CA2086577A1 - Potential target for immunotherapy to prevent replication of hiv (aids virus) in humans: cdna and protein for a putative regulator of hiv gag polyprotein expression reconstructed from the antisense strand of human p53 gene - Google Patents
Potential target for immunotherapy to prevent replication of hiv (aids virus) in humans: cdna and protein for a putative regulator of hiv gag polyprotein expression reconstructed from the antisense strand of human p53 geneInfo
- Publication number
- CA2086577A1 CA2086577A1 CA002086577A CA2086577A CA2086577A1 CA 2086577 A1 CA2086577 A1 CA 2086577A1 CA 002086577 A CA002086577 A CA 002086577A CA 2086577 A CA2086577 A CA 2086577A CA 2086577 A1 CA2086577 A1 CA 2086577A1
- Authority
- CA
- Canada
- Prior art keywords
- hiv
- protein
- cdna
- human
- gene
- Prior art date
- Legal status (The legal status 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 status listed.)
- Abandoned
Links
Classifications
-
- 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
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/82—Translation products from oncogenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Plant Pathology (AREA)
- Oncology (AREA)
- Physics & Mathematics (AREA)
- Toxicology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
ABSTRACT
A cDNA reconstructed from the antisense strand of human p53 gene encodes a 53 aa nuclear protein (Higaa) which has the potential for interacting with a domain of very highly conserved sequences in HIV gag polyprotein precursor. The protein which appears to be an intracellular activator of gag may be important for the successful replication of HIV in human cells. Since the protein is constitutively repressed by the p53 gene in normal cells, it is potentially a good target for a simple immunological approach to prevent multiplication of HIV following integration of the viral genome into the human genome.
A cDNA reconstructed from the antisense strand of human p53 gene encodes a 53 aa nuclear protein (Higaa) which has the potential for interacting with a domain of very highly conserved sequences in HIV gag polyprotein precursor. The protein which appears to be an intracellular activator of gag may be important for the successful replication of HIV in human cells. Since the protein is constitutively repressed by the p53 gene in normal cells, it is potentially a good target for a simple immunological approach to prevent multiplication of HIV following integration of the viral genome into the human genome.
Description
~ ~08~77 SPECIFICATION
HIV the AIDS causing virus will probably infect more than 100 million humans before the beginning of the 21 century, and although there is a very slight chance that a vaccine may be developed to prevent infection before the 21 century there i9 no hope of curing an infected person. Certain drugs based on nucleotide analogues, e.g., AZT, directed against HIV reverse transcriptase gene, have had qualified success in slowing the rate of intercellular spreading of the virus in small number of .
cases; however, the side effects of these drugs are almost unacceptable in most cases, and the cost of treatment is enormous. Replication of the virus, following integration into ;;
the human genome is a key step in the spread of infectivity. If replication could be blocked or slowed down then the . :-: - ~.:
intercellular infection will be considerably curtailed. The HIV
virus `gag' and `gagpol' genes are involved in crucial steps of viral replication; therefore, any interference with the expression or processing of `gag' will have a negative effect on viral replication. This invention relates to the discovery of a protein HIV gag activator protein (Higaa) encoded by a gene, antisense to a region of the human p53 gene, which may be required for the correct expression or processing of HIV gag polyprotein. p53 ls involved in protecting humans against invading virus especially tumour causing viruses.
~6577 (2) Because `Higaa' mRNA is antisense to the sense strand of p53 mRNA it is probably not expressed in normal humans, but may be expressed in humans suffering from certain virus, or other infections, which could hinder, or stop, antisense repression by p53 gene and hence allow `Higaa' to be expressed in sufficient quantities to activate HIV gag.
A region of the antisense strand of p53 cDNA comprising nucleotides 532 - 693 inclusive, is 100 % homologous to `Higaa' cDNA. The protein deduced from the cDNA is made up of 53 amino acids (53 aa) and by similarity appears to be a nuclear protein~
The structure of the cDNA is shown in the embodiment of figure la and that of the deduced protein in the embodiment of figure lb. The domain of the protein (16 aa) which is capable of interacting with gag polyprotein is overlined.
`Higaa' cDNA is recon~tructed (and Hind III restriction enzyme sites added) by oligonucleotide synthesis on an Applied Biosystems DNA synthesizer using the column method according to the recommendations of the manufacturer. The positive and negative strands are synthesised separately, gel purified on agarose, then mixed together in equimolar amounts and repurified by agarose gel electrophoresis. The cDNA construct is ligated into the pTrcHis Xpress vector and processed in the expresst system (Invitrogen corporation pTrcHis Xpress Prokaryotic Expression and Purification System) according to the manufacturers instructions.
~i 2~ 77 : ~
HIV the AIDS causing virus will probably infect more than 100 million humans before the beginning of the 21 century, and although there is a very slight chance that a vaccine may be developed to prevent infection before the 21 century there i9 no hope of curing an infected person. Certain drugs based on nucleotide analogues, e.g., AZT, directed against HIV reverse transcriptase gene, have had qualified success in slowing the rate of intercellular spreading of the virus in small number of .
cases; however, the side effects of these drugs are almost unacceptable in most cases, and the cost of treatment is enormous. Replication of the virus, following integration into ;;
the human genome is a key step in the spread of infectivity. If replication could be blocked or slowed down then the . :-: - ~.:
intercellular infection will be considerably curtailed. The HIV
virus `gag' and `gagpol' genes are involved in crucial steps of viral replication; therefore, any interference with the expression or processing of `gag' will have a negative effect on viral replication. This invention relates to the discovery of a protein HIV gag activator protein (Higaa) encoded by a gene, antisense to a region of the human p53 gene, which may be required for the correct expression or processing of HIV gag polyprotein. p53 ls involved in protecting humans against invading virus especially tumour causing viruses.
~6577 (2) Because `Higaa' mRNA is antisense to the sense strand of p53 mRNA it is probably not expressed in normal humans, but may be expressed in humans suffering from certain virus, or other infections, which could hinder, or stop, antisense repression by p53 gene and hence allow `Higaa' to be expressed in sufficient quantities to activate HIV gag.
A region of the antisense strand of p53 cDNA comprising nucleotides 532 - 693 inclusive, is 100 % homologous to `Higaa' cDNA. The protein deduced from the cDNA is made up of 53 amino acids (53 aa) and by similarity appears to be a nuclear protein~
The structure of the cDNA is shown in the embodiment of figure la and that of the deduced protein in the embodiment of figure lb. The domain of the protein (16 aa) which is capable of interacting with gag polyprotein is overlined.
`Higaa' cDNA is recon~tructed (and Hind III restriction enzyme sites added) by oligonucleotide synthesis on an Applied Biosystems DNA synthesizer using the column method according to the recommendations of the manufacturer. The positive and negative strands are synthesised separately, gel purified on agarose, then mixed together in equimolar amounts and repurified by agarose gel electrophoresis. The cDNA construct is ligated into the pTrcHis Xpress vector and processed in the expresst system (Invitrogen corporation pTrcHis Xpress Prokaryotic Expression and Purification System) according to the manufacturers instructions.
~i 2~ 77 : ~
(3) The 16 aa domain, overlined in the embodiment of figure lb, is probably all that is required to make antibodies, or very specific anti-peptide reagents, for blocking reaction of `Higaa' with HIV gag. Since the protein is not expressed in humans and does not have homology to any known, normal human, protein, blocking it should not produce undesirable side effects in AIDS
victims, and hence, a simple immunological approach to stop the intercellular spread of HIV infection which leads to full blown AIDS might be possible.
:
. :
victims, and hence, a simple immunological approach to stop the intercellular spread of HIV infection which leads to full blown AIDS might be possible.
:
. :
Claims (4)
1. The cDNA and protein sequences described in the embodiment of figure 1a and 1b.
2. Any diagnostic and or therapeutic reagents including ribozymes produced using the cDNA described in the embodiment of figure 1a or from any modified form of this cDNA.
3. Any diagnostic and or therapeutic reagents including polyclonal antibodies, monoclonal antibodies, fusion proteins, anti-peptide reagents etc., made from any part or all of amino acid sequence described in the embodiment of figure 1b.
4. Any use of the cDNA and or protein described in the embodiment of figure 1a and 1b or modifications of the cDNA or protein to treat any type of cancer.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002086577A CA2086577A1 (en) | 1992-12-31 | 1992-12-31 | Potential target for immunotherapy to prevent replication of hiv (aids virus) in humans: cdna and protein for a putative regulator of hiv gag polyprotein expression reconstructed from the antisense strand of human p53 gene |
| AU59690/94A AU5969094A (en) | 1992-12-31 | 1993-12-30 | Agents for the prevention and treatment of hiv replication and aids in humans |
| PCT/EP1993/003721 WO1994016066A1 (en) | 1992-12-31 | 1993-12-30 | Agents for the prevention and treatment of hiv replication and aids in humans |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002086577A CA2086577A1 (en) | 1992-12-31 | 1992-12-31 | Potential target for immunotherapy to prevent replication of hiv (aids virus) in humans: cdna and protein for a putative regulator of hiv gag polyprotein expression reconstructed from the antisense strand of human p53 gene |
| US11337093A | 1993-08-30 | 1993-08-30 | |
| PCT/EP1993/003721 WO1994016066A1 (en) | 1992-12-31 | 1993-12-30 | Agents for the prevention and treatment of hiv replication and aids in humans |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2086577A1 true CA2086577A1 (en) | 1994-07-01 |
Family
ID=25675789
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002086577A Abandoned CA2086577A1 (en) | 1992-12-31 | 1992-12-31 | Potential target for immunotherapy to prevent replication of hiv (aids virus) in humans: cdna and protein for a putative regulator of hiv gag polyprotein expression reconstructed from the antisense strand of human p53 gene |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU5969094A (en) |
| CA (1) | CA2086577A1 (en) |
| WO (1) | WO1994016066A1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6776986B1 (en) | 1996-06-06 | 2004-08-17 | Novartis Ag | Inhibition of HIV-1 replication by antisense RNA expression |
| US8575121B2 (en) * | 2007-11-12 | 2013-11-05 | The United States of America as represented by the Secetary of the Department of Health and Human Services | Therapeutic applications of p53 isoforms in regenerative medicine, aging and cancer |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NO312681B1 (en) * | 1990-08-24 | 2002-06-17 | Univ California | Process for the preparation of a pharmaceutical composition with suppressive action / activity |
-
1992
- 1992-12-31 CA CA002086577A patent/CA2086577A1/en not_active Abandoned
-
1993
- 1993-12-30 AU AU59690/94A patent/AU5969094A/en not_active Abandoned
- 1993-12-30 WO PCT/EP1993/003721 patent/WO1994016066A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| AU5969094A (en) | 1994-08-15 |
| WO1994016066A1 (en) | 1994-07-21 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued |