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US-12617860-B2 - Anti-TNFR2 antibody and uses thereof

US12617860B2US 12617860 B2US12617860 B2US 12617860B2US-12617860-B2

Abstract

The invention provides monoclonal antibodies and antigen-binding fragments thereof specific for TNFR2, and methods of using the same to treat cancer or autoimmune disorder, including combination therapy with antagonists of the PD-1/PD-L1 immune checkpoint.

Inventors

  • Shuo Wei
  • Liang Schweizer
  • Francisco Adrian
  • Nicola Arturo Aldo Beltraminelli
  • Pascaline Mary
  • Matthieu Delince
  • Qian Zhang
  • Jennifer Watkins

Assignees

  • HIFIBIO INC.

Dates

Publication Date
20260505
Application Date
20210105

Claims (17)

  1. 1 . An isolated monoclonal antibody, or an antigen-binding fragment thereof, wherein said monoclonal antibody or antigen-binding fragment thereof is specific for human TNFR2, and wherein said monoclonal antibody comprises: (3a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 26, a HCVR CDR2 sequence of SEQ ID NO: 27, and a HCVR CDR3 sequence of SEQ ID NO: 28; and, (3b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 29, a LCVR CDR2 sequence of SEQ ID NO: 30, and a LCVR CDR3 sequence of SEQ ID NO: 31; or (4a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 39, a HCVR CDR2 sequence of SEQ ID NO: 40, and a HCVR CDR3 sequence of SEQ ID NO: 41; and, (4b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 42, a LCVR CDR2 sequence of SEQ ID NO: 43, and a LCVR CDR3 sequence of SEQ ID NO: 44; or (5a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 51, a HCVR CDR2 sequence of SEQ ID NO: 52, and a HCVR CDR3 sequence of SEQ ID NO: 53; and, (5b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 54, a LCVR CDR2 sequence of SEQ ID NO: 55, and a LCVR CDR3 sequence of SEQ ID NO: 56; or (6a) a heavy chain variable region (HCVR), comprising a HCVR CDR1 sequence of SEQ ID NO: 63, a HCVR CDR2 sequence of SEQ ID NO: 64, and a HCVR CDR3 sequence of SEQ ID NO: 65; and, (6b) a light chain variable region (LCVR), comprising a LCVR CDR1 sequence of SEQ ID NO: 66, a LCVR CDR2 sequence of SEQ ID NO: 67, and a LCVR CDR3 sequence of SEQ ID NO: 68, wherein said isolated monoclonal antibody or antigen-binding fragment thereof is a human-mouse chimeric antibody, a humanized antibody, a CDR-grafted antibody, or a resurfaced antibody.
  2. 2 . The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein: (3A) the HCVR sequence is the HCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO: 34; and/or, (3B) the LCVR sequence is the LCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO: 35, or, (4A) the HCVR sequence is the HCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO: 47; and/or, (4B) the LCVR sequence is the LCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO:48, or, (5A) the HCVR sequence is the HCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO: 59; and/or, (5B) the LCVR sequence is the LCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO:60, or, (6A) the HCVR sequence is the HCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO: 71; and/or, (6B) the LCVR sequence is the LCVR sequence of a monoclonal antibody having the heavy chain sequence of SEQ ID NO: 72.
  3. 3 . The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1 , wherein said monoclonal antibody has: (3a) a heavy chain sequence of SEQ ID NO: 34; and/or, (3b) a light chain sequence of SEQ ID NO: 35, or, (4a) a heavy chain sequence of SEQ ID NO: 47; and/or, (4b) a light chain sequence of SEQ ID NO: 48, or, (5a) a heavy chain sequence of SEQ ID NO: 59; and/or, (5b) a light chain sequence of SEQ ID NO: 60, or, (6a) a heavy chain sequence of SEQ ID NO: 71; and/or, (6b) a light chain sequence of SEQ ID NO: 72.
  4. 4 . The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1 , which is a human-mouse chimeric antibody, a humanized antibody, a CDR-grafted antibody, or a resurfaced antibody.
  5. 5 . The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1 , wherein said antigen-binding fragment thereof is an Fab, Fab′, F(ab′)2, single chain Fv or scFv, disulfide linked Fv, IgGΔCH2, minibody, F(ab′)3, tetrabody, triabody, diabody, DVD-Ig, mAb2, (scFv)2, or scFv-Fc.
  6. 6 . The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein said monoclonal antibody or antigen-binding fragment thereof does not substantially cross-react with TNFR1.
  7. 7 . The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein said monoclonal antibody or antigen-binding fragment thereof binds TNFR2 with a K d of less than about 25 nM, 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, or 1 nM.
  8. 8 . The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , which enhances binding between TNFα and TNFR2; enhances TNFα-mediated or -co-stimulated NFκB signaling and/or promotes TCR-activated effector T cell proliferation in the presence of Treg.
  9. 9 . A method of treating cancer in a patient in need thereof, the method comprising administering to the patient an effective amount of the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 .
  10. 10 . The method of claim 9 , which is for treating cancer, wherein the method further comprises administering an antagonist of an immune checkpoint.
  11. 11 . The method of claim 10 , wherein the immune checkpoint is PD-1/PD-L1 immune checkpoint.
  12. 12 . The method of claim 10 , wherein the antagonist of the immune checkpoint is: (a) an antibody or antigen-binding fragment thereof specific for PD-1 or PD-L1, such as cemiplimab, nivolumab, pembrolizumab, avelumab, durvalumab, atezolizumab, KN035, or CK-301; (b) a (non-antibody) peptide inhibitor of PD-1/PD-L1, such as AUNP12; (c) a small molecule inhibitor of PD-L1 such as CA-170; or (d) a macrocyclic peptide such as BMS-986189.
  13. 13 . The method of claim 10 , wherein the cancer is melanoma, breast cancer, colon cancer, cervical cancer, renal cancer, liver cancer (e.g., heptocellular carcinoma), lung cancer (NSCLC), ovarian cancer, skin cancer (e.g., squamous cell carcinoma or basal cell carcinoma), lymphoma, or leukemia.
  14. 14 . The method of claim 10 , further comprising administering to the patient a chemotherapeutic agent, an anti-angiogenesis agent, a growth inhibitory agent, an immune-oncology agent, and/or an anti-neoplastic composition.
  15. 15 . A polynucleotide encoding the heavy chain or the light chain or the antigen-binding portion thereof of claim 1 .
  16. 16 . The polynucleotide of claim 15 , which is codon optimized for expression in a human cell.
  17. 17 . A vector comprising the polynucleotide of claim 15 .

Description

REFERENCE TO RELATED APPLICATIONS This application is a U.S. national stage application filed under 35 U.S.C. § 371 (c), of International Patent Application No. PCT/US2021/012197, filed on Jan. 5, 2021, which claims the benefit of the filing dates of U.S. Provisional Patent Application Nos. 62/957,543, filed on Jan. 6, 2020, and 63/041,234, filed on Jun. 19, 2020, the entire contents of each of the above referenced applications including any drawings and sequence listings are incorporated herein by reference. REFERENCE TO SEQUENCE LISTING The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing file, entitled 131206-00103_SL, was created on Jul. 5, 2022, and is 95,567 bytes in size. The information in electronic format of the Sequence Listing is incorporated herein by reference in its entirety. BACKGROUND OF THE INVENTION Tumor Necrosis Factor Receptor 2 (TNFR2), also known as Tumor Necrosis Factor Receptor Superfamily Member 1B (TNFRSF1B) and CD120b, is a 75-kDa Type I transmembrane protein which contains an extracellular domain (ECD, residues 1-257) with 4 cysteine-rich domains (CRD1 to CRD4), a transmembrane domain (TM, residues 258-287), and an intracellular domain (ICD, residues 288-461) with TRAF2-binding domain. TNFR2 share relatively low sequence identity with the other TNFα receptor—Tumor Necrosis Factor Receptor 1 (TNFR1), with the homology between their extracellular domains being only 28%. TNFR2 binds to the TNFα ligand in a 3:3 trimerization mode. Co-crystal structure of TNFR2 with TNFα has been resolved, and it has been shown that each TNFR2 molecule binds to two TNFα ligands. In addition, TNFα binds TNFR2 with a Kd of 420 pM, about 20 folds weaker than its binding to TNFR1 (Kd=19 nM). Naturally, TNFα preferentially binds to TNFR1 everything else being equal. In normal T cells, TNFα-TNFR2 interaction triggers cell survival signals via the NFkB signaling pathway. In autoimmune T cells, however, TNFα-TNFR2 interaction triggers apoptosis signals via the caspase pathways. Human TNFR2 shows 62% amino acid sequence homology with mouse TNFR2, but it is 97% identical to the rhesus monkey TNFR2. While TNFR1 is ubiquitously expressed, TNFR2 expression is mainly restricted to immune cells, and is predominantly and highly expressed by tumor-infiltrating immunosuppressive CD4+FoxP3+ regulatory T cells (Tregs). Recent studies have shown that TNFR2 plays a crucial role in stimulating the activation and proliferation of Tregs, a major checkpoint of antitumor immune responses (Chen and Oppenheim, Sci Signal 10: eaa12328, 2017). Activation of TNFR2 via its ligand TNFα results in NFkB signaling activation and expansion of TNFR2+ Tregs. TNFR2 is also expressed in CD8 and CD4 Tconv cells, as well as myeloid cells. IN particular, TNFR2 is expressed in exhausted CD8 T cells, similarly to clinically validated immune-checkpoints. T-regulatory cells (Tregs) are a small subset of T-lymphocytes with diverse clinical applications. On the one hand, TNFR2+ Tregs are highly immunosuppressive, with a suppressive activity more potent than that of highly suppressive CD103+ Tregs (J Immunol 179:154-161, 2007; J Immunol 180:6467-6471, 2008). Thus TNFR2+ Tregs can be used in therapy that depends on the immunosuppressive activity of Tregs, such as in transplantation, allergy, asthma, infectious diseases, graft versus host disease (GVHD), and autoimmunity. For example, in experimental GVHD mouse models, CD4+CD25highFoxp3+ thymus-derived Treg depletion could intensify GVHD (Cohen et al., JEM 2002). TNFR2 is also expressed in certain cancers, such as breast cancer, cervical cancer, colon cancer, and renal cancer (Front. Immunol. 9:1170, 2018), and may be involved in immunotolerance in these cancers. The survival and growth of these cancer cells are promoted by ligands of TNFR2 (TNFα). It has been shown that TNFR2 participates in various processes of tumor development by employing different signal pathways in tumor cells. For example, Nuclear Factor-κB (NFκB) is involved in TNFR2-related malignant transformation of epithelial cells. AKT signaling has been shown to be another mediator of TNFR2 in carcinogenesis, tumor growth, and angiogenesis. Meanwhile, Myosin Light-Chain Kinase (MLCK) and Extracellular signal-Regulated Kinase (ERK) are also important for the above-mentioned TNFR2 functions. Thus inhibiting TNFR2 function can inhibit Treg function and increase anti-tumor T cell response in immuno-oncology. Thus, there is a need to develop therapeutic reagents that allow one to either enhance the immunosuppressive function of Tregs to treat autoimmune disorders through stimulating TNFR2 function on TNFR2+ Tregs, or to inhibit TNFR2 activation for treating diseases such as cancer. SUMMARY OF THE INVENTION In one aspect, the invention provides an isolated monoclonal antibody, or an antigen-binding fragment thereof, wherein said monoclonal antibody or antigen-binding fragment thereof i