Download - Poster ImprimePGG (Imprime) plus pembrolizumab(PEM): a ...€¦ · 4. Innate effector activation • Promotes tumor killing – Macrophages, neutrophils • Alleviates immunosuppression

Abstract

Imprime PGG(Imprime)pluspembrolizumab (PEM):aphase2immunotherapeuticcombinationinpatientsselectedforanImprime-specificbiomarker.

StevenO’Day1, Nandita Bose2,MarkUhlik2,Radha Prathikanti2,BenHarrison2,StevenLeonardo2,RichardHuhn2, NadineOttoson2,Xiaohong Qiu2,RichardWalsh3,PauletteMattson2,MableMa2,KatieErtelt2,JamieLowe2,MicheleGargano2,MichaelChisamore5,BrunoOsterwalder4,JeremyRGraff21JohnWayneCancerInstitute,SantaMonica,CA90404,2BiotheraPharmaceuticals,Inc.Eagan,MN55121,3ImmunoResearch,Eagan,MN55121,4B.O.ConsultingGmbH,Riehen,SwitzerlandCH-41255Merck&Co.,Inc.,Kenilworth,NJ,USA

Poster#733LB-A31

AACR-NCI-EORTCMolecularTargetsandCancerTherapeutics,BiotheraPharmaceuticals,Inc.,Philadelphia,PA,Oct26-30,2017

Imprime PGG (Imprime) is a novel immunotherapeutic that acts as a non-self danger signal to activatethe innate immune system and coordinate an adaptive immune system response. In multiple preclinicaltumor models, Imprime significantly enhances anti-tumor efficacy of multiple immune checkpointinhibitors (CPI). Mechanistically, Imprime directly binds to innate immune effector cells via theformation of an immune complex with naturally-occurring anti-b glucan antibodies (ABA). This immunecomplex is required for Imprime to activate innate effector functions- monocyte and dendritic cellactivation, and enhanced cytokine production. In a recently completed healthy volunteer study,activation of innate immune cell functions was demonstrated after intravenous (IV) infusion of Imprimein subjects with pre-treatment [Tx] ABA levels > 20 µg/ml. In ex vivo human donor blood studies, wholeblood from subjects with low (i.e. insufficient) ABA failed to show innate immune activation unlessrescued by the addition of purified ABA (ABA from the serum of high ABA donors) or commercialimmunoglobulin (IVIg). Retrospective analyses of data from previous clinical trials showed that higherpre-Tx ABA levels correlated with enhanced overall survival in patients (pts) treated with Imprime.Collectively, these data support the hypothesis that ABA are essential to the therapeutic benefit ofImprime. A phase 2 clinical trial combining Imprime (4 mg/kg weekly IV) with Pembro (200 mg, q3w)has begun in metastatic melanoma pts who have failed a CPI and in CPI naïve, triple-negative breastcancer (TNBC) pts who have failed chemotherapy. Pts are screened for sufficient ABA (> 20 µg/ml) byELISA. Pre- and early-on Tx biopsies (6 wks) and at time of response/progression or end of Tx arerequested to assess immune cells at the tumor site by multispectral immunofluorescence. Bloodsamples are collected pre- and post- Imprime infusion on Day 1 of each treatment cycle for the first 6cycles and q4 cycle thereafter. The first TNBC pt had a pre-Tx ABA of 31 µg/ml. Following 6 wks of Tx,the pt’s total target lesion mass was reduced >40%. Partial response continued at 12, 18 and 24 wks(>50% reduction vs baseline). A biopsy of a remaining pelvic lesion after 18 wks Tx revealed primarilyfibrotic tissue. Blood samples showed evidence of innate immune activation including increasedexpression of MCP-1, IL-8, chemokines targeting CCR5 and CXCR3 that are associated with enhanced Tcell infiltration and effector function. The first melanoma pt (pre-Tx ABA 42 µg/ml) showed a modestincrease in lesion size at 12 wks (+12% versus baseline). Pre- and on-Tx (6 wk) biopsy material (from twodifferent lesion locations) was obtained for multichannel immunofluorescence. The pre-Tx biopsy tumorsample was primarily devoid of immune cells. In contrast, the on-Tx biopsy tumor sample indicatesextensive infiltration of the tumor bed by CD8 and CD4 T cells, and innate immune cells. PreviousImprime clinical and ex vivo data suggest that sufficient ABA may be important for Imprime-mediatedinnate immune activation. Early clinical observations in ABA pre-selected patients support the notionthat Imprime may provide benefit in combination with Pembro in these two populations.

Imprime MechanismofAction

Cytokine Chemokine Production

IFNs

MonocyteMacrophageDendritic CellNeutrophil

Proposed mechanism of action ignites a fully functional immune response against cancer

1. Imprime forms an immune complex• Anti-β glucan antibodies (ABA) (IgG2a)• Complement fragment (iC3b)• Imprime-β glucan

2. Binds to and co-ligates 3 surface receptors• ABAs bind Fcγ receptor IIa (CD32a)• iC3b binds complement receptor 3 (CR3)• Imprime binds dectin-1 receptor

3. Receptor co-ligation provides activation signal• Type 1 interferon gene expression• Chemokines/cytokines

4. Innate effector activation• Promotes tumor killing– Macrophages, neutrophils

• Alleviates immunosuppression– M2èM1 repolarization– MDSC maturation/differentiation

• Activates antigen presentation– Dendritic cell maturation– M1 APCs

Imprime PGG: An Immunological “Ignition Switch”

IgG ABA Complement opsoniniC3b

iC3b

4

Imprime andCPI:PreclinicalEfficacy

Imprime

ABA (RAU/mL): 0 350 700

0500

100015002000

15000

20000

25000

IL-8MCP-1

Che

mok

ine

(pg/

mL)

0 10 20 30 40 50 60 700

20

40

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80

100

Citrate + RajiImprime + RajiImprime + 175 RAU/mL ABAImprime + 250 RAU/mL ABAImprime + 350 RAU/mL ABA

Time (min)

Reac

tive

Oxyg

en S

pecie

s (R

OS) i

n RL

U

PD Effects ROS Generation

Left Panel- Imprime induced pharmacodynamic effects (MCP-1, IL-8) are enhanced in whole blood from a “low binder” by purified ABA supplementation. Right Panel- The generation of Reactive Oxygen Species (ROS) is enhanced in neutrophils isolated from a “low binder” when exposed to rituximab-opsonized Raji B cell lymphomas. Note: Dose dependent ROS generation with ABA supplementation. RAU = relative antibody units.

ABA Supplementation in a Non-Responsive Donor Rescues ImmunoPD Effects and Functional Tumor Cell Killing Activities

Neutrophils

Monocytes

Vehicle Imprime PGG Imprime PGG + ABA

Imprime BindingCanBeRescuedbyABASupplementationin“LowBinding”Individuals

Wholebloodwasisolatedfromalow-bindinghealthyhumandonor.Imprime wasdetectedonthesurfaceofNeutrophilsandMonocytesusingananti-βglucan imagingantibodyBfDIV followedbyflowcytometry.BindingwassubstantiallyenhancedbytheadditionofpurifiedhumanIgG ABAasindicatedbytheshiftup(rightpanels).

Imprime andABA:ExVivoStudies

Imprime andABA:RetrospectiveStudies

HumanHealthyVolunteerStudy

CONFIDENTIAL

Imprime PGG Enhances the Response to Checkpoint Inhibitor Therapy

0

10

20

30

40

50

60

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100

Control Imprime PGG only

PDL-1 only Imprime PGG + PDL-1

% of

Inje

cted

mice

with

out t

umor

(d

ay 29

)

5.6% 11%

33%

83%*

Control Imprime PGG αPD-L1 Imprime + αPD-L1MC-38 colon cancers were subcutaneously injected into C57BL/6 mice. 3 days later, treatment was initiated. N = 18 per group except Imprime PGG + PD-L1, n =17. Tumor-free mice were re-injected in the opposite flank with MC-38 and remained tumor-free for another month, without any additional therapy. Tumors grew in all age-matched, tumor naïve control mice injected with MC-38. * ANOVA analysis, Tukey adjustment, p = 0.001 vs. aPD-L1, p < 0.0001 vs vehicle control. Differences between all other groups were not significant ( p ≤ 0.05).

Tumor-free mice were re-challenged with MC-38 cells on the opposite flank and remained tumor-free. This suggests the

establishment of immunological memory.

1.0

0.8

0.2

0.0

0.6

0.4

0 10 20 30 40 50

ABA ≥ 35

ABA < 35

HR=0.40 (0.22-0.73)p=0.0018

Surv

ival P

roba

bilit

y

85 36 8 1 0 027 17 6 2 1 1

Significant (p<0.05) tests: 36 out of 86 (41.9%)

Imprime + Cetuximab

2

1

0.5

10 20 30 40 50

Imprime + Cetuximab

1.0

0.8

0.2

0.0

0.6

0.4

Surv

ival P

roba

bilit

y

HR=0.24 (0.1-0.55)p=0.00031

ABA > 45

ABA < 45

0 10 20 30 40 50

0 10 20 30 40 50

ABA < 20

ABA ≥ 20

1.0

0.8

0.2

0.0

0.6

0.4

HR=0.77 (0.49-1.22)p=0.27

Surv

ival P

roba

bilit

y

% Subjects- ABA levels:49% ≥2024% ≥35 16% ≥45

57 25 5 1 0 55 28 9 2 1 1

94 40 10 1 0 18 13 4 2 1 1

ABA IgG (μg/ml)

CRC Primus Trial: Higher Pre-Treatment ABA Levels Correspond with Improved Overall Survival

0.25

0.13HR w

ith 9

5% C

I

Retrospective analyses: Primus trial, 3rd line CRC cetuximab ± Imprime. Pre-treatment ABA levels were measured by ELISA. Sample “cutpoints” for ABA are noted at 20, 35 and 45 !g/ml.

0 20 40 60 80 100 120 140 160 180 200 220

ABA IgG (ug/ml)

Don

ors

Distribution of ABA IgG in Lung Cancer Samples

53.5%

0 20 40 60 80 100 120 140 160 180 200 220

ABA IgG (ug/ml)

Dono

rs

Distribution of ABA IgG in Breast Cancer Samples

45.8%

0 20 40 60 80 100 120

ABA IgG (ug/ml)

Dono

rs

Distribution of ABA IgG in Ovarian Cancer Samples

58.6%

IgG ABA Distribution in Patients with Different Cancers

Biothera Trials

Indication n median IgG ABAColon (Primus) 169 18.7 μg/mlNSCLC (0821) 58 20.4 μg/mlNSCLC (0822) 59 23.0 μg/ml

Tumor(EphA2+Ki67+)

CD8 T-cells(CD3+CD8+)

Activated CD8 T-cells(CD3+CD8+Ki67+GrzBHigh)

Vehic

leIm

prim

e

Increased Activation of T-Cells in the MC-38 Tumor Bed after Imprime Treatment

MC-38- Imprime dosed IV 1.2mg/mouse twice weekly. 10 days treatment. Tumor Tissues imaged using the Perkin Elmer Vectra Multi-spectral Imaging System.

Phase2Imprime +Pembro Studies Imprime andPembro:CPIExperiencedMetastaticMelanoma

Pre Treatment Sample Post Treatment Sample

2mm 800 µm


Patient 103103

Hematoxylin and Eosin Stains for Pre- and On-Tx Melanoma Biopsies

Pharmacodynamic Effects of Imprime are Restricted to Biomarker Positive Human Subjects

Blue=ABAIgG+Red =ABAIgG-/IgM-Black=ABA IgG-/IgM+

0.10

1.00

10.00

100.00

1000.00IL-6

IL-8

IP-10

MCP-1

MIG

MIP-1 ALPHA

MIP-1 BETA

TNF-ALPHA

Cytokine/chemokine expression. At the end of infusion, serum was taken to assesscytokine and chemokine expression changes versus pre-dose values in each subject (foldincrease shown) using the Luminex XMAP technology.

NSCLCMetastatic Melanoma

Triple-Negative Breast Cancer

Head and Neck Cancer

Phase Phase 1b/2 Phase 2 Phase 2 Phase 2

Combination Therapy Pembrolizumab Pembrolizumab Pembrolizumab Pembrolizumab

Patients 2nd line patients 2nd line patients- Pembro failure

2nd- 3rd line patients

- Pembro Naive

• Stable disease >3 mosPembro

• Pembro Failure

Number of Subjects 36 29 42 87

Randomization Single arm Single arm Single arm Single arm

Primary Endpoint PFS ORR ORR ORR

Secondary Endpoints Safety, PFS, OS Safety, PFS, OS Safety, PFS, OS Safety, PFS, OS

Initiation 3Q 2016 4Q 2016 4Q 2016 1Q 2017

Phase 2 Clinical Studies: Imprime + Pembrolizumab

24

Patients selected for Imprime-specific biomarker (Anti-Beta glucan Antibodies, ABA > than 20 μg/ml)


DAPITumorCD4CD8GrzB

Fold Increase vs Pre-TxCytotoxic T cells (CD8)

Total number = 28XProliferating = 33XActivated = 50X

Helper T Cells (CD4)

Total Number= 58XProliferating = 73XActivated = 24X

Lymphocyte Infiltration: Pre and On Tx Biopsy


NucleiTumor

Mac/MonoCD80PD-L1

Myeloid Character Post vs Pre TxMyeloid Infiltration 56X increaseCD80+ 100X increasePD-L1+ Myeloid 13X increase

Increased Myeloid Infiltration and Activation Post Treatment

Pre-Treatment Post-Treatment

% of Myeloid Cells CD80+: 6.34%% of Myeloid Cells PD-L1+: 0.90%% of Myeloid Cells CD206+: 18.45%% CD80+/CD206+ (M1:M2 ratio): 0.34

TumorCD163CD206CD80

% of Myeloid Cells CD80+: 13.79%% of Myeloid Cells PD-L1+: 14.88%% of Myeloid Cells CD206+: 3.11%% CD80+/CD206+ (M1:M2 ratio): 4.44

Evidence for increased M1:M2 Character Post TreatmentTranslational Research: Schedule of Assessments

Cycle Screening 1 2 3 4 5 6 10,Q4C EndofTrtmnt

Week Tubetype Upto-4 1 4 7 10 13 16 28

CBC/Diff:Preand4hrPostSOI ETDA X X X X X

PBMC:PreandPost Heparin X X X X

PAXGene (RNA):Pre-ImprimeandPostPembro PAXGene X X X X X

ABA:PreandPostImprime Serum X X X X X X X X X

Cytokines:PreandPostImprime Serum X X X X X X X X

ImprimePK:Pre,1.5hr postEOIImprime Serum X X

Pembo PK:PrePembro and3hrpostEOIPembro Serum X X

Biopsies(archival,pre-C1,pre-C3andupontimeofresponse/progression Tissue X X X

SOI:StartofInfusion

Imprime- Induced PD Effects Are Restricted to ABA + Subjects

0

20

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100

Circu

lating

Immu

ne Co

mplex

es (E

q ug/m

l)

Longitudinal CIC

253967

10345570

Imprime Dose Week #1

Subjects

0

250

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1000

1250

1500

Mono

cytes

(cell

/ul)

Monocyte Mobilization

25

67

10345570


Subjects

39

0

2500

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10000

12500

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20000

Neutr

ophil

s (ce

ll/ul)

Neutrophil Mobilization

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67

10345570


Subjects

39

0

2500

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10000

12500

15000

SC5b

-9 (ng

/ml)

Complement

25

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103455


Subjects

39

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10000

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30000

MCP-1

(pg/m

l)

Cytokines

25

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103455


Subjects

39

70

Biomarker + (ABA ≥ 20μg/ml)Biomarker – (ABA < 20μg/ml)

Immune Complex Monocyte Mobilization Neutrophil Mobilization

Complement Cytokines

Whole blood or serum was drawn from healthy volunteers at various time points before and after a single dose of Imprimeinfusion. ABA were measured in serum by ELISA. Circulating Immune complex formation was measured using theMicroVue Complement CIC-Raji Cell Replacement Kit. Cell mobilization was measured by complete blood cell counts, plusdifferentials. Monocyte and Neutrophil numbers are shown. Complement activity was measured by ELISA using the SC5b-9 Plus kits (Quidel). Cytokines and chemokines were measured in serum using Luminex XMAP technology. Fold over pre-dose values are plotted. Data from subjects who were treated with Imprime (4 mg/kg) and no pre-medications.

CONFIDENTIAL

Cycle 1 Wk 1-pre

Cycle 1 Wk 1-EOI

Cycle 2 Wk 4-pre

Cycle 2 Wk 4-EOI

Cycle 3 Wk 7-pre

Cycle 3 Wk 7-EOI

Cycle 4 Wk 10-pre

Cycle 4 Wk 10-EOI

Cycle 5 Wk 13-pre

Cycle 5 Wk 13-EOI

012345

10

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fold

ove

r pre

-C1

SC5b9 Fold changes 103102

fold over Pre-C1

Cycle

1 Pre-

Infusion

Cycle

1 EOI

Cycle

2 Pre-

Infusion

Cycle

2 EOI

Cycle

3 Pre-

Infusion

Cycle

3 EOI

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Infusion

Cycle

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Cycle

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Infusion

Cycle

5 EOI

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Patient 103102CIC vs ABA

CIC

CIC

ABAABA IgG (µg/m

l)

Cycle 1 Pre-Infusion

Cycle 1 EOI


Cycle 2 EOI


Cycle 3 EOI


Cycle 4 EOI


Cycle 5 EOI0

20

40

60

80

0

100

200

300

Patient 103102CIC vs ABA

CIC

CIC

ABAABA IgG (µg/ml)

Cycle

1 Wk 1

-pre

Cycle

1 Wk 1

-EOI

Cycle

2 Wk 4

-pre

Cycle

2 Wk 4

-EOI

Cycle

3 Wk 7

-pre

Cycle

3 Wk 7

-EOI

Cycle

4 Wk 1

0-pre

Cycle

4 Wk 1

0-EOI

Cycle

5 Wk 1

3-pre

Cycle

5 Wk 1

3-EOI

012345

10

20

30

40

fold

ove

r pre

-C1

SC5b9 Fold changes 103102

fold over Pre-C1

Circulating Immune complex formation was measured using a commercial immuno assay system (MicroVue Complement CIC-Raji Cell Replacement Kit). Complement activity wasmeasured by ELISA using the SC5b-9 Plus kits (Quidel).

TNBC 103102: ABA, Immune Complex Formation and Complement Activation TNBC Pt 103102: Induction of Imprime-Responsive Cytokines

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

5

10

15500600700800900

1000

pg/m

l

IL-8

IL-8

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

020406080

100

1000200030004000

pg/m

l

MIP-1 beta

MIP-1 beta

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

010203040

100200300

100015002000

pg/m

l

RANTES

RANTES

LOD1: 0.78LOD2: 0.99

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

50

100

150

pg/m

l

Eotaxin

Eotaxin

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

020406080

2000

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6000

pg/m

l

MCP-1

MCP-1

LOD1: 1.23LOD2: 2.43

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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pg/m

l

MIP-1 alpha

MIP-1 alpha

LOD1: 0.49LOD2: 1.32

Cytokines and chemokines were measured in serum using Luminex XMAP technology.

TNBC Pt 103102: Induction of Imprime-Responsive Cytokines

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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pg/m

l

GM-CSF

GM-CSF

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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10

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pg/m

l

GRO-alpha

GRO-alpha

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

05

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2000

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pg/m

l

IL-6

IL-6

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

1

2

3

4

5

pg/m

l

IL-7

IL-7

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

050

100150200

50000

100000

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pg/m

l

IL-1RA

IL-1RA

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

050

100150200

500

1000

1500

pg/m

l

IP-10 (CXCL10)

IP-10 (CXCL10)

LOD1: 0.51LOD2: 1.78


Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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pg/m

l

IL-18

IL-18

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

1

2

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4

pg/m

l

IL-15

IL-15

Cycle 1 Week 1 PRE

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Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

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OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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100

150

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pg/m

l

ITAC (CXCL11)

ITAC (CXCL11)

LOD1: 4.13LOD2: 6.4

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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1000

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pg/m

l

SDF-1 alpha

SDF-1 alpha

LOD1: 2.6LOD2: 18.11

Cycle 1 Week 1 PRE

Cycle 1 Week 1 EOI

Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

Cycle 4 Week 10 E

OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

0

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10

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pg/m

l

IL-2

IL-2

Cycle 1 Week 1 PRE

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Cycle 2 Week 4 PRE

Cycle 2 Week 4 EOI

Cycle 3 Week 7 PRE

Cycle 3 Week 7 EOI

Cycle 4 Week 10 PRE

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OI

Cycle 5 Week 13 PRE

Cycle 5 Week 13 EOI

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pg/m

l

IL-22

IL-22

TNBC Pt 103102: Cytokine Induction


TNBC Patient Melanoma Patient

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IL-7Eotaxin

GRO-alpha

IL-8

IP-10(CXCL10)

ITAC (CXCL11)

MCP-1

MIP-1alpha

MIP-1beta

RANTES

SDF-1alpha

IFN-alpha

IFN-gamma

IL-1alpha

IL-1betaIL-2IL-6TNF-alphaIL-12p70

IL-1RA

IL-10

IL-27

TNF-beta

IL-15

IL-18

IL-21

IL-17A

IL-22

IL-23

IL-4

IL-5

IL-9IL-13

IL-31

Chemokines

0.1

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1000GM-CSF

IL-7Eotaxin

GRO-alpha

IL-8

IP-10(CXCL10)

ITAC (CXCL11)

MCP-1

MIP-1alpha

MIP-1beta

RANTES

SDF-1alpha

IFN-alpha

IFN-gamma

IL-1alphaIL-1beta

IL-2IL-6TNF-alphaIL-12p70IL-1RA

IL-10

IL-27

TNF-beta

IL-15

IL-18

IL-21

IL-17A

IL-22

IL-23

IL-4

IL-5

IL-9IL-13

IL-31

C1 C2 C3InflammatoryCytokines

Th2

Th17

Cytokine Profiles: TNBC Patient and Melanoma Patient

Cytokines and chemokines were measured in serum using Luminex XMAP technology. Eachring radiating from the center represents an order of magnitude increase in expression.

Imprime andPembro:CPINaïveTNBC Imprime +Pembro:EarlyExperience

CONFIDENTIAL

Phase 2 Imprime + Pembrolizumab: Early Clinical Experience

Triple Negative Breast Cancer Patient 103102Objective ResponderTumor Shrinkage- 44% evident at week 6Tumor Shrinkage > 50% for > 28 weeksTumor Biopsy at ~ week 18- no tumor, fibrotic tissueIncreased ABA after Imprime dosingEvidence for Imprime-induced cytokines/ chemokines

Metastatic Melanoma Patient 103103Tumor Biopsy at week 6 suggests activated T cell InfiltrationTumor Biopsy at week 6 shows myeloid infiltration and activationEvidence for Imprime- induced cytokines/ chemokinesPatient off therapy at week 18

Early clinical experience provides evidence of clinical response (TNBC) and 1st proof of mechanism- i.e. that Imprime may drive immune activation at the tumor bed