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US5225347A
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Therapeutic ribozyme compositions and expression vectors
Assignee: NEW YORK STATE SCIENCE & TECHNOLOGY FOUNDATION
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Filing: 1990-03-19
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Granted: 1993-07-06
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Expires: 2010-03-19
Abstract
Abstract Hepatitis delta is used as a vector for inhibition of viral infection and to express proteins in vivo in a cell-specific manner. The scope of delta's use as a vector is broadened in the present invention in several important ways. For example, a delta RNA genome capable of self-replication is enlarged to carry additional information, either coding for messenger RNA for a protein, or for a targeted ribozyme, which can be delivered to liver cells using delta's normally infectious properties, or to other cell types using chimeric delta viral agents carrying altered surface proteins. In another embodiment, the delta vector is made self-limiting, so that its role in delivering targeted information is separated from its viral property of unlimited infectious replication. Targeting is achieved through the use of sequences flanking the delta sequences that have affinity for sites on RNA to be cleaved.
Inventors
- 👤 Allan R. Goldberg
- 👤 Shaji T. George
- 👤 Hugh D. Robertson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 08:06
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C12N15/1131; C12N15/1132; A; A61; A61K; A61K47/00; A61K47/50; A61K47/69; A61K47/6901; A61P; A61P31/00; A61P31/12; C07; C07K; C07K14/00; C07K14/005; C12N15/63; C12N15/79; C12N15/85; C12N15/86; A61K38/00; A61K48/00; C07K2319/00; C12N2310/00; C12N2310/10; C12N2310/11; C12N2310/111; C12N2310/12; C12N2310/123; C12N2730/00; C12N2730/00011; C12N2730/10011; C12N2730/10111; C12N2730/10122; C12N2730/10134; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/16011; C12N2740/16041; C12N2740/16043; C12N2740/16111; C12N2740/16122; C12N2740/16211; C12N2740/16222; C12N2760/00; C12N2760/00011; C12N2760/10011; C12N2760/10111; C12N2760/10122; C12N2760/10134
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
94 words
A vector derived from hepatitis delta virus for delivering an RNA fragment to a cell comprising hepatitis delta virus RNA including fragments of RNA capable of replicating the vector in the cell, the vector having inserted therein the RNA fragment to be delivered, wherein the inserted RNA fragment to be delivered is inserted within the region of the hepatitis delta virus RNA not required for replication and so that it does not interrupt hepatitis delta virus RNA forming secondary and tertiary structure required for replication of the vector, including the regions with ribozyme activity.
Claim 2
Dependent Claim
94 words
A vector derived from hepatitis delta virus for delivering an RNA fragment to a cell comprising hepatitis delta virus RNA including fragments of RNA capable of replicating the vector in the cell, the vector having inserted therein the RNA fragment to be delivered, wherein the inserted RNA fragment to be delivered is inserted within the region of the hepatitis delta virus RNA not required for replication and so that it does not interrupt hepatitis delta virus RNA forming secondary and tertiary structure required for replication of the vector, including the regions with ribozyme activity.
Claim 3
Dependent Claim
71 words
The vector of claim 1 wherein the inserted RNA fragment to be delivered comprises, in addition to the regions of the hepatitis delta virus RNA required for replication of the vector, a sequential hepatitis delta virus RNA fragment selected from the group consisting of regions between residues 611 and 771 on the genomic strand and regions between residues 816 and 980 on the complementary anti-genomic strand, the fragment having ribozyme activity.
Claim 4
Dependent Claim
23 words
The vector of claim 1 further comprising RNA fragments targeting the inserted RNA fragment to be delivered to a defined target RNA molecule.
Claim 5
Dependent Claim
24 words
The vector of claim 1 wherein the inserted RNA fragment to be delivered comprises a hammerhead ribozyme of class derived from plant viroid pathogens.
Claim 6
Dependent Claim
15 words
The vector of claim 1 inserted into a cell line expressing hepatitis B surface antigen.
Claim 7
Dependent Claim
11 words
The vector of claim 5 wherein the cells are hepatic cells.
Claim 8
Dependent Claim
55 words
The vector of claim 1 wherein the inserted RNA fragment cleaves, or is transcribed into an RNA fragment that cleaves, a separate RNA molecule selected from the group consisting of oncogenes, tumor supporessor genes, viral genes, and cellular mRNAs which encode proteins selected from the group consisting of enzymes, hormones, cofactors, antibodies, and growth factors.
Claim 9
Dependent Claim
21 words
The vector of claim 1 encoding a chimeric protein including a non-hepatitis delta viral protein and a hepatitis delta viral protein.
Claim 10
Dependent Claim
17 words
The vector of claim 8 in a CD4+ cell line containing DNA encoding human immunodeficiency virus proteins.
Claim 11
Dependent Claim
88 words
A vector comprising a retrovirus incapable of producing infectious virions by itself, RNA fragments having ribozyme activity or transcribed into RNA fragments, the fragments cleaving a separate RNA molecule and RNA fragments targeting the RNA fragments having ribozyme activity, so that the RNA fragments with ribozyme activity will cleave protein-encoding viral RNA molecules and the RNA fragments having ribozyme activity are derived from hepatitis delta virus the molecules being essential for replication of the virus, wherein the targeted fragments with ribozyme activity cleave the target viral RNA molecules.
Claim 12
Dependent Claim
88 words
A vector comprising a retrovirus incapable of producing infectious virions by itself, RNA fragments having ribozyme activity or transcribed into RNA fragments, the fragments cleaving a separate RNA molecule and RNA fragments targeting the RNA fragments having ribozyme activity, so that the RNA fragments with ribozyme activity will cleave protein-encoding viral RNA molecules and the RNA fragments having ribozyme activity are derived from hepatitis delta virus the molecules being essential for replication of the virus, wherein the targeted fragments with ribozyme activity cleave the target viral RNA molecules.
Claim 13
Dependent Claim
16 words
The vector of claim 10 wherein the targeted molecules to be cleaved encode hepatitis B proteins.
Claim 14
Dependent Claim
17 words
The vector of claim 10 wherein the targeted molecules to be cleaved encode human immunodeficiency virus proteins.
Claim 15
Dependent Claim
33 words
The vector of claim 10 wherein the target RNA molecule encodes env protein of HIV and the replication of the defective retroviral vector is rendered self-limiting by the inhibition of env protein synthesis.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-07-05 | 1997-01-04 | 1997-07-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-07-05 | 2001-01-04 | 2001-07-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-07-05 | 2005-01-04 | 2005-07-06 | $7700.00 | Window Closed / Lapsed |