therapeutic applications of anti- cd20 monoclonal antibodies - tumors and beyond jakub golab...
TRANSCRIPT
Therapeutic applications of anti-CD20 monoclonal antibodies - tumors and beyond
Jakub GolabDepartment of Immunology
Medical University of Warsaw
CHMP/PDCO/CMDh Presidency meeting
Emil Adolf von BehringMarburg University
Nobel prize in 1901
"for his work onserum therapy…”
antigen antibody
Antigen recognition
Georges KöhlerBasel Institute for
Immunology
César MilsteinMRC Laboratory of Molecular Biology
Cambridge
Nobel prize in 1984
"for the discovery of the principle for production of monoclonal antibodies"
Types of monoclonal antibodies
murine antibody chimeric antibody(66% of human
sequence)
humanised antibody
(90% of human sequence)
human antibody(100% of human
sequence)
murine Fab
murine FV
Fab Fc
mAb name Target antigen Indication Clinical approval
Rituximab CD20 Non-Hodgkin lymphoma 1997Ibritumomab tiuxetan CD20 Non-Hodgkin lymphoma 2002
Tositumomab CD20 Non-Hodgkin lymphoma 2003Ofatumumab CD20 Chronic lymphocytic leukemia 2009Alemtuzumab CD52 Chronic lymphocytic leukemia 2001Gemtuzumab ozogamicin CD33 Acute myelogenous leukemia 2000
Bevacizumab VEGF Colorectal cancerNon-small cell lung cancerBreast cancerGlioblastomaKidney cancer
20042006200820092009
Panitumumab EGFR Colorectal cancer 2006Cetuximab EGFR Colorectal cancer
Head & neck cancers20042006
Nimotuzumab EGFR Head & neck cancers 2005Trastuzumab HER2 Breast cancer 1998Ipilimumab CTLA-4 Malignant melanoma 2011Brentuximab vedotin CD30 Hodgkin lymphoma 2011Edrecolomab EpCAM Colorectal cancer 1995
Monoclonal antibodies approved for clinical use in oncology
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
Antibody Clinical status
Characteristics
Origin Isotype
Rituximab approved chimeric IgG1Tositumomab* approved murine IgG2aOfatumumab approved human IgG1Ibritumomab** approved murine IgG1Ocrelizumab phase 3 humanized IgG1Veltuzumab phase 2 humanized IgG1Obinutuzumab phase 2 humanized IgG1PRO131921 phase 2 humanized IgG1AME-133 phase 2 humanized IgG1LFB-R603/EMAB-6 phase 1 chimeric IgG1
* radioimmunoconjugate bound to 131iodine ** radioimmunoconjugate bound to 90ytrium
Anti-CD20 monoclonal antibodies
Development of novel anti-CD20 monoclonal antibodies:
Su
rviv
al [
% o
f co
ntr
ols
]
80
100
60
40
20
0
lov
a 1
µM
lov
a 5
µM
lov
a 1
0 µM
*
*lo
va
0 µ
M
+ R itux+ AB serum
sim
va
me
va
flu
va
ato
rva
**
**
n o s ta tinstatin
lov
a
*
ce
riv
a
*
pra
va
*
Rituximab-mediated complement dependent cytotoxicity against control and
statin-treated lymphoma cells
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 long-term statin users
• The estimated number of people that should be
using statins at the time being is 200 million
Co
ntr
ols
12
h
Co
ntr
ols
48
h
Lo
va 1
h
Lo
va 1
2 h
Lo
va 2
4 h
Lo
va 4
8 h
CD20Actin
R T-PC R
dH
O2
CD20Tubulin
Co
ntr
ols
Lo
va
5 µ
M
Lo
va
10 µ
M
Lo
va
20 µ
M
Lo
va
30 µ
M
W B
Num
ber o
f cel
ls
Fluorescence intensity
0
32
0.1 1000S
urv
iva
l [%
of
con
tro
ls]
80
60
40
20
0
ctrl lova lova+
chol
0 0 .5 1 .0 5 .0 10 .0
M ßC D [m g/m l]
Winiarska et al. PLoS Med. 2008;5(3):e64.
PE – intracellular tail of CD20FITC – extracellular loop of CD20
Controls Lova 10
Winiarska et al. PLoS Med. 2008;5(3):e64.
What is the mechanism?
1 10
L concentration [µM ]
50
1000
800
600
400
200
0
CD
20 p
rom
ote
r a
ctiv
ity
[% o
f c
on
tro
ls]
Raji - pCD20luc-2-w t
0 L5 L10 L200
100
200
300
CD20
Tubulin
Co
ntr
ols
L 5
µM
L 1
0 µ
M
L 2
0 µ
M
Su
rfac
e C
D20
24 h48 h
Magda Winiarska
Farnesyltransferaze inhibitor (L-744,832) increases CD20 expression
Western blotting Flow cytometry
CD20 promoter activity
CHIP assay
0
1 0
2 0
L [µM]
Su
rviv
al [
% o
f c
on
tro
ls]
1 10 100rituximab [µg/ml]
0
40
80
L-744,832 [μM]
Magda Winiarska
Farnesyltransferaze inhibitor (L-744,832) potentiates rituximab-mediated CDC
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)
• Increased number of comorbid diseases (cardiovascular diseases, diabetes, rheumatic diseases,
neuropsychiatric disorders)
Elderly people usually take multiple different drugs:
• antidiabetics• statins• diuretics• β-blockers
• antidepressants• anti-inflammatory drugs
Cancer patients are unlikely to give up these treatments!!!
Baker, Nature Biotech 23, 1065 (2005)
H S CCD 34 +
p ro -B
C D 2 0CD 19
IgM
+
-
-
p re -BCD 19
IgM
+
-C D 2 0 +
im m a tu re B c e llCD 19
IgM
+
+C D 2 0 +
n a v e B c ellϊCD 19
CD 38
+
+/-
C D 2 0 +
IgM +g e rm in a l c e n ter
B c e llCD 19
CD 38
+
++
C D 2 0 +
IgM +
m e m o ry B c e llCD 19 +
C D 2 0 +
IgM /IgG /IgA /IgE +
p la s m a c e llCD 19
CD 38
+/-
+++
C D 2 0 -
IgM
CD 138
-
+
B O N E M A R R O W LY M P H N O D E
C IR C U L AT IO N
Potential future uses of anti-CD20 monoclonal antibodies
rheumatoid arthritismultiple sclerosis
systemic lupus erythematosusautoimmune anemia
autoimmune hemolytic anemiapure red cell aplasia
idiopathic thrombocytopenic purpura (ITP)Wegener's granulomatosis
bullous skin disorders (pemphigus, pemphigoid)type 1 diabetes mellitus
Sjogren's syndromethyroid-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.
Some figures were prepared using Servier Medical Art (www.servier.com)
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
Department of Immunology
Medical University of Warsaw
Cezary Wójcik Department of Anatomy and Cell Biology, IUSM, USA
Grzegorz M . Wilczyński Nencki Institute of Experimental Biology
Wendy J.M. MackusPatrick J. EngelbertsPaul W.H.I. Parren
Genmab, Utrecht, The Netherlands
Łukasz Bojarski International Institute of Molecular and Cell Biology, Warsaw
Maciej Siński, Ewa Wilczek, Grzegorz Basak, Zbigniew Gaciong, Elżbieta Górska, Maria Wąsik
Medical University of Warsaw
Anna Dąbrowska-Iwanicka Maria Sklodowska-Curie Memorial Cancer Center
Krzysztof Warzocha Institute of Hematology, Warsaw
Kazimierz Sułek Department of Clinical Hematology, Military Medical Institute
T E A MImprovement 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 FirczukDr. Magdalena WiniarskaMSc. Małgorzata JodłowskaMSc. Angelika SzokalskaBSc. Justyna ChlebowskaBSc. Paweł SalwaBSc. Kamil BojarczukBSc. Magdalena Gabrysiak
TEAM coordinator:
prof. Jakub Golab
TEAM consultant:
Dr. Dominika Nowis