CD122 binding agents and methods of use thereof

Abstract

The present invention provides a CD122 binding agent comprising an immunoglobulin heavy chain polypeptide and an immunoglobulin light chain polypeptide, as well as related compositions and methods of making and using the same.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 423,416, filed on November 7, 2022, the entire disclosure of which is hereby incorporated by reference.

[0003] Incorporation by Reference of Electronically Submitted Materials

[0004] The computer-readable nucleotide / amino acid sequence listing submitted concurrently and identified as follows is incorporated by reference in its entirety: a 214,653-byte ASCII (text) file named "769570_ST.26.xml", created on November 2, 2023. Background Art

[0005] Interleukin-2 (IL-2) and interleukin-15 (IL-15) are two important cytokines involved in immune system regulation. IL-2 participates in regulatory T cell (Treg) cell development and homeostasis, while IL-15 regulates NK and NK_T cell development and memory T cell survival. The receptors of IL-2 (IL-2R) and IL-15 (IL-15R) each include three subunits named as α, β and γ. Although IL-2R and IL-15R have unique α subunits (respectively, CD25 and CD125), these two receptors have common β subunits and γ subunits (CD122 and CD132).

[0006] IL-2 and IL-15 signaling are involved in pathogenic immune system responses. Therefore, there is a need for CD122 binding agents that can modulate this signaling. Summary of the Invention

[0007] Provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 113 or at least the CDR region thereof, and the immunoglobulin light chain variable region comprises SEQ ID NO: 114 or at least the CDR region thereof.

[0008] In another aspect, the present disclosure provides a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 69 or at least the CDR region thereof, and the immunoglobulin light chain variable region comprises SEQ ID NO: 77 or at least the CDR region thereof.

[0009] In another aspect, the present disclosure provides a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 150 or at least the CDR region thereof, and the immunoglobulin light chain variable region comprises SEQ ID NO: 151 or at least the CDR region thereof.

[0010] In another aspect, the present disclosure provides a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 24 or at least the CDR region thereof, and the immunoglobulin light chain variable region comprises SEQ ID NO: 35 or at least the CDR region thereof.

[0011] The present disclosure further provides a method for suppressing the immune response in a mammal and treating an inflammatory disorder or an autoimmune disorder in the mammal by administering the CD122 binding agent to the mammal. Also provided are nucleic acid sequences encoding the aforementioned immunoglobulin polypeptides and binding agents, as well as related compositions and methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 Shown are the results of a cell-based receptor binding assay demonstrating the ability of anti-CD122 antibodies to bind to CHO-K1 cells expressing human IL-2βγ.

[0013] Figure 2 Shown are the results of a cell-based receptor binding assay demonstrating the ability of anti-CD122 antibodies to bind to CHO-K1 cells expressing cynomolgus IL-2βγ.

[0014] Figure 3 Shown are the results of a cell-based functional assay demonstrating the ability of anti-CD122 antibodies to inhibit IL-2-induced pSTAT5 luciferase activity in HEK cells.

[0015] Figure 4 Shown are the results of a CTG (cell titer glo)-based proliferation assay demonstrating the ability of humanized anti-CD122 IgG4 antibodies to inhibit IL-2 or IL-15-induced proliferation of PHA-stimulated primary human cell blasts.

[0016] Figure 5 The binding activity of anti-CD122 IgG4 antibody to CHO-K1 cells expressing human or cynomolgus monkey IL-2βγ is shown.

[0017] Figure 6Shown are the effects of anti-CD122 IgG4 antibodies on IL-2- or IL-15-induced proliferation of purified primary human NK cells.

[0018] Figure 7 Shown are the effects of humanized anti-CD122 IgG4 antibodies on IL-2- or IL-15-induced proliferation and survival of primary human NK cells, gated within PBMC cultures.

[0019] Figure 8 Shown are the effects of humanized anti-CD22 IgG4 antibodies on the binding of IL-2 or IL-15 to HEK cells expressing human IL-2βγ.

[0020] Figure 9 Shown are the effects of humanized anti-CD122 IgG4 antibodies on IL-15 or IL-15 / IL1-5RA-induced proliferation of primary human T cell blasts.

[0021] Figure 10 Shown are the effects of humanized anti-CD122 IgG4 antibodies on IL-15 or IL-15 / IL1-5RA-induced proliferation of primary human NK cells.

[0022] Figure 11 Shown are the effects of humanized anti-CD122 IgG1 LALA antibody on IL-2- or IL-15-induced proliferation of primary human NK cells.

[0023] Figure 12 Shown are the effects of humanized anti-CD122 IgG1 LALA antibody on IL-2- or IL-15-induced proliferation of primary human pan T cells.

[0024] Figure 13 Shown are the effects of humanized anti-CD122 antibody alone or in combination with CTLA4-Ig in a xenogeneic NSG (huIL-15) / Hu-PBMC GvHD model.

[0025] Figure 14 Shown are the effects of humanized anti-CD122 antibodies in a xenogeneic NSG (huIL-15Tg) / Hu-PBMC graft-versus-host disease (GvHD) model. DETAILED DESCRIPTION

[0026] The present invention provides a kind of CD122 binding agent.The CD122 binding agent comprises immunoglobulin heavy chain variable region and immunoglobulin light chain variable region, and the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region each comprise three complementary determining regions (CDR), and the three complementary determining regions are commonly referred to as CDR1, CDR2 or CDR3.CDR area can also refer to using "H" or "L" in the term to refer to, to represent heavy chain or light chain, i.e., CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 or CDRL3, respectively. The CDRs of a given Ig sequence can be identified by any of several conventional numbering schemes, such as Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH (1991); Chothia et al., "Canonical Structures for the Hypervariable Regions of Immunoglobulins," J. Mol. Biol., 196:901-917 (1987); Al-Lazikani et al., "Standard Conformations for the Canonical Structures of Immunoglobulins," J. Mol. Biol., 196:901-917 (1987); Al-Lazikani et al., "Standard Conformations for the Canonical Structures of Immunoglobulins," Abhinandan et al., “Analysis and Improvements to Kabat and Structurally Correct Numbering of Antibody Variable Domains,” Mol. Immunol.), 45:3832–3839 (2008); Lefranc et al., “The IMGT unique numbering for immunoglobulins, T cell receptors and Ig-like domains,” The Immunologist, 7:132–136 (1999); Lefranc et al., “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and I superfamily V-like domains,” Dev. Comp. Immunol., 27:55–77 (2003); and Honegger et al., “Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool.” tool),"Journal of Molecular Biology 309:657–670 (2001).

[0027] Provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 113 or at least the CDR regions thereof, and the immunoglobulin light chain variable region comprises SEQ ID NO: 114 or at least the CDR regions thereof, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo.

[0028] Also provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein the immunoglobulin heavy chain variable region comprises one of SEQ ID NOs: 37-69, or at least the CDRs thereof; and the immunoglobulin light chain variable region comprises the CDRs of any one of SEQ ID NOs: 70-77, wherein the CDRs are as determined according to any of the various known immunoglobulin numbering schemes (e.g., Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo).

[0029] Also provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: 37-69; and the immunoglobulin light chain variable region comprises an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: 37-69; Any of NOs:70-77 has an amino acid sequence with at least 80%, 85% or 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity). In some embodiments, the immunoglobulin heavy chain variable region comprises the CDRs of any one of SEQ ID NOs: 37-69 and has an amino acid sequence that has at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: 37-69; and the immunoglobulin light chain variable region comprises the CDRs of any one of SEQ ID NOs: 70-77 and has an amino acid sequence that has at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: Any of NOs:70-77 has an amino acid sequence with at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity); wherein the CDRs are as defined according to Kabat, Chothia, Martin (enhanced Chothia), IGMT or Aho.

[0030] Also provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 69; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 69; and / or at least the CDRs of SEQ ID NO: 69, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or Aho; and the immunoglobulin light chain variable region comprises SEQ ID NO: 77; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 69; NO:77 has an amino acid sequence with at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity); and / or at least the CDR regions of SEQ ID NO:77, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo.

[0031] In some embodiments, the CD122 binding agent comprises a heavy chain variable region of SEQ ID NO: 69 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by Kabat. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 69 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by Chothia. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 69 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by Martin. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 69 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by IGMT. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 69 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by AHo. By further example, a CD122-binding agent can comprise an immunoglobulin heavy chain comprising SEQ ID NO: 115 and an immunoglobulin light chain comprising SEQ ID NO: 116, or amino acid sequences having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NOs: 115 and 116, respectively, optionally wherein the sequences retain the identity of SEQ ID NOs: The heavy chain CDRs and light chain CDRs of NOs: 115 and 116, wherein the CDRs are as determined according to any of the various known immunoglobulin numbering schemes (e.g., Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo).

[0032] Also provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 68; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 68; and / or at least the CDRs of SEQ ID NO: 68, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or Aho; and the immunoglobulin light chain variable region comprises SEQ ID NO: 77; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: NO:77 has an amino acid sequence with at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity); and / or at least the CDR regions of SEQ ID NO:77, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo.

[0033] In some embodiments, the CD122 binding agent comprises a heavy chain variable region of SEQ ID NO: 68 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by Kabat. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 68 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by Chothia. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 68 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by Martin. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 68 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by IGMT. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 68 and a light chain variable region of SEQ ID NO: 77, or at least its CDRs, as determined by AHo. By further example, a CD122-binding agent can comprise an immunoglobulin heavy chain comprising SEQ ID NO: 165 and an immunoglobulin light chain comprising SEQ ID NO: 116, or amino acid sequences having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NOs: 165 and 116, respectively, optionally wherein the sequences retain the identity of SEQ ID NOs: The heavy chain CDRs and light chain CDRs of NOs: 165 and 116, wherein the CDRs are as determined according to any of the various known immunoglobulin numbering schemes (e.g., Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo).

[0034] In yet another aspect, the present disclosure provides a CD122-binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region of the CD122-binding agent comprise the following CDRs, when determined according to Kabat numbering: CDRH1 comprising SEQ ID NO: 108; CDRH2 comprising SEQ ID NO: 109; CDRH3 comprising SEQ ID NO: 98; CDRL1 comprising SEQ ID NO: 110; CDRL2 comprising SEQ ID NO: 111; and CDRL3 comprising SEQ ID NO: 112.

[0035] In some embodiments, when determined according to Kabat numbering, the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region comprise the following CDRs: CDRH1 comprises any one of SEQ ID NOs: 78-93; CDRH2 comprises any one of SEQ ID NOs: 94-97; CDRH3 comprises SEQ ID NO: 98; CDRL1 comprises any one of SEQ ID NOs: 99-102; CDRL2 comprises SEQ ID NOs: 103 or 104; and CDRL3 comprises any one of SEQ ID NOs: 105-107.

[0036] In another aspect, the present disclosure provides a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 150, or at least the CDR regions thereof, and the immunoglobulin light chain variable region comprises SEQ ID NO: 151, or at least the CDR regions thereof, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo.

[0037] Also provided is a CD122-binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein the immunoglobulin heavy chain variable region comprises any one of SEQ ID NOs: 1-24, or at least the CDRs thereof, and the immunoglobulin light chain variable region comprises any one of SEQ ID NOs: 25-36, or at least the CDRs thereof, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or AHo.

[0038] Also provided herein is a CD122-binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: 1-24; and the immunoglobulin light chain variable region comprises an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: 1-24; NOs:25-36 at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity). In some embodiments, the immunoglobulin heavy chain variable region comprises the CDRs of any one of SEQ ID NOs: 1-24 and has an amino acid sequence that has at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: 1-24; and the immunoglobulin light chain variable region comprises the CDRs of any one of SEQ ID NOs: 25-36 and has an amino acid sequence that has at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to any one of SEQ ID NOs: any one of NOs:25-36 having an amino acid sequence with at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity); wherein the CDRs are as defined according to Kabat, Chothia, Martin (enhanced Chothia), IGMT, or Aho.

[0039] Also provided is a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 24; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 24; and / or at least the CDR regions of SEQ ID NO: 24, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or Aho; and the immunoglobulin light chain variable region comprises SEQ ID NO: 35; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 24; NO:35 has an amino acid sequence having at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity); and / or at least the CDR regions of SEQ ID NO:35, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo.

[0040] In some embodiments, the CD122 binding agent comprises a heavy chain variable region of SEQ ID NO: 24 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by Kabat. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 24 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by Chothia. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 24 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by Martin. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 24 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by IGMT. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 24 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by AHo. By way of further example, the CD122 binding agent may comprise: an immunoglobulin heavy chain comprising SEQ ID NO: 152; an immunoglobulin light chain comprising SEQ ID NO: 153; or NOs: 152 and 153 have amino acid sequences that have at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity), optionally wherein the sequences retain the heavy and light chain CDRs of SEQ ID NOs: 152 and 153 as determined according to any of the various known immunoglobulin numbering schemes (e.g., Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo).

[0041] Also provided is a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 23; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 23; and / or at least the CDR regions of SEQ ID NO: 23, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or Aho; and the immunoglobulin light chain variable region comprises SEQ ID NO: 35; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 23; NO:35 has an amino acid sequence having at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity); and / or at least the CDR regions of SEQ ID NO:35, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo.

[0042] In some embodiments, the CD122 binding agent comprises a heavy chain variable region of SEQ ID NO: 23 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by Kabat. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 23 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by Chothia. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 23 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by Martin. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 23 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by IGMT. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 23 and a light chain variable region of SEQ ID NO: 35, or at least its CDRs, as determined by AHo. By way of further example, the CD122 binding agent may comprise: an immunoglobulin heavy chain comprising SEQ ID NO: 157; an immunoglobulin light chain comprising SEQ ID NO: 153; or NOs: 157 and 153 have amino acid sequences that have at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity), optionally wherein the sequences retain the heavy and light chain CDRs of SEQ ID NOs: 157 and 153 as determined according to any of the various known immunoglobulin numbering schemes (e.g., Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo).

[0043] Also provided is a CD122-binding agent comprising an immunoglobulin heavy chain and an immunoglobulin light chain, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 16; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 16; and / or at least the CDR regions of SEQ ID NO: 16, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT, or Aho; and the immunoglobulin light chain variable region comprises SEQ ID NO: 28; an amino acid sequence having at least 80%, 85%, or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to SEQ ID NO: 16; NO:28 has an amino acid sequence having at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity); and / or at least the CDR regions of SEQ ID NO:28, wherein the CDR regions are as defined by Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo.

[0044] In some embodiments, the CD122 binding agent comprises a heavy chain variable region of SEQ ID NO: 16 and a light chain variable region of SEQ ID NO: 28, or at least its CDRs, as determined by Kabat. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 16 and a light chain variable region of SEQ ID NO: 28, or at least its CDRs, as determined by Chothia. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 16 and a light chain variable region of SEQ ID NO: 28, or at least its CDRs, as determined by Martin. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 16 and a light chain variable region of SEQ ID NO: 28, or at least its CDRs, as determined by IGMT. In some embodiments, the antibody comprises a heavy chain variable region of SEQ ID NO: 16 and a light chain variable region of SEQ ID NO: 28, or at least its CDRs, as determined by AHo. By way of further example, the CD122 binding agent may comprise: an immunoglobulin heavy chain comprising SEQ ID NO: 159; an immunoglobulin light chain comprising SEQ ID NO: 161; or NOs: 159 and 161 have amino acid sequences that have at least 80%, 85% or 90% sequence identity (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity), optionally wherein the sequences retain the heavy and light chain CDRs of SEQ ID NOs: 159 and 161 as determined according to any of the various known immunoglobulin numbering schemes (e.g., Kabat, Chothia, Martin (enhanced Chothia), IGMT or AHo).

[0045] According to another aspect, the present disclosure provides a CD122-binding agent comprising a heavy chain immunoglobulin variable region and a light chain immunoglobulin variable region, wherein the heavy chain immunoglobulin variable region and the light chain immunoglobulin variable region have the following CDRs, when determined by Kabat numbering: CDRH1, wherein the CDRH1 comprises SEQ ID NO: 146; CDRH2, wherein the CDRH2 comprises SEQ ID NO: 147; and CDRH3, wherein the CDRH3 comprises SEQ ID NO: 148; CDRL1, wherein the CDRL1 comprises SEQ ID NO: 138; CDRL2, wherein the CDRL2 comprises SEQ ID NO: 139; and CDRL3, wherein the CDRL3 comprises SEQ ID NO: 149. In some embodiments, CDRH1 comprises any one of SEQ ID NOs: 117-126; CDRH2 comprises any one of SEQ ID NOs: 127-135; CDRH3 comprises SEQ ID NOs: 136 or 137; CDRL1 comprises SEQ ID NO: 138; CDRL2 comprises SEQ ID NO: 139; and CDRL3 comprises any one of SEQ ID NOs: 140-145.

[0046] Further provided is a CD122-binding agent comprising a heavy chain variable region and a light chain variable region shown in Table 1A or 1B, or at least CDRs thereof, as determined by Kabat, Chothia, Martin (enhanced Chothia), IGMT or Aho. Also provided is a CD122-binding agent comprising the CDR regions shown in Tables 1C and 1D when determined according to Kabat numbering.

[0047] As described herein, sequence "identity" can be determined by comparing the nucleic acid or amino acid sequence of interest to a reference nucleic acid or amino acid sequence. Percent identity is the number of nucleotides or amino acid residues that are identical (same / identical) between the sequence of interest and the reference sequence divided by the length of the longest sequence (that is, the length of the longer side in the sequence of interest or the reference sequence). A variety of mathematical algorithms for obtaining optimal comparisons and calculating the identity between two or more sequences are known and are incorporated into multiple available software programs. Examples of such programs include CLUSTAL-W, T-Coffee, and ALIGN (for nucleic acid and amino acid sequence alignments), BLAST programs (e.g., BLAST 2.1, BL2SEQ, and subsequent versions thereof), and FASTA programs (e.g., FASTA3x, FASTM, and SSEARCH) (for sequence alignment and sequence similarity searches). Sequence alignment algorithms are also disclosed in, for example, Altschul et al., J. Mol. Biol., 215(3):403-410 (1990); Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10):3770-3775 (2009); Durbin et al., eds., Biological Sequence Analysis: Probabalistic Models of Proteins and Nucleic Acids, Cambridge University Press, Cambridge, UK (2009); Soding, Bioinformatics, 21(7):951-960 (2005); Altschul et al., Nucleic Acids Res., 1996. Res., 25(17):3389-3402 (1997); and Gusfield, Algorithms on Strings, Trees and Sequences, Cambridge University Press, Cambridge UK (1997).

[0048] The variation of sequence identity can be realized by adding, replacing or lacking one or more amino acid residues.Amino acid " replacement " or " replacement " refers to that an amino acid at a given position or residue in a polypeptide sequence is replaced by another amino acid at the same position or residue.Amino acid replacement or replacement can be conservative, semiconservative or nonconservative, depending on whether the replacement is carried out by an amino acid residue with similar properties to the replaced residue. The functional mode for defining the common characteristics between independent amino acids is the normalized frequency (Schulz and Schirmer, " Principles of Protein Structure ", Springer-Verlag, New York (1979)) of the amino acid changes between the corresponding proteins of the analysis homologous organisms. According to this type of analysis, amino acid groups can be defined, wherein the amino acid in the group preferentially replaces each other, and thus most similar to each other (Schulz and Schirmer, the same) in terms of its impact on overall protein structure.

[0049] Amino acids can be broadly grouped as "aromatic" or "aliphatic." Aromatic amino acids include an aromatic ring. Examples of "aromatic" amino acids include histidine (H or His), phenylalanine (F or Phe), tyrosine (Y or Tyr), and tryptophan (W or Trp). Non-aromatic amino acids are broadly grouped as "aliphatic." Examples of "aliphatic" amino acids include glycine (G or Gly), alanine (A or Ala), valine (V or Val), leucine (L or Leu), isoleucine (I or Ile), methionine (M or Met), serine (S or Ser), threonine (T or Thr), cysteine ​​(C or Cys), proline (P or Pro), glutamic acid (E or Glu), aspartic acid (A or Asp), asparagine (N or Asn), glutamine (Q or Gln), lysine (K or Lys), and arginine (R or Arg).

[0050] Aliphatic amino acids can be subdivided into four subgroups. The "large aliphatic nonpolar subgroup" consists of valine, leucine, and isoleucine. The "aliphatic weakly polar subgroup" consists of methionine, serine, threonine, and cysteine. The "aliphatic polar / charged subgroup" consists of glutamic acid, aspartic acid, asparagine, glutamine, lysine, and arginine. The "small residue subgroup" consists of glycine and alanine. The group of charged / polar amino acids can be subdivided into three subgroups: the "positively charged subgroup" consisting of lysine and arginine, the "negatively charged subgroup" consisting of glutamic acid and aspartic acid, and the "polar subgroup" consisting of asparagine and glutamine.

[0051] Aromatic amino acids can be subdivided into two subgroups: the "nitrogen ring subgroup" consisting of histidine and tryptophan, and the "phenyl subgroup" consisting of phenylalanine and tyrosine.

[0052] Examples of conservative amino acid substitutions include substitutions of amino acids within the subgroups described above, such as substitutions of lysine for arginine, or vice versa, to maintain a positive charge; substitutions of glutamic acid for aspartic acid, or vice versa, to maintain a negative charge; substitutions of serine for threonine to maintain a free -OH; and substitutions of glutamine for asparagine to maintain a free -NH. "Semi-conservative mutations" include substitutions of amino acids within the same group listed herein but not within the same subgroup. For example, substitutions of aspartic acid for asparagine or asparagine for lysine involve amino acids within the same group but within different subgroups. "Non-conservative mutations" involve substitutions of amino acids between different groups, such as substitutions of lysine for tryptophan, or substitutions of phenylalanine for serine, etc.

[0053] In some embodiments, the CD122 binding agent can include the immunoglobulin heavy chain variable region and immunoglobulin light chain variable region or complete heavy chain and light chain polypeptides provided herein, consisting essentially of or consisting of. The CD122 binding agent can be any type of molecule or construct comprising at least a specified immunoglobulin heavy chain variable region and immunoglobulin light chain variable region. Therefore, the CD122 binding agent can be, for example, a complete immunoglobulin or antibody, as described herein, or an antigen-binding (CD122 binding) immunoglobulin or antibody "fragment". The term "fragment" used with respect to antibodies or immunoglobulins means any molecule or construct comprising some portion of an immunoglobulin or antibody and bound to a target antigen. This fragment will typically comprise at least a portion of the heavy chain variable region and light chain variable region including CDR, and may also include a portion of a constant region, optionally together with other elements (e.g., linkers, etc.) that are not typically part of an immunoglobulin or antibody. Examples of such "fragments" include, but are not limited to: (i) a Fab fragment, which is a fragment consisting of a V L 、V H 、C L and CH1 domains; (ii) F(ab')2, which is a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; (iii) Fv fragment, which consists of the V domains of a single arm of an antibody. L domain and V H domain composition; (iv) Fab' fragments, which are produced by cleaving the disulfide bonds of the F(ab')2 fragment using mild reducing conditions; (v) diabodies; (vi) single-chain variable regions (scFv); and (vii) disulfide-stabilized Fv fragments (dsFv).

[0054] In some embodiments, the CD122 binding agent comprises an immunoglobulin heavy chain constant region, such as a fragment crystallizable (F c ) region or part thereof. The Fc region can belong to any Ig class / subclass (IgA (IgA1, IgA2), IgD, IgE, IgG (IgG1, IgG2, IgG3 and IgG4), IgM, including variants thereof). In a specific embodiment, the CD122 binding agent comprises an Fc region that binds to an Fc receptor of an antigen presenting cell (e.g., a dendritic cell, a macrophage, a Langerhans cell, or a B cell). The Fc receptor can be an Fcγ receptor (FcγR), such as FcγRI (CD64), FcγRIIA (CD32), FcγRIIB (CD32), FcγRIIIA (CD16a), FcγRIIIB (CD16b). In one embodiment, the CD122 binding agent comprises an Fc region that binds to FcγR, such as IgG1. Thus, in some embodiments, the CD122 binding agent is a "complete" or "complete" Ig (i.e., an antibody). In another embodiment, the CD122 binding agent is an IgG antibody, particularly an IgG1 antibody. In another embodiment, the CD122 agent is an IgG antibody, particularly an IgG4 antibody.

[0055] In some embodiments, CD122 binding agents include reduced (including substantially or completely eliminated) binding to one or more (or all) Fc γ receptors and thus reduced immune effector function Fc region.CD122 binding agents can include, for example, IgG Fc regions (e.g., IgG1 or IgG4) with the following: a mutation that reduces effector function (e.g., removing the glycosylation sites of the Fc N connections in human IgG1), a leucine to glutamic acid substitution at position 235 of IgG1 Fc or other modifications of hinge region positions 234-237; double mutations Leu234Ala and Leu235Ala ("LALA" mutations); substitution P329G in IgG1; or both P329G and LALA mutations. In some embodiments, CD122 binding agents include only IgG1 Fc with LALA mutations or further IgG1 Fc with P329G.

[0056] The CD122 binding agent of separation can also be an antibody conjugate.In this respect, the CD122 binding agent of the separation can be a conjugate comprising a CD122 binding agent (e.g., an anti-CD122 antibody or antibody fragment) and another biologically active portion. For example, the CD122 binding agent can be coupled with a peptide, a fluorescent molecule or a chemotherapy agent, particularly with an agent that can be used to suppress an immune response.

[0057] The CD122 binding agent can be a human antibody, a non-human antibody or a chimeric antibody or can be obtained therefrom."Chimeric" means an antibody or fragment thereof comprising both a human region and a non-human region. Preferably, the CD122 binding agent of separation is a humanized antibody. "Humanized" antibody is a monoclonal antibody comprising a human antibody scaffold and at least one CDR obtained from or derived from a non-human antibody. Non-human antibodies include antibodies isolated from any non-human animal, such as rodents (e.g., mice or rats). Humanized antibodies can include one, two or three CDRs obtained from or derived from non-human antibodies. In a preferred embodiment of the present invention, the CDRH3 of the CD122 binding agent is obtained from or derived from a mouse monoclonal antibody, and the remaining variable regions and constant regions of the CD122 binding agent of the present invention are obtained from or derived from human monoclonal antibodies.

[0058] Human, non-human, chimeric, or humanized antibodies can be obtained by any means, including in vitro sources (e.g., hybridomas or cell lines that recombinantly produce antibodies) and in vivo sources (e.g., rodents). Methods for producing antibodies are known in the art and are described, for example, in and Milstein, Eur. J. Immunol., 5:511-519 (1976); Harlow and Lane (eds.), Antibodies: A Laboratory Manual, CSH Press (1988), and Janeway et al. (eds.), Immunobiology, 5th ed., Garland Publishing, New York, NY (2001); Starkie et al., PLoS One, 11(3):e0152282 (2016). In certain embodiments, human antibodies or chimeric antibodies can be produced using transgenic animals (e.g., mice) in which one or more endogenous immunoglobulin genes are replaced by one or more human immunoglobulin genes. Examples of transgenic mice in which endogenous antibody genes are effectively replaced by human antibody genes include, but are not limited to, the Medarex HUMAB-MOUSE TM 、Kirin TC MOUSE TM and Kyowa Kirin KM-MOUSE TM(See, eg, Lonberg, Nat. Biotechnol., 23(9): 1117-25 (2005) and Lonberg, Handb. Exp. Pharmacol., 181: 69-97 (2008)). Humanized antibodies can be produced using any suitable method known in the art (see, e.g., An, Z. (ed.), Therapeutic Monoclonal Antibodies: From Bench to Clinic, John Wiley & Sons, Inc., Hoboken, New Jersey (2009)), including, for example, grafting non-human CDRs onto human antibody scaffolds (see, e.g., Kashmiri et al., Methods, 36(1):25-34 (2005); and Hou et al., J. Biochem., 144(1):115-120 (2008)). In one embodiment, humanized antibodies can be produced using, for example, the methods described in U.S. Patent Application Publication No. 2011 / 0287485A1.

[0059] The CD122 binding agent can have any suitable affinity for human CD122. The term "affinity" refers to the equilibrium constant for the reversible binding of two agents and is expressed as the dissociation constant (K D ). The affinity of the binding agent for the ligand (such as the affinity of the antibody for the epitope) can be, for example, from about 1 picomolar (pM) to about 100 micromolar (μM) (e.g., from about 1 picomolar (pM) to about 1 nanomolar (nM), from about 1 nM to about 1 micromolar (μM), or from about 1 μM to about 100 μM). In one embodiment, the CD122 binding agent can have a K of less than or equal to 1.5 nM (e.g., 1.4 nM, 1.3 nM, 1.2 nM, 1.0 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM, 0.05 nM, 0.025 nM, 0.01 nM, 0.001 nM, or a range defined by any two of the foregoing values). DIn another embodiment, the CD122 binding agent can be with a K of less than or equal to 200pM (e.g., 190pM, 175pM, 150pM, 125pM, 110pM, 100pM, 90pM, 80pM, 75pM, 60pM, 50pM, 40pM, 30pM, 25pM, 20pM, 15pM, 10pM, 5pM, 1pM or a range defined by any two values ​​of the aforementioned values). D Binds to CD122. In some embodiments, the affinity of the cross-reactivity of the CD122 binding agent to cynomolgus monkey CD122 is within any of the aforementioned ranges discussed for human CD122. The affinity of the immunoglobulin for the antigen or epitope of interest can be measured using any art-recognized assay. Such methods include, for example, fluorescence activated cell sorting (FACS), separable beads (e.g., magnetic beads), surface plasmon resonance (SPR), solution phase competition Antigen panning and / or ELISA (see, eg, Janeway et al. (eds.), Immunobiology, 5th ed., Garland Publishers, New York, NY (2001)).

[0060] IL-2Rβ (CD122) is a common β receptor subunit shared by IL-15 and IL-2. IL-15 and IL-2 signaling mediate the survival and maintenance of tissue-resident memory T cells (TRMs) and NK cell subsets. The presence of long-lived and persistent TRMs has been shown to be a pathogenic driver of tissue-specific immune-mediated inflammation in the skin of dermatological diseases, where well-defined inflammation frequently recurs. TRMs have also been observed in other tissue-specific inflammatory conditions, including gastrointestinal and rheumatic diseases.

[0061] In some embodiments, CD122 binding agents are combined with CD122 and at least partially (or completely) suppress IL-15 signal transduction.Alternatively or in addition, CD122 binding agents are combined with CD122 and at least partially (or completely) suppress IL-2 signal transduction.For example, in some embodiments, CD122 binding agents at least partially (or completely) suppress IL-15 signal transduction, and suppress IL-2 signal transduction by low-affinity IL-2 receptors (including CD122 and common γ subunit CD132), without eliminating (or not suppressing) IL-2 signal transduction by high-affinity IL-2 receptors (including CD122, CD132 and IL-2 α receptor subunit CD25).CD122 binding agents can be in conjunction with CD122 on any cell type expressing CD122 (such as regulatory T cells (Treg), memory T cells, CD56+ T cells (NKT cells), innate lymphocytes (ILC) and γδT cells). Without wishing to be bound by any particular theory or mechanism of action, it is believed that inhibiting IL-15 signaling (in whole or in part) and inhibiting IL-2 signaling (in whole or in part) through the low-affinity IL-2 receptor expressed on NK cells and T cells, without eliminating IL-2 signaling through the high-affinity IL-2 receptor expressed on regulatory T cells, can provide additional anti-inflammatory benefits by reducing the number of NK cells and pathogenic T cells while preserving or potentially increasing the number of regulatory T cells.

[0062] Use / treatment methods

[0063] CD122 binding agents are considered to be particularly useful for suppressing IL-15 signal transduction and suppressing IL-2 signal transduction, particularly by low-affinity IL-2 receptors (comprising CD122 and common gamma subunit CD132). Thus, the binding agent can be used for treating a disease, disorder or its symptom caused by or promoted by IL-15 signal transduction or IL-2 signal transduction, particularly by a method for treating a disease or disorder associated with low-affinity IL-2 receptors. For example, CD122 binding agents can be used for suppressing inflammatory response or immune response or treating a disease or disorder associated therewith, particularly characterized by a disease or disorder of pathogenic memory T cells (including tissue-resident memory T cells (TRM cells)) and NK cells. In some embodiments, the signal transduction to regulatory T cells (Treg) is retained in whole or in part (that is, compared to such signal transduction in the absence of CD122 binding agents, in the presence of CD122 binding agents, Treg signal transduction is not completely blocked, or is not significantly suppressed, or is not suppressed at all). Negative regulation of the immune system is used synonymously with immunosuppression here. It will be appreciated that in some cases, a CD122 binding agent can be administered prior to the onset of symptoms (e.g., prior to exposure to an antigen that triggers an immune response) in order to prevent, suppress, or reduce the severity of the immune response following introduction of the antigen.

[0064] In some aspects, CD122 binding agents can be used to treat inflammatory disorders or autoimmune disorders, or otherwise inhibit or suppress an immune response or inflammatory response. Examples of inflammatory or autoimmune disorders include, for example, infections (viral, bacterial, fungal, and parasitic), endotoxin shock associated with infection, arthritis, rheumatoid arthritis including TNF-refractory rheumatoid arthritis, Sjogren's Syndrome, asthma, chronic obstructive pulmonary disease (COPD), pelvic inflammatory disease, Behcet's disease, Alzheimer's disease, inflammatory bowel disease including Crohn's disease and ulcerative colitis, Peyronie's disease, celiac disease, gallbladder disease, pilonidal disease, peritonitis, psoriasis, psoriatic arthritis, contact hypersensitivity, allergic diseases, eosinophilia, eosinophilic esophagitis, vasculitis, antineutrophil cytoplasmic antibody-associated (ANCA) vasculitis, surgical adhesions, stroke, type 1 diabetes, Lyme disease, disease), arthritis, meningoencephalitis, autoimmune or noninfectious uveitis, immune-mediated inflammatory disorders of the central and peripheral nervous systems (such as multiple sclerosis), lupus (such as systemic lupus erythematosus and chronic discoid lupus erythematosus) and Guillain-Barre syndrome, atopic dermatitis, polymyositis, dermatomyositis, autoimmune hepatitis, fibrosing alveolitis, Graves' disease, IgA nephropathy, idiopathic thrombocytopenic purpura, Meniere's disease disease), pemphigus, pemphigoid, primary biliary cholangitis, hepatitis, sarcoidosis, scleroderma (localized, systemic, and progressive systemic), granulomatosis with polyangiitis, other autoimmune disorders, cholangitis, pancreatitis, trauma (surgical), graft-versus-host disease, transplant rejection, heart disease including ischemic disease such as myocardial infarction and atherosclerosis, periarteritis nodosa (polyarteritis nodosa and microscopic polyangiitis), allergic granulomatous vasculitis, hypersensitivity vasculitis, aortitis syndrome (Takayasu arteritis), temporal arteritis, intravascular coagulation, bone resorption, osteoporosis, osteoarthritis, periodontitis including chronic periodontitis, hypochlorhydria, Still's disease, Cogan's syndromesyndrome), RS3PE, polymyalgia rheumatica, fibromyalgia syndrome, antiphospholipid antibody syndrome, eosinophilic fasciitis, Guillain-Barré syndrome, myasthenia gravis, chronic atrophic gastritis, Goodpasture's syndrome, rapidly progressive glomerulonephritis, megaloblastic anemia, hemolytic anemia, autoimmune neutropenia, Hashimoto's thyroiditis, autoimmune adrenal insufficiency, primary hypothyroidism, idiopathic Addison's disease (chronic adrenal insufficiency), herpes gestationis, linear IgA bullous dermatosis, epidermolysis bullosa acquisita, alopecia areata, vitiligo, Harada disease disease), autoimmune optic neuropathy, idiopathic azoospermia, recurrent fetal loss (e.g., recurrent miscarriage or miscarriage), infertility related to lack of feto-maternal tolerance, or respiratory diseases such as asthma and ARDS (acute respiratory distress syndrome).

[0065] In some embodiments, the disease or condition is psoriasis, psoriatic arthritis, rheumatoid arthritis, contact hypersensitivity, allergic diseases, eosinophilia, eosinophilic esophagitis, respiratory diseases such as asthma and ARDS (acute respiratory distress syndrome), giant cell arteritis, polymyalgia rheumatica, primary Sjögren's syndrome, TNF-refractory rheumatoid arthritis, alopecia areata, primary biliary cholangitis (PBC), graft-versus-host disease (GvHD), vitiligo, ANCA vasculitis, type 1 diabetes, inflammatory bowel disease including Crohn's disease and ulcerative colitis, asthma, chronic periodontitis, recurrent fetal loss, celiac disease, transplant rejection, or non-infectious uveitis.

[0066] "Immune response" may require, for example, antibody production and / or immune effector cell (e.g., T cell) activation, inflammatory cytokine production, or any indication or condition described herein or otherwise known in the art. As used herein, the terms "treatment (treatment)", "treating (treating)" etc. refer to obtaining desired pharmacological and / or physiological effects. Preferably, the effect is therapeutic, that is, the effect partially or completely reduces the adverse symptoms associated with IL-2 signal conduction or the disease associated therewith. For this reason, the inventive method includes administering a "therapeutically effective amount" of a CD122 binding agent. "Therapeutically effective amount" refers to the amount of the desired therapeutic outcome effectively achieved at the desired dosage and for the desired time period. The therapeutically effective amount can vary according to factors such as individual disease state, age, sex, and weight, and the ability of the CD122 binding agent to induce a desired response in an individual. Pharmacological and / or physiological effects can be preventive, that is, the effect completely or partially prevents adverse symptoms or diseases associated with IL-2 signal conduction.

[0067] The CD122 binding agent can be a part of a composition suitable for administration to a mammal. Preferably, the composition is a pharmaceutically acceptable (e.g., physiologically acceptable) composition comprising a carrier, preferably a pharmaceutically acceptable (e.g., physiologically acceptable) carrier and amino acid sequence of the present invention, antigen binding agent or carrier. Any suitable carrier can be used within the context of the present invention, and such carriers are well known in the art. The selection of carrier will be determined in part by the specific site where the composition can be administered and the specific method for administering the composition. The composition can also include any other excipients for the preparation (particularly parenteral preparations) of therapeutic molecules (e.g., proteins or antibodies), including, for example, buffers, tension regulators, stabilizers, surfactants, etc. The composition can optionally be sterile. The composition can be frozen or lyophilized for storage and reconstituted in a suitable sterile carrier before use. The compositions can be prepared according to conventional techniques, for example, as described in Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins, Philadelphia, PA (2001).

[0068] The dosage used will depend on the specific situation in which the binding agent is used. Typical dosages of CD122 binding agents can be, for example, in the range of 1 pg / kg to 100 mg / kg of animal or human body weight; however, dosages below or above this exemplary range are also within the scope of the present invention. The therapeutic or preventive efficacy can be monitored by regular assessment of the treated patient. For repeated administration over several days or longer, depending on the condition, the treatment can be repeated until the desired suppression of disease symptoms occurs. However, other dosage regimens may be useful and within the scope of the present invention. The desired dose can be delivered by administering the composition as a single bolus, by administering the composition as multiple boluses, or by administering the composition as a continuous infusion.

[0069] The CD122 binding agent can be administered to a mammal using standard administration techniques, including oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual or suppository administration. The composition is preferably suitable for parenteral administration. As used herein, the term "parenteral" includes intravenous, intramuscular, subcutaneous, rectal, vaginal and intraperitoneal administration. In some embodiments, the composition is administered to a mammal using peripheral systemic delivery by intravenous, intraperitoneal or subcutaneous injection.

[0070] After being applied to mammals (for example, people), the bioactivity of the CD122 binding agents of the present invention can be measured by any suitable method known in the art.For example, bioactivity can be assessed by determining the stability of specific CD122 binding agents. In one embodiment of the invention, the in vivo half-life of CD122 binding agents (for example, antibodies) is between about 30 minutes and 45 days (for example, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 10 hours, about 12 hours, about 1 day, about 5 days, about 10 days, about 15 days, about 25 days, about 35 days, about 40 days, about 45 days or the scope defined by any two values ​​in the aforementioned values). In another embodiment, the in vivo half-life of the CD122 binding agent is between about 2 hours and 20 days (e.g., about 5 hours, about 10 hours, about 15 hours, about 20 hours, about 2 days, about 3 days, about 7 days, about 12 days, about 14 days, about 17 days, about 19 days, or a range defined by any two of the foregoing values). In another embodiment, the in vivo half-life of the CD122 binding agent is between about 10 days and about 40 days (e.g., about 10 days, about 13 days, about 16 days, about 18 days, about 20 days, about 23 days, about 26 days, about 29 days, about 30 days, about 33 days, about 37 days, about 38 days, about 39 days, about 40 days, or a range defined by any two of the foregoing values). In other embodiments, the biological activity of the CD122 binding agent of the present invention can be assessed by monitoring receptor occupancy on cells expressing CD122 in peripheral blood and tissues (such as skin tissue). In other embodiments, the biological activity of the CD122 binding agents of the invention can be assessed by monitoring the reduction of immune cells expressing CD122 (such as NK cells).

[0071] The CD122 binding agents of the present invention can be administered alone or in combination with other active agents or drugs. For example, CD122 binding agents can be administered in combination with other agents to treat or prevent diseases disclosed herein. In this regard, CD122 binding agents can be used in combination with at least one other inflammatory disorder or autoimmune disorder inhibitor, disease killer virus, gene therapy, cytokine therapy and adoptive T cell transfer and / or surgery, and the inflammatory disorder or autoimmune disorder inhibitor include, for example, other monoclonal antibodies. In some embodiments, CD122 binding agents and CD28 antagonists, such as cytotoxic T lymphocyte-associated protein 4 (CTLA4) (for example, CTLA4-Ig, belatacept (belatacept) or abatacept (abatacept)) are combined with (for example, conjoint therapy; administered simultaneously or sequentially in any order, or administered with any dosing regimen). The CD122 binding agents of the present invention described herein can also be used in combination with at least one other immunosuppressant, including, for example, methotrexate, corticosteroids, and other small molecule agents for treating autoimmune diseases and inflammatory diseases. In the treatment of infectious diseases by the methods of the present invention, the CD122 binding agent can be administered in combination with at least one antibacterial agent or at least one antiviral agent. In this regard, the antibacterial agent can be any suitable antibiotic known in the art. The antiviral agent can be any vaccine of any suitable type that specifically targets a particular virus (e.g., live attenuated vaccines, subunit vaccines, recombinant vector vaccines, and small molecule antiviral therapies (e.g., viral replication inhibitors and nucleoside analogs). In some embodiments, a CD122-binding agent is administered in combination (e.g., combination therapy; administered simultaneously or sequentially in any order, or with any dosing regimen) with a Janus kinase (JAK) inhibitor (e.g., abrocitinib, baricitinib, delgocitinib, fedratinib, filgotinib, oclacitinib, peficitinib, pacritinib, ruxolitinib, tofacitinib, or upadacitinib).

[0072] In addition to therapeutic uses, the CD122 binding agents described herein can also be used for diagnosis or research applications. In this regard, the CD122 binding agent can be used in a method for diagnosing cancer or infectious diseases. In a similar manner, the CD122 binding agent can be used to monitor the CD122 protein level in a subject tested for a disease or condition associated with abnormal CD122 expression. Research applications include, for example, methods using CD122 binding agents and labels to detect CD122 protein in a sample (e.g., in human body fluids or cell or tissue extracts). The CD122 binding agent can be used with or without modification (e.g., covalent or non-covalent labeling with a detectable portion). For example, the detectable portion can be a radioactive isotope (e.g., 3 H. 14 C. 32 P. 35 S or 125 I), fluorescent or chemiluminescent compounds (e.g., fluorescein isothiocyanate, rhodamine, or luciferin), enzymes (e.g., alkaline phosphatase, β-galactosidase, or horseradish peroxidase), or prosthetic groups. Any method known in the art for coupling an antigen-binding agent (e.g., an antibody) to a detectable moiety can be used in the context of the present invention (see, e.g., Hunter et al., Nature, 194:495-496 (1962); David et al., Biochemistry, 13:1014-1021 (1974); Pain et al., J. Immunol. Meth., 40:219-230 (1981); and Nygren, J. Histochem. Cytochem., 30:407-412 (1982)).

[0073] CD122 binding agents of the present invention can be used to measure CD122 protein levels by any suitable method known in the art. Such methods include, for example, radioimmunoassay (RIA) and FACS. Any suitable technology can be used to determine the normal or standard expression value of CD122 protein, for example, by combining a sample comprising or suspected of comprising CD122 polypeptides with CD122-specific antibodies under conditions suitable for forming an antigen-antibody complex. Directly or indirectly, antibodies are labeled with detectable substances to detect bound or unbound antibodies. Suitable detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, and radioactive materials (see, for example, Zola, " monoclonal antibodies: Technical Manual (MonoclonalAntibodies:A Manual of Techniques) ", CRC Publishing Company (CRC Press, Inc. (1987)). The amount of the PD-1 polypeptides expressed in the sample is then compared with a standard value.

[0074] The CD122 binding agent can be provided in a test kit, i.e., a packaged combination of a predetermined amount of reagent and instructions for performing a diagnostic assay. If the CD122 binding agent is labeled with an enzyme, the test kit includes substrates and cofactors required for the enzyme (e.g., substrate precursors providing a detectable chromophore or fluorophore). In addition, other additives such as stabilizers, buffers (e.g., blocking buffers or lysis buffers) and the like may be included in the test kit. The relative amounts of various reagents can be varied to significantly optimize the sensitivity of the assay in the concentration in the reagent solution. The reagent can be provided in the form of a dry powder (usually lyophilized), which includes an excipient that will provide a reagent solution with an appropriate concentration when dissolved.

[0075] Nucleic acid, cell, production method

[0076] The present invention also provides one or more separated or purified nucleic acid sequences, which encode CD122 binding agents or their independent heavy chain or light chain immunoglobulin polypeptides.Thus, in one embodiment, nucleic acid encoding is as provided herein immunoglobulin light chain variable region or complete immunoglobulin light chain.In another embodiment, nucleic acid encoding is as provided herein immunoglobulin heavy chain variable region or complete immunoglobulin heavy chain.In another embodiment again, nucleic acid encoding is as provided herein immunoglobulin light chain variable region or complete immunoglobulin light chain and immunoglobulin heavy chain variable region or complete immunoglobulin heavy chain.

[0077] The terms "nucleic acid" and "nucleic acid sequence" are intended to encompass polymers of DNA or RNA, i.e., polynucleotides, which can be single-stranded or double-stranded and can contain non-natural or altered nucleotides. As used herein, the terms "nucleic acid" and "polynucleotide" refer to nucleotides in a polymeric form of any length, i.e., ribonucleotides (RNA) or deoxyribonucleotides (DNA). These terms refer to the primary structure of a molecule, and thus include double-stranded and single-stranded DNA and double-stranded and single-stranded RNA. As equivalents, the terms include analogs of RNA or DNA made from nucleotide analogs and modified polynucleotides, such as, but not limited to, methylated and / or capped polynucleotides. Nucleic acids are typically connected by phosphate bonds to form nucleic acid sequences or polynucleotides, even though many other bonds (e.g., phosphorothioates, boranophosphates, etc.) are known in the art.

[0078] Nucleic acid can be part of a vector. A vector can be, for example, a plasmid, an episome, a cosmid, a viral vector (e.g., a retrovirus or adenovirus) or a phage. Suitable vectors and vector preparation methods are well known in the art (see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Press, Cold Spring Harbor, NY (2001); and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, New York, NY (1994)).

[0079] In addition to the nucleic acid sequence encoding the immunoglobulin heavy chain and / or light chain, the vector can also include expression control sequences, such as promoters, enhancers, polyadenylation signals, transcription terminators, internal ribosome entry sites (IRES), etc., for expression of the coding sequence in the host cell. Exemplary expression control sequences are well known in the art and are described in, for example, Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif., (1990).

[0080] A large number of promoters from a variety of different sources, including constitutive promoters, inducible promoters and repressible promoters are well known in the art. Representative sources of promoters include, for example, viruses, mammals, insects, plants, yeast and bacteria, and suitable promoters from these sources are easily available, or can be prepared synthetically based on sequences publicly available from depository institutions such as ATCC and other commercial or personal sources. Promoters can be unidirectional (i.e., start transcription in one direction) or bidirectional (i.e., start transcription in 3' or 5' directions). Non-limiting examples of promoters include, for example, T7 bacterial expression system, pBAD (araA) bacterial expression system, cytomegalovirus (CMV) promoter, SV40 promoter, RSV promoter. Inducible promoters include, for example, Tet system (USP 5,464,758 and 5,814,618), ecdysone inducible system (No et al., Proceedings of the National Academy of Sciences of the United States of America, 93:3346-3351 (1996)); T-REX TM System (Invitrogen Carlsbad, CA), LACSWITCH TM system (Stratagene, San Diego, CA) and the Cre-ERT tamoxifen-inducible recombinase system (Indra et al., Nucleic Acids Res., 27:4324-4327 (1999); Nucleic Acids Res., 28:e99 (2000); U.S. Patent 7,112,715; and Kramer and Fussenegger, Methods Mol. Biol., 308:123-144 (2005)).

[0081] As used herein, the term "enhancer" refers to a DNA sequence that increases, for example, the transcription of a nucleic acid sequence to which it is operably connected. The enhancer can be positioned many kilobases away from the nucleic acid sequence coding region and can mediate the combination of regulatory factors, the pattern of DNA methylation, or the variation of DNA structure. A large number of enhancers from a variety of different sources are well known in the art and can be used as cloned polynucleotides or obtained in cloned polynucleotides (for example, from depository institutions such as ATCC and other commercial or individual sources). Many polynucleotides comprising promoters (such as conventional CMV promoters) also comprise enhancer sequences. The enhancer can be located upstream, inside, or downstream of the coding sequence.

[0082] The vector may also include a selectable marker gene. As used herein, the term "selectable marker gene" refers to a nucleic acid sequence that allows cells expressing a nucleic acid sequence to be specifically selected or targeted in the presence of a corresponding selection agent. Suitable selectable marker genes are known in the art and are described, for example, in International Patent Application Publications WO 1992 / 008796 and WO 1994 / 028143; Wigler et al., Proceedings of the National Academy of Sciences of the United States of America, 77:3567-3570 (1980); O'Hare et al., Proceedings of the National Academy of Sciences of the United States of America, 78:1527-1531 (1981); Mulligan & Berg, Proceedings of the National Academy of Sciences of the United States of America, 78:2072-2076 (1981); Colberre-Garapin et al., Journal of Molecular Biology, 150:1-14 (1981); Santerre et al., Gene , 30:147-156 (1984); Kent et al., Science, 237:901-903 (1987); Wigler et al., Cell, 11:223-232 (1977); Szybalska & Szybalski, Proceedings of the National Academy of Sciences of the United States of America, 48:2026-2034 (1962); Lowy et al., Cell, 22:817-823 (1980); and U.S. Patents 5,122,464 and 5,770,359.

[0083] In some embodiments, the vector is an "episomal expression vector" or "episome," which is capable of replicating in a host cell and, in the presence of appropriate selective pressure, persists as an extrachromosomal DNA fragment within the host cell (see, e.g., Conese et al., Gene Therapy, 11: 1735-1742 (2004)). Representative commercially available episomal expression vectors include, but are not limited to, episomal plasmids utilizing the Epstein-Barr nuclear antigen 1 (EBNA1) and the Epstein-Barr virus (EBV) origin of replication (oriP). Vectors pREP4, pCEP4, pREP7, and pcDNA3.1 from Invitrogen (Carlsbad, California) and vector pBK-CMV from Stratagene (La Jolla, CA) represent non-limiting examples of episomal vectors utilizing the T antigen and SV40 origin of replication instead of EBNA1 and oriP.

[0084] Other suitable vectors include integrating expression vectors that can be randomly integrated into the DNA of the host cell, or can include recombination sites to enable specific recombination between the expression vector and the chromosome of the host cell. Such integrating expression vectors can utilize the endogenous expression control sequences of the chromosome of the host cell to achieve expression of the desired protein. Examples of vectors that integrate in a site-specific manner include, for example, the flp-in system (e.g., pcDNA3. TM 5 / FRT) or as a component of the cre-lox system found in the pExchange-6 core vector from Stratagene (La Jolla, California). Examples of vectors that randomly integrate into the host cell chromosome include, for example, pcDNA3.3 from ThermoFisher (Carlsbad, California) (when introduced in the absence of T-antigen), UCOE from Millipore (Billerica, MA), and pCI or pFN10A (ACT) FLEXI from Promega (Madison, WI). TM .

[0085] Viral vectors can also be used. Representative commercially available viral expression vectors include, but are not limited to, the adenovirus-based Per.C6 system available from Crucell, Inc. (Leiden, The Netherlands), the lentiviral-based pLP1 available from Thermo Fisher Scientific (Carlsbad, California), and the retroviral vector pFB-ERV plus pCFB-EGSH available from Agilent (Stratagene, La Jolla, California).

[0086] The nucleic acid sequence encoding the amino acid sequence of the present invention can be provided to cells on the same vector (i.e., cis). Unidirectional promoters can be used to control the expression of each nucleic acid sequence. In another embodiment, a combination of a bidirectional promoter and a unidirectional promoter can be used to control the expression of multiple nucleic acid sequences. The nucleic acid sequence encoding the amino acid sequence of the present invention can alternatively be provided to cell colonies on a separate vector (i.e., trans). Each nucleic acid sequence in each separate vector can comprise identical or different expression control sequences. Separate vectors can be provided to cells simultaneously.

[0087] A vector comprising a nucleic acid encoding an amino acid sequence of the present invention can be introduced into a host cell capable of expressing the polypeptide encoded thereby, including any suitable prokaryotic or eukaryotic cell. Thus, the present invention provides in vitro cells or cell lines comprising a vector of the present invention. The present invention also provides in vitro cells or cell lines expressing immunoglobulin heavy chain polypeptides and / or light chain polypeptides or expressing PD-1 binding agents. Preferred host cells are those that can be easily and reliably grown, have a relatively fast growth rate, have a well-characterized expression system, and can be easily and efficiently transformed or transfected.

[0088] Examples of suitable prokaryotic cells include, but are not limited to, cells from the genera Bacillus (e.g., Bacillus subtilis and Bacillus brevis), Escherichia (e.g., E. coli), Pseudomonas, Streptomyces, Salmonella, and Erwinia. Particularly useful prokaryotic cells include various strains of E. coli (e.g., K12, HB101 (ATCC No. 33694), DH5α, DH10, MC1061 (ATCC No. 53338), and CC102).

[0089] In some embodiments, the vector is introduced into a eukaryotic cell. Suitable eukaryotic cells are known in the art and include, for example, yeast cells, insect cells, and mammalian cells. Examples of suitable yeast cells include yeast cells from Kluyveromyces, Pichia, Rhino-sporidium, Saccharomyces, and Schizosaccharomyces. Preferred yeast cells include, for example, Saccharomyces cerivisae and Pichia pastoris.

[0090] Suitable insect cells are described, for example, in Kitts et al., Biotechniques, 14:810-817 (1993); Lucklow, Curr. Opin. Biotechnol., 4:564-572 (1993); and Lucklow et al., J. Virol., 67:4566-4579 (1993). Preferred insect cells include Sf-9 and HI5 (Invitrogen, Carlsbad, CA).

[0091] In some embodiments, mammalian cells are used in the present invention.Many suitable mammalian host cells are known in the art, and many mammalian host cells can be obtained from American Type Culture Collection (ATCC, Manassas, VA).The example of suitable mammalian cells includes but is not limited to Chinese hamster ovary cells (CHO) (for example, CHO-K1 ATCC No. CCL61), CHO DHFR cells (for example, Urlaub et al., Proceedings of the National Academy of Sciences of the United States, 97:4216-4220 (1980)), human embryonic kidney (HEK) 293 or 293T cells (for example, ATCC No. CRL1573) and 3T3 cells (for example, ATCC No. CCL92).Other suitable mammalian cell lines are monkey COS-1 (for example, ATCC No. CRL1650) and COS-7 cell line (for example, ATCC No. CRL1651) and CV-1 cell line (for example, ATCC No. CCL70). Other exemplary mammalian host cells include primate cell lines and rodent cell lines, including mouse cell line NS0, i.e., derivatives of mouse myeloma cell line MOPC21 (e.g., Tysabri), and transformed cell lines. Normal diploid cells, cell strains derived from in vitro cultures of primary tissues, and primary explants are also suitable. Other suitable mammalian cell lines include, but are not limited to, mouse neuroblastoma N2A cells, HeLa, mouse L-929 cells, and BHK or HaK hamster cell lines, all of which are available from ATCC. Methods for selecting suitable mammalian host cells and methods for cell transformation, cultivation, amplification, screening, and purification are known in the art.

[0092] In some embodiments, the mammalian cell is a human cell. For example, the mammalian cell can be a human lymphoid cell line or a lymphoid-derived cell line, such as a cell line derived from a pre-B lymphocyte. Examples of human lymphoid cell lines include, but are not limited to, RAMOS (e.g., CRL-1596), Daudi (e.g., CCL-213), EB-3 (e.g., CCL-85), Raji cells (e.g., CCL-86), and derivatives thereof.

[0093] The nucleic acid sequence encoding the amino acid sequence of the present invention can be introduced into the cell by any suitable technique, such as by "transfection", "conversion" or "transduction". As used herein, "transfection", "conversion" or "transduction" refers to the introduction of one or more exogenous polynucleotides into a host cell using physical or chemical methods. Many transfection techniques are known in the art and include, for example, calcium phosphate DNA coprecipitation (see, for example, Murray EJ (ed.), Methods in Molecular Biology, Vol. 7, Gene Transfer and Expression Protocols, Humana Press (1991)); DEAE-dextran; electroporation; cationic liposome-mediated transfection; tungsten particles promote microparticle bombardment (Johnston, Nature, 346: 776-777 (1990)); and strontium phosphate DNA coprecipitation (Brash et al., Mol. Cell Biol., 7: 2031-2034 (1987)). Phage or viral vectors can be introduced into host cells following growth of infectious particles in suitable packaging cells, many of which are commercially available.

[0094] Nucleic acids and cells can be used for any purpose, such as for preparing the CD122 binding agents described herein. In this regard, the invention provides a method for preparing a CD122 binding agent, comprising culturing cells comprising nucleic acids encoding heavy and / or light immunoglobulin polypeptides of CD122 binding agents. In other words, the method comprises expressing nucleic acids encoding heavy and / or light chains of immunoglobulins of CD122 binding agents in cells. It should be understood that heavy and light chains of immunoglobulins can be expressed by a single nucleic acid in a given cell, or heavy and light chains of immunoglobulins can be expressed by separate nucleic acids in the same cell. The method may further comprise collecting and / or purifying the CD122 binding agent from cells or cell culture medium using known techniques.

[0095] The following examples further illustrate the invention but, of course, should not be construed as limiting its scope in any way.

[0096] Example

[0097] The following examples describe specific CD122 binding agent heavy chain polypeptides and light chain polypeptide sequences according to embodiments of the present invention. The antibodies used in these examples are as follows.

[0098] The antibodies were produced during transient expression in ExpiCHO-S mammalian cells and purified by protein A affinity chromatography. The antibody sequences of the M10-1275 and M6-p1625 series of antibodies are summarized in Tables 1A and 1B, respectively, where "H" chain and "L" chain refer to the heavy chain and light chain. The CDRs of these antibodies, as determined according to Kabat numbering, are presented in Tables 1C and 1D. The amino acid sequences and nucleic acid sequences of the complete heavy and light chains of certain antibodies are summarized in Table 1E. M10-1275 and M6-p1625 represent two classes of antibodies whose members were developed by mutagenesis and screening from parental heavy and light chains.

[0099] Table 1A

[0100] M10-1275 H chain variable region L chain variable region APE15446 SEQ ID NO:24 SEQ ID NO:35 APE15276 SEQ ID NO:22 SEQ ID NO:35 APE15173 SEQ ID NO:16 SEQ ID NO:35 APE15172 SEQ ID NO:16 SEQ ID NO:32 APE15171 SEQ ID NO:16 SEQ ID NO:36 APE15170 SEQ ID NO:23 SEQ ID NO:35 APE15169 SEQ ID NO:23 SEQ ID NO:32 APE15168 SEQ ID NO:23 SEQ ID NO:36 APE15167 SEQ ID NO: 19 SEQ ID NO:28 APE15145 SEQ ID NO: 17 SEQ ID NO:28 APE15144 SEQ ID NO: 18 SEQ ID NO:28 APE15143 SEQ ID NO:15 SEQ ID NO:28 APE15142 SEQ ID NO:16 SEQ ID NO:28 APE15141 SEQ ID NO:20 SEQ ID NO:28 APE15140 SEQ ID NO:23 SEQ ID NO:28 APE15139 SEQ ID NO:21 SEQ ID NO:28 APE15138 SEQ ID NO:12 SEQ ID NO:28 APE15137 SEQ ID NO:13 SEQ ID NO:28 APE15136 SEQ ID NO:14 SEQ ID NO:28 APE15135 SEQ ID NO:11 SEQ ID NO:28 APE15059 SEQ ID NO:2 SEQ ID NO:28 APE15058 SEQ ID NO:2 SEQ ID NO:30 APE15057 SEQ ID NO:2 SEQ ID NO:27 APE15056 SEQ ID NO:2 SEQ ID NO:35 APE15055 SEQ ID NO:2 SEQ ID NO:34 APE15054 SEQ ID NO:2 SEQ ID NO:33 APE15053 SEQ ID NO:2 SEQ ID NO:32 APE15038 SEQ ID NO: 10 SEQ ID NO:31 APE15037 SEQ ID NO:9 SEQ ID NO:31 APE15036 SEQ ID NO:8 SEQ ID NO:31 APE15035 SEQ ID NO:4 SEQ ID NO:31 APE15034 SEQ ID NO:3 SEQ ID NO:31 APE15033 SEQ ID NO:5 SEQ ID NO:31 APE15032 SEQ ID NO:7 SEQ ID NO:31 APE15031 SEQ ID NO:6 SEQ ID NO:31 APE14970 SEQ ID NO:2 SEQ ID NO:31 APE14969 SEQ ID NO:2 SEQ ID NO:29 APE14971 SEQ ID NO:2 SEQ ID NO:26 APE12705 SEQ ID NO:1 SEQ ID NO:25

[0101] Table 1B

[0102]

[0103]

[0104] Table 1C

[0105]

[0106]

[0107] Table 1D

[0108]

[0109]

[0110] Table 1E

[0111]

[0112] Example 1

[0113] This example demonstrates the binding kinetics (affinity) of the antibodies disclosed herein.

[0114] Binding affinity was determined using surface plasmon resonance (SPR). M6-1625 and M10-1275 antibodies were captured onto a Cytiva Protein A Chip at 0.5 μg / mL for 60 seconds. Antibodies were exposed to recombinant human IL-2Rβ or cynomolgus monkey IL-2Rβ (corresponding to residues Ala1-Thr215 of the extracellular domain, excluding the signal peptide) at typical concentrations ranging from 0.4 nM to 11 nM using a Biacore S200 instrument with typical association of 300 seconds and dissociation of 1200 seconds. Sensorgrams were globally fitted using a 1:1 binding model in the Biacore T200 evaluation software to calculate association and dissociation rates as well as K D The K values ​​of the CD122 binding agents disclosed herein obtained by SPR are shown in Table 1. D A summary of the measurement results is shown in Tables 2A and 2B.

[0115] Table 2A: M6-p1625 binding affinity data

[0116]

[0117]

[0118] Table 2B: M10-1275 binding affinity data

[0119] Antibody <![CDATA[Human IL-2RβK D (SPR)]]> <![CDATA[Cynomolgus monkey IL-2RβK D (SPR) <!-- 25 -->]]> APE15446 10.7 pM 4.15 pM APE15276 17.8 pM 5.99 pM APE15173 18.4 pM 5.09 pM APE15172 116 pM 27.1 pM APE15171 276 pM 52.2 pM APE15170 12.8 pM 4.87 pM APE15169 99.5 pM 14.4 pM APE15168 324 pM 37.9 pM APE15167 45 pM 6.55 pM APE15145 29.6 pM 10.8 pM APE15144 29.7 pM 11.6 pM APE15143 25.1 pM 9.72 pM APE15142 22.2 pM 10.5 pM APE15141 22.0 pM 10.3 pM APE15140 21.7 pM 10.2 pM APE15139 35.2 pM 9.23 pM APE15138 43.6 pM 16.3 pM APE15137 51.8 pM 17.4 pM APE15136 48.1 pM 11.4 pM APE15135 28.2 pM 10.5 pM APE15059 69.7 pM 15.9 pM APE15058 154 pM 22.3 pM APE15057 144 pM 24 pM APE15056 122 pM 19.8 pM APE15055 369 pM 83.8 pM ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ APE14969 187pM 18pM APE14971 278pM 26pM APE12705 80pM 20pM

[0120] Example 2

[0121] This example shows that the anti-CD122 chimeric antibody of the present invention binds to human or cynomolgus monkey IL-2Rβγ expressed on CHO-K1 cells in a dose-dependent manner.

[0122] CHO-K1 cells stably expressing full-length human or cynomolgus monkey IL-2Rβ and IL-2Rγ in the logarithmic growth phase were harvested with Accutase solution, resuspended in FACS buffer, and plated at 100,000 cells / well in a 96-well plate. Figure 1 and 2 Cells were incubated with the indicated concentrations of antibodies at 4°C on a plate shaker for 45 minutes. After washing, cells were incubated with a secondary goat anti-human κ DyL650 antibody for 20 minutes on a plate shaker at 4°C. Cells were washed again and analyzed on a FACSArray flow cytometer. Data were analyzed using Novoexpress and GraphPad Prism software. Figure 1 and 2The results are shown in Table 3. Table 3 lists the binding properties of anti-CD122 chimeric antibodies: affinity (K) for human and cynomolgus monkey CD122 proteins D , measured by SPR assay), the ability to block IL-2 (SPR assay), and the EC50 binding to cell-expressed human and cynomolgus monkey CD122 / CD132 (cell-based receptor binding assay).

[0123] Table 3

[0124]

[0125] Example 3

[0126] This example demonstrates that the anti-CD122 antibodies of the present invention can inhibit IL-2-induced or IL-15-induced pSTAT5 luciferase activity in HEK cells expressing human IL-2βγ.

[0127] HEK293 cells stably transfected with IL-2Rβ, IL-2Rγ and STAT5 luciferase reporter genes were plated at 50,000 cells per well in 96-well plates. Chimeric antibodies or JAK inhibitors were added as described in the previous section. Figure 3 The concentration indicated in (A) starts titration with 100nM and is added to the cell and continues within 30 minutes.Then recombinant IL-2 or IL-15 is added to the cell and continues another 3.5 hours.Add SteadyGlo substrate, continue 10 minutes, and read plate on GloMax machine.Use GraphPadPrism software that data is analyzed. Figure 3 The results are shown in .

[0128] Example 4

[0129] This example demonstrates that anti-CD122 IgG4 antibodies can inhibit IL-2 or IL-15-induced proliferation of PHA-stimulated primary human T cell blasts.

[0130] Primary human PBMCs were isolated from whole blood donors and stimulated with 5 μg / mL PHA for 3 days in culture in T75 flasks. Cells were washed, counted, and plated at 100,000 cells / well in 96-well plates. Antibodies were added as described. Figure 4 The concentration indicated in (A) was titrated at a 1:3 dilution starting at 100 nM and added to the cells. Recombinant IL-2 (15 pM) or IL-15 (150 pM) was added to the cells. The cells were cultured for another three days. SteadyGlo substrate was added, continued for 10 minutes, and the plate was read on a GloMax machine. Graphpad Prism software was used to analyze the data.

[0131] Figure 4The results are shown in . Both M10-1275 and M6-1625 were superior to upadacitinib in blocking IL-2 and IL-15.

[0132] Example 5

[0133] This example shows that anti-CD122 IgG4 antibodies bind to human IL-2Rβγ expressed on CHO-K1 cells in a dose-dependent manner. CHO-K1 cells stably expressing full-length human or cynomolgus monkey IL-2Rβ and IL-2Rγ were harvested in the logarithmic growth phase with Accutase solution, resuspended in FACS buffer, and plated at 100,000 cells / well in 96-well plates. Figure 5 Cells were incubated with the indicated concentrations of antibodies at 4°C on a plate shaker for 45 minutes. After washing, cells were incubated with a secondary goat anti-human κ DyL650 antibody for 20 minutes on a plate shaker at 4°C. Cells were washed again and analyzed on a FACSArray flow cytometer. Data were analyzed using Novoexpress and GraphPad Prism software.

[0134] Figure 5 The results are shown in. All antibodies showed human and cynomolgus monkey binding with EC50 in the range of 0.3-0.1 nM.

[0135] Example 6

[0136] This example demonstrates that humanized anti-CD122 IgG4 antibodies inhibit IL-2 or IL-15-induced proliferation of purified primary human NK cells.

[0137] Primary human PBMCs were isolated from whole blood donors and untouched NK cells were isolated using a Miltenyi bead kit. 100,000 cells were plated on a 96-well plate. Antibodies were titrated at a 1:10 dilution starting at 100 nM at the indicated concentrations and added to the cells. Recombinant IL-2 (50 pM) or recombinant IL-15 (60 pM) was added to the cells. The cells were cultured for 5 days. Finally, SteadyGlo substrate was added for 10 minutes and the plate was read on a GloMax machine. Data were analyzed using Graphpad Prism software. Figure 6 The results are shown in .

[0138] Example 7

[0139] This example demonstrates that humanized anti-CD122 IgG4 antibodies inhibit IL-2 or IL-15-induced proliferation and survival of primary human NK cells gated within PBMC cultures.

[0140] Primary human PBMCs were isolated from whole blood donors, labeled with CFSE, and plated at 250,000 cells / well in 96-well plates. Figure 7 The concentration indicated in the titration was started at 100 nM and diluted 1:3 and added to the cells. Recombinant IL-2 (250 pM) or recombinant IL-15 (200 pM) was added to the cells. The cells were incubated for 5 days. The cells were stained with CD56 and CD3 to gate the NK cell population and stained with Annexin V as a measure of apoptotic cells. The cells were analyzed on a flow cytometer and the data were analyzed using Novoexpress and GraphPad Prism software. Proliferation was expressed as a % relative to cells with a diluted CFSE peak (beyond the unstimulated fraction), and apoptosis was indicated by Annexin V staining, as shown in Table 2. Figure 7 Both antibodies tested effectively inhibited IL-2 or IL-15 induced proliferation as measured by CFSE.

[0141] Example 8

[0142] This example demonstrates that humanized anti-CD122 IgG4 antibodies inhibit the binding of IL-2 or IL-15 to HEK cells expressing human IL-2βγ.

[0143] HEK293 cells stably transfected with human IL-2Rβ and IL-2Rγ were plated at 100,000 cells / well in 96-well plates. Figure 8 The concentration indicated in (A) starts with 100nM and is diluted 1:3 to titrate, and is added to the cells. Recombinant IL-2 (5nM) or recombinant IL-15 (2nM) are also added to the cells. The cells are incubated on a plate shaker at 4°C for 30 minutes. The cells are washed and then analyzed on a flow cytometer, and Novoexpress and GraphPadPrism software are used to analyze the data. Figure 8 The results are shown in .

[0144] Example 9

[0145] This example demonstrates that humanized anti-CD122 IgG4 antibodies inhibit IL-15 or IL-15 / IL1-5RA-induced proliferation of primary human T cell blasts.

[0146] Primary human PBMCs were isolated from whole blood donors and stimulated with 5 μg / mL PHA in culture for 3 days in T75 flasks. Cells were washed, counted, and plated at 100,000 cells / well in 96-well plates. Antibodies were added as described. Figure 9 The concentrations indicated in the table were titrated starting at 100 nM at a 1:3 dilution and added to the cells. Recombinant IL-15 alone (1 nM) or recombinant IL-15 + IL-15Ra (1 nM each) were added to the cells. The cells were cultured for an additional three days. Finally, SteadyGlo substrate was added for 10 minutes and the plate was read on a GloMax machine. Data were analyzed using Graphpad Prism software. Figure 9 The results are shown in .

[0147] Example 10

[0148] This example demonstrates that humanized anti-CD122 IgG4 antibodies inhibit IL-15 or IL-15 / IL1-5RA-induced proliferation of primary human NK cells.

[0149] Primary human PBMCs were isolated from whole blood donors, and naive NK cells were isolated using the Miltenyi beads kit. 100,000 cells were plated in 96-well plates. Antibodies were added as Figure 10 The concentrations indicated in the table were titrated starting at 100 nM at a 1:10 dilution and added to the cells. Recombinant IL-15 (60 pM) or recombinant IL-15:IL15RA (60 pM each) were added to the cells. The cells were cultured for 5 days. Finally, SteadyGlo substrate was added for 10 minutes and the plate was read on a GloMax machine. Data were analyzed using Graphpad Prism software. Figure 10 The results are shown in .

[0150] Example 11

[0151] This example demonstrates that humanized anti-CD122 IgG1 antibody inhibits IL-2 or IL-15-induced proliferation of primary human NK cells.

[0152] Primary human PBMCs were isolated from whole blood donors, and naive NK cells were isolated using the Miltenyi beads kit. 100,000 cells were plated in 96-well plates. Antibodies were added as Figure 11The concentration indicated in (A) was titrated at a 1:10 dilution starting at 100 nM and added to the cells. Recombinant IL-2 (50 pM) or recombinant IL-15 (60 pM) was added to the cells. The cells were cultured for 5 days. Finally, SteadyGlo substrate was added, continued for 10 minutes, and the plate was read on a GloMax machine. Graphpad Prism software was used to analyze the data. Figure 11 The results are shown in .

[0153] Example 12

[0154] This example demonstrates that humanized anti-CD122 IgG1 antibody inhibits IL-2 or IL-15-induced proliferation of primary human pan T cells.

[0155] Primary human PBMCs were isolated from whole blood donors, and untouched pan T cells were isolated using the Miltenyi beads kit. 100,000 cells were plated in 96-well plates. Antibodies were added as Figure 12 The concentration indicated in (A) was titrated at a 1:3 dilution starting at 100 nM and added to the cells. Recombinant IL-2 (1 nM) or recombinant IL-15 was added to the cells. The cells were cultured for 7 days. Finally, SteadyGlo substrate was added, continued for 10 minutes, and the plate was read on a GloMax machine. Graphpad Prism software was used to analyze the data. Figure 12 The results are shown in .

[0156] Example 13

[0157] This example demonstrates that humanized anti-CD122 antibodies exhibit in vivo efficacy in a xenogeneic NSG-Tg(huIL-15) / Hu-PBMC graft-versus-host disease (GvHD) model.

[0158] A xenogeneic NSG-Tg (huIL-15) / Hu-PBMC GvHD model was developed. null(NSG) human IL-15 transgenic mice were irradiated and then injected intravenously with human PBMCs in each mouse. Antibodies (APE15170-IgG4P, APE15295-IgG4P, CTLA4-Ig (positive control) and a combination of APE15170-IgG4 and CTLA4-Ig) were administered intraperitoneally. APE15170-IgG4P, APE15295-IgG4P and APE15170-IgG4P were administered twice a week at 10 mg / kg for 4 weeks, and CTLA4-Ig was administered three times a week at 75 μg, both starting the day after PBMC injection. A control IgG4 isotype was also administered twice a week.

[0159] In terms of preventing death, the anti-CD122 antagonist antibodies disclosed herein showed significant improvement relative to isotype control and CTLA4-Ig alone. The combination of APE15170-IgG4P and CTLA4-Ig showed even greater improvement compared to either APE15170-IgG4P or APE15295-IgG4P alone. Figure 13 The results are shown in .

[0160] Example 14

[0161] This example demonstrates that humanized anti-CD122 antibodies exhibit in vivo efficacy in a xenogeneic NSG (huIL-15Tg) / Hu-PBMC graft-versus-host disease (GvHD) model.

[0162] At the Jackson Laboratory The Jackson laboratory InVivo Pharmacology Services (Sacramento, CA) performed a xenogeneic NSG (huIL-15Tg) / Hu-PBMC GvHD model to test the efficacy of the anti-CD122 antibodies disclosed herein. null (NSG) human IL-15 transgenic mice were irradiated, followed by intravenous injection of human PBMCs in each mouse. On the second day after PBMC injection, APE15428 (IgG1-LALA) was administered intraperitoneally at 10 mg / kg, 3 mg / kg, or 1 mg / kg twice a week for 4 consecutive weeks. Disease was monitored three times a week by weight loss, death, and measurement of the following GvHD scores: weight loss, activity, fur texture, pallor, and posture. Animals that showed more than 10% weight loss were monitored for disease daily, and animals that showed more than 20% weight loss relative to the starting weight were euthanized.

[0163] Anti-CD122 antagonist antibodies disclosed herein show statistically significant efficacy relative to isotype controls in terms of the median time to survival. Provided are the percentages relative to starting body weight of individual animals when isotype controls, anti-CD122 antibody APE15428 (IgG1LALA) and CTLA-4-Ig (positive control) are administered respectively. Figure 14 The results are shown in .

[0164] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0165] Unless otherwise indicated herein or clearly contradicted by context, the use of the terms "a" and "an," as well as "the" and "at least one," and similar indicators in the context of describing the present invention (especially in the context of the following claims) should be interpreted to encompass both the singular and the plural. Unless otherwise indicated herein or clearly contradicted by context, the use of the term "at least one" after a list of one or more items (e.g., "at least one of A and B") should be interpreted to mean one item (A or B) selected from the listed items or any combination of two or more items (A and B) of the listed items. Unless otherwise indicated herein, the terms "comprising," "having," "including," and "containing" should be interpreted as open-ended terms (i.e., meaning "including but not limited to") unless otherwise indicated herein. Unless otherwise indicated herein, the recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each individual value falling within the range, and each individual value is incorporated into the specification as if recited individually herein. Unless otherwise noted herein or obviously contradictory to the context, all methods described herein may be performed in any suitable order. The use of any and all examples or exemplary language (e.g., "such as") provided herein is intended only to better describe the present invention and is not intended to limit the scope of the present invention, unless otherwise stated. Any language in this specification should not be construed as indicating any unclaimed element as necessary for practicing the present invention.

[0166] Preferred embodiments of the present invention are described herein, including the best modes for carrying out the present invention known to the inventors. After reading the foregoing description, variations of those preferred embodiments may become apparent to those of ordinary skill in the art. The inventors expect that technicians will adopt these variations when appropriate, and the inventors intend that the present invention be put into practice in a manner different from that specifically described herein. Therefore, where permitted by applicable law, the present invention includes all modifications and equivalent forms of the subject matter recited in the appended claims. In addition, the present invention encompasses any combination of the above-mentioned elements in all possible variations thereof, unless otherwise noted herein or otherwise clearly contradictory to the context.

Claims

1. A CD122 binding agent comprising an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region, wherein: (a) When determined according to Kabat numbering, The immunoglobulin heavy chain variable region comprises: CDR1 comprising SEQ ID NO: 108; CDR2 comprising SEQ ID NO: 109; and CDR3, the CDR3 comprising SEQ ID NO: 98; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 110; CDR2 comprising SEQ ID NO: 111; and CDR3, the CDR3 comprising SEQ ID NO: 112; wherein, optionally, when determined according to Kabat numbering: The immunoglobulin heavy chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 78; CDR2 comprising SEQ ID NO: 94; and CDR3, the CDR3 comprising SEQ ID NO: 98; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 99; CDR2 comprising SEQ ID NO: 103; and CDR3, the CDR3 comprising SEQ ID NO: 105; (b) the immunoglobulin heavy chain variable region comprises SEQ ID NO: 113, and the immunoglobulin light chain variable region comprises SEQ ID NO: 114; (c) the immunoglobulin heavy chain variable region comprises at least 80% sequence identity to any one of SEQ ID NOs: 37-69, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 37-69, and the immunoglobulin light chain variable region comprises at least 80% sequence identity to any one of SEQ ID NOs: 70-77, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 70-77; Optionally, wherein the immunoglobulin heavy chain variable region comprises at least 80% sequence identity to any one of SEQ ID NO: 68 or 69, and / or comprises at least the CDR regions of any one of SEQ ID NO: 68 or 69, and the immunoglobulin light chain variable region comprises at least 80% sequence identity to SEQ ID NO: 77, and / or comprises at least the CDR regions of SEQ ID NO: 77; (e) when determined according to Kabat numbering, the immunoglobulin heavy chain variable region comprises: CDR1, the CDR1 comprising SEQ ID NO: 146; CDR2 comprising SEQ ID NO: 147; and CDR3, the CDR3 comprising SEQ ID NO: 148; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 138; CDR2 comprising SEQ ID NO: 139; and CDR3, the CDR3 comprising SEQ ID NO: 140; wherein, optionally, when determined according to Kabat numbering: The immunoglobulin heavy chain variable region comprises: CDR1 comprising SEQ ID NO: 117 or 124; CDR2 comprising SEQ ID NO: 127; and CDR3, the CDR3 comprising SEQ ID NO: 136; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 138; CDR2 comprising SEQ ID NO: 139; and CDR3 comprising SEQ ID NO: 144 or 145; (d) the immunoglobulin heavy chain variable region comprises SEQ ID NO: 150, and the immunoglobulin light chain variable region comprises SEQ ID NO: 151; or (f) the immunoglobulin heavy chain variable region comprises at least 80% sequence identity to any one of SEQ ID NOs: 1-24, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 1-24, and the immunoglobulin light chain variable region comprises at least 80% sequence identity to SEQ ID NOs: 25-36, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 25-36; Optionally, the immunoglobulin heavy chain variable region comprises at least 80% sequence identity to any one of SEQ ID NO: 16, 23 or 24, and / or comprises at least the CDR region of any one of SEQ ID NO: 16, 23 or 24, and the immunoglobulin light chain variable region comprises at least 80% sequence identity to SEQ ID NO: 28 or 35, and / or comprises at least the CDR region of SEQ ID NO: 28 or 35.

2. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to any one of SEQ ID NOs: 37-69, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 37-69.

3. The CD122 binding agent of claim 1, wherein the immunoglobulin light chain variable region comprises at least 90% sequence identity to any one of SEQ ID NOs: 70-77, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 70-77.

4. The CD122 binding agent of any one of claims 1 to 3, wherein the immunoglobulin heavy chain variable region comprises: CDR1 comprising SEQ ID NO: 108; CDR2, comprising SEQ ID NO: 109; CDR3, the CDR3 comprising SEQ ID NO: 98; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 110; CDR2, comprising SEQ ID NO: 111; CDR3, wherein the CDR3 comprises SEQ ID NO:

112.

5. The CD122 binding agent of claim 4, wherein the immunoglobulin heavy chain variable region CDR1 comprises any one of SEQ ID NOs: 78-93.

6. The CD122 binding agent of claim 4 or 5, wherein the immunoglobulin heavy chain variable region CDR2 comprises any one of SEQ ID NOs: 94-97.

7. The CD122 binding agent of any one of claims 4 to 6, wherein the immunoglobulin light chain variable region CDR1 comprises SEQ ID NOs: 99-102.

8. The CD122 binding agent of any one of claims 4 to 7, wherein the immunoglobulin light chain variable region CDR2 comprises SEQ ID NO: 103 or 104.

9. The CD122 binding agent of any one of claims 4 to 8, wherein the immunoglobulin light chain variable region CDR3 comprises SEQ ID NO: 105-107.

10. The CD122 binding agent of any one of claims 4 to 9, comprising the immunoglobulin heavy chain variable region of any one of SEQ ID NOs: 37-69.

11. The CD122 binding agent of any one of claims 4 to 10, comprising the immunoglobulin light chain variable region of any one of SEQ ID NOs: 70-77.

12. The CD122 binding agent of claim 4, wherein the immunoglobulin heavy chain variable region comprises: CDR1, the CDR1 comprising SEQ ID NO: 78; CDR2 comprising SEQ ID NO: 94; and CDR3, the CDR3 comprising SEQ ID NO: 98; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 99; CDR2 comprising SEQ ID NO: 103; and CDR3, wherein the CDR3 comprises SEQ ID NO:

105.

13. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to SEQ ID NO: 69, or at least comprises the CDRs of SEQ ID NO: 69, and the immunoglobulin light chain variable region comprises at least 90% sequence identity to SEQ ID NO: 77, or at least comprises the CDRs of SEQ ID NO:

77.

14. The CD122 binding agent of claim 13, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 69, and the immunoglobulin light chain variable region comprises SEQ ID NO:

77.

15. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to SEQ ID NO: 68, or comprises at least the CDRs of SEQ ID NO: 68, and the immunoglobulin light chain variable region comprises at least 90% sequence identity to SEQ ID NO: 77, or comprises at least the CDRs of SEQ ID NO:

77.

16. The CD122 binding agent of claim 15, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 68, and the immunoglobulin light chain variable region comprises SEQ ID NO:

77.

17. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to any one of SEQ ID NOs: 1-24, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 1-24.

18. The CD122 binding agent of claim 1, wherein the immunoglobulin light chain variable region comprises at least 90% sequence identity to any one of SEQ ID NOs: 25-36, and / or comprises at least the CDR regions of any one of SEQ ID NOs: 25-36.

19. The CD122 binding agent of any one of claims 1, 17, or 18, wherein the immunoglobulin heavy chain variable region comprises, when determined according to Kabat numbering: CDR1, the CDR1 comprising SEQ ID NO: 146; CDR2 comprising SEQ ID NO: 147; and CDR3, the CDR3 comprising SEQ ID NO: 148; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 138; CDR2 comprising SEQ ID NO: 139; and CDR3, wherein the CDR3 comprises SEQ ID NO:

149.

20. The CD122 binding agent of claim 19, wherein the immunoglobulin heavy chain variable region CDR1 comprises any one of SEQ ID NOs: 117-126.

21. The CD122 binding agent of claim 19 or 20, wherein the immunoglobulin heavy chain variable region CDR2 comprises any one of SEQ ID NOs: 127-135.

22. The CD122 binding agent of any one of claims 19 to 21, wherein the immunoglobulin heavy chain variable region CDR3 comprises SEQ ID NO: 136 or 137.

23. The CD122 binding agent of any one of claims 19 to 22, wherein the immunoglobulin light chain variable region CDR3 comprises any one of SEQ ID NOs: 140-145.

24. The CD122 binding agent of any one of claims 19 to 23, comprising the immunoglobulin heavy chain variable region of any one of SEQ ID NOs: 1-24.

25. The CD122 binding agent of any one of claims 19 to 24, comprising the immunoglobulin light chain variable region of any one of SEQ ID NOs: 25-36.

26. The CD122 binding agent of claim 12, wherein the immunoglobulin heavy chain variable region comprises: CDR1, the CDR1 comprising SEQ ID NO: 117; CDR2 comprising SEQ ID NO: 127; and CDR3, the CDR3 comprising SEQ ID NO: 136; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 138; CDR2 comprising SEQ ID NO: 139; and CDR3, wherein the CDR3 comprises SEQ ID NO:

144.

27. The CD122 binding agent of claim 12, wherein the immunoglobulin heavy chain variable region comprises: CDR1, the CDR1 comprising SEQ ID NO: 124; CDR2 comprising SEQ ID NO: 127; and CDR3, the CDR3 comprising SEQ ID NO: 136; And the immunoglobulin light chain variable region comprises CDR1, the CDR1 comprising SEQ ID NO: 138; CDR2 comprising SEQ ID NO: 139; and CDR3, wherein the CDR3 comprises SEQ ID NO:

145.

28. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to SEQ ID NO: 24, or at least comprises the CDRs of SEQ ID NO: 24, and the immunoglobulin light chain variable region comprises at least 90% sequence identity to SEQ ID NO: 35, or at least comprises the CDRs of SEQ ID NO:

35.

29. The CD122 binding agent of claim 28, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 24, and the immunoglobulin light chain variable region comprises SEQ ID NO:

35.

30. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to SEQ ID NO: 23, or comprises at least the CDRs of SEQ ID NO: 23, and the immunoglobulin light chain variable region comprises at least 90% sequence identity to SEQ ID NO: 35, or comprises at least the CDRs of SEQ ID NO:

35.

31. The CD122 binding agent of claim 30, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 23, and the immunoglobulin light chain variable region comprises SEQ ID NO:

35.

32. The CD122 binding agent of claim 1, wherein the immunoglobulin heavy chain variable region comprises at least 90% sequence identity to SEQ ID NO: 16, or comprises at least the CDRs of SEQ ID NO: 16, and the immunoglobulin light chain variable region comprises at least 90% sequence identity to SEQ ID NO: 28, or comprises at least the CDRs of SEQ ID NO:

28.

33. The CD122 binding agent of claim 32, wherein the immunoglobulin heavy chain variable region comprises SEQ ID NO: 16, and the immunoglobulin light chain variable region comprises SEQ ID NO:

28.

34. The CD122 binding agent of any one of claims 1 to 33, wherein the CD122 binding agent is an antibody, an antigen-binding antibody fragment, or a conjugate thereof.

35. The CD122 binding agent of any one of claims 1 to 34, wherein the CD122 binding agent is F(ab')2, Fab', Fab, Fv, scFv, dsFv, or a single-chain binding polypeptide.

36. The CD122 binding agent of any one of claims 1 to 35, wherein the CD122 binding agent comprises an IgG Fc region that binds to an Fc receptor on an antigen presenting cell, optionally comprising an Fc region of IgG1.

37. A pharmaceutical composition comprising (a) a CD122 binding agent according to any one of claims 1 to 36, and (b) a pharmaceutically acceptable carrier.

38. A method of suppressing an immune response in a mammal, the method comprising administering to the mammal a CD122 binding agent according to any one of claims 1 to 36 or a pharmaceutical composition according to claim 37.

39. A method of treating an inflammatory disease or an autoimmune disease in a mammal, the method comprising administering to the mammal a CD122 binding agent according to any one of claims 1 to 36 or a pharmaceutical composition according to claim 37.

40. The method of claim 39, wherein the inflammatory disease or autoimmune disease is psoriasis, psoriatic arthritis, rheumatoid arthritis, contact hypersensitivity, allergic disease, eosinophilia, eosinophilic esophagitis, respiratory diseases such as asthma and ARDS (acute respiratory distress syndrome), giant cell arteritis, polymyalgia rheumatica, primary Sjögren's syndrome, TNF-refractory rheumatoid arthritis, alopecia areata, primary biliary cholangitis (PBC), graft-versus-host disease (GvHD), vitiligo, ANCA vasculitis, type 1 diabetes, inflammatory bowel disease including Crohn's disease and ulcerative colitis, asthma, chronic periodontitis, recurrent fetal loss, celiac disease, transplant rejection, or non-infectious uveitis.

41. The method of any one of claims 38 to 40, further comprising administering to the mammal a CD28 antagonist or a JAK kinase inhibitor.

42. A nucleic acid encoding an immunoglobulin heavy chain and / or an immunoglobulin light chain of the CD122 binding agent of any one of claims 1 to 36, optionally in a vector.

43. A cell expressing the CD122 binding agent of any one of claims 1 to 36.

44. A method for preparing a CD122 binding agent according to any one of claims 1 to 36, comprising expressing in a cell a nucleic acid sequence encoding an immunoglobulin heavy chain of the CD122 binding agent according to any one of claims 1 to 36 and a nucleic acid sequence encoding an immunoglobulin light chain of the CD122 binding agent according to any one of claims 1 to 36.

45. A CD122 binding agent according to any one of claims 1 to 36 or a pharmaceutical composition according to claim 37 for use in suppressing an immune response in a mammal.

46. ​​The CD122 binding agent of any one of claims 1 to 36 or the pharmaceutical composition of claim 37 for use in treating an inflammatory disease or an autoimmune disease in a mammal.

47. The CD122 binding agent or pharmaceutical composition of claim 46, wherein the inflammatory disease or autoimmune disease is psoriasis, psoriatic arthritis, rheumatoid arthritis, contact hypersensitivity, allergic disease, eosinophilia, eosinophilic esophagitis, respiratory diseases such as asthma and ARDS (acute respiratory distress syndrome), giant cell arteritis, polymyalgia rheumatica, primary Sjögren's syndrome, TNF-refractory rheumatoid arthritis, alopecia areata, primary biliary cholangitis (PBC), graft-versus-host disease (GvHD), vitiligo, ANCA vasculitis, type 1 diabetes, inflammatory bowel disease including Crohn's disease and ulcerative colitis, asthma, chronic periodontitis, recurrent fetal loss, celiac disease, transplant rejection, or non-infectious uveitis.

Images