Triple combination of tilsotolimod (IMO-2125), epacadostat ......Tilsotolimod/aPD-1 PBS Tilsotolimod...
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Tilsotolimod/aPD-1 PBS Tilsotolimod IDO-1i aPD-1 Tilsotolimod/IDO-1i IDO-1i/aPD-1 Triple • Although checkpoint inhibitor therapy has revolutionized cancer therapy, many patients remain refractory and fail to achieve durable responses • This suboptimal outcome is due to multiple mechanisms that range from low immunogenicity to inadequate generation or activation of tumor- specific T cells • Combination therapies targeting multiple checkpoints are likely to overcome the inadequate antitumor immune responses by a single agent • Intratumoral injection of tilsotolimod (IMO-2125), a TLR 9 agonist, activates dendritic cells to present tumor specific antigens and enhances T-lymphocyte infiltration within injected tumors • Intratumoral tilsotolimod therapy creates a favorable tumor microenvironment and therefore enhances antitumor activity by synergizing with checkpoint inhibitor therapy to elicit a productive immune response in cancer patients • In this study, we evaluated a combination immunotherapy regimen that involves an immune activator, tilsotolimod, and checkpoint inhibitors targeting two negative immune regulators, IDO-1 and PD-1, to eliminate large tumors in syngeneic tumor models Graphic representation of intratumoral mechanism of action of tilsotolimod Evren Kocabas Argon, Fugang Zhu, Sudhir Agrawal, Jonathan Yingling, and Daqing Wang Idera Pharmaceuticals, Inc., Cambridge, MA, and Exton, PA Triple combination of tilsotolimod (IMO-2125), epacadostat, and anti-PD-1 antibody demonstrates maximal antitumor efficacy and eradicates large established tumors in preclinical models IderaPharma.com Intratumoral tilsotolimod in combination with epacadostat and anti-PD-1 mAb exhibited maximal antitumor activity Red line: complete tumor regression Intratumoral tilsotolimod therapy enhances infiltration of dendritic cells and CD8+ T cells in the tumor microenvironment Study design to evaluate intratumoral tilsotolimod for treatment of large established tumors BALB/c mice received a subcutaneous implant of 5 x 10 6 murine CT26.CL25 colon carcinoma cells in their right flank (tumor 1) and left flank (tumor 2). Treatment was initiated when tumors were fully established at day 9 (345 ± 99 mm 3 ). Tilsotolimod at 2.5 mg/kg (50 μg in 100 μ L) was given via intratumoral injection in the right tumor nodules five times on days 9, 12, 14, 16, and 19. Tumor nodules collected at day 24 were analyzed for checkpoint gene expression by qPCR and evidence of tumor-infiltrating lymphocytes (TILs) by immunohistochemical staining. 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/c mice (n=11-12 per group) received a subcutaneous implant of 2 x 10 6 murine CT26.CL25 colon carcinoma cells in their right flank (tumor 1) and left flank (tumor 2). Treatment was initiated on day 10 when tumor volume reached 200-400 mm 3 . Tilsotolimod 2.5 mg/kg (50 μg in 100 μ L PBS) was given via intratumoral injections in the right tumor nodules 2x/week for 2 weeks. IDO-1 inhibitor (epacadostat, INCB24360, Cat.S7910, Selleckchem, 100 mg/kg) was given via intragastric administration 5x/week for 2 weeks. Anti-PD-1 tumor (10 mg/kg) was given via intraperitoneal injection 3x/week for 2 weeks. Abbreviations: i.g., intragastric; i.t., intratumoral; i.p., intraperionteal. Intratumoral tilsotolimod mediated tumor regression is correlated with tilsotolimod-mediated upregulation of IDO-1 and PD-L1 in both treated and distant tumors *Tumor growth inhibition, % = [1 - (mean volume of treated tumors)/ (mean volume of control tumors)] x 100%. Study design to evaluate tilsotolimod in combination with aPD-1 mAb and IDO-1 inhibitor (epacadostat) in 3LL-C75 lung carcinoma peritoneal disseminated tumor model C57/BL6 (female) mice (n=10 per group) received an intraperitoneal implant of 5 x 10 6 murine 3LL-C75 Lewis lung carcinoma. Treatment was initiated on day 8. Tilsotolimod 2.5 mg/kg (50 μg in 100 μ l PBS) was given intraperitoneally 2x/week for 2 weeks. IDO-1 inhibitor (epacadostat, INCB24360, Cat.S7910, Selleckchem, 75 mg/kg) was administered intraperitoneally 5x/week for 2 weeks. Anti-PD-1 tumor (10 mg/kg) was administrated by intraperitoneal injection 3x/week for 2 weeks. Abbreviation: i.p., intraperionteal. • Intratumoral tilsotolimod therapy increases TIL infiltration and checkpoint expression, creating a favorable tumor microenvironment to enhance the antitumor activity of checkpoint inhibitor therapies • The combination of tilsotolimod with checkpoint inhibitors targeting IDO-1 and PD-1 induces maximal antitumor efficacy and eradicates large established tumors in preclinical models • A phase 1/2 clinical trial in subjects with PD-1 refractory melanoma has shown encouraging results for tilsotolimod in combination with ipilimumab; 44% (4/9 patients) achieved RECIST v1.1 responses in anti-PD-1 refractory melanoma patients (Diab, 2017) • A phase 3 multicenter global trial in the same population is currently enrolling (ILLUMINATE 301, NCT03445533) Reference: Diab A. A phase 1/2 trial of intratumoral (i.t.) IMO-2125 (IMO) in combination with checkpoint inhibitors (CPI) in PD-(L)1-refractory melanoma. Presented at ESMO, September, 2017. Tumor volumes at day 24 (3 days after last treatment) 0 1000 2000 3000 2125 PBS Tumor volume, mm 3 IDOi aPD1 IDOi 2125 aPD1 2125 IDOi aPD1 IDOi aPD1 2125 0 1000 2000 3000 2125 PBS Tumor volume, mm 3 IDOi aPD1 IDOi 2125 aPD1 2125 IDOi aPD1 IDOi aPD1 2125 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 *Tumor growth inhibition, % = [1 - (mean volume of treated tumors)/ (mean volume of control tumors)] x 100%. Presented at the American Association for Cancer Research Annual Meeting, April 2018. 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 10 20 30 40 50 60 0 20 40 60 80 100 Days post tumor implantation Percent survival PBS IDO-1i aPD-1 IMO-2125 + IDO-1i IMO-2125 + aPD-1 IDO-1i + aPD-1 Triple Days post tumor or implantation Percent survival Log-rank (Mantel–Cox) test: 1. PBS vs tilsotolimod: P =0.023 2. PBS vs IDO-1 inhibitor: P =0.941 3. PBS vs anti-PD-1 mAb: P =0.706 4. Triple combination vs IDO-1 inhibitor + anti-PD-1 mAb: P =0.010 T cell infiltration in tumor tissues CD3 IHC staining x 400, injected tumor • More TILs were observed in tumors treated with tilsotolimod than in tumors treated with PBS • TILs did not increase significantly in tumors treated with IDO-1i, aPD-1, or both • Slightly more TILs were observed in tumors treated with tilsotolimod plus IDO-1i, tilsotolimod plus aPD-1, or the triple combination than in tumors treated with tilsotolimod alone IDO-1 PD-L1 Tumor volume PBS Treated Distant -2 0 2 4 6 8 IDO-1 fold change PBS Treated Distant -5 0 5 10 15 20 25 PD-L1 fold change PBS Treated Distant 0 500 1000 1500 2000 2500 Tumor volume, mm3 66%* 57%* P=0.0002 P=0.004 P=0.01 P=0.16 P=0.003 P=0.02 Tilsotolimod Tilsotolimod Tilsotolimod Groups (n=11-12/group): 1. PBS 2. 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. PBS 2. 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 + Epacadostat 6. Tilsotolimod + Anti-PD-1 mAb 7. Epacadostat + Anti-PD-1 mAb 8. Tilsotolimod + Epacadostat + Anti-PD-1 mAb Days 0 Peritoneal tumor Tilsotolimod Epacadostat Anti-PD-1 mAb 15 16 17 18 19 8 9 10 11 12 Groups (n=11-12/group): 1. PBS 2. 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
Transcript of Triple combination of tilsotolimod (IMO-2125), epacadostat ......Tilsotolimod/aPD-1 PBS Tilsotolimod...
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.
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Treated tumor Distant tumor
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P=0.069
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P=0.206
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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
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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
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T i l s o t o l i m o d
Tilsotolimod
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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
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CT26.CL25
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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
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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
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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
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CT26.CL25
CT26.CL25
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Tilsotolimod i.t.
Epacadostat i.g.
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BACKGROUND
CONCLUSIONS
RESULTS
