GAZYVA®/ GAZYVARO™ The Success Story of a Glycoengineered Antibody CMC Strategy Forum Japan 2014 December 8-9, 2014 Dr. Elisabeth Kirchisner Technical Regulatory Affairs Roche Diagnostics GmbH, Germany
The Molecule
Obinutuzumab Tradename
GAZYVA® (US, other countries) GAZYVARO™ (EU, Switzerland)
INN, other names
Obinutuzumab, GA101
Molecule Type
Recombinant, humanized, monoclonal Type II glycoengineered anti-CD20 antibody (IgG1)
Indications
• chronic lymphocytic leukemia (CLL) • indolent non-Hodgkin’s lymphoma (iNHL) • diffuse large B-cell lymphoma (DLBCL)
Start of clinical trials
Sept 2007
First market approvals (CLL)
01 Nov 2013 (US) Approved in EU, Switzerland, Canada, Australia, South Korea, others 2
Obinutuzumab Molecule Obinutuzumab is a Glycoengineered Type II Antibody with a Unique Mode of Action
Type II anti-CD20 antibody1
Glycoengineered Fc region2
Enhanced direct cell death
Increased antibodydependent cellular cytotoxicity (ADCC)
1
Niederfellner G, et al. Blood 2011; 118:358–367. 2. Alduaij W, et al. Blood 2011; 117:4519–4529.
2
Mössner E, et al. Blood 2010; 115:4393‒4402. 4. Herter S, et al. Blood 2010; 116:Abstract 3925.
3
Glycoengineering (GlycoMAb™ Technology)
• Co-transfection of genes encoding for the antibody with genes encoding for glycosylation-modifying enzymes • Glycosylation-modifying enzymes:
– GnTIII (N-acetylglucosaminyltransferase III) – ManII (α-Mannosidase II) • Co-expression of the antibody with glycosylation-modifying enzymes during cell culture Modified glycosylation pattern with reduced levels of core-fucosylation Increase in antibody-dependent cell-mediated cytotoxicity (ADCC)
4
Biosynthesis of Glycosylation - Pathway
Glycostructures – “Normal antibody” G0
G0-Fuc G1-Fuc
G1*
Man5 G2
G0-GlcNac
Gal2-NANA
G1, α-1-3 linked antenna (* isobaric structures) G1, α-1-6 linked antenna
Glycostructures – “Glycoengineered Antibody” bG0-Fuc
bG0
bG1-Fuc
bG1
hyb_bMan5G0-Fuc
Isobaric structures: Hyb_bMan4G1-Fuc
G2-Fuc
Obinutuzumab Exhibits up to 100-fold Higher ADCC Potency than Rituximab and Ofatumumab GA101 Z138 (PBMC: V/V)
100
Antibody-dependent killing (%)
Rituximab
Ofatumumab
100
80
80
60
60
40
40
20
20
0
0
–20
0.064 0.32 1.6
8
40
200 1000
Antibody concentration (ng/ml)
Herter et al., MCT, 2013
–20
SU-DHL4 (NK92 LC3 E11)
0.0001 0.001 0.01 0.1
1
10
100
Antibody concentration (ng/ml)
Obinutuzumab Mediates Increased Direct Cell Death Induction on a Panel of NHL Cell Lines
% of cells (AnnexinV+ and Annexin V+/PI+)
80
Untreated 10 µg/ml GA101 10 µg/ml rituximab
100
70 80
60
50
60
40 30
40
20
20
10 0
0
Annexin/PI assay
Mössner et al., Blood , 2010; Herter et al. Mol Canc Ther, 2013
Direct cell death
Untreated CPT GA101 Rituximab Ofatumumab
Herter, et al. Mol Canc Ther, 2013 Z138 cells, AxV: green, PI: red, 0-6 h
Obinutuzumab Versus Rituximab in CLL: Progression-free Survival
Progression-free survival
0.9 0.8 0.7 0.6
Patients with a response (%)
G-CIb R-CIb Stratified HR: 0.39 95% CI: 0.31–0.49 p
