throbber
(12) United States Patent
`Heavner et a].
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`US 7,250,165 B2
`Jul. 31, 2007
`
`US007250165B2
`
`(54)
`
`(75)
`
`ANTI-TNF ANTIBODIES, COMPOSITIONS,
`METHODS AND USES
`
`Inventors: George Heavner, Malvem, PA (US);
`David M. Knight, BerWyn, PA (US);
`Jill Giles-Komar, DoWningtoWn, PA
`(US); Bernard Scallon, Collegeville,
`PA (US); David Shealy, DoWningtoWn,
`PA (US)
`
`(73)
`
`Assignee: Centocor, Inc., Malvern, PA (US)
`
`Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`USC 154(b) by 916 days.
`
`(21)
`
`(22)
`
`(65)
`
`(60)
`
`(51)
`
`(52)
`
`(58)
`
`(56)
`
`Appl. No.: 09/920,137
`
`Filed:
`
`Aug. 1, 2001
`
`Prior Publication Data
`
`US 2003/0049725 A1
`
`Mar. 13, 2003
`
`Related U.S. Application Data
`
`Provisional application No. 60/236,826, ?led on Sep.
`29, 2000, provisional application No. 60/223,360,
`?led on Aug. 7, 2000.
`
`Int. Cl.
`(2006.01)
`A61K 39/395
`(2006.01)
`C07K 16/28
`U.S. Cl. ............................. .. 424/145.1; 424/130.1;
`424/133.1; 530/388.23; 530/3873
`Field of Classi?cation Search ........... .. 530/387.3;
`424/133.1, 145.1
`See application ?le for complete search history.
`
`References Cited
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`(Continued)
`Primary Examiner4Christine J. Saoud
`Assistant Examinerilegatheesan Seharaseyon
`
`(57)
`
`ABSTRACT
`
`The present invention relates to at least one novel anti-TNF
`antibodies, including isolated nucleic acids that encode at
`least one anti-TNF antibody, TNF, vectors, host cells, trans
`genic animals or plants, and methods of making and using
`thereof, including therapeutic compositions, methods and
`devices.
`
`5 Claims, 26 Drawing Sheets
`
`Ex. 1006 - Page 1 of 98
`
`

`

`US 7,250,165 B2
`Page 2
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`Exley, A.R. et al., “Monoclonal Antibody (Mab) to Recombinant
`Human Tumour Necrosis Factor (rhTNF) in the Prophylaxis and
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`Cross, A.S. et al., “Pretreatment with Recombinant Murine Tumor
`Necrosis Factor 01 Cachectin and Murine Interleukin 1 0t Protects
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`Autocrine Growth Inhibition in a Chronic Leukemia,” J Immunol,
`143:3828-3834 (1989).
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`U937 Cells, and in Mice,” JImmunol, 143:3517-3523 (1989).
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`Necrosis Factor-. at 2.6 El Resolution,” JBiol Chem, 264:17595
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`Engelmann, Hartmut et al., “A Tumor Necrosis Factor-binding
`Protein Puri?ed to Homogeneity from Human Urine Protects Cells
`from Tumor Necrosis Factor Toxicity,” J of Bio. Chem.,
`264(20):11974-11980 (1989).
`Kawasaki, Hajime et al., “Analysis of Endotoxin Fever in Rabbits
`by Using a Monoclonal Antibody to Tumor Necrosis Factor
`(Cachectin),” Infection and Immunity, 57(10):3131-3135 (1989).
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`Factor Reduce Interleukin 15 and Interleukin 6 Appearance During
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`with a Human Monoclonal Antibody Against Lipopolysaccharide
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`

`

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`Page 3
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`
`* cited by examiner
`
`Ex. 1006 - Page 3 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 1 of 26
`
`US 7,250,165 B2
`
`NNEEIDI
`
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`omF>ZF.IATI
`
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`
`Ex. 1006 - Page 4 of 98
`
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`
`6:302
`
`Ex. 1006 - Page 4 of 98
`
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 2 0f 26
`
`US 7,250,165 B2
`
`FIG. 2A
`
`TNVS
`
`ATGGGGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTAAGA
`
`germline
`
`Q V Q L V E S G G G V
`CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTG
`
`TNVS
`
`GGTGTCCAGTGT . . . . . . . . . . . .
`
`. . . . . . . . . . . . . . . . . . . . .
`
`TNV148 (B) GGTGTCCAGTGT. . . . .A . . .
`
`. . .
`
`. . . . . . . . . . . . . . . . . . . . .
`
`V Q P G R S L R L S C A A S G
`I germline GTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGA
`
`TNVS
`
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`F S S Y A M H W V R Q A P
`F 'I‘
`germline TTCACCTTCAGTAGC'I‘A'I‘GCTA'I‘GCACTGGG'I‘CCGCCAGGCTCCA
`
`TNVléL, 15 . . . . . . . . . . . . . . . . .
`
`. . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNV148 (B) . . . .T . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`'I‘NV196
`
`. . . . . . . . . . . . . . . . . . . .C . . . . . . . . . . . . . . . . . . . . . . . .
`
`G K G L E W V A V I S Y D G S
`germline GGCAAGGGGCTGGAGTGGG‘I‘GGCAGTTATATCA'I‘ATGATGGAAGC
`
`TNV14
`
`. . . . . . . . . . . . . . . . . . . . . . . .A. . . .C.T . . . . . . . . . . . .T
`
`TNVl5
`
`. . . . . . . . . . . . . . . . . . . . . . . .T . . . . . .T . . . . . . . . . . . .‘I‘
`
`TNV148 (B) . . . . .C . . . . . . . . . . . . . . . . . .T. . . .G . . . . . . . . . . . . . . .
`
`TNV196
`
`. . . . . . . . . . . . . . . . . . . . . . . .‘I‘ . . . . . . . . . . . . .
`
`. . . . . . .
`
`N K Y Y A D S V K G R F T I S
`germline AATAAATACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCC
`
`TNV14
`
`.GC. . .A.G. . . . .G . . . . . . . . . . . . .A . . . . . . . . . . . . . . . .
`
`TNVl5
`
`. .C. . .A.G . . . . . . . . . . . . . . . . . . . . . . .C . . . . . . . . . . . .
`
`TNV148 (B) . . .
`
`. . .A.G . . . . . . . . . . . . . . . . . . . . . .
`
`. . .
`
`. . . . . . . . . . .
`
`TNV196
`
`. . .
`
`. . .A.G.C . . . . . . . . . . . . . . . . . . . . . . . . . . . .G. . . . .
`
`Ex. 1006 - Page 5 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 3 0f 26
`
`US 7,250,165 B2
`
`FIG. 2B
`
`R D N S K N T L Y L Q M N S L
`germline AGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTG
`
`TNV14
`
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNV15
`
`.
`
`. . . . . . . . . . . . . . . . .G . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNT/148
`
`.
`
`.
`
`. . . . . . .C . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`. . . . . . . . .
`
`TNVl48B
`
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNV196
`
`. . . . . . . . . . . . . . . . . .
`
`. . . . . . .T . . . . . . . . . . . . . . . . . . .
`
`R A E D T A V Y Y C A R
`germline AGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGA
`
`TNVl4, l5
`
`. . . . . . . . . . . . . . . . . . . . . . .
`
`. . . . . . . . . . . . .GATCGAGGT
`
`TNV148 (B) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .A
`
`TNVl96
`
`. . . . . . . . . . . . . . . . . . . . . .T . . . . . . . . . . . . . . . . . . . . . .
`
`germline
`TNV14
`
`Y Y Y Y Y G M D V W
`TACTACTACTACTACGGTATGGACGTCTGG
`
`ATATCAGCAGGTGGAA . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`.
`
`TNV15
`
`G.C .
`
`. . . . . . . . . . .A.T. .T . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNV148 (B) . . .G . . . . . . . . . . .A . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNV196
`
`. .TGG . . . . . . . . . .A . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`G Q G T T V T V S S
`germline GGGCAAGGGACCACGGTCACCGTCTCCTCAG
`
`TNV14
`
`. .C . . . . . .
`
`.
`
`. . . . . . . . . . . . . . . . . . . . .
`
`TNVl5
`
`. .C . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`TNV148 (B) . .C . . . . . . . . . . . . . . . . . . . . . . .
`
`. . . .
`
`.
`
`TNV196
`
`. .C. .G . . . .
`
`. . . . . . . . . . . . . . . .
`
`.
`
`. . . .
`
`Ex. 1006 - Page 6 of 98
`
`

`

`U.S. Patent
`
`Jul. 31,2007
`
`Sheet 4 0f 26
`
`US 7,250,165 B2
`
`FIG. 3
`
`TNVs
`
`ATGGAAGCCCCAGCTCAGCTTCTCTTCCTCCTGCTACTCTGGCTC
`
`germline
`TNVS
`
`E I V L T Q S P A T
`GAAATTGTGTTGACACAGTCTCCAGCCACC
`
`CCAGATACCACCGGA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`germline
`TNVs
`
`L S L S P G E R A T L S C R A
`CTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCC
`
`u u n Q u q ~ u a ~ u o s u u ~ a - Q o ¢ o ¢ v u u u - 1 I c o n u n u u a I n o . n 0 a
`
`germline
`TNV14, l5
`TNV148,196
`
`S Q S V S S Y L A W Y Q Q K P
`AGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCT
`
`u u o a u o a n a n a a n c u a a a o a u . u - o a n o n u u Q u Q p o u a n a o a o a o
`
`.ov
`
`. . . . . .
`
`germline
`TNVs
`
`G Q A P R L L I Y D A S N R A
`GGCCAGGCTCCCAGGCTCCTCATCTATGATGCATCCAACAGGGCC
`
`~ v ¢ ~ u v a . u . o v o » a u v Q o a o 0 o u u - . . u Q ~ c - Q a I o u . - o a - ¢ 0
`
`germline
`TNVs
`
`'1'_ G I P A R F s G s G s G T D
`ACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGAC
`
`y n n o n n a n o a o o - a - Q . u u ¢ . . . a v v o u u o e n u u o a q n a u ‘ a ' a C
`
`germline
`TNVS
`
`F T L T I S S L E P E D F A V
`TTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTT
`
`v - ¢ a I u u u - - a n o - a - u . n
`
`n n . - u . u u o u
`
`v Q ‘ ~ u ¢ - 1 c c v u v a o n
`
`Germline
`TNVS
`
`Y Y c Q Q R g N W P P F '1' F G
`TATTACTGTCAGCAGCGTAGCAACTGGCCTCCATTCACTTTCGGC
`
`. - a u - - Q c o - 0 - . o u - . v ‘ ~ u - - I u - c o u Q u o p - n a . a a o u Q u - -
`
`gerline
`TNVs
`
`P G T K V D I K R
`CCTGGGACCAAAGTGGATATCAAACGT
`
`a o u . u a u Q - Q n a - - - . u u n a v ~ . o u 1
`
`Ex. 1006 - Page 7 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 5 0f 26
`
`US 7,250,165 B2
`
`FIG. 4
`
`MGFGLSWVFLVALLRGVQC
`
`signal
`
`FRl
`
`CDRl
`
`PR2
`
`CDRZ
`
`u o a a o a a a o u . n a o v u a a o o a
`
`n o a n o u o 0 0
`
`germline
`TNVS
`
`germline
`TNVS
`TNV148 (B)
`
`germline
`TNVS
`
`SYAMH
`
`germline
`TNVs
`TNV148 (B)
`
`WVRQAPGKGLEWVA
`
`. . . . . . .N. . . . . .
`
`germline
`TNVl4
`TNV15
`TNV148 (B)
`TNV196
`
`VISYDGSNKYYADSVKG
`
`I .L. . . .S.K . . . . . .D
`
`F.L . . . . . .K . . . . . . .
`
`FM . . . . . . .K . . . .
`
`. . .
`
`F . . . . . . . .KS . . .
`
`.
`
`. .
`
`germline
`TNV14
`TNV15
`TNV148
`TNVl48B
`TNV196
`
`germline
`TNV14
`TNV15
`TNVl48 (B)
`TNV196
`
`RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
`
`FR3
`
`. . . . . . . . . . .A. .
`
`. . . . . . . . . . . . . .
`
`. . . .
`
`. . . . . . . . P. . . . .
`
`. . . . . . .
`
`. . . . . . . . . . .
`
`V . . . . . . . . .F . . . . . . . . . . . . .F
`
`— — — — — - ——YYYYYGMDV
`
`DRGISAGGN . . . . . .
`
`. .
`
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`
`. . .
`
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`
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`
`.G.
`
`N . . . . . . . .
`
`CDR3
`
`germline
`TNVS
`
`WGQGTTVTVSS
`
`¢ u u u a u ¢ a a 0 a
`
`Ex. 1006 - Page 8 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 6 0f26
`
`US 7,250,165 B2
`
`FIG. 5
`
`TNVS
`
`MEAPAQLLFLLLLWLPDTTG
`
`signal
`
`EIVLTQSPATLSLSPGERATLSC
`
`FRl
`
`CDRl
`
`FR2
`
`CDRZ
`
`PR3
`
`CDR3
`
`germline
`TNVS
`
`germline
`TNV14
`TNV15
`TNV148 (B)
`TNV196
`
`germline
`TNVS
`
`germline
`TNVS
`
`germline
`TNVS
`
`germline
`TNVS
`
`germline
`TNVS
`
`Ex. 1006 - Page 9 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 7 0f 26
`
`US 7,250,165 B2
`
`0 Enhancer
`
`Ex. 1006 - Page 10 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 8 0f 26
`
`US 7,250,165 B2
`
`mm
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`Ex. 1006 - Page 11 of 98
`
`

`

`U.S. Patent
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`Ex. 1006 - Page 12 of 98
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`Ex. 1006 - Page 12 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 10 0f 26
`
`US 7,250,165 B2
`
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`Ex. 1006 - Page 13 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 11 0f 26
`
`US 7,250,165 B2
`
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`Ex. 1006 - Page 14 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 12 0f 26
`
`US 7,250,165 B2
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`Ex. 1006 - Page 15 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
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`Ex. 1006 - Page 16 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 14 0f 26
`
`US 7,250,165 B2
`
`FIG. 9A
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`Ex. 1006 - Page 17 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 15 of 26
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`US 7,250,165 B2
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`Ex. 1006 - Page 18 of 98
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`Ex. 1006 - Page 18 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 16 6f 26
`
`US 7,250,165 B2
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`Ex. 1006 - Page 19 of 98
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`

`

`U.S. Patent
`
`Jul. 31, 2007
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`Sheet 17 of 26
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`US 7,250,165 B2
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`EX. 1006 - Page 20 of 98
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`Ex. 1006 - Page 20 of 98
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 18 of 26
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`US 7,250,165 B2
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`EX. 1006 - Page 21 of 98
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`Ex. 1006 - Page 21 of 98
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`
`
`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 19 of 26
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`US 7,250,165 B2
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`Ex. 1006 - Page 22 of 98
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`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 20 of 26
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`US 7,250,165 B2
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`Ex. 1006 - Page 23 of 98
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`Ex. 1006 - Page 23 of 98
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`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 21 0f 26
`
`US 7,250,165 B2
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`Ex. 1006 - Page 24 of 98
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`Ex. 1006 - Page 24 of 98
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`

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`U.S. Patent
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`Ex. 1006 - Page 25 of 98
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`Ex. 1006 - Page 26 of 98
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`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 24 0f 26
`
`US 7,250,165 B2
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`Ex. 1006 - Page 27 of 98
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`Ex. 1006 - Page 27 of 98
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`Ex. 1006 - Page 28 of 98
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`
`

`

`U.S. Patent
`
`Jul. 31, 2007
`
`Sheet 26 of 26
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`US 7,250,165 B2
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`Ex. 1006 - Page 29 of 98
`
`

`

`US 7,250,165 B2
`
`1
`ANTI-TNF ANTIBODIES, COMPOSITIONS,
`METHODS AND USES
`
`This application is based in part on, and claims priority to,
`US. Provisional 60/223,360 filed Aug. 7, 2000 and 60/236,
`826 filed Sep. 29, 2000, each of which is entirely incorpo-
`rated herein by reference.
`
`BACKGROUND OF THE INVENTION
`
`1. Field of the Invention
`
`including
`invention relates to antibodies,
`The present
`specified portions or variants, specific for at least one tumor
`necrosis factor alpha (TNF) protein or fragment thereof, as
`well as nucleic acids encoding such anti-TNF antibodies,
`complementary nucleic acids, vectors, host cells, and meth-
`ods of making and using thereof,
`including therapeutic
`formulations, administration and devices.
`2. Related Art
`
`5
`
`10
`
`15
`
`2
`
`Surg. 762670-671 (1989); Debets et al., Second Vienna
`Shock Forum, p. 463-466 (1989); Simpson et al., Crit. Care
`Clin. 5227-47 (1989)), including fever, malaise, anorexia,
`and cachexia. Endotoxin strongly activates monocyte/mac-
`rophage production and secretion of TNF alpha and other
`cytokines (Kombluth et al., J. Immunol. 13722585-2591
`(1986)). TNF alpha and other monocyte-derived cytokines
`mediate the metabolic and neurohormonal responses to
`endotoxin (Michie et al., New Engl. J. Med. 31821481-1486
`(1988)). Endotoxin administration to human volunteers pro-
`duces acute illness with flu-like symptoms including fever,
`tachycardia,
`increased metabolic rate and stress hormone
`release (Revhaug et al., Arch. Surg. 1232162-170 (1988)).
`Circulating TNF alpha increases in patients suffering from
`Gram-negative sepsis (Waage et al., Lancet 12355-357
`(1987); Hammerle et al., Second Vienna Shock Forum, p.
`715-718 (1989); Debets et al., Crit. Care Med. 172489-497
`(1989); Calandra et al., J. Infect. Dis. 1612982-987 (1990)).
`Thus, TNF alpha has been implicated in inflammatory
`diseases, autoimmune diseases, viral, bacterial and parasitic
`infections, malignancies, and/or neurodegenerative diseases
`and is a useful
`target for specific biological
`therapy in
`diseases, such as rheumatoid arthritis and Crohn’s disease.
`Beneficial effects in open-label trials with a chimeric mono-
`clonal antibody to TNF alpha (cA2) have been reported with
`suppression of inflammation and with successful retreatment
`after relapse in rheumatoid arthritis (Elliott et al., Arthritis
`Rheum. 3621681-1690 (1993); and Elliott et al., Lancet
`34421125-1127 (1994)) and in Crohn’s disease (Van Dulle-
`men et al., Gastroenterology 1092129-135 (1995)). Benefi-
`cial results in a randomized, double-blind, placebo-con-
`trolled trial with cA2 have also been reported in rheumatoid
`arthritis with suppression of inflammation (Elliott et al.,
`Lancet 34421105-1110 (1994)).
`Antibodies to a “modulator” material which was charac-
`
`terized as cachectin (later found to be identical to TNF) were
`disclosed by Cerami et al. (EPO Patent Publication 0212489,
`Mar. 4, 1987). Such antibodies were said to be useful in
`diagnostic immunoassays and in therapy of shock in bacte-
`rial
`infections. Rubin et al.
`(EPO Patent Publication
`0218868, Apr. 22, 1987) disclosed monoclonal antibodies to
`human TNF,
`the hybridomas secreting such antibodies,
`methods of producing such antibodies, and the use of such
`antibodies in immunoassay of TNF. Yone et al. (EPO Patent
`Publication 0288088, Oct. 26, 1988) disclosed anti-TNF
`antibodies, including mAbs, and their utility in immunoas-
`say diagnosis of pathologies,
`in particular Kawasaki’s
`pathology and bacterial infection. The body fluids of patients
`with Kawasaki’s pathology (infantile acute febrile mucocu-
`taneous lymph node syndrome; Kawasaki, T., Allergy
`162178 (1967); Kawasaki, T., Shonica (Pediatrics) 262935
`(1985)) were said to contain elevated TNF levels which were
`related to progress of the pathology (Yone et al., supra).
`Other investigators have described mAbs specific for
`recombinant human TNF which had neutralizing activity in
`vitro (Liang, C-M. et al. (Biochem. Biophys. Res. Comm.
`1372847-854 (1986); Meager, A. et al., Hybridoma 62305-
`311 (1987); Fendly et al., Hybridoma 62359-369 (1987);
`Bringman, T. S. et al., Hybridoma 62489-507 (1987); Hirai,
`M. et al., J. Immunol. Meth. 96:57-62 (1987); Moller, A. et
`al. (Cytokine 22162-169 (1990)). Some of these mAbs were
`used to map epitopes of human TNF and develop enzyme
`immunoassays (Fendly et al., supra; Hirai et al., supra;
`Moller et al., supra) and to assist in the purification of
`recombinant TNF (Bringman et al., supra). However, these
`studies do not provide a basis for producing TNF neutral-
`izing antibodies that can be used for in vivo diagnostic or
`
`Ex. 1006 - Page 30 of 98
`
`20
`
`25
`
`30
`
`35
`
`40
`
`TNF alpha is a soluble homotrimer of 17 kD protein
`subunits
`(Smith et al.,
`J. Biol. Chem. 26226951-6954
`(1987)). A membrane-bound 26 kD precursor form of TNF
`also exists (Kriegler et al., Cell 53:45-53 (1988)). For
`reviews of TNF, see Beutler et al., Nature 3202584 (1986);
`Old, Science 2302630 (1986); and Le et al., Lab. Invest.
`562234 (1987).
`Cells other than monocytes or macrophages also produce
`TNF alpha. For example, human non-monocytic tumor cell
`lines produce TNF alpha (Rubin et al., J. Exp. Med. 164:
`1350 (1986); Spriggs et al., Proc. Natl. Acad. Sci. USA
`8426563 (1987)). CD4+ and CD8+ pe

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