Compositions and methods for treating cancer with anti-CD33 immunotherapy
Inventors
ORENTAS, Rimas • Schneider, Dina • Dropulic , Boro • Dimitrov, Dimiter S. • Zhu, Zhongyu
Assignees
Lentigen Technology Inc • US Department of Health and Human Services
Publication Number
US-10426797-B2
Publication Date
2019-10-01
Expiration Date
2038-03-23
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Abstract
Chimeric antigen receptors containing CD33 antigen binding domains are disclosed. Nucleic acids, recombinant expression vectors, host cells, antigen binding fragments, and pharmaceutical compositions, relating to the chimeric antigen receptors are also disclosed. Methods of treating or preventing cancer in a subject, and methods of making chimeric antigen receptor T cells are also disclosed.
Core Innovation
The invention discloses chimeric antigen receptors (CARs) containing CD33 antigen binding domains, as well as related nucleic acids, recombinant expression vectors, host cells (including T cells) expressing the receptors, antigen binding fragments, and pharmaceutical compositions. The CARs exhibit high surface expression on transduced T cells, demonstrate a high degree of cytolysis, and show transduced T cell in vivo expansion and persistence. The invention also includes methods of making CAR T cells and methods of treating or preventing cancer in a subject using these CARs.
The background describes the critical need for improved treatments for acute myeloid leukemia (AML) and other cancers expressing CD33, due to limited survival benefits with current treatments and toxicity associated with existing CD33-targeting therapies. CD33 is a myeloid lineage differentiation antigen highly expressed on AML cells but less so on normal differentiated myeloid cells. Although several anti-CD33 antibody-based therapies and CAR T cells are under development, a number of these approaches face clinical toxicity and suboptimal efficacy. The invention addresses these challenges by providing CAR compositions with fully human anti-CD33 antigen binding domains, which may reduce immunogenicity and improve CAR T cell persistence and function.
The summary highlights that the present invention provides novel anti-CD33 antibodies and antigen binding fragments, CARs containing these domains, and host cells expressing them. These CARs induce high surface expression and cytolytic activity on transduced T cells, alongside increased in vivo expansion and persistence. The disclosure includes nucleic acid sequences encoding these CARs and methods of using them for cancer treatment, particularly targeting diseases with dysregulated CD33 expression.
Claims Coverage
The patent includes multiple independent claims focusing on isolated nucleic acid molecules encoding CARs, vectors comprising these nucleic acids, cells transformed with these vectors, pharmaceutical compositions containing CAR T cells expressing the CARs, and methods of treating cancers expressing CD33 using these CAR T cells.
Chimeric antigen receptor with specific structural domains
An isolated nucleic acid molecule encoding a CAR comprising, from N-terminus to C-terminus: (i) at least one extracellular antigen binding domain comprising a CD33 antigen binding domain encoded by specific nucleotide sequences (SEQ ID NOs: 1, 3, 5, 7, 9, or 11); (ii) a transmembrane domain comprising a transmembrane domain from selected T-cell receptor or co-stimulatory proteins (e.g., TCR alpha, beta or zeta chain; CD28; CD3 epsilon; CD45; CD4; CD5; CD8; CD9; CD16; CD22; CD33; CD37; CD64; CD80; CD86; CD134; CD137; CD154; or combinations thereof); (iii) at least one costimulatory domain comprising functional signaling domains selected from OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1, ICOS, DAP10, DAP12, and 4-1BB, or combinations thereof; and (iv) an intracellular signaling domain comprising functional domains selected from 4-1BB, CD28, and CD3 zeta signaling domain, or combinations thereof.
Vectors incorporating the nucleic acid molecule encoding the CAR
Vectors comprising the isolated nucleic acid molecules encoding the CAR, which can be DNA, RNA, plasmid, cosmid, herpes virus, measles virus, lentivirus, adenoviral, retrovirus, or combinations thereof.
Cells expressing the CAR
Cells comprising the vector containing the nucleic acid molecule encoding the CAR, particularly human T cells transduced with the vector.
Pharmaceutical compositions containing CAR T cells
Pharmaceutical compositions comprising an anti-tumor effective amount of a population of human T cells transduced with the vector encoding the CAR, wherein the T cells are of a human subject having a cancer expressing CD33, including leukemia.
Method of treating cancer using CAR T cells
A method of treating cancer expressing CD33 in a subject by administering a therapeutically effective amount of a pharmaceutical composition containing T cells expressing the CAR, comprising the specified extracellular CD33 antigen binding domain, transmembrane domain, costimulatory and intracellular signaling domains as defined in the CAR structure.
Optional linker or spacer domain
The CAR's encoded CD33 antigen binding domain can be connected to the transmembrane domain by a linker or spacer domain derived from extracellular domains of proteins such as CD8, TNFRSF19, or CD28.
Intracellular CD3 zeta domain
The intracellular signaling domain of the CAR comprises a CD3 zeta intracellular domain.
Treatment of specific leukemia types
The leukemia treated includes acute myeloid leukemia (AML), blastic plasmacytoid dendritic cell neoplasm (BPDCN), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), acute lymphoblastic T cell leukemia (T-ALL), or acute lymphoblastic B cell leukemia (B-ALL).
The claims cover isolated nucleic acid molecules encoding CARs with fully human anti-CD33 binding domains linked to specific transmembrane, costimulatory, and intracellular signaling domains, vectors and cells comprising these nucleic acids, pharmaceutical compositions including CAR T cells, and methods of treating leukemia using these CAR T cells, providing a comprehensive protection for the novel CAR constructs and their therapeutic applications.
Stated Advantages
The CARs exhibit high surface expression on transduced T cells.
The CARs demonstrate a high degree of cytolytic activity against CD33-positive tumor cells.
The CAR T cells expand and persist in vivo at high levels after administration.
Using entirely human extracellular CD33 antigen binding domains reduces anti-CAR immune responses, enhancing function and persistence compared to mouse-derived binders.
Lower affinity binders allow tuning of efficacy versus toxicity and tumor specificity, potentially reducing off-target effects and bystander cell killing.
Documented Applications
Treatment or prevention of cancers expressing CD33, including acute myeloid leukemia (AML), blastic plasmacytoid dendritic cell neoplasm (BPDCN), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), acute lymphoblastic T cell leukemia (T-ALL), and acute lymphoblastic B cell leukemia (B-ALL).
Methods of making CAR T cells by transducing T cells with nucleic acid molecules encoding the anti-CD33 CARs.
Methods of diagnosing CD33-expressing diseases by contacting cells with human anti-CD33 antibodies or fragments and detecting CD33 expression.
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