1. Jakub Golab Department of Immunology Medical University of Warsaw
Therapeutic applications of anti-CD20 monoclonal antibodies - tumors and beyond
CHMP/PDCO/CMDh Presidency meeting
Jakub Golab
Department of Immunology
Medical University of Warsaw

2. Nobel prize in 1901 Emil Adolf von Behring Marburg University
"for his work on
serum therapy…”

4. Antigen recognition
antigen
antibody

5. Nobel prize in 1984 "for the discovery of the principle for production
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"

6. Types of monoclonal antibodies
murine Fab
murine FV Fc
Fab
murine antibody
chimeric antibody
(66% of human sequence)
humanised antibody
(90% of human sequence)
human antibody
(100% of human sequence)

8. Monoclonal antibodies approved for clinical use in oncology
mAb name
Target antigen
Indication
Clinical approval
Rituximab
CD20
Non-Hodgkin lymphoma
1997
Ibritumomab tiuxetan
2002
Tositumomab
2003
Ofatumumab
Chronic lymphocytic leukemia
2009
Alemtuzumab
CD52
2001
Gemtuzumab ozogamicin
CD33
Acute myelogenous leukemia
2000
Bevacizumab
VEGF
Colorectal cancer
Non-small cell lung cancer
Breast cancer
Glioblastoma
Kidney cancer
2004
2006
2008
Panitumumab
EGFR
Cetuximab
Head & neck cancers
Nimotuzumab
2005
Trastuzumab
HER2
1998
Ipilimumab
CTLA-4
Malignant melanoma
2011
Brentuximab vedotin
CD30
Hodgkin lymphoma
Edrecolomab
EpCAM
1995

9. Mechanisms of mAbs action (1):
Winiarska et al. Frontiers Biosci 2011;16:277

10. Mechanisms of mAbs action (2):
Winiarska et al. Frontiers Biosci 2011;16:277

11. Structure of CD20 antigen
Magdalena Winiarska

12. Anti-CD20 monoclonal antibodies
Antibody
Clinical status
Characteristics
Origin
Isotype
Rituximab
approved
chimeric
IgG1
Tositumomab*
murine
IgG2a
Ofatumumab
human
Ibritumomab**
Ocrelizumab
phase 3
humanized
Veltuzumab
phase 2
Obinutuzumab
PRO131921
AME-133
LFB-R603/EMAB-6
phase 1
* radioimmunoconjugate bound to 131iodine
** radioimmunoconjugate bound to 90ytrium

13. Development of novel anti-CD20 monoclonal antibodies:

14. Rituximab-mediated complement dependent cytotoxicity against control and statin-treated lymphoma cells
Winiarska et al. PLoS Med. 2008;5(3):e64.

15. and why this finding might be important … ?
Because 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

16. Number of cells
Fluorescence intensity
Winiarska et al. PLoS Med. 2008;5(3):e64.

17. PE – intracellular tail of CD20 FITC – extracellular loop of CD20
Winiarska et al. PLoS Med. 2008;5(3):e64.

18. What is the mechanism?

19. Farnesyltransferaze inhibitor (L-744,832) increases CD20 expression
CD20 promoter activity
24 h
48 h
Western blotting
Flow cytometry
CHIP assay
Surface CD20
Magda Winiarska

20. L-744,832 [μM] 1 10 100 rituximab [µg/ml] 40 80
Farnesyltransferaze inhibitor (L-744,832) potentiates rituximab-mediated CDC 1 10
100
rituximab [µg/ml] 40 80
L-744,832 [μM]
Survival [% of controls]
Magda Winiarska

21. Cancer patients are unlikely to give up these treatments!!!
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)
Cancer patients are unlikely to
give up these treatments!!!
Elderly people usually take multiple different drugs:
antidiabetics
statins
diuretics
β-blockers
antidepressants
anti-inflammatory drugs

22. Baker, Nature Biotech 23, 1065 (2005)

23. 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.

24. 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. Mackus
Patrick J. Engelberts
Paul 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
Anna Dąbrowska-Iwanicka
Maria Sklodowska-Curie Memorial Cancer Center
Krzysztof Warzocha
Institute of Hematology, Warsaw
Kazimierz Sułek
Department of Clinical Hematology, Military Medical Institute
Some figures were prepared using Servier Medical Art (

25. T E A M 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