• 专利附录
  • 专利说明
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    • 专利摘要:
    The drug for the prophylaxis of HIV infection and for the prophylaxis and treatment of diseases caused by or associated with HIV, including AIDS, comprises an activated, potentiated form of antibodies to a protein or peptide of the immune system which interacts with the HIV or has a content and/or functional activity which changes in connection with an HIV infection. Furthermore, in the method for the prophylaxis of HIV infection and for the prophylaxis and treatment of diseases caused by or associated with HIV, including AIDS, use is made of an activated, potentiated form of antibodies to an antigen, namely a protein or peptide of the immune system, which interacts with the HIV or has a content and/or functional activity which changes in connection with an HIV infection.
    公开号:ES2555557A2
    申请号:ES201390021
    申请日:2011-07-15
    公开日:2016-01-04
    发明作者:Oleg Iliich Epshtein；Sergei Alexandrovich Tarasov
    申请人:Oleg Iliich Epshtein；
    IPC主号:
    专利说明:

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    DESCRIPTION
    Pharmacy and use to prepare a drug for the prevention of HIV infection and the treatment of HIV-caused diseases, including AIDS
    Technical field
    The invention belongs to the medical field and can be used to prepare a medicament intended for the effective prevention of HIV infection, the prevention and treatment of diseases caused by HIV, including AIDS.
    Prior art
    The prior art includes a drug for the treatment of infectious diseases, including viral diseases, based on the activated form of very small doses of anti-interferon antibodies (RU 2192888 C1, A61K39 / 395, 20.11.2002). However, this drug may not be effective in preventing HIV infection, nor in preventing and treating a broad spectrum of diseases caused by HIV or associated with HIV, including AIDS.
    Disclosure of the invention
    The invention is directed to the discovery of a complex drug without marked adverse effects, which allows the effective prevention of HIV infection as well! such as the prevention and effective treatment of diseases caused by HIV or associated with HIV, including infections and infestations, malignant neoplasms and AIDS in HIV-positive subjects.
    The solution to the problem posed is that the drug for the prevention of HIV infection, and the prevention and treatment of diseases caused by HIV or associated with HIV, including AIDS, according to the invention, contains a form activated - enhanced antibodies against an antigen - a protein or peptide of the immune system or, mainly, produced by the immune system, which interacts with HIV, or whose content and / or functional activity changes in the presence of HIV.
    It is also possible to use the activated-potentiated form of antibodies predominantly directed against the dissolved antigen (or against the soluble antigen, that is, the antigen, not bound to the outer membrane of the cells of the immune system).
    Cytokines (with the exception of interferon gamma) can be used as soluble antigens.
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    In addition, it is also possible to use the activated-enhanced form of antibodies directed predominantly against the antigen attached to the outer membrane of the cells of the immune system.
    For this, the immunocompetent cell receptors are used as antigen bound to the outer membrane of the cells of the immune system.
    In addition, as an antigen undio to the outer membrane of the cells of the immune system it is possible to use the differentiation clusters, (except for the CD4 molecules of the T lymphocytes).
    The activated-enhanced form of antibodies against dissolved antigens or the activated-enhanced form of antibodies against antigens bound to the outer membrane of the immune system cells is used in the form of an aqueous or aqueous-activated activated-potentiated solution whose activity is based in the multiple serial dilution process of the stock (initial) solution of the antibodies in an aqueous or aqueous-alcoholic solvent in combination with the external mechanical impact of vertical agitation after each dilution.
    For this, the claimed drug can be designed in solid dosage form of pharmaceutical composition, which contains the technologically necessary (effective) amount of the saturated neutral vehicle with the mixture of the aqueous or acylo-alcoholic solutions of the activated-potentiated forms of antibodies against dissolved antigens or the activated-enhanced form of antibodies against antigens bound to the outer membrane of the immune system cells, and pharmaceutically acceptable excipients, within which are, for example, lactose, microcrystalline cellulose and magnesium stearate.
    Aqueous or aqueous-alcoholic solutions of the activated-enhanced forms of antibodies against dissolved antigens or against antigens bound to the outer membrane of the immune system cells can be obtained by consecutive multiple dilution of the stock (initial) antibody solution. , in combination with the external mechanical impact of vertical agitation after each dilution. The concentration of the stock solution is 0.5 - 5.0 mg / ml.
    The activated-enhanced form of antibodies can be used in the form of a mixture of different dilutions, mainly centesimals, by homeopathic technology.
    The solution of the problem posed also ensures that in the mode of prevention of HIV infection and the effective treatment of diseases caused
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    by HIV or associated with HIV, including AIDS in accordance with the invention, the activated-potentiated form of antibodies against an antigen (protein or peptide of the immune system, or predominantly produced by the immune system) that reacts with HIV is used or whose content and / or functional activity is altered by HIV infection.
    The activated-enhanced form of antibodies against dissolved antigens or the activated-enhanced form of antibodies to antigens bound to the outer membrane of immune system cells are used in the form of activated aqueous or aqueous-alcoholic solution-enhanced of each component. , whose activity is conditioned by the multiple dilution process of the stock (initial) solution of antibodies in an aqueous or alcohol-alcoholic solvent in combination with the external mechanical impact of vertical agitation after each dilution.
    Solutions of the activated aqueous or aqueous-alcoholic forms - boosted of antibodies against dissolved antigens or against antigens bound to the outer membrane of the cells of the immune system are preferably obtained by means of serial serial dilution of the stock solution (initial ) of the antibodies, in combination with the external mechanical impact of vertical agitation after each dilution; The concentration of the stock solution is 0.5 - 5.0 mg / ml.
    According to the invention, the activated-potentiated form is an antibody form prepared according to the homeopathic potentiation technology by means of the serial multiple dilution of the stock (initial) antibody solution in combination of the external vertical agitation action for each dilution. , which has activity in the pharmacological models and / or in the climatic methods of prevention of HIV infection, prevention and treatment of diseases caused by HIV or diseases associated with HIV, including AIDS.
    The proposed use of the activated-potentiated form of antibodies against dissolved antigens (for example, against tumor necrosis factor alpha or human interferon alpha) or against antigens bound to the outer membrane of immune system cells (for example, against the CD8 receptor) leads to the unexpected therapeutic effect that consists in a greater efficiency of the drug, both in the prevention of HIV infection, and in the prevention and treatment of diseases caused by HIV or associated with HIV, including AIDS .
    It has been experimentally confirmed that the claimed drug is characterized by a high preventive efficacy with respect to HIV, preventing the penetration into human immunodeficiency virus cells and their intracellular replication and, thus, can be used
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    both for the effective treatment, as well as for the prevention of viral diseases with a tendency to chronic evolution, even for the secondary prevention of HIV infection.
    It is possible to use the claimed drug in combination with antiretroviral agents, including complex agents such as reverse transcriptase inhibitors (eg, zidovudine derivatives), which allows reducing the dose of antiretroviral agents while maintaining the high efficacy of therapy and reduce the rate of unwanted effects.
    Embodiments of the invention
    The drug is prepared, mainly, as follows.
    For the preparation of the activated-potentiated form of the active components, monoclonal or, mainly, polyclonal antibodies are used, which can be obtained by known technologies, in particular, the techniques described, for example, in the book: Immunological methods, in the edition G. Frimel, M., "Meditsyna", 1987, p. 9-33 [Russian]; or in the article by Laffly E., Sodoyer R. “Hum. Antibodies Monoclonal and recombinant antibodies, 30 years after ’- 2005 - Vol. 14. - N 1-2. P.33-55.
    Monoclonal antibodies are obtained, for example, by hybridoma technology. In it, the initial phase of the process includes immunization, based on principles already developed for the preparation of polyclonal antisera. The later stages of the process involve obtaining hybridoma cells, producing clones of antibodies of the same specificity. Their separation is done with the same methods that are used to obtain polyclonal antisera.
    Polyclonal antibodies can be obtained by active immunization of animals. For this, animals are administered, following a specially elaborated scheme, a series of injections of the required substance according to the invention - the antigen or the conjugated antigen (the protection or peptide of the immune system or, mainly produced by the immune system , which reacts with HIV or whose content and / or functional activity is altered by HIV infection). As a result of performing such a procedure, a monospedic antiserum with a high antibody content is obtained, which is used to obtain the activated-potentiated form. If necessary, purification of existing antibodies in the antiserum is performed, for example, by affinity chromatography, salt fractionation or ion exchange chromatography methods.
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    For example, polyclonal antibodies against tumor necrosis factor alpha can be used for the preparation of the claimed drug, which are used as a primary solution or stock (initial) (concentration of 0.5 - 5.0 mg / ml), for the subsequent preparation of the activated-enhanced form.
    For the preparation of the claimed drug, polyclonal antibodies are preferably used, which can be obtained by immunization of rabbits, as follows.
    For example, polyclonal antibodies against tumor necrosis factor alpha (TNF-a) can be obtained with the use of whole molecules of the TNF-a factor with the following sequence:
    1 MSTESMIRDV ELAEEALPKK TGGPQGSRRC LFLSLFSFLI VAGATTLFCL LHFGVIGPQR
    61 EEFPRDLSLI SPLAQAVRSS SRTPSDKPVA HVVANPQAEG QLQWLNRRAN ALLANGVELR
    121 DNQLVVPSEG LYLIYSQVLF KGQGCPSTHV LLTHTISRIA VSYQTKVNLL
    SAIKSPCQRE
    181 TPEGAEAKPW YEPIYLGGVF QLEKGDRLSA EINRPDYLDF AESGQVYFGIIAL
    To obtain polyclonal antibodies against tumor necrosis factor-alpha (TNF-a) it is possible to use a polypeptide fragment of tumor necrosis factor alpha selected, for example, from the following sequences:
    PSDKP
    93-97:
    VANPQ
    65-199:
    RDLSLI SPLAQAVRSS SRTPSDKPVA HVVANPQAEG QLQWLNRRAN ALLANGVELR DNQLVVPSEG LYLIYSQVLL KGQGCPSTHV LLTHTISRIA VSYQTKVNLL SAIKSPCQRE TPEGAEAKPW YEPIYLGG
    77-93:
    RSS SRTPSDKPVA HVV
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    32-54:
    GGPQGSRRC LFLSLFSFLI VAGA 56-73:
    IGPQR EEFPRDLSLI SPL 123-160:
    QLVVPSEG LYLIYSQVLF KGQGCPSTHV LLTHTISRIA 176-190:
    PCQRE TPEGAEAKPW 5- 45:
    SMIRDV ELAEEALPKK TGGPQGSRRC LFLSL 150-184:
    V LLTHTISRIA VSYQTKVNLL SAIKSPCQRE TPEG 77-233:
    VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGV
    ELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQ
    TKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSA
    EINRPDYLDFAESGQVYFGIIAL.
    Before blood collection, 1-3 intravenous injections are performed for 7-9 days to increase the level of antibodies. During the immunization process, rabbits are taken small blood samples to assess the amount of antibodies. The maximum level of immune response to the administration of most antigens is reached in 40-60 days after the first injection. After the end of the first cycle of immunization of rabbits, their health is allowed to be restored for 30 days and the immunization is performed, which includes 1-3 intravenous injections. To obtain the antiserum from the immunized rabbits, blood is collected in a 50 ml volume centrifuge tube. With the help of a wooden spatula, the clots that have formed are removed from the tube walls and the spatula is placed in the clot formed in the center of the tube. Blood is placed in the refrigerator (temperature 4 ° C) overnight. The next day the clot that has stuck to the spatula is removed, and the remaining liquid is centrifuged at 13000g for 10
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    minutes The supernatant (the Kid over the precipitate) is the antiserum. The antiserum obtained must be yellow. 20% NaN3 can be added to the antiserum until the final concentration of 0.02% and it should be kept frozen until its use at - 20 ° C temperature (or without adding NaN3 at -70 ° C temperature). The separation of antibodies from the antiserum against tumor necrosis factor - alpha can be done as follows:
    1. Dissolve 10 ml of the rabbit antiserum in 2 times 0.15 M NaCl, add 6.26 g of Na2SO4, mix and incubate for 12-16 h at 4 ° C;
    2. The precipitate formed is removed by centrifugation, dissolved in 10 ml of phosphate buffer and then dialyzed against the same buffer overnight at room temperature;
    3. After removing the sediment by centrifugation, the solution is taken to the column with DEAE-cellulose, equilibrated with the phosphate buffer;
    4. The fraction of antibodies is determined by measuring the optical density of the eluate at 280 nm.
    The purification of the antibodies is carried out by the column affinity chromatography method with the antigen, by means of the union of the antibodies against the tumor necrosis factor alpha with the antigen (tumor necrosis factor alpha), connected to the insoluble matrix of the column, with its subsequent elution of the antibodies by concentrated salt solutions.
    The buffer solution obtained in this way, from polyclonal antibodies against tumor necrosis factor alpha with a concentration of 0.5 - 5.0 mg / ml, preferably 2.0 + 3.0 mg / ml, is used as the matrix solution (initial ) for the subsequent preparation of the activated-enhanced form of the antibodies.
    Polyclonal antibodies against human alpha interferon are obtained by the method indicated above, using as adjuvant immunogen (antigen) for the immunization of rabbits and the entire molecule of human interferon alpha from one of the following sequences:
    Interferon alfa human (subtype 2):
    MALTFALLVA LLVLSCKSSC SVGCDLPQTH SLGSRRTLML LAQMRKISLF SCLKDRHDFG
    FPQEEFGNQF QKAETIPVLH EMIQQIFNLF STKDSSAAWD ETLLDKFYTE LYQQLNDLEA
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    CVIQGVGVTE TPLMKEDSIL AVRKYFQRIT LYLKEKKYSP
    CAWEVVRAEIMRSFSLSTNL
    QESLRSKE
    Interferon alfa human (subtype 1/13): MASPFALLMV LVVLSCKSSC SLGCDLPETH SLDNRRTLML L AQMSRISPS SCLMDRHDFG
    FPQEEFDGNQ FQKAPAISVL HELIQQIFNL FTTKDSSAAW DEDLLDKFCT ELYQQLNDLE
    ACVMQEERVG ETPLMNADSILAVKKYFRRI TLYLTEKKYS
    PCAWEVVRAE IMRSLSLSTN
    LQERLRRKE
    Human alpha interferon (subtype 17):
    MALSFSLLMA VLVLSYKSIC SLGCDLPQTH SLGNRRALIL LAQMGRISPF SCLKDRHDFG
    LPQEEFDGNQ FQKTQAISVL HEMIQQTFNL FSTEDSSAAW EQSLLEKFST ELYQQLNNLE
    ACVIQEVGME ETPLMNEDSI LAVRKYFQRI TLYLTEKKYS
    PCAWEVVRAEIMRSLSFSTN
    LQKILRRKD
    Human alpha interferon (subtype 4):
    MALSFSLLMA VLVLSYKSIC SLGCDLPQTH SLGNRRALIL LAQMGRISHF SCLKDRHDFG
    FPEEEFDGHQ FQKAQAISVL HEMIQQTFNL FSTEDSSAAW EQSLLEKFST ELYQQLNDLE
    ACVIQEVGVE ETPLMNEDSI LAVRKYFQRI TLYLTEKKYS
    PCAWEVVRAE IMRSLSFSTN
    LQKRLRRKD
    Human alpha interferon (subtype 8):
    MALTFYLLVA LVVLSYKSFS SLGCDLPQTH SLGNRRALIL LAQMRRISPF SCLKDRHDFE
    FPQEEFDDKQ FQKAQAISVL HEMIQQTFNL FSTKDSSAAL DETLLDEFYIELDQQLNDLE
    SCVMQEVGVI ESPLMYEDSI LAVRKYFQRI TLYLTEKKYS
    SCAWEVVRAE IMRSFSLSIN
    LQKRLKSKE
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    Interferon alfa human (subtype 7):
    MARSFSLLMV VLVLSYKSIC SLGCDLPQTH SLRNRRALIL LAQMGRISPF SCLKDRHEFR
    FPEEEFDGHQ FQKTQAISVL HEMIQQTFNL FSTEDSSAAW EQSLLEKFST ELYQQLNDLE
    ACVIQEVGVE ETPLMNEDFI LAVRKYFQRI TLYLMEKKYS
    PCAWEVVRAE IMRSFSFSTN
    LKKGLRRKD
    Human alpha interferon (subtype 21):
    MALSFSLLMA VLVLSYKSIC SLGCDLPQTH SLGNRRALIL LAQMGRISPF SCLKDRHDFG
    FPQEEFDGNQ FQKAQAISVL HEMIQQTFNL FSTKDSSATW EQSLLEKFST ELNQQLNDLE
    ACVIQEVGVE ETPLMNVDSI LAVKKYFQRI TLYLTEKKYS
    PCAWEVVRAE IMRSFSLSKI
    FQERLRRKE
    Human alpha interferon (subtype 10):
    MALSFSLLMA VLVLSYKSIC SLGCDLPQTH SLGNRRALIL LGQMGRISPF SCLKDRHDFR
    IPQEEFDGNQ FQKAQAISVL HEMIQQTFNL FSTEDSSAAW EQSLLEKFST ELYQQLNDLE
    ACVIQEVGVE ETPLMNEDSI LAVRKYFQRI TLYLIERKYS
    PCAWEVVRAE IMRSLSFSTN
    LQKRLRRKD
    Human alpha interferon (subtype 14):
    MALPFALMMA LVVLSCKSSC SLGCNLSQTH SLNNRRTLML MAQMRRISPF SCLKDRHDFE
    FPQEEFDGNQ FQKAQAISVL HEMMQQTFNL FSTKNSSAAW DETLLEKFYIELFQQMNDLE
    ACVIQEVGVE ETPLMNEDSI LAVKKYFQRI TLYLMEKKYS
    PCAWEVVRAE IMRSLSFSTN
    LQKRLRRKD
    Interferon alfa human (subtype 5):
    MALPFVLLMA LVVLNCKSIC SLGCDLPQTH SLSNRRTLMI MAQMGRISPF SCLKDRHDFG
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    FPQEEFDGNQ FQKAQAISVL HEMIQQTFNL FSTKDSSATW DETLLDKFYT ELYQQLNDLE
    ACMMQEVGVE DTPLMNVDSILTVRKYFQRI TLYLTEKKYS
    PCAWEVVRAE IMRSFSLSAN
    LQERLRRKE
    The use of adjuvant and, for example, a polypeptide fragment of human interferon alpha, such as the immunogen (antigen) for the immunization of rabbits, is possible for obtaining polyclonal antibodies against human interferon alfa.
    Polyclonal antibodies against the CD8 receptor are obtained by the method indicated above, using an adjuvant and the entire molecule of the CD8 receptor with the following amino acid sequence as immunogen (antigen) for rabbit immunization:
    1 MALPVTALLL PLALLLHAAR PSQFRVSPLD RTWNLGETVE LKCQVLLSNP TSGCSWLFQP
    61 RGAAASPTFL LYLSQNKPKA AEGLDTQRFS GKRLGDTFVL TLSDFRRENE GYYFCSALSN
    121 SIMYFSHFVP VFLPAKPTTT PAPRPPTPAP TIASQPLSLR PEACRPAAGG AVHTRGLDFA
    181 CDIYIWAPLA GTCGVLLLSL VITLYCNHRN RRRVCKCPRP VVKSGDKPSL SARYV.
    To obtain polyclonal antibodies against the CD8 receptor, it is possible to use, as immunogen (antigen) for the immunization of rabbits, an adjuvant and, for example, a polypeptide fragment of the CD8 receptor chosen from the following sequences:
    11-30:
    PLALLLHAAR PSQFRVSPLD;
    81-100:
    AEGLDTQRFS GKRLGDTFVL;
    121-140:
    SIMYFSHFVP VFLPAKPTTT;
    201-210:
    VITLYCNHRN;
    221-235:
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    VVKSGDKPSL SARYV
    For the preparation of the drug, preferably the mixture of three aqueous-alcoholic dilutions of the primary matrix solution of the antibodies diluted 10012, 10030 and 100200 times, respectively, is used, which corresponds to the centesimal dilutions C12, C30 and C200, prepared by homeopathic technology. During the realization of the claimed drug in its medicinal solid form, the mixture of indicated components is taken to a neutral vehicle.
    The activated-enhanced form of each component is prepared by uniform reduction of the concentration as a result of the consecutive dilution of 1 part of each solution under dilution, starting with the mentioned matrix solution, in 9 parts (for the decimal dilution D) or in 99 parts (for the centesimal dilution C) or in 999 parts (for the milimal dilution M) of the neutral solvent in combination of the repeated vertical agitation (potentiation, or "dynamization") of each dilution obtained and the use of independent containers for each consecutive dilution until the necessary potency is obtained - multiplicity of the dilution by the homeopathic method (see, for example, W. Schwabe "Homeopathic medicinal preparations", M, 1967, p. 14-29).
    External treatment during the concentration reduction process can also be performed by ultrasound, electromagnetism or any other physical action.
    For example, for the preparation of the 12th C12 centesimal dilution, a part of the matrix (primary) solution of the antibodies is diluted, for example against the tumor necrosis factor alpha with the concentration of 2.5 mg / ml in 99 parts of neutral water or of an aqueous-alcoholic solvent and is stirred vertically repeatedly (10 and more times) - the potentiation occurs, obtaining the first centesimal dilution C1. From this 1st centesimal dilution C1 the 2nd centesimal dilution C2 is prepared. This operation is repeated 11 times, obtaining the 12th C12 centesimal dilution. Thus, the 12th C12 centesimal dilution is the solution obtained by consecutive dilution by diluting one part of the primary matrix solution, which is the solution of antibodies against human gamma interferon with the concentration of 2.5 mg / ml, in 99 parts of neutral solvent 12 consecutive times in different containers, ie the solution obtained by diluting the matrix solution 10012 times. Similar operations are carried out with the multiplicity of corresponding dilutions to obtain dilutions C30 and C 50.
    In the use, for example, of the activated-enhanced form of antibodies against tumor necrosis factor-alpha in the form of a mixture of different dilutions, mainly
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    centesimals, each dilution (for example, C 12, C30, C50) is prepared individually by the technology described above until the dilution of 3 dilutions less than the final one (for obtaining C9, C27, C47, respectively), and then Take a part of each component, according to the composition of the mixture, to a container and mix with the necessary amount of solvent (with 97 parts for centesimal dilution, respectively). Then the mixture obtained is diluted twice consecutively in the proportion of 1 to 100, enhancing the solution obtained after each dilution. This results in the activated-enhanced form of the antibodies, for example, against human gamma interferon in ultra-low doses, obtained by diluting the matrix solution 10012, 10030, 10050 times, which is equivalent to the dilution mixture. centesimals C12, C30, C50.
    It is possible to use each component separately in the form of a mixture of other different dilutions, for example, decimals and / or centesimals, (D 20, C 30, C 100 or C12, C30, C200, etc.), prepared by technology homeopathic, whose effectiveness is determined experimentally.
    In order to obtain the solid form of oral administration of the claimed drug, the irrigation process is carried out in the fluidized bed installation (for example, of the type of "Huttlin Pilotlab" produced by the company Huttlin GmbH) until saturation of the granules of the substance neutral that are injected into the boiling pseudo-fluidized bed; the substance is lactose (milk sugar) with 50 * 500 ^ m of particle size, previously obtained by means of the aqueous or aqueous-alcoholic solution of the activated-enhanced form of the antibodies against the CD4 receptor, mainly in proportion of 1 kg of antibody solution in 5 or 10 kg of lactose (1: 5 - 1:10) with simultaneous drying in the flow of hot air supplied under the rack at a temperature not exceeding 40 ° C. The calculated amount of lactose (10 * 91% of the tablet mass), saturated with the activated form - boosted of the antibodies by the method indicated above, is loaded into the mixer and mixed with the lactose, moistened with the activated form. - enhanced of the antibodies, in the amount of 3 - 10% of the mass of the tablet and with the pure lactose in an amount not exceeding 84% of the mass of the tablet (to reduce costs and simplify and accelerate the technological process without reducing medicinal efficacy). Then, the microcrystalline cellulose in the amount of 5 + 10% of the mass of the tablet and magnesium stearate in the amount of 1% of the mass of the tablet are added to the mixture. The mass of tablet obtained is uniformly mixed and compressed by direct dry compression (for example, in the Korsch-XL 400 compressor). After tablet formation
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    300 mg tablets of saturated mass of the aqueous-alcoholic solution are obtained in the activated-enhanced form of the antibodies against the tumor necrosis factor alpha in ultra-low doses of each component prepared from the matrix solution, diluted 10012, 10030, 10050 times, which is equivalent to the mixture of C12, C30 and C50 centesimal dilutions, prepared according to homeopathic technology.
    It is recommended to preferably take 1-2 tablets of the claimed drug 2-4 times a day.
    Example 1.
    The antiretroviral action of the claimed drug was studied, inhibiting HIV replication in the culture of human peripheral blood mononuclear cells, infected in vitro with the strain HIV-1-LAI. The efficacy of HIV replication inhibition was assessed by the content of the basic HIV nucleocapside protein p24 in supernatants.
    For the realization of the experimental studies, affinity purified anti-tumor necrosis factor anti-tumor necrosis factor rabbit polyclonal antibodies were used, prepared on request by a specialized biotechnology company, which were the basis for preparing the activated-enhanced form of aqueous dilutions of antibodies against the tumor necrosis factor alpha in the ultra-low dose, obtained using homeopathic technology by superdilution of the primary matrix solution (concentration of 2.5 mg / ml) 10012, 10030, 10050 times, which is equivalent to the mixture of centesimal homeopathic dilutions C12, C30, C50 (hereinafter: DUB AC anti-TNF alpha). The evaluation of the antivrnca activity of the prepared complex was carried out with the use of human peripheral blood mononuclear cells that were infected in vitro with the strain HIV-1-LAI.
    The human peripheral blood mononuclear cells were isolated from the blood of a healthy seronegative donor by centrifugation in a Ficoll-Hypaque density gradient. The cells were stimulated for 3 days with the use of 1 ^ g / ml of phytohemagglutinin P and 5 Ul / ml of recombinant human interleukin.
    In order to evaluate the antiretroviral activity, the preparations were placed in a well containing 100 µl of activated human peripheral blood mononuclear cells, either 24 hours before or 15 minutes after infection of the cells with the HIV-1 strain -LAI at a dose of 100 TCID50 (50 ^ l inoculum of the strain HIV-1-LAI). Before placing it in the well, DUB AC anti-TNF-alpha (12.5 ^ l) or azidothymidine (active substance
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    - zidovudine) at a concentration of 1000 nM (the preparation control) was mixed with RPMI1640 medium (DIFCO) until reaching the final volume of 50 ^ l.
    Supernatants from cell cultures were collected on day 7 after infection of the cells. The efficacy of the products was defined from the inhibition of HIV replication, which was evaluated by the content of the basic p24 HIV nucleocapside protein in cell supernatants using the ELISA method (Retrotek ELISA kit ).
    It has been found that DUB AC anti-TNF-alpha inhibits HIV replication in 92 ± 3% when added to the container 24 hours before, and in 13 ± 13% when added to the container 15 minutes after infection of the cells with the strain HIV-1-LAI, respectively. Azidothymidine in the 1000 nM dose inhibited HIV replication in 99 ± 0 and 99 ± 1%, respectively, when it was added 24 hours before and 15 minutes after infection of the cells with the HIV-1-LAI strain.
    Thus, the in vitro study has shown the high antiretroviral activity of the claimed preparation based on the activated-enhanced form of rabbit polyclonal antibodies against TNF alpha.
    Note: TCID50 is the dose that infects 50% of tissue culture cells.
    Example 2
    The antiretroviral action of the claimed drug was studied, inhibiting HIV replication in the culture of human peripheral blood mononuclear cells, infected in vitro with the strain HIV-1-LAI. The efficacy of HIV replication inhibition was assessed by the basic protein content of HIV p24 nucleocapside in supernatants.
    For the realization of the experimental studies, affinity purified rabbit polyclonal antibodies against CD8 were used, commissioned by a specialized biotechnology company, which were the basis for preparing the activated-enhanced form of aqueous dilutions of antibodies against CD8 in ultra-low doses, obtained using homeopathic technology by super-dilution of the primary matrix solution (concentration of 2.5 mg / ml) 10012, 10030, 10050 times, which is equivalent to the mixture of the homeopathic dilutions centesimals C12, C30, C50 (hereinafter: DUB AC CD8).
    The evaluation of the antivmca activity of the prepared complex was carried out with the use of
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    human peripheral blood mononuclear cells that were infected in vitro with the strain HIV-1-LAI. Human peripheral blood mononuclear cells were isolated from the blood of a healthy seronegative donor by centrifugation in a Ficoll-Hypaque density gradient. The cells were stimulated for 3 days with the use of 1 ^ g / ml of phytohemagglutinin P and 5 Ul / ml of recombinant human interleukin-2.
    In order to evaluate the antiretroviral activity, the preparations were placed in a well containing 100 ^ l of activated human peripheral blood mononuclear cells, either 24 hours before or 15 minutes after infection of the cells with the HIV-1 strain -LAI at a dose of 100 TCID50 (50 ^ l inoculum of the strain HIV-1-LAI). Before placing it in the well, DUB AC CD8 (12.5 ^ l) or azidothymidine (active substance - zidovudine) at a concentration of 1000 nM (the preparation control) was mixed with RPMI1640 medium (DIFCO) until the final volume was reached of 50 ^ l.
    The cell culture supernatants were collected on day 7 after cell infection. The efficacy of the products was defined from the inhibition of HIV replication, which was evaluated by the content of the basic p24 HIV nucleocapside protein in cell supernatants using the ELISA method (Retrotek ELISA kit ).
    It has been shown that DUB AC CD8 inhibits HIV replication in 87 ± 11% when added to the container 24 hours before, and in 40 ± 4% when added to the container 15 minutes after infection of the cells with the strain HIV-1-LAI, respectively. Azidothymidine in the 1000 nM dose inhibited HIV replication in 99 ± 0 and 99 ± 1%, respectively, when it was added 24 hours before and 15 minutes after infection of the cells with the HIV-1-LAI strain.
    Thus, the in vitro study has shown a high antiretroviral activity of ultra low doses of rabbit polyclonal antibodies against CD8.
    Note: TCID50 - dose that infects 50% of tissue culture cells.
    Example 3
    The antiretroviral activity of the activated-potency forms of aqueous dilutions of affinity purified rabbit polyclonal antibodies directed against ultra-low dose interferon alfa (DUB), obtained using homeopathic technology by superdilution of the primary matrix solution (concentration of 2 , 5 mg / ml) 10012, 10030, 10050 times, which is equivalent to the mixture of homeopathic dilutions
    5
    10
    fifteen
    twenty
    25
    30
    Centesimals C12, C30, C50 (hereinafter: DUB AC anti-IFN-a) was evaluated with the use of human peripheral blood mononuclear cells that were infected in vitro with the strain HIV-1-LAI. As preparation control azidothymidine was used.
    The human peripheral blood mononuclear cells were isolated from the blood of a healthy seronegative donor by centrifugation in a Ficoll-Hypaque density gradient. The cells were stimulated for 3 days with the use of 1 ^ g / ml of phytohemagglutinin P and 5 Ul / ml of recombinant human interleukin-2.
    The cells were infected with the HIV-1-LAI strain, placing 50 ^ l inoculum of the HIV-1-LAI strain corresponding to a dose of 100 TCID50 (dose that infects 50% of the tissue culture cells ).
    In order to evaluate the antiretroviral activity, the DUB AC anti-IFN-a preparation and the azidothymidine control preparation were placed in a well containing 100 µl of activated human peripheral blood mononuclear cells, either 24 hours before or 15 minutes after infection of the cells with the strain HIV-1-LAI. Before placing it in the well, the preparation of DUB AC anti-IFN-a (12.5 ^ l) and the preparation of azidothymidine at a concentration 1000 nM were mixed with RPMI1640 medium (DIFCO) until reaching the final volume of 50 ^ l.
    Supernatants from cell cultures were collected on day 7 after infection. The efficacy of the preparations was defined from the inhibition of HIV replication, which was evaluated by the content of the basic nucleocapside HIV p24 protein in cell supernatants using the ELISA method (Retrotek ELISA kit ).
    It has been shown that DUB AC anti-IFN-a inhibits HIV replication in 95 ± 2% when added to the container 24 hours before, and in 59 ± 14% when added to the container 15 minutes after infection of the cells with the strain HIV-1-LAI, respectively. Azidothymidine in the 1000 nM dose inhibited HIV replication in 99 ± 0 and 99 ± 1%, respectively, when it was added 24 hours before and 15 minutes after infection of the cells with the HIV-1-LAI strain.
    Thus, the in vitro study has shown the high antiretroviral activity of the DUB AC anti-IFN-a preparation.
    权利要求:
    Claims (7)
    [1]
    5
    10
    fifteen
    twenty
    25
    30

    1. A drug characterized in that it contains the activated-potentiated form of antibodies against a protelna or peptide of the immune system that interacts with HIV or whose content and / or functional activity changes due to HIV infection, which is the necrosis factor alpha tumor
    [2]
    2. The drug according to claim 1, characterized in that it is in solid form of pharmaceutical composition, which contains a technologically necessary amount of saturated neutral vehicle with the mixture of aqueous or aqueous-alcoholic solutions of the activated-potentiated form of the antibodies. against tumor necrosis factor alpha, and acceptable pharmaceutical excipients.
    [3]
    3. The drug according to claim 2, characterized in that the acceptable pharmaceutical excipients include lactose, microcrystalline cellulose and magnesium stearate.
    [4]
    4. A method for the preparation of a drug according to claim 1, characterized in that the activated-enhanced form of the antibodies against the tumor necrosis factor alpha, is prepared in the form of an aqueous or aqueous-activated-enhanced alcoholic solution, whose activity is obtained by the process of multiple consecutive dilution of the initial matrix solution of the antibodies in an aqueous or aqueous-alcoholic solvent in combination with the external mechanical action of vertical agitation of each dilution.
    [5]
    5. The method according to claim 4, characterized in that the aqueous or aqueous-alcoholic solutions of the activated-potentiated forms of the antibodies against the tumor necrosis factor alpha are obtained by means of the consecutive multiple dilution of the initial matrix solution of antibodies, in combination with the external mechanical impact of vertical agitation after each dilution, where the concentration of the matrix solution is 0.5 - 5.0 mg / ml.
    [6]
    6. The method according to claim 5, characterized in that the aqueous or aqueous-alcoholic solutions of the activated-potentiated forms of the antibodies are prepared as a mixture of different dilutions, mainly centesimals, prepared by homeopathic technology.
    [7]
    7. Use of a drug of any one of claims 1 to 3, to prepare a medicament intended for the prevention of HIV infection and for the treatment of HIV diseases, including AIDS.
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    CA2807540A1|2012-02-09|
    RU2010133045A|2012-02-20|
    WO2012018284A1|2012-02-09|
    EP2601967A1|2013-06-12|
    AU2011286486B9|2017-02-23|
    DE112011102649T5|2013-06-20|
    GB2498276A|2013-07-10|
    AU2011286486A2|2013-05-02|
    GB201303983D0|2013-04-17|
    RU2535033C2|2014-12-10|
    UA112747C2|2016-10-25|
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    DE4236069A1|1992-10-26|1994-04-28|Werner Dr Bergmann|Homeopathic medicaments for treating HIV infections - prepd. by dilution of infected serum|
    US20030228310A1|1996-12-23|2003-12-11|Advanced Biotherapy, Inc.|Treatment of skin diseases|
    NZ284214A|1994-03-31|1997-07-27|Biomed Comm Inc|Extreme dilutions of growth factors, homeopathic compositions, radiotherapy.|
    US6090388A|1998-06-20|2000-07-18|United Biomedical Inc.|Peptide composition for prevention and treatment of HIV infection and immune disorders|
    JP2000143537A|1998-11-13|2000-05-23|Nippon Zoki Pharmaceut Co Ltd|Agent for suppressing expression of cell adhesion molecule|
    RU2181297C2|2000-06-20|2002-04-20|Эпштейн Олег Ильич|Method of treatment of pathological syndrome and medicinal agent|
    RU2192888C1|2001-02-15|2002-11-20|Эпштейн Олег Ильич|Medicinal agent and method of treatment of pathological syndrome|
    RU2195317C1|2001-04-18|2002-12-27|Эпштейн Олег Ильич|Method of correcting pathologic immune reactions and therapeutic agent|
    RU2201255C1|2001-12-26|2003-03-27|Эпштейн Олег Ильич|Medicinal agent and method of regulation of vascular tonus|
    RU2001134982A|2001-12-26|2004-02-20|Олег Ильич Эпштейн|METHOD OF IMMUNE RESPONSE CORRECTION AND MEDICINE|
    RU2205025C1|2001-12-26|2003-05-27|Гольдберг Евгений Данилович|Method of correction of immune response and medicinal agent|
    US20060153845A1|2002-08-02|2006-07-13|Epshtein Oleg I|Method for correcting immune respones and medical agent|
    JP2009504686A|2005-08-15|2009-02-05|アラーナ・テラピューティクス・リミテッド|Chimeric antibodies using the New World primate domain|
    US7622116B2|2006-02-10|2009-11-24|Zymogenetics, Inc.|Method of treating inflammation using soluble IL-17RCX4|
    RU2309732C1|2006-03-13|2007-11-10|Олег Ильич Эпштейн|Pressed solid oral formulation of medicinal preparation and method for preparing solid oral formulation of medicinal preparation|
    ES2687259T3|2008-06-25|2018-10-24|Esbatech, An Alcon Biomedical Research Unit Llc|Stable and soluble antibodies that inhibit TNF|
    CN103154030A|2010-08-06|2013-06-12|奥列格·伊里奇·爱泼斯坦|Combination pharmaceutical composition and methods of treating and preventing the infectious diseases|RU2181297C2|2000-06-20|2002-04-20|Эпштейн Олег Ильич|Method of treatment of pathological syndrome and medicinal agent|
    RU2309732C1|2006-03-13|2007-11-10|Олег Ильич Эпштейн|Pressed solid oral formulation of medicinal preparation and method for preparing solid oral formulation of medicinal preparation|
    DE112011102358T5|2010-07-15|2013-04-25|Oleg Iliich Epshtein|A method of increasing the effect of an activated-potentiated form of an antibody|
    US8637034B2|2010-07-15|2014-01-28|Oleg I. Epshtein|Pharmaceutical compositions comprising activated-potentiated antibodies to interferon-gamma and S100 protein|
    UA112755C2|2010-07-21|2016-10-25|Олєг Ільіч Епштейн|A method of treating attention deficit hyperactivity disorder|
    WO2014116789A1|2013-01-25|2014-07-31|Thymon, Llc|Immunogenic and prophylactic compositions, methods of making same, and method for treating and preventing tnf-mediated disease and hiv-1 infection|
    WO2014123696A1|2013-01-25|2014-08-14|Thymon, Llc|Compositions for selective reduction of circulating bioactive soluble tnf and methods for treating tnf-mediated disease|
    RU2013111962A|2013-03-18|2014-09-27|Олег Ильич Эпштейн|METHOD FOR DETERMINING THE EXPRESSION OF MODIFICATION ACTIVITY ASSOCIATED WITH A CARRIER|
    RU2013111961A|2013-03-18|2014-09-27|Олег Ильич Эпштейн|METHOD FOR DETERMINING THE EXPRESSION OF MODIFICATION ACTIVITY ASSOCIATED WITH A CARRIER|
    法律状态:
    2018-02-28| FC2A| Grant refused|Effective date: 20180222 |
    优先权:
    申请号 | 申请日 | 专利标题
    RU2010133045|2010-08-06|
    RU2010133045/15A|RU2535033C2|2010-08-06|2010-08-06|Therapeutic agent and method for prevention of hiv infection and treatment of hiv-caused or hiv-associated diseases, including aids|
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