POLYNUCLEOTIDES ENCODING METHYLMALONYL-COA MUTASE FOR THE TREATMENT OF METHYLMALONIC ACIDEMIA
This disclosure relates to mRNA therapy for the treatment of methylmalonic acidemia (MMA). mRNAs for use in the invention, when administered in vivo, encode methylmalonyl-CoA mutase (MUT). mRNA therapies of the disclosure increase and/or restore deficient levels of MUT expression and/or activity in subjects.
1 . A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject by intravenous infusion a lipid nanoparticle comprising an open reading frame (ORF) encoding the human methylmalonyl-CoA mutase (MUT) polypeptide of SEQ ID NO:1, wherein the ORF is at least 96% identical to the nucleotide sequence of SEQ ID NO:7, and wherein the mRNA is administered at a dose of 0.01 mg/kg to 2.0 mg/kg.
2 . The method of claim 1 , wherein the ORF is at least 97% identical to the nucleotide sequence of SEQ ID NO:7.
3 . The method of claim 1 , wherein the ORF is at least 98% identical to the nucleotide sequence of SEQ ID NO:7.
4 . The method of claim 1 , wherein the ORF is at least 99% identical to the nucleotide sequence of SEQ ID NO:7.
5 . The method of claim 1 , wherein the ORF is 100% identical to the nucleotide sequence of SEQ ID NO:7.
6 . The method of any one of claims 1-5 , wherein the mRNA comprises a 5′ UTR comprising the nucleotide sequence of SEQ ID NO:78.
7 . The method of any one of claims 1-6 , wherein the mRNA comprises a 3′ UTR comprising the nucleotide sequence of SEQ ID NO:136.
8 . The method of claim 1 , wherein the mRNA comprises the nucleic acid sequence of SEQ ID NO:10.
9 . The method of any one of claims 1-8 , wherein the mRNA comprises a 5′ terminal cap.
10 . The method of claim 9 , wherein the 5′ terminal cap comprises a guanine cap nucleotide containing an N7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl.
11 . The method of any one of claims 1 to 10 , wherein the mRNA comprises a poly-A region.
12 . The method of claim 11 , wherein the mRNA comprises a poly-A tail 100 residues in length.
13 . The method of any one of claims 1-12 , wherein all of the uracils of the mRNA are N1-methylpseudouracils.
14 . The method of claim 1 , wherein the mRNA comprises a 5′ terminal cap comprising a guanine cap nucleotide containing an N7 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:10, 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 N1-methylpseudouracils.
15 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of 0.1 mg/kg to 0.6 mg/kg.
16 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of about 0.1 mg/kg.
17 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of about 0.2 mg/kg.
18 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of about 0.3 mg/kg.
19 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of about 0.4 mg/kg.
20 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of about 0.5 mg/kg.
21 . The method of any one of claims 1 to 14 , wherein the mRNA is administered at a dose of about 0.6 mg/kg.
22 . The method of any one of claims 1 to 21 , wherein the lipid nanoparticle is administered at intervals of about once every 2 weeks.
23 . The method of any one of claims 1 to 21 , wherein the lipid nanoparticle is administered at intervals of about once every 3 weeks.
24 . The method of any one of claims 1 to 21 , wherein the lipid nanoparticle is administered at intervals of about once every 4 weeks.
25 . The method of any one of claims 1 to 24 , comprising administering at least 12 doses of the lipid nanoparticle.
26 . The method of any one of claims 1 to 25 , wherein the human subject is ≥1 to ≤18 years of age.
27 . The method of any one of claims 1 to 25 , wherein the human subject is ≥1 year of age to <2 years of age.
28 . The method of any one of claims 1 to 25 , wherein the human subject is ≥2 years of age to <12 years of age.
29 . The method of any one of claims 1 to 25 , wherein the human subject is ≥12 years of age to ≤18 years of age.
30 . The method of any one of claims 1 to 29 , wherein the human subject is administered at least one of an H 2 blocker, an H 1 blocker, or acetaminophen/paracetamol prior to infusion of the lipid nanoparticle.
31 . The method of any one of claims 1 to 29 , wherein the human subject is administered an H 2 blocker, an H 1 blocker, and acetaminophen/paracetamol prior to infusion of the lipid nanoparticle.
32 . The method of any one of claims 1 to 31 , wherein the methylmalonic academia is isolated methylmalonic acidemia due to methylmalonyl-CoA mutase deficiency.
33 . The method of any one of claims 1 to 32 , wherein the treatment reduces methylmalonic acid levels from baseline.
34 . The method of any one of claims 1 to 32 , wherein the treatment reduces 2-methylcitric acid levels from baseline.
35 . The method of any one of claims 1 to 32 , wherein the treatment reduces methylmalonic acid and 2-methylcitric acid levels from baseline.
36 . The method of any one of claims 1 to 32 , wherein the treatment increases MUT mRNA levels from baseline.
37 . The method of any one of claims 1 to 32 , wherein the treatment reduces the frequency and duration of clinically significant events.
38 . The method of any one of claims 1 to 32 , wherein the treatment reduces the frequency and duration of metabolic decompensation events.
39 . The method of any one of claims 1 to 32 , wherein the treatment reduces the incidence and duration of healthcare utilization visits.
40 . The method of any one of claims 1 to 32 , wherein the treatment increases Pediatric Quality-of-Life Inventory measurements.
41 . The method of any one of claims 1 to 32 , wherein the treatment increases height and weight growth velocity of the human subject.
42 . The method of any one of claims 1 to 41 , wherein the lipid nanoparticle comprises a compound of Formula (I):
or its N-oxide, or a salt or isomer thereof,
wherein R′ a is R′ branched ; wherein
R′ branched is:
wherein
denotes a point of attachment;
wherein R aα , R aβ , R aγ , and R aδ are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl;
R 2 and R 3 are each independently selected from the group consisting of C 1-14 alkyl and C 2-14 alkenyl;
R 4 is selected from the group consisting of —(CH 2 ) n OH, wherein n is selected from the group consisting of 1, 2, 3, 4, and 5, and
wherein
denotes a point of attachment; wherein
R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
each R 5 is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H;
each R 6 is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H;
M and M′ are each independently selected from the group consisting of —C(O)O— and —OC(O)—;
R′ is a C 1-12 alkyl or C 2-12 alkenyl;
l is selected from the group consisting of 1, 2, 3, 4, and 5; and
m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13.
43 . The method of claim 42 , wherein the lipid nanoparticle further comprises a phospholipid, a structural lipid, and a PEG-lipid.
44 . The method of claim 43 , wherein the PEG-lipid is Compound I.
45 . The method of claim 43 or 44 , wherein the lipid nanoparticle comprises:
(i) 40-50 mol % of the compound of Formula (I), 30-45 mol % of the structural lipid, 5-15 mol % of the phospholipid, and 1-5 mol % of the PEG-lipid; or
(ii) 45-50 mol % of the compound of Formula (I), 35-45 mol % of the structural lipid, 8-12 mol % of the phospholipid, and 1.5 to 3.5 mol % of the PEG-lipid.
46 . The method of any one of claims 1 to 41 , wherein the lipid nanoparticle comprises:
(i) Compound II, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;
(i) Compound VI, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;
(i) Compound II, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;
(i) Compound VI, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;
(i) Compound II, (ii) Cholesterol, and (iii) Compound I;
(i) Compound II, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) Compound I;
(i) Compound B, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;
(i) Compound B, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;
(i) Compound B, (ii) Cholesterol, and (iii) Compound I;
(i) Compound B, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) Compound I;
(i) Compound A, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;
(i) Compound A, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;
(i) Compound A, (ii) Cholesterol, and (iii) Compound I; or
(i) Compound A, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) Compound I.
47 . The method of any one of claims 1 to 41 , wherein the lipid nanoparticle comprises Compound II and Compound I.
48 . The method of any one of claims 1 to 41 , wherein the lipid nanoparticle comprises Compound B and Compound I.
49 . The method of any one of claims 1 to 41 , wherein the lipid nanoparticle comprises Compound A and Compound I.
50 . The method of any one of claims 1 to 41 , wherein the lipid nanoparticle comprises Compound II, DSPC, Cholesterol, and Compound I.