Methods for DNA-dependent targeting of a cell permeant antibody
AssigneeThe United States Government as represented by the Department of Veterans Affairs
InventorsRichard H. Weisbart, Robert N. Nishimura
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Abstract
The invention provides methods for selective targeting of live cells, which have undergone or are undergoing radiation or chemotherapy, at a site of interest with a cell-penetrating polypeptide. In one embodiment of the invention, the method comprises contacting the live cells with a cell-penetrating polypeptide comprising cell-penetrating determinants so that the cell-penetrating polypeptide binds extracellular DNA near or around the live cells so as to form a complex or association therewith such that the complex or associated polypeptide-DNA so bound bind the live cells and penetrates the live cells thereby selectively targeting live cells at a site of interest with a cell-penetrating polypeptide.
Claims40
A method for selectively targeting live cells at a site of interest with a cell-penetrating polypeptide, the method comprising:
- (a) introducing purified DNA near or around the live cells at the site of interest;
- (b) administering the cell-penetrating polypeptide comprising cell-penetrating determinants, before, after or concurrently with the purified DNA of step (a);
- (c) contacting the purified DNA or its degradation product with the cell-penetrating polypeptide so that the cell-penetrating polypeptide binds the purified DNA or its degradation product near or around the live cells so as to form a complex;
- (d) contacting one of the live cells with the complex in (c) so as to bind and penetrate the live cell;
- and
- (e) permitting additional complexes to form as in (c) and contacting additional cells with said complexes so as to bind and penetrate additional live cells at the site of interest;
- wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody,
- thereby, selectively targeting live cells at the site of interest with a cell-penetrating polypeptide.
A method for selective targeting of live cells at a site of interest, wherein the site of interest has undergone or is undergoing radiation or chemotherapy, with a cell-penetrating polypeptide, the method comprising:
- (a) administering purified DNA to the site of interest in an extracellular space;
- (b) contacting a live cell at the site of interest with a cell-penetrating polypeptide comprising cell-penetrating determinants so that the cell-penetrating polypeptide binds the purified DNA or its degradation product so as to form a complex;
- (c) contacting one of the live cells at the site of interest with the complex in (b) so as to bind and penetrate the live cell;
- (d) permitting additional complexes to form as in b and contacting additional cells with said complexes so as to bind and penetrate additional live cells at the site of interest;
- wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody, and
- thereby, selectively targeting live cells at the site of interest, which has undergone or is undergoing radiation or chemotherapy, with a cell-penetrating polypeptide.
A method for inhibiting cellular injury in a subject, the method comprising:
- (a) administering directly to the live cells at or near a site of cellular injury of the subject a cell-penetrating polypeptide comprising cell-penetrating determinants joined to a therapeutic agent;
- (b) contacting purified DNA with the cell-penetrating polypeptide so that the cell-penetrating polypeptide binds the purified DNA or its degradation product near or around the live cells so as to form a complex;
- (c) contacting one of the live cells with the complex in (b) so as to bind and penetrate the live cell;
- and
- (d) permitting additional complexes to form as in (b) and contacting additional cells with said complexes so as to bind and penetrate additional live cells at the site of cellular injury;
- wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody,
- thereby, inhibiting cellular injury in the subject.
A method for inducing cell death in a subject, the method comprising:
- (a) administering directly to the live cells at or near a site of injury of the subject a cell-penetrating polypeptide comprising cell-penetrating determinants joined to a therapeutic agent;
- (b) contacting purified DNA near or around the live cells with the cell-penetrating polypeptide so that the cell-penetrating polypeptide binds the purified DNA or its degradation product near or around the live cells so as to form a complex;
- (c) contacting one of the live cells with the complex in (b) so as to bind and penetrate the live cell which induces cell death;
- and
- (d) permitting additional complexes to form as in (b) and contacting additional cells with said complexes so as to bind and penetrate additional live cells inducing additional cell death at the site of injury;
- wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody,
- thereby inducing cell death at or near a site of injury with the cell-penetrating polypeptide.
A method for selective targeting of live cells at or near a site of cellular injury with a cell-penetrating polypeptide which comprises cell-penetrating determinants joined to a therapeutic agent, the method comprising:
- (a) contacting the live cells with a composition comprising
- (i) a cell-penetrating polypeptide which comprises cell-penetrating determinants joined to a therapeutic agent and
- (ii) purified DNA so that the cell-penetrating polypeptide binds the purified DNA or its degradation product near or around the live cells so as to form a complex such that the complex so formed binds one of the live cells and penetrates the live cell;
- and
- (b) permitting additional complexes to form as in (a) and contacting additional cells with said complexes so as to bind and penetrate additional live cells at the site of cellular injury,
- wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody,
- thereby selectively targeting live cells at a site of cellular injury with a cell-penetrating polypeptide.
A method for increasing uptake and enhancing efficacy of a cell-penetrating polypeptide comprising an anti-DNA antibody or a fragment in targeting tumor or cancer cells, the method comprising:
- (a) introducing purified DNA or artificial DNA at or near the tumor or cancer cells;
- (b) administering the cell-penetrating polypeptide;
- (c) allowing the cell-penetrating polypeptide to form additional complexes with the additional purified DNA or artificial DNA;
- and
- (d) permitting the additional complexes in (c) to contact the tumor or cancer cells,
- wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody,
- thereby increasing uptake and enhancing efficacy of a cell-penetrating polypeptide comprising an anti-DNA antibody or a fragment or a variant thereof in targeting tumor or cancer cells.
Dependencies are inferred from the claim text.