Therapeutic liposome composition and method of preparation

Abstract

Reagents for use in preparing a therapeutic liposome composition sensitized to a target cell are described. The reagents include a liposomal composition composed of pre-formed liposomes having an entrapped therapeutic agent and a plurality of targeting conjugates composed of a lipid, a hydrophilic polymer and a targeting ligand. The therapeutic, target-cell sensitized liposome composition is formed by incubating the liposomal composition with a selected conjugate.

1 . A plurality of targeting conjugates, each conjugate consisting essentially of (i) a lipid having a polar head group and a hydrophobic tail, (ii) a hydrophilic polymer having a proximal end and a distal end, said polymer attached at its proximal end to the head group of the lipid, and (iii) a targeting ligand having binding affinity for a receptor expressed on a cell attached to the distal end of the polymer, said plurality of conjugates in dried form.  
     
     
         2 . The conjugates of  claim 1 , wherein the lipid is selected from the group consisting of distearoyl phosphatidylethanolamine, distearoyl-phosphatidylcholine, monogalactosyl diacylglycerols and digalactosyl diacylglycerols.  
     
     
         3 . The conjugates of  claim 1 , wherein the hydrophilic polymer is selected from the group consisting of polyvinylpyrrolidone, polyvinylmethylether, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyloxazoline, polyhydroxypropylmethacrylamide, polymethacrylamide, polydimethylacrylamide, polyhydroxypropylmethacrylate, polyhydroxyethylacrylate, hydroxymethylcellulose, hydroxyethylcellulose, polyethyleneglycol, polyaspartamide and hydrophilic peptide sequences.  
     
     
         4 . The conjugates of  claim 1 , wherein the hydrophilic polymer is polyethylene glycol.  
     
     
         5 . The conjugates of  claim 4 , wherein the polyethylene glycol has a molecular weight between 500-5,000 daltons.  
     
     
         6 . The conjugates of  claim 1 , wherein the targeting ligand is an antibody or an antibody fragment.  
     
     
         7 . The conjugates of  claim 6 , wherein the antibody or antibody fragment is a humanized murine antibody.  
     
     
         8 . The conjugates of  claim 1 , wherein the targeting ligand binds to an extracellular domain of a growth factor receptor.  
     
     
         9 . The conjugates of  claim 8 , wherein the receptors are selected from the group consisting of c-erbB-2 protein product of the HER2/neu oncogene, epidermal growth factor receptor, basic fibroblast growth factor receptor and vascular endothelial growth factor receptor.  
     
     
         10 . The conjugates of  claim 1 , wherein the targeting ligand binds a receptor selected from the group consisting of E-selectin receptor, L-selectin receptor, P-selectin receptor, folate receptor, CD4 receptor, CD19 receptor, αβ integrin receptors and chemokine receptors.  
     
     
         11 . The conjugates of  claim 1 , wherein the targeting ligand binds a receptor on a malignant B-cell or T-cell, said receptor selected from the group consisting of CD19, CD20, CD22, CD4, CD7 and CD8.  
     
     
         12 . The conjugates of  claim 1 , wherein the targeting ligand is selected from the group consisting of folic acid, pyridoxal phosphate, vitamin B12, sialyl Lewis x , transferrin, epidermal growth factor, basic fibroblast growth factor, vascular endothelial growth factor, VCAM-1, ICAM-1, PECAM-1, RGD peptides and NGR peptides.  
     
     
         13 . The conjugates of  claim 1 , wherein the targeting ligand is selected from the group consisting of water soluble vitamins, apolipoproteins, insulin, galactose, Mac-1, PECAM-1/CD31, fibronectin, osteopontin, RGD sequences of matrix proteins, HIV GP 120/41 domain peptomers, GP120 C4 domain peptomers, T cell tropic isolates, SDF-1 chemokines, Macrophage tropic isolates, anti-cell surface receptor antibodies or fragments thereof, pyridoxyl ligands, biotin, RGD peptide mimetics, YIGSRG protein, a v B 5 , IL-8, anti-E-selectin Fab.  
     
     
         14 . The conjugates of  claim 13 , wherein the anti-cell surface receptor antibodies or fragments thereof is selected from the group consisting of anti-HER2/neu, anti-selectin and anti-VEGF pyridoxyl.  
     
     
         15 . The conjugates of  claim 13 , wherein the pyridoxyl ligand is selected from the group consisting of pyridoxal, pyridoxine, pyridoxamine, pyridoxal 5′-phosphate and N-(4′-pyridoxyl)amines.  
     
     
         16 . A targeting conjugate consisting essentially of (i) a lipid having a polar head group and a hydrophobic tail, (ii) a hydrophilic polymer having a proximal end and a distal end, said polymer attached at its proximal end to the head group of the lipid, and (iii) a targeting ligand attached to the distal end of the polymer, said targeting conjugate in dried form.  
     
     
         17 . The conjugate of  claim 16 , wherein the targeting ligand is an antibody or an antibody fragment.  
     
     
         18 . The conjugate of  claim 17 , wherein the antibody or antibody fragment is a humanized murine antibody.  
     
     
         19 . The conjugate of  claim 16 , wherein the targeting ligand binds to an extracellular domain of a growth factor receptor.  
     
     
         20 . The conjugate of  claim 19 , wherein the receptors are selected from the group consisting of c-erbB-2 protein product of the HER2/neu oncogene, epidermal growth factor receptor, basic fibroblast growth factor receptor and vascular endothelial growth factor receptor.  
     
     
         21 . The conjugate of  claim 16 , wherein the targeting ligand binds a receptor selected from the group consisting of E-selectin receptor, L-selectin receptor, P-selectin receptor, folate receptor, CD4 receptor, CD19 receptor, αβ integrin receptors and chemokine receptors.  
     
     
         22 . The conjugate of  claim 16 , wherein the targeting ligand binds a receptor on a malignant B-cell or T-cell, said receptor selected from the group consisting of CD19, CD20, CD22, CD4, CD7 and CD8.  
     
     
         23 . The conjugate of  claim 16 , wherein the targeting ligand is selected from the group consisting of folic acid, pyridoxal phosphate, vitamin B12, sialyl Lewis x , transferrin, epidermal growth factor, basic fibroblast growth factor, vascular endothelial growth factor, VCAM-1, ICAM-1, PECAM-1, RGD peptides and NGR peptides.  
     
     
         24 . The conjugate of claim  60 , wherein the targeting ligand is selected from the group consisting of water soluble vitamins, apolipoproteins, insulin, galactose, Mac-1, PECAM-1/CD31, fibronectin, osteopontin, RGD sequences of matrix proteins, HIV GP 120/41 domain peptomers, GP120 C4 domain peptomers, T cell tropic isolates, SDF-1 chemokines, Macrophage tropic isolates, anti-cell surface receptor antibodies or fragments thereof, pyridoxyl ligands, biotin, RGD peptide mimetics, YIGSRG protein, a v B 5 , IL-8, anti-E-selectin Fab.  
     
     
         25 . The conjugate of  claim 24 , wherein the anti-cell surface receptor antibodies or fragments thereof is selected from the group consisting of anti-HER2/neu, anti-selectin and anti-VEGF pyridoxyl.  
     
     
         26 . The conjugate of  claim 24 , wherein the pyridoxyl ligand is selected from the group consisting of pyridoxal, pyridoxine, pyridoxamine, pyridoxal 5′-phosphate and N-(4′-pyridoxyl)amines.