Method for producing a protein of interest in a primate
View Patent ↗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. A method for producing a protein of interest in a primate comprising delivering to cells of said primate a modified mRNA comprising SEQ ID NO: 9, wherein the modified mRNA is formulated in a lipid nanoparticle.
2. The method of claim 1 , wherein the modified mRNA encodes erythropoietin.
3. The method of claim 1 , wherein the modified mRNA has a 5′ cap structure, wherein the 5′ cap structure is Cap1.
4. The method of claim 3 , wherein the modified mRNA has a polyA tail, wherein the polyA tail is approximately 160 nucleotides in length.
5. The method of claim 1 wherein the lipid nanoparticle has a final total molar ratio of 50 mol % cationic lipid, 10 mol % DSPC, 37-38.5 mol % Cholesterol and 1.5-3 mol % of PEG lipid.
6. The method of claim 5 , wherein the lipid nanoparticle has a 20:1 weight ratio of total lipid to the modified mRNA.
7. The method of claim 6 , wherein the cationic lipid is DLin-KC2-DMA.
8. The method of claim 7 , wherein the PEG lipid is PEG-c-DOMG.
9. The method of claim 8 , wherein the final total molar ratio of PEG-c-DOMG is 1.5% and wherein cholesterol has a final total molar ratio of 38.5 mol %.
10. The method of claim 9 , wherein the protein of interest is detectable at 24 hours after contacting said primate.
11. The method of claim 8 , wherein the primate is contacted with modified mRNA at a dose selected from the group consisting of 0.005 mg/kg, 0.05 mg/kg and 0.5 mg/kg.