LIPID NANOPARTICLES CONTAINING POLYNUCLEOTIDES ENCODING GLUCOSE-6-PHOSPHATASE AND USES THEREOF
This disclosure relates to ionizable lipid-based lipid nanoparticles for delivery of mRNA encoding glucose-6-phosphatase. Lipid nanoparticle/mRNA therapies of the invention increase and/or restore deficient levels of glucose-6-phosphatase expression and activity in subjects and are useful for the treatment of glycogen storage disease type 1a (GSD-Ia). Lipid nanoparticle/mRNA therapies of the invention increase glucose production and reduce the abnormal accumulation of glycogen and glucose-6-phosphate associated with GSD-Ia.
1 . A lipid nanoparticle comprising:
(Compound A) or its N-oxide, or a salt or isomer thereof; and
a messenger RNA (mRNA) comprising an open reading frame (ORF) encoding the polypeptide of SEQ ID NO:1, wherein the ORF comprises the nucleic acid sequence of SEQ ID NO:2.
2 . The lipid nanoparticle of claim 1 , wherein the mRNA comprises a 5′ UTR comprising the nucleic acid sequence of SEQ ID NO:55.
3 . The lipid nanoparticle of claim 1 , wherein the mRNA comprises a 3′ UTR comprising the nucleic acid sequence of SEQ ID NO:114.
4 . The lipid nanoparticle of claim 1 , wherein the mRNA comprises the nucleic acid sequence of SEQ ID NO:5.
5 . The lipid nanoparticle of any one of claims 1 to 4 , wherein the mRNA comprises a 5′ terminal cap.
6 . The lipid nanoparticle of claim 5 , wherein the 5′ terminal cap comprises a guanine cap nucleotide containing an N 7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl.
7 . The lipid nanoparticle of any one of claims 1 to 6 , wherein the mRNA comprises a poly-A region.
8 . The lipid nanoparticle of claim 7 , wherein the poly-A region is at least about 100 nucleotides in length.
9 . The lipid nanoparticle of any one of claims 1 to 8 , wherein all of the uracils of the mRNA are N 1 -methylpseudouracils.
10 . The lipid nanoparticle of claim 1 , wherein the mRNA comprises a 5′ terminal cap comprising a guanine cap nucleotide containing an N 7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl, wherein the mRNA comprises the nucleotide sequence of SEQ ID NO:5, wherein the mRNA comprises a poly-A region at least about 100 nucleotides in length, and wherein all of the uracils of the mRNA are N 1 -methylpseudouracils.
11 . The lipid nanoparticle of any one of claims 1 to 10 , wherein the lipid nanoparticle comprises Compound I.
12 . The lipid nanoparticle of any one of claims 1 to 11 , wherein the lipid nanoparticle comprises a phospholipid and a structural lipid.
13 . The lipid nanoparticle of any one of claims 1 to 10 , wherein the lipid nanoparticle comprises: (a) cholesterol and Compound I; (b) cholesterol and PEG-DMG; (c) DSPC, cholesterol, and Compound I; (d) DOPE, cholesterol, and Compound I; (e) DSPC, cholesterol, and PEG-DMG; or (f) DOPE, cholesterol, and PEG-DMG.
14 . A method of expressing a glucose-6-phosphatase (G6PC) polypeptide in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
15 . A method of treating, preventing, or delaying the onset and/or progression of glycogen storage disease type 1a (GSD-Ia) in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
16 . A method of increasing blood, plasma, and/or serum glucose levels in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
17 . A method of reducing liver glycogen levels in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
18 . A method of reducing liver glucose-6-phosphate (G6P) levels in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
19 . A method of reducing serum and/or liver triglyceride levels in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
20 . A method of increasing G6PC activity in a human subject in need thereof, comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
21 . The method of claim 20 , wherein the G6PC activity is increased in the liver and/or blood of the human subject.
22 . A method of treating or preventing liver adenoma in a human subject having glycogen storage disease type 1a (GSD-Ia), comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
23 . A method of treating or preventing liver carcinoma in a human subject having glycogen storage disease type 1a (GSD-Ia), comprising administering to the human subject an effective amount of the lipid nanoparticle of any one of claims 1 to 13 .
24 . The method of any one of claims 14 to 23 , wherein the human subject has been fasting.
25 . The method of claim 24 , wherein the human subject has been fasting for at least 2 hours.
26 . The method of any one of claims 14 to 25 , comprising multiple administrations of the lipid nanoparticle to the human subject.
27 . The method of claim 26 , wherein the multiple administrations to the human subject are about once a week, about once every two weeks, or about once a month.
28 . The method of any one of claims 14 to 27 , wherein the mRNA is administered at a dose of 0.1 mg mRNA/kg of body weight of the human subject.
29 . The method of any one of claims 14 to 27 , wherein the mRNA is administered at a dose of 0.2 mg mRNA/kg of body weight of the human subject.
30 . The method of any one of claims 14 to 27 , wherein the mRNA is administered at a dose of 0.5 mg mRNA/kg of body weight of the human subject.
31 . The method of any one of claims 14 to 30 , wherein the lipid nanoparticle is administered intravenously.