Granted Patent B2

US 12,385,026 · App. 17/716,348 · Granted Aug 12, 2025

Inventors: Avidor Shulman (Rakefet, IL); Ilya Ruderfer (Carmiel, IL); Tehila Ben-Moshe (Koranit, IL); Talia Shekhter (Petach-Tikva, IL); Yaniv Azulay (Akko, IL); Tali Kizhner (Atzmon-Segev, IL); Yoseph Shaaltiel (Timrat, IL)

Assignee: Protalix Ltd.

C12N9/2465C12Y302/01022A61K38/00

Abstract

Claims (32)

1. 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.

2. The stabilized form of α-galactosidase of claim 1 , wherein the linking moiety is a non-peptidic moiety.

3. The stabilized form of α-galactosidase of claim 2 , 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.

4. The stabilized form of α-galactosidase of claim 3 , wherein the poly (alkylene glycol) comprises ethylene glycol or propylene glycol units linked together.

5. The stabilized form of α-galactosidase of claim 3 , wherein the linking moiety has the following general formula:

—X 1 —(CR 1 R 2 —CR 3 R 4 —Y) n —X 2 —

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.

6. The stabilized form of α-galactosidase of claim 5 , wherein at least one of said functional groups forms an amide bond with an α-galactosidase monomer.

7. The stabilized form of α-galactosidase of claim 5 , wherein said linking moiety is at least 20 atoms long.

8. The stabilized form of α-galactosidase of claim 5 , wherein n is at least 25.

9. The stabilized form of α-galactosidase of claim 5 , wherein n is an integer from 40 to 70.

10. The stabilized form of α-galactosidase of claim 5 , 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.

11. The stabilized form of α-galactosidase of claim 1 , featuring a characteristic selected from the group consisting of:

(a) an α-galactosidase activity upon subjecting the stabilized form of α-galactosidase to human plasma conditions for one hour which is at least 10% higher than an activity of naturally occurring α-galactosidase upon subjecting said naturally occurring α-galactosidase to said human plasma conditions for one hour;

(b) an α-galactosidase activity which decreases upon subjecting the stabilized form of α-galactosidase to human plasma conditions for one hour by a percentage which is at least 10% less than the percentage by which an activity of naturally occurring α-galactosidase decreases upon subjecting said naturally occurring α-galactosidase to said human plasma conditions for one hour;

(c) an α-galactosidase activity which remains in a range of 50% to 150% of the initial activity upon subjecting the stabilized form of α-galactosidase to human plasma conditions for one hour;

(d) an α-galactosidase activity upon subjecting the stabilized form of α-galactosidase to lysosomal conditions for one week, which is at least 10% higher than an activity of naturally occurring α-galactosidase upon subjecting said naturally occurring α-galactosidase to said lysosomal conditions for one week;

(e) an α-galactosidase activity which decreases upon subjecting the stabilized form of α-galactosidase to lysosomal conditions for one day by a percentage which is at least 10% less than the percentage by which an activity of naturally occurring α-galactosidase decreases upon subjecting said naturally occurring α-galactosidase to said lysosomal conditions for one day;

(f) an α-galactosidase activity which remains in a range of 50% to 150% of the initial activity upon subjecting the stabilized form of α-galactosidase to lysosomal conditions for one day;

(g) an α-galactosidase activity immediately upon subjecting the stabilized form of α-galactosidase to lysosomal conditions, which is at least 10% higher than an activity of naturally occurring α-galactosidase immediately upon subjecting said naturally occurring α-galactosidase to said lysosomal conditions;

(h) an α-galactosidase activity immediately upon subjecting the stabilized form of α-galactosidase to an aqueous solution having a pH of 7 and a temperature of 37° C., which is at least 10% higher than an activity of naturally occurring α-galactosidase immediately upon subjecting said naturally occurring α-galactosidase to said aqueous solution having a pH of 7 and a temperature of 37° C.; and

(i) a circulating half-life in human plasma which is higher than a circulating half-life of naturally occurring α-galactosidase.

12. A pharmaceutical composition comprising the stabilized form of α-galactosidase of claim 11 and a pharmaceutically acceptable carrier.

13. A method of treating Fabry disease in a subject in need thereof, said method comprising administering to said subject the stabilized form of α-galactosidase of claim 1 , such that said Fabry disease in said subject is treated.

14. The method of claim 13 , wherein said administering is by an intravenous infusion.

15. A process of preparing the stabilized form of α-galactosidase of claim 1 , said process comprising reacting α-galactosidase comprising two α-galactosidase monomers with a cross-linking agent, wherein said cross-linking agent comprises a linking moiety and at least two reactive groups.

16. The process of claim 15 , wherein said reactive groups comprise a leaving group.

Assignments (1)

Continuity (12)

References Cited (259)