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Development of CDX-1140, an agonist CD40 antibody for cancer immunotherapy

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Abstract

Limitations of immunotherapy include poorly functioning events early in the immune response cycle, such as efficient antigen presentation and T cell priming. CD40 signaling in dendritic cells leads to upregulation of cell surface costimulatory and MHC molecules and the generation of cytokines, which promotes effective priming of CD8+ effector T cells while minimizing T cell anergy and the generation of regulatory T cells. This naturally occurs through interaction with CD40 ligand (CD40L) expressed on CD4+ T-helper cells. CD40 signaling can also be achieved using specific antibodies, leading to several agonist CD40 antibodies entering clinical development. Our approach to select a CD40 agonist antibody was to define a balanced profile between sufficiently strong immune stimulation and the untoward effects of systemic immune activation. CDX-1140 is a human IgG2 antibody that activates DCs and B cells and drives NFkB stimulation in a CD40-expressing reporter cell line. These activities are Fc-independent and are maintained using an F(ab′)2 fragment of the antibody. CDX-1140 binds outside of the CD40L binding site, and addition of recombinant CD40L greatly enhances DC and B activation by CDX-1140, suggesting that CDX-1140 may act synergistically with naturally expressed CD40L. CDX-1140 also has both direct and immune-mediated anti-tumor activity in xenograft models. CDX-1140 does not promote cytokine production in whole blood assays and has good pharmacodynamic and safety profiles in cynomolgus macaques. These data support the potential of CDX-1140 as part of a cancer therapy regimen, and a phase 1 trial has recently commenced.

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Abbreviations

AAALAC:

Association for Assessment and Accreditation of Laboratory Animal Care International

CD40L:

CD40 ligand also referred to as CD154

CHO:

Chinese hamster ovary

CRD1:

Cysteine-rich domain 1

CRD2:

Cysteine-rich domain 2

CRD3:

Cysteine-rich domain 3

CRD4:

Cysteine-rich domain 4

Cyno:

Cynomolgus macaque

ECD:

Extracellular domain

Hu:

Human

MSD:

Meso Scale Discovery

MTD:

Maximum tolerated dose

NOAEL:

No observable adverse effect level

rCD40L:

Recombinant CD40L

TMB:

3,3′,5,5′-tetramethylbenzidine

TRAF:

TNF receptor associated factors

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Acknowledgements

The authors would like to thank Mallary L. Rocheleau, Michelle E. Grealish, Catherine D. Pilsmaker, Elizabeth Q. Do, Kathleen M. Borrelli, James Testa, Laura Mills-Chen, Collen Patterson and Karla Keler for expert technical assistance.

Funding

This work was funded by Celldex Therapeutics, Inc.

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Authors and Affiliations

  1. Celldex Therapeutics, Inc, 53 Frontage Road, Suite 220, Hampton, NJ, 08827, USA

    Laura A. Vitale, Li-Zhen He, Thomas O’Neill, Jenifer Widger, Andrea Crocker, Karuna Sundarapandiyan, Jeffrey Weidlick, Crystal Sisson, Joel Goldstein & Tibor Keler

  2. Celldex Therapeutics, Inc, Needham, MA, USA

    Lawrence J. Thomas, James R. Storey, Eric M. Forsberg, April R. Baronas, Lauren E. Gergel, James M. Boyer & Henry C. Marsh Jr.

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  1. Laura A. Vitale
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Contributions

Laura A. Vitale: study design, methodology, data analysis, and manuscript preparation. Lawrence J. Thomas: study design, methodology, data analysis, and manuscript preparation. Li-Zhen He: study design, methodology, data analysis, and manuscript preparation. Thomas O’Neill: experimental design, methodology, and performance; data analysis. Jenifer Widger: recombinant DNA design, construction and characterization. Andrea Crocker: experimental design, methodology, and performance; data analysis. Karuna Sundarapandiyan: experimental design, methodology, and performance; data analysis. James R. Storey: recombinant DNA design, construction and characterization. Eric M. Forsberg: experimental design, methodology, and performance; data analysis. Jeffrey Weidlick: experimental design, methodology, and performance; data analysis. April R. Baronas: experimental design, methodology, and performance; data analysis. Lauren E. Gergel: experimental design, methodology, and performance; data analysis. James M. Boyer: experimental design, methodology, and performance; data analysis. Crystal Sisson: experimental design, methodology, and performance; data analysis. Joel Goldstein: study design, data analysis, and manuscript preparation. Henry C. Marsh, Jr.: study design, data analysis, and manuscript preparation. Tibor Keler: study design, data analysis, and manuscript preparation.

Corresponding author

Correspondence to Tibor Keler.

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Conflict of interest

All authors are employees of, and own stock or stock option in Celldex Therapeutics, Inc.

Ethical approval

and ethical standards and animal sources.

Animals were sourced from IACUC-approved commercial sources. Murine xenograft studies (animal source: Taconic Biosciences) were approved by the Celldex IACUC of Hampton, NJ (AUP CDX-002) or the Celldex IACUC of Needham, MA (AUP 08-2017). Animal care followed the Guide for the Care and Use of Laboratory Animals: Eighth Edition (National Research Council. 2011. Washington, DC: The National Academies Press). The pilot primate study (animal source: Charles River Laboratories) was approved by the Charles River Laboratories IACUC of Shrewsbury, MA (AUP 20097548). Those animals were handled according to Guide for the Care and Use of Laboratory Animals: Eighth Edition and AAALAC rules (Association for Assessment and Accreditation of Laboratory Animal Care International). The primate toxicology study (animal source: Kunming Biomed International LTD) was approved by the Citoxlab IACUC of Montreal, QC (AUP 1016–3273). Those animals were handled according to The Canadian Council on Animal Care and AAALAC rules.

Cell line authentication.

Cell lines were sourced directly from vendors that provide authentication. CHO and EJ138 cells were purchased from Millipore-Sigma, HEK-293 cells were purchased from InvivoGen, NFkB luciferase reporter HEK293 stable cell line was purchased from Signosis, and the Ramos and Raji cell lines were purchased from ATCC.

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Vitale, L.A., Thomas, L.J., He, LZ. et al. Development of CDX-1140, an agonist CD40 antibody for cancer immunotherapy. Cancer Immunol Immunother 68, 233–245 (2019). https://doi.org/10.1007/s00262-018-2267-0

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