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US-12624104-B2 - Compositions and methods for treating cancer with anti-CD19 immunotherapy

US12624104B2US 12624104 B2US12624104 B2US 12624104B2US-12624104-B2

Abstract

Chimeric antigen receptors containing human CD19 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.

Inventors

  • Dina Schneider
  • Rimas J. Orentas
  • Boro Dropulic
  • Dimiter S. Dimitrov
  • Zhongyu Zhu

Assignees

  • LENTIGEN TECHNOLOGY, INC.
  • THE U.S.A., AS REPRESENTED BY THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES

Dates

Publication Date
20260512
Application Date
20230607

Claims (20)

  1. 1 . A method of generating a population of RNA-engineered cells comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, where the RNA comprises an isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR) comprising at least one extracellular antigen binding domain comprising a CD19 antigen binding domain comprising the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, 14, or 16, at least one transmembrane domain, at least one intracellular signaling domain, and at least one linker or spacer domain.
  2. 2 . The method of claim 1 , wherein the at least one transmembrane domain comprises a transmembrane domain of a protein selected from an alpha chain of a T-cell receptor, a beta chain of the T-cell receptor, the zeta chain of the T-cell receptor, CD8, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, or any combination thereof.
  3. 3 . The method of claim 1 , wherein the at least one CD19 antigen binding domain, the at least one intracellular signaling domain, or both are connected to the transmembrane domain by the at least one linker or spacer domain.
  4. 4 . The method of claim 3 , wherein the at least one linker or spacer domain is from the extracellular domain of CD8 or CD28, and is linked to the at least one transmembrane domain.
  5. 5 . The method of claim 1 , wherein the nucleic acid sequence encoding the CD19 antigen binding domain comprises a nucleic sequence comprising SEQ ID NO. 5, 7, 9, 11, 13, or 15, or a sequence with 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereof.
  6. 6 . The method of claim 1 , wherein the at least one intracellular signaling domain comprises a signaling domain of CD3 zeta.
  7. 7 . The method of claim 1 , wherein the at least one intracellular signaling domain comprises a costimulatory domain, a primary signaling domain, or any combination thereof.
  8. 8 . The method of claim 7 , wherein the costimulatory domain comprises a functional signaling domain of OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), DAP10, DAP12, 4-1BB (CD137), or any combination thereof.
  9. 9 . A method of providing an anti-tumor immunity in a mammal comprising administering to the mammal an effective amount of an isolated cell comprising a vector comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR) comprising at least one extracellular antigen binding domain comprising a CD19 antigen binding domain comprising the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, 14, or 16, at least one transmembrane domain, at least one intracellular signaling domain, and at least one linker or spacer domain.
  10. 10 . The method of claim 9 , wherein the at least one transmembrane domain comprises a transmembrane domain of a protein selected from an alpha chain of a T-cell receptor, a beta chain of the T-cell receptor, the zeta chain of the T-cell receptor, CD8, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, or any combination thereof.
  11. 11 . The method of claim 9 , wherein the at least one CD19 antigen binding domain, the at least one intracellular signaling domain, or both are connected to the transmembrane domain by the at least one linker or spacer domain.
  12. 12 . The method of claim 11 , wherein the at least one linker or spacer domain is from the extracellular domain of CD8 or CD28, and is linked to the at least one transmembrane domain.
  13. 13 . The method of claim 9 , wherein the nucleic acid sequence encoding the CD19 antigen binding domain comprises a nucleic sequence comprising SEQ ID NO. 5, 7, 9, 11, 13, or 15, or a sequence with 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereof.
  14. 14 . The method of claim 9 , wherein the at least one intracellular signaling domain comprises a signaling domain of CD3 zeta.
  15. 15 . The method of claim 9 , wherein the at least one intracellular signaling domain comprises a costimulatory domain, a primary signaling domain, or any combination thereof.
  16. 16 . The method of claim 15 , wherein the costimulatory domain comprises a functional signaling domain of OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), DAP10, DAP12, 4-1BB (CD137), or any combination thereof.
  17. 17 . A method of treating cancer in a mammal, comprising administering to the mammal a CAR comprising at least one extracellular antigen binding domain comprising a CD19 antigen binding domain comprising the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, 14, or 16, at least one transmembrane domain, at least one intracellular signaling domain, and at least one linker or spacer domain, in an amount effective to treat cancer in the mammal.
  18. 18 . The method of claim 17 , wherein the at least one transmembrane domain comprises a transmembrane domain of a protein selected from an alpha chain of a T-cell receptor, a beta chain of the T-cell receptor, the zeta chain of the T-cell receptor, CD8, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, or any combination thereof.
  19. 19 . The method of claim 17 , wherein the at least one CD19 antigen binding domain, the at least one intracellular signaling domain, or both are connected to the transmembrane domain by the at least one linker or spacer domain.
  20. 20 . The method of claim 19 , wherein the at least one linker or spacer domain is from the extracellular domain of CD8 or CD28, and is linked to the at least one transmembrane domain.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a divisional application of U.S. patent application Ser. No. 16/707,505, filed on Dec. 9, 2019, which is a divisional application of U.S. patent application Ser. No. 16/132,064, issued as U.S. Pat. No. 10,501,539, which claims the benefit of priority under 35 U.S.C. Section 119(e) to U.S. Provisional Patent Application No. 62/559,297 filed on Sep. 15, 2017, the entire contents of which are incorporated herein by reference. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was created in the performance of a Cooperative Research and Development Agreement with the National Institutes of Health, an Agency of the Department of Health and Human Services. The Government of the United States has certain rights in this invention. SEQUENCE LISTING This application contains a Sequence Listing that has been submitted electronically as an XML file named “42449-0016003_SL_ST26.XML.” The XML file, created on Jun. 6, 2023, is 73,060 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety. FIELD OF THE DISCLOSURE This application relates to the field of cancer, particularly to CD19 antigen binding domains and chimeric antigen receptors (CARs) containing such CD19 antigen binding domains and methods of use thereof. BACKGROUND Cancer is one of the most deadly threats to human health. In the U.S. alone, cancer affects nearly 1.3 million new patients each year, and is the second leading cause of death after cardiovascular disease, accounting for approximately 1 in 4 deaths. Solid tumors are responsible for most of those deaths. Although there have been significant advances in the medical treatment of certain cancers, the overall 5-year survival rate for all cancers has improved only by about 10% in the past 20 years. Cancers, or malignant tumors, metastasize and grow rapidly in an uncontrolled manner, making treatment extremely difficult. CD19 is a 85-95 kDa transmembrane cell surface glycoprotein receptor. CD19 is a member of immunoglobulin (Ig) superfamily of proteins, and contains two extracellular Ig-like domains, a transmembrane, and an intracellular signaling domain (Tedder T F, Isaacs, C M, 1989, J Immunol 143:712-171). CD19 modifies B cell receptor signaling, lowering the triggering threshold for the B cell receptor for antigen (Carter, RH, and Fearon, DT, 1992, Science, 256:105-107), and co-ordinates with CD81 and CD21 to regulate this essential B cell signaling complex (Bradbury, L E, Kansas G S, Levy S, Evans R L, Tedder T F, 1992, J Immunol, 149:2841-50). During B cell ontogeny CD19 is able to signal at the pro-B, pre-pre-B cell, pre-B, early B cell stages independent of antigen receptor, and is associated with Src family protein tyrosine kinases, is tyrosine phosphorylated, inducing both intracellular calcium mobilization and inositol phospholipid signaling (Uckun F M, Burkhardt A L, Jarvis L, Jun X, Stealy B, Dibirdik I, Myers D E, Tuel-Ahlgren L, Bolen J B, 1983, J Biol Chem 268:21172-84). The key point of relevance for treatment of B cell malignancies is that CD19 is expressed in a tightly regulated manner on normal B cells, being restricted to early B cell precursors at the stage of IgH gene rearrangement, mature B cells, but not expressed on hematopoietic stem cells, or mature plasma cells (Anderson, K C, Bates, M P, Slaughenhout B L, Pinkus G S, Schlossman S F, Nadler L M, 1984, Blood 63:1424-1433). The present standard of care for B-lineage leukemias may consists of remission induction treatment by high dose of chemotherapy or radiation, followed by consolidation, and may feature stem cell transplantation and additional courses of chemotherapy as needed (see the world wide web at cancer.gov). High toxicity associated with these treatments, as well as the risk of complications, such as relapse, secondary malignancy, or GVHD, motivate the search for better therapeutic alternatives. The expression of CD19 on both adult and pediatric (pre-B-ALL) B cell malignancies has led to exploiting this target for both antibody and chimeric antigen receptor (CAR)-T cell-based therapy (Kochenderfer J N, Wilson W H, Janik J E, Dudley M E, Stetler-Stevenson M, Feldman S A, Maric I, Raffeld M, Nathan D A, Lanier B J, Morgan R A, Rosenberg S A, 2010, Blood 116:4099-102; Lee D W, Kochenderfer J N, Stetler-Stevenson M, Cui Y K, Delbrook C, Feldman S A, Orentas R, Sabatino M, Shah N N, Steinberg S M, Stroncek D, Tschemia N, Yuan C, Zhang H, Zhang L, Rosenberg S A, Wayne A S, Mackall C L, 2015, Lancet 385:517-28). A number of novel approaches to treat B cell leukemia and lymphoma have been developed, including bi-specific antibodies that link an anti-CD19 binding motif to a T cell binding motif (i.e. Blinatumomab, Blincyto® indicated for the treatment of Philadelphia chromosome-negative relapsed or refractory B-cell precursor acute lymphoblastic leukemia (ALL). To date, many of the binding moieties for CD