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`I 1111111111111111 11111 1111111111 1111111111 11111 1111111111111111 IIII IIII IIII
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`
`US009487581B2
`
`(IO) Patent No.:
`
`US 9,487,581 B2
`
`(12)United States Patent
`(45)Date of Patent:
`Nov. 8, 2016
`
`
`Abate et al.
`
`Kakuta et al.
`2005/0214278 Al
`
`9/2005
`(54)ANTI-CTLA-4
`ANTIBODY COMPOSITIONS
`
`2006/0008415 Al*
`1/2006
`
`Kaisheva et al. ............ 424/1.49
`
`2006/0088523 Al*
`4/2006
`
`Andya et al. .............. 424/133.l
`(75)Inventors: Abate, Chesterfield, MO (US);
`
`
`Justin
`Yang et al.
`
`2006/0182740 Al
`8/2006
`
`Chesterfield, MO
`Kevin Muthurania,
`
`2007 /0048332 Al *
`3/2007
`
`Oliver et al. .............. 424/204.l
`
`
`(US); Sandeep Nema, Chesterfield, MO
`
`Das et al. .................. 424/142.1
`
`2008/0248047 Al *
`10/2008
`
`(US); Satish Singh, Chesterfield, MO
`
`2008/0292639 Al *
`
`
`11/2008 Shen et al .................. 424/158.1
`
`2009/0110681 Al*
`4/2009
`
`
`Carroll ............... A61K 39/3955
`
`
`(US); Carrie Seattle, WA (US);
`Elliott,
`424/139.1
`
`Tapan Das, Chesterfield, MO (US)
`2009/0117103 Al * 5/2009
`
`
`
`Devalaraja et al. ....... 424/133.l
`
`2010/0119517 Al* 5/2010
`
`
`Burgess ..................... 424/141.1
`
`New York, NY (US)
`(73)Assignee:
`Pfizer Inc.,
`
`(21)Appl. No.:11/817,894
`
`(22)PCT Filed:Mar. 2, 2006
`
`( *) Notice: Subject to any disclaimer, the term of this
`
`
`
`
`
`patent is extended or adjusted under 35
`AR
`AR
`
`
`U.S.C. 154(b) by 1866 days.
`AR
`CA
`EP
`EP
`JP
`JP
`(86)PCT No.: PCT/US2006/007555
`KR
`RU
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`
`§371 (c)(l),
`(2), ( 4) Date: Apr. 18, 2008
`
`
`
`(87)PCT Pub. No.: WO2006/096491
`
`
`
`PCT Pub. Date: Sep. 14, 2006
`
`(65)
`
`
`
`Prior Publication Data
`
`
`
`US 2009/0130119 Al May 21, 2009
`
`
`
`
`
`Related U.S. Application Data
`
`FOREIGN PATENT DOCUMENTS
`
`3/2006
`P 06 01 00793
`
`P 06 01 00794 3/2006
`
`P 06 01 00796 3/2006
`
`2600608 9/2006
`
`
`0612251 Bl 12/1997
`
`1262193 12/2002
`
`2002-537226 11/2002
`
`2006-249085 9/2006
`10/2004
`10-2004-0085185
`
`1438240 C
`3/1996
`WO 97/45140
`12/1997
`WO 00/37504
`6/2000
`
`WO0l/14424 A2
`3/2001
`
`WO03/039485 A2
`5/2003
`
`W02004/007520 A2
`1/2004
`W02004/091658
`10/2004
`WO2006/044908
`4/2006
`WO 2006/096461
`9/2006
`WO 2006/096488
`9/2006
`WO 2006/096490
`9/2006
`
`OTHER PUBLICATIONS
`
`(51)Int. Cl.
`A61K 39/395
`
`(2006.01)
`(2006.01)
`(2006.01)
`
`Rudikoff et al., Proc Natl Acad Sci USA 79: 1979-1983, 1982.*
`
`
`
`
`
`
`(60)Provisional application No. 60/659,766, filed on Mar.
`
`
`
`Kussie et al., J. Immunol. 152: 146-152, 1994.*
`
`
`
`8, 2005, provisional application No. 60/728, 165, filed
`
`Chen et al., EMBO J., 14: 2784-2794, 1995.*
`
`
`on Oct. 19, 2005, provisional application No.
`
`
`
`
`Colman, Research in Immunology 145: 33-36, 1994.*
`
`60/752,712, filed on Dec. 20, 2005, provisional
`
`
`
`
`Hodi,F., et al., "Biologic Activity of Cytotoxic T Lymphocyte­
`
`
`application No. 60/762,456, filed on Jan. 26, 2006.
`
`
`
`Associated Antigen 4 Antibody Blockade in Previously
`
`
`Vaccina ... "PNAS, Apr. 15, 2003, 4712-4717, vol. 100, No. 8.
`
`
`
`
`Pistillo, M., et al, :Molecular Characterization and Applications of
`
`
`Recombinant scFv Antibodies to CD152 Co-Stimulatory
`C07K 16124
`
`
`
`Molecu . . . Tissue Antigens, Mar. 2000, 229-238, vol. 55.
`A61K 39/00
`
`
`
`
`Daugherty, Al, et al, "Formulation and Delivery Issues for Mono­
`(52)U.S. Cl.
`
`
`
`clonal Antibody Therapeutics," Advanced Drug Delivery Reviews,
`
`CPC ......... C07K 16/243 (2013.01);
`
`2006, 686-706, 58.
`A61K 39/3955
`
`A61K 39/39591 A61K
`
`(2013.01); (2013.01);
`Chen, B., et al, "Influence of Histidine on the Stability and Physical
`
`
`
`
`2039/505 C07K 2317/21
`
`(2013.01); (2013.01)
`
`
`
`Properties of a Fully Human . . . "Pharmaceutical Research, Dec.
`
`2003, 1952-1960, vol. 20, No. 12.
`( 58)Field of Classification
`Search
`Wang, "Instability, stabilization, and formulation of liquid protein
`
`
`
`
`None
`
`International Journal of Pharmaceutics
`pharmaceuticals,"
`185
`
`See application file for complete search history.
`
`(2):129-188 (1999).
`(Continued)
`
`
`
`
`
`(56)
`
`
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`et al. 4,597,966 A 7/1986 Zolton
`
`
`
`5,654,403 A 8/1997 Smith et al.
`
`5,792,838 A 8/1998 Smith et al.
`
`
`5,804,557 A 9/1998 Cleland et al.
`
`6,171,586 Bl 1/2001 Lam et al.
`
`6,267,958 Bl 7/2001 Andya et al.
`
`6,682,736 Bl 1/2004 Hanson et al.
`
`2003/0118583 A l 6/2003 Emery et al.
`
`
`2003/0138417 Al 7/2003 Kaisheva et al.
`
`2003/0190316 Al 10/2003
`Kakuta et al.
`
`
`et al. 2004/0038878 Al 2/2004 Tanikawa
`
`2004/0057951 Al 3/2004 Bednar et al.
`
`Primary Examiner
`
`-Phillip Gambel
`
`
`(74)Attorney, Agent, or Firm -Ropes & Gray LLP
`
`(57)
`
`ABSTRACT
`
`The present invention provides for novel compositions of
`
`
`
`
`
`
`
`
`
`anti-CTLA-4 antibodies comprising a chelating agent. Also
`
`
`
`
`
`provided are method of treating diseases and conditions with
`
`
`
`novel compositions of CTLA-4 antibodies, including vari­
`
`ous neoplasia conditions.
`
`
`
`20 Claims, 12 Drawing Sheets
`
`
`
`TRANSGENE/BIOINVENT
`EXHIBIT 1071
`
`Page 1 of 73
`
`

`

`US 9,487,581 B2
`Page 2
`
`(56)
`
`References Cited
`
`OTHER PUBLICATIONS
`
`Camacho et al., “Novel therapies targeting the immune system:
`CTLA4 blockade with tremelimumab (CP-675,206), a fully human
`monoclonal antibody,” Expert Opin. Investig. Drugs 17(3):37 1-385
`(2008).
`Canniff et al., “CP-675,205 anti-CTLA4antibody clinical candidate
`enhances IL-2 production in cancer patient T cells in vitro regard-
`less of tumortype or stage of disease,” Proc Am Assoc Cancer Res
`45: Abstract 709 (2004).
`Cranmeret al., “The Role of the CTLA4 Blockadein the Treatment
`of Malignant Melanoma,” Cancer Investigation 25:613-631 (2007).
`Hamilton et al., “Human IgG Subclass Measurements in the Clinical
`Laboratory,” Clin. Chem. 33(10):1707-1725 (1987).
`
`O’Day et al., “Targeting Cytotoxic T-Lymphocyte Antigen-4
`(CTLA-4): A Novel Strategy for the Treatment of Melanoma and
`Other Malignancies,” Cancer 110(12):2614-2627 (2007).
`Perchiaccaetal., “Engineering Aggregation-Resistant Antibodies,”
`Annu, Rey. Chem. Biomol. Eng. 3:263-286 (2012).
`Frokjaer and Hovgaard, “Pharmaceutical Formulation Develop-
`ment ofPeptides and Proteins,” Taylor & Francis Limited (2000).
`Tsai et al., “Formulation design of acidic fibroblast growth factor,”
`Pharm Research 10(5): 649-659 (1993).
`Ruiz et al., “Long-term stabilization of recombinant human inter-
`feron alpha 2b in aqueous solution without serum albumin,” Jnt.
`Journal ofPharmaceutics 264:57-72 (2003).
`Wang et al., “Antibody structure, instability, and formulation,” J
`Pharm Sci. 96(1):1-26 (2007).
`
`* cited by examiner
`
`Page 2 of 73
`
`Page 2 of 73
`
`

`

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`
`U.S. Patent
`
`Nov.8, 2016
`
`Sheet 1 of 12
`
`US 9,487,581 B2
`
`
`
`
`
`
`
`PercentAggregationVersusTimeforFormulationsfromExample4UnderStorageat40°C.
`
`
`
`
`
`Page 3 of 73
`
`

`

`U.S. Patent
`
`Nov.8, 2016
`
`Sheet 2 of 12
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`percent
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`PCEOTETECLOTT
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`US 9,487,581 B2
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`SN 8
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`PercentFragmentation(hydrolysisimpurities)VersusTimeforFormulationsfromExample4UnderStorageat40°C.
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`Page 4 of 73
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`Page 4 of 73
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`

`

`Nov.8, 2016
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`Sheet 3 of 12
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`US 9,487,581 B2
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`U.S. Patent
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`Nov.8, 2016
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`Sheet 4 of 12
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`Page 6 of 73
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`U.S. Patent
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`Nov.8, 2016
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`Sheet 5 of 12
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`US 9,487,581 B2
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`U.S. Patent
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`Sheet 6 of 12
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`Page 8 of 73
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`U.S. Patent
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`Nov.8, 2016
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`Sheet 7 of 12
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`US 9,487,581 B2
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`Page 9 of 73
`
`

`

`U.S. Patent
`
`Nov.8, 2016
`
`Sheet 8 of 12
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`US 9,487,581 B2
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`Page 10 of 73
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`

`

`Nov.8, 2016
`
`Sheet 9 of 12
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`US 9,487,581 B2
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`Page 11 of 73
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`

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`U.S. Patent
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`Nov.8, 2016
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`Sheet 10 of 12
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`Page 12 of 73
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`Page 12 of 73
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`

`

`U.S. Patent
`
`Nov.8, 2016
`
`Sheet 11 of 12
`
`US 9,487,581 B2
`
`FIG. 11A
`
`Ticilimumab (11.2.1) Heavy Chain DNA (SEQ ID NO:1)
`
`atggagtttg ggctgagctg ggttttccte gttgctcttt taagaggtgt ccagtatcag
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`
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`
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`
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`
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`
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`
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`
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`
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`
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`
`660
`
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`
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`aaggacaccc
`
`teatgatcte
`
`ecogacecct
`
`gaggtcacgt
`
`gegtggtggt ggacgtgagc
`
`cacgaagacc
`
`ecgaggtcca
`
`gttcaactgg
`
`tacgtggacg
`
`gegtggaggt gcataatgec
`
`aagacaaagc
`
`cacgagagga
`
`gcagttcaac
`
`780
`
`840
`
`900
`
`960
`
`agcacgttcc
`
`gtgtggtcag egtcctcacc
`
`gttgtgcacc
`
`aggactggct
`
`gaacggcaag
`
`1020
`
`gagtacaagt
`
`gcaaggtctc caacaaagge
`
`ctcecagcce
`
`ecatcgagaa
`
`aaccatctcc
`
`1080
`
`aaaaccaaag ggcagcccecg agaaccacag gtgtacaccc
`
`tgeccccatc
`
`cegggaggag
`
`1140
`
`atgaccaaga accaggtcag cctgacctge ctggtcaaag gcttctacce
`
`cagegacatc
`
`1200
`
`gcegtggagt gggagagcaa tgggcagecg gagaacaact acaagaccac
`
`acctcccatg
`
`1260
`
`ctggactcceg acggctcctt cttcctctac agcaagctca ccgtgqgacaa
`
`gagcaggtgg
`
`1320
`
`cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa
`
`ecactacacg
`
`1380
`
`Cagaagagec tctccctgte tecgqggtaaa tga
`
`1413
`
`Page 13 of 73
`
`Page 13 of 73
`
`

`

`U.S. Patent
`
`Nov.8, 2016
`
`Sheet 12 of 12
`
`US 9,487,581 B2
`
`FIG. 11B
`
`Ticilimumab (11.2.1) Heavy Chain Protein (SEQ ID NO:2)
`
`[QVOLVESGGG VVQPGRSLRL SCAASGFTFS SYGMHWVROA PGKGLEWVAV IWYDGSNKYY
`
`ADSVKGRFTI
`
`SRDNSKNTLY LOMNSLRAED TAVYYCARDP RGATLYYYYY GMDVWGQGTT
`
`VIVSSJASTKG PSVFPLAPCS RSTSESTAAL GCLVKDYFPE PVIVSWNSGA LTSGVHTFPA
`
`VLOSSGLYSL
`
`SSVVIVPSSN FGTOTYTCNVY DHKPSNTKVD KTVERKCCVE CPPCPAPPVA
`
`GPSVFLFPPK PKDTLMISRT PEVICVVVDV SHEDPEVOFN WYVDGVEVHN AKTKPREEQF
`
`NSTFRVVSVL
`
`TVVHQDWLNG KEYKCKVSNK GLPAPIEKTI SKIKGQPREP QVYTLPPSRE
`
`EMTKNQOVSLT
`
`CLVKGFYPSD IAVEWESNGQ PENNYKTTPP MLDSDGSFFL YSKLTVDKSR
`
`WQQGNVFSCS
`
`VMHEALHNHY TOKSLSLSPG K
`
`60
`
`120
`
`180
`
`240
`
`300
`
`360
`
`420
`
`451
`
`The variable region (SEQ ID NO:5)is depicted [between brackets] and the
`CDRsare underlined. CDR1 is indicated by SEQ ID NO:7, CDR2 by SEQ ID
`NO:8, and CDR3 by SEQ ID NO:9.
`
`Page 14 of 73
`
`Page 14 of 73
`
`

`

`US 9,487,581 B2
`
`1
`ANTI-CTLA-4 ANTIBODY COMPOSITIONS
`
`CROSS-REFERENCE TO RELATED PATENTS
`AND PATENT APPLICATIONS
`
`2
`physical stability relative to CTLA-4 antibody formulations
`previously disclosed in the literature.
`
`SUMMARY
`
`This application claims the benefit of U.S. Provisional
`Patent Application Ser. No. 60/659,766 filed Mar. 8, 2005;
`U.S. Provisional Patent Application Ser. No. 60/728,165
`filed Oct. 19, 2005; U.S. Provisional Patent Application Ser.
`No. 60/752,712 filed Dec. 20, 2005; and U.S. Provisional
`Patent Application Ser. No. 60/762,456 filed Jan. 26, 2006,
`all of which are incorporated by reference herein in their
`entireties.
`
`REFERENCE TO SEQUENCELISTING
`
`A revised sequencelisting in txt format is being submit-
`ted electronically via EFS-Web. The -txt file contains a
`sequenceentitled “PC33042A_SequenceListing.txt” created
`on Aug. 22, 2011 and having a size of 17 KB. The sequence
`listing contained in this .txt file is part of the specification
`and is herein incorporated by reference in its entirety.
`
`BACKGROUND OF THE INVENTION
`
`Cytotoxic T lymphocyte antigen-4 (“CTLA-4”) is a mem-
`ber of the immunoglobulin (“Ig”) superfamily of proteins.
`CTLA-4 acts to down regulate T-cell activation and maintain
`immunologic homeostasis. Blockade of CTLA-4 (e.g., by
`use of CTLA-4 antibodies) has been shown in animal
`models to improve the effectiveness of cancer immuno-
`therapy.
`Antibodies that bind to and inhibit the activity of CTLA-4
`have been reported in the literature. For example, U.S. Pat.
`No. 6,682,736 assigned to Pfizer, Inc. and Abgenix, Inc.,
`reports several human monoclonal antibodies to CTLA-4,
`including a CTLA-4 antibody having the heavy and light
`chain amino acid sequences of antibody 11.2.1, now known
`as Ticilimumab™, A hybridomacell line producing anti-
`body 11.2.1 was deposited under ATCC Accession No.
`PTA-5169. U.S. Pat. No. 5,977,318 assigned to Bristol-
`Myers Squibb Company, reports another monoclonalanti-
`body, which recognizes and binds the extracellular domain
`of CTLA-4, thereby preventing the binding of CTLA-4 to
`he B7
`antigen. U.S.
`Published Application No.
`20050201994 assigned to Medarex,
`Inc.
`reports several
`human sequence antibodies to CTLA-4, including one now
`referred to as Ipilimumab™.
`One possible mode of administering such CTLA-4 anti-
`bodies is by parenteral administration. For example, U.S.
`Pat. No. 6,682,736 reports an anti-CTLA-4 antibodyintra-
`venous formulationthatis a sterile liquid solution containing
`anti-CTLA-4 antibodies, 20 mMsodium acetate, 0.2 mg/ml
`polysorbate 80, and 140 mM sodium chloride at pH 5.5.
`Like other protein formulations, CTLA-4 antibody for-
`mulations are subject to the same concerns regarding chemi-
`cal and physical degradation of the antibody in the formu-
`ation over time. In general, CTLA-4 antibody formulations
`should exhibit acceptable chemical and physical stability
`under the expected range of storage and use conditions, ie.,
`the CTLA-4 antibody formulation should have a sufficient
`shelf life yet remain biologically active. Given the time and
`resources necessaryto produce a CTLA-4 antibody product,
`formulations that reduce productloss are desirable. Accord-
`ingly, the present application discloses novel CTLA-4anti-
`body formulations that exhibit improved chemical and/or
`
`
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`In one aspect, the present invention provides a liquid
`pharmaceutical composition comprising at least one anti-
`body comprising an amino acid sequencethatis at least 95%
`identical to a heavy chain amino acid sequence shown in
`SEQ ID NO: 2, and further comprising an amino acid
`sequencethatis at least 95% identical to a light chain amino
`acid sequence shown in SEQ ID NO: 4, wherein the anti-
`body binds to human CTLA-4; and a chelating agent.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the antibody is an IgG2
`antibody.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the antibody is a human
`antibody.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identical to a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the antibody comprises a V,,
`amino acid sequencethat utilizes a human V,,3-33 germline
`gene,
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the antibodyhas a V,, amino
`acid sequence that comprises human FR1, FR2, and FR3
`sequences that utilize a human V,, 3-33 gene family oper-
`ably linked in frame with a CDR1, a CDR2, and a CDR3
`sequence.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the antibody is an isolated
`antibody.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`
`Page 15 of 73
`
`Page 15 of 73
`
`

`

`US 9,487,581 B2
`
`
`
`3
`4
`and a chelating agent, wherein the antibody comprises a
`identical to a light chain amino acid sequence shown in SEQ
`heavy chain amino acid sequence comprising SEQ ID NO:
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`2 and a light chain amino acid sequence comprising SEQ ID
`and a chelating agent, wherein the antibodyis a recombinant
`NO: 4.
`antibody.
`The present invention also provides a composition com-
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`comprising an amino acid sequence that is at least 90%
`identical to a light chain amino acid sequence shown in SEQ
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the C-terminal lysine of the
`and a chelating agent, wherein the antibody specifically
`heavy chain of the antibody is not present.
`binds to a conformational epitope on the human CTLA-4
`The present invention also provides a composition com-
`polypeptide.
`prising at
`least one antibody comprising an amino acid
`The present invention also provides a composition com-
`sequence that is at least 90%identical to a heavy chain
`prising at
`least one antibody comprising an amino acid
`amino acid sequence shown in SEQ ID NO: 2, and further
`sequence that is at least 90% identical to a heavy chain
`comprising an amino acid sequence that is at least 90%
`amino acid sequence shown in SEQ ID NO:2, and further
`identicalto a light chain amino acid sequence shown in SEQ
`comprising an amino acid sequence that is at least 90%
`ID NO:4, wherein the antibody binds to human CTLA-4;
`identicalto a light chain amino acid sequence shown in SEQ
`and a chelating agent, wherein the antibody comprises a
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`monoclonal IgG2 anti-CTLA-4 antibody having the heavy
`and a chelating agent, wherein the antibody comprises a
`heavy chain amino acid sequence with at
`least 95%
`and light chain amino acid sequences of antibody 11.2.1.
`
`sequenceidentity to SEQ ID NO: 2 andalight chain amino The present invention also provides a composition com-
`25
`acid sequence with at least 95% sequence identity to SEQ ID
`prising at
`least one antibody comprising an amino acid
`NO: 4,
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`The present invention also provides a composition com-
`comprising an amino acid sequence that is at least 90%
`prising at
`least one antibody comprising an amino acid
`identical to a light chain amino acid sequence shown in SEQ
`sequence that is at least 90% identical to a heavy chain
`ID NO:4, wherein the antibody binds to human CTLA-4;
`amino acid sequence shown in SEQ ID NO:2, and further
`and a chelating agent, wherein the antibody has the same
`comprising an amino acid sequence that is at least 90%
`heavy and light chain amino acid sequencesas the antibody
`identicalto a light chain amino acid sequence shown in SEQ
`produced by hybridomacell line 11.2.1.4 deposited under
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`ATCC Accession No. PTA-5169.
`and a chelating agent, wherein the antibody comprises a
`heavy chain amino acid sequence with at
`least 99%
`The present invention also provides a composition com-
`
`
`sequence identity to SEQ ID NO: 2 andalight chain amino prising at least one antibody comprising an amino acid
`acid sequencewith at least 99% sequenceidentity to SEQ ID
`sequence that is at least 90% identical to a heavy chain
`NO: 4,
`amino acid sequence shown in SEQ ID NO: 2, and further
`The present invention also provides a composition com-
`comprising an amino acid sequence that is at least 90%
`prising at
`least one antibody comprising an amino acid
`identicalto a light chain amino acid sequence shown in SEQ
`sequence that is at least 90% identical to a heavy chain
`ID NO:4, wherein the antibody binds to human CTLA-4;
`amino acid sequence shown in SEQ ID NO:2, and further
`and a chelating agent, wherein the antibodyis ticilimumab.
`comprising an amino acid sequence that is at least 90%
`The present invention also provides a composition com-
`identicalto a light chain amino acid sequence shown in SEQ
`prising at
`least one antibody comprising an amino acid
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`sequence that is at least 90%identical to a heavy chain
`and a chelating agent, wherein the antibody comprises a
`amino acid sequence shown in SEQ ID NO: 2, and further
`heavy chain amino acid sequence that comprises the variable
`comprising an amino acid sequence that is at least 90%
`
`region of SEQ ID NO: 2 andalight chain amino acid identicalto a light chain amino acid sequence shown in SEQ
`sequence that comprises the variable region SEQ ID NO: 4.
`ID NO:4, wherein the antibody binds to human CTLA-4;
`The present invention also provides a composition com-
`and a chelating agent, wherein the chelating agentis selected
`prising at
`least one antibody comprising an amino acid
`from the group consisting of aminopolycarboxylic acids,
`sequence that is at least 90%identical to a heavy chain
`hydroxyaminocarboxylic acids, EDTAsalts and derivatives,
`amino acid sequence shown in SEQ ID NO:2, and further
`N-substituted glycines, deferoxamine derivatives and mix-
`tures thereof.
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`The present invention also provides a composition com-
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`prising at
`least one antibody comprising an amino acid
`and a chelating agent, wherein the antibody comprises a
`sequence that is at least 90% identical to a heavy chain
`heavy chain variable region amino acid sequence that com-
`amino acid sequence shown in SEQ ID NO: 2, and further
`prises SEQ ID NO: 5 anda light chain variable region amino
`comprising an amino acid sequence that is at least 90%
`acid sequence that comprises the comprises SEQ ID NO: 6.
`identicalto a light chain amino acid sequence shown in SEQ
`The present invention also provides a composition com-
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`prising at
`least one antibody comprising an amino acid
`and a chelating agent, wherein the chelating agent is selected
`sequence that is at least 90% identical to a heavy chain
`from the group consisting of ethylenediaminetetraacetic
`amino acid sequence shown in SEQ ID NO:2, and further
`acid, diethylenetriamine pentaacetic acid 5, nitrilotriacetic
`comprising an amino acid sequence that is at least 90%
`acid, N-2-acetamido-2-iminodiacetic acid, bis(aminoethyl)
`identicalto a light chain amino acid sequence shown in SEQ
`glycolether, N,N,N',N'-tetraacetic acid, trans-diaminocyclo-
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`hexane tetraacetic acid, glutamic acid, aspartic acid, N-hy-
`
`40
`
`10
`
`15
`
`20
`
`30
`
`35
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Page 16 of 73
`
`Page 16 of 73
`
`

`

`US 9,487,581 B2
`
`5
`
`
`
`N,N-bis-
`acid,
`droxyethyliminodiacetic
`hydroxyethylglycine, N-(trishydroxymethylmethyl)
`10
`glycine, glycylglycine, 2-(2-amino-2-oxoethyl) aminoeth-
`ane sulfonic acid, deferoxamine, deferoxamine mesylate,
`dipotassium edetate, disodium edetate, edetate calcium diso-
`dium, sodiumedetate, trisodium edetate, potassium edetate,
`citric acid, sodium citrate, anhydrouscitric acid, trisodium-
`citrate-dihydrate, niacinamide, sodium desoxycholate, and
`mixtures thereof.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the chelating agent is EDTA.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical
`to a heavy chain
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, further comprising a buffer.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical
`to a heavy chain
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`identical to a light chain amino acid sequence shown in SEQ
`D NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, further comprising a buffer, wherein
`he buffer is selected from the group consisting of acetate,
`succinate, gluconate, citrate, histidine, acetic acid, phos-
`phate, phosphoric acid, ascorbate, tartartic acid, maleic acid,
`glycine, lactate, lactic acid, ascorbic acid, imidazole, bicar-
`bonate and carbonic acid, succinic acid, sodium benzoate,
`benzoic acid, gluconate, edetate, acetate, malate, imidazole,
`ris, phosphate, and mixtures thereof.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, further comprising a buffer, wherein
`the buffer comprises histidine.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, further comprising histidine, wherein
`the histidine comprises L-histidine or D-histidine.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO:2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO: 4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, further comprising histidine, wherein
`the histidine comprises L-histidine.
`
`
`
`6
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90%identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identical to a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the composition contains a
`concentration of antibodies ranging from about 0.1 to about
`200 mg/ml.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the composition contains a
`concentration of antibodies of about 20 mg/ml.
`The present invention also provides a composition com-
`prising at
`least one antibody comprising an amino acid
`sequence that is at least 90% identical to a heavy chain
`amino acid sequence shown in SEQ ID NO: 2, and further
`comprising an amino acid sequence that is at least 90%
`identicalto a light chain amino acid sequence shown in SEQ
`ID NO:4, wherein the antibody binds to human CTLA-4;
`and a chelating agent, wherein the composition further
`comprises at least one excipient selected from the