Lentiviral vectors for regulated expression of a chimeric antigen receptor molecule

Patent metadata

Field	Value
Publication number	US-10752668-B2
Application number	US-201515327517-A
Country	US
Kind code	B2
Filing date	Jul 24, 2015
Priority date	Jul 25, 2014
Publication date	Aug 25, 2020
Grant date	Aug 25, 2020

Abstract

The invention relates to the regulated expression of a chimeric antigen receptor (CAR) within a lentiviral vector. The CAR comprises a hook-binding domain that interacts with a hook, preferably encoded by the same lentiviral vector, which prevents proper processing and release of the CAR to the cell membrane. The invention encompasses vectors, methods of making the vectors, and methods of using them, including medicinal uses. The vectors can be used for administration to humans to induce immune responses and to treat cancers and tumors.

First claim

We claim:

1. A chimeric antigen receptor comprising:
a binding domain;
a transmembrane domain;
a hook-binding domain comprising a streptavidin-binding peptide; and
an activation domain comprising a T cell activating fragment comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence selected from the group consisting of:
the amino acid fragment of sequence PKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYD VLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKG HDGLY from SEQ ID NO:3;
SEQ ID NO:4;
the amino acid fragment of sequence KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL from SEQ ID NO:5;
SEQ ID NO:6;
SEQ ID NO:7,
SEQ ID NO:8,
SEQ ID NO:33, and
SEQ ID NO:34,
the hook-binding domain being encoded by the nucleic acid sequence SEQ ID NO:2;
the hook-binding domain being located in the intracytoplasmic region, between the transmembrane domain and the activation domain, between two activation domains or at the intracytoplasmic C-terminus of the chimeric antigen receptor;
the binding domain being located in the extracytoplasmic region of the chimeric antigen receptor; and
the T cell activating fragment being located in the intracytoplasmic region of the chimeric antigen receptor.

2. The chimeric antigen receptor of claim 1 , wherein the binding domain comprises a single-chain Fv antibody or a nanobody.

3. The chimeric antigen receptor of claim 1 , wherein the chimeric antigen receptor comprises the amino acid sequence of any of SEQ ID NO:46, SEQ ID NO:48, or SEQ ID NO:50.

4. The chimeric antigen receptor of claim 1 , wherein the activation domain comprises a T cell activating fragment comprising the amino acid sequence selected from the group consisting of:
the amino acid fragment of sequence PKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRG KGHDGLY from SEQ ID NO:3;
SEQ ID NO:4;
the amino acid fragment of sequence KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL from SEQ ID NO:5;
SEQ ID NO:6,
SEQ ID NO:7,
SEQ ID NO:8,
SEQ ID NO:33, and
SEQ ID NO:34.

5. The chimeric antigen receptor of claim 1 , wherein the activation domain comprises the amino acid fragment of sequence KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL from the amino acid sequence SEQ ID NO:5 and
the amino acid fragment of sequence PKLCYLLD GILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRG KGHDGLY from the amino acid sequence SEQ ID NO:3.

Assignees

• Theravectys
• Inst Curie
• Centre Nat Rech Scient
• Univ Paris Curie
• Institut National De La Sante Et De La Rech Medicale (Inserm)
• Univ Sorbonne

Inventors

• Agaugue Sophie
• Tibaldi Lorenzo
• Even-Desrumaux Klervi
• Trubetskoy Dmitry
• Perez Franck
• Amigorena Sebastian
• Bauche Cécile

Classifications

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/4205

  Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4 · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/23

  On/off switch · CPC title