High affinity monoclonal antibodies targeting glypican-1 and methods of use
Inventors
Ho, Mitchell • Li, Nan • PAN, JIAJIA
Assignees
US Department of Health and Human Services
Publication Number
US-12122843-B2
Publication Date
2024-10-22
Expiration Date
2040-01-15
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Abstract
Monoclonal antibodies that specifically bind glypican-1 (GPC1) are described. Chimeric antigen receptor (CAR) T cells, immunotoxins and other antibody conjugates based on the GPC1-specific antibodies are also described. The disclosed CAR T cells, immunotoxins, GPC1-specific antibodies and conjugates thereof can be used, for example, in the diagnosis or treatment of GPC1-positive pancreatic cancer and other cancers.
Core Innovation
The invention describes monoclonal antibodies that specifically bind glypican-1 (GPC1) with high affinity, such as the mouse monoclonal antibody HM2 and the camel single-domain monoclonal antibody D4. These antibodies can be used to generate chimeric antigen receptor (CAR) T cells, immunotoxins, antibody-drug conjugates (ADCs), multi-specific antibodies, antibody-nanoparticle conjugates, and fusion proteins. The CAR T cells comprising these antibodies are capable of potently killing GPC1-positive tumor cells both in vitro and in vivo.
The problem addressed is the urgent need for effective therapies targeting solid tumors, including pancreatic cancer, which has a low survival rate and limited effective treatments. GPC1 is overexpressed in various cancers such as pancreatic, breast, glioma, colorectal, and ovarian cancers. While CAR T cell immunotherapy has shown promise in hematological malignancies, there remains a need to identify and validate new CAR T cell targets for solid tumors like pancreatic cancer.
To solve this, the invention provides monoclonal antibodies that specifically bind GPC1 with high affinity, enabling the creation of CAR T cells and other antibody conjugates directed against GPC1-positive cancers. These antibodies and conjugates also can be used for diagnostic purposes, for example, detecting GPC1 expression in biological samples. Moreover, the invention includes nucleic acid molecules and vectors for producing the antibodies and CAR constructs, as well as isolated cells expressing GPC1-specific CARs.
Claims Coverage
The patent includes 39 claims with independent claims focused on monoclonal antibodies specific for GPC1, CARs incorporating these antibodies, isolated cells expressing the CARs, immunoconjugates, ADCs, multi-specific antibodies, antibody-nanoparticle conjugates, fusion proteins, nucleic acid encoding the antibodies and CARs, and therapeutic and diagnostic methods using these components.
Monoclonal antibodies with specific CDR sequences binding GPC1
The monoclonal antibody specifically binds GPC1 and comprises either a VH domain and a VL domain containing CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 2 and SEQ ID NO: 4 respectively, or is a single-domain antibody comprising CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 6.
Chimeric antigen receptors comprising GPC1-specific antibodies
CARs comprising the monoclonal antibody that binds GPC1, optionally including a hinge region (such as a CD8α hinge), transmembrane domain (such as CD8α transmembrane), costimulatory signaling moiety (such as 4-1BB), and signaling domain (such as CD3ζ).
Isolated cells expressing the GPC1-targeted CARs
Isolated cells, including cytotoxic T lymphocytes (CTLs) or natural killer (NK) cells, expressing the disclosed CARs incorporating the GPC1-specific monoclonal antibodies.
Immunoconjugates and immunotoxins incorporating GPC1 antibodies
Immunoconjugates comprising the GPC1-specific monoclonal antibody and an effector molecule such as a toxin, including Pseudomonas exotoxin variants like PE-LR, with amino acid sequences corresponding to SEQ ID NOs: 15, 17, 19, or 21.
Antibody-drug conjugates with cytotoxic agents attached to GPC1 antibodies
ADCs comprising a drug conjugated to the GPC1-specific monoclonal antibody, wherein the drug may be a cytotoxic agent like anti-microtubule or cross-linking agents.
Multi-specific antibodies including the GPC1-specific antibody
Multi-specific antibodies containing the GPC1-specific monoclonal antibody and at least one additional monoclonal antibody or antigen-binding fragment thereof.
Antibody-nanoparticle conjugates
Conjugates comprising a nanoparticle linked to the GPC1-specific monoclonal antibody for targeted delivery.
Fusion proteins containing the GPC1-specific monoclonal antibody
Fusion proteins comprising the GPC1-specific monoclonal antibody and a heterologous protein or peptide.
Nucleic acid molecules and vectors encoding the antibodies and CARs
Isolated nucleic acids encoding the disclosed monoclonal antibodies, immunoconjugates, and CARs, including compositions with promoters, and vectors comprising these nucleic acids.
Therapeutic and diagnostic methods
Methods of treating GPC1-positive cancers by administering the disclosed antibodies, CARs, immunoconjugates, ADCs or compositions thereof; and methods of detecting GPC1 expression or diagnosing GPC1-positive cancers by contacting a sample with the monoclonal antibody and detecting binding.
The claims collectively cover monoclonal antibodies specifically binding GPC1 with defined CDR sequences, CARs and cells expressing them, immunoconjugates including immunotoxins with Pseudomonas exotoxin variants, ADCs, multi-specific antibodies, nanoparticle conjugates, fusion proteins, nucleic acid constructs for production, and therapeutic and diagnostic applications targeting GPC1-positive cancers.
Stated Advantages
The disclosed GPC1-specific monoclonal antibodies bind with high affinity and specificity to GPC1.
CAR T cells generated using the disclosed antibodies show potent cytolytic activity against GPC1-positive tumor cells in vitro and in vivo.
Immunotoxins based on the disclosed antibodies retain binding affinity and selectively kill GPC1-expressing cells while sparing GPC1-negative cells.
Bivalent immunotoxins enhance binding and cytotoxicity, particularly in native pancreatic cancer cells with lower GPC1 expression.
The compositions can be used for diagnosis and treatment, providing effective targeting of GPC1-positive cancers with potential for reduced off-target effects.
Documented Applications
Diagnosis and treatment of GPC1-positive cancers, including pancreatic cancer, colorectal cancer, liver cancer, glioma, lung cancer, head and neck cancer, thyroid cancer, endometrial cancer, breast cancer, and ovarian cancer.
Engineering CAR T cells and NK cells that target GPC1-positive tumor cells.
Use of immunotoxins, antibody-drug conjugates, multispecific antibodies, and antibody-nanoparticle conjugates for targeted cancer therapy.
Detection of GPC1 expression in biological samples, including serum samples containing exosomes, for cancer diagnosis or confirmation.
Use of labeled antibodies for in vivo tumor imaging and diagnostic methods including immunoPET.
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