W0 98/4533]
`
`I'C‘I‘IUSS’li/Uooml
`
`-
`
`lower VEGF concentration. The result therefore supports the conclusion that the affinity-
`
`improved variant is at least 30-fold improved in affinity for VEGF, and that it effectively
`
`blocks VEGF activity in vitro. Since the variant YO3 l7 differs from YO313-l only in the
`
`reversion of the VLl sequence to wild-type (Fig. 10A), it is predicted that Y0317 will have
`
`similar activity to YO3 13-1.
`
`Variant Y03 l 7 (Fab) and humanized variant F(ab)-12 fi‘om Example 1 (full length and
`
`Fab) were compared for their ability to inhibit bovine capillary endothelial cell proliferation
`
`in response to a near maximally effective concentration of VEGF using the assay described
`
`’ in Example 1. As illustrated in Figure 12, YO317 was markedly more effective at inhibiting
`
`10
`
`bovine capillary endothelial cell proliferation than the full length and Fab forms of F(ab)-12
`
`in this assay. The YO3l7 affinity matured Fab demonstrated an EDSO value in this assay
`
`which was at least about 20 fold lower than F(ab)-12 Fab.
`
`
`
`..3-L‘J.-.;_-.--___
`
`
`
`
`
`
`
`Regeneron Exhibit 1024.0801
`
`78
`
`

`

`WO 98/45331
`
`PCT/US98/06604
`
`WHAT I
`
`LAIMED IS:
`
`1.
`
`A humanized anti-VEGF antibody which binds human VEGF with a Kd value of no
`
`more than about 1 x 10'3M.
`
`2.
`
`A humanized anti-VEGF antibody which binds human VEGF with a Kd value of no
`
`more than about 5 x 10'9M.
`
`3.
`
`' A humanized anti-VEGF antibody which has an EDSO value of no more than about
`
`SnM for inhibiting VEGF-induced proliferation of endothelial cells in vitro.
`
`10
`
`15
`
`20
`
`4.
`
`A humanized anti-VEGF antibody which inhibits VEGF—induced angiogenesis in vivo.
`
`5.
`
`The humanized anti-VEGF antibody of claim 4 wherein 5mg/kg of the antibody
`
`inhibits at least about 50% of tumor growth in an A673 in vivo tumor model.
`
`6.
`
`The humanized anti-VEGF antibody of claim I having a heavy chain variable domain
`
`comprising the following hypervariable
`
`region amino acid
`
`sequences: CDRHI
`
`(GYX,FTX2YGMN, wherein X1 is T or D and X2 is‘N or H; SEQ ID N01128), CDRH2
`
`(WINTYTGEPTYAADFKR; SEQ ID N022) and CDRH3 (YPX1YYGX28HWYFDV,
`
`wherein X] is Y or H and Xzis S or T; SEQ ID NO: 129).
`
`7.
`
`The humanized anti-VEGF antibody of claim 6 comprising the amino acid sequence
`
`of SEQ ID N027.
`
`8.
`
`The humanized anti-VEGF antibody of claim 6Vhaving a heavy chain variable domain
`
`25
`
`COmprising the following hypervariable
`
`region amino acid sequences: CDRHl
`
`(GYTF'I'NYGMN; SEQ ID N021), CDRHZ (WINTYTGEPTYAADFKR; SEQ ID N02)
`
`and CDRH3 (YPHYYGSSHWYFDV; SEQ ID N03).
`
`79
`
`Regeneron Exhibit 1024.0802
`
`
`
`

`

`W0 98/4533!
`
`-
`
`PCT/US98/Ubou4
`
`9.
`
`The humanized anti-VEGF antibody of claim I having a light chain variable domain
`
`comprising the
`
`following hypervariable
`
`region
`
`amino acid sequences: CDRLI
`
`(SASQDLSNYLN; SEQ ID N014), CDRLZ (FTSSLHS; SEQ ID N025) and CDRL3
`
`(QQYSTVPWT; SEQ ID NO:6).
`
`10.
`
`The humanized anti-VEGF antibody of claim 9 comprising the amino acid sequence
`
`ofSEQ ID N018.
`
`11.
`
`' The humanized anti-VEGF antibody of claim 1 having a heavy chain variable domain
`
`10
`
`comprising the amino acid sequence of SEQ ID N027 and a light chain variable domain
`
`comprising the amino acid sequence of SEQ ID N018.
`
`12.
`An anti-VEGF antibody light chain variable domain comprising the amino acid
`sequence:
`I
`
`15 DIQX,TQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLH
`SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKR (SEQ
`
`ID N01124), wherein X| is M or L.
`
`13.
`
`An anti-VEGF antibody heavy chain variable domain comprising the amino acid
`
`20
`
`sequence:
`
`'
`
`EVQLVESGGGLVQPGGSLRLSCAASGYX,FTXzYGMNWVRQAPGKGLEWVGWI
`
`NTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPX3YY
`
`GX4SHWYFDVWGQGTLVTVSS (SEQ ID NO:125), wherein X1 is T or D; Xzi is N or H;
`
`X3 isYorHandX4 isSorT.
`
`25
`
`14.
`
`A variant ofa parent anti-VEGF antibody, wherein said variant binds human VEGF
`
`- -.and ,comprisesan amino acid substitution in a hypervan'able region of a heavy chain variable
`
`domain of said parent antibody.
`
`30
`
`15.
`
`The variant of claim I4 wherein said parent antibody is a human or humanized
`
`antibody.
`
`80
`
`Regeneron Exhibit 1024.0803
`
`
`
`
`
`.._T______n_'_;__“HA___"“3.
`
`

`

`W0 98/4533]
`
`‘
`
`PCI'IUS98/uooo4
`
`16.
`
`The variant of claim 14 which binds human VEGF with a Kd value of no more than
`
`about 1 x 10'8M.
`
`t
`
`17.
`
`The variant of claim 14 which binds human VEGF with a Kd value of no more than
`
`about 5 x 10'9M.
`
`18.
`
`The variant of claim 14 wherein the substitution is in CDRHl of the heavy chain
`
`variable domain.
`
`10
`
`19.
`
`The variant of claim 14 wherein the substitution is in CDRH3 ofthe heavy chain
`
`variable domain.
`
`20.
`
`The variant of claim l4 which has amino acid substitutions in both CDRHI and
`
`CDRH3.
`
`15
`
`‘ 21.
`
`The variant of claim 14 which binds human VEGF with a Kd value less than that of
`
`said parent antibody.
`
`22.
`
`The variant of claim 14 which has an EDSO value for inhibiting VEGF-induced
`
`20
`
`proliferation of endothelial cells in vitro which is at least about 10 fold lower than that of
`
`said parent antibody.
`
`23.
`
`The variant of claim l8 wherein the CDRHl comprises the amino acid Sequence:
`
`GYDFTHYGMN (SEQ ID NO: 126)
`
`‘25
`
`24.
`
`The variant of claim 19 wherein the CDRH3 comprises the amino acid sequence:
`
`YPYYYGTSHWYFDV (SEQ ID NO: 127).
`
`25.
`
`The variant of claim 14 wherein the heavy chain variable domain comprises the amino
`
`30
`
`acid sequence of SEQ ID NO: I I6.
`
`Regeneron Exhibit 1024.0804
`
`

`

`WU 95/4DJJI
`
`l‘L' I [USHKIUOOU‘I
`
`5
`
`:1]
`
`26.
`
`The variant of claim 25 further comprising the light chain variable domain amino acid
`
`sequence of SEQ ID NO: 124.
`
`o
`
`27.
`
`The variant of claim 26 cOmprising the light chain variable domain amino acid
`
`5
`
`sequence of SEQ ID NO}! 15.
`
`28.
`
`The humanized anti-VEGF antibody of claim I which is a full length antibody.
`
`29.
`
`The humanized anti-VEGF antibody of claim 28 which is a human IgG.
`
`10
`
`30.
`
`The humanized anti-VEGF antibody of claim I which is an antibody fragment.
`
`3].
`
`The antibody fragment of claim 30 which is a Fab.
`
`15
`
`32.
`
`A composition comprising the humanized anti—VEGF antibody of claim 1 and a
`
`pharmaceutically acceptable carrier.
`
`33.
`
`A composition comprising the variant anti-VEGF antibody of claim 14 and a
`
`pharmaceutically acceptable carrier.
`
`20
`
`l
`
`34.
`
`Isolated nucleic acid encoding the antibody of claim 1.
`
`35.
`
`A vector comprising the nucleic acid of claim 34.
`
`m 25
`
`36.
`
`A host cell comprising the vector of claim 35.
`
`i <7
`
`2
`
`- . , i “37.“ 7 A process ofproducing a humanized anti-VEGF antibody comprising culturing the
`host cell ofclaim 3650 that the nucleic acidisflekpres'séd’f’ T '
`“ " ~ -
`7-
`
`..
`
`- a
`
`30
`
`38.
`
`The process of claim 37 further comprising recovering the humanized anti-VEGF
`
`antibody from the host cell culture.
`
`
`
`Regeneron Exhibit 1024.0805
`
`

`

`WO 98/45331
`
`'
`
`PCTIUS98/06604
`
`39.
`
`A method for inhibiting VEGF-induced angiogenesis in a mammal comprising
`
`administering a therapeutically effective amount of the humanized anti-VEGF antibody of
`
`claim 1 to the mammal,
`
`40.
`
`The method of claim 39 wherein the mammal is a human.
`
`41.
`
`The method of claim 39 wherein the mammal has a tumor.
`
`42.
`
`’ The method of claim 39 wherein the mammal has a retinal disorder.
`
`Regeneron Exhibit 1024.0806
`
`

`

`W0 98/453251
`
`PCI'IUS98/Uoou4
`
`1/16
`
`1
`
`°
`
`1 O
`
`2 0
`
`3 0
`
`4 O
`
`A4.6.1
`EIQLVQSGPELKQPGETVRISCKASGYTFTNYGMNWVKQA
`-k
`*
`* *
`it
`* * *
`*
`*
`”I:
`F(ab)-12 EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQA
`* ****
`
`humIII
`
`EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQA
`
`A46. 1
`
`PGKGLKWMGWINTYTGEPTYAADFKRRFTFSLETSASTAYL
`*
`*
`*
`*
`
`5 O
`
`a
`
`6 O
`
`7 O
`
`8 0
`
`F(ab)-12 PGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYL
`** **** ***
`*** *
`* * *
`*
`*
`
`humIII
`
`PGKGLEWVSVI SGDGGSTTYADSVKGRFTI SRDNSKNTLYL
`
`A4.6.l
`
`abc
`90
`110
`QISNLKNDDTATYFCAKYPHYYGSSHWYFDVWGAGTTVTVSS
`* ~k *
`~k * *
`*
`*
`~k
`*
`
`(SEQ.ID N029)
`
`F(ab)-12 QMNSLRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSS
`*
`*
`
`(SEQ.ID NO:7)
`
`humIII QMNSLRAEDTAVYYCARG— - ~ — — — - - - - FDYWGQGTLVTVSS
`
`(SEQ.ID N021 1)
`
`'
`
`FIG._1A
`
`a
`
`I i E
`
`i Il iI
`
`‘
`
`;§"
`
`3-"
`
`i
`
`
`
`
`
`..._._.*__....__~_..._...____
`
`_
`'
`
`
`
`‘...‘E.‘..._~.-_.......P‘..
`
`3
`!
`
`l
`
`1 O
`
`2 0
`
`3 0
`
`4 O
`
`A4.6.1
`
`DIQMTQTTSSLSASLGDRVIISCSASQDISNYLNWYQQKP
`* *
`*
`*
`*
`
`F(ab)- ] 2 DIQMTQS PS SLSASVGDRV‘I‘I‘I‘C SASQDI SNYLNWYQQKP
`' *
`*
`*
`
`humKI DIQMTQSPS SLSASVGDRVTITCRASQS I SNYLAWYQQKP
`
`a
`
`A4.6.1
`
`8 o
`-
`7 o
`' 60
`5 o
`DGTVKVLIYFTSSLHSGVPSRFSGSGSGTDYSLTISNLEP
`* *
`* * * *
`* *
`
`F(ab)— 12 GKAPKVL IYFTSSLHSGVPSRFSGSGSGTDFTLTI SSLQP
`*
`*-k
`*
`
`humKI GKAPKLLIYAAS’SLESGVPSRFSGSGSGTDF-TLTISSLQP ,,
`
`A46. 1
`
`1 O 0
`9 O
`EDIATYYCQQYSTVPWTFGGGTKLEIKR
`*
`‘k
`*
`
`(SEQID NO: 10)
`
`F(ab)-12 EDFATYYCQQYS’I‘VPWTFGQGTKVEIKR
`***
`
`(SEQJD NO:8)
`
`humKI
`
`EDFATYYCQQYNSLPWTFGQGTKVEIKR
`
`(SEQID N0212)
`
`FIG._ 1 B
`
`SUBSTITUTE SHEET (RULE 25)
`
`Regeneron Exhibit 1024.0807
`
`

`

`WO 98/45331
`
`PCT/US98/06604
`
`2/16
`
`N
`
`
`6E
`
`SUBSTITUTE SHEET (RULE 25)
`
`Regeneron Exhi
`
`it 1024.0808
`
`

`

`WU 98/4555l
`
`'
`
`rt. l IUD'VGIU 004
`
`180000
`
`3/16
`
`I NO ADDITION
`
`
`E ,
`
`*—
`
`I 9 u
`
`! A
`3 E
`0: 3
`N (3
`':§ -
`D
`'—
`
`CONTROL
`MAb
`FIG. -4 (5)
`
`muMAb
`VEGF
`(0.5)
`
`rhuMAb
`VEGF
`(0.5)
`
`rhuMAb
`VEGF
`(5)
`
`SUBSTITUTE SHEET (RULE 26)
`
`Regeneron Exhibit 1024.0809
`
`40— VEGF + muMAb VEGF
`
`_D_ VEGF + rhuMAb VEGF
`. VEGF
`
`
`
`j
`2' m .
`:35:
`LU
`3:53
`*- cL
`(>0),
`0 .1
`Z _l
`uJuJ“
`0
`
`.140000
`
`100000
`
`60000
`1
`
`20000
`
`FIG. _ 3 0
`-
`
`on
`
`1
`
`10
`
`100
`
`1000
`
`10000
`
`MAbCONCENTRAHONugmm
`
`

`

`WU 93/433.“
`
`fl. I lUD’E/UOOUQ
`
`4/16
`
`A4.6.|
`
`hu2.0
`
`hu2. IO
`
`A4.6.l
`
`hu2.0
`
`40
`_
`30
`20
`‘_
`10
`.
`DIQMTQTTSSLSASLGDRVIISCSASQDISNYLNWYQQKP
`'k
`'
`**
`*
`*
`
`DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKP
`
`DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKP
`
`8O
`70
`60
`50
`DGTVKVLIYFTSSLHSGVPSRFSGSGSGTDYSLTISNLEP
`**** *
`*‘k
`*
`*
`
`GKAPKLLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQP
`
`hu2. IO
`
`GKAPKLLIYFTSSLHSGVPSRFSGSGSGTDYTLTISSLQP
`
`A4.6.]
`
`hu2.0
`
`hu2.10
`
`90
`
`100
`
`'
`
`EDIATYYCQQYSTVPWTFGGGTKLEIK
`*
`*
`*
`
`(SEQJDPKIIO)
`
`EDFATYYCQQYSTVPWTFGQGTKVEIK
`
`(SEQJDI«3J3)
`
`EDFATYYCQQYSTVPWTFGQGTKVEIK
`
`(SEQJDPflIlS)
`
`FIG._5A
`
`A4.6.l
`
`hu2.0
`
`hu2. 10
`
`10
`
`~
`
`20
`
`3O
`
`40
`
`EIQLVQSGPELKQPGETVRISCKASGYTFTNYGMNWVKQA
`*
`‘k
`** *
`*** *
`*
`*
`
`EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQA
`
`EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWIRQA
`
`A4.6.1
`
`80
`70
`60
`50 a
`PGKGLKWMGWINTYTGEPTYAADFKRRFTFSLETSASTAYL
`*
`.k
`* *** *4:
`-k
`
`hu2.0
`
`PGKGLEWVGWINTYTGEPTYAADFKRRFTISRDNSKNTLYL
`
`hu2.10
`
`PGKGLEWVGWINTYTGEPTYAADFKRRFTISLDTSASTVYL
`
`A4.6. I
`
`hu2.0
`
`hu2.10
`
`110
`‘
`lOOabcdef
`90
`abc
`QISNLKNDDTATYFCAKYPHYYGSSHWYFDVWGAGTTVTVSS
`*~k* ***
`*
`‘k
`‘k
`'k
`*
`
`. (SEQID NO:9)
`
`QMNSLRAEDTAVYYCARYPHYYGSSHWYFDVWGQGTLVTVSS
`
`(SEQID NO:14)
`
`QMNSLRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSS
`
`(SEQID N0216)
`
`‘ FIG._EB
`
`SUBSTITUTE cuczr (m u I: on
`
`Regeneron Exhibit 1024.0810
`
`
`
`

`

`W0 98/4533]
`
`I‘C'l'l US98/UOOU4
`
`5/16
`
`
`
`
`
`FIG.:_6
`
`SUBSTITUTE SHEET (RULE 26)
`
`Regeneron Exhibit 1024.0811
`
`

`

`a
`
`W0 98/4533]
`
`PCT[U898/06604
`
`
`
`Ampr
`
`pBR322 ori
`
`TRANSFORM E. coli
`
`
`
`+M13KO7 HELPER PHAGE
`
`FAB-p111 FUSION
`/
`
`
`
`PHAGEMI
`
`
`
`FIG._ 7
`
`an l—nc-I—n .—_ -. ._-_ .-—.--
`
`Regeneron Exhibit 1024.0812
`
`

`

`
`
`
`
`#4.;z—‘v____..._...__._._;.._..,‘..-4.‘;_._l‘...____......_.___
`
`
`
`
`
`W0 98/4533]
`
`PCT/US98/06604
`
`GAATTCAACT TCTCCATACT
`
`7/16
`AAATACAGAC
`TTGGATAAGG
`
`ATGAAAAATC TCATTGCTGA
`
`60
`
`GTTGTTATTT.AAGCTTTGGA
`
`GATTATCGTC
`
`ACTGCAATGC
`
`TTCGCAATAT GGCGCAAAAT
`
`GACCAACAGC GGTTGATTGA
`
`TCAGGTAGAG
`
`GGGGCGCTGT
`
`ACGAGGTAAA GCCCGATGCC
`
`120
`
`180
`
`AGCATTCCTG ACGACGATAC
`
`GGAGCTGCTG
`
`CGCGATTACG
`
`TAAAGAAGTT ATTGAAGCAT
`
`240
`
`CCTCGTCAGT AAAAAGTTAA
`
`TCTTTTCAAC
`
`AGCTGTCATA
`
`AAGTTGTCAC GGCCGAGACT
`
`300
`
`TATAGTCGCT TTGTTTTTAT
`
`TTTTTAATGT
`
`ATTTGTAACT
`
`AGAATTCGAG CTCGGTACCC
`
`360
`
`'GGGGATCCTC TAGAGGTTGA
`
`GGTGATTTT ATG AAA AAG
`
`Met Lys Lys
`—23
`
`ATT GCT ACA
`Ile Ala Thr
`—5
`
`AAT ATC GCA TTT CTT
`Asn Ile Ala Phe Leu
`—20
`
`413
`
`AAC GCG TAC GCT GAT
`
`461
`
`Asn Ala Tyr Ala Asp
`1
`
`CTT GCA TCT ATG TTC GTT TTT TCT
`Leu Ala Ser Met Phe Val Phe Ser
`—15
`—10
`BEGIN stII
`TCC CCG AGC
`Ser Pro Ser
`
`CAG TTG ACC
`Gln Leu Thr
`5
`
`CAG
`Gln
`
`ATC
`Ile
`
`ACC
`Thr
`
`AGG
`
`Arg
`
`GTC ACC ATC
`Val Thr Ile
`20
`BEGIN LIGHT CHAIN
`AAC TGG TAT CAA CAG
`
`TGC AGC GCA AGT CAG GAT
`
`Cys Ser Ala
`25
`
`Ser Gln Asp
`
`ATT AGC AAC TAT TTA I
`Ile Ser Asn Tyr Leu
`30
`
`AAA CCA GGA
`
`AAA GCT CCG
`
`AAA CTA CTG ATT TAC
`
`SIGNAL SEQUENCE
`TCC CTG TCC
`GCC TCT GTG GGC GAT
`Ser Leu Ser
`10
`
`Ala Ser Val Gly Asp
`15
`
`509
`
`557
`
`605
`
`Asn Trp Tyr Gln Gln
`35
`
`Lys Pro Gly
`40
`
`Lys Ala Pro
`
`Lys Leu Leu Ile Tyr
`45
`
`ACC TCC TCT
`Thr Ser Ser
`
`CTC
`Leu
`
`TTC
`Phe
`50
`
`CAC TCT GGA
`
`His Ser Gly
`55
`
`GTC CCT TCT
`Val Pro Ser
`60
`
`CGC TTC TCT GGA TCC
`
`653
`
`Arg Phe Ser Gly Ser
`65
`
`GGT
`
`TCT GGG ACG
`
`GAT
`
`TAC ACT CTG
`
`Gly
`
`Ser Gly Thr
`
`Tyr Thr Leu
`
`Asp
`70
`
`ACC ATC AGC
`
`AGT CTG CAG CCA GAA
`Thr Ile Ser Ser Leu Gln Pro Glu
`75
`80
`
`701
`
`GAC
`
`Asp
`
`TTC GCA ACT
`Phe Ala Thr
`85
`
`TAT
`
`TAC TGT CAA
`
`CAG TAT AGC
`
`ACC GTG CCG TGG ACG
`
`749
`
`Tyr
`
`Tyr Cys Gln
`
`Gln Tyr Ser Thr Val Pro Trp Thr
`90
`95
`
`TTT
`Phe
`
`TCT
`Ser
`
`GCT
`Ala
`130
`
`GGA CAG GGT
`
`Gly Gln Gly
`’“ 100* w
`
`ACC
`Thr
`
`AAG GTG GAG
`
`ATC AAA CGA
`
`ACT GTG GCT GCA CCA
`
`Lys Val Glu
`“IA,,,_,105_
`
`Ile Lys Arg Thr Val Ala Ala Pro
`51150,
`,
`
`797-
`
`GTC TTC ATC
`Val Phe Ile
`115
`
`TTC
`Phe
`
`CCG CCA TCT
`Pro Pro Ser
`120
`
`GAT GAG CAG
`
`TTG AAA TCT GGA ACT
`
`845
`
`Asp Glu Gln Leu Lys Ser Gly Thr
`125
`
`TCT GTT GTG
`Ser Val Val
`
`TGC
`
`Cys
`
`CTG CTG AAT
`Leu Leu Asn
`135
`
`AAC TTC TAT CCC AGA GAG GCC AAA
`
`893
`
`Asn Phe Tyr Pro Arg Glu Ala Lys
`140
`145
`
`Hall/4X
`
`-.-—__._- .__ ‘. ._.__ 1-... — -A‘
`
`Regeneron Exhibit 1024.0813
`
`

`

`W0 98/45331
`
`PCT/US98/06604
`
`7
`8/ 16
`GTA CAG TGG AAG GTG GAT AAC GCC CTC CAA TCG GGT AAC TCC CAG ~GAG
`Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
`150
`155
`160
`
`AGT GTC ACA GAG CAG GAC AGC AAG GAC AGC ACC TAC AGC CTC AGC AGC
`Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
`165
`.
`170
`175
`
`941
`
`989
`
`ACC CTG ACG CTG AGC AAA GCA GAC TAC GAG AAA CAC AAA GTC TAC GCC
`
`1037
`
`Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala-
`180
`185
`190
`
`TGC GAA GTC ACC CAT CAG GGC CTG AGC TCG CCC GTC ACA AAG AGC TTC
`
`1085
`
`Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
`195
`200
`205
`
`AAC AGG GGA GAG TGT T AAGCTGATCC TCTACGCCGG ACGCATCGTG
`
`Asn Arg Gly Glu Cys O C*
`210
`
`
`
`GCCCTAGTAC GCAACTAGTC GTAAAAAGGG TATCTAGAGG TTGAGGTGAT TTT ATG
`Met
`—23
`
`1131
`
`'
`
`1187
`
`BEGIN stII SIGNAL SEQUENCE
`AAA AAG AAT ATC GCA TTT CTT CTT GCA TCT ATG TTC GTT TTT TCT ATT
`
`1235
`
`Lys Lys Asn Ile Ala Phe Leu Leu Ala Ser Met Phe Val Phe Ser Ile
`-20
`-15
`—1O
`
`GCT ACA AAC GCG TAC GCT GAG GTT CAG CTG GTG GAG TCT GGC GGT GGC
`
`1283
`
`Ala Thr Asn Ala Tyr Ala Glu Val Gln Leu Val Glu Ser Gly Gly Gly
`—5
`1
`5
`10
`
`CTG GTG CAG CCA GGG GGC TCA CTC CGT TTG TCC TGT GCA GCT TCT GGC
`
`1331
`
`Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
`15
`20
`25
`BEGIN HEAVY CHAIN
`
`TAT ACC TTC ACC AAC TAT GGT ATG AAC TGG ATC CGT CAG GCC CCG GGT
`Tyr Thr‘Phe Thr Asn Tyr Gly Met Asn Trp Ile Arg Gln Ala Pro Gly
`30
`35
`4O
`'
`
`1379
`
`AAG GGC CTG GAA TGG GTT GGA TGG ATT AAC ACC TAT ACC GGT GAA CCG
`
`1427
`
`Lys Gly Leu Glu Trp Val Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro
`45
`50
`55
`
`ACC TAT GCT GCG GAT TTC AAA CGT CGT TTT ACT ATA TCT GCA GAC ACC
`Thr Tyr Ala Ala Asp Phe Lys Arg Arg Phe Thr Ile Ser Ala Asp Thr
`60
`65
`70
`
`1475
`
`TCC AGC AAC ACA GTT TAC CTG CAG ATG AAC AGC CTG CGC GCT GAG GAC
`
`1523
`
`Ser Ser Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
`75
`8O
`85
`90
`
`ACT GCC GTC TAT TAC TGT GCA AAG TAC CCG CAC TAT TAT GGG AGC AGC
`
`1571
`
`Thr Ala Val Tyr Tyr Cys Ala Lys Tyr Pro His Tyr Tyr Gly Ser Ser
`95
`100
`105
`
`'”
`
`FIG._ 8B
`
`a. I_~—_I .—— -. ..___ ._- a. — an--
`
`Regeneron Exhibit 1024.0814
`
`

`

`1'
`
`W0 98/4533]
`
`' PCTIUS98/06604
`
`1
`1
`
`1??
`
`.
`g
`
`3,
`1
`sf
`
`CAC TGG TAT TTC GAC GTC TGG GGT CAA GGA ACC CTG GTC ACC GTC TCC
`His Trp Tyr Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser
`110
`115
`120
`
`9/16
`
`TCG GCC TCC ACC AAG GGC CCA TCG GTC TTC CCC CTG GCA CCC TCC TCC
`Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
`125
`130
`135
`
`AAG AGC ACC TCT GGG GGC ACA GCG GCC CTG GGC TGC CTG GTC AAG GAC
`Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
`140
`145
`V
`150
`
`TAC TTC CCC GAA CCG GTG ACG GTG TCG TGG AAC TCA GGC GCC CTG ACC
`Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
`155
`160
`165
`170
`
`AGC GGC GTG CAC ACC TTC CCG GCT GTC'CTA CAG TCC TCA GGA CTC TAC
`Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
`175
`180
`185
`
`TCC CTC AGC AGC GTG GTG ACC GTG CCC TCC AGC AGC TTG GGC ACC CAG
`Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln'
`»
`190
`195
`200
`
`ACC TAC ATC TGC AAC GTG AAT CAC AAG CCC AGC AAC ACC AAG GTC GAC
`Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
`205
`210
`215
`
`AAG AAA GTT GAG CCC AAA TCT TGT GAC AAA ACT CAC CTC TAG AGT GGC
`Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Leu
`* Ser Gly
`220
`225
`230
`
`GGT GGC TCT GGT TCC GGT GAT TTT GAT TAT GAA AAG ATG GCA AAC GCT
`Gly Gly Ser Gly Ser Gly Asp Phe Asp Tyr Glu Lys Met Ala Asn Ala
`235
`240
`245
`250
`END LIGHT CHAIN
`BE
`AAT AAG GGG GCT ATG ACC GAA AAT GCC GAT GAA AAC GCG CTA CAG TCT
`Asn Lys Gly Ala Met Thr Glu Asn Ala Asp Glu Asn Ala Leu Gln Ser
`255
`260
`‘ 265
`'
`
`GIN g3p DOMAIN
`GAC GCT AAA GGC AAA CTT GAT TCT GTC GCT ACT GAT TAC GGT GCT GCT
`Asp Ala Lys Gly Lys Leu Asp Ser Val Ala Thr Asp Tyr Gly Ala Ala
`270
`»
`275
`280
`
`ATC GAT GGT TTC ATT GGT GAC GTT TCC GGC CTT GCT AAT GGT AAT GGT
`Ile Asp Gly Phe Ile Gly Asp Val Ser Gly Leu Ala Asn Gly Asn Gly
`M» ~~~~~~285»7-15,7 -,_
`7220
`,
`295_
`
`1619
`
`1667
`
`1715
`
`1763
`
`1811
`
`1859
`
`1907
`
`1955
`
`2003
`
`2051
`
`2099
`
`2147
`
`GCT ACT GGT GAT TTT GCT GGC TCT AAT TCC CAA ATG GCT CAA GTCV GGTW ’ ”2195”
`Ala Thr Gly Asp Phe Ala Gly Ser Asn Ser Gln Met Ala Gln Val Gly
`300
`305
`310
`
`CAG GGT GAT AAT TCA CCT TTA ATG AAT AAT TTC CGT CAA TAT TTA CCT
`Asp Gly Asp Asn Ser Pro Leu Met Asn Asn Phe Arg Gln Tyr Leu Pro
`315
`'
`320
`325
`330
`
`2243
`'
`
`FIG._8C
`
`‘
`
`1
`
`\
`
`Regeneron Exhibit 1024.0815
`
`

`

`W0 98/4533]
`
`PCT/US98/06604
`
`TCC
`Ser
`
`CTC CCT CAA
`Leu Pro Gln
`
`TCG GTT GAA
`Ser Val Glu
`
`335‘
`
`10/16
`
`TGT CGC CCT
`
`Cys
`
`Arg Pro
`340
`
`AAA
`
`CCA TAT GAA
`
`Lys
`
`Pro Tyr Glu
`350
`
`TTT TCT ATT
`Phe Ser Ile
`
`GAT TGT GAC
`
`Asp
`
`Cys Asp
`355
`
`GGT
`
`Gly
`
`GTC TTT GCG
`Val Phe Ala
`365
`
`TTT CTT TTA TAT
`Phe Leu Leu
`
`GTT GCC
`
`Tyr Val Ala
`370
`
`ACG TTT GCT
`
`AAC ATA CTG
`
`CGT AAT AAG
`
`Asn Lys
`
`TTT
`
`Phe
`
`AAA
`
`Lys
`
`ACC
`Thr
`
`GAG
`Glu
`
`GTC TTT AGC GCT GGT
`
`2291
`
`Val Phe Ser Ala Gly
`345
`
`ATA AAC TTA TTC CGT
`
`2339
`
`Ile Asn Leu Phe Arg
`360
`
`TTT ATG TAT GTA TTT
`
`2387
`
`Phe Met Tyr Val Phe
`375
`
`TCT TAATCATGCC
`
`2433
`
`Ser OC* (SEQ.ID NO:100)
`390
`
`CCCGCGTTGC
`
`GTCGCGGTGC
`
`2493
`
`ACCTCGCTAA
`
`CGGATTCACC
`
`2553
`
`TCT
`Ser
`
`Thr Phe Ala Asn Ile Leu Arg
`380
`385
`AGTTCTTTTG
`
`ATGGAGCCGG
`
`GCTAGCGCCG CCCTATACCT TGTCTGCCTC
`END G3 PROTEIN
`GCCACCTCGA
`AGCCGGCGGC
`
`CCTGAATGGA
`
`ACTCCAAGAA
`
`TTGGAGCCAA
`
`TCAATTCTTG
`
`CGGAGAACTG
`
`TGAATGCGCA
`
`AACCAACCCT
`
`2613
`
`TGGCAGAACA
`
`TATCCATCGC
`
`GTCCGCCATC
`
`TCCAGCAGCC
`
`GCACGCGGCG
`
`CATCTCGGGC
`
`2573
`
`AGCGTTGGGT
`
`CCTGGCCACG
`
`GGTGCGCATG
`
`ATCGTGCTCC
`
`TGTCGTTGAG
`
`GACCCGGCTA
`
`2733
`
`GGCTGGCGGG
`
`GTTGCCTTAC
`
`TGGTTAGCAG
`
`AATGAATCAC
`
`CGATACGCGA
`
`GCGAACGTGA
`
`2793
`
`AGCGACTGCT
`
`GCTGCAAAAC
`
`GTCTGCGACC
`
`TGAGCAACAA
`
`CATGAATGGT
`
`CTTCGGTTTC
`
`2853
`
`GCACCATTAT
`
`CGTGTTTCGT
`
`AAAGTCTGGA
`
`AACGCGGAAG
`
`TCAGCGCCCT
`
`GTTCCGGATC
`
`2913
`
`TGCATCGCAG
`
`GATGCTGCTG
`
`GCTACCCTGT
`
`GGAACACCTA
`
`CATCTGTATT
`
`AACGAAGCGC
`
`2973
`
`TGGCATTGAC
`
`CCTGAGTGAT
`
`TTTTCTCTGG
`
`TCCCGCCGCA
`
`TCCATACCGC
`
`CAGTTGTTTA
`
`3033
`
`CCCTCACAAC
`
`GTTCCAGTAA
`
`CCGGGCATGT
`
`TCATCATCAG
`
`TAACCCGTAT
`
`CGTGAGCATC
`
`3093
`
`CTCTCTCGTT
`
`TCATCGGTAT
`
`CATTACCCCC
`
`ATGAACAGAA
`
`ATTCCCCCTT
`
`ACACGGAGGC
`
`3153
`
`ATCAAGTGAC
`
`CAAACAGGAA
`
`AAAACCGCCC
`
`TTAACATGGC
`
`CCGCTTTATC
`
`AGAAGCCAGA
`
`3213
`
`CATTAACGCT
`
`TCTGGAGAAA
`
`CTCAACGAGC
`
`TGGACGCGGA
`
`TGAACAGGCA
`
`GACATCTGTG
`
`3273
`
`AATCGCTTCA
`
`CGACCACGCT
`
`GATGAGCTTT
`
`ACCGCAGGAT
`
`CCGGAAATTG
`
`TAAACGTTAA
`
`3333
`
`TATTTTGTTA
`
`AAATTCGCGT
`
`TAAATTTTTG
`
`TTAAATCAGC
`
`TCATTTTTTA
`
`ACCAATAGGC
`
`3393
`
`CGAAATCGGC
`
`AAAATCCCTT
`
`ATAAATCAAA
`
`AGAATAGACC
`
`GAGATAGGGT
`
`TGAGTGTTGT
`
`3453
`
`TCCAGTTTGG
`
`AACAAGAGTC
`
`CACTATTAAA
`
`GAACGTGGAC
`
`TCCAACGTCA
`
`AAGGGCGAAA
`
`3513
`
`AACCGTCTAT
`
`CAGGGCTATG
`
`GCCCACTACG
`
`TGAACCATCA
`
`CCCTAATCAA
`
`GTTTTTTGGG
`
`3573
`
`GTCGAGGTGC
`
`CGTAAAGCAC
`
`TAAATCGGAA
`
`CCCTAAAGGG
`
`AGCCCCCGAT
`
`TTAGAGCTTG
`
`3633
`
`FIG._8D
`
`--._‘_._- .__ _. .——— a—II. — a.-.
`
`Regeneron Exhibit 1024.0816
`
`

`

`W0 98/4533!
`
`PCT/US98/06604
`
`ACGGGGAAAG CCGGCGAACG
`
`11 / 16
`TGGCGAGAAA
`GGAAGGGAAG
`
`AAAGCGAAAG
`
`GAGCGGGCGC
`
`3693
`
`TAGGGCGCTG.GCAAGTGTAG
`
`CGGTCACGCT
`
`GCGCGTAACC
`
`ACCACACCCG
`
`CCGCGCTTAA
`
`3753
`
`TGCGCCGCTA CAGGGCGCGT
`
`CCGGATCCTG
`
`CCTCGCGCGT
`
`TTCGGTGATG
`
`ACGGTGAAAA
`
`3813
`
`CCTCTGACAC ATGCAGCTCC
`
`CGGAGACGGT
`
`CACAGCTTGT
`
`CTGTAAGCGG
`
`ATGCCGGGAG
`
`3873
`
`CAGACAAGCC CGTCAGGGCG
`
`CGTCAGCGGG
`
`TGTTGGCGGG
`
`TGTCGGGGCG
`
`CAGCCATGAC
`
`3933
`
`CCAGTCACGT AGCGATAGCG
`
`GAGTGTATAC
`
`TGGCTTAACT
`
`ATGCGGCATC
`
`AGAGCAGATT
`
`3993
`
`GTACTGAGAG TGCACCATAT
`
`GCGGTGTGAA
`
`ATACCGCACA
`
`GATGCGTAAG
`
`GAGAAAATAC
`
`4053
`
`CGCATCAGGC GCTCTTCCGC
`
`TTCCTCGCTC
`
`ACTGACTCGC
`
`TGCGCTCGGT
`
`CGTTCGGCTG
`
`4113
`
`CGGCGAGCGG TATCAGCTCA
`
`CTCAAAGGCG
`
`GTAATACGGT
`
`TATCCACAGA
`
`ATCAGGGGAT
`
`4173
`
`AACGCAGGAA AGAACATGTG
`
`AGCAAAAGGC
`
`CAGCAAAAGG
`
`CCAGGAACCG
`
`TAAAAAGGCC
`
`4233
`
`GCGTTGCTGG CGTTTTTCCA
`
`TAGGCTCCGC
`
`CCCCCTGACG
`
`AGCATCACAA
`
`AAATCGACGC
`
`4293
`
`TCAAGTCAGA GGTGGéGAAA
`
`CCCGACAGGA
`
`CTATAAAGAT
`
`ACCAGGCGTT
`
`TCCCCCTGGA
`
`4353
`
`AGCTCCCTCG TGCGCTCTCC
`
`TGTTCCGACC
`
`CTGCCGCTTA
`
`CCGGATACCT
`
`GTCCGCCTTT
`
`4413
`
`CTCCCTTCGG GAAGCGTGGC
`
`GCTTTCTCAT
`
`AGCTCACGCT
`
`4473
`
`GTAGGTATCT
`
`CAGTTCGGTG
`
`TAGGTCGTTC GCTCCAAGCT
`
`GGGCTGTGTG‘
`
`CACGAACCCC
`
`CCGTTCAGCC
`
`CGACCGCTGC
`
`4533
`
`GCCTTATCCG GTAACTATCG
`
`TCTTGAGTCC
`
`AACCCGGTAA
`
`GACACGACTT
`
`ATCGCCACTG
`
`4593
`
`GCAGCAGCCA CTGGTAACAG
`
`GATTAGCAGA
`
`GCGAGGTATG
`
`TAGGCGGTGC
`
`TACAGAGTTC
`
`4653
`
`TTGAAGTGGT GGCCTAACTA
`
`CGGCTACACT
`
`AGAAGGACAG
`
`TATTTGGTAT
`
`CTGCGCTCTG
`
`4713
`
`CTGAAGCCAG TTACCTTCGG
`
`AAAAAGAGTT
`
`GGTAGCTCTT
`
`GATCCGGCAA
`
`ACAAACCACC
`
`4773
`
`GCTGGTAGCG GTGGTTTTTT
`
`TGTTTGCAAG
`
`CAGCAGATTA
`
`CGCGCAGAAA
`
`AAAAGGATCT
`
`4833
`
`CAAGAAGATC CTTTGATCTT
`
`TTCTACGGGG
`
`TCTGACGCTC
`
`AGTGGAACGA
`
`AAACTCACGT
`
`4893
`
`TAAGGGATTT TGGTCATGAG
`
`ATTATCAAAA
`
`AGGATCTTCA
`
`CCTAGATCCT
`
`TTTAAATTAA
`
`4953
`
`AAATGAAGTT TTAAATCAAT
`
`CTAAAGTATA
`
`TATGAGTAAA
`
`CTTGGTCTGA
`
`CAGTTACCAA
`
`5013
`
`”TGCTTAATCAwGTGAGGCACCL
`
`TATCTCAGCG.
`
`ATCTQTCTAT
`
`ATTCGZFTCFPC
`
`VCATAGTTGCC
`
`5073
`
`TGACTCCCCG TCGTGTAGAT
`
`AACTACGATA
`
`CGGGAGGGCT
`
`TACCATCTGG
`
`CCCCAGTGCT
`
`5133
`
`GCAATGATAC CGCGAGACCC
`
`ACGCTCACCG
`
`GCTCCAGATT
`
`TATCAGCAAT
`
`AAACCAGCCA
`
`5193
`
`GCCGGAAGGG CCGAGCGCAG
`
`AAGTGGTCCT
`
`GCAACTTTAT
`
`CCGCCTCCAT
`
`CCAGTCTATT
`
`5253
`
`FflGa=8E
`
`___.-. ._... — —_.
`
`Regeneron Exhibit 1024.0817
`
`

`

`WO 98/45331
`
`PC17US9&m6604
`
`12/16
`
`AATTGTTGCC'GGGAAGCTAG
`
`AGTAAGTAGT
`
`TCGCCAGTTA
`
`ATAGTTTGCG
`
`CAACGTTGTT
`
`5313
`
`GCCATTGCTG
`
`CAGGCATCGT
`
`GGTGTCACGC
`
`TCGTCGTTTG
`
`GTATGGCTTC
`
`ATTCAGCTCC
`
`5373
`
`GGTTCCCAAC
`
`GATCAAGGCG
`
`AGTTACATGA
`
`TCCCCCATGT
`
`TGTGCAAAAA
`
`AGCGGTTAGC
`
`5433
`
`TCCTTCGGTC
`
`CTCCGATCGT
`
`>TGTCAGAAGT
`
`AAGTTGGCCG
`
`CAGTGTTATC
`
`ACTCATGGTT
`
`5493
`
`ATGGCAGCAC
`
`TGCATAATTC
`
`TCTTACTGTC
`
`ATGCCATCCG
`
`TAAGATGCTT
`
`TTCTGTGACT
`
`GGTGAGTACT
`
`CAACCAAGTC
`
`ATTCTGAGAA
`
`TAGTGTATGC
`
`GGCGACCGAG
`
`TTGCTCTTGC
`
`5553
`
`5613
`
`CCGGCGTCAA
`
`CACGGGATAA
`
`TACCGCGCCA
`
`CATAGCAGAA
`
`CTTTAAAAGT
`
`GCTCATCATT
`
`5673
`
`GGAAAACGTT
`
`CTTCGGGGCG
`
`AAAACTCTCA
`
`AGGATCTTAC
`
`CGCTGTTGAG
`
`ATCCAGTTCG
`
`5733
`
`ATGTAACCCA
`
`CTCGTGCACC
`
`CAACTGATCT
`
`TCAGCATCTT
`
`TTACTTTCAC
`
`CAGCGTTTCT
`
`5793
`
`GGGTGAGCAA
`
`AAACAGGAAG
`
`GCAAAATGCC
`
`GCAAAAAAGG
`
`GAATAAGGGC
`
`GACACGGAAA
`
`5853
`
`TGTTGAATAC
`
`TCATACTCTT
`
`CCTTTTTCAA
`
`TATTATTGAA
`
`GCATTTATCA
`
`GGGTTATTGT
`
`5913
`
`CTCATGAGCG
`
`GATACATATT
`
`TGAATGTATT
`
`TAGAAAAATA
`
`AACAAATAGG‘
`
`GGTTCCGCGC
`
`5973
`
`ACATTTCCCC
`
`GAAAAGTGCC
`
`ACCTGACGTC
`
`TAAGAAACCA
`
`TTATTATCAT
`
`GACATTAACC
`
`6033
`
`TATAAAAATA
`
`GGCGTATCAC
`
`GAGGCCCTTT
`
`CGTCTTCAA
`
`6072
`
`FIG._8F
`
`
`
`
`
`(\- I—nm .—— an- -
`
`_—— 4—1-0 '- na‘
`
`Regeneron Exhibit 1024.0818
`
`

`

`W0 98/4533]
`
`PL'I'IU 598/0661”
`
`13/16
`
`F(ab)-12
`MB 1.6
`H2305.6
`Y0101
`Y0192
`
`F(ab)-12
`MB 1 .6
`H2305.6
`Y0101
`Y0192
`
`F(ab)-12
`MB 1 .6
`H2305.6
`Y0101
`Y0192
`
`10
`
`DIQMTQSPSS
`
`20
`LSASVGDRVT
`
`DI
`DI
`DI
`DI
`
`QSPSS
`QSPSS
`QSPSS
`QSPSS
`
`LSASVGDRVT
`LSASVGDRVT
`LSASVGDRVT
`LSASVGDRVT
`
`50
`GKAPKVLIYF
`
`GKAPKELIYF
`GKAPKVLIYF
`GKAPKVLIYF
`GKAPKVLIYF
`
`60
`TSSLHSGVPS
`TSSLHSGVPS
`TSSLHSGVPS
`TSSLHSGVPS
`TSSLHSGVPS
`CDR-L2
`
`90
`
`100
`
`EDFATYYCQQ YSTVPWTFGQ
`EDFATYYCQQ YSTVPWTFGQ
`EDFATYYCQQ YSTVPWTFGQ
`EDFATYYCQQ YSTVPWTFGQ
`
`EDFATYYCQQ YSTVPWTFGQ
`CDR-L3
`
`3O
`
`40
`
`ITCSASQDIS NYLNWYQQKP
`ITCSASQDIS NYLNWYQQKP
`ITCSASQDIS NYLNWYQQKP
`ITCSASQDIS NYLNWYQQKP
`IT
`S NYLNWYQQKP
`CDR-Ll
`7O
`RFSGSGSGTD
`RFSGSGSGTD
`RFSGSGSGTD
`RFSGSGSGTD
`RFSGSGSGTD
`
`FTLTISSLQP
`
`EELTISSLQP
`
`LTISSLQP
`FTLTISSLQP
`FTLTISSLQP
`
`80
`
`110
`GTKVEIKRTV
`GTKVEIKRTV
`GTKVEIKRTV
`GTKVEIKRTV
`GTKVEIKRTV
`
`(SEQ.ID NO:8)
`(SEQ.ID N02101)
`(SEQ.ID NO:103)
`(SEQ.ID NO: 105)
`(SEQ.ID NO: 107)
`
`FIG._9A
`
`NYGMNWVRQA
`NYG
`QA
`NYG
`QA
`NYGMNWVRQA
`SCAASGYTFT NYGQEWVRQA
`CDR-H 1
`7o
`60
`AADFKRRFTF
`INTYTGEPTY
`INTYTGEPTY
`AADFKRRFTE
`AADFKRRFTF
`INTYTGEPTY
`AADFKRRFTF
`INTYTGEPTY
`INTYTGEPTY
`AADFKRRFTF
`CDR-HZ
`110
`100
`TAVYYCAKYP HYYGSSHWYF
`
`40
`
`SLDTSKSTAY
`
`$33?
`
`SLDTSKSTAY
`SLDTSKSTAY
`CDR-7
`
`DVWGQGTL
`DVWGQGTL
`DVWGQGTL
`DVWGQGTL
`DVWGQGTL
`
`(SEQ.ID N027)
`(SEQ.ID NO:102)
`(SEQ.IDNO: 104)
`(SEQ.ID N02106)
`(SEQ.ID NO: 108)
`
`F(ab)—12
`MB].6
`H2305.6
`Y0101
`Y0192
`
`F(ab)- 12
`MB 1 .6
`H2305.6
`Y0101
`Y0192
`
`F(ab)-12
`V ‘MBl.6’
`H2305.6
`Y0101
`Y0192
`
`10
`EVQLVESGGG
`EVQLVESGGG
`EVQLVESGGG
`EVQLVESGGG
`EVQLVESGGG
`
`SO
`PGKGLEWVGW
`PGKGLEWVGW
`PGKGLEWVGW
`PGKGLEWVGW
`PGKGLEWVGW
`
`90
`
`LQMNSLRAED
`LQMNSLRAEDV
`LQMNSLRAED
`LQMNSLRAED
`LQMNSLRAED
`
`20
`
`LVQPGGSLRL
`LVQPGGSLRL
`LVQPGGSLRL
`LVQPGGSLRL
`LVQPGGSLRL
`
`30
`SCAASGYTFT
`SCAASGYTFT
`SCAASGYTFT
`SCAASGYTFT
`
`~TAVYYCAKYPVHXYQSSHWYF
`TAVYYCAKYP HYYGSSHWYF‘
`TAVYYCAKYP HYYGSSHWYF
`TAVYYCAKYP HYYGSSHWYF
`CDR—H3
`
`'FIG._QB
`
`as. .— --.—. .— -n u-.— l.‘ n
`
`Regeneron Exhibit 1024.0819
`
`
`
`
`
`..1..-1“._.__._._.._...
`
`,
`
`
`
`
`
`.._”III-M__.._.m_.t_._..1.._
`
`

`

`wo 98/453251
`
`PCT/US98/06604
`
`14/ 16
`
`10
`
`20
`
`30
`
`4o
`
`DIQMTQSPSS LSASVGDRVT ITCSASQDIS NYLNWYQQKP
`
`QSPSS LSASVGDRVT mm
`DI
`s NYLNWYQQKP
`
`QSPSS LSASVGDRVT ITO ‘
`DI
`s NYLNWYQQKP
`DI
`QSPSS LSASVGDRVT ITO"
`NYLNWYQQKP
`DI
`QSPSS LSASVGDRVT ITCSASQDIS NYLNWYQQKP
`CDR-L1
`50
`60
`7o
`80
`GKAPKVLIYF TSSLHSGVPS RFSGSGSGTD FTLTISSLQP
`GKAPKVLIYF TSSLHSGVPS RFSGSGSGTD FTLTISSLQP
`GKAPKVLIYF TSSLHSGVPS RFSGSGSGTD FTLTISSLQP
`GKAPKVLIYF TSSLHSGVPS RFSGSGSGTD FTLTISSLQP
`GKAPKVLIYF TSSLHSGVPS RFSGSGSGTD FTLTISSLQP
`CDR-L2
`
`90
`
`100
`
`110
`
`EDFATYYCQQ YSTVPWTFGQ GTKVEIKRTV .
`EDFATYYCQQ YSTVPWTFGQ GTKVEIKRTV
`EDFATYYCQQ YSTVPWTFGQ GTKVEIKRTV
`EDFATYYCQQ YSTVPWTFGQ GTKVEIKRTV
`EDFATYYCQQ YS'I'VPWTFGQ GTKVEIKRTV
`CDR-L3
`
`(SEQ.ID N028)
`(SEQ.ID NO:109)
`(SEQ.ID NO:111)
`(SEQ.ID NO:113)
`(SEQ.ID NO:115)
`
`F(ab)—12
`Y0243-1
`Y0238-3
`Y0313-1
`Y0317
`
`F(ab)-l2
`Y0243-l
`Y0238-3
`YO3l3-l
`Y0317 V
`‘
`
`F(ab)-12
`Y0243-1
`Y0238-3
`Y0313-1
`Y0317
`
`
`
`10
`
`20
`
`30
`
`40
`
`EVQLVESGGG LVQPGGSLRL SCAASGYTFT NYGMNWVRQA I
`EVQLVESGGG LVQPGGSLRL SCAASGYQFT @YGMNWVRQA
`EVQLVESGGG LVQPGGSLRL SCAASGYTFT NYGENWRQA
`EVQLVESGGG LVQPGGSLRL SCAASG
`GMNWVRQA
`EVQLVESGGG LVQPGGSLRL SCAASG
`GMNWVRQA
`
`50
`
`60
`
`80
`
`T
`CDR-Hl
`7o
`
`F(ab)-12
`Y0243-1
`Y0238-3
`Y03l3-1
`Y0317
`
`F(ab)-12
`Y0243-l
`Y0238-3
`-. Y0313—1
`Y0317
`
`PGKGLEWVGW INTYTGEPTY AADFKRRFTF SLDTSKSTAY
`_PGKGLEWVGW INTYTGEPTY AADFKRRFTF SLDTSKSTAY
`PGKGLEWVGW INTYTGEPTY AADFKRRFTF SLDTSKSTAY
`PGKGLEWVGW INTYTGEPTY AADFKRRFTF SLDTSKSTAY
`PGKGLEWVGW INTYTGEPTY AADFKRRFTF SLDTSKSTAY
`CDR-H2
`CDR-7
`100
`
`110
`
`90
`
`F(ab)-l2
`Y0243—1
`Y0238-3
`YO313-l
`Y0317
`
`LQMNSLRAED TAVYYCAKYP HYYGSSHWYF DVWGQGTL
`LQMNSLRAED TAVYYCAKYP HYYGSSHWYF DVWGQGTL
`LQMNSLRAED TAVYYCAKYP
`HWYF DVWGQGTL
`LQMNSLRAED TAVYYCAKYP
`HWYF DVWGQGTL
`LQMNSLRAED TAVYYCAKYP
`SHWYF DVWGQGTL
`CDR-H3
`
`(SEQ.ID N027)
`(SEQ.ID NO:110)
`(SEQ.ID NO:112)
`(SEQ.ID NO:114)
`(SEQ.ID NO:116)
`
`FIG._ 103
`
`Regeneron Exhibit 1024.0820
`
`

`

`
`
`._.V._.~.;._u:_-..._.._.__________._.__..._4_”—__.__4_~-._.
`
`
`
`,._____..'_.__._‘.L_'-:__....
`
`WO'98/45331
`
`PCTIUS98/06604
`
`15/16
`
`
`
`
`
`[3H]—THYMIDINEINCORPORATION
`
`3000
`
`2500
`
`'2000
`
`1500
`
`1000
`
`500
`
`EQ.
`3
`
`
`
`Y0238-3
`- Y0192
`F(ab) —12
`Y0313-1
`
`--
`
`0.001
`
`0.01
`
`0.1
`
`1
`
`10
`
`100
`
`[ANTIBODY], nM
`
`FIG._ 11
`
`A. .—__._- -_._ -. ..__—_ -—. ..
`
`Regeneron Exhibit 1024.0821
`
`

`

`W0 98/4533]
`
`PCTIU898/06604
`
`Mm
`
`88m_6‘Hjug\wo<
`82OS9.Fto.5.0Sod
`
`
`
`
`8mm3<2.>:z_&$mom;DEE.-o--
`
`"am;$22.I#1
`
`
`
`“18>9&2:Iol
`
`oooom
`
`oooom
`
`88v___m
`
`:259:2moem“.
`
`oooom
`
`.0009NW[.mhm
`
`Regeneron Exhibit 1024.0822
`
`

`

`z
`
`:7
`
`in»,L.“‘a
`
`
`
`
`
`5;:.\._.L?r_‘:-filfihmwrlfiii
`‘ua»...z,uMm.
`
`
`
`
`
`Regeneron Exhibit 1024.0823
`
`
`
`
`
`
`
` .....i...x.1..V:.ArhonWAJ‘IIIIiI.islallltunlv;$.Jliflduzsii‘lal,
`
`
`

`

`PCT
`
`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`International Bureau
`
`
`
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(51) International Patent Classification 6 :
`C07K 16/22, C12N 15/13, 15/63,
`15/70, A61K 39/395
`
`_
`
`(11) International Publication Number:
`
`WO 98/45331
`
`(43) International Publication Date:
`
`15 October 1998 (15.10.98)
`
`(81) Designated States: AL, AM, AT, AU, AZ, BA, BB, BG, BR,
`BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE,
`GH, GM, GW, HU, ID, IL, IS, JP, KE, KG, KP, KR, [(2,
`LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW,
`MX, NO, NZ, PL, PT, RO, RU, SD, SE, 86, SI, SK, SL,
`TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZW, ARIPO
`patent (GH, GM, KE, LS, MW, SD, 82, UG, ZW), Eurasian
`patent (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), European
`patent (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR,
`IE, IT, LU, MC, NL, PT, SE), OAPI patent (BF, BJ, CF,
`CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG).
`
`Published
`With international search report.
`
`(88) Date of publication of the international search report:
`’
`3 December 1998 (03.12.98)
`
`
`
`Ia«L......~-.._.._._....._-_..
`
`(21) International Application Number:
`
`PCT/US98/06604
`
`(22) International Filing Date:
`
`3 April 1998 (03.04.98)
`
`(30) Priority Data:
`08/833,504
`08/908,469
`
`7 April 1997 (07.04.97)
`6 August 1997 (06.08.97)
`
`US
`US
`
`(71) Applicant (for all designated States except US): GENENTECH,
`INC. [US/US]; One DNA Way, South San Francisco, CA
`94080 (US).
`
`(72) Inventors; and
`(7S) Inventors/Applicants (for US only): BACA, Manuel [AU/US];
`Apartment #HS, 888 Foster City Boulevard, Foster City, CA
`94404 (US). WELLS, James, A. [US/US]; 1341 Columbus
`Avenue, Burlingame, CA 94010 (US). PRESTA, Leonard,
`G.
`[US/US]; Apartment 206, 1900 Gough Street, San
`Francisco, CA 94109 (US); LOWMAN, Henry, B. [US/US];
`400 San Juan Avenue, El Granada, CA 94018 (US). CHEN,
`Yvonne, Man—Yee [CA/US]; 1951 O'Farrell Street #321,
`San Mateo, CA 94403 (US).
`
`(74) Agents: DREGER, Walter, H. et al.; Flehr, Hohbach, Test,
`Albritton & Herbert LLP, Suite 400, 4 Embarcadero Center,
`San Francisco, CA 94111—4187 (US).
`
`(54) Title: ANTI-VEGF ANTIBODIES
`
`(57) Abstract
`
`Humanized and variant anti—VEGF antibodies and vari-
`ous uses therefor are disclosed. The anti—VEGF antibodies have
`strong binding affinities for VEGF; inhibit VEGF—induced pro-
`liferation of endothelial cells in vitro, and inhibit tumor growth
`in VIVO.
`
`
`
`TRANSFORM E. coli
`
`+M13KO7HELPERPHAGEl
`
`FAB-pm FUSION
`""‘\
`
`Regeneron Exhibit 1024.0824
`
`

`

`FOR THE PURPOSES OF INFORMATION ONLY
`
`Codes used to identify States party to the PCI‘ on the front pages of pamphlets publishing international applications under the PCT.
`
`Zimbabwe
`
`ES
`FI
`FR
`GA
`GB
`GE
`GH
`GN
`GR
`HU
`JE,
`[L
`IS
`IT
`JP
`KE
`KG
`KP
`
`Albania
`Armenia
`Austria
`Australia
`Azerbaijan
`Bosnia and Herzegovina
`Barbados
`Belgium
`Burkina Faso
`Bulgaria
`' Benin
`Brazil
`Belams
`Canada
`Central African Republic
`Congo
`Switzerland
`C016 d'lvoir'e
`Cameroon
`. China
`Cuba
`Czech Republic
`Germany
`Denmark
`Estonia
`
`Spain
`Finland
`France
`Gabon
`United Kingdom
`Georgia
`Ghana
`Guinea
`Greece
`Hungary
`Inland, ,
`
`Democratic People's
`Republic of Korea
`Republic of Korea
`Kazakstan
`Saint Lucia
`Liechtenstein
`Sri Lanka
`Liberia
`
`LS
`LT
`LU
`LV
`MC
`MD
`MG
`MK
`
`ML
`MN
`MR
`MW
`MX
`NE
`NL
`NO
`NZ
`PL
`PT
`RO
`RU
`SD
`-SE
`SG
`
`Lesotho
`Lithuania
`lauembcurg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`The former Yugoslav
`Republic of Macedonia
`Mali
`Mongolia
`Mauritania
`Malawi
`Mexico
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Singapore
`
`SI
`SK
`SN
`
`Slovenia
`Slovakia
`Senegal
`Swaziland
`Chad
`Togo
`Tajikistan
`Turkmenistan
`Turkey
`Trinidad and Tobago
`Ukraine
`
`7 ,M
`rUgandak _
`United States of Americana. m
`Uzbekistan
`Viet Nam
`Yugoslavia
`
`Ii i
`
`'l
`l.
`-'l
`
`liI
`
`
`
`!i[
`
`
`
`
`
`?77
`l
`
`
`
`Regeneron Exhibit 1024.0825
`
`

`

`ATIONAL SEARCI’I REPORT .
`
`ional Application No
`lntt
`PCT/US 98/06604
`
`A. CLASSIFICATION OF S BJECT MATTER
`IPC 6
`C07Kl6 22
`C12N15/13
`
`C12N15/63
`
`ClZNlS/70
`
`A61K39/395
`
`According to lntemational Patent Classiticatiom'lPC) or to both national classification and l PC
`8. FIELDS SEARCHED
`Minimum documentation searched (classification system lollowed by classification symbol5)
`IPC 6
`C07K C12N A61K
`
`Documentation searched other than minimumdocumentation to the extent that such documents are included in the tields searched
`
`Electronic data base consulted during the international search (name or data base and, where practical. search terms used)
`
`C. DOCUMENTS CONSIDERED TO BE RELEVANT
`
`Citation ot document. with indication, where appropriate. of the relevant passages
`
`I
`
`
`
`X
`
`Y
`
`Y
`
`NO 92 22653 A (GENENTECH INC)
`23 December 1992
`
`the whole document and specially: see
`SEO.ID.N. 17,18 and 25
`see page 5,
`line 24 - page 7,
`
`line 35
`
`see page 9,
`figure 5
`
`line 22 - page 10,
`
`line 4;
`
`KIM ET AL.,:
`"Inhibition of vascular
`endothelial growth factor—induced
`angiogenesis suppresses tumor growth in
`vivo"
`
`NATURE,
`vol. 362, 1993, page 841 XP002013864
`London, GB
`c