Targeted protein degradation to attenuate adoptive T-cell therapy associated adverse inflammatory responses

Patent metadata

Field	Value
Publication number	US-11046954-B2
Application number	US-201815889963-A
Country	US
Kind code	B2
Filing date	Feb 6, 2018
Priority date	Aug 6, 2015
Publication date	Jun 29, 2021
Grant date	Jun 29, 2021

Abstract

This invention is in the area of compositions and methods for regulating chimeric antigen receptor immune effector cell, for example T-cell (CAR-T), therapy to modulate associated adverse inflammatory responses, for example, cytokine release syndrome and tumor lysis syndrome, using targeted protein degradation.

First claim

We claim:

1. A therapeutic system for degrading a chimeric antigen receptor expressed in a T-cell, the system comprising:
a. a T-cell comprising a chimeric antigen receptor polypeptide, wherein the chimeric antigen receptor polypeptide comprises:
i) an extracellular ligand binding protein that binds CD19 and has the amino acid sequence of SEQ ID NO: 10, or wherein the extracellular ligand binding protein binds Erb2 and has an amino acid sequence comprising the amino acid sequence of SEQ ID NO: 20 and the amino acid sequence of SEQ ID NO: 21, or wherein the extracellular ligand binding protein binds Erb2 and has an amino acid sequence comprising the amino acid sequence of SEQ ID NO: 20 and the amino acid sequence of SEQ ID NO: 22;
ii) a hinge region which has the amino acid sequence of SEQ ID NO: 15 and a transmembrane protein which has the amino acid sequence of SEQ ID NO: 16;
iii) a cytoplasmic protein comprising at least one intracellular signaling protein which has the amino acid sequence of SEQ ID NO: 17; and,
iv) a heterobifunctional compound targeting protein capable of being bound by a heterobifunctional compound, wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 9; and

b. a heterobifunctional compound capable of binding to i) the chimeric antigen receptor polypeptide through the heterobifunctional compound targeting protein and ii) a ubiquitin ligase;
wherein the chimeric antigen receptor polypeptide, when bound by the heterobifunctional compound, is capable of being ubiquitinated and then degraded by a proteasome, provided that when the heterobifunctional targeting protein is SEQ ID NO: 1, the heterobifunctional compound is selected from:

and when the heterobifunctional targeting protein is SEQ ID NO: 2, the heterobifunctional compound is selected from:

and when the heterobifunctional targeting protein is SEQ ID NO: 3, the heterobifunctional compound is selected from:

and when the heterobifunctional targeting protein is SEQ ID NO: 9, the heterobifunctional compound is selected from:

2. The therapeutic system of claim 1 , wherein the T-cell is an autologous human T-cell.

3. The therapeutic system of claim 1 , wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 1 and wherein the heterobifunctional compound is selected from:

4. The therapeutic system of claim 1 , wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 2 and wherein the heterobifunctional compound is selected from:

5. The therapeutic system of claim 1 , wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 3 and wherein the heterobifunctional compound is selected from:

6. The therapeutic system of claim 1 , wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 9 and wherein the heterobifunctional compound is selected from:

7. A T-cell comprising a chimeric antigen receptor polypeptide, wherein the chimeric antigen receptor polypeptide comprises:
i) an extracellular ligand binding protein that binds CD19 and has the amino acid sequence of SEQ ID NO: 10, or wherein the extracellular ligand binding protein binds Erb2 and has an amino acid sequence comprising the amino acid sequence of SEQ ID NO: 20 and the amino acid sequence of SEQ ID NO: 21, or wherein the extracellular ligand binding protein binds Erb2 and has an amino acid sequence comprising the amino acid sequence of SEQ ID NO: 20 and the amino acid sequence of SEQ ID NO: 22;
ii) a hinge region which has the amino acid sequence of SEQ ID NO: 15 and a transmembrane protein which has the amino acid sequence of SEQ ID NO: 16;
iii) a cytoplasmic protein comprising at least one intracellular signaling protein which has the amino acid sequence of SEQ ID NO: 17; and,
iv) a heterobifunctional compound targeting protein capable of being bound by a heterobifunctional compound;
wherein the heterobifunctional compound targeting protein comprises the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 9;
wherein the heterobifunctional compound is capable of binding to i) the chimeric antigen receptor polypeptide through the heterobifunctional compound targeting protein and ii) a ubiquitin ligase;
wherein the chimeric antigen receptor polypeptide, when bound by the heterobifunctional compound, is capable of being ubiquitinated and degraded by a proteasome,
provided that when the heterobifunctional targeting protein is SEQ ID NO: 1, the heterobifunctional compound is selected from:

and when the heterobifunctional targeting protein is SEQ ID NO: 2, the heterobifunctional compound is selected from:

and when the heterobifunctional targeting protein is SEQ ID NO: 3, the heterobifunctional compound is selected from:

and when the heterobifunctional targeting protein is SEQ ID NO: 9, the heterobifunctional compound is sele

Assignees

• Dana Farber Cancer Inst Inc

Inventors

• Bradner James
• Roberts Justin
• Nabet Behnam
• Winter Georg
• Phillips Andrew J
• Heffernan Timothy
• Buckley Dennis

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

• C07K16/00 Primary

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title