Systemic delivery of serum stable plasmid lipid particles for cancer therapy

Abstract

The present invention relates to methods and compositions for treating a neoplasia in a mammal.

1 . A method of treating a tumor in a mammal, said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion, 
 wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal;    wherein said lipid portion of said nucleic-acid lipid particle comprises a cationic lipid, a neutral lipid, and a lipid conjugate that prevents aggregation during formulation:    wherein cells of said tumor are responsive to said nucleic acid; and    wherein cells of said tumor are transfectable by said nucleic acid.    
     
     
         2 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein said nucleic acid comprises an expressible gene.  
     
     
         3 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein said expressible gene encodes a member selected from the group consisting of therapeutic polypeptides and therapeutic polynucleotides.  
     
     
         4 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein said gene is exogenous.  
     
     
         5 . A method of treating a tumor in a mammal in accordance with  claim 3 , wherein said gene is a member selected from the group consisting of genes encoding suicide enzymes, toxins and ribozymes.  
     
     
         6 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein said gene encodes a member selected from the group consisting of herpes simplex virus thymidine kinase (HSV-TK), cytosine deaminase, xanthine-guaninephosphoribosyl transferase, purine nucleoside phosphorylase, cytochrome P450 2B1 and analogs thereof.  
     
     
         7 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein said gene is homologous to a gene in the mammal.  
     
     
         8 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein said gene encodes a member selected from the group consisting of proto-oncogenes, cytokines, immune stimulatory proteins and anti-angiogenic proteins.  
     
     
         9 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein said gene is a member selected from the group consisting of IL-2, IL-12, IL-15 and GM-CSF.  
     
     
         10 . A method of treating a tumor in a mammal in accordance with  claim 2 , wherein a therapeutically effective amount of said gene is generated at said tumor.  
     
     
         11 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein said nucleic acid-lipid particle comprises a protonatable lipid having a pKa in the range of about 4 to about 11.  
     
     
         12 . A method of treating a tumor in a mammal in accordance with  claim 11 , wherein said protonatable lipid is a member selected from the group consisting of DODAC, DODAP, DODMA, DOTAP, DOTMA, DC-Chol, DMRIE, DSDAC and mixtures thereof.  
     
     
         13 . (canceled)  
     
     
         14 . A method of treating a tumor in a mammal in accordance with  claim 13 , wherein said lipid conjugate is a member selected from the group consisting of PEG-lipids and PAO-lipids.  
     
     
         15 . A method of treating a tumor in a mammal in accordance with  claim 13 , wherein said lipid conjugate is reversibly associated with an outer lipid monolayer, and wherein said lipid conjugate exchanges out of said outer lipid monolayer at a rate faster than PEG-CerC20.  
     
     
         16 . A method of treating a neoplasia tumor in a mammal in accordance with  claim 1 , wherein said nucleic acid-lipid particle is substantially devoid of detergents and organic solvents.  
     
     
         17 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein a therapeutically effective amount of said nucleic acid-lipid particle accumulates at said neoplasia tumor.  
     
     
         18 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein a therapeutic effect is detected at the site of said tumor.  
     
     
         19 . A method of treating a tumor in a mammal in accordance with  claim 17 , wherein said therapeutically effective amount comprises greater than about 0.5% of an administered dose.  
     
     
         20 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein said nucleic acid-lipid particle has a diameter of about 50 nm to about 200 nm.  
     
     
         21 . A method of treating a tumor in a mammal in accordance with  claim 20 , wherein said nucleic acid-lipid particle has a diameter of about 60 ni to about 130 nm.  
     
     
         22 . A method of treating a tumor in a mammal in accordance with  claim 20 , wherein said nucleic acid-lipid particles are of a uniform size.  
     
     
         23 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein said nucleic acid-lipid particle has a nucleic acid to lipid ratio of greater than about 3 mg nucleic acid to mmole of lipid.  
     
     
         24 . A method of treating a tumor in a mammal in accordance with  claim 23 , wherein said particle has a nucleic acid to lipid ratio of greater than about 14 mg nucleic acid to mmole of lipid.  
     
     
         25 . A method of treating a tumor in a mammal in accordance with  claim 23 , wherein said particle has a nucleic acid to lipid ratio of greater than about 25 mg nucleic acid to mmole of lipid.  
     
     
         26 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein said nucleic acid remains at least 90% intact when said particle containing about 1 μg DNA is treated with about 100 U DNAse 1 in digestion buffer at 37° C. for 30 min.  
     
     
         27 . A method of treating a tumor in a mammal in accordance with  claim 1 , further comprising administering a chemotherapeutic agent.  
     
     
         28 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein said administering is performed at least once per eight weeks.  
     
     
         29 . A method of sensitizing a neoplastic cell to a compound, said method comprising: 
 a) transfecting said neoplastic cell with a serum-stable nucleic acid-lipid particle encoding a gene-product comprising a nucleic acid that is fully encapsulated within a lipid, wherein administration of said nucleic acid-lipid particle is by injection at an injection site that is distal to said neoplastic cell; and    b) delivering to said cell a first compound which is processed by said gene-product into a second compound, wherein said cell is more sensitive to said second compound than said first compound.    
     
     
         30 . A method of sensitizing a neoplastic cell in accordance with  claim 29  wherein said first compound is formulated in a lipid.  
     
     
         31 . A method of sensitizing a neoplastic cell in accordance with  claim 29  wherein said gene product is a member selected from the group consisting of therapeutic polypeptides and therapeutic polynucleotides.  
     
     
         32 . A method of sensitizing a neoplastic cell in accordance with  claim 29  wherein said gene product is a member selected from the group consisting of suicide enzymes, toxins and ribozymes.  
     
     
         33 . A method of sensitizing a neoplastic cell in accordance with  claim 29  wherein said gene product is a member selected from the group consisting of herpes simplex virus thymidine kinase (HSV-TK), cytosine deaminase, xanthine-guaninephosphoribosyl transferase, purine nucleoside phosphorylase, cytochrome P450 2B1 and analogs thereof.  
     
     
         34 . A method of sensitizing a neoplastic cell in accordance with  claim 29  wherein a therapeutic effect is detected at the site of said neoplasia cell.  
     
     
         35 . A method of treating a tumor in a mammal, in accordance with  claim 5 , wherein said gene encodes a suicide enzyme and said method further comprises administering a prodrug.  
     
     
         36 . A method of treating a tumor in a mammal in accordance with  claim 35  , wherein said prodrug is administered after the serum-stable nucleic acid-lipid particle.  
     
     
         37 . A method of treating a tumor in a mammal in accordance with  claim 35 , wherein said prodrug is administered before the serum-stable nucleic acid-lipid particle.  
     
     
         38 . A method of treating a tumor in a mammal in accordance with  claim 9 , further comprising administering a chemotherapeutic agent.  
     
     
         39 . A method of treating a tumor in a mammal in accordance with  claim 38 , wherein the chemotherapeutic agent is administered after the serum-stable nucleic acid-lipid particle.  
     
     
         40 . A method of treating a tumor in a mammal in accordance with  claim 38 , wherein the chemotherapeutic agent is administered before the serum-stable nucleic acid-lipid particle.  
     
     
         41 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein the cationic lipid is DODAC.  
     
     
         42 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein the neutral lipid is DOPE.  
     
     
         43 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein the lipid portion further comprises a PEG-lipid.  
     
     
         44 . A method of treating a tumor in a mammal in accordance with  claim 1 , wherein the lipid portion further comprises cholesterol.  
     
     
         45 . A method of treating a tumor in a mammal, said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion, 
 wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal; and    wherein said tumor is responsive to the gene product- of the nucleic acid; and    wherein said nucleic-acid lipid particle comprises a cationic lipid, a neutral lipid, and a lipid conjugate that prevents aggregation during formulation.    
     
     
         46 . A method of treating a tumor in a mammal, said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion, 
 wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal;    wherein cells of said tumor are transfectable by said nucleic acid-lipid particle; and    and wherein said lipid portion of said nucleic-acid lipid particle comprises a cationic lipid, a neutral lipid, and a lipid conjugate that prevents aggregation during formulation.    
     
     
         47 . The method of  claim 45 , wherein said nucleic acid encodes a member selected from the group consisting of: suicide enzymes, toxins, tumor suppressor genes, and cytokines.  
     
     
         48 . The method of  claim 45 , wherein said nucleic acid encodes a suicide enzyme; and said method further comprises administering a prodrug.  
     
     
         49 . The method of  claim 45 , wherein said nucleic acid encodes a toxin.  
     
     
         50 . The method of  claim 45 , wherein said nucleic acid encodes a tumor suppressor protein.  
     
     
         51 . The method of  claim 45 , wherein said nucleic acid encodes a cytbkine.  
     
     
         52 . The method of  claim 48 , wherein the suicide enzyme is a member selected from the group consisting of: HSV-TK, purine nucleoside phosphorylase, and cytosine deaminase.  
     
     
         53 . The method of  claim 48 , wherein the tumor is a melanoma.  
     
     
         54 . The method of  claim 48 , wherein the tumor is a colorectal tumor.  
     
     
         55 . The method of  claim 48 , wherein the tumor is a sarcoma.  
     
     
         56 . The method of  claim 49 , wherein the toxin is Pseudomonas exotoxin.  
     
     
         57 . The method of  claim 50 , wherein the tumor suppressor protein is apoptin.  
     
     
         58 . The method of  claim 51 , wherein the cytokine is IL-12.  
     
     
         59 . The method of  claim 39 , wherein delivery of the serum-stable nucleic acid-lipid particle is intravenous.