STABILIZED ALPHA-GALACTOSIDASE AND USES THEREOF
Multimeric protein structures comprising at least two alpha-galactosidase monomers being covalently linked to one another via a linking moiety are disclosed herein, as well a process for preparing same, and methods of treating Fabry disease via administration of a multimeric protein structure. The disclosed multimeric protein structures exhibit an improved performance, in terms of enhanced activity and/or a longer lasting activity under both lysosomal conditions and in a serum environment.
1 - 20 . (canceled)
21 . A method of treating Fabry disease in a subject in need thereof, said method comprising administering to said subject a stabilized form of α-galactosidase comprising two α-galactosidase monomers covalently linked to one another via a linking moiety, wherein each of said two α-galactosidase monomers has the amino acid sequence of SEQ ID NO: 3,
wherein the stabilized form of α-galactosidase is administered at a dose of about 1 μg/kg to about 500 mg/kg.
22 . The method of claim 21 , wherein the stabilized form of α-galactosidase is administered at a dose of about 1 mg/kg.
23 . The method of claim 21 , wherein the stabilized form of α-galactosidase is administered at a dose of about 2 mg/kg.
24 . The method of claim 21 , wherein the stabilized form of α-galactosidase is administered by an intravenous infusion.
25 . The method of claim 21 , wherein the stabilized form of α-galactosidase is PEGylated.
26 . The method of claim 21 , wherein in the stabilized form of α-galactosidase the linking moiety is a non-peptidic moiety.
27 . The method of claim 26 , wherein the linking moiety comprises poly(alkylene glycol) and at least two functional groups, wherein each functional group forms a covalent bond with one of the native α-galactosidase monomers.
28 . The method of claim 27 , wherein the poly(alkylene glycol) comprises ethylene glycol or propylene glycol units linked together.
29 . The method of claim 26 , wherein the linking moiety has the following general formula:
wherein:
each of X 1 and X 2 is a functional group that forms a covalent bond with at least one α-galactosidase monomer;
C is a carbon atom;
Y is an oxygen atom, a sulfur atom or NR 5 , wherein NR 5 is a nitrogen atom attached to R 5 ;
n is an integer from 5 to 150; and
each of R 1 , R 2 , R 3 , R 4 and R 5 is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, hydroxy, oxo, thiol and thioalkoxy.
30 . The method of claim 29 , wherein at least one of said functional groups forms an amide bond with an α-galactosidase monomer.
31 . The method of claim 29 , wherein said linking moiety is at least 20 atoms long.
32 . The method of claim 29 , wherein n is at least 5.
33 . The method of claim 29 , wherein n is at least 8.
34 . The method of claim 29 , wherein n is not greater than 70.
35 . The method of claim 29 , wherein each of R 1 , R 2 , R 3 , R 4 and R 5 is independently selected from the group consisting of hydrogen and oxo.
36 . The method of claim 21 , wherein the stabilized form of α-galactosidase is glycosylated.
37 . The method of claim 21 , wherein the stabilized form of α-galactosidase is administered as a part of a pharmaceutical composition comprising the stabilized form of α-galactosidase and a pharmaceutically acceptable carrier.
38 . The method of claim 37 , wherein the pharmaceutical composition is an aqueous solution.
39 . The method of claim 37 , wherein the pharmaceutical composition comprises a buffer.
40 . The method of claim 39 , wherein the buffer is a physiological saline buffer.
41 . The method of claim 21 , wherein the subject is a human.