Methods for Enhancing Efficacy of Therapeutic Immune Cells

Patent metadata

Field	Value
Publication number	US-2018008638-A1
Application number	US-201615548577-A
Country	US
Kind code	A1
Filing date	Feb 5, 2016
Priority date	Feb 6, 2015
Publication date	Jan 11, 2018
Grant date	—

Abstract

The present invention relates to a method of using a receptor (e.g., chimeric antigen receptor—CAR) that activates an immune response upon binding a cancer cell ligand in conjunction with a target-binding molecule that targets a protein or molecule CI for removal or neutralization to generate enhanced anti-cancer immune cells. The present invention also relates to engineered immune cells having enhanced therapeutic efficacy and uses thereof.

First claim

What is claimed is:

1 . An engineered immune cell that comprises a nucleic acid comprising a nucleotide sequence encoding an immune activating receptor and a nucleic acid comprising a nucleotide sequence encoding a target-binding molecule linked to a localizing domain.

2 . The engineered immune cell of claim 1 , wherein the engineered immune cell is an engineered T cell, an engineered natural killer (NK) cell, an engineered NK/T cell, an engineered monocyte, an engineered macrophage, or an engineered dendritic cell.

3 . The engineered immune cell of claim 1 , wherein the receptor is a chimeric antigen receptor (CAR).

4 . The engineered immune cell of claim 1 , wherein the CAR is anti-CD19-4-1BB-CD3ζ CAR.

5 . The engineered immune cell of claim 1 , wherein the target-binding molecule is an antibody.

6 . The engineered immune cell of claim 5 , wherein the antibody is a single-chain variable fragment (scFv).

7 . The engineered immune cell of claim 5 , wherein the antibody binds to a factor in a CD3/T-cell receptor (TCR) complex, a cytokine, a human leukocyte antigen (HLA) Class I molecule, or a receptor that downregulates immune response.

8 . The engineered immune cell of claim 7 , wherein the factor in a CD3/TCR complex is CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, or CD3ζ.

9 . The engineered immune cell of claim 7 , wherein the HLA Class I molecule is β2-microglobulin, α1-microglobulin, α2-microglobulin, or α3-microglobulin.

10 . The engineered immune cell of claim 7 , wherein the receptor that downregulates immune response is selected from programmed cell death protein 1 (PD-1), cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), T-cell immunoglobulin and mucin-domain containing-3 (Tim3), killer immunoglobulin-like receptors (KIRs), CD94, NKG2A, or a protein tyrosine phosphatase.

11 . The engineered immune cell of claim 7 , wherein the cytokine is interleukin (IL)-6, IL-2, IL-4, IL-7, IL-10, IL-12, IL-15, IL-18, IL-21, IL-27, IL-35, interferon (IFN)-γ, IFN-β, IFN-α, tumor necrosis factor (TNF)-α, or transforming growth factor (TGF)-β.

12 . The engineered immune cell of claim 1 , wherein the target-binding molecule binds to CD2, CD4, CD5, CD7, CD8, CD30, CD38, CD45, CD52, or CD127.

13 . The engineered immune cell of claim 7 , wherein the localizing domain comprises an endoplasmic reticulum (ER) or Golgi retention sequence; a proteosome localizing sequence; a transmembrane domain sequence derived from CD8α, CD8β, 4-1BB, CD28, CD34, CD4, FcεRIγ, CD16, OX40, CD3ζ, CD3ε, CD3γ, CD3δ, TCRα, CD32, CD64, VEGFR2, FAS, or FGFR2B.

14 . The engineered immune cell of claim 13 , wherein the ER or Golgi retention sequence comprises the amino acid sequence KDEL (SEQ ID NO: 4), KKXX (SEQ ID NO: 9), KXD/E (SEQ ID NO: 10), or YQRL (SEQ ID NO: 11), wherein X is any amino acid.

15 . The engineered immune cell of claim 13 , wherein the proteasome localizing sequence comprises a PEST motif.

16 . The engineered immune cell of claim 1 , wherein the immune activating receptor is a non-naturally occurring molecule.

17 . The engineered immune cell of claim 1 , wherein the target-binding molecule linked to a localizing domain is a non-naturally occurring molecule.

18 . Use of the engineered immune cell of claim 1 for treating cancer, comprising administering a therapeutic amount of the engineered immune cell to a subject in need thereof.

19 . The use of claim 18 , wherein the engineered immune cell is administered into the subject by intravenous infusion, intra-arterial infusion, direct injection into tumor and/or perfusion of tumor bed after surgery, implantation at a tumor site in an artificial scaffold, intrathecal administration, and intraocular administration.

20 . The use of claim 18 , wherein the cancer is a solid tumor or a hematologic malignancy.

21 . A method for producing the engineered immune cell of claim 1 , the method comprising:
introducing into an immune cell a nucleic acid comprising a nucleotide sequence encoding an immune activating receptor and a nucleic acid comprising a nucleotide sequence encoding a target-binding molecule linked to a localizing domain, thereby producing an engineered immune cell.

22 . The method of claim 21 , wherein the engineered immune cell is produced ex vivo.

Assignees

• Nat Univ Singapore

Inventors

• Campana Dario
• Kamiya Takahiro

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P37/02

  Immunomodulators · CPC title

• C07K2319/06

  containing a lysosomal/endosomal localisation signal · CPC title

• C07K2319/05

  containing a GOLGI retention signal · CPC title

• C07K2319/04

  containing an ER retention signal such as a C-terminal HDEL motif · CPC title