Delivery of nucleic acid-like compounds

Abstract

Provided herein is a process for preparing a microparticulate complex of a particle-forming component (“PFC”) and a nucleic acid-like component (“NAC”) in a monophasic composition comprising water and a water-miscible organic solvent. Also provided is a microparticulate complex that comprises a particle-forming component complexed to a nucleic acid-like component, and a composition comprising water and the microparticulate complex. Further provided is a method of delivery of a nucleicacid-like component to a cell or to a patient by contacting the cell with or administering to the patient a composition comprising water and the microparticulate complex described herein. Still further, a charge-changing composition represented by the formula A-X—B is provided.

1 .- 22 . (canceled) 
     
     
         23 . A microparticulate complex that comprises a particle-forming component (“PTC”) complexed with a nucleic acid-like component forming an approximately spherical particle, wherein the particle-forming component encloses an interior of the particle containing the nucleic acid-like component and free of chloroform, and less than about 50% of the lipid-enclosed interior volume of the microparticulate complex contains free water. 
     
     
         24 . The complex of  claim 23 , wherein the particle-forming component comprises a cationic lipid. 
     
     
         25 . The complex of  claim 23 , wherein the particle ranges in size between about 40 nm to about 1500 nm. 
     
     
         26 . The complex of  claim 25 , wherein the particle ranges in size from about 60 nm to about 350 nm. 
     
     
         27 . The complex of  claim 23 , wherein the lipid-enclosed interior volume has less than 20% of the volume containing free water. 
     
     
         28 . (canceled) 
     
     
         29 . The microparticulate complex of  claim 23 , wherein a ligand is conjugated to the complex. 
     
     
         30 . The microparticulate complex of  claim 29 , wherein the ligand is conjugated to a lipophilic anchor through a hydrophilic polymer linker. 
     
     
         31 . The complex of  claim 23 , wherein an amphiphilic lipid is combined with the PFC. 
     
     
         32 . The complex of  claim 31 , wherein the amphiphilic lipid is a hydrophilic polymer-lipid conjugate. 
     
     
         33 . The complex of  claim 32 , wherein the hydrophilic polymer is a poly(ethylene glycol) having a molecular weight of about 250 to about 20,000 and the lipid is a phospholipid, a sphingolipid, a diacylglyerol, or a sterol. 
     
     
         34 . The complex of  claim 33 , wherein the amphiphilic lipid is present in an amount less than about 5.0 mol. %. 
     
     
         35 . The complex of  claim 31 , wherein the amphiphilic lipid is a polyoxyethylene ether of a fatty alchol, a polyoxyethylene ester of a fatty acid, or a polyoxyethylene ether of a polyol derivatized with a fatty alcohol or a fatty acid. 
     
     
         36 . A composition comprising water and particles of a microparticulate complex of  claim 23 . 
     
     
         37 .- 94 . (canceled) 
     
     
         95 . A microparticulate complex comprising a nucleic acid component and a membrane encapsulating the nucleic acid component,
 wherein said membrane comprises at least one cationic lipid, at least one neutral lipid, and   optionally at least one amphiphilic lipid,   the enclosed interior of the microparticulate complex contains the nucleic acid component and an aqueous content,   wherein the aqueous content in said microparticulate complex (excluding the nucleic acid component) has an ionic strength of below that of 50 mM NaCl.   
     
     
         96 . The complex of  claim 95 , wherein the particle ranges in size between about 40 nm to about 1500 nm. 
     
     
         97 . The complex of  claim 96 , wherein the particle ranges in size from about 60 nm to about 350 nm. 
     
     
         98 . The complex of  claim 95 , wherein the lipid-enclosed interior volume has less than 20% of the volume containing free water. 
     
     
         99 . The microparticulate complex of  claim 95 , wherein a ligand is conjugated to the complex. 
     
     
         100 . The microparticulate complex of  claim 99 , wherein the ligand is conjugated to a lipophilic anchor through a hydrophilic polymer linker. 
     
     
         101 . The complex of  claim 99 , comprising an amphiphilic lipid. 
     
     
         102 . A composition comprising particles of a microparticulate complex of  claim 95  in an aqueous medium. 
     
     
         103 . A microparticulate complex of lipid and nucleic acid, said complex prepared by a process comprising: combining a lipid composition comprising cationic and non-cationic lipids, and a nucleic acid component, in an aqueous/organic solvent system comprising water and less than 55% by volume of a water-miscible organic solvent, wherein the aqueous component of the aqueous/organic solvent system has an ionic strength of less than that of about 50 mM NaCl, to form a mixture which is heated to a temperature of from 30° to 80° C., forming a monophasic system wherein the aqueous/organic solvent system is a single liquid phase and wherein the lipid composition and the nucleic acid component are independently molecularly or micellarly soluble in he single liquid phase, and reducing the amount of the organic solvent in the monophasic system so that the microparticulate complex is formed. 
     
     
         104 . The microparticulate complex of  claim 103 , wherein, in the preparation of the microparticulate complex, prior to combining the lipid composition and the nucleic acid component, the cationic lipid is dissolved in a mixture of water and organic solvent and the nucleic acid component is in an aqueous composition. 
     
     
         105 . The microparticulate complex of  claim 103 , wherein the microparticulate complex is in the form of particles that are approximately spherical and range in size from about 60 nm to about 350 nm. 
     
     
         106 . The microparticulate complex of  claim 103 , wherein, subsequent to the formation of the microparticulate complex, the microparticulate complex is separated from the liquid phase. 
     
     
         107 . The microparticulate complex of  claim 106 , wherein the separation is carried out by size exclusion chromatography. 
     
     
         108 . The microparticulate complex of  claim 103 , in which a ligand is attached to the surfaces of the particles. 
     
     
         109 . The microparticulate complex of  claim 108 , wherein the ligand is conjugated to a lipophilic anchor through a hydrophilic polymer linker. 
     
     
         110 . The microparticulate complex of  claim 109 , wherein the hydrophilic polymer is a poly(ethylene glycol) having a molecular weight of about 250 to about 20,000 and the lipid is a phospholipid, a sphingolipid, a diacylglyerol, or a sterol. 
     
     
         111 . The process of  claim 103 , wherein particles comprise an amphiphilic lipid that is present in an amount less than about 5.0 mol. %. 
     
     
         112 . The process of  claim 103 , wherein the water-miscible organic solvent is a water-miscible alcohol or ether. 
     
     
         113 . The microparticulate complex of  claim 103  wherein aqueous/organic solvent system consists essentially of water and less than 55% by volume of a water-miscible organic solvent. 
     
     
         114 . The microparticulate complex of  claim 112 , wherein, in the preparation of the microparticulate complex, an amphiphilic lipid is mixed with the mixture when the lipid composition and the nucleic acid component are combined or when the amount of the alcohol or ether in the monophasic system is reduced. 
     
     
         115 . The microparticulate complex of  claim 114 , wherein the amphiphilic lipid is a hydrophilic polymer-lipid conjugate, a polyoxethylene ether of a fatty alcohol, a polyoxyethylene ester of a fatty acid, or a polyethylene ether of a polyol derivatized with a fatty alcohol or a fatty acid.