Stabilized compositions of proteins having a free thiol moiety

Patent metadata

Field	Value
Publication number	US-9694057-B2
Application number	US-201514743427-A
Country	US
Kind code	B2
Filing date	Jun 18, 2015
Priority date	Feb 7, 2006
Publication date	Jul 4, 2017
Grant date	Jul 4, 2017

Abstract

Compositions of proteins having free thiols, and methods of making and using such compositions, are described.

First claim

What is claimed is:

1. A method of producing a composition comprising glucocerebrosidase (GCB), the method comprising providing a GCB preparation and formulating the GCB with a carbohydrate, wherein the carbohydrate is present in an amount sufficient to maintain the stability of the GCB, wherein the carbohydrate is sucrose or trehalose, and wherein the pH of the composition is less than 7.0.

2. The method of claim 1 , wherein the stability is at least 5-80% greater, under pre-selected conditions, than the stability of a composition which differs by lacking the carbohydrate.

3. The method of claim 1 , wherein the carbohydrate is present in an amount sufficient to increase the stability of the GCB.

4. The method of claim 1 , wherein the carbohydrate is present in an amount sufficient to inhibit the reaction of a free thiol on a first molecule of the GCB with a free thiol on a second molecule of the GCB to form an aggregate.

5. The method of claim 1 , wherein the carbohydrate is present in an amount sufficient to inhibit the formation of an aggregate formed by the reaction of a free thiol on a first molecule of the GCB with a free thiol on a second molecule of the GCB by at least 5-80%, under pre-selected conditions, as compared to the same composition lacking the carbohydrate.

6. The method of claim 1 , wherein the carbohydrate is present in an amount sufficient that upon storage, in a gas tight container, at a temperature of 2-8° C., for a period of months, the composition will retain at least 85% of the stability the composition had prior to storage.

7. The method of claim 6 , wherein the storage occurs in darkness.

8. The method of claim 1 , wherein the carbohydrate is present in an amount sufficient to have stability comparable to that of a lyophilized composition comprising sucrose, 0.01% polysorbate-20, pH 6.0, 50 mM Citrate.

9. The method of claim 1 , wherein the carbohydrate is present in an amount sufficient to maintain biochemical integrity and bioactivity characteristics of the GCB.

10. The method of claim 1 , wherein the pH of the composition is between about 4.5 and about 6.5.

11. The method of claim 1 , wherein the composition further comprises an antioxidant, wherein the antioxidant and the carbohydrate are present in amounts sufficient to maintain the stability of the GCB.

12. The composition of claim 11 , wherein the antioxidant is cysteine, cysteine-HCl, or methionine.

13. The method of claim 1 , wherein the composition further comprises a surfactant.

14. The method of claim 13 , wherein the surfactant is poloxamer 188.

15. The method of claim 1 , wherein the composition contains less than about 10% of O 2 .

16. The method of claim 1 , further comprising physical removal of O 2 from the composition.

17. The method of claim 1 , wherein the GCB has two, three, or more free thiol groups and has zero, two, four, or more thiol groups which form sulfhydryl bridges, per active unit of GCB.

18. The method of claim 1 , further comprising exposing the composition to an inert gas, wherein the inert gas is present in a concentration higher than in the ambient atmosphere.

19. The method of claim 1 , further comprising packaging the composition.

20. The method of claim 19 , wherein the packaging comprises contacting the GCB with an inert gas to reduce the amount of a reactive species, and introducing the GCB, the inert gas, and the carbohydrate into a gas tight container.

21. The method of claim 20 , wherein the container comprises a headspace comprising at least 90% (vol/vol) an inert gas.

22. The method of claim 21 , wherein the container is a prefilled syringe, a vial, or ampoule.

23. The method of claim 22 , wherein the prefilled syringe is a needleless syringe.

24. The method of claim 20 , wherein the inert gas is N 2 or Ar and the reactive species is O 2 .

25. The method of claim 1 , wherein the composition is a liquid.

26. The method of claim 22 , wherein carbohydrate is present at between about 1 and about 40% (wt/vol).

27. The method of claim 22 , wherein the antioxidant is present at between about 0.001 and about 10% (wt/vol).

28. The method of claim 22 , wherein the composition comprises about 0.1-40 mg/ml GCB, about 0.001-10% cysteine, about 1-40% sucrose, at a pH of about 5.5-6.0, and wherein the level of dissolved O 2 is less than about 10%.

29. The method of claim 1 , wherein the composition does not comprise polysorbate.

Assignees

• Shire Human Genetic Therapies
• Shire Huma Genetic Therapies Inc

Inventors

• Zhu Gaozhong
• Nguyen Vinh
• Lowe Kris
• Shahrokh Zahra

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title

• A61P7/06

  Antianaemics · CPC title

• A61P7/04

  Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents · CPC title