Room temperature stable lyophilized protein

Patent metadata

Field	Value
Publication number	US-11351256-B2
Application number	US-201715727457-A
Country	US
Kind code	B2
Filing date	Oct 6, 2017
Priority date	Oct 7, 2016
Publication date	Jun 7, 2022
Grant date	Jun 7, 2022

Abstract

Stable lyophilized therapeutic protein compositions and their methods of manufacture are provided. Specifically, the use of water as a solid cake plasticizer and protein stabilizer is described. Also, the inclusion of a multicomponent stabilizer comprising a larger molecular entity and a smaller molecular entity is described. Also, the inclusion of post-drying annealing under certain conditions improves protein stability. Proteins are predicted to remain stable over 24 months at 25° C.

First claim

What is claimed is:

1. A lyophilized cake comprising:
(a) a protein;
(b) a stabilizer, wherein said stabilizer comprises about 5% sucrose by weight in the pre-lyophilized formulation and arginine in a weight to weight ratio of sucrose to arginine from about 3.2:1 to about 3.4:1;
(c) a buffer; and
(d) a moisture content of from about 3% to about 6% by weight,
wherein the lyophilized cake is not collapsed.

2. The lyophilized cake of claim 1 , wherein said lyophilized cake:
(i) comprises no yellow, no brown, no black, and no red color;
(ii) is not melted and comprises no density variation; and/or
(iii) comprises a glass transition temperature above 25° C.

3. The lyophilized cake of claim 1 , comprising a moisture content of from about 3.5% to about 5.5% by weight.

4. The lyophilized cake of claim 3 , comprising a moisture content of about 4.5% by weight.

5. The lyophilized cake of claim 1 , wherein said buffer comprises histidine at about 0.34% to about 2.04% by weight.

6. The lyophilized cake of claim 1 , wherein said stabilizer is:
(a) selected from the group consisting of a polyol, a sugar, an amino acid, a salt, and combinations thereof, wherein (i) said polyol is selected from the group consisting of sorbitol, glycerol, mannitol, and a combination thereof; (ii) said sugar is selected from the group consisting of trehalose, sucrose, and a combination thereof; (iii) said amino acid is selected from the group consisting of arginine, alanine, proline, glycine, and a combination thereof; and (iv) said salt is sodium chloride; and/or
(b) present at about 19% to about 83% by weight.

7. The lyophilized cake of claim 1 , wherein said stabilizer further comprises mannitol, wherein said mannitol is present at about 8% to about 23% by weight.

8. The lyophilized cake of claim 1 , wherein said stabilizer further comprises sorbitol, wherein said sorbitol is present at about 1.3% to about 23% sorbitol by weight.

9. The lyophilized cake of claim 1 further comprising a surfactant, wherein:
(a) said surfactant is selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, polyethylene glycol (PEG), PEG 3350, and a combination thereof; or
(b) said surfactant is present at about 0.02% to about 1% by weight.

10. The lyophilized cake of claim 1 , wherein said protein is present from about 6% to about 64% by weight.

11. The lyophilized cake of claim 1 , wherein said protein is selected from the group consisting of an antigen-binding protein, an antibody, an antibody fragment, and a receptor-Fc-fusion protein.

12. The lyophilized cake of claim 1 , wherein said lyophilized cake is contained within a vial, wherein said vial comprises type I borosilicate glass and is closed with a polytetrafluorethylene coated butyl rubber stopper.

13. A lyophilized cake comprising:
(a) a protein;
(b) a stabilizer at a ratio of about 0.2 parts stabilizer to 1 part protein by weight (0.2:1) to about 1.5 parts stabilizer to 1 part protein by weight (1.5:1), wherein said stabilizer comprises arginine, sorbitol, glycerol and/or alanine; and
(c) a moisture content of from about 3% to about 6% by weight,
wherein said lyophilized cake is free of discoloration.

14. The lyophilized cake of claim 13 , wherein the ratio of the stabilizer to protein by weight is between about 0.22:1 and about 1.3:1 by weight.

15. The lyophilized cake of claim 13 , wherein said stabilizer:
(a) is selected from the group consisting of a non-reducing sugar, a sugar alcohol, an amino acid, and a combination thereof, wherein said non-reducing sugar comprises sucrose or trehalose; said sugar alcohol is selected from the group consisting of sorbitol, mannitol, and glycerol; and said amino acid is selected from the group consisting of arginine, alanine, proline, and glycine; or
(b) comprises sucrose or trehalose.

16. The lyophilized cake of claim 15 (b) further comprising arginine, mannitol or sorbitol, wherein:
(a) the ratio of sucrose to arginine is about 3.2:1 to about 3.4:1 by weight;
(b) the ratio of sucrose to mannitol is about 1.8:1 to about 2:1 by weight;
(c) the ratio of trehalose to arginine is about 3.2:1 to about 3.4:1 by weight; or
(d) the ratio of trehalose to sorbitol is about 1:1 to about 2:1 by weight.

17. The lyophilized cake of claim 13 , wherein said protein is selected from the group consisting of an antibody, an antibody fragment, and a receptor-Fc-fusion protein.

18. The lyophilized cake of claim 13 , further comprising a buffer, wherein said buffer is selected from the group consisting of citrate, phosphate, histidine, succinate, carbonate, acetate, and a combination thereof.

19. The lyophilized cake of claim 13 , wherein said lyophilized cake is contained within a vial or a syringe.

Assignees

• Regeneron Pharma

Inventors

• Tang Xiaolin
• Ludwig David Brett

Classifications

• A61K39/39591 Primary

  Stabilisation, fragmentation · CPC title

• A61K47/22

  Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones · CPC title

• A61K47/10

  Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers · CPC title

• A61K47/183

  Amino acids, e.g. glycine, EDTA or aspartame · CPC title

• A61K47/26

  Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin · CPC title