Claims for Patent: 8,338,376

Summary for Patent: 8,338,376

Title:	Compositions comprising variant LT-B-R-IG fusion proteins
Abstract:	This invention relates to methods of treating disease with soluble inhibitors of the lymphotoxin pathway having improved properties. This invention also relates to improved LTBR-Ig fusion proteins, and pharmaceutical compositions thereof.
Inventor(s):	Beckman; Evan (Newton, MA), Farrington; Graham K. (Acton, MA), Meier; Werner (Burlington, MA), Browning; Jeffrey L. (Brookline, MA)
Assignee:	Biogen Idec MA Inc. (Cambridge, MA)
Application Number:	12/560,257
Patent Claims:	1. A composition comprising a population of lymphotoxin-.beta. receptor (LT-.beta.-R)-Ig-fusion proteins which comprise a variant LT-.beta.-R extracellular domain of 193 or 194 amino acids in length and a variant Ig portion of 227 amino acids in length, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain, which extracellular domain is set forth in SEQ ID NO:21, and wherein the LT-.beta.-R-Ig-fusion proteins lack N-terminal pyroglutamic acid. 2. The composition of claim 1, wherein the N-terminal amino acid of the variant LT-.beta.-R-Ig fusion protein is a non-polar amino acid. 3. The composition of claim 2, wherein the non polar amino acid is either a valine, which corresponds to amino acid six of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21 or an alanine, which corresponds to amino acid five of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21. 4. The composition of claim 1, wherein the N-terminal amino acid of at least 95% of the LT-.beta.-R-Ig-fusion proteins is either a valine, which corresponds to amino acid six of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21or an alanine, which corresponds to amino acid five of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21. 5. The composition of any one of claim 1, 3 and 4, wherein the LT-.beta.-R-Ig fusion proteins are made by expressing a nucleic acid molecule comprising a nucleotide sequence encoding the extracellular domain of LT-.beta.-R set forth in SEQ ID NO:4 or SEQ ID NO:23 in a mammalian cell. 6. The composition of claim 5, wherein variant Ig portion comprises Fc regions of an IgG1 isotype. 7. The composition of claim 5, wherein the variant Ig portion comprises the amino acid sequence set forth in SEQ ID NO:2. 8. The composition of claim 5, wherein the Ig portion is non-glycosylated. 9. The composition of claim 5, wherein the LT-.beta.-R-Ig fusion proteins are made by expressing a nucleic acid molecule encoding the LT-.beta.-R-Ig fusion protein set forth in SEQ ID NO:5 in a mammalian cell. 10. The composition of claim 9, wherein the nucleic acid molecule comprises the sequence set forth in SEQ ID NO:7. 11. The composition of claim 10, wherein the step of expressing is done at manufacturing scale. 12. A composition comprising a population of lymphotoxin-.beta. receptor-immunoglobulin (LT-.beta.-R-Ig)-fusion proteins comprising a variant LT-.beta.-R extracellular domain and a variant Ig portion, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain, which extracellular domain is set forth in SEQ ID NO:21, and wherein the variant LT-.beta.-R extracellular domain is aglycosylated. 13. A composition comprising a population of lymphotoxin-.beta. receptor-immunoglobulin (LT-.beta.-R-Ig)-fusion proteins, the fusion proteins comprising a variant LT-.beta.-R extracellular domain of 193 or 194 amino acids in length and a variant Ig portion, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain, which extracellular domain is set forth in SEQ ID NO:21, and wherein the population has reduced N-terminal pyroglutamic acid formation, and reduced C-terminal heterogeneity compared to wild-type LT-.beta.-R-Ig fusion proteins. 14. The composition of claim 13, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins comprise a variant LT-.beta.-R extracellular domain as set forth the amino acid sequence of SEQ ID NO:4 or SEQ ID NO: 23. 15. The composition of claim 13, wherein the variant Ig portion comprises a mutation in the hinge region. 16. A pharmaceutical composition comprising a population of lymphotoxin-.beta. receptor (LT-.beta.-R)-Ig-fusion proteins which comprise a variant LT-.beta.-R extracellular domain of 193 or 194 amino acids in length and a variant Ig portion of 227 amino acids in length, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain, which extracellular domain is set forth in SEQ ID NO:21, and wherein the LT-.beta.-R-Ig-fusion proteins lack N-terminal pyroglutamic acid and a pharmaceutically acceptable carrier. 17. The composition of claim 16, wherein the N-terminal amino acid of the variant LT-.beta.-R-Ig fusion protein is a non-polar amino acid. 18. The composition of claim 17, wherein the non polar amino acid is either a valine, which corresponds to amino acid six of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21 or an alanine, which corresponds to amino acid five of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21. 19. The composition of claim 16, wherein the N-terminal amino acid of at least 95% of the LT-.beta.-R-Ig-fusion proteins is either a valine, which corresponds to amino acid six of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:2 or an alanine, which corresponds to amino acid five of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21. 20. The composition of claim 16, 18, or 19, wherein the LT-.beta.-R-Ig fusion proteins are made by expressing a nucleic acid molecule comprising a nucleotide sequence encoding the extracellular domain of LT-.beta.-R set forth in SEQ ID NO:4 in a mammalian cell. 21. A method of treating an autoimmune disorder comprising administering the pharmaceutical composition of claim 20 to a subject in need thereof, wherein the autoimmune disorder is rheumatoid arthritis. 22. A pharmaceutical composition comprising a population of lymphotoxin-.beta. receptor-immunoglobulin (LT-.beta.-R-Ig)-fusion proteins, the fusion proteins comprising a variant LT-.beta.-R extracellular domain of 193 or 194 amino acids in length and a variant Ig portion, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain, which extracellular domain is set forth in SEQ ID NO:21, and wherein the population has reduced N-terminal pyroglutamic acid formation and reduced C-terminal heterogeneity compared to wild-type LT-.beta.-R-Ig fusion proteins and a pharmaceutically acceptable carrier. 23. The pharmaceutical composition of claim 22, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins comprise a variant LT-.beta.-R extracellular domain as set forth the amino acid sequence of SEQ ID NO:4 or SEQ ID NO: 23. 24. The pharmaceutical composition of claim 22, wherein the variant Ig portion comprises a mutation in the hinge region. 25. A pharmaceutical composition, comprising a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:5. 26. A method of treating an autoimmune disorder comprising administering the pharmaceutical composition of claim 25 to a subject in need thereof, wherein the autoimmune disorder is rheumatoid arthritis. 27. The method of claim 26, wherein the pharmaceutical composition is administered to the subject at a dose of from about 0.6 to 3 mg/kg biweekly. 28. The method of claim 26, wherein the pharmaceutical composition is administered subcutaneously. 29. An isolated polypeptide comprising a variant LT-.beta.-R extracellular domain of 193 or 194 amino acids in length and a variant Ig portion of 227 amino acids in length, wherein the polypeptide is missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain, which extracellular domain is set forth in SEQ ID NO:21 and wherein the polypeptide lacks N-terminal pyroglutamic acid. 30. The isolated polypeptide of claim 29, wherein the N-terminal amino acid is a non-polar amino acid. 31. The isolated polypeptide of claim 30, wherein the non polar amino acid is either a valine, which corresponds to amino acid six of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21 or an alanine, which corresponds to amino acid five of the mature form of the wild type LT-.beta.-R extracellular domain of SEQ ID NO:21. 32. The isolated polypeptide of claim 29, which is made by expressing a nucleic acid molecule comprising a nucleotide sequence encoding the extracellular domain of LT-.beta.-R set forth in SEQ ID NO:23 SEQ ID NO:4 or in a mammalian cell. 33. An isolated nucleic acid molecule encoding the polypeptide of any one of claims 29-32. 34. The isolated nucleic acid molecule of claim 33, which nucleic acid molecule comprises of the nucleotide sequence set forth in SEQ ID NO:7. 35. A vector comprising the nucleic acid molecule of claim 34. 36. An isolated or cultured host cell expressing the vector of claim 35. 37. The cell of claim 36, which is a Chinese Hamster Ovary (CHO) cell. 38. A process for making a composition comprising a population of lymphotoxin-.beta. receptor (LT-.beta.-R)-Ig-fusion proteins which comprise a variant LT-.beta.-R extracellular domain and a variant Ig portion, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain set forth in SEQ ID NO:21, the process comprising, expressing a nucleic acid molecule encoding the LT-.beta.-R-Ig fusion protein set forth in SEQ ID NO:8 in a mammalian cell, obtaining the population from the culture supernatant, and, optionally, purifying the supernatant, to thereby obtain a composition comprising a population of lymphotoxin-.beta. receptor (LT-.beta.-R)-Ig-fusion proteins which comprise a variant LT-.beta.-R extracellular domain and a variant Ig portion, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R portion set forth in SEQ ID NO:21. 39. The process of claim 38, wherein the nucleic acid molecule comprises the nucleotide sequence set forth in SEQ ID NO:7. 40. The process of claim 38, wherein the nucleic acid molecule consists of the nucleotide sequence set forth in SEQ ID NO:7. 41. A method of treating rheumatoid arthritis in a human subject, in need thereof, the method comprising administering to the subject a dose of LT-.beta.-R-Ig fusion protein, wherein the dose is sufficient to maintain an average concentration of from about 0.14 ug/ml to about 3.5 ug/ml in the serum of the subject. 42. A method of treating rheumatoid arthritis in a human subject, in need thereof, the method comprising administering to the subject a dose of LT-.beta.-R-Ig fusion protein, wherein the dose is sufficient to maintain an average a minimal average concentration of about 0.6 ug/ml in the serum of the subject. 43. The method of claim 42, wherein the LT-.beta.-R-Ig fusion protein comprises the amino acid sequence set forth in SEQ ID NO:5. 44. The method of claim 42, wherein the concentration is achieved by administering LT-.beta.-R-Ig fusion protein at a dose of from about 0.01 to about 5 mg/kg once every 7-60 days. 45. A method of treating rheumatoid arthritis in a human subject, in need thereof, the method comprising administering to the subject a dose of LT-.beta.-R-Ig fusion protein of from about 0.6 to 3 mg/kg not more than twice every 7-30 days. 46. The method of claim 45, the method comprising administering to the subject a dose of LT-.beta.-R-Ig fusion protein of from about 0.6 to 3 mg/kg once every 7-14 days. 47. The method of claim 45, wherein administration is once every 14-30 days. 48. The method of claim 45, wherein administration is once every 28-60 days. 49. The method of claim 45, wherein administration is once every 7-30 days. 50. A method of treating an autoimmune disorder in a human subject, the method comprising administering to the subject a dose of a pharmaceutical composition comprising a population of LT-.beta.-R-Ig fusion proteins comprising a variant LT-.beta.-R extracellular domain of 193 or 194 amino acids in length, wherein at least 90% of the LT-.beta.-R-Ig-fusion proteins are missing no more than 5 amino acids from the N-terminus of the mature form of the wild type LT-.beta.-R extracellular domain set forth in SEQ ID NO:21 and wherein the dose is sufficient to maintain a minimal average concentration of about 0.6 ug/ml in the serum of the subject, wherein the autoimmune disorder is rheumatoid arthritis. 51. The method of claim 50, wherein the LT-.beta.-R-Ig fusion protein further comprises a variant Ig portion. 52. The method of claim 50, wherein the pharmaceutical composition comprises the amino acid sequence set forth in SEQ ID NO:5. 53. The method of claim 52, wherein administration is twice monthly. 54. The method of claim 52, wherein administration once monthly. 55. The method of claim 52, wherein administration is subcutaneous. 56. The method of claim 52, wherein the dose is about 1 mg/kg. 57. The method of claim 52, wherein the dose is about 3 mg/kg. 58. The method of claim 52, wherein the dose is about 1 mg/kg administered about every 7 to 20 days. 59. The method of claim 52, wherein the dose is about 3 mg/kg administered about every 14 to 30 days. 60. The method of claim 52, wherein the dose is about 1 mg/kg administered about every 14 days. 61. The method of claim 52, wherein the subject has been treated with a rheumatoid arthritis drug after being diagnosed with rheumatoid arthritis and prior to administration of the LT-.beta.-R-Ig fusion protein. 62. The method of claim 61, wherein the rheumatoid arthritis drug is chosen from the group consisting of a DMARD, an NSAID, and a corticosteroid. 63. The method of claim 61, wherein the human is a DMARD-inadequate responder. 64. The method of claim 61, wherein the rheumatoid arthritis drug is a TNF inhibitor. 65. The method of claim 61, wherein the rheumatoid arthritis drug is adalimumab (Humira.RTM.), etanercept (Enbrel.RTM.), or infliximab (Remicade.RTM.). 66. The method of claim 61, wherein LT-.beta.-R-Ig is administered in combination with the rheumatoid arthritis drug. 67. The method of claim 61, wherein the human is evaluated to determine if the response to the rheumatoid arthritis drug is inadequate prior to administration of LT-.beta.-R-Ig. 68. The method of claim 67, wherein the human is determined to have an inadequate response to the rheumatoid arthritis drug, and then the human is administered LT-.beta.-R-Ig. 69. The method of claim 61, wherein the human is asymptomatic for a first manifestation of rheumatoid arthritis and is symptomatic for a second manifestation of rheumatoid arthritis. 70. The method of claim 61, wherein LT-.beta.-R-Ig is administered in place of the rheumatoid arthritis drug. 71. The method of claim 61, wherein administration is in combination with a tumor necrosis factor (TNF) inhibitor. 72. The method of claim 71, wherein the TNF inhibitor is adalimumab (Humira.RTM.), etanercept (Enbrel.RTM.), or infliximab (Remicade.RTM.). 73. The method of claim 61, wherein the human is an anti-TNF-inadequate responder. 74. The method of claim 61, wherein administration is in combination with a non-steroidal anti-inflammatory agent (NSAID), a corticosteroid, or a disease modifying antirheumatic drug (DMARD). 75. The method of claim 61, wherein administration is in combination with methotrexate. 76. The method of claim 74, wherein the human is a DMARD-inadequate responder. 77. The isolated polypeptide of claim 29, wherein the polypeptide comprises the amino acid sequence set forth in SEQ ID NO:5.

Details for Patent 8,338,376

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Janssen Biotech, Inc.	REMICADE	infliximab	For Injection	103772	08/24/1998		2026-10-20
Immunex Corporation	ENBREL	etanercept	For Injection	103795	11/02/1998		2026-10-20
Immunex Corporation	ENBREL	etanercept	For Injection	103795	05/27/1999		2026-10-20
Immunex Corporation	ENBREL	etanercept	Injection	103795	09/27/2004		2026-10-20
Immunex Corporation	ENBREL	etanercept	Injection	103795	02/01/2007		2026-10-20