(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2009/0258013 A1
`Clark et al.
`(43) Pub. Date:
`Oct. 15, 2009
`
`US 20090258O13A1
`
`(54) NOVEL COMPOSITIONS AND METHODS
`FOR THE TREATMENT OF IMMUNE
`RELATED DISEASES
`
`(75) Inventors:
`
`Hilary Clark, San Francisco, CA
`(US); Dan Eaton, San Rafael, CA
`(US); Lino Gonzalez, JR., Menlo
`Park, CA (US), Jane Grogan, San
`Francisco, CA (US); Jason A.
`Hackney, Palo Alto, CA (US);
`Kristin Harden, Pacifica, CA (US);
`
`Correspondence Address:
`GENENTECH, INC.
`1 DNAWAY
`SOUTH SAN FRANCISCO, CA 94080 (US)
`
`(73) Assignee:
`
`Genentech, Inc., South San
`Francisco, CA (US)
`
`(21) Appl. No.:
`
`12/420,234
`
`(22) Filed:
`
`Apr. 8, 2009
`Related U.S. Application Data
`(60) Provisional application No. 61/194.271, filed on Sep.
`26, 2008, provisional application No. 61/123.530,
`filed on Apr. 9, 2008.
`Publication Classification
`
`(51) Int. Cl.
`A 6LX 39/395
`C07K I4/47
`C40B 30/02
`C07K 6/8
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.015
`
`(2006.01)
`CI2N 5/00
`(2006.01)
`A6IP 9/02
`(2006.01)
`A6IP35/00
`(2006.01)
`A6IPI/00
`(2006.01)
`A6IP 7/06
`(2006.01)
`GOIN 33/566
`(52) U.S. Cl. ....... 424/133.1; 530/350:506/8:530/387.3;
`435/29: 435/375; 424/139.1; 436/501:530/387.9
`ABSTRACT
`
`(57)
`
`The present invention relates to compositions and methods of
`using those compositions for the diagnosis and treatment of
`immune related diseases.
`
`Merck Ex. 1045
`Page 1 of 131
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`Page 3 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 3 0f 60
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`US 2009/0258013 A1
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`Merck Ex. 1045
`Page 4 of 131
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`Oct. 15, 2009 Sheet 4 of 60
`
`US 2009/02580 13 A1
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`Merck Ex. 1045
`Page 5 of 131
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`Patent Application Publication
`
`Oct. 15, 2009 Sheet 5 of 60
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`US 2009/02580 13 A1
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`Page 6 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 6 of 60
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`US 2009/02580 13 A1
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`Merck Ex. 1045
`Page 7 of 131
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`

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`Page 8 of 131
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`Merck Ex. 1045
`Page 8 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 8 of 60
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`US 2009/02580 13 A1
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`Merck Ex. 1045
`Page 9 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 9 of 60
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`US 2009/02580 13 A1
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`Merck Ex. 1045
`Page 10 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 10 of 60
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`US 2009/02580 13 A1
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`Merck Ex. 1045
`Page 11 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 11 of 60
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`US 2009/02580 13 A1
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`O. 12
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`Merck Ex. 1045
`Page 12 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 12 of 60
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`US 2009/02580 13 A1
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`Merck Ex. 1045
`Page 13 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 13 of 60
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`US 2009/02580 13 A1
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`CHO-CD226
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`FIG. 7A
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`800
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`FIG. 7B
`
`Merck Ex. 1045
`Page 14 of 131
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`Oct. 15, 2009 Sheet 14 of 60
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`US 2009/0258013 A1
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`Baso
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`Eff 15h us
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`FIG. 8A
`
`Merck Ex. 1045
`Page 15 of 131
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`US 2009/02580 13 A1
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`10000
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`Merck Ex. 1045
`Page 16 of 131
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`Oct. 15, 2009 Sheet 16 of 60
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`US 2009/02580 13 A1
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`Merck Ex. 1045
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`Merck Ex. 1045
`Page 18 of 131
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`Merck Ex. 1045
`Page 19 of 131
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`US 2009/02580 13 A1
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`TGIT mRNA
`Fold Change
`
`
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`RA CD4 RO Treg NK DC
`FIG. 10B
`
`Merck Ex. 1045
`Page 20 of 131
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`Oct. 15, 2009 Sheet 20 of 60
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`OO CO Vr CN O
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`Merck Ex. 1045
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`Merck Ex. 1045
`Page 22 of 131
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`Oct. 15, 2009 Sheet 22 of 60
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`:
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`Merck Ex. 1045
`Page 23 of 131
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`Merck Ex. 1045
`Page 24 of 131
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`2x10^5
`-- TIG-
`2x10^5
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`FIG. 12B
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`
`0.6
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`O.8
`
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`Anti-Ragweed
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`TIGIT TIGIT, DANA isotypes
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`With Control
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`0.05
`
`Anti-CD40
`
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`
`F.G. 13A
`
`Merck Ex. 1045
`Page 25 of 131
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`Oct. 15, 2009 Sheet 25 of 60
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`US 2009/02580 13 A1
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`10000
`9 OOO
`8000
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`FIG. 13C
`
`Merck Ex. 1045
`Page 26 of 131
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`CCR6-Th1 (CXCR3+)
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`m
`
`FIG. 14
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`i 40
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`
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`
`Merck Ex. 1045
`Page 27 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 27 of 60
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`US 2009/02580 13 A1
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`
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`Merck Ex. 1045
`Page 28 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 28 of 60
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`US 2009/02580 13 A1
`
`20
`
`18
`16
`14
`12
`10
`
`3.0
`
`
`
`2.5
`
`2.0
`
`15
`
`0.5
`
`0.0
`
`TIGIT Expression in Mouse CIA Model
`
`ZZZZZZZ
`
`CD226 Expression in Mouse CIA Model
`
`2
`
`Day
`
`FIG. 16
`
`Merck Ex. 1045
`Page 29 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 29 of 60
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`US 2009/0258013 A1
`
`TGIT
`
`PVR
`
`
`
`
`
`CD226
`
`Dis
`Ctrl
`Baseline
`
`Dis
`Ctrl
`4 MOnth
`
`Dis
`Ctrl
`8 MOnth
`
`Dis
`Ctrl
`12 MOnth
`
`Dis
`Ctrl
`Baseline
`
`Dis
`Ctrl
`4 MOnth
`
`Dis
`Ctrl
`8 MOnth
`
`Dis
`Ctrl
`12 MOnth
`
`Dis
`Ctrl
`Baseline
`
`Dis
`Ctrl
`4 Month
`
`Dis
`Ctrl
`8 Month
`
`Dis
`Ctrl
`12 Month
`
`FIG. 17
`
`Merck Ex. 1045
`Page 30 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 30 of 60
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`US 2009/02580 13 A1
`
`TIGIT Expression
`
`1500
`
`d
`d 1000
`.9
`f
`SD
`O 500
`X
`
`O
`
`(N)
`
`
`
`3OOO
`
`-
`
`d
`a 2000
`O
`f
`f
`9)
`as 1000
`X
`
`500
`
`CD4
`
`O
`
`(D)
`
`(M)
`
`C
`(NNormal
`(C) Cancer
`(M) Metastasis
`(D Disease (Not Cancer)
`p < 0.0001 for comparison between
`normal and cancer or metastasis
`CD4 Expression
`Breast Tumors
`
`He
`
`<
`s
`
`2 S
`O. Z
`
`Cd
`ve
`
`v
`
`cN
`
`S.
`
`ve
`
`O
`
`.
`
`cN
`
`l
`N.
`
`.
`
`D
`
`C
`O
`
`N
`
`.
`
`FIG. 18A
`
`Cell Type
`
`O
`O
`
`;
`
`Merck Ex. 1045
`Page 31 of 131
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`Patent Application Publication
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`Oct. 15, 2009 Sheet 31 of 60
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`US 2009/02580 13 A1
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`TGIT
`
`1500
`
`1000
`
`O
`
`
`
`1000
`
`O
`
`FIG. 18B
`
`Merck Ex. 1045
`Page 32 of 131
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`

`Patent Application Publication
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`Oct. 15, 2009 Sheet 32 of 60
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`US 2009/02580 13 A1
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`PVR
`
`
`
`CD O OO
`
`600
`
`O
`
`200
`
`06-9/
`
`FIG. 18C
`
`Merck Ex. 1045
`Page 33 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 33 of 60
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`US 2009/02580 13 A1
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`CD226
`
`250
`
`200
`
`150
`
`1 OO
`
`
`
`
`
`FIG. 18D,
`
`Merck Ex. 1045
`Page 34 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 34 of 60
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`US 2009/02580 13 A1
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`FIG. 19A
`
`PVR
`
`PVR
`
`PVR
`
`Merck Ex. 1045
`Page 35 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 35 of 60
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`US 2009/0258013 A1
`
`Olsotype/IgG
`E TIGIT.Fc/IgG
`TGITFC/1OA7
`TIGIT.Fc/Anti-PVR
`
`
`
`3OOOO
`
`Ys
`
`9, 20000-
`5
`9 10000
`s
`
`O
`
`
`
`- 60000
`E
`Q-
`Sl
`5 40000
`9
`is 20000
`f
`
`O
`
`Olsotype
`80000 ETIGIT.Fc
`
`A.
`
`60000
`
`Sl
`C
`S 40000
`s
`D
`5 20000
`O
`
`
`
`1000
`
`800
`1N
`E
`S. 600
`3.
`C
`400
`
`200
`
`O
`
`100
`
`2n
`8
`s
`St 50
`>-
`
`FIG. 19D
`
`Merck Ex. 1045
`Page 36 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 36 of 60
`
`US 2009/02580 13 A1
`
`
`
`H–H H-007
`
`Merck Ex. 1045
`Page 37 of 131
`
`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 37 of 60
`
`US 2009/02580 13 A1
`
`N.
`CC
`C
`ve
`
`CD
`C.
`e
`r
`C
`
`C
`
`v
`
`D
`O
`5.
`
`N.
`s
`3.
`5
`2
`
`ce
`
`O
`t
`
`
`
`s
`
`A.
`
`c.
`co
`
`c.
`v .
`di Co
`
`c.
`N.
`Lo
`co
`O o o co
`E
`
`o
`O
`
`
`
`O
`s
`
`his
`
`n-
`
`% Proliferation
`
`Q
`VN
`GN
`g
`U
`
`O)
`c)
`s
`e
`t
`o
`H
`
`O
`D
`H
`CD
`
`d b d b d d did
`N.
`cd
`w
`cro
`n v
`% Suppression
`
`db
`
`VN
`GN
`9
`U
`
`Merck Ex. 1045
`Page 38 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 38 of 60
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`US 2009/02580 13 A1
`
`
`
`L-10
`
`FIG.22A-1
`
`I
`
`FIG
`
`Merck Ex. 1045
`Page 39 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 39 of 60
`
`US 2009/0258013 A1
`
`IL-12/23p40
`
`2000
`
`1500
`
`1000
`
`500
`
`2500
`
`2000
`
`1500
`
`1000
`
`500
`
`2000
`1500
`
`100 O
`
`
`
`FIG.22A-2
`
`C
`O
`C
`e
`
`i
`
`Merck Ex. 1045
`Page 40 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 40 of 60
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`US 2009/02580 13 A1
`
`FIG. 22A-3
`
`IL-12p70
`
`200
`
`150
`
`1OO
`
`50
`
`150
`
`1OO
`
`50
`
`600
`
`400
`
`200
`
`200
`
`
`
`150
`
`1OO
`
`50
`
`S
`e
`
`3. es
`5.
`
`s
`
`i
`
`Merck Ex. 1045
`Page 41 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 41 of 60
`
`US 2009/02580 13 A1
`
`
`
`8
`O
`O
`
`O
`O
`V
`
`
`
`CY)
`2
`O
`
`O
`H
`CD
`Hr
`CD
`O
`2n
`v O
`O2
`
`C
`O
`cy
`
`C
`O
`CN
`
`O
`O
`w
`
`O
`
`O
`
`H
`CD
`H
`CD
`O
`2n
`O
`O
`
`O O O O O O
`O CO O r
`CN
`ve
`
`O
`H
`CD
`H
`8.
`2n
`O
`2
`
`O
`3
`O
`
`5 3 s 3
`
`O
`O
`S
`C
`On 5
`is 2
`O3:
`
`Merck Ex. 1045
`Page 42 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 42 of 60
`
`US 2009/02580 13 A1
`
`TNFO-Matured
`
`2000
`
`1500
`
`LPS-Matured
`
`2000
`
`1500
`
`IL-6 pg/ml 1000
`
`pg/ml 1000
`
`500
`
`80
`
`isotype TIGIT-Fo
`TNFO-Matured
`
`isotype TIGIT-Fo
`LPS-Matured
`
`500
`
`100
`
`75
`
`pg/ml 50
`
`25
`
`60
`
`IL-12p70
`pg/ml 40
`
`20
`
`O
`
`
`
`IL-18 pg/ml
`
`isotype TIGIT-Fo
`TNFO-Matured
`
`isotype TIGIT-Fo
`LPS-Matured
`
`isotype TIGIT-Fo
`isotype TIGIT-Fo
`N- - - -
`FIG.22C
`
`Merck Ex. 1045
`Page 43 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 43 of 60
`
`US 2009/02580 13 A1
`
`isotype
`TGITFC
`FIG. 22D
`
`iSO - TGT TIGT.
`DANA
`
`
`
`12OO
`
`900
`TGFB (pg/ml) soo
`
`300
`
`iSO TIGIT
`
`
`
`s
`FIG. 23A
`
`5
`iso TIGIT
`
`30'
`iso TIGIT
`
`120'
`iso TIGIT
`
`
`
`FIG. 23C
`
`Active
`B-catenin
`
`Total
`B-catenin
`
`Merck Ex. 1045
`Page 44 of 131
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`Patent Application Publication
`
`Oct. 15, 2009 Sheet 44 of 60
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`US 2009/02580 13 A1
`
`EDMSO OERKK inhib
`S
`
`TIGIT-Fc isotype. DANA TIGIT-Fc-DANA
`
`isotype
`FIG. 24A
`
`
`
`OControl
`1OA7
`all 10 ;
`DaCD32
`
`Isotype
`FIG. 24B
`
`TGT. FC
`
`Merck Ex. 1045
`Page 45 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 45 of 60
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`US 2009/02580 13 A1
`
`1OO O O
`
`O
`
`a
`
`A.
`
`M
`
`MDDC
`
`TNFO-MDDC CD40L-MDDC
`
`2n
`S
`3.
`cN
`
`100
`
`75
`
`50
`25
`
`O
`
`iMDDC
`
`TNFO-MDDC
`
`()
`
`IL-10. KO
`
`()
`
`a 300
`S
`w
`8 200
`St
`.9
`H 100
`s
`
`W.
`
`O
`
`WT
`
`()
`()
`
`()
`
`
`
`300
`
`200
`
`1 OO
`
`O
`
`FIG. 27A
`
`H-
`
`O
`
`H
`
`O
`
`Merck Ex. 1045
`Page 46 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 46 of 60
`
`US 2009/02580 13 A1
`
`
`
`OCTOWI ------
`
`08
`
`ZITI
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`Merck Ex. 1045
`Page 47 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 47 of 60
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`US 2009/02580 13 A1
`
`7000
`
`- He Control
`6000 -O- TGT. FC
`- - - A - - - CTLA-4.1
`
`5000
`
`4000
`
`3000
`
`2000
`
`1000
`
`Fig. 27B
`
`O2
`
`KLH (ug/ml)
`
`2
`
`20
`
`400
`
`
`
`300
`
`200
`
`100
`
`FIG. 27c
`
`Merck Ex. 1045
`Page 48 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 48 of 60
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`US 2009/02580 13 A1
`
`20000
`
`16OOO
`
`12000
`
`8000
`
`FIG. 27D
`
`O.2
`
`KLH (ug/ml)
`
`2
`
`2O
`
`
`
`-H isotype
`-O- TGT. FC
`- - - A - - - CTLA-4.Fc
`
`IL-10.KO
`
`20OOO
`
`16000
`
`
`
`Yse 1 2 O O O
`
`8000
`
`4000
`
`FIG. 27E
`
`O.2
`
`KLH (ug/ml)
`
`2
`
`2O
`
`Merck Ex. 1045
`Page 49 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 49 of 60
`
`US 2009/02580 13 A1
`
`2000
`
`E 15OO
`S.
`St 1 OOO
`-
`Z
`
`500
`
`
`
`- 6000
`E
`s
`St 4000
`2
`- 2000
`
`e
`
`Olsotype
`TGT. FC
`CTLA4
`Normal
`
`
`
`
`
`IL-10. KO
`
`6OOO
`e
`S
`5 4000
`s
`
`2000
`
`O isotype
`is TIGIT.Fc
`CTLA4
`2 Normal
`
`YZZY
`
`O
`
`A.
`
`'...leaase
`
`FIG. 27G
`
`Merck Ex. 1045
`Page 50 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 50 of 60
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`US 2009/02580 13 A1
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`TIGITFC
`
`Isotype
`
`TGT. FC
`
`Isotype
`
`s
`
`TIGITFC
`
`Sotype
`
`TIGITFC
`
`isotype
`
`s
`
`isotype
`
`TIGITFC i
`TGT. FC i
`
`
`
`
`
`8
`
`8
`
`Merck Ex. 1045
`Page 51 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 51 of 60
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`US 2009/02580 13 A1
`
`TGT
`
`CTLA4
`
`
`
`s
`
`C
`
`10
`-20
`3
`CD 5 SS -50
`s 5 s -80
`S -110
`5
`g -140
`N1
`
`Merck Ex. 1045
`Page 52 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 52 of 60
`
`US 2009/02580 13 A1
`
`
`
`ºººOWN JIS —
`
`ºººOWN JIS
`
`
`
`
`
`
`
`ES-HO
`
`ºººOWN JIS —
`
`Cell Count
`
`Merck Ex. 1045
`Page 53 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 53 of 60
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`US 2009/02580 13 A1
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`
`
`
`
`
`
`
`
`
`
`ES-JO
`
`Cell Count
`
`Merck Ex. 1045
`Page 54 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 54 of 60
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`US 2009/02580 13 A1
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`
`
`Merck Ex. 1045
`Page 55 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 55 of 60
`
`US 2009/02580 13 A1
`
`
`
`C
`Y
`r
`O
`O
`
`CD226
`
`CD226
`
`Merck Ex. 1045
`Page 56 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 56 of 60
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`US 2009/02580 13 A1
`
`
`
`O
`
`w
`
`CN
`
`C
`
`
`
`(2
`C)
`b
`CD
`-
`CC
`Z
`4.
`E
`
`Merck Ex. 1045
`Page 57 of 131
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`

`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 57 of 60
`
`US 2009/02580 13 A1
`
`FIG. 29D
`
`12
`10
`
`mRNA
`Fold Change
`
`CD25
`CD25hi
`
`
`
`Activated
`
`Activated
`CD226
`
`FIG. 29E
`
`Merck Ex. 1045
`Page 58 of 131
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`

`Patent Application Publication
`
`Oct. 15, 2009 Sheet 58 of 60
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`US 2009/02580 13 A1
`
`o o o O O. C. co o cd
`O. O. O. O. O. O. O. O.
`O O. d O C C C Co
`tr V.
`co
`?o on O co cy
`CN.
`N. We was v
`
`Cell Proliferation (cpm)
`
`O
`N1
`H
`CD
`H
`
`H
`S
`
`O
`Y.
`H
`CD
`H
`
`(N
`wer
`
`CC
`H
`
`2n
`O
`H
`
`O o O O O O O
`O O O O O O
`O O C C O O
`Q Sp
`CO
`d
`V
`CN
`Cell Proliferation (cpm)
`
`
`
`H
`&
`
`O
`NZ
`H
`CD
`H
`
`Cell Proliferation (cpm)
`
`Merck Ex. 1045
`Page 59 of 131
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`Patent Application Publication
`
`Oct. 15, 2009 Sheet 59 of 60
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`US 2009/02580 13 A1
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`
`
`TGIT.KO
`
`IL-4
`
`FIG. 30B
`
`Merck Ex. 1045
`Page 60 of 131
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`
`Oct. 15, 2009 Sheet 60 of 60
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`US 2009/0258013 A1
`
`S
`IFNy (ng/ml)
`
`
`
`H
`s
`
`O
`N1.
`H
`CD
`H
`
`H
`s
`
`O
`Ng
`H
`CD
`
`O
`S
`
`CN
`
`O
`
`v-
`
`Od
`8
`
`ve
`
`O
`
`O
`
`O
`s
`
`C
`to
`9
`to
`TNFol (pg/ml)
`
`H.
`s
`
`O
`N.
`H
`O
`H
`
`d
`
`
`
`u?
`CN
`
`to
`
`c
`O
`
`O
`S.
`
`O
`O
`O
`S. S
`S
`IL-12p70 (pg/ml)
`
`H
`s
`
`O
`NZ
`CD
`H
`
`IL-2 (ng/ml)
`
`H
`S
`
`O
`N4
`H
`
`CY
`
`CD 8 S
`9 U
`
`H
`s
`
`O
`N4
`H
`CD
`H
`O
`
`H
`s
`
`O
`Na
`H
`CD
`
`Merck Ex. 1045
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`US 2009/02580 13 A1
`
`Oct. 15, 2009
`
`NOVELCOMPOSITIONS AND METHODS
`FOR THE TREATMENT OF IMMUNE
`RELATED DISEASES
`
`RELATED APPLICATIONS
`0001. This application claims priority under 35 U.S.C.
`119(e) to U.S. provisional application No. 61/123.530, filed
`Apr. 9, 2008, and to U.S. provisional application No. 61/194,
`271, filed Sep. 26, 2008, the contents of which are incorpo
`rated in their entirety herein by reference.
`
`FIELD OF THE INVENTION
`0002 The present invention relates to compositions and
`methods useful for the diagnosis and treatment of immune
`related diseases.
`
`BACKGROUND OF THE INVENTION
`0003. Immune related and inflammatory diseases are the
`manifestation or consequence of fairly complex, often mul
`tiple interconnected biological pathways which in normal
`physiology are critical to respond to insult or injury, initiate
`repair from insult or injury, and mount innate and acquired
`defense against foreign organisms. Disease or pathology
`occurs when these normal physiological pathways cause
`additional insult or injury either as directly related to the
`intensity of the response, as a consequence of abnormal regu
`lation or excessive stimulation, as a reaction to self, or as a
`combination of these.
`0004 Though the genesis of these diseases often involves
`multistep pathways and often multiple different biological
`systems/pathways, intervention at critical points in one or
`more of these pathways can have an ameliorative ortherapeu
`tic effect. Therapeutic intervention can occur by either
`antagonism of a detrimental process/pathway or stimulation
`of a beneficial process/pathway.
`0005. Many immune related diseases are known and have
`been extensively studied. Such diseases include immune
`mediated inflammatory diseases, non-immune-mediated
`inflammatory diseases, infectious diseases, immunodefi
`ciency diseases, neoplasia, etc.
`0006 T lymphocytes (T cells) are an important compo
`nent of a mammalian immune response. T cells recognize
`antigens which are associated with a self-molecule encoded
`by genes within the major histocompatibility complex
`(MHC). The antigen may be displayed together with MHC
`molecules on the Surface of antigen presenting cells, virus
`infected cells, cancer cells, grafts, etc. The T cell system
`eliminates these altered cells which pose a health threat to the
`host mammal. T cells include helper T cells and cytotoxic T
`cells. Helper T cells proliferate extensively following recog
`nition of an antigen-MHC complex on an antigen presenting
`cell. Helper T cells also secrete a variety of cytokines, i.e.,
`lymphokines, which play a central role in the activation of B
`cells, cytotoxic T cells and a variety of other cells which
`participate in the immune response. Another Subcategory of
`helper T cells are the follicular helper T cells (T,) (for
`review, see Vineusa et al., Nat. Rev. Immunol. 5: 853-865
`(2005)). Detectable by their characteristic expression of
`CXC-chemokine receptor 5 (Schaerli et al., J. Exp. Med. 192:
`1553-62 (2000)), these cells have been found to produce
`IL-10 and possibly IL-21. T, cells provide assistance to
`germinal-center B cells, particularly aiding the Survival and
`
`propagation of B cells and potently inducing antibody pro
`duction during coculture with B cells. They have also been
`implicated in tolerogenesis.
`10007 Regulatory T cells (T,) are a subset of helper T
`cells that play a critical role in inhibition of self-reactive
`immune responses and are often found in sites of chronic
`inflammation such as in tumor tissue (Wang, H.Y. & Wang, R.
`F., Curr Opin Immunol 19, 217-23 (2007)). T are defined
`phenotypically by high cell Surface expression of CD25,
`CLTA4, GITR, and neuropilin-1 (Read, S., Malmstrom, V. &
`Powrie, F.J Exp Med 192, 295-302 (2000); Sakaguchi, S., et
`al., JImmunol 155, 1151-64 (1995); Takahashi, T. et al., J Exp
`Med 192,303-10 (2000); McHugh, R. S. et al., Immunity 16,
`311-23 (2002); Bruder, D. et al., EurJ Immunol 34, 623-30
`(2004)), and are under the control of the transcription factor
`FOXP3 (Hori, S., Nomura, T. & Sakaguchi, S., Science 299,
`1057-61 (2003)).T. perform their suppressive function on
`activated T cells through contact-dependent mechanisms and
`cytokine production (Fehervari, Z. & Sakaguchi, Curr Opin
`Immunol 16, 203-8 (2004)). T,
`also modulate immune
`responses by direct interaction with ligands on dendritic cells
`(DC), such as CTLA4 interaction with B7 molecules on DC
`that elicits the induction of indoleamine 2,3-dioxygenase
`(IDO) (Fallarino, F. et al., Nat Immunol 4, 1206-12 (2003)),
`and CD40L ligation (Serra, P. et al., Immunity 19, 877-89
`(2003)). DCs are professional antigen-presenting cells
`capable of inducing immunity or tolerance against self or
`non-self antigens. DC-expanded T suppress alloreactivity
`responses in vitro (Yamazaki, S. et al., Proc Natl Acad Sci
`USA 103, 2758-63 (2006); Ahn, J. S. Krishnadas, D. K. &
`Agrawal, Int Immunol 19, 227-37 (2007)), and when adop
`tively transferred, appropriate T inhibited diabetes in
`NOD.scid mice (Tarbell, K. V. et al., J Exp Med 199, 1467-77
`(2004)) or experimentally induced asthma (Lewkowich, I. P.
`et al. J Exp Med 202, 1549-61 (2005)). Specific interactions
`of ligands on DC with T.
`can also abrogate their suppres
`sive function, Such as engagement of GITR in mice (Shimizu,
`J., et al., Nat Immunol 3, 135-42 (2002)), suggesting DC may
`have a pluralistic role in modulating T. function.
`0008. The molecules CTLA4 and GITR are representative
`of ligands defined within the CD28-B7 and TNF-superfami
`lies of co-stimulatory/-inhibitory molecules, respectively
`(Greenwald, R. J., et al., Annu Rev Immunol 23, 515-48
`(2005)). These molecules are high on T.
`but are also typi
`cally upregulated on activated T cells. In order to search for
`new co-stimulatory molecules expressed in T cells
`searches were performed to identify genes specifically
`expressed in T cells (Abbas, A. R. et al., Genes Immun 6.
`319-31 (2005)) that had both Ig domains and immunorecep
`tor tyrosine-based activation or inhibition (ITAM/ITIM)
`motifs. Through the intersection of these two genome-wide
`bioinformatics search strategies a novel cell Surface-bound
`protein with the protein encoding an IgV domain, a trans
`membrane domain, and two putative immunoreceptor
`tyrosine inhibitory motifs was identified (see US patent pub
`lication no. US20040121370, incorporated herein by refer
`ence). The protein designated TIGIT (for T-Cell-Ig and ITIM
`domain) was shown to be expressed on T cells-particularly
`T, and memory cell subsets—as well as NK cells. There is
`a need for new therapeutics and methods of treatment to
`address immune disorders, particularly autoimmune disor
`ders. Herein, Applicants identify TIGIT binding partners and
`provide new compositions, detection methods, and methods
`of treatment for immune disorders modulated by TIGIT inter
`
`reg
`
`Merck Ex. 1045
`Page 62 of 131
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`US 2009/02580 13 A1
`
`Oct. 15, 2009
`
`action with those binding partners and the elucidated TIGIT
`effects on T cell maturation and activity.
`
`SUMMARY OF THE INVENTION
`
`0009. The present invention concerns compositions and
`methods useful for the diagnosis and treatment of immune
`related disease in mammals, including humans. The present
`invention is based on the identification of proteins involved in
`the negative regulation of proliferation and function of certain
`types of immune cells. Immune related diseases can be
`treated by suppressing or enhancing the immune response.
`Molecules that enhance the immune response stimulate or
`potentiate the immune response to an antigen. Molecules
`which stimulate the immune response can be used therapeu
`tically where enhancement of the immune response would be
`beneficial. Alternatively, molecules that suppress the immune
`response attenuate or reduce the immune response to an anti
`gen (e.g., neutralizing antibodies) can be used therapeutically
`where attenuation of the immune response would be benefi
`cial (e.g., inflammation). Herein. Applicants demonstrate that
`TIGIT (for "T-Cell-Ig and ITIM domain”) protein specifi
`cally binds to poliovirus receptor (PVR, also known as CD
`155) and several other members of a newly elucidated protein
`family, and that this TIGIT-PVR interaction negatively regu
`lates T cell activation and proliferation. Accordingly, TIGIT
`polypeptides, agonists thereof, and antagonists thereof, as
`well as PVR polypeptides, agonists thereof and antagonists
`thereofare useful to prepare medicines and medicaments for
`the treatment of immune-related and inflammatory diseases.
`The invention also provides methods of treating immune
`related and inflammatory diseases and methods and compo
`sitions for detecting and assessing the status of immune
`related and inflammatory diseases.
`0010. In one embodiment, the invention provides an iso
`lated polypeptide comprising an amino acid sequence com
`prising one or more of the following amino acids: an alanine
`at amino acid position corresponding to amino acid position
`67 of human TIGIT, a glycine at an amino acid position
`corresponding to amino acid position 74 of human TIGIT, a
`proline at an amino acid position corresponding to amino acid
`position 114 of human TIGIT, and a glycine at an amino acid
`position corresponding to amino acid position 116 of human
`TIGIT. In one aspect, the polypeptide is not PVR, PVRL1,
`PVRL2, PVRL3, PVRL4, TIGIT, CD96, or CD226. In
`another aspect, the polypeptide further comprises one or more
`of an amino acid selected from valine, isoleucine, and leu
`cine at an amino acid position corresponding to amino acid
`position 54 of human TIGIT, an amino acid selected from
`serine and threonine at an amino acid position corresponding
`to amino acid position 55 of human TIGIT, a glutamine at an
`amino acid position corresponding to amino acid position 56
`of human TIGIT, a threonine at an amino acid position cor
`responding to amino acid position 112 of human TIGIT, and
`an amino acid selected from phenylalanine and tyrosine at an
`amino acid position corresponding to amino acid position 113
`of human TIGIT. In another aspect, the polypeptide further
`comprises one or more structural submotifs selected from the
`following:
`0011 a. an amino acid selected from valine and isoleu
`cine at amino acid position 54-an amino acid selected
`from serine and threonine at amino acid position 55-a
`glutamine at amino acid position 56;
`
`0012 b. analanine at position 67-any amino acid at each
`of amino acid positions 68-73-a glycine at amino acid
`position 74; and
`0013 c. a threonine at amino acid position 112-an
`amino acid selected from phenylalanine and tyrosine at
`amino acid position 113-a proline atamino acid position
`114-any amino acid at amino acid position 115-a glycine
`at amino acid position 116,
`wherein the numbering of the amino acid positions corre
`sponds to the amino acid positions of human TIGIT, although
`the absolute numbering of the amino acids in the polypeptide
`may differ.
`0014. In another embodiment, the invention provides a
`method of determining whether a test polypeptide is a mem
`ber of the TLP family of polypeptides comprising aligning the
`amino acid sequence of the test polypeptide with an amino
`acid sequence of one or more members of the TLP family of
`polypeptides and assessing the presence or absence in the test
`polypeptide amino acid sequence of one or more of analanine
`at amino acid position corresponding to amino acid position
`67 of human TIGIT, a glycine at an amino acid position
`corresponding to amino acid position 74 of human TIGIT, a
`prolineatanamino acid position corresponding to amino acid
`position 114 of human TIGIT, and a glycine at an amino acid
`position corresponding to amino acid position 116 of human
`TIGIT. In another embodiment, the invention provides a
`method for identifying one or more members of the TLP
`protein family by identifying proteins in one or more
`sequence databases whose amino acid sequences comprise at
`least one amino acid selected from an alanine at amino acid
`position corresponding to amino acid position 67 of human
`TIGIT, a glycine at an amino acid position corresponding to
`amino acid position 74 of human TIGIT, a proline at an amino
`acid position corresponding to amino acid position 114 of
`human TIGIT, and a glycine at an amino acid position corre
`sponding to amino acid position 116 of human TIGIT.
`0015. In another embodiment, the invention provides an
`isolated agent that specifically interacts with one or more
`conserved or substantially conserved regions of TLP family
`members. In one aspect, the agent is an antagonist of the
`expression and/or activity of a TLP family member. In
`another aspect, the antagonist is selected from a small mol
`ec