Chimeric proteins and methods of immunotherapy

• US 10,336,807 B2
• Filed: 07/06/2018
• Issued: 07/02/2019
• Est. Priority Date: 01/11/2016
• Status: Active Grant

• Current Assignee: Rufuge Biotechnologies, Inc., Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Inventors: Qi, Lei S., Wang, Bing C
• Primary Examiner(s): Goddard, Laura B
• Application Number:
  US16/029,299
• Publication Number:
  US 20180346543A1
• Time in Patent Office: 361 Days
• Field of Search: None
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/4636 Immune checkpoint inhibitors
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/01 containing a localisation/targetting motif
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• CPC Class Codes: C07K 2319/09 containing a nuclear localisation signal
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/74 containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C12N 15/1086 Preparation or screening of expression libraries, e.g. reporter assays
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present disclosure provides systems for immune cell regulation and methods of immunotherapy. Systems of the present disclosure for immune cell regulation comprise a chimeric receptor polypeptide, a chimeric adaptor polypeptide, a gene modulating polypeptide (GMP), and a cleavage moiety.

Claims

1. A method of inducing death of a target cell, comprising:

(a) expressing a system in a lymphocyte; and

(b) contacting said target cell with the lymphocyte under conditions that induce said death of the target cell,

wherein the system expressed in the lymphocyte comprises;

(i) a chimeric transmembrane receptor polypeptide (receptor) comprising a ligand binding domain, an immune cell signaling domain, and a gene modulating polypeptide (GMP), wherein the GMP comprises an actuator moiety linked to a cleavage recognition site, wherein the actuator moiety modulates expression and/or activity of an immune regulatory protein of the lymphocyte, and wherein the immune regulatory protein enhances lymphocyte cytotoxicity and/or reduces a side effect of lymphocyte activation; and

(ii) a chimeric adaptor polypeptide (adaptor) comprising a receptor binding moiety linked to a cleavage moiety, wherein the cleavage moiety is capable of cleaving the cleavage recognition site on the receptor when the receptor binding moiety of the adaptor binds the immune cell signaling domain of the receptor in response to binding of the ligand binding domain of the receptor to a ligand present on the target cell, and wherein the adaptor does not bind a ligand present on the target cell;

wherein the receptor is activatable upon binding to the ligand present on the target cell to recruit the adaptor to the receptor in the lymphocyte, and wherein the recruited adaptor releases the actuator moiety from the GMP of the receptor by action of the cleavage moiety at the cleavage site, thereby inducing death of the target cell.

2. The method of claim 1, wherein modulation of expression and/or activity of the immune regulatory protein enhances cytotoxicity of the lymphocyte compared to a system in which the immune regulatory protein expression and/or activity is not modulated.

3. The method of claim 1, wherein modulation of expression and/or activity of the immune regulatory protein reduces a side effect of lymphocyte activation compared to a system in which the immune regulatory protein expression and/or activity is not modulated, wherein the side effect is hypercytokinemia.

4. The method of claim 1, wherein the actuator moiety up-regulates expression and/or activity of the immune regulatory protein.

5. The method of claim 1, wherein the actuator moiety down-regulates expression and/or activity of the immune regulatory protein.

6. The method of claim 1, wherein the immune regulatory protein is selected from A2AR, B7.1, B7-H3/CD276, B7-H4/B7SI/B7x/Vtcn1, B7-H6, BTLA/CD272, CCR4, CD122, 4-1BB/CD137, CD27, CD28, CD40, CD47, CD70, CISH, CTLA-4/CD152, DR3, GITR, ICOS/CD278, IDO, KIR, LAG-3, OX40/CD134, PD-1/CD279, PD2, PD-L1, PD-L2, TIM-3, and VISTA/Dies1/Gi24/PD-1H (C10orf54).

7. The method of claim 6, wherein the immune regulatory protein is PD-1 or CTLA-4.

8. The method of claim 1, wherein the immune regulatory protein is an immune checkpoint receptor.

9. The method of claim 1, wherein the target cell is a cancer cell.

10. The method of claim 9, wherein the cancer cell is of hematopoietic lineage or a B-cell lymphoma cell.

11. The method of claim 9, wherein the lymphocyte exhibits an enhanced ability to induce death of the target cell as compared to that of a control lymphocyte, wherein the enhanced ability to induce death of the target cell is at least a 1.5-fold increase in induced cell death.

12. The method of claim 9, wherein the cancer cell expresses a ligand indicative of a B-cell lymphoma.

13. The method of claim 1, wherein the ligand is CD19.

14. The method of claim 1, wherein upon binding of the ligand binding domain to the ligand, the receptor undergoes a receptor modification comprising phosphorylation.

15. The method of claim 14, wherein the receptor modification comprises phosphorylation at multiple modification sites, and wherein each modification site is effective to bind a chimeric adaptor polypeptide.

16. The method of claim 1, wherein the enhanced lymphocyte cytotoxicity is evidenced by a reduced size of tumor comprising the target cell.