Amphoteric Liposome Formulation

Abstract

The invention relates to compositions and methods to inhibit gene expression. In particular, the invention provides DNAi oligonucleotides sequestered by amphoteric liposomes for the treatment of cancer.

What is claimed is: 
     
         1 - 90 . (canceled) 
     
     
         91 . A mixture comprising:
 an amphoteric liposome and   an oligonucleotide comprising SEQ ID NO: 943 or 940,   wherein the amphoteric liposome comprises:
 a lipid mixture of
 at least one anionic and at least one cationic lipid, wherein at least one of the anionic and cationic lipids is pH-responsive, and 
 at least one neutral lipid, wherein the neutral lipids are independently selected from phosphatidylcholines, phosphatidylethanolamines or sterols; 
 
 wherein the lipid mixture contains a total of 5 to 95 mole % anionic and cationic lipids and 5 to 95 mole % neutral lipids. 
   
     
     
         92 . The mixture of  claim 91 , wherein the cationic lipids are selected from the group consisting of DMTAP, DPTAP, DOTAP, DC-Chol, TC-Chol, MoChol, HisChol, DPIM, CHIM, DORIE, DDAB, DAC-Chol, DOTMA, DOGS, (C18) 2 Gly + N,N-dioctadecylamido-glycine, CTAB, CPyC, DODAP, DOEPC and derivatives thereof, and the anionic lipids are selected from the group consisting of DOGSucc, POGSucc, DMGSucc, DPGSucc, DGSucc, DMPS, DPPS, DOPS, POPS, DMPG, DPPG, DOPG, POPG, DMPA, DPPA, DOPA, POPA, CHEMS, Cet-P and derivatives thereof, and wherein the phosphatidylcholines are selected from the group consisting of POPC, OPPC, natural or hydrogenated soy bean PC, natural or hydrogenated egg PC, DMPC, DPPC or DOPC and derivatives thereof, the phosphatidylethanolamines are selected from the group consisting of DOPE, DMPE, DPPE and derivatives thereof, and the sterol is cholesterol or a derivative thereof. 
     
     
         93 . The mixture of  claim 91 , wherein the cationic lipids are selected from the group consisting of DOTAP, DC-Chol, MoChol and HisChol, and the anionic lipids are selected from the group consisting of DMGSucc, DOPA, CHEMS and Cet-P. 
     
     
         94 . The mixture of  claim 91 , wherein the cationic lipid is MoChol and the anionic lipid is CHEMS. 
     
     
         95 . The mixture of  claim 94 , wherein the lipid mixture comprises 10 to 60 mole % of MoChol and 10 to 30 mole % of CHEMS. 
     
     
         96 . The mixture of  claim 91 , wherein the neutral lipids comprise POPC and DOPE. 
     
     
         97 . The mixture of  claim 96 , wherein the lipid mixture comprises 5 to 40 mole % of POPC and 20 to 50 mole % of DOPE. 
     
     
         98 . The mixture of  claim 91 , wherein the neutral lipids comprise POPC and Chol. 
     
     
         99 . The mixture of  claim 98 , wherein the lipid mixture comprises 10 to 50 mole % of POPC and 30 to 50 mole % of Chol. 
     
     
         100 . The mixture of  claim 91 , wherein the amphoteric liposome comprises POPC, DOPE, MoChol and CHEMS. 
     
     
         101 . The mixture of  claim 100 , wherein the lipid mixture comprises 3 to 20 mole % of POPC, 10 to 60 mole % of DOPE, 10 to 60 mole % of MoChol and 10 to 60 mole % of CHEMS. 
     
     
         102 . The mixture of  claim 100 , wherein the molar ratio of POPC/DOPE/MoChol/CHEMS is about 6/24/47/23. 
     
     
         103 . The mixture of  claim 100 , wherein the molar ratio of POPC/DOPE/MoChol/CHEMS is 15/45/20/20. 
     
     
         104 . The mixture of  claim 91 , wherein the amphoteric liposomes comprise a size between 50 and 500 nm. 
     
     
         105 . The mixture of  claim 91 , wherein the amphoteric liposome comprises a size between 80 and 300 nm. 
     
     
         106 . The mixture of  claim 91 , wherein the amphoteric liposome comprises a size between 90 and 200 nm. 
     
     
         107 . The mixture of any of  claim 91 , wherein the amphoteric liposome has an oligonucleotide concentration of at least 2 mg/ml at a lipid concentration of about 10 to 100 mM or less. 
     
     
         108 . The mixture of any of  claim 91 , wherein the oligonucleotide comprises SEQ ID NO:943. 
     
     
         109 . The mixture of any of  claim 91 , wherein the lipid mixture contains a total of 20 to 80 mole % anionic and cationic lipids and 20 to 80 mole % neutral lipids. 
     
     
         110 . The mixture of any of  claim 91 , wherein the lipid mixture contains a total of 30 to 70 mole % anionic and cationic lipids and 30 to 70 mole % neutral lipids. 
     
     
         111 . A method comprising:
 (a) providing the mixture of  claim 91  and an in vitro cell or animal capable of expressing the c-myc gene, and   (b) introducing the mixture into the in vitro cell or animal.   
     
     
         112 . The method of  claim 111 , wherein introducing the mixture results in a reduction of proliferation of the cell or induces cell death. 
     
     
         113 . The method of  claim 112 , wherein the cell is a cancer cell. 
     
     
         114 . The method of  claim 111 , wherein the mixture is introduced to the animal at a dosage of between 0.01 mg to 100 mg per kg of body weight. 
     
     
         115 . The method of  claim 111 , wherein the mixture is introduced to the animal one or more times per day. 
     
     
         116 . The method of  claim 111 , wherein the mixture is introduced to the animal continuously. 
     
     
         117 . The method of  claim 111 , wherein the mixture is introduced to the animal by one or more routes of administration selected from the group consisting of topical, pulmonary, intraocular, intranasal, parenteral, and a medical device. 
     
     
         118 . The method of  claim 111 , wherein the cell is in cell culture. 
     
     
         119 . The method of  claim 111 , wherein the animal has cancer which is selected from the group consisting of pancreatic cancer, colon cancer, breast cancer, bladder cancer, lung cancer, leukemia, prostate cancer, lymphoma, ovarian cancer and melanoma. 
     
     
         120 . A composition comprising the mixture of  claim 91 , which is to be introduced into a cell or an animal capable of expressing the c-myc gene. 
     
     
         121 . The composition of  claim 120 , wherein introducing the composition results in a reduction of proliferation of the cell or induces cell death. 
     
     
         122 . The composition of  claim 121 , wherein the cell is a cancer cell. 
     
     
         123 . The composition of  claim 120 , which is introduced into the animal at a dosage of between 0.01 mg to 100 mg per kg of body weight. 
     
     
         124 . The composition of  claim 120 , which is introduced into the animal one or more times per day. 
     
     
         125 . The composition of  claim 120 , which is introduced into the animal continuously. 
     
     
         126 . The composition of  claim 120 , wherein the animal has a cancer which is selected from the group consisting of pancreatic cancer, colon cancer, breast cancer, bladder cancer, lung cancer, leukemia, prostate cancer, lymphoma, ovarian cancer and melanoma. 
     
     
         127 . A mixture comprising amphoteric liposome and an oligonucleotide comprising SEQ ID NO:943 or 940 wherein the liposome comprises POPC, DOPE, MoChol and CHEMS in the molar ratio of POPC/DOPE/MoChol/CHEMS of 6/24/47/23.