Vol.
`
`1, 965-972,
`
`September
`
`1995
`
`Clinical
`
`Cancer
`
`Research
`
`965
`
`of Serotherapy
`I Clinical
`Phase
`Trial
`an Immunoglobulin
`Leukemia
`with
`Antibody
`to CD15’
`
`in Patients
`M Monoclonal
`
`with Acute Myeloid
`
`Edward
`Roger
`
`Kathy
`D. Ball,2
`Herzig,
`Laura
`
`Selvaggi,
`Clark,
`Vicki
`
`David
`Malley,
`
`Hurd,
`
`Jeannette
`
`Margarida
`
`Persichetti,
`
`and
`
`deMageihaus-Silverman
`
`Transplantation,
`Marrow
`of Hematology/Bone
`Division
`Pennsylvania
`Pittsburgh,
`Medical
`Center,
`Pittsburgh
`K. S., L. C., V. M.,
`J. P., M. D. S.]; Bowman-Gray
`Winston-Salem,
`North
`Carolina
`27103
`[D. H.];
`Louisville,
`Louisville,
`Kentucky
`40292
`[R. H.]
`
`of
`University
`15213
`[E. D. B.,
`Medical
`Center,
`and University
`of
`
`ABSTRACT
`
`used
`
`or
`
`INTRODUCTION
`of malignancies
`treatment
`the
`in
`been
`mAbs
`have
`necessary
`for
`is
`It
`antigens.
`expressing
`tumor-associated
`as
`the
`ability
`such
`mechanism,
`mAb
`to
`have
`an
`effector
`antibody-dependent
`cellu-
`or mediate
`activate
`complement
`(C’)
`agent,
`such
`a radio-
`as
`a cytotoxic
`lar
`cytotoxicity,
`to carry
`or
`of
`tumor
`mAbs
`cells.
`isotope,
`drug,
`toxin,
`to allow
`for
`killing
`that
`to
`cell
`lead
`which
`regulate
`higand/receptor
`interactions
`stimulate
`programmed
`death
`through
`indirect
`mechanisms
`or
`be used
`therapeutically.
`cell
`death
`(such
`as
`apoptosis)
`could
`also
`trials
`of
`in vivo mAb
`In one
`of
`the
`early
`and most
`encouraging
`therapy,
`a
`patient
`with
`B-cell
`non-Hodgkin’s
`lymphoma
`
`the
`to
`
`(1).
`
`bone
`
`bone
`(2,
`
`years
`in patients
`labeled
`reductions
`significant
`was
`transplant.
`marrow
`marrow
`and
`liver,
`and
`marrow,
`researchers
`3). Many
`mAbs
`to reduce
`toxic-
`cellular
`cytotoxicity
`
`after
`with
`with
`of
`
`in
`
`Sixteen
`treated
`
`(AML)
`heukemia
`acute myeloid
`with
`patients
`PM-81,
`of mAb
`i.v.
`infusion
`a continuous
`with
`were
`differentiation
`the
`cellular
`directed
`against
`an
`mAb
`1gM
`cells
`of >95%
`on leukemia
`which
`is expressed
`CD15,
`antigen
`as
`referred
`to
`PM-8i,
`also
`with
`AML. MAb
`of
`patients
`human
`and rabbit
`comphe-
`is capable
`of activating
`MDX-ii,
`AML
`cells.
`In this
`Phase
`lysing
`CD15-positive
`and
`ment
`with
`0.5,
`i.0,
`or
`1.5 mg/kg
`were
`treated
`patients
`study,
`followed
`by conven-
`over
`a 24-h
`delivered
`MDX-ii
`period
`decreases
`circulating
`chemotherapy.
`Transient
`tional
`ob-
`chemotherapy)
`were
`to
`cells
`postinfusion
`(prior
`blast
`to show MDX-ii
`binding
`able
`at all doses. We were
`served
`patients
`who
`achieved
`stable
`to bone marrow
`blasts
`in those
`Serum MDX-il
`was detectable
`serum
`levels
`of MDX-ii.
`the
`and
`1.5-mg/kg
`doses.
`Doses
`of
`0.5
`and
`i.0 mg/kg
`i.0-
`were
`generally
`well
`tolerated,
`with no toxicities
`greater
`than
`grade
`II
`(Eastern
`Group)
`reported.
`Cooperative
`Oncology
`However,
`two
`of
`five
`patients
`receiving
`the
`1.5-mg/kg
`dose
`experienced
`grade
`IV toxicities
`that
`resolved
`with treatment
`(one
`of
`patients
`completed
`the
`infusion).
`Common
`these
`toxicities
`reported
`included
`fever,
`chills,
`and
`hypotension.
`one
`patient
`developed
`human
`antimouse
`antibodies
`Only
`4 weeks
`posttreatment.
`that
`This
`determined
`study
`a biologically
`effective
`adminis-
`be
`that
`can
`dose
`mg/kg
`is
`tered
`safely with little
`toxicity.
`we are
`on these
`results,
`Based
`of MDX-11
`infusion
`following
`pursuing
`a Phase
`I/LI
`study
`chemotherapy
`for patients
`with
`relapsed
`AML.
`
`in
`
`I
`
`at
`
`at
`1.0
`
`cells.
`for
`
`achieved
`treatment
`AML3
`iodine-i31
`tumor
`enough
`Whole-body
`biopsies
`spleen
`have
`ities
`various
`
`as
`also
`and
`
`We
`
`agent
`formed
`in
`relapse
`
`or
`
`to
`
`several
`for
`lasting
`remission,
`complete
`Studies
`mAb
`with
`an antiidiotype
`with
`CD33)
`(reactive
`using
`the mAb M195
`in transient
`success
`have
`also met with
`cytoreduction
`some
`cases,
`In
`to
`bone
`proceed
`to
`patients
`radiolocalization
`for
`imaging
`uptake
`in
`the
`Mi95
`showed
`1 h after
`infusion
`as
`early
`chimeric
`and
`humanized
`used
`antibody-dependent
`promote
`(4-6).
`malignancies
`(anti-
`PM-81
`mAb
`the
`described
`have
`previously
`reactive
`1, which
`is
`to as MDX-1
`subsequently
`referred
`CD15),
`with
`AML.
`The
`>95%
`of
`patients
`leukemia
`cells
`from
`with
`is
`a
`trisaccharide
`recognized
`by
`anti-CD15
`antibodies
`epitope
`(also
`referred
`to as
`the
`within
`pentasaccharide
`LNF-III
`structure
`on
`found
`neutrophils,
`CD15
`x,
`Lewis
`or
`SSEA-1).
`is
`LNFP-III,
`in embryonic
`tissues,
`is present
`and
`and monocytes,
`eosinophils,
`is
`Thus,
`MDX-li
`leukemias.
`and myeloid
`adenocarcinomas,
`1 is
`and monocytes.
`MDX-1
`granulocytes
`normal
`reactive
`with
`the
`colony-forming
`unit-gran-
`with
`lymphocytes
`not
`reactive
`unit-erythroid
`8).
`This
`(7,
`or
`burst-forming
`ulocyte-monocyte
`both
`activating
`rabbit
`and
`of
`1gM antibody
`capable
`mAb
`is
`an
`MDX-1
`1 is cytotoxic
`to early
`human
`complement
`(9). Although
`malignant
`myeloid
`leukemia
`precursors
`and
`some
`normal
`my-
`eloid
`precursors,
`pluripotent
`stem
`cells
`are
`unaffected
`(9,
`10).
`Studies
`exploring
`the
`use
`of MDX-1
`1 as
`a
`therapeutic
`per-
`are
`in progress.
`An
`early
`Phase
`I clinical
`trial
`was
`in which
`three
`patients
`with
`AML
`with
`resistant
`disease
`were
`given
`up
`500
`mg MDX-1
`1 in
`All
`vivo.
`
`5/15/95;
`in
`
`was
`
`3/23/95;
`Received
`project
`1 This
`Cancer
`National
`(Bethesda,
`Services
`2 To whom
`requests
`Hematology/Bone
`Medical
`Center,
`
`200
`
`revised
`supported
`Institute,
`MD)
`for
`Marrow
`Lothrop
`
`accepted
`part
`by
`Department
`NIH,
`by Medarex,
`and
`reprints
`should
`Transplantation,
`Street,
`Pittsburgh,
`
`5/30/95.
`Grant
`
`the
`from
`CA31888
`Human
`of Health
`NJ).
`(Annandale,
`Inc.
`at Division
`addressed,
`of Pittsburgh
`University
`PA 15213.
`
`and
`
`of
`
`are: AML,
`used
`abbreviations
`3 The
`transplantation;
`autologous
`bone marrow
`HAMA,
`human
`normal
`human
`serum;
`phosphate-buffered
`saline/bovine
`serum
`III,
`lacto-N-fucopentaose
`III; ECOG,
`Group.
`
`acute myeloid
`gam,
`goat
`antimouse
`albumin/sodium
`Eastern
`Cooperative
`
`leukemia;
`antimouse;
`antibodies;
`azide;
`
`ABMT,
`NHS,
`PBA,
`LNFP-
`Oncology
`
`be
`
`
`
`Downloaded from on August 31, 2015. © 1995 American Association for Cancerclincancerres.aacrjournals.org
`
`
`Research.
`
`PETITIONER'S EXHIBITS
`
`Exhibit 1010 Page 1 of 9
`
`

`
`966
`
`Phase
`
`I Clinical
`
`Trial
`
`of
`
`Serotherapy
`
`in Patients
`
`with
`
`AML
`
`in
`decrease
`25% transient
`approximate
`an
`experienced
`patients
`also
`1 has
`(1 1). MDX-1
`with
`little
`toxicity
`blast
`counts
`leukemic
`for
`mAb
`(AML-2-23),
`an
`anti-CD14
`along
`with
`been
`used,
`ex
`with
`for ABMT
`for
`patients
`marrow
`bone
`treatment
`vivo
`study
`have
`been
`encouraging
`of
`results
`AML
`(12,
`13).
`planned.
`trial
`is being
`and
`a randomized
`Despite
`successes
`with
`ABMT
`in the
`
`of
`The
`
`this
`
`treatment
`
`of AML,
`
`it
`
`for
`
`that
`up
`to
`note
`is
`important
`implying
`relapse,
`eventually
`transplant
`patients
`prepare
`the
`of
`while
`most
`And
`in
`shown
`have
`leukemias
`to date
`of
`tumor
`burden,
`few cases
`been
`reported
`(14,
`15). With
`limited
`toxicities
`associated
`evaluating
`the
`of MDX-1
`role
`ment
`of AML
`adjunct
`an
`as
`immunotherapy.
`post-ABMT
`study
`were
`to establish
`cal dose
`of MDX-1
`1. The
`with
`relapsed
`or
`secondary
`MDX-11
`using
`a continuous
`conventional
`chemotherapy.
`
`safety,
`study
`
`to
`
`of
`
`with
`therapies
`all
`leukemia
`
`transplants
`to
`used
`cells.
`
`patients
`50%
`ablative
`the
`that
`to remove
`fail
`mAb
`vivo
`used
`serotherapies
`reductions
`in transient
`promise
`remissions
`or partial
`complete
`because
`in mind,
`and
`this
`mAb
`serotherapies,
`with
`1 mAb
`serotherapy
`to standard
`chemotherapy
`this
`The
`objectives
`of
`feasability,
`optimal
`and
`design
`treating
`included
`with
`escalating
`AML
`infusion
`over
`i.v.
`
`for
`
`of
`we
`
`in
`of
`have
`the
`are
`treat-
`the
`and
`for
`Phase
`biologi-
`patients
`doses
`
`I
`
`of
`
`by
`
`24
`
`h followed
`
`MATERIALS
`mAb
`
`AND
`
`METHODS
`
`for
`
`at
`
`MDX-!
`use
`clinical
`IND
`tration
`bioreactor
`fiber
`Supernatant
`of
`the
`end
`Final
`purified
`HPLC
`and
`binding
`and
`assays,
`and
`endotoxin
`safety.
`
`and
`filtered
`is
`purification
`antibody
`gels
`SDS
`specificity
`for
`general
`bevels,
`performing
`
`viral
`
`and
`
`prepared
`NJ) was
`Inc., Annandale,
`1 (Medarex,
`and Drug
`Adminis-
`under
`the United
`States
`Food
`using
`a
`hollow
`4362.
`MDX-11
`is manufactured
`using
`HPLC.
`purified
`by
`ion
`exchange
`process
`and
`during
`the manufacturing
`by
`column.
`following
`buffer
`exchange
`control
`using
`is monitored
`for
`quality
`activity
`using
`identity
`purity,
`for
`for
`and
`assays
`(flow
`cytometry)
`and
`cytotoxicity
`safety
`by
`determining
`DNA
`levels,
`testing,
`general
`animal
`
`HL6O Cells
`cell
`leukemia
`a CD15-positive
`HL6O,
`Culture
`Collection
`the American
`Type
`from
`in RPMI
`1640
`cells
`were
`cultured
`The
`Island,
`NY)
`with
`Technologies,
`Grand
`Logan,
`UT),
`L-glutamine,
`penicillin,
`tamicin.
`
`and
`
`was
`line,
`(Bethesda,
`(GIBCO-BRL,
`10%
`FCS
`streptomycin,
`
`obtained
`MD).
`Life
`(Byclone,
`and
`gen-
`
`Serum
`
`MDX-!!
`
`Determination
`
`sandwich
`patient
`
`a
`using
`determined
`were
`1 levels
`MDX-i
`Serum
`phase,
`solid
`in the
`gam-IgM
`incorporated
`which
`ELISA
`phosphatase-conju-
`alkaline
`step,
`and
`intermediate
`the
`sera
`Blood
`samples
`antibody.
`final
`labeled
`the
`gam-IgM
`as
`gated
`before,
`during,
`patients
`at
`screening,
`and
`obtained
`from
`were
`coated
`overnight
`Briefly,
`96-well
`plates
`were
`after
`infusion.
`and
`and
`blocked
`with
`at 4#{176}Cwith
`1 p.g gam-IgM,
`washed
`with
`PBS,
`5% PBA for 45 mm to 2 h at
`curve
`dilutions
`of
`37#{176}C.Standard
`1gM (30-0.003
`mg/mI)
`were
`made
`in
`10% NBS.
`Patient
`sera
`were
`diluted
`in 1% PBA;
`higher
`dilutions
`of patient
`serum
`
`as
`
`1:10
`
`in
`
`washed
`were
`Plates
`in PBA.
`diluted
`10% NHS
`made
`were
`curve,
`standard
`pA of
`negative
`control,
`and
`100
`blocking,
`after
`wells.
`appropriate
`were
`added
`to the
`dilutions
`serum
`patient
`and
`washing,
`37#{176}Cfor
`1.5
`to
`3 h. After
`incubated
`at
`were
`Plates
`to
`for
`1.5
`gam-IgM
`was
`added
`phosphatase-conjugated
`alkaline
`washed
`and
`developed
`with
`then
`was
`37#{176}C.The
`plate
`3 h at
`The
`reaction
`was
`stopped
`disodium.
`p-nitrophenyl
`phosphate
`20 mm,
`and
`plates
`were
`after
`with
`1 N sodium
`hydroxide
`MR660)
`using
`a 410A
`filter,
`read
`in an ELISA
`reader
`(Dynatech
`wells.
`Standard
`curve
`was
`blanking
`on
`the
`negative
`control
`performed
`in duplicate,
`patient
`samples
`in triplicate.
`This
`assay
`
`the
`
`to
`
`was
`above
`
`able
`0.5
`
`detect
`p.g/ml
`
`levels
`were
`
`as
`able
`
`low
`as
`to be
`
`0.1
`(cid:1)ig/ml;
`quantified.
`
`levels
`
`of MDX-11
`
`Serum
`
`CD15 Determination
`study
`from
`patients
`in this
`Sera
`which
`a
`using
`blocking
`assay
`CD15
`Rosedale,
`Glycosystems,
`Inc.,
`(Oxford
`before
`infusion
`obtained
`from patients
`were
`infusion).
`The
`CD15
`antigen-positive
`cell
`vested,
`washed,
`resuspended
`to 2 X i07/ml
`on
`ice
`until
`blocking.
`The
`part
`of
`the
`
`assayed
`were
`incorporated
`NY).
`Blood
`and
`at
`t
`line HL6O
`in PBA,
`assay
`was
`
`=
`
`soluble
`for
`LNFP-III
`samples
`
`24
`was
`and
`carried
`
`h (end
`har-
`stored
`out
`
`first
`
`1 was
`were
`in
`
`in
`
`1 year
`
`and
`
`in
`
`of
`
`dark
`
`of
`
`MDX-i
`plates.
`round-bottomed
`96-well
`in
`duplicate
`units)
`10 pA (0.0075
`in PBA,
`units/ml
`diluted
`to 0.75
`and
`2.0 mg/ml
`of LNFP-III,
`added
`to each well.
`A standard
`curve
`to the microtiter
`PBS,
`was made
`by
`adding
`0.5-10
`(cid:1)i.l LNFP-III
`and NHS,
`10 (cid:1)il
`wells
`containing
`MDX-1
`1. For
`patient
`samples
`to wells
`con-
`of
`undiluted,
`and
`1:10
`dilutions
`were
`added
`1:5,
`to
`20
`p.1 with
`were
`taming
`MDX-1
`1. Final
`volumes
`brought
`PBA.
`For negative
`controls,
`1gM (Coulter,
`irrelevant
`5 (cid:1)i.l
`mouse
`Hialeah,
`FL)
`and
`15
`p.1 PBA
`were
`added
`to the MDX-1
`1. The
`was
`plate
`then
`incubated
`at
`room
`temperature
`with
`shaking
`for
`30 mm. After
`15 mm,
`receptors
`on HL6O
`cells were
`blocked
`Fc
`by
`adding
`#{189}volume
`human
`blocking
`IgG (10 mg/ml)
`and
`of
`incubating
`for
`10
`to
`15 mm at 4#{176}C(this
`incubation
`coincided
`end
`with
`the
`the
`antigen/antibody
`binding
`incubation).
`After
`the
`incubations,
`75
`p.1 (106)
`Fc-blocked
`HL6O
`cells were
`added
`to each
`microtiter
`well.
`The
`plate
`was
`incubated
`for
`60 mm at
`4#{176}C.The
`contents
`of
`the microtiter
`wells
`were
`then
`transferred
`to appropriately
`labeled
`tubes.
`All
`tubes
`were
`washed
`with
`PBA
`twice,
`and
`25 p.1 FITC-labeled
`gam-IgM
`were
`added
`to each
`cell
`dark
`pellet
`and
`incubated
`in the
`for
`30
`to 40 mm at 4#{176}C.After
`were
`washing,
`the
`cell
`pellets
`resuspended
`in
`0.5
`ml
`1%
`paraformaldehyde
`and
`allowed
`to fix
`in the
`at 4#{176}Cfor
`1 h.
`Samples
`were
`analyzed
`by
`flow
`cytometry
`using
`histogram
`overlays
`with
`appropriate
`controls.
`Standards
`curves
`of
`r2
`0.98 were
`used
`for
`determining
`the
`amount
`soluble
`CD15
`patient
`serum.
`Patients
`relapsed
`
`in first
`who were
`were
`treated
`with
`
`than
`greater
`remission
`1-(cid:1)3-D-arabinofuranosylcytosine
`
`then
`(100
`daunorubicin
`days
`and
`7
`for
`i.v.
`mg/m2/day)
`regimen.
`induction
`an
`as
`days
`3
`i.v.
`for
`day)
`in second
`or were
`1 year
`than
`in less
`relapsed
`1-(cid:1)3-D-arabinofuranosylcytoside
`high-dose
`received
`12 doses
`unless
`they
`had
`12 h)
`iv.
`for
`every
`regimen.
`Etoposide
`(100
`mg/m2/day)
`this
`and mitoxantrone
`(10 mg/m2/day)
`given
`
`(45
`Patients
`third
`
`or
`
`mg/m2/
`who
`relapse
`g/m2
`with
`5 days
`were
`
`(3
`treatment
`i.v.
`for
`5 days
`
`prior
`given
`iv.
`
`for
`
`
`
`Downloaded from on August 31, 2015. © 1995 American Association for Cancerclincancerres.aacrjournals.org
`
`
`Research.
`
`PETITIONER'S EXHIBITS
`
`Exhibit 1010 Page 2 of 9
`
`(cid:18)
`

`
`used
`mens
`
`in patients
`listed.
`
`who
`
`had
`
`already
`
`been
`
`treated
`
`with
`
`prior
`
`regi-
`
`of Total
`
`CD15-positive
`
`Cells
`
`and Cell-
`
`by
`1
`I
`
`a
`
`the
`
`on
`
`(cid:1)i.l
`
`and
`
`Determination
`bound MDX-!!
`determined
`cells was
`CD15
`of positive
`Total
`percentage
`using MDX-i
`cells
`patient
`the
`staining
`for CD15
`on
`indirect
`Cell-bound
`MDX-1
`antibody.
`secondary
`FITC-labeled
`and
`a
`to
`antibody
`directly
`secondary
`the
`was
`determined
`by
`using
`cell
`surface.
`Blood
`the
`bound
`to
`stain
`for MDX-1
`1 already
`CD15-positive
`and
`cells
`of
`total
`samples
`for
`determination
`the
`patients
`screening
`at
`taken
`from
`cell-bound
`MDX-1
`1 were
`(b one marrow),
`infusion.
`In addition,
`before,
`during,
`and
`after
`bone marrow
`sample
`was
`taken
`for
`determination
`of cell-bound
`MDX-1
`1 at
`the
`the
`termination
`of
`infusion.
`Leukemic
`cells were
`h
`isolated
`by Ficoll-Hypaque
`gradient
`centrifugation
`within
`12
`of
`collection.
`Separated
`were
`kept
`in RPM!
`1640
`ice
`cells
`until
`ready
`use.
`Prior
`to assay
`cells
`were
`washed
`with
`cold
`for
`PBA
`and
`resuspended
`to 2 X i07/ml
`in PBA.
`Each
`assay
`tube
`received
`106
`(50
`pA)
`cells.
`For
`determination
`of
`total
`CD15-
`positive
`cells,
`cells
`were
`blocked
`with
`25
`human
`IgG
`(10
`mg/ml)
`for
`15 mm on
`ice. Autofluorescence
`control
`tubes
`re-
`25
`ceived
`25
`pA PBA,
`negative
`control
`tubes
`received
`pA irrel-
`evant
`1gM,
`positive
`control
`tubes
`received
`25
`gil W632
`super-
`natant
`(anti-HLA),
`and
`test
`tubes
`(assayed
`in duplicate)
`received
`25
`(cid:1)al MDX-1
`1 (200
`p.g/ml).
`Cells
`were
`incubated
`with
`primary
`antibody
`for
`60 mm on
`ice
`and washed.
`For
`cell-bound
`MDX-1
`1 determination
`and after washing
`following
`incubation
`for
`of
`primary
`antibody
`total
`CD1S-positive
`cells,
`50
`p.1 PBA
`were
`added
`to
`autofluorescence
`tubes,
`50
`of
`F(ab’)2
`(cid:1)il
`gam-IgG
`1gM (H
`+
`L)
`FITC-labeled
`were
`added
`to
`the
`+
`remaining
`tubes
`and
`incubated
`for
`30 mm on
`ice
`(dark).
`Cells
`in
`were
`washed,
`resuspended
`1.0 ml
`1% paraformaldehyde,
`stored
`in
`the
`dark
`at
`4#{176}C,and
`allowed
`to
`fix
`for
`1 h before
`analysis
`by
`flow
`cytometry.
`For
`cell-bound
`MDX-1
`1 determi-
`nation,
`staining
`with
`FITC-labeled
`secondary
`antibody
`occurred
`simultaneously
`with
`the
`incubation
`of
`the
`primary
`antibody
`total
`CD15-positive
`cells.
`The
`negative
`control
`used
`for
`bating
`cell-bound
`MDX-1
`1 was
`the
`highest
`of
`either
`the
`MDX-
`1 1 or
`the
`autofluorescence
`tube
`prepared
`cell-bound
`that
`time
`point.
`
`for
`calcu-
`t
`
`=
`
`0
`for
`
`HAMA
`
`the
`
`Determination
`a sand-
`using
`assayed
`were
`antibodies
`Human
`antimouse
`with MDX-li.
`coated
`was
`the
`plate
`ELISA
`in which
`wich
`step
`and
`goat
`intermediate
`the
`serum
`was
`incubated
`as
`Patient
`final
`antibody.
`was
`the
`alkaline
`phosphatase
`IgG
`antihuman
`at 4#{176}Cwith
`1 p.g/well
`were
`coated
`overnight
`plates
`Briefly,
`5% PBA
`blocked
`with
`washing,
`the
`plate was
`1. After
`of MDX-1
`sera
`and NHS
`(negative
`for
`1 to 2 h at 37#{176}Cand washed.
`Patient
`1:40,
`1:80,
`and
`1:160
`control)
`were
`diluted
`in duplicate
`1:20,
`from
`individual
`who
`using
`1% PBA.
`Positive
`control
`serum
`repeated
`treatment
`with
`exhibited
`a HAMA
`titer
`of
`1:240
`(after
`and
`an anti-Fc
`recep-
`a bispecific
`antibody
`consisting
`of PM-81
`One
`hundred
`of
`all
`tor
`antibody)
`was
`diluted
`to 1:20-1:480.
`wells
`received
`100
`pi
`dilutions
`were
`added
`to the
`plates;
`blank
`1% PBA.
`The
`plate was
`incubated
`for
`1.5 h at 37#{176}Cand washed.
`A working
`dilution
`of goat
`antihuman
`IgG alkaline
`phosphatase
`
`an
`
`ml
`
`Clinical
`
`Cancer
`
`Research
`
`967
`
`added
`was
`washing,
`mm at
`NaOH,
`
`the
`room
`and
`
`to
`
`all wells
`substrate
`temperature.
`the
`plate
`
`and
`was
`
`was
`
`incubated
`added
`and
`The
`reaction
`read
`at 405
`
`h at 37#{176}C.After
`1.5
`for
`to develop
`for
`20
`allowed
`with
`stopped
`1 N
`was
`nm within
`20 mm.
`
`Flow
`
`Cytometry
`
`using
`performed
`was
`cytometry
`Flow
`acquiring
`For
`II software.
`with
`Lysis
`FACScan
`scatter
`forward
`patients,
`cells
`from
`leukemia
`FITC-labeled
`fixed,
`were
`standardized
`with
`settings
`standardized
`using
`and
`fluorescence
`settings
`were
`and
`the
`bead
`mixture.
`HL6O
`cells
`Both
`Typically,
`were
`provided
`by Medarex,
`Inc.
`All
`acquired
`for
`each
`sample.
`on-site
`analyses
`by Medarex,
`Inc.,
`for
`consistency.
`
`Dickinson
`a Becton
`antibody-stained
`and
`side
`scatter
`HL6O cells,
`a
`standard
`bead
`mixture
`cells
`were
`monitored
`
`standard
`10,000
`were
`
`Study
`
`Design
`Eligibility.
`Patient
`of
`relapsed
`subclass
`British
`phase
`or
`myelodysplastic
`eligibility
`eligible.
`Age
`Karnofsky
`performance
`kemic
`blast
`cells were
`1 1 . The
`criteria
`for
`were
`delineated
`as
`expiratory
`volume
`
`Patients
`AML
`prior
`included
`status
`required
`cardiopulmonary,
`left
`ventricular
`(one
`second)
`
`of
`and
`cytotoxic
`patients
`required
`was
`to be >20%
`liver,
`ejection
`>70%
`
`French-American-
`any
`secondary
`AML
`after
`drug
`therapy
`were
`18
`years
`or
`older.
`to be
`>70%.
`Leu-
`positive
`for MDX-
`and
`renal
`function
`fraction
`>0.4,
`forced
`predicted,
`hepatic
`
`a
`
`transaminases
`ance
`>50
`
`<
`
`three
`
`times
`
`normal
`
`values,
`
`and
`
`creatinine
`
`clear-
`
`mb/h.
`hydrated
`were
`Patients
`Administration.
`mAb
`of MDX-1
`to administration
`for
`4 h prior
`saline
`mal
`in hopes
`of minimizing
`allergic
`volume
`intravascular
`Patients
`were
`also
`premedicated
`with
`and/or
`toxicities.
`acetaminophen.
`The mAb
`was
`administered
`hydramine
`and
`a peripheral
`central
`line.
`Three
`patients
`or
`through
`either
`of 0.5 mg/kg,
`8 patients
`received
`1.0 mg/kg,
`ceived
`doses
`patients
`received
`1.5 mg/kg.
`The
`infusion
`rate was
`10 mb/h
`
`nor-
`with
`1 to increase
`reactions
`diphen-
`i.v.
`re-
`5
`
`and
`(240
`
`imme-
`
`ml/24
`diately
`
`sion,
`Serum
`uric
`and
`infusion.
`counts
`1 1,
`total
`These
`MDX-i
`cytology,
`screening
`blood
`sion.
`try,
`done
`assays
`
`conventional
`chemotherapy
`received
`h).
`Patients
`of
`the MDX-i1
`infusion.
`completion
`following
`Before
`and during
`Tests
`Performed.
`1 infu-
`the MDX-1
`counts
`and
`electrolytes
`were
`performed.
`complete
`blood
`lactate
`dehydrogenase
`with
`isoenzymes,
`complement,
`were
`checked
`before,
`during,
`and
`after
`acid
`levels
`parameters
`included
`complete
`blood
`Pharmacokinetic
`platelet
`counts,
`cell-bound
`MDX-
`and
`with
`differential
`1.
`serum
`CD15,
`and
`serum
`MDX-1
`cells,
`CD15-positive
`after
`studied
`before,
`during,
`and
`were
`parameters
`1 infusion.
`for
`marrow
`aspirates
`were
`obtained
`Bone
`patient
`cytometry
`assays
`were
`obtained
`at
`and
`flow
`end
`of
`the mAb
`infusion
`(24
`h). Complete
`and
`at
`the
`performed
`weekly
`for
`1 2 weeks
`after
`infu-
`counts
`were
`Physical
`examination,
`performance
`status,
`serum
`chemis-
`serum
`electrolytes,
`routine
`urinalysis,
`and
`PTIPTT
`were
`4,
`at
`8,
`and
`12 weeks
`after MDX-1
`infusion.
`HAMA
`I
`were
`done
`at 2, 4, 8,
`and
`12 weeks
`after
`infusion.
`Toxicity
`Measurements.
`Toxicity
`during
`and
`
`infusion
`Cooperative
`
`was
`
`assessed
`Oncology
`
`and
`Group
`
`graded
`(ECOG)
`
`according
`toxicity
`
`the
`to
`grading
`
`Eastern
`scale.
`
`after
`
`the
`
`
`
`Downloaded from on August 31, 2015. © 1995 American Association for Cancerclincancerres.aacrjournals.org
`
`
`Research.
`
`PETITIONER'S EXHIBITS
`
`Exhibit 1010 Page 3 of 9
`
`

`
`968
`
`Phase
`
`I Clinical
`
`Trial
`
`of Serotherapy
`
`in Patients
`
`with AML
`
`Patient
`
`Age/sex
`
`Diagnosis
`
`Table
`
`1
`
`Patient
`
`characteristics
`
`Initial
`
`blood
`
`cell
`
`counts
`
`No. of
`relapses
`
`WBC
`1000)
`
`(X
`
`% Blast
`
`Hemoglobin
`
`P
`
`101
`102
`103
`104
`105
`106
`107
`108
`109
`110
`301
`302
`303
`304
`401
`402
`
`241M
`38/F
`671M
`62/M
`26/F
`56/M
`22/M
`61/M
`52/M
`60/F
`24fF
`37/M
`27/M
`46/M
`48/M
`67/M
`
`AML
`AML
`AML
`AML
`AML
`MDS(cid:1)
`AML
`MDS
`AML
`AML
`AML
`AML
`AML
`MDS
`AML
`AML
`
`a MDS, myelodysplastic
`
`syndrome.
`
`converted
`(secondary
`converted
`
`to AML
`to Hodgkin’s
`to AML
`
`disease)
`
`converted
`
`to AML
`
`1
`4
`2
`1
`2
`0
`0
`0
`1
`3
`4
`1
`1
`0
`3
`2
`
`27.2
`22.6
`1.3
`66
`1.7
`9.2
`21.9
`7.8
`16.3
`0.4
`9.4
`2.9
`10.6
`73.1
`6.9
`3.4
`
`21
`98
`34
`5
`4
`84
`75
`13
`24
`24
`83
`0
`0
`84
`36
`5
`
`9.5
`8.4
`9.9
`10.1
`8.7
`8.4
`9.3
`7.6
`8.6
`9.6
`12.1
`10.9
`12
`8.5
`9.9
`10.5
`
`RESULTS
`exception
`the
`With
`are
`based
`discussion
`and
`MDX-1
`1, during
`infusion
`the
`infusion
`(infusion
`end,
`in
`therapy.
`Thus,
`decreases
`effects
`results
`are
`due
`to
`the
`not
`the
`subsequent
`chemotherapy.
`
`results,
`of HAMA
`obtained
`on
`data
`1,
`of MDX-i
`prior
`24)
`=
`counts
`cell
`of MDX-1
`
`t
`
`results
`following
`the
`of
`before
`infusion
`after
`immediately
`and
`chemo-
`receiving
`to
`pharmacokinetics
`and
`1 immunotherapy
`
`and
`
`and
`fever
`of mild
`reactions
`adverse
`reported
`Two
`reactions.
`2.
`grade
`toxicity
`Institute
`Cancer
`National
`mild
`hypotension,
`re-
`and
`hypotension
`experienced
`109)
`One
`patient
`(patient
`short-
`pain,
`aching,
`back
`vomiting,
`ported
`dizziness,
`nausea,
`and
`was
`stopped,
`infusion
`The mAb
`ness
`of breath,
`fever.
`and
`with
`and
`infused
`for
`the
`symptoms
`the
`patient
`treated
`was
`the
`restarted
`after
`infusion
`was
`mAb
`normal
`The
`saline.
`(approximately
`45 mm).
`adverse
`The
`symptoms
`subsided
`to be mild
`(ECOG
`grade
`II or
`less).
`were
`events
`considered
`104)
`treated
`at
`1.0-mg/kg
`level
`eighth
`The
`patient
`(patient
`experienced
`an
`acute
`respiratory
`episode
`reported
`nau-
`fever.
`sea,
`vomiting,
`dizziness,
`and
`The
`was
`stopped
`after
`20 mm,
`and
`the
`symptoms
`resolved
`h
`after
`3
`treatment.
`Although
`the
`adverse
`events
`were
`II or
`less,
`not
`the
`infusion
`was
`restarted.
`receiving
`five
`Three
`the
`patients
`reported
`completed
`infusion.
`One
`patients
`lated
`to
`infusion.
`Two
`patients
`(ECOG)
`One
`these
`diaphoresis,
`enced
`pain,
`restarted.
`infusion
`not
`second
`solved
`The
`toxicities
`hypotension,
`the
`gestion,
`Two
`of
`IV (ECOG)
`fatigue,
`chills,
`tachycardia,
`and
`patient
`died
`This
`Gram-negative
`(patient
`pain,
`back
`the
`7 mm into
`patient
`subsequently
`intravascular
`distress.
`infusion
`
`in
`
`of
`
`ages
`Phase
`the
`
`Patients
`between
`patients
`Sixteen
`treated
`were
`47)
`years
`age,
`>80%,
`Karnofsky
`status
`positive
`blast
`cells
`>20%
`were
`pulmonary
`status
`results
`nary
`function
`test
`left
`ventricular
`function.
`teristics.
`
`and
`22
`the
`study.
`I
`this
`criteria
`all met
`and
`Exceptions
`for MDX-11.
`patients
`who
`made
`for
`two
`that were
`suboptimal
`or had
`Table
`1 lists
`individual
`patient
`
`(median
`67
`had
`Each
`for
`beukemic
`for
`cardio-
`had
`pulmo-
`decreased
`charac-
`
`the
`
`in
`
`h
`
`the
`
`a
`
`1.
`
`Toxicity
`of MDX-i
`dose
`intended
`received
`patients
`Thirteen
`experienced
`serious
`and
`401)
`303,
`104,
`(jatients
`patients
`Three
`infusion.
`Another
`pa-
`of
`the
`termination
`necessitating
`reactions
`reactions
`necessitating
`milder
`experienced
`tient
`(patient
`106)
`complete
`infusion
`taking
`the
`resulted
`infusion
`delays
`that
`Patient
`109
`also
`experi-
`h.
`24
`intended
`than
`33.5
`rather
`of
`to interuption
`the
`infusion
`leading
`problems
`enced
`minor
`were
`still
`included
`results
`patient’s
`45 mm. This
`in the
`pharma-
`Pharmacokinetic
`data
`for
`other
`patients
`cokinetic
`analyses.
`to insufficient
`numbers
`of cells
`for
`analysis.
`incomplete
`due
`majority
`toxicities
`were
`common
`side
`effects
`experienced
`of
`infusions
`or minor
`allergic
`reactions
`that
`resolved
`mAb
`during
`upon
`(Table
`2).
`treatment
`0.5-mg/kg
`the
`at
`patients
`treated
`All
`three
`experiencing
`the MDX-
`1 1 infusion
`without
`patients
`treated
`to
`the mAb.
`Of
`the
`eight
`level,
`four
`completed
`the MDX-1
`1 infusion
`
`for
`
`are
`The
`
`level
`adverse
`at
`the
`with
`
`tolerated
`reactions
`1.0 mg/kg
`no
`adverse
`
`of
`the
`the
`events.
`hypotension,
`was
`stopped
`upon
`treatment.
`experienced
`and
`sneezing.
`experienced
`105)
`
`of
`chest
`and
`
`grade
`
`mg/kg
`reported
`(patient
`distress,
`respiratory
`infusion.
`MDX-11
`underlying
`to
`an
`second
`The
`experienced
`breath,
`halted,
`fever,
`sis,
`upon
`
`patient
`severe
`nausea
`and
`the
`and
`disseminated
`and
`respiratory
`treatment.
`
`The
`
`not
`
`the
`and
`infusion
`within
`grade
`
`1.5 mg/kg
`adverse
`no
`experienced
`(patient
`and
`
`re-
`II
`
`MDX-il
`events
`grade
`experi-
`The
`re-
`II
`
`con-
`1.5
`at
`patient
`fever,
`the
`due
`co!i).
`event
`of
`was
`
`IV
`shortness
`infusion
`tachycardia,
`rhabdomyoly-
`all
`resolved
`
`401)
`nausea.
`symptoms
`The
`with
`grade
`patient
`chills,
`sinus
`fever,
`treated
`patients
`One
`events.
`hypotension,
`but
`completed
`4 days
`after
`infusion
`sepsis
`(Escherichia
`with
`a
`grade
`303)
`burning,
`facial
`infusion.
`The
`developed
`coagulation,
`These
`symptoms
`was
`restarted.
`
`
`
`Downloaded from on August 31, 2015. © 1995 American Association for Cancerclincancerres.aacrjournals.org
`
`
`Research.
`
`PETITIONER'S EXHIBITS
`
`Exhibit 1010 Page 4 of 9
`
`

`
`Clinical
`
`Cancer
`
`Research
`
`969
`
`Table
`
`2
`
`Toxicities
`
`e xperienced
`
`by
`
`patients
`
`dun
`
`ng
`
`PM-81
`
`infusion”
`
`Severity
`
`Relation
`
`to MDX-11
`
`Symptom
`
`Patient
`count
`
`Mild
`
`Moderate
`
`Severe
`
`Possible
`
`Probable
`
`Highly
`probable
`
`0
`0
`0
`0
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`1
`0
`0
`0
`0
`1
`1
`1
`1
`1
`1
`I
`1
`1
`1
`I
`0
`0
`
`0 0
`
`0
`0
`0
`0
`0
`1
`1
`0
`0
`0
`0
`0
`0
`0
`0
`1
`0
`0
`0
`1
`0
`1
`0
`0
`0
`0
`0
`0
`0
`0
`2
`0
`
`0 0
`
`1
`1
`1
`1
`2
`0
`0
`1
`1
`0
`0
`0
`0
`0
`0
`1
`2
`1
`1
`2
`2
`0
`1
`1
`0
`0
`0
`0
`0
`0
`1
`2
`
`1 1
`
`0
`0
`0
`0
`1
`0
`0
`0
`0
`1
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`1
`0
`1
`1
`1
`0
`1
`0
`
`0 1
`
`1
`1
`1
`0
`1
`1
`2
`2
`0
`0
`0
`0
`1
`1
`0
`0
`1
`1
`0
`4
`2
`2
`2
`2
`0
`1
`0
`0
`0
`1
`2
`2
`
`0 0
`
`0
`0
`0
`1
`1
`1
`0
`0
`2
`0
`1
`1
`0
`0
`1
`2
`1
`0
`1
`0
`1
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`1 0
`
`1/3
`1/3
`1/3
`1/3
`3/8
`2/8
`2/8
`2/8
`2/8
`1/8
`1/8
`1/8
`1/8
`1/8
`1/8
`2/8
`2/8
`1/8
`1/8
`4/5
`3/5
`2/5
`2/5
`2/5
`1/5
`1/5
`1/5
`1/5
`1/5
`1/5
`3/5
`2/5
`1/5
`1/5
`
`Headache
`Chest
`pain
`Nausea
`Rash
`Dyspnea
`Chills
`Hypotension
`Nausea
`Vomiting
`Bronchospasm
`Asthenia
`Headache
`Back
`pain
`Vasodilation
`Hypoxia
`Pruritus
`Fever
`Tachycardia
`Myalgia
`Fever
`Nausea
`pain
`Chest
`Tachycardia
`Dyspnea
`Back
`pain
`Vasodilation
`Hypoxia
`Bihirubinemia
`Myopathy
`Pruritus
`Hypotension
`Chills
`Asthenia
`Blood
`
`pressure
`
`instability
`
`Dose
`(mg/kg)
`
`0.5
`
`1.0
`
`1.5
`
`a Toxicities
`
`listed
`
`in order
`
`of
`
`dose
`
`and
`
`probable
`
`relation
`
`to MDX-1
`
`1.
`
`HAMAs
`
`developed
`patient
`
`This
`
`HAMAs
`was
`treated
`
`noted
`with
`
`after
`4 weeks
`the
`0.5-mg/kg
`
`Table
`
`3
`during
`
`Patient
`infusion,
`
`blast
`at
`
`(X
`
`1000)
`counts
`end,
`and
`infusion
`chemotherapy
`
`before
`after
`
`MDX-1
`infusion
`
`1 infusion,
`during
`
`Only
`infusion
`dose
`
`patient
`one
`60).
`(titer,
`1.
`of MDX-1
`HAMA
`assays
`HAMA
`IgG)
`for
`experienced
`severe
`were
`within
`the
`sera.
`The
`screening
`patient
`experiencing
`for
`elevated
`levels
`normal
`ranges.
`
`1gM and
`for
`performed
`were
`reactions.
`adverse
`range
`as
`normal
`and
`1-week
`sera
`the most
`severe
`IgE antibody
`
`of
`
`with
`three
`for
`
`to
`
`IgE (along
`on
`all
`litres
`compared
`postinfusion
`reaction
`but were
`
`all
`
`usual
`the
`patients
`three
`pooled
`sera
`
`assay
`who
`patients
`human
`from the
`also
`tested
`to be within
`
`were
`found
`
`Patient
`
`Infusion
`t
`
`start
`0
`
`t
`
`12 h
`
`Infusion
`t
`
`=
`
`end
`24 h
`
`Postchemotherapy
`start
`48 h
`
`t
`
`=
`
`2.09
`1.25
`0.14
`0.00
`ND”
`ND
`3.61
`0.00
`12.32
`0.00
`93.12
`0.00
`
`10.91
`8.46
`0.41
`0.01
`3.04
`0.00
`1.37
`0.00
`11.19
`0.00
`65.14
`0.09
`
`2.34
`33.25
`0.40
`0.02
`4.06
`1.17
`1.15
`0.00
`9.49
`0.00
`33.07
`0.06
`
`101
`102
`103
`105
`107
`108
`109
`110
`301
`302
`304
`402
`a ND,
`
`5.71
`22.15
`0.44
`0.07
`16.43
`1.01
`3.91
`0.10
`7.80
`0.00
`70.18
`0.00
`not done.
`
`Pharmacokinetics
`
`Serum
`with
`
`MDX-!!
`the
`initial
`
`Levels.
`reservoir
`
`dose
`The
`of CD15,
`
`1 received,
`of MDX-i
`whether
`cell-bound
`(on
`
`along
`
`Effect
`
`in
`
`and Blast Counts
`on WBC
`Infusion
`of
`total WBC
`had
`a decrease
`12
`patients
`of
`(75%)
`Nine
`Seven
`of
`the
`of
`the MDX-11
`infusion.
`the
`completion
`at
`count
`patients
`with
`decreased
`WBCs
`had
`a 50% or greater
`tran-
`nine
`decrease
`in their WBC.
`Blast
`counts
`at
`the
`infusion
`end
`sient
`decreased
`for
`8 (67%)
`of
`the
`12 patients,
`with
`as many
`patients
`experiencing
`a 50% or greater
`transient
`decrease
`in their
`blast
`counts
`(Table
`3). Significant
`transient
`decreases
`of WBCs
`and
`blast
`counts
`were
`seen
`at
`three MDX-i
`1 doses.
`
`all
`
`
`
`Downloaded from on August 31, 2015. © 1995 American Association for Cancerclincancerres.aacrjournals.org
`
`
`Research.
`
`PETITIONER'S EXHIBITS
`
`Exhibit 1010 Page 5 of 9
`
`

`
`970
`
`Phase
`
`I Clinical
`
`Trial
`
`of Serotherapy
`
`in Patients
`
`with AML
`
`Table
`
`4
`
`PM-81
`
`dose
`
`vs. maximum
`
`serum PM-81
`
`levels
`
`achieved
`
`during
`
`infusion
`
`and
`
`initial
`
`(t
`
`=
`
`0) cell
`
`counts
`
`(X
`
`1000/mI)
`
`Dosage
`(mg/kg)
`
`Maximum
`PM-81
`
`serum
`(p.gJml)
`
`1.5
`1.5
`1.0
`1.0
`1.0
`1.0
`1.0
`1.0
`1.0
`0.5
`0.5
`0.5
`
`3.56
`1.46
`4.76
`1.06
`1.02
`0.75
`<0.50
`<0.50
`<0.50
`0.00
`0.00
`0.00
`
`Patient
`
`105
`107
`110
`402
`301
`302
`108
`109
`304
`103
`102
`101
`
`a ND,
`
`not
`
`done.
`
`Initial
`WBC
`
`1.7
`21.9
`0.4
`3.4
`9.4
`2.9
`7.8
`16.3
`73.1
`1.3
`22.6
`27.2
`
`Initial
`blast
`count
`
`% CD15-
`positive
`blasts
`
`Initial
`positive
`
`CD15-
`blasts
`
`0.1
`16.4
`0.1
`0.0
`7.8
`0.0
`1.0
`3.9
`70.2
`0.4
`22.2
`5.7
`
`93
`100
`64
`64
`29
`99
`89
`82
`ND”
`87
`100
`45
`
`0.1
`16.4
`0.1
`0.0
`2.3
`0.0
`0.9
`3.2
`ND
`0.4
`22.2
`2.6
`
`Table
`
`5
`
`Serum CD15
`
`and serum PM-81
`
`levels
`
`vs. percentage
`
`of
`
`leukemic
`
`bone
`
`marro w blasts
`
`with
`
`cell-bound
`
`PM -81
`
`at
`
`infusion
`
`end
`
`(t
`
`=
`
`24)”
`
`Serum PM-81
`(p.g/ml)
`
`Maximum
`PM-81
`
`serum
`
`bone marrow
`24-h
`% Pos.
`PM-8l
`
`65
`64
`60
`16
`ND
`ND
`16
`11
`ND
`ND
`
`0 0
`
`3.56
`1.46
`4.76
`1.06
`1.02
`0.75
`<0.50
`<0.50
`<0.50
`0.00
`0.00
`0.00
`
`Serum CD15
`(mg/mh)
`
`Patient
`
`t
`
`0
`
`t
`
`24
`
`t
`
`0
`
`t
`
`24
`
`105
`107
`110
`402
`301
`302
`108
`109
`304
`101
`102
`103
`
`0.24
`0.59
`0.13
`0.38
`0.71
`0.59
`0.99
`1.26
`0.10
`0.59
`0.30
`0.27
`
`0.00
`0.00
`0.00
`0.00
`0.00
`0.00
`0.00
`0.12
`0.00
`1.24
`0.00
`0.00
`
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`0
`
`2.10
`1.46
`4.76
`1.06
`1.02
`0.00
`<0.50
`0.00
`<0.50
`0.00
`0.00
`0.00
`
`a p00(cid:1)j
`b ND,
`
`human
`not done.
`
`sera
`
`control
`
`averaged
`
`0.32 mg/ml
`
`serum
`
`CDI5.
`
`or
`
`levels
`serum
`the
`affected
`soluble,
`1 (0.5
`of MDX-1
`lowest
`dose
`the
`receiv-
`all patients
`mAb,
`while
`MDX-
`had
`quantifiable
`serum
`detect-
`dose
`(1.0 mg/kg)
`had
`1 levels
`4).
`Serum
`MDX-1
`h and
`returned
`to 0 within
`
`24
`
`or
`cells)
`leukemic
`normal
`receiving
`1 . Patients
`of MDX-i
`no
`detectable
`serum
`had
`mg/kg)
`highest
`dose
`(1.5 mg/kg)
`ing
`the
`1 1 . Patients
`receiving
`the median
`able
`serum
`MDX-!
`1 levels
`(Table
`typically
`peaked
`between
`12 and
`24
`h after
`the
`infusion.
`in
`The
`amount
`of MDX-1
`higher
`in patients
`who
`received
`highest
`CD15
`initial
`reservoir
`cell-bound
`lower
`of
`patients
`or
`tumor
`burden).
`Conversely,
`higher
`the
`doses
`of
`antibody,
`and/or
`1 present
`had
`CD15,
`little
`to
`no MDX-1
`there was
`a high
`percentage
`of blasts
`with
`4).
`as
`in patient
`301
`(Table
`were
`Soluble
`CD1S
`levels
`MDX-1
`1 levels;
`patients
`in general,
`their
`serum
`CD15
`0 for
`at
`t
`=
`serum
`levels
`of MDX-1
`1 [with
`whose
`initial
`cell-bound
`CD15
`significantly
`higher
`than
`the
`other
`expected,
`patients
`had
`either
`free
`
`I
`
`present
`the
`
`had
`
`sera
`doses
`(lower
`who
`reservoirs
`in their
`low CD15
`
`be
`to
`tended
`the
`had
`and
`initial WBC
`received
`lower
`of cell-bound
`serum,
`unless
`expression,
`
`to
`related
`also
`lowest
`the
`with
`achieved
`dosage
`the
`exception
`(WBC
`and
`patients;
`antibody
`
`the
`
`of
`
`serum
`levels
`highest
`304
`were
`5]. As
`antigen
`
`the
`patient
`counts)
`4 and
`free
`
`of
`
`blast
`Tables
`or
`
`t
`
`=
`
`find
`
`=
`
`not
`
`or
`
`1,
`
`=
`
`1
`
`0 and
`at
`serum
`in their
`present
`did we
`case
`no
`in
`24;
`t
`=
`simultaneously.
`present
`CD1S
`serum
`1 and
`serum
`MDX-i
`both
`patients
`to have
`serum
`CD15
`the only
`109 were
`101
`and
`Patients
`5). Note
`that
`patient
`101
`received
`still
`present
`at
`24
`(Table
`t
`had
`the
`second
`highest
`initial
`of MDX-11,
`the
`lowest
`dose
`achieved
`detectable
`levels
`of
`serum MDX-1
`WBC,
`and never
`109
`had
`a very
`high
`initial
`serum
`CD1S
`level
`whereas
`patient
`have
`detectable
`serum
`MDX-1
`1. Thus,
`the
`and
`also
`did
`(on
`of CD15,
`whether
`cell-bound
`normal
`or
`initial
`reservoir
`was
`soluble,
`important
`in determining
`a patient’s
`leukemic
`cells)
`levels
`serum
`of MDX-il.
`Serum
`levels
`of CD15
`to achieve
`ability
`patient’s
`to the
`initial WBC
`or blasts
`counts.
`were
`unrelated
`MDX-!!.
`Cell-bound
`MDX-1
`1 on circulat-
`Cell-bound
`followed
`throughout
`the
`duration
`the
`infusion,
`blasts
`was
`of
`ing
`a bone marrow
`biopsy
`was
`obtained
`at
`the
`infusion
`end
`and
`(t
`of
`h).
`Serum
`levels
`of MDX-1
`i
`correlated
`with
`the
`amount
`24
`MDX-i
`1 reaching
`the
`leukemic
`blasts
`in marrow
`(Table
`5).
`had
`Those
`patients
`who
`had
`no detectable
`serum
`MDX-1
`1 levels
`those
`no detectable
`cell-bound
`MDX-1
`i
`in their marrows,
`while
`patients
`with
`serum
`levels
`of MDX-1
`1 had
`cell-bound
`MDX-!
`on
`their
`marrow
`blasts.
`The
`percentage
`MDX-1
`1 was
`
`bound
`
`of
`also
`
`peripheral
`an indicator
`
`blasts
`leukemic
`of
`cell-bound
`
`with
`MDX-1
`
`cell-
`1 in
`
`
`
`Downloaded from on August 31, 2015. © 1995 American Association for Cancerclincancerres.aacrjournals.org
`
`
`Research.
`
`PETITIONER'S EXHIBITS
`
`Exhibit 1010 Page 6 of 9
`
`

`
`leukemic
`marrow
`able
`serum
`levels
`cell-bound
`MDX-1
`with
`cell-bound
`circulating
`blasts
`
`patients
`Those
`blasts.
`of MDX-1
`1 and
`>50%
`1 had
`(cid:18)60%
`leukemic
`MDX-1
`1. Patients
`with
`with
`cell-bound
`MDX-i
`
`both
`
`quantifi-
`had
`who
`blasts
`with
`circulating
`blasts
`in their marrow
`lower
`percentages
`1 (patients
`108
`
`of
`and
`
`or
`
`could
`
`be
`quantified
`not
`of MDX-i
`levels
`serum
`1
`that
`percentage
`lower
`had
`and
`108
`109)
`a significantly
`marrow
`I(cid:1)Si6%).
`their
`MDX-1
`i-positive
`blasts
`in
`blasts
`typically
`MDX-1
`1 on
`circulating
`leukemic
`between
`4 and
`12
`h and
`returned
`to normal
`24
`h after
`
`of
`
`402)
`(patients
`cell-bound
`Cell-bound
`peaked
`infusion.
`percentage
`CD15-positive
`throughout
`cells
`did
`not
`vary
`quantifiable
`sion.
`Those
`patients
`of CD1S-positive
`still
`had
`high
`percentages
`antigen
`on
`the
`cell
`presence
`of unbound
`indicated
`that
`the
`of CD15-positive
`cells
`with
`antibody
`fluorescent
`intensity
`cells
`was
`also
`affected
`serum
`levels
`
`The
`
`cells.
`significantly
`with
`
`(the
`not
`
`by
`
`infu-
`levels
`the
`
`of CD15-positive
`the MDX-il
`MDX-11
`serum
`indicating
`cells,
`Measu