Pulmonary delivery of mRNA to non-lung target cells
Compositions comprising mRNA formulated for pulmonary administration and related methods for delivery of the mRNA and/or encoded protein to a non-lung cell or tissue. The compositions and methods may be used to prevent or ameliorate the symptoms of diseases associated with the mRNA encoded protein.
1 . A composition comprising one or more mRNA encoding an antibody and a lipid carrier vehicle, wherein the composition is formulated for pulmonary delivery, wherein the antibody is a full-length antibody, Fab, Fv, or single-chain Fv, wherein the lipid carrier vehicle comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids, wherein the cationic lipid comprises a molar ratio of about 20% to about 70% of the total lipid present in the lipid carrier vehicle, wherein the composition is contained in a device suitable for delivery to the lungs.
2 . The composition of claim 1 , wherein the one or more cationic lipids are selected from the group consisting of C12-200, HGT4003, HGT5000, HGT5001, ICE, DLinKC2-DMA, DODAP, DODMA, DLinDMA, CLinDMA, and combinations thereof.
3 . The composition of claim 1 , wherein the one or more non-cationic lipids are selected from the group consisting of distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), cholesterol, or a mixture thereof.
4 . The composition of claim 1 , wherein the one or more PEG-modified lipids comprise a polyethylene glycol chain of up to 5 kDa in molecular weight covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 length.
5 . The composition according to claim 1 , wherein the lipid carrier vehicle is a liposome or lipid nanoparticle.
6 . The composition according to claim 1 , wherein the one or more mRNAs are encapsulated in the same lipid carrier vehicle.
7 . The composition according to claim 1 , wherein the antibody is comprised of a heavy chain and a light chain, and wherein the heavy chain and the light chain are encoded by a single mRNA.
8 . The composition according to claim 1 , wherein the antibody is comprised of a heavy chain and a light chain, and wherein the heavy chain and the light chain are encoded by separate mRNAs.
9 . The composition according to claim 7 , wherein the heavy chain and the light chain of the antibody are present at varying ratios.
10 . The composition according to claim 1 , wherein the antibody is detectable in a non-lung cell or tissue of a subject for at least 6 hours following administration of the composition to a lung of the subject.
11 . The composition according to claim 1 , wherein the antibody is detectable in a non-lung cell or tissue of a subject for at least 1 day following administration of the composition to a lung of the subject.
12 . The composition according to claim 10 , wherein the non-lung cell is selected from the group consisting of hepatocytes, epithelial cells, hematopoietic cells, epithelial cells, endothelial cells, bone cells, stem cells, mesenchymal cells, neural cells, cardiac cells, adipocytes, vascular smooth muscle cells, cardiomyocytes, skeletal muscle cells, beta cells, pituitary cells, synovial lining cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, macrophages, neutrophils, antigen presenting cells (dendritic cells), fibroblasts, tumor cells and combinations thereof.
13 . The composition according to claim 10 , wherein the non-lung tissue is selected from the group consisting of heart, liver, spleen, kidneys, skeletal muscle, lymph nodes, skin, brain, cerebrospinal fluid, plasma and combinations thereof.
14 . The composition according to claim 1 , wherein the composition is formulated as respirable particles.
15 . The composition according to claim 14 , wherein the respirable particles have a size less than about 500 μm.
16 . The composition according to claim 1 , wherein the composition is formulated as a nebulizable liquid.
17 . The composition according to claim 1 , wherein the composition is formulated as a dry powder.
18 . The composition of claim 1 , wherein the one or more non-cationic lipids is DOPE, the one or more cholesterol-based lipids is Cholesterol, and the one or more PEG-modified lipids is DMG-PEG2000 or DMG-PEG2K.
19 . A composition comprising one or more mRNA encoding an antibody and a lipid carrier vehicle, wherein the composition is formulated for pulmonary delivery, wherein the antibody is a full-length antibody, Fab, Fv, or single-chain Fv, wherein the lipid carrier vehicle comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids, wherein the non-cationic lipid comprises a molar ratio of about 10% to about 70% of the total lipid present in the lipid carrier vehicle.
20 . The composition of claim 1 , wherein the cationic lipid, the non-cationic lipid, Cholesterol, and the PEG-modified lipid are in a molar ratio of 40:30:25:5, 18:56:20:6, or 40:20:35:5, respectively.
21 . The composition of claim 1 , wherein the cationic lipid, DOPE, Cholesterol, and DMG-PEG2000 are in a molar ratio of 40:30:25:5, 18:56:20:6, or 40:20:35:5, respectively.
22 . The composition of claim 1 , wherein the cationic lipid, DSPC, Cholesterol, and DMG-PEG2000 are in a molar ratio of 40:30:25:5, 18:56:20:6, or 40:20:35:5, respectively.
23 . The composition of claim 1 , wherein the device is a metered dose inhaler, a jet-nebulizer, a ultrasonic nebulizer, a dry-powder-inhaler, a propellant-based inhaler, or an insufflator.