Targeted liposomes

Abstract

The present invention is in the field of drug delivery, and specifically, cationic liposome-based drug delivery. In embodiments, this invention provides methods of making ligand-targeted (e.g., antibody- or antibody fragment-targeted) liposomes useful for the delivery of liposomes to tumors, including brain tumors. In embodiments, the liposomes deliver temozolomide across the blood-brain barrier for treatment of primary or metastatic brain tumors. Additional cancers that can be treated with the liposomes include neuroendocrine tumors, melanoma, prostate, head and neck, ovarian, lung, liver, kidney, breast, urogenital, gastric, colorectal, cervical, vaginal, angiosarcoma, liposarcoma, rhabdomyosarcoma, choriocarcinoma, pancreatic, retinoblastoma and other types of cancer. In another embodiment the liposomes deliver melphalan for the treatment of multiple myeloma, other tumors of the blood or other solid tumors. In still other embodiments the liposomes can deliver other drugs such as pemetrexed or irinotecan for treatment of cancer or drugs including atropine for treatment of organophosphate poisoning.

1 . A method of preparing a targeted active agent cationic liposome complex, comprising:
 (a) preparing a lipid solution comprising one or more cationic lipids in ethanol;   (b) preparing a solution of an active agent selected from temozolomide, melphalan and atropine;   (c) mixing the lipid solution with the solution of active agent;   (d) injecting the mixture of lipid and active anent into an aqueous solution, thereby forming an active agent cationic liposome;   (e) mixing the active agent cationic liposome with a ligand to form the targeted active agent cationic liposome, wherein the ligand is directly complexed with, but not chemically conjugated to, the cationic liposome.   
     
     
         2 . The method of  claim 1 , wherein the ligand is an antibody, an antibody fragment or a protein. 
     
     
         3 . The method of  claim 2 , wherein the ligand is a single chain Fv antibody fragment. 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . The method of  claim 1 , wherein the lipid solution comprises 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). 
     
     
         7 . The method of  claim 1 , wherein the solution of active agent is prepared at a concentration of about 1 mM to about 200 mM. 
     
     
         8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein the molar ratio of lipid:active agent is about 0.1:1 to about 5:1. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . The method of  claim 1 , wherein the weight ratio of ligand:lipid is about 0.01:1 to about 0.5:10. 
     
     
         13 - 24 . (canceled) 
     
     
         25 . A method of treating cancer in a patient, comprising administering to the patient a targeted active agent cationic liposome complex, wherein the targeted active agent cationic liposome complex comprises:
 (a) a cationic liposome comprising 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);   (b) an active agent selected from temozolomide and melphalan; and   (c) a ligand directly complexed with, but not chemically conjugated to, the cationic liposome.   
     
     
         26 . The method of  claim 25 , wherein the ligand is an antibody, an antibody fragment or a protein. 
     
     
         27 . The method of  claim 26 , wherein the ligand is a single chain Fv antibody fragment. 
     
     
         28 . (canceled) 
     
     
         29 . The method of  claim 25 , wherein the active agent is administered to the patient at a dose of about 10 mg/m 2  to about 500 mg/m 2 . 
     
     
         30 . (canceled) 
     
     
         31 . The method of  claim 25 , wherein the molar ratio of lipid:active agent in the cationic liposomes is about 0.1:1 to about 5:1. 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . The method of  claim 25 , wherein the weight ratio of ligand:lipid in the cationic liposome is about 0.01:1 to about 0.5:10. 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . The method of  claim 25 , wherein the administration is intravenous (IV), intratumoral (IT), intralesional (IL), sublingual (SL), aerosal, percutaneous, oral, endoscopic, topical, intramuscular (IM), intradermal (ID), intraocular (IO), intraperitoneal (IP), transdermal (TD), intranasal (IN), intracereberal (IC), intraorgan (e.g. intrahepatic), slow release implant, or subcutaneous administration, or via administration using an osmotic or mechanical pump 
     
     
         38 . The method of  claim 25 , wherein the cancer is head and neck cancer, breast cancer, prostate cancer, pancreatic cancer, brain cancer, neuroendocrine cancer, cervical cancer, lung cancer, liver cancer, kidney cancer, liposarcoma, angiosarcoma, rhabdomyosarcoma, choriocarcinoma, melanoma, retinoblastoma, ovarian cancer, vaginal cancer, urogenital cancer, gastric cancer, colorectal cancer, multiple myeloma or a cancer of the blood. 
     
     
         39 . The method of  claim 38 , wherein the brain cancer is a glioma, astrocytoma or a glioblastoma. 
     
     
         40 . The method of  claim 25 , further comprising administering an additional different therapy to the patient in combination with the targeted active agent cationic liposome complex. 
     
     
         41 . The method of  claim 40 , wherein the additional different therapy comprises administration of a chemotherapeutic agent, a small molecule, radiation therapy or a nucleic acid-based therapy. 
     
     
         42 . The method of  claim 41 , wherein the nucleic acid-based therapy comprises administration of a cationic liposome complex comprising an antisense oligonucleotide, an siRNA, an miRNA, a plasmid DNA or an shRNA 
     
     
         43 - 54 . (canceled) 
     
     
         55 . A method of treating cancer in a patient, comprising administering to the patient a targeted active agent cationic liposome complex prepared by the method of  claim 1 . 
     
     
         56 . The method of  claim 55 , wherein the ligand is an antibody, an antibody fragment or a protein. 
     
     
         57 . The method of  claim 56 , wherein the ligand is a single chain Fv antibody fragment. 
     
     
         58 . (canceled) 
     
     
         59 . The method of  claim 55 , wherein active agent is administered to the patient at a dose of about 10 mg/m 2  to about 500 mg/m 2 . 
     
     
         60 . (canceled) 
     
     
         61 . The method of  claim 55 , wherein the molar ratio of lipid:active agent is about 0.1:1 to about 5:1. 
     
     
         62 . (canceled) 
     
     
         63 . (canceled) 
     
     
         64 . The method of  claim 55 , wherein the weight ratio of ligand:lipid is about 0.01:1 to about 0.5:10. 
     
     
         65 . (canceled) 
     
     
         66 . (canceled) 
     
     
         67 . The method of  claim 55 , wherein the administration is intravenous (IV), intratumoral (IT), intralesional (IL), aerosal, percutaneous, oral, endoscopic, topical, intramuscular (IM), intradermal (ID), sublingual (SL), intraocular (IO), intraperitoneal (IP), transdermal (TD), intranasal (IN), intracereberal (IC), intraorgan (e.g. intrahepatic), slow release implant, or subcutaneous administration, or via administration using an osmotic or mechanical pump 
     
     
         68 . The method of  claim 55 , wherein the cancer is head and neck cancer, breast cancer, prostate cancer, pancreatic cancer, brain cancer, neuroendocrine cancer, cervical cancer, lung cancer, liver cancer, kidney cancer, liposarcoma, angiosarcoma, rhabdomyosarcoma, choriocarcinoma, melanoma, retinoblastoma, ovarian cancer, vaginal cancer, urogenital cancer, gastric cancer, colorectal cancer, multiple myeloma or a cancer of the blood. 
     
     
         69 . (canceled) 
     
     
         70 . The method of  claim 55 , further comprising administering an additional different therapy to the patient in combination with the targeted active agent cationic liposome complex. 
     
     
         71 . The method of  claim 70 , wherein the additional different therapy comprises administration of a chemotherapeutic agent, a small molecule, radiation therapy or a nucleic acid-based therapy. 
     
     
         72 . The method of  claim 71 , wherein the nucleic acid-based therapy comprises administration of a cationic liposome complex comprising an antisense oligonucleotide, an siRNA, an miRNA, a plasmid DNA or an shRNA 
     
     
         73 - 153 . (canceled) 
     
     
         154 . A method of treating organophosphate poisoning in a patient, comprising administering to the patient a targeted atropine cationic liposome complex, wherein the targeted atropine cationic liposome complex comprises:
 (a) a cationic liposome comprising 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);   (b) atropine; and   (c) a ligand directly complexed with, but not chemically conjugated to, the cationic liposome.   
     
     
         155 . The method of  claim 154 , wherein the ligand is an antibody, an antibody fragment or a protein. 
     
     
         156 . The method of  claim 155 , wherein the ligand is a single chain Fv antibody fragment. 
     
     
         157 . (canceled) 
     
     
         158 . The method of  claim 154 , wherein the atropine is administered to the patient at a dose of about 1 mg to about 10 mg. 
     
     
         159 . (canceled) 
     
     
         160 . The method of  claim 154 , wherein the molar ratio of lipid:atropine in the cationic liposome: atropine is about 0.1:1 to about 5:1. 
     
     
         161 . (canceled) 
     
     
         162 . (canceled) 
     
     
         163 . The method of  claim 154 , wherein the weight ratio of ligand:lipid in the cationic liposome is about 0.01:1 to about 0.5:10. 
     
     
         164 . (canceled) 
     
     
         165 . (canceled) 
     
     
         166 . The method of  claim 154 , wherein the administration is intravenous (IV), intratumoral (IT), intralesional (IL), sublingual (SL), aerosal, percutaneous, oral, endoscopic, topical, intramuscular (IM), intradermal (ID), intraocular (IO), intraperitoneal (IP), transdermal (TD), intranasal (IN), intracereberal (IC), intraorgan (e.g. intrahepatic), slow release implant, or subcutaneous administration, or via administration using an osmotic or mechanical pump. 
     
     
         167 . The method of  claim 154 , wherein the liposome crosses the blood-brain barrier. 
     
     
         168 - 240 . (canceled) 
     
     
         241 . The method of  claim 25 , wherein the liposome crosses the blood-brain barrier.