Claims for Patent: 5,643,566

Summary for Patent: 5,643,566

Title:	Formulation processes for lipophilic proteins
Abstract:	Highly stable, pharmaceutical compositions comprising a therapeutically effective amount of a biologically active recombinant lipophilic protein such as human .beta.-interferon and interleukin-2 dissolved in a non-toxic, inert, therapeutically compatible aqueous-based carrier at pH ranges of from about 6.8 to 7.8 and about 8.5 to 10, which formulations contain a stabilizer for the protein selected from the group comprising human serum album, human serum albumin and dextrose or human plasma protein fraction, and at a pH range of from about 2 to 4, which latter formulations optionally contain a carbohydrate and/or protein stabilizer, are disclosed. Further, preferred formulation processes to prepare said compositions are described which avoid highly alkaline conditions.
Inventor(s):	Hanisch; Wolfgang H. (Balmoral Heights, AU), Fernandes; Pete M. (Walnut Creek, CA), Taforo; Terrance (Oakland, CA), Thomson; James W. (Albany, CA)
Assignee:	Cetus Corporation (Emeryville, CA)
Application Number:	08/474,769
Patent Claims:	1. A process for formulating a lipophilic protein selected from an unglycosylated recombinant human interferon-.beta. (IFN-.beta.) and an unglycosylated recombinant human interleukin-2 (IL-2) that has been recovered from a bacterial host transformed to produce it wherein the cell wall and cell membrane of the host was disrupted and the protein in the disruptate was isolated and purified up to the point of final diafiltration or desalting, comprising the steps of (a) diafiltering or desalting said lipophilic protein at a pH range of about 8.5 to 10.0 employing a low ionic strength elution buffer containing a transfer component; (b) lowering the pH of the diafiltrate or desalted pool with an appropriate acid agent to a pH from about 2 to about 4; and (c) removing the precipitated transfer component by centrifugation and/or filtration. 2. A process according to claim 1 further comprising the steps of: (d) adding to the protein pool a carbohydrate stabilizer; and (e) lyophilizing the resulting composition at the pH range of about 2 to about 4. 3. A process according to claim 1 further comprising the steps of: (d) adding to the protein pool a stabilizer for the protein which has previously been adjusted to a pH of about 2 to about 4; (e) incubating the mixture at said pH range of about 2 to about 4 from about five to forty-five minutes; and (f) raising the pH of the protein solution to 6.8 to 7.8 with an appropriate basic agent. 4. A process according to claim 3 further comprising the steps of: (g) lyophilizing the protein composition at the pH range of 6.8 to 7.8; and (h) optionally reconstituting said lyophilized composition. 5. A process according to claim 4 wherein the protein stabilizer is HSA or PPF, the pH range for step (a) is 9 to 9.5; the pH range for steps (b) and (d) is 3 to 4; and the pH range for steps (f) and (g) is 7.2 to 7.6. 6. A process according to claim 5 wherein the pH range for step (a) is 9 to 9.2; the pH for steps (b) and (d) is about 3.5 and the pH for steps (f) and (g) is about 7.5. 7. A process according to claim 3 further comprising the steps of: (g) adding a polyol to the protein solution; (h) pre-filtering and sterile filtering the solution; (i) lyophilizing the solution; and (j) optionally reconstituting said lyophilized composition. 8. A process according to claim 7 wherein the protein stabilizer is HSA or PPF, and wherein the polyol is mannitol or dextrose. 9. A process according to claim 3 wherein the stabilizer is human serum albumin or human plasma protein fraction, and the protein is IL-2 or .beta.-HIFN. 10. A process according to claim 4 wherein the protein is IL-2 and the stabilizer is human serum albumin. 11. A process according to claim 4 wherein the protein is .beta.-HIFN and the stabilizer is human plasma protein fraction. 12. A process according to claim 11 wherein the .beta.-HIFN is IFN-.beta..sub.ser17. 13. A process according to claim 4 wherein the transfer component of step (a) is at a concentration range of 0.5 to about 2% and is selected from the group co, rising a fatty acid salt having 10 to 13 carbon atoms, urea at a concentration of 5-7M, or guanidine hydrochloride at a concentration of 5-7M. 14. A process according to claim 13 wherein the transfer component is at a concentration range of about 0.1% to about 1% and is a laurate salt. 15. A process according to claim 14 wherein said laurate salt is sodium laurate. 16. A process according to claim 4 wherein the mixture is incubated at pH 3 to 4 for about five to 15 minutes. 17. A process of claim 2 wherein the carbohydrate stabilizer is mannitol or dextrose. 18. A process of claim 1, wherein the recombinant human IFN-.beta. is IFN-.beta..sub.ser17. 19. A process for formulating a lipophilic protein selected from an unglycosylated recombinant human interferon-.beta. (IFN-.beta.) and an unglycosylated recombinant human interleukin-2 (IL-2) that has been recovered from a bacterial host transformed to produce it wherein the cell wall and cell membrane of the host was disrupted and the protein in the disruptate was isolated and purified, comprising the steps of: (a) adjusting the pH of the medium in which the protein is contained to about 2 to 4; (b) adding to the protein medium a stabilizer for the protein which has been previously adjusted to a pH of about 2 to 4; and (c) lyophilizing the resulting composition at about pH 2 to 4, wherein the recombinant human IFN-.beta. is IFN-.beta..sub.ser17. 20. A process for formulating a lipophilic protein selected from an unglycosylated recombinant human interferon-.beta. (IFN-.beta.) and an unglycosylated recombinant human interleukin-2 (IL-2), isolated from a bacterial host transformed to produce it and purified comprising the steps of (a) adjusting the pH of the medium in which the lipophilic protein is contained to about 2 to 4; (b) adding to the protein medium a protein stabilizer for the lipophilic protein which has been previously adjusted to a pH of about 2 to 4; and (c) raising the pH of the resulting composition to 6.8 to 7.8, wherein the recombinant human IFN-.beta. is IFN-.beta..sub.ser17. 21. A process for formulating a lipophilic protein selected from an unglycosylated recombinant human interferon-.beta. (IFN-.beta.) and an unglycosylated recombinant human interleukin-2 (IL-2) which has been extracted and isolated from a bacterial host transformed to produce it and purified, comprising the steps of (a) combining the protein with a protein stabilizer and adjusting the pH of the combination to about 2 to 4; and (b) raising the pH of the resulting composition to 6.8 to 7.8, wherein the recombinant human IFN-.beta. is IFN-.beta..sub.ser17.

Details for Patent 5,643,566

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Grifols Therapeutics Llc	ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5	albumin (human)	For Injection	101138	10/21/1942		2014-07-01
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		2014-07-01
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		2014-07-01
Grifols Biologicals Llc	ALBUTEIN	albumin (human)	Injection	102478	08/15/1978		2014-07-01
Instituto Grifols, S.a.	HUMAN ALBUMIN GRIFOLS	albumin (human)	Injection	103352	02/17/1995		2014-07-01
Instituto Grifols, S.a.	HUMAN ALBUMIN GRIFOLS	albumin (human)	Injection	103352	06/11/2003		2014-07-01