Dry powder formulations for messenger rna

Abstract

The present invention provides stable, dry powder messenger RNA formulations for therapeutic use, and methods of making and using the same.

1 . A dry powder formulation for delivery of cystic fibrosis conductance regulator (CFTR) messenger RNA (mRNA) comprising a plurality of spray-dried particles comprising
 mRNA encoding a CFTR protein;   one or more lipids, and   one or more polymers.   
     
     
         2 . (canceled) 
     
     
         3 . The dry powder formulation of  claim 1 , wherein the one or more lipids and the one or more polymers are present in one or more lipid nanoparticles (LNPs) encapsulating the mRNA encoding the CFTR protein. 
     
     
         4 . The dry powder formulation of  claim 1 , wherein the CFTR mRNA has an integrity of 90% or greater. 
     
     
         5 .- 7 . (canceled) 
     
     
         8 . The dry powder formulation of  claim 1 , wherein the dry powder formulation is inhalable or nebulizable upon reconstitution. 
     
     
         9 . (canceled) 
     
     
         10 . The dry powder formulation of  claim 1 , wherein the one or more polymers constitute at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the combined weight of the lipids and polymers. 
     
     
         11 .- 12 . (canceled) 
     
     
         13 . The dry powder formulation of  claim 1 , wherein the one or more polymers are selected from a group consisting of chitosan, poly(lactic acid) (PLA), poly(lactic-co-glycolic acid) (PLGA), poly(q-caprolactone (PCL), poly amido amines, polyesters, polycarbonates, poly(hydroxyalkyl L-asparagine), poly(hydroxyalkyl L-glutamine), poly(2-alkyloxazoline) acrylates, modified acrylates and polymethacrylate based polymers, poly-N-(2-hydroxyl-propyl)methacrylamide, poly-2-(methacryloyloxy)ethyl phosphorylcholines, poly(2-(methacryloyloxy)ethyl phosphorylcholine), and poly(dimethylaminoethyl methylacrylate) (pDMAEMA). 
     
     
         14 . The dry powder formulation of  claim 1 , wherein the one or more polymers comprise a polymethacrylate based polymer or Eudragit EPO. 
     
     
         15 . (canceled) 
     
     
         16 . The dry powder formulation of  claim 3 , wherein the one or more LNPs encapsulating mRNA have a lipid:mRNA (N/P) ratio ranging from 1 to 20, 1-15, 1-10, 2-8, 2-6, or 2-4. 
     
     
         17 . (canceled) 
     
     
         18 . The dry powder formulation of  claim 3 , wherein the LNPs have an encapsulation efficiency of 80% or greater. 
     
     
         19 . The dry powder formulation of  claim 1 , wherein the one or more lipids comprise a cationic lipid, and wherein the cationic lipid is selected from the group consisting of C12-200, DOTAP (1,2-dioleyl-3-trimethytammonium propane), DODAP (1,2-dioleyl-3-dimethylammonium propane), DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane), DLinDMA, DLin-KC2-DMA, HGT4003, cKK-E12, ICE, and combinations thereof. 
     
     
         20 .- 22 . (canceled) 
     
     
         23 . The dry powder formulation of  claim 19 , wherein the cationic lipid constitutes about 25-50% of the total lipids in LNPs by molar. 
     
     
         24 . The dry powder formulation of  claim 19 , wherein the one or more lipids comprise a PEG-modified lipid. 
     
     
         25 .- 33 . (canceled) 
     
     
         34 . The dry powder formulation of  claim 1 , further comprising a pharmaceutically acceptable excipient selected from the group consisting of esters, urethanes, phosphoesters, phosphazenes, amino acids, collagen, chitosan, polysaccharides, albumin, surfactants, buffers, salts, and combinations thereof. 
     
     
         35 . The dry powder formulation of  claim 1 , wherein the surfactant is selected from the group consisting of CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate), phospholipids, phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine, sphingomyelins, Octaethylene glycol monododecyl ether, Pentaethylene glycol monododecyl ether, Triton X-100, Cocamide monoethanolamine, Cocamide diethanolamine, Glycerol monostearate, Glycerol monolaurate, Sorbitan moonolaureate, Sorbitan monostearate, Tween 20, Tween 40, Tween 60, Tween 80, Alkyl polyglucosides, and a poloxamer. 
     
     
         36 . (canceled) 
     
     
         37 . The dry powder formulation of  claim 1 , wherein the CFTR mRNA constitutes about 1-20%, 1-15%, 1-10%, 1-8%, 1-6%, 1-5%, 5-15%, or 5-10% of the total weight of the spray-dried particles. 
     
     
         38 . (canceled) 
     
     
         39 . A method of delivering cystic fibrosis conductance regulator (CFTR) messenger RNA (mRNA) for in vivo expression comprising a step of administering to a subject in need thereof the dry powder formulation of  claim 1 . 
     
     
         40 .- 44 . (canceled) 
     
     
         45 . A dry powder formulation for delivery of messenger RNA (mRNA) comprising a plurality of spray-dried particles comprising
 mRNA encoding a protein or a peptide;   one or more lipids, and   one or more polymers.   
     
     
         46 . A dry powder formulation for delivery of messenger RNA (mRNA) of  claim 18 , comprising a plurality of spray-dried particles comprising
 a. one or more lipid nanoparticles (LNPs) encapsulating mRNA encoding a peptide or polypeptide, and   b. one or more polymers.   
     
     
         47 . A dry powder formulation for delivery of messenger RNA (mRNA) of  claim 18 , comprising a plurality of spray-dried particles comprising one or more nanoparticles encapsulating mRNA encoding a peptide or polypeptide, the nanoparticles comprising
 a. one or more lipids, and   b. one or more polymers.   
     
     
         48 .- 84 . (canceled) 
     
     
         85 . A method for manufacturing a dry powder formulation, the method comprising:
 providing a mixture comprising an mRNA, one or more lipids and a polymer; and   spray-drying the mixture to form a plurality of particles.   
     
     
         86 .- 96 . (canceled)