Apparatus and methods for making nanosomes loaded with nucleic acid

Abstract

Embodiments of the present invention are directed to an apparatus and methods for making nucleic acid loaded nanosomes. One embodiment of the present invention directed to an apparatus comprises a first containment means for containing a mixture of an aqueous solution of nucleic acid and a phospholipid solution with a supercritical, critical or near critical fluid. The apparatus further comprises injection means in fluid communication with said first containment means for receiving the mixture and releasing the mixture as a stream into a decompression liquid. The apparatus further comprises a decompression vessel in fluid communication with the injection means for holding a decompression liquid and receiving the mixture as a stream. The stream forms one or more nanosomes loaded with a nucleic acid in the decompression liquid.

1 . An apparatus for making a nanosome loaded with a siRNA comprising:
 a. a first containment means for containing a mixture of an aqueous solution of a siRNA and a phospho lipid solution in a supercritical, critical or near critical fluid;   b. injection means in fluid communication with said first containment means for receiving said mixture and releasing said mixture as a stream into a decompression liquid; and   c. a decompression vessel in fluid communication with said injection means for holding a decompression liquid and receiving said mixture as a stream in said decompression liquid, said stream forming one or more nanosomes loaded with the siRNA.   
     
     
         2 . The apparatus of  claim 1  further comprising a circulation loop in fluid communication with said first containment means, said circulation loop for forming said phospholipid solution in the supercritical, critical or near critical fluid. 
     
     
         3 . The apparatus of  claim 2  wherein said circulation loop has a solids vessel for holding a phospholipid and forming the phospholipid solution in the supercritical, critical or near critical fluid. 
     
     
         4 . The apparatus of  claim 3  wherein said circulation loop has a mixing chamber in communication with said solids vessel for receiving said phospholipid solution in the supercritical, critical or near critical fluid. 
     
     
         5 . The apparatus of  claim 2  wherein said circulation loop is in fluid communication with a source of the supercritical, critical or near critical fluid. 
     
     
         6 . The apparatus of  claim 3  wherein said circulation loop has return means in fluid communication with said mixing chamber and said solids vessel for returning the phospholipid solution in the supercritical, critical or near critical fluid from said mixing chamber to said solids vessel increase the phospholipid content of said phospholipid solution in the supercritical, critical or near critical fluid. 
     
     
         7 . The apparatus of  claim 6  wherein said circulation loop has one or more pumps to move said phospholipid solution in the supercritical, critical or near critical fluid through said solids vessel and mixing chamber. 
     
     
         8 . The apparatus of  claim 1  wherein said first containment means is in fluid communication with a source of the siRNA. 
     
     
         9 . The apparatus of  claim 8  wherein said source of the siRNA holds the siRNA in a buffer. 
     
     
         10 . The apparatus of  claim 9  wherein said buffer is a low ionic strength buffer. 
     
     
         11 . The apparatus of  claim 1  wherein said containment means is an inline mixer. 
     
     
         12 . As an article of manufacture, a nanosome comprising a phospholipid and a siRNA with trace amounts of a low ionic strength buffer.