(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(19) World Intellectual Property
`Organization
`International Bureau
`
`(43) International Publication Date
`
`26 February 2004 (26.02.2004)
`
`
`
`(10) International Publication Number
`
`WO 2004/016286 A2
`
`(51) International Patent Classification7:
`
`A61K 39/395
`
`(21) International Application Number:
`PCT/IB2003/004502
`
`(22) International Filing Date: 15 August 2003 (15.08.2003)
`
`(25) Filing Language:
`
`English
`
`(81) Designated States (national): AE, AG, AL, AM, AT, AU,
`AZ, BA, BB, BG, BR, BY, BZ, CA, CH, CN, CO, CR, CU,
`CZ, DE, DK, DM, DZ, EC, EE, ES, FI, GB, GD, GE, GH,
`GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC,
`LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW,
`MX, MZ, NO, NZ, OM, PH, PL, PT, RO, RU, SC, SD, SE,
`SG, SK, SL, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ,
`VC, VN, YU, ZA, ZM, ZW.
`
`(26) Publication Language:
`
`English
`
`(34)
`
`(30) Priority Data:
`10/222,140
`
`16 August 2002 (16.08.2002)
`
`US
`
`(71) Applicant (for all designated States except US): ABBOTT
`LABORATORIES CBERMUDA) LTD. [—/—]; Claren-
`don House, 2 Church Street, Hamilton HM 11 (BM).
`
`(72) Inventors; and
`KRAUSE,
`only):
`(for US
`(75) Inventors/Applicants
`Hans-Juergen [DE/DE]; Dieselstrasse 33, 67551 Worms
`(DE). BAUST, Lisa [DE/DE]; Stamitzstrasse 8, 68167
`Mannheim (DE). DICKES, Michael [DE/DE]; Schafer—
`gasse 58, 67127 Rodersheim—Gronau (DE).
`
`
`
`2004/016286A2||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||l
`
`Designated States (regional): ARIPO patent (GH, GM,
`KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZM, ZW),
`Eurasian patent (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM),
`European patent (AT, BE, BG, CH, CY, CZ, DE, DK, EE,
`ES, FI, FR, GB, GR, HU, IE, IT, LU, MC, NL, PT, RO,
`SE, SI, SK, TR), OAPI patent (BF, BJ, CF, CG, CI, CM,
`GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG).
`
`Published:
`
`without international search report and to be republished
`upon receipt of that report
`
`For two—letter codes and other abbreviations, refer to the ”Guid—
`ance Notes on Codes and Abbreviations " appearing at the begin-
`ning of each regular issue of the PCT Gazette.
`
`AMGEN INC.
`
`Exhibit 1047
`
`O (54) Title: FORMULATION OF HUMAN ANTIBODIES FOR TREATING TNF—ALPHA ASSOCIATED DISORDERS
`W (57) Abstract: FORMULATION OF HUMAN ANTIBODIES FOR TREATING TNF—ALPHA ASSOCIATED DISORDERS
`
`Ex. 1047 — Page 1 of 46
`
`Ex. 1047 - Page 1 of 46
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`AMGEN INC.
`Exhibit 1047
`
`

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`WO 2004/016286
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`PCT/IB2003/004502
`
`-1-
`
`FORMULATION OF HUMAN ANTIBODIES FOR TREATING TNF-on
`ASSOCIATED DISORDERS
`
`Background of the Invention
`
`Tumor necrosis factor oc (TNFOL) is a cytokine produced by numerous cell types,
`
`including monocytes and macrophages, that was originally identified based on its
`capacity to induce the necrosis of certain mouse tumors (see e.g., Old, L. (1985) Science
`23_0:630-632). Subsequently, a factor termed cachectin, associated with cachexia, was
`shown to be the same molecule as TNFOL. TNFOL has been implicated in mediating
`
`10
`
`shock (see e.g., Beutler, B. and Cerami, A. (1988) Annu. Rev. Biochem. _5_72505-518;
`Beutler, B. and Cerami, A. (1989) Annu. Rev. Immunol. 1:625-655). Furthermore,
`
`TNFOC has been implicated in the pathophysiology of a variety of other human diseases
`
`15
`
`and disorders, including sepsis, infections, autoimmune diseases, transplant rejection
`
`and graft-versus-host disease (see e. g., Moeller, A., et al. (1990) Cytokine _2_:162-169;
`U.S. Patent No. 5,231,024 to Moeller et al.; European Patent Publication No. 260 610
`
`B1 by Moeller, A., et al.Vasilli, P. (1992) Annu. Rev. Immunol. 192411-452; Tracey,
`
`K]. and Cerami, A. (1994) Annu. Rev. Med. 452491-503).
`
`20
`
`Because of the harmful role of human TNFOL (hTNFoc) in a variety of human
`
`disorders, therapeutic strategies have been designed to inhibit or counteract hTNFOL
`activity. In particular, antibodies that bind to, and neutralize, hTNFoc have been sought
`as a means to inhibit hTNFoc activity. Some of the earliest of such antibodies were
`
`mouse monoclonal antibodies (mAbs), secreted by hybridomas prepared from
`
`25
`
`lymphocytes of mice immunized with hTNFOc (see e.g., Hahn T; et al., (1985) Proc Natl
`Acad Sci USA 82: 3814-3818; Liang, C-M., er al. (1986) Biochem. Biophys. Res.
`
`Commun. lfl: 847-854; Hirai, M., et al. (1987) J. Immunol. Methods _9_6_:57-62; Fendly,
`
`B.M., et al. (1987) Hybridoma §:359—370; Moeller, A., et al. (1990) Cytokine _2_.:162-
`
`169; U.S. Patent No. 5,231,024 to Moeller et al.; European Patent Publication No. 186
`
`833 B1 by Wallach, D.; European Patent Application Publication No. 218 868 A1 by
`Old ez.‘ al.; European Patent Publication No. 260 610 B1 by Moeller, A., et al.). While
`these mouse anti-hTNFoc antibodies often displayed high affinity for hTNFOc (e.g., Kd S
`
`10'9M) and were able to neutralize hTNFoL activity, their use in vivo may be limited by
`problems associated with administration of mouse antibodies to humans, such as short
`serum half life, an inability to trigger certain human effector functions and elicitation of
`an unwanted immune response against the mouse antibody in a human (the "human anti-
`
`30
`
`35
`
`mouse antibody" (HAMA) reaction).
`
`Ex. 1047 — Page 2 of 46
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`WO 2004/016286
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`-2-
`
`In an attempt to overcome the problems associated with use of fully-murine
`
`antibodies in humans, murine anti-hTNFoc antibodies have been genetically engineered
`
`to be more "human—like." For example, chimeric antibodies, in which the variable
`
`regions of the antibody chains are murine-derived and the constant regions of the
`antibody chains are human—derived, have been prepared (Knight, D.M, et al. (1993)
`Mol. Immunol. $:1443—1453; PCT Publication No. W0 92/ 16553 by Daddona, P.E., et
`
`al.). Additionally, humanized antibodies, in which the hypervariable domains of the
`antibody variable regions are murine-derived but the remainder of the variable regions
`and the antibody constant regions are human-derived, have also been prepared (PCT
`
`10
`
`Publication No. W0 92/ 1 1383 by Adair, J.R., et al.). However, because these chimeric
`
`and humanized antibodies still retain some murine sequences, they still may elicit an
`
`unwanted immune reaction, the human anti-chimeric antibody (HACA) reaction,
`
`especially when administered for prolonged periods, e.g. , for chronic indications, such
`as rheumatoid arthritis (see e. g., Elliott, M.J., et al. (1994) Lancet fiflfiz 1 125-1127; Elliot,
`
`15
`
`M.J., et al. (1994) Lancet 3_4l4l_:11O5-1110).
`
`A preferred hTNFOc inhibitory agent to murine mAbs or derivatives thereof (e.g. ,
`chimeric or humanized antibodies) would be an entirely human anti-hTNFOc antibody,
`
`since such an agent should not elicit the HAMA reaction, even if used for prolonged
`
`20
`
`periods. Human monoclonal autoantibodies against hTNFOL have been prepared using
`human hybridoma techniques (Boyle, P., et al. (1993) Cell. Immunol. 1'5_2:556—568;
`Boyle, P., et al. (1993) Cell. Immunol. 1_5.2_:569-581; European Patent Application
`Publication No. 614 984 A2 by Boyle, et al.). However, these hybridoma-derived
`
`monoclonal autoantibodies were reported to have an affinity for hTNFOc that was too
`
`low to calculate by conventional methods, were unable to bind soluble hTNFOL and were
`
`25
`
`unable to neutralize hTNFoc—induced cytotoxicity (see Boyle, et al.; supra). Moreover,
`
`the success of the human hybridoma technique depends upon the natural presence in
`
`human peripheral blood of lymphocytes producing autoantibodies specific for hTNFoc.
`Certain studies have detected serum autoantibodies against hTNFOt in human subjects
`
`(Fomsgaard, A., et al. (1989) Scand. J. Immunol. §Q:219-223; Bendtzen, K., et al.
`(1990) Prog. Leukocyte Biol. 19_B:447-452), whereas others have not (Leusch, H-G., et
`
`30
`
`al. (1991) J. Immunol. Methods _1_fl:145—l47).
`
`Alternative to naturally—occurring human anti-hTNFot antibodies would be a
`
`recombinant hTNFOL antibody. Recombinant human antibodies that bind hTNFot with
`relatively low affinity (i.e., Kd ~10'7M) and a fast off rate (i. e., Koff ~ 10'2 sec‘1) have
`been described (Griffiths, A.D., et al. (1993) EMBO J. ;g:725—734). However, because
`
`35
`
`of their relatively fast dissociation kinetics, these antibodies may not be suitable for
`therapeutic use. Additionally, a recombinant human anti—hTNFoc has been described
`
`Ex. 1047 — Page 3 of 46
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`-3-
`
`that does not neutralize hTNFot activity, but rather enhances binding of hTNFOL to the
`
`surface of cells and enhances internalization of hTNFoc (Lidbury, A., et al. (1994)
`
`Biotechnol. Ther. _5_:27-45; PCT Publication No. WO 92/03145 by Aston, R. et al.)
`
`Recombinant human antibodies that bind soluble hTNFoc with high affinity and
`
`slow dissociation kinetics and that have the capacity to neutralize hTNFOc activity,
`
`including hTNFoc-induced cytotoxicity (in vitro and in vivo) and hTNFoc—induced cell
`
`activation, have also been described (see U.S. Patent No. 6,090,382).
`
`Summary of the Invention
`
`10
`
`There is a need for a stable aqueous pharmaceutical formulation with an
`
`extended shelf life, comprising an antibody which is suitable for therapeutic use to
`
`inhibit or counteract detrimental hTNFoc activity. There is also a need for a stable
`
`aqueous pharmaceutical formulation with an extended shelf life, comprising an antibody
`suitable for therapeutic use which is easily administered and contains a high protein
`
`15
`
`concentration.
`
`This invention provides a liquid aqueous pharmaceutical formulation consisting
`
`of a therapeutically effective amount of an antibody in a buffered solution forming a
`
`formulation having a pH between about 4 and about 8 and having a shelf life of at least
`
`20
`
`18 months. The invention also includes an aqueous pharmaceutical formulation
`
`comprising a therapeutically effective amount of an antibody in a buffered solution
`
`forming a formulation having a pH between about 4 and 8 and having a shelf life of at
`
`least 18 months in the liquid state. In one embodiment, the pharmaceutical formulation
`
`has enhanced stability. In a further embodiment, the formulation of the invention is
`
`25
`
`stable following at least 3 freeze/thaw cycles of the formulation. In another
`
`embodiment, the antibody is directed to TNFOL. In yet another embodiment, the
`
`antibody is directed to human TNFOL. In still another embodiment, the antibody is
`
`D2E7.
`
`This invention also provides a liquid aqueous pharmaceutical formulation
`
`30
`
`comprising a therapeutically effective amount of an antibody in a buffered solution
`
`forming a formulation having a pH between 4 and 8 and having enhanced stability of at
`
`least 12 months at a temperature of 2 - 8°C. In one embodiment, the formulation has
`
`enhanced stability of at least 18 months. In another embodiment, the antibody is
`directed to TNFOL.
`In yet another embodiment, the antibody is directed to human
`
`35
`
`TNFOL. In a further embodiment, the antibody is DZE7.
`
`The invention further provides a liquid aqueous pharmaceutical formulation
`
`comprising a therapeutically effective amount of an antibody in a buffered solution
`
`Ex. 1047 — Page 4 of 46
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`

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`WO 2004/016286
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`PCT/IB2003/004502
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`_4_
`
`forming a formulation having a pH between about 4 and about 8 which is easily
`administratable. In one embodiment, the antibody is directed to TNFOL.
`In yet another
`
`embodiment, the antibody is directed to human TNFOL. In a further embodiment, the
`
`antibody is DZE7.
`
`In one embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation is suitable for injection. In a further embodiment, the formulation is
`
`suitable for single use so injection. In another embodiment, the concentration of the
`
`antibody in the liquid aqueous pharmaceutical formulation is about 1-150 mg/ml. In yet
`another embodiment, the concentration of the antibody in the formulation is about 50
`
`10
`
`mg/ml. In still another embodiment, the formulation has a high protein concentration.
`
`In yet another embodiment of the invention, the formulation is not light sensitive.
`In one embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation contains an antibody, or an antigen-binding portion thereof, that dissociates
`from human TNFOC with a Kd of 1 x 108 M or less and a Koff rate constant of 1 x 10'3 s‘
`
`15
`
`1 or less, both determined by surface plasmon resonance, and neutralizes human TNFOC
`cytotoxicity in a standard in vitro L929 assay with an IC50 of 1 X 107 M or less.
`In
`another embodiment, the formulation of the invention contains an antibody, or antigen—
`
`binding portion thereof, which dissociates from human TNFOC with a Koff rate constant
`of 5 x 1O'4 s‘1 or less. In a further embodiment, the formulation contains an antibody,
`
`20
`
`or antigen-binding portion thereof, which dissociates from human TNFOL with a Koff rate
`
`constant of 1 x 104 s'1 or less. In still a further embodiment, the formulation of the
`
`invention contains an antibody, or antigen-binding portion thereof, which neutralizes
`human TNFOL cytotoxicity in a standard in vitro L929 assay with an IC50 of 1 x 1O'8 M
`
`or less. In yet another embodiment of the invention, the claimed formulation includes
`
`25
`
`an antibody, or antigen-binding portion thereof, which neutralizes human TNFOC
`cytotoxicity in a standard in vitro L929 assay with an IC50 of 1 x 1O'9I M or less.
`Another embodiment of the invention, includes a formulation where the antibody, or
`antigen-binding portion thereof, neutralizes human TNFOL cytotoxicity in a standard in
`vitro L929 assay with an IC50 of 1 x 1040 M or less.
`
`30
`
`In another embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation contains of an antibody, or antigen-binding portion thereof, which is a
`
`recombinant antibody, or antigen-binding portion thereof. In another embodiment, the
`
`formulation contains an antibody, or antigen-binding portion thereof, which inhibits
`
`human TNFOL-induced expression of ELAM-1 on human umbilical vein endothelial
`
`35
`
`cells. In still another embodiment, the claimed formulation includes the DZE7 antibody.
`
`In another embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation includes an antibody, or antigen-binding portion, thereof which dissociates
`
`Ex. 1047 — Page 5 of 46
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`-5-
`
`from human TNFOL with a Koff rate constant of 1 x 103 s‘1 or less, as determined by
`
`surface plasmon resonance;
`
`b) has a light chain CDR3 domain comprising the amino acid sequence of SEQ
`
`ID NO: 3, or modified from SEQ ID NO: 3 by a single alanine substitution at position 1,
`
`4, 5, 7 or 8 or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6,
`
`7, 8 and/or 9;
`
`c) has a heavy chain CDR3 domain comprising the amino acid sequence of SEQ
`
`ID NO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution at position 2,
`
`3, 4, 5, 6, 8, 9, 10 or 11 or by one to five conservative amino acid substitutions at
`
`10
`
`another embodiment, the formulation of
`positions 2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12.
`the invention includes an antibody, or an antigen—binding portion thereof, which
`dissociates from human TNFOL with a Koff rate constant of 5 x 10‘4 s'1 or less. In yet
`
`another embodiment of the invention, the formulation includes an antibody, or an
`
`antigen-binding portion thereof, which dissociates from human TNFoc with a Koff rate
`constant of 1 X 10'4 s'1 or less.
`
`15
`
`In yet another embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation, contains of an antibody, or antigen-binding portion thereof, which has a
`
`light chain variable region (LCVR) having a CDR3 domain comprising the amino acid
`sequence of SEQ ID NO: 3, or modified from SEQ ID NO: 3-by a single alanine
`substitution at position 1, 4, 5, 7 or 8, and with a heavy chain variable region (HCVR)
`
`20
`
`having a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, or
`
`modified from SEQ ID NO: 4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8,
`
`9, 10 or 11. In a further embodiment, the formulation of the invention contains an
`
`antibody, wherein the LCVR of the antibody, or an antigen-binding portion thereof,
`
`25
`
`further has a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5 and
`
`the HCVR of the antibody, or an antigen-binding portion thereof, further has a CDR2
`
`domain comprising the amino acid sequence of SEQ ID NO: 6. In yet another
`
`embodiment, the formulation of the invention contains an antibody, wherein the LCVR
`
`of the antibody, or an antigen-binding portion thereof, further has CDR1 domain
`
`130
`
`comprising the amino acid sequence of SEQ ID NO: 7 and the HCVR has a CDR1
`
`domain comprising the amino acid sequence of SEQ ID NO: 8.
`
`In yet another embodiment of the invention, the antibody or antigen-binding
`
`portion thereof, contained in the liquid aqueous pharmaceutical formulation has a light
`chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO? 1
`
`35
`
`and a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ
`
`ID NO: 2. In another embodiment, the antibody, or antigen-binding portion thereof, has
`
`an IgG1 heavy chain constant region. In still another embodiment, the antibody, or
`
`Ex. 1047 — Page 6 of 46
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`-6-
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`antigen—binding portion thereof, has an IgG4 heavy chain constant region. In another
`
`embodiment, the antibody, or antigen—binding portion thereof, is a Fab fragment. In still
`
`a further embodiment, the antibody, or antigen—binding portion thereof, is a single chain
`
`Ev fragment.
`
`In one embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation, contains an antibody, or antigen—binding portion thereof, which has a light
`chain variable region (LCVR) having a CDR3 domain comprising an amino acid
`sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 11, SEQ ID
`NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID
`
`10
`
`NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID
`
`NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26 or with a
`
`heavy chain variable region (HCVR) having a CDR3 domain comprising an amino acid
`
`sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 27, SEQ ID
`NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID
`
`15
`
`NO: 33 and SEQ ID NO: 34. In still another embodiment, the antibody, or antigen-
`
`binding portion thereof, neutralizes the activity of human TNFOL, chimpanzee TNFOC and
`
`at least one additional primate TNFOL selected from the group consisting of baboon
`
`TNFOC, marrnoset TNFOL, cynomolgus TNFOL and rhesus TNFOL. In a further
`
`embodiment, the formulation of the invention includes an antibody, or an antigen-
`
`20
`
`binding portion thereof, which also neutralizes the activity of mouse TNFOC. The
`
`formulation of the invention also an antibody, or an antigen—binding portion thereof,
`
`which neutralizes the activity of pig TNFOL.
`
`In a further embodiment, the invention provides a liquid aqueous pharmaceutical
`formulation which contains an antibody, or antigen—binding portion thereof, which binds
`
`25
`
`to human TNFOC and comprises:
`
`a light chain CDR3 domain comprising the amino acid sequence of SEQ ID NO:
`
`3, or modified from SEQ ID NO: 3 by a single alanine substitution at position 1, 4, 5, 7
`
`or 8 or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6, 7, 8
`
`and/or 9, and
`
`30
`
`a heavy chain CDR3 domain comprising the amino acid sequence of SEQ ID
`
`NO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution at position 2, 3,
`
`4, 5, 6, 8, 9, 10 or 11 or by one to five conservative amino acid substitutions at positions
`
`2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12. In one embodiment, the liquid aqueous
`
`pharmaceutical formulation includes an antibody which bind human TNFOL and
`comprises a light chain variable region (LCVR) having a CDR3 domain comprising an
`
`35
`
`amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO:
`
`11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:
`
`Ex. 1047 — Page 7 of 46
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`
`16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO:
`
`21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ DD NO: 24, SEQ ID NO: 25, SEQ ID NO:
`
`26 or a heavy chain variable region (HCVR) having a CDR3 domain comprising an
`
`amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO:
`
`27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO:
`
`32, SEQ ID NO: 33 and SEQ ID NO: 34. In a further embodiment of the invention, the
`
`antibody, or antigen-binding portion thereof, binds human TNFOL and is the antibody
`
`D2E7 or an antigen binding portion thereof.
`
`The invention also provides an aqueous pharmaceutical composition comprising
`
`10
`
`a polyol, a surfactant, and a buffer system comprising citrate and/or phosphate with a pH
`
`of about 4 to 8, in amounts sufficient to formulate an antibody for therapeutic use at a
`
`concentration of greater than about 45 mg/ml. In one embodiment, the polyol is
`
`mannitol and the surfactant is polysorbate 80. In another embodiment, the composition
`
`includes 5-20 mg/ml of mannitol and 0.1-10 mg/ml of polysorbate 80. In a further
`
`15
`
`embodiment, the composition includes the antibody D2E7.
`
`The invention also provides a liquid aqueous pharmaceutical formulation
`
`consisting of 1-150 mg/ml of antibody, 5-20 mg/ml of mannitol, 0.1-10 mg/ml of
`
`Tween-80, and a buffer system comprising citrate and/or phosphate, with a pH of 4 to 8.
`
`In one embodiment, the antibody is directed to hTN1-“tot. In another embodiment, the
`
`20
`
`formulation contains about 40 mg of antibody. The invention further provides a liquid
`
`aqueous pharmaceutical formulation comprising about 50 mg/ml of antibody, about 12
`
`mg/ml of mannitol, about 1 mg/ml of Tween-80, and a buffer system comprising citrate
`
`and/or phosphate, with a pH of about 4 to about 8. In one embodiment, the pH of the
`
`formulation is between about 4.5 to about 6.0. In another embodiment, the pH is
`
`25
`
`between about 4.8 to about 5.5. In yet another embodiment, the pH of the invention
`
`is between about 5.0 to about 5.2.
`
`In one embodiment of the invention, the liquid aqueous pharmaceutical
`
`formulation also includes about 1.305 mg/ml of citric acid, about 0.305 mg/ml of
`
`sodium citrate, about 1.53 mg/ml of disodium phosphate dihydrate, about 0.86 mg/ml of
`
`30
`
`sodium dihydrogen phosphate dihydrate, and about 6.165 mg/ml of sodium chloride. In
`
`another embodiment, the formulation of the invention includes an antibody which is
`
`directed to hTNFoc. In yet another embodiment, the formulation of the invention
`
`includes the antibody D2E7.
`
`In yet a further embodiment, the formulation of the
`
`invention is administered to a subject suffering from a disorder in which TNFOL activity
`
`35
`
`is detrimental such that TNFOL activity in the subject is inhibited
`
`Ex. 1047 — Page 8 of 46
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`Ex. 1047 - Page 8 of 46
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`WO 2004/016286
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`PCT/IB2003/004502
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`Detailed Description of the Invention
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`This invention pertains to a liquid aqueous pharmaceutical formulation with a pH
`of about 4 to about 8 which contains a high protein concentration, including an antibody
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`concentration ranging from about 1 to about 150 mg/ml, and has enhanced stability.
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`This invention also pertains to a liquid aqueous pharmaceutical formulation for
`therapeutic use in a subject suffering from a condition characterized by detrimental
`TNFOL activity. The formulation of the invention comprises the following constituents:
`an antibody which binds to human TNFOL with high affinity, a low off rate and high
`neutralizing capacity; a buffer, which includes citric acid, sodium citrate, disodium
`phosphate dihydrate, and sodium dihydrogen phosphate dihydrate; tonicity agents,
`which include mannitol and sodium chloride; a detergent, including polysorbate 80; and
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`sodium hydroxide, for pH adjustment.
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`Definitions
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`In order that the present invention may be more readily understood, certain terms
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`are first defined.
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`The term “subject” is intended to include living organisms, e.g., prokaryotes and
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`eukaryotes. Examples of subjects include mammals, e.g., humans, dogs, cows, horses,
`pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non-human animals. In
`specific embodiments of the invention, the subject is a human.
`A1
`The term "pharmaceutical formulation" refers to preparations which are in such
`form as to permit the biological activity of the active ingredients to be unequivocally
`effective, and which contain no additional components which are significantly toxic to
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`the subjects to which the formulation would be administered. "Pharmaceutically
`acceptable" excipients (vehicles, additives) are those which can reasonably be
`administered to a subject mammal to provide an effective dose of the active ingredient
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`employed.
`A "stable" formulation is one in which the antibody therein essentially retains its
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`physical stability and/or chemical stability and/or biological activity upon storage.
`Various analytical techniques for measuring protein stability are available in the art and
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`are reviewed in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel
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`Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:
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`29-90 (1993), for example. Stability can be measured at a selected temperature for a
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`selected time period. Preferably, the formulation is stable at room temperature (about
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`30°C) or at 40°C for at least 1 month and/or stable at about 2—8°C. for at least 1 year for
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`Ex. 1047 — Page 9 of 46
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`Ex. 1047 - Page 9 of 46
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`WO 2004/016286
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`PCT/IB2003/004502
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`at least 2 years. Furthermore, the formulation is preferably stable following freezing (to,
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`e.g., —70°C) and thawing of the formulation, hereinafter referred to as a "freeze/thaw
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`cycle."
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`An antibody "retains its physical stability" in a pharmaceutical formulation if it
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`shows substantially no signs of aggregation, precipitation and/or denaturation upon
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`visual examination of color and/or clarity, or as measured by UV light scattering or by
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`size exclusion chromatography.
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`An antibody "retains its chemical stability" in a pharmaceutical formulation, if
`the chemical stability at a given time is such that the antibody is considered to still retain
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`its biological activity as defined below. Chemical stability can be assessed by detecting
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`and quantifying chemically altered forms of the antibody. Chemical alteration may
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`involve size modification (e. g. clipping) which can be evaluated using size exclusion
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`chromatography, SDS—PAGE and/or matrix-assisted laser desorption ionization/time-of-
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`flight mass spectrometry (MALDI/TOF MS), for example. Other types of chemical
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`alteration include charge alteration (e. g. occurring as a result of deamidation) which can
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`be evaluated by ion-exchange chromatography, for example.
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`An antibody "retains its biological activity" in a pharmaceutical formulation, if
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`the antibody in a pharmaceutical formulation is biologically active for its intended
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`purpose. For example, biological activity is retained if the biological activity of the
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`antibody in the pharmaceutical formulation is within about 30%, about 20%, or about
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`10% (within the errors of the assay) of the biological activity exhibited at the time the
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`pharmaceutical formulation was prepared (e. g., as determined in an antigen binding
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`assay).
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`"Isotonic" is a term recognized in the art. Isotonic can mean, for example, that
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`the formulation of interest has essentially the same osmotic pressure as human blood.
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`Isotonic formulations will generally have an osmotic pressure from about 250 to 350
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`mOsm. Isotonicity can be measured using a vapor pressure or ice-freezing type
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`osmometer, for example. A "tonicity agent" is a compound which renders the
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`formulation isotonic.
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`A "polyol" is a substance with multiple hydroxyl groups, and includes sugars
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`(reducing and nonreducing sugars), sugar alcohols and sugar acids. Preferred polyols
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`herein have a molecular weight which is less than about 600 kD (e. g. in the range from
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`about 120 to about 400 kD). A "reducing sugar" is one which contains a hemiacetal
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`group that can reduce metal ions or react covalently with lysine and other amino groups
`in proteins and a "nonreducing sugar" is one which does not have these properties ofa
`reducing sugar. Examples of reducing sugars are fructose, mannose, maltose, lactose,
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`arabinose, xylose, ribose, rhamnose, galactose and glucose. Nonreducing sugars include
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`EX. 1047 - Page 10 of 46
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`Ex. 1047 - Page 10 of 46
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`sucrose, trehalose, sorbose, melezitose and raffinose. Mannitol, xylitol, erythritol,
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`threitol, sorbitol and glycerol are examples of sugar alcohols. As to sugar acids, these
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`include L-gluconate and metallic salts thereof. Where it desired that the formulation is
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`freeze—thaw stable, the polyol is preferably one which does not crystallize at freezing
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`temperatures (e.g. —20°C) such that it destabilizes the antibody in the formulation. The
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`polyl may also act as a tonicity agent. In one embodiment of the invention, one
`ingredient of the formulation is mannitol in a concentration of 5 to 20 mg/ml. In a
`preferred embodiment of the invention, the concentration of mannitol is 7.5 to 15
`mg/ml. In a more preferred embodiment of the invention, the concentration of mannitol
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`is 10-14 mg/ml.
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`As used herein, "buffer" refers to a buffered solution that resists changes in pH
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`by the action of its acid-base conjugate components. The buffer of this invention has a
`pH in the range from about 4 to about 8; preferably from about 4.5 to about 7; and most
`preferably has a pH in the range from about 5.0 to about 6.5. Examples of buffers that
`will control the pH in this range include acetate (e.g. sodium acetate), succinate (such as
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`sodium succinate), gluconate, histidine, citrate and other organic acid buffers.
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`In a pharmacological sense, in the context of the present invention, a
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`"therapeutically effective amount" or "effective amount" of an antibody refers to an
`amount effective in the prevention or treatment of a disorder for the treatment of which
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`the antibody is effective. A "disorder" is any condition that would benefit from
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`treatment with the antibody. This includes chronic and acute disorders or diseases
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`including those pathological conditions which predisposes the subject to the disorder in
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`question.
`A "preservative" is a compound which can be included in the formulation to
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`essentially reduce bacterial action therein, thus facilitating the production of a multi-use
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`formulation, for example. Examples of potential preservatives include
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`octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium
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`chloride (a mixture of alkylbenzyldimethylammonium chlorides in which the alkyl
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`groups are long—chain compounds), and benzethonium chloride. Other types of
`preservatives include aromatic alcohols such as phenol, butyl and benzyl alcohol, alkyl
`parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3~
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`pentanol, and m-cresol.
`"Treatment" refers to both therapeutic treatment and prophylactic or preventative
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`measures. Those in need of treatment include those already with the disorder as well as
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`those in which the disorder is to be prevented.
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`The phrases "parenteral administration" and "administered parenterally" as used
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`herein means modes of administration other than enteral and topical administration,
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`EX. 1047 - Page 11 of 46
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`Ex. 1047 - Page 11 of 46
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`WO 2004/016286
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`PCT/IB2003/004502
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`usually by injection, and includes, without limitation, intravenous, intramuscular,
`intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intraderrnal,
`intraperitoneal, transtrachcal, subcutaneous, subcuticular, intraarticular, subcapsular,
`subarachnoid, intraspinal and intrastemal injection and infusion.
`The phrases "systemic administration," "administered systemically," "peripheral
`administration" and "administered peripherally" as used herein mean the administration
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`of a compound, drug or other material other than directly into the central nervous
`system, such that it enters the patient's system and, thus, is subject to metabolism and
`other like processes, for example, subcutaneous administration.
`The phrase "pharmaceutically acceptable carrier" is art recognized and includes a
`pharmaceutically acceptable material, composition or vehicle, suitable for
`admi