Tumor Immunotherapy with BiTEs

Treating Cancer with T CellEngaging Antibodies Flavius Martin M.D. Vice President Research, Amgen Inc.

T cell Tumor Immunotherapy Players

Tumor Cell Antigens: - Surface/Presented BiTE® - Cell autonomous functions

T cells: - Response tuning - Effector function For Internal Use Only. Amgen Confidential.

2

BiTE®

Engineering an Adapter From Two Monoclonal Antibodies BiTE® = Bispecific T Cell Engager Tumor Cell-specific Antibody

BiTE®

T Cell-specific Antibody

Single-chain Antibody 1 VH

VH VL

VL Linker Single-chain Antibody 2

3

BiTE® Antibodies May Circumvent Frequent Escape Mechanisms

Any T Cell

CD3e

Do not Require T Cell Clone With Specific T Cell Receptor

Do not Require MHC Class I and Peptide Antigen for Recognition by T Cell

Can Make Any T Cell Recognize a Surface Antigen

TCR

X X

BiTE® Antibody

CD19 EpCAM CEA PSMA

Tumor Cell 4

A549 Lung Cancer Cells Peacefully Coexist With T Cells

Cancer Cell Resting CD8+ T Cell

A549 Lung Cancer Cell: T Cell Ratio = 1:1

5

Each T Cell Can Kill Nearby Cancer Cell After BiTE® AMG 110 Is Added

Apoptotic Cancer Cell

T Cell

Alive Cancer Cell

-EpCAM

-CD3

6

Kinetic Analysis of in-vitro BiTE® Reaction With Unstimulated T Cells AMG 330 = CD33/CD3-bispecific BiTE®; unstimulated human PBMC as effector cells at 10:1 ratio

T cell potency increases over time by 1-2 logs

28h

Complete lysis

22h 16h 10h AMG 330 [

7

Unstimulated T cells show lag phase before they start killing

Most Subpopulations of T Cells Can Contribute to Lysis by BiTE in vitro Kischel, R. et al., poster at AACR 2009 Relative Potency of Redirected Lysis

T Cell Populations Studied

Marker

CD8+ Bulk

CD8+

+++

CD8+ Naïve

CD28+ / CD45RA+

-/+

CD8+ EM

CCR7- / CD45RA-

++++

CD8+ EMRA

CD28- / CD45RA+

+++++

NKT Cells

CD3+ / CD56+

++

g/d T Cells

CD3+ /b -

+

CD4+ Bulk

CD4+

++

CD4+ Naïve

CD4+ / CD45RO-

-/+

CD4+ EM

CCR7- / CD45RA-

+++

CD4+ EM+CM

CD4+ / CD45RA-

++

CD4+ Naïve, Stimulated

CD69, CD25

+++

CD4+ EM, Stimulated

CD69, CD25

+++

8

Blinatumomab (AMG 103), a CD19/CD3bispecific BiTE® Antibody • Murine antibody construct of 55 kDa produced by CHO cell-based process • CD19 target is diagnostic marker for B cell malignancies – 100% of ALL cases are positive for CD19 – Expressed on leukemic stem cells with self renewal capacity – CD19 is activator of PI3 kinase

• Serum half life of AMG 103 is 1-2 hours  continuous i.v. infusion using port/minipump

• Multiple clinical trials in adult and pediatric ALL patients; NHL experience N

KD = 10-9 M

-CD19

KD = 10-7 M

-CD3 9

C

CD19-BiTE - Response and HSCT

n/N

95 % CI

Primary endpoint CR/CRh during the first two cycles

81 / 189

43%

36–50

Exploratory endpoints MRD response during the first two cycles CR/CRh

60 / 73

82%

72–90

CR, complete remission; CRh, complete remission with partial hematological recovery of peripheral blood counts; MRD, minimal residual disease (< 10-4)

Activity of Blinatumomab in NHL Patients Is Dosedependent (Phase 1 Study AMG 103-104) 12x higher dose needed for tumor

Peripheral T Cell Redistribution

Complete and Sustained B Cell Depletion Bone Marrow Clearance; First PR/CR High RR 0.5 5 1.5

15

30

60

Dose Levels Tested [µg/m2/Day]

11

90

Preliminary Learnings From Blinatumomab Monotherapy about BiTE® Modality • Need only very low doses – High response rates in ALL patients at steady state serum levels of 0.5 to 1 ng/ml ( 50,000/μl and ANC > 500/μl) cIV, continuous intravenous; HSCT, hematopoietic stem cell transplantation

Probability of Overall Survival

CD19-BiTE in ALL - Overall Survival 1.0

Overall Survival N=189

0.8

Median OS, months

0.6

95% CI, months

0.4

4.2–7.5

0.2 0 0

2

Patients at Risk 189 139

Probability of Overall Survival

6.1

4

6

8

10 Months

12

14

16

18

104

72

44

27

21

10

6

0

1.0

20

Overall Survival Landmark Analysis Day 77*

0.8

CR/CRh No CR/CRh N=79 N=50 Median OS, months 9.9 2.7 95% CI, months 6.8–NE 1.6–4.5

0.6 0.4 0.2 0 0

2

4

6

8

12

14

16

Patients at Risk 79 68 50 26

10 Months

43 15

33 9

21 5

12 4

8 2

5 1

0 0

*After two treatment cycles

NE, not estimable

18

20