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Antibody-targeted chemotherapy of B-cell lymphoma using calicheamicin conjugated to murine or humanized antibody against CD22

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Abstract

Antibody-targeted chemotherapy with immunoconjugates of calicheamicin is a clinically validated strategy in cancer therapy. This study describes the selection of a murine anti-CD22 mAb, m5/44, as a targeting agent, its conjugation to a derivative of calicheamicin (CalichDM) via either acid-labile or acid-stable linkers, the antitumor activity of CalichDM conjugated to m5/44, and its subsequent humanization by CDR grafting. Murine IgG1 mAb m5/44 was selected based on its subnanomolar affinity for CD22 and ability to be internalized into B cells. CalichDM conjugated to m5/44 caused potent growth inhibition of CD22+ human B-cell lymphomas (BCLs) in vitro. The conjugate of m5/44 with an acid-labile linker was more potent than an acid-stable conjugate, a nonbinding conjugate with a similar acid-labile linker, or unconjugated CalichDMH in inhibiting BCL growth. CalichDM conjugated to m5/44 caused regression of established BCL xenografts in nude mice. In contrast, both unconjugated m5/44 and a nonbinding conjugate were ineffective against these xenografts. Based on the potent antitumor activity of m5/44-CalichDM conjugates, m5/44 was humanized by CDR grafting to create g5/44, an IgG4 anti-CD22 antibody. Both m5/44 and g5/44 bound CD22 with subnanomolar affinity. Competitive blocking with previously characterized murine anti-CD22 mAbs suggested that g5/44 recognizes epitope A located within the first N-terminal Ig-like domain of human CD22. Antitumor efficacy of CalichDM conjugated to g5/44 against BCL xenografts was more potent than its murine counterpart. Based on these results, a calicheamicin conjugate of g5/44, CMC-544, was selected for further development as a targeted chemotherapeutic agent for the treatment of B-cell malignancies.

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Abbreviations

AcBut:

4-(4′-Acetylphenoxy) butanoic acid

AcPAc:

(3-Acetylphenyl) acetic acid

ATC:

Antibody-targeted chemotherapy

BCL:

B-cell lymphoma

CalichDM:

N-Acetyl-γ-calicheamicin dimethyl disulfide derivative(s)

CalichDMA:

CalichDM acid

CalichDMH:

CalichDM hydrazide

CDR:

Complementarity determining region

NHL:

Non-Hodgkin’s lymphoma

PBMC:

Peripheral blood mononuclear cell

TAA:

Tumor-associated antigen

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Acknowledgements

We would like to thank Dr John Crocker of Birmingham Heartlands Hospital, Birmingham, UK, for the evaluation of m5/44 binding to the NHL biopsies; Dr Lyka Kalyandrug for studies with effector functions of antibodies; Fred Immerman for statistical evaluation of results; and Maureen Dougher and Latha Sridharan for assistance with various studies.

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Authors and Affiliations

  1. Oncology Discovery, Wyeth Research, 200/4604, 401 North Middletown Road, Pearl River, NY, 10965, USA

    John F. DiJoseph, Kiran Khandke, Douglas C. Armellino, Erwin R. Boghaert & Nitin K. Damle

  2. Chemical Sciences, Wyeth Research, Pearl River, New York, USA

    Arthur Kunz & Philip R. Hamann

  3. Celltech R&D, Slough, UK

    Andrew Popplewell, Simon Tickle, Heather Ladyman, Alastair Lawson, Karen Zinkewich-Peotti, Sue Stephens & Neil Weir

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  1. John F. DiJoseph
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  2. Andrew Popplewell
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Correspondence to Nitin K. Damle.

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DiJoseph, J.F., Popplewell, A., Tickle, S. et al. Antibody-targeted chemotherapy of B-cell lymphoma using calicheamicin conjugated to murine or humanized antibody against CD22. Cancer Immunol Immunother 54, 11–24 (2005). https://doi.org/10.1007/s00262-004-0572-2

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  • DOI: https://doi.org/10.1007/s00262-004-0572-2

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