Tilsotolimod/aPD-1

PBS Tilsotolimod IDO-1i aPD-1

Tilsotolimod/IDO-1i IDO-1i/aPD-1 Triple

• Althoughcheckpointinhibitortherapyhasrevolutionizedcancertherapy,manypatientsremainrefractoryandfailtoachievedurableresponses

• Thissuboptimaloutcomeisduetomultiplemechanismsthatrangefromlowimmunogenicitytoinadequategenerationoractivationoftumor-specificTcells

• Combinationtherapiestargetingmultiplecheckpointsarelikelytoovercometheinadequateantitumorimmuneresponsesbyasingleagent

• Intratumoralinjectionoftilsotolimod(IMO-2125),aTLR9agonist,activatesdendriticcellstopresenttumorspecificantigensandenhancesT-lymphocyteinfiltrationwithininjectedtumors

• Intratumoraltilsotolimodtherapycreatesafavorabletumormicroenvironmentandthereforeenhancesantitumoractivitybysynergizingwithcheckpointinhibitortherapytoelicitaproductiveimmuneresponseincancerpatients

• Inthisstudy,weevaluatedacombinationimmunotherapyregimenthatinvolvesanimmuneactivator,tilsotolimod,andcheckpointinhibitorstargetingtwonegativeimmuneregulators,IDO-1andPD-1,toeliminatelargetumorsinsyngeneictumormodels

Graphic representation of intratumoral mechanism of action of tilsotolimod

EvrenKocabasArgon,FugangZhu,SudhirAgrawal,JonathanYingling,andDaqingWang

IderaPharmaceuticals,Inc.,Cambridge,MA,andExton,PA

Triplecombinationoftilsotolimod(IMO-2125),epacadostat,andanti-PD-1antibodydemonstratesmaximalantitumorefficacyanderadicateslargeestablishedtumorsinpreclinicalmodels

IderaPharma.com

Intratumoral tilsotolimod in combination with epacadostat and anti-PD-1 mAb exhibited maximal antitumor activity

Redline:completetumorregression

IntratumoraltilsotolimodtherapyenhancesinfiltrationofdendriticcellsandCD8+Tcellsinthetumormicroenvironment

Studydesigntoevaluateintratumoraltilsotolimodfortreatmentoflargeestablishedtumors

BALB/cmicereceivedasubcutaneousimplantof5x106murineCT26.CL25coloncarcinomacellsintheirrightflank(tumor1)andleftflank(tumor2).Treatmentwasinitiatedwhentumorswerefullyestablishedatday9(345±99mm3).Tilsotolimodat2.5mg/kg(50μgin100μL)wasgivenviaintratumoralinjectionintherighttumornodulesfivetimesondays9,12,14,16,and19.Tumornodulescollectedatday24wereanalyzedforcheckpointgeneexpressionbyqPCRandevidenceoftumor-infiltratinglymphocytes(TILs)byimmunohistochemicalstaining.

Abbreviations:i.t.,intratumoral;s.c.,subcutaneous.

Study design to evaluate intratumoral tilsotolimod in combination with IDO-1 inhibitor (epacadostat) and anti-PD-1 mAb in CT26.CL25 colon carcinoma tumor model

BALB/cmice(n=11-12pergroup)receivedasubcutaneousimplantof 2x106murineCT26.CL25coloncarcinomacellsintheirrightflank(tumor1)andleftflank(tumor2).Treatmentwasinitiatedonday10whentumorvolumereached200-400mm3.Tilsotolimod2.5mg/kg(50μgin100μLPBS)wasgivenviaintratumoralinjectionsintherighttumornodules2x/weekfor2weeks.IDO-1inhibitor(epacadostat,INCB24360,Cat.S7910,Selleckchem,100mg/kg)wasgivenviaintragastricadministration5x/weekfor2weeks.Anti-PD-1tumor(10mg/kg)wasgivenviaintraperitonealinjection3x/weekfor2weeks.

Abbreviations:i.g.,intragastric;i.t.,intratumoral;i.p.,intraperionteal.

Intratumoraltilsotolimodmediatedtumorregressioniscorrelatedwithtilsotolimod-mediatedupregulationofIDO-1andPD-L1inbothtreatedanddistanttumors

*Tumorgrowthinhibition,%=[1-(meanvolumeoftreatedtumors)/(meanvolumeofcontroltumors)]x100%.

StudydesigntoevaluatetilsotolimodincombinationwithaPD-1mAbandIDO-1inhibitor(epacadostat)in3LL-C75lungcarcinomaperitonealdisseminatedtumormodel

C57/BL6(female)mice(n=10pergroup)receivedanintraperitonealimplantof5x106murine3LL-C75Lewislungcarcinoma.Treatmentwasinitiatedonday8.Tilsotolimod2.5mg/kg(50μgin100μlPBS)wasgivenintraperitoneally2x/weekfor2weeks.IDO-1inhibitor(epacadostat,INCB24360,Cat.S7910,Selleckchem,75mg/kg)wasadministeredintraperitoneally5x/weekfor2weeks.Anti-PD-1tumor(10mg/kg)wasadministratedbyintraperitonealinjection3x/weekfor2weeks.Abbreviation:i.p.,intraperionteal.

• IntratumoraltilsotolimodtherapyincreasesTILinfiltrationandcheckpointexpression,creatingafavorabletumormicroenvironmenttoenhancetheantitumoractivityofcheckpointinhibitortherapies

• ThecombinationoftilsotolimodwithcheckpointinhibitorstargetingIDO-1andPD-1inducesmaximalantitumorefficacyanderadicateslargeestablishedtumorsinpreclinicalmodels

• Aphase1/2clinicaltrialinsubjectswithPD-1refractorymelanomahasshownencouragingresultsfortilsotolimodincombinationwithipilimumab;44%(4/9patients)achievedRECISTv1.1responsesinanti-PD-1refractorymelanomapatients(Diab,2017)

• Aphase3multicenterglobaltrialinthesamepopulationiscurrentlyenrolling(ILLUMINATE301,NCT03445533)

Reference: DiabA.Aphase1/2trialofintratumoral(i.t.)IMO-2125(IMO)incombinationwithcheckpointinhibitors(CPI)inPD-(L)1-refractorymelanoma.PresentedatESMO,September,2017.

Tumorvolumesatday24(3daysafterlasttreatment)

0

1 0 0 0

2 0 0 0

3 0 0 0

2 1 2 5P B S

Tum

orvo

lum

e,m

m3

ID O i a P D 1 ID O i2 1 2 5

a P D 12 1 2 5

ID O ia P D 1

ID O ia P D 12 1 2 5

0

1 0 0 0

2 0 0 0

3 0 0 0

2 1 2 5P B S

Tum

orvo

lum

e,m

m3

ID O i a P D 1 ID O i2 1 2 5

a P D 12 1 2 5

ID O ia P D 1

ID O ia P D 12 1 2 5

Treated tumor Distant tumor

73%

-6%

20%

84%

81%

19%

98%

58%

6%

19%

71% 69%

8%

88%

P<0.001

P=0.934

P=0.069

P<0.001

P=0.003

P=0.002

P<0.001

P=0.559

P=0.206

P<0.001

P=0.033

P=0.067

*Tumorgrowthinhibition,%=[1-(meanvolumeoftreatedtumors)/(meanvolumeofcontroltumors)]x100%.

PresentedattheAmericanAssociationforCancerResearchAnnualMeeting,April2018.

Tilsotolimod in combination with IDO-1 inhibitor (epacadostat) and anti-PD-1 mAb showed potent antitumor activity in 3LL-C75 lung carcinoma peritoneal disseminated tumor model

0 1 0 2 0 3 0 4 0 5 0 6 00

2 0

4 0

6 0

8 0

1 0 0

D a y s p o s t tu m o r im p la n ta t io n

Pe

rce

nt

su

rviv

al

P B S

IM O -2 1 2 5

ID O -1 i

a P D -1

IM O -2 1 2 5 + ID O -1 i

IM O -2 1 2 5 + a P D -1

ID O -1 i + a P D -1

T rip le

Daysposttumororimplantation

Perc

ents

urvi

val

Log-rank(Mantel–Cox)test: 1.PBSvstilsotolimod:P=0.023 2.PBSvsIDO-1inhibitor:P=0.941 3.PBSvsanti-PD-1mAb:P=0.706 4.TriplecombinationvsIDO-1inhibitor+anti-PD-1mAb:P=0.010

Tcellinfiltrationintumortissues

CD3IHCstainingx400,injectedtumor

• MoreTILswereobservedintumorstreatedwithtilsotolimodthanintumorstreatedwithPBS

• TILsdidnotincreasesignificantlyintumorstreatedwithIDO-1i,aPD-1,orboth

• SlightlymoreTILswereobservedintumorstreatedwithtilsotolimodplusIDO-1i,tilsotolimodplusaPD-1,orthetriplecombinationthanintumorstreatedwithtilsotolimodalone

IDO-1 PD-L1Tumor volume

PBS

Trea

ted

Dista

nt

- 2

0

2

4

6

8

IDO

-1 f

old

ch

an

ge

PBS

Trea

ted

Dista

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- 5

0

5

1 0

1 5

2 0

2 5

PD

-L1

fo

ld c

ha

ng

e

PBS

Trea

ted

Dista

nt

0

5 0 0

1 0 0 0

1 5 0 0

2 0 0 0

2 5 0 0

Tu

mo

r vo

lum

e,

mm

3

66%*

57%*

P=0.0002

P=0.004

P=0.01

P=0.16

P=0.003

P=0.02

T i l s o t o l i m o d

Tilsotolimod

Tilsotolimod

Groups (n=11-12/group):1. PBS2. Tilsotolimod 2.5 mg/kg, i.t.3. Epacadostat i.g.4. aPD-1 i.p.5. Tilsotolimod + Epacadostat i.g.6. Tilsotolimod + aPD-1 mAb i.p.7. Epacadostat i.g + aPD-1 i.p.8. Tilsotolimod + Epacadostat + aPD-1 mAb

Days 0

Dual solid tumors

CT26.CL25

CT26.CL25

Anti-PD-1 mAb i.p.

Tilsotolimod i.t.

Epacadostat i.g.

10 11 12 13 14 17 18 19 20 21

PBS Tilsotolimod

PBS Tilsotolimod

CD11c+ dendritic cells

CD8+ T cells

CD11c and CD8 IHC staining x 400, injected tumor

PBS Tilsotolimod IDO-1i aPD-1 Tilsotolimod/IDO-1i

Tilsotolimod/aPD-1

IDO-1i/aPD-1 Triple

0/12 2/12 1/11 0/11 2/12 1/11 1/12 10/11

Treated tumor

Distant tumor

0/12 2/12 1/11 0/11 3/12 2/11 0/12 6/11

Groups (n=10/group):1. PBS2. Tilsotolimod 2.5 mg/kg, i.p.3. Epacadostat 75 mg/kg, i.p.4. Anti-PD-1 10 mg/kg, i.p.5. Tilsotolimod + Epacadostat6. Tilsotolimod + Anti-PD-1 mAb7. Epacadostat + Anti-PD-1 mAb8. Tilsotolimod + Epacadostat + Anti-PD-1 mAb

Days 0

Peritoneal tumor

Tilsotolimod

Epacadostat

Anti-PD-1mAb

15 16 17 18 19 8 9 10 11 12

Groups (n=11-12/group):1. PBS2. Tilsotolimod 2.5 mg/kg, i.t.3. Epacadostat i.g.4. aPD-1 i.p.5. Tilsotolimod + Epacadostat i.g.6. Tilsotolimod + aPD-1 mAb i.p.7. Epacadostat i.g + aPD-1 i.p.8. Tilsotolimod + Epacadostat + aPD-1 mAb

Days 0

Dual solid tumors

CT26.CL25

CT26.CL25

Anti-PD-1 mAb i.p.

Tilsotolimod i.t.

Epacadostat i.g.

10 11 12 13 14 17 18 19 20 21

BACKGROUND

CONCLUSIONS

RESULTS