Designing clinical trials with BiTE antibody constructs by leveraging from nonclinical data. Benno Rattel Biologics Congress Berlin, 2015

Transcription

1 Designing clinical trials with BiTE antibody constructs by leveraging from nonclinical data Benno Rattel Biologics Congress Berlin, 215 1

2 BiTE Antibody Contructs are Designed to Function as a Bridge between T Cells and Cancer Cells 2

3 Key Hallmarks of BiTE Mode of Action (1) Strictly target cell-dependent activation of resting T cells by BiTE (2) Highly potent and complete lysis of target cell by BiTE activated T cells (3) Lysis of dividing as well as non-dividing target cells (4) Serial lysis by BiTE -activated T cells (5) Sustained proliferation of BiTE -activated T cells (6) Does not require MHC Class I and peptide antigen for recognition by T cell (7) Does not require T cell clone with specific T cell receptor Hoffmann, P. et al. Intl J Cancer 25; 115:

4 CD19-CD3 bispecific BiTE antibody construct Clinically developed for treatment of r/r ALL Received breakthrough designation from the US FDA in July 214 BLA submitted to FDA in Sept 214 and to EMA in Oct 214 4

5 FDA Approved BLINCYTO TM on December 3rd, 214 5

6 The New BiTE Platform is NHP Cross reactive BLINCYTO TM New Platforms v3. Target Human and Chimpanzee Human and NHP T Cells (CD3) Human and Chimpanzee Human and NHP 6

7 Pharmacological Characterization

8 In Vitro Pharmacology Package for First in Human (FiH) Study Side-by-side comparison of pharmacology in human and toxicology species systems For bispecific binding binding affinity for human and Cyno target antigens epitope mapping specificity of binding 8

9 In Vitro Pharmacology Package for FiH Study Continued For redirected lysis dose-response of target cell lysis with various target cells correlation of EC 5 of lysis with target expression level specificity of target cell lysis kinetics of lysis dependence on effector to target ratios target cell lysis by various T cell populations PBMC / T cell donor variability redirected lysis of patient-derived malignant cells by autologous T cells (ex vivo PoC) 9

10 In Vitro: Blinatumomab Redirected Lysis A 1 GRANTA-519 B 1 EHEB C HBL D MEC-1 Redirected Lysis (EC 9 ) of Human B Lymphoma Lines at approx..5 ng/ml Specific Lysis [%] E NCEB F Karpas MT13 [ng/ml] Amgen Report No. 13-PCD-61 1

11 Estimation of a Therapeutic Dose Based on In Vitro PD Data EC 9 of BLINCYTO TM -mediated redirected lysis: ~.5 ng/ml In the clinic, anti-tumor activity was observed at a c.i.v dose of 15 µg/m 2 /d Blinatumomab steady state serum concentration at 15 µg/m 2 /d:.62 ng/ml (range ng/ml) In vitro EC 9 data predicted the human exposure required for anti-tumor efficacy 11

12 Ex vivo: Elimination of Lymphoma Cells by Blinatumomab Analogue Sample derived from B-CLL Patient with very low E:T cell ratio (1:48) Löffler et al. Leukemia 23 17:

13 Example: Ex vivo Elimination of AML Blasts by AMG 33 Kupka, C. et al. Blood 214 Jan 16;123(3):

14 Example: Ex Vivo Depletion of CD33 + Cells from Cyno Bone Marrow by AMG 33 CD33 + [% of viable cells] with AMG 33 w/o AMG 33 T-cells [% of all cells] with AMG 33 w/o AMG Time [h] Time [h] Friedrich, M. et al., Mol Cancer Ther. 214 Jun;13(6): (6) 14

15 In Vitro Pharmacology Package for FiH Study Continued For dose- and target cell-dependent T cell activation induction of surface markers for early and late T cell activation increase in T cell size and induction of proliferation specificity of T cell activation expression of granzyme and perforin in BiTE antibody-activated CD8 + and CD4 + T cells time- and dose-dependency of cytokine release by T cells 15

16 Example: In Vitro Induction of T Cell Proliferation by BLINCYTO TM CD19-depleted PBMC PBMC 1 Proliferation [%] MT12 PHA/ IL2 Blinatumomab MT13 Concentration [ng/ml] Amgen Report No. 13-PCD-65 16

17 Maximum Ensuing T Cell Expansion in Patients p = T Cells [1 3 / l] T Cells [1 3 / l] Baseline Maximum.. Baseline Klinger et al. Blood. 212;119:

18 Example: In Vitro Target Cell dependent Induction of Activation Markers on T Cells by BLINCYTO TM CD69- or CD25-positive T Cells [%] No Target Cells CHO (1%) CHO-CD19 (1%) CHO-CD19 (5%) CHO-CD19 (1%) CD CD CD CD Brischwein, K. et al. J Immunother. 27;3:

19 T Cell Activation Marker CD69 on CD8 + and CD4 + T Cells is Transiently Upregulated The T-cell activation marker CD69 was investigated for 6 patients and is transiently upregulated on CD8 + and CD4 + T cells after infusion start in cycle 1. 19

20 BLINCYTO TM induced Cytokine Release In Vitro Cytotox Assay with 1 µg/ml MT13 Cytotox Assay w/o MT13 Cytokine [pg/ml] IFN- TNF IL-1 IL-6 IL-4 IL-2 Cytokines 2

21 Cytokine Release by BLINCYTO TM is Transient and Modest Mean Peak Cytokine Levels in Serum of all Assessable Patients B Cycle 2 Mean Cytokine Concentration [pg/ml] 9 IL Time Point [Days] IL-6 IL-1 IFN- TNF- 28 Klinger et al. Blood. 212;119:

22 Cytokine Profiles of Individual Patients after 6 µg/m 2 /day BLINCYTO TM given as First Dose or After Double Dose Steps N= 21 N= µg/m 2 /day 6 µg/m 2 /day Note: Data < LOD were set to 1 pg/ml ( ½ LOD). Cytokine elevation was reduced after a stepwise dosing approach to a target dose of 6 µg/m 2 /day compared to a starting dose of 6 µg/m 2 /day 22

23 In Vivo: Activity of BLINCYTO TM Efficacy in a panel of xenograft models Tumor Volume [mm 3 ] 2 Gr.1 Vehicle w/o PBMC Treatment (days 1-5) Gr.2 Vehicle Gr.3 AMG 13 (.5 mg/kg) Gr.4 AMG 13 (.5 mg/kg) Gr.5 AMG 13 (.5 mg/kg) Gr.6 AMG 13 (.5 mg/kg) Survival [%] Treatment q1dx26 1 Vehicle control w/o T cells 2 Vehicle control 3 AMG 13 (.267 mg/kg/day); ** 4 AMG 13 (.27 mg/kg/day); ** 5 AMG 13 (.3 mg/kg/day); *** 6 AMG 13 (.133 mg/kg/day); *** Days after Tumor Cell Injection Day of study Raji Burkitt s Lymphoma Xenograft Granta-519 Mantle Cell Lymphoma Xenograft Amgen Report No. 13-PCD-98 and R

24 Peripheral B Cells are Depleted and T Cells Redistribute During BLINCYTO TM Treatment B- and T-cell counts varied widely between patients before infusion start. For all but one patient (nonresponder), peripheral B cells were depleted rapidly (< 1 week for most patients) and remained undetectable throughout treatment. There was a swift redistribution of T cells after treatment start (day 1) and dose step (day 8); T-cell counts recovered to baseline within several days, where they remained stable in most 24 patients.

25 Phase 2 Study with Blinatumomab in Patients with Residual B ALL in Bone Marrow Study Population B-precursor ALL patients in complete hematological remission with molecular failure or molecular relapse at any time after chemotherapy Leukemia Chemotherapy GMALL (Elderly) Protocol Blinatumomab 1-1 MRD pos MRD neg Induction Treatment 4 week continuous infusion Dose: 15 g/m²/d Cons. 1 Cons. 2 Cons. 3 Cons. 4 Patient #

26 Nonclinical Safety Evaluation

27 Toxicology Package for FiH Study Study of target-related risk TCR studies and RNA expression profiles Comparative target distribution between human and cyno tissue panels Comparative in vitro pharmacology in human and cyno co-culture systems incl. determination of MABEL Studies in non-human primates Exploratory dose range-finding in cyno monkeys Toxicity study in cyno monkeys including detailed hematology, toxicokinetics, immunogenicity and safety pharmacology assessments 27

28 Example : Determination of MABEL MABEL is the minimum anticipated biological effect level Effector cells: human PBMC (n=2) Selection of the Most Sensitive Test System Cell line with highest expression of TAA surface molecules Detemination of the most sensitive endpoint: redirected lysis T cell activation cytokine release Specific Lysis [%] MT112/BAY21112 [ng/ml] BiTE [ng/ml] Amgen Report No. 112-NCD-7 28

29 Common Effects Observed in Cynos within the First 48 hrs of Treatment Transient elevation of body temperature Transient cytokine release Hematologic changes transient reduction in LYMPH and MONO (recovered by Day 8) increased CD4 and CD8 cell activation (CD69) transient increase in NEUT Acute phase response elevated CRP, reduced albumin, increased globulin and fibrinogen 29

30 Example: PD Endpoints in Cyno Studies Blood FACS Day 7 Control BiTE -treated Group Prestudy Day 7 Bone Marrow FACS Bone marrow (sternum) 3

31 Cyno Model Relevant and Translates to Clinic Cyno is a pharmacologically-relevant model PD effects present indicators of BiTE activity observed signs of toxicity dependent on tumor-target expression on normal tissues BiTE effects in cyno mimic those observed clinically body temperature changes = fever/chills cytokine release hematological changes acute inflammatory response 31

32 Acknowledgements Amgen Matthias Friedrich Oliver Thomas Petra Deegen Anja Henn Alexander Sternjak Grit Lorenczewski Patrick A. Bäuerle Andreas Wolf and the ARM technical staff 32

33 AMGEN Research (Munich) GmbH 33