A Selective Inhibitor of Endosomal Toll -Like Receptors, IMO ...270 9 IMO-8400 Inhibits Expression...

A Selective Inhibitor of Endosomal Toll-Like A Selective Inhibitor of Endosomal Toll Like Receptors, IMO-8400, Suppresses Activation of

Multiple Cytokines, Th17 Response and I fl A ti tiInflammasome Activation

Presented at American College of Rheumatology/Association of Rheumatology Health Professionals (ACR/ARHP) Annual MeetingHealth Professionals (ACR/ARHP) Annual Meeting

November 9-14, 2012, Washington, D.C Abstract 1068

© 2012, Idera Pharmaceuticals www.iderapharma.com© 2011, Idera Pharmaceuticals www.iderapharma.com© 2012, Idera Pharmaceuticals www.iderapharma.com 1

IntroductionIntroduction• Idera’s therapeutic approach for the treatment of autoimmune diseases is to inhibit inductionIdera s therapeutic approach for the treatment of autoimmune diseases is to inhibit induction

of disease associated cytokines and signaling cascades by modulating activation of Toll-like Receptors (TLRs) 7, 8 and 9

• We have selected Psoriasis as disease model to evaluate the efficacy of our approach• Psoriasis is a chronic inflammatory skin disease primarily associated with a Th1 and Th17

response. The activation of inflammasomes and subsequent release of IL-1β has also been shown to contribute to disease development

• Current therapies for psoriasis include oral and injectable drugs such as Retinoids,Current therapies for psoriasis include oral and injectable drugs such as Retinoids, Methotrexate, Cyclosporine, Hydorxyurea and Biologics such as Amevive (anti-CD2 protein), Enbrel (anti-TNFα inhibitor), Remicade (anti-TNFα antibody), Stelara (anti-IL-12/23 antibody), Humira (anti-TNFα antibody) and Simponi (anti-TNFα antibody)

• We have developed novel TLR antagonists that block TLR7 8 and 9 signaling These• We have developed novel TLR antagonists that block TLR7, 8 and 9 signaling. These antagonists improve disease outcome in autoimmune disease models by blocking interaction of immune complexes carrying self nucleic acids with TLRs, thereby preventing induction of disease associated cytokines and signaling cascades

O f• IMO-3100, an antagonist of TLR7 and 9 is currently in Phase 2 clinical trial in patients with moderate to severe psoriasis

• IMO-8400, an antagonist of TLR7, 8 and 9, is in clinical development for the treatment of lupus

© 2012, Idera Pharmaceuticals www.iderapharma.com

p• In this study, we evaluated the ability of IMO-8400 to control disease development in mouse

models of psoriasis

2

IMO-8400 Inhibits Induction of Cytokines in NHP Mediated by TLR7, 8 or 9

0

25

e-do

se

0

25e-

dose

0

25

e-do

se

0

25

e-do

se

TLR7 TLR8 TLR9 TLR4

TNF-αIL-6IL-12IL-1β-50

-25

ange

from

pre

-50

-25

ange

from

pre

-50

-25

hang

e fr

om p

re

-50

-25

ange

from

pre

IFN-αIP-10

-100

-75

48 hr post-IMO-8400

% C

h

-100

-75

48 hr post-IMO-8400d i i t ti

% C

h

-100

-75

48 hr post-IMO-8400

% C

h

-100

-75

48 hr post-IMO-8400d i i t ti

% C

h

administration administration administration administration

Cynomolgus macaques (N=4) were injected sc with IMO-8400 at a dose of 1.5 mg/kgBlood was collected prior to IMO 8400 administration (pre dose) and 48hr post dosing

© 2012, Idera Pharmaceuticals www.iderapharma.com 3

Blood was collected prior to IMO-8400 administration (pre-dose) and 48hr post dosingPBMCs were isolated and cultured with agonist of TLR4, 7, 8 or 9Cytokine measurements in supernatants were carried out by multiplex or ELISA assay

TLR7, 8 and 9 in PsoriasisTLR7, 8 and 9 in Psoriasis

Injury or infection Psoriasis

TLR7TLR8 TLR9

IL 21 IL 22IL 12 IL 17Neutrophil

Keratinocyte Peptides

DNA/RNA

Th1 or Th17

IL-21, IL-22IL-23, IL-6

IL-12, IL-17, IFN-α, X

Peptides

Immunecomplexes

cells

TLR7, 8 and 9

complexes

Antagonist


Dendritic Cell

Experimental models of PsoriasisExperimental models of Psoriasis

Day 0 2 3 4 6 9 12 14 15 18

C57BL/6 miceear IL-23, 0.5 μg, i.d. once every 2 day

TerminateIMO-8400, s.c. Parameters:HistologyEar thickness

Experimental groups (N = 7)IMO-8400, 2.5 mg/kg Ear thicknessIMO-8400, 5 mg/kg

IMO-8400,15 mg/kg

Control Oligo, 15 mg/kg

PBS

Day 0 1 2 3 4 5 6

C57BL/6 micedorsal skin

TerminateIL-23, 1 μg, i.d. IMO-8400, s.c. Parameters:HistologyExperimental groups (N = 8)


gySkin mRNA Skin proteinSerum cytokine

IMO-8400, 15 mg/kg (300 µg)

PBS

IMO-8400 Suppresses IL-23-Induced Ear Thickness IncreaseIMO-8400 Suppresses IL-23-Induced Ear Thickness Increase

PBS IMO-8400Naive Control

HE stain, Magnification x 100Epidermal hyperplasia Inflammatory cell infiltration

m

450

IMO-8400: 15mg/kg

thic

knes

s,μm

390

420

**

Ear

PBS 2.5 mg/kg 5 mg/kg 15 mg/kg Control 360

IMO 840015 mg/kg


IMO-8400

*, P < 0.05 vs PBS group by two-tailed Student’s t test

IMO-8400 Inhibits Dorsal Skin Lesions Induced by IL-23IMO-8400 Inhibits Dorsal Skin Lesions Induced by IL-23

Naive PBS IMO-8400Naive PBS IMO 8400

**

HE stain, Magnification x 200Inflammatory cell infiltrationEpidermal hyperplasia * Abscess


IMO-8400 Inhibits Cytokine ProductionIMO-8400 Inhibits Cytokine Production

130330 IL-12IL-6

L-12

, pg/

ml

90

110

IL-6

, pg/

ml

110

220 * *

I

Naive PBS IMO-840070

90

Naive PBS IMO-84000

110

Naive PBS IMO 8400Naive PBS IMO 8400


*, P < 0.05 vs PBS group by two-tailed Student’s t test; Serum levels of cytokines were measured by ELISA

IMO-8400 Inhibits Expression of Multiple CytokinesIMO-8400 Inhibits Expression of Multiple Cytokines

IL 6 IFN γ

RQ 60

90Q

80

120 IL-6 IFN-γ

IFN

- γ, R

30

IL-6

, RQ

40

*

Naive PBS IMO-84000

Naive PBS IMO-84000

*

150 36IL-17A IL-17F

A, R

Q 100

F, R

Q

18

27* *

IL-1

7A

50 IL-1

7F

9

18


*, P < 0.05 vs PBS group by two-tailed Student’s t testNaive PBS IMO-8400

0Naive PBS IMO-8400

0

IL 21

IMO-8400 Inhibits Expression of Multiple CytokinesIMO-8400 Inhibits Expression of Multiple Cytokines

IL 22

Q

3000

4000

Q

44

66 IL-21 IL-22

IL-2

2, R

Q

1000

2000

IL-2

1, R

Q

22 **

Naive PBS IMO-84000

1 9

Naive PBS IMO-84000

IL-23 p19

9, R

Q

1.6

1.9

IL-2

3 p1

9

1.0

1.3

*


Naive PBS IMO-84000.7

10*, P < 0.05 vs PBS group by two-tailed Student’s t test; mRNA levels of cytokines in skin were measured by qPCR

IMO-8400 Does Not Affect Th2 CytokinesIMO-8400 Does Not Affect Th2 Cytokines

15

20

6

8IL-4 IL-10

IL-4

, RQ

5

10

IL-1

0, R

Q

2

4

Naive PBS IMO-84000

5

Naive PBS IMO-84000

2


mRNA levels of cytokines in skin were measured by qPCR

270 9

IMO-8400 Inhibits Expression of Anti-Microbial Peptides IMO-8400 Inhibits Expression of Anti-Microbial Peptides

DEFB4 S100A7a4,

RQ 180

7a, R

Q 6

DEF

B

90 S100

A7

3

*

*

Naive PBS IMO-84000

1.4

Naive PBS IMO-84000

S100A4 4 LL-37

0A4,

RQ 1.2

* -37,

RQ

2

3

S100

0 8

1.0 LL-

0

1


Naive PBS IMO-84000.8

12

* , P < 0.05 vs PBS group by two-tailed Student’s t test; mRNA levels of anti-microbial peptides in skin were measured by qPCR

Naive PBS IMO-84000

Inflammasome Pathway in PsoriasisInflammasome Pathway in Psoriasis

1st signal 2nd signal

Endosome9R7

ATPATP

RNADNA

TLR

8

TLR

9

TLR

MyD88MyD88 MyD88

NLRP3

CARD ASCPro-Caspase-1

Nucleus

NF-κB

pro IL-1β IL-1β

Caspase-1

psoriasisPro-inflammatory

cytokines


9 4

IMO-8400 Inhibits Inflammasome ActivationIMO-8400 Inhibits Inflammasome ActivationNLRP3 AIM2

3, R

Q 6

RQ

3

**

NLR

P3

3

AIM

2,

1

2*

21039

Naive PBS IMO-84000

Naive PBS IMO-84000

IL-1β IL-1β protein

mg

prot

ein

140

β, R

Q 26

IL-1

β, p

g/

70IL-1

β

13*

*


Naive PBS IMO-84000

Naive PBS IMO-84000

14

* , P < 0.05 vs PBS group by two-tailed Student’s t test; mRNA expression levels of inflammasome in skin were measured by qPCR;IL-1β levels in skin were measured by ELISA.

ConclusionConclusion

• IMO-8400, an antagonist of TLR7, 8 and 9 exerts a therapeutic effect in mouse models of psoriasis induced by IL-23• A li t di th l• Ameliorates disease pathology• Inhibits expression of Th1 cytokines such as IL-12, IL-6 and IFN-γ• Inhibits expression of genes associated with the IL-23/Th17 axis• Does not affect expression of Th2 cytokines IL-4 and IL-10• Inhibits expression of psoriasis-associated anti-microbial peptides such as

DEFB4, S100A4 and S100A7a• Inhibits inflammasome activation

• IMO-3100, a TLR7 and 9 antagonist in Phase II clinical trial for the gtreatment of moderate to severe psoriasis is equally effective as IMO-8400 in IL-23- induced psoriasis model


SummarySummary

• Antagonist blocks TLR activation thereby affecting the signaling cascade that controls the expression of multiple cytokines

• Inhibition of TLR activation ultimately results in the simultaneous blockadeInhibition of TLR activation ultimately results in the simultaneous blockade of many cellular events

• Blockade of TLR activation is not expected to perturb triggering of downstream signals mediated by other cellular eventsdownstream signals mediated by other cellular events

• Mechanism of action of the antagonist is significantly different from that of immunosuppressants or monoclonal antibodies utilized for the treatment of psoriasis and other autoimmune diseasesof psoriasis and other autoimmune diseases


A Selective Inhibitor of Endosomal Toll -Like Receptors, IMO ...270 9 IMO-8400 Inhibits Expression...

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