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`MAUR VIAVAINKTIVIE SID, BRE SENNA SWAM COME%
`
`UNITED STATES DEPART.\IE~T OF COMMERCE
`UNITED STATES DEPARTMENT OF COMMERCE
`United States Patent and Trademark Office
`United States Patent and Trademark Office
`
`March 14, 2022
`March 14, 2022
`
`THIS IS TO CERTIFY THAT A~TEXED HERETO IS A TRUE COPY FROi\1
`THIS IS TO CERTIFY THAT ANNEXED HERETO IS A TRUE COPY FROM
`THE RECORDS OF THIS OFFICE OF:
`THE RECORDS OF THIS OFFICE OF:
`
`Pre-Grant Publication Number: 2004/0126363
`Pre-Grant Publication Number: 2004/0126363
`Pre-Gr ant Publication Date: July 1, 200.:f.
`Pre-Grant Publication Date: July 1, 200-1
`
`By Authority of the
`By Authority of the
`Under Secretary of Commerce for Intellec al Property
`Under Secretary of Commerce for Intellec al Property
`and Director of the United States Patent d T ·ademark Office
`and Director of the United States Patent d T •ademark Office
`
`• /4
`C rt .'ing Office1· tJ
`
`y la o~
`is
`olley
`yl
`ing Officer
`rt
`C
`
`C
`
`_
`
`Miltenyi Ex. 1018 Page 1
`
`

`

`iu uu mi
`
`
`111111111 1111111111
`1111111111111111 IIIIII IIIII 11111 1111111111 lllll lllll lllll 111111111111111 1111111111 111111111
`US 20040126363A I
`
`(19) Uni1ted States
`(19) United States
`c12> Patent Application Publication
`(12) Patent Application Publication
`Jensc10 et al.
`Jensen et al.
`
`( JU) Pub. No.: US 2004/0126363 Al
`(10) Pub. No.: US 2004/0126363 Al
`Jul. 1, 2004
`Jul. 1, 2004
`(4:1) Pub. Date:
`(43) Pub. Date:
`
`(54) C l>19-SPl~CfFJC REDIIU◄:CTlm IMMUNI<:
`(54) CD19-SPECIEIC REDIRECTED IMMUNE
`CELLS
`CELLS
`
`(76)
`(76)
`
`lnventors: Miclmcl C . .Jensen, Pa~adena. CA
`Inventors: Michael C. Jensen, Pasadena, CA
`(US); Stephen Forman, San Marino,
`(US); Stephen Forman, San Marino,
`CA (US); Andrew Raubitschek, San
`CA (US); Andrew Raubitschek, San
`Marino, CA (US)
`Marino, CA (US)
`
`Correspondence /\ddrcf<.-;:
`Correspondence Address:
`ROTt-llWf+:LI,, FIGG, ERNST & MANHECK,
`ROTHWELL, FIGG, ERNST & MANBECK,
`P.C.
`P.C.
`1425 K STREE'J~ N. W.
`1425 K STREET, N.W.
`SUITE 800
`SUITE 800
`WAS.HINGTON, De 20005 (US)
`WASHINGTON, DC 20005 (US)
`
`(21) Appl. No.:
`(21)
`Appl. No.:
`
`10/416,011
`l0/416,01 I
`
`(22) PCT Filed:
`PCT Filed:
`(22)
`
`Nov. 7, 2001
`Nov. 7, 2001
`
`(86) PCl' Nlo.:
`(86)
`PCT No.:
`
`PCT/US0J/42997
`PCT/US01/42997
`
`Related U.S. Applicntion Data
`Related U.S. Application Data
`
`(6())
`Provisional application No. 60/246,117, tiled on Nov.
`(60) Provisiional ~pplication No. 60/24G, I I 7, lilcd on Nov.
`7,2000.
`7, 2000.
`
`J'ublication Classification
`Publication Classification
`
`(51) Int.
`(51)
`lnt. C t.7 ............................. A61K 48/00; Cl2N 5/08
`
`A61K 48/00; C12N 5/08
`(52) U.S. Cl .
`...................... 424/93.21; 424/93.7; 43.5/372
`(52) U.S. Cl.
`424/93.21; 424/93.7; 435/372
`
`(57)
`(57)
`
`ABSTRACT
`ABSTRACT
`
`Genetically engineered, CDl 9-spcdfic redirected [rnmuoe
`Genetically engineered, CD19-specific redirected immune
`cells expressing s cell surface protcio baving an extracdlu lar
`celLs expressing a cell surface protein having an extracellular
`domain comprising a receptor wbieh i.s specific for CD l 9. au
`domain comprising a receptor which is specific for CD 19, an
`iotraudlular signaling domain, am.I
`intracellular signaling domain, and a
`transmembrane
`transmernbrane
`II
`domain. Use of sucb cells for cellu lar immunotherapy of
`domain. Usc of such cells for cellular immunotherapy of
`CDJ 9+ malignaacies aad f0r abrogating any untoward B cell
`CD19+ malignancies and for abrogating any untoward B cell
`ruoctioo. To one embuclinu:nt, th.: immuo..: c.:ll is a Tcell and
`function. In one embodiment, the immune cell is a Tcell and
`the cell surface protei n is a single chain scfvFc:t rec:eptor
`the cell surface protein is a single chain scFvFc:t receptor
`where scFv designates the VH and V t. chains of a i;ingle
`where scFv designates the Vn and V,. chains of a single
`cbain monoclonal antibody to CD I 9, Fe represents al least
`chain monoclonal antibody to CD19, Fc represents at least
`par! of a coaslaol region of ao lgG,, and t rcprcscnL-; tbc
`part of a constant region of an IgG,, and
`represents the
`ia1racellular signaling domain of lho zcia chain of bumao
`intracellular signaling domain of the zeta chain of human
`CD3. The cxtracellular domain scFvFc and the intracellular
`CD3. Tbe extracellular domain sc¥v11c and th.: intrac~,Uular
`domain t arc linked by a lransmcmbrane domain such ~s tbll
`domain t are linked by a transmembrane domain such as the
`lransmembrane domain of CD4. A metbocl of ma.king a
`transmembrane domain of CD4. A method of making a
`redirected T cell expressing n chimeric T cell receptor by
`redirected T cell expressing a chimeric T cell receptor by
`lllecLroporation u.sing naked DNA encoding Lhe receptor.
`electroporation using naked DNA encoding the receptor.
`
`Miltenyi Ex. 1018 Page 2
`
`

`

`Patent Application Publication
`Patent Application Publication
`
`Jul. 1, 2004 Sheet 1 of 8
`Jul. 1, 2004 Sheet 1 of K
`
`US 2004/0126363 Al
`US 2004/0126363 Al
`
`Xba!
`XbaI
`
`(l'JGMCSF s; .:;na l ;:ieptide-)
`1 peptide-)
`• (hGMCSF s:gna
`s
`::
`L L L C E
`P
`L
`7 7 S
`!..
`i..
`'.'
`-:-
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`L
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`L
`L
`£
`M
`L
`~
`ATCTCTAGAG CCGCCACCAT GCTTCTCCTG GiGACAAGCC TTCTGCTC".'G ,GAGTrAC::CA
`G7GACAAGCC
`TTCTGCTCTG TGAGTTACCA
`1 ATCTCTAGAG CCGCCACCAT GCTTCTCCTG
`TAGAGATCTC GGCGGTGGTA CGAAGAGGAC CACfG17GGG AAGACGAGAC AC':-CAATGGT
`AAGACGAGAC AC7CAATGGT
`CACTGTTCGG
`TAGAGATCTC GGCGGTGGTA CGAAGAGGAC
`
`(V~ - l
`Q 7 7 S S L S
`0:-1~ Q 7 7 5 5~3
`?
`::>
`-
`F L ~
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`Q M
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`:l CACCCAGCAT TCCTCC~GAT
`CAGATGACAC AGACTACA~C C1CCCTG7C!
`CCCAGACATC
`AGACTACATC CTCCC7GiCT
`CAGATGACAC
`TCCTCCTGA7 CCCAGACATC
`61 CACCCAGCAT
`GTCTACTGTG TCTGA:~TAG G~GGGACAGA
`GGGTCTGTAG
`G:GGGICGTA AGGAGGACTA
`7CTGA7-TAG GAGGGACAGA
`GTCTACTGTG
`AGGAGGACTA GGGTCTGTAG
`G7GGGTCGTA
`s C
`., R A
`s
`T
`S X 'I
`1
`S Q
`rt
`R A
`A S
`G 0
`: S
`7
`D R V
`A S
`G
`i21 Gcc::'!'~~GG G_-,3ACAGAG7 ~ACCAIC;G: 7~=AGGGCM ~T:A~·~M:;~.-r -:.;G:;.,;....:.:-:.:..:(cid:173)
`GL:ACAGAG7 CACCATC;G:
`7-:=AGGG.Art
`121 GCC7CT'TGG
`CGGAuAi:iAC-'.: CTCTG7C':'CA :;TGGTAGTCA P.CGTCC':GTT CAGT::C7G7A A,CA:"!'TA,A.
`AiCAT7TATA
`ACGTCC7CTT
`CTCTG7C7CA GTGGTAGTCA
`CGGALAGACC
`
`L N W Y Q Q K
`H ~ S
`L L ~ Y
`P D G T V K
`L L I Y
`H I 5
`T V K
`L N W
`P D G
`Y Q Q K
`1B l TTM-.ATTGGT ATCAGGAGAA ACCAGATGGA ACTGTTAAAG TCCTGATCTJI CCJI .,.\CATCA
`CCA'\CATCA
`181 TTAAATTGGT
`ACTGTTAAAC
`TCCTGATCTP
`ATCAGCAGAA ACCAGATGGA
`AATTTAACCA TAGTCGTCTT TGGTCTACCT TGACAATTTG AGGAGTAGAT GGTATGTAGT
`AGGACTAGAT
`GGTATGTAGT
`AATTTAACCA
`TAGTCGTCTT TGGTCTACCT
`TGACAATTTG
`s G s G s G T
`s R F
`s G V
`p
`R L H
`R L H
`S G V
`G S G T
`D Y S
`S R F
`P
`S G S
`D
`5
`'{
`20 AGATTACACT CAGGAGTCCC ATCAAGGTTC' ?.GI'GGCAGTG GGTCTGGAAC AGATTATTCT
`241 AGATTACACT
`AGATTATTCT
`CAGGAGTCCC
`GGTCTGGAAC
`ATCAAGGTTC
`AGIGGCAGTG
`TCTAATGTGA GTCCTCAGGG TAGTTCCAAG TCACCGTCAC CCAGACCTTG TCTAATAAGA
`TCTAATGTGA
`CCAGACCTTG
`TCTAATAAGA
`GTCCTCAGGG
`TCACCGTCAC
`TAGTTCCAAG
`
`L T
`s N L E
`L T I
`y
`I
`Q E D
`S N L E
`Y F C Q
`• G N
`1 A T
`I A T
`Q E
`0
`3Cll C':'::ACCATTA GCAACCTGGA GCAAGAAGAT ATTGCCACTT ACTTTTGCCA ACAGGGTAA'!
`301 C7TACCATTA
`GCAACCTGGA
`ACAGGGTAAT
`GCAAGAAGAT
`ACTTTTGCCA
`ATTGCCACTT
`GAGTGGTAAT CGTTGGACCT GGTTCTTCTA ,AACGGTGAA TGAAAACGGT TGTCCCATTA
`GAGTGGTAAT
`CGTTGGACCT
`CGTTCTTCTA
`TGTCCCATTA
`TGAAAACGGT
`TAACGGTGAA
`
`r C Q " G N
`
`,Wr.itlow linker- )
`,Whitlow linker-
`. G G G
`Y r
`I T G 5
`T S G
`K
`
`T -
`E
`T L
`T L ? Y
`G S T S G
`F G
`T
`G G T
`K L E
`T
`I
`?
`361 ACGCT~CCGT ACACGTTCGG AGGGGGGACT AAGT;GGAAA TAACAGGCTC CACCTCTGGA
`361 ACGCTTCCGT ACACGTTCGG AGGGGGGACT
`TAACAGGCTC CACCTCTGGA
`AAGTTGGAAA
`;GCGAAGGCA TG,GCAAGC: rc::ccccTGA ~-=:AACCTTT,ATTGTCCGAG GTGGAGACC7
`TGCGAAGGCA TGTGCAAGCC .‘
`.ATTGTCCGAG GTGGAGACCT
`TGA
`777AACCTTT
`:·,~- :
`:;
`s
`:;
`:<
`::;
`S K
`F 3 S 3
`C
`-
`K
`• S S
`7 K G
`E
`K
`~ ~ ~ E
`-
`::; S
`~
`:'CCGGCA,;]:
`42:C
`:GAGGG;..rcc- Ac:.:..AGGGCG AG3TGAAACT GCAGGAGTCr.
`421 7CCGGCAA::
`GCAGGAGTCA
`CCSGATC7GG
`AGGTGAAACT
`CGACGGATCC
`AC7AAGGGCG
`,l.~Gcc::;:-:-::G
`Gc:::c::ASG :GGT':'CCCGC TCCACTTTGA CGTCCTCAGT
`. _
`_
`TCCACTTTGA
`CGTCCTCAGT
`G,.-C7.AGG
`....
`..,
`s
`E' G
`;,
`s V
`s
`T C
`G
`s G
`G F G
`!.,
`S
`L S V
`T C T V
`• G V
`'! V
`'I
`GGACCTGGCC
`4 BJ,
`CTCAC,t,GAGC CTG7CCG7'C.A CA7G~ACTG7 -::TCAGGGG!'C
`TGG!GGCGCC
`481 GGACCTGGCC
`TGG:GGCGCC
`CATGCACTG7
`CTCAGGGGTC
`CTCACAGAGC
`CTGTCCGTCA
`ccr:;GACCGG
`ACCACCGCGG
`GAGTG7CTCG GACAGGCAGT GTA.CGTGACA GAGTCCCCAG
`CCTSGACCGG
`GTACGTGACA
`GAGTCCCCAG
`ACCACCGCGG
`GAGTGTCTCG
`GACAGGCAGT
`.
`-
`s L
`s
`::. E w
`y ~ ' /
`:-;
`F.. K G
`S L P
`C
`?
`C Y . 7
`p
`S W :
`K G
`F
`• E W
`<'
`3 Q P
`P
`?. Q
`TCATr;.. . ;.:G
`541
`AAGCTGGATT CGCO.GCCT:: CACG/1.AAGGG TC':'GGAGTGG
`-~C7AT~::.;-:'G":'
`541 TCATTA.:CG
`4CTATSGTG7
`CACGAAAGGG
`TCTGGAGTGG
`AAGCTGGATT
`CGCCP.GCCTC
`AGTAATGGGC
`T'!'CGF>':C':'AA SCGG7CGGAG G7GC'!'TTCCC AGACCTCACC
`!GATAcc;..:;..
`AGTAATGGGC
`AGACCTCACC
`TGATACCACA
`GTGCTTTCCC
`TTCCACCTAA
`GCGGTCGGAG
`
`~
`
`L G
`·;
`.:;
`;;
`.
`s A
`·,t N
`.
`L G ;
`:
`E
`G S
`W
`5
`E 7 7
`N
`•
`K
`S A
`S R L
`·.J
`5 R L
`K
`CTGGGAG':'AA
`TATGGGG':'A:;
`601
`J'GAAACCACA :'ACTATAATT CAGCTCTCAA ATCCAGACTG
`601 CTGGGAGTAA
`TATGGGG7AS
`TGAAACCACA
`CAGCTCTCAA
`ATCCAGACTG
`TACTATAATT
`GACCCT:ATT
`ATAcccc;;:-=::
`ACTTTGGTGT .ll,TGA:'ATTAA GTCGAGAGTT TAGGTCTGAC
`GACCCTCATT
`ATACCCCA7C
`TAGGTCTGAC
`ACTTTSGTGT
`ATGATATTAA
`GTCGAGAGTT
`▪
`Q T
`~ M ~ S
`~ ~ X 5
`7
`~ S Q
`• I :
`\
`K
`K S Q
`N S
`~
`F
`M
`K
`• 0 T
`r
`661 ACCATCA1-A AGGACi....AC:'= ~AAGAGc~·-
`- -rTT~T':'AA AAATGAACAG TC~GCAAACT
`661 ACCATCA1-A
`AGGAZAAC7:
`:AAGAGC""
`--7TTCTTAA
`AAATGAACAG
`TCTGCAAACT
`TGGTAGTAGT TCCTGTTGAG GTTCTCGG, T ~AAAAGAATT TTTACTTGTC AGACGTTTGA
`TGGTAGTAGT
`TCC7G7TGAG
`,AAAAGAATT
`GTTCTCGG.T
`TTTACTTGTC
`AGACGTTTGA
`-
`• '
`y A ~
`~ G G s
`~ A K
`A
`-
`y
`y
`~ ~ -
`0
`A :
`Y
`• A K
`G G S
`~ V
`Y
`Y A M
`V
`721 GATGACACAS C~ATTTACTA C:GTGCCAAA CA~TAfTACT ACGGTGGTAG CTATGCTATG
`121 GATGACACAG
`C:ATTTACTA
`CA7TATTACT
`C7GTGCCAAA
`ACGGT3GTAG
`CTATGCTATG
`CTAC~GTGTC GGTA.nJI.TGA! GACACGGTTT ~,AA,AATGA TGCCACCATC GATACGATAC
`CTACTGTGTC
`GGTAAATGAT
`STAATAATGA
`GACACGGTTT
`TGCCACCATC
`GATACGATAC
`
`FIGURE' 1A
`fIGURE. 1 lA
`
`Miltenyi Ex. 1018 Page 3
`
`

`

`Patent Application Publication Jul. 1, 2004 Sheet 2 of 8
`Patent Application Publication
`Jul. 1, 2004 Sheet 2 of 8
`
`US 2004/012636~1 Al
`US 2004/0126363 A 1
`
`.:::::ger,- .
`3▪
`~ S
`3 V
`D Y W G Q ~
`K
`IE
`F
`S V 7
`3
`5
`D Y W 5
`7 ;..GAA:::::A.z.
`7 8 'i GAC' AC-:'GGG ::;rcAAGG;.,;: :7CAGTCAC.::: :;: =::::c:-CAG
`TAGAACC2AA
`57,77CC7CAG
`:TCAGTCACC
`781 GAC7AC7GGG GTCAAGGI-A:
`.=-.!C7!G~~TT
`CTGATGACCC :,;::;::~:::~ GAGTCAG7,G :AGAGGAG:C
`AITTTGGGT7
`:AGAGGAGTC
` 0AGICAGTG
`CTGATGACCC
`
`5 S
`5 5
`
`:::;
`D
`
`AGAAGAC7G
`
`K 7 H 7 C P
`341 AAAACTCACA
`77775AS-237 GCAC.:‘,.J.'.:0
`
`?
`?
`_,,,.. ·---- _,.
`C ?
`C:7GAACTZ.
`G7GCCCAGCA
`- ..;J\:~~<rJ:"".--
`CC7GG
`GGAC7TGAG5
`CACGGG7CG7
`
`'i
`::
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`7AGTCAG;..AG
`
`:<
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`.-... ..:.-.; ....... ---
`:(
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`-----,..,,.,. .... -
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`... ..,.,..,op,_,...,..
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`:;..1-_:...t..::c;_~_
`3-, ZAZA72:7
`=GAZAr.:C._
`..:....;r. ... r1- - - - -
`GAGAAGGGGG GT':'T'!'G-GGT': CCTGTGGGAG :,;::-AGAGGG :c:GGG.j,\C, .:cAGTG,.:..:.::
`CCAGTGEACG
`CCTGGGGACT
`7AZTAGAGGG
`GT7TTGGGT7 CCTGTGGGAG
`GAGAAGGG3G
`
`p
`
`.. -
`
`F
`
`- N ~ Y V D G
`_ ~ K
`E D P
`~ V S ~
`V V V
`V D G
`N W
`F
`Y
`3 V V
`I J K
`D V
`H
`S
`E D P
`96ll GTGGTGGTGG ACGTGAGCCA CGAAGACCCT GAGGTCAAGT TCAACTGGTA ~GTGGACGGC
`^GTGGACGGC
`TCAACTGGTA
`3AGGTCAAGT
`961
`ACGTGAGCCA
`CGAAGACCCT
`GTGGTGGTGG
`CACCACCACC TGCACTCGGT GCTTCTGGGA CTCCAGTTG-. AGTTGACCAT GCACCTGCCG
`GCACCTGCCG
`AGTTGACCAT
`CTCCAGTTCJ
`GCTTCTGGGA
`CACCACCACC
`TGCACTCGGT
`Q Y N s·- T
`'I R
`H N A K T K P
`- T Y
`V E V
`R
`R E E
`3 E V
`Q Y N S *
`R E E
`H N
`T K P
`K
`A
`CGGGAGGAGC AGTACAACAG CACGTACCGT
`1021. GTGGAGGTGC ATAATGCCAA GACAAAGCCG
`CACGTACCGT
`AGTACAACAG
`CGGGAGGAGC
`GACAAAGCCG
`1021
`GTGGAGGTGC
`ATAATGCCAA
`GCCCTCCTCG TCATGTTGTC GTGCATGGCA
`CACCTCCACG TATTACGGTT CTGTTTCGGC
`3TGCATGGCA
`TCATGTTGTC
`GCCCTCCTCG
`TATTACGGTT
`CTGTTTCGGC
`CACCTCCACG
`V S V L T V L H O O W ~ N
`::; K E Y K C
`Y K C
`E
`3 L T
`K
`G
`• V S
`N
`D W L
`V
`L H 0
`lOBJ. GTGGTCAGCG TCCTCACCGT CCTGCACCAG GACTGGCTGA ATGGCAAGGA ~TACAAGTGC
`GTACAAGTGC
`ATGGCAAGGA
`1081
`GTGGTCAGCG
`3ACTGGCTGA
`TCCTCACCGT
`CCTGCACCAG
`CACCAGTCGC AGGAGTGGCA GGACGTGGTC C:GACCGAC! TACCG!TCCT CATGT!CACG
`CATGTTCACG
`TACCGTTCCT
`CTGACCGACT
`AGGAGTGGCA
`CACCAGTCGC
`GGACGTGGTC
`
`~SK ;:..!(G
`A K G
`-K
`7 : 5 K
`!<VS t l i<AL PAP
`K V 5
`N K A L
`P A P
`ll~i AAGGTC:CCA ACAA.~Gcc:r CCCAG:::cc::c ~r:~A~AA.~; :cA:CTCC~. ~GCCNt~G~G
`AGCCAAAG5G
`:CA7CTCCAA
`1141 AAGGiC7CCA
`ACAAAGCC:T
`r.T.:SAGAAAA
`!TC:AGAGG! ~G7TT:~GGA GGG::~GCGG ~AG=~c~~: ~ ;~:AGAG5!7 TCGGr:~:c=
`TCG37:7CC:
`5G7AGAGGIT
`7G7T7CGGGA
`7AGC7C7777
`T7CCAGAGGT
`GGG7CGGGGG
`
`C
`?.
`E
`R
`•
`S
`;,20: CAG:C:.:-G.:i.:; ---------(cid:173)
`120:
`CAGCC:::GAZ
`AACCACAS27
`G7ACACL:75
`.. ..;..::: . ..; .. - __ -:;
`-r. - ..; . ..; ..:J\.:;-, ..,.
`
`F ? s
`--- ---· __ ,,..
`- . - .. ·""'""'- .
`
`--------•;,,,
`::;
`: A
`K 5
`•
`- . -
`... - -· . .
`.. -.
`.... -. --· · --
`..
`------- ---
`' .
`.." .
`. . . ..
`C3C757AGCG
`AAGA7AGGG7
`AC:GGA:ZGA :CAG777CC5
`.,
`::;
`3 1 ? E
`E S
`N
`',j
`N N •
`E S N
`~
`~
`v
`GAACAAC':' ->.::
`1321 GAGAGC.l>,A'i':: ~GCrlGC:G:.:;A
`1321
`GAGAGCAATZ
`GAACAAC7AC
`C'rTG~':'GA~:;
`CCG7:55CC7
`CTCTCGTTAC
`CTTG7772A.2
`,_....,lo,!.~~.;,\,. ·- .
`
`---· -.-. ..; _ __ :-____
`1_.
`
`-----------
`
`7 K N
`R
`----,-------
`GA:CAAGAA:
`5A5A:SAGC7
`-· -'~• · -- - .:
`- - - . . -:.--_ .. - ...:n
`::
`
`V
`
`._
`
`•
`
`_
`
`-_._,--
`
`•
`•.
`.
`.
`GCACCTCA::
`
`CTCTCG':'TAC ,_,..,,.-.-,,.. ---....
`J
`5
`~---~~T--- --,-.-----~-
`F
`5 5
`F L
`K
`':AAG:::;,.:::
`139:
`_._,_ ___ ,M_r . .:,
`~J"-•---!...,..
`7.7:7:7A=A5
`GTTCGAr,7GG
`CCGAGGAAGA
`AGGAGATST:
`GTTCGA'.17GG
`
`.
`:::.
`K T
`5
`?
`7, 3 :
`' ~
`? F
`K
`L
`GGACTCCGAC
`C;CCCGTGCT
`AAGACCACGC
`GGACTCCGAC
`CTCCCGTGCT
`AAGACCACGC
`CC:'GAGGCTG
`JAGGGCACGA
`".';C:'GGTGCG
`CCTGAGGCTG
`GAGGGCACGA
`
`JGTGCG
`
`)
`:(
`G N
`W Q
`K
`.3:'~~ACAAGA
`GCAGGSG.7-A:
`.37G3ACAAGA
`:.n.C.::'G~TC~
`CGTCCCCTTG
`CGT:CACCGT
`CAC:7G7TC7
`
`E
`
`R
`
`'i
`5
`C
`K E
`S Y 7
`5
`•
`H E A
`14-1: ~:--::i-r:::,:..~ Ge:-:::;:-:;;._:- 4~ATGAGGC: :7.jCA:AAc: M::i.:...:;..c~CA. GAAGAG:::cr=
`5AAGAGCCTC
`A:TAZAC5CA
`144.
`5CATGAGG:7
`CTGCACAACC
`CAGAAGAG:A :GAG~:AC~A ::;TACTCCGA ~AC;:G7TG~ ~~A:'GTGC~T C~;CTCGGAG
`CTTCTCGGAG
`TGA7GTGCGT
`CAGAAGAGTA
`CGAG-.''AC7A
`GAr=7nTTG5
`CGTACTCCGA
`s
`~~:< .. A!...
`L,
`S i. S
`A
`CC3G~~.A.Z...:.: GGCCC~GA~: :;:~:TGGGGG
`:501 TCCCTG7C7C
`CC3G5AAAAT
`572CTGGGGG
`GGCCC7GA77
`AGSGACAGAG
`CCGGGAC7mA
`
`;_
`:.. L
`_-.. G
`•
`::;
`L L
`v A G
`CC7CCTGC::
`GCGT-:G::~G
`CCTCCTGC77
`GGAGGACGAA
` -CCA5C5GCC
`
`,,J
`
`5
`_ R V
`•
`l
`-
`I
`!
`K F S
`A F A
`F I
`~
`D
`A
`~
`F R V
`E
`R
`F
`~c:cccc:;:::;
`T7~A77~GG~ :AGGcn:C~7 C!TCAGAG:~ AAUi!CAGCA
`CGCCCCCGCG
`:561 TT:A7TGGGC
`GGAGCGCAGA
`7AGGCA7C77
`CTTCAGAG73
`AAGTTCAGCA
`~CGGGGGC~;
`.!\AG7AAC~CG A!CC~7A~AA GAAGTCTCAC :7:,;A~7~G:
`GCGGGGGC5:7
`AAG7AACCCG
`ATCCG7A5AA
`:.7.7C3C57C7
`GAAGTCTCAC
`77CAAG771G7
`
`E"IGURE : o
`FIGURE
`
`Miltenyi Ex. 1018 Page 4
`
`

`

`Patent Application Publication Jul. 1, 2004 Sheet 3 of 8
`l'atent Application Publication
`Jul. 1, 2004 Sheet 3 of 8
`
`US 2004/012636~1 A l
`US 2094/0126363 Al
`
`E s
`y N
`E L N
`'{ Q Q G
`L
`L G R R
`·Q N Q
`E L N
`L G
`L Y N
`R
`R
`G Q N Q
`E E
`'(
`Y
`Y Q Q
`1621 TACCAGCAGG GCCAGAACCA GCTCTATAAC GAGCTCAATC TAGGACGAAG AGAGGA.GTAC
`GAGCTCAATC
`GCTCTATAAC
`TAGGACGAAG AGAGGAGTAC
`1621 TACCAGCAGG
`GCCAGAACCA
`ATGGTCGTCC CGGTCTTGGT CGAGATATTG CTCGAGTTAG ATCCTGCTTC TCTCCTCATG
`CTCGAGTTAG
`CGAGATATTG
`ATCCTGCTTC TCTCCTCATG
`CGGTCTTGGT
`ATGGTCGTCC
`,,.. ..,
`p
`p E
`R R
`G R
`G
`:'I
`K
`i<
`D K R R
`D V
`L
`)
`P E M
`G G K ?
`R R K
`G R 3
`D V L
`K
`R
`R
`D
`1681 GATGTTTTGG ACAAGAGACG TGGCCGGGAC CCTGAGATGG GGGGAAAGCC GAGAAGGAAG
`GGGGAAAGCC
`CCTGAGATGG
`GAGAAGGAAG
`1681 GATGTTTTGG
`ACAAGAGACG
`TGGCCGGGAC
`CTACA.AAACC TGTTCTCTGC ACCGGCCCTG GGACTC'! ACC CCCCTTTCGG CTCTTCCTTC
`CCCCTTTCGG
`CTCTTCCTTC
`GGACTCTACC
`ACCGGCCCTG
`TGTTCTCTGC
`CTACAAAACC
`
`t
`A Y D
`-
`O K D K M A
`E G L Y N
`N ? Q
`E
`A Y D
`A
`M
`N E
`Q K D
`K
`I'
`N P
`E
`G
`L
`174 ~ AACCCTCAGG AAGGCCTGTA CAATGAACTG CAGAAAGATA AGATGGCGGA GGCCTACAGT
`AGATGGCGGA
`GGCCTACAGT
`CAGAAAGATA
`AAGGCCTGTA
`CAATGAACTG
`174k AACCCTCAGG
`TTGGGAG'J'C::t: T',-...CGGACA! GTTAC'tTGi-..C G'!'.:TTTCTA7 !CTACCGCCT CCGGATGTCA
`TCTACCGCCT
`CCGGATGTCA
`GTTACTTW,C
`GTZTTTCTAT
`T',,CGGACAT
`TTGGGAGTCC
`
`G K G H D G L Y Q G
`R R R
`I G M K' G £
`E
`D G L
`Y Q G
`H
`R R R
`G K G
`E I G
`M K' G E
`1801 GAGATTGGGA TGMAGGCGA GCGCCGGAGG GGCAAGGGGC ACGATGGCCT TTACCAGGGT
`ACGATGGCCT
`TTACCAGGGT
`GGCAAGGGGC
`1801 GAGATTGGGA
`GCGCCGGAGG
`TGAAAGGCGA
`CTCTAACCCT ACTTTCCGCT CGCGGCCTCC CCGTTCCCCG TGCTACCGGA AATGGTl.L:A
`CCGTTCCCCG
`TGCTACCGGA
`AATGGTcLCA
`CGCGGCCTCC
`ACTTTCCGCT
`CTCTAACCCT
`
`P P R
`L S T A T K D T Y D A L H M Q A L
`M Q A L
`P P R
`L S T
`T
`A
`T Y D
`A L H
`D
`K
`1861 CTCAGTACAG CCACCAAGGA CACCTACGAC GCCCTTCACA TGCAGGCCCT GCCCCCTCGC
`1861 CTCAGTACAG
`GCCCTTCACA
`TGCAGGCCCT
`GCCCCCTCGC
`CCACCAAGGA
`CACCTACGAC
`GAGTCATGTC GGTGGTTCCT GTGGATGCTG CGGGAAGTGT ACGTCCGGGA CGGGGGAGCG
`GAGTCATGTC
`GTGGATGCTG
`CGGGAAGTGT
`ACGTCCGGGA
`CGGGGGAGCG
`GGTGGTTCCT
`
`Not I
`Notl
`
`1921 TAAGCGGCCG C
`1921 TAAGCGGCCG C
`ATTCGCCGGC G
`ATTCGCCGGC G
`
`FIGURE lC
`FIGURE 1C
`
`Miltenyi Ex. 1018 Page 5
`
`

`

`Patent Application Publication
`Patent Application Publication Jul. 1, 2004 Sheet 4 of 8
`Jul. 1, 2004 Sheet 4 of 8
`bGh'1JAn·'t ;:_ .. :c~f1 ColEl
`Pad(7Sl )
`bdficlAn
`ColE1 Pad (751)
`.,
`
`US 2004/012636~1 Al
`US 2004/0126363 Al
`
`HyTk
`
`SpAn
`SpAn
`
`CMV Promoter
`C'M\I Promote r
`
`prAG-CD1 9R1HyTk
`
`lntron A
`Intro, A
`
`IRES
`
`CD19R scFvFc:Zeta
`
`Miltenyi Ex. 1018 Page 6
`
`

`

`Patent Application Publication
`Patent Application Publication Jul. I, 2004 Sheet 5 of 8
`Jul. 1, 2004 Sheet 5 of 8
`
`US 2004/012636:1 Al
`US 20144/0126363 Al
`
`hsp(FMC63)CD19R/pKEN
`hsp(FMC63)CD19R/pKEN
`09/27/00
`09/27/00
`anti-CD3zeta - overnight
`anti-CD3zeta - overnight
`· GAM-AP - 1 hour
`GAM-AP - 1 hour
`E;xposure - 1 hour
`Exposure -1 hour
`
`t(?
`,-
`'c5
`...
`
`i
`
`.,..
`....
`N
`'l5
`l
`
`. a ~
`
`~ ~
`C7I N ♦
`ID
`.- -
`0 UI
`,-
`:t: ., X
`t: 0 &!
`~ 0 <
`
`~ -
`
`---
`
`est eft 'Mr
`
`Miltenyi Ex. 1018 Page 7
`
`

`

`Patent Application Publication
`Patent Application Publication
`
`Jul. 1, 2004 Sheet 6 of 8
`Jul. 1, 2(N)4 Sheet 6 of 8
`
`US 2004/01 2636~1 A l
`US 2004/0126363 Al
`
`/11 vi"o Production of IFNy
`In vitro Production of 1FNly
`
`File· CD19>JD#8 of 12/6/99>CDl9(FMC63)pMG"·pac
`File CD19>Min of 12/6/99>CD19(FMC63)pMG'-pac
`IVS of2/8/00
`Elisa· 2/16/00
`IVS of 2/8/00
`Elisa: 2/16/00
`I O ~ I 06 each of responders or srimulato:s/well in 2 ml final. 24 well plate. 72 hr culture
`1.0 x 106 each of responders or stimulato..s/well in 2 ml final, 24 well plate. 72 hr culture
`f .,·m;:t
`F
`Rt!;ponders JD#B of 12/6/99 - CD 19 Clones: 86 (8T. W+. Fe-) and CI (8+.W+ .. Fe-).
`Responders' JD/$8 of 12/6/99 — CD I9 Clones: B6 (84-.W+, Fc-) and Cl (8+.W+.. Fc-).
`DHL-4, Parsons LCL,.
`Sthmulaton: Stimulator<. firTad to 8000 Rads) KS62,. ;)audi,. 1873-CRL\, JM-1 ,. DHL-4,. Parsons LCL,.
`Stimulators: Stimulators (irrad to 8000 Rads) K562,, Daudi,. 1873-CRL,,
`SVP-815,, lona/PMA Media
`SUP-B15,, lona/PMA. Media.
`
`Samples
`Samples
`
`I. B6/K562
`I. 86/K562.
`2. 86/Daudi.
`2. B6/Daudi,
`3. B6/1873-CRL,
`l . B6/187J-CRL.,
`4. B6/ JM-1,
`4. B6/ JM-1.
`.s. 8 6/ DHL-4.
`5. B6/DHL-4.
`6. B6/LCI.,
`6 B6/ LCL.
`7. 86/ SUP-81.S,
`7. 86/ SUP-B15,
`8. 86/1/P
`8. B6/UP
`9. 86/Mcdio.
`9. B6/Media
`10. CI/K562,
`_10. Cl/K562,
`II. CI /Daudi,
`II CI/Daudi,
`! 12. Cl/1873-CRL.
`12. Cl/1873-CRL,
`13. Cl/JM-I,
`' 13. Cl / JM-1.,
`1-1. Cl DHL--1
`14. CI DHL-4,
`15. Cu/ LCL.,,
`IS. Cl/ LCL,.
`16. Cl/ SUP-81.S,
`16. Cl/SUP-B15,
`17. Cl/ 1/P
`17. Cl/ VP
`18. Cl/Media
`18. Cl/ Media
`19. K562,
`19. 1<562,
`20. Daudi,
`-~-_!)audi,
`21. J 873-CRL.
`21. 1873-CRL,
`22. .1M-1,
`22. JM-1,
`23 DHL--1,
`23. DHL-4,
`24. LCL,
`24. LCL,
`25. SUP-B15,
`' 25. SUP-815,
`26. 1/P
`I 26. 1/P
`27. Media
`27. Media
`
`Diln
`Diln
`Plated
`Plated
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Near
`Neat
`Neat
`.at
`~-.:at
`1:10
`1: 10
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neal
`Neat
`Neat
`Neal
`Neat
`'.\.eJI
`Ne·
`Nc '
`Neat
`Neat
`Neat
`Neat
`Neat
`Neal
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neat
`Neal
`Neat
`Neat
`
`--'
`
`Units = extrapolated a ove
`b
`h. h
`d
`I!{ est standar
`Units' = extrapolated above highest standard
`n.,..1
`1 Diln
`TNFO Dilo
`Diln
`GM-CSF Diln
`EL-2
`Dila
`INFO
`GM-CSF
`Diln
`(0 2/m11 Plated
`(ne/ml)
`(ne/ml) Plated
`Plated
`(pg/mi)
`Plated
`(pg/ml)
`Plated
`(pg/m1)
`Plated
`Neat
`Neat
`0
`Neat
`0
`0
`Neat
`Neat
`0
`Neat
`0
`0
`1.10
`Neat
`0
`36
`Neat
`0
`Neat
`0
`1:10
`Neat
`0
`36
`1:10
`1:10
`0
`0
`0
`Neat
`Neal
`Neat
`113
`Neat
`113
`0
`1:10
`1:10
`0
`75
`0
`Neat
`Neat
`32
`Neat
`75
`Neat
`32
`1.10
`1.10
`0
`176
`Neat
`0
`0
`176
`Neat
`Neat
`Neat
`0
`1·10
`1-10
`0
`0
`Neat
`Ne-t
`71
`Neat
`Ne· t
`71
`85
`85
`1 10
`I 10
`Neat
`Neat
`U
`0
`Neat
`0
`Neat
`0
`116
`116
`1:10
`1:10
`37.155
`Neat
`803
`1·20
`37.155
`2231
` 803
`1.20
`2231
`Neat
`Neat
`Neat
`0
`Neat
`Neat
`0
`0
`Neat
`Neat
`0
`0
`0
`Neat
`Neat
`Neat
`34
`Neai
`34
`0
`0
`Neat
`Neat
`0
`0
`1:11)
`60
`60
`51
`1:10
`Neut
`51
`303
`Neat
`Neat
`303
`Neat
`}6
`!'JD
`60
`ND
`Neat
`36
`· 296
`Neat
`296
`Neat
`611
`~·e.11
`u
`1· 10
`,,
`1.10
`0
`Neat
`Ncat
`0
`Neal
`Neat
`25
`I)
`~ .. ~\
`25
`,.:~I
`I Ill
`Neat
`. (,9
`,1
`"
`1 III
`69
`\cat
`1:10
`4D
`1:10
`350
`40
`Neat
`Neat
`Neat
`0
`350
`0
`Near
`0
`1.10
`L:10
`0
`64
`Neat
`Neat
`Neat
`Neat
`6.S
`338
`338
`ND
`ND
`29.892
`1·20
`29.892
`1.10
`1:20
`5552
`301
`I 10
`301
`5552
`Neat
`0
`0
`Neat
`0
`0
`0
`Neat
`Neat
`Neat
`0
`Neal
`0
`Neat
`Neat
`Neat
`0
`Neat
`0
`0
`0
`0
`0
`Neal
`Neat
`0
`0
`Neat
`Neat
`0
`0
`0
`n
`n
`Neal
`73
`Neal
`0
`Neat
`Neat
`73
`0
`n
`NeQI
`0
`Neat
`Neat
`0
`Neat
`0
`0
`0
`Io
`Neat
`II
`Neat
`Neat
`0
`Neat
`II
`0
`(I
`u
`0
`Neat
`0
`Neat
`Neat
`0
`0
`ll
`Neal
`.
`u
`,J
`Neat
`0
`13
`0
`0
`, Neat
`Neat
`1 Neat
`0
`0
`Neat
`0
`Neat
`Neat
`Nellt
`0
`0
`0
`
`_
`
`IFNy
`IFNy
`fno/ml\ 1
`(pg/m1)
`25
`25
`1
`l
`1362
`1362
`·1
`967
`967
`396
`396
`645
`645
`1245
`1!45
`946
`946
`>10.001)
`>10.000
`0
`0
`0
`0
`1679
`1679
`
`J
`•
`
`631
`631
`li~II
`69)1
`'483
`•~SJ
`1245
`1245
`
`0
`
`-
`
`~
`
`'
`!
`l
`I
`J
`1
`I
`I
`I
`I
`
`i
`!
`
`F trArt, 4
`
`Miltenyi Ex. 1018 Page 8
`
`

`

`0
`N
`
`r -u E ·
`
`Patent Application Publication
`
`}it Jo L WHIS
`
`US 2004/0126363 Al
`
`'Is
`
`~ z
`
`ft_
`
`0
`0
`co
`
`IV)
`
`.;)
`~
`
`.._
`0
`C>I
`0
`~ C.,
`.._,,,,
`ffl
`E
`E
`co
`0,
`z ~
`I
`0
`,~l
`0
`(0 LL
`ll
`
`IFN--y
`
`I.O
`
`CD
`I a.
`::,
`(/)
`
`~
`
`_J
`0
`...J
`
`'V ~
`I
`....J ~
`-,
`I
`0
`
`I
`
`_J
`~
`u
`t--co
`
`I
`('I")
`
`~
`
`SUP-B15
`
`0
`'V
`
`0
`0
`N
`
`0
`
`-0 N
`(0
`:,
`co ~
`LO
`0
`
`co ·-'-0
`Q)
`~
`
`0 -
`
`LCL
`
`DHL-4
`
`JM-1
`
`1873-CRL
`
`Daudi
`
`K562
`
`Media
`
`0
`
`200
`
`400
`
`Fr lace, S-
`
`-I
`_
`
` 1
`1400 1600
`100C 120(
`800
`600
`IFN-gamma (pg/ml)
`
`Miltenyi Ex. 1018 Page 9
`
`

`

`Patent Application Publication
`Patent Application Publication
`
`Jul. 1, 2004 Sheet 8 of 8
`Jul. 1, 2004 Sheet 8 of 8
`
`US 2004/01263621 Al
`US
`2004/0126363 Al
`
`Tuaet
`Target
`187lCIU.
`1873 CRI.
`JM·I
`1M-1
`S..oBl.5
`Sup1315
`K.562
`K362
`
`77.1 ' 69.5
`
`Eflco,o,: CDl9 IFMC631 Clon
`Effector C019 IFSIC63) Clon
`5: 1
`25: 1
`50:1
`25:1
`SO:1
`·s6: 2
`77.1 I 68.5 • 66.2
`87.6
`83.9
`87.6
`87.6
`87.6
`83.9
`87.7
`85.4
`94.8
`94.8
`87.7
`85.4
`4 ,6
`7 .7
`6 ,6
`4.6
`7.7
`6.6
`
`-
`I -
`1
`1: 1
`1. 1
`8P.!a
`·ss.~
`55.2
`se.s
`56.8
`1.6
`1.6
`
`,,,
`
`(j1fector: C019 (FMC631 CJona C12
`Wm*. 0019 (FMCS!' Clone C12
`5: 1
`25:1
`SO: 1
`50.1
`25:1
`5:1
`1:1
`64. 1
`4!>.8
`69. 3
`86.3
`45.8
`89.3
`88.3
`64.1
`77.4
`S7.4
`80.S
`84. 9
`57.4
`80.5
`84.9
`77.4
`85.9
`58.2
`89.9
`83. 5
`89.9
`83.5
`95.9
`58.2
`5.2
`4.9
`3.0
`1.2
`5.2
`4.9
`3.0
`1.2
`
`T.,get
`Tw CI
`873 CRL
`1873 CRL
`JM-1
`AAA
`S..pBU
`Su B15
`U6l
`K362
`
`Tuoet
`Tappet
`1873 CIU.
`1873 CM.
`JM-1
`3N4-1
`SuoBIS
`SupB15
`IU6l
`K562
`
`£tifec1or: C019 IFMC83J Clon• E8
`ftleetw: CD19 IFMC63) Clam 88
`S: I
`1:1
`25: 1
`~iO:l
`25:1
`5:1
`1:1
`50:1
`79.5
`81.6
`80.7
`8 1.0
`81.0
`79.5
`61.6
`80.7
`110.0
`611.4
`90. 5
`84, I
`80.0
`66.4
`84.1
`90.5
`74.3
`93.&
`9 3.3
`97.8
`93.6
`74.3
`97.6
`93.3
`0 .0
`0.4
`0 .0
`0 .0
`0.4
`0.0
`0.0
`0.0
`
`-
`
`-
`-
`-
`011•14.c.. Cm. C..
`
`Itt
`to
`50
`0
`40
`
`47
`
`:1
`
`'1
`
`10t
`
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`Miltenyi Ex. 1018 Page 10
`
`

`

`US 2004/0126363 Al
`US 2004/0126363 Al
`
`J ul. 1, 2004
`Jul. 1, 2004
`
`1
`
`CD19-SPE◄ClFlC RIWrRECT ED IMMUNE CELLS
`CD19-SPECIFIC REDIRECTED IMMUNE CELLS
`
`CROSS-REFERENCE 1'0 RELATED
`CROSS-REFERENCE TO RELATED
`APPLICATION
`APPLICATION
`
`[<H)0l] This applicalioa claims priority 10 Provisional
`[0001] This application claims priority to Provisional
`t\pplil:a1ioa Seriru No. 60/246, 11 7, fileJ Nov. 7, 2000, the
`Application Serial No. 60/246,117, filed Nov. 7, 2000, the
`disclosure of which is incorporated by reforeace.
`disclosure of which is incorporated by reference.
`
`BACKGROUND OF THE [NVENTION
`BACKGROUND OF THE INVENTION
`[0002] Tb.is invention rclalcs to the field of gcnclically
`[0002] This invention relates to the field of genetically
`engim:ercd, redircc!t:!d immune cells and to the field of
`engineered, redirected immune cells and to the field of
`cellular immunorbcrapy of 8 -cell malignancies. B-cell lym(cid:173)
`cellular immunotherapy of B-cell malignancies, B-cell lym-
`phoproliferative syadromes and B-cell mediaced au1oim(cid:173)
`phoproliferative syndromes and B-cell mediated autoim-
`mune diseases. The publications and other materials used
`mune disei1$c,s. The publications and other materials used
`herein to il111min3k the background of the invenl ion or
`herein to illuminate the background of the invention or
`provide additional details respecting the practice arc incor(cid:173)
`provide additional details respecting the practice are incor-
`porated by reference.
`porated by reference.
`[0003] Approximately half of all bcmatopoictic stem t:cll
`[0003] Approximately half of all hernatopoietic stem cell
`traosplanlatioo (HSC') procedures performed io tbt' Uoited
`transplantation (HSC) procedures performed in the United
`States are for the trna1men1 of hematologic rnn.lignaocy [1 ].
`States are for the treatment of hematologic malignancy [1].
`Tbc initial abstadl,'S for succ~ssflll HSC transplantation
`The initial obstacles for successful HSC transplantation
`were ia lnrge part due to inadequate lrealmenl modalities for
`were in large part due to inadequate treatment modalities for
`amclirm1ti11g regimen-related toxicities and for cootrollin_g
`ameliorating regimen-related toxicities and for controlling
`infections and gra(t-versus-lmJ;I disease
`opportUJJistic
`opportunistic
`infections and graft-versus-host disease
`(GVHD) [2-5]. /\s supportive care measures have improved
`(GVHD) [2-5]. As supportive care measures have improved
`over 1bc last dccude, post-lraosplant disease relapse bas
`over the last decade, post-transplant disease relapse has
`emerged as tbtc major impediment to improving 1bc outcome
`emerged as the major impediment to improving the outcome
`of Ibis pa1icnt. popu la1ion [G-10), The inability of rnaxi.mally
`of this patient population [6-10]. The inability of maximally
`iateosive prepar;11ive regimens combintd with immunologic:
`intensive preparative regimens combined with immunologic
`graft-versus-tumor reactivity to eradicate minimal residual
`graft-vtrsus-tumor reactivity to eradieale minim.ti residual
`disease is th.: mechanism of lrca1men1 failure in allogeneic
`disease is the mechanism of treatment failure in allogeneic
`transpluntalion while. in lhc aulologous selling, tumor con(cid:173)
`transplantation while, in the autologous setting, tumor con-
`tamination o.f lbe stem cell graft can ~lso contribute lo
`tamination of the stem cell graft can also contribute to
`posHransplant relapse [ 11). Targeting minimal residual dis(cid:173)
`post-transplant relapse [II]. Targeting minimal residual dis-
`ease early af11~r transplantation is one stralcgy to conso!ichile
`ease early after transplantation is one strategy to consolidate
`tile lumor cytorodut:tion acbievod witb rnyeloahl,ttive pro(cid:173)
`the tumor cytoreduction achieved with mycloablative pre-
`paralive regimens and pUige, in vivo, malignant cells trans(cid:173)
`parative regimens and purge, in vivo, malignant cells trans-
`ferred witb aulologous stem cell grafts. The utility of thera(cid:173)
`ferred with autologous stem cell grafts. The utility of thera-
`peutic modalitit::s for targeting minimal residual disease
`peutic modalities for targeting minimal residual disease
`shortly following stem cell rescue is depeadent on both a
`shortly following stem cell rescue is dependent on both a
`limited i;pcctru01 of toxicity and the susceptibilit.y of residual
`limited spectrum of toxicity and the susceptibility of residual
`tumor cells 110 the modality's antilumor effector mccha(cid:173)
`tumor cells to the modality's antitumor effector mecha-
`uism(s). The successful elimination of pcrsis1en1 minimal
`nism(s). The successful elimination of persistent minimal
`residual dise::a.sc .should not unly have a major impac1 on tbe
`residual disease should not only have a major impact on the
`uu1come of trnnspluntation for hematologic malignancy uti(cid:173)
`outcome of transplantation for hematologic malignancy uti-
`lizing cu1rca1 myeloabla1ive preparative regimens but may
`lizing current mycloablative preparative regimens but may
`also providt opportunities to decreaSt: the iotcnsi1y of 1hes~
`also provide opportunities to decrease the intensity of these
`regimens aod their attendant loxicities.
`regimens and their attendant toxicities.
`[0004] TI1e prognosis for patients wi1h bcr-abl positive
`[0004] The prognosis for patients with bcr-ahl positive
`Acute Lymphioblaslic Leukemia (ALL) trcaled with chemo(cid:173)
`Acute Lymphoblastic Leukemia (ALL) treated with chemo-
`therapy is poor and allogeneic transpla111ation has offered a
`therapy is poor and allogeneic transplantation has offered a
`curative option for many patients when an appropriate donor
`curative O[Jlio10 for m11oy paticots when an approprialc clooor
`was available. For example at the City of (lope. 76 patients
`was ttvailable. For ex!I.Olple al the City of !lope, 76 putients
`with bcr-abl positive ALL were treated with allogeneic Bone
`with bcr-abl positive ALL W!lrtl treated with :iUogcneic 130111!
`M,1rrow Tra.n:;planla.lion ( 13MT) Crom a I I LA matched donor.
`Marrow Transplantation (BMT) from a LILA matched donor.
`Of these palients, 26 were in first remission, 35 were
`Of these patients, 26 were in first remission, 35 were
`transplanted after first remission. The two year probability of
`traasvluoted ~1ftcr first remission. The two year probahiUty of
`disease free survival was 68% with a 10%