1111111111111111 IIIIII IIIII 1111111111 11111 1111111111 111111111111111 11111 1111111111 11111111
`US 20150216970Al
`
`c19) United States
`c12) Patent Application Publication
`GROGAN et al.
`
`(54) METHODS OF TREATING CANCER USING
`PD-1 AXIS BINDING ANTAGONISTS AND
`TIGIT INHIBITORS
`
`(71) Applicant: GENENTECH, INC., South San
`Francisco, CA (US)
`
`(72)
`
`Inventors: Jane GROGAN, San Francisco, CA
`(US); Robert J. JOHNSTON, San
`Francisco, CA (US); Bryan IRVING,
`San Francisco, CA (US); Jason
`HACKNEY, San Carlos, CA (US); Xin
`YU, South San Francisco, CA (US); Dan
`EATON, San Rafael, CA (US); Kristin
`BOWLES, South San Francisco, CA
`(US); Laetitia COMPS-AGRAR, Foster
`City, CA (US)
`
`(21) Appl. No.: 14/333,375
`
`(22) Filed:
`
`Jul. 16, 2014
`
`Related U.S. Application Data
`
`(60) Provisional application No. 61/846,941, filed on Jul.
`16, 2013, provisional application No. 61/865,582,
`
`c10) Pub. No.: US 2015/0216970 Al
`Aug. 6, 2015
`(43) Pub. Date:
`
`filed on Aug. 13, 2013, provisional application No.
`61/950,754, filed on Mar. 10, 2014, provisional appli(cid:173)
`cation No. 61/985,884, filed on Apr. 29, 2014, provi(cid:173)
`sional application No. 61/992,109, filed on May 12,
`2014.
`
`Publication Classification
`
`(51)
`
`Int. Cl.
`A61K39/395
`A61K 45106
`(52) U.S. Cl.
`CPC ....... A61K 39/3955 (2013.01); A61K 39/39558
`(2013.01); A61K 45106 (2013.01)
`
`(2006.01)
`(2006.01)
`
`(57)
`
`ABSTRACT
`
`The present invention describes combination treatment com(cid:173)
`prising a PD-1 axis binding antagonist and an agent that
`decreases or inhibits TIGIT expression and/or activity and
`methods for use thereof, including methods of treating con(cid:173)
`ditions where enhanced immunogenicity is desired such as
`increasing tumor immunogenicity for the treatment of cancer
`or chronic infection.
`
`Merck Ex. 1005
`Page 1 of 140
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`

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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 1 of 65
`
`US 2015/0216970 Al
`
`D
`300
`250
`U: 200
`::l?;
`t- '150
`~ 100-
`j:: bO
`0-Lli-~~u.:.,...
`
`~t~~~-s~-t~
`,f:><:_,.o t>.'<:io ~
`,rp bl
`
`FIG.1A-1F
`
`A
`
`B
`
`C
`
`Hours
`
`Resting
`
`TIGIT
`
`TIGJT
`
`TIGIT
`
`Naive coa+
`---TEM co4+
`------ TEM coa+
`
`F
`100
`
`80
`+
`!:: 00·
`C)
`i= 40
`~ "'
`20
`
`E
`
`TIGIT
`
`Merck Ex. 1005
`Page 2 of 140
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`

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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 2 of 65
`
`US 2015/0216970 Al
`
`H
`2500·
`
`2000··
`
`i moo
`I(cid:173)S2 1000 ..
`500 ·
`
`~
`
`J
`2400
`
`1000
`
`FIG. 1G-1J
`
`G
`
`Naive COB+
`- -Po-1tow TEM
`------po.1Mgh TEM
`
`Naive
`TCM
`- - - TEM po.1 low
`---··- TEM Po-1nigh
`
`TiGIT
`
`Merck Ex. 1005
`Page 3 of 140
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`

`

`30
`
`40
`
`50
`
`60
`
`'\.
`
`\70 I
`
`/
`
`80
`
`90
`
`Frt-PGK-em7-NEO-FRT
`
`31 kb
`
`Tigit
`
`3'Extra
`for PCR
`
`3'Arm
`
`N!!r,.
`
`Floxed
`Region
`
`5'Arm
`
`3'Extra
`forPCR
`
`Total Length Homology Arms= 6030 bp
`5'Homology Arm = 2405 bp
`Floxed Region Contain Exon1 = 937 bp
`3'Homology Arm = 3175 bp
`Selection Marker= Frt- PGK-em7-NEO- FRT
`
`Annotations
`~ LINE ~ LTR ~ SNE
`-
`Exon
`c::=::=J UTR
`k.,., .. ,.,.;, CNS
`
`Control Vector
`Screening by PCR Followed by Sequencing
`5'Extra Genomic Region = 556 bp
`3'Extra Genomic Region = 284 bp
`
`, ... ., .. , Other ~ RNA
`
`FIG. 2
`
`"-= ~ .....
`
`(')
`
`~ .....
`
`(')
`
`~ .....
`
`('D = ..... t "e -....
`.... 0 =
`"-= = O" -....
`.... 0 =
`~
`~
`~Cl's
`N
`0 .....
`Ul
`
`('D
`('D
`
`rJ'1 =(cid:173)
`.....
`0 .....
`Cl's
`Ul
`
`~
`
`c
`
`rJ'1
`N
`0 .....
`Ul ---0
`N .....
`Cl's
`1,0
`-....J
`0
`
`> .....
`
`Merck Ex. 1005
`Page 4 of 140
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`

`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 4 of 65
`
`US 2015/0216970 Al
`
`FIG. 3A-3F
`
`A
`
`B
`
`C
`
`WT
`
`CKO
`
`D
`
`WT
`
`60%
`
`E
`
`CKO
`
`60%
`
`it!;:------
`CDB
`
`F
`
`U) 80
`Cl) u
`.,_ 60
`+ co 8 40
`+ ~20 ..
`LL
`'#- Q-&1....,.i...i...,..
`
`~&
`
`Merck Ex. 1005
`Page 5 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 5 of 65
`
`US 2015/0216970 Al
`
`FIG. 3G-3L
`
`G
`
`WT
`
`H
`
`CKO
`
`"tj"
`
`se
`
`CD4
`
`"'
`(1) 60
`<..>
`I-
`+ 40
`-.::i"
`C
`<..>
`"C 20
`(ti
`>
`.::;
`u
`
`0
`
`(1) -
`<( ~&
`
`~ 0
`
`J
`
`WT
`
`--- 12%
`
`K
`
`CKO
`
`L
`
`(1)
`
`.!!? 20
`~ 15 ..
`+
`~ 10
`<..>
`tn 5
`z
`LL 0
`~ c:> ~ &
`
`Merck Ex. 1005
`Page 6 of 140
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`

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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 6 of 65
`
`US 2015/0216970 Al
`
`FIG. 4A-4F
`
`A
`TIGITfl/fl CD4 WT
`
`B
`TIGITfl/fl CD4cre
`
`C
`
`,!12
`
`~45
`I-
`+
`~30
`u
`
`-g 15 -ffl >
`ti O -1L.a.;.;a.i...i......-L(cid:173)
`<C
`'#- ~&
`
`E
`TIGITfl/fl CD4 ere
`
`F
`
`Merck Ex. 1005
`Page 7 of 140
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`

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`Patent Application Publication
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`Aug. 6, 2015 Sheet 7 of 65
`
`US 2015/0216970 Al
`
`FIG. 4G-4L
`
`G
`TIGITfl/fl co4WT
`
`H
`TIGITfl/fl co4cre
`
`K
`TIGITfl/fl co4cre
`
`L
`
`** r1
`
`04
`Cl>
`~3
`+
`~2
`u
`+1
`en
`if: 0 _u,;.;,,;..J,-1.-,-L
`
`Merck Ex. 1005
`Page 8 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 8 of 65
`
`US 2015/0216970 Al
`
`FIG. 4M-4R
`
`M
`
`N
`
`0
`
`-g 100000
`:5
`LL e:,
`...
`.2!
`i=
`>
`::15:
`~
`a;
`>
`::::i
`
`10000
`
`1000 .........,_.....,...._
`-f cf
`
`Merck Ex. 1005
`Page 9 of 140
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`

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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 9 of 65
`
`US 2015/0216970 Al
`
`FIG. 4S-4X
`s
`
`T
`
`u
`
`Anti-PDL 1
`
`Anti-TIGIT
`
`V
`
`Anti-PDL1 +
`Anti-Tl GIT
`
`w
`+
`00
`C
`
`20-
`16
`
`<..) 1$1 -- 12
`
`o-a;
`+ <..) 8
`~ ....
`4-
`!:!::
`0---"+'-"+'-""""'"-.-'(cid:173)
`Anti-PDL 1 - + - +
`Anti-TIGIT -
`- + +
`
`X
`
`10000
`
`::::. 100000
`E
`:3
`I.I.. e:,
`-i=
`Q)
`> :a:
`
`1000 ..
`
`0
`...I
`Q)
`>
`100
`:.::i
`Anti-POL 1
`Anti-TIGIT
`
`*
`
`..
`..
`;l.~
`• ~
`
`1'
`
`,,
`-----------
`
`- +
`- +
`-
`- + +
`
`Merck Ex. 1005
`Page 10 of 140
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`

`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 10 of 65
`
`US 2015/0216970 Al
`
`FIG. 5A-5J
`
`A
`
`B
`
`C
`
`Control
`
`F
`
`G
`
`H
`
`Control
`
`Anti-POL 1
`
`8%~ w
`-tb ..
`·t:r/i
`-
`
`z- .
`
`LL
`
`CD4
`
`Anti-TIGIT
`
`Anti-PDL 1
`+ Anti-TIGIT
`
`E
`
`J
`
`+
`"3"
`C
`
`20
`16
`
`+U 8
`~I-
`4
`!:!::
`~ 0
`Anti-POL 1 - + - +
`Anti-Tl GIT - - + +
`
`01.11 --Oa, 12
`r:mlf:ol
`LlJLlJ
`
`Merck Ex. 1005
`Page 11 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 11 of 65
`
`US 2015/0216970 Al
`
`FIG. 6A-6C
`
`A
`
`5
`N"4
`C') g_3
`1-6 2-
`j:: 1 ··
`
`0
`
`B
`
`5--
`N°4-
`C) g_ 3
`!:: 2-
`(!)
`j:: 1.
`
`0 '
`
`C
`
`15
`
`1.0
`
`<,.)
`M
`C
`(.) 0..0
`
`-1 0
`
`~
`
`"Tumor
`• Normal
`
`ov ~
`
`<c, " ,,,,
`
`".::>
`TIGIT
`
`\),
`
`v.,
`
`Merck Ex. 1005
`Page 12 of 140
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`

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`Patent Application Publication
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`Aug. 6, 2015 Sheet 12 of 65
`
`US 2015/0216970 Al
`
`FIG. 6D-6H
`
`D
`
`5
`
`4
`
`-1 -
`
`,
`
`:,,"'':/;
`
`0 Tumor
`• Normal
`
`..
`
`E
`
`2
`
`v
`Q
`U-1---
`
`0
`
`, Tumor
`• Normal
`
`G
`
`"": 2
`
`~1
`
`0
`
`-1
`
`, Tumor
`• Normal
`
`"Tumor
`• Normal
`
`0'
`
`0
`
`0
`
`..
`
`.(cid:144)
`
`•
`
`""
`
`H
`
`2.0
`
`"'2' 1.5
`,q
`...J 1.0-
`I-
`U 0.5
`
`0.0
`
`Merck Ex. 1005
`Page 13 of 140
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`

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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 13 of 65
`
`US 2015/0216970 Al
`
`FIG. 7A-7D
`
`Spleen
`- - - - TIL PD-i low
`------ TIL PD-1 high
`
`A
`
`TIGIT
`
`TIGIT
`
`**
`
`B
`
`900---
`
`u:::600
`:E
`i-
`5300---
`.::
`
`***
`
`D
`
`1200-(cid:173)
`
`u:::
`·::e 800---
`1-0 400---
`.:::
`
`Merck Ex. 1005
`Page 14 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 14 of 65
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`US 2015/0216970 Al
`
`FIG. 7E-7F
`
`E
`
`10000
`
`o Control
`0 aPD-U
`4 uTIGIT
`v rxPD-LHnTIGIT
`
`:,g:_v
`""-
`--------~~~-...;;~ .... · -,;;;...--1,~· --...;
`
`~.~-m_p_le_te_,_R_oo_p_o-n_se ___ (C_R_) ...... ~/-r
`20
`0
`10
`30
`Day
`
`60
`
`10
`
`FIG.
`
`F
`
`100-t---~
`
`o Control
`(cid:143) a.PD-L 1
`"a.TIGIT
`v a.PD-L 1 + aTIGIT
`
`"iii
`-~
`> 80
`~ 60 -;
`
`40--
`
`i-!
`&_ 20
`0 .......,.-,--.,..,...,-0,-.,...,..,-,a-......,...,..,...,..,......,..,..,..,...,...,._...,
`20
`40
`60
`0
`
`Day
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 15 of 65
`
`US 2015/0216970 Al
`
`FIG. 7G-7K
`
`H
`
`10000;
`
`1000
`
`100.
`
`10
`
`0
`
`5
`
`10
`
`i5 20
`
`25
`
`K
`
`10001) ·.·
`
`10
`
`15
`
`20
`
`25
`
`Day
`
`J
`
`10000
`
`10000 •
`
`1000.
`
`100.
`
`10 - - - - - . - - - - - -
`0
`5
`10
`15
`20
`25
`
`10 '""", - - - - - , - - - - - ,
`0
`5
`10
`15
`20
`25
`
`10 ------,----,---,
`0
`5
`10
`15
`20
`25
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 16 of 65
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`US 2015/0216970 Al
`
`FIG. 7L-7P
`L
`
`Contra!
`
`M
`
`anti-POL 1
`
`N
`
`anti-TIGIT
`
`0
`
`anti-POL 1
`+ anti-TIGIT
`
`11%
`
`~-
`
`~ u~~~"!'M'IT"!"
`
`--;
`
`CD8
`
`p
`
`45 .,,
`
`*** n
`
`Merck Ex. 1005
`Page 17 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 17 of 65
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`US 2015/0216970 Al
`
`FIG. SA-8D
`
`A
`
`Spleen
`- - - - TIL Tim-31ow
`--·-·- TIL Tim-3high
`cos+
`
`TIGIT
`
`B
`
`900-
`
`§iooo-
`1-
`(5300··
`i=
`
`C
`
`Spleen
`----TIL T1m-31ow
`------ TIL T1m-3high
`u:
`co4+
`:!
`I-
`
`C .::
`
`TIGIT
`
`***
`
`D
`
`1500··-
`
`iI: 2;1000-
`!::
`C) 500 ...
`i=
`
`Merck Ex. 1005
`Page 18 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 18 of 65
`
`US 2015/0216970 Al
`
`B
`
`000--(cid:173)
`
`iBoo--(cid:173)
`C:
`(!)300···
`j::
`
`Spleen
`----- TIL PD-1/Tirn-3I~w
`---- Ttl PD~1mrn-3high
`coa+
`
`;,
`
`'
`)·-._
`,,,.
`
`l
`~
`\
`
`TIGIT
`
`-
`
`- -
`
`--
`
`FIG. 9A-9D
`
`A
`
`C
`
`Spieen
`-- - TIL PD-1/Tim-31~w
`--- TIL PD-1/Tim-Jhigh
`cn4+
`
`D
`
`900--(cid:173)
`
`i 600··
`!::
`(!) 300···
`~
`
`**
`
`TIGIT
`
`Merck Ex. 1005
`Page 19 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 19 of 65
`
`US 2015/0216970 Al
`
`Control
`
`0
`
`10
`
`30
`
`20
`Day
`
`h
`40 60
`
`10000
`
`' - -
`OM
`EE
`~s
`c: Cl) 1000
`.!!! E
`"t, ::I
`Cl)-
`:E~
`
`100
`
`10000
`
`Anti-PD-L 1
`
`0
`
`10
`
`30
`
`20
`Day
`
`h
`40 60
`
`10000
`
`' - -
`OM
`EE
`:::::i E
`1--
`c: Cl) 1000
`.!!! E
`"t, ::I
`Cl)-
`~~
`
`100
`
`10000
`
`' - -
`OM
`~ ~ 1000
`1--
`C: Cl)
`.!!! E
`"t, ::I 100
`Cl)-
`~~
`
`' - -
`OM
`~ ~ 1000
`Anti-TIGIT 1--
`C: Cl)
`.!!! E
`"t, ::I 100 .
`Cl)-
`~~
`
`Anti-TIGIT +
`Anti-PD-L 1
`
`.. ,, -----El
`
`h
`40 60
`
`10
`
`0
`
`10
`
`30
`
`20
`Day
`
`h
`40 60
`
`10
`
`0
`
`10
`
`20
`Day
`
`30
`
`FIG.10
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 20 of 65
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`US 2015/0216970 Al
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`FIG. 11A-11 C
`
`A
`
`45
`
`ti)
`...J
`j:::
`+ 30
`'q'
`C u
`t, 15
`z
`LI..
`
`B
`
`C
`
`20
`(/).t:.
`- C.
`-; E
`u~ 15-
`I- Cl
`+ c:-
`'q' ·- (U
`c.E-g 10
`UEz
`+"C
`O'>,!,
`Zo
`M: E
`~:::l
`0 I-
`
`5
`
`0 -
`
`/...... 1-....f:
`:::,...
`t--.
`~o Y & &~
`§ Q_Q I:::;:::~
`u ~ ~ ~~
`<:-
`i:::-
`i:::-
`zzz ~
`t--.
`y
`tP
`·s
`~ z
`
`60
`
`40
`
`ti)
`...I
`j:::
`+
`'q'
`C
`u
`
`20
`~
`0
`
`n.d.
`
`0
`
`t--.
`
`c5"
`/......
`/......
`:.$ y & &
`§ tP I:::
`I:::
`·s.
`U
`•s
`~s
`f' z z
`:--.:; ~ ~
`
`~
`t--.
`y
`Q_(j
`
`~ f'
`
`Merck Ex. 1005
`Page 21 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 21 of 65
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`US 2015/0216970 Al
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`FIG. 110-11 F
`
`D
`
`E
`
`F
`
`C
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 22 of 65
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`US 2015/0216970 Al
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`FIG. 12A-12C
`
`n
`
`A
`VI 40-(cid:173)
`..J
`j::
`+ 30
`00
`0
`U 20
`+
`i'tl
`~ 10
`I-
`
`n
`
`B
`
`30·--
`
`(/)
`..J
`j:: 20-
`+
`00
`0
`U 10
`?ft.
`
`C
`j 100-·(cid:173)
`j::
`+ 80
`00
`0
`U 60
`"O
`,! 40
`
`i'tl > t; 20
`<
`?ft.
`
`*
`
`0 _..._..,.............,. ....... ..,..........._ .....
`
`Merck Ex. 1005
`Page 23 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 23 of 65
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`US 2015/0216970 Al
`
`FIG. 13A-13E
`
`A
`
`Control
`
`B
`
`Anti-PDL1
`
`C
`
`Anti-TIGIT
`
`6%
`
`ftb:"mfflffl""., ~
`CDB
`
`12%1--:c~il
`tf}
`
`E
`
`o:=i!::::!:::: E
`co __ (/)
`~ I (.90:..::;
`00..1--+- o
`0 . ..!..:6:6z
`.._,cc
`~<(<(
`+
`
`Merck Ex. 1005
`Page 24 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 24 of 65
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`US 2015/0216970 Al
`
`FIG. 13F-13I
`
`F
`
`G
`
`H cos+ TILs
`45,,
`
`+,__ 30 ·
`z
`!a!:
`'?f. 15
`
`CD4+T!Ls
`
`+,__ 30·
`z
`!!,:
`'?f. 15
`
`No
`Stirn.
`
`Anti-POL 1
`Anti-TIGIT -
`
`-
`+
`+
`- + + -
`
`No
`Stirn.
`
`+
`...
`- + 1
`
`-
`
`0
`Anti-POL 1
`Anti-TIGIT -
`
`Merck Ex. 1005
`Page 25 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 25 of 65
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`US 2015/0216970 Al
`
`• ••
`
`,
`
`I
`
`I 1,
`
`Splenic B Cells
`------ Splenic CD8+ T Cells
`------- CD8+ TIGIT + Tl Ls
`
`.~ - ,1
`:~ ! ~·\p
`~ I
`i ~
`i ?
`I I
`!
`~l:i
`i !!
`i
`j :!
`:
`i !?
`:
`,· :,,
`:
`i :~:
`:
`J'.
`; \t:
`: \,",:
`; i:
`... , "
`,,
`
`I
`
`1•,1
`
`'
`I
`'
`'
`I
`
`,,
`I'
`(cid:127):
`\ :
`
`CD226
`
`FIG.14
`
`pRK
`
`Tigit+CD226
`
`IP Anti-HA
`
`IP Anti-Flag
`
`+ + - -
`
`- - + -
`
`+ - + -
`
`- + - -
`
`+ + - -
`
`- - + +
`
`+-+-
`
`- + - +
`
`._TIGIT
`
`Anti-Flag H RP
`(CD226)
`
`Anti-HA-H RP
`(TIGIT)
`
`FIG.15
`
`Merck Ex. 1005
`Page 26 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 26 of 65
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`US 2015/0216970 Al
`
`1
`
`2
`
`3
`
`4
`
`._CD226
`
`FIG. 16
`
`Merck Ex. 1005
`Page 27 of 140
`
`

`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 27 of 65
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`US 2015/0216970 Al
`
`FIG. 17A-17F
`
`A
`
`0
`-..:;
`(V
`rx:
`I(cid:173)
`LU
`rx:
`LL
`
`D
`
`FRET ST-CD226 I ST-CD226
`1.5 ..
`
`B
`
`0
`3
`10
`50 100
`5 ng Flag-ST-CD226
`+ HA-TIGIT (ng)
`
`C
`
`FRET ST..CD226 / ST-CD226
`._ 6 0 .6 - - - - -
`ns
`>, 'iij
`:!: Ill
`1// Q)
`C: a,0.4··
`..!! ><
`..:: LU
`t
`1- I- 0.2
`wt?
`rx: C")
`~
`LL ~ 0 0 --.1..J....~L....J..-.U
`LL
`.
`~~
`Anti-T!GIT PBS
`Flag-ST-CD226
`HA-TIGIT
`
`FRET HA-TIGIT I ST-CD226
`
`E
`
`F
`
`FRET ST ..CD226 / HA-TIGIT
`**
`
`C:
`
`-- 0 ~·w
`'iii~ 2
`~a
`.., X
`-= w
`I- 1-woo
`a:::
`'
`LI.. g>
`u: o-u.....,...1.....1._.1.1
`Anti-T!GIT PBS
`
`1 ··
`
`Merck Ex. 1005
`Page 28 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 28 of 65
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`US 2015/0216970 Al
`
`4x1o06
`
`FIG.18
`
`EUSA EJ Flag
`D HA
`
`0
`
`3
`
`100
`
`5 ng Flag-ST..CD226
`+ HA-TIGIT (ng)
`
`-3x1QOB
`
`.,,2x10°6
`
`0
`
`:c
`>
`J;'
`3
`:I
`i
`£ ::,
`·· 1 x10°6 .!
`?J
`C
`.:...,.
`
`Merck Ex. 1005
`Page 29 of 140
`
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 29 of 65
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`US 2015/0216970 Al
`
`FIG. 19A-19D
`
`A
`
`B
`
`-c,$
`
`0 -.... . a> ~u
`:.::;f-
`(J +
`<(m
`:zeO
`¢ 0
`
`80
`
`60
`
`40
`
`**~
`
`Anti-POL 1 -
`Anti~TIGIT -
`Anti-CD226 -
`
`+ +
`+ +
`+
`-
`
`+
`
`-
`Anti-POL 1 -
`+ +
`• + +
`Anti-TIGIT -
`Anti-CD226 - +
`- +
`
`D
`
`C
`
`25
`
`..... .!!!
`o-:n;
`+o 15·
`~I-
`I.I..+ H}
`-ro ~o
`"'u
`
`c,......,........,......,..,........_
`+ +
`-
`-
`Anti-PDL1
`+ +
`-
`-
`Anti-TIGJT
`+
`-
`+
`Anti-CD226 -
`
`-
`Anti-POL i
`-
`Anti-TIGIT
`Antl-CD226 -
`
`-
`-
`+
`
`+ +
`+ +
`+
`
`Merck Ex. 1005
`Page 30 of 140
`
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 30 of 65
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`US 2015/0216970 Al
`
`FIG. 20A-20D
`
`B
`
`D
`
`lUSC
`
`p~ 0.82
`
`2
`
`10 50
`TIGIT
`
`500
`
`COAD
`
`A
`
`5000-
`
`1000-
`
`<,I,)
`~
`C
`0
`
`200···
`
`50-·
`
`C
`
`1000
`
`200·
`
`I.A)
`C")
`Cl
`u
`
`10
`
`0
`
`0
`
`p= 0.83
`
`5
`
`20
`TIGIT
`
`200
`
`•
`1 . v
`.,
`
`0
`~
`
`(I,)
`M
`0.2···
`0
`0
`..... 0.1·-
`C)
`i=
`
`0.05-·
`
`•
`•
`
`t2. 0.5-· I
`!
`-
`
`***
`
`&·
`~
`'\0~ ~o~
`
`0.5
`
`0
`:-.;:::;
`ro
`n:::
`0.2·--
`~
`Cl
`u
`I-
`a
`j::: 0.05
`
`0.1
`
`***
`
`i
`
`•
`•.
`•
`~
`o\
`A,._0~ ~&~
`
`Merck Ex. 1005
`Page 31 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 31 of 65
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`US 2015/0216970 Al
`
`FIG. 20E-20H
`
`E
`
`2000-
`
`500
`
`100
`
`20
`
`5
`
`"'
`M
`C
`u
`
`UCEC
`
`F
`
`2
`
`0.5
`
`0.2
`
`0
`;:;
`t}_
`ti)
`M
`C
`u
`I-
`c3
`.:;: 0.05
`
`0.1
`
`0
`
`p= 0.87
`
`0.02
`
`1 2
`
`50
`10
`TIGIT
`
`500
`
`G
`
`BRCA
`
`~
`C
`u
`
`10
`
`1 ...
`
`(l
`
`p= 0.86
`
`510 50 500
`TIGIT
`
`H
`
`5
`
`0
`:.::;
`(ti
`0:::
`ti) 0.2 ..
`M
`C
`u
`-
`I-
`c3
`.:;: 0.01 ..
`
`005··
`.
`
`•
`
`•
`
`***
`
`~
`
`-
`='
`& ~
`,<.._v<$' ~&~
`..
`
`•
`•
`
`1rl<*
`
`~
`
`I
`'
`
`•
`& ~
`;<,.._v~ ~a~
`
`Merck Ex. 1005
`Page 32 of 140
`
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 32 of 65
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`US 2015/0216970 Al
`
`0.5
`
`0.1 ···
`
`0
`z
`nJ
`0:::
`fl>
`M
`C
`U 0.05 ..
`!:: 0.02
`C)
`j:: 0.01···
`
`FIG. 201-20L
`
`KIRC
`
`p= 0.94
`
`510 50
`TIGIT
`
`500
`
`J
`
`L
`
`t.;i
`(")
`C
`u
`
`5000
`
`1000
`
`200
`50 ·
`
`10
`
`K
`
`10000·
`< 2000
`00
`C
`500
`0
`
`100
`
`0
`
`0.2 ... I t
`
`•
`,.
`•
`
`• ..
`
`•
`•
`
`o'
`~
`~~
`~o
`
`.4....v<f::'
`
`10000
`
`2000
`
`500
`
`100
`
`-.::I"
`C
`0
`
`I
`
`2 10 50 2001000
`TIGIT
`
`2 10 50 200 1000
`TIGIT
`
`Merck Ex. 1005
`Page 33 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 33 of 65
`
`US 2015/0216970 Al
`
`FIG. 20M-20N
`
`0 Tumor
`•Normal
`
`o
`
`0
`
`M
`
`100---o 20--
`
`a..
`
`N
`
`500·
`
`100 --
`
`20·
`
`5---
`
`(0
`N
`N
`Cl
`0
`
`0
`
`0
`
`0p= 0.82
`
`I
`I
`10 50
`TIGIT
`
`500
`
`1-- 0
`I
`2
`
`0
`
`p= 0.64
`
`I
`10 50
`TIGIT
`
`I
`
`500
`
`Merck Ex. 1005
`Page 34 of 140
`
`

`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 34 of 65
`
`US 2015/0216970 Al
`
`CO2
`SLA2
`SIRPG
`CXCR6
`CD24l
`P2RY10
`ITK
`CXCR3
`GPR171 UBASH3A
`LOC100506776
`SH2D1A GPR174
`cog6
`SLAMF6
`TIGIT
`IL 12RB 1
`LCK TBC1D10C
`NKG7 LINC00426
`CST7
`PTPRC
`PTPN22 GIMA P5
`GIMAP1-GIMAP5sfff
`PYHIN1 ARHGAP15
`SASH3--
`GIMAP4
`GIMAP7
`CRTAM
`SLAMF1
`PDCD1
`GVINP1
`CD6
`IL2RG
`CCR5
`LOC100652927 TESPA 1
`IL10RA
`GZMH
`PCED1B-AS1
`CTLA4
`SCML4
`CCL5
`CD48--
`PTPRCAP ~
`
`SIGNATURE ---[[[fllillw,jM ..... : 1 ...... 1 ..... m ... 1tt1_1 ----11,___,;,...___,;,... _______ .;.;;.1 .... 1 -(cid:127)
`
`-·-111111.11 .. w-_
`
`!I
`
`j J
`!
`l j
`'
`i' ~ '! li
`
`~
`I
`
`.
`
`i
`
`Ii
`ij!' H.
`•
`
`. "
`
`I
`fj
`
`:1 r'.1
`
`I
`
`II
`I
`
`!!i
`I l
`!If I
`
`iJ
`
`i
`
`FIG. 21
`
`Merck Ex. 1005
`Page 35 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 35 of 65
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`US 2015/0216970 Al
`
`FIG. 22A-22F
`
`A
`
`PBMC (Norma!)
`
`B PBMC (NSCLC)
`
`C
`
`Tll {NSCLC)
`
`10%
`
`42%
`
`51%
`
`I- -
`gU:.,...,., .,...............,....
`CD8
`
`D
`
`PBMC {Norma!)
`
`E
`
`PBMC (NSCLC)
`
`F
`
`TIL {NSClC)
`
`17%
`
`17%
`
`39%
`
`!:::
`
`~ f:: ...... , ,.,...-......-...,...,...
`
`CD4
`
`Merck Ex. 1005
`Page 36 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 36 of 65
`
`US 2015/0216970 Al
`
`..c
`CD 5 0)
`..c
`0. 0
`c~~
`000
`(/) Q_ Q_
`
`+
`~
`0
`()
`
`~ .c
`0 0)
`Q_ :.c
`.-·
`I
`____ ./ _,,
`__ .. ____ .....z::-~•= ..... =~-::::_ - - -
`----·----- .. - .. _ ...
`..... --
`......
`.. _____ ~
`.,,,.,.
`~
`
`-
`-
`_-::.,..,
`
`I..
`
`1-(cid:173)
`(9
`I-
`
`.c
`0)
`.c
`
`+ co
`0 u
`
`~
`I
`0
`.---
`-------
`Q_
`,,. -
`--------•-----·-
`.. ________
`,,,
`,. - --_:-:. .. -=-~.::_:-_~_
`
`Merck Ex. 1005
`Page 37 of 140
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 37 of 65
`
`US 2015/0216970 Al
`
`FIG. 221-22L
`
`*
`
`75
`
`J +
`co
`C
`U !!!. 50
`+-1::B
`
`(!) I- 25
`I-
`
`K
`
`L
`
`26%
`
`17
`LnJ
`
`; '. /~\,/ , ~:<
`
`75
`
`+ ,q'
`C
`U!!!. 50···
`+-
`!::~
`(!) I- 25
`j::
`
`***
`
`1-
`
`~ C''
`
`CD4
`
`Merck Ex. 1005
`Page 38 of 140
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 38 of 65
`
`US 2015/0216970 Al
`
`FIG. 22M-22P
`
`M
`
`Spleen
`---- TiLPD-1k)W
`----.----T!L PD-1high
`cos+
`
`TIGIT
`
`0
`
`Spleen
`-- -- TiL P0-1low
`------ T!l PD-1high
`co4+
`PD-1high
`
`TIGff
`
`N
`
`900···
`
`u:: 600···
`::!:
`I::
`C, r= 300···
`
`p
`
`1200
`
`U:: 800···
`::IE
`t-
`(5
`j:: 400 ...
`
`Merck Ex. 1005
`Page 39 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 39 of 65
`
`US 2015/0216970 Al
`
`FIG. 23A-23D
`
`A
`
`Tlls (NSCLC)
`
`s
`
`76%
`
`76%
`
`1-
`
`~ U:.,...,..,.........,.................,..............., ~ D ...... · ,........,....,..........,....,.............,.....
`
`1-
`
`CD8
`c PBMC (NSCLC)
`
`CD4
`
`D
`
`47%
`
`40%
`
`I-
`
`~ t_t~--,--,--
`CD8
`
`I-
`~ tt.,.,.., ......-..-,---.-.-~,-I
`CD4
`
`Merck Ex. 1005
`Page 40 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 40 of 65
`
`US 2015/0216970 Al
`
`FIG. 23E-23H
`
`E
`
`TILs (CRC)
`
`F
`
`53%
`
`47%
`
`r-
`r-
`~ U:----- ~ U:-----
`
`CDS
`
`CD4
`
`G
`
`PBMC (CRC)
`
`H
`
`27%
`
`r-
`-
`~U:---
`
`CD4
`
`Merck Ex. 1005
`Page 41 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 41 of 65
`
`US 2015/0216970 Al
`
`PBMC (CRC)
`
`27%
`
`FIG. 23D
`
`TIGIT:CD226
`25--------------
`FRET ratio
`
`20
`
`15
`
`10
`
`5
`
`1 00
`50
`1 0
`1 00
`50
`1 0
`0
`Human HA-TIGIT (ng) Human HA-TIGIT Q56R (ng)
`
`FIG. 24
`
`Merck Ex. 1005
`Page 42 of 140
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`Aug. 6, 2015 Sheet 42 of 65
`
`US 2015/0216970 Al
`
`FIG. 25A
`
`o Control
`A aTIGIT
`(cid:143) o:PD-L1
`v o:TI GIT + czPD-L1
`
`-~--R
`
`1 00 .,
`~V,.__
`~~z,,,_51.
`;
`: - - - -~ - - - .. , - - -- ~- - -~- - -- - - - - \~J;;/~~- -
`Complete Response (CR)
`10 -----.., - - - - - - - - - - - - - - -
`20
`40
`60
`0
`
`Day
`
`Merck Ex. 1005
`Page 43 of 140
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 43 of 65
`
`US 2015/0216970 Al
`
`...... ~
`
`~!''~' _-.-._-,-~t:!1-.o'I'!'!""~--(-· 0
`
`0 ---
`
`0
`N
`
`fflll"""'• ~i--11"""""'.::ll~"'-"""""""..,.1 .... 0
`0
`0
`8
`
`0 - 0
`
`""""'
`
`()
`
`w
`
`····i
`
`g
`c::
`0 u
`
`""r ""'' """'·· """"""""""'"""""'!l(;J;I""',-""'·· ""·' ~-t1· ··· O
`0
`
`~
`
`C
`
`w
`LO
`CN ca
`
`LO
`CN
`ci
`u:::
`
`[D
`
`Merck Ex. 1005
`Page 44 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 44 of 65
`
`US 2015/0216970 Al
`
`FIG. 25F-25G
`
`F
`
`G
`
`;;--- 10000
`E
`!.
`E 1000
`::,
`j
`i..
`0
`E
`t=
`
`(I.I
`
`'¢
`T"
`
`100
`
`~
`Q
`
`10
`Anti-POL 1
`Anti-Tl GIT
`
`+
`
`+
`+
`
`+
`
`100-----~-~-~
`
`c1·
`
`ro 1s-
`
`::i
`
`0-25
`
`' 1 o Contr_o_ !
`-~
`~1·}-l-------v
`c i7
`i SO
`6 ~ ::~+oPDL1
`h 7
`0 ____ :J_l ______ __,
`
`A aTlGIT
`
`0
`
`20
`
`40
`
`60
`
`Day
`
`Merck Ex. 1005
`Page 45 of 140
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 45 of 65
`
`US 2015/0216970 Al
`
`FIG. 26A-26H
`
`A
`
`B
`
`Resting
`
`C -------
`
`Hours
`
`TIGIT
`
`E
`
`t:
`~o
`
`CDS
`
`67%)
`
`[j
`
`\
`:?!t
`
`TIGIT
`
`G
`
`***
`
`TIGIT
`
`F
`
`80
`
`.... 1/)
`o=
`Cl) 60-
`+
`i,-U
`-i,-
`~+
`... co
`~o
`C 0
`
`40
`
`20-
`
`0 i J.
`
`0~0 -<...-,)<$'"
`
`D
`
`300
`250
`U: 200-
`:!E
`i,- 150
`a 100
`.:::
`
`H
`
`60
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 46 of 65
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`US 2015/0216970 Al
`
`FIG. 261-26L
`
`Spleen
`- -- - TIL PD-1 1(cid:144)W
`---- TIL PD-1high
`
`J
`
`900-
`
`i 60fJ--
`t-
`§ 300
`I-
`
`Spleen
`--- - TIL P0-1low
`- - TIL PD~1high
`
`K
`
`L 000
`
`TIGIT
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 47 of 65
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`US 2015/0216970 Al
`
`FIG. 27A
`
`Spleen
`
`Tumor
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 48 of 65
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`US 2015/0216970 Al
`
`u
`
`I
`
`,
`
`Merck Ex. 1005
`Page 49 of 140
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`

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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 49 of 65
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`US 2015/0216970 Al
`
`FIG. 28A-28F
`
`A
`
`-<>- uCD226 + aPD-L 1 + u TIGIT
`-o- Control
`-n-aCD226
`-o-aPD-L 1
`---s:J-- a TIGIT + aPD-L 1
`ME
`..S 10000------------,
`(I)
`E
`:::l
`~
`0
`E
`i2
`s:::
`~ 10-,
`0
`(I)
`2
`
`8
`c
`Control
`aPD-L1
`10000-----------.10000,._ - - - - - - - - - ,
`
`10 1
`0
`
`10
`
`20
`
`30
`
`40
`
`50
`
`1 0 - - - - - - - - - - '
`0
`10
`20
`30
`40
`50
`
`100
`
`-
`
`-
`
`10
`
`-- ~-
`
`30
`
`40
`
`20
`Day
`
`D
`a.CD226
`10000-, - - - - - - - - - - ,
`
`E
`
`a TIGIT +aPD-L 1
`
`F
`
`ctPD-L 1 +a TIGIT +aCD226
`
`10------~,---1
`0
`10
`20
`30
`40
`50
`
`10
`
`20
`
`30
`
`40
`
`50
`
`10-l-,---------!
`0
`10
`20
`30
`40
`50
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 50 of 65
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`
`FIG. 28G-28K
`
`G
`
`-o-o:PD-L1+oTIGIT +aCD226
`--o- Control
`-t.-aCD226
`-o-uPD-L1
`-:zJ- oJIGIT +o;PD-L1
`
`H
`
`100·
`
`t5
`~
`i::
`::I
`
`ti) -C:
`(l) u ... (l) a..
`
`50
`
`0 ---...----.'.l'-::,.,---1:::J!
`20
`10
`0
`30
`
`Anti~POL 1 - + - + + -
`Anti-TIG!T
`- + +
`-
`Anti-CD226 -
`- + - + -
`
`J
`3{1--------
`**
`~
`i= 20
`
`••
`
`+ «:, S lO··
`o..._ ...... ......., .......................
`Antl-POL 1 - +
`+ +
`-
`- + +
`Anli-TIGIT -
`-
`Anti-CD226 -
`- + -
`+
`
`Anti-POL 1 -
`Anti-TIGIT -
`Anti-CD226 -
`
`+ +
`-
`+
`- + +
`-
`- + - +
`
`Anti-POL 1 - + - + + -
`Anti-TIGIT -
`+ + -
`Anti-CD226 -
`-
`t
`-
`
`- +
`
`Merck Ex. 1005
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 51 of 65
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`US 2015/0216970 Al
`
`FIG. 29A-29D
`
`A
`
`Murine CD84 T Cells
`
`B
`
`M urine Cos+ T Cells
`
`Hr,
`
`g,i-i,,
`
`- +
`
`2i
`50
`~ o 40
`~la 30 = 0::: 20-
`e
`10
`a..
`IJ--i..,-.~~~:,;.u::.,..i...t.,..u
`Anti-CD3 - -+
`-+ + + + +
`mPVR-Fc -
`+ + t
`t
`Anfi*TlG!T -
`- + -
`+
`Anti-CD226 -
`- - + +
`Human C0226
`
`~---~
`~:,~
`
`C
`
`D
`
`Human coa+ T Cells
`
`,.,,......., .•. """ /~~~--,
`
`l:(cid:173)
`·~ <if 40--
`.9? g. 30
`..EC")w' 20•••
`!--0
`W!:., 10
`ff
`0-+-----.i--------
`rn
`oo
`0
`20
`Donor Emission
`(103,620 nm)
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 52 of 65
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`US 2015/0216970 Al
`
`FIG. 29E-29F
`
`E
`
`F
`
`--------... 88 t HA 8
`
`0.4 -~------,
`
`c.o 0 "'·(cid:173)
`c--.a-
`ro
`C
`ua::
`-1-
`!::::w
`(!) a::
`i= LL.
`
`6---------
`4
`
`2
`Q _..._.,..i ~.,.._""""-,-1........,.~i u
`0
`10
`50
`100
`Human TIGIT (ng)
`
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 53 of 65
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`US 2015/0216970 Al
`
`FIG. 29G-29H
`
`G
`
`H
`
`TIGIT + CD226
`+ +
`TIGIT IP - + + -
`CD226 IP + -
`- +
`CD226 IB
`
`+ +
`- + + -
`+ -
`- +
`TIGITIB
`
`-
`
`~ 60 kDa
`
`~ 26 kDa
`
`*** nl
`
`Q--L...i..,..&.-.i:,;;.,:a....11
`
`Anti-TIGIT
`
`+
`
`-
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 54 of 65
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`US 2015/0216970 Al
`
`;?::'
`u, -
`s::::: u,
`
`C1) a. - (.)
`w""""'" ~(cid:173)
`
`s:::::
`-
`-M
`I- 0
`
`LL
`
`2000
`1500
`1000
`500
`0----1-"-.................................... "'----,,,,...,..,..,,..,,T,t
`-500---------- ___, _____ _
`Acceptor Control +
`Donor aJIGIT +
`Unlabeled a TIGIT
`-
`Acceptor aCD226
`-
`
`CD226+ TIGIT +
`***
`***
`
`CD226+ TIGIT-
`
`--lf--L-'-'--......._............,;......._ __ ___
`
`+
`+
`
`+
`
`+
`
`+
`+
`+
`
`+
`
`+
`
`+
`+
`+
`
`FIG. 30
`
`Merck Ex. 1005
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 55 of 65
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`US 2015/0216970 Al
`
`FIG. 31 A-31 B
`
`A
`
`B
`
`00
`
`u, -
`(I.) 9
`u
`I-
`+ 6
`C u
`-
`....
`«s 3
`~
`~ 0
`- + - +
`Anti-POL 1
`- + +
`Anti-TIGIT -
`
`0
`
`0
`
`3
`
`**
`
`o en
`t.==
`· - (I.)
`~u
`C.1- 2
`cc>+
`Meo
`Mc 1
`c..u
`(!) '4-
`~o
`0
`Anti-POL 1
`- + - +
`- + +
`Anti-TIGIT -
`
`Merck Ex. 1005
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 56 of 65
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`US 2015/0216970 Al
`
`FIG. 31 C-31 G
`
`C
`
`Control
`
`E
`
`D
`
`Anti-PD-L1
`ylsci
`24%
`
`F Anti-PD-L 1
`+ Anti-TIGIT
`
`!tJffii]
`
`59%
`
`**
`
`..!!!
`,;, cii
`G MU 60--
`0. t(cid:173)
`C> +
`ti~ 40
`+(.)
`;,-0
`:z: ;;:: 20
`~-y
`~~
`00 0~!4J..t~~
`Anti-POL 1 - + - +
`Anti-TIGIT -
`- + +
`
`....
`
`19%
`
`~lb:'· c, ....,.,...,...,..,,........,,,.....,.
`
`a>
`a.. IFNy
`
`Merck Ex. 1005
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 57 of 65
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`US 2015/0216970 Al
`
`FIG. 32A-32D
`
`A
`
`-o- Control
`-0- Treated, CDS-depleted
`-f'r Treated, No depletion
`
`Complete Response (CR)
`
`10
`
`0
`
`10
`
`20
`
`Day
`
`30
`
`B
`
`10000
`
`100
`
`10
`
`------------------
`
`0
`
`10
`
`20
`
`30
`
`C
`
`10000
`
`1000
`
`100
`
`10
`
`0
`
`Treated, no Depletion
`
`10
`
`20
`
`30
`
`D
`
`10000
`
`1000
`
`100
`
`10
`
`0
`
`10
`
`20
`
`30
`
`Merck Ex. 1005
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 58 of 65
`
`US 2015/0216970 Al
`
`FIG. 32E-32F
`
`E
`
`F
`
`10000,
`
`1000''.
`
`100-
`
`***
`
`i ~
`
`Do
`
`,,.
`
`M
`E
`.._ E
`o_
`E Q)
`~E
`:::I
`~
`
`10-
`anti-TIGIT/PDL 1 •· + +
`- +
`CDS depletion -
`
`I
`
`I
`
`10000 Naive Control
`
`1000
`
`100 C'
`
`101-1 -
`0
`
`....... - - - - , - - - ,
`15
`10
`20
`5
`
`Merck Ex. 1005
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 59 of 65
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`US 2015/0216970 Al
`
`FIG. 32G-321
`
`H
`10000
`
`CR, CDS-depleted
`
`G
`
`1000
`
`-o- Naive
`v CR, CDS-depleted
`-tr CR, No Depletion
`
`-----~ 10-----....,......-.....,.....---,
`~
`
`1000
`
`100
`
`0
`10000-,,
`
`5
`
`10
`
`15
`
`20
`
`CR, No Depletion
`
`- - - - - - - - -~
`Complete Response (CR)
`10_., ___________ __ _
`
`0
`
`5
`
`10
`Day
`
`15
`
`20
`
`100 ~
`' - - - - -~
`10-----....,......-.....,.....---,
`10
`15
`20
`5
`0
`
`Merck Ex. 1005
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 60 of 65
`
`US 2015/0216970 Al
`
`o Wildtype, Untreated
`
`(cid:143) Wildtype, aTIGIT/PD-L 1
`v PVR.KO, aTIGIT/PD-L 1
`
`10000
`
`s...-OM 5 ~ 1000
`
`1-(cid:173)
`s:::: CL)
`.!!! E
`-g ~ 100
`:E:~
`
`10---+--------,-------------,
`10
`20
`30
`0
`
`Day
`
`10000 Wildtype, Untreated
`
`10000 PVR.KO, Untreated
`
`1000
`
`100
`
`1000
`
`100
`
`10---+---.....---------,
`10
`20
`30
`0
`
`10---+---.....---------,
`10
`20
`30
`0
`
`10000 Wildtype, aTIGIT/PD-L 1 10000 PVR.KO, aTIGIT/PD-L 1
`
`10---+---.....---------,
`10
`20
`30
`0
`
`10---+---.....--------,
`10
`20
`30
`0
`
`FIG. 33
`
`Merck Ex. 1005
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 61 of 65
`
`US 2015/0216970 Al
`
`-0- Control
`-a- aTIGIT
`-t:r aPD-L 1
`..s;;.- a TIGIT + aPD-L 1
`
`10000
`
`s...-OM
`~ ~ 1000
`I- -
`C: Cl)
`-~ E
`~..:! 100
`:E~
`
`Complete Response (CR)
`10_...,_ ____ .....,. _________ ___,
`
`0
`
`10
`
`20
`
`30
`
`40
`
`50
`
`Day
`
`10000
`
`1000
`
`10000
`
`1000
`
`100
`10_...,. _______ ..,......____,
`
`100
`10_...,. _______ ..,......____,
`
`0
`
`1 0
`
`20
`
`30
`
`40
`
`50
`
`0
`
`1 0
`
`20
`
`30
`
`40
`
`50
`
`10000
`
`1000
`
`10000
`
`1000
`
`100
`10_...,. _______ ..,......____,
`
`100
`10_...,. _______ ..,......____,
`
`0
`
`1 0
`
`20
`
`30
`
`40
`
`50
`
`0
`
`1 0
`
`20
`
`30
`
`40
`
`50
`
`FIG. 34
`
`Merck Ex. 1005
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`

`Patent Application Publication
`
`Aug. 6, 2015 Sheet 62 of 65
`
`US 2015/0216970 Al
`
`B
`
`+
`~
`u..
`z
`I-
`+
`~-
`z
`u..
`cf!.
`
`dlN CD4+ T Celis
`
`15,,
`
`10--
`
`5-
`
`No
`Stirn.
`
`FIG. 35A-358
`
`A
`
`dlN cos+ T Cells
`
`**
`
`15-
`
`+
`;:$
`u.. 10--
`z
`I-
`+
`>-
`z
`u..
`'#-
`
`5-
`
`0- /:
`Anti~PDL 1 - + - + -
`- + + -
`Anti-T!GIT -
`
`[}---
`
`I
`
`Antl-PDL 1 - + - +
`Anti-TIGIT -
`- + + -
`
`~
`
`Merck Ex. 1005
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`Patent Application Publication
`
`Aug. 6, 2015 Sheet 63 of 65
`
`US 2015/0216970 Al
`
`FIG. 35C-35D
`
`C
`
`cos+T1Ls
`
`D
`
`20
`
`co4+ TILs
`
`+ Ci 15-··
`lL z
`1-
`..J..
`J0'-z u,,.
`
`0~
`
`5····
`
`10·
`
`No
`Stirn.
`
`o_y;;.....,_....,.i...~~~ .....
`
`Anti-POL 1 - + - + -
`Antl-TIGIT -
`- + + -
`
`O·------
`
`Anti-PDL 1 - + - + -
`- + + -
`Anti-TIGIT -
`
`Merck Ex. 1005
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`
`Aug. 6, 2015 Sheet 64 of 65
`
`US 2015/0216970 Al
`
`FIG. 36A-36B
`
`A
`
`B
`
`*
`
`t,.
`
`* f
`~ t,.
`(cid:143)
`
`!!!. 100···
`Q)
`(.)
`I-
`+
`00
`C
`(.)
`
`80·
`
`60···
`
`-
`
`40·-
`
`0
`
`0
`+
`!::
`20- %
`C)
`j::
`~ 0
`
`!!!.
`Q)
`(.)
`I-
`+
`-.::I"
`C
`(.)
`
`-
`
`0
`
`iOO ···
`
`80
`
`60···
`
`20··
`
`t
`40- ~
`
`t,.
`
`t,.
`
`-(JD
`
`(cid:143)
`
`I
`
`0
`+
`!::
`C)
`j::
`~ 0
`
`~ ~0 -<-.o/
`~&-<S- <l<>
`
`~ ~0 -<-.o/
`~&<S' q_<;j
`
`Merck Ex. 1005
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`Patent Application Publication
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`Aug. 6, 2015 Sheet 65 of 65
`
`US 2015/0216970 Al
`
`..!!?
`(1)
`(.)
`I
`Lt)
`"1::1'
`C
`(.)
`
`..!!?
`(1)
`(.)
`'C
`:§
`(1)
`>,
`:i::
`+ Lt)
`--:t'
`C
`(.)
`
`f/J
`(I)
`
`.... >,
`
`0
`0
`.c:
`0.
`E
`~
`+
`Lt)
`--:t'
`C
`(.)
`
`u
`
`cc
`
`<(
`
`w
`r-,..
`M
`<i:
`r-,..
`M
`ci
`u::::
`
`1-
`I
`I
`
`!::
`t!)
`i==
`
`!::
`t!)
`j::
`
`!::
`t!)
`j::
`
`...J
`i=
`+
`"'11:2"
`0
`0
`0
`...J
`0
`U)
`:z
`
`w
`
`...J
`I-
`+
`00
`0 u
`u
`...J
`0
`U)
`:z
`
`C
`
`:l]s
`
`~-Od
`
`djs
`
`~-Od
`
`Merck Ex. 1005
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`US 2015/0216970 Al
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`Aug. 6, 2015
`
`METHODS OF TREATING CANCER USING
`PD-1 AXIS BINDING ANTAGONISTS AND
`TIGIT INHIBITORS
`
`CROSS REFERENCES TO RELATED
`APPLICATIONS
`
`[0001] This application claims the priority benefit of U.S.
`Provisional Application No. 61/846,941, filed Jul. 16, 2013,
`U.S. Provisional Application No. 61/865,582, filed Aug. 13,
`2013, U.S. Provisional Application No. 61/950,754, filed
`Mar. 10, 2014, U.S. Provisional Application No. 61/985,884,
`filed Apr. 29, 2014, and U.S. Provisional Application No.
`61/992,109, filed May 12, 2014, each of which is hereby
`incorporated by reference in its entirety.
`
`SUBMISSION OF SEQUENCE LISTING ON
`ASCII TEXT FILE
`
`[0002] The content of the following submission on ASCII
`text file is incorporated herein by reference in its entirety: a
`computer readable form (CRF) of the Sequence Listing (file
`name: 146392025900SEQLISTING.TXT, date recorded:
`Jul. 16, 2014, size: 25 KB).
`
`BACKGROUND OF THE INVENTION
`
`[0003] The provision of two distinct signals to T-cells is a
`widely accepted model for lymphocyte activation of resting T
`lymphocytes by antigen-presenting cells (APCs). Lafferty et
`al,Aust. J. Exp. Biol. Med. ScL 53: 27-42 (1975). This model
`further provides for the discrimination of self from non-self
`and immune tolerance. Bretscher et al, Science 169: 1042-
`1049 (1970); Bretscher, P.A., P.N.A.S. USA 96: 185-190
`(1999); Jenkins et al, J. Exp. Med. 165: 302-319 (1987). The
`primary signal, or antigen specific signal, is transduced
`through the T-cell receptor (TCR) following recognition of
`foreign antigen peptide presented in the context of the major
`histocompatibility-complex (MHC). The second or co-stimu(cid:173)
`latory signal is delivered to T-cells by co-stimulatory mol(cid:173)
`ecules expressed on antigen-presenting cells (APCs), and
`induce T-cells to promote clonal expansion, cytokine secre(cid:173)
`tion and effector function. Lenschow et al., Ann. Rev. Immu(cid:173)
`nol. 14:233 (1996). In the absence of co-stimulation, T-cells
`can become refractory to antigen stimulation, which results in
`a tolerogenic response to either foreign or endogenous anti(cid:173)
`gens.
`[0004]
`In the two-signal model, T-cells receive both posi(cid:173)
`tive co-stimulatory and negative co-inhibitory signals. The
`regulation of such positive and negative signals is critical to
`maximize the host's protective immune responses, while
`maintaining immune tolerance and preventing autoimmunity.
`Negative signals seem necessary for induction of T-cell tol(cid:173)
`erance, while positive signals promote T-cell activation.
`[0005] Both co-stimulatory and co-inhibitory signals are
`provided to antigen-exposed T cells, and the interplay
`between co-stimula