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BR-112018002877-B1 - HUMANIZED ANTI-CCR7 RECEPTOR ANTIBODIES

BR112018002877B1BR 112018002877 B1BR112018002877 B1BR 112018002877B1BR-112018002877-B1

Abstract

HUMANIZED ANTI-CCR7 RECEPTOR ANTIBODIES. This disclosure provides humanized anti-CCR7 human antibodies and compositions comprising such antibodies. The antibodies and compositions are useful in the treatment of cancer in which the tumor cells express a CCR7 receptor, in the treatment of inflammatory conditions, conditions or complications arising from tissue or organ transplantation, and conditions or complications resulting from or associated with fibrosis. Experimental results are detailed for some antibodies derived from mouse antibodies. The mouse antibody with the internal name mab729 [having CDRs VH1: GLTFRDFA; VH2: ISSGGFYT, VH3: VRRAYRYDGTGDYSALDY, VL1: QDIGPS, VL2: ATS; VL3: LQFASSPLT] is also humanized (internal name mAB 650_H 1L1). This antibody inhibits the internalization of the CCR7 receptor, as shown by experimental results.

Inventors

  • JAAP WILLEM BACK
  • Ronald Boshuizen
  • WOUTER CORNELIS PUIJK
  • Johan Turkstra

Assignees

  • PEPMAB B.V

Dates

Publication Date
20260310
Application Date
20160810
Priority Date
20150810

Claims (13)

  1. 1. Humanized anti-CCR7 antibody characterized in that it comprises the hypervariable regions HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3, wherein: HVR-H1 consists of SEQ ID NO: 6; HVR-H2 consists of SEQ ID NO: 13; HVR-H3 consists of SEQ ID NO: 19; HVR-L1 consists of SEQ ID NO: 27; HVR-L2 consists of SEQ ID NO: 31; eHVR-L3 consists of SEQ ID NO: 36, and the antibody possesses at least one of the following: a) a minimum affinity for a synthetic antigen with the amino acid sequence of SEQ ID NO: 76, defined by a Kd that is no more than a factor of 10 greater than the Kd of a mouse anti-CCR7 antibody, of which the amino acid sequence of the variable heavy chain domain is SEQ ID NO: 1 and of which the amino acid sequence of the variable light chain domain is SEQ ID NO: 2; and b) an IC50 value of no more than 100 nM to inhibit at least one of CCR7-dependent intracellular signaling and CCR7 receptor internalization, in at least one CCR7 ligand selected from CCL19 and CCL21.
  2. 2. Humanized anti-CCR7 antibody, according to claim 1, characterized in that the variable heavy chain domain of the antibody comprises 4 heavy chain structural regions, HFR1 to HFR4, and 3 hypervariable regions HVR-H1 to HVR-H3 that are operatively linked in the order HFR1, HVR-H1, HFR2, HVR-H2, HFR3, HVR-H3 and HFR4, wherein the variable light chain domain of the antibody comprises 4 light chain structural regions, LFR1 to LFR4, and 3 hypervariable regions HVR-L1 to HVR-L3 that are operatively linked in the order LFR1, HVR-L1, LFR2, HVR-L2, LFR3, HVR-L3 and LFR4, wherein the heavy chain structural regions HFR1 to HFR4 have the amino acid sequences of: i) SEQ ID NOs: 40, 43, 45 and 48, respectively; ii) SEQ ID NOs: 41, 44, 46 and 49, respectively; or, iii) SEQ ID NOs: 42, 44, 47 and 49, respectively, and wherein the structural light chain regions LFR1 to LFR4 have the amino acid sequences of: iv) SEQ ID NOs: 50, 52, 55 and 58, respectively; or, v) SEQ ID NOs: 51, 53, 56 and 59, respectively.
  3. 3. Humanized anti-CCR7 antibody, according to claim 1 or 2, characterized in that the variable heavy chain domain of the antibody comprises an amino acid sequence consisting of at least one of the SEQ IDs NOs: 61, 62 and 63 and in that the variable light chain domain of the antibody comprises an amino acid sequence consisting of at least one of the SEQ IDs NOs: 64 and 65, wherein preferably the variable heavy chain domain of the antibody consists of the amino acid sequence of SEQ ID NO: 61 and preferably the variable light chain domain of the antibody consists of the amino acid sequence of SEQ ID NO: 64.
  4. 4. Humanized anti-CCR7 antibody, according to claim 3, characterized in that the variable heavy chain domain of the antibody consists of the amino acid sequence SEQ ID NO: 61 and the variable light chain domain of the antibody consists of the amino acid sequence SEQ ID NO: 64.
  5. 5. Humanized anti-CCR7 antibody, according to any one of claims 1 to 4, characterized in that the antibody comprises a constant heavy chain region that is the IgG1, IgG2, IgG3 or IgG4 region.
  6. 6. Humanized anti-CCR7 antibody, according to any one of claims 1 to 5, characterized in that the antibody comprises a functional Fc region having at least one effector function selected from the group consisting of: C1q binding, complement-dependent cytotoxicity; Fc receptor binding, antibody-dependent cell-mediated cytotoxicity and phagocytosis.
  7. 7. Humanized anti-CCR7 antibody, according to any one of claims 1 to 6, characterized in that the antibody comprises a heavy chain constant region of the G1m17,1 allotype, wherein, preferably, the heavy chain constant region comprises an E333A substitution.
  8. 8. Pharmaceutical composition characterized in that it comprises a humanized anti-CCR7 antibody as defined in any one of claims 1 to 7.
  9. 9. Use of a humanized anti-CCR7 antibody as defined in any one of claims 1 to 7, characterized in that it is for the manufacture of a medicament for the treatment of a cancer, an inflammatory condition, a condition or complication resulting from tissue or organ transplantation, or a condition or complication resulting from, or associated with, fibrosis.
  10. 10. Use of a pharmaceutical composition as defined in claim 8, characterized in that it is in the manufacture of a medicament for the treatment of cancer, an inflammatory condition, a condition or complication resulting from tissue or organ transplantation, or a condition or complication resulting from, or associated with, fibrosis.
  11. 11. Use, according to claim 9 or 10, characterized in that the cancer is a cancer of which the tumor cells express a CCR7 receptor, preferably, the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), follicular lymphoma, large B-cell lymphoma, AIDS-associated lymphoma, lymphoplasmacytic lymphoma, Burkitt's lymphoma, acute lymphoblastic leukemia of B cells, Hodgkin's disease, adult T-cell leukemia/lymphoma, mycosis fungoides, blast crisis of chronic myeloproliferative syndromes, blast crisis of myelodysplastic syndromes, breast cancer, non-small cell lung cancer, melanoma, gastric cancer, squamous cell carcinoma of the head and neck, and colon carcinoma.
  12. 12. Use, according to claim 9 or 10, characterized in that the inflammatory condition is selected from the group consisting of inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, allergic inflammation of the airways, airway smooth muscle hyperplasia, fibrotic lung disease, rheumatoid arthritis, multiple sclerosis, psoriasis, atherosclerosis, HIV infection and AIDS, or in which the tissue or organ transplant is one or more kidney, heart, skin and lung transplants.
  13. 13. Use, according to claim 9 or 10, characterized in that the fibrosis is selected from the group consisting of hepatic fibrosis and hepatic cirrhosis, renal fibrosis, pulmonary fibrosis, skin fibrosis, cardiovascular fibrosis, gastrointestinal fibrosis.

Description

Field of invention [001] The present invention relates generally to the fields of medicine and pharmacy, in particular to the field of biopharmaceuticals for use in oncology. More specifically, the invention relates to CCR7 receptor antibodies that are useful in the treatment of cancers in which tumor cells express a CCR7 receptor. Fundamentals of the invention [002] The human CC7 motif receptor (hereinafter referred to as “CCR7”) is a 7-transmembrane spanning domain G protein-coupled receptor (GPCR) that was originally found to be expressed in a lymphocyte-selective manner by EBV infection (Birkenbach et al., 1993, J. Virol. 67: 2209-2220). Later, CCR7 was shown to be a selective chemokine receptor for CCL19 and CCL21. Under physiological conditions, CCR7 expression is restricted to naive T and B lymphocytes and mature dendritic cells and plays a role in regulating their migration, organization, and activation. [003] CCR7 activity has been implicated in a wide variety of disease states, including chronic inflammatory conditions (Moschovakis et al, 2012, Eur J Immunol. 42: 1949-1955), atherosclerosis (Luchtefeld et al, 2010, Circulation 122: 1621-1628), HIV infection (Evans et al, 2012, Cytokine Growth Factor Rev. 23: 151-57) and cancer (Ben-Baruch, 2009, Cell Adhesion Migration 3: 32833). [004] Several studies have revealed that CCR7 is expressed in a wide variety of tumor cells, including, for example, mantle cell lymphoma (MCL), follicular lymphoma, large B-cell lymphoma, AIDS-associated lymphoma, lymphoplasmacytic lymphoma, Burkitt lymphoma, acute lymphoblastic leukemia of B cells, Hodgkin's disease, adult T-cell leukemia/lymphoma, mycosis fungoides, blast crisis of chronic myeloproliferative syndromes, blast crisis of myelodysplastic syndromes, cancers such as breast cancer, non-small cell lung cancer, melanoma, gastric cancer or squamous cell carcinoma of the head and neck and colon carcinoma such as chronic lymphocytic leukemia of B cells, non-Hodgkin's lymphoma, breast cancer cells and malignant breast tumor (WO 2007/003426). Furthermore, it becomes clear that CCR7 plays a role in lymph node metastasis of various cancers (Viola and Luster, 2008, Annu Rev Pharmacol Toxicol. 48: 171-97). [005] A reference mouse monoclonal antibody against human CCR7 (MAB197, clone # 150503) is commercially available from R&D Systems (www.RnDSystems.com). [006] Document WO 2007/003426 discloses the use of an antibody that binds to CCR7 to treat cancer in which tumor cells express CCR7. The binding of the antibody to CCR7-expressing tumor cells is disclosed to inhibit tumor cell migration and/or to kill tumor cells by one or more of complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), and inducing apoptosis. [007] Document WO 2012/043533 discloses 8 different mouse monoclonal antibodies against human CCR7 that are useful for the treatment of fibrosis or cancer. However, none of these antibodies has an IC50 of less than 7.4 nM for the inhibition of intracellular Ca2+ signal transduction. [008] Document WO 2014/151834 discloses three different human anti-CCR7 antibodies generated in a Xenomous™ immunized with human cells expressing CCR7, as well as a mouse human anti-CCR7 antibody and chimerization and humanization thereof. The human anti-CCR7 antibodies bind to a different epitope on human CCR7 than the reference mouse monoclonal antibody MAB197. The antibodies are disclosed as being useful for the treatment of inflammatory conditions, cancerous conditions, as well as conditions and complications arising from tissue or organ transplantation. Document WO 2014/151834 further discloses the isolation of mouse human anti-CCR7 antibodies, including the MAb 22 blocking antibody, which was chimerized by replacing its heavy chain constant regions with its human homologs. This chimerized antibody retained its ability to bind to human CCR7. The light chain of this chimerized antibody was further altered to generate a series of MAb 22 variants with chimerized heavy chains and humanized light chains. WO 2014/151834 does not disclose whether some of these MAb 22 variants with humanized light chains retained the ability to bind to human CCR7. [009] There is, however, still a need in the art for other and better anti-CCR7 antibodies that are useful in human therapy. Summary of the invention [010] In a first aspect, the invention relates to a humanized anti-CCR7 antibody comprising the hypervariable regions HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3, wherein: HVR-H1 comprises the sequence: G-F/L-T/A/P-F-S/T/R-N/D/S-Y/F-A; HVR-H2 comprises the sequence: I-S-S/D-G/R-G-S/T/F-Y/H/F-T/P; HVR-H3 comprises the sequence: A/T/V/G-R-R/A-A/E/T-Y/G/T-R/V-Y/V-D/*-GTG/*-E/V/D/A/G/*-N/S/D/T-A/S/D-M/L/F-Y/S; HVR-L1 comprises the sequence: Q/S-D/S-I/L/V-G/S/L-D/S/P/G/N-S/N-*/Y/DGKTY; HVR-L2 comprises the sequence: A/S/T-T/I/V-S; and HVR-L3 comprises the sequence: L/W/Q-Q-Y/F/G/W-A/T/S-S/N/H-S/F/N-P-L/P/Q-T