Modified nucleoside, nucleotide, and nucleic acid compositions

Abstract

The present disclosure provides, inter alia, formulation compositions comprising modified nucleic acid molecules which may encode a protein, a protein precursor, or a partially or fully processed form of the protein or a protein precursor. The formulation composition may further include a modified nucleic acid molecule and a delivery agent. The present invention further provides nucleic acids useful for encoding polypeptides capable of modulating a cell's function and/or activity.

1 - 54 . (canceled) 
     
     
         55 . A pharmaceutical composition comprising:
 a plurality of lipoplexes comprising two or more cationic lipids and an mRNA encoding a polypeptide, wherein the mRNA comprises:   (i) at least one 5′-cap structure;   (ii) a 5′-UTR;   (iii) an open reading frame encoding the polypeptide and consisting of nucleotides including a modified uridine, cytosine, adenine, and guanine;   (iv) a 3′-UTR; and   (v) a poly-A region of least 100 nucleotides in length.   
     
     
         56 . The pharmaceutical composition of  claim 55 , wherein the modified uridine is pseudouridine or 1-methyl-pseudouridine. 
     
     
         57 . The pharmaceutical composition of  claim 55 , wherein the modified uridine is 1-methyl-pseudouridine. 
     
     
         58 . The pharmaceutical composition of  claim 55 , 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. 
     
     
         59 . The pharmaceutical composition of  claim 58 , wherein the at least one 5′-cap structure is cap0, cap1, or ARCA. 
     
     
         60 . The pharmaceutical composition of  claim 55 , wherein the poly-A tail is at least 160 nucleotides in length. 
     
     
         61 . The pharmaceutical composition of  claim 55 , wherein the 5′-UTR comprises a Kozak sequence. 
     
     
         62 . A method of expressing a secreted protein in a mammalian subject, the method comprising administering a pharmaceutical composition of  claim 55  to the subject. 
     
     
         63 . The method of  claim 62 , wherein the method comprises administering about 0.05 to about 0.5 mg/kg of polynucleotide. 
     
     
         64 . The method of  claim 62 , wherein the administration is intramuscular administration. 
     
     
         65 . The method of  claim 62 , wherein the administration is intravenous administration. 
     
     
         66 . The method of  claim 62 , wherein the administration is subcutaneous administration.