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:

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n o s ta tinstatin

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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

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h

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CD20Actin

R T-PC R

dH

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CD20Tubulin

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of

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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?

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L concentration [µM ]

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20 p

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[% o

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0 L5 L10 L200

100

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Tubulin

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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]

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1 10 100rituximab [µg/ml]

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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