Compositions and methods for treating leber's hereditary optic neuropathy with NADH dehydrogenase proteins
View Patent ↗Disclosed herein is a recombinant nucleic acid, comprising: a mitochondrial targeting sequence; a mitochondrial protein coding sequence, wherein said mitochondrial protein coding sequence encodes a polypeptide comprising a mitochondrial protein; and a 3′UTR nucleic acid sequence. Also disclosed is a pharmaceutical composition comprising the recombinant nucleic acid and a method of treating Leber's hereditary optic neuropathy (LHON) using the pharmaceutical composition.
1. A recombinant nucleic acid, comprising:
a mitochondrial targeting sequence;
a mitochondrial protein coding sequence comprising a sequence that is at least 99% identical to SEQ ID NO: 11 or 12; and
a 3′UTR nucleic acid sequence.
2. The recombinant nucleic acid of claim 1 , wherein said mitochondrial targeting sequence comprises a sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NO: 1-5.
3. The recombinant nucleic acid of claim 1 , wherein said mitochondrial targeting sequence comprises a sequence encodes a polypeptide selected from the group consisting of hsCOX10, hsCOX8, scRPM2, lcSirt5, tbNDUS7, ncQCR2, hsATP5G2, hsLACTB, spilv1, gmCOX2, crATP6, hsOPA1, hsSDHD, hsADCK3, osP0644B06.24-2, Neurospora crassa ATPS (ncATP9), hsGHITM, hsNDUFAB1, hsATP5G3, crATP6_hsADCK3, ncATP9_ncATP9, zmLOC100282174, ncATP9_zmLOC100282174_spilv1_ncATP9, zmLOC100282174_hsADCK3_crATP6_hsATP5G3, zmLOC100282174_hsADCK3_hsATP5G3, ncATP9_zmLOC100282174, hsADCK3_zmLOC100282174_crATP6_hsATP5G3, crATP6_hsADCK3_zmLOC100282174_hsATP5G3, hsADCK3_zmLOC100282174, hsADCK3_zmLOC100282174_crATP6, ncATP9_zmLOC100282174_spilv1_GNFP_ncATP9, and ncATP9_zmLOC100282174_spilv1_lcSirt5_osP0644B06.24-2_hsATP5G2_ncATP9.
4. The recombinant nucleic acid of claim 1 , wherein said mitochondrial targeting sequence encodes a polypeptide comprising a peptide sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NO: 129-159.
5. The recombinant nucleic acid of claim 1 , wherein said 3′UTR nucleic acid sequence comprises a sequence selected from the group consisting of hsACO2, hsATP5B, hsAK2, hsALDH2, hsCOX10, hsUQCRFS1, hsNDUFV1, hsNDUFV2, hsSOD2, hsCOX6c, hsIRP1, hsMRPS12, hsATP5J2, rnSOD2, and hsOXA1L.
6. The recombinant nucleic acid of claim 1 , wherein said 3′UTR nucleic acid sequence comprises a sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NO: 13, 14, and 111-125.
7. The recombinant nucleic acid of claim 1 , wherein said recombinant nucleic acid comprises a sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NO: 25-28, 39-42, 53-56, 67-70, and 81-84.
8. The recombinant nucleic acid of claim 1 , wherein said mitochondrial protein coding sequence encodes a mitochondrial protein comprising or consisting of a sequence that is at least 90% identical to a sequence as set forth in SEQ ID NO: 162.
9. A viral vector comprising said recombinant nucleic acid of claim 1 .
10. The viral vector of claim 9 , wherein said viral vector is an adeno-associated virus (AAV) vector.
11. The viral vector of claim 10 , wherein said AAV vector is a recombinant AAV (rAAV) vector.
12. The viral vector of claim 11 , wherein said rAAV vector is rAAV2 vector.
13. A pharmaceutical composition, comprising a viral vector comprising said recombinant nucleic acid of claim 1 and a pharmaceutically acceptable excipient thereof.
14. The pharmaceutical composition of claim 13 , wherein said viral vector is an adeno-associated virus (AAV) vector.
15. The pharmaceutical composition of claim 13 , wherein said pharmaceutically acceptable excipient comprises phosphate-buffered saline (PBS), α,α-trehalose dehydrate, L-histidine monohydrochloride monohydrate, polysorbate 20, NaCl, NaH 2 PO 4 , Na 2 HPO 4 , KH 2 PO 4 , K 2 HPO 4 , poloxamer 188, or any combination thereof.
16. The pharmaceutical composition of claim 13 , wherein said pharmaceutically acceptable excipient comprises poloxamer 188.
17. The pharmaceutical composition of claim 16 , wherein said pharmaceutically acceptable excipient comprises 0.0001%-0.01% poloxamer 188.
18. The pharmaceutical composition of claim 13 , wherein said pharmaceutical composition has a viral titer of at least 5.0×10 10 vg/mL.
19. The pharmaceutical composition of claim 13 , when said pharmaceutical composition is subject to five freeze/thaw cycles, said pharmaceutical composition retains at least 60% of a viral titer as compared to the viral titer prior to the five freeze/thaw cycles.
20. A method of treating Leber's hereditary optic neuropathy (LHON), comprising intravitreally administering a pharmaceutical composition to a patient in need thereof, wherein said pharmaceutical composition comprises a therapeutically effective amount of an adeno-associated virus (AAV) comprising the recombinant nucleic acid of claim 1 .
21. The method of claim 20 , wherein about 0.01-0.1 mL of said pharmaceutical composition is administered via intravitreal injection.
22. The method of claim 20 , further comprising administering methylprednisolone to said patient.
23. The method of claim 22 , wherein said methylprednisolone is administered daily for at least 2 days prior to intravitreal injection of said pharmaceutical composition.
24. The method of claim 22 , comprising administering methylprednisolone intravenously for at least one day, which is followed by administering methylprednisolone orally for at least a week.
25. The method of claim 22 , wherein said methylprednisolone is administered intravenously at a daily dose of about 80 mg/60 kg.
26. The method of claim 20 , further comprising administering creatine phosphate sodium to said patient.
27. The method of claim 20 , wherein said administering said pharmaceutical composition generates a higher average recovery of vision than a comparable pharmaceutical composition without said recombinant nucleic acid.
28. The method of claim 20 , wherein said administering said pharmaceutical composition generates a higher average recovery of vision than a comparable pharmaceutical composition comprising a recombinant nucleic acid as set forth in SEQ ID NO: 25.
29. A recombinant nucleic acid, comprising a mitochondrial protein coding sequence, wherein said mitochondrial protein coding sequence encodes a polypeptide comprising a mitochondrial protein, wherein said mitochondrial protein coding sequence comprises a sequence that is at least 99% identical to SEQ ID NO: 11 or 12.