US5166320A
Expired / Lapsed
US
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Carrier system and method for the introduction of genes into mammalian cells
Assignee: CONNECTICUT A CORP OF CT, University of
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Filing: 1990-04-02
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Granted: 1992-11-24
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Expires: 2010-04-02
Abstract
Abstract A targetable gene delivery system is provided for introducing foreign genes into mammalian cells. The system employs a soluble targetable DNA complex and utilizes receptor-mediated endocytosis to endow cell specificity. The soluble DNA-carrying complex is formed by non-covalently binding a ligand conjugate with the foreign gene. The conjugate, in turn, is formed by bonding receptor-specific ligands such as asialoglycoproteins to polycations such as polylysine through covalent bonds such as disulfide bonds. The system exhibits a high degree of cell specificity and offers potential for the treatment of inherited genetic disorders.
Inventors
- 👤 George Y. Wu
- 👤 Catherine H. Wu
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 03:42
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/62
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
73 words
A DNA carrier system soluble in a physiological fluid, comprising a molecular complex of: a) a conjugate comprising a ligand for an asialoglycoprotein receptor and a polycation which complexes the DNA and releases the DNA as a detectably active molecule within a cell; and b) DNA noncovalently complexed to the conjugate through the polycation, the complex having a proportion of conjugate to DNA wherein the said complex is soluble in a physiological fluid.
Claim 2
Dependent Claim
73 words
A DNA carrier system soluble in a physiological fluid, comprising a molecular complex of: a) a conjugate comprising a ligand for an asialoglycoprotein receptor and a polycation which complexes the DNA and releases the DNA as a detectably active molecule within a cell; and b) DNA noncovalently complexed to the conjugate through the polycation, the complex having a proportion of conjugate to DNA wherein the said complex is soluble in a physiological fluid.
Claim 3
Dependent Claim
13 words
A DNA carrier system of claim 1, wherein the ligand is an asialoglycoprotein.
Claim 4
Dependent Claim
12 words
A DNA carrier system of claim 1, wherein the polycation is polylysine.
Claim 5
Dependent Claim
20 words
A DNA carrier system of claim 1, wherein the DNA is a gene which can be expressed in the cell.
Claim 6
Dependent Claim
13 words
A DNA carrier system of claim 1, wherein the cell is an hepatocyte.
Claim 7
Dependent Claim
14 words
A polynucleotide carrier system of claim 1, wherein the ligand is a lactosylated protein.
Claim 8
Dependent Claim
19 words
A DNA carrier system of claim 1, wherein the ligand and the polycation are linked by a disulfide bond.
Claim 9
Independent Claim
69 words
A method of selectively introducing DNA into hepatocytes of a mammal, comprising administering to the mammal a molecular complex soluble in a physiological fluid comprising DNA linked to a conjugate comprising a ligand for an asialoglycoprotein receptor and a polycation which electrostatically complexes the DNA in amounts such that the complex is soluble and releases it within a cell to result in the internalization of a detectably active DNA.
Claim 10
Independent Claim
69 words
A method of selectively introducing DNA into hepatocytes of a mammal, comprising administering to the mammal a molecular complex soluble in a physiological fluid comprising DNA linked to a conjugate comprising a ligand for an asialoglycoprotein receptor and a polycation which electrostatically complexes the DNA in amounts such that the complex is soluble and releases it within a cell to result in the internalization of a detectably active DNA.
Claim 11
Dependent Claim
15 words
A method of claim 8, wherein the ligand is an asialoglycoprotein and polycation is polylysine.
Claim 12
Dependent Claim
12 words
A method of claim 9, wherein the DNA comprises an expressible gene.
Claim 13
Dependent Claim
77 words
A gene carrier system soluble in a physiological fluid and targetable to an asialoglycoprotein receptor-bearing cell, comprising a molecular complex of: a) a conjugate of a ligand specific for the asialoglycoprotein receptor linked to a polylysine; and b) a gene noncovalently complexed with the polylysine, the complex having a proportion of conjugate to gene wherein the said complex is soluble in a physiological fluid and wherein the gene is releasable as an expressible molecule in the cell.
Claim 14
Dependent Claim
77 words
A gene carrier system soluble in a physiological fluid and targetable to an asialoglycoprotein receptor-bearing cell, comprising a molecular complex of: a) a conjugate of a ligand specific for the asialoglycoprotein receptor linked to a polylysine; and b) a gene noncovalently complexed with the polylysine, the complex having a proportion of conjugate to gene wherein the said complex is soluble in a physiological fluid and wherein the gene is releasable as an expressible molecule in the cell.
Claim 15
Dependent Claim
17 words
A gene carrier system of claim 11, wherein the ligand is a glycoprotein containing terminal galactose residues.
Claim 16
Dependent Claim
17 words
A gene carrier system of claim 11, wherein the ligand is a protein having lactose coupled thereto.
Claim 17
Dependent Claim
22 words
A gene carrier system of claim 11, wherein the conjugate comprises a ratio of ligand to polylysine ranging from 4:1 to 7:1.
Claim 18
Dependent Claim
19 words
A gene carrier system of claim 14, wherein the ratio of conjugate to DNA ranges from 1:5 to 5:1.
Claim 19
Dependent Claim
18 words
A gene carrier system of claim 11, wherein the ligand is linked to the polylysine by disulfide bond.
Claim 20
Dependent Claim
15 words
A pharmaceutical composition comprising the gene carrier system of claim 11 in physiologically acceptable solution.
Claim 21
Independent Claim
65 words
A method of delivering a gene to hepatocytes, comprising injecting intravenously a pharmaceutical composition comprising a physiologically acceptable solution containing a molecular complex of a conjugate of a ligand specific for the asialoglycoprotein receptor linked to a polylysine and a gene noncovalently complexed with the polylysine, the complex having a proportion of conjugate to gene wherein the said complex is soluble in a physiological fluid.
Claim 22
Independent Claim
65 words
A method of delivering a gene to hepatocytes, comprising injecting intravenously a pharmaceutical composition comprising a physiologically acceptable solution containing a molecular complex of a conjugate of a ligand specific for the asialoglycoprotein receptor linked to a polylysine and a gene noncovalently complexed with the polylysine, the complex having a proportion of conjugate to gene wherein the said complex is soluble in a physiological fluid.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-11-24 | 1996-05-25 | 1996-11-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-11-24 | 2000-05-25 | 2000-11-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-11-24 | 2004-05-25 | 2004-11-24 | $7700.00 | Window Closed / Lapsed |