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WO1996004294A1 - Acide nucleique pour initier l'activite de la ribonuclease ou de la transcriptase inverse - Google Patents

Acide nucleique pour initier l'activite de la ribonuclease ou de la transcriptase inverse Download PDF

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Publication number
WO1996004294A1
WO1996004294A1 PCT/EP1995/003059 EP9503059W WO9604294A1 WO 1996004294 A1 WO1996004294 A1 WO 1996004294A1 EP 9503059 W EP9503059 W EP 9503059W WO 9604294 A1 WO9604294 A1 WO 9604294A1
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WO
WIPO (PCT)
Prior art keywords
nucleic acid
rna
trna
lys3
reverse transcriptase
Prior art date
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PCT/EP1995/003059
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German (de)
English (en)
Inventor
Michael STÜRZL
Matthias GÖTTE
Hermann Heumann
Wolfgang Brysch
Original Assignee
MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. filed Critical MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V.
Priority to EP95929041A priority Critical patent/EP0775153A1/fr
Publication of WO1996004294A1 publication Critical patent/WO1996004294A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1131—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against viruses
    • C12N15/1132—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against viruses against retroviridae, e.g. HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/353—Nature of the modification linked to the nucleic acid via an atom other than carbon
    • C12N2310/3531—Hydrogen

Definitions

  • the invention relates to a nucleic acid for the initiation of the activity of RNase H or reverse transcriptase, a method for producing such a nucleic acid and the use thereof.
  • AIDS acquired immunodeficiency syndrome
  • Retroviruses are therefore studied intensively. They have an RNA genome that is translated into a DNA molecule. As such, they integrate into the DNA of infected cells. The integrated DNA acts as a template for the synthesis of viral mRNA. RNA is translated into DNA by a retroviral enzyme called reverse transcriptase (RT). This enzyme requires a primer that binds to a specific location on the viral RNA, the primer binding site (PBS). One such primer is the cellular tRNA Lv ⁇ 3 . This is packaged (selected) in the viral capsid during the maturation of the virus particles. This ensures the initiation of the translation of RNA into DNA.
  • RT reverse transcriptase
  • the reverse transcriptase synthesizes a DNA strand complementary to the RNA on the primer and simultaneously degrades the transcribed RNA strand with its RNase H activity. A DNA double strand is then formed by the DNA-dependent polymerase activity of the reverse transcriptase.
  • the present invention is therefore based on the object of providing a means with which the replication of retroviruses can be inhibited.
  • the present invention is based on the idea of using the activity of an RNase H or a reverse transcriptase to inhibit retroviral replication.
  • RNase H is an enzyme of the cell as well as the virus, especially the retrovirus. In the latter case, RNase H is associated with reverse transcriptase.
  • the term "RNase H” used in the claims includes such an above enzyme.
  • An RNase H has the activity of degrading the RNA in an RNA / DNA hybrid.
  • RNA can be of any type and of any origin. It is preferably cellular or viral RNA, particularly preferably retroviral and very particularly preferably an RNA comprising the primer binding site or parts thereof. Cellular tRNA Lvs3 is also particularly preferred and that part of this tRNA which binds to the primer binding site is very particularly preferred.
  • the above DNA sequence is intended to degrade the complementary RNA through the activity of an RNase H. For example, the former is degraded with a DNA sequence that is complementary to the primer binding site.
  • a reverse transcriptase is a retroviral enzyme that translates RNA into DNA.
  • the reverse transcriptase requires the binding of an RNA primer to the primer binding site.
  • One such RNA primer is the cellular tRNA Lvs3 .
  • Recent investigations indicate that an RNA / RNA or RNA / DNA complex is essential for the initiation of the activity of a reverse transcriptase, the nucleotide sequence being of little importance.
  • RNA sequence which binds to a site different from the primer binding site.
  • the RNA sequence preferably comprises 18 nucleotides. Furthermore, a conserved site in the retroviral RNA is preferred as the site to be bound.
  • the RNA sequence is intended to redirect the natural initiation site of the reverse transcription to one or more artificial sites.
  • the RNA / DNA hybrids initiated there are subjected to the RNase H associated with the reverse transcriptase. An uncontrolled arises
  • the above nucleic acids preferably have structural elements which allow colocalization of the nucleic acids with the RNA to be bound, that is to say packaging both into the retroviral capsid.
  • Such structural elements can be cellular tRNA molecules that are selected specifically for the virus. HIV selects tRNA Lys ⁇ tRNA Lys2 , tRNA ,, e and tRNA Lvs3 . These tRNA molecules and parts thereof, for example the anticodon-stem-loop structure of the tRNA Lvs3 or a part thereof, are seen as preferred structural elements.
  • the nucleic acids which initiate the activity of an RNase are present as DNA analogs of the cellular tRNA Lys3 , ie as tDNA Lvs3 or modulated tDNA- Lvs3 .
  • the latter does not bind to the primer binding site, but, depending on the modulation, to other retroviral and / or cellular sequences.
  • nucleic acids which initiate the activity of a reverse transcriptase at artificial sites are particularly preferably present as modulated tRNA Lys3 analogs.
  • nucleic acids are preferred which are chimera from parts of tDNA Lys3 or modulated tDNA Lys3 and of tRNA Ly ⁇ 3 or modulated tRNA Lys3 .
  • the above nucleic acids can also have chemically modified deoxy or ribonucleotides. This increases the intracellular stability of the nucleic acids, in particular if the modifications are present at the 3 'end of the nucleic acids. Modifications of deoxy or ribonucleotides are known. As an example, reference is made to FIGS. 1 and 2. 1 shows the chemical structure of an oligodeoxyribonucleotide portion, FIG. 2 shows the chemical structure of an oligoribonucleotide portion. The structures shown in FIG. 1 and FIG. 2 are understood as sections from longer oligonucleotide chains.
  • the letter B denotes an organic base, such as adenosine (A), guanosine (G), cytidine (C), thymidine (T) or uridine (U), which is attached to the N9 (A, G) or N1 ( C, T, U) position is coupled to the deoxyribose.
  • the oligodeoxy ribonucleotide portions can e.g. be chemically modified as follows:
  • the letter B denotes an organic base such as adenosine (A), guanosine (G), cytidine (C) or uridine (U), which is at the N9 (A, G) or N1 (C, U) position the ribose is coupled.
  • the oligoribonucleotide components can be chemically modified, for example, as follows: 4. All R 1 positions of an oligoribonucleotide portion are substituted
  • R 1 positions of an oligoribonucleotide portion are alternately substituted 5 ' Bp- (BpBp) n -B- p -B
  • the nucleic acid fraction of the nucleic acids described in the present invention can consist of a fraction as described under 1.1-1.6, 2.1-2.6, 3.1-3.6, 4.1-4.6, 5.1-5.6, 6.1-6.6 or 7.1-7.3, whose base sequence is complementary to a partial sequence of the viral or cellular target RNA.
  • one or more nucleic acid segments can be found, as under 1.1-1.6, 2.1-2.6, 3.1-3.6, 4.1-4.6, 5.1-5.6, 6.1-6.6, 7.1-7.3 described, which, due to their respective base sequence, form a secondary and tertiary structure which specifically bind to viral proteins, for example reverse transcriptase, nucleocapsid
  • nucleic acid segments can also contain base modifications, as shown for example in FIG. 3.
  • nucleic acids described in the present invention can also be other molecules, e.g. Contain coupled peptides, proteins, phospholipids, steroids, which specifically bind to viral or cellular proteins instead of or in addition to a nucleic acid secondary or tertiary structure.
  • the nucleic acids are preferably with a substance which inhibits virus multiplication, particularly preferably ribozyme or 2 ',
  • a method for producing the above nucleic acids is also provided.
  • Such a process advantageously comprises the following process steps:
  • step (f) Carrying out a further synthesis cycle or cleaving the nucleotide chain from the carrier.
  • the above chemical modifications are inserted in step (d). This can be done in the usual way.
  • Nucleic acids according to the invention can be introduced into a person to be treated by conventional methods.
  • T-lymphocytes can be isolated from an HIV-infected person and the nucleic acids according to the invention can be introduced into them by known measures, such as electroporation.
  • nucleic acids according to the invention are also possible to produce, ie to express, nucleic acids according to the invention, especially if they consist of ribonucleotides, such as modulated tRNA Lys3 (see FIG. 4), only in the person to be treated.
  • nucleic acids according to the invention it is advantageous to insert the DNA coding for such a nucleic acid into an expression vector such as pNEOtp-muNTSI (cf. Hemann et al., DNA and Cell Biology, (1994) and the expression plasmid obtained, as indicated above, in It goes without saying that the above DNA can comprise all sequences which can influence the expression of a nucleic acid according to the invention.
  • FIG. 5 in which a DNA is indicated which is suitable for tRNA Ly " 3 (a) or modulated tRNAL Ly83 (b)
  • the bold sequence relates to (modulated) tRNA Ly ⁇ 3
  • the weakly printed sequence relates to regulatory regions such as the termination sequence of polymerase III (oligo No. 9).
  • the DNA of FIG. 5 is inserted into the expression vector pNEOtp-muNTSI digested with Sall, whereby an expression plasmid suitable for introduction into a person to be treated is obtained will.
  • Nucleic acids according to the invention are distinguished in that they are targeted
  • RNA particularly cellular or viral, very particularly retroviral RNA.
  • the activity of cellular as well as viral RNase H, in particular associated with reverse transcriptase is used.
  • Nucleic acids according to the invention are therefore ideally suited for inhibiting the replication of retroviruses, in particular of HIV viruses. The present invention thus opens up the possibility of diagnosing and treating infections or diseases which are caused by the above viruses.
  • FIG. 2 shows the chemical structure of an oligoribonucleotide portion of a nucleic acid according to the invention
  • FIG. 3 shows the structure of tRNA Ly83 and modified bases thereof
  • Fig. 4 shows the structure of tRNA Ly " 3 and modulated tRNA Ly * 3
  • Fig. 5 shows the DNA for tRNA Ly, 3 (a) and modulated tRNA Ly, 3 (b)
  • FIG. 6 shows chemically treated tDNA Ly ⁇ 3 under native and denaturing conditions
  • FIG. 7 shows the titration of the tDNA Ly ⁇ 3 and tRNA Lys3 with HIV-1-RT
  • Figure 8 shows the RNA region of HIV-1 and the primer binding site
  • tDNA Lva3 The DNA analogue of tRNA Lys3 (see FIG. 3) was as described above ben, chemically synthesized. An automatic synthesizer from Applied Biosystems was used for this. After the synthesis, the tDNA Lys3 was purified by gel electrophoresis. The sequence of the tDNA Lys3 is identical to that of the tRNA Lys3 . However, it has no modified bases. The uridine ribonucleosides are also replaced by corresponding thymidine deoxynucleotides.
  • the tDNA ys3 was radioactively marked on the 3'termin or alternatively 5'terminus.
  • the 3'terminal labeling was carried out enzymatically using "transferase” and [ ⁇ - 32 P] dd ATP.
  • the 5 'labeling was also carried out enzymatically using polynucleotide kinase and [- 32 P] ATP.
  • lane (1) of the OsO 4 and lane (3) of the DEPC reaction show the tDNA Lys3 under native conditions and lanes (2) and (4) show the tDNA Ly83 under denaturing conditions.
  • a comparison of the reactions between native and denaturing agents shows that thymidines and adenosines of tDNA Lys3 exist which are not modified and are therefore paired with bases.
  • the tDNA Lys3 structure determined in this way is a tRNA Lys3 structure analogous.
  • Example 2 Comparison of the interaction of the natural replication primer tRNA ' Ly * 3 and chemically synthesized tDNA Ly * 3 with the reverse transcriptase (RT) of HIV-1
  • tRNA Lys3 purified tRNA Lys3 isolated from rabbit liver was used (Raba et al., (1979) Eur.J.Biochem. 97, 305-318). This tRNA Lys3 is identical in sequence to the human tRNA Lys3 .
  • the 5 'end of the natural tRNA Lys3 was firstly dephosphorylated with alkaline phosphatase and then, as described above in Example 1, with l - 32 P] ATP radiolabeled.
  • Lane C shows the gel electrophoretic analysis of the two band-shift experiments.
  • Lane C is the control lane without HIV-1 RT
  • the other lanes show the titration of the nucleic acid with HIV-1 RT in the molar ratios 1: 1, 1: 2, 1: 3, 1: 4, 1: 5, 1: 6, 1: 7 and 1: 8.
  • the positions of the tRNA / RT or tDNA / RT complexes and the uncomplexed nucleic acids are marked in the figure.
  • RNA template was synthesized enzymatically by run-off transcription using the T7 RNA polymerase.
  • the RNA sequence is shown in Fig. 8.
  • the radioactive labeling at the 5 'terminus of the RNA template was carried out as described in Example 1 above.
  • a 3 'terminal labeling was carried out with [ 32 P] pCp and T4-RNA polymerase.
  • the primer and template were prehybridized in a molar ratio of 1.2: 1.
  • the components were first heat denatured, then 10 min. incubated at 55 ° C and finally cooled to 37 ° C.
  • RNA / RNA substrate remains intact, while the RNA / DNA substrate is hydrolyzed by the RT-associated RNaseH activity.
  • Example 4 Catalytic hydrolysis of the primer binding site by HIV-1-RT, nucleocapsid protein (NCp7) and tDNA Ly * 3 under native conditions
  • NCp7 was used (De Rocquigny et al., (1992) Proc.Natl.Acad, Sci. USA 89, 6472-6476).
  • RNA template 55 pmol RNA template were incubated with 5.5 pmol tDNA Ly ⁇ 3 and 15 pmol RT at 37 ° C. In separate reaction batches, 20 pmol of NCp7 was additionally incubated. The reactions were stopped after various times and analyzed by gel electrophoresis.
  • FIG. 9 shows that in the presence of NCp7 the 10-fold molar excess of RNA is completely hydrolyzed. The RNA hydrolysis is accelerated 20-fold compared to the control reaction without NCp7. It also shows that HIV-I-RT, NCp7 and tDNA Lys3 are the components of a system for the targeted catalytic degradation of the primer binding site. Since the above reaction conditions correspond to native conditions, the result obtained is of the greatest importance for the in vivo application of the present invention.
  • RNA template is characterized by the natural structure of the viral RNA genome.
  • the PBS is degraded in the presence of NCp7 or alternatively by prehydbridization.
  • structured RNA and DNA molecules are unfolded by NCp7, specifically hybridized and used as a substrate of the RNase H associated with RT.
  • the natural replication primer tRNA Lys3 packaged in the virus capsid can also be the target of DNA-induced hydrolysis.
  • the tRNA Lys3 acts as an RNA template for the RT / RNase H-catalyzed RNA
  • a ribozyme can also be used for tRNA Lys3 hydrolysis.
  • the colocalization of the rection partners in the capsid means that only the selected tRNA " Lys3 and not the cytoplasmic tRNA is hydrolyzed.

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Abstract

L'invention concerne un acide nucléique comportant une séquence liant un ARN, où la séquence (a) initie l'activité d'une ribonucléase H et est une séquence d'ADN, et/ou (b) initie l'activité d'une transcriptase inverse en un site différent du site de liaison d'amorce et est une séquence d'ARN. L'invention concerne en outre un procédé permettant de préparer un acide nucléique de ce type et son utilisation.
PCT/EP1995/003059 1994-08-01 1995-07-31 Acide nucleique pour initier l'activite de la ribonuclease ou de la transcriptase inverse WO1996004294A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP95929041A EP0775153A1 (fr) 1994-08-01 1995-07-31 Acide nucleique pour initier l'activite de la ribonuclease ou de la transcriptase inverse

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19944427219 DE4427219A1 (de) 1994-08-01 1994-08-01 Nukleinsäure zur Initiation der Aktivität von RNase H bzw. reverser Transkriptase
DEP4427219.7 1994-08-01

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WO1996004294A1 true WO1996004294A1 (fr) 1996-02-15

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997044064A3 (fr) * 1996-05-20 1998-05-14 Us Health Oligonucleotides qui lient specifiquement des proteines nucleocapsidiques retrovirales
US6776986B1 (en) 1996-06-06 2004-08-17 Novartis Ag Inhibition of HIV-1 replication by antisense RNA expression

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989008146A1 (fr) * 1988-02-26 1989-09-08 Worcester Foundation For Experimental Biology Inhibition du htlv-iii par des oligonucleotides exogenes
EP0339842A2 (fr) * 1988-04-27 1989-11-02 Isis Pharmaceuticals, Inc. Dérivés d'oligoribonucléotides et leur utilisation comme agents antiviraux
WO1991004753A1 (fr) * 1989-10-02 1991-04-18 Cetus Corporation Conjugues d'oligonucleotides non codants et leurs emplois therapeutiques
EP0463712A2 (fr) * 1990-06-27 1992-01-02 University Patents, Inc. Phosphorothioates des polynucléotides comme agents actifs contre les infections retrovirales
WO1993024133A1 (fr) * 1992-05-27 1993-12-09 City Of Hope MOLECULES CHIMERIQUES DE RIBOZYMES D'ARNt?LYS¿

Family Cites Families (1)

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Publication number Priority date Publication date Assignee Title
US4806463A (en) * 1986-05-23 1989-02-21 Worcester Foundation For Experimental Biology Inhibition of HTLV-III by exogenous oligonucleotides

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989008146A1 (fr) * 1988-02-26 1989-09-08 Worcester Foundation For Experimental Biology Inhibition du htlv-iii par des oligonucleotides exogenes
EP0339842A2 (fr) * 1988-04-27 1989-11-02 Isis Pharmaceuticals, Inc. Dérivés d'oligoribonucléotides et leur utilisation comme agents antiviraux
WO1991004753A1 (fr) * 1989-10-02 1991-04-18 Cetus Corporation Conjugues d'oligonucleotides non codants et leurs emplois therapeutiques
EP0463712A2 (fr) * 1990-06-27 1992-01-02 University Patents, Inc. Phosphorothioates des polynucléotides comme agents actifs contre les infections retrovirales
WO1993024133A1 (fr) * 1992-05-27 1993-12-09 City Of Hope MOLECULES CHIMERIQUES DE RIBOZYMES D'ARNt?LYS¿

Non-Patent Citations (8)

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Title
A.S. KHAN, B.A. ROE: "Aminoacylation of Synthetic DNAs Corresponding to E. Coli Phenylalanine and Lysine tRNAs", SCIENCE, vol. 241, pages 74 - 9 *
D. HERSCHLAG ET AL.: "Protein Enhancement of Hammerhead Ribozyme Catalysis", SCIENCE, vol. 262, pages 99 - 102 *
J. GOODCHILD: "Inhibition of Gene Expression by Oligonucleotides' in 'Oligodeoxynucleotides - Antisense Inhibitors of Gene Expression", J.S. COHEN (EDITOR) *
J.L. DARLIX ET AL.: "Interactions between HIV-1 nucleocapsid protein and viral DNA may have important functions in the viral life cycle", NUCLEIC ACIDS RES., vol. 21, pages 831 - 9 *
L. TARRAGO-LITVAK ET AL.: "Interaction of tRNALys with the p66/p66 Form of HIV-1 Reverse Transcriptase Stimulates DNA Polymerase and Ribonuclease H Activities", J. BIOL. CHEM., vol. 267, pages 19356 - 19362 *
R.S. GOODY ET AL.: "Synthetic human tRNALys-UUU and natutal bovine tRNALys-SUU interact with HIV-1 reverse transcriptase and serve as specific primers for retroviral cDNA synthesis", GENE, vol. 111, pages 183 - 197 *
S. LITVAK ET AL.: "Preferential Interaction of Human Immunodeficiency Virus Reverse Transcriptase with two Regions of Primer tRNALys as evidenced by Footprinting Studies and Inhibition with Synthetic Oligoribonucleotides", J. MOL. BIOL., vol. 226, pages 1 - 6 *
Z. TSUCHIHASHI, P.O. BROWN: "DNA Strand Exchange and Selective DNA annealing Promoted by the Human Immunodeficiency Virus Typ 1 Nucleocapsid Protein", J. VIROL., vol. 68, no. 9, pages 5863 - 70 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997044064A3 (fr) * 1996-05-20 1998-05-14 Us Health Oligonucleotides qui lient specifiquement des proteines nucleocapsidiques retrovirales
US6316190B1 (en) 1996-05-20 2001-11-13 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Oligonucleotides which specifically bind retroviral nucleocapsid proteins
US6776986B1 (en) 1996-06-06 2004-08-17 Novartis Ag Inhibition of HIV-1 replication by antisense RNA expression

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DE4427219A1 (de) 1996-02-08
EP0775153A1 (fr) 1997-05-28

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