`
`(54) HER2 ANTIBODY COMPOSITION
`.
`_
`juneshins Kao, San Mateo, CA
`(US); Martin Vanderlaan, San
`Francisco, CA (US)
`
`Inventors:
`
`(75)
`
`(73) Assignee: Genentech, Inc., South San Francisco,
`CA (US)
`.
`.
`.
`.
`Subject to any disclaimer, the termofthis
`patent is extended or adjusted under 35
`U.S.C. 154(b) by0 days.
`
`.
`Notice:
`
`«
`ie
`
`(21)
`
`12) United States Patent
`Kaoetal.
`
`10) Patent No:
`(45) Date of Patent:
`
`US 7,560,111 B2
`9
`9
`Jul. 14, 2009
`
`US007560111B2
`
`7,371,379 B2
`2001/0014326 AL
`2002/0001587 AL
`2003/0147884 Al
`2003/0170234 AL
`
`2003/0202972 Al
`2004/0037823 A9
`2004/0037824 Al
`2004/0106161 Al
`D004/0258685
`AL
`20040258085
`AL
`z
`2005/0002928 AL
`
`5/2008 Baughman etal.
`(2001 Andyaetal.
`1/2002 Ericksonet al.
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`6/2004 Bossenmaier et al.
`12/2004 B
`1
`Lid
`runetta et al.
`i
`ellmann
`1/2005 Hell
`
`2005/0208043 Al
`2005/0238640 Al
`2005/0244417 AL
`2006/0013819 AL*
`2006/0034840 AL
`2006/0034842 Al
`2006/0073 143 Al
`2006/0083739 Al
`2006/0088523 Al
`2006/0121044 Al
`
`9/2005 Adams otal.
`10/2005 Sliwkowski
`11/2005 Ashkenazi et al.
`1/2006 Kelsey oc 424/155.1
`/2006 Agus
`2/2006 Adamsetal.
`4/2006 Adamset al.
`4/2006 Sliwkowski
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`6/2006 Amleret al.
`
`Appl. No.: 11/182,908
`.
`Filed:
`
`(22)
`(65
`
`Jul. 15, 2005
`Prior Publication Data
`US 2006/0018899 Al
`Jan. 26, 2006
`Related U.S. Application Data
`.
`.
`.
`(60) sesional application No. 60/590,202,filed on Jul.
`,
`.
`
`(51)
`
`Int. Cl.
`(2006.01)
`AGIK 39395
`(2006.01)
`CO7K 16/00
`(2006.01)
`CO7K 1630
`(52) US.Ch oe. 424/138.1; 424/143.1; 530/388.8;
`530/388.85; 530/389.7
`(58) Field of Classification Search ............... 530/387.7
`See application file for complete searchhistory.
`.
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`FOREIGN PATENT DOCUMENTS
`
`EP
`
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`
`5/2003
`
`(Continued)
`OTHER PUBLICATIONS
`
`Aguset al., “Clinical Activity in a Phase I Trial of HER2-Targeted.
`rhuMAb 2C4 (pertuzumab) in Patients with Advanced. Solid Malig-
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`
`(Continued)
`
`Primary Examiner—Laura B Goddard
`(74) Atiorney, Agent, or Firm—Wendy M.Lee
`
`(57)
`
`ABSTRACT
`
`A composition comprising a main species HER2 antibody
`that binds to domain TI of HER2, and an amino acid sequence
`variant thereof comprising an amino-terminal leader exten-
`sion is disclosed. Pharmaccutical formulations comprising
`the composition, and therapeutic uses for the compositionare
`alsa disclosed.
`
`12 Claims, 23 Drawing Sheets
`
`Pfizer v. Genentech
`IPR2017-01488
`Genentech Exhibit 2043
`
`Pfizer v. Genentech
`IPR2017-01488
`Genentech Exhibit 2043
`
`

`

`US 7,560,111 B2
`
`Page 2
`
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`Stancovski et al., “Mechanistic Aspects of the Opposing Effects of
`Monoclonal Antibodies to the ERBB2 Receptor on Tumor Growth”
`Proc. Natl. Acad. Sci. USA 88(19):869 1-8695 (Oct. 1, 1991).
`Tagliabueetal., “Selection of Monoclonal Antibodies Which Induce
`Internalization and Phosphorylation of p185”“*? and Growth Inhi-
`bition of Cells With HER2/NEU Gene Amplification” International
`Journal ofCancer 47(6):933-937 (Apr. 1, 1991).
`
`Vitetta and Uhr, “Monoclonal Antibodies as Agonists: An Expanded
`Role for Their Use in Cancer Therapy” Cancer Research
`54(20):5301-5309 (Oct. 15, 1994).
`Weiner et al., “Expression of the neu Gene-encoded Protein
`(P185"°) in Human Non-Small Cell Carcinomasof the Lung” Can-
`cer Research 50(2):42 1-425 (Jan. 15, 1990).
`Williams et al., “Expression of c-erbB-2 in Human Pancreatic
`Adenocarcinomas” Pathobiology 59(1):46-52 (1991).
`Wuet al., “Apoptosis Induced By an Anti-Epidermal Growth Factor
`Receptor Monoclonal Antibody in a Human Colorectal Carcinoma
`Cell Line and Its Delay By Insulin” Journal ofClinical Investigation
`95(4): 1897-1905 (Apr. 1995).
`Xuetal., “Antibody-Induced Growth Inhibition is Mediated Through
`Immunochemically and Functionally Distinct Epitopes on the
`Extracellular Domain of the c-erbB-2 (HER-2/neu) Gene Product
`p185” International Journal ofCancer 53(3):401-408 (Feb.1, 1993).
`Yokota et al., “Amplification of c-erbB-2 Oncogene in Human
`Adenocarcinomasin Vivo” Lancet 1(8484):765-767 (Apr. 5, 1986).
`Yonemuraetal., “Evaluation of Immunoreactivity for erbB-2 Protein
`as a Marker of Poor Short Term Prognosis in Gastric Cancer” Cancer
`Research 51(3): 1034-1038 (Feb. 1, 1991).
`Zhanget al., “Neuregulin-3 (NRG3): A novel neural tissue-enriched
`protein that binds and activates ErbB4”Proc. Natl. Acad. Sci. USA
`94:9562-9567 (Sep. 22, 1997).
`Zhau et al., “Amplification and Expression of the cOerb B-2/neu
`Proto-Oncogene
`in Human Bladder Cancer” Molecular
`Carcinogenesis 3(5):254-257 (1990).
`
`* cited by examiner
`
`

`

`U.S. Patent
`
`
`
`AXSOADOICGO1TASISWNLdTALTEINDOAAODOOATHYINGIH.LAdSVdTHTINALDLOAOL
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`Jul. 14, 2009
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`Sheet 1 of 23
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`US 7,560,111 B2
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`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 2 of 23
`
`US 7,560,111 B2
`
`VARIABLE LIGHT
`
`2C4
`
`574
`
`40
`30
`20
`10
`DIVMTOSHKIMSTSVGDRVSITC [KASQDVSIGVA] WYQORP
`kk
`kkKkek
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`DIQMTOSPSSLSASVGDRVTITC [KASQDVSIGVA] WYQOKP
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`DIOMTOSPSSLSASVGDRVTITC [RASQSISNYLA] WYQOKP
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`574
`
`80
`70
`60
`90
`GOSPKLLIY [SASYRYT] GVPDRFTGSGSGTDFTFTISSVOA
`xk
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`* k
`
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`k KKK
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`GKAPKLLIY [AASSLES] GVPSRFSGSGSGTDFTLTISSLOP
`
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`90
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`kok
`k
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`kaK OX
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`hum «I
`
`EDFATYYC [QQYNSLPWT] FGQGTKVEIK (SEQ ID NO:5)
`
`FIG._2A
`
`VARIABLE HEAVY
`
`2C4
`
`574
`
`40
`30
`20
`10
`EVOLQOSGPELVKPGTSVKISCKAS [GFTFTDYTMD] WVKQOS
`kk
`Kk
`*
`k kkK
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`kk kek
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`2C4
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`574
`
`80
`70
`60
`a
`50
`HGKSLEWIG [DVNPNSGGSIYNORFKG] KASLTVDRSSRIVYM
`x ak
`kkk ok
`khkk ok
`
`PGKGLEWVA [DVNPNSGGSIYNORFKG}] RFTLSVDRSKNTLYL
`KKK KKK KkEK
`kok
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`hum III
`
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`
`2C4
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`574
`
`110
`100ab
`90
`abc
`ELRSLTFEDTAVYYCAR [NLGPSFYFDY] WGOGTTLTVSS (SEQ ID NO:2)
`kik
`kk
`kk
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`QMNSLRAEDTAVYYCAR [NLGPSFYFDY] WGQGTLVTVSS
`KaEKKKKEKE
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`(SEQ ID NO:4)
`
`hum III
`
`QMNSLRAEDTAVYYCAR [GRVGYSLYDY] WGQGTLVTVSS
`
`(SEQ ID NO:6)
`
`FIG._2B
`
`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 3 of 23
`
`US 7,560,111 B2
`
`Amino Acid Sequencefor PertuzumabLight Chain
`
`10
`1
`|
`|
`|
`|
`DIQMTOSPSSLSASVGDRVTITCKASQDVSIGVAWYQOKPGKAPKLLIYSAS¥RYTGVPS
`
`30
`
`40
`
`50
`
`60
`
`20
`
`70
`
`80
`
`90
`
`100
`
`110
`
`120
`
`|
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`
`ma
`
`net
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`| nt
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`| aa
`
`150
`
`mat
`
`190
`
`200
`
`210
`
`LSKADYEKHEVYAC|THOGLSSPUTKSFURGEC
`
`F,/G 3A
`
`Amino Acid Sequencefor Pertuzumab Heavy Chain
`Topo OR
`
`EVOLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVROQAPGKGLEWVADVNPNSGGSIY
`
`110
`90
`80
`70
`|
`|
`|
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`|
`|
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`NORFKGRFTLSVDRSKNTLYLOMNSLRAEDTAVYYCARNLGPSFYFDYWGQGTLVTVSSA
`
`100
`
`120
`
`130
`
`140
`
`150
`
`160
`
`170
`
`180
`
`oTKGbSVFPLAPSSSTSGYTAALGCLVKDYFPEMVTVSINSGALTSGVITFPAVLOSSE
`
`190
`
`200
`
`210
`
`220
`
`230
`
`240
`
`LysLdsvvrbpssslGroTyTONVIHKPSNTKVDLKVEPLScDKtHTCELCPAPELLGGD
`250
`260
`270
`280
`290
`300
`SVFLEPPKPLDTLMSRTPRVTCVVDVSHEDPEVKENAYVDGVEVENAKTKPREBOYNS
`
`310
`
`320
`
`330
`
`340
`
`350
`
`360
`
`TYRVVSVLITVLHODULNGKLYRCKVSNKALPABTAETISKAKGOLREPOUVYTLPLSREEM
`
`370
`
`380
`
`390
`
`400
`
`410
`
`420
`
`TKNOVSLTCLVKGFYPSDIAVEWESNGOPUNNYK}TPPVLDSDGSFELYSKLTVOKSRU
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`448
`440
`430
`|
`|
`|
`|
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`|
`QGNVF SCSVMHEALHNHYTOKSLSLSPG
`
`FIG._3B
`
`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 4 of 23
`
`US 7,560,111 B2
`
`SE€T
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`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 5 of 23
`
`US 7,560,111 B2
`
`Sv
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`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 6 of 23
`
`US 7,560,111 B2
`
`
`Unbound Complex
`
`EGFR
`
`EGFR-EGF
`
`FIG._5
`
`
`
`
`
`Ligand-activatedEGFRHeterodimerizeswithHER22C4BindsattheHeterodimericBindingSite
`
`
`
`
`
`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 7 of 23
`
`US 7,560,111 B2
`
`
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`shemuiedPIVPUidVINOU}01€/ZHSHJOBuljdnog
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`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 8 of 23
`
`US 7,560,111 B2
`
`Pertuzumab
`
`Trastuzumab
`Herceptin
`
`Omnitarg
`
`¢ Binds in IV near JM.
`
`* Bindsin II at dimerization interface
`
`¢ Protects against receptor shedding
`
`* Does not prevent receptor shedding
`
`¢ Moderately affects receptor down-
`modulation
`
`* Moderately affects receptor down-
`modulation
`
`¢ Slight effect on HER2’s role as a
`coreceptor
`
`¢ Major effect on HER2’s role as a
`coreceptor
`
`FIG._7
`
`

`

`
`
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`U.S. Patent
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`Jul. 14, 2009
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`Sheet 9 of 23
`
`US 7,560,111 B2
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`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 10 of 23
`
`US 7,560,111 B2
`
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`U.S. Patent
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`Jul. 14, 2009
`
`Sheet 11 of 23
`
`US 7,560,111 B2
`
`
`
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`
`
`
`
`
`
`
`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 12 of 23
`
`US 7,560,111 B2
`
`Cation Exchange ChromatographyAnalysis of Native Pertuzumab
`50
`
`40
`
`30
`
`20
`
`Absorbance
`at 280 nm
`(mAU)
`
`400 LScale
`
`10
`10
`
`FIG._9A
`
`18
`
`20
`
`22
`
`24
`
`Time (min)
`
`26
`
`28
`
`30
`
`Cation Exchange Chromatography Analysis of CPB-Digested Pertuzumab
`50
`
`40
`
`30
`
`20
`
`Absorbance
`at 280 nm
`(mAU)
`
`FIG._9B
`
`Time (min)
`
`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 13 of 23
`
`US 7,560,111 B2
`
`Size Exclusion Chromatographic Analysis of Pertuzumab
`
`Absorbance
`at 280 nm
`(mAU)
`
`400 L Scale Run1
`
`Lot S9802A
`
`Reference
`
`Material
`
`FIG..10
`
`10
`
`15
`
`Time (min)
`
`20
`
`25
`
`

`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 14 of 23
`
`US 7,560,111 B2
`
`CE-SDS-LIF Analysis of Reduced Pertuzumab
`
`Relative
`Intensity
`
`300
`
`250
`
`200
`
`150
`
`100
`
`50
`
`0
`
`FIG._11A
`
`250
`
`200
`
`150
`
`100
`
`50
`
`Detector
`Response
`
`400 L Scale Run 1
`
`FIG..11B
`
`7
`
`)
`11
`Migration Time (min)
`
`13
`
`15
`
`Non-glycosylated
`
`5
`
`6
`7
`8
`Migration Time (min)
`
`9
`
`10
`
`CE-SDS-LIF Analysis of Intact Pertuzumab
`
`Heavy Chain
`
`
`

`

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`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 15 of 23
`
`US 7,560,111 B2
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`09
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`U.S. Patent
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`Jul. 14, 2009
`
`Sheet 16 of 23
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`US 7,560,111 B2
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`

`U.S. Patent
`
`Jul. 14, 2009
`
`Sheet 17 of 23
`
`US 7,560,111 B2
`
`CE Analysis of N-linked Oligosaccharides Released from Pertuzumab
`
`33705., ChanA
`33703., ChanA
`33702., ChanA
`
`Mans
`GO-F
`
`400 L Scale
`Run1
`Lot $9802A
`
`Man6 + G1-1
`
`G1\
`
`1
`
`|
`
`G1(1-6)
`A G1(1-3) G2
`
`28
`
`29
`
`Time (min)
`
`30
`
`31
`
`32
`
`33
`
`3544
`
`30
`
`7 2
`
`0
`
`15
`
`10
`
`5
`
`0
`
`RFU
`
`FIG._ 13
`
`50000
`
`40000
`
`30000
`
`Relative
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`20000
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`Overlaid Traces: Aligned 40—)
`10000
`
`Reference Material
`
`25
`
`26
`
`27
`
`Positive Mode MALDI-TOF MassSpectra of Released
`Neutral Oligosaccharides Released from Pertuzumab
`
`400 L Scale Run 1
`
`Lot S9802A
`
`FIG._15
`
`M/Z
`
`1200
`
`1400
`
`1600
`
`1800
`
`2000
`
`2200
`
`

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`U.S. Patent
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`Jul. 14, 2009
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`Sheet 18 of 23
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`US 7,560,111 B2
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`U.S. Patent
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`Jul. 14, 2009
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`Sheet 19 of 23
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`US 7,560,111 B2
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`U.S. Patent
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`NIVHOLHS!
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`Jul. 14, 2009
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`SvO€|T
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`US 7,560,111 B2
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`U.S. Patent
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`Jul. 14, 2009
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`Sheet 21 of 23
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`US 7,560,111 B2
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