with 2.5 ml/kg of distilled water, Group 3 - 5 ml/kg of the combinationpharmaceutical composition (a mixture of aqueous dilutions C12, C30, C200 for eachcomponent), and Group 4 - 5 ml/kg of distilled water.

Systolic blood pressure (SBP) of awake rats was measured with the aid of anindirect method in a tail artery (using a cuff) once a week and 9 hours after the lastadministration of medicines.

All tested compositions demonstrated hypotensive effect (p<0.05): by 28th day,systolic blood pressure (SBD) decreased in comparison with the initial level in Group 1by - 20.6%; in Group 2 by 14.4%; in Group 3 by 27.6%. ln the control Group 4, SBDchanges were 1.6% in comparison with the initial values. The results demonstrate a clear synergistic hypotensive effect of the combination pharmaceutical composition.

Example 5.Study of the combination of the activated potentiated forms of antibodies to a C- terminal fragment of angiotensin ll AT1-receptor, in a mixture of homeopathic dilutions of C12, C30, C200, with the activated potentiated form of antibodies to endothelial NO-synthase, in a mixture of homeopathic dilutions of C12, C30, C200, in NISAG rats in amodel of hypertension.

The combination of the activated potentiated form of antibodies to a C-terminalfragment of angiotensin ll AT1-receptor, in a mixture of homeopathic dilutions of C12,C30, C200, and the activated potentiated form of antibodies to endothelial NO-synthasein a mixture of homeopathic dilutions of C12, C30, C200, was studied, in solution form,in the NISAG rat hypertension model. lnvestigations were conducted on 50 NISAG linemale rats (weight 300 g, age 4 months) with hereditary stipulated stress-sensitive arterialhypertension, which were divided into 5 groups by 10 animals each.

The animals were given per orally, once a day and for 28 days, the followingmedications: Group 1 - 2.5 ml/kg of the activated potentiated form of antibodies to a C-terminal fragment AT1 of human angiotensin ll 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 theactivated potentiated form of antibodies to endothelial NO-synthase (a mixture ofdilutions C12, C30, C200) in combination with 2.5 ml/kg of distilled water; Group 3 - 5ml/kg of the combination pharmaceutical composition (a mixture of homeopathicaqueous dilutions C12, C30, C200 of each component); Group 4 - 5 ml/kg (10 ml/kgdose) of the comparison drug (losartan); and Group 5 - 5 ml/kg of distilled water.

Two times a week, 2 to 6 hours after administration of VSD antibodies andlosartan, systolic blood pressure (SBP) was measured by an indirect method in a tailartery (using a cuff). The Table 6 shows the dynamics of changes in systolic bloodpressure in NISAG line rats, measured by indirect method.

Table 6.lndicator Initial SBP in SBP after 28 A in % of themmHg days of comparison initial levelmedicine with the initialadministration level, inin mmHg mmHgVSD antibodies to C- 176 150 -26 -14.7%terminal fragment AT1 of 36 5 human angiotensin llreceptor VSD antibodies toendothelial NO-synthase 175 164.5 -10.5 -6% Combination medicine on 179.5 140 -39.5 -22%the basis of VSDantibodies to C-tailedfragment AT1 ofangiotensin ll receptorand to endothelial NO- synthase Losartan 173.5 140.5 -33 -19% Control (distilled water) 181 178 -3 -1.6% Example 6.

The experimental studiy investigated the effects of antibodies to the C-terminalfragment to the insulin receptor ß-subunit affinity purified on antigen, in ultra-low dose,obtained by super diiutieri ef the iriitiai matrix eeiutieri 10012, 10000, 100200 times (ULDanti-IR), antibodies to endothelial NO-synthase affinity purified on antigen, in ultra-lowdeee, obtained by riyper-diititieri ef the iriitiai matrix eeititieri 10012, 10000, 100200 (ULDanti-ULD anti-eNOS), as well as the combination of ultra-low doses of antibodies to theC-terminal fragment to the insulin receptor ß-subunit and ultra-low dose of antibodies toendothelial NO-synthase (ULD anti-IR + ULD anti-eNOS). ln the study, 150 male Wistar were used (weight at beginning of study 250-300 g,age 3.5-4 months). 10 rats were intact. The rest were intravenously injected withstreptozotocin at the dose of 50 mg/kg (experimental model of diabetes mellitus). 72hours after injection of streptozotocin, rats with blood plasma glucose level not less than12 mmol/l were selected, divided into 7 groups (20 rats in each), which over 21 daysinsulin® (8 units/kg/day, subcutaneously), Rosiglitazone® (8 mg/kg/day, twice daily intragastrically), were given distilled water (5 ml/kg/day, once daily intragastrically), ULD anti-IR (2.5 ml/kg/day in a volume of 5 ml/kg/day, once daily intragastrically), ULDanti-IR + ULD anti-eNOS Rosiglitazone® and insulin® together or ULD anti-IR + ULD anti-eNOS and insulin®,37 (5 ml/kg/day, once daily intragastrically), and also according to regimes corresponding to each preparation (as described above). lntactrats received distilled water in the same volume. On days 7, 14 and 21 of injection ofpreparations in rats, fasting blood plasma glucose level measured with enzymaticmethod (glucose oxidase method) with utilization of “glucose FKD” kits (Russia).

Oral glucose tolerance test (OGTT) was performed on day 14 of the study (day 14of administration of preparation) according to standard method (Du Vigneaud and Karr,1925). The rats were starving at water for 18 hours. 60 min before the test they were lastgiven test substances. lntact rats received distilled water in the same volume. Glucosewas administered per os 50% w/w water glucose solution (1 g/kg of rat weight). Serumglucose of blood sample from tail vein was measured by using “Glucose FKD” kit (OOO"Pharamaceutical and clinical diagnostics, Russia, www.fkd.ru) at 0, 30, 60, 90, 120 min.l\/lean area under the curve (AUC) concentration of blood glucose over time wascalculated.

Injection of streptozotocin led to a substantial increase in blood plasma glucose ofrats in comparison with intact animals (18 mmol/l versus 3.5 mmol/l, p<0.05). ln the ULDanti-IR group, on day 7, 14 and 21 of injection of preparation, glucose level was lowerthan in the control group by 22-28% on average; however, differences did not reach astatistically significant level. The combination of ULD anti-IR and anti-eNOS was moreefficacious; the decrease in glucose level on days 14 and 21 of the experiment were47% and 42%, respectively (p<0.05 versus control). The reference preparation,Rosiglitazone, also lowered glucose level by day 14 and 21 of the experiment; at that,the effect reached statistical significance on day 14 of the experiment only (36%, p<0.05versus control).

Insulin, injected at 1/2 of the effective dose (selected in the preliminary study) mosteffectively lowered glucose level in all observation periods (down to the level of the intactcontrol). (Figure 1). lt should be taken into account that short-acting insulin was used inthe study and blood plasma glucose was measured 1 hour after its injection, which alsoinfluenced the effect of the 1/2 insulin dose on blood glucose level. Against thisbackground it was not possible to fully determine what the effect of the combined use of insulin and rosiglitazone or insulin and complex ULD anti-IR + anti-eNOS is. 38 Glucose tolerance disturbance (reduction in glucose utilization by the body) is oneof the most important indicators in diagnostic and treatment of diabetes mellitus. ln intactanimals, in the oral glucose tolerance test (day 14 of injection of preparations), complexpreparation ULD anti-IR + ULD anti-eNOS and insulin most effectively increased glucosetolerance when administered alone. Rosiglitazone also reduced the area underconcentration over time curve (increased glucose tolerance); however, its efficacy was not statistically significant versus the control group (Figure 2).

Example 7.

The experimental studiy investigated the effects of antibodies to the C-terminalfragment to the insulin receptor ß-subunit affinity purified on antigen, in ultra-low dose,obtained oy super oiiution of tne initiai matrix eoiution 10012, 10000, 100200 times (ULDanti-IR), antibodies to endothelial NO-synthase affinity purified on antigen, in ultra-lowoioee, ootaineoi oy nyper-oiiution of tne initiai matrix eoiution 10012, 10000, 100200 (uLDanti-ULD anti-eNOS), as well as the combination of ultra-low doses of antibodies to theC-terminal fragment to the insulin receptor ß-subunit and ultra-low dose of antibodies toendothelial NO-synthase (ULD anti-IR + ULD anti-eNOS). ln the study, 36 male Goto-Kakizaki rats were used (weight at beginning of study250-280 g, age 10-12 weeks). Rats of this line are characterized by spontaneousdevelopment of non-insulin-dependent diabetes. The animals were divided into 3 groups(12 rats in each) and received either distilled water (5 ml/kg, once daily intragastrically),or ULD anti-IR (2.5 ml/kg once daily intragastrically), or ULD anti-IR + ULD anti-eNOS (5ml/kg, once daily intragastrically) for 28 days. Blood plasma glucose level was measuredwith the help of a glucose analyzer (Beckman, Fullerton, California, USA) beforebeginning injection of preparations and on day 4, 8, 12, 16, 20, 24, 28 of injection ofpreparations. On day 28, a glucose tolerance test was carried out (glucose p.o., 1 g/kg).

Injection of ULD anti-IR led to a significant (p<0.05) drop in blood plasma glucoselevel of rats; however, the use of complex ULD anti-IR + ULD anti-eNOS was more efficacious (p<0.001 versus control) (Figure 3). 39 The results were confirmed by glucose tolerance test data carried out on day 28of injection of preparations (Figure 4). Injection of ULD anti-IR led to an increase inglucose tolerance (statistically insignificant drop by 44% AUC versus control). At thesame time, the reduction in this parameter (AUC) caused by injection of complex ULDanti-IR + ULD anti-eNOS was 62% and it was statistically significant versus control(p<0.05).

Example 8.

The following preparation were used: 300 mg tablets impregnated with aqueousalcoholic solution (3 mg / tab.) activated - potentiated form of polyclonal rabbit brain-specific proteins antibodies S-100, purified on an antigen, in ultra low dose (ULD anti-S100) received by super dilution of initial solution (with concentration of 2.5 mg / ml) in10012, 10030, 100200 times, of equivalent mixture of centesimal homeopathic dilutionsC12, C30, C200; 300 mg tablets impregnated with pharmaceutical compositioncontained aqueous-alcohol solutions of (6 mg/tab) activated - potentiated forms ofpolyclonal affinity purified rabbit antibodies to brain-specific protein S-100 (anti S-100)and to eNOS (anti-eNOS) in ultra low dose (ULD), received by super dilution of initialsolution (with concentration of 2.5 mg/ml) in 10012, 10030, 100200 times, of equivalentmixture of centesimal homeopathic dilutions C12, C30, C200; (ULD anti-S100+anti-eNOS); 300 mg tablets impregnated with aqueous-alcohol solution (3 mg/tab.) ofactivated - potentiated form of polyclonal rabbit eNOS antibodies purified on antigen inultra low dose (ULD anti-eNOS), received by super dilution of initial solution (withconcentration of 2.5 mg/ml) in 10012, 10030, 100200 times, of equivalent mixture ofcentesimal homeopathic dilutions C12, C30, C200; and as placebo 300 mg tabletscontaining excipients: lactose (lactose monohydrate) - 267 mg, microcrystal cellulose -30 mg, magnesium stearate - 3 mg.

The effectiveness of the studied drugs in the treatment of dizziness (vertigo) andother symptoms of motion sickness was evaluated on kinetosis model or motion diseases/motion sicknesses which occurs by various vestibular vegetative disorders.

Dizziness is the typical sign of lesion of the vestibular analyzer of various genesisincluding dysfunction of the vestibular nerve and cochlear system, circulatoryembarrassment in vertebrai basiiar system, pathology of the central nervous system(CNS), etc. Dizziness as a manifestation of kinetosis accompanied with other vestibular-vegetative disorders which include three types of reactions: the vestibular-motor(nystagmus and the reaction of deviation), vestibular-sensory (in addition to dizziness,nystagmus is (or reaction of post rotation), defensive movements) and vegetative(nausea, vomiting, sweating, palpitation, heat feeling, pulse and blood pressurefluctuations).

Double blind placebo controlled comparative study were conducted in parallelgroups consisting of 15 somatically healthy subjects - males and females aged from 15to 60 years (mean age 33.3*_fO.75 years) with low (n=5; 33%) or mean (n=10; 67%)degree of motion sickness resistance in order to test anti motion sickness properties ofvarious compositions. Group I was given ULD anti-S100+anti-eNOS, Group 2 was givenULD anti-S1OO and Group 3 was given anti-eNOS.

To simulate the condition of motion sickness and evaluate the effectiveness ofstudied drugs the most appropriate and recognized kinetosis models - test with acontinuous cumulative effect of accelerations by Coriolis (CCEAC) was used. Initialtolerance of CCEAC test in all study subjects was not more than 5 minutes. Vestibular-vegetative disorders provoked by kinetic effect (CCEAC) were registered with usage ofcomplex of diagnostic methods including subject's examination, quantitative evaluationof disorders of vestibular-vegetative sensitivity (Halle scale), analysis of heart ratevariability (HRV), and self-esteem of functional condition (WBAM - well-being, activity,and mood). As the criteria of efficiency of conducted therapy the dynamics of toleranceand extent of recovery period at kinetic influence were assessed as well as alteration ofindexes' evidence of sensory-motor reactions (nystagmus), HRV indexes (with usage ofBiocom Wellness Scan system, developed by AWS, LLC in accordance withInternational Standard of European Cardiologists Association and North AmericanElectrophysiology Association) and WBAl\/I data. The safety criteria were character, evidence and terms of emergence of probable adverse events (AE) in the treatment41 period connected with medication intake; influence of studied drugs for indexes whichcharacterize the function of central nervous system (CNS) (reaction on moving object(Rl\/lO)), the time of simple motor reaction (TSl\/lR); the dynamics of physical andfunctional factors (heart rate (HR), systolic and diastolic blood pressure (SBP, DBP),Stange's test; exercise tolerance (index of Harvard step-test). Safety was assessed aftersingle dose administration and after 7-day course administration of the combination ULDanti-S-1OO and ULD anti-eNOS.

All the subjects during 1 month before being involved into the study had not takenany drugs. After screening the subjects were randomized into 4 groups (Group1 - ULDanti-S100+anti-eNOS, Group 2 - ULD anti-S100, Group 3 - ULD anti-eNOS, and Group4 - placebo).

On the first day of the study (Visit 1) the initial functional and psycho-physiologicalstate of the subjects was registered, the subjects were then given 5 tablets of therespective ULD antibodies, followed by administration of the CCEAC test. The durationof the test was registered; vegetative-vestibular disorders and AEs related to motionsickness were detected with the help of a complex diagnostic examination. ln the next2-6 days the subject were given 1 tablet three times a day of the prescribed drug. At the7"' day (Visit 2) the subjects were given the same dosage as on the first day (Visit 1).The complex of diagnostic studies was conducted before and after the CCEAC test.The study was organized in such way as study crew would work only with one subject.The study was parallel and conducted in the first half of a day with participation of, as arule, 4 persons in a day, one person for drug or placebo. The next three weeks werewashout period, at the end of which the new drug or placebo was prescribed to subjectsof each group; the cycle of study was being repeated (Visit 1, the course intake of adrug; Visit 2). Thus, during the study each subject took part in four cycles of study. Thatis, each subject participated in each group with a three-week washout period betweeneach cycle. This allowed the researcher to level the influence of individual peculiaritiesof a test person on the treatment effect. The analysis of drug efficiency was conductedon the data of all the test subjects who has completed the full course of studied drug intake according to study protocol (n=15).42 The evidence factors of symptoms of motion sickness (vertigo, nausea, inactivity,skin pallor, sweatiness, etc.) after kinetic influence (CCEAC) against the background ofsingle-day intake of studied drugs evidenced that all the study subjects have gainedroughly the same state of motion sickness as far as the evidence of assessed symptomsof vegetative dysfunction on Halle's scale by physician-researcher was not differedsignificantly in all groups (Table 7, Visit 1). However, while the kinetic affect whichcause similar symptoms of motion sickness was different in four groups and wasdependent on the drug which was taken by the subjects of the study (Table 8, Visit 1).One-day intake of ULD anti-S100 + anti-eNOS preparation led to most clearly antimotion sickness effect which manifested itself not only in significantly more time oftolerance of CCEAC test (104.10 i 13.14 sec. vs. 68.50 i 6.57 sec. - in the group ofULD anti-S100; 75.00 i 6.79 sec. - in the group of ULD anti-eNOS and 61.30 i 3.15sec. - in the placebo group) but also in the least time of nystagmus (9.90 i 1.20 sec. vs.13.50 i 1.51; 16.10 i 1.68 and 13.30 i 1.12 sec., respectively) and in maximal rapidrecovery (9690 i 13.54 sec. vs. 194.20 i 18.45; 202.50 i 21.72 and 241.70 i 38.41 sec., respectively).

Roughly similar indexes were registered at Visit 2 after receiving a course ofdrugs. To achieve the similar symptoms of motion sickness (Table 7, Visit 2) the longesttime of kinetic impact was applied to the subjects who has been receiving thecomposition of ULD anti-S100 + anti-eNOS (Table 8, Visit 2) for 7 days. pronounced anti motion sickness effect of the composition of ULD anti-S100 + anti- The most eNOS was expressed in significantly less time of nystagmus (9,50 i 1,38 sec, p <0.01)and duration of the recovery period (117.90 i 15.65 sec; p <0.01). The monocomponentpreparation ULD anti-S100 had anti motion sickness action as better indexes oftolerance of CCEAC test, recovery time of nystagmus and recovery than in the placebogroup evidenced (Table 8, Visits 1 and 2), but the efficacy of ULD anti-S100 was inferiorto composition of ULD anti-S100 + anti-eNOS. The monocomponent preparation ULDanti-eNOS did not show anti motion sickness effect since the results of CCEAC testsand subsequent recovery period had no significant difference from the placebo group (Table 8, Visits 1 and 2). Comparative analysis of indexes of CCEAC test in the groups 43 of ULD anti-S100 + anti-eNOS and ULD anti-S100 in one-day intake of the drugs hasshown that the addition of ULD anti-eNOS increased the tolerance of the kinetic effecton the 52%, reduced the nystagmus time on 27% and contributed to the reduction therecovery period after the end of the kinetic effect on 50% including the duration ofdizziness - on 49%. However, the greatest contribution of the component of ULD anti-eNOS introduced the effectiveness of combined preparation (compositions of ULD anti-S1OO + anti-eNOS) in course intake of a drug which was expressed in excess of 30% ofthe result achieved in the group of ULD anti-S100 by factors of tolerance of kinetic effectand nystagmus duration (in each of the parameters). ln addition, the growth of the effecton Visit 2 by indexes of toierance of CCEAC test and duration of the nystagmus inrelation to data of Visit 1 when taking the composition ULD anti-S100 + anti-eNOS incomparison to monocomponent preparation ULD anti-S100 was expressed in a greaterdegree as confirmed by alteration of these indexes on 30% and 4% (versus 21% and0% in the ULD anti-S100 group). ln assessing the effectiveness of anti motion sicknessproperties of drugs the special attention was paid to the possible impact of drugs on thestability of autonomic nervous system (ANS) in particular, shifting of the balancebetween its sympathetic and parasympathetic divisions. For this purpose, at each visitHRV parameters were analyzed at the rest condition and when performing the functionaltests (breathing and orthostatic tests).

Table 7 indexes of Ha||e's scale depending on applied preparation after theperformance of CCEAC test Ha||e's scale (points) Preparation Visit 1 Visit 2 (one-day intake) (course intake) (n=15; IVIiSE) (n=15; IVIiSE) ULD anti-S100 + anti-eNOS 12.00i0.63 12.30i0.59 44 ULD anti-S100 13.3010.65 12.3010.46 ULD anti-eNOS 13.1010.78 12.0010.55 Placebo 13.4010.77 13.3010.45Table 8 The dynamics of indexes of CCEAC test depending on applied preparation Visit 1 (one-day intake) Preparati0n Tolerance of Nystagmus Recovery time, sec.CCEAC test, sec. time, sec. (n=15; l\/l1SD)(n=15; M1sD)(n=15; M1sD)ULD anti-S100 + anti- 104.10113.14 ** 9.9011.20 * 96.90113.54 ***eNOSULD anti-S100 68.5016.57 >< 13.5011.51 194.20118.45 ><><> test 1 Visit 2 (course int ake) ULD anti-S100 + anti- 134,70120,24 ** 9,5011,38 ** 117,90115,65 **eNOS ULD anti-S100 82,70110,33 13,5011,69 167,50114,72 xULD anti-eNOS 74,3019,49 x 17,3012,40 209,20121,62 xxPlacebo 63,7013,91 15,0011,47 199,60131,19P value on Kruskal-Wallis 0,0341 0,0244 0,0061 test 1 Notes: 1 for determination of significant difference between groups the Kruskal-Wallis test was used. lf the test showed a significant difference of p <0.05 for comparison between groupsagainst each other the l\/lann-Whitney test was used.* the significant difference in comparison with placebo, p<0,05;** the significant difference in comparison with placebo, p<0,01;*** the significant difference in comparison with placebo, p<0,001.

X the significant difference in comparison with ULD anti-S1OO + anti-eNOS, p<0,05; xx the significant difference in comparison with ULD anti-S1OO + anti-eNOS,p<0,01; xxx the significant difference in comparison with ULD anti-S1OO + anti-eNOS,p<0,001.

The analysis of HRV at the rest condition (in sitting position) before and after theCCEAC test (Table 9) detected that in subjects receiving study drugs had a tendency toan increased rate of SDNN indicating an increase in heart rate variability due toparasympathetic influence on heart rhythm. ln response to a kinetic effect in alltreatment groups the value of Rl\/IS-SD increased which characterizes the activity of theparasympathetic component of autonomic regulation. ln the groups receiving thecomposition ULD anti-S1OO + anti-eNOS and ULD anti-S100 showed an increase in HFwhich also indicated a shift in autonomic balance toward parasympathetic link. Thus,after conducting CCEAC tests in all groups there was an increase of parasympathetic effects on heart rate.

Table 9The HRV parameters of the study participants at rest before and after the kinetic action 46 Visit 1 (one-day intake) Visit 2 (course intake) Parameter After the After the After the drug After thedrug intake CCEAC test intake CCEAC testULD anti-S100 + anti-eNOS group (IVI1SD)SDNN, msec. 57.715.51 68.217.42 59.415.03 65.614.66R|\/ISSD, msec. 43.116.77 51.419.22 47.016.21 47.615.33TP, msec. Z 979.01186.06 1678.31397.1 1067.21167.24 1381.01166.301#LF, msec. ¿ 437.51709.6 709.61178.72 391.9175.61 588.5187.48HF, msec 'Å 171.5151.08 228.4176.79 206.5158.32 218514396LF/HF, c.u. 4.210.82 4.910.83 3.310.83 4.210.91ULD anti-S100 group (IVI1SD)SDNN, msec. 60.914.62 70.915.90 59.114.80 68.814.87R|\/ISSD, msec. 44.315.39 50.616.56 42.414.63 47.815.57TP, msec. ¿ 832.21124.93* 1342.81217.0 841.41149.93 1288.01163.529 #LF, msec. Å 315.2152.38* 550.9172.44# 313616671 540.7187.57#HF, msec. ¿ 151.4141.19 247.0169.53# 138.3138.42 187.1139.80LF/HF, c.u. 3.010.54 4.010.72 2.810.53 4.010.52ULD anti-eNOS group (IVI1SD) SDNN, msec. 67.417.73 78.616.14 65.818.68 69.015.23 47 5 R|\/ISSD, msec. 53.018.86 58.417.68 59.6112.45 52.215.3OTP, msec. Å 1307.81324.24 1841 .11359.7 1232.31292.51 1275.41172.479#LF, msec. Å 576.51167.07 849.91194.2# 527.21167.07 562.1189.38HF, msec. Å 313.31139.9O 285.3165.92 218.9174.78 216.3163.72LF/HF, c.u. 3.610.87 3.910.82 3.711.14 3.810.58Placebo group (l\/|1SD)SDNN, msec. 64.616.1O 75.716.42 61.116.72 70.816.79R|\/ISSD, msec. 50.917.74 53.116.62 44.616.63 44.315.31TP, msec. Å 1062.21150.02 1917.81318.9 898.81169.62 1418.51227.596# # LF, msec. Å 440.6177.3O 832.41181.15 334817594 611.41113.64#HF, msec. Å 253.9159.95 266.7161.94 166.0148.14 174.1144.96LF/HF, c.u. 3.410.72 5.011.33 3.410.93 4.810.83 Note: *the significant difference in comparison with the placebo, pS0,05); # the significant difference in comparison with baseline parameters, ps0,05 The analysis of HRV in transition states showed that one-day intake ofcomposition ULD anti-S100 + anti-eNOS increased the reaction time (13.9 1 1.14; p s0.05) and the stabilization time (24.2 1 1.28; p S 0.05) in comparison with the ULD anti-S1OO and placebo. The same factors exceeded the value of the placebo group and afterthe kinetic effect which demonstrated the positive effect of the combined drug on thereactivity of the ANS (increase of tolerance to changes in body position). The smallest 48 difference between the maximum and minimum heart rate in the breath test confirmed abetter balance of the two divisions of ANS after receiving a one-day composition ULDanti-S100 + anti-eNOS (25_1 1 2.66 beats / min, p s 0.05). By the end of week course oftherapy the stabilizing effect on the balance of ANS after the CCEAC test (withorthostatic and breath test) is also noticed in the group receiving the composition ULDanti-S100 + anti-eNOS (Tables 10 and 11).

Table 10The HRV parameters of participants of the study at orthostatic test before and after kinetic action Visit 1 (one-day intake) Visit 2 (course intake)Parameter After drug After CCEAC After drug After CCEACintake test intake testULD anti-S100 + anti-eNOS (l\/l1SD) Group Exercise reaction, 1_3010_06 1.4010.04 1_3010_06 1_4010_06c.u.

Reaction time, sec. 13.911 .14*>< 12.711 _24* 11_810_57 11.711 _09Stabilization time, 24.211 .28*>< 21 .911 _44* 20.610.74 22.411 .44*> ULD anti-S100 (l\/l1SD) Group Exercise reaction, 1.4010.04 1.3010.04 1.3010.04 1.3010.05c.u.

Reaction time, sec. 7_6011_05 10.611 _55 9.711 _21 10.011 _73Stabilization time, 15.111 _16* 18.311 _43 18.011 _18 18.011 _80sec.

ULD anti-eNOS (l\/l1SD) Group Exercise reaction, 1.3010.04 1.3010.04 1.50 1 0.12 1.3010.04c.u.

Reaction time, sec. 8_2010_94 9_1011 _12 9.2 1 0.77 8_310_70Stabilization time, 16.511 _02 17.111 _33 19.0 1 2.04 16_710_98 49 SGC.

Placebo group (I\/|1SD) Exercise reaction, 1_3010_04 1_3010_04 1.40 1 0.06 1_3010_06c.u.Reaction time, sec. 9.511 _28 8_110_90 10.4 1 1.58 8.811 _09Stabilization time, 18_310_94 16.811 _09 18.0 11.37 16.511 _11sec.

Note: * the significant difference in comparison with placebo, pS0_05);>< the significant difference in comparison with ULD anti-S100, ps0_05_Table 11 The HRV parameters of participants of the study at breath test before and after kinetic action Visit 1 (one-day intake) Visit 2 (course intake)Parameter After drug After CCEAC After drug After CCEACintake test intake testULD anti-S100 + anti-eNOS (I\/|1SD) Group Corellation max HR / min HR, 1.5 1 0.05* 1.5 1 0.06 1.5 1 0.05 1.5 1 0.05 c.u.

Difference max HR - min HR, 25.1 1 2.66* 26.5 1 2.77 26.5 1 2.37 24.9 1 2.24*beats/min.

ULD anti-S100 (I\/|1SD) Group Corellation max HR / min HR, 1_510_06 1_610_05 1_510_04 1_610_06 c.u.

Difference max HR - min HR, 27.7*_f2.68 27.2*_«2.4O 25.7*_«2.24 26.9*_f2.67 beats/min.ULD anti-eNOS (MiSD) Group Corellation max HR/ min HR, 1.5*_«0.05 1.5*_«0.04 1.5:0.06 1.6*_f0.05 c.u.

Difference max HR - min HR, 26.7*_f2.44 2621204 27.7*_f2.47 27.3*_f2.12beats/min.

Placebo group (IVHSD) Corellation max HR/ min HR, 1.6*_f0.07 1.6*_«0.06 1.5:0.05 1.6:0.05 c.u.

Difference max HR - min HR, 3121306 2821250 27.7*_f2.37 29.2*_f2.44beats/min.

Note: *the significant difference in comparison with placebo, ps0,05 The results of self-esteem of functional state (well-being, activity, mood) of thesubjects which was conducted by the participants of the study after the simulation ofmotion sickness (CCEAC tests) at the beginning and at the end of therapy showed that the subjects of all the groups have given 'average' points for each of the parameters (Table 12). satisfactory. The highest growth rates compared with data of the placebo group by the end of the 7"' day of intake (more than 10%) was observed in the group of composition of ULD anti-S1OO + anti-eNOS.

Table 12 Thus, on the background of drugs intake the CCEAC tolerance was The dynamics of parameters of self-esteem of functional condition (welI-being-activity-mood) of study participants Parameter Visit 1 (one-day intake) Visit 2 (course intake) ULD anti-S100 + anti-eNOS (IVI1SE) group Well-being 4.310.26 4.610.27Activity 4.210.20 4.210.22Mood 5.010.16 521013 ULD anti-S100 (IVI1SE) group Well-being 3.71021 4.310.22Activity 3.610.'|7 4.010.19Mood 4.510.16 4.910.19 ULD anti-eNOS (IVI1SE) Group Well-being 3.910.25 4.11026Activity 3.810.25 3.910.23Mood 4.410.19 4.610.'|9 Placebo group (IVI1SE) Well-being 4.010.24 4.010.24Activity 3.810.20 3.710.26Mood 4.310.20 4.710.24 The safety analysis included data from a|| the subjects who participated in thestudy. During the observation period a well tolerance of studied preparations wasnoticed. No adverse events associated with drug administration identified. A|| the subjects of studied groups completed treatment in the terms established by the study protocol; there was not persons early dropped out.

According to the results of physical examination including indicators of heart rate,systolic and diastolic blood pressure and according to the Harvard step test data thesubjects were not recorded as with any abnormalities during the study (Table 13). Allidentified changes were not beyond the normal range. ln this case, subjectively all subjects reported satisfactory well-being.

Table 13 The dynamics of physical parameters and exercise tolerance of study participantsbefore and after kinetic action Visit 1 (one-day intake) Visit 2 (course intake)Parameter After drug After CCEAC After drug After CCEACintake test intake testULD anti-S100 + anti-eNOS (l\/l1SE) GroupHR (beats/min) 74.613.36 68.413.67 74.113.1O 67.712.62Systolic blood 123.412.83 125.914.08 121 .812.65 128.314.25pressure(mmhg.)Diastolic blood 74.013.09 79.312.62 76.212.43 80.313.3Opressure(mmhg.)Step-test index - 53.612.6O - 52.312.09ULD anti-S100 (l\/l1SE) GroupHR (beats/min) 73.512.57 69.712.78 72.112.84 67.712.39Systolic blood 127.512.55 133.514.77 127.112.55 129.915.06pressure(mmhg.)Diastolic blood 75.512.65 82.613.31 74.912.41 82.313.19pressure(mmhg.) Step-test index - 50.611 _71 - 53.011 _63ULD anti-eNOS (l\/l1SE) GroupHR (beats/min) 76_512_59 67.311 _98 77_312_O2 70_113_23Systolic blood 127_313_14 131 _515_16 123_513_O6 129_314_13pressure(mmhg_)Diastolic blood 75_212_24 80_312_66 73_912_83 81 _O13_22pressure(mmhg_)Step-test index - 51 _812_12 - 51 _212_21Placebo group (l\/l1SE)HR (beats/min) 74_512_78 68_913_46 73_913_23 72_313_58Systolic blood 125_313_30 133_314_73 124_312_83 12691395pressure(mmhg_)Diastolic blood 76_212_15 81 _712_83 75.411 _86 79_713_03pressure(mmhg_)Step-test index - 50_012_03 - 50.111 _99 ln addition to the hemodynamic parameters, for evaluation of the safety of studieddrugs and its possible negative impact on the central nervous functions, the followingphysiological parameters were examined in subjects: (Rl\/IO (reaction on moving object),Sl\/IRT (simple motor reaction time), RA (range of attention), attention span (AS), andattention stability factor (ASF))_ ln addition, the Stange's test was conducted to assess tolerance to hypoxia.

According to received results (Table 9) neither one-day or course drug intake hada significant effect on the estimated parameters. lndexes of sensory motor coordination(Sl\/IRT, Rl\/IO) did not differ from the results of the placebo group before and after the CCEAC test at both visits. Study data of such complicated functions like volume andstability of attention showed that the studied drugs both before and after the CCEAC testdid not change the degree of concentration and shift in attention not being different from the placebo group.

The analysis of standard exercise tests with breath holding showed a tendency toWhen holding the breath the duration of Stange's test grew after taking all study drugs. increase of the tolerance of hypoxia by the subjects (Table 14).However, onlyintake of the combination composition ULD anti-S100 + anti-eNOS showed significantlylonger time in the holding of the breath after the kinetic effect (68.1 1 18.8 sec. atbaseline and 91.7 1 27.4 sec. after the CCEAC test; p <0.05). The increase of toleranceof hypoxia was also noted when the Gench's test (Stange's test) (breath holding atexpiration, P> 0.05) was used.

Table 14 The dynamics of parameters of psycho-physiological state of study participants before and after kinetic action Visit 1 (one-day intake) Visit 2 (course intake)Parameter After drug After CCEAC After drug Afterintake test intake CCEACtestULD anti-S100 + anti-eNOS (M1SE) GroupSMRT 257.518.67 268.9110.18 269.619.75 279.9112.24RMO, c.u. 50.113.92 49.514.50 47.314.86 47.013.54rlïltvlO, % of target 3.010.95 4.511.15 5.311.58 4.011.11I AS, sec. 5.210.34 5.210.35 5.210.41 5.110.40Range of 41 .712.36 39.912.38 38.112.17 37.512.04attention, sec.ASF 17.411.66 17.211.51 18.011.71 18.811.72Stange's test 68.114.85 91 .717.07* 71 .816.02 85.519.36 Gench's test 47.114.03 50.113.94 46.713.28 48.114.52ULD anti-s1oo (MÉSE) Group SMRT 258.919.95 282.4113.56 268.411 1 _37 279.119.20RMO, c.u. 58.116.40 57.516.34 55.115.06 53.815.02älg/IQ, % of target 3.711.50 2.010.82 2.310.83 5.011.69 | AS, sec. 6.010.40 6.410.52 6.210.42 6.010.41Range of 42.612.68 42.112.27 42.712.30 41 .912.52attention, sec.

ASF 14.511.16 14.911.26 15.311.13 15.411.18Stange's test 59.014.09 7261619 64.514.93 75.915.67Gench's test 47.114.48 49.414.69 48.314.30 48.814.14ULD anti-eNOS (M1SE) group SMRT 257.718.49 279.4114.23 266.7113.19 275.5111.44RMO, c.u. 48.313.67 51.914.39 52.514.79 49.614.22älg/IQ, % of target 2.310.83 2.010.82 3.311.26 5.711.68 | AS, sec. 5.910.25 6.010.34 551024 5.910.33Range of 41 .912.10 43.812.39 41 .312.00 4251222attention, sec.

ASF 13.711.34 14.811.31 15.611.24 14.111.40Stange's test 62.515.49 69.515.09 56.713.34 73.117.98Gench's test 43.113.51 45.713.15 43.413.77 45.814.03Placebo group (M1SE) SMRT 267.617.64 290.1111.33 281.119.78 263.316.85RMO, c.u. 60.718.31 54.115.57 51.113.69 52.615.38älg/IQ, % of target 3.711.03 3.711.24 3.310.93 4.311.61 |AS, sec. 6.110.71 5.710.36 5.510.32 5.910.71 Range of 41 912.09 42.4i2.81 41 312.18 3961226attention, sec.

ASF 14.5i1.64 14.5i1.79 15.3i1.55 15.9i1.58Stange's test 63.7i4.71 67.9i6.90 64.8i5.94 83011224Gench's test 44.7i2.52 47.1i3.30 43.7i2.71 47.8i3.78 Thus, the study using an experimental motion sickness demonstrated theeffectiveness of the combination composition ULD anti-S1OO + anti-eNOS andmonocomponent preparation ULD-S100. The studied drugs increase the stability of thesubjects to the kinetic effect after simulation of the clinical and physiological effects ofmotion sickness contributing to more mild clinical process of motion sickness and earlierrecovery of the subjects after cessation of treatment. ln addition, it was shown that theanti motion sickness effect of the combination composition (compositions ULD anti-S1OO+ anti-eNOS) increases the efficiency of individual components. The effectiveness ofthe combination composition ULD anti-S1OO + anti-eNOS in the control of the vestibular-autonomic and sensory reactions of a body in experimental motion sickness increases atcourse intake. lt should be noted that ULD anti-eNOS in the form of monopreparationdoes not have a protective effect against motion sickness but when combined with ULDanti-S1OO significantly enhances the anti motion sickness effect of the last one whichmanifests itself as at one-day so at short course intake of the drug. The best ability toadjust the transient processes that is to influence to the reactivity of the parasympatheticand sympathetic parts of ANS as well as adaptive capabilities of ANS in a state ofmotion sickness (to increase the tolerance to sudden changes in a body position) wasobserved in the composition ULD anti-S1OO + anti-eNOS which is an importantcomponent of anti motion sickness properties of the drug. Composition ULD anti-S1OO +anti-eNOS and monocomponent preparation ULD anti-S100 when using them as antimotion sickness preparation including when performing an operator functions are safe and do not adversely impact on the physical and psycho- physiological parameters.

Combination composition ULD anti-S100 + anti-eNOS and ULD anti-S100 can berecommended for the prophylaxis and relief of kinesia in motion disease (including sea,air and car sicknesses) to persons with low and moderate degree of stability. Thecombination composition has high safety and no adverse effects on the quality of professional activity.

Example 9.

To study the properties of the combination pharmaceutical composition of thepresent application for the treatment of psychoorganic syndrome, tablets with weight of300 mg were used. The tablets were impregnated with pharmaceutical compositioncontaining water-alcohol solutions (6 mg/tab.) of activated-potentiated forms ofpolyclonal affinity purified rabbit brain-specific proteins antibodies S-100 (anti-S100) andto endothelial NO-synthase (anti-eNOS) in ultra low doses (ULD) obtained by superdilution of initial solution (witn oonoontration of 2.5 ing/ml) in 10012, 10000, 100200 times,of equivalent mixture of centesimal homeopathic dilutions C12, C30, C200 (“ULD anti- S100 + anti-eNOS”).

The control group patients received 300 mg tablets impregnated with(6 mg/tablet) ofactivated-potentiated forms of polyclonal affinity purified rabbit brain-specific proteinsantibodies S-100 (anti-S100) in ultra low doses (ULD) obtained by super dilution of initial solution (witn oonoontration of 2.5 ing/ml) in 10012, 10000, 100200 times. pharmaceutical composition containing water-alcohol solutions The study included patients diagnosed with psychoorganic syndrome ofposttraumatic origin. Psychoorganic syndrome is characterized by the following triad ofsigns: weakness of memory, loop of intelligence, incontinence of affect (Walther Bueltriad).

The study was an open-label randomized comparative parallel group clinical trialof efficacy and safety of the therapy in patients with psychoorganic syndrome ofposttraumatic origin (the first group of patients took the preparation of ULD anti-S100, the second group of patients -the preparation of ULD anti-S100+anti-eNOS).

The study included 6 patients aged 35 to 90 years old (mean age 70.83 i 21.95) diagnosed with psychoorganic syndrome.

Compliance of patients to following inclusion and exclusion criteria was checked: lnclusion criteria: Patients diagnosed with posttraumatic encephalopathy with psychoorganicsyndrome or with encephalopathy of complex etiology (vascular, posttraumatic)with psychoorganic syndrome, confirmed by medical history, neurologicalexaminations and medical records.

Patient without change in concomitant therapy within at least one month prior toVisit 1.

No need for change in concomitant therapy for the whole observation period.

No need for immunomodulatory drugs prescription for the next 6 months.

Patients with a level of education sufficient to adequately communicate with theresearcher and study coordinator.

Patients assessed by the researcher as reliable and ready to perform allscheduled clinical visits, tests and procedures stipulated in the protocol.

Patients having a valid home address.

Exclusion criteria: PWNF* Any brain surgery in medical history.

Acute myocardial infarction.

Hemorrhagic stroke.

The diagnosis of psychosis, bipolar disorder or schizoaffective disorder in medicalhistory. l\/lajor depressive disorder according to criteria of depression module ofinternational neuropsychiatric mini-interview (l\/l|Nl).

Factors/conditions of medical or another character which in the opinion of theresearcher may affect to the test results for patients in the study. 10.11.12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22.

Answers "2A", "2B", "2C" or "3" in the section "l" of Beck Depressionquestionnaire (active suicidal ideation with some intent to act, without a specificplan, or active suicidal ideation with a specific plan and intent).

Autoimmune disease in medical history.

Acute damage of liver or severe cirrhosis (class C by Child-Pugh).

Non-corrected disorder of thyroid gland function.

Decompensated arterial hypertension in medical history.

Serious or decompensated cardiovascular disease, liver disease, kidney disease,metabolic, respiratory or hematological disease, symptomatic peripheral vasculardisease or another medical or psychiatric condition which in the opinion of theresearcher, may affect the patient's participation in the study or could lead toprolonged hospitalization or re-hospitalization during the study.

Diseases and conditions which in the opinion of researcher may prevent patientfrom the participation in the study.

The intake of the drug containing ULD anti-eNOS or the drug containig ULD anti-S100 before inclusion in the study.

The intake of antidepressants of any group including plant and homeopathicpreparations.

The intake of anxiolytics of any group including plant and homeopathicpreparations.

The intake of immunomodulators including plant and homeopathic preparations.The treatment with systemic steroids within 1 month before Visit O.

The participation in the study of the drug containing ULD anti-eNOS or the drugcontaining ULD anti-S100 if patients took at least one doze of preparation.Participation in other clinical studies within 1 month before within 1 month beforebeing enrolled in this study.

Pregnancy, breast feeding, impossibility to use an adequate contraception duringthe study period and within 1 month after the last intake of the studied drug.

The presence of allergy/intolerance of any component of drugs including lactose intolerance. 23. Patients psychiatric diseases in patients. taking narcotic drugs and neuroleptics, alcoholic dependence, 24. Patients are the staff of the center which directly related to the conducted studyand/or are family members of the research center staff's which directly associatedwith the ongoing study. The "family members" are a husband (wife), parents,children, brothers (sisters). 25. Participation in the trial or presumable receiving of compensation or participation in the judicial process in the opinion of a researcher.

After the determination of patient conformity to inclusion and exclusion criteria thepatients were randomized into two study groups: a group of patients receiving ULD anti-S100 (3 patients, women - 33.33%, men - 66.66%, mean age - 71.33 i 16.25 yearsold), a group of patients receiving ULD anti-S100 + anti-eNOS (3 patients, women -66.66 % men - 33.33 %, mean age - 70.33 i 30.66 years old).

During this study the five visits were carried out. Treatment phase lasted fromVisit 1 to Visit 4 for 84 i 5 days on average. Visit 4 (Day 84 i 5) was the first endpoint ofthe study followed by a follow-up observation. Follow-up phase continued from Visit 4 toVisit 5 (Day 168 i 5 on average). ln the safety analysis the data of all patients participating in the study (n = 6) wasincluded. During the study good tolerance of the drug was recorded. No adverse eventswere registered. All patients of studied groups have completed the treatment accordingto the protocol; no early dropouts.

The effect of ULD anti-S100 + anti-eNOS preparation on the main clinical signsand symptoms of psychoorganic syndrome (NPl neuropsychiatric inventory, lntensitysection), on the intensity of concomitant distress of the person attending to the patient(NPl Neuropsychiatric lnventory, Distress section) as well as the on patient's cognitivefunctions (The Mini Mental State Examination, l\/ll\/ISE) were assessed. An improvement was found in the key symptoms of psychoorganic syndrome such as statistically 61 significant reduction of the intensity section of NPI neuropsychiatric inventory (from91.0*_f15.13 to 69.0*_f+6.24, p<0.05), decrease of distress section score of NPIneuropsychiatric inventory (from 44.33*_f17.78 to 36.33*_f3.21, p<0.05) at Visit 4 (Table15). ln the group of patients receiving ULD anti-S100 alone no clinical improvement was recorded.

At that, a difference between the groups of patients in the total score of theintensity section of NPI neuropsychiatric inventory at the end of therapy was statisticallysignificant at p<0.05.

Table15.NPI (intensity) NPl(distress) ADS-ADL l\/ll\/ISE ULD anti- S100+anti-eNOS before treatment 91 .O+15.13 44.33+17.78 42.66+4.93 22.33+3.21ULD anti- S100+anti-eNOS aftertreatment 69.0+6.244*# 36.33+3.21* 52.0+5.57 22.66+2.08ULD anti-S100 before treatment 114.0+25.53 45.66+14.47 33.0+13.89 22.33+4.16ULD anti-S100 after treatment 99.66+18.0 49.0+17.05 31 .66+10.69 23.0+4.36 * - p from baseline <0.05; # - p from control <0.05 Thus, in the conducted clinical study a positive effect of combined pharmaceutical composition ULD anti-S100 + anti-eNOS on the main clinical signs and symptoms of62 psychoorganic syndrome and tendency to effect cognitive functions with psychoorganicsyndrome. ln addition, good drug tolerability was confirmed. No drug-related adverseevents were registered.

Example 10.

To study the properties of the combination pharmaceutical composition of thepresent application for the treatment of Alzheimer's disease, tablets with weight of 300mg were used. The tablets were impregnated with pharmaceutical compositioncontaining water-alcohol solutions (6 mg/tabiet.) of activated-potentiated forms ofpo|yc|ona| affinity purified rabbit brain-specific proteins antibodies S-100 (anti-S100) andto endothe|ia| NO-synthase (anti-eNOS) in ultra low doses (ULD) obtained by superdilution of initial solution (with concentration of 2.5 mg/mi) in 10012, 10030, 1002°° times,of equivalent mixture of centesimal homeopathic dilutions C12, C30, C200 (ratio: 1:1)(“ULD anti-S100 + anti-eNOS”).

The control group patients received 300 mg tablets impregnated with(3 mg/tablet) ofactivated-potentiated forms of po|yc|ona| affinity purified rabbit brain-specific proteinsantibodies S-100 (anti-S100) in ultra low doses (ULD) obtained by super dilution of initial solution (with concentration of 2.5 mg/ml) in 10012, 10030, 100200 times, of equivalent pharmaceutical composition containing water-alcohol solutions mixture of centesimal homeopathic dilutions C12, C30, C200 The study included patients diagnosed with Alzheimer's disease. Alzheimer'sdisease is characterized by dementia (acquired dementia, stable impairment of cognitiveactivity with certain loss of previously acquired knowledge and practical skills, difficulties or impossibility to gain new knowledge).

The study was an open-label randomized comparative clinical trial of efficiencyand safety of the therapy in two parallel groups (preparations of ULD anti-S100 and ULDanti-S100+anti-eNOS) in the treatment of patients with mild to moderate Alzheimer'sdisease. 63 The study included 6 patients aged 55 - 64 years old (mean age 59.0 i 3.58) diagnosed with mild to moderate Alzheimer's disease.

Compliance of patients to following inclusion and exclusion criteria was checked: lnclusion criteria are as follows: Patients with mild to moderate Alzheimer's disease, confirmed by medical history,neurological examinations and medical records.

No need for change in concomitant therapy for the whole observation period.

No need for immunomodulatory drugs prescription for the next 6 months.

Patients having a valid home address.

Exclusion criteria are as follows: PWNF* Any brain surgery in medical history.

Acute myocardial infarction.

Hemorrhagic stroke.

Factors/conditions of medical or another character which in the opinion of the researcher may affect to the test results for patients in the study. 64 10.11.12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22.

Autoimmune disease in medical history.

Acute damage of liver or severe cirrhosis (class C by Child-Pugh).

Non-corrected disorder of thyroid gland function.

Decompensated arterial hypertension in medical history.

The intake of the drug containing ULD anti-eNOS or the drug containig ULD anti-S1OO before inclusion in the study.

The presence of allergy/intolerance of any component of drugs including lactose intolerance. 23. Patients psychiatric diseases in patients. taking narcotic drugs and neuroleptics, alcoholic dependence, 24. Patients are the staff of the center which directly related to the conducted studyand/or are family members of the research center staffs which directly associatedwith the ongoing study. The "family members" are a husband (wife), parents,children, brothers (sisters). 25. Participation in the trial or presumable receiving of compensation or participation in the judicial process in the opinion of a researcher.

After the determination of patient conformity to inclusion and exclusion criteria thepatients were randomized into two study groups: a group of patients receiving ULD anti-S100 (3 patients, women - 100%, men - 0%, mean age - 59.0 i 3.6 years old), a groupof patients receiving ULD anti-S100 + anti-eNOS (3 patients, women - 66.66 % men -33.33 %, mean age - 59.0 i 4.36 years old).

The effect of ULD anti-S100 + anti-eNOS preparation on the main clinical signsand symptoms of Alzheimer's disease (NPl neuropsychiatric inventory, lntensitysection), on the intensity of concomitant distress of the person attending to the patient(NPl Neuropsychiatric lnventory, Distress section) as well as the on patient's cognitivefunctions (The Mini Mental State Examination, l\/ll\/ISE) were assessed. An improvement was found in the key symptoms of Alzheimer's disease such as statistically significant 66 reduction of the intensity section of NPI neuropsychiatric inventory (from 24.33: 4.73 to12.0*_f3.46, p<0.05) at Visit 4 (Table 16).

A tendency for reduction of distress of the person attending to the patient wasalso found as well as for the reduction in activity of the patients everyday life at the endof therapy (however, without any statistically significant difference, possibly due to thesmall number of patients included in the study).

Besides, a tendency for improvement of cognitive functions was found,manifested by increase of l\/ll\/ISE score from 23.66*_f3.21 to 266611 .53 points, however,the difference also failed to reach statistically significant values at the end of therapy, which may also be related to the small sample size.

The same endpoints in the group of patients receiving ULD anti-S100, showed notrend for improvement, except a statistically insignificant improvement of MMSE scorefrom 22.66iO.58 to 23.33*_fO.58 points.

At that, a difference between the groups of patients in the total MMSE score atthe end of therapy was statistically significant at p<0.05.

Table16.NPl (intensity) NPl ADCS-ADL l\/ll\/ISE(distress) ULD anti- 24.33*_f4.73 9.66*_f1.53 71.0*_f6.56 23.66*_f3.21S100+anti-eNOS before treatment ULD anti- 12.0*_f3.46* 5.0*_f3.61 74.33*_f2.51 26.66*_f1.53#S100+anti-eNOS after treatment ULD anti-S1OO 35.66*_f5.5O 22.33*_f5.5O 61.66*_f5.13 22.66*_fO.58before treatment ULD anti-S1OO 38.33*_f8.5 23.0*_f5.0 61.33*_f5.86 23.33*_fO.58after treatment * - p from baseline <0.05; # - p from control <0.05 67 Thus, in the conducted clinical study a positive effect of combined pharmaceuticalcomposition ULD anti-S100 + anti-eNOS on the main clinical signs and symptoms ofAlzheimer's disease and tendency to effect cognitive functions with Alzheimer's disease.ln addition, good drug tolerability was confirmed. No drug-related adverse events wereregistered.

Example 11.

Group 1 - the active drug group was given 300 mg tablets impregnated with anaqueous-alcohol solutions (6 mg/tab) of activated-potentiated form of polyclonal rabbitantibodies to brain specific S-100 protein (anti-S-100), and to endothelial NO-synthase(anti-eNOS) in ultra low dose (ULD anti-S-100 + ULD anti-eNOS), purified on antigen,obtained by super dilution of initial solution (with concentration of 2.5 mg/ml) in 10012,10030, 100200 time, equivalent to mixture of centesimal homeopathic dilutions C12, C30,C200; Group 2 - the comparison group was given 300 mg tablets impregnated with anaqueous-alcohol solution (3 mg/tab) of activated-potentiated forms of polyclonal rabbitantibodies to brain-specific S-100 protein purified on antigen in ultra low dose (ULD anti-s100) ooteineo by super oiiution of initiei soiution in 10012, 10030, 10050 times, ofequivalent mixture homeopathic dilutions C12, C30, C50.

Group 3 - the control group (placebo) was given of 300 mg tablets havingexcipients (lactose monohydrate - 267 mg, microcrystal cellulose - 30 mg, magnesiumstearate - 3 mg).

The effectiveness of the active drug ULD anti-S100 + anti-eNOS in the treatmentof patients with syndrome of attention deficit and hyperactivity disorder (ADHD) wasconducted in comparative double blind placebo-controlled study in 146 children from 6 to12 years old (mean age 9.3*_f0.24 years old) who were randomized into three groupsdepending on prescribed therapy. Within 12 weeks the patients of group No.1 (n = 46) 68 received the composition ULD anti-S100 + anti-eNOS, 2 tablets twice a day; thecomparison group 2 members (n = 50) received ULD anti-S100, 2 tablets twice a day;the control group 3 members (n = 50) received 2 tablets twice a day. All the patientsincluded in the study had clinically marked presentations of ADHD which was confirmedby high points on ADHD symptoms assessing scale (ADHDRS-IV-Home Version): 33.8i 0.92 in group 1; 32.5 i 1.14 in group 2 and 33.6 i 0.91 in group 3. Most of thechildren were characterized by a moderate degree of severity of ADHD according to theCGI-ADHD-Severity questionnaire. The total score on this scale was 4.0 i 0.02 points inthe group 1, 4.0 i 0.03 points in the group 2, and 4.0 i 0.00 points in the group 3. Thus,initially the patients of the three groups had comparable indicators of the severity ofADHD. According to the results of neurological, clinical - laboratory and instrumentalexamination at the time of enrollment to the study no abnormalities in any patient wasdetected. Over the 12 weeks of treatment, patients were seen six times by a doctor.During which the physician-researcher recorded the dynamics of intensity of clinicalpresentations of ADHD (total score on a scale ADHDRS-IV-Home Version) and diseaseseverity (on the CGI-ADHD-Severity), supervised the prescriptions and administration oftreatment and evaluated the safety of the treatment.

The analysis of the effectiveness of 12 weeks of therapy in the three groupsshowed a decrease of more than 25% from the initial total score on a scale ADHDRS-lV-Home Version in 75% (n = 36) of children treated with the composition ULD anti-S100 +anti-eNOS; in 66% (n = 33) of patients treated with ULD anti-S100 and in 56% (n = 28)of children receiving placebo. Differences of efficiency between the groups showing amore detailed assessment, taking into account the three-level grading of improvement ofcondition (reduction of total score on a scale ADHDRS-IV for <25%, 25-49.9% or 2 50%from the baseline), are presented in Table 17. Significant improvement with a reductionin total score on 50% or more from the baseline was noted in 52% of children in group 9who were taking ULD anti-S100 + anti-eNOS, and in 34% of children in group 2 whowere taking ULD anti-S100 (vs. 8% of patients in group 3 with placebo).

Significant reduction (p <0.001) of clinical implications of ADHD in comparison with the initial state is already occurred after 2 weeks of therapy in all three groups of69 observation. Positive dynamics was more significant in patients of groups 9 and 2 as thesignificant differences were identified in them between total scores ADHDRS-IV-HomeVersion, not only in relation to the screening visit but when compared with the indexes ofthe group 3 with placebo. ln subsequent weeks of treatment the efficacy of treatmentwith composition ULD anti-S100 + anti-eNOS and monocomponent preparation ULD-The resulting decrease in total score on a scale ADHDRS-IV-Home Version in children of the S100 started to grow, the most significantly in the active drug group (p <0.05). group 9 with ULD anti-S100 + anti-eNOS was 16.5 points, in patients of the group 2 withULD anti-S100 - 12.4 points (compared to 6.3 points in the group 3 with placebo). As aresult of 12-week of treatment the intensity of clinical implications of ADHD in childrentreated with the composition ULD anti-S100 + anti-eNOS decreased by almost in half (-48.8%) and in patients treated with ULD anti-S100 more than in one-third (-38.2%)compared with the baseline.

The intake of composition ULD anti-S100 + anti-eNOS or ULD anti-S100influenced on both clusters of symptoms of ADHD which was confirmed by dynamics ofassessments by two sections of the scale with ADHDRS-IV-Home Version. l\/loreover,the treatment with the composition ULD anti-S100 + anti-eNOS was significantly higherthan the effectiveness of therapy with monopreparation ULD anti-S100 in the degree ofinfluence on the and attention deficit and intensity of implications hyperactivity/impulsivity.

The positive therapeutic effect of the active drug ULD anti-S100 + anti-eNOS anddrug of comparison ULD-S100 was shown in evaluating of patients' treatment results ona scale of ADHD severity assessment (CGI-ADHR-Severity) (Table 17). Almost thefourth part of the patients in ULD anti-S100 + anti-eNOS group the severity of diseasewas decreased from moderate to mild and even to minimal as confirmed by a decreasein mean value on a scale CGI-ADHR-Severity on 15% after 3 months of therapy (from4.0 i 0.02 to 3.4 i 0.06; p <0.001). The effect of therapy with monopreparation ULDanti-S100 was slightly lower and indicated -10% on a scale CGI-ADHR-Severity over 3months (vs. 5% in the placebo group). The safety analysis included data of all the patients participating in the study. During the whole period of monitoring there was both, well comparable to placebo, the tolerance of active drug ULD anti-S1OO + anti-eNOSand preparation of comparison ULD-S100. Adverse events were reported in one patientof the group with ULD anti-S1OO (subside during the fourth week of the studyheadaches) and in one patient of the placebo group (sieepwaiking during the secondmonth of observation). These adverse events were not connected with the therapy. lnaddition, during the treatment the single cases of acute respiratory disease wereobserved which also are not associated with the therapy. All the patients of studiedgroups completed the treatment to schedule established by the study protocol; no earlydropouts. The absence of pathological changes according to physical examination ofthe patients and in the course of repeated analysis of laboratory parameters confirmedthe safety of studied therapy.

According to the results of physical examination (heart rate, SBP, DBP, bodytemperature) in patients any pathological alterations during treatment were notregistered. Differences in analyzing rates according to visits and in the comparedgroups did not reach the statistical significance and do not exceed the limits ofphysiologically-allowable deviations. High rates of adherence to therapy additionallyevidenced as about effectiveness so as about the safety of studied preparations. By theend of the third month of treatment the adherence was 99.8 i 1.15% and 98.8 i 2.25%in the group 9 with ULD anti-S1OO + anti-eNOS and in the group 2 with ULD anti-S100 respectively (versus 74.6 i 2.54% in the group 3 with placebo).

Thus, the study demonstrated the efficacy and safety of the compositions ULDanti-S1OO + anti-eNOS and of monocomponent preparation ULD-S1OO in the treatmentof children with ADHD. The most pronounced therapeutic effect in the 12-week coursewas observed in complex drug (ULD anti-S1OO + anti-eNOS) which was manifested byThecomposition ULD anti-S1OO + anti-eNOS had correcting influence to both of the clusters positive dynamics of clinical symptoms in the majority (75%) of children. of symptoms of ADHD and as a result, the significant reduction of attention disordersand hyperactivity in patients with ADHD was noted. 71 Table 17. The dynamics of total score by the scale ADHDRS-IV-Home Version by the end of 12 weeks of therapy The proportion of patients with decrease of total score by the scale ADHDRS-IV-Home Version The difference is significant in comparison with the placebo group: s i” p Gmups °f patients Less than on 25.0 - on 50.0% and25.0% from 49.9% from more frombaseline baseline baseline ULD anti-S100 + anti- ##12 (25%) 11 (23%) 25 (52%) eNOS, n=48 ULD anti - S100, n=50 17 (34%) 16 (32%) 17 (34%) m* Placebo, n=50 22 (44%) 24 (48%) 4 (8%) Table 18. The dynamics of evidence of clinical implications of ADHD by the scaleADHDRS-IV-Home Version 72 ULD anti-S100 + ._ ULD t -S100, =50 Pl , = Treatment antheNos, n=48 an | n acebo n 50stage Value A from Value A from Value A from (MiSE) baseline (MiSE) baseline (MtSE) baseline Total score 33.8 32.5 33.6Screening 10.96 i 1.14 i 0.91 24.1 -28.7% 251 2882 Weeks -rf-k* 0/ 0/ š°97 i 1.03 o i 1.26 o *** # *** 22.6 -33.1% 22.7 29.94 Weeks :#098 *** i 1.23 -30.2 % i 1.06 -11.0 %*** ## ***19.4 -42.6% 20.8 29.06 Weeks :#095 *** i 1.06 -36.0 % i 1.25 -13.7 %*** ## ***18.9 -44.1% 20.9 27.68 Weeks šå94 *** i 1.30 -35.7 % i 1.35 -17.9 %*** ### ***17.3 -48.8% 20.1 27.312 Weeks ä0å6 *** i 1.21 -38.2 % i 1.48 -18.8 %*** ### ***Attention disorders18.4 17.4 18.4Screeningi0.55 i 0.57 i 0.4312.8 -30.4% 13.7 16.12 Weeks i0.š7 i 0.68 -21.3 % i 0.66 -12.5 %*** # ***11.6 -37.0% 12.9 16.44 Weeks iOÉg i 0.79 -25.9 % i 0.57 -10.9 %*** *** ### ***10.7 -41.8% 11.96 weeks i0.64 i 0.64 -316 % 160 f.. -130 %*** ### ###10.3 -44.0% 115 1518 weeks -33.9 °/ -17.9 °/fﬁšå i 0.70 o i 0.76 o 73 *** ### *** 9.7 -47.3% 11.4 14.9 12 Weeks i0.šš& i 0.68 -34.5 % i 0.78 -19.0 %*** *** ## *** Hyperactivity /impulsion 15.4 15.1 15.2 Screeningi0.61 i 0.77 i 0.6211.3 -26.6% 12.7 11.4 2 WeekS i0.63 -24.5 % i 0.74 -16.4 % i 0.61 11.0 -28.6% 9.8 13.5 4 WeekS i0.62 i 0.64 -35.1 % i 0.67 -11.2 %*** ### *** ### **8.7 -43.5% 8.9 12.9 6 WeekS i0.59 i 0.64 -41.1 % i 0.73 -15.1 %*** ## *** ### **8.6 -44.2% 9.5 12.5 8 WeekS i0.60 i 0.76 -37.1 % i 0.81 -17.8 %*** ## *** ## ***7.6 -50.6% 8.7 12.5 12 WeekS i0.57 i 0.70 -42.4 % i 0.82 -17.8 %*** ###& *** ### *** Note. The difference is significant in comparison with baseline parameter: * p The difference is significant in comparison with placebo group: f* p 74 The difference is significant in comparison with the group of u|_D anti-emo; 8* p Table 19. The dynamics of severity level of ADHD by the scale CGI-ADHD-Severity ParameterADHD SeveritylVliSE A from baseline ULD anti-S100 + anti-eNOS, n=48 Screening 4.0*_f0.02 4 Weeks 3.6*_f0.02** -10%12 Weeks 3.4*_«0.06*** -15%ULD anti-S100, n=50 Screening 4.0*_f0.03 4 Weeks 3.8*_f0.06** -5% 12 Weeks 3.6*_f0.08*** -10%Placebo, n=50 Screening 4.0*_f0.01 4 Weeks 3.9*_f0.05 -2.5%12 Weeks 3.8*_f0.06*** -2.5% The difference is significant in comparison with the baseline parameter: ** p<0.01, ***p<0.001.

Example 12.

Double blind, placebo-controlled clinical study of a combination of activatedpotentiated forms of antibodies to the C-terminal fragment of the angiotensin ll AT1-receptor, in a mixture of homeopathic dilutions of C12, C30, C200, with activated potentiated form of antibodies to endothelial NO-synthase, in a mixture of homeopathic 76 dilutions of C12, C30, C200, in human patients with chronic heart failure to evaluate keyparameters of the CHF pathology. 80 patients (CHF of ||-|V functional class (FC), left ventricular ejection fraction(LVEF) less than 40%) were divided in 4 equal treatment and control groups for a 6months study. The background therapy was not discontinued (bisoprolol ß-blocker,ACE inhibitor enalapril, aspirin (unless contraindicated); administration of diuretics,nitrates, digoxin was also admitted). Group 1 received the activated potentiated form ofantibodies to a C-terminal fragment of the angiotensin ll AT1-receptor (mixture ofhomeopathic dilutions C12, C30, C200) (3 tablets/day, n=20).activated potentiated form of antibodies to endothelialhomeopathic dilutions C12, C30, C200) (3 tablets/day, n=20). Group 3 received the combination pharmaceutical composition comprising both activated potentiated form of Group 2 received theNO-synthase (mixture of antibodies to a C-terminal fragment of angiotensin ll AT1-receptor (mixture ofhomeopathic dilutions C12, C30, C200) and activated potentiated form of antibodies toendothelial NO-synthase (mixture of homeopathic dilutions C12, C30, \ C200) (3tablets/day, n=20). comparable in the initial study parameters: in age and sex, and severity (class of CHF Group 4 received placebo (3 tablets/day, n=20). The groups were and LVEF) and duration of the disease.

Before and after treatment, the patients were evaluated for the effect of theadministered medications on vascular remodeling and endothelium dysfunction that isimportant for the CHF process and progression. The effects of the medications on theprocesses of vascular remodeling were evaluated by pulse wave velocity (PWV)(“Colson” system) in the carotid-femoral (CF) (elastic type) and carotid-radial (CR)(muscle type) segments of arteries.

Table 20 shows the dynamics in the rates of pulse wave velocity in the carotid-femoral (CF) (elastic type) and carotid-radial (CR) (muscle type) segments of arteries.

Table 20Groups/ ULDs” of Abs¿ to ULD of Abs to Combination of PlaceboParameters C-terminal endothelial NO- ULDs of Abs tofragment of AT1 synthase C-end fragmentreceptor of of AT1 receptor 77 angiotensin ll of angiotensine ll andULDofAbs to endothelial NO-synthase^ & A% ^ & A% ^ & A% ^ A% CF,m/c 9, 8: -14.8* 10 9.8: - 10. 8.6 - 8. 8.2:0. 0.17: 0.6 .1 0.4 2.9 8: :0. 20.3* 2: 50. :0 7 0.3 6 0.5 .5 4CR,m/c 8. 8.9 2.9 8. 8.3: - 8.9 7.6 - 9. 9.7:0. 6.4* 6: :0. 8: 0.3 5.7 :0. :0. 15.6* 1: 30. 3 0. 5 7 #$ 0.2 1 3 (^) denotes initial value (&) denotes 6 month after beginning of administration (*) denotes difference from initial value is verifiable with p value < 0.05. (#) denotes difference from the group receiving ULDs of Abs to C-terminalfragment AT1 receptor angiotensin ll with verifiable difference in p value of < 0.05. ($) denotes difference from the group receiving ULDs of Abs to endothelial NO-synthase with verifiable difference in p value of < 0.05. (1) ULD denotes ultra-low doses. (2) Abs denotes antibodies.

After 6 months of treatment, only group 3 showed a proven effect of the claimedpharmaceutical composition on the stiffness of muscular type arteries. Group 1 whichreceived ULD of antibodies to a C-terminal fragment of angiotensin ll AT1-receptor, andgroup 3 which received the combination pharmaceutical composition of the invention showed a proven increase in the stiffness of elastic type arteries.

Example 13.

Double blind, placebo-controlled clinical study of a combination of activatedpotentiated forms of antibodies to the C-terminal fragment of angiotensin ll AT1-receptor, in a mixture of homeopathic dilutions of C12, C30, C200, with activatedpotentiated form of antibodies to endothelial NO-synthase, in a mixture of homeopathicdilutions of C12, C30, C200, in human patients with chronic heart failure to evaluate key measurement of quality of life.78 80 patients (CHF of ||-|V functional class (FC), left ventricular ejection fraction(LVEF) less than 40%) were divided in 4 equal treatment and control groups for a 6months study. The background therapy was not discontinued (bisoprolol ß-blocker, ACEinhibitor enalapril, aspirin (unless contraindicated); administration of diuretics, nitrates,digoxin was also admitted). Group 1 received the activated potentiated form ofantibodies to a C-terminal fragment of angiotensinhomeopathic dilutions C12, C30, C200) (3 tablets/day, n=20).activated potentiated form of antibodies to endothelialhomeopathic dilutions C12, C30, C200) (3 tablets/day, n=20). Group 3 received the combination pharmaceutical composition comprising both activated potentiated form of ll AT1-receptor (mixture ofGroup 2 received the NO-synthase (mixture of antibodies to a C-terminal fragment of angiotensin ll AT1-receptor (mixture ofhomeopathic dilutions C12, C30, C200) and activated potentiated form of antibodies toendothelial NO-synthase (mixture of homeopathic dilutions C12, C30, \ C200) (3tablets/day, n=20). comparable in the initial study parameters: in age and sex, and severity (class of CHF Group 4 received placebo (3 tablets/day, n=20). The groups were and LVEF) and duration of the disease. Before and after treatment, the patients wereevaluated for the life quality (l\/linnesota and Kansas questionnaires), morphologicalparameters of the heart, and tolerance to physical exercise.

Table 3 shows the results of the study in the form of dynamics in the basicparameters of the treatment efficacy.

After 6 months of treatment, the patients in group 1 treated with ULD ofantibodies to a C-terminal fragment of angiotensin ll AT1-receptor showed a significantimprovement of the life quality, improvement of the left ventricular systolic function, andan increased tolerance to physical exercise. Group 2 showed a proven decrease in theanxiety and depression levels and in the life quality, which were evaluated using theKansas questionnaire. The study confirmed that the maximum therapeutic effect wasachieved with the combination pharmaceutical composition of the invention incombination with the standard CHF therapy, which was administered to patients from group 3 that showed a proven positive dynamics in all parameters under study. 79 The combination of activated (potentiated) forms of antibodies to a C-terminal fragment of angiotensin ll AT1-receptor and to endothelial nitric oxide synthase (NO- synthase) in the pharmaceutical composition of the invention (combination drug) provides an unexpected synergistic therapeutic effect implying an enhanced influence on vascular remodeiing and endotheiium dysfunction that is critical for the CHF process and progression, as also on the improvement of the patients' life quality, on morphological parameters of the heart and tolerance to physical exercise, which is confirmed by clinical trials.The results are set forth in Table 21.

Table 21Groups/ ULD1ofAbs¿to C- ULD ofAbsto Combination ofULD of PlaceboParameters terminal fragment endothelial Abs to C-terminalof AT1 receptor of NO-synthase fragment of AT1angiotensin ll receptor ofangiotensin ll andULD ofAbs toendothelial NO-synthase_ ^ & A% ^ & A% ^ & A% ^ & A%l\/linnesota¿40.47.5 39.1 48 gi 43 32% 42.41 13.8 - ' - ' _ 48-3 12.9 _17.6 3.8 15.2 ä; 4-9 271 13.7 122 1* - - ***$ 1*Kansas472.82.1 70.1 65.717.81. 87,i *5-5 ' 5+2 02i8 ' 7+ 3 -251 838 603 -7214.6 5- ' 11.7 2-3 ' 13.5 16.8 ' 11* - * - ***$ 80 HADsb153 11.+' 12-5 _ 16- 34: _ 16. 6.4:0.9 173 159- :0.9 135 2:1 2.1 303 2: -46.1 +11 +11 -6.1** ' .7 ' 1.3 ***#$$ _ ' _' sm*FCCHF”27* ä? - 2.9: 2.7: _73 fo mio-l _3156 2.7: 2.5:0 _6201 H 17.3 0.1 0.2 02 4441113 0.1 .1FFLv'27-1 34.6i133. 25. 25.i 6:1 24.0 282 3:1 10.3 3: '9 36.7 26-4ï 280 6.35.... :1.5 7 1 1 1.1 :1.40.9 - - - ***#$6-minute walktest376. ål? 363. 416. 377 450.1: 390.5 40917:1 3-7* 10.6 1: 6:1 6.6 +1 17.7** 16.9 : +11'5 4.62.4 1,; 15.3 7.2 54 #6 11.9 _ ' *, **, *** - p Values < 0.05, 0.01 and 0.001, respectiVely #- difference from group receiVing ULDs of Abs to C-terrninal fragment ATl ofangiotensin receptor ll With Verifiable With p Value < 0.05 $, $$ - difference from the group receiVing ULDs of Abs to endothelial NO-synthase isVerifiable at p values of 0.05 and 0.01, respectiVely. (1)-ULD means ultra low doses (2) Abs means antibodies (3) “Minnesota” denotes Minnesota Questionnaire 81 (4) “Kansas” denotes Kansas Questionnaire (5) HADS denotes HADS total score (6) FC CHF denotes patients With chronic heart failure, functional class(7) FF LV denotes fraction of ﬁulnctioning of left Vertical.

Example 14.

To study properties of the proposed pharmaceutical composition in the treatmentof patients with a benign prostatic hyperplasia, 300 mg pills were used, saturated withthe pharmaceutical composition containing water-alcohol solutions (6 mg/pill) ofactivated - potentiated rabbit polyclonal affinity purified antibodies to prostate specificantigen (anti-PSA) and endothelial NO synthase (anti-eNOS) in ultra low doses (ULD),produced by ultra olilution of tne initial matrix solution 10012, 10000, 100200 times,equivalent to the mixture of centesimal homeopathic dilutions C12, C30, C200 (ULDanti-PSA +anti-eNOS), and 300 mg pills, saturated with the pharmaceutical compositioncontaining water-alcohol solutions (3 mg/pill) of activated potentiated rabbit polyclonalaffinity purified antibodies to prostate specific antigen in ultra low doses (ULD), obtainedby an ultra olilution of tne initial matrix solution 10012, 10000, 100200 times, equivalent tothe mixture of centesimal homeopathic dilutions C12, C30, C200 (ULD anti-PSA).

Benign prostatic hyperplasia (BPH) is one of the most frequently occurringdisorders in males (Bruskewitz R.C., 2003; Rosen R., 2003): on the one hand,epidemiological studies, carried out in Russia, point to a gradual increase in frequency ofBPH from 11.3% in 40-49 year olds to 81.4% in 80 year olds (Gorilovskiy, L.l\/l., 1999);on the other hand, demographic studies conducted by WHO confirm a significantincrease in the population over 60 years old, surpassing any other age group growth.

The main symptoms of benign prostatic hyperplasia are lower urinary tractsymptoms, which can cause significant discomfort and decrease quality of life(Bruskewitz R.C., 2003; Lepor H., 2004; O'Leary l\/l.P., 2005). ln severe cases, thedisease can be accompanied by complications, such as acute urinary retention, urinarytract infection, erythruria, kidney failure (Stepanov, V.N., 1999; Jacobsen S.J., 1997; 82 Lepor H., 2004). BPH is also associated with development of erectile dysfunction inpatients (Bruskewitz R.C., 2003; Daly l\/IP, 2005).

An open-label comparative parallel group study of efficacy and safety the ofpharmaceuticai compositions containing ULD anti-PSA + ULD anti-eNOS and ULD anti-PSA in ameiiorating urinary disturbances to caused by benign prostatic hyperpiasia(BPH), included 40 patients selected in accordance with inclusions/exclusions criteria.Patients were randomized in 2 groups, one group received 1 pill 3 times per day during12 weeks (n=21) of a ULD anti-PSA +anti-eNOS, and another one 1 pill 3 times per dayduring 12 weeks (n=19) of a ULD anti-PSA. The groups were comparable in age,severity of BPH symptoms, urination parameters and prostate volume.

The study included patients over 45 years old with a history of BPH withcorresponding symptoms of lower urinary tract for no less than 6 months, lPSS213,prostate volume according to transrectal ultrasonography 230 cm3, with maximumurinary flow speed of 24 ml/s and S15 ml/sc and minimum residual urine volume equal to125 ml, with PSA level S4 ng/ml. A necessary inclusion criterion was absence of intakeof the following medications in the medical records: finasteride, dutasteride, or otherexperimental drug 6 months prior to inclusion in the study, oi1-adrenoreceptor blockersand herbal medications 4 weeks prior to the inclusion into the study, any inhibitors ofphosphodiesterase type 5 and other erectile dysfunction treatments 4 weeks prior to theinclusion into the study.

The study did not include patients undergone invasive methods of treatment ofBPH, including transurethral prostatic resection, thermotherapy, transurethral needleablation, stent angioplasty and other; with malignant oncological disease, acute urinationdelay, bladder stones, urethral stricture, l\/larion's disease, genitourinary systeminfections in the phase of active inflammation and others.

Clinical efficacy of pharmaceuticai compositions was assessed by the improvementof clinical symptoms of lower urinary tract, evaluated using IPSS questionnaire(International Prostate Symptom Score), urination parameters (maximum and average urinary flow speed, urination volume, volume of residual urine) and prostate volume83 based on the data of transurethral ultrasound (TU), and also erectile function wasevaluated based on the data obtained from IIEF questionnaire (International Index ofErectiie Function). Results of the study are shown in tables 22 and 23. 84 Table 22.

ULD anti-PSA ULD anti-PSA + ULD anti-eNOSn/N(%) ln., 12 A, n/N ln., 12 A, cp1 aver. weeks. cp (%)1 aver. weeks.,aver. ,aver.'Psasfmre 19/19 17.3 11.9 -5.9 2805/221 13.0 10.5 -5.3(1000) l -)QoL/, score19/19 20/21(quamyof (1080) 3.4 2.4 -1.0 (852) 3.4 2.3 -1.1life)IIEF, score 2/19 4/21(185) 17.3 13.3 0.3 (198) 17.5 13.9 1.4Qmax,ml/s16/19 15/21(maximum (842) 10.8 13.1 2.2 (714) 11.7 13.7 2.0urine rate)Qave,ml/s15/19 18/21(average (788) 5.3 7.1 1.3 (857) 5.3 7.1 1.3urinerate)V, ml10/19 - 15/21 203.(Vcﬂumeof (528) 213.3 203.3 118 (714) 7 252.0 43.3unnaüon)RV,m|(reßidual 1549 23.3 19.4 -4.3 14/21 19.1 14.1 -5.0volume of (789) (656)urine)Pv,cm~*18/19 15/21(prostate (847) 55.9 43.9 -7.0 (714) 57.0 52.4 -4.3volume) 1- the numerator is a number of patients (n) showing improvement, denominator istotal number of patients in the study (N). 5 Table 23.Dynamics of subscales of obstructive and irritative symptoms, and question 7 ofIPSS questionnaire ULD anti-PSA ULD anti-PSA +anti-eNOSivitso visitivitso visit 1 ivitso visit 2 1 ivitso visit 2obstructive 1o.o»_«3.o2# 6.5t2.s1 s.2t2.96 6.ot3.39**irrit. 7.5t2.21& 5.3t1.9o*** 7.st2.16& 4.5t2.34***7” question 2.1 tors 1.9»_«o.75 2.3»_fo.9o 1.4»_«o.9s***obstr., %2 -33.4»_«26.s5 -25.2»_«34.5oirrit., %2 28.211 730 -4o.3»_«3o.357m question, -%2 2.ot49.61## -37.7»_«39.2s * - p<0.05 vs baseline; ** - p<0.01 vs vaseiine; *** - p<0.001 vs baseline## - p<0,01 vs ULD anti-PSA2 - shows decrease compared to the baseline in %, average group value The given data confirm that both ULD anti-PSA, and ULD anti-PSA + ULD anti-eNOS were used to effectively treat symptoms of lower urinary tract, increase averageand maximum urinary flow speed, improve quality of life of patients (Table 22). Thecourse of the was not long (12 weeks), therefore, a decrease in prostate volume was notobserved in any study group. ULD anti-PSA did not effect the volume of urination, whichincreased only in 52.6% patients, on average the group showed some statisticallyinsignificant decrease of urination volume by 11.8 ml (5.4%) compared to the baselinevalues. At the same time, patients, treated with ULD anti-PSA + ULD anti-eNOS,showed an increase in urination volume in 71.4%, and on average, an increase in volume was 48.3 ml (23.7%) compared to the baseline.

An analysis of dynamics of obstructive and irritative symptoms according to IPSSsubscales as well as nucturia evidence (question 7 of IPSS) showed that bothpharmaceutical compositions contributed to a decrease of obstruction and irritative symptoms, and also a decrease of nucturia symptoms. At the same time, a ULD anti-86 PSA +anti-eNOS was more effective compared to a ULD anti-PSA in decreasingirritative symptoms of lower urinary tract (28.2% vs. 40.3%, p<0.05) and nighttimeurination urges (2.0% vs. 37.7%,). lt should be noted, that ULD anti-PSA + ULD anti-eNOS is also more effectivecompared to ULD anti-PSA in improving erectile function in patients. ln ULD anti-PSA +ULD anti-eNOS group, the total ||EF (International Index of Erectile Dysfunction) scoreincreased by 19% in patients (in ULD anti-PSA group by 10.5%), an average increase of||EF score in ULD anti-PSA + ULD anti-eNOS group was 8% vs 4.5% in a ULD anti-PSA group.The pharmaceuticai compositions showed excellent safety profile, no adverse effects related to the administered medications were observed in the course of study.

Therefore, ULD anti-PSA + ULD anti-eNOS showed better efficacy compared tothat of ULD anti-PSA in treating urination problems caused by benign prostatichyperplasia. ln addition, a greater positive effect of ULD anti-PSA + ULD anti-eNOS onerectile function of patients compared to ULD anti-PSA was revealed. 87

Claims

1. OF PCTlIB2011/00g2_350 ïfïa-QMWA pharmaceutical composition comprising a) an activated-potentiated form of an antibody to an endogenous biological molecule, and b) an activated-potentiatedform of an antibody to NO synthase. šêlajåwThe pharmaceutical composition of claim 11, further comprisingpharmaceutically acceptable solid carrier. WThe pharmaceutical composition of claim 82, wherein said activated-potentiated form of an antibody to endothelial NO-synthase is in the form of a mixture ofof C12, C30, and C200 homeopathic dilutions impregnated onto said solid carrier. -*2--i¿š«,»§¿_,m,The pharmaceutical composition of claim ll, wherein said activated-potentiated form of an antibody to an endogenous biological molecule is a monoclonal, golyclonal, or natural antibody. ífê--t-uíšwwThe pharmaceutical composition of claim 495, wherein said antibody to an endogenous biological molecule is a polyclonal antibody.ss šwå--t-šywwThe pharmaceutical composition of claim 11, wherein said activated-potentiated form of an antibody to an endogenous biological molecule is prepared bysuccessive centesimal dilutions coupled with shaking of every dilution. éfâfzywWThe pharmaceutical composition of claims l_1_, wherein said antibody toendothelial NO-synthase is monoclonal, polyclonal or natural antibody. -få-rtßtåyuﬂmThe pharmaceutical composition of claim 82, wherein said antibody toendothelial NO-synthase is a polyclonal antibody. vš--ä-ïíuwThe pharmaceutical composition of claim Qå, the activated-potentiatedform of an antibody to endothelial NO-synthase is prepared by successive centesimaldilutions coupled with shaking of every dilution. 10.in increasinq the effect of the activated potentiated form of the antibodv to theendoqenous bioloqical molecule. 11. The pharmaceutical composition for use accordinq to claim 10 in a patientin need of treatment with said activated potentiated forrn of the antibodv. 12.activated potentiated form of the antibodv to the endoqenous bioloqical molecule is theantibodv to S-100 protein. 13.activated potentiated form of the antibodv to the endoqenous bioloqical molecule is theantibodv to prostate specific antiqen. 14. activated potentiated form of the antibodv to the endoqenous bioloßal molecule is the The pharmaceutical composition accordinq to one of claims 1 to 9 for use The pharmaceutical composition accordinq to claim 10 or 11, wherein said The pharmaceutical composition accordinq to claim 10 or 11 wherein said The pharmaceutical composition accordinq to claim 10 or 11 wherein said antibodv to insulin receptor.15. activated potentiated form of the antibodv to the endoqenous bioloqical molecule is the The pharmaceutical composition accordinq to claim 10 or 11 wherein said antibodv to anqiotensin receotor ll. 89