Chimeric antigen receptor (CAR) modulation
Inventors
Ngo, John • Wong, Wilson • WONG, Meng Lai Nicole • LI, Huishan • TAGUE, Elliot P.
Assignees
Publication Number
US-12202863-B2
Publication Date
2025-01-21
Expiration Date
2040-03-27
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Abstract
The technology described herein is directed to CAR polypeptides and systems comprising repressible proteases. In combination with a specific protease inhibitor, the activity of said CAR polypeptides and systems and cells comprising them can be modulated. Also described herein are methods of using said CAR polypeptides and systems, for example to treat various diseases and disorders.
Core Innovation
The invention provides drug-inducible chimeric antigen receptor (CAR) polypeptide systems comprising a reader domain and a repressible protease, allowing precise modulation of CAR activity. The core of the system consists of a first polypeptide that includes an extracellular binding domain, a transmembrane domain, and a reader domain (either a danoprevir/NS3 complex reader domain (DNCR) or a grazoprevir/NS3 reader complex (GNCR) domain), and a second polypeptide that includes a repressible protease based on hepatitis C virus (HCV) nonstructural protein 3 (NS3), specifically genotype 1B, and at least one intracellular signaling domain.
The problem addressed is the lack of controllable safety-switch technologies in existing CAR-T therapies, resulting in issues such as T cell overactivity and potentially severe adverse effects like cytokine release syndrome. Existing inducible CAR systems either use protein-based inducers that are difficult to administer, small molecules with poor pharmacokinetics, or non-FDA-approved molecules. There is a critical need for CAR technologies that can be precisely regulated using clinically approved drugs with favorable toxicity and pharmacokinetic profiles.
This invention solves that problem by integrating a drug-responsive, small molecule-controlled mechanism into CAR systems. The reader domain in the first polypeptide specifically binds the NS3 protease of the second polypeptide only in the presence of a specific protease inhibitor (such as grazoprevir or danoprevir). This enables the activation or inactivation of the CAR system to be controlled in a reversible and precise manner with the administration or withdrawal of an FDA-approved protease inhibitor, providing flexible and customizable control over CAR activity in immune cells.
Claims Coverage
The claims define a polypeptide system comprising a reader-based CAR construct and a second polypeptide containing an HCV NS3 protease domain with precise inventive features covering their composition, binding specificity, and activity modulation.
Drug-inducible reader CAR system with specific binding
The system consists of: - A first polypeptide comprising an extracellular binding domain, a transmembrane domain, and a reader domain that is either a DNCR or GNCR domain. - A second polypeptide comprising a repressible protease (HCV NS3, genotype 1B; SEQ ID NO: 99, 130, or 180) and at least one intracellular signaling domain. - The reader domain specifically binds the repressible protease in the presence of a corresponding protease inhibitor (e.g., grazoprevir or danoprevir).
Protease and reader domain configuration and catalytic inactivity
The claims encompass: - The repressible protease (NS3) may be catalytically dead. - The first polypeptide may not contain a protease cleavage site. - The second polypeptide may not contain a protease cleavage site. - The repressible protease of the second polypeptide can be positioned between various domains (e.g., between the transmembrane domain and the at least one intracellular signaling domain; between the first and second intracellular signaling domains; or at the C terminus).
Combinations and sequence variants of polypeptide constructs
The claims further cover: - Specific sequence configurations, such as polypeptides comprising residues 1-626 of SEQ ID NO: 127, residues 1-630 of SEQ ID NO: 128, SEQ ID NOs: 86-89, residues 648-1303 of SEQ ID NO: 127, or residues 652-1307 of SEQ ID NO: 128, or sequences at least 70% identical thereto that maintain the same function. - First and second polypeptides that are physically linked or flank a self-cleaving peptide domain.
Use of FDA-approved protease inhibitors for system activation
The system is claimed as being in combination with a protease inhibitor bound to the repressible protease, where the inhibitor is selected from FDA-approved drugs such as grazoprevir, danoprevir, simeprevir, asunaprevir, ciluprevir, boceprevir, sovaprevir, paritaprevir, ombitasvir, ritonavir, dasabuvir, or telaprevir. The activity of the reader CAR system is dictated by the presence of the protease inhibitor that corresponds to the reader domain.
Inclusion of signal and cofactor domains in polypeptides
The claims optionally specify: - The presence of at least one intracellular signaling domain selected from a defined set including CD3zeta, CD28, 4-1BB, and others. - Inclusion of an HSV NS4A domain as a cofactor for the repressible protease in the second polypeptide.
In summary, the patent claims cover a drug-inducible, reader-based CAR polypeptide system utilizing a specific binding interaction between engineered reader domains and HCV NS3 protease (genotype 1B), modulated by FDA-approved protease inhibitors, with claim scope including various construct permutations, specific sequence variants, and cofactor configurations.
Stated Advantages
The CAR system provides precise, reversible, and customizable control over CAR activity using FDA-approved small molecule drugs.
The invention improves the safety of adoptive cell therapies by enabling drug-mediated modulation of CAR activity, reducing risks such as cytokine release syndrome and other toxicities.
The system allows for flexible manipulation of immune cell function by controlling the assembly and signaling of CAR constructs in response to clinically-relevant drugs.
Unlike prior inducible CAR technologies, this invention uses readily-administered, clinically-approved drugs with known safety and pharmacokinetic profiles.
Documented Applications
Modulation of adoptive cell therapies, including CAR-T, CAR-NK, and CAR-Treg cell therapies, for treatment of cancer by controlling immune cell activation using small molecule protease inhibitors.
Provision of safety control switches for immune cell-based therapies, enabling clinicians to modulate or halt CAR activity in response to adverse events such as cytokine release syndrome.
Generation of cell-based therapies with adjustable activity levels in response to the administration or withdrawal of protease inhibitors, for increased therapeutic precision in oncology.
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