`
`(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.
`8/2003 Paton etal.
`9/2003 Hellmann
`
`10/2003 Andyaet al.
`/2004 Paton etal.
`2/2004 Baughmanetal
`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
`4/2006 Andyaetal.
`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.
`.
`References Cited
`U.S. PATENT DOCUMENTS
`
`(56)
`
`4,968,603 A
`5,183,884 A
`5,480,968 A
`5,641,869 A
`5,677,171 A
`$8,720,937 A
`5,720,954 A
`5,725,856 A
`5,770,195 A
`5,772,997 A
`5,783,186 A
`5,821,337 A
`5,824,311 A
`6,054,297 A
`6,165,464 A
`6,267,958 Bl
`6,339,142 Bl
`6,387,371 Bl
`6,399,063 Bl
`6,407,213 Bl
`6,627,196 Bl
`6,639,055 Bl
`6,685,940 B2
`6,719,971 Bl
`6,800,738 Bl
`6,821,515 Bl
`6,949,245 BL*
`7,041,292 Bl
`7,097,840 B2
`7,371,376 Bl
`
`11/1990 Slamonetal.
`2/1993 Kraus ctal.
`1/1996 Kraus ctal.
`6/1997 Vandlen et al
`10/1997 IIudziak etal.
`2/1998 IIudziak etal.
`2/1998 Hudziak etal.
`3/1998 Hudziak etal.
`6/1998 Hudziak etal.
`6/1998 Hudziak etal.
`7/1998 Arakawaet al,
`10/1998 Carteretal,
`10/1998 Greeneet al.
`4/2000 Carteret al,
`12/2000 Hudziak ctal.
`7/2001 Andyaetal.
`{2002 Baseyetal.
`§/2002 IIudziak etal.
`(2002 Hudziak et al.
`6/2002 Carteret al.
`9/2003 Baughman etal.
`10/2003 Carteretal.
`2/2004 Andyaetal.
`/2004 Carter et al.
`10/2004 Carteretal.
`11/2004 Clelandetal.
`9/2005 Sliwkowski.............. 424/143.1
`5/2006 Sliwkowski
`8/2006 Ericksonet al.
`5/2008 Fendly
`
`2006/0165702 Al
`2006/0188509 Al
`2006/0193854 Al
`2006/0198843 Al
`2006/0204505 Al
`
`7/2006 Allison etal.
`8/2006 Deryncketal.
`8/2006 Adamsetal.
`9/2006 Adamset al.
`9/2006 Sliwkowskietal.
`
`(Continued)
`
`
`
`FOREIGN PATENT DOCUMENTS
`
`EP
`
`1308455 A2
`
`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-
`nancies” (Slides presented at the 2003 ASCO Annual Meeting) pp.
`1-32 (2003).
`
`(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
`
`Celltrion v. Genentech
`IPR2017-01374
`Genentech Exhibit 2043
`
`Celltrion v. Genentech
`IPR2017-01374
`Genentech Exhibit 2043
`
`
`
`US 7,560,111 B2
`
`Page 2
`
`U.S. PATENT DOCUMENTS
`
`2006/0210561 Al
`2006/0216285 Al
`2006/0228745 Al
`2006/0275305 Al
`2006/0275306 Al
`2007/0009976 Al
`2007/0020261 Al
`2007/0026001 Al
`2007/0037228 Al
`2007/0077243 Al
`2007/0166753 Al
`2007/0184055 Al
`2007/0202516 Al
`2007/0224203 Al
`2007/0269429 Al
`2007/0292419 Al
`2008/0038271 Al
`2008/0050373 Al
`2008/0050385 Al
`2008/0102069 Al
`2008/0112957 Al
`2008/0112958 Al
`2008/0160026 Al
`
`9/2006 Baughman etal.
`9/2006 Adamset al.
`10/2006 Mass
`12/2006 Bryant
`12/2006 Andyaetal.
`1/2007 Lenz etal.
`1/2007 Sliwkowskiet al.
`2/2007 Ashkenaziet al.
`2/2007 Moecksetal.
`4/2007 Carteret al.
`7/2007 Mass
`8/2007 Sliwkowski
`8/2007 Mass
`9/2007 Friess et al.
`11/2007 Kelsey et al.
`12/2007 Hellmann
`2/2008 Amleretal.
`2/2008 Cohen
`2/2008 Friess et al.
`5/2008 Friesset al.
`5/2008 Fendly et al.
`5/2008 Mass
`7/2008 Ashkenaziet al.
`
`FOREIGN PATENT DOCUMENTS
`
`OTHER PUBLICATIONS
`
`Journal of Clinical Oncology (Abstract 4609; 41st Annual Meeting
`of ASCO)23(16S):405s (Jun. 1, 2005).
`Friesset al., “Combination treatment with erlotinib and pertuzumab
`against human tumorxenograftsis superior to monotherapy” Clinical
`Cancer Research 11(14):5300-5309 (Jul. 15, 2005).
`Friesset al., “In vivoactivity of recombinant humanized monoclonal
`antibody 2C4 in xenografts is independent of tumor type and degree
`ofHER2 overexpression” European Journal ofCancer (Abstract 496
`from the EORTC-NCI-AACR conference in Frankfurt, Germany
`Nov. 19-22, 2002.) 38(Suppl. 7):S 149 (2002).
`Gordon et al., “Clinical activity of pertuzumab (rhuMab 2C4) in
`advanced,refractory or recurrent ovarian cancer (OC), andthe role of
`HER2 activation status” Journal of Clinical Oncology (Abstract
`#5051 fromthe 41st Annual Meeting ofASCO) 23(16S):467s (Jun. 1,
`2005).
`Gordon et al., “Clinical activity of pertuzumab (rhuMab 2C4) in
`advanced, refractory or recurrent ovarian cancer and the role of
`HER2 activation status” (Poster #5051 from the 41st Annual Meeting
`of the American Society of Clinical Oncology (ASCO)) (May 15,
`2005).
`Hasmann et al., “Pertuzumab (Omnitarg) Potentiates Antitumor
`Effects on NSCLS Xenografts without Increasing Toxicity when
`Combined with Cytotoxic Chemotherapeutic Agents” American
`Association for Cancer Research (Abstract #B213; supplement to
`Clinical Cancer Research) 9(16) (Dec. 1, 2003).
`Nahta et al., “The HER-2-targeting antibodies trastuzumab and
`pertuzumab synergistically inhibit the survival ofbreast cancer cells”
`1/1994
`WO 94/00 136
`WO
`Cancer Research 64(7):2343-2346 (Apr. 1, 2004).
`10/1994
`WO 94/22478
`WO
`Spiridonet al., “Targeting multiple Her-2 epitopes with monoclonal
`4/1998
`WO 98/17797 Al
`WO
`antibodies results in improved antigrowth activity of a human breast
`cancer cell line in vitro and in vivo” Clinical Cancer Research
`
`WO WO 00/69460 Al—11/2000
`WO
`WO 01/00238 Al
`1/2001
`8(6): 1720-1730 (Jun. 2002).
`
`WO WO 03/087131 A2—10/2003
`Vajdos et al., “Comprehensive functional mapsof the antigen-bind-
`ing site of an anti-ErbB2 antibody obtained with shotgun scanning
`mutagenesis” Journal ofMolecular Biology 320(2):415-428 (Jul. 5,
`2002).
`Aasland et al., “Expression of Oncogenes in Thyroid Tumours:
`Coexpression of c-erbB2/neu and c-erbB”British Journal ofCancer
`57(4):358-363 (Apr. 1988).
`Aguset al., “Clinical Activity in a Phase I Trial of HER-2-Targeted.
`thuMAb2C4 (pertuzumab)in Patients with Advanced Solid Malig-
`nancies (AST)”Proceedings oftheAmerican Associationfor Cancer
`Research (Abstract No. 771) 22:192 (2003).
`Agus et al., “Targeting ligand-activated ErbB2 signaling inhibits
`breast and prostate tumor growth” Cancer Cell 2(2):127-137 (Aug.
`2002).
`Arteaga etal., “p185°°??? Signaling Enhances Cisplatin-induced
`Cytotoxicity in Human Breast Carcinoma Cells: Association
`Between an Oncogenic Receptor Tyrosine Kinase and Drug-induced.
`DNARepair” Cancer Research 54(14):3758-3765 (Jul. 15, 1994).
`Bacuset al., “Differentiation of Cultured Human Breast Cancer Cells
`(AU-565 and MCF-7) Associated With Loss of Cell Surface HER-2/
`neu Antigen” Molecular Carcinogenesis 3(6):350-362 (1990).
`Bacusetal., “Tumor-inhibitory Monoclonal Antibodies to the HER-
`2/Neu Receptor Induce Differentiation of Human Breast Cancer
`Cells” Cancer Research 52(9):2580-2589 (May 1, 1992).
`Baselga and Mendelsohn, “Receptor Blockade With Monoclonal
`Antibodies As Anti-Cancer Therapy” Pharmac. Ther. 64:127-154
`(1994).
`Baselga etal., “Phase II Study of Weekly Intravenous Recombinant
`Humanized Anti-p185”“*? Monoclonal Antibody in Patients With
`HER2/neu-Overexpressing Metastatic Breast Cancer” J Clin.
`Oncol. 14(3):737-744 (Mar. 1996).
`Borst et al., “Oncogene Alterations in Endometrial Carcinoma”
`Gynecologic Oncology 38(3):364-366 (Sep. 1990).
`Carraway and Cantley, “A Neu Acquaintance for ErbB3 and ErbB4:
`A Role for Receptor Heterodimerization in Growth Signaling” Cell
`78:5-8 (Jul. 15, 1994).
`Carraway et al., “Neuregulin-2, A New Ligand of ErbB3/ErbB4-
`Receptor Tyrosine Kinases” Nature 387:512-516 (May 1997).
`Chang etal., “Ligands For ErbB-Family Receptors Encoded By a
`Neuregulin-Like Gene” Nature 387:509-512 (May 29, 1997).
`
`Aguset al., “Efficacy andsafety of single agent pertuzumab (rhuMAb
`2C4) , a HER dimerization inhibitor, in hormonerefractory prostate
`cancer after failure of taxane-based therapy” Journal of Clinical
`Oncology (Abstract 4624 from the 41st Annual Meeting of ASCO)
`23(16S):408s (Jun. 1, 2005).
`Agus, D. et al., “Efficacy and safety of single agent pertuzumab
`(chuMAb2C4), a HER dimerization inhibitor, in hormonerefractory
`prostate cancer after failure of taxane-based therapy” (Poster 4624
`from the 41st Annual Meeting of the American Society of Clinical
`Oncology) (May 15, 2005).
`Amler et al., “Identification of a predictive expression pattern for
`phosphorylated HER2 as
`a potential diagnostic marker
`for
`pertuzumab (Omnitarg) activity in ovarian cancer” (Poster 4497 pre-
`sented. at the Apr. 2006 American Association for Cancer Research
`Meeting) (Apr. 2006).
`Bossenmaieret al., “Presence of HER2/HER3 heterodimers predicts
`antitumor effects of pertuzumab (Omnitarg) in different human
`xenograft models” Proc Am Assoc Cancer Res. (Abstract 5342)
`45:1232 (Mar. 2004).
`Corteset al., “Open label, randomized,phase II study of pertuzumab
`(Omnitarg) in patients with metastatic breast cancer (MBC)with low
`expression of HER2”(Poster 3068 from the 41st Annual Meeting of
`the American Society of Clinical Oncology (ASCO)) (May 15,
`2005).
`Corteset al., “Open label, randomized phase II study of pertuzumab
`(P) in patients (pts) with metastatic breast cancer (MBC) with low
`expression of HER2” Journal of Clinical Oncology (Abstract 3068
`from the 41st Annual Meeting ofASCO) 23(16s):208s (Jun. 1, 2005).
`de Bonoet al., “An open label, phase II, multicenter study to evaluate
`the efficacy and safety ofpertuzumab in chemotherapy-naive patients
`with Hormone-Refractory Prostate Cancer (HRPC)” (Poster 4609
`from the 41st Annual Meeting of the American Society of Clinical
`Oncology (ASCO)) (May 15, 2005).
`de Bonoet al., “An open label, phase II, multicenter, study to evaluate
`the efficacy and safety of pertuzumab (P) in chemotherapy naive
`patients (pts) with Hormone Refractory Prostate Cancer (HRPC)”
`
`
`
`US 7,560,111 B2
`
`Page 3
`
`Cohenet al., “Expression Pattern of the neu (NGL) Gene-Encoded
`Growth Factor Receptor Protein (p185"™) in Normal and. Trans-
`formed Epithelial Tissues of the Digestive Tract” Oncogene 4(1):81-
`88 (Jan. 1989).
`D’Souzaand Taylor-Papadimitriou., “Overexpression of ERBB2 in
`Human Mammary Epithelial Cells Signals Inhibition of Transcrip-
`tion of the E-Cadherin Gene” Proc. Natl. Acad. Sci. USA
`91(15):7202-7206 (Jul. 19, 1994).
`Drebin et al., “Down-Modulation of an Oncogene Protein Product
`and Reversion of the Transformed Phenotype by Monoclonal Anti-
`bodies” Cell 41(3):695-706 (Jul. 1985).
`Drebin et al., “Monoclonal Antibodies Reactive With Distinct
`Domains of the neu Oncogene-Encoded p185 Molecule Exert Syn-
`ergistic Anti-Tumor Effects In Vivo” Oncogene 2:273-277 (1988).
`Earp etal., “Heterodimerization and Functional Interaction Between
`EGF Receptor Family Members: A New Signaling Paradigm With
`Implications For Breast Cancer Research” Breast Cancer Res and
`Treatment 35:115-132 (1995).
`Fendly, B.M.et al., “Characterization of Murine Monoclonal Anti-
`bodies Reactive to Either the Human Epidermal Growth Factor
`Receptor or HER2/neu Gene Product” Cancer Research 50:1550-
`1558 (Mar. 1, 1990).
`Fukushige et al., “Localization of a Novel v-erbB-Related Gene,
`c-erbB2, on Human Chromosome 17 and Its Amplification in a
`Gastric Cancer Cell Line” Molecular & Cellular Biology 6(3):955-
`958 (Mar. 1986).
`Groenen et al., “Structure-Function Relationships for the EGF/
`TGF-a Family of Mitogens” Growth Factors 11:235-257 (1994).
`Guetal., “Overexpression of her-2/neu Human Prostate Cancer and
`Benign Hyperplasia” Cancer Letters 99:185-189 (1996).
`Guerin et al., “Overexpression of Either c-myc or c-erbB-2/neu
`Proto-Oncogenes in Human Breast Carcinomas: Correlation with
`Poor Prognosis” Oncogene Res. 3:21-31 (1988).
`Hancocket al., “A Monoclonal Antibody Against the c-erbB-2 Pro-
`tein Enhances the Cytotoxicity of cis-Diamminedichloroplatinum
`Against Human Breast and Ovarian Tumor Cell Lines” Cancer
`Research 51:4575-4580 (Sep. 1, 1991).
`Harari et al., “Neuregulin-4: A Novel Growth Factor That Acts
`Through the ErbB-4 Receptor Tyrosine Kinase” Oncogene 18:2681-
`2689 (1999).
`Harris, Reed, “The Ideal Chromatographic Antibody Characteriza-
`tion Method”(Slides 1-36, IBC Antibody Production Conference,
`Feb. 13, 2002).
`Harwerth et al., “Monoclonal Antibodies Against the Extracellular
`Domain ofthe erbB-2 Receptor Function as Partial Ligand Agonists”
`Journal of Biological Chemistry 267(21):15160-15167 (Jul. 25,
`1992).
`Holmeset al., “Identification of Heregulin, A Specific Activator of
`p1ss°?8*” Science 256:1205-1210 (May 22, 1992).
`Hudziak etal., “p185“*? Monoclonal Antibody Has Antiprolifera-
`tive Effects In Vitro and Sensitizes Human Breast Tumor Cells to
`Tumor Necrosis Factor” Molecular & Cellular Biology 9(3):1165-
`1172 (Mar. 1989).
`Kasprzyk et al., “Therapy of an Animal Model of Human Gastric
`Cancer Using a Combination of Anti-erbB-2 Monoclonal Antibod-
`ies” Cancer Research 52(10):2771-2776 (May 15, 1992).
`Kern etal., “p185”°“ Expression in Human Lung Adenocarcinomas
`Predicts Shortened Survival” Cancer Research 50(16):5184-5191
`(Aug. 15, 1990).
`King et al., “Amplification of a Novel v-erbB-Related Gene in a
`Human Mammary Carcinoma” Science 229:974-976 (Sep. 1985).
`Klapperet al., “A Subclass of Tumor-Inhibitory Monoclonal Anti-
`bodies to ErbB-2/HER2 Blocks Crosstalk With Growth Factor
`Receptors” Oncogene 14:2099-2109 (1997).
`Kotts et al., “Differential Growth Inhibition of Human Carcinoma
`Cells Exposed to Monoclonal Antibodies Directed against
`the
`Extracellular Domain of the HER2/ERBB2 Protooncogene”In Vitro
`(Abstract #176) 26(3):59A (1990).
`Krauset al., “Isolation and Characterization of ERBB3, A Third
`Memberof the ERBB/Epidermal Growth Factor Receptor Family:
`Evidence for Overexpression in a Subset of Human Mammary
`Tumors” Proc. Natl. Acad. Sci. USA 86:9193-9197 (Dec. 1989).
`
`
`
`Kumar etal., “Regulation of Phosphorylation of the c-erbB-2/HER2
`Gene Product by a Monoclonal Antibody and Serum Growth Fac-
`tor(s) in Human Mammary CarcinomaCells” Molecular & Cellular
`Biology 11(2):979-986 (Feb. 1991).
`Lee,“Transforming growth factor alpha: expression, regulation, and.
`biological activities” Pharm. Rev. 47(1):S1-85 (Mar. 1995).
`Lemke,G., “Neuregulins in Development” Molecular and Cellular
`Neurosciences 7:247-262 (1996).
`Levi et al., “The Influence of Heregulins on Human Schwann Cell
`Proliferation” J. Neuroscience 15(2):1329-1340 (Feb. 1995).
`Lewiset al., “Differential Responses of Human TumorCell Lines to
`Anti-p185““8?, Monoclonal Antibodies” Cancer
`Immunol.
`Immunother. 37:255-263 (1993).
`Lewis et al., “Growth Regulation of Human Breast and Ovarian
`TumorCells by Heregulin: Evidence for the Requirement of ErbB2
`as a Critical Component in Mediating Heregulin Responsiveness”
`Cancer Research 56:1457-1465 (Mar. 15, 1996).
`Maier et al., “Requirements for the Internalization of a Murine
`Monoclonal Antibody Directed against the Her-2/neu Gene Product
`c-erbB-2” Cancer Research 51(19):5361-5369 (Oct. 1, 1991).
`Masuietal., “Growth Inhibition of Human TumorCells in Athymic
`Mice by Anti-Epidermal Growth Factor Receptor Monoclonal Anti-
`bodies” Cancer Research 44(3): 1002-1007 (Mar. 1984).
`McCannet al., “c-erbB-2 Oncoprotein Expression in Primary Human
`Tumors” Cancer 65(1):88-92 (Jan. 1, 1990).
`McKenzie et al., “Generation and Characterization of Monoclonal
`Antibodies Specific for the Human neu Oncogene Product, p185”
`Oncogene 4:543-548 (1989).
`Morrissey et al., “Axon-Induced Mitogenesis of Human Schwann
`Cells Involves Heregulin and p185°”?*” Proc. Natl. Acad. Sci. USA
`92:1431-1435 (Feb. 1995).
`Myersetal., “Biological Effects of Monoclonal Antireceptor Anti-
`bodies Reactive with neu Oncogene Product, p185neu” Methods in
`Enzymology 198:277-290 (1991).
`Park et al., “Amplification, Overexpression, and Rearrangement of
`the erbB-2 Protooncogene in Primary Human Stomach Carcinomas”
`Cancer Research 49(23):6605-6609 (Dec. 1, 1989).
`Pietraset al., “Antibody to HER-2/neu Receptor Blocks DNA Repair
`After Cisplatin in Human Breast and Ovarian Cancer Cells”
`Oncogene 9:1829-1838 (1994).
`Plowman et al., “Heregulin Induces Tyrosine Phosphorylation of
`HER4/p180°?*” Nature (Letters to Nature) 366:473-475 (Dec. 2,
`1993).
`Plowman et al., “Ligand-Specific Activation of HER4/p180°"™", A
`Fourth Memberof the Epidermal Growth Factor Receptor Family”
`Proc. Natl. Acad. Sci. USA 90:1746-1750 (Mar. 1993).
`Porter, Jill, “The role of Analytical Comparability in the Global
`Approval of Zenapax” Case Studes—Biotech Manufacturing
`Changes (Slides 1-18, The 3rd International Conference: Strategic
`Use of Comparability Studies and Assays for Well Characterized
`Biologicals), Washington, D.C., Sep. 18-21, 2000.
`Reid etal., “Effects of Cell Culture Process Change on Humanized.
`Characteristics” (Poster presented at WCBP 2003 conference in San
`Francisco, Jan. 7-10, 2003, p. 1).
`Rosset al., “HER-2/neu Gene Amplification Status in Prostate Can-
`cer by Fluorescence in Situ Hybridization” Hum. Pathol. 28(7):827-
`833 (Jul. 1997).
`Rosset al., “Prognostic Significance of HER-2/neu Gene Amplifica-
`tion Status by Fluorescence In Situ Hybridization of Prostate Carci-
`noma” Cancer 79(11):2162-2170 (Jun. 1, 1997).
`Rouse et al., “Top Down Glycoprotein Characterization by High
`Resolution Mass
`Spectrometry
`and
`Its Application
`to
`Biopharmaceutical Develpoment” (Slides
`1-27, WCBP 2004
`meeting,Washington DC, Jan. 6-9, 2004).
`Sadasivan et al., “Overexpression of Her-2/Neu May Be An Indicator
`of Poor Prognosis in Prostate Cancer” J Urol. 150:126-131 (Jul.
`1993).
`Sarup et al., “Characterization of an Anti-P185’“*? Monoclonal
`Antibody that Stimulates Receptor Function and Inhibits TumorCell
`Growth” Growth Regulation 1:72-82 (1991).
`Schaeferet al., ““y-Heregulin: A Novel Heregulin Isoform That is an
`Autocrine Growth Factor for the Human Breast Cancer Cell Line,
`MDA-MB-175” Oncogene 15:1385-1394 (1997).
`
`
`
`US 7,560,111 B2
`
`Page 4
`
`Scott et al., “p185”“"" Signal Transduction in Breast Cancer Cells”
`Journal of Biological Chemistry 266(22):14300-14305 (Aug. 5,
`1991).
`Shawveretal., “Ligand-Like Effects Induced by Anti-c-erbB-2 Anti-
`bodies Do Not Correlate with and Are Not Required for Growth
`Inhibition of Human Carcinoma Cells” Cancer Research
`54(5):1367-1373 (Mar. 1, 1994).
`Shepard et al., “Monoclonal Antibody Therapy of Human Cancer:
`Taking the HER2 Protooncogene to the Clinic” J Clin. Immunol.
`11(3):117-127 (1991).
`Shields et al., “High Resolution Mapping of the Binding Site on
`Human IgGl for CyyRI, FeyRII, FeyRII, and FeRn and Design of
`IgGl Variants with Improved Binding to the FeyR*” JournalofBio-
`logical Chemistry 276(9):6591-6604 (2001).
`Slamon et al., “Human Breast Cancer: Correlation of Relapse and
`Survival with Amplification of the HER-2/neu Oncogene” Science
`235:177-182 (Jan. 9, 1987).
`Slamonetal., “Studies of the HER-2/neu Proto-Oncogene in Human
`Breast and Ovarian Cancer” Science 244:707-712 (May 12, 1989).
`Sliwkowski et al., “Coexpression of erbB2 and erbB3 Proteins
`Reconstitutes a High Affinity Receptor for Heregulin” Journal of
`Biological Chemistry 269(20):1466 1-14665 (May 20, 1994).
`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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`Sheet 1 of 23
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`
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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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`2C4
`
`574
`
`80
`70
`60
`90
`GOSPKLLIY [SASYRYT] GVPDRFTGSGSGTDFTFTISSVOA
`xk
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`* k
`
`GKAPKLLIY [SASYRYT] GVPSRFSGSGSGTDFTLTISSLOP
`k KKK
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`GKAPKLLIY [AASSLES] GVPSRFSGSGSGTDFTLTISSLOP
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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
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`k kkK
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`kk kek
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`EVQLVESGGGLVQPGGSLRLSCAAS [GFTFSSYAMS] WVROA
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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
`
`PGKGLEWVA {VISGDGGSTYYADSVKG] RFTISRDNSKNTLYL
`
`2C4
`
`574
`
`110
`100ab
`90
`abc
`ELRSLTFEDTAVYYCAR [NLGPSFYFDY] WGOGTTLTVSS (SEQ ID NO:2)
`kik
`kk
`kk
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`QMNSLRAEDTAVYYCAR [NLGPSFYFDY] WGQGTLVTVSS
`KaEKKKKEKE
`
`(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
`
`|
`RFSGSGSGTDFTLTISSLOPEDFATYYCQOYYIYPYTFGOGTKVEIKRTVAAPSVFIFPP
`
`SDEQLKSGTASVVCLLNNFY PREAKVOWKVDNALOSGNSQESVTEQDSKDSTYSLSSTLT
`
`ma
`
`net
`
`| nt
`
`| aa
`
`150
`
`mat
`
`190
`
`200
`
`210
`
`LSKADYEKHEVYAC|THOGLSSPUTKSFURGEC
`
`F,/G 3A
`
`Amino Acid Sequencefor Pertuzumab Heavy Chain
`Topo OR
`
`EVOLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVROQAPGKGLEWVADVNPNSGGSIY
`
`110
`90
`80
`70
`|
`|
`|
`|
`|
`|
`|
`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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`|
`|
`|
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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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`Jul. 14, 2009
`
`Sheet 5 of 23
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`US 7,560,111 B2
`
`Sv
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`
`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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`
`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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`
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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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`Jul. 14, 2009
`
`Sheet 11 of 23
`
`US 7,560,111 B2
`
`
`
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`
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`
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`
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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
`
`09
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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
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`Reference Material
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`Jul. 14, 2009
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`Sheet 18 of 23
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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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`NIVHOLHS!
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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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