Transcript Anticancer monoclonal antibodies

Therapeutic applications of antiCD20 monoclonal antibodies tumors and beyond
CHMP/PDCO/CMDh Presidency meeting
Jakub Golab
Department of Immunology
Medical University of Warsaw
Nobel prize in 1901
Emil Adolf von Behring
Marburg University
"for his work on
serum therapy…”
Antigen recognition
antigen
antibody
Nobel prize in 1984
Georges Köhler
Basel Institute for
Immunology
César Milstein
MRC Laboratory of
Molecular Biology
Cambridge
"for the discovery of
the principle for production
of monoclonal antibodies"
Types of monoclonal antibodies
murine Fab
Fc
murine antibody
murine FV
Fab
chimeric
antibody
(66% of human
sequence)
humanised antibody
(90% of human
sequence)
human
antibody
(100% of
human
sequence)
Monoclonal antibodies approved for clinical use in oncology
mAb name
Target antigen
Indication
Clinical
approval
1997
2002
Rituximab
Ibritumomab tiuxetan
CD20
CD20
Non-Hodgkin lymphoma
Non-Hodgkin lymphoma
Tositumomab
Ofatumumab
Alemtuzumab
Gemtuzumab ozogamicin
CD20
CD20
CD52
CD33
Non-Hodgkin lymphoma
Chronic lymphocytic leukemia
Chronic lymphocytic leukemia
Acute myelogenous leukemia
2003
2009
2001
2000
Bevacizumab
VEGF
Panitumumab
Cetuximab
EGFR
EGFR
Colorectal cancer
Non-small cell lung cancer
Breast cancer
Glioblastoma
Kidney cancer
Colorectal cancer
Colorectal cancer
Head & neck cancers
Head & neck cancers
Breast cancer
Malignant melanoma
Hodgkin lymphoma
Colorectal cancer
2004
2006
2008
2009
2009
2006
2004
2006
2005
1998
2011
2011
1995
Nimotuzumab
Trastuzumab
Ipilimumab
Brentuximab vedotin
Edrecolomab
EGFR
HER2
CTLA-4
CD30
EpCAM
Mechanisms of mAbs action (1):
Winiarska et al. Frontiers Biosci 2011;16:277
Mechanisms of mAbs action (2):
Winiarska et al. Frontiers Biosci 2011;16:277
Structure of CD20 antigen
Magdalena Winiarska
Anti-CD20 monoclonal antibodies
Characteristics
Antibody
Clinical
status
Origin
Isotype
Rituximab
Tositumomab*
Ofatumumab
Ibritumomab**
Ocrelizumab
Veltuzumab
Obinutuzumab
PRO131921
AME-133
LFB-R603/EMAB-6
approved
approved
approved
approved
phase 3
phase 2
phase 2
phase 2
phase 2
phase 1
chimeric
murine
human
murine
humanized
humanized
humanized
humanized
humanized
chimeric
IgG1
IgG2a
IgG1
IgG1
IgG1
IgG1
IgG1
IgG1
IgG1
IgG1
* radioimmunoconjugate bound to 131iodine
** radioimmunoconjugate bound to 90ytrium
Development of novel anti-CD20 monoclonal
antibodies:
100
*
*
80
*
*
60
*
*
*
*
*
40
+ Ritux
+ AB serum
prava
ceriva
lova
atorva
fluva
meva
lova 10 µM
lova 5 µM
lova 1 µM
0
simva
20
lova 0 µM
Survival [% of controls]
Rituximab-mediated complement dependent
cytotoxicity against control and statin-treated
lymphoma cells
no statin
statin
Winiarska et al. PLoS Med. 2008;5(3):e64.
and why this finding might be
important … ?
• Because 30-40 million people
all over the world are longterm statin users
• The estimated number of
people that should be using
statins at the time being is
200 million
Number of cells
Lova 30 µM
Lova 20 µM
Lova 5 µM
Controls
dH2O
Lova 48 h
Lova 24 h
Lova 12 h
Lova 1h
Controls 48 h
Controls 12h
CD20
Actin
Lova 10 µM
WB
RT-PCR
CD20
Tubulin
32
0
0.1
1000
Fluorescence intensity
Survival [% of controls]
80
60
40
20
0
ctrl
lova
lova
+
chol
0
0.5
1.0
5.0
10.0
MßCD [mg/ml]
Winiarska et al. PLoS Med. 2008;5(3):e64.
PE – intracellular tail of CD20
FITC – extracellular loop of CD20
Controls
Lova 10
Winiarska et al. PLoS Med. 2008;5(3):e64.
What is the mechanism?
Farnesyltransferaze inhibitor (L-744,832) increases
CD20 expression
Raji
- pCD20luc-2-wt
CD20
promoter
activity
CD20 promoter activity
[% of controls]
1000
800
24 h
48 h
600
400
200
0
0
1
5
10
L concentration [µM]
Flow cytometry
Tubulin
L 20 µM
L 10 µM
Surface CD20
CD20
CHIP assay
300
L 5 µM
Controls
Western blotting
200
100
0
0
L5
L10
L20
Magda Winiarska
Survival [% of controls]
Farnesyltransferaze inhibitor (L-744,832) potentiates
rituximab-mediated CDC
80
L [µM] [μM]
L-744,832
0
10
20
40
0
1
10
100
rituximab [µg/ml]
Magda Winiarska
Cancer is a disease of elderly people
Increasing age is associated with:
• complex changes in physiology (alterations in renal and
hepatic functions, decreased bone marrow reserve)
Cancer patients are unlikely to
• Increased
number
of comorbid
diseases (cardiovascular
give
up these
treatments!!!
diseases, diabetes, rheumatic diseases,
neuropsychiatric disorders)
Elderly people usually take multiple different drugs:
• antidiabetics
• statins
• diuretics
• β-blockers
• antidepressants
• anti-inflammatory drugs
BONE MARROW
LYMPH NODE
pre-B
+
CD19
+
CD20
IgM
immature B cell
+
CD19
+
CD20
+
IgM
pro-B
CD19+
CD20IgM
HSC
+
CD34
plasma cell
CD19+/CD20IgM
+++
CD38
+
CD138
naϊ ve B cell
+
CD19
+
CD20
+
IgM
+/CD38
germinal center
B cell
CD19+
CD20+
+
IgM
++
CD38
memory B cell
CD19+
CD20+
+
IgM/IgG/IgA/IgE
CIRCULATION
Baker, Nature
Biotech 23, 1065 (2005)
Potential future uses of anti-CD20
monoclonal antibodies
rheumatoid arthritis
multiple sclerosis
systemic lupus erythematosus
autoimmune anemia
autoimmune hemolytic anemia
pure red cell aplasia
idiopathic thrombocytopenic purpura (ITP)
Wegener's granulomatosis
bullous skin disorders (pemphigus, pemphigoid)
type 1 diabetes mellitus
Sjogren's syndrome
thyroid-associated ophthalmopathy
A new study from Norway suggests that rituximab (together with
methotrexate) might help patients with chronic fatigue syndrome. A
clinical trial is ongoing.
Acknowledgements
Magdalena Winiarska, Jacek Bil, Dominika Nowis,
Małgorzata Wańczyk, Marcin Makowski, Tomasz Świtaj,
Kamil Bojarczuk, Eliza Głodkowska, Piotr Mrówka,
Paweł Salwa, Tadeusz Issat, Zuzanna Kurzaj, Witold
Lasek, Tomasz Stokłosa, Marek Jakóbisiak
Cezary Wójcik
Grzegorz M . Wilczyński
Wendy J.M. Mackus
Patrick J. Engelberts
Paul W.H.I. Parren
Łukasz Bojarski
Maciej Siński, Ewa Wilczek, Grzegorz Basak, Zbigniew
Gaciong, Elżbieta Górska, Maria Wąsik
Anna Dąbrowska-Iwanicka
Krzysztof Warzocha
Kazimierz Sułek
Department of Immunology
Medical University of Warsaw
Department of Anatomy and Cell
Biology, IUSM, USA
Nencki Institute of Experimental Biology
Genmab, Utrecht, The Netherlands
International Institute of Molecular and
Cell Biology, Warsaw
Medical University of Warsaw
Maria Sklodowska-Curie Memorial
Cancer Center
Institute of Hematology, Warsaw
Department of Clinical Hematology,
Military Medical Institute
Some figures were prepared using Servier Medical Art (www.servier.com)
TEAM
Improvement of antitumor effectiveness of photodynamic therapy
In coooperation with:
• Patrizia Agostinis (Catholic University of Leuven, Belgium)
• Michael Hamblin (Wellmans Center of Photomedicine, Boston, USA)
• Jacek Capala (National Institutes of Health, Bethesda, USA)
TEAM members:
Dr. Małgorzata Firczuk
Dr. Magdalena Winiarska
MSc. Małgorzata Jodłowska
MSc. Angelika Szokalska
BSc. Justyna Chlebowska
BSc. Paweł Salwa
BSc. Kamil Bojarczuk
BSc. Magdalena Gabrysiak
TEAM coordinator:
prof. Jakub Golab
TEAM consultant:
Dr. Dominika Nowis