Recombinant α-galactosidase A for treatment of Fabry disease
Described are compositions comprising α-galactosidase A enzymes with unique carbohydrate profiles, as well as methods for manufacturing and purifying such enzymes. Also described methods of treating, preventing, and/or ameliorating Fabry Disease by administering such enzymes to a subject in need thereof. Also described are compositions comprising migalastat in combination with such α-galactosidase A enzymes.
1. A human recombinant α-galactosidase A (rhα-Gal A), comprising a protein with an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 2, wherein the protein has less than 10% of total N-linked oligosaccharides that are neutral as measured by normal-phase liquid chromatography on an amino column and the protein has 3.5 to 6 moles of mannose-6-phosphate (M6P) residues per mole of rhα-Gal A homodimer as measured by normal-phase liquid chromatography on an amino column.
2. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has one or more of:
at least 17% of total N-linked oligosaccharides that contain a single sialic acid residue as measured by normal-phase liquid chromatography on an amino column;
at least 20% of total N-linked oligosaccharides that contain two sialic acid residues as measured by normal-phase liquid chromatography on an amino column;
at least 40% of total N-linked oligosaccharides that contain one or two sialic acid residues as measured by normal-phase liquid chromatography on an amino column; or
at least 4 moles of sialic acid residues per mole of rhα-Gal A homodimer as measured by normal-phase liquid chromatography on an amino column.
3. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has at least 50% of total N-linked oligosaccharides that contain sialic acid as measured by normal-phase liquid chromatography on an amino column.
4. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has at least 25% of total N-linked oligosaccharides that are mono-mannose-6-phosphate and at least 6% of total N-linked oligosaccharides that are bis-mannose-6-phosphate as measured by normal-phase liquid chromatography on an amino column.
5. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has at least 7 moles of sialic acid residues per mole of rhα-Gal A homodimer as measured by normal-phase liquid chromatography on an amino column.
6. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has at least 22% of total N-linked oligosaccharides that contain two sialic acid residues as measured by normal-phase liquid chromatography on an amino column.
7. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has at least 14% of total N-linked oligosaccharides that are bis-mannose-6-phosphate as measured by normal-phase liquid chromatography on an amino column.
8. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the rhα-Gal A of claim 1 .
9. The pharmaceutical composition of claim 8 , further comprising a pharmacological chaperone for rhα-Gal A.
10. The pharmaceutical composition of claim 9 , wherein the pharmacological chaperone is migalastat or salt thereof.
11. The pharmaceutical composition of claim 10 , wherein the pharmacological chaperone is migalastat hydrochloride.
12. The pharmaceutical composition of claim 10 , wherein the pharmaceutical composition comprises:
0.5 to 20 μM of the rhα-Gal A; and
50 to 20,000 μM migalastat or salt thereof.
13. The pharmaceutical composition of claim 12 , wherein the pharmacological chaperone is migalastat hydrochloride.
14. The pharmaceutical composition of claim 10 , wherein the pharmaceutical composition comprises:
1 to 10 μM of the rhα-Gal A; and
100 to 10,000 μM migalastat or salt thereof.
15. The pharmaceutical composition of claim 14 , wherein the pharmacological chaperone is migalastat hydrochloride.
16. The pharmaceutical composition of claim 12 , wherein the migalastat or a salt thereof and the rhα-Gal A are present in a molar ratio of the migalastat or a salt thereof to the rhα-Gal A of between 13,000:1 and 50:1.
17. The pharmaceutical composition of claim 16 , wherein the pharmacological chaperone is migalastat hydrochloride.
18. The pharmaceutical composition of claim 8 , wherein the composition is formulated for intravenous, intraarterial, intramuscular, intradermal, subcutaneous, or intraperitoneal administration.
19. A method for treating Fabry disease in a patient, the method comprising administering the pharmaceutical composition of claim 8 to a patient in need thereof.
20. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has 3.5 to 6 moles of mannose-6-phosphate (M6P) residues per mole of rhα-Gal A homodimer and at least 4 moles of sialic acid residues per mole of rhα-Gal A homodimer as measured by normal-phase liquid chromatography on an amino column.
21. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has 4 to 6 moles of mannose-6-phosphate (M6P) residues per mole of rhα-Gal A homodimer and at least 5 moles of sialic acid residues per mole of rhα-Gal A homodimer as measured by normal-phase liquid chromatography on an amino column.
22. The method of claim 19 , further comprising administering migalastat or a salt thereof before, after or simultaneously with the administration of the pharmaceutical composition.
23. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has 1.5% to 5% of total N-linked oligosaccharides that are neutral as measured by normal-phase liquid chromatography on an amino column.
24. The rhα-Gal A of claim 1 , wherein the rhα-Gal A has 3.5 to 4.5 moles of mannose-6-phosphate (M6P) residues per mole of rhα-Gal homodimer as measured by normal-phase liquid chromatography on an amino column.