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`MILTENYI BIOMEDICINE GmbH and MILTENYI BIOTEC INC.
`Petitioner
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`v.
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`THE TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA
`Patent Owner
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`
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`UNITED STATES PATENT AND TRADEMARK OFFICE
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`BEFORE THE PATENT TRIAL AND APPEAL BOARD
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`IPR Trial No. IPR2022 -
`U.S. Patent No. 9,464,140
`Issue Date: October 11, 2016
`
`Title: Compositions and Methods for Treatment of Cancer
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`DECLARATION OF DR. RICHARD P. JUNGHANS
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`IN SUPPORT OF PETITION FOR INTER PARTES REVIEW OF
`U.S. PATENT NO. 9,464,140
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`Miltenyi Ex. 1002 Page 1
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`TABLE OF CONTENTS
`Introduction ...................................................................................................... 1
`I.
`Education and Experience ............................................................................... 2
`II.
`III. Materials Considered ....................................................................................... 6
`IV. Technology Background .................................................................................. 6
`A.
`T Cells ................................................................................................... 7
`B.
`Chimeric Antigen Receptor T Cells (CAR T Cells) ........................... 10
`C.
`Engineering CAR T Cells ................................................................... 14
`D.
`Immunotherapy with CAR T Cells ..................................................... 18
`Person of Ordinary Skill in the Art (“POSA”) .............................................. 21
`V.
`VI. The ’140 Patent .............................................................................................. 22
`VII. Claim Construction ........................................................................................ 24
`A.
`Preamble – “treating cancer” ............................................................... 24
`B.
`“Anti-tumor effective amount” ........................................................... 25
`VIII. Prior Art ......................................................................................................... 28
`A.
`CAR T-Cell Therapies Known Prior to the Priority Date ................... 28
`1.
`The St. Jude CAR, Known as Anti-CD19-BB-ζ ...................... 28
`Sequences of the CAR Domains Were Known .................................. 34
`1.
`Anti-CD19 ScFv ....................................................................... 34
`2.
`Human CD8α Leader, Hinge, and Transmembrane
`Domains .................................................................................... 40
`4-1BB Costimulatory Domain .................................................. 46
`3.
`CD3-zeta Signaling Domain ..................................................... 46
`4.
`Animal and Clinical Studies of the Campana CAR ............................ 48
`1. Milone ....................................................................................... 48
`2.
`CART-19 ClinicalTrials.gov ..................................................... 51
`3.
`Porter ......................................................................................... 54
`D. A Pharmaceutical Composition of CAR T Cells ................................ 60
`1.
`Honsik ....................................................................................... 61
`IX. Legal Standards ............................................................................................. 62
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`C.
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`B.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`2.
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`B.
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`X. Ground 1: Independent Claim 1, and Dependent Claims 2, 6, 8-9, 11,
`16, 21-22, and 27-28 are rendered obvious by Campana in view of
`Nicholson, HONSIK, And CART-19 ClinicalTrials.gov .............................. 65
`A.
`Independent Claim 1 ........................................................................... 65
`1.
`Claim Limitations Directed to the Structure of the
`Claimed CAR T Cell ................................................................. 66
`Claim Limitations Directed to a Pharmaceutical
`Composition Comprising Administering an Anti-tumor
`Effective Amount of a Population of Human T cells ............... 74
`Dependent Claims ............................................................................... 93
`1.
`Claim 2: “anti-tumor effective amount of T cells is 104 to
`109 cells per kg body weight of a human in need of such
`cells” .......................................................................................... 93
`Claim 6: “wherein the transmembrane domain is CD8α
`transmembrane domain” ........................................................... 93
`Claims 8 and 9: “wherein the CAR further comprises a
`hinge domain” or “wherein the hinge domain is a CD8α
`hinge domain” ........................................................................... 94
`Claims 11 and 16: “wherein the 4-1BB costimulatory
`signaling region comprises the amino acid sequence of
`SEQ ID NO: 23” and “wherein the 4-1BB costimulatory
`signaling region comprises the nucleic acid sequence of
`SEQ ID NO: 17” ....................................................................... 94
`Claims 21 and 22: “wherein the T cells are T cells of a
`human having a cancer” or “wherein the cancer is a
`hematological cancer” ............................................................... 96
`Claims 27 and 28: “wherein the pharmaceutical
`composition further comprises a pharmaceutically
`acceptable carrier, diluent or excipient” and “wherein the
`pharmaceutical composition comprises a buffer” ..................... 98
`XI. Ground 2: Independent Claim 1, and Dependent Claims 2, 3, 6, 8-9,
`11, 13, 16, 21-22, and 27-28 are rendered obvious by Campana in
`view of Jensen, HONSIK, and CART-19 ClinicalTrials.gov ..................... 100
`A.
`Independent Claim 1 ......................................................................... 101
`1.
`Preamble of “[a] method of treating cancer in a human
`patient” .................................................................................... 101
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`2.
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`3.
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`4.
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`5.
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`6.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`2.
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`3.
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`4.
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`5.
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`6.
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`B.
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`“[b] administering to the human patient a pharmaceutical
`composition” ........................................................................... 101
`“[c] an anti-tumor effective amount of a population of
`human T cells” ........................................................................ 102
`“[d] wherein the T cells comprise a nucleic acid sequence
`encoding a chimeric antigen receptor (CAR)” ....................... 105
`“[e] wherein the CAR comprises a CD19 antigen binding
`domain comprising, from the amino to the carboxy
`terminus, a light chain variable region and a heavy chain
`variable region of SEQ ID NO:20” ......................................... 105
`“[f] wherein the CAR further comprises a transmembrane
`domain, a 4-1BB costimulatory signaling region, and a
`CD3 zeta signaling domain” ................................................... 109
`Dependent Claim 3: an “anti-tumor effective amount of T cells
`is 105 to 106 cells per kg body weight of a human in need of
`such cells” .......................................................................................... 109
`Claim 13: “wherein the CD19 antigen binding domain is
`encoded by a nucleic acid sequence comprising SEQ ID NO:
`14” ..................................................................................................... 113
`D. Dependent Claims 2, 6, 8-9, 11, 16, 21-22, and 27-28 ..................... 117
`XII. Ground 3: All Challenged Claims are Rendered Obvious by Campana
`in View of Milone, CART-19 ClinicalTrials.gov, Sequence Art
`(Jensen, Nicholson, Littman, Sadelain), AND Honsik ................................ 118
`A.
`Independent Claim 1 ......................................................................... 119
`1.
`Preamble of “[a] method of treating cancer” .......................... 119
`2.
`“[b] administering to the human patient a pharmaceutical
`composition” ........................................................................... 119
`“[c] anti-tumor effective amount of a population of
`human T cells” ........................................................................ 120
`“[d] wherein the T cells comprise a nucleic acid sequence
`encoding a chimeric antigen receptor (CAR)” ....................... 126
`“[e] wherein the CAR comprises a CD19 antigen binding
`domain comprising, from the amino to the carboxy
`terminus, a light chain variable region and a heavy chain
`variable region of SEQ ID NO:20” ......................................... 126
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`C.
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`3.
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`4.
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`5.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`6.
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`3.
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`4.
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`2.
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`C.
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`B.
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`C.
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`B.
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`“[f] wherein the CAR further comprises a transmembrane
`domain, a 4-1BB costimulatory signaling region, and a
`“CD3 zeta signaling domain” ................................................. 127
`Sequence Claims ............................................................................... 127
`1.
`The CD8α Hinge and Transmembrane of the ’140 Patent ..... 127
`2.
`Claims 12 and 17: “wherein the CD3 zeta signaling
`domain comprises the amino acid sequence of SEQ ID
`NO: 24” or “wherein the CD3 zeta signaling domain
`comprises the nucleic acid sequence of SEQ ID NO: 18”...... 138
`Claims 18 and 19: “wherein the CAR comprises amino
`acid sequence of SEQ ID NO: 12” or “wherein the CAR
`comprises nucleic acid sequence of SEQ ID NO: 8” ............. 140
`Statement About Nucleic and Amino Acid Sequences for
`CD8α and CD3-zeta Domains in Campana ............................ 142
`Vector/Promoter claims ..................................................................... 147
`1.
`Claims 23 and 24: “wherein the T cells comprise a vector
`that comprises the nucleic acid sequence” or “wherein
`the vector is a lentiviral vector” .............................................. 147
`Claims 25 and 26: “wherein the vector further comprises
`a promoter” or “wherein the promoter is an EF-1α
`promoter” ................................................................................ 148
`D. Dependent claims 2-3, 6, 8-9, 11, 13, 16, 21-22, 27- 28 ................... 150
`XIII. Ground 4: All Challenged Claims are Rendered Obvious by Campana,
`porter, Sequence Art, AND HONSIK ......................................................... 152
`A.
`Porter is Prior Art .............................................................................. 153
`1.
`Provisional Applications Do Not Provide Adequate
`Support for the Claims ............................................................ 153
`No Disclosure of SEQ ID NO: 20 in the Provisional
`Applications ............................................................................ 154
`Nicholson Teaches a VH-VL Order ....................................... 157
`3.
`All Challenged Claims are Rendered Obvious by Campana,
`Sequence Art, and Porter ................................................................... 164
`1.
`Independent Claim 1 ............................................................... 165
`Dependent Claims ............................................................................. 176
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`2.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`Dosing ..................................................................................... 176
`1.
`CAR T Cell Structure Claims ................................................. 178
`2.
`Sequence claims ...................................................................... 179
`3.
`Vector/Promoter Claims ......................................................... 183
`4.
`Pharmaceutical Composition Claims ...................................... 185
`5.
`XIV. Conclusion ................................................................................................... 187
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`I.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
`
`INTRODUCTION
`I, Richard Paul Junghans, PhD, MD, have been retained as an expert
`1.
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`witness on behalf of Miltenyi Biomedicine GmbH and Miltenyi Biotec Inc.
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`(collectively, “Petitioner”) in connection with its petition (“Petition”) for inter partes
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`review of U.S. Patent No. 9,464,140 (“the ’140 patent”) (Ex. 1001) (140 Patent).
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`The ’140 patent is owned by The Trustees of the University of Pennsylvania (“Patent
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`Owner”).
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`2.
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`I understand that this declaration is being submitted in support of
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`Petitioner’s petition for inter partes review of claims 1-19 and 21-28 (“Challenged
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`Claims”) of the ’140 patent, which was issued from U.S. Application No. 14/996,953
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`on October 11, 2016. The ’140 patent names Carl H. June, Bruce L. Levine, David
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`L. Porter, Michael D. Kalos, and Michael C. Milone as inventors.
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`3.
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`I have reviewed the ’140 patent and in my review, considered the
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`teachings of the scientific literature before December 9, 2011.
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`4.
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`I am being compensated at my usual and customary rate of $ 1,500 per
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`hour. My compensation is in no way dependent on the outcome of this case.
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`5.
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`In the last four years, I served as an expert in, and was deposed and/or
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`testified in connection with, the cases listed in Appendix B.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`II. EDUCATION AND EXPERIENCE
`A copy of my Curriculum Vitae, which describes my qualifications in
`6.
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`greater detail and includes a list of publications that I authored in the past 47 years,
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`is attached as Appendix A.
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`7.
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`I am an expert in the field of CAR T-cell therapy. Over the past 25
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`years, I have gathered significant experience designing and constructing CAR T
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`cells. I also have significant experience developing novel immunotherapies,
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`including CAR T-cell therapies.
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`8.
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`I received my BA magna cum laude in Biochemistry and Biophysics
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`from Harvard College in in 1971. I received my PhD in Molecular Biology and
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`Virology from University of California, Berkeley in 1975. I held a Postdoctoral
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`Fellowship in Chemistry and Chemical Engineering at the California Institute of
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`Technology between 1976 and 1979. I held a Postdoctoral Fellowship in Molecular
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`Biology at Roche Institute for Molecular Biology from 1979 to 1980. I received my
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`MD from the University of Miami School of Medicine in 1983. After completing
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`my residency in internal medicine at Georgetown University Hospital in 1986, I held
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`a Postdoctoral Fellowship there between 1985 and 1986. l then held a Postdoctoral
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`Fellowship in Medical Oncology at National Cancer Institute in Bethesda, Maryland
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`from 1986 to 1991. I was Chief Fellow between 1986 and 1988. I am currently
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`licensed to practice medicine in Massachusetts (License #74718).
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`9.
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`I was an Assistant Professor of Medicine at Harvard Medical School
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`from 1991 to 2003. I was a faculty member in the Department of Immunology at
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`Harvard Medical School from 1996 to 1998. I was an Assistant Adjunct Professor
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`in the Bioengineering Program at Boston University from 1996 to 2003. I was a
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`member of the Harvard Institute of Human Genetics (HIHF) at Harvard Medical
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`School from 1997 to 2003. I have been an Associate Professor of Medicine at Boston
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`University School of Medicine since 2003.
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`10.
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`I have been an active staff member at the following hospitals:
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`Deaconess Hospital (1991-1996); Dana-Farber Cancer Institute (1992-2002); Beth
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`Israel Deaconess Medical Center (1996-2003); Roger Williams Medical Center
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`(2004-2014); New England Baptist Hospital (2014-2015); Tufts Medical Center
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`(2015-2017); Cape Cod Hospital, Hyannis (locum tenens) (2017-2019); Morton
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`Hospital, Taunton (locum tenens) (2018-2019). I have been an active staff member
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`of Good Samaritan Medical Center, Brockton since 2019.
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`11.
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`I have held various administrative positions and committee
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`appointments over the years. I have served as Director of the Biotherapeutics
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`Development Lab in Boston, Massachusetts since 1992. I was a member of the
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`Biologics Therapy Group (1997-2003), Translational Research Committee (1998-
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`2003), and Cancer Center (1998-2003) at Beth Israel Deaconess Medical Center.
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`From 1999 to 2003, I was a member of the Experimental Therapeutics Program, GI
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`Malignancies Program, Breast Cancer Program, and Melanoma and Cutaneous
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`Oncology Program at the Dana Farber/Harvard Cancer Center. I was on the
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`Scientific Advisory Committee for the National Gene Vector Laboratory from 2003
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`to 2006. At Roger Williams Medical Center, I served as Chief of the Division of
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`Surgical Research (2004-2011), Tumor Board Coordinator (2004-2011), on the
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`Executive Committee of the Protocol Office (2005-2010), on the Protocol Review
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`Committee (2006-2014), on the Patent Committee (2006-2014), as Director of the
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`Cell Manipulation Core Facility (2011-2012), and on the Scientific Advisory Board
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`of the Cell Manipulation Core Facility (2011-2014). I served as Chief of the Section
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`of Hematology and Chair of Transfusion Safety Committee from 2014 to 2015 at
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`New England Baptist Hospital.
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`12.
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`I am a member of various professional societies, including the
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`American Association
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`for Cancer Research, American Association of
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`Immunologists, American Society for Clinical Oncology, American Society for
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`Hematology, the Gene Therapy Working Group of Harvard Medical School, and the
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`American Society of Gene Therapy. I am an ad hoc reviewer of various NIH Study
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`Sections, including Clinical Oncology, Small Business and Technology Transfer,
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`and Cancer Immunopathology and Immunotherapy.
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`13.
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`I have been a featured speaker on antibody and gene therapies in cancer
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`at national and international conferences. I have been the recipient of various awards,
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`including an American Association of Cancer Research Young Investigator Award
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`(1990), an American Cancer Society Clinical Oncology Career Development Award
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`(1993-1996), and a National Cancer Institute Research Career Development Award
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`(1996-2001). In 2003, I was recognized as an American Cancer Society Research
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`Scholar.
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`14.
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`I have established an on-campus cGMP facility for gene therapy
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`applications at Roger Williams Medical Center/Boston University School of
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`Medicine in Providence in 2007, with preparation and administration of >30 doses
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`of two different CAR-T cells for cancer therapies with full safety and indications of
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`efficacy in solid tumors.
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`15.
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`I have also been a consultant to government and private entities,
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`including major biotechnology companies. The topics of my consultancy include
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`biologic agents and gene therapy, including CAR T-cell therapy clinical trials.
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`16.
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`I have co-authored various publications on CAR T cells, including
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`those targeting disialoganglioside GD3 and carcinoembryonic antigen (CEA).
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`17.
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`I have initiated various clinical trials involving CAR T-cell therapy.
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`The clinical trials involved the use of CEA-specific CAR T cells for the treatment
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`of Adenocarcinoma, PMSA-specific CAR T cells for the treatment of prostate
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`cancer, and GD3-specific CART cells for the treatment of cancers involving tumors
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`derived from the neural crest cells, such as melanoma. I have secured FDA
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`Investigational New Drug (IND) status for 7 distinct antibody and CAR-T gene
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`therapy agents between 1999 and 2015, and one is currently pending.
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`III. MATERIALS CONSIDERED
`In forming the opinions in this declaration, I considered the materials
`18.
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`cited in this declaration, in light of the general knowledge in the art before December
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`9, 2011.
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`19.
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`I also relied on my education and knowledge, as well as my experience
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`in academic and clinical research projects, including those discussed in Section II.
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`20.
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`I reserve the right to amend and supplement any of the opinions in this
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`declaration as needed in response to new information and evidence made available
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`to me, additional analysis that leads me to conclude that supplementation is
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`necessary, new issues that may arise, any discovery, arguments, evidence, or
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`testimony presented in this inter partes review, and any Patent Trial and Appeal
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`Board (“Board”) decisions or orders.
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`IV. TECHNOLOGY BACKGROUND
`I understand that Petitioner contends that the ’140 patent cannot benefit
`21.
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`from the December 9, 2010 priority date and that the priority date should be
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`December 9, 2011. My opinion of the state of the art would not change if the Board
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`were to determine that the priority date is December 9, 2010.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`22. Below I provide a brief background of the state of the art before the
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`priority date to support my opinions on obviousness discussed in Sections X-XIII.
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`A person of ordinary skill in the art (“POSA”) would also have been aware of
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`additional references related to T cells and CARs.
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`A. T Cells
`23. The immune system comprises a diversity of distinct types of cells
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`including leukocytes, colloquially known as white blood cells. KENNETH MURPHY
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`ET AL., JANEWAY’S IMMUNOBIOLOGY 5 (7th ed. 2008) (Ex. 1029).1 Lymphocytes, a
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`type of leukocyte, contribute to the coordinated response to an invasion of
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`pathogens, such as microbes, into the body. Id. at 4-5.2 There are two major types of
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`lymphocytes: B cells and T cells. Id. at 9.
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`24. B cells produce antibodies. Id. at 9. Antibodies recognize and bind to
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`antigens that are present on pathogens, neutralize those pathogens, and assist in their
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`destruction and elimination from the body. Id. at 15, 28-29.
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`25. T cells do not produce antibodies. Id. at 9. Instead, on the surface of
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`T cells are receptors, called T cell receptors (“TCR”), which are similar to antibodies
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`1 I understand that the cited edition of Janeway’s Immunobiology is prior art to the
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`’140 patent under 35 U.S.C. § 102(b).
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`2 Citations refer to original pagination unless pagination is absent from the reference.
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`in that they also bind antigens. Id. The TCR includes surface receptors (α and β
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`chains) as well as associated proteins with intracellular signaling chains (such as
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`CD3), which together form a TCR complex. Id. at 9, 228-229. The antigen binding
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`portion of the TCR is similar to that found in the antigen binding portion of an
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`antibody. Id. at 15. Other cell surface proteins, namely the cell surface receptors
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`CD4 and CD8, associate with the TCR complex. Id. at 31. T cells with CD4 receptors
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`are identified as “helper” T cells and T cells with CD8 receptors are “cytotoxic” T
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`cells. Id. 31-32; 350. Cytotoxic T cells kill other cells. Id. 33-34; 350. Helper T cells
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`serve to activate B cells to secrete antibodies and activate cytotoxic T cells to kill
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`other cells. Id. 32-34; 350.
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`26. A TCR can recognize an antigen when the antigen is bound to the
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`surface of another type of cell referred to as an antigen-presenting cell (“APC”). Id.
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`at 7. The antigen is presented on the APC by a major histocompatibility (“MHC”)
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`molecule. Id. at 17-18; Figure 1.16. Upon recognition and binding with the antigen-
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`MHC complex, the T cell becomes activated and secretes certain chemicals, such as
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`cytokines, that recruit other cells in the immune system to coordinate the immune
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`response against the threat represented by the antigen. Id. at 351, Figure 8.27. Below
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`is an illustration of the basic structural components of the TCR-MHC interaction,
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`which was known before the priority date:
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`Antigen-Presenting cell
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`MHC
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`4-18B
`Ligand
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`CD3~
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`Tcell
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`■ 4-l BB
`■ CD3~
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`Antigen
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`■ Antigen-binding domain
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`See also id.at 229, Figure 6.10; 347.
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`-
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`-
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`Cell membrane
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`
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`27. As shown in the figure above in paragraph 26, the TCR, through its
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`antigen-binding domain, recognizes and binds to an antigen presented on an APC by
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`an MHC complex. See id. at 7. This binding mechanism then triggers T cell
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`activation. T cell activation involves two distinct signaling events known as a
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`primary signal and a costimulatory signal. Id. at 323-325; 344.
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`28. The primary signal is mediated by the TCR complex, which is depicted
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`in the left-hand side of the figure in paragraph 26. See id. at 349, Figure 8.25. An
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`important part of this primary signaling cascade relies upon the CD3 zeta, or CD3-
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`ζ, domain or chain. Id. at 228-29. As depicted in the figure of paragraph 26 in
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`Declaration of Richard P. Junghans, Ph.D., M.D.
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`orange, the CD3 zeta signaling domain is located in the intracellular region of the
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`TCR. Id. at 228-229. A POSA would have known before the priority date that the
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`CD3 zeta domain is important for T-cell activation.
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`29. The costimulatory signal is mediated by other receptors that span the T
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`cell surface, are in proximity to the TCR complex, and also physically interact with
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`the APC via a separate binding site. Id. at 346-347. As illustrated in the figure of
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`paragraph 26 in green, one such costimulatory receptor was the 4-1BB receptor (also
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`known as “CD137”). Id. at 347. A POSA also understood that recruitment of the
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`costimulatory signal led to increased T cell activation and increased cytokine
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`secretion.
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`30. The cascade of signaling events that is triggered by the binding of the
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`TCR to the MHC and that ultimately leads to the T cell’s secretion of chemicals
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`including cytokines was therefore understood before the priority date. See id. 324,
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`358-359, Figure 8.32.
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`B. Chimeric Antigen Receptor T Cells (CAR T Cells)
`31. The body’s natural ability to recognize and destroy cancer cells via
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`T-cell activation has led to the development of immunotherapies for cancer
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`treatment. One type of immunotherapy uses chimeric antigen receptor modified
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`T cells, referred to as CAR T cells.
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`32. The CAR is a chimera, or fusion, of several protein domains, that
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`include three basic, yet essential, components: (1) an antigen-binding domain, which
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`usually consists of a single-chain fragment of variable region (“scFv”) antibody that
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`recognizes a tumor-associated antigen, followed by a hinge region for proper
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`extracellular display of the scFv; (2) an intracellular signaling domain; and (3) a
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`transmembrane domain that spans the T cell membrane to fuse together the scFv
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`antibody to the intracellular signaling domain. An example of how a TCR and CAR
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`are positioned on the T cell surface and their interactions with an antigen present on
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`the cell surface of another cell are shown below. As depicted, the TCR of a T cell
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`binds to an antigen on the surface of an APC. A CAR expressed on a T cell, on the
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`other hand, can bind directly to an antigen on a tumor cell.
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`Antigen-Presenting cell
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`Tumor cell
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`MHC
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`4-lBB
`Ligand
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`Tumor antigen
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`CD3~
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`TCR
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`Antigen
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`T cell
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`CAR
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`Cell membrane
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`■ Antigen-binding domain
`■ Transmembrane domain
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`■ 4-18B
`■ CD3~
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`33. Before the priority date, use of scFv antibodies in CARs was already
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`known in the art. It was understood that an scFv antibody comprised the “heavy”
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`and “light” variable portions of the antibody, which provide the antibody its binding
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`specificity against a particular antigen. Ex. 1029 at 112. An antibody, which consists
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`of two heavy chains and two light chains that form a “Y” shape, also includes
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`“constant” regions (gray, below). Id. at 111-112. A simplified illustration of the
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`location of the heavy (red, below) and light (blue, below) variable regions on an
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`antibody and as an scFv binding domain of a CAR is below:
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`34. For example, it was known at the time before the priority date that in
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`hematological or blood cancers including lymphoma and leukemia, the cancerous or
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`malignant cells originate from lymphocytes, such as B cells. Id. at 308-311,
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`Figure 7.41. Hematological cancers are cancers that affect the blood, bone marrow,
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`and lymph nodes. It was also understood that the surface receptor identified as CD19
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`was expressed on the cell surface of B lineage cells,3 including malignant B cells.
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`Id. at 314, Figure 7.45. Accordingly, a POSA understood that one way to direct a T
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`cell response to these malignant B cells was by engineering T cells to recognize and
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`bind to the CD19 receptor on the malignant B cells, thereby causing T cell activation
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`3 B cell lineage refers to the development of different types of B cells from a common
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`ancestor; specifically, a common lymphoid progenitor cell. Ex. 1029 (Janeway) at
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`4, 309, 314.
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`and targeted destruction of those malignant B cells. Unlike naturally occurring T
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`cells, which require APCs to present antigens that TCRs can bind to, the genetically
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`modified T cells can bind to CD19 receptors directly on the surface of malignant
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`B cells. B cell malignancies wherein the malignant B cells express CD19 are
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`commonly referred to as CD19+ B cell malignancies. As I discuss infra, in
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`Section VIII, others had already constructed CARs with an scFv antibody targeting
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`CD19+ B cells for use as a CAR T cell therapy before the priority date.
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`C. Engineering CAR T Cells
`35. To administer CAR T-cell therapy, one first obtains normal T cells
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`circulating in a cancer patient’s blood through a technique called leukapheresis.
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`Then gene transfer techniques are used to insert a CAR into a patient’s T cells to
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`genetically modify those T cells ex vivo.
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`36. These gene transfer techniques, such as using a viral vector, were well
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`known in the art before the priority date. Specifically, a POSA knew that certain
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`types of viruses, such as retroviruses and lentiviruses, could be manipulated such
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`that those viruses included the genetic coding sequence of the CAR construct. The
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`virus is then utilized as a vehicle or vector to “infect” or transduce the CAR-encoding
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`genetic sequence into T cells. The cellular machinery in the CAR-transduced T cells
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`then transcribes the CAR genetic sequence into mRNA and then translates the
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`mRNA into the CAR protein, which becomes inserted, or expressed, on the cell
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`surface of the T cell.
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`37. The genetic sequences included in the viral vector are derived from
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`plasmids, which are circular double stranded DNA molecules. Ex. 1029 at 682,
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`Figure 15.20. These plasmids include the DNA sequence encoding the protein
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`domains that make up the CAR construct. The CAR construct is engineered by
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`“stitching” together the DNA sequences that encode for the various protein domains
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`comprising the CAR construct. See, e.g., infra paragraph 51. The CAR construct is
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`then inserted into a plasmid along with other gene sequences (e.g., green, blue, and
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`orange, below) necessary for transduction into the T cells and expression of the CAR
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`protein:
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`CAR construct
`sequence
`
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`38. Once the viral vectors comprising the plasmid have transduced T cells
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`with the sequence encoding the CAR construct, the T cells will then translate the
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`genetic sequence into an amino acid sequence that will undergo modification to form
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`the CAR T receptor that becomes inserted into the cell surface of the T cell.
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`Specifically, certain cellular machinery will transcribe the DNA sequence into an
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`mRNA sequence and that mRNA sequence will be translated into a chain of amino
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`acid residues bound together to form a polypeptide chain. Alberts et. al.,
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`MOLECUL