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RU95117922A - COMPOSITION AND METHODS OF DELIVERY OF GENETIC MATERIAL - Google Patents

COMPOSITION AND METHODS OF DELIVERY OF GENETIC MATERIAL

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Publication number
RU95117922A
RU95117922A RU95117922/14A RU95117922A RU95117922A RU 95117922 A RU95117922 A RU 95117922A RU 95117922/14 A RU95117922/14 A RU 95117922/14A RU 95117922 A RU95117922 A RU 95117922A RU 95117922 A RU95117922 A RU 95117922A
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Russia
Prior art keywords
nucleic acid
hiv
acid molecule
sequence encoding
virus
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RU95117922/14A
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Russian (ru)
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RU2174845C2 (en
Inventor
Б.Вейнер Дэвид
В.Вилльямс Вилльям
ВАНГ Бин
Р.Кони Лесли
Дж.Мерва Майкл
Р.Зуравски Винсент (младший)
Original Assignee
Б.Вейнер Дэвид
В.Вилльямс Вилльям
ВАНГ Бин
Р.Кони Лесли
Дж.Мерва Майкл
Р.Зуравски Винсент (младший)
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Publication of RU95117922A publication Critical patent/RU95117922A/en
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Publication of RU2174845C2 publication Critical patent/RU2174845C2/en

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Claims (48)

1. Способ введения в клетки индивидуума генетического материала, отличающийся тем, что осуществляют: а) контактирование клеток индивидуума с усилителем функции полинуклеотида; b) введение в клетки индивидуума молекулы нуклеиновой кислоты, причем молекула нуклеиновой кислоты не содержит частиц ретровируса.1. The method of introducing into the cells of an individual genetic material, characterized in that they carry out: a) contacting the cells of the individual with an enhancer of polynucleotide function; b) introducing into the cells of an individual a nucleic acid molecule, wherein the nucleic acid molecule does not contain retrovirus particles. 2. Способ по п. 1, отличающийся тем, что усилитель функции полинуклеотида представлен бупивакаином. 2. The method according to p. 1, characterized in that the polynucleotide function enhancer is represented by bupivacaine. 3. Способ по п. 1, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок и операбельно связанную с регуляторными последовательностями. 3. The method according to p. 1, characterized in that the nucleic acid molecule comprises a nucleotide sequence encoding a protein and operably linked to regulatory sequences. 4. Способ по п. 1, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок, в состав которого входит по меньшей мере один эпитоп, идентичный или по существу аналогичный эпитопу антигена, против которого желателен иммунный ответ, причем нуклеотидная последовательность операбельно связана с регуляторными последовательностями. 4. The method according to p. 1, characterized in that the nucleic acid molecule comprises a nucleotide sequence encoding a protein, which includes at least one epitope that is identical or substantially similar to the epitope of the antigen against which an immune response is desired, the nucleotide sequence being operable linked to regulatory sequences. 5. Способ по п. 1, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок, в состав которого входит по крайней мере один эпитоп, идентичный или по существу подобный эпитопу антигена патогена. 5. The method of claim 1, wherein the nucleic acid molecule comprises a nucleotide sequence encoding a protein that comprises at least one epitope that is identical or substantially similar to an epitope of a pathogen antigen. 6. Способ по п. 5, отличающийся тем, что усилитель функции полинуклеотида представлен бупивакаином. 6. The method according to p. 5, characterized in that the polynucleotide function enhancer is represented by bupivacaine. 7. Способ по п. 5, отличающийся тем, что молекула нуклеиновой кислоты представлена молекулой ДНК. 7. The method according to p. 5, characterized in that the nucleic acid molecule is a DNA molecule. 8. Способ по п. 5, отличающийся тем, что белок представлен антигеном патогена или его фрагментом. 8. The method according to p. 5, characterized in that the protein is represented by a pathogen antigen or fragment thereof. 9. Способ по п. 5, отличающийся тем, что молекулу нуклеиновой кислоты вводят внутримышечно. 9. The method according to p. 5, characterized in that the nucleic acid molecule is administered intramuscularly. 10. Способ по п. 5, отличающийся тем, что патоген представлен вирусом, выбранным из группы, включающей вирус иммунодефицита человека (ВИЧ); вирус Т-клеточного лейкоза человека (TPLV), вирус гриппа, вирус гепатита A (HAV), вирус гепатита В (HBV), вирус гепатита С (HCV), вирус папилломы человека (HPV), вирус простого герпеса 1 (HSV1), вирус простого герпеса 2 (HSV2), цитомегаловирус (CMV), вирус Энштейна-Барра (TBV), риновирус и коронавирус. 10. The method according to p. 5, characterized in that the pathogen is a virus selected from the group comprising the human immunodeficiency virus (HIV); human T-cell leukemia virus (TPLV), influenza virus, hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), human papillomavirus (HPV), herpes simplex virus 1 (HSV1), virus herpes simplex 2 (HSV2), cytomegalovirus (CMV), Einstein-Barr virus (TBV), rhinovirus and coronavirus. 11. Способ по п. 5, отличающийся тем, что патоген представлен ВИЧ и молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок ВИЧ. 11. The method according to p. 5, characterized in that the pathogen is HIV and the nucleic acid molecule comprises a nucleotide sequence encoding an HIV protein. 12. Способ по п. 5, отличающийся тем, что патоген представлен ВИЧ и молекула нуклеиновой кислоты включает нуклеотидные последовательности, кодирующие более одного структурного белка ВИЧ. 12. The method according to p. 5, characterized in that the pathogen is represented by HIV and the nucleic acid molecule includes nucleotide sequences encoding more than one structural protein of HIV. 13. Способ по п. 5, отличающийся тем, что патоген представлен ВИЧ и молекула нуклеиновой кислоты включает нуклеотидные последовательности, кодирующие более одного регулярного белка ВИЧ. 13. The method of claim 5, wherein the pathogen is HIV and the nucleic acid molecule comprises nucleotide sequences encoding more than one regular HIV protein. 14. Способ по п. 5, отличающийся тем, что по крайней мере молекулы двух или более разных нуклеиновых кислот вводят в разные клетки индивидуума, причем разные нуклеиновые кислоты включают каждая нуклеотидные последовательности, кодирующие один или более антигенов патогена одного и того же патогена. 14. The method according to p. 5, characterized in that at least two or more different nucleic acid molecules are introduced into different cells of the individual, and different nucleic acids include each nucleotide sequence encoding one or more pathogen antigens of the same pathogen. 15. Способ по п. 5, отличающийся тем, что усилитель функции полинуклеотида и молекулу нуклеиновой кислоты вводят одновременно. 15. The method according to p. 5, characterized in that the polynucleotide function enhancer and the nucleic acid molecule are administered simultaneously. 16. Способ по п. 5, отличающийся тем, что индивидуумом является человек; усилитель функции полинуклеотида представлен бупивакаином; патоген представлен вирусом иммунодефицита человека; молекула нуклеиновой кислоты представлена ДНК и включает ДНК последовательность, кодирующую структурные белки gаg и ро1 ВИЧ с делецией psi, причем кодирующая gag и ро1 ДНК последовательность операбельно связана с усилителем вируса саркомы Paуса, непосредственно ранним промотором цитомегаловируса и малым сигналом полиаденилирования SV 40 и необязательно точкой начала репликации SV 40. 16. The method according to p. 5, characterized in that the individual is a person; an enhancer of polynucleotide function is represented by bupivacaine; the pathogen is a human immunodeficiency virus; the nucleic acid molecule is represented by DNA and includes a DNA sequence encoding the HIV gag and po1 structural proteins with a psi deletion; moreover, the gag and po1 DNA coding sequence is operably linked to a Paus sarcoma virus enhancer, directly an early cytomegalovirus promoter and a small SV 40 polyadenylation signal and optionally a start point SV 40 replication. 17. Способ по п. 16, отличающийся тем, что ДНК последовательность дополнительно включает rev-респондирующий элемент ВИЧ и делецию интегразы ВИЧ. 17. The method according to p. 16, wherein the DNA sequence further comprises a rev-responsive HIV element and a deletion of HIV integrase. 18. Способ по п. 17, отличающийся тем, что ДНК последовательность дополнительно включает сплайс-акцептор ВИЧ. 18. The method of claim 17, wherein the DNA sequence further comprises an HIV splice acceptor. 19. Способ по п. 16, отличающийся тем, что ДНК молекула дополнительно включает ДНК последовательность, кодирующую последовательность, дополнительно содержащую ВИЧ rev, операбельно связанный с промотором SV 40 и малым сигналом полиаденилирования SV 40 и необязательно с точкой начала репкации SV 40. 19. The method of claim 16, wherein the DNA molecule further comprises a DNA sequence encoding a sequence further comprising HIV rev, operably linked to the SV 40 promoter and the SV 40 small polyadenylation signal and optionally to the SV 40 replication start point. 20. Способ по п. 19, отличающийся тем, что кодирующая rev ДНК последовательность кодирует, кроме того, vpu ВИЧ и env ВИЧ. 20. The method according to p. 19, characterized in that the coding rev DNA sequence encodes, in addition, vpu HIV and env HIV. 21. Способ по п. 19, отличающийся тем, что ДНК последовательность, кодирующая gag и ро1, дополнительно включает rev-респондирующий элемент ВИЧ и делецию интегразы ВИЧ. 21. The method of claim 19, wherein the DNA sequence encoding gag and po1 further comprises a rev-responsive HIV element and a deletion of HIV integrase. 22. Способ по п. 21, отличающийся тем, что ДНК последовательность, кодирующая gag и ро1, дополнительно включает сплайс-акцептор ВИЧ. 22. The method according to p. 21, characterized in that the DNA sequence encoding gag and po1, further includes a splice acceptor of HIV. 23. Способ по п. 5, отличающийся тем, что индивидуумом является человек; усилитель функции полинуклеотида представлен бупиванаином; патоген представлен вирусом иммунодефицита человека; молекулами нуклеиновой кислоты является ДНК и содержит ДНК-последовательность, кодирующую rev-, vpu и env-белки HIV, операбельно связанную с усилителем вируса саркомы Рауса, непосредственно ранним промотором цитомегаловируса и малым сигналом полиаденилирования SV 40 и необязательно точкой начала репликации SV 40. 23. The method according to p. 5, characterized in that the individual is a person; an enhancer of polynucleotide function is represented by bupivanain; the pathogen is a human immunodeficiency virus; the nucleic acid molecule is DNA and contains a DNA sequence encoding HIV rev-, vpu and env proteins, operably linked to an Routh sarcoma virus enhancer, directly an early cytomegalovirus promoter, and a small polyadenylation signal SV40 and optionally SV40 replication origin. 24. Способ по п. 5, отличающийся тем, что индивидуумом является человек; усилитель функции полинуклеотида представлен бупивакаином; патоген представлен вирусом иммунодефицита человека; молекулы двух различных ДНК вводят в разные клетки индивидуума; молекула одной из нуклеиновых кислот является ДНК и включает ДНК последовательность, кодирующую структурные белки gag и ро1 ВИЧ с делецией psi, причем ДНК последовательность, кодирующая gag и pol, операбельно связана с усилителем вируса саркомы Рауса, непосредственно ранним промотором цитомегаловируса и малым сигналом полиаденилирования SV 40 и необязательно с точкой начала репликации SV 40; молекула другой нуклеиновой кислоты является ДНК и включает ДНК последовательность, кодирующую ВИЧ белки rev, vpu и env, операбельно связанные с усилителем вируса саркомы Рауса, непосредственно ранним промотором цитомегаловируса и малым сигналом полиаденилирования SV 40 и необязательно с точкой начала репликации SV 40. 24. The method according to p. 5, characterized in that the individual is a person; an enhancer of polynucleotide function is represented by bupivacaine; the pathogen is a human immunodeficiency virus; molecules of two different DNA are introduced into different cells of an individual; the molecule of one of the nucleic acids is DNA and includes a DNA sequence encoding the HIV gag and po1 structural proteins with a psi deletion, and the DNA sequence encoding gag and pol is operably linked to an Routh sarcoma virus enhancer, a direct early cytomegalovirus promoter and a small SV 40 polyadenylation signal and optionally with an SV 40 origin of replication; the other nucleic acid molecule is DNA and includes a DNA sequence encoding the HIV proteins rev, vpu, and env operably linked to an Routh sarcoma virus enhancer, directly an early cytomegalovirus promoter, and a small SV 40 polyadenylation signal and optionally with an SV 40 origin of replication. 25. Способ по п. 5, отличающийся тем, что осуществляют введение в клетки человека молекул двух разных нуклеиновых кислот, при это молекула каждой нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок, содержащий по меньшей мере один эпитоп, идентичный или по существу аналогичный эпитопу по меньшей мере одного антигена ВИЧ операбельно связанную с регуляторными последовательностями, причем нуклеотидные последовательности способны экспрессироваться в клетках, нуклеотидные последовательности каждой из двух разных нуклеиновых кислот кодируют различные белки, указанные белки имеют по меньшей мере один эпитоп, идентичный или по существу аналогичный эпитопу по меньшей мере одного белка ВИЧ, кодируемого генами ВИЧ, выбранными из группы, включающей gag, ро1 и env. 25. The method according to p. 5, characterized in that the introduction into human cells of molecules of two different nucleic acids, the molecule of each nucleic acid comprising a nucleotide sequence encoding a protein containing at least one epitope that is identical or essentially similar to the epitope according to at least one HIV antigen operably linked to regulatory sequences, the nucleotide sequences being able to be expressed in cells, the nucleotide sequences of each of two different nucleic acids encode different proteins, these proteins have at least an epitope identical or substantially similar to an epitope of at least one HIV protein encoded by HIV genes selected from the group consisting of gag, PO1 and env. 26. Способ по п. 1, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую целевой белок, содержащий эпитоп, идентичный или по существу подобный эпитопу белка, ассоциированного с клетками, характеризующими заболевание. 26. The method according to p. 1, characterized in that the nucleic acid molecule comprises a nucleotide sequence encoding a target protein containing an epitope that is identical or substantially similar to the epitope of a protein associated with cells characterizing the disease. 27. Способ по п. 26, отличающийся тем, что усилитель функции полинуклеотида представлен бупивакаином. 27. The method according to p. 26, wherein the polynucleotide function enhancer is represented by bupivacaine. 28. Способ по п. 26, отличающийся тем, что заболевание характеризуется гиперпролиферацией клеток. 28. The method according to p. 26, characterized in that the disease is characterized by hyperproliferation of cells. 29. Способ по п. 26, отличающийся тем, что заболевание является аутоиммунным заболеванием. 29. The method according to p. 26, wherein the disease is an autoimmune disease. 30. Способ по п. 26, отличающийся тем, что молекула нуклеиновой кислоты является молекулой ДНК. 30. The method according to p. 26, wherein the nucleic acid molecule is a DNA molecule. 31. Способ по п. 26, отличающийся тем, что молекулу нуклеиновой кислоты вводят внутримышечно. 31. The method according to p. 26, wherein the nucleic acid molecule is administered intramuscularly. 32. Способ по п. 26, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую целевой белок, выбранный из группы, включающей белковые продукты онкогенов myb, myc, fyn, ras, sarc, neu и trk; белковые продукты транслокационного гена bcl/ab1; Р53; EGRF; вариабельные области антител, полученных с помощью В-клеточных лимфом, и вариабельные области Т-клеточных рецепторов Т-клеточных лимфом. 32. The method according to p. 26, wherein the nucleic acid molecule comprises a nucleotide sequence encoding a target protein selected from the group consisting of protein products of the oncogenes myb, myc, fyn, ras, sarc, neu and trk; protein products of the bcl / ab1 translocation gene; P53; EGRF; the variable regions of antibodies obtained using B-cell lymphomas; and the variable regions of T-cell receptors of T-cell lymphomas. 33. Способ по п. 26, отличающийся тем, что белок выбирают из группы, включающей вариабельные области антител, участвующих в опосредуемом В-клетками аутоиммунном заболевании, и вариабельные области Т-клеточных рецепторов, участвующих в опосредуемом Т-клетками аутоиммунном заболевании. 33. The method according to p. 26, wherein the protein is selected from the group comprising variable regions of antibodies involved in B-cell-mediated autoimmune disease and variable regions of T-cell receptors involved in T-cell-mediated autoimmune disease. 34. Способ по п. 1, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок, присутствие которого будет компенсировать отсутствующий, нефункциональный или частично функционирующий белок или обеспечивать терапевтический эффект у индивидуума. 34. The method according to p. 1, characterized in that the nucleic acid molecule comprises a nucleotide sequence encoding a protein, the presence of which will compensate for the missing, non-functional or partially functioning protein or provide a therapeutic effect in an individual. 35. Способ по п. 34, отличающийся тем, что усилитель функции полинуклеотида представлен бупивакаином. 35. The method of claim 34, wherein the polynucleotide function enhancer is bupivacaine. 36. Способ по п. 34, отличающийся тем, что молекула нуклеиновой кислоты представлена молекулой ДНК. 36. The method according to p. 34, characterized in that the nucleic acid molecule is a DNA molecule. 37. Способ по п. 34, отличающийся тем, что молекула нуклеиновой кислоты вводится внутримышечно. 37. The method according to p. 34, wherein the nucleic acid molecule is administered intramuscularly. 38. Способ по п. 34, отличающийся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок, выбранный из группы, состоящей из ферментов, структурных белков, цитокинов, лимфокинов и факторов роста. 38. The method of claim 34, wherein the nucleic acid molecule comprises a nucleotide sequence encoding a protein selected from the group consisting of enzymes, structural proteins, cytokines, lymphokines and growth factors. 39. Фармацевтическая композиция, содержащая молекулу нуклеиновой кислоты, представленную молекулой ДНК, которая включает ДНК-последовательность, кодирующую белок, необязательно связанную с регуляторными элементами; усилитель функции полинуклеотида. 39. A pharmaceutical composition comprising a nucleic acid molecule represented by a DNA molecule, which includes a DNA sequence encoding a protein, optionally associated with regulatory elements; polynucleotide function enhancer. 40. Композиция по п. 39, отличающаяся тем, что молекула нуклеиновой кислоты включает нуклеотидную последовательность, кодирующую белок, выбранный из группы, состоящей из белков, включающих по крайней мере один эпитоп, идентичный или по существу подобный эпитопу антигена патогена; белков, содержащих эпитоп, идентичный или по существу подобный эпитопу белка, ассоциированного с гиперпролиферирующими клетками; белков, содержащих эпитоп, идентичный или по существу подобный эпитопу белка, ассоциированного с клетками, характеризующими аутоиммунное заболевание; белков, присутствие которых будет компенсировать отсутствующий нефункциональный или частично функционирующий белок у индивидуума; и белков, обеспечивающих терапевтический эффект у индивидуума. 40. The composition of claim 39, wherein the nucleic acid molecule comprises a nucleotide sequence encoding a protein selected from the group consisting of proteins comprising at least one epitope that is identical or substantially similar to an epitope of a pathogen antigen; proteins containing an epitope that is identical or substantially similar to an epitope of a protein associated with hyperproliferating cells; proteins containing an epitope identical or substantially similar to an epitope of a protein associated with cells characterizing an autoimmune disease; proteins whose presence will compensate for an absent non-functional or partially functioning protein in an individual; and proteins that provide a therapeutic effect in an individual. 41. Композиция по п. 39, отличающаяся тем, что патоген представляет собой вирус, выбранный из группы, состоящей из вируса иммунодефицита человека (HIV); вируса Т-клеточного лейкоза человека (HTLV); вируса гриппа; вируса гепатита A (HAV); вируса гепатита В (HBV); вируса гепатита С (HCV); вируса папилломы человека (HPV); вируса простого герпеса 1 (HSVI); вируса простого герпеса 2 (HSV2); цитомегаловируса (CMV); вируса Эпштейна-Барра (EBV); риновируса и коронавируса. 41. The composition according to p. 39, wherein the pathogen is a virus selected from the group consisting of human immunodeficiency virus (HIV); human T-cell leukemia virus (HTLV); influenza virus; hepatitis A virus (HAV); hepatitis B virus (HBV); hepatitis C virus (HCV); human papillomavirus (HPV); herpes simplex virus 1 (HSVI); herpes simplex virus 2 (HSV2); cytomegalovirus (CMV); Epstein-Barr virus (EBV); rhinovirus and coronavirus. 42. Композиция по п. 39, отличающаяся тем, что молекула нуклеиновой кислоты представляет собой ДНК-последовательность, кодирующую структурные белки gag и ро1 ВИЧ с делецией psi, причем последовательность ДНК, кодирующая gag и po1, операбельно связанную с усилителем вируса саркомы Рауса, непосредственно ранним промотором цитомегаловируса, малым сигналом полиаденилирования SV 40 и необязательно с точкой начала репликации SV 40. 42. The composition according to p. 39, wherein the nucleic acid molecule is a DNA sequence encoding the structural proteins gag and po1 of HIV with a deletion of psi, the DNA sequence encoding gag and po1 operably linked to an amplifier of the Routh sarcoma virus, directly an early cytomegalovirus promoter, a small SV 40 polyadenylation signal, and optionally with an SV 40 origin of replication. 43. Композиция по п. 39, отличающаяся тем, что молекула нуклеиновой кислоты представляет собой молекулу ДНК, включающую последовательность ДНК, кодирующую белки rev, vpu и env ВИЧ, операбельно связанную с усилителем вируса саркомы Рауса, непосредственно ранним промотором цитомегаловируса, малым сигналом полиаденилирования SV 40 и необязательно точкой начала репликации SV 40. 43. The composition according to p. 39, wherein the nucleic acid molecule is a DNA molecule comprising a DNA sequence encoding HIV rev, vpu and env proteins operably linked to an Routh sarcoma virus enhancer, directly an early cytomegalovirus promoter, a small polyadenylation signal SV 40 and optionally the start point of SV 40 replication. 44. Композиция по п. 39, отличающаяся тем, что усилитель функции полинуклеотида представлен бупивакаином. 44. The composition of claim 39, wherein the polynucleotide function enhancer is bupivacaine. 45. Фармацевтический набор для иммунизации, включающий первый инокулят, содержащий фармацевтически приемлемый носитель или разбавитель и молекулу первой нуклеиновой кислоты, включающую нуклеотидную последовательность, кодирующую по меньшей мере один белок ВИЧ, операбельно связанную с регуляторными последовательностями, причем нуклеотидная последовательность способна экспрессироваться в клетках человека, второй инокулят, содержащий фармацевтический приемлемый носитель или разбавитель и молекулу второй нуклеиновой кислоты, включающую нуклеотидную последовательность, кодирующую по меньшей мере один белок ВИЧ, операбельно связанную с регуляторными последовательностями, причем нуклеотидная последовательность способна экспрессироваться в клетках человека. при этом молекула первой нуклеиновой кислоты неидентична молекуле второй нуклеиновой кислоты. 45. A pharmaceutical immunization kit comprising a first inoculum comprising a pharmaceutically acceptable carrier or diluent and a first nucleic acid molecule comprising a nucleotide sequence encoding at least one HIV protein operably linked to regulatory sequences, the nucleotide sequence being able to be expressed in human cells, a second inoculum comprising a pharmaceutically acceptable carrier or diluent and a second nucleic acid molecule, including boiling nucleotide sequence encoding at least one HIV protein operably linked to regulatory sequences, wherein the nucleotide sequence is capable of being expressed in human cells. wherein the first nucleic acid molecule is not identical to the second nucleic acid molecule. 46. Набор по п. 45, отличающийся тем, что первый и второй инокуляты включают усилитель функции полинуклеотида или упомянутый фармацевтический набор дополнительно включает третий инокулят, содержащий усилитель функции полинуклеотида. 46. The kit of claim 45, wherein the first and second inoculums comprise a polynucleotide function enhancer or said pharmaceutical kit further comprises a third inoculum comprising a polynucleotide function enhancer. 47. Набор для иммунизации по п. 45, отличающийся тем, что молекула первой нуклеиновой кислоты и молекула второй нуклеиновой кислоты вместе кодируют белки gag, pol и env ВИЧ. 47. The immunization kit according to claim 45, wherein the first nucleic acid molecule and the second nucleic acid molecule together encode HIV gag, pol and env proteins. 48. Набор для иммунизации по п. 45, отличающийся тем, что включает первый инокулят, содержащий первую фармацевтическую композицию, включающую молекулу ДНК, которая содержит последовательность ДНК, кодирующую структурные белки gag и pol ВИЧ с делецией psi, причем упомянутая последовательность, кодирующая gag и pol, операбельно связана с усилителем вируса саркомы Pa- уса, непосредственно ранним промотором цитомегаловируса, малым сигналом полиаденилирования SV 40 и необязательно с точкой начала репликации SV 40; второй инокулят, содержащий вторую фармацевтическую композицию, включающую молекулу ДНК, которая содержит последовательность ДНК, кодирующую белки rev, vpu и env ВИЧ, операбельно связанную с усилителем вируса саркомы Рауса, непосредственно ранним промотором цитомегаловируса, малым сигналом полиаденилирования SV 40 и необязательно с точкой начала репликации SV 40. 48. The immunization kit according to claim 45, characterized in that it comprises a first inoculum containing a first pharmaceutical composition comprising a DNA molecule that contains a DNA sequence encoding the HIV gag and pol structural proteins with a psi deletion, said gag and pol is operably linked to an amplifier of the Pauss sarcoma virus, directly an early cytomegalovirus promoter, a small SV 40 polyadenylation signal, and optionally with an SV 40 origin of replication; a second inoculum containing a second pharmaceutical composition comprising a DNA molecule that contains a DNA sequence encoding HIV rev, vpu and env proteins operably linked to an Routh sarcoma virus enhancer, directly an early cytomegalovirus promoter, a small SV 40 polyadenylation signal, and optionally with a replication origin SV 40.
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