Human alpha-galactosidase variants
The present invention provides engineered human alpha-galactosidase polypeptides and compositions thereof. The engineered human alpha-galactosidase polypeptides have been optimized to provide improved stability under both acidic (pH<4.5) and basic (pH>7) conditions. The invention also relates to the use of the compositions comprising the engineered human alpha-galactosidase polypeptides for therapeutic purposes.
1. A recombinant alpha-galactosidase A comprising a polypeptide sequence comprising at least 90% sequence identity to SEQ ID NO: 46, wherein the polypeptide sequence comprises mutations at 316D, 10P, 39E, 44R, 47T, 92H 166P, 206A, 217R, 247D, 261G, 271A, 302K, 3221, 337P, 368A, and 392T.
2. The recombinant alpha galactosidase A of claim 1 , wherein said alpha galactosidase A is SEQ ID NO: 46.
3. The recombinant alpha galactosidase A of claim 1 , wherein said recombinant alpha galactosidase A is derived from a human alpha galactosidase A.
4. The recombinant alpha galactosidase A of claim 1 , wherein said recombinant alpha galactosidase A is a deimmunized alpha galactosidase A.
5. The recombinant alpha galactosidase A of claim 1 , wherein said recombinant alpha galactosidase A exhibits at least one improved property selected from: i) enhanced catalytic activity; ii) increased tolerance to pH 7.4; iii) increased tolerance to pH 4.3; iv) increased tolerance to serum; v) reduced immunogenicity; or vi) enhanced thermostability; or a combination of any of i), ii), iii), iv), v), or vi), as compared to SEQ ID NO: 5.
6. A composition comprising at least one recombinant alpha galactosidase A of claim 1 .
7. A recombinant polynucleotide sequence encoding at least one recombinant alpha galactosidase A of claim 1 , or optionally said polynucleotide sequence is codon optimized.
8. An expression vector comprising the recombinant polynucleotide sequence of claim 7 .
9. The expression vector of claim 8 , wherein said recombinant polynucleotide sequence is operably linked to a control sequence, or optionally, wherein the control sequence is a promoter, or, wherein the promoter is a heterologous promoter.
10. A host cell comprising the expression vector of claim 7 .
11. A method of producing an alpha galactosidase A variant, comprising culturing said host cell of claim 10 , under conditions that said alpha galactosidase A encoded by said recombinant polynucleotide is produced.
12. The method of claim 11 , further comprising recovering said alpha galactosidase A, or optionally purifying said alpha galactosidase A.
13. A pharmaceutical composition for the treatment of Fabry disease, comprising the composition of claim 6 and a pharmaceutically acceptable carrier and/or excipient.
14. The pharmaceutical composition of claim 13 , wherein said composition is suitable for parenteral injection or infusion to a human.
15. A method for treating the symptoms of Fabry disease in a subject, comprising providing a subject having Fabry disease, with the pharmaceutical composition of claim 13 .
16. The method of claim 15 , wherein said symptoms of Fabry disease are ameliorated.
17. The method of claim 16 , wherein said subject is able to eat a diet that is less restricted in its fat content than diets required by subjects exhibiting the symptoms of Fabry disease.
18. The method of claim 15 , wherein said subject is an infant or a child.
19. The method of claim 15 , wherein said subject is an adult or a young adult.