Targeted drug-carrying bacteriophages

Abstract

The present invention relates to the field of drug delivery. More specifically, the invention relates to the preparation and use of a bacteriophage conjugated through a labile/non labile linker or directly to at least 1,000 therapeutic drug molecules such that the drug molecules are conjugated to the outer surface of the bacteriophage. The bacteriophage optionally displays on its coat a ligand that endows it with specificity towards target cells. Thus, there is provided a targeted, high-capacity drug delivery system useful for the treatment of various pathological conditions.

1 . A pharmaceutical composition comprising bacteriophage-drug conjugates, wherein an average of at least 1,000 drug or isotope molecules is conjugated to an outer surface of each bacteriophage. 
     
     
         2 . The composition of  claim 1 , wherein the bacteriophage displays an exogenous targeting moiety which binds a cell surface molecule on a target cell. 
     
     
         3 . The composition of  claim 2 , wherein the targeting moiety is selected from the group consisting of an antibody, an antibody fragment, a peptide, a polypeptide, a carbohydrate, a lipid, a glycolipid, a nucleic acid and derivatives thereof. 
     
     
         4 . The composition of  claim 2 , wherein the targeting moiety comprises a ligand selected from: the ZZ domain derived from  Staphylococcus aureus  protein A, avidin, streptavidin, biotin and derivatives thereof, the ligand bound to a targeting molecule selected from: an antibody, an antibody fragment, a peptide, a polypeptide, a carbohydrate, a lipid, a glycolipid, a nucleic acid and analogs and derivatives thereof. 
     
     
         5 . The composition of  claim 2 , wherein the target cell is selected from the group consisting of a bacterial cell, a fungal cell, a yeast cell, a unicellular parasite cell, a multicellular parasite cell and a mammalian cell. 
     
     
         6 . The composition of  claim 1 , wherein the drug is selected from a group consisting of: an antibacterial agent, an antibiotic, an anti fungal drug, an anti viral drug, a parasiticide, a cytotoxic agent, a cytostatic agent, and a radioactive isotope. 
     
     
         7 . The composition of  claim 1 , wherein the drug is linked to a coat protein of the bacteriophage via a linker. 
     
     
         8 . The composition of  claim 7 , wherein the linker is a labile linker which upon cleavage releases the drug. 
     
     
         9 . The composition of  claim 8 , wherein the drug is in the form of an inactive prodrug, which, upon the cleavage of the linker is released in an active drug form. 
     
     
         10 . The composition of  claim 7 , wherein the release of the drug is facilitated by an enzymatic activity selected from: an enzymatic activity present on the surface the target cell, an enzymatic activity present inside the target cell, an enzymatic activity present in bodily fluids, an exogenous enzyme administered to a subject and an enzymatic activity facilitated by an enzyme encoded by a nucleic acid delivered to the target cell by the bacteriophage. 
     
     
         11 . The composition of  claim 7 , wherein the release of the drug is facilitated by at least one enzyme selected from the group consisting of: proteases, peptidases, esterases, amidases, glycosidases and lipases. 
     
     
         12 . The composition of  claim 7 , wherein the linker is selected from the group consisting of a branched linker and a dendrimer so that the composition can carry a plurality of drugs. 
     
     
         13 . The composition of  claim 7 , wherein the linker is an aminoglycoside. 
     
     
         14 . The composition of  claim 1  further comprising a pharmaceutically acceptable excipient or diluent. 
     
     
         15 . A bacteriophage comprising a coat protein having an amino acid sequence as set forth in SEQ ID NO:1 and analogs thereof. 
     
     
         16 . A bacteriophage-drug conjugate comprising an average of at least 1,000 drug molecules conjugated to the outer surface of the bacteriophage, with the bacteriophage displaying an exogenous targeting moiety that binds a cell surface molecule on a target cell. 
     
     
         17 . A method of treating a disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to  claim 1 . 
     
     
         18 . The method of  claim 17 , wherein the bacteriophage displays an exogenous targeting moiety which binds a cell surface molecule on a target cell. 
     
     
         19 . The method of  claim 17 , wherein the disease is selected from: a bacterial infection, a viral infection, a fungal infection, a yeast infection, a parasitic infection, a non-infectious disease or disorder, an autoimmune disease, a hyperproliferative disorder, restenosis, an angiogenesis-dependent disease and cancer. 
     
     
         20 . The method of  claim 17 , wherein the subject is a selected from the group consisting of mammals and non-mammalian animals. 
     
     
         21 . The method of  claim 20 , wherein the subject is human. 
     
     
         22 . The method of  claim 21 , wherein the bacteriophage comprises a nucleic acid molecule comprising an exogenous nucleic acid sequence that is transcribed and translated in the target cell. 
     
     
         23 . A method of treating a bacterial infestation, comprising exposing the bacteria to a bacteriophage-drug conjugate. 
     
     
         24 . A kit for generating a targeted bacteriophage-drug conjugate, comprising:
 (i) a bacteriophage-drug conjugate displaying a ligand capable of binding a targeting molecule that selectively binds a target molecule on a target cell; and   (ii) instructions for linking the targeting moiety to the ligand.