BACTERIOPHAGE-POLYMER HYBRID
The invention provides a targeted bacteriophage-polymer complex comprising a recombinant targeted-bacteriophage and a cationic polymer. The complex has a net positive charge. The invention provides methods of preparing bacteriophages and complexes thereof, and to their uses for the delivery of transgenes in a variety of gene therapy applications.
1 . A targeted bacteriophage-polymer complex comprising a recombinant targeted-bacteriophage and a cationic polymer, wherein the complex has a net positive charge.
2 . A complex according to claim 1 , wherein the recombinant bacteriophage comprises a nucleic acid sequence, which encodes a protein ligand that is capable of being expressed in the capsid coat, and which is also specific for a protein expressed on a target cell/tissue, so as to enable targeted delivery thereto, wherein the ligand is a tumour-targeting ligand, and wherein the ligand comprises the RGD4C ligand.
3 . (canceled)
4 . (canceled)
5 . A complex according to claim 1 , wherein the bacteriophage comprises a transgene, which exerts a therapeutic effect on a target cell, wherein the transgene comprises the Herpes simplex virus tyrosine kinase gene.
6 . (canceled)
7 . (canceled)
8 . A complex according to claim 1 , wherein the bacteriophage is F1, Fd or M13.
9 . (canceled)
10 . A complex according to claim 1 , wherein the cationic polymer is selected from a group consisting of: chitosan; poly-D-lysine (PDL); diethylaminoethyl (DEAE); diethylaminoethyl-dextran (DEAE.DEX); polyethyleneimine (PEI); polybrene; protamine sulphate; and a cationic lipid.
11 . A complex according to claim 10 , wherein the cationic lipid is selected from the group consisting of Fugene®, Lipofectamine®, and DOTAP (N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate).
12 . A complex according to claim 1 , wherein the cationic polymer comprises DEAE or DEAE.DEX.
13 . A complex according to claim 1 , wherein the polymer comprises PDL.
14 . A complex according to claim 1 , wherein the complex comprises a weight:weight ratio of about 50 ng-500 ng polymer:1 μg phage, or about 100 ng-400 ng polymer:1 μg phage, or about 130 ng-320 ng polymer:1 μg phage.
15 . A complex according to claim 1 , wherein the bacteriophage-cationic polymer complex comprises calcium phosphate.
16 . (canceled)
17 . (canceled)
18 . (canceled)
19 . A complex according to claim 1 , wherein the ζ-potential of the bacteriophage-polymer complex is at least 5 mV, 10 mV or 15 mV at physiological pH.
20 . A method of producing a targeted bacteriophage-polymer complex, the method comprising contacting a recombinant targeted-bacteriophage with a cationic polymer to form a stable complex, which has a net positive charge.
21 . (canceled)
22 . A method according to claim 20 , wherein the bacteriophage is incubated with the cationic polymer at a temperature of about 15-30° C., or about 18-25° C., or preferably about 20-23° C., for at least 5 mins, 10 mins or 15 mins.
23 . An aggregate comprising a plurality of bacteriophage-polymer complexes according to claim 1 .
24 . An aggregate according to claim 23 , wherein the mean particle size of the aggregate is between about 600 nm and 2500 nm, or between about 600 nm and 1500 nm, or between about 700 nm and 1200 nm.
25 . (canceled)
26 . (canceled)
27 . A vaccine comprising the bacteriophage-polymer complex according to claim 1 , and optionally an adjuvant.
28 . (canceled)
29 . (canceled)
30 . A genetic-molecular imaging technique comprising use of the bacteriophage-polymer complex according to claim 1 , optionally wherein the transgene encodes HSVtk and/or the sodium/iodide symporter (NIS).
31 . (canceled)
32 . A pharmaceutical composition comprising the bacteriophage-polymer complex according to claim 1 , and a pharmaceutically acceptable vehicle.
33 . (canceled)
34 . A method of treating, preventing or ameliorating a disease in a subject using a gene therapy technique, the method comprising administering, to a subject in need of such treatment, a therapeutically effective amount of the targeted bacteriophage-polymer complex according to the first aspect or the aggregate according to the third aspect.