JPWO2019195452A5

JPWO2019195452A5 -

Info

Publication number: JPWO2019195452A5
Application number: JP2020554172A
Authority: JP
Publication date: 2022-04-12

Claims

Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) and / or Protein
Approximately amino acid residues 21 as determined by fast photochemical oxidation (FPOP) epitope mapping
Epitope located in a discontinuous region spanning ~ 41 and 52 ~ 57 (its sequence is SEQ ID NO: 1)
A monoclonal antibody or antigen-binding portion thereof that specifically binds to.

Human CD2 as measured by surface plasmon resonance (SPR) or flow cytometry
It specifically binds to 7 and does not block the binding of its CD70 ligand, where
(a) About 100 nM or less, about 90 nM or less, about 80 nM or less, about 70 nM or less, about 60 nM or less, about 50 nM or less, about 40 nM or less, about 1 nM to about 100 nM, about 10 nM to about 70 nM, on human CD27.
Bind with a KD of about 10 nM to about 50 nM or about 40 nM to about 45 nM and / or
(b) About 0.5 nM or less, about 0.4 nM or less, about 0.3 nM or less, about 0.005 nM to about 0.5 nM, about 0.5 nM in naive human T cells stimulated with anti-CD3 antibody and anti-CD28 antibody. 01nM ~ about 0.4
Induces NF-κB and MAPK signaling with an EC50 of nM or about 0.02 nM to about 0.25 nM.
The monoclonal antibody according to claim 1 or an antigen-binding portion thereof.

( A) CDR1 containing a continuously linked amino acid having the sequence shown in SEQ ID NO: 2, CDR2 containing a continuously bound amino acid having the sequence shown in SEQ ID NO: 3, and continuously having the sequence shown in SEQ ID NO: 4. Includes heavy chain variable region (VH) containing CDR3 containing bound amino acids;
(b) CDR1 containing a continuously linked amino acid having the sequence shown in SEQ ID NO: 5, CDR2 containing a continuously bound amino acid having the sequence shown in SEQ ID NO: 6, and continuously having the sequence shown in SEQ ID NO: 7. Contains a light chain variable region (VL) containing CDR3 containing bound amino acids;
(c) Containing VHs comprising continuously linked amino acids having the sequence set forth in SEQ ID NO: 8;
(d) Includes a VL containing continuously linked amino acids having the sequence set forth in SEQ ID NO: 9;
(e) Includes a heavy chain containing continuously linked amino acids having the sequence set forth in SEQ ID NO: 12;
(f) Containing a light chain comprising a continuously linked amino acid having the sequence set forth in SEQ ID NO: 13; and / or
(g) A monoclonal antibody named BMS-986215,
The monoclonal antibody according to claim 1 or 2, or an antigen-binding portion thereof.

A monoclonal antibody or an antigen-binding portion thereof that specifically binds to the same epitope of human CD27 as the monoclonal antibody according to any one of claims 1 to 3 or an antigen-binding portion thereof.

(a) CD2 as measured by surface plasmon resonance (SPR) or flow cytometry
Does not inhibit the binding of CD70 to 7;
(b) Does not specifically bind to rat CD27 and / or mouse CD27 as measured by SPR or flow cytometry;
(c) Does not specifically bind to one or more of the TNF receptor superfamily members selected from the group consisting of CD30, HVEM, DR5, 4-1BB, CD40, OX40, GITR and any combination thereof;
(d) It does not specifically bind to one or more of the human tissues at concentrations up to 10 μg / mL, where the human tissues are thyroid, lung, skin, uterus, prostate, liver, kidney, pancreas, adrenal gland, pituitary gland, Selected from the group consisting of the placenta, testis, cerebrum, cerebellum, heart, peripheral nerves and any combination thereof;
(e) Can induce NF-κB and MAPK signaling in naive, pre-activated human T cells stimulated with anti-CD3 Ab and anti-CD28 Ab;
(f) Proliferation and / or IFN-γ in the CHO-svCD3-CD32A assay
Can induce secretion;
(g) Can induce proliferation of CD4 + CD45RO + memory T cells;
(h) Can induce increased IL-2 release from staphylococcal enterotoxin B (SEB) -stimulated human peripheral blood mononuclear cells (PBMC);
(i) Increase IL-2 release by at least 2-fold when combined with anti-planned death 1 (anti-PD-1) antibody;
(j) Invert Treg-mediated inhibition of monocyte-derived dendritic cells (MDDCs) and co-cultured CD4 + responder T cells in the presence of soluble OKT3;
(K) The monoclonal antibody according to any one of claims 1 to 4, or an antigen-binding moiety thereof, which has one or more characteristics selected from being capable of enhancing human T cell proliferation and induction of IFN-γ secretion by soluble CD70. ..

Monoclonal antibodies or antigen-binding portions thereof are:
(a) For human T cells, about 0.1 nM or less, about 0.09 nM or less, about 0.08 nM or less, about 0.07 nM or less, about 0.06 nM or less, about 0.05 nM or less, about 0.04 nM or less, about. 0.01nM ~ 0.1nM,
Bond with an EC50 of about 0.025 nM to about 0.075 nM or about 0.03 nM to about 0.06 nM;
(b) About 0.5 nM or less, about 0.4 nM or less, about 0.3 nM or less, about 0.2 nM for cynomolgus monkey T cells
Hereinafter, about 0.1 nM or less, about 0.01 nM to about 0.5 nM, about 0.025 nM to about 0.4 nM, about 0.04.
Bond with an EC50 of ~ 0.3 nM or about 0.06 to about 0.2 nM; and / or
(c) Containing heavy chain constant regions of human IgG1, IgG2, IgG3 or IgG4 isotypes.
The monoclonal antibody according to any one of claims 1 to 5 or an antigen-binding portion thereof.

(a) Human antibody or fragment thereof;
(b) Humanized antibody or fragment thereof;
(c) Chimeric antibody or fragment thereof;
(d) Human CD27 agonist;
(e) Does not inhibit the binding of CD70 to CD27 as measured by SPR or flow cytometry;
(f) Does not specifically bind to rat CD27 and / or mouse CD27 as measured by SPR or flow cytometry;
(g) Does not specifically bind to one or more of the TNF receptor superfamily members selected from the group consisting of CD30, HVEM, DR5, 4-1BB, CD40, OX40, GITR and any combination thereof;
(h) Does not specifically bind to one or more of the human tissues at concentrations up to 10 μg / mL, where the human tissues are thyroid, lung, skin, uterus, prostate, liver, kidney, pancreas, adrenal gland, pituitary gland, Selected from the group consisting of the placenta, testis, cerebrum, cerebellum, heart, peripheral nerves and any combination thereof;
(i) NF-κB and MAPK signaling in naive, pre-activated human T cells stimulated with anti-CD3 Ab and anti-CD28 Ab, optionally less than about 0.5 nM, about 0.4.
Induce with an EC50 of nM or less, about 0.3 nM or less, about 0.005 nM to about 0.5 nM, about 0.01 nM to about 0.4 nM or about 0.02 to about 0.25 nM;
(j) Proliferation and / or IFN-γ in the CHO-svCD3-CD32A assay
Secretion is preferably about 0.05 nM or less, about 0.04 nM or less, about 0.03 nM or less, about 0.000.
5nM to about 0.05nM, about 0.0005nM to about 0.04nM, about 0.0005nM to about 0.03nM,
About 0.001nM to about 0.05nM, about 0.001nM to about 0.04nM or about 0.001nM to about 0.
Induce with an EC50 of 03 nM;
(k) CD4 + CD45RO + memory T cell proliferation is preferably about 0.01 nM or less, about 0.
009nM or less, about 0.008nM or less, about 0.007nM or less, about 0.006nM or less, about 0.00
5nM or less, about 0.001nM to about 0.01nM, about 0.002nM to about 0.008nM, about 0.003n
Induce with an EC50 of M to about 0.007 nM or about 0.004 nM to about 0.006 nM;
(l) SEB-stimulated IL-2 release from human PBMCs is stimulated at least about 50% in the presence of a crosslinker, or at least about 2-fold when optionally combined with an anti-PD-1 antibody;
(m) Tr of co-cultured CD4 + responder T cells in the presence of MDDC and soluble OKT3
Eg-mediated inhibition is optionally reversed by at least about 70%; or
(n) Induction of human T cell proliferation and IFN-γ secretion by soluble CD70 is preferably about 0.01 nM or less, about 0.009 nM or less, about 0.008 nM or less, about 0.007 nM or less, about 0.
Augmented with an EC50 of .006 nM or less, about 0.005 nM or less, about 0.001 nM to about 0.01 nM, about 0.002 nM to about 0.008 nM or about 0.003 nM to about 0.005 nM and optionally IFN-γ Induction of secretion is about 0.01 nM or less, about 0.009 nM or less, about 0.008 nM or less,
Augmented with an EC50 of about 0.007 nM or less, about 0.001 nM to about 0.01 nM, about 0.002 nM to about 0.008 nM or about 0.005 nM to about 0.007 nM.
The monoclonal antibody according to any one of claims 1 to 6 or an antigen-binding portion thereof.

(A) Is it a full-length antibody of the IgG1, IgG2, IgG3 or IgG4 isotype; or
(B) The antigen-binding moiety according to any one of claims 1 to 7, which is an antibody fragment or a single-chain antibody, wherein the antibody fragment is Fab, F (ab') 2, Fd and, if desired. Select from the group consisting of Fv fragments, single domain antibodies, single chain variable fragments (scFv), divalent scFv (di-scFv) and vivalent scFv (bi-scFv), diabody, minibody, CDR, and combinations thereof. The antigen-binding portion that is to be.

An immunoconjugate comprising the monoclonal antibody according to any one of claims 1 to 8 bound to a therapeutic agent or an antigen-binding portion thereof, wherein the therapeutic agent is an immunoconjugate which is a cytotoxin or a radioisotope, if desired. Gate.

A bispecific molecule containing the monoclonal antibody according to any one of claims 1 to 8 or an antigen-binding moiety thereof, which is bound to a binding domain having a binding specificity different from that of the monoclonal antibody or the antigen-binding moiety thereof. , Bispecific molecule.

(A) The monoclonal antibody according to any one of claims 1 to 8 or an antigen-binding portion thereof;
(B) The immunoconjugate according to claim 9; or
(C) The bispecific molecule according to claim 10.
And a composition comprising a pharmaceutically acceptable carrier.

Further comprising an additional antibody or antigen-binding portion thereof, wherein the additional antibody or antigen-binding portion thereof is an anti-PD-1 antibody, an anti-planned death ligand-1 (PD-L1) antibody, an anti-cellular toxic T lymphocyte antigen-4 (CTLA). -4) Antibodies, anti-lymphocyte activation gene-3 (LAG-3) antibody, anti-B and T lymphocyte attenuator (BTLA) antibody, anti-T cell immunoglobulin and mutin domain-3 (TIM-3) antibody , Anti-killer immunoglobulin-like receptor (KIR) antibody, anti-killer cell lectin-like receptor G1 (KLRG-1) antibody, anti-adenosine A2a receptor (A2aR)
Antibodies, anti-natural killer cell receptor 2B4 (CD244) antibodies, anti-CD160 antibodies, Ig
And T cell immune receptor (TIGIT) antibody with ITIM domain, V- for T cell activation
Domain Ig Suppressor (VISTA) antibody, nibolumab, pembrolizumab, semiprimab, atezolizumab, durvalumab, avelumab, antibody named BMS-936559, anti-inducible T cell costimulatory molecule (ICOS) antibody, anti-CD137 (4-1BB) antibody , Anti-CD134 (OX40) antibody, anti-CD27 antibody, anti-glucocorticoid-inducible TNFR-related protein (GITR) antibody, anti-herpesvirus invading mediator (HVEM) antibody and any combination thereof. Item 12. The composition according to Item 11 .

An isolated nucleic acid encoding the monoclonal antibody according to any one of claims 1 to 8 or an antigen-binding portion thereof.

An expression vector containing the isolated nucleic acid according to claim 13.

A host cell comprising the expression vector according to claim 14.

A transgenic mouse containing a human immunoglobulin heavy chain and a light chain transgene, which expresses the monoclonal antibody according to any one of claims 1 to 8.

A hybridoma obtained from the mouse according to claim 16, which produces a monoclonal antibody.

A method for producing an anti-CD27 monoclonal antibody or an antigen-binding portion thereof, wherein the monoclonal antibody or the antigen-binding portion thereof is expressed in the host cell according to claim 15, and the monoclonal antibody or the antigen-binding portion thereof is expressed. A method comprising isolating from a host cell.

The monoclonal antibody or antigen-binding portion thereof according to any one of claims 1 to 8, which is a pharmaceutical composition for treating a subject having cancer, and the immunoconjugate according to claim 9. A pharmaceutical composition comprising the bispecific molecule of claim 10 or the composition of claim 11 or 12 , where the subject is optionally human .

A pharmaceutical composition for inhibiting tumor cell growth in a subject, the monoclonal antibody according to any one of claims 1 to 8 or an antigen-binding portion thereof , and the immunoconjugate according to claim 9. , A pharmaceutical composition comprising the bispecific molecule of claim 10 or the composition of claim 11 or 12 , where the subject is optionally human .

The pharmaceutical composition according to claim 19 or 20, further comprising a therapeutically effective amount of an additional therapeutic agent for the treatment of a subject with cancer or for inhibiting the growth of tumor cells in the subject , optionally. A pharmaceutical composition in which the further therapeutic agent is (a), (b), (c) or (d) below:
(A) Compounds that reduce or increase irritation of the immune system ;
(B) Small molecule compounds, macrocyclic peptides, fusion proteins, or antibodies;
(C) Planned death-1 (PD-1), Planned death ligand-1 (PD-L1), cytotoxic T lymphocyte antigen-4 (CTLA-4), lymphocyte activation gene-3 (LAG-3) , B and T lymphocyte attenuator (BTLA), T cell immunoglobulin and mutin domain-3 (TIM-3), killer immunoglobulin-like receptor (KIR), killer cell lectin-like receptor G1 (KLRG-1) , Adenosine A2a Receptor (A2aR), Natural Killer Cell Receptor 2B4 (CD244), CD160, T Cell Immunoreceptor with Ig and ITIM Domains (TIGIT), or V Domain Ig Suppressor (VISTA) for T Cell Activation An antagonist antibody or antigen-binding portion thereof that specifically binds to a receptor, wherein the antibody or antigen-binding portion thereof is one that specifically binds to PD-1 or PD-L1 and the antibody or antigen thereof. The joint is:
(I) Does it disrupt the interaction between PD-1 and PD-L1 and inhibit PD-1 / PD-L1 signaling;
(Ii) Is it a chimeric, humanized or human monoclonal antibody or antigen-binding portion thereof;
(Iii) Cross-competition with nivolumab for binding to human PD-1;
(Iv) Whether it specifically binds to PD-1 and is nivolumab or pembrolizumab; or
(V) It specifically binds to PD-L1 and is atezolizumab, durvalumab or avelumab;
(D) Specific to inducible T cell costimulatory molecule (ICOS), CD137 (4-1BB), CD134 (OX40), CD27, glucocorticoid-inducible TNFR-related protein (GITR), or herpesvirus invasion mediator (HVEM). Agonist antibody that binds to or its antigen binding site.

(A) The pharmaceutical composition according to any one of claims 19 to 21 , wherein the cancer is a solid tumor or the tumor cells are cells of a solid tumor, and the solid tumor is optionally colon cancer, fibrosarcoma, flattened. Epithelial cell cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), squamous epithelial NSCLC, non-flat epithelial NSCLC, head and neck cancer, breast cancer, esophageal cancer, gastric cancer, gastrointestinal cancer, small intestinal cancer, liver cancer, liver Cellular cancer (HCC), pancreatic cancer (PAC), kidney cancer, renal cell cancer (RCC), bladder cancer, urinary tract cancer, urinary tract cancer, colon-rectal cancer (CRC), colon cancer, anal cancer, endometrial cancer, Prostate cancer, neuroblastoma, glioma, glioma, blast cell tumor, pediatric sarcoma, sinus natural killer, melanoma, skin cancer, bone cancer, cervical cancer, uterine cancer, endometrial cancer, egg Tube cancer, ovarian cancer, cervical cancer, vaginal cancer, genital cancer, testis cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, penis cancer, renal pelvis cancer, central nervous system (CNS) neoplasm , Primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain cancer, brain stem glioma, pituitary adenoma, capsicum sarcoma, epidermal cancer, squamous cell carcinoma, pediatric solid tumor, environment-induced cancer, virus-related cancer, virus Is it a cancer of origin, advanced cancer, unresectable cancer, metastatic cancer, refractory cancer, recurrent cancer or any combination of these ; or
(B) The cancer is a hematopoietic tumor or the tumor cells are hematopoietic tumor cells .
If desired, hematopoietic tumors include T-cell lymphoma, acute lymphoblastic leukemia (ALL), acute myeloma leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloma leukemia (CML), T-cell lymphoma, Hodgkin. Lymphoma (HL), non-Hodgkin's lymphoma (NHL), multiple myeloma, smoldering myeloma, unclear monoclonal gamma globulinemia (MGUS), progressive, metastatic, refractory and / or recurrent blood A pharmaceutical composition selected from any combination of systemic tumors and the blood system tumors.

A kit for treating a subject with cancer or for inhibiting the growth of tumor cells in the subject .
(a) The monoclonal antibody according to any one of claims 1 to 8 or an antigen-binding portion thereof in the range of about 0.1 to about 20 mg / kg body weight, or the pharmaceutical composition according to claim 11 or 12 .
(B) One or more doses of additional therapeutic agent ; and
(C) A kit comprising instructions for using the one or more dose in the pharmaceutical composition according to any of claims 19-22.