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KR-102961312-B1 - Allogeneic polypeptide connecting CAR-expressing immune cells and antigen-presenting cells and uses thereof

KR102961312B1KR 102961312 B1KR102961312 B1KR 102961312B1KR-102961312-B1

Abstract

The present invention generally relates to a bispecific polypeptide comprising a first binding domain capable of binding to an antigen-presenting cell (APC) and a second binding domain capable of binding to a T cell, a portion thereof, and uses thereof.

Inventors

  • 커쇼, 마이클
  • 슬라니, 클레어
  • 본 샤이트, 비앙카

Assignees

  • 피터 맥칼룸 캔서 인스티튜트

Dates

Publication Date
20260507
Application Date
20191029
Priority Date
20181030

Claims (20)

  1. As a bispecific polypeptide comprising a first binding domain and a second binding domain, The first binding domain is an antibody or an antigen-binding fragment thereof that specifically binds to an antigen expressed on an endogenous professional antigen-presenting cell (professional APC), and the endogenous professional antigen-presenting cell is not a tumor cell; The antigen expressed by the above-mentioned endogenous specialized antigen-presenting cells is CD40; The second binding domain is an antibody or an antigen-binding fragment thereof that specifically binds to an antigen on an immune cell expressing a chimeric antigen receptor (CAR); and A bispecific polypeptide characterized in that the second binding domain specifically binds to an affinity tag of the CAR.
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  6. In paragraph 1, A bispecific polypeptide characterized in that the above-mentioned specialized antigen-presenting cell is a dendritic cell or a macrophage.
  7. In paragraph 1, A bispecific polypeptide characterized in that the above-mentioned specialized antigen-presenting cell is a dendritic cell.
  8. In paragraph 1, A bispecific polypeptide characterized in that the immune cells are selected from the group consisting of T cells, natural killer cells (NK cells), cytotoxic T lymphocytes, tumor infiltrating lymphocytes (TILs), and regulatory T cells.
  9. In paragraph 8, A bispecific polypeptide characterized in that the immune cell is a T cell.
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  11. In paragraph 1, A bispecific polypeptide characterized in that the affinity tag of the above CAR is selected from the group consisting of c-myc tag, FLAG-tag, HA-tag, His-tag, S-tag, SBP-tag, Strep-tag, eXACT-tag, GST-tag, MBP-tag, and GFP-tag.
  12. In Paragraph 11, A bispecific polypeptide characterized in that the affinity tag of the above CAR is a FLAG-tag.
  13. A nucleic acid encoding the bispecific polypeptide of claim 1.
  14. A vector comprising the nucleic acid of claim 13 operably linked to a regulatory element.
  15. Cells isolated from the human body or non-human cells comprising the vector of claim 14.
  16. A pharmaceutical composition for treating solid tumors, characterized by comprising the bispecific polypeptide of claim 1 and a pharmaceutically acceptable carrier.
  17. As a pharmaceutical composition for the treatment of solid tumors in subjects who have been or are being treated with immune cells expressing a chimeric antigen receptor (CAR), Herein, the pharmaceutical composition comprises a bispecific polypeptide; Herein, the bispecific polypeptide comprises a first binding domain and a second binding domain, wherein the first binding domain specifically binds to an antigen expressed on endogenous specialized antigen-presenting cells of a target, and said endogenous specialized antigen-presenting cells are not tumor cells; Here, the antigen expressed on the above-mentioned endogenous specialized antigen-presenting cells is: (i) not an antigen for a tumor; (ii) not an antigen that is a tumor-associated antigen on the tumor; or (iii) not differentially expressed by the tumor compared to non-tumor cells of the same tissue type; and The above second binding domain specifically binds to the affinity tag of the CAR. A pharmaceutical composition characterized by
  18. As a pharmaceutical composition for the treatment of solid tumors in a subject, Here, the pharmaceutical composition is (i) bispecific polypeptide and (ii) including immune cells expressing a chimeric antigen receptor (CAR); Herein, the bispecific polypeptide comprises a first binding domain and a second binding domain, wherein the first binding domain specifically binds to an antigen expressed on endogenous specialized antigen-presenting cells of a target, and the endogenous specialized antigen-presenting cells are not tumor cells; Here, the antigen expressed on the above-mentioned endogenous specialized antigen-presenting cells is: (i) not an antigen for a tumor; (ii) not an antigen that is a tumor-associated antigen on the tumor; or (iii) not differentially expressed by the tumor compared to non-tumor cells of the same tissue type; and Here, the second binding domain specifically binds to the affinity tag of the CAR. A pharmaceutical composition characterized by
  19. In paragraph 17 or 18, A pharmaceutical composition characterized in that the first binding domain binds to an antigen that is not overexpressed by a tumor compared to non-tumor cells of the same tissue type.
  20. In paragraph 17 or 18, A pharmaceutical composition characterized in that the first binding domain binds to an antigen on an endogenous specialized antigen-presenting cell that is expressed at a higher level by the antigen-presenting cell than by the antigen expressed by the tumor.

Description

Allogeneic polypeptide connecting CAR-expressing immune cells and antigen-presenting cells and uses thereof The present disclosure generally relates to a bispecific polypeptide comprising a first binding domain capable of binding to an antigen-presenting cell (APC) and a second binding domain capable of binding to a T cell, and parts thereof and uses thereof. Adoptive cell transfer (ACT) shows interesting potential for cancer treatment. In ACT, a large number of autologous tumor-reactive T cells are generated in vitro before being re-infused to the patient. Tumor-reactive T cells can be isolated from blood or tumors and can be expanded in vitro through stimulation using peptides and/or cytokines. To date, the most impressive responses to ACT have been found in specific tumor types, such as melanoma, where pretreatment regimens including T cell transfer, IL-2 administration, and chemotherapy and/or whole-body irradiation have demonstrated objective response rates. It was observed in 52 out of 93 patients (56%), 20 out of 93 patients showed a complete response, and 19 out of 20 patients showed a sustained complete response more than 5 years after treatment (Hinrichs et al., 2014, Immunology Reviews, 257(1):56-71). Patients with Epstein-Barr virus (EBV)-associated lymphoproliferative disease after bone marrow transplantation can also benefit from ACT, and virtually all patients achieved complete cure of the disease following adoptive metastasis of EBC-specific T cells (Heslop et al., 2010, Blood, 115(5):925-35). However, the isolation of autologous T cells reactive to other cancer types is rare. Methods to enhance T cell responsiveness to ACT involve genetic modification of patient lymphocytes to generate tumor-responsive T cells for most malignancies, including solid tumors and hematological cancers. Two major approaches to genetic modification involve genes encoding T cell receptors (TCRs) or chimeric antigen receptors (CARs). A CAR consists of an antibody-derived domain fused with a T cell signaling domain that redirects the effector function of T cells toward tumor cells. While both approaches can make T cells tumor-responsive, the non-MHC-restricted CAR approach is more widely applicable to a broader range of patients. Although CARs can take various forms (Kershaw et al., 2013, Nature Reviews Cancer, 13 (8): 525-41), they generally consist of an extracellular domain composed of a single-stranded variable fragment (scFv) of an antibody specific to a tumor-associated antigen (TAA). This scFv is connected to an intracellular region composed of one or more signaling moieties via a hinge and a transmembrane domain. CARs have been developed with specificity for various TAAs, including Her2, CEA, FBP, CD19, and CD209. The most advanced clinical studies have used CD19-specific CARs for the treatment of B-cell leukemia and lymphoma (Kershaw et al., 2013, supra). In 2017, two of these CD19-CAR T-cell therapies received US FDA approval for the treatment of B-cell malignancies (Kymriah, Novartis, and Yescarta, Kite Pharma/Gilead). Despite these results, objective responses reported for solid tumors were less frequent, which may be due to insufficient activation, expansion, and persistence of CAR T cells and/or an immunosuppressive tumor microenvironment. Attempts to optimize this type of therapy have led to the combination of CAR T cells with other therapeutic approaches to overcome tumor-induced immunosuppression, including co-therapy with α-PD-1 monoclonal antibodies, genetic modifications of other signaling and cytokine pathways (Koneru et al., 2015, Oncoimmunology, 4: e994446), the use of adjuvants such as agonistic α-4-1BB monoclonal antibodies (mAB) (Mardiana et al., 2015, Oncoimmunology, 4: e994446), and BiTE (Bispecific T cell participants) (John et al., 2013, Clinical Cancer Research, 19: 5636-46). Some of these approaches have been successfully demonstrated to enable direct interaction between T cells and cancer cells, which can induce T cell expansion in hematopoietic cancer cells, but significant expansion has rarely been observed in the solid tumor environment (Huehls et al., 2015, Immunological Cell Biology, 93 (3): 290-6). Optimization of therapeutic approaches for CAR T cell delivery is often exacerbated by difficulties in the expansion and production of CAR T cells in vitro due to low lymphocyte counts and poor cellular status in severely treated patients (USFDA: KYMRIAH. (tisagenlecleucel), August 30, 2017). Furthermore, even when sufficient donor cells are available, a significant number of cells must be produced to provide an effective dose of these therapies to patients; however, because these are allogeneic cells from healthy donors, graft-versus-host disease (GvHD) can occur due to various issues, such as human leukocyte antigen (HLA) mismatch between the donor and recipient. Therefore, there is an urgent need for the development of new reagents to improve the in vitro and in vivo expansion of