Modified polynucleotides for the production of biologics and proteins associated with human disease

Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

1 - 92 . (canceled) 
     
     
         93 . A pharmaceutical composition comprising:
 a plurality of lipid nanoparticles encapsulating a polynucleotide, wherein the lipid nanoparticles comprise a cationic lipid, a neutral lipid, cholesterol, and a PEGylated lipid, wherein the plurality of lipid nanoparticles has a mean particle size of between 80 nm and 160 nm, and   wherein the polynucleotide comprises:   (i) at least one 5′-cap structure;   (ii) a 5′-UTR;   (iii) an open reading frame encoding a secreted protein and consisting of nucleotides selected from uridine, cytidine, adenosine, and guanosine;   (iv) a 3′-UTR; and   (v) a poly-A region of least 100 nucleotides in length.   
     
     
         94 . The pharmaceutical composition of  claim 93 , wherein the cationic lipid is a biodegradable cationic lipid. 
     
     
         95 . The pharmaceutical composition of  claim 94 , wherein the biodegradable cationic lipid comprises an ester linkage. 
     
     
         96 . The pharmaceutical composition of  claim 95 , wherein the biodegradable cationic lipid comprises DLin-DMA with an internal ester, DLin-DMA with a terminal ester, DLin-MC3-DMA with an internal ester, or DLin-MC3-DMA with a terminal ester. 
     
     
         97 . The pharmaceutical composition of  claim 93 , wherein the at least one 5′-cap structure is cap0, cap1, ARCA, inosine, N1-methyl-guanosine, 2′-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, or 2-azido-guanosine. 
     
     
         98 . The pharmaceutical composition of  claim 97 , wherein the at least one 5′-cap structure is cap0, cap1, or ARCA. 
     
     
         99 . The pharmaceutical composition of  claim 93 , wherein the 3′-UTR is an alpha-globin 3′-UTR. 
     
     
         100 . The pharmaceutical composition of  claim 93 , wherein the poly-A region is at least 160 nucleotides in length. 
     
     
         101 . The pharmaceutical composition of  claim 93 , wherein the 5′-UTR comprises a Kozak sequence. 
     
     
         102 . The pharmaceutical composition of  claim 93 , wherein the plurality of lipid nanoparticles has a mean PDI of between 0.02 and 0.2. 
     
     
         103 . The pharmaceutical composition of  claim 93 , wherein the plurality of lipid nanoparticles has a mean lipid to polynucleotide ratio (wt/wt) of between 10 and 20.