Modulation of Checkpoint Expression in Tumor Microenvironment by Intratumoral Administration of a Novel TLR9 Agonist: Rationale for Combination Therapy

• Newimmunotherapyregimensinvolvingmultipleagentsrepresentahighlypromisingareaofcancerresearch;achallengeistoidentifyoptimalcombinations.

• Investigationsofcheckpointinhibitorssuchasipilimumab,nivolumab,andpembrolizumabhaveshownacorrelationbetweenclinicalactivityandcheckpointgeneexpression.

• PreclinicaldatahavedemonstratedthatintratumoraladministrationofIMO-2125,anovelTLR9agonistshowntoinduceinterferon-α,modulatesthetumormicroenvironmentandinducesantitumorimmuneresponses.

• ThecurrentstudywasundertakentoevaluatethetumormicroenvironmentfollowingintratumoralIMO-2125therapy.Thisevaluationincludesanalysisofcheckpointexpressionintreatedtumorsaswellasindistanttumors.

Intratumoral IMO-2125 induced potent antitumor activity and tumor checkpoint expression compared to subcutaneous administration in an A20 lymphoma model

CT26 colon carcinoma model (single injection, multiple dose levels)

B16 melanoma model

A20 lymphoma model

iderapharma.com

Wayne Jiang, Daqing Wang, Fugang Zhu, Lakshmi Bhagat, Jillian DiMuzio and Sudhir AgrawalIdera Pharmaceuticals, Cambridge, MA 02139

CT26 colon carcinoma model

T

BALB/c, female (n = 10)

Parameters evaluated

• Induction of tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment

• Tumor volume

• Checkpoint gene expression in tumor nodules

Placebo

C57BL/6, female, (n = 9)

BALB/c, female, (n = 10)

BALB/c, female, (n = 5)

intratumoralsubcutaneous

IMO-2125, i.t.

IMO-2125, s.c.

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

900

600

300

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2000

1000

1500

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2500

2000

1500

1000

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

2000

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1.2

1.4

0.8

0.4

1

0.6

0.2

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

12

10

6

8

2

4

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

2.5

1.5

3

2

0.5

1

00 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Fold

cha

nge

3.5

4

2.5

1.5

3

2

0.5

1

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Tum

or v

olum

e, m

m3

2500

1600

800

2000

1200

400

0

0 7 14 21

Placebo IMO-2125 i.t.

Days

IMO-2125 s.c.

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSIONCHANGES IN TUMOR VOLUME AND TIL INFILTRATION

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

PBS IMO-2125

IDO

Tubulin

300

200

100

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

450

300

150

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

Day 1 8

Treatment IMO-2125, 2.5 and 12.5 mg/kg, i.t.

10

Tumor implant 3 x 106 cells

Tumor implant 3 x 106 cells

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t. or s.c.

210 10 14

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t.

210 10 14

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

220 9 13 15

BALB/c, female, (n = 9)

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

270 9 13 15

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Tumor implant 1 x 106 cells

Tumor implant 3 x 106 cells

Tumor implant 2 x 106 cells

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Distant tumor

Treated tumor Distant tumorIDO1 PDL1 CEACAM1 OX40

CD3+ TILs

Treated tumor

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

Two weeks post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

IDO1

PDL1 LAG3

CTLA4

TIM3

0

3

6

9

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3 PDL1 LAG3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3

PDL1

LAG3

CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

0.0

1.5

3.0

4.5

IDO

1 TIM

3

PDL1

LAG3

CTLA4

Fold

incr

ease

ove

r pla

cebo

s.c i.t.

IDO1

PDL1

LAG3 CTLA4 TIM3

0

3

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3

CTLA4

0

2

4

8

Fold

incr

ease

com

pare

d to

pla

cebo

TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

T

BALB/c, female (n = 10)

Parameters evaluated

• Induction of tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment

• Tumor volume

• Checkpoint gene expression in tumor nodules

Placebo

C57BL/6, female, (n = 9)

BALB/c, female, (n = 10)

BALB/c, female, (n = 5)

intratumoralsubcutaneous

IMO-2125, i.t.

IMO-2125, s.c.

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

900

600

300

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2000

1000

1500

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2500

2000

1500

1000

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

2000

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1.2

1.4

0.8

0.4

1

0.6

0.2

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

12

10

6

8

2

4

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

2.5

1.5

3

2

0.5

1

00 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Fold

cha

nge

3.5

4

2.5

1.5

3

2

0.5

1

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Tum

or v

olum

e, m

m3

2500

1600

800

2000

1200

400

0

0 7 14 21

Placebo IMO-2125 i.t.

Days

IMO-2125 s.c.

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSIONCHANGES IN TUMOR VOLUME AND TIL INFILTRATION

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

PBS IMO-2125

IDO

Tubulin

300

200

100

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

450

300

150

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

Day 1 8

Treatment IMO-2125, 2.5 and 12.5 mg/kg, i.t.

10

Tumor implant 3 x 106 cells

Tumor implant 3 x 106 cells

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t. or s.c.

210 10 14

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t.

210 10 14

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

220 9 13 15

BALB/c, female, (n = 9)

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

270 9 13 15

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Tumor implant 1 x 106 cells

Tumor implant 3 x 106 cells

Tumor implant 2 x 106 cells

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Distant tumor

Treated tumor Distant tumorIDO1 PDL1 CEACAM1 OX40

CD3+ TILs

Treated tumor

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

Two weeks post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

IDO1

PDL1 LAG3

CTLA4

TIM3

0

3

6

9

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3 PDL1 LAG3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3

PDL1

LAG3

CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

0.0

1.5

3.0

4.5

IDO

1 TIM

3

PDL1

LAG3

CTLA4

Fold

incr

ease

ove

r pla

cebo

s.c i.t.

IDO1

PDL1

LAG3 CTLA4 TIM3

0

3

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3

CTLA4

0

2

4

8

Fold

incr

ease

com

pare

d to

pla

cebo

TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

T

BALB/c, female (n = 10)

Parameters evaluated

• Induction of tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment

• Tumor volume

• Checkpoint gene expression in tumor nodules

Placebo

C57BL/6, female, (n = 9)

BALB/c, female, (n = 10)

BALB/c, female, (n = 5)

intratumoralsubcutaneous

IMO-2125, i.t.

IMO-2125, s.c.

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

900

600

300

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2000

1000

1500

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2500

2000

1500

1000

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

2000

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1.2

1.4

0.8

0.4

1

0.6

0.2

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

12

10

6

8

2

4

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

2.5

1.5

3

2

0.5

1

00 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Fold

cha

nge

3.5

4

2.5

1.5

3

2

0.5

1

0Fo

ld c

hang

e0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Tum

or v

olum

e, m

m3

2500

1600

800

2000

1200

400

0

0 7 14 21

Placebo IMO-2125 i.t.

Days

IMO-2125 s.c.

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSIONCHANGES IN TUMOR VOLUME AND TIL INFILTRATION

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

PBS IMO-2125

IDO

Tubulin

300

200

100

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

450

300

150

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

Day 1 8

Treatment IMO-2125, 2.5 and 12.5 mg/kg, i.t.

10

Tumor implant 3 x 106 cells

Tumor implant 3 x 106 cells

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t. or s.c.

210 10 14

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t.

210 10 14

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

220 9 13 15

BALB/c, female, (n = 9)

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

270 9 13 15

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Tumor implant 1 x 106 cells

Tumor implant 3 x 106 cells

Tumor implant 2 x 106 cells

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Distant tumor

Treated tumor Distant tumorIDO1 PDL1 CEACAM1 OX40

CD3+ TILs

Treated tumor

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

Two weeks post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

IDO1

PDL1 LAG3

CTLA4

TIM3

0

3

6

9

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3 PDL1 LAG3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3

PDL1

LAG3

CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

0.0

1.5

3.0

4.5

IDO

1 TIM

3

PDL1

LAG3

CTLA4

Fold

incr

ease

ove

r pla

cebo

s.c i.t.

IDO1

PDL1

LAG3 CTLA4 TIM3

0

3

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3

CTLA4

0

2

4

8

Fold

incr

ease

com

pare

d to

pla

cebo

TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

T

BALB/c, female (n = 10)

Parameters evaluated

• Induction of tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment

• Tumor volume

• Checkpoint gene expression in tumor nodules

Placebo

C57BL/6, female, (n = 9)

BALB/c, female, (n = 10)

BALB/c, female, (n = 5)

intratumoralsubcutaneous

IMO-2125, i.t.

IMO-2125, s.c.

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

900

600

300

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2000

1000

1500

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2500

2000

1500

1000

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

2000

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1.2

1.4

0.8

0.4

1

0.6

0.2

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

12

10

6

8

2

4

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

2.5

1.5

3

2

0.5

1

00 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Fold

cha

nge

3.5

4

2.5

1.5

3

2

0.5

1

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Tum

or v

olum

e, m

m3

2500

1600

800

2000

1200

400

0

0 7 14 21

Placebo IMO-2125 i.t.

Days

IMO-2125 s.c.

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSIONCHANGES IN TUMOR VOLUME AND TIL INFILTRATION

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

PBS IMO-2125

IDO

Tubulin

300

200

100

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

450

300

150

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

Day 1 8

Treatment IMO-2125, 2.5 and 12.5 mg/kg, i.t.

10

Tumor implant 3 x 106 cells

Tumor implant 3 x 106 cells

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t. or s.c.

210 10 14

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t.

210 10 14

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

220 9 13 15

BALB/c, female, (n = 9)

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

270 9 13 15

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Tumor implant 1 x 106 cells

Tumor implant 3 x 106 cells

Tumor implant 2 x 106 cells

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Distant tumor

Treated tumor Distant tumorIDO1 PDL1 CEACAM1 OX40

CD3+ TILs

Treated tumor

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

Two weeks post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

IDO1

PDL1 LAG3

CTLA4

TIM3

0

3

6

9

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3 PDL1 LAG3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3

PDL1

LAG3

CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

0.0

1.5

3.0

4.5

IDO

1 TIM

3

PDL1

LAG3

CTLA4

Fold

incr

ease

ove

r pla

cebo

s.c i.t.

IDO1

PDL1

LAG3 CTLA4 TIM3

0

3

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3

CTLA4

0

2

4

8

Fold

incr

ease

com

pare

d to

pla

cebo

TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

T

BALB/c, female (n = 10)

Parameters evaluated

• Induction of tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment

• Tumor volume

• Checkpoint gene expression in tumor nodules

Placebo

C57BL/6, female, (n = 9)

BALB/c, female, (n = 10)

BALB/c, female, (n = 5)

intratumoralsubcutaneous

IMO-2125, i.t.

IMO-2125, s.c.

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

900

600

300

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2000

1000

1500

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

2500

2000

1500

1000

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

2000

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1500

1000

500

0

Tum

or v

olum

e, m

m3

Placebo IM

O-2

125

1.2

1.4

0.8

0.4

1

0.6

0.2

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

12

10

6

8

2

4

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

2.5

1.5

3

2

0.5

1

00 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Fold

cha

nge

3.5

4

2.5

1.5

3

2

0.5

1

0

Fold

cha

nge

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA,mg/kg

Tum

or v

olum

e, m

m3

2500

1600

800

2000

1200

400

0

0 7 14 21

Placebo IMO-2125 i.t.

Days

IMO-2125 s.c.

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSION

CHANGES IN CHECKPOINT GENE EXPRESSIONCHANGES IN TUMOR VOLUME AND TIL INFILTRATION

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

CHANGES IN TUMOR VOLUME

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

STUDY DESIGN

PBS IMO-2125

IDO

Tubulin

300

200

100

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

450

300

150

0

Tum

or v

olum

e, m

m3

0 2.5 12.5 12.5

IMO-2125, mg/kg

Control DNA, mg/kg

Day 1 8

Treatment IMO-2125, 2.5 and 12.5 mg/kg, i.t.

10

Tumor implant 3 x 106 cells

Tumor implant 3 x 106 cells

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t. or s.c.

210 10 14

Day 8 12

Treatment IMO-2125, 2.5 mg/kg, i.t.

210 10 14

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

220 9 13 15

BALB/c, female, (n = 9)

Day 7 11

Treatment IMO-2125, 2.5 mg/kg, i.t.

270 9 13 15

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Tumor implant 1 x 106 cells

Tumor implant 3 x 106 cells

Tumor implant 2 x 106 cells

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Parameters evaluated

• Tumor volume

• Checkpoint gene expression in tumor nodules

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Treated tumor Distant tumor

Treated tumor Distant tumor Distant tumor

Treated tumor Distant tumorIDO1 PDL1 CEACAM1 OX40

CD3+ TILs

Treated tumor

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

One week post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

Two weeks post last dosing, tumor samples were collected and analyzed for checkpoint expression by qPCR. Data represented average fold change in gene expression of selected checkpoints compared to placebo treated tumor.

IDO1

PDL1 LAG3

CTLA4

TIM3

0

3

6

9

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3 PDL1 LAG3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

TIM3

PDL1

LAG3

CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3 CTLA4

0

2

4

6

8

Fold

incr

ease

com

pare

d to

pla

cebo

0.0

1.5

3.0

4.5

IDO

1 TIM

3

PDL1

LAG3

CTLA4

Fold

incr

ease

ove

r pla

cebo

s.c i.t.

IDO1

PDL1

LAG3 CTLA4 TIM3

0

3

12

Fold

incr

ease

com

pare

d to

pla

cebo

IDO1

PDL1 LAG3 TIM3

CTLA4

0

2

4

8

Fold

incr

ease

com

pare

d to

pla

cebo

TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

TUMOR TUMOR

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

Treated tumor

Distant tumor

• IntratumoralIMO-2125stimulatedpotentantitumoractivityinpreclinicalmodelsoflymphoma,coloncarcinomaandmelanoma.

- TreatmentledtoincreasesinCD3+TILs.

• IntratumoralIMO-2125inducedasystemicantitumoreffect.

- Treatmentledtodecreasesintumorvolumeintreatedanddistanttumors.

• IntratumoralIMO-2125modulatedimmunecheckpointgeneexpression,includingIDO1,PDL1,TIM3,LAG3andCTLA4,inbothtreatedanddistanttumornodules.

• Together,thesedatashowedthatintratumoralIMO-2125sensitizedthetumormicroenvironmentforpotentialcombinatorialeffectswithvariouscheckpointinhibitors.

• PlanningforaclinicaltrialofintratumoralIMO-2125incombinationwithipilimumab,ananti-CTLA4monoclonalantibody,inpatientswithmetastaticmelanomaiscurrentlyunderwaywithstudyinitiationexpectedin4Q2015.

Single and multiple intratumoral IMO-2125 doses induced potent antitumor activity and checkpoint gene expression

METHODS AND RESULTS

CONCLUSIONS

INTRODUCTION