Polynucleotides encoding methylmalonyl-CoA mutase
The disclosure relates to polynucleotides comprising an open reading frame of linked nucleosides encoding human methylmalonyl-CoA mutase precursor, human methylmalonyl-CoA mutase (MCM) mature form, or functional fragments thereof. In some embodiments, the disclosure includes methods of treating methylmalonic acidemia in a subject in need thereof comprising administering an mRNA encoding an MCM polypeptide.
1. A lipid nanoparticle composition comprising:
a messenger RNA (mRNA) that comprises an open reading frame (ORF) encoding a methylmalonyl-CoA mutase (MCM) polypeptide, and
a compound of Formula (IA):
or a salt or stereoisomer thereof, wherein:
l is selected from 1, 2, 3, 4, and 5;
m is selected from 5, 6, 7, 8, and 9;
M 1 is a bond or M′;
R 4 is unsubstituted C 1-3 alkyl, or —(CH 2 ) n Q, wherein n is 1, 2, 3, 4 or 5 and Q is OH, —NHC(S)N(R) 2 , or —NHC(O)N(R) 2 ;
M and M′ are independently selected from —C(O)O, OC(O), C(O)N(R′), P(O)(OR′)O, an aryl group, and a heteroaryl group;
R 2 and R 3 are independently selected from the group consisting of H, C 1-14 alkyl, and C 2-14 alkenyl;
each R is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H;
each R′ is independently selected from the group consisting of C 1-18 alkyl, C 2-18 alkenyl, —R*YR″, —YR″, and H;
each R″ is independently selected from the group consisting of C 3-14 alkyl and C 3-14 alkenyl;
each R* is independently selected from the group consisting of C 1-12 alkyl and C 2-12 alkenyl; and
each Y is independently a C 3-6 carbocycle.
2. The lipid nanoparticle composition of claim 1 , wherein the compound is a compound of Formula (II):
or a salt or stereoisomer thereof, wherein:
R 4 is unsubstituted C 1-3 alkyl, or —(CH 2 ) n Q, wherein n is 2, 3, or 4 and Q is OH, —NHC(S)N(R) 2 , or —NHC(O)N(R) 2 .
3. The lipid nanoparticle composition of claim 1 , wherein the compound is a compound of Formula (IIa), (IIb), (IIc), or (IIe):
or a salt or stereoisomer thereof.
4. The lipid nanoparticle composition of claim 1 , wherein M and M′ are independently —C(O)O— or —OC(O)—.
5. The lipid nanoparticle composition of claim 1 , wherein R 2 and R 3 are the same.
6. The lipid nanoparticle composition of claim 1 , wherein R 2 and R 3 are C 8 alkyl.
7. The lipid nanoparticle composition of claim 1 , wherein R 2 and R 3 are different.
8. The lipid nanoparticle composition of claim 1 , wherein n is 2.
9. The lipid nanoparticle composition of claim 1 , wherein R′ is selected from: C 4 alkyl, C 4 alkenyl, C 5 alkyl, C 5 alkenyl, C 6 alkyl, C 6 alkenyl, C 7 alkyl, C 7 alkenyl, C 9 alkyl, C 9 alkenyl, C 11 alkyl, C 11 alkenyl, C 12 alkyl, C 12 alkenyl, C 13 alkyl, C 13 alkenyl, C 14 alkyl, C 14 alkenyl, C 15 alkyl, C 15 alkenyl, C 16 alkyl, C 16 alkenyl, C 17 alkyl, C 17 alkenyl, C 18 alkyl, and C 18 alkenyl.
10. The lipid nanoparticle composition of claim 1 , wherein l is selected from 3, 4, and 5.
11. The lipid nanoparticle composition of claim 1 , wherein:
(i) n is 2;
(ii) R′ is selected from: C 4 alkyl, C 4 alkenyl, C 5 alkyl, C 5 alkenyl, C 6 alkyl, C 6 alkenyl, C 7 alkyl, C 7 alkenyl, C 9 alkyl, C 9 alkenyl, C 11 alkyl, C 11 alkenyl, C 12 alkyl, C 12 alkenyl, C 13 alkyl, C 13 alkenyl, C 14 alkyl, C 14 alkenyl, C 15 alkyl, C 15 alkenyl, C 16 alkyl, C 16 alkenyl, C 17 alkyl, C 17 alkenyl, C 18 alkyl, and C 18 alkenyl; and
(iii) l is selected from 3, 4, and 5.
12. The lipid nanoparticle composition of claim 1 , wherein the compound is a compound of formula:
or a salt thereof.
13. The lipid nanoparticle composition of claim 1 , further comprising a phospholipid, a structural lipid, and a PEG lipid.
14. The lipid nanoparticle composition of claim 13 , wherein:
(i) the phospholipid is selected from the group consisting of:
1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC),
1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC),
1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC),
1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC),
1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC),
1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC),
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC),
1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC),
1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC),
1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC),
1,2-dilinolenoyl-sn-glycero-3-phosphocholine,
1,2-diarachidonoyl-sn-glycero-3-phosphocholine,
1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine,
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE),
1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16:0 PE),
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine,
1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine,
1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine,
1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine,
1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG),
sphingomyelin, and
any mixtures thereof;
(ii) the structural lipid is cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, or any mixture thereof; and
(iii) the PEG lipid is a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, or any mixture thereof.
15. The lipid nanoparticle composition of claim 13 , wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.
16. The lipid nanoparticle composition of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, or SEQ ID NO:213.
17. The lipid nanoparticle composition of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:213.
18. The lipid nanoparticle composition of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:208.
19. The lipid nanoparticle composition of claim 12 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:213.