Claims for Patent: 6,858,409

Summary for Patent: 6,858,409

Title:	Nucleic acids encoding interleukin-1 inhibitors and processes for preparing interleukin-1 inhibitors
Abstract:	Compounds are disclosed having the general formula R.sub.1 --X--R.sub.2, wherein R.sub.1 and R.sub.2 are biologically active groups, at least one of which is polypeptidic. X is a non-peptidic polymeric group. R.sub.1 and R.sub.2 may be the same or different. Preferred R.sub.1 and R.sub.2 groups are interleukin-1 receptor antagonist, 30 kDa TNF inhibitor, interleukin-2 receptors and CR1 and muteins thereof. Also included are site selectively modified interleukin-1 receptor antagonist and 30 kDa TNF inhibitor.
Inventor(s):	Thompson; Robert C. (Boulder, CO), Hannum; Charles H. (Sunnyvale, CA), Eisenberg; Stephen P. (Boulder, CO), Arend; William P. (Denver, CO), Joslin; Fenneke G. (Denver, CO), Sommer; Andreas (Danville, CA)
Assignee:	Amgen Inc. (Thousand Oaks, CA) The Regents of the University of Colorado (Boulder, CO)
Application Number:	08/482,283
Patent Claims:	1. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding an interleukin-1 inhibitor (IL-1i) polypeptide, having interleukin-1 (IL-1) inhibitory activity, wherein said polypeptide is selected from the group consisting of: A) a polypeptide comprising all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P; and B) a polypeptide that is at least about 70% homologous to the amino acid sequence set forth in A). 2. A nucleic acid molecule of claim 1, wherein the nucleic acid sequence encodes a oolypeptide that is at least 80% homologous to amino acid sequence set forth in claim 1 (A). 3. A nucleic acid molecule of claim 1, wherein the nucleic acid sequence encodes a polypeptide that is at least 95% homologous to amino acid sequence set forth in claim 1 (A). 4. A nucleic acid molecule of claim 1, wherein the nucleic acid sequence encodes a polypeptide wherein (U) is M. 5. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding an interleukin-1 inhibitor (IL-1i) polypeptide, having interleukin-1 (IL-1 inhibitoty activity, wherein said polypeptide is selected from the group consisting of: A) a polypeptide comprising all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P; and B) a polypeptide that is at least about 90% homologous to the amino acid sequence set forth in A). 6. The nucleic acid molecule of claim 5, wherein the amino acid sequence of said polypeptide is at least about 90% homologous to the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R. 7. The nucleic acid molecule of claim 6, wherein (U) is nothing. 8. The nucleic acid molecule of claim 6, wherein (U) is M. 9. The nucleic acid molecule of claim 5, wherein the amino acid sequence of said polypeptide is at least about 95% homologous to the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q s P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R. 10. The nucleic acid molecule of claim 5, wherein said polypeptide comprises all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P. 11. The nucleic acid molecule of claim 10, wherein (X) is R. 12. The nucleic acid mnolecule of claim 10, wherein said polypeptide comprises the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P. 13. The nucleic acid molecule of claim 12, wherein (X) is R. 14. The nucleic acid molecule of claim 13, wherein (U) is M. 15. The nucleic acid molecule of claim 13, wherein (U) is nothing. 16. The nucleic acid molecule of claim 13, wherein said polypeptide consists of the following amino acid sequence: (U) R P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E C V M V T K F Y Q E D E wherein (U) is M or nothing. 17. The nucleic acid molecule of claim 16, wherein (U) is M. 18. The nucleic acid molecule of claim 13, wherein said nucleic acid sequence encoding said IL-1i polypeptide further comprises a sequence encoding an N-terminal secretion leader sequence. 19. The nucleic acid molecule of claim 18, wherein the leader sequence comprises all or part of the following amino acid sequence: M E I C R G L R S H L I T L L L F L F H S E T I C. 20. A recombinant host cell containing a DNA molecule comprising a nucleic acid sequence encoding an interleukin-1 inhibitor (IL-1i) polypeptide, said polypeptide being capable of inhibiting IL-1, wherein said polypeptide is selected from the group consisting of A) a polypeptide comprising all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M O A F R I W D V N O K T F Y L R N N O L V A G Y L O G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L O L E A V N I T D L S E N R K O D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D O P V S L T N M P D E G V M V T K F Y F O E D E wherein (U) is M or nothing and (X) is R or P; and B) a polypeptide that is at least about 70% homologous to the amino acid sequence set forth in A). 21. A recombinant host cell of claim 20, wherein said polypeptide is selected from the group consisting of: A) a polypeptide comprising all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S C R K S S K M Q A P R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P; and B) a polypeptide that is at least about 90% homologous to the amino acid sequence set forth in A). 22. The recombinant host cell of claim 21, wherein the amino acid sequence of said polypeptide is at least about 95% homologous to the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K H C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R. 23. The recombinant host cell of claim 21, wherein the amino acid sequence of said polypeptide is at least about 90% homologous to the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R. 24. The recombinant host cell of claim 23, wherein said host cell is not capable of glycosylation or is a non-human host cell. 25. The recombinant host cell of claim 24, wherein said host cell is selected from a yeast cell, a mouse Ltk.sup.- cell, and a Chinese hamster ovary cell. 26. The recombinant host cell of claim 24, wherein said host cell is a prokaryotic cell. 27. The recombinant host cell of claim 26, wherein said host cell is E. coil. 28. The recombinant host cell of claim 24, wherein said host cell is a eukaryotic cell. 29. The recombinant host cell of claim 28, wherein said host cell is a mammallan cell. 30. The recombinant host cell of claim 28, wherein said host cell is a Chinese hamster ovary cell. 31. The recombinant host cell of claim 23, wherein said host cell produces glycosylated IL-1i. 32. The recombinant host call of claim 23, wherein said host cell produces nonglycosylated IL-1i. 33. The recombinant host cell of claim 23, wherein (U) is nothing. 34. The recombinant host cell of claim 23, wherein (U) is M. 35. The recombinant host cell of claim 21, wherein said polypeptide comprises all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R X Q D K H F A F I R S D E G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E Wherein (U) is M or nothing and (X) is R or P. 36. The recombinant host call of claim 35, wherein (X) is R. 37. The recombinant host cell of claim 35, wherein said polypeptide comprises the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P. 38. The recombinant host cell of claim 37, wherein (X) is R. 39. The recombinant host cell of claim 38, wherein (U) is M. 40. The recombinant host cell of claim 38, wherein (U) is nothing. 41. The recombinant host cell of claim 38, wherein said host cell is not capable of glycosylation or is a non-human host cell. 42. The recombinant host cell of claim 41, wherein said host cell is selected from a yeast cell, a mouse Ltk.sup.- cell, and a Chinese hamster ovary cell. 43. The recombinant host cell of claim 41, wherein said host cell is a prokaryotic cell. 44. The recombinant host call of claim 43, wherein said host cell is E. coli. 45. The recombinant host cell of claim 41, wherein said host cell is a eukaryotic cell. 46. The recombinant host cell of claim 45, wherein said host cell is a mammalian cell. 47. The recombinant host cell of claim 45, wherein said host cell is a Chinese hamster ovary cell. 48. The recombinant host cell of claim 38, wherein said host cell produces glycosylated IL-1i. 49. The recombinant host cell of claim 38, wherein said host cell produces nonglycosylated IL-1 i. 50. The recombinant host cell of claim 38, wherein said DNA molecule comprises a heterologous promoter operatively linked to said nucleic acid sequence encoding said IL-1i polypeptide. 51. The recombinant host cell of claim 38, wherein said nucleic acid sequence encoding said IL-1i polypeptide further comprises a sequence encoding an N-terminal secretion leader sequence. 52. The recombinant host cell of claim 51, wherein the leader sequence comprises all or part of the following amino acid sequence: M E I C R G L R S H L I T L L L F L F H S E T I C. 53. The recombinant host cell of claim 38, wherein said polypeptide consists of the following amino acid sequence: (U) R P S G R K S S K M Q A E R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing. 54. The recombinant host cell of claim 53, wherein (U) is M. 55. The recombinant host cell of claim 54, wherein said host call produces nonglycosylated IL-1i. 56. The recombinant host cell of claim 55, wherein said host cell is E. coli. 57. The recombinant host cell of claim 53, wherein said host cell is a Chinese hamster ovary host cell. 58. The recombinant host cell of claim 53, wherein said host cell is a eukaryotic host cell. 59. The recombinant host cell of claim 53, wherein said host cell produces glycosylated IL-1i. 60. The recombinant host cell of claim 53, wherein said host cell is a mammalian host cell. 61. The recombinant host cell of claim 53, wherein said host cell is a prokaryotic host cell. 62. A process for preparing an interleukin-1 inhibitor (IL-1i) polypeptide, comprising producing the recombinant IL-1i polypeptide in a host cell according to claim 20 under suitable conditions to express the recombinant DNA molecule contained therein to produce the recombinant polypeptide. 63. The process of claim 62, further comprising harvesting the IL-1i polypeptide. 64. A process of claim 63, wherein said recombinant polypeptide is selected from the group consisting of: A) a potypeptide comprising all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P; and B) a polypeptide that is at least about 90% homologous to the amino acid sequence set forth in A). 65. The process of claim 64, wherein the amino acid sequence of said recombinant polypeptide is at least about 95% homologous to the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P T E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R. 66. The process of claim 64, wherein the amino acid sequence of said recombinant polypeptide is at least about 90% homologous to the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I P S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R. 67. The process of claim 66, wherein said host cell is not capable of glycosylation or is a non-human host cell. 68. The process of claim 67, wherein said host cell is selected from a yeast cell, a mouse Ltk.sup.+ cell, and a Chinese hamster ovary cell. 69. The process of claim 67, wherein said host cell is a prokaryotic cell. 70. The process of claim 69, wherein said host cell is E. coli. 71. The process of claim 67, wherein said host cell is a eukaryotic cell. 72. The process of claim 71, wherein said host cell is a mammalian cell. 73. The process of claim 71, wherein said host cell is a Chinese hamster ovary cell. 74. The process of claim 66, wherein said recombinant polypeptide is glycosylated. 75. The process of claim 66, wherein said recombinant polypeptide is nonglycosylated. 76. The process of claim 66, wherein (U) is nothing. 77. The process of claim 66, wherein (U) is M. 78. The process of claim 64, wherein said recombinant polypeptide comprises all or an IL-1 inhibitory fragment of the amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S 0 S G e T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P. 79. The process of claim 78, wherein (X) is R. 80. The process of claim 79, wherein said recombinant polypeptide inhibits IL-1 induced PGE.sub.2 production. 81. The process of claim 78, wherein said recombinant polypeptide comprises the following amino acid sequence: (U) (X) P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R K Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing and (X) is R or P. 82. The process of claim 81, wherein (X) is R. 83. The process of claim 82, wherein (U) is M. 84. The process of claim 82, wherein (U) is nothing. 85. The process of claim 82, wherein said host cell is not capable of glycosylalion or is a non-human host cell. 86. The process of claim 85, wherein said host cell is selected from a yeast cell, a mouse Ltk.sup.- cell, and a Chinese hamster ovary cell. 87. The process of claim 85, wherein said host cell is a prokaryotic cell. 88. The process of claim 85, wherein said host cell is E. coli. 89. The process of claim 85, wherein said host cell is a eukaryotic cell. 90. The process of claim 89, wherein said host cell is a mammalian cell. 91. The process of claim 89, wherein said host cell is a Chinese hamster ovary cell. 92. The process of claim 82, wherein said recombinant polypeptide is glycosylated. 93. The process of claim 82, wherein said recombinant polypeptide is nonglycosylated. 94. The process of claim 82, wherein said DNA molecule comprises a heterologous promoter operatively linked to said nucleic acid sequence encoding said IL-1i polypeptide. 95. The process of claim 82, wherein said nucleic acid sequence encoding said IL-1i polypeptide further comprises a sequence encoding an N-terminal secretion leader sequence. 96. The process of claim 95, wherein the leader sequence comprises all or part of the following amino acid sequence: M E I C R G L R S H L I T L L L F L F H S E T I C. 97. The process of claim 82, wherein said recombinant polypeptide consists of the following amino acid sequence: (U) R P S G R K S S K M Q A F R I W D V N Q K T F Y L R N N Q L V A G Y L Q G P N V N L E E K I D V V P I E P H A L F L G I H G G K M C L S C V K S G D E T R L Q L E A V N I T D L S E N R X Q D K R F A F I R S D S G P T T S F E S A A C P G W F L C T A M E A D Q P V S L T N M P D E G V M V T K F Y F Q E D E wherein (U) is M or nothing. 98. The process of claim 97, wherein (U) is M. 99. The process of claim 98, wherein said recombinant polypeptide is nonglycosylated. 100. The process of claim 99, wherein said host cell is E. coli. 101. The process of claim 97, wherein said host cell is a Chinese hamster ovary host cell. 102. The process of claim 97, wherein said host cell is a eukaryotic host cell. 103. The process of claim 97, wherein said recombinant polypeptide is glycosylated. 104. The process of claim 97, wherein said host cell is a mammalian host cell. 105. The process of claim 97, wherein said host cell is a prokaryotic host cell.

Details for Patent 6,858,409

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Swedish Orphan Biovitrum Ab (publ)	KINERET	anakinra	Injection	103950	November 14, 2001	6,858,409	2022-02-22

International Patent Family for US Patent 6,858,409

Country	Patent Number	Estimated Expiration
South Africa	911981
South Africa	905593
South Africa	894052
World Intellectual Property Organization (WIPO)	9534326
World Intellectual Property Organization (WIPO)	9216221
World Intellectual Property Organization (WIPO)	8911540
United States of America	6599873