MESSENGER RNA THERAPY FOR TREATMENT OF ARTICULAR DISEASE
The present invention provides, among other things, a method of intra-articular delivery of messenger RNA (mRNA), comprising administering into a joint of a subject in need of delivery a composition comprising an mRNA encoding a protein, such that the administering of the composition results in expression of the protein encoded by the mRNA in the joint.
1 - 46 . (canceled)
47 . A pharmaceutical composition comprising one or more mRNAs encoding one or more polypeptides for treating a joint disease, disorder or condition,
wherein the one or more mRNAs encoding the one or more polypeptides encode an antibody, a soluble receptor, one or more polypeptides deficient in the joint disease, disorder, or condition, or a wild-type or naturally occurring protein sequence that is normally present in a healthy joint,
wherein the one or more mRNAs are encapsulated within one or more nanoparticles, and
wherein the composition is formulated for intra-articular delivery.
48 . The composition according to claim 47 , wherein the expression and/or activity of the one or more polypeptides is: (a) detected in cartilage, ligament, muscle, synovial intimal cells, chondrocytes and/or fluids of the joint; and/or (b) detectable at least about 6 hours, 12 hours, 18 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or 1 month post-administration.
49 . The composition according to claim 47 , wherein the one or more mRNAs are unmodified.
50 . The composition according to claim 47 , wherein the one or more mRNAs are modified.
51 . The composition according to claim 47 , wherein the composition comprises two distinct mRNAs encoding an antibody heavy chain and an antibody light chain, respectively.
52 . The composition according to claim 47 , wherein the antibody encoded by the one or more mRNAs is an anti-TNFα antibody, anti-IL-6 antibody, anti-IL-1β antibody, anti-sTNFR-I antibody, anti-sTNFR-II antibody, anti-IL-2 antibody, anti-IFN-α antibody, anti-IL-18 antibody, anti-IL-12 antibody, or anti-IL-12p40 antibody.
53 . The composition according to claim 47 , wherein the antibody is an intact immunoglobulin, (Fab) 2 , (Fab′) 2 , Fab, Fab′ or scFv.
54 . The composition according to claim 47 , wherein the one or more nanoparticles comprise a lipid nanoparticle or a liposome.
55 . The composition according to claim 47 , wherein the one or more nanoparticles comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.
56 . The composition according to claim 55 , wherein the one or more cationic lipids are selected from the group consisting of C12-200, MC3, DLinDMA, DLinkC2DMA, cKK-E12, ICE (Imidazol-based), HGT5000, HGT5001, DODAC, DDAB, DMRIE, DOSPA, DOGS, DODAP, DODMA and DMDMA, DODAC, DLenDMA, DMRIE, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DLinDAP, DLincarbDAP, DLinCDAP, KLin-K-DMA, DLin-K-XTC2-DMA, HGT4003, and combinations thereof.
57 . The composition according to claim 55 , wherein the one or more non-cationic lipids are selected from DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-dioleyl-sn-glycero-3-phosphotidylcholine) DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)) and combinations thereof.
58 . The composition according to claim 55 , wherein the one or more cholesterol-based lipids is cholesterol or PEGylated cholesterol.
59 . The composition according to claim 58 , wherein the one or more PEG-modified lipids comprise a poly (ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 length.
60 . The composition according to claim 55 , wherein:
(a) cKK-E12, DOPE, cholesterol, and DMG-PEG2K are at a ratio of about 50:25:20:5, 50:20:25:5, 50:27:20:3, 40:30:20:10, 40:30:25:5, or 40:32:25:3; or
(b) C12-200, DOPE, cholesterol and DMG-PEG2K are at a ratio of about 50:25:20:5, 50:20:25:5, 50:27:20:3, 40:30:20:10, 40:30:25:5, or 40:32:25:3; or
(c) HGT5001, DOPE, cholesterol and DMG-PEG2K are at a ratio of about 50:25:20:5, 50:20:25:5, 50:27:20:3, 40:30:20:10, 40:30:25:5, or 40:32:25:3.
61 . The composition according to claim 47 , wherein the one or more nanoparticles have a size less than about 40-100 nm.
62 . The composition according to claim 47 , wherein each of the one or more mRNAs has a length of or greater than about 0.5 kb, 1 kb, 1. 5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, 5 kb, 6kb, 7 kb, 8 kb, 9 kb, 10 kb, 11 kb, 12 kb, 13 kb, 14 kb, or 15 kb.
63 . The composition according to claim 50 , wherein the one or more modified mRNAs comprise one or more modified nucleotides, wherein the one or more modified nucleotides comprise pseudouridine, N-1-methyl-pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and/or 2-thiocytidine.
64 . The composition according to claim 50 , wherein the one or more modified mRNAs comprise a backbone modification selected from the group consisting of a modification of the phosphates of the backbone into methylphosphonates, methylphosphoramidates, phosphoramidates, phosphorothioates, cytidine 5′-O-(1-thiophosphate), boranophosphates, and positively charged guanidinium groups.
65 . The composition according to claim 50 , wherein the one or more modified mRNAs comprise a backbone modification selected from the group consisting of a sugar modification selected from the group consisting of 2′-deoxy-2′-fluoro-oligoribonucleotide (2′-fluoro-2′-deoxycytidine 5′-triphosphate, 2′-fluoro-2′-deoxyuridine 5′-triphosphate), 2′-deoxy-2′-deamine-oligoribonucleotide (2′-amino-2′-deoxycytidine 5′-triphosphate, 2′-amino-2′-deoxyuridine 5′-triphosphate), 2′-O-alkyloligoribonucleotide, 2′-deoxy-2′-C-alkyloligoribonucleotide (2′-O-methylcytidine 5′-triphosphate, 2′-methyluridine 5′-triphosphate), 2′-C-alkyloligoribonucleotide, and isomers thereof (2′-aracytidine 5′-triphosphate, 2′-arauridine 5′-triphosphate), and azidotriphosphates (2′-azido-2′-deoxycytidine 5′-triphosphate, 2′-azido-2′-deoxyuridine 5′-triphosphate).
66 . The composition according to claim 50 , wherein the one or more modified mRNAs comprise a base modification selected from the group consisting of 2-amino-6-chloropurine riboside 5′-triphosphate, 2-aminoadenosine 5′-triphosphate, 2-thiocytidine 5′-triphosphate, 2-thiouridine 5′-triphosphate, 4-thiouridine 5′-triphosphate, 5-aminoallylcytidine 5′-triphosphate, 5-aminoallyluridine 5′-triphosphate, 5-bromocytidine 5′-triphosphate, 5-bromouridine 5′-triphosphate, 5-iodocytidine 5′-triphosphate, 5-iodouridine 5′-triphosphate, 5-methylcytidine 5′-triphosphate, 5-methyluridine 5′-triphosphate, 6-azacytidine 5′-triphosphate, 6-azauridine 5′-triphosphate, 6-chloropurine riboside 5′-triphosphate, 7-deazaadenosine 5′-triphosphate, 7-deazaguanosine 5′-triphosphate, 8-azaadenosine 5′-triphosphate, 8-azidoadenosine 5′-triphosphate, benzimidazole riboside 5′-triphosphate, N1-methyladenosine 5′-triphosphate, N1-methylguanosine 5′-triphosphate, N6-methyladenosine 5′-triphosphate, O6-methylguanosine 5′-triphosphate, pseudouridine 5′-triphosphate, puromycin 5′-triphosphate and xanthosine 5′-triphosphate.