Modified chimeric receptors and related compositions and methods

Patent metadata

Field	Value
Publication number	US-2018355014-A1
Application number	US-201615780623-A
Country	US
Kind code	A1
Filing date	Dec 2, 2016
Priority date	Dec 3, 2015
Publication date	Dec 13, 2018
Grant date	—

Abstract

Provided are chimeric receptors for engineering cells for adoptive therapy, including T cells, and the genetically engineered cells. In some embodiments, the chimeric receptors, such as chimeric antigen receptors (CARs) are modified in a junction region by one or more amino acid modifications such that peptide fragments of such region exhibit a lower binding affinity for a human leukocyte antigen (HLA) and/or the region exhibits reduced immunogenicity, including following administration to a subject. In some aspects, also provided are methods and compositions for engineering and producing cells expressing such chimeric receptors, compositions containing the cells, and method for their administration to subjects. In some embodiments, features of the chimeric receptors and engineered cells containing the chimeric receptors result in methods that provide for increased or improved activity, efficacy and/or persistence.

First claim

What is claimed:

1 . A variant chimeric receptor, comprising a modified junction region having one or more amino acid sequence modifications compared to a junction region of a reference chimeric receptor, wherein:
the reference chimeric receptor comprises a first domain and a second domain, joined in contiguous sequence at a junction, wherein the junction region of the reference chimeric receptor comprises up to 15 contiguous amino acids directly C-terminal of the junction and/or up to 15 contiguous amino acids directly N-terminal of the junction; and a peptide fragment having the sequence of an 8-15 amino acid portion of the modified junction region has a binding affinity for a human leukocyte antigen (HLA) molecule that is lower than the binding affinity, for the same HLA molecule, of a peptide fragment having the sequence of the corresponding portion of the junction region of the reference chimeric receptor.

2 . The variant chimeric receptor of claim 1 , wherein the peptide fragment of the corresponding portion of the junction region of the reference chimeric receptor has a binding affinity of less than 1000 nM, less than 500 nM or less than 50 nM.

3 . A variant chimeric receptor, comprising a modified junction region having one or more amino acid sequence modifications compared to a junction region of a reference chimeric receptor, wherein:
the reference chimeric receptor comprises a first domain and a second domain, joined in contiguous sequence at a junction, wherein the junction region of the reference chimeric receptor comprises up to 15 contiguous amino acids directly C-terminal of the junction and/or up to 15 contiguous amino acids directly N-terminal of the junction; and the average of the binding affinities of all 8-15 amino acid fragments, or of all 8, 9, 10, 11, 12, 13, 14, or 15 amino acid fragments, within the modified junction region for a human HLA molecule is lower than the average of the binding affinities of all 8-15 amino acid fragments, or of all 8, 9, 10, 11, 12, 13, 14, or 15 amino acid fragments, within the junction region of the reference chimeric receptor.

4 . The variant chimeric receptor of any of claims 1 - 3 , wherein the binding affinity or average of binding affinities is more than 2-fold, more than 5-fold, more than 10-fold, more than 25-fold, more than 50-fold or more than 100-fold lower.

5 . A variant chimeric receptor, comprising a modified junction region having one or more amino acid sequence modifications compared to a junction region of a reference chimeric receptor, wherein:
the reference chimeric receptor comprises a first domain directly linked to a second domain joined in contiguous sequence at a junction, wherein; the junction region of the reference chimeric receptor comprises up to 15 contiguous amino acids directly C-terminal of the junction and/or up to 15 contiguous amino acids directly N-terminal of the junction; and the number of peptide fragments having the sequence of an 8-15 amino acid portion of the modified junction region that has a binding affinity for a human leukocyte antigen (HLA) of less than 1000 nM is reduced compared to the number of peptide fragments having the sequence of an 8-15 amino acid portion of the junction region of the reference chimeric receptor that has the same affinity for binding the same HLA.

6 . The variant chimeric receptor of claim 5 , wherein:
the number of peptide fragments within the modified junction region that exhibits a binding affinity for an HLA of less than 500 nM or less than 50 nM is reduced; or the binding affinity of less than 1000 nM is a binding affinity of less than 500 nM or less than 50 nm.

7 . The variant chimeric receptor of any of claims 1 - 6 , wherein the binding affinity is an IC50 and the comparison of binding of peptide fragments of the modified junction regions and the peptide fragments of the junction region of the reference chimeric receptor is with reference to the same standard peptide.

8 . The variant chimeric receptor of any of claims 1 - 7 , wherein:
the first domain and/or second domain comprises a domain of a natural human protein; and/or the first domain and/or second domain comprises an extracellular binding domain, a hinge domain, a transmembrane domain, or an intracellular signaling domain, which intracellular signaling domain is, optionally, a costimulatory signaling domain or an activating cytoplasmic signaling domain.

9 . The variant chimeric receptor of any of claims 1 - 8 , wherein the first domain and second domain are not present in the same molecule in vivo in a human subject.

10 . The variant chimeric receptor of any of claims 1 - 9 , wherein the first domain and second domain are, respectively, an extracellular ligand binding domain and a hinge domain, a hinge domain and a transmembrane domain, a transmembrane domain and an intracellular costimulatory signaling domain, and an intracellular costimulatory signaling domain and an activating cytoplasmic signaling domain, which can include functional portions of such domains.

11 . The variant chimeric receptor of any of claims 1 - 10 , wherein the first domain is a transmembrane domain or a functional portion thereof and the second domain is a costimulatory signaling domain or a functional portion thereof.

12 . The variant chimeric receptor of claim 11 , wherein the transmembrane domain is a CD28 transmembrane domain or a functional portion or variant thereof and the costimulatory signaling domain is a 4-1BB signaling domain or a functional portion or variant thereof.

13 . The variant chimeric receptor of any of claims 1 - 12 , wherein the junction region of the reference chimeric receptor comprises up to 13 contiguous amino acids directly C-terminal of the junction and/or up to 15 contiguous amino acids directly N-terminal of the junction.

14 . The variant chimeric receptor of any of claims 1 - 13 , wherein the peptide fragment(s) comprises a sequence of amino acids between or between about 8 and 15 amino acids in length, or comprises a sequence of amino acids that is at least or at least about or is or is about 8, 9, 10, 11, 12, 13, 14 or 15 amino acids in length.

15 . The variant chimeric receptor of any of claims 1 - 14 , wherein the variant chimeric receptor comprises:
a domain of at least 95% sequence identity to the first domain and/or a domain of at least 95% sequence identity to the second domain; a domain identical in sequence to the first domain and a domain of at least 95% sequence identity to the second domain; or a domain of at least 95% sequence identity to the first domain and a domain identical in sequence to the second domain, wherein at least one or both of the domains present in the variant chimeric receptor is modified compared to the first domain and/or second domain of the reference chimeric receptor in the portion comprising the modified junction region.

16 . The variant chimeric receptor of any of claims 1 - 15 , wherein:
the variant chimeric receptor comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to the reference chimeric receptor; and/or the variant chimeric receptor comprises up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acid modifications compared to the reference chimeric receptor.

17 . The variant chimeric receptor of any of claims 12 - 16 , wherein
the CD28 transmembrane domain comprises the sequence of amino acids set forth in SEQ ID NO:2, 103 or 104 or a functional portion or variant thereof comprising a seque

Assignees

• Juno Therapeutics Inc

Inventors

• Thompson Lucas James
• Dubose Robert F
• Brahmandam Archana

Classifications

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• C07K14/70539 Primary

  MHC-molecules, e.g. HLA-molecules · CPC title

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title

• C07K16/2815

  against CD8 · CPC title