Harpreet Singh, PhDCo-Founder & CSO

immatics biotechnologies GmbH, Tuebingen (Germany)

iSBTc 2007, Boston MA

IMA901- a novel multi-peptide vaccinefor treatment of renal cell carcinoma

2 TUMAPs = tumor-associated HLA-binding peptides

© Hans-Georg Rammensee (1991)

3 immatics biotechnologies

Approach

• Develop therapeutic cancer vaccines based on multiple peptides derived from tumor-associated antigens

• Use novel peptides confirmed to be naturally presented on primary tumor tissue

• Multi-peptide vaccines are fully synthetic and provided as stable, lyophilized formulation

• Perform multi-centre clinical trials with centralized and highly standardized immunomonitoring

4

1 2 3 4 5R1

R30

5

10

15

20

XPRESIDENT™ platform for identification of novel and naturally presented tumor-associated pepties

blood

Tumor cellsNormal cells

mRNA

Over-expression tumor vs. healthy

Gene Chips

HLA peptides

TUMAPsequences

LC-MS/MS

T cells

Selection of strongly immunogenic TUMAPs from

relevant tumor antigens

Synthesis of candidate TUMAPs

Collection of tissue samples

Correlation

ELISpot HLA tetramers

Immunoassays

A novel HLA-A*02-biding tumor-associated peptide fromc-met proto-oncogene

5

0.00.00.10.00.22.1

0.10.90.60.71.01.72.8

1.1 0.50.20.20.90.10.30.40.01.00.80.2

7.5

13.0

9.8

12.113.9

13.0

26.0

6.5

4.3

8.0

16.0

22.6

5.3

13.0

16.0

0.7

9.88.6

24.3

leukocytes

CD

3+

cells

CD

19

+cells

den

dritic cells

adren

al glan

d

blad

der

bon

e marrow

wh

ole brain

colon

heart

kidn

ey

liver

lun

g

placen

ta

prostate

salivary glan

d

small in

testine

skeletal mu

scle

spleen

stomach

testis

thym

us

thyroid

trachea

uteru

s

RC

C0

04

4T

RC

C0

06

8T_

a

RC

C0

07

0T

RC

C0

07

3T

RC

C0

07

5T

RC

C0

09

8T

RC

C0

10

3T

RC

C0

10

8T

RC

C0

11

2T

RC

C0

11

5T

RC

C0

11

6T

RC

C0

06

8T_

b

RC

C0

10

0T

RC

C0

11

9T

RC

C0

13

0T

RC

C0

13

3T

RC

C0

13

5T

RC

C0

14

6T

RC

C0

14

9T

rela

tive

expr

essi

on

203510_at

Healthy tissues RCC tumors

% S

peci

fi c L

ysi s

E:T

Croft (A2+Met-)

HCT 116 (colon)

K 562 (NK-sensitive)

SK-OV-3 (A2-Met+)

MZ 1257 (renal)MEL 1479 (melan.)SK-OV-3 (A2-Met+)

% S

peci

fi c L

ysi s

-10

0

10

20

30

40

0,1 1 10 100

-100

102030405060

0,1 1 10 100

E:T

A 498 (renal)

SK-OV-3 (A2-Met+)

K 562 (NK-sensitive)

Croft (A2+Met-)

MCF-7 (breast)

U 266 (myelom.)

K 562 (NK-sensitive)

SK-OV-3 (A2-Met+)

-20

0

20

40

60

80

0,1 1 10 100

0

10

20

30

40

50

0,1 1 10 100

6 TUMAP Development Platform

1. Shotgun Identification *

~100 promisingTUMAPs

7. Pharmaceutical development

2000-5000 peptides associated with selected type of cancer

Pre-clinical Development

Month 24 Month 12

Phase 1

Month 0

Product candidate comprising 10-12 TUMAPs

2. Expression analysis *3. Functional analysis4. TUMAP re-detection **5. TUMAP quantification **6. TUMAP validation *

8. GMP manufacturing9. Filing of IMPD/IND

~20 high-potentialTUMAPs

* patented / ** proprietary know how

Screen – Hit – Lead – Drug Candidate

7 IMA901

IMA901: renal cell cancer• status: phase 2 started in September 2007

• >200,000 new incidences worldwide (approx. 3% of all cancers),thereof approx. 60% late-stage

• 5-year survival rate <10% in stage IV disease

• Approved therapies: cytokines, TKIs (sorafenib, sunitinib, temsirolimus)

• RCC known as immunogenic tumor

• HLA class I and class II expression directly by tumor cells

HLA class II expression in healthy renal tisusue (endothelial cells and monocytes)

HLA class II expression in RCC

Histology by Karin Klingel, Tübingen

8Composition of IMA901 for treatment of renal cell carcinoma

# Peptide ID Allele Antigen Common acronyms and synonyms

1 IMA-ADF-001 HLA-A*02 Adipophilin adipose differentiation-related protein, ADRP

2 IMA-APO-001 HLA-A*02 Apolipoprotein L1 APOL1

3 IMA-CCN-001 HLA-A*02 Cyclin D1 CCND1, PRAD1, parathyroid adenomatosis 1, BCL-1

4 IMA-GUC-001 HLA-A*02 GUCY1A3 guanylate cyclase 1-soluble-alpha 3

5 IMA-K67-001 HLA-A*02 KIAA0367 --

6 IMA-MET-001 HLA-A*02 c-met proto-oncogene MET, HGF (hepatocyte growth factor) receptor, HGFR

7 IMA-MUC-001 HLA-A*02 MUC1 mucin, CD227, episialin, epithelial membrane antigen

8 IMA-RGS-001 HLA-A*02 RGS-5 regulator of G-protein signalling 5

9 IMA-ADF-002 HLA-A*02 Adipophilin adipose differentiation-related protein, ADRP

10 IMA-MMP-001 HLA-DR MMP7 matrix metalloproteinase 7

11 IMA-HBV-001 HLA-A*02 HBV core Antigen HBc, HBcAg, cAg

9

An open label study to evaluate safety and immunogenicity

of the peptide based therapeutic cancer vaccine IMA901injected intradermally with GM-CSF as adjuvant

in patients with renal cell carcinoma

Phase 1

Study Code IMA901-101

A. Stenzl, University Hospital of TübingenM. Stähler / Ch. Stief, Klinikum Grosshadern, Munich

P.Y. Dietrich, University Hospital of GenevaA. Haferkamp / M. Hohenfellner, University of Heidelberg

J. Beck, University of MainzT. Eisen, Royal Marsden Hospital, London

IMA901 Phase 1 Study

10 IMA901 Phase 1 Study

IMA901 phase 1 study outline• Design multi-centre, single arm phase 1

• Patients 28 patients with advanced renal cell carcinoma (HLA-A*02-positive)

• Scope 6 centers, 3 countries (DE, CH, UK)

• Dose 4.5 mg (400 µg per peptide) IMA901 i.d. 8x75 µg GM-CSF i.d. 8x

• Primary Endpoint Systemic safety, local tolerability

• Secondary Endpoints - Immunogenicity of IMA901- Pharmacokinetics intradermal GM-CSF- Any evidence of anti-tumor response

11IMA901 phase 1 Vaccination schedule

VACCINATIONGM-CSF _______________IMA901 ________________

WEEKDAY 85 - 92

Follow-up Period

VISIT 1.2.3 4 5 6 7 8

1 2 3 4 6 10–1–2 131.2.3 8 15 22 36 64–3–14

Screening Vaccination (Treatment) Period

Scr FU

T-CELLSAMPLE

FUV1 V4 V5 V6 V7 V8S2

Immunomonitoring: - peptide-specific T-cell responses (ELISpot/tetramer)- Foxp3+ Tregs pre and post vaccination

Tumor assessment: - according to RECIST criteria at screening and follow-up

12IMA901 Phase 1 ImmunomonitoringExample for raw data in tetramer assay

20060517a.039

FL2-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

0.01%99.98%

0.00%0.01%

20060517a.039

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060517a.039

FL1-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

20060517a.040

FL2-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

0.09%99.57%

0.00%0.34%

20060517a.040

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060517a.040

FL1-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

20060502a.044

FL2-H

010

110

210

310

410

FL4-

H0

10

110

210

310

410 0.04% 0.00%

99.96% 0.01%

20060502a.044

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060502a.044

FL1-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

20060503a.029

FL2-H

010

110

210

310

410

FL4-

H

010

110

210

310

410 0.01% 0.00%

99.55% 0.44%

20060503a.029

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060503a.029

FL1-H

010

110

210

310

410

FL4-

H0

10

110

210

310

410

05-001 S2+V1FL1: CD8FL2: rK67-001FL4: rCCN-001

05-001 V4+V5FL1: CD8FL2: rK67-001FL4: rCCN-001

03-003 V4+V5FL1: CD8FL2: rK67-001FL4: MET-001

03-008 V6+V7FL1: CD8FL2: MUC-001FL4: ADF-002

13T-cell Responses in Phase 1IMA901-101 Immunomonitoring

Vaccine-induced T-cell responses

• N=27 patients evaluable for immune response• T-cell response measured with ELISpot and tetramer assays

0%

52%

85%

74%

30%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

Negative controlpeptide (HIV)

Viral marker peptide(HBV)

At least 1 response toany TUMAP or HBV

At least 1 response toany TUMAP

Multi-TUMAPresponses

Vacc

ine-

indu

ced

imm

une

resp

onde

rs

14IMA901 Phase 1 ImmunomonitoringKinetics

T-cell response kinetics (representative patient)

Patient 03004

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

S2 V1 V4 V5 V6 V7 V8 FU

Scheduled timepoint (scale in weeks)

% T

etra

mer

+ am

ong

CD

8+ T

-cel

ls

TUMAP PoolHBV-001HIV-001

15IMA901 Phase 1 ImmunomonitoringEx vivo phenotyping

Phenotyping of T-cell response (ex vivo, N=1)

• Pre-vaccine T cells are of naïve phenotype

• Post-vaccine T cells are of effector memory phenotype

20060727a.021

FL3-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

20060727a.021

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060727a.022

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060727a.022

FL3-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

20060727a.023

FL1-H

010

110

210

310

410

FL2-

H

010

110

210

310

410

20060727a.023

FL3-H

010

110

210

310

410

FL4-

H

010

110

210

310

410

01-009 V1FL1: CD45RAFL2: CCR7FL3: CD8FL4: rCCN-001

01-009 V5FL1: CD45RAFL2: CCR7FL3: CD8FL4: rCCN-001

01-009 V7FL1: CD45RAFL2: CCR7FL3: CD8FL4: rCCN-001

16IMA901 Phase 1 ImmunomonitoringExample for Treg quantification

20060725a.044

FSC-H0 256 512 768 1024

SS

C-H

0

256

512

768

1024

lymphocytes

20060725a.044

FL1-H

010 110 210 310 410FL

3-H

010

110

210

310

410 CD45+ CD4+ CD45+

20060725a.044

FL1-H

010 110 210 310 410

FL2-

H

010

110

210

310

410

39,71%57,39%

2,72%0,18%

20060725a.050

FSC-H0 256 512 768 1024

SS

C-H

0

256

512

768

1024

lymphocytes

20060725a.050

FL1-H

010 110 210 310 410

FL3-

H

010

110

210

310

410 CD45+ CD4+ CD45+

20060725a.050

FL1-H

010 110 210 310 410FL

2-H

010

110

210

310

410

36,25%56,54%

7,02%0,18%

Gated on CD45+ lymphocytes -> automatic quadrant setting!

Patient 03-003pre-vacc.

Patient 03-006pre-vacc.

FL1: CD4FL2: Foxp3FL3: CD45

Gated on all cells. Gated on lymphocytes.

17IMA901 phase 1 immunomonitoringRegulatory T cells and T-cell response

Patients with multiple TUMAP responses have significantly lower TREGlevels in the periphery than patients with 0-1 TUMAP responses (p=0.016 Wilcoxon Test, N=26 pts)

0.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

-1 0 1 2 3 4

No. of TUMAP responses in patients

% F

oxp3

+ Tr

egs

of C

D45

+ ly

mph

ocyt

es

Pre-vaccinationPost-vaccinationAverage Pre-v.Average Post-v.

18Efficacy - Change of tumor size and T-cell response (ITT, n=28)

17 158

0024 2 0 0 0 0 00

52

145

25

46

10

263133

-27-38

-9-6-3

-60

-40

-20

0

20

40

60

80

100

120

140

16004

-001

05-0

05

03-0

12

03-0

13

01-0

07

05-0

02

01-0

01

01-0

10

05-0

04

03-0

04

03-0

06

03-0

02

01-0

11

03-0

15

03-0

05

03-0

09

03-0

16

01-0

02

01-0

03

01-0

08

02-0

01

03-0

01

05-0

01

03-0

03

06-0

04

01-0

09

03-0

08

03-0

14

patient numbers

% c

hang

e fr

om b

asel

ine

% increase % decrease

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

New

lesi

ons

MET

-001

MU

C-0

01, G

UC

-001

CC

N-0

01

ADF-

002

CC

N-0

01

CC

N-0

01, M

ET-

001

CC

N-0

01, K

67-0

01

CN

N-0

01

CC

N-0

01

CC

N-0

01

CC

N-0

01, A

DF-

001,

GU

C-0

01

CC

N-0

01, M

ET-

001,

GU

C-0

01

MU

C-0

01,

CC

N-0

01, A

DF-

001

CC

N-0

01, K

67-0

01

CC

N-0

01

AD

F-00

2, A

PO

-001

CC

N-0

01

MET

-001

not e

valu

able

ADF-

002

CC

N-0

01

No

FU tu

mor

ass

essm

ent

New

lesi

ons

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

HBV

resp

onde

r

19IMA901 Clinical Study Phase 1Conclusions

IMA901 is safe and well tolerated (data not shown)

IMA901 is immunogenic

• Vaccine-induced immune responses in 74% of pts.

• Multiple vaccine-induced responses in 30% of pts.

Multiple vaccine-induced immune responses to IMA901

• seem to inversely correlate with the level of regulatory T cells prior to vaccinaton (p=0.016)

• seem to correlate with the clinical outcome (partial response and stable disease according to RECIST) (p=0.015)

Next: multi-centre phase 2 trial in Europe (started Sept 2007)

• ~70 met RCC pts., 2nd line after TKI or cytokine therapy failure

• continous vaccination for 9 months, evaluation of the disease control rate at 6 months

• Evaluation of the impact of low-dose cyclophosphamide on Tregs, MDSC and immune responses in a randomized fashion (+/- CY)

20

Broad-spectrum tyrosine kinase inhibitors (TKIs) were recently approved for treatment of metastatic RCC pts

Question: can TKIs be combined with vaccination simultaneously or sequentially?

Assessment of impact of sorafenib and sunitinib on immune cells in vitro and in vivo

Impact of sunitinib and sorafenib on immune cells

21 Impact of sunitinib and sorafenib on immune cells

Sorafenib but not sunitinib inhibits human T-cell activation in (allogeneic) mixed lymphocyte culture

• Data not shown: very similar observation for CD4+ T cells

CD8+

0,0%

10,0%

20,0%

30,0%

40,0%

50,0%

60,0%

70,0%

A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+ A2+

none A2+ A2+ A2+ A2- A2- A2- A2- A2- A2- A2- A2- A2- A2-

DMSO none Sorafenib13 µM

Sunitinib250 nM

none DMSO Sorafenib1.6 µM

Sorafenib3.3 µM

Sorafenib6.5 µM

Sorafenib13 µM

Sunitinib31 nM

Sunitinib63 nM

Sunitinib125 nM

Sunitinib250 nM

Effector / Target / TKI

% C

FSE-

/ A

2+ C

D8+

Lym

phoc

ytes

22 Impact of sunitinib and sorafenib on immune cells

Sorafenib and sunitinib have no impact on human T-cell activation by artificial peptide-presenting APCs in vitro

Melan-A-specific CD8+ T-cell in vitro priming

HBC-133

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

Mock Mock Sunitinib250 nM

Sunitinib125 nM

Sunitinib63 nM

Sunitinib31 nM

Sorafenib13 µM

Sorafenib6.5 µM

Sorafenib3.3 µM

Sorafenib1.6 µM

DDX5-001HD

MLA-001HD

MLA-001HD

MLA-001HD

MLA-001HD

MLA-001HD

MLA-001HD

MLA-001HD

MLA-001HD

MLA-001HD

% M

LA-0

01+

23 Impact of sunitinib and sorafenib on immune cells

Sorafenib but not sunitinib inhibits the LPS-mediated maturation of human mDCs in vitro

• Data not shown: no effect of sunitinib on maturation of mDCs

• Data not shown: sorafenib but not sunitinib affects the migrations capacities of mDCs and downmodulates CCR7

Hipp et al., submitted

24 Impact of sunitinib and sorafenib on immune cells

Combination of peptide vaccination in mice and TKI simultaneously: sorafenib but not sunitinib inhibits OVA peptide-induced T-cell responses in C57BL/6 mice

Hipp et al., submitted

OVA-specific T cells (tetramer) CD4+CD25hi Tregs

25

Sunitinib is compatible with• in vitro antigen-induced T-cell expansion (human and

mouse)

• in vitro TLR-mediated DC maturation (human)

• in vivo peptide-induced T-cell proliferation (mouse)

On the other hand, sorafenib significantly inhibits all of these immunological endpoints• but: the inhibition by sorafenib is reversible within days in

mice (not shown)

Sunitinib but not sorafenib slightly decreases regulatory T cell levels in miceSorafenib but not sunitinib affects MyD88-dependent and MyD88-independent signaling pathways in APCs (not shown)

Impact of sunitinib and sorafenib on immune cellsConclusions

26

Peter BrossartMadeleine HippDaniela WerthKatharina Brauer

Arnulf StenzlMarkus KuczykSusan FeyerabendJörg Hennenlotter

Hans-Georg RammenseeStefan StevanovicCécile Gouttefangeas

Co-workers and CollaboratorsIMA901 Pre-clinical and Phase 1 / TKI project

IMA901 Discovery TeamToni WeinschenkClaudia LemmelJörn DengjelOliver SchoorNorbert HilfMartin PriemerSilvia RothBettina StreileSylvia ZwickerAntje ArthurColette Song

IMA901-101 Immuno-monitoring TeamSteffen WalterNicole ArnoldMonique GoerlVerona Vass

IMA901-101 CMC TeamPeter LewandrowskiWerner StüberJessica BergmannChristian FlohrVerena Beyer

IMA901-101 Clinical TeamJürgen FrischAndrea MayerAnnette SchmidTamara SpürginAngela Arena

Scientific Advisory BoardHans-Georg RammenseeCornelius MeliefChristoph HuberPedro RomeroCraig Slingluff Jr.