A method of treating cancer using anti-MICA / B antibodies.

Abstract

This specification provides a method for treating cancer using an antibody or antigen-binding fragment thereof that specifically binds to MICA / B, having a heavy chain, a light chain, a heavy chain variable domain (VH), a light chain variable domain (VL), and a complementarity-determining region (CDR) as disclosed herein. In a second aspect, this disclosure provides a method for treating parotid gland cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (an anti-MICA / B antibody).

Description

[Technical Field]

[0001] Related applications This patent application claims priority to U.S. Provisional Patent Application No. 63 / 504,163 filed on 24 May 2023, U.S. Provisional Patent Application No. 63 / 505,705 filed on 1 June 2023, U.S. Provisional Patent Application No. 63 / 593,609 filed on 27 October 2023, U.S. Provisional Patent Application No. 63 / 559,422 filed on 29 February 2024, and U.S. Provisional Patent Application No. 63 / 574,800 filed on 4 April 2024, the contents of which are thus incorporated in their entirety by reference into this disclosure.

[0002] Reference to electronic sequence listings The contents of the electronic sequence listing (67239WO01_SequenceListing.xml, size: 14,820 bytes, and creation date: May 13, 2024) are incorporated herein by reference in their entirety. [Background technology]

[0003] The field of immuno-oncology has brought about a major transformation in the treatment of cancer patients, improving patient survival and quality of life. However, to date, much of this field has focused on leveraging the power of the adaptive immune response through the therapeutic targeting of T cells. While these approaches have made significant progress in the field, several challenges remain. Furthermore, the clinical utility of T-cell therapy does not extend to all patients or tumor indications, particularly in the context of solid tumors. Alternative strategies, such as involving the innate immune system, have become a focus of considerable attention in this field. In particular, the involvement of natural killer (NK) cells as potent effectors of the innate immune response has emerged as a promising modality in immunotherapy. Whalen et al., 2023, provides a detailed review of the potential of involving NK cells in cancer therapies using NKG2D, CD16A, and other cell surface receptors (Whalen et al., mAbs 15:12208697).

[0004] In contrast to T cells, natural killer (NK) cells act as the front line of defense, possessing the ability to detect and eliminate tumor or virus-infected cells without any prior antigen-specific activation or differentiation. Therefore, NK cells possess several compelling properties that justify the development of drugs that leverage their therapeutic potential. The complex balance between activating and inhibitory signals characteristic of NK cells necessitates the involvement of multiple activating receptors, the combination of receptor agonists and NK cell checkpoint inhibitors, and / or combination with T cell therapy to maximize therapeutic efficacy in patients. Similar to T cells, NK cells are increasingly recognized as possessing high cytotoxicity when properly bound. NK cell therapy has been shown to promote complete response in patients when equipped with potent activating receptors specific to cell surface antigens. Similarly, T cell receptor complexes (TCRs) in NK cells enhance their effector function in a manner similar to that of T cells.

[0005] NK cell therapy offers several advantages over T cell therapy. Most notably, their allogeneic and immediate use potential is significant, as they can avoid graft-versus-host reactivity due to the absence of TCRs. In addition, NK cell therapy is typically associated with less severe toxicity, such as immune effector cell-associated neurotoxicity (ICANs) and cytokine release syndrome (CRS), compared to certain T cell-based therapies. However, NK cell-based cell therapies still face several significant challenges, including inefficient transgene delivery, the complexity of NK cell production, and the exhaustion and limited persistence of genetically engineered NK cells.

[0006] In fact, cancer therapy has been dramatically transformed by the development of antibodies that specifically target T cells rather than cancer cells, in order to effectively treat cancer patients and, in some cases, provide them with a sustained response. Unlike cell therapies, mAbs that bind to NK cells reach all effector NK cells, much like those that target T cells, rely on standardized mAb manufacturing methods, allow for predictable pharmacokinetic properties, and thereby have the potential to achieve a potentially potent and pan-oncological response. This field is rich in mAb-based therapies that target the interaction between CD16A / Fc-gamma receptor IIIA (FcγRIIIa) expressed on NK cells and the Fc-gamma domain 1 (Fcγ1) of mAbs that recognize tumor-associated cell surface antigens on cancer cells. Many chimeric, human, and humanized IgG1 antibody therapies, such as rituximab (targeting CD20), daratumumab (CD38), and trastuzumab (human epidermal growth factor receptor 2-HER2), bind to CD16A. However, these conventional antibody-based therapies fail to utilize another important NK cell activation receptor, namely NKG2D. NKG2D is a potent regulator of NK cell activation that has recently emerged as a key target in the field of immuno-oncology. NKG2D recognizes eight distinct ligands that are upregulated in tumor cells: MHC class I chain-related protein A (MICA), MHC class I chain-related protein sequence B (MICB), and UL-16 binding proteins (ULBPs) designated as ULBP1-ULBP6.

[0007] While strategies for therapeutically utilizing NKG2D activity are still in the early stages of development, MICA is the most consistently and highly expressed ligand across all solid tumors and hematological malignancies compared to other NKG2D ligands. The second most widespread expression pattern is shown by MICB, which is closely related to MICA in terms of sequence. Therefore, both MICA and MICB (MICA / B) are attractive pan-oncological targets.

[0008] MICA and MICB are stress-induced surface glycoproteins that are upregulated in a wide variety of human tumors. Binding of MICA / B by the activating receptor NKG2D in several subsets of NK and T cells leads to immune-mediated target cell lysis.

[0009] However, cancer cells evade NKG2D-mediated NK cell lysis by exploiting the intrinsic alpha-3 region of MICA / B, which is the target of proteolytic shedding of the extracellular domain (ECD) from the cancer cell surface via the action of multiple proteases expressed on the cell surface or released into the tumor microenvironment by most cancer cells. There is a need for therapies that prevent MICA / B shedding and thereby conserve MICA / B on the tumor cell surface to promote NKG2D-mediated NK cell lysis against tumor cells. Such therapies may have the potential to treat a wide variety of cancers. Therefore, there is an unmet need to develop novel therapies that can treat cancers that evade the immune response using this mechanism. [Overview of the project]

[0010] A method for treating cancer using an antibody that specifically binds to MICA / B is disclosed herein.

[0011] Accordingly, in the first aspect, the present disclosure is characterized by a method for treating endometrial cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (an anti-MICA / B antibody).

[0012] In some embodiments of the first aspect, the subject has previously been treated with a PD-1 inhibitor. In some embodiments, the subject has previously been treated with a VEGF inhibitor or hormone therapy. In some embodiments, the subject has never been treated with a PD-1 inhibitor. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0013] In some embodiments of the first model, the endometrial cancer is HER2+. In some embodiments, the endometrial cancer is ER+ or PR+, or both. In some embodiments, the cancer is one or more variants of EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0014] In some embodiments of the first aspect, the subject shows a response after administration of the anti-MICA / B antibody. In some embodiments, the response is a complete response. In some embodiments, the response is a partial response. In some embodiments, the subject shows disease stabilization after administration of the anti-MICA / B antibody. In some embodiments, the response or disease stabilization after administration of the anti-MICA / B antibody lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0015] In some embodiments of the first aspect, the subject has previously been treated with VEGF inhibitors, EGFR inhibitors, and / or hormone therapy.

[0016] In some embodiments of the first aspect, the EGFR inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib. In some embodiments, the EGFR inhibitor comprises an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nesitumumab.

[0017] In some embodiments of the first aspect, the VEGF inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib. In some embodiments, the VEGF inhibitor comprises an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0018] In some embodiments of the first aspect, the hormone therapy includes an aromatase inhibitor (AI). In some embodiments, the AI ​​is anastrozole. In some embodiments, the hormone therapy includes a selective estrogen receptor modulator (SERM). In some embodiments, the hormone therapy includes a luteinizing hormone-releasing hormone (LHRH) agonist. In some embodiments, the hormone therapy includes an antiandrogen. In some embodiments, the hormone therapy includes a CYP17 inhibitor. In some embodiments, the hormone therapy includes a progestin. In some embodiments, the hormone therapy includes an anti-adrenergic agent. In some embodiments, the hormone therapy includes an estrogen receptor antagonist.

[0019] In some embodiments of the first embodiment, the subjects have cancer characterized by low levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by high levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subjects have cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subjects have immunologically cold cancer. In some embodiments, the subjects have immunologically hot cancer. In some embodiments, the subjects have hormone-sensitive cancer. In some embodiments, the subjects have cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subjects have cancer expressing one or more of EGFR, ER, PR, or HER2.

[0020] In some embodiments of the first aspect, the subject has metastatic cancer.

[0021] In some embodiments of the first aspect, the subject has previously received therapy including chemotherapy.

[0022] In some embodiments of the first aspect, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to an administration interval. In some embodiments, the administration interval is referred to as a cycle. In some embodiments, the administration interval includes a 3-week cycle, in which the anti-MICA / B antibody is administered once every 3 weeks (Q3W). In some embodiments, the administration interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times. In some embodiments, the repetition of the dosing interval is performed over a period of at least 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 3 years, 4 years, or 5 years.

[0023] In some embodiments of the first aspect, the anti-MICA / B antibody comprises a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 1, a light chain complementarity determining region (LCDR2) of SEQ ID NO: 2, and a light chain complementarity determining region 3 (LCDR3) of SEQ ID NO: 3, and a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 4, a heavy chain complementarity determining region 2 (HCDR2) of SEQ ID NO: 5, and a heavy chain complementarity determining region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, VL comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, VH comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the light chain comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the heavy chain comprises the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

[0024] In some embodiments of the first aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the anti-MICA / B antibody binds to the alpha-3 domain of the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the MICA protein is a membrane-bound MICA protein, a soluble MICA protein, or both. In some embodiments, the MICB protein is a membrane-bound MICB protein, a soluble MICB protein, or both.

[0025] In some embodiments of the first aspect, the anti-MICA / B antibody is selected from whole immunoglobulins, scFv, Fab, F(ab’)2, or disulfide-bonded Fv. In some embodiments, the anti-MICA / B antibody is IgG or IgM. In some embodiments, the anti-MICA / B antibody is a humanized or chimeric antibody.

[0026] In some embodiments of the first aspect, the method further comprises administering to the subject an effective amount of a PD-1 inhibitor. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pembrolizumab, nivolumab, cemiplimab, AMP-224, APM-514, or spartalizumab.

[0027] In a second aspect, the present disclosure features a method of treating adenocarcinoma of the parotid gland in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (anti-MICA / B antibody).

[0028] In some embodiments of the second aspect, the adenocarcinoma of the parotid gland overexpresses one or both of EGFR and HER2.

[0029] In some embodiments of the second aspect, the adenocarcinoma of the parotid gland is mucoepidermoid carcinoma of the parotid gland.

[0030] In some embodiments of the second aspect, the cancer is one or more variants of EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0031] In some embodiments of the second aspect, the subjects show a response after administration of the anti-MICA / B antibody. In some embodiments, the response is a complete response. In some embodiments, the response is a partial response. In some embodiments, the subjects show disease stabilization after administration of the anti-MICA / B antibody. In some embodiments, the response or disease stabilization after administration of the anti-MICA / B antibody lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0032] In some embodiments of the second aspect, the subject has previously been treated with VEGF inhibitors, EGFR inhibitors, and / or hormone therapy.

[0033] In some embodiments of the second aspect, the EGFR inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib. In some embodiments, the EGFR inhibitor comprises an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nesitumumab.

[0034] In some embodiments of the second aspect, the VEGF inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib. In some embodiments, the VEGF inhibitor comprises an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0035] In some embodiments of the second embodiment, the hormone therapy includes an aromatase inhibitor (AI). In some embodiments, the AI ​​is anastrozole. In some embodiments, the hormone therapy includes a selective estrogen receptor modulator (SERM). In some embodiments, the hormone therapy includes a luteinizing hormone-releasing hormone (LHRH) agonist. In some embodiments, the hormone therapy includes an antiandrogen. In some embodiments, the hormone therapy includes a CYP17 inhibitor. In some embodiments, the hormone therapy includes a progestin. In some embodiments, the hormone therapy includes an anti-adrenergic drug. In some embodiments, the hormone therapy includes an estrogen receptor antagonist.

[0036] In some embodiments of the second embodiment, the subjects have cancer characterized by low levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by high levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subjects have cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subjects have immunologically cold cancer. In some embodiments, the subjects have immunologically hot cancer. In some embodiments, the subjects have hormone-sensitive cancer. In some embodiments, the subjects have cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subjects have cancer expressing one or more of EGFR, ER, PR, or HER2.

[0037] In some embodiments of the second aspect, the subject has metastatic cancer.

[0038] In some embodiments of the second aspect, the subject has previously received therapy including chemotherapy.

[0039] In some embodiments of the second aspect, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to an administration interval. In some embodiments, the administration interval is referred to as a cycle. In some embodiments, the administration interval includes a 3-week cycle, in which the anti-MICA / B antibody is administered once every 3 weeks (Q3W). In some embodiments, the administration interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times. In some embodiments, the repetition of the dosing interval is performed over a period of at least 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 3 years, 4 years, or 5 years.

[0040] In some embodiments of the second aspect, the anti-MICA / B antibody includes a light chain variable region (VL) comprising the light chain complementarity determination region 1 (LCDR1) of SEQ ID NO: 1, the light chain complementarity determination region (LCDR2) of SEQ ID NO: 2, and the light chain complementarity determination region 3 (LCDR3) of SEQ ID NO: 3, and a heavy chain variable region (VH) comprising the heavy chain complementarity determination region 1 (HCDR1) of SEQ ID NO: 4, the heavy chain complementarity determination region 2 (HCDR2) of SEQ ID NO: 5, and the heavy chain complementarity determination region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, VL comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, VH comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the light chain includes the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the heavy chain includes the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

[0041] In some embodiments of the second aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the anti-MICA / B antibody binds to the alpha-3 domain of the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the MICA protein is a membrane-bound MICA protein, a soluble MICA protein, or both. In some embodiments, the MICB protein is a membrane-bound MICB protein, a soluble MICB protein, or both.

[0042] In some embodiments of the second aspect, the anti-MICA / B antibody is selected from whole immunoglobulin, scFv, Fab, F(ab')2, or disulfide-linked Fv. In some embodiments, the anti-MICA / B antibody is IgG or IgM. In some embodiments, the anti-MICA / B antibody is a humanized or chimeric antibody.

[0043] In some embodiments of the second aspect, the method further comprises administering an effective amount of a PD-1 inhibitor to a subject. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0044] In a third aspect, the disclosure is characterized by a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective dose of an antibody that binds to MICA / B (an anti-MICA / B antibody), wherein the subject has previously been treated with (i) a VEGF inhibitor, (ii) an EGFR inhibitor, and / or (iii) hormone therapy.

[0045] In some embodiments of the third aspect, the subjects responded to VEGF inhibitors, EGFR inhibitors, or hormone therapy. In some embodiments, the response was a complete response. In some embodiments, the response was a partial response. In some embodiments, the subjects had a response duration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0046] In some embodiments of the third aspect, the patient progressed after a period of response to a VEGF inhibitor, EGFR inhibitor, or hormone therapy. In some embodiments, administration of an anti-MICA / B antibody is performed after progression following a period of response to a VEGF inhibitor, EGFR inhibitor, or hormone therapy.

[0047] In some embodiments of the third aspect, subjects showed disease stabilization upon administration of VEGF inhibitors, EGFR inhibitors, or hormone therapy. In some embodiments, subjects had disease stabilization periods of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0048] In some embodiments of the third aspect, treatment using VEGF inhibitors, EGFR inhibitors, or hormone therapy was discontinued before the initiation of anti-MICA / B antibody administration.

[0049] In some embodiments of the third aspect, treatment using a VEGF inhibitor, an EGFR inhibitor, or hormone therapy is continued after the initiation of administration of an anti-MICA / B antibody.

[0050] In some embodiments of the third aspect, the subject's condition progressed when treated with a VEGF inhibitor, an EGFR inhibitor, or hormone therapy.

[0051] In some embodiments of the third aspect, the subjects show a response after administration of the anti-MICA / B antibody. In some embodiments, the response is a complete response. In some embodiments, the response is a partial response. In some embodiments, the subjects show disease stabilization after administration of the anti-MICA / B antibody. In some embodiments, the response or disease stabilization after administration of the anti-MICA / B antibody lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0052] In some embodiments of the third aspect, the EGFR inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib. In some embodiments, the EGFR inhibitor comprises an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nesitumumab.

[0053] In some embodiments of the third aspect, the VEGF inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib. In some embodiments, the VEGF inhibitor comprises an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0054] In some embodiments of the third embodiment, the hormone therapy includes an aromatase inhibitor (AI). In some embodiments, the AI ​​is anastrozole. In some embodiments, the hormone therapy includes a selective estrogen receptor modulator (SERM). In some embodiments, the hormone therapy includes a luteinizing hormone-releasing hormone (LHRH) agonist. In some embodiments, the hormone therapy includes an antiandrogen. In some embodiments, the hormone therapy includes a CYP17 inhibitor. In some embodiments, the hormone therapy includes a progestin. In some embodiments, the hormone therapy includes an anti-adrenergic drug. In some embodiments, the hormone therapy includes an estrogen receptor antagonist.

[0055] In some embodiments of the third aspect, cancer is a tumor of the female reproductive organs. In some embodiments, a tumor of the female reproductive organs includes an endometrial tumor. In some embodiments, a tumor of the female reproductive organs includes ovarian cancer or cervical cancer. In some embodiments, cancer is a salivary gland tumor. In some embodiments, a salivary gland tumor includes a mucoepidermoid tumor. In some embodiments, a mucoepidermoid tumor is a parotid gland tumor.

[0056] In some embodiments of the third embodiment, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is colon cancer. In some embodiments, the subject has parotid gland cancer. In some embodiments, the subject has cervical cancer. In some embodiments, the subject has endometrial cancer. In some embodiments, the subject has breast cancer. In some embodiments, the subject has colon cancer. In some embodiments, the subject has ovarian cancer. In some embodiments, the subject has prostate cancer. In some embodiments, the subject has sarcoma. In some embodiments, the subject has melanoma. In some embodiments, the subject has salivary gland adenoid cystic neoplasm. In some embodiments, the subject has peritoneal mesothelioma. In some embodiments, the subject has squamous cell carcinoma of the rectum. In some embodiments, the subject has leiomyosarcoma. In some embodiments, the subject has colorectal cancer. In some embodiments, the subject has renal cancer. In some embodiments, the subject has thyroid cancer. In some embodiments, the subject has NSCLC. In some embodiments, the subject has duodenal cancer. In some embodiments, the subject has pancreatic cancer. In some embodiments, the subject has mediastinal endometrial sarcoma. In some embodiments, the subject has head and neck cancer. In some embodiments, the subject has cecal cancer.

[0057] In some embodiments of the third embodiment, the subjects have cancer characterized by low levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by high levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subjects have cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subjects have immunologically cold cancer. In some embodiments, the subjects have immunologically hot cancer. In some embodiments, the subjects have hormone-sensitive cancer. In some embodiments, the subjects have cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subjects have cancer expressing one or more of EGFR, ER, PR, or HER2.

[0058] In some embodiments of the third aspect, the subject has metastatic cancer.

[0059] In some embodiments of the third aspect, the subject has previously received therapy including chemotherapy.

[0060] In some embodiments of the third aspect, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to an administration interval. In some embodiments, the administration interval is referred to as a cycle. In some embodiments, the administration interval includes a 3-week cycle, in which the anti-MICA / B antibody is administered once every 3 weeks (Q3W). In some embodiments, the administration interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times. In some embodiments, the repetition of the dosing interval is performed over a period of at least 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 3 years, 4 years, or 5 years.

[0061] In some embodiments of the third aspect, the anti-MICA / B antibody includes a light chain variable region (VL) comprising the light chain complementarity determination region 1 (LCDR1) of SEQ ID NO: 1, the light chain complementarity determination region (LCDR2) of SEQ ID NO: 2, and the light chain complementarity determination region 3 (LCDR3) of SEQ ID NO: 3, as well as a heavy chain variable region (VH) comprising the heavy chain complementarity determination region 1 (HCDR1) of SEQ ID NO: 4, the heavy chain complementarity determination region 2 (HCDR2) of SEQ ID NO: 5, and the heavy chain complementarity determination region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, VL comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, VH comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the light chain includes the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the heavy chain includes the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

[0062] In some embodiments of the third aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the anti-MICA / B antibody binds to the alpha-3 domain of the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the MICA protein is a membrane-bound MICA protein, a soluble MICA protein, or both. In some embodiments, the MICB protein is a membrane-bound MICB protein, a soluble MICB protein, or both.

[0063] In some embodiments of the third aspect, the anti-MICA / B antibody is selected from whole immunoglobulin, scFv, Fab, F(ab')2, or disulfide-linked Fv. In some embodiments, the anti-MICA / B antibody is IgG or IgM. In some embodiments, the anti-MICA / B antibody is a humanized or chimeric antibody.

[0064] In some embodiments of the third aspect, the method further comprises administering an effective amount of a PD-1 inhibitor to a subject. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0065] In a fourth aspect, the disclosure provides a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective dose of an antibody that binds to MICA / B (an anti-MICA / B antibody), wherein the subject has previously been treated with a PD-1 inhibitor. In some embodiments, the subject responded to the PD-1 inhibitor.

[0066] In some embodiments of the fourth aspect, the response was a complete response. In some embodiments, the response was a partial response. In some embodiments, the subjects had a response duration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months or longer. In some embodiments, the subjects progressed after the response duration to the PD-1 inhibitor.

[0067] In some embodiments of the fourth aspect, subjects showed disease stabilization upon administration of a PD-1 inhibitor. In some embodiments, subjects had disease stabilization periods of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0068] In some embodiments of the fourth aspect, the treatment using a PD-1 inhibitor was discontinued before the initiation of administration of an anti-MICA / B antibody.

[0069] In some embodiments of the fourth aspect, the treatment using a PD-1 inhibitor is continued after the initiation of administration of an anti-MICA / B antibody.

[0070] In some embodiments of the fourth aspect, the subjects show a response after administration of the anti-MICA / B antibody. In some embodiments, the response is a complete response. In some embodiments, the response is a partial response. In some embodiments, the subjects show disease stabilization after administration of the anti-MICA / B antibody. In some embodiments, the response or disease stabilization after administration of the anti-MICA / B antibody lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

[0071] In some embodiments of the fourth aspect, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0072] In some embodiments of the fourth aspect, the subject has previously been treated with VEGF inhibitors, EGFR inhibitors, and / or hormone therapy.

[0073] In some embodiments of the fourth aspect, the EGFR inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib. In some embodiments, the EGFR inhibitor comprises an anti-EGFR antibody. In some embodiments, the anti-EGFR antibody is cetuximab, panitumumab, or nesitumumab.

[0074] In some embodiments of the fourth aspect, the VEGF inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib. In some embodiments, the VEGF inhibitor comprises an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab or ranibizumab.

[0075] In some embodiments of the fourth embodiment, the hormone therapy includes an aromatase inhibitor (AI). In some embodiments, the AI ​​is anastrozole. In some embodiments, the hormone therapy includes a selective estrogen receptor modulator (SERM). In some embodiments, the hormone therapy includes a luteinizing hormone-releasing hormone (LHRH) agonist. In some embodiments, the hormone therapy includes an antiandrogen. In some embodiments, the hormone therapy includes a CYP17 inhibitor. In some embodiments, the hormone therapy includes a progestin. In some embodiments, the hormone therapy includes an anti-adrenergic drug. In some embodiments, the hormone therapy includes an estrogen receptor antagonist.

[0076] In some embodiments of the fourth aspect, cancer is a tumor of the female reproductive organs. In some embodiments, tumors of the female reproductive organs include endometrial tumors. In some embodiments, tumors of the female reproductive organs include ovarian cancer or cervical cancer. In some embodiments, cancer is a salivary gland tumor. In some embodiments, salivary gland tumors include mucoepidermoid tumors. In some embodiments, mucoepidermoid tumors are parotid gland tumors.

[0077] In some embodiments of the fourth aspect, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is colon cancer. In some embodiments, the subject has parotid gland cancer. In some embodiments, the subject has cervical cancer. In some embodiments, the subject has endometrial cancer. In some embodiments, the subject has breast cancer. In some embodiments, the subject has colon cancer. In some embodiments, the subject has ovarian cancer. In some embodiments, the subject has prostate cancer. In some embodiments, the subject has sarcoma. In some embodiments, the subject has melanoma. In some embodiments, the subject has salivary gland adenoid cystic neoplasm. In some embodiments, the subject has peritoneal mesothelioma. In some embodiments, the subject has squamous cell carcinoma of the rectum. In some embodiments, the subject has leiomyosarcoma. In some embodiments, the subject has colorectal cancer. In some embodiments, the subject has renal cancer. In some embodiments, the subject has thyroid cancer. In some embodiments, the subject has NSCLC. In some embodiments, the subject has duodenal cancer. In some embodiments, the subject has pancreatic cancer. In some embodiments, the subject has mediastinal endometrial sarcoma. In some embodiments, the subject has head and neck cancer. In some embodiments, the subject has cecal cancer.

[0078] In some embodiments of the fourth embodiment, the subjects have cancer characterized by low levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by high levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subjects have cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subjects have immunologically cold cancer. In some embodiments, the subjects have immunologically hot cancer. In some embodiments, the subjects have hormone-sensitive cancer. In some embodiments, the subjects have cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subjects have cancer expressing one or more of EGFR, ER, PR, or HER2.

[0079] In some embodiments of the fourth aspect, the subject has metastatic cancer.

[0080] In some embodiments of the fourth aspect, the subject has previously received therapy including chemotherapy.

[0081] In some embodiments of the fourth aspect, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to an administration interval. In some embodiments, the administration interval is referred to as a cycle. In some embodiments, the administration interval includes a 3-week cycle, in which the anti-MICA / B antibody is administered once every 3 weeks (Q3W). In some embodiments, the administration interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times. In some embodiments, the repetition of the dosing interval is performed over a period of at least 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 3 years, 4 years, or 5 years.

[0082] In some embodiments of the fourth aspect, the anti-MICA / B antibody includes a light chain variable region (VL) comprising the light chain complementarity determination region 1 (LCDR1) of SEQ ID NO: 1, the light chain complementarity determination region (LCDR2) of SEQ ID NO: 2, and the light chain complementarity determination region 3 (LCDR3) of SEQ ID NO: 3, and a heavy chain variable region (VH) comprising the heavy chain complementarity determination region 1 (HCDR1) of SEQ ID NO: 4, the heavy chain complementarity determination region 2 (HCDR2) of SEQ ID NO: 5, and the heavy chain complementarity determination region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, VL comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, VH comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the light chain includes the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the heavy chain includes the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

[0083] In some embodiments of the fourth aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the anti-MICA / B antibody binds to the alpha-3 domain of the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the MICA protein is a membrane-bound MICA protein, a soluble MICA protein, or both. In some embodiments, the MICB protein is a membrane-bound MICB protein, a soluble MICB protein, or both.

[0084] In some embodiments of the fourth aspect, the anti-MICA / B antibody is selected from whole immunoglobulin, scFv, Fab, F(ab')2, or disulfide-linked Fv. In some embodiments, the anti-MICA / B antibody is IgG or IgM. In some embodiments, the anti-MICA / B antibody is a humanized or chimeric antibody.

[0085] In some embodiments of the fourth aspect, the method further comprises administering an effective amount of a PD-1 inhibitor to a subject. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0086] In a fifth aspect, the disclosure is characterized by a method for treating cancer in a subject, comprising (i) administering an initial dose of an antibody that binds to MICA / B (an anti-MICA / B antibody) to the subject, and (ii) administering a therapeutically effective dose of a corticosteroid to the subject prior to the initial dose of the anti-MICA / B antibody.

[0087] In some embodiments of the fifth aspect, the corticosteroid is administered 30 to 60 minutes before the administration of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered in a dose of about 2 mg to about 50 mg. In some embodiments, the corticosteroid is administered in a dose of about 10 mg. In some embodiments, the corticosteroid is administered orally or intravenously.

[0088] In some embodiments of the fifth aspect, the method further includes administering one or more subsequent doses of an anti-MICA / B antibody.

[0089] In some embodiments of the fifth aspect, a corticosteroid is administered before the initial dose of the anti-MICA / B antibody, but not before one or more subsequent doses of the anti-MICA / B antibody.

[0090] In some embodiments of the fifth aspect, the corticosteroid is dexamethasone.

[0091] In some embodiments of the fifth aspect, cancer is a tumor of the female reproductive organs. In some embodiments, a tumor of the female reproductive organs includes an endometrial tumor. In some embodiments, a tumor of the female reproductive organs includes ovarian cancer or cervical cancer. In some embodiments, cancer is a salivary gland tumor. In some embodiments, a salivary gland tumor includes a mucoepidermoid tumor. In some embodiments, a mucoepidermoid tumor is a parotid gland tumor.

[0092] In some embodiments of the fifth aspect, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is colon cancer. In some embodiments, the subject has parotid gland cancer. In some embodiments, the subject has cervical cancer. In some embodiments, the subject has endometrial cancer. In some embodiments, the subject has breast cancer. In some embodiments, the subject has colon cancer. In some embodiments, the subject has ovarian cancer. In some embodiments, the subject has prostate cancer. In some embodiments, the subject has sarcoma. In some embodiments, the subject has melanoma. In some embodiments, the subject has salivary gland adenoid cystic neoplasm. In some embodiments, the subject has peritoneal mesothelioma. In some embodiments, the subject has squamous cell carcinoma of the rectum. In some embodiments, the subject has leiomyosarcoma. In some embodiments, the subject has colorectal cancer. In some embodiments, the subject has renal cancer. In some embodiments, the subject has thyroid cancer. In some embodiments, the subject has NSCLC. In some embodiments, the subject has duodenal cancer. In some embodiments, the subject has pancreatic cancer. In some embodiments, the subject has mediastinal endometrial sarcoma. In some embodiments, the subject has head and neck cancer. In some embodiments, the subject has cecal cancer.

[0093] In some embodiments of the fifth embodiment, the subjects have cancer characterized by low levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by high levels of PD-L1 expression. In some embodiments, the subjects have cancer characterized by low tumor mutational burden (TMB). In some embodiments, the subjects have cancer characterized by high tumor mutational burden (TMB). In some embodiments, the subjects have immunologically cold cancer. In some embodiments, the subjects have immunologically hot cancer. In some embodiments, the subjects have hormone-sensitive cancer. In some embodiments, the subjects have cancer characterized by overexpression of oncogenic drivers. In some embodiments, the subjects have cancer expressing one or more of EGFR, ER, PR, or HER2.

[0094] In some embodiments of the fifth aspect, the subject has metastatic cancer.

[0095] In some embodiments of the fifth aspect, the subject has previously received therapy including chemotherapy.

[0096] In some embodiments of the fifth aspect, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered to the subject according to an administration interval. In some embodiments, the administration interval is referred to as a cycle. In some embodiments, the administration interval includes a 3-week cycle, in which the anti-MICA / B antibody is administered once every 3 weeks (Q3W). In some embodiments, the administration interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times. In some embodiments, the repetition of the dosing interval is performed over a period of at least 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 3 years, 4 years, or 5 years.

[0097] In some embodiments of the fifth aspect, the anti-MICA / B antibody includes a light chain variable region (VL) comprising the light chain complementarity determination region 1 (LCDR1) of SEQ ID NO: 1, the light chain complementarity determination region (LCDR2) of SEQ ID NO: 2, and the light chain complementarity determination region 3 (LCDR3) of SEQ ID NO: 3, and a heavy chain variable region (VH) comprising the heavy chain complementarity determination region 1 (HCDR1) of SEQ ID NO: 4, the heavy chain complementarity determination region 2 (HCDR2) of SEQ ID NO: 5, and the heavy chain complementarity determination region 3 (HCDR3) of SEQ ID NO: 6. In some embodiments, VL comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, VH comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the light chain includes the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the heavy chain includes the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

[0098] In some embodiments of the fifth aspect, the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the anti-MICA / B antibody binds to the alpha-3 domain of the MICA protein, the MICB protein, or both the MICA protein and the MICB protein. In some embodiments, the MICA protein is a membrane-bound MICA protein, a soluble MICA protein, or both. In some embodiments, the MICB protein is a membrane-bound MICB protein, a soluble MICB protein, or both.

[0099] In some embodiments of the fifth aspect, the anti-MICA / B antibody is selected from whole immunoglobulin, scFv, Fab, F(ab')2, or disulfide-linked Fv. In some embodiments, the anti-MICA / B antibody is IgG or IgM. In some embodiments, the anti-MICA / B antibody is a humanized or chimeric antibody.

[0100] In some embodiments of the fifth aspect, the method further comprises administering an effective amount of a PD-1 inhibitor to a subject. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0101] One aspect of the present disclosure is a method for treating multiple myeloma in a subject, comprising administering to the subject a therapeutically effective dose of an antibody that binds to MICA / B (an anti-MICA / B antibody).

[0102] In some embodiments, the method further includes administering a therapeutically effective dose of an immunomodulator to the subject. In some embodiments, the method further includes administering a therapeutically effective dose of dexamethasone to the subject.

[0103] In some embodiments, the subject shows a response (e.g., complete or partial response) or disease stabilization after administration of an anti-MICA / B antibody. In some embodiments, the subject has relapsed / refractory multiple myeloma. In some embodiments, the subject is unresponsive to corticosteroids, melphalan, or a combination of vincristine, doxorubicin, and dexamethasone.

[0104] In some embodiments, the method further includes administering a therapeutically effective dose of corticosteroid to the subject prior to the first dose of anti-MICA / B antibody. In some embodiments, the corticosteroid is administered 30 to 60 minutes before the administration of anti-MICA / B antibody. In some embodiments, the corticosteroid is administered in a dose of approximately 2 mg to approximately 50 mg. In some embodiments, the corticosteroid is administered in a dose of approximately 10 mg. In some embodiments, the corticosteroid is administered orally or intravenously.

[0105] In some embodiments, the method further includes administering one or more subsequent doses of the anti-MICA / B antibody. In some embodiments, a corticosteroid is administered before the initial dose of the anti-MICA / B antibody, but not before one or more subsequent doses of the anti-MICA / B antibody. In some embodiments, the corticosteroid is dexamethasone.

[0106] This patent or application document includes at least one drawing drawn in color. Copies of this patent or patent application publication, including the color drawing(s), will be provided by the Japan Patent Office upon request and payment of the required fees.

[0107] An understanding of the features and advantages of the present invention will be obtained by referring to the following embodiments for carrying out the invention and the accompanying drawings, which describe exemplary embodiments in which the principles of the present invention are utilized.

[0108] [Brief explanation of the drawing]

[0109] [Figure 1A] The binding characteristics of CLN-619 are shown. The average of three independent experiments measuring CLN-619 binding to representative MICA allele variants and standard MICB allele variants, as measured by Octet, is presented. [Figure 1B]The binding characteristics of CLN-619 are shown. The average of two independent experiments measuring CLN-619 binding to representative MICA allele variants and standard MICB allele variants by ELISA is shown (N=2). CLN-619 exhibits comparable levels of binding to various proteins, and therefore, not all symbols are visible. [Figure 1C] This shows the binding characteristics of CLN-619. CLN-619 binding to 28 of the most common MICA allele variants, as measured by Luminex, in two independent experiments. 6D4 was used as a positive control (PMID: 28334733). Error bars represent SEM results.

[0110] [Figure 2] This image shows the CLN-619 epitope. It is an X-ray crystal structure of the human Fab antibody fragment CLN-619, complexed with the antigen MICA1 alpha-3 domain, at a resolution of 2.12 Å. The binding interface of CLN-619 is a discontinuous epitope containing 19 amino acid residues present only in the alpha-3 domain of MICA.

[0111] [Figure 3] This study demonstrates that CLN-619 modulates cell surface MICA / B through inhibition of shedding. A is a graph showing cell surface MICA / B levels measured by flow cytometry. B is a graph showing soluble MICA / B in the cell supernatant measured by ELISA (N=3 or N=2, error bars represent SEM). Geometric mean EC50 values ​​and 95% confidence intervals are reported.

[0112] [Figure 4]This shows that CLN-619 mediates NK activation and cell death of MICA / B-expressing target cells. A is a graph from an ELISA used to measure IFNγ in the cell supernatant recovered from a 96-hour co-culture death assay of NK cells and MICA / B-expressing target cells (E:T 40:1) in the presence of stepwise titrated antibodies. N=3. CLN-619 DANA is the Fc-silencing form of the mutated (D265A / N297A) CLN-619. B is a graph from the xCELLigence platform used to measure MICA / B-expressing target cell death from the experiment described in Figure 4A. A representative curve is shown.

[0113] [Figure 5] This study demonstrates that CLN-619 induces ADCC. MICA-expressing target cells treated with stepwise titrated antibodies were co-cultured with NFAT-luciferase Jurkat cells expressing human FcγRIIIa(CD16A) high-affinity (V158) or low-affinity (F158) variants in an E:T ratio of 10:1. Luciferase activity was quantified after 6 hours. Geometric mean EC50 values ​​and 95% confidence intervals are reported. E:T control background was subtracted.

[0114] [Figure 6] This study demonstrates that CLN-619 promotes antibody-dependent phagocytosis. Macrophage phagocytosis of fluorescently labeled target cells was measured by flow cytometry. Representative curves of HCC1534 and HCT-116 cells treated with CLN-619 from two donors are shown. Cetuximab and isotype control antibody treatments were performed at the highest concentrations. Geometric mean EC50 values ​​and 95% confidence intervals are reported. E:T control background was subtracted. Error bars represent SEM.

[0115] [Figure 7A]This study demonstrates that CLN-619 treatment elicits a potent antitumor effect in a preclinical mouse model. The graph shows the results for BALB / c SCID mice inoculated with PLC / PRF / 5 human hepatocellular carcinoma cells (N=10 / group). Mice were treated with ip three times weekly using two dose levels of CLN-619. Statistical analysis represents one-way ANOVA with multiple comparisons. *P=0.015. [Figure 7B] This study demonstrates that CLN-619 treatment elicits a potent antitumor effect in a preclinical mouse model. The graph shows the results for BALB / c SCID mice inoculated with HCC1534 human lung cancer cells (N=10 / group). Mice were treated twice weekly with ip (infusion therapy) using a range of CLN-619 doses or Fc-silencing CLN-619. Statistical analysis represents one-way ANOVA with multiple comparisons. NS (Not Significant) is not statistically significant. [Figure 7C] This study demonstrates that CLN-619 treatment elicits a potent antitumor effect in a preclinical mouse model. The graph shows the results for BALB / c SCID mice inoculated with HCT-116 luciferase-tagged human colon cancer cells (N=10 / group). Mice were treated twice weekly with either CLN-619 or hIgG1. Survival analysis was performed on day 41 using the Kaplan-Meier assay.

[0116] [Figure 8] This table shows the allele frequencies of MICA allele variants tested in the Luminex assay. Allele frequencies are based on high-throughput sequencing of over 2 million donor samples previously described

[16] . MICA*009 is indistinguishable from MICA*049, and MICA*001 is the standard allele of MICA.

[0117] [Figure 9]This table shows the binding of CLN-619 to FcR. The kinetic binding parameters were determined by Biacore-SPR. Biacore chips were coupled to various his-tagged FcR via anti-HIS antibodies, and the test antibody was passed through the bound FcR. Kinetic analysis was performed using Biacore software. Relative affinity scale (M): ++++ 10⁻⁸ to -⁹, +++ 10⁻⁷, ++ 10⁻⁶, + 10⁻⁵, + / - detectable binding, - undetectable binding.

[0118] [Figure 10] This table shows the sequence analysis of the CLN-619 epitope of the MICA / B allele. The alleles listed are those with a frequency of over 1% in the population and exhibit high homology with conserved amino acid changes, indicated in orange. * indicates a standard allele.

[0119] [Figure 11] This graph shows that CLN-619 does not interfere with MICA-NKG2D binding. Primary NK cells were incubated with His-tagged MICA ECD in the presence of CLN-619 or hIgG1 control antibody, either without (a) or with anti-NKG2D antibody. The amount of MICA bound to the cell surface was measured by flow cytometry.

[0120] [Figure 12] This graph shows that CLN-619 DANA retains the ability to regulate cell surface MICA / B. Cell surface levels of MICA / B were measured by flow cytometry.

[0121] [Figure 13A] This shows the pharmacokinetic parameters of CLN-619 after a single intracellular injection in BALB / c SCID mice. A is a graph showing the concentration-time profiles of CLN-619 after single intracellular injections of 1, 3, and 10 mg / kg, and B shows the pharmacokinetic parameters of CLN-619 after single intracellular injections of 1, 3, and 10 mg / kg. The mean and standard deviation are reported. [Figure 13B] This shows the pharmacokinetic parameters of CLN-619 after a single intracellular injection in BALB / c SCID mice. A is a graph showing the concentration-time profiles of CLN-619 after single intracellular injections of 1, 3, and 10 mg / kg, and B shows the pharmacokinetic parameters of CLN-619 after single intracellular injections of 1, 3, and 10 mg / kg. The mean and standard deviation are reported.

[0122] [Figure 14] This table shows the baseline characteristics of patients enrolled in a phase 1 open-label dose-escalation study of CLN-619 for the treatment of advanced solid tumors.

[0123] [Figure 15] This is a schematic diagram showing the structure of dose-escalation studies, including the number of patients in each dose-level cohort.

[0124] [Figure 16] This table summarizes the treatment-induced adverse events (TEAEs) observed in 10% or more of patients.

[0125] [Figure 17A] These graphs and tables show the mean serum concentration and PK parameters of CLN-619 at various time points after administration, stratified by dosage level. [Figure 17B] These graphs and tables show the mean serum concentration and PK parameters of CLN-619 at various time points after administration, stratified by dosage level.

[0126] [Figure 18] These are a series of graphs showing cytokine expression in longitudinal serum samples from patients treated with CLN-619 monotherapy, tested on the Luminex platform using Myriad RBM Inflammation and custom MAP. The graphs show the mean absolute cytokine level (pg / ml) per dose level over time. Error bars represent standard deviation (SD).

[0127] [Figure 19A] This graph shows the duration of treatment and clinical activity for each study patient who received CLN-619 monotherapy. [Figure 19B] This graph shows the duration of treatment and clinical activity for each study patient who received CLN-619 monotherapy.

[0128] [Figure 20] This table summarizes the objective response and best monotherapy effect in study patients who received CLN-619 monotherapy.

[0129] [Figure 21] These are a series of images of a research patient with mucoepidermoid carcinoma of the parotid gland at baseline (top panel), during the fourth cycle of CLN-619 treatment (middle panel), and during the seventh cycle of CLN-619 treatment (bottom panel).

[0130] [Figure 22] These are images of a research patient with endometrial cancer at baseline (left panel) and after 10 cycles of CLN-619 treatment (right panel).

[0131] [Figure 23A] This document outlines the overall study design for clinical trials of CLN-619, both as monotherapy and in combination with pembrolizumab, in patients with advanced solid tumors. [Figure 23B] This document outlines the overall study design for clinical trials of CLN-619, both as monotherapy and in combination with pembrolizumab, in patients with advanced solid tumors.

[0132] [Figure 24] This document presents the event schedule for clinical studies of CLN-619, both as monotherapy and in combination with pembrolizumab, in patients with advanced solid tumors.

[0133] [Figure 25]This shows the sampling schedule for clinical studies of CLN-619, both as monotherapy and in combination with pembrolizumab, in patients with advanced solid tumors.

[0134] [Figure 26A] CLN-619-001 indicates the infusion time.

[0135] [Figure 26B] This shows the CLN-619-001 infusion time instructions for Module D (loading dose cohort).

[0136] [Figure 27] This shows the duration of treatment and clinical activity in the combined cohort.

[0137] [Figure 28] This shows the duration of treatment and clinical activity in patients with NSCLC.

[0138] [Figure 29] This study shows the duration of treatment and clinical activity in cohorts receiving monotherapy at doses of 1 mg / kg or higher.

[0139] [Figure 30] This is an overview of treatment-related adverse events (TEAEs).

[0140] [Figure 31] This paper presents the overall study design for the use of CLN-619 for the treatment of R / R multiple myeloma.

[0141] [Figure 32A] This graph shows the effect of CLN-619 on T cell activation. It is a graph from the xCELLigence platform used to measure cell lysis in a 72-hour co-culture death assay combining negatively selected CD8+ T cells from PBMCs with MICA / B-expressing target cells (E:T 10:1) in the presence of stepwise titrated antibodies. N=3. A representative curve is shown. [Figure 32B]This shows the effect of CLN-619 on T cell activation. The graph is from flow cytometry used to measure the percentage of CD8+ T cells expressing CD25 from the experiment described in Figure 32A. N=3. A representative curve is shown. [Figure 32C] This shows the effect of CLN-619 on T cell-mediated cell lysis. IFNγ was measured in the cell supernatant collected from the experiment described in Figure 32A using ELISA. N=3. A representative curve is shown. [Modes for carrying out the invention]

[0142] In some embodiments, monoclonal antibodies that specifically bind to MICA / B are disclosed herein. In some embodiments, the MICA / B antibodies herein bind to the MICA / B protein or a fragment thereof, modulating the immune response of an individual and thereby treating cancer (e.g., hepatocellular carcinoma).

[0143] Major histocompatibility complex class I-associated chains A and B (MICA / B) are two stress-inducible ligands for the natural killer (NK) receptor NKG2D and play a crucial role in mediating NK and T cell cytotoxicity. Soluble MICA / B, shed by affected cells (e.g., cancer cells), desensitizes NK and T cells via binding to the NKG2D receptor, thereby suppressing the immune response. Therefore, regulation of MICA / B is useful for modulating the immune response in individuals, such as those with cancer.

[0144] The development of therapeutic agents that inhibit MICA / B shedding represents a novel strategy leveraging these NKG2D ligands. Clinical data demonstrating the clinical safety and efficacy of targeted inhibition of MICA / B shedding using CLN-619, a humanized IgG1 antibody that is currently the only MICA / B-targeting mAb in clinical practice, are disclosed herein. CLN-619 prevents shedding of cancer cell-derived MICA / B by binding to its alpha-3 domain and also contains an active Fcγ1 domain that drives ADCC and ADCP. Despite the high polymorphism of the MICA / B gene, CLN-619 showed surprisingly broad responsiveness to all allele variants tested. Treatment with CLN-619 resulted in a decrease in the levels of shedding MICA and an associated increase in MICA on the surface of tumor cells. Importantly, the alpha-1 and alpha-2 domains of MICA that bind to NKG2D are not inhibited by the bound antibody. In fact, CLN-619 enhances the binding of MICA to NKG2D, which is thought to depend on the antibody's Fcγ1 domain, which can simultaneously bind to CD16A on NK cells. Notably, CLN-619 exhibits attractive monotherapy activity at low doses in tumor xenograft models, given its activity heavily reliant on the functional Fcγ1 domain. Therefore, CLN-619 shows potential therapeutic efficacy as a monotherapy or in combination with other agents that bind to other NK cell receptors and / or one or more checkpoint inhibitors.

[0145] Stabilization of MICA / B by CLN-619 leads to the accumulation of these NKG2D ligands on the tumor cell surface, thereby overcoming immune evasion through MICA / B shedding. While not bound by any specific theory or mechanism of action, this likely maximizes the ADCC and ADCP function of antibodies due to increased target antigen density. Importantly, CLN-619 has the potential to activate NK cells through the simultaneous, and possibly synergistic, binding of two key activating receptors, NKG2D and CD16A. The synergistic action of the two receptors is supported by published data showing that NKG2D binding can lower the activation threshold of several receptors, including CD16A. Given that NK cell activating receptors in TMEs are often downregulated, the synergistic action of the two receptors may be significant. The disclosures herein include key data from a clinical trial of CLN-619 (Phase 1 clinical trial number NCT05117476) in patients with advanced solid tumors treated with CLN-619 alone or in combination with pembrolizumab.

[0146] Monotherapy activity is not common in Phase 1 oncology studies, as evidenced by retrospective analyses showing low single-digit objective response rates among more than 4,000 patients enrolled in Phase 1 solid tumor studies funded by the National Cancer Institute between 2000 and 2019. The monotherapy response rate of CLN-619 in evaluable patients is superior to both past NCI data and TIGIT and lag-3 antibodies, which truly supports this in the early stages of the CLN-619 program.

[0147] Further details of clinical trials of CLN-619 in patients with advanced solid tumors treated with CLN-619 alone or in combination with pembrolizumab can be found in the appendix.

[0148] One such study is a Phase 1 dose-escalation and expansion study testing both CLN-619 monotherapy and CLN-619 in combination with pembrolizumab in a staggered parallel cohort. CLN-619 monotherapy was administered every three weeks. Premedication was required only before the first infusion and was mandated to include a corticosteroid starting at a 3 mg / mg dose level. Dose-limiting toxicity was assessed during the first cycle, and response was assessed every three cycles according to RECIST criteria.

[0149] The monotherapy group used a standard 3x3 dose escalation study dose of 0.1–10 mg / mg. Dose levels cleared for dose levels (DLTs) could be further expanded (sometimes referred to as "backfill") until a total of up to 10 patients were treated per dose level cleared. A total of 37 patients were treated (median patient age 63, 62% were female). This group, having received multiple prior treatments, had a median of 3 lines of prior treatment, and slightly more than half had experienced CPI prior treatment.

[0150] RECIST responses were observed at doses starting from 3 mg / kg, including an ongoing sustained complete response (CR) in a parotid gland patient treated with 3 mg / kg, and two partial responses (PR) in endometrial patients treated with 3 and 10 mg / kg. Both the parotid CR and one of the endometrial PRs occurred in patients whose disease had progressed during prior checkpoint inhibitor treatment. At doses of 1 mg / kg or higher, 10 out of 22 evaluable patients achieved the best objective response or disease stabilization.

[0151] In addition to two endometrial partial responses (PR), three cervical cancer patients achieved disease stabilization, and two ovarian cancer patients also achieved disease stabilization. The data show that seven of the ten gynecological cancer patients enrolled in the study achieved the best possible outcome, either an objective response or disease stabilization.

[0152] Sustained disease stabilization was also observed in one breast cancer patient and one salivary gland cancer patient, highlighting the objective clinical activity of monotherapy observed across diverse tumor types.

[0153] In scans of a patient with persistent complete response (CR) and parotid gland tumor, biopsy-proven lymph node recurrence met the criteria for complete response at C4D1, was confirmed at C7D1, and was ongoing at the data cutoff more than 9 months after the initiation of treatment with CLN-619. Notably, this patient had experienced a 30-month sustained partial response (PR) to checkpoint inhibitor therapy prior to progression during treatment.

[0154] In a scan from one of two endometrial patients who experienced a confirmed partial response (PR), baseline images showed multiple bilateral lung metastases that had disappeared by C4D1, meeting the criteria for a PR, and were confirmed by subsequent scans at C7D1. This patient had progressed after prior treatment with platinum-based chemotherapy plus Herceptin, anastrazole, doxorubicin, and most recently, pembrolizumab plus lenvatinib for 9 months.

[0155] The fact that more than 10% of patients experienced adverse events under treatment demonstrates that the majority of AEs observed during CLN-619 monotherapy were grade 1 / 2. Notably, no dose-limiting toxicities were observed at any of the dose levels evaluated. A single grade 3 treatment-related AE of laryngeal edema, one of the infusion-associated adverse events observed with other monoclonal antibodies, occurred during the first infusion in a patient who did not receive the mandatory steroid premedication. All other IRRs were grade 1 or 2 and occurred only during the first cycle. The data support the good tolerability of CLN-619 monotherapy, and therefore the safety profile should support its ease of use in combination with other drugs.

[0156] The data further demonstrate that CLN-619 monotherapy was well-tolerated up to 10 mg / kg without dose-limiting trials (DLTs), and that CLN-619 showed remarkable monotherapy activity, including confirmed and ongoing objective responses, across a variety of tumor types, including patients whose disease progressed during checkpoint inhibitor treatment.

[0157] Considering the widespread tumor expression along the pathway, it is likely that CLN-619 has pan-cancer therapeutic activity.

[0158] The data presented in the examples herein demonstrate the safety and efficacy of CLN-619 monotherapy, showing acceptable safety and promising clinical activity, including objective responses in multiple tumor types, in both checkpoint-experienced and checkpoint-inexperienced patients. Of particular note is the remarkable monotherapy activity observed in multiple gynecological malignancies, such as endometrial cancer and cervical cancer. This clinical activity in multiple gynecological cancer types demonstrates the potential of CLN-619 in patients with high unmet needs. More specifically, monotherapy dose escalation demonstrated an acceptable safety profile of CLN-619 across all evaluated doses (0.1, 0.3, 1, 3, 6, and 10 mg / kg), and no dose-limiting toxicities were observed. Consistent with other therapeutic monoclonal antibodies, all infusion-associated reactions were grade 1 / 2 and were limited to the first dose in patients who had received mandatory premedication. Importantly, CLN-619 therapy was effective against multiple tumor types at the administered dose, demonstrating disease stabilization in different patient tumors, including cervical cancer, ovarian cancer, salivary gland cancer, and breast cancer. Certain patients had undergone multiple prior treatments, with a median of 3 prior systemic treatments (1–7), and 54% had previously received immune checkpoint inhibitors. The initial clinical data on CLN-619 monotherapy demonstrate broad potential across diverse tumor types. Therefore, CLN-619 has the potential to treat a critical class of patients whose tumors have relapsed or were unsuitable for checkpoint inhibitor therapy.

[0159] Further data indicate that combination therapy with CLN-619 and pembrolizumab was well-tolerated and demonstrated meaningful clinical activity, including objective responses, in tumor types typically refractory to pembrolizumab.

[0160] Data from CLN-619-001 (NCT05117476), a first-in-human dose-escalation and dose-expansion study of CLN-619 alone or in combination with anti-PD1 in patients with advanced solid tumors, are described in the examples. Sixty-four patients received CLN-619 in combination with pembrolizumab (n=22) or CLN-619 as monotherapy (n=42).

[0161] Monotherapy efficacy, including confirmed responses, was observed at doses of 3 mg / kg or higher across several tumors. CLN-619 was tolerable at doses up to 10 mg / kg, and no dose-limiting toxicities were observed at any dose tested. In some embodiments, CLN-619 monotherapy was administered intravenously every three weeks.

[0162] Pharmacokinetic (PK) studies, pharmacodynamic (PD) studies, and mechanism of action studies were conducted with the aim of observing the PK and PD effects of CLN-619 in the peripheral and tumor microenvironments.

[0163] PK analysis of dose escalation with CLN-619 monotherapy showed that the half-life of CLN-619 ranged from 61.3 to 443 hours (2.5 to 18.45 days) for dose levels ranging from 0.1 to 10 mg / kg. A nearly dose-proportional increase in exposure, measured by Cmax, was observed at 0.1 to 10 mg / kg. In addition, a greater-than-dose-proportional increase in CLN-619 exposure, measured by AUC0-504h, was observed at 0.1 to 1 mg / kg, while a nearly dose-proportional increase in CLN-619 exposure was observed at dose levels of 1 to 10 mg / kg.

[0164] Cytokine expression in longitudinal serum samples from patients treated with CLN-619 monotherapy was investigated. The data showed a transient increase in cytokine levels observed 2–6 hours after the first dose of CLN-619, decreasing to baseline by days 4–8. No dose-dependent increase in cytokine secretion was observed in the periphery.

[0165] Increased MICA / B expression was observed within the tumor at the time of treatment. Analysis of nine paired biopsies using multiple IF and quantification using the image analysis algorithm (Flagship Biosciences) showed increased MICA / B positive staining in the majority of paired biopsies from patients during treatment (2nd cycle, day 8) compared to baseline.

[0166] MICA / B membrane localization in nine paired biopsies was also analyzed by IHC and quantified as the percentage of membrane-stained cells by pathology, showing an increase in MICA / B membrane positivity in the majority of biopsies during treatment.

[0167] Data in the addendum demonstrate that monotherapy activity was achieved in patients who had received multiple prior treatments, including those with progression in anti-PD1 therapy. Objective responses were observed with CLN-619 monotherapy across multiple tumor types, including those with progression in checkpoint inhibitor therapy, and noteworthy monotherapy activity was observed in gynecological malignancies.

[0168] The data in the examples also present two case studies of subjects with endometrial tumors. In the first case study, the subject had endometrioid endometrial carcinoma and demonstrated a confirmed partial response after 3 cycles of CLN-619. The subject had received 3 lines of prior treatment and had not received checkpoint inhibitor intervention. In the second case study, the subject had serous endometrial carcinoma and demonstrated a confirmed partial response after 6 cycles of treatment with CLN-619. This subject had received 5 lines of prior treatment, including pembrolizumab and lenvatinib. The data indicate that the responding endometrial tumors are microsatellite stable and have low tumor mutational load and low novel antigen presentation index.

[0169] The tumor microenvironment in patients with disease stability lasting more than 6 months was analyzed using paired biopsies from 13 patients treated in dose-level extension cohorts at dose levels of 3, 6, and 10 mg / kg. The data showed low tumor mutational load and low novel antigen presentation in tumors with long-term disease stability. Increased MICA / B expression and NK cell activation were observed during treatment with CLN-619 in tumors from patients with disease stability lasting more than 6 months. PDL1 expression and C8 activation increased during treatment in 2 / 3 of the paired biopsies.

[0170] The study also demonstrates that soluble MICA (sMICA) in patient serum is a PD biomarker for CLN-619. In this study, endogenous IgG was pre-removed from negative serum samples or normal serum supplemented with recombinant sMICA using Protein G Sepharose beads. The supernatant after bead removal was immunoprecipitated with hIgG1 or CLN-619. The sMICA remaining in the supernatant after immunoprecipitation was quantified by ELISA and plotted as a percentage relative to control. The study shows that CLN-619 binds to sMICA in human serum. It was further shown that serum sMICA levels increased upon CLN-619 treatment, and that sMICA levels act as a surrogate for target binding by CLN-619. This is demonstrated by modeling CLN-619 drug concentrations that predicted sMICA levels. Furthermore, no sink effect was observed. CLN-619 was found in patient blood at approximately 1000 times higher levels compared to sMICA, suggesting that only a small fraction of CLN-619 is consistently bound to sMICA. This small fraction of CLN-619 bound to sMICA was not found to be clinically relevant.

[0171] Therefore, consistent with the proposed mechanism of action of CLN-619, an increase in the membrane localization of MICA / B was observed during treatment with CLN-619. Accordingly, the present invention presents a method for increasing the localization of MICA / B in a subject by administering an effective amount of CLN-619 to that subject.

[0172] In certain embodiments, the presence of sMICA is shown to be a pharmacodynamic biomarker of CLN-619. The method of the present invention comprises monitoring the level of sMICA in the presence and absence of CLN-619 therapy, where an increase in the presence of sMICA indicates that CLN-619 is therapeutically effective. The data herein show that the level of increase in sMICA is dose-responsive and dependent on the dose of CLN-619 administered.

[0173] Other methods described herein use sMICA as a measure of target binding by CLN-619. These methods, for example, are methods for determining the potency or effectiveness of CLN-619 in a subject, comprising determining the presence of sMICA in the subject's serum before and after administration of CLN-619, wherein an increase in the presence of sMICA indicates the potency or effectiveness.

[0174] Furthermore, the data showing objective response and long-term disease stabilization (more than 6 months) in tumors with low TMB and low antigen-presenting index support the possibility that CLN-619 can induce a response without relying on a sophisticated antigen-presenting mechanism. This is consistent with the diverse proposed mechanisms of action of CLN-619. Accordingly, a method for treating a solid tumor with CLN-619 is described herein, wherein the treatment results in at least a partial response of the tumor to CLN-619. The method may also include showing disease slowing or stabilization over a period of at least 6 months. Accordingly, the method of the present invention may also be used to extend the lifespan of a subject for a period of at least 6 months using the composition of the present invention. These and other aspects of the present invention are described in further detail below herein.

[0175] definition

[0176] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this disclosure pertains.

[0177] As used herein, "MICA / B" refers to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein, including their variants, isoforms, and species homologs of human MICA / B.

[0178] As used herein, “antibody” refers to a glycoprotein that exhibits binding specificity to a particular antigen. Antibodies often contain a variable domain and a constant domain in both the heavy chain and the light chain. Therefore, most antibodies have a heavy chain variable domain (VH) and a light chain variable domain (VL), which together form the portion of the antibody that binds to the antigen. Each variable domain has three complementarity-determining regions (CDRs) that form a loop in the heavy chain variable domain (VH) and the light chain variable domain (VL) and come into contact with the surface of the antigen. As used herein, antibodies include complete molecules and their functional fragments, the functional fragments also referred to as “antigen-binding portions” or fragments of the antibody that can bind to an antigen.

[0179] As used herein, a “chimeric” antibody is an antibody that, insofar as it exhibits the desired biological activity, has a portion of its heavy and / or light chain that is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, and the remainder of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, and in a fragment of such an antibody (see, for example, Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)). As used herein, a “humanized antibody” refers to a chimeric antibody in which a human sequence is substituted in the antibody sequence.

[0180] The terms “recipient,” “individual,” “subject,” “host,” and “patient” are used interchangeably herein and, as may be, refer to any mammalian subject, in particular human, to whom diagnosis, treatment, or therapy is desired. “Mammal” for treatment purposes means any animal classified as a mammal, e.g., humans, domesticated and livestock animals, as well as experimental, zoo, sporting, or companion animals, e.g., dogs, horses, cats, cattle, sheep, goats, pigs, mice, rats, rabbits, guinea pigs, monkeys, etc. In some embodiments, the mammal is human.

[0181] As used herein, terms such as “treatment” and “to treat” may, in some cases, mean administering a drug or performing a procedure for the purpose of obtaining an effect. Such effect may be preventive in that it completely or partially prevents a disease or its symptoms, and / or therapeutic in that it results in a partial or complete cure of the disease and / or its symptoms. “Treatment” as used herein may include treatment of a disease or disorder (e.g., cancer) in mammals, in particular humans, and may include (a) preventing the development of the disease or its symptoms in a subject that is considered susceptible to the disease but has not yet been diagnosed as having the disease (e.g., a disease that may be associated with or may be caused by the underlying disease), (b) inhibiting the disease, i.e., stopping its development, and (c) alleviating the disease, i.e., causing the disease to disappear. Treating means a reduction, remission, decrease in symptoms or making the disease state more tolerable to the patient, a slowing of the rate of degeneration or decline, or the cessation of degeneration. The term "treatment" may refer to any indication of treatment success or improvement or prevention of cancer, including any objective or subjective parameters, such as making a point less debilitating. Treatment or improvement of symptoms is based on one or more objective or subjective parameters, including the results of a physician's examination. Accordingly, the term "treatment" includes the administration of the compounds or agents of the present invention to prevent or delay the onset of symptoms or conditions associated with a disease (e.g., cancer), reduce such symptoms or conditions, or halt or inhibit the onset of such symptoms or conditions. The term "therapeutic effect" refers to the reduction, elimination, or prevention of a disease, symptoms of a disease, or side effects of a disease in a subject.

[0182] "Therapeutic dose" sometimes refers to an amount sufficient to treat a disease when administered to a subject in order to treat that disease.

[0183] As used herein, the singular forms "a," "and," and "the" refer to multiple objects unless otherwise clearly indicated by the context. Therefore, for example, a reference to "an antibody" includes multiple antibodies.

[0184] When used herein, a number preceded by "approximately" refers to a range that includes that number and extends from a value 10% less than that number to a value 10% greater than that number. A range preceded by "approximately" refers to a range that extends from a value 10% less than the lower limit of that range to a value 10% greater than the upper limit of that range.

[0185] "Identical percentage (%)" refers to the degree to which two sequences (nucleotides or amino acids) have the same residues at the same positions in their alignment. For example, "The amino acid sequence is X% identical to sequence number Y" refers to the percentage of identity of the amino acid sequence with respect to sequence number Y, stating that X% of the residues in the amino acid sequence are identical to the residues in the sequence disclosed in sequence number Y. Generally, computer programs are used for such calculations. Exemplary programs for comparing and aligning pairs of sequences include ALIGN (Myers and Miller, 1988), FASTA (Pearson and Lipman, 1988, Pearson, 1990), and gapped BLAST (Altschul et al., 1997), BLASTP, BLASTN, or GCG (Devereux et al., 1984).

[0186] MICA / B

[0187] In some embodiments, monoclonal antibodies that specifically bind to MICA / B are disclosed herein. In some embodiments, monoclonal antibodies that competitively bind to MICA / B are further disclosed herein.

[0188] The major histocompatibility complex (MHC) class I chain-associated gene A and gene B proteins (MICA / B) are glycosylated, polymorphic, membrane-anchored, non-classical MHC class I proteins. MICA / B are associated with MHC class I and possess a similar domain structure including three extracellular Ig-like domains (alpha-1, alpha-2, and alpha-3), a transmembrane domain, and a C-terminal cytoplasmic tail. MICA / B are ligands for natural killer group 2D (NKG2D), a type C lectin-like activating receptor in immune effector cells, including NK cells, NKT cells, and both αβ and γδ CD8+ T cells. The interaction between MICA / B and NKG2D plays a role in tumor surveillance and immune responses.

[0189] MICA / B proteins are typically expressed at low levels in normal cells but are induced to higher levels in stressed or transformed cells (e.g., cancer cells). Interaction between NKG2D-carrying immune effector cells and stressed or diseased cells expressing MICA / B ligands on their cell surface results in a cellular immune response against the stressed / disease cells, ultimately leading to the death of MICA / B-expressing cells. In cancer cells, cleaved MICA / B proteins (proteins lacking a transmembrane domain and cytoplasmic tail but retaining three extracellular domains including alpha-1, -2, and -3 domains) are often shedged into the bloodstream by protease action, leading to downregulation (receptor internalization) of NKG2D, their intended receptor, in effector immune cells. In some cases, MICA / B glycoproteins are not always destined to be produced intracellularly and bound to the cell surface membrane; instead, they are incorporated into exosomes and released extracellularly, where they interact with the NKG2D receptor in immune cells. These cleaved or soluble MICA / B ligands, shed from the surface of cancer cells, function like decoy molecules, causing downregulation of the NKG2D receptor in immune effector cells such as NK, NKT, and various CD8+ T cells. In some cases, the formation of soluble MICA / B leads to an abnormal situation where effectors of the natural defense system, whose original role is to seek out and destroy transformed cells, are inactivated by the immunosuppressive effects of these decoy ligand molecules, thereby allowing cancer cells to hide from the immune system and grow unsuppressed.

[0190] Anti-MICA / B antibody

[0191] Antibodies that specifically bind to the MICA / B protein are provided herein. In some embodiments, the anti-MICA / B antibody comprises at least one heavy chain and at least one light chain. In some embodiments, the anti-MICA / B antibody comprises at least one heavy chain containing a heavy chain variable domain (VH) and at least one light chain containing a light chain variable domain (VL). Each VH and VL contains three complementarity-determining regions (CDRs). The amino acid sequences of the VH and VL and the CDRs determine the antigen-binding specificity and antigen-binding strength of the antibody. The amino acid sequences of the heavy chain and light chain, VH and VL and the CDRs are summarized in Table 1.

[0192] [Table 1-1] [Table 1-2]

[0193] In some embodiments, the antibody specifically binds to the MICA protein. In some embodiments, the antibody specifically binds to the MICB protein. In some embodiments, the antibody specifically binds to both the MICA and MICB proteins. In some embodiments, the antibody specifically binds to the alpha-3 domain of the MICA protein. In some embodiments, the antibody specifically binds to the alpha-3 domain of the MICB protein. In some embodiments, the antibody specifically binds to the alpha-3 domains of both the MICA and MICB proteins. In some embodiments, the antibody binds to the MICA protein, which is a membrane-bound MICA protein. In some embodiments, the antibody binds to the MICA protein, which is a soluble MICA protein. In some embodiments, the antibody binds to the MICA protein, which is both a membrane-bound and soluble MICA protein. In some embodiments, the antibody binds to the MICB protein, which is a membrane-bound MICB protein. In some embodiments, the antibody binds to the MICB protein, which is a soluble MICB protein. In some embodiments, the antibody binds to the MICB protein, which is both a membrane-bound and soluble MICB protein.

[0194] In some embodiments, the antibody that specifically binds to MICA / B is a monoclonal antibody. In some embodiments, the antibody is an antigen-binding fragment. In some embodiments, the antibody is selected from whole immunoglobulin, scFv, Fab, F(ab')2, or disulfide-bonded Fv. In some embodiments, the antibody is IgG or IgM. In some embodiments, the antibody is humanized. In some embodiments, the antibody is chimeric.

[0195] MICA / B antibody complementation determination region

[0196] Antibodies that specifically bind to MICA / B having a light chain containing a light chain complementarity-determining region (CDR) are disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a light chain CDR sequence having at least about 70% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 1-3. In some embodiments, the antibody that binds to MICA / B includes a light chain CDR sequence having at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 1-3. In some embodiments, the antibody that binds to MICA / B includes a light chain CDR sequence having 100% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 1-3.

[0197] Antibodies that specifically bind to MICA / B having a heavy chain containing a heavy chain complementarity-determining region (CDR) are further disclosed herein. In some embodiments, the antibody that binds to MICA / B comprises a heavy chain CDR sequence having at least about 70% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 4-6. In some embodiments, the antibody that binds to MICA / B comprises a heavy chain CDR sequence having at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 4-6. In some embodiments, the antibody that binds to MICA / B comprises a heavy chain CDR sequence having 100% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 4-6.

[0198] Antibodies that bind to MICA / B, including light chain complementarity-determining regions (CDRs) and heavy chain complementarity-determining regions (CDRs), are also disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a light chain CDR sequence having at least about 70% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 1-3, and a heavy chain CDR sequence having at least about 70% identical amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 4-6. In some embodiments, the antibody that binds to MICA / B includes a light chain CDR sequence having an amino acid sequence identical to at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 4-6, and a heavy chain CDR sequence having an amino acid sequence identical to at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence to at least one of the amino acid sequences described in SEQ ID NOs: 4-6. In some embodiments, the antibody that binds to MICA / B includes a light chain CDR sequence having an amino acid sequence that is 100% identical to at least one of the amino acid sequences described in SEQ ID NOs: 1-3, and a heavy chain CDR sequence having an amino acid sequence that is at least approximately 100% identical to at least one of the amino acid sequences described in SEQ ID NOs: 4-6.

[0199] In some embodiments, the antibody that binds to MICA / B includes at least one of the following: light chain CDR1 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 1, light chain CDR2 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 2, and light chain CDR3 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the antibody that binds to MICA / B includes light chain CDR1 having an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 1, and light chain CDR1 having an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, In some embodiments, the antibody that binds to MICA / B includes at least one of the following: a light chain CDR2 having an amino acid sequence identical to 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%; and a light chain CDR3 having an amino acid sequence identical to at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence described in SEQ ID NO: 3.

[0200] In some embodiments, the antibody that binds to MICA / B includes at least one of the following: heavy chain CDR1 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 4; heavy chain CDR2 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 5; and heavy chain CDR3 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the antibody that binds to MICA / B includes heavy chain CDR1 having an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 4; and heavy chain CDR1 having an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87% identical to the amino acid sequence described in SEQ ID NO: 5. In some embodiments, the antibody that binds to MICA / B includes at least one of the following: heavy chain CDR2 having an amino acid sequence identical to 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%; heavy chain CDR3 having an amino acid sequence identical to at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence described in SEQ ID NO: 6.

[0201] In some embodiments, the antibody that binds to MICA / B includes at least one of the following: a light chain CDR1 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 1; a light chain CDR2 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 2; a light chain CDR3 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 3; a heavy chain CDR1 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 4; a heavy chain CDR2 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 5; and a heavy chain CDR3 having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 6.In some embodiments, the antibody that binds to MICA / B has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 1, and an antibody that is at least about 75%, 80%, 81%, 82%, 83%, 84%, or 8% identical to the amino acid sequence described in SEQ ID NO: 2. Light chain CDR2 having an amino acid sequence identical by 5%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, to the amino acid sequence described in SEQ ID NO: 3, and light chain having an amino acid sequence identical by at least approximately 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the amino acid sequence described in SEQ ID NO: 3. Heavy chain CDR1 has an amino acid sequence that is at least approximately 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in CDR3, SEQ ID NO: 4. It comprises at least one of the following: a heavy chain CDR2 having an amino acid sequence identical by %, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%; and a heavy chain CDR3 having an amino acid sequence identical by at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the amino acid sequence described in SEQ ID NO: 6.In some embodiments, the antibody that binds to MICA / B includes at least one of the following: a light chain CDR1 having an amino acid sequence 100% identical to the amino acid sequence described in SEQ ID NO: 1; a light chain CDR2 having an amino acid sequence 100% identical to the amino acid sequence described in SEQ ID NO: 2; a light chain CDR3 having an amino acid sequence 100% identical to the amino acid sequence described in SEQ ID NO: 3; a heavy chain CDR1 having an amino acid sequence 100% identical to the amino acid sequence described in SEQ ID NO: 4; a heavy chain CDR2 having an amino acid sequence 100% identical to the amino acid sequence described in SEQ ID NO: 5; and a heavy chain CDR3 having an amino acid sequence 100% identical to the amino acid sequence described in SEQ ID NO: 6.

[0202] MICA / B antibody variable domain

[0203] Antibodies that specifically bind to MICA / B having a light chain containing a light chain variable domain (VL) are disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a light chain variable domain (VL) having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 7. In some embodiments, the VL has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 7. In some embodiments, the VL has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 7.

[0204] Antibodies that specifically bind to MICA / B having a heavy chain containing a heavy chain variable domain (VH) are further disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a heavy chain variable domain (VH) having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the VH has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the VH has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 8.

[0205] Antibodies that bind to MICA / B, comprising a light chain variable domain (VL) and a heavy chain variable domain (VH), are also disclosed herein. In some embodiments, the antibody that binds to MICA / B comprises a light chain variable domain (VL) having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 7 and a heavy chain variable domain (VH) having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, VL has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 7, and VH has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, VL has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 7, and VH has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 8.

[0206] MICA / B antibody heavy and light chains

[0207] Antibodies that specifically bind to MICA / B having a light chain are disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a light chain having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 9. In some embodiments, the light chain has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 9. In some embodiments, the light chain has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 9.

[0208] Antibodies that specifically bind to MICA / B having a heavy chain are further disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a heavy chain having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the heavy chain has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the heavy chain has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 10.

[0209] Antibodies that bind to MICA / B, including light and heavy chains, are also disclosed herein. In some embodiments, the antibody that binds to MICA / B includes a light chain having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 9, and a heavy chain having an amino acid sequence that is at least about 70% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the light chain has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 9, and the heavy chain has an amino acid sequence that is at least about 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the light chain has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 9, and the heavy chain has an amino acid sequence that is 100% identical to the amino acid sequence described in SEQ ID NO: 10.

[0210] Treatment and Usage Instructions

[0211] A method for treating cancer in an individual requiring treatment for cancer is provided herein, comprising the administration of an anti-MICA / B antibody disclosed herein.

[0212] In some embodiments, the antibodies disclosed herein reduce the level of soluble MICA protein. In some embodiments, the antibodies disclosed herein reduce the level of soluble MICB protein. In some embodiments, the antibodies disclosed herein reduce the levels of both soluble MICA protein and soluble MICB protein. In some embodiments, the antibodies disclosed herein suppress the shedding of soluble MICA protein. In some embodiments, the antibodies disclosed herein suppress the shedding of soluble MICB protein. In some embodiments, the antibodies disclosed herein suppress the shedding of both soluble MICA protein and soluble MICB protein. In some embodiments, the antibodies disclosed herein inhibit the shedding of soluble MICA protein. In some embodiments, the antibodies disclosed herein inhibit the shedding of soluble MICB protein. In some embodiments, the antibodies disclosed herein inhibit the shedding of both soluble MICA protein and soluble MICB protein. In some embodiments, treatment using the disclosed anti-MICA / B antibody results in the mitigation or inhibition of the immunosuppressive environment by preventing or blocking the interaction between soluble MICA / B and the NKG2D receptor in the individual.

[0213] subject

[0214] In some embodiments, the subjects described herein are subjects having cancer. The subjects may be mammals, e.g., primates, e.g., higher primates, e.g., humans (e.g., patients with the disorder described herein, e.g., cancer, or at risk of having cancer). In one embodiment, the subjects require enhancement of the immune response. In one embodiment, the subjects have the disorder described herein, e.g., cancer, or at risk of having cancer. In certain embodiments, the subjects are immunocompromised or at risk of becoming immunocompromised.

[0215] In some embodiments, the subjects being treated are those who have previously been treated with checkpoint inhibitor therapy. In some embodiments, the checkpoint inhibitor therapy includes a PD-1 inhibitor (e.g., an anti-PD-1 antibody). In some embodiments, the subjects responded to the PD-1 inhibitor (e.g., showed a complete or partial response). In some embodiments, the subjects had a response period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months or longer after administration of the PD-1 inhibitor. In some embodiments, the subjects progressed after the response period to the PD-1 inhibitor. In some embodiments, the subjects received treatment with an anti-MICA / B antibody after progression following the response period to the PD-1 inhibitor.

[0216] In some embodiments, subjects showed disease stabilization upon administration of PD-1 inhibitors. In some embodiments, subjects had disease stabilization periods of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer. In some embodiments, subjects progressed after the disease stabilization period following treatment with PD-1 inhibitors. In some embodiments, subjects received treatment with anti-MICA / B antibodies after disease progression following the disease stabilization period after treatment with PD-1 inhibitors.

[0217] In some embodiments, treatment with a PD-1 inhibitor was discontinued before the initiation of administration of an anti-MICA / B antibody. In some embodiments, treatment with a PD-1 inhibitor was continued after the initiation of administration of an anti-MICA / B antibody.

[0218] In some embodiments, treatment with anti-MICA / B antibodies results in a response (e.g., complete or partial response) or disease stabilization. In some embodiments, the response after administration of anti-MICA / B antibodies lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, or 60 months, or longer.

[0219] In some embodiments, the PD-1 inhibitor includes pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

[0220] In some embodiments, subjects who have previously been treated with PD-1 inhibitors may also have previously been treated with one or more of the following: VEGF inhibitors, EGFR inhibitors, and / or hormone therapy.

[0221] In some embodiments, the subjects being treated are those who have previously been treated with a VEGF inhibitor. In some embodiments, the subjects responded to the VEGF inhibitor (e.g., showed a complete or partial response). In some embodiments, the subjects had a response period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer, after administration of the VEGF inhibitor. In some embodiments, the subjects progressed after the response period to the VEGF inhibitor. In some embodiments, the subjects received treatment with an anti-MICA / B antibody after progression following the response period to the VEGF inhibitor.

[0222] In some embodiments, subjects showed disease stabilization upon administration of VEGF inhibitors. In some embodiments, subjects had disease stabilization periods of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer. In some embodiments, subjects progressed after the disease stabilization period following treatment with VEGF inhibitors. In some embodiments, subjects received treatment with anti-MICA / B antibodies after disease progression following the disease stabilization period after treatment with VEGF inhibitors.

[0223] In some embodiments, treatment with a VEGF inhibitor was discontinued before the initiation of administration of an anti-MICA / B antibody. In some embodiments, treatment with a VEGF inhibitor was continued after the initiation of administration of an anti-MICA / B antibody.

[0224] In some embodiments, treatment with anti-MICA / B antibodies results in a response (e.g., complete or partial response) or disease stabilization. In some embodiments, the response after administration of anti-MICA / B antibodies lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, or 60 months, or longer.

[0225] In some embodiments, the VEGF inhibitor includes a small molecule tyrosine kinase inhibitor (TKI) (e.g., sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib) or an anti-VEGF antibody (e.g., bevacizumab or ranibizumab).

[0226] In some embodiments, subjects who have previously been treated with VEGF inhibitors may also have previously been treated with one or more of the following: PD-1 inhibitors, EGFR inhibitors, and / or hormone therapy.

[0227] In some embodiments, the subjects being treated are those who have previously been treated with an EGFR inhibitor. In some embodiments, the subjects responded to the EGFR inhibitor (e.g., showed a complete or partial response). In some embodiments, the subjects had a response period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer, after administration of the EGFR inhibitor. In some embodiments, the subjects progressed after the response period to the EGFR inhibitor. In some embodiments, the subjects received treatment with an anti-MICA / B antibody after progression following the response period to the EGFR inhibitor.

[0228] In some embodiments, subjects showed disease stabilization upon administration of EGFR inhibitors. In some embodiments, subjects had disease stabilization periods of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer. In some embodiments, subjects progressed after the disease stabilization period following treatment with EGFR inhibitors. In some embodiments, subjects received treatment with anti-MICA / B antibodies after disease progression following the disease stabilization period after treatment with EGFR inhibitors.

[0229] In some embodiments, treatment with an EGFR inhibitor was discontinued before the initiation of administration of an anti-MICA / B antibody. In some embodiments, treatment with an EGFR inhibitor was continued after the initiation of administration of an anti-MICA / B antibody.

[0230] In some embodiments, treatment with anti-MICA / B antibodies results in a response (e.g., complete or partial response) or disease stabilization. In some embodiments, the response after administration of anti-MICA / B antibodies lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, or 60 months, or longer.

[0231] In some embodiments, the EGFR inhibitor includes a small molecule tyrosine kinase inhibitor (TKI) (e.g., gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib) or an anti-EGFR antibody (e.g., cetuximab, panitumumab, or nesitumumab).

[0232] In some embodiments, subjects who have previously been treated with EGFR inhibitors may also have previously been treated with one or more of the following: PD-1 inhibitors, VEGF inhibitors, and / or hormone therapy.

[0233] In some embodiments, the subjects being treated are those who have previously been treated with hormone therapy. In some embodiments, the subjects responded to hormone therapy (e.g., showed a complete or partial response). In some embodiments, the subjects had a response period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer, after administration of hormone therapy. In some embodiments, the subjects progressed after the response period to hormone therapy. In some embodiments, the subjects received treatment with an anti-MICA / B antibody after progression following the response period to hormone therapy.

[0234] In some embodiments, subjects showed disease stabilization upon receiving hormone therapy. In some embodiments, subjects had disease stabilization periods of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer. In some embodiments, subjects progressed after the disease stabilization period following treatment with hormone therapy. In some embodiments, subjects received treatment with anti-MICA / B antibodies after disease progression following the disease stabilization period after treatment with hormone therapy.

[0235] In some embodiments, hormonal therapy was discontinued before the initiation of anti-MICA / B antibody administration. In some embodiments, hormonal therapy was continued after the initiation of anti-MICA / B antibody administration.

[0236] In some embodiments, treatment with anti-MICA / B antibodies results in a response (e.g., complete or partial response) or disease stabilization. In some embodiments, the response after administration of anti-MICA / B antibodies lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, or 60 months, or longer.

[0237] In some embodiments, hormone therapy includes aromatase inhibitors (AIs) (e.g., anastrozole), selective estrogen receptor modulators (SERMs), luteinizing hormone-releasing hormone (LHRH) agonists, antiandrogens, CYP17 inhibitors, progestins, anti-adrenergic drugs, or estrogen receptor antagonists.

[0238] In some embodiments, subjects who have previously been treated with hormone therapy have also previously been treated with one or more of the following: PD-1 inhibitors, VEGF inhibitors, and / or EGFR inhibitors.

[0239] In some embodiments, the subject being treated is a subject with endometrial cancer. In some embodiments, the subject has previously been treated with a PD-1 inhibitor (e.g., an anti-PD-1 antibody). In some embodiments, the subject has previously been treated with a VEGF inhibitor or hormone therapy. In some embodiments, the subject has not previously been treated with a PD-1 inhibitor (e.g., an anti-PD-1 antibody). In some embodiments, the endometrial cancer is HER2+. In some embodiments, the endometrial cancer is ER+ or PR+, or both. In some embodiments, the cancer is a variant of one or more of EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0240] In some embodiments, the subject being treated is a subject having parotid gland cancer. In some embodiments, the parotid gland cancer overexpresses one or both of EGFR and HER2. In some embodiments, the parotid gland cancer is mucoepidermoid carcinoma of the parotid gland. In some embodiments, the cancer is one or more variants of EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0241] cancer

[0242] In certain embodiments, cancers treated according to the methods described herein include, but are not limited to, tumors of the female genitalia, salivary gland tumors, breast cancer, prostate cancer, lung cancer, colon cancer, sarcoma, melanoma, rectal cancer, thyroid cancer, kidney cancer, gastrointestinal cancer, peritoneal cancer, pancreatic cancer, or metastatic lesions thereof. In some embodiments, the tumor is an advanced solid tumor. In some embodiments, the tumor is a head and neck tumor, such as a parotid gland tumor.

[0243] In some embodiments, compositions comprising, but not limited to, an anti-MICA / B antibody are particularly useful for treating high (multiple) tumor volume. Tumor volume refers to the number of cancer cells in the body, the size of the tumor, or the amount of cancer. High tumor volume can be used as a clinical biomarker with negative prognostic assessment in some solid tumors.

[0244] In some embodiments, pharmaceutical compositions containing anti-MICA / B antibodies are particularly useful for treating tumors of the female genitalia. In some embodiments, tumors of the female genitalia are endometrial tumors, ovarian cancer, or cervical cancer. In some embodiments, salivary gland tumors are mucoepidermoid tumors or adenoid cystyosarcoma, such as parotid gland tumors. In some embodiments, cancer is a virus-induced cancer. Virus-induced cancers include those associated with infection with human papillomavirus (HPV), Epstein-Barr virus (EBV), or cytomegalovirus (CMV). In some embodiments, sarcoma is mediastinal endometrial sarcoma or leiomyosarcoma. In some embodiments, peritoneal cancer is peritoneal mesothelioma. In some embodiments, rectal cancer is squamous cell carcinoma of the rectum. In some embodiments, lung cancer is non-small cell lung cancer (NSCLC) or mesothelioma. In some embodiments, gastrointestinal cancer is duodenal cancer. In some embodiments, colorectal cancer is cecal cancer.

[0245] In some embodiments, thyroid cancer is thyroid carcinoma. In some embodiments, cancer is urothelial carcinoma of the bladder. In some embodiments, cancer is adrenocortical carcinoma. In some embodiments, cancer is squamous cell carcinoma of the head and neck. In some embodiments, cervical cancer is squamous cell carcinoma of the cervix or internal cervical cancer. In some embodiments, cancer is esophageal cancer. In some embodiments, cancer is squamous cell carcinoma of the lung. In some embodiments, cancer is a lymphoid neoplasm, diffuse large B-cell lymphoma. In some embodiments, cancer is thymoma. In some embodiments, cancer is acute myeloid leukemia (AML). In some embodiments, cancer is uveal melanoma. In some embodiments, cancer is prostate adenocarcinoma. In some embodiments, cancer is chromophobe renal cell carcinoma. In some embodiments, cancer is low-grade brain glioma. In some embodiments, cancer is hepatocellular carcinoma of the liver. In some embodiments, cancer is skin cutaneous melanoma. In some embodiments, cancer is pheochromocytoma or paraganglioma. In some embodiments, the cancer is uterine carcinosarcoma. In some embodiments, the cancer is ovarian serous cystadenoma. In some embodiments, the cancer is kidney renal papillary carcinoma. In some embodiments, the cancer is glioblastoma multiforme. In some embodiments, the cancer is cholangiocarcinoma. In some embodiments, the cancer is testicular germ cell tumor. In some embodiments, the cancer is invasive breast cancer. In some embodiments, the cancer is kidney renal clear cell carcinoma. In some embodiments, the cancer is lung adenocarcinoma. In some embodiments, the cancer is gastric adenocarcinoma. In some embodiments, the cancer is pancreatic adenocarcinoma. In some embodiments, the cancer is endometrial cancer. In some embodiments, the cancer is rectal adenocarcinoma. In some embodiments, the cancer is colon adenocarcinoma.

[0246] In some embodiments, the cancer is metastatic. In some embodiments, the cancer is one or more variants of EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

[0247] In some embodiments, the cancer is characterized by low levels of PD-L1 expression. In some embodiments, the cancer is characterized by high levels of PD-L1 expression. In some embodiments, the cancer is characterized by low tumor mutational load (TMB). In some embodiments, the cancer is characterized by high tumor mutational load (TMB). In some embodiments, the cancer is immunologically cold. In some embodiments, the cancer is immunologically hot. In some embodiments, the cancer is hormone-sensitive. In some embodiments, the cancer is characterized by overexpression of oncogenic drivers. In some embodiments, the cancer expresses one or more of EGFR, ER, PR, or HER2.

[0248] Dosage and administration regimen

[0249] The therapeutically effective dose or dosage is approximately 0.01 mg / kg to approximately 20 mg / kg, for example, approximately 0.01 mg / kg, approximately 0.02 mg / kg, approximately 0.03 mg / kg, approximately 0.04 mg / kg, approximately 0.05 mg / kg, approximately 0.06 mg / kg, approximately 0.07 mg / kg, approximately 0.08 mg / kg, approximately 0.09 mg / kg, approximately 0.1 mg / kg, approximately 0.2 mg / kg, approximately 0.3 mg / kg, approximately 0.4 mg / kg, approximately 0.5 mg / kg, approximately 0.6 mg / kg, approximately 0.7 mg / kg, approximately 0.8 mg / kg, approximately 0.9 mg / kg, approximately 1.0 mg / kg, approximately 1.1 mg / kg, approximately 1 .2mg / kg, about 1.3mg / kg, about 1.4mg / kg, about 1.5mg / kg, about 1.6mg / kg, about 1.7mg / kg, about 1.8mg / kg, about 1.9mg / kg, about 2mg / kg, about 2.1mg / kg, about 2.2mg / kg, about 2.3mg / kg, about 2.4mg / k g, about 2.5mg / kg, about 2.6mg / kg, about 2.7mg / kg, about 2.8mg / kg, about 2.9mg / kg, about 3mg / kg, about 3.1mg / kg, about 3.2mg / kg, about 3.3mg / kg, about 3.4mg / kg, about 3.5mg / kg, about 3.6mg / kg, about 3.7m g / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4 mg / kg, about 4.1 mg / kg, about 4.2 mg / kg, about 4.3 mg / kg, about 4.4 mg / kg, about 4.5 mg / kg, about 4.6 mg / kg, about 4.7 mg / kg, about 4.8 mg / kg, about 4.9 mg / kg, about 5mg / kg, about 5.1mg / kg, about 5.2mg / kg, about 5.3mg / kg, about 5.4mg / kg, about 5.5mg / kg, about 5.6mg / kg, about 5.7mg / kg, about 5.8mg / kg, about 5.9mg / kg, about 6mg / kg, about 6.1mg / kg, about 6.2mg / kg, Approximately 6.3mg / kg, approximately 6.4mg / kg, approximately 6.5mg / kg, approximately 6.6mg / kg, approximately 6.7mg / kg, approximately 6.8mg / kg, approximately 6.9mg / kg, approximately 7mg / kg, approximately 7.1mg / kg, approximately 7.2mg / kg, approximately 7.3mg / kg, approximately 7.4mg / kg, approximately 7.5mg / kg, about 7.6mg / kg, about 7.7mg / kg, about 7.8mg / kg, about 7.9mg / kg, about 8mg / kg, about 8.1mg / kg, about 8.2mg / kg, about 8.3mg / kg, about 8.4mg / kg, about 8.5mg / kg, about 8.6mg / kg, about 8.7mg / kg, about 8.8 mg / kg, approximately 8.9 mg / kg, approximately 9 mg / kg, approximately 9.1 mg / kg, approximately 9.2 mg / kg, approximately 9.3 mg / kg, approximately 9.4 mg / kg, approximately 9.5 mg / kg, approximately 9.6 mg / kg, approximately 9.7 mg / kg, approximately 9.8 mg / kg, approximately 9.9 mg / kg, approximately 10 mg / kg, approximately 10.1 mg / kg, approximately 10.2 mg / kg, approximately 10.3 mg / kg, approximately 10.4 mg / kg, approximately 10.5 mg / kg, approximately 10.6 mg / kg, approximately 10.7 mg / kg, approximately 10.8 mg / kg, approximately 10.9 mg / kg, approximately 11 mg / kg, approximately 11.1 mg / kg, approximately 11.2 mg / kg mg / kg, approximately 11.3 mg / kg, approximately 11.4 mg / kg, approximately 11.5 mg / kg, approximately 11.6 mg / kg, approximately 11.7 mg / kg, approximately 11.8 mg / kg, approximately 11.9 mg / kg, approximately 12 mg / kg, approximately 12.1 mg / kg, approximately 12.2 mg / kg, approximately 12.3 mg / kg, approximately 12.4 mg / kg, approximately 12.5 mg / kg, approximately 12.6 mg / kg, approximately 12.7 mg / kg, approximately 12.8 mg / kg, approximately 12.9 mg / kg, approximately 13 mg / kg, approximately 13.1 mg / kg, approximately 13.2 mg / kg, approximately 13.3 mg / kg, approximately 13.4 mg / kg, approximately 13. 5 mg / kg, approximately 13.6 mg / kg, approximately 13.7 mg / kg, approximately 13.8 mg / kg, approximately 13.9 mg / kg, approximately 14 mg / kg, approximately 14.1 mg / kg, approximately 14.2 mg / kg, approximately 14.3 mg / kg, approximately 14.4 mg / kg, approximately 14.5 mg / kg, approximately 14.6 mg / kg, approximately 14.7 mg / kg, approximately 14.8 mg / kg, approximately 14.9 mg / kg, approximately 15 mg / kg, approximately 15.1 mg / kg, approximately 15.2 mg / kg, approximately 15.3 mg / kg, approximately 15.4 mg / kg, approximately 15.5 mg / kg, approximately 15.6 mg / kg, approximately 15.7 mg / kg, approximately 15 0.8 mg / kg, approximately 15.9 mg / kg, approximately 16 mg / kg, approximately 16.1 mg / kg, approximately 16.2 mg / kg, approximately 16.3 mg / kg, approximately 16.4 mg / kg, approximately 16.5 mg / kg, approximately 16.6 mg / kg, approximately 16.7 mg / kg, approximately 16.8 mg / kg, approximately 16.9 mg / kg, approximately 17 mg / kg, approximately 17.1 mg / kg, approximately 17.2 mg / kg, approximately 17.3 mg / kg, approximately 17.4 mg / kg, approximately 17.5 mg / kg, approximately 17.6 mg / kg, approximately 17.7 mg / kg, approximately 17.8 mg / kg, approximately 17.9 mg / kg, approximately 18 mg / kg, approximately 18.The intended dosages include 1 mg / kg, approximately 18.2 mg / kg, approximately 18.3 mg / kg, approximately 18.4 mg / kg, approximately 18.5 mg / kg, approximately 18.6 mg / kg, approximately 18.7 mg / kg, approximately 18.8 mg / kg, approximately 18.9 mg / kg, approximately 19 mg / kg, approximately 19.1 mg / kg, approximately 19.2 mg / kg, approximately 19.3 mg / kg, approximately 19.4 mg / kg, approximately 19.5 mg / kg, approximately 19.6 mg / kg, approximately 19.7 mg / kg, approximately 19.8 mg / kg, approximately 19.9 mg / kg, or approximately 20 mg / kg.

[0250] In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 0.1 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg to approximately 10 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 0.1 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 0.3 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 1 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 3 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 6 mg / kg. In some embodiments, the anti-MICA / B antibody is administered at a dose of approximately 10 mg / kg.

[0251] The treatment methods described herein involve one or more administrations of anti-MICA / B antibody in doses disclosed herein. In some embodiments, the method involves one administration of anti-MICA / B antibody. In some embodiments, the method involves two administrations of anti-MICA / B antibody. In some embodiments, the method involves three administrations of anti-MICA / B antibody. In some embodiments, the method involves four administrations of anti-MICA / B antibody. In some embodiments, the method involves five administrations of anti-MICA / B antibody. In some embodiments, the method involves six administrations of anti-MICA / B antibody. In some embodiments, the method involves more than six administrations of anti-MICA / B antibody.

[0252] In some embodiments, the anti-MICA / B antibody is administered according to a dosing interval (e.g., a cycle). In some embodiments, one or more doses of the anti-MICA / B antibody are administered daily. In some embodiments, one or more doses of the anti-MICA / B antibody are administered weekly. In some embodiments, one or more doses of the anti-MICA / B antibody are administered every two weeks (Q2W). In some embodiments, one or more doses of the anti-MICA / B antibody are administered every three weeks (Q3W). In some embodiments, one or more doses of the anti-MICA / B antibody are administered monthly. In some embodiments, one or more doses of the anti-MICA / B antibody are administered every three months. In some embodiments, one or more doses of the anti-MICA / B antibody are administered every six months. In some embodiments, one or more doses of the anti-MICA / B antibody are administered annually.

[0253] In some embodiments, the anti-MICA / B antibody is administered according to a dosing interval (e.g., a cycle). In some embodiments, the dosing interval includes a 3-week cycle, and the anti-MICA / B antibody is administered once every 3 weeks (Q3W). In some embodiments, the dosing interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times. In some embodiments, the repetition of the dosing interval takes place over a period of at least two weeks, three weeks, four weeks, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, twelve months, thirteen months, fourteen months, fifteen months, sixteen months, seventeen months, eighteen months, nineteen months, twenty months, twenty-one months, twenty-two months, twenty-three months, twenty-four months, three years, four years, or five years.

[0254] In some embodiments, the subject is administered one or more drugs as premedication before the administration of the anti-MICA / B antibody.

[0255] In some embodiments, premedication includes acetaminophen or ibuprofen. In some embodiments, subjects are administered acetaminophen or ibuprofen 30 to 60 minutes before administration of the anti-MICA / B antibody. In some embodiments, premedication is administered orally.

[0256] In some embodiments, premedication includes a corticosteroid. In some embodiments, subjects are administered a corticosteroid 30 to 60 minutes before administration of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered before the initial dose of the anti-MICA / B antibody, but not before one or more subsequent doses of the anti-MICA / B antibody. In some embodiments, the corticosteroid is administered orally or intravenously. In some embodiments, the corticosteroid is administered in doses of approximately 2 mg to approximately 50 mg, or in practice, any standard therapeutic corticosteroid dose used in cancer therapy. In some embodiments, the corticosteroid includes dexamethasone. As shown in Example 1 and Figure 5 of this specification, an increase in patient cytokine levels was observed after the initial dose of the anti-MICA / B antibody, but subsequent doses of the anti-MICA / B antibody resulted in a smaller increase or no increase. While we do not wish to be bound by theory, administering corticosteroids prior to the first dose of anti-MICA / B antibodies may weaken the patient's immune response to the antibody administration.

[0257] Any preferred route of administration for use in the manner disclosed herein is intended. In some embodiments, the antibody is administered intravenously. In some embodiments, the antibody is administered subcutaneously. In some embodiments, the antibody is administered topically. In some embodiments, the antibody is administered systemically (e.g., intravenously, intramuscularly, subcutaneously, intradermally, or orally, intranasally, or sublingually). In some embodiments, the antibody is formulated as an ointment, lotion, or emulsion. In some embodiments, the antibody is formulated as a liquid. In some embodiments, the antibody is formulated for topical, oral, buccal, or nasal administration.

[0258] In some embodiments, the anti-MICA / B antibody is administered intravenously over approximately one hour.

[0259] In some embodiments, the individual is monitored before administration of the antibody. Symptoms are identified and their severity is assessed. The antibodies described herein may be administered as a single dose or multiple times over time, alone or in combination with additional treatments, as discussed herein or known to those skilled in the art. In some embodiments, the individual is monitored to determine the effectiveness of the treatment regimen. In some embodiments, the treatment regimen is modified in response to the preliminary outcome of the treatment, thereby changing the dose or frequency of the treatment or the dose and frequency to achieve a desired level of target response, considering symptom reduction, reduction of side effects, or a combination of symptom reduction and reduction of side effects.

[0260] In some embodiments, the treatment methods disclosed herein are monotherapy. In some embodiments, the treatment methods disclosed herein are combination therapy. In some embodiments, combination therapy includes the administration of an anti-MICA / B antibody in combination with another therapeutic agent. In this regard, the ability to completely kill NK cells in a clinical setting to address immunosuppressive TMEs may require the simultaneous involvement of two or more activating stimuli, e.g., CD16A and NKG2D. The ultimate therapeutic effect of NK cell activating therapies in clinical settings may be achieved when delivered in combination with therapies that enhance or supplement NK cell activation, e.g., lenalidomide, cytokines, or checkpoint inhibitors. Combination therapy with checkpoint inhibitors may provide an opportunity for dual activation of both innate and adaptive immune responses, and may reduce exhaustion of both PD-1-expressing T cells and NK cells. Accordingly, this disclosure envisions an immuno-oncological therapy in which anti-MICA / B antibodies may be administered in combination with certain inhibitors of the immune checkpoint molecules: programmed cell death 1 receptor (PD-1), T cell immune receptor with Ig and ITIM domains (TIGIT), T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3), differentiation antigen group 96 (CD96), sialic acid-binding Ig-like lectin 7 (siglec-7), leukocyte-associated immunoglobulin-like receptor 1 (LAIR-1), and inhibitor receptor protein (Irp60). PD-1, considered a typical immune checkpoint molecule, negatively regulates T cell function when it interacts with programmed cell death ligands 1 and 2 (PD-L1 and PD-L2) of immune cells in cancer cells and the tumor microenvironment (TME). This axis has been clinically targeted with antagonist mAb therapy, which has been highly successful. Checkpoint inhibitors may be administered before, after, or concurrently with the anti-MICA / B therapy of this disclosure. Tumor-experienced NK cells have also been shown to express PD-1 and PD-L1 in both preclinical and clinical settings, and thus the use of PD-1 inhibition in combination with MICA / B antibodies constitutes a particular embodiment intended herein.

[0261] In certain embodiments, the compositions of the present disclosure that include anti-MICA / B antibodies are in a combination pharmaceutical or drug therapy with at least one additional therapeutic agent selected from the group consisting of antibodies, antibody fragments, antibody conjugates, cytotoxic agents, toxins, radionuclides, immunomodulatory molecules, photoactive therapeutic agents, radiosensitizers, hormones, anti-angiogenic agents, and combinations thereof.

[0262] In some embodiments, the therapeutic agent includes a chemotherapeutic agent. In some embodiments, chemotherapeutic agents include, but are not limited to, cytotoxic agents, antimetabolites (e.g., folic acid antagonists, purine analogs, pyrimidine analogs, etc.), topoisomerase inhibitors (e.g., camptothecin derivatives, anthracenediones, anthracyclines, epipodophyllotoxins, quinoline alkaloids, etc.), microtubule inhibitors (e.g., taxanes, vinca alkaloids), protein synthesis inhibitors (e.g., cephalotaxine, camptothecin derivatives, quinoline alkaloids), alkylating agents (e.g., alkyl sulfonates, ethyleneimines, nitrogen mustards, nitrosoureas, platinum derivatives, triazenes, etc.), alkaloids, terpenoids, kinase inhibitors, and immune checkpoint inhibitors.

[0263] In some embodiments, the anti-MICA / B antibodies disclosed herein are administered in combination with a therapeutic agent that binds to CD16A. Those skilled in the art recognize many chimeric, human, and humanized IgG1 antibody therapeutics that bind to CD16A, including, but not limited to, agents such as rituximab (targeting CD20), daratumumab (CD38), and trastuzumab (human epidermal growth factor receptor 2 - HER2).

[0264] In some embodiments, the anti-MICA / B antibodies disclosed herein are administered in combination with a therapeutic agent that induces an immune response. In some embodiments, the anti-MICA / B antibodies disclosed herein are administered in combination with a therapeutic agent that inhibits the downregulation of the immune response. In some embodiments, inducing an immune response includes activation or upregulation of NK cell activity. In some embodiments, inducing an immune response includes activation or upregulation of T cell activity. In some embodiments, the immune checkpoint inhibitor target includes PD-1. In some embodiments, the immune checkpoint inhibitor target includes PD-L1.

[0265] Non-specific examples of immune checkpoint inhibitors targeting the PD-1 / PD-L1 pathway include, but are not limited to, pimivalimab, pembrolizumab (Keytruda, MK-3475, lambrolizumab), nivolumab (e.g., Opdivo, BMS-936558, MDX1106), semiprimab, spartalizumab (PDR001), STI-A1110, AMP-224, AMP-514 (MEDI0680), JS00 Examples include atezolizumab (e.g., Tecentriq, MPDL3280A), avelumab (MSB0010718C), durvalumab ((MEDI4736), BMS-936559), pizilizumab (CT-011), BMS-936559 (MDX1105), and LY3300054, and CK-301. In some embodiments, the PD-1 inhibitor includes pembrolizumab. In other embodiments, the PD-1 inhibitor includes nivolumab.

[0266] In some embodiments, pembrolizumab is QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREP It comprises a heavy chain containing the amino acid sequence QVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 11), and a light chain containing the amino acid sequence EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12).

[0267] In the following discussion, pembrolizumab is used as an exemplary PD-1 inhibitor. It should be understood that alternative PD-1 inhibitors, or in fact PD-L1 inhibitors, may be used instead of pembrolizumab. In some embodiments, pembrolizumab is administered intravenously to the subject at a dose of approximately 200 mg. In some embodiments, pembrolizumab is administered to the subject once every three weeks (Q3W). In some embodiments, the subjects receiving combination therapy including an immune checkpoint inhibitor (e.g., pembrolizumab) have ovarian cancer, prostate cancer, colorectal cancer, NSCLC, squamous cell carcinoma, or cervical cancer. In some embodiments, the anti-MICA / B antibody described herein is administered in combination with a VEGF inhibitor. In some embodiments, the VEGF inhibitor includes a small molecular weight tyrosine kinase inhibitor (TKI) (e.g., sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib) or an anti-VEGF antibody (e.g., bevacizumab or ranibizumab). In some embodiments, the anti-MICA / B antibody described herein is administered in combination with an EGFR inhibitor. In some embodiments, the EGFR inhibitor includes a small molecular weight tyrosine kinase inhibitor (TKI) (e.g., gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib) or an anti-EGFR antibody (e.g., cetuximab, panitumumab, or nesitumumab). In some embodiments, the anti-MICA / B antibody described herein is administered in combination with hormone therapy. In some embodiments, hormone therapy includes aromatase inhibitors (AIs) (e.g., anastrozole), selective estrogen receptor modulators (SERMs), luteinizing hormone-releasing hormone (LHRH) agonists, antiandrogens, CYP17 inhibitors, progestins, anti-adrenergic agents, or estrogen receptor antagonists. In certain embodiments, the anti-MICA / B antibodies described herein are administered in combination with radiotherapy. In certain embodiments, the anti-MICA / B antibodies described herein are administered in combination with one or more cell therapies.

[0268] Table 2 shows exemplary chemical structures of certain therapeutic agents described herein.

[0269] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0270] Pharmaceutical composition

[0271] Pharmaceutical compositions comprising an anti-MICA / B antibody disclosed herein and a pharmaceutically acceptable carrier or excipient are also disclosed herein.

[0272] In some embodiments, excipients for use with the compositions disclosed herein include maleic acid, tartaric acid, lactic acid, citric acid, acetic acid, sodium bicarbonate, sodium phosphate, histidine, glycine, sodium chloride, potassium chloride, calcium chloride, zinc chloride, water, dextrose, N-methylpyrrolidone, dimethyl sulfoxide, N,N-dimethylacetamide, ethanol, propylene glycol, polyethylene glycol, diethylene glycol monoethyl ether, and the surfactant polyoxyethylene sorbitan monooleate.

[0273] In some embodiments, the composition further comprises additional therapeutic agents. In some embodiments, the therapeutic agent is a chemotherapeutic agent. Examples of chemotherapeutic agents include, among others, cytotoxic agents, antimetabolites (e.g., folate antagonists, purine analogs, pyrimidine analogs, etc.), topoisomerase inhibitors (e.g., camptothecin derivatives, anthracendions, anthracyclines, epipodophyllotoxins, quinoline alkaloids, etc.), microtubule inhibitors (e.g., taxanes, vinca alkaloids), protein synthesis inhibitors (e.g., cephalotaxine, camptothecin derivatives, quinoline alkaloids), alkylating agents (e.g., alkyl sulfonates, ethyleneimines, nitrogen mustard, nitrosourea, platinum derivatives, triazenes, etc.), alkaloids, terpenoids, and kinase inhibitors.

[0274] In some embodiments, the antibody and the therapeutic agent are present in the same formulation. In some embodiments, the antibody and the therapeutic agent are present in different formulations. In some embodiments, the antibody described herein is used before the administration of other therapeutic agents. In some embodiments, the antibody described herein is used simultaneously with the administration of other therapeutic agents. In some embodiments, the antibody described herein is used after the administration of other therapeutic agents.

[0275] In some embodiments, pharmaceutical formulations are prepared to be adapted to specific local, regional, or systemic administration or delivery routes. Therefore, pharmaceutical formulations include carriers, diluents, or excipients suitable for administration via specific routes. Non-limiting specific examples of administration routes for compositions herein include parenteral, e.g., intravenous, intra-arterial, intradermal, intramuscular, subcutaneous, intrapleural, transdermal (topical), transmucosal, intracranial, intraspinal, intraocular, rectal, oral (gastrointestinal), mucosal administration, and any other formulations suitable for treatment methods or administration protocols.

[0276] In some embodiments, solutions or suspensions used for parenteral administration include sterile diluents such as water for injection, saline solution, fixative oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfate; chelating agents such as ethylenediaminetetraacetic acid; buffering agents such as acetates, citrates, or phosphates; and agents for adjusting tonicity such as sodium chloride or dextrose. In some embodiments, pH is adjusted using an acid or base, such as hydrochloric acid or sodium hydroxide.

[0277] The injectable pharmaceutical formulation comprises a sterile aqueous solution (if water-soluble) or dispersion and a sterile powder for immediate preparation of a sterile injection solution or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL® (BASF, Parsippany, NJ), or phosphate-buffered saline (PBS). In some embodiments, the carrier is a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), or suitable mixtures thereof. Fluidity is maintained in some embodiments, for example, by the use of a coating agent such as lecithin, by maintaining the required particle size in the case of a dispersion, and by the use of a surfactant. Examples of antibacterial and antifungal agents include parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal. Isotonic agents, such as sugars, polyhydric alcohols such as mannitol or sorbitol, or sodium chloride, are included in some embodiments of the composition. In some cases, the formulation may include agents that delay absorption; in some embodiments, for example, aluminum monostearate or gelatin prolongs the absorption of the injectable composition.

[0278] In some embodiments, the sterile injectable preparation is prepared by incorporating the required amount of the active composition into a suitable solvent having one or a combination of the above components. Generally, the dispersion is prepared by incorporating the active composition into a sterile vehicle containing a basic dispersion medium and any other components. In the case of sterile powders for the preparation of sterile injection solutions, methods of preparation include, for example, vacuum drying and lyophilization to obtain the powders of the active ingredient and any additional desired components from their previously prepared solutions.

[0279] For transmucosal or transdermal administration, suitable penetrants for the barrier to be permeated are used in the formulation. Such penetrants are known in the art and include, for example, surfactants, bile salts, and fusidic acid derivatives for transmucosal administration. In some embodiments, transmucosal administration is achieved via the use of nasal sprays, inhalation devices (e.g., aspirators) or suppositories. For transdermal administration, the active compound is formulated into ointments, plasters, gels, creams or patches.

[0280] In some embodiments, the pharmaceutical preparation is prepared with carriers that prevent rapid elimination from the body, such as controlled release formulations or time delay substances such as glyceryl monostearate or glyceryl stearate. The preparation may also, in some embodiments, be delivered using manufactured articles such as implants and microencapsulation delivery systems for local, regional or systemic delivery, or to achieve controlled or sustained release.

[0281] Since there remains a significant unmet need for patients with NSCLC having targetable oncogenic mutations that recur after TKI, the inventors recognize potential benefits in novel therapies that can be readily combined with established CPI. The combination of CLN-619 and pembrolizumab involves multiple immune effector cells, including natural cells by CLN-619 and T cells by pembrolizumab. The safety profile of CLN-619 with a biological rationale for combination with CPI makes this a potentially synergistic approach.

[0282] Initial clinical findings suggest that the combination of CLN-619, a novel antibody targeting MICA / B, and pembrolizumab may benefit patients whose cancer is typically unsuitable for checkpoint inhibitor therapy. More specifically, we observed objective responses in patients with ALK and EGFR-mutated NSCLC that relapsed after tyrosine kinase inhibitor (TKI) therapy, and in patients who typically do not respond to checkpoint inhibitors.

[0283] In addition, longer-term follow-up of patients treated with CLN-619 monotherapy demonstrates favorable safety and sustained clinical utility through extended treatment, including objective responses and long-term disease stabilization in patients with multiple tumor types and disease progression after CPI therapy.

[0284] [Examples]

[0285] The following embodiments are provided for the purpose of illustrating various embodiments of the present invention and are not intended to limit the invention in any way. Herein, the embodiments involving the methods described herein are representative and illustrative of a particular embodiment and are not intended to limit the scope of the invention. Modifications and other uses of embodiments that fall within the scope of the spirit of the invention as defined by the claims will be conceivable to those skilled in the art.

[0286] Example 1. CLN-619 (anti-MICA / B antibody) promotes innate immune cell-mediated antitumor activity.

[0287] CLN-619 is a humanized IgG1 monoclonal antibody that targets MICA and MICB (MICA / B) and is currently in Phase 1 clinical development in cancer patients (NCT05117476; see the examples below). These examples demonstrate that this antibody promotes the antitumor activity of innate immune cells.

[0288] MICA / B functions as an activation signal on target cells for recognition by the NKG2D receptor, which is expressed in subsets of NK and T cell populations. MICA / B expression is induced in response to stress, thereby enabling NKG2D-mediated elimination of target cells. In NK cells, NKG2D is one of many receptors in a complex network of activating and inhibitory receptors, so that NKG2D pathway activation leads to cytokine production, enhancement of ADCC, and target cell death. In CD8 T cells, the NKG2D axis plays a co-stimulatory role in lowering the lysis threshold when TCR is involved, and can also drive direct CD8-mediated death after preceding TCR activation. MICA / B is widely expressed in a variety of solid and hematological malignancies. However, tumor cells evade NKG2D-mediated elimination by shedding the MICA / B ligand from their cell surface via proteases present in the tumor microenvironment (TME). CLN-619 functions by binding to MICA / B and preventing shedding, thereby increasing MICA / B cell surface expression, restoring the NKG2D-MICA / B axis, and promoting tumor cell death by NK cells and T cells. In addition, CLN-619 can mediate ADCC and ADCP by NK cells and macrophages. In a preclinical xenograft model, tumor growth was inhibited at a low dose of 0.03 mg / kg.

[0289] The inventors investigated the effects of CLN-619 on various primary immune cells, including NK cells, T cells, and macrophages. In the evaluation of NK cells, CLN-619 regulation of cytokine production and cytotoxicity was measured in the presence of MICA / B-expressing target cells. In addition, this example demonstrates the contribution of CLN-619's Fc-mediated function. In the context of T cells, the requirements for TCR co-stimulation or prior antigen exposure in the activation of the NKG2D pathway by CLN-619 were explored. In the context of macrophages, the inventors measured CLN-619's ability to mediate ADCP to MICA / B-expressing target cells. These data highlight the potential of CLN-619 to bind to multiple immune cell types within the TME, which may result in greater and broader efficacy compared to IO therapies targeting a single immune cell type.

[0290] A. Method

[0291] CLN-619 was characterized for its binding epitope and affinity, its effects on MICA / B surface and solubility levels, and its in vitro tumor cell death properties. In a mouse model, this mAb was tested for tumor growth inhibition. The contribution of the functional Fc-gamma 1 (Fcγ1) domain to CLN-619 activity was also evaluated.

[0292] Generation of CLN-619

[0293] To create a diverse panel of monoclonal antibodies against human MICA / B protein, a repetitive immunizations multiple sites (RIMMS) immunization protocol was performed in Swiss James Lambert (SJL) mice. The full-length extracellular (ECD) of the most common allele variant (MICA*008) was used as the antigen [Klussmeier A, et al., Front Immunol. 2020;11]. Lead candidates were humanized using in silico modeling.

[0294] Octet binding evaluation of CLN-619 to MICA and MICB

[0295] To determine monovalent affinity, CLN-619 antibody (10 μg / ml) was captured in a Dip and Read α anti-human IgG Fc capture biosensor. The sensor was then incubated with purified MICA*001, MICA*002, MICA*004, MICA*008, or MICB*004 ECD proteins. Ka, kd, and KD were calculated using a fitting algorithm (ForteBio analysis suite 8.0) assuming 1:1 binding. MICA / B protein similarity was determined using the BLOSUM62 algorithm.

[0296] ELISA evaluation of CLN-619 binding to MICA / B allele variants

[0297] 96-well plates were coated with monomer his-tagged MICA / B as a capture antigen, followed by the addition of serial dilutions of CLN-619. Unbound material was washed, and horseradish peroxidase (HRP) conjugated mouse anti-human IgG Fc was added to the wells for detection.

[0298] Luminex evaluation of CLN-619 binding to MICA

[0299] The binding of CLN-619 (10 ug / mL) and the positive control antibody 6D4 to 28 different allele variants of MICA was evaluated using Luminex (LifeCodes LSA MIC Luminex kit) [Ghadially H, et al. Br J Cancer. 2017;116:1208-17]. 6D4 was detected using a goat anti-mouse secondary antibody. Background MFI was subtracted from raw MFI, and the two sets of samples were averaged.

[0300] X-ray crystal structure analysis of MICA-CLN-619

[0301] X-ray diffraction data were collected from Fab CLN-619:MICA crystals using optimized low-temperature conditions at SWISS LIGHT SOURCE (SLS, Villigen, Switzerland). The structure of the complex of human Fab antibody fragment CLN-619 and the MICA alpha-3 domain (Fab CLN-619:MIC-A) was determined at a resolution of 2.12 Å.

[0302] Flow cytometry evaluation of surface MICA / B levels after CLN-619 treatment

[0303] HCC1534, PLC / PRF / 5, and HCT-116 cells were incubated for 24 hours with serially diluted CLN-619, CLN-619 DANA, or hIgG1 control antibodies. After washing, cells were stained with live / dead color-determination dyes. Cell surface MICA / B was detected using 6D4 antibody, an anti-human MICA / B-PE antibody that is non-competitive with CLN-619. Mouse IgG2a-PE isotype antibody was used as a control. Surface MICA / B levels were quantified using Quantibrite beads. Dose-response curves were fitted using a 4-parameter logistic regression model.

[0304] Evaluation of soluble MICA / B levels after CLN-619 treatment

[0305] HCC1534, PLC / PRF / 5, and HCT-116 were plated and treated in two denominations for 24 hours with serially diluted CLN-619 or hIgG1. The supernatant was collected for analysis of sMICA levels in enzyme-linked immunosorbent assay (ELISA). The antibodies used in ELISA were non-competitive with CLN-619. The ELISA plates were coated with BAMO1 capture antibody. A standard curve and supernatant sample of recombinant human MICA*001 ECD were added to the plates. Biotinylation detection antibody 10E9.H6 was added to the plates, followed by streptavidin-horseradish peroxidase (HRP). EC50 values ​​were calculated using a 4-parameter nonlinear fit model in GraphPad Prism.

[0306] Primary NK cell death assay

[0307] The ability of CLN-619 to mediate NK cell-mediated tumor cell death was evaluated in a co-culture assay of purified human primary NK cells and MICA-expressing HCC1534 tumor cells in the xCELLigence system. NK cells were isolated via negative selection from a frozen PBMC stock derived from a healthy human donor, and their purity was confirmed. HCC1534 target cells were plated onto E-96 Xcelligence glass plates with antibody titrated stepwise. After HCC1534 cells adhered, NK cells were added at an effector-to-target cell ratio (E:T) of 40:1, and HCC1534 cell death was monitored for 72 hours. EC50 values ​​were calculated using a 4-parameter nonlinear fit model in GraphPad Prism.

[0308] ADCC Reporter Assay

[0309] The ADCC activity of CLN-619 was measured using a reporter assay (Promega). Jurkat cells expressing high (V) or low-affinity (F) FcγRIIIa were engineered to express an activated T cell nuclear factor (NFAT)-luciferase reporter construct. HCT-116 or HCC1534 cells were treated with serial dilutions of CLN-619 or IgG1 control antibody. Engineered Jurkat cells were added at a 10:1 E:T ratio for 6 hours. EC50 values ​​were calculated using a 4-parameter nonlinear fit model in GraphPad Prism.

[0310] Primary cell ADCP assay

[0311] CD14+ monocytes were collected from two healthy PBMC donors by magnetic selection using RoboSep (Protocol 17858) or manual isolation (StemCell). Macrophages were differentiated for 6 days, then harvested and seeded with M-CSF supplemented medium. HCC1534 or HCT-116 eFluor670-labeled target cells and macrophage effector cells were plated in a 0.5:1 E:T ratio and incubated for 2 hours with serially diluted CLN-619 or the highest dose of hIgG1 or cetuximab control antibody. Post-incubation, macrophages were sequentially incubated with viability-determinating dye, blocking buffer (Biolegend), and BV510-labeled anti-CD64 antibody. ADCP was determined by gating target cell fluorescence (eF670+) within CD64+ cells via flow cytometry.

[0312] Flow cytometry evaluation of MICA binding to NKG2D on NK cells

[0313] NK cells were purified from healthy donor PBMCs by negative selection. Serially diluted CLN-619, CLN-619 DANA, or hIgG1 antibody were pre-incubated with his-tagged MICA ECD in a 10:1 ratio. The antibody-MICA conjugates were then added to NK cells in the presence or absence of anti-NKG2D antibody. MICA bound to NK cells was detected by anti-His antibody.

[0314] Biacore evaluation of CLN-619 against Fc gamma receptor

[0315] To evaluate CLN-619 binding to Fc gamma receptors (FcγR), recombinant His6-tagged FcγR was immobilized on an anti-His6 antibody-coated CM5 chip. CLN-619 WT or CLN-619 DANA was passed through the biosensor. ka, kd, and KD were calculated in steady state for all FcR except CD64, and these were evaluated as 1:1 affinity.

[0316] In vivo efficacy study

[0317] BALB / c SCID mice were inoculated with 10 × 10⁶ HCC1534 cells. Mice were treated with hIgG1 (10 mg / kg), CLN-619 (or 0.03, 0.3, 3.0, 10 mg / kg), or CLN-619 DANA (0.03, 0.3, 3.0 mg / kg). Medication was started on the same day as tumor cell transplantation and continued twice weekly for a total of 8 doses. BALB / c mice were subcutaneously inoculated with 5 × 10⁶ PLC / PRF / 5 cells. Mice were administered intracellularly via either vehicle (PBS) or CLN-619 (0.3 mg / kg or 3 mg / kg) three times weekly for a total of 16 doses.

[0318] In both studies, tumor size and body weight were measured twice weekly throughout the study period. The HCC1534 and PLCR / PRF / 5 studies were concluded on days 35 and 39, respectively. Statistical analysis was performed using two-way ANOVA with multiple comparisons (GraphPad).

[0319] Luciferase-tagged HCT-116 MICA / B-expressing human colorectal cancer cells were inoculated via intravenous injection (IP). On day 4, treatment was initiated with IP injection of either hIgG1 or CLN-619 at a dose of 10 mg / kg twice weekly for 5 and 6 weeks, respectively. Bioluminescence imaging was performed twice weekly to monitor the progression of disseminated disease. Survival analysis was performed based on day 40, when the last mouse reached the mortal state. Statistical analysis was performed using the log-rank test.

[0320] Pharmacokinetics

[0321] BALB / c SCID mice were administered 1, 3, or 10 mg / kg of CLN-619 via ip dose, and blood samples were collected by mandibular hemorrhage at the following time points during the study: 1, 3, 6, 24, 48, 72, 96 hours and 11, 14, 21, and 28 days. A total of 3 or 4 mice were evaluated at each time point for each dose group. Serum concentrations were measured by MICA / B sandwich ELISA.

[0322] B. Results

[0323] CLN-619 bound to the alpha-3 domain of MICA / B with high affinity without interfering with its interaction with NKG2D on NK cells. In vitro, CLN-619 increased the level of MICA / B cell surface expression while simultaneously decreasing the level of soluble MICA / B. Treatment of cancer cell lines with CLN-619 induced antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cell-mediated phagocytosis (ADCP), with EC50 values ​​dependent on MICA / B expression levels. CLN-619 resulted in potent inhibition of tumor growth in liver and lung mouse xenograft models and increased mouse survival in a disseminated colorectal cancer model.

[0324] CLN-619 binds to the alpha-3 domain of MICA / B.

[0325] CLN-619 was derived from a parental mouse antibody selected for its high affinity binding to the alpha-3 domain of MICA and its ability to amplify the cell surface expression of MICA / B. The variable domain of the parental antibody was humanized and introduced into the human IgG1 backbone to generate CLN-619.

[0326] The MICA / B gene is highly polymorphic, with nearly 150 MICA alleles and 50 MICB alleles existing in the human population. Standard MICA*001 and MICB*004 alleles are 83% identical in their overall protein sequence and 91% identical in their alpha-3 domains. Given the highly polymorphic nature of the protein, confirming the broad reactivity of CLN-619 was important. The monovalent affinity of CLN-619 to the extracellular domains (ECDs) of several common human MICA alleles (MICA*001, *002, *004, and *008) and MICB allele*004 was measured using Octet. The KD values ​​of CLN-619 binding to recombinant MICA allele variants ranged from 0.77 to 2.04 nM, all within a 3x range (Figure 1A). The KD value of CLN-619 binding to MICB allele*004 was 11.37 nM, which was approximately 1 / 5 to 1 / 15 of that of the MICA allele. ELISA experiments independently confirmed the binding of CLN-619 to representative allele variants and demonstrated similar binding affinities between MICA and MICB alleles (Figure 1B). The broad reactivity of CLN-619 was further confirmed in a Luminex-based assay evaluating 28 recombinant human MICA ECD proteins representing the most common alleles (Figure 1C). CLN-619 showed similar affinity to all alleles except for MICA allele*046, a rare allele present in less than 0.01% of the population, where reduced binding was observed compared to other alleles (Figure 8). Overall, CLN-619 showed broad reactivity across all MICA alleles tested, representing 89% of the population.

[0327] X-ray crystallography was performed to elucidate the interaction between CLN-619 and the alpha-3 domain of MICA at atomic resolution. Cocrystal structures of the CLN-619 Fab fragment and the MICA alpha-3 domain of MICA allele*001 were obtained at a resolution of 2.12 angstroms. CLN-619 recognized a discontinuous epitope in the alpha-3 domain defined as Thr204~Ser297 and interacted with a total of 19 amino acid residues, accounting for approximately 20% of the alpha-3 domain. Notably, some of the known cleavage sites within the alpha-3 domain of MICA / B overlap with the epitopes recognized by CLN-619. The CLN-619 epitope is highly conserved among the most common MICA allele variants, and amino acid changes were observed in only 4 of the 19 interacting amino acid residues, with the majority of these changes being relatively conserved (Figure 10). Importantly, CLN-619 bound to the alpha-3 domain of MICA in a region and angle that did not appear to sterically interfere with the binding of the alpha-1 and alpha-2 domains to the NKG2D dimer (Figure 2). In flow-based assays, CLN-619 did not interfere with the NKG2D-MICA interaction, supporting crystallographic analysis (Figure 11). Interestingly, when MICA formed a complex with CLN-619, increased NK cell binding was observed compared to MICA alone, which likely reflected the co-binding of CLN-619 to both NKG2D and CD16A on NK cells. When an anti-NKG2D antibody was included in the experimental conditions, MICA binding to primary NK cells was significantly reduced.

[0328] CLN-619 amplifies the cell surface expression of MICA / B in tumor cells.

[0329] Based on structural data, the inventors hypothesized that CLN-619 binding to MICA / B at the alpha-3 domain prevents protease access to the MICA / B ligand, thereby suppressing shedding and increasing cell surface expression. The inventors evaluated the ability of CLN-619 to modulate cell surface and soluble MICA / B levels in cell-based assays using human cancer cell lines HCC1534 (lung), PLC / PRF / 5 (liver), and HCT-116 (colorectal). These cancer cell lines express approximately 58,000, 17,000, and 10,000 copies of MICA / B on their cell surfaces, respectively, as measured by flow cytometry. After treatment with CLN-619, cell surface levels of MICA / B were quantified by flow cytometry, and sMICA / B levels in the culture medium were measured by ELISA. Unlike control hIgG1, CLN-619 amplified MICA / B surface expression (Figure 3A) and simultaneously dose-dependently reduced sMICA levels (Figure 3B). The EC50 values ​​for increased MICA / B surface expression were 54, 69, and 51 ng / mL CLN-619 for the HCC1534, PLC / PRF / 5, and HCT-116 cell lines, respectively. The IC50 values ​​for inhibition of sMICA / B release into cell culture medium were 107, 162, and 79 ng / mL CLN-619 for the three cell lines, respectively. Notably, the similar EC50 values ​​for the enhancement of MICA / B on the cell surface by CLN-619, and the IC50 values ​​for the reduction of sMICA / B in the supernatant, support a causal relationship.

[0330] CLN-619 mediates NK cell-mediated cytokine production and target cell lysis.

[0331] The ability of CLN-619 to stabilize MICA / B ligands on the surface of tumor cells suggests that CLN-619 may be able to amplify NK cell activation due to an increase in the level of ligands binding to the activated NKG2D receptor. The effects of CLN-619 on NK cell-mediated cytokine release and target cell lysis were evaluated in a co-culture assay using primary NK cells from two healthy PBMC donors in the presence of HCC1534 target cells. In a co-culture system containing primary NK cells and MICA / B-expressing tumor cells, CLN-619 induced dose-dependent IFNγ release and potent target cell lysis, with EC50 values ​​of 13–19 ng / mL (Figure 4A) and EC50 values ​​of 1–2 ng / mL for target cell lysis (Figure 4B). Activity was dependent on the complete Fc domain.

[0332] We generated an Fc-silencing form of CLN-619, called CLN-619 DANA, and evaluated the contribution of FcγRγ binding to CLN-619-mediated NK cell activation and tumor cell death. CLN-619 DANA contains two mutations in its Fc domain, D265A and N297A, which have been previously shown to eliminate FcγR binding to IgG1 antibodies. While the DANA mutations resulted in the loss of FcγR binding by CLN-619, the ability to bind to MICA and regulate cell surface levels was retained (Figures 9 and 12). When evaluated in a primary NK cell co-culture assay, CLN-619 DANA failed to induce significant IFN release or target cell lysis (Figures 4A and 4B). These results demonstrate the necessity of a complete Fcγ1 domain for CLN-619-mediated immune activation and target cell lysis by NK cells.

[0333] CLN-619 induces ADCC and ADCP in MICA / B-expressing tumor cells.

[0334] Considering that CLN-619 is a human IgG1 antibody possessing a functional Fc domain, the inventors evaluated whether CLN-619 can mediate various Fc-mediated functional activities such as ADCC, ADCP, and CDC. Functional binding of FcγRIIIa / CD16A by CLN-619 was investigated in a cell-based reporter assay designed as a surrogate for evaluating ADCC activity. Jurkat cell lines, which express luciferase genes driven under the control of NFAT transcription factors responsive to human FcγRIIIa / CD16A activation, were used as reporter effector cells. Jurkat cell lines (one expressing the high-affinity V158 variant of CD16A and the other expressing the low-affinity F158 variant) were co-cultured with MICA / B-expressing target cells in the presence of CLN-619. The most potent CD16A signaling was observed in co-culture with HCC1534, a tumor cell line expressing high levels of MICA / B (Figure 5). Upon CLN-619 treatment, EC50 values ​​ranged from 8 to 81 ng / ml, and the signal was consistently lower for the F variant of CD16A compared to the V variant. Consistent with its functionally deficient Fc domain, CLN-619 DANA was inactive in the Jurkat cell assay. Overall, the EC50 values ​​for CLN-619-induced CD16A activation in the reporter cell line correlated with MICA / B expression levels in target cells, suggesting that CLN-619 can target-dependently mediate ADCC.

[0335] Considering CLN-619's ability to bind to FcγRIIA / CD32A, a FcγR receptor primarily responsible for ADCP, we also evaluated CLN-619's ability to induce macrophage-mediated phagocytosis

[20] . Human peripheral macrophages polarized to the M1 phenotype were co-cultured with fluorescently labeled HCC1534 or HCT-116 tumor cells. Phagocytosis in response to CLN-619 was measured by flow cytometry. Since both cell lines were confirmed to express EGFR, the anti-EGFR mAb cetuximab was included as a positive control. Treatment with CLN-619 resulted in dose-dependent phagocytosis of labeled tumor cells, similar to cetuximab, but with minimal phagocytosis observed with isotype control antibodies (Figure 6).

[0336] CLN-619 was able to mediate ADCP and ADCC, but no complement-dependent cytotoxicity (CDC) activity was observed in response to CLN-619 treatment (data not shown). However, CLN-619 treatment resulted in dose-dependent phagocytosis of labeled MICA / B-expressing tumor cells in a co-culture assay with primary macrophages.

[0337] Antitumor activity of CLN-619

[0338] Given the potent antitumor cell-killing activity of CLN-619 observed in vitro, the effect of CLN-619 on tumor growth in vivo was evaluated in immunodeficient BALB / c SCID mice using human tumor xenografts expressing MICA / B. Exposure to CLN-619 after intraperitoneal (ip) administration to BALB / c SCID mice was dose-proportional, and the serum half-life was consistent with the serum half-life of a typical human IgG1 antibody administered to the mice.

[0339] In a PLC / PRF / 5 liver tumor xenograft model, intravenous administration of either 0.3 mg / kg or 3 mg / kg of CLN-619 resulted in complete inhibition of tumor growth compared to the control group (p=0.015) (Figure 7A). In an HCC1534 lung xenograft model, intravenous administration of CLN-619 at dose levels ranging from 0.3 to 10 mg / kg demonstrated statistically significant (p<0.0001) dose-dependent antitumor activity compared to treatment with isotype control mAbs, with tumor growth inhibition rates (TGI) ranging from 45 to 87%. No antitumor activity was observed at any dose level in CLN-619 DANA (Figure 7B). These data suggest that the complete Fcγ1 domain of CLN-619 is essential for its antitumor activity in vivo, which is consistent with our in vitro observations in NK cell co-culture assays.

[0340] The activity of CLN-619 in metastatic situations was explored in a mouse model designed to mimic late-stage disseminated disease. Luciferase-expressing HCT-116 human colorectal cancer cells were inoculated into mice via ip (intracellular matrix) to enable tumor cell dissemination to distal organs. Kaplan-Meier survival analysis demonstrated that CLN-619 treatment provided a statistically significant survival benefit compared to hIgG1 (p<0.0001) (Figure 7C).

[0341] C. Conclusion

[0342] CLN-619 effectively restores cytotoxic signaling pathways in immune cells by inhibiting MICA / B shedding. Potent antitumor activity of CLN-619 as monotherapy was observed in several preclinical models. The activity of CLN-619 requires a functional Fcγ1 domain, suggesting that simultaneous binding of NKG2D and CD16A in immune cells is necessary for optimal cytotoxicity. The preclinical data reported in this example support the evaluation of CLN-619 in cancer patients.

[0343] The data presented herein reveal the diverse mechanisms of action of CLN-619. Apart from inhibition of MICA / B shedding and increased expression of NKG2D ligand in cancer cells, we observed the ADCC and ADCP functionality of CLN-619 that is dependent on the wild-type hIgG1 scaffold. The Fc-deficient morph of CLN-619 showed no activity in the NK co-culture cell death assay despite retention of NKG2D binding (Figure 11) (Figure 4B). CLN-619 DANA also lacked activity in vivo. We hypothesize that the activity of CLN-619 may require dual stimulation of CD16A and NKG2D receptors in NK cells. This dual receptor binding concept is supported by published studies showing that NKG2D binding can synergistically enhance ADCC and amplify CD16A and NKp46 activation in resting NK cells.

[0344] CLN-619 demonstrated attractive monotherapy activity at low doses in multiple tumor xenograft models using tumor cell lines representing indications in which the NKG2D pathway has been shown to be clinically relevant, such as HCC and NSCLC. Human xenograft models were selected because, although mice do not possess an orthologue of MICA / B, mouse NKG2D on NK cells has been shown to recognize MICA / B on human tumor cells. The host mice used in our studies possess qualified NK cells and macrophages but lack T cells. Therefore, the observed antitumor efficacy is most likely attributable to target cell lysis by NK cells and / or phagocytosis by macrophages. Contributions to efficacy by NKG2D-expressing T cell subtypes, such as CD8+ T cells, NKT cells, and γδ T cells, may only occur in fully immunoqualified models. Notably, the NKG2D axis has been shown to play a co-stimulatory role in lowering the threshold for CD8+ T cell lysis, and can even drive CD8+ T cell-mediated lysis of cancer cells after preceding TCR activation in the absence of MHC expression. CLN-619 is likely to be able to promote antitumor activity by binding to various cytotoxic immune effector cells in the TME. In a co-culture system containing CD8+ T cell enrichment populations and MICA / B-expressing tumor cells, CLN-619 induced dose-dependent IFNγ release, upregulation of the CD25 marker of T cell activation, and target cell lysis (Figures 32A-32C).

[0345] Therapies that strongly involve NK cells may have advantages over therapies that exclusively involve T cells. NK cells do not require antigen priming for activation, do not secrete high levels of cytokines that can induce CRS, and do not induce a graft-versus-host (GVH) response. However, NK cells may require two or more activation stimuli to exert their maximum killing capacity and overcome inhibition by many negative regulatory receptors expressed on NK cells. Furthermore, NK cells are heterogeneously distributed in the TME, and the number of infiltrating NK cells can vary depending on the indication. In some indications, a dysfunctional subset of NK cells has been observed to enrich the TME and correlate with an unfavorable prognosis. Mechanisms that improve NK cell performance may include the simultaneous binding of synergistic NK cell activating receptors, as described herein for CLN-619. In addition, combination therapy with therapies that enhance NK cell function, persistence, and / or tumor infiltration may be beneficial and should be further explored.

[0346] In summary, the data reported herein support the clinical investigation of CLN-619 for the treatment of various tumor types. Phase I clinical trials of CLN-619 as monotherapy and in combination with pembrolizumab in cancer patients are described in further detail below (NCT05117476).

[0347] Example 2. Phase 1 dose-escalation study to investigate the safety, efficacy, pharmacokinetics, and pharmacodynamic activity of CLN-619 (anti-MICA / B antibody) in patients with progressive solid tumors.

[0348] Background: This embodiment describes an ongoing Phase 1, multicenter, open-label, first-in-human dose-escalation study (NCT05117476) evaluating the use of CLN-619 as monotherapy for patients with advanced solid tumors that have progressed to, are intolerant to, or have refused non-curative standard treatments. CLN-619 is a humanized MICA / B-specific IgG1 monoclonal antibody that binds to NKG2D ligands MICA and MICB, preventing proteolytic cleavage from their tumor cells and thereby increasing tumor cell lysis by innate and adaptive immune cells. The amino acid sequence of CLN-619 is shown in Table 1 herein. CLN-619 is expected to have broad antitumor activity.

[0349] Methods: This study was designed to characterize the safety, tolerability, dose-limiting toxicity, and preliminary antitumor activity of intravenous CLN-619 monotherapy in patients with advanced solid tumors. Patients aged 18 years or older with metastatic or locally advanced solid tumors that had progressed after prior treatment and for which no further standard treatments were available, with an ECOG score of 0 or 1, and an estimated life expectancy of 12 weeks or more, were enrolled if they presented with measurable disease according to RECIST v1.1 and demonstrated adequate hepatic and renal function and hematological parameters.

[0350] Patients were excluded from the study if they (i) had received investigative therapy within 25 days (or 5 half-lives) prior to the initial scheduled dose of CLN-619, (ii) had a serious and / or uncontrolled medical impairment, including active autoimmune disease requiring immunosuppressants, (iii) had received treatment for an acute infection within 7 days prior to the initial scheduled dose of CLN-619, (iv) had a grade 3 or higher immunological AE in prior checkpoint inhibitor treatment, or (v) had active CNS metastases.

[0351] CLN-619 was administered intravenously over 1 hour every 3 weeks (Q3W) at dose levels of 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 6 mg / kg, or 10 mg / kg (DL).

[0352] Standard premedication (e.g., acetaminophen or ibuprofen) was administered 30–60 minutes prior to each dose of CLN-619, and before and after each dose according to facility practice.

[0353] Corticosteroid premedication to prevent infusion-associated reactions (IRRs) was required starting at 3 mg / kg DL.

[0354] Parallel dose escalation is being explored for CLN-619 (DL ≥ 1.0 mg / kg) in combination with pembrolizumab (as outlined below in Example 2).

[0355] Response (RECIST 1.1) was assessed every 9 weeks. Patients were allowed to continue therapy for disease progression, unacceptable toxicity, or up to 34 cycles. Patients with disease progression may continue treatment if clinical benefit and acceptable tolerability are determined.

[0356] The study evaluation items include adverse events (CTCAE v5.0), dose-limiting toxicity (DLT), response (RECIST v1.1), and PK parameters (e.g., C max AUC 0-504h , t 1 / 2 ) are some examples.

[0357] Data from dose escalation as monotherapy are described herein.

[0358] Results: Patient baseline characteristics are shown in Figure 14, and the number of patients at each dose level is shown in Figure 15. Briefly, 37 patients (median age, 63 years, range 26–83) were enrolled, 62% were female, and 60% had an ECOG PS of 1. Tumor types included colorectal cancer (6), cervical cancer (5), NSCLC (5), sarcoma (4), endometrial cancer (3), prostate cancer (3), ovarian cancer (2), breast cancer (1), duodenal cancer (1), salivary gland adenoid cystic carcinoma (1), renal cell carcinoma (1), melanoma (1), pancreatic cancer (1), parotid gland cancer (1), peritoneal mesothelioma (1), and thyroid cancer (1). All patients had received prior systemic treatment (median 3, range 1–7), and 20 patients had received prior checkpoint inhibitor treatment. The median number of CLN-619 cycles administered was 2 (range, 0-13).

[0359] Therapeutic adverse events (TEAEs) were monitored, and it was observed that CLN-619 was well-tolerated, with the majority of TEAEs being only Grade 1 / 2. TEAEs observed in more than 10% of patients (shown in Figure 16) included abdominal pain, nausea, fever, infusion-associated reactions (IRRs), decreased appetite, fatigue, vomiting, and back pain. IRRs occurred at dose levels of 0.3 mg / kg or higher. All IRRs occurred in the first cycle and resolved within 3 hours of onset. One case of Grade 3 laryngeal edema occurred at a 10 mg / kg DL in the absence of required steroid premedication, leading to discontinuation of the drug. No fatal TEAEs were reported, and none of the adverse events met the DLT criteria defined in the protocol. These data demonstrate the safety of CLN-619 in a clinical setting. As shown in Figures 17A and 17B, C max A nearly dose-proportional increase in CLN-619 exposure, as measured by AUC, was observed at dose levels from 0.1 mg / kg to 10 mg / kg. 0-504h A greater-than-dose-proportional increase in CLN-619 exposure was observed at dose levels of 0.1–1 mg / kg, as measured by [method / tool], while a nearly dose-proportional increase in CLN-619 exposure was observed at dose levels of 1–10 mg / kg.

[0360] The half-life of CLN-619 ranged from 61.3 to 443 hours for DL ​​doses in the range of 0.1 mg / kg to 10 mg / kg.

[0361] Cytokine expression in longitudinal serum samples from patients treated with CLN-619 monotherapy was examined using Myriad RBM Inflammation and a custom MAP on the Luminex platform. The data are shown in Figure 18, which displays the mean absolute cytokine levels (pg / ml) per dose level over time. Error bars indicate standard deviations (SD). As shown in Figure 18, a transient increase in cytokine levels was observed 2–6 hours after the first dose of CLN-619, decreasing to baseline by days 4–8. No dose-dependent increase in cytokine secretion was observed in the periphery.

[0362] The duration of treatment and clinical activity are shown in Figures 19A and 19B. A summary of objective responses and the best monotherapy efficacy are outlined in Figures 20A and 20B, respectively. A confirmed complete response was observed in a patient with recurrent salivary gland tumor that had progressed after complete response with a PD-1 blocker (Figure 21). The data shown in Figure 22 confirm the objective response in a patient with endometrial cancer and a high tumor burden that had progressed after platinum chemotherapy and anastrazole treatment. Disease stabilization for more than 3 cycles was observed in 7 other patients. For example, one patient with cervical cancer maintained SD for 9 cycles before progression.

[0363] Conclusion: CLN-619 therapy was well-tolerated at doses ranging from 0.1 to 10 mg / kg, and no dose-limiting toxicities were observed at any of the administered doses, thereby demonstrating the monotherapy clinical activity of CLN-619. Furthermore, objective responses were observed in patients who progressed after PD-1. No grade 3 or higher TRAEs were observed in patients who received the premedication mandated by the protocol. Monotherapy activity, including objective responses, was observed across multiple tumor types in both checkpoint-experienced and checkpoint-inexperienced patients. Due to the observation of remarkable monotherapy clinical activity in multiple gynecological malignancies and the demonstration of remarkable efficacy in these subjects, expanded cohorts were established for endometrial cancer and cervical cancer.

[0364] Example 3. Phase 1 dose-escalation study to investigate the safety, efficacy, pharmacokinetics, and pharmacodynamic activity of CLN-619 (anti-MICA / MICB antibody) alone and in combination with pembrolizumab in patients with advanced solid tumors.

[0365] The above examples demonstrate the remarkable and unexpected safety and efficacy of anti-MICA / B antibodies as monotherapy, a pan-cancer treatment. These examples further illustrate the potential of this treatment in combination therapy. Currently, there are no approved formulations targeting the NKG2D ligand / receptor pathway. CLN-619 targets this critical pathway, representing a novel approach in immuno-oncology that could benefit cancer patients in the future.

[0366] Study Description: A phase 1, open-label, first-in-human, multicenter, dose-escalation and dose-expansion study of CLN-619 administered alone (Modules A and D), in combination with pembrolizumab (Module B), or in combination with chemotherapy (Module C) in selected patients with advanced solid tumors. See Figures 23A and 23B for the overall study design.

[0367] CLN-619 will also be administered weekly during the first cycle to study the loading dose schedule.

[0368] Pembrolizumab is an anti-PD-1 antibody administered to patients as a 200 mg Q3W intravenous infusion over 30 minutes.

[0369] The following are examples of chemotherapy regimens that may be administered:

[0370] Carboplatin AUC5 and pemetrexed 500 mg / m2 Q3W IV,

[0371] Carboplatin AUC5-6 Q3W and paclitaxel (175 mg / m2 Q3W) IV,

[0372] Paclitaxel (60-80 mg / m2 Q1W) IV.

[0373] Clinical research:

[0374] As of June 22, 2023, CLN-619 is the only antibody targeting MICA / MICB that has progressed to clinical trials. In the current study, the first-in-human trial of CLN-619, multiple doses of CLN-619 (maximum dose of 10 mg / kg) were administered every three weeks to 61 patients, either alone or in combination with pembrolizumab. CLN-619 was well-tolerated in both monotherapy and combination therapy settings, no dose-limiting toxicities were observed during dose escalation, and no grade 3 treatment-related adverse events (Ae) were observed in patients who received protocol-mandated premedication. At the time of analysis, a total of 235 doses of CLN-619 had been administered, 52 of which were in combination with pembrolizumab. Expanded monotherapy cohorts have been initiated and are currently ongoing in cervical and endometrial cancer.

[0375] Pembrolizumab. Marketed as Keytruda® in the United States, pembrolizumab is a humanized IgG4 anti-PD-1 monoclonal antibody manufactured by Merck for the treatment of cancer. Pembrolizumab acts as an immunomodulatory molecule by blocking the interaction between the programmed cell death 1 (PD-1) receptor in activated T cells and its ligands, PD-L1 and PD-L2, which are expressed on tumor cells and immune cells.

[0376] The rationale for CLN-619 in combination with pembrolizumab

[0377] There is strong rationale for modulating the targeting of both innate and adaptive immunity using the combination of CLN-619 and pembrolizumab. CLN-619 can enhance NKG2D-mediated cell death by both NK cell populations and selected T cell populations, and can also enhance antibody-dependent cell-dependent cytotoxicity, a key NK cell-mediated cytolytic mechanism among other innate immune cell populations. On the other hand, pembrolizumab can enhance T cell-mediated immunity by inhibiting T cell exhaustion mediated by PD-1 and its ligands, PD-L1 and PD-L2. More recently, evidence suggests that the PD-1 / PD-L1 axis may also play a role in regulating NK cell function (Dunai and Murphy 2018). Therefore, through their respective mechanisms of action, both CLN-619 and pembrolizumab can enhance aspects of both innate and adaptive immunity.

[0378] In a clinical setting, data suggest a potential synergistic effect between CLN-619 and checkpoint inhibitors (CPIs). For example, in melanoma patients treated with CPIs, those with detectable levels of sMICA / sMICB had reduced overall survival (OS) compared to those negative for sMICA / sMICB (Maccalli 2017). Therefore, if CLN-619 treatment stabilizes MICA / MICB levels on the tumor surface, CPI therapy may be more effective. It has also been hypothesized that sustained NKG2D signaling can lead to a PD-L1-rich tumor microenvironment, suggesting that NKG2D modulating agents may benefit from combination therapy with PD-1 pathway inhibitors (Sheppard 2018).

[0379] chemotherapy

[0380] Several types of chemotherapy exist with different mechanisms of action. The types of chemotherapy administered during this study are briefly described below.

[0381] Platinum agents (carboplatin). Platinum compounds have two main mechanisms of action regarding cytotoxic effects. They are alkylating agents that bind to and crosslink DNA strands, thereby inhibiting DNA synthesis and function. When DNA is sufficiently damaged, cells undergo apoptosis. Platinum chemotherapy is also immunogenic and can induce a type of cell death that is independent of DNA binding. They interact with many cellular proteins, thereby modulating several signaling pathways.

[0382] Taxane (paclitaxel). Taxanes exert their anticancer effects by binding to the microtubule spindle mechanism, preventing their depolymerization. This results in the inhibition of the transition from metaphase to anaphase, ultimately leading to inhibition of mitosis and induction of apoptosis. Unlike other microtubule inhibitors (e.g., vinca alkaloids), paclitaxel specifically stabilizes microtubules by binding to polymer tubulin, thereby preventing tubulin degradation. The broad-spectrum activity of paclitaxel was predicted by this mechanism of action, which targets very fundamental elements of the cancer phenotype (i.e., regulation of cell proliferation and DNA repair).

[0383] Pemetrexed. Pemetrexed is a novel multi-targeted folate antagonist that inhibits enzymes involved in folate metabolism and purine and pyrimidine synthesis. These precursors interfere with DNA and RNA formation, which are necessary for the growth and survival of both normal and cancer cells.

[0384] The rationale for using CLN-619 in combination with chemotherapy.

[0385] There is strong rationale for combining CLN-619 with standard chemotherapy agents. Cytotoxic chemotherapy, such as platinum, taxanes, and folic acid antimetabolites, has been demonstrated to induce cellular stress that leads to upregulation of NKG2D ligands, including MICA / B. In addition, these agents have been shown to positively influence the immune response, including increased immune cell infiltration and effector cell activity, while simultaneously reducing the levels and function of suppressor immune cells. Due to their respective mechanisms of action, CLN-619 and chemotherapy agents may have synergistic effects.

[0386] Platinum-based agents such as carboplatin and cisplatin have been shown to amplify MICA / B expression, as demonstrated preclinically in small studies of carboplatin-treated ovarian cancer patients and in cisplatin-treated NSCLC cell lines (Capellero 2020, Okita 2019). These agents have also been shown to positively modulate the immune response in preclinical models and, in some cases, clinically. In preclinical models, cisplatin treatment resulted in the recruitment / proliferation of effector cells, upregulation of the lytic activity of cytotoxic effectors, and a reduction in Tregs in TMEs (Gameiro 2012, de Biasi 2014). While no effect on NK cells was observed with carboplatin treatment, carboplatin treatment of ovarian cancer patients resulted in an improved ability of CD8+ T cells to produce IFNγ in ex vivo studies (Wu 2009).

[0387] Taxane chemotherapy, such as docetaxel, has been shown to upregulate MICA / B expression, and the NKG2D-MICA pathway has been demonstrated to be important for docetaxel-mediated NK cell lysis in preclinical models (Acebes-Huerta, 2015). These drugs have also been shown to have a positive effect on immune function in preclinical models and patients. In preclinical models and breast cancer patient samples, docetaxel treatment has been shown to increase NKG2D in NK cells, and in preclinical models, docetaxel mediated an increased responsiveness to trastuzumab via NKG2D (Di Modica 2016). Increased tumor infiltration of CD8+ T cells was observed in NSCLC patients treated with docetaxel (Gao 2019). Finally, paclitaxel was demonstrated to reprogram tumor-associated macrophages to a phenotype closer to M1 type or more immunogenic in both preclinical and patient samples (Wanderley 2018).

[0388] Although the folate antagonist pemetrexed has not been shown to induce MICA / B expression, it has been shown to have a positive effect on T cell activity (Okimoto 2020, Schaer 2019). In preclinical studies, pemetrexed increased T cell activation in mouse tumors in vivo, strongly induced immunogenic cell death in mouse tumor cells, and enhanced T cell activation in vitro (Schaer 2019). Furthermore, it has been shown to induce PD-L1 expression in NSCLC cells, and when used in combination with a PD-1 blocker, it resulted in significantly enhanced antitumor activity in a preclinical model (Schaer 2019).

[0389] In addition to the data above highlighting the role of chemotherapy in positively modulating both the MICA-NKG2D pathway and immune effector activity, immunotherapy-assisted chemotherapy has been widely tested in conjunction with anti-PD(L)1 therapy in multiple tumor types. Pembrolizumab, used in combination with platinum- and taxane-based chemotherapy, is approved for multiple tumor types, including NSCLC, endometrial cancer, breast cancer, and gastric cancer (pembro uspi 2021). Dostallimab, also used in combination with platinum-based chemotherapy, is approved as a first-line treatment for patients with mismatch repair deficiency (dMMR) advanced endometrial cancer (N Engl J Med 2023;388:2145-58.DOI:10.1056 / NEJMoa2216334). While combination therapy with anti-PD(L)1 chemotherapy has shown success in some solid tumors, its efficacy has not been demonstrated in other solid tumors, such as epithelial ovarian cancer or NSCLC driven by targetable factors, such as NSCLC with EGFRm or ALK rearrangements.

[0390] Cytotoxic chemotherapy can produce immunomodulatory effects, such as blocking immunosuppressive pathways and enhancing cytotoxic T cell responses. Therefore, the combination of chemotherapy and immunotherapy may have synergistic effects in the tumor microenvironment. Clinical benefits, including improved survival, have been reported for this combination in several cancer types. Considering the mechanisms of action of individual cytotoxic agents that induce cellular stress and MICA / B expression, the combination of chemotherapy and CLN619 may be synergistic, and there is strong clinical rationale to explore in this study.

[0391] Rational basis for starting dose and schedule

[0392] The first-in-history (FIH) dose and dosing regimen for CLN-619 were calculated using allometric scaling to estimate human PK and expected receptor occupancy (RO), in addition to in vivo and in vitro safety and pharmacological data. Using this method, a starting dose of 0.1 mg / kg and a Q3W dosing schedule are proposed for the CLN-619 clinical trial. Based on the overall available toxicological and pharmacological data, the starting dose is expected to be safe but within the lower range of CLN-619's pharmacological activity.

[0393] Human PK parameters were determined based on allometric scaling from cynomolgus monkey PK data. The expected clearance in humans is calculated to be 0.19 L / day. The predicted terminal phase half-life in humans is 26.9 days, which is typical for monoclonal antibodies and justifies the selection of a Q3W dosing schedule.

[0394] The NOAEL (No Observed Adverse Emission Limit) (101.4 mg / kg / week) in a 1-month repeated dose monkey toxicity study supports 0.1 mg / kg as a safe starting dose. At the proposed starting dose of 0.1 mg / kg, after a single dose of CLN-619, the area under the predicted concentration curve (AUC0-168) and Cmax are 1 / 775th and 1 / 1020th, respectively, of the AUC0-168 and Cmax at the maximum dose (equal to the NOAEL) in cynomolgus monkeys where serious toxicity does not occur, providing a broad safety margin.

[0395] The lack of findings observed after administration of CLN-619 to monkeys is not inconsistent with the biological nature of the target. Notably, MICA / MICB expression in normal tissues is highly restricted. MICA / MICB expression is induced under stress conditions (e.g., infection, radiation, malignancy), which is a normal physiological process that triggers the elimination of MICA / MICB-expressing cells by immune cells. Since high levels of normal tissue expression of MICA and MICB are not expected in healthy monkeys, the effects of CLN-619 treatment on normal animals are expected to be minimal.

[0396] Using modeling techniques, the predicted RO of CLN-619 to MICA and MICB was determined using in vitro association rate constants and equilibrium dissociation constants. The total target occupancy of MICA / MICB is predicted to be approximately 30% at the starting dose of CLN619. Based on the intended administration of CLN619 to late-stage cancer patients, where the objective is to start with a pharmacologically active dose (PAD) considering patient safety, a predicted RO of 30% is considered reasonable.

[0397] In determining the starting dose of CLN-619, in vivo and in vitro pharmacological data were also considered. Based on both in vivo and in vitro pharmacological data, the proposed starting dose of CLN-619 is expected to be within the lower range of pharmacological activity. At the proposed starting dose of 0.1 mg / kg, administered as a 1-hour infusion, the expected Cavg in humans is approximately one-fifth of the measured Cavg achieved at effective doses in mice in in vivo xenograft mouse studies. In in vitro functional studies measuring CLN-619 target-modulating function and Fc effector function in tumor cell lines, EC50 values ​​ranged from 0.045 to 0.1 μg / mL. However, caution should be exercised in the relationship between in vitro data and FIH dose determination, considering that tumor exposure levels are predicted to be one-tenth of circulating levels, which could lead to an overestimation of efficacy in tumors. In addition, while in vitro functional assays are technically effective in demonstrating mechanism-driven activity, the sponsor believes that the usefulness of these evaluations is limited because in vitro assays are closed and static systems and do not represent the open and dynamic in vivo conditions in which CLN-619 is continuously eliminated over time.

[0398] In vitro cytokine release assays further support the expectation that the starting dose is pharmacologically active and safe. Treatment of human PBMCs (6 donors) with CLN-619 failed to induce cytokine / chemokine responses above background levels at all test concentrations, in both soluble and wet-binding formats. In the context of co-culture systems of PBMCs and MICA / MICB-expressing cell lines, CLN-619 induced the release of a limited set of cytokines, including MIP1α, TNFα, and IL-8, which are considered to be appropriate pharmacodynamics. In the co-culture system, TNFα levels across the 6 donors were 1 / 16 to 1 / 210 of those of the positive control at concentration levels closest to the predicted Cmax in humans. Overall, based on the in vitro human cytokine release data and the lack of cytokine release in a 1-month central in vivo primate toxicity study, the risk of cytokine release at the clinical starting dose of CLN619 is low.

[0399] Based on this data, we selected a starting dose of 0.1 mg / kg and a Q3W dosing schedule, which we anticipated would be safe but within the lower range of CLN-619's pharmacological activity.

[0400] A preliminary analysis of CLN-619 PK data collected for dose levels ranging from 0.1 mg / kg to 10 mg / kg indicates that the increase in exposure is proportional to the dose, at least in the range of 1 mg / kg to 10 mg / kg. Therefore, at these dose levels and higher, the PK of CLN-619 is linear, and consequently, the increase in total exposure is considered to be analogous to the increase in total dose. At higher dose levels, the mean half-life can be as long as 19 days, indicating that steady-state exposure may not be achieved for 60–90 days or approximately 3–6 cycles of CLN-619.

[0401] To achieve steady-state exposure more rapidly and limit the maximum serum concentration of CLN-619, a loading dose strategy can be used in the first cycle. The planned loading dose regimen involves administering 10 mg / kg of CLN-619 three times (3 times) on a weekly schedule in the first cycle. An additional 10 mg / kg dose is administered on day 1 of the second cycle, and from the second cycle onward, CLN-619 is administered on a Q3W schedule. As described above, the overall increase in dose is directly proportional to the overall increase in exposure. Therefore, in the first cycle, the overall exposure (AUC) will increase by approximately three times compared to the first cycle on a 10 mg / kg Q3W schedule. However, during this period, the overall exposure is not expected to significantly exceed the steady-state exposure observed on a 10 mg / kg Q3W schedule, which has been shown to be well-tolerated. By using a loading dose, steady-state exposure is expected to be achieved by the end of the first cycle, which may offer a higher chance of achieving a rapid response.

[0402] Given that the load dose is not expected to achieve a higher systemic exposure than previously observed at steady state with the current 10 mg / kg Q3W, this dose increase is considered safe.

[0403] Safety and risk / benefit assessment

[0404] As of June 22, 2023, 61 patients had been exposed to CLN-619 doses of 0.1, 0.3, 1, 3, 6, and 10 mg / kg. 43 patients were exposed to CLN-619 monotherapy, with 12 receiving the 10 mg / kg dose. 18 patients were exposed to CLN-619 in combination with pembrolizumab at doses of 1, 3, 6, and 10 mg / kg, with 6 receiving the 10 mg / kg dose. A total of 23 treatment-induced SAEs were reported after administration of CLN-619 (in monotherapy and in combination with pembrolizumab). A single patient reported a serious infusion-associated reaction (IRR) with laryngeal edema related to CLN-619 in the absence of mandatory steroid premedication. One non-serious grade 3 rash was reported in monotherapy (module A). In combination therapy (module B), no serious adverse reactions to CLN-619 were reported; however, one SAE of acute kidney injury (AKI) was anticipated and thought to be potentially related to pembrolizumab (rather than CLN-619). None of the Ae met the dose-limiting toxicity (DLT) criteria defined in the protocol, and no grade 4 or higher TEAEs were observed. The most common treatment-related adverse events (TEAEs) observed to date in 5% or more of patients treated with CLN-619 monotherapy were IRR (23.3%), abdominal pain (18.6%), fatigue (18.6%), and fever (18.6%). In patients treated with CLN-619 in combination with pembrolizumab, the most common adverse events observed to date in more than 5% of patients were constipation (22.2%), anemia (16.7%), back pain (16.7%), elevated serum creatinine (16.7%), fatigue (16.7%), and nausea (16.7%). The latest version of the Investigational Brochure (IB) provides more detailed safety information.

[0405] The estimated frequency of IRR associated with the first dose was 21.3% (13 occurrences in 61 treated patients). The estimated frequency across all doses was 6.3% (15 occurrences in 235 CLN-619 administrations). The majority of IRRs (13) occurred during or immediately after the first dose, one case of IRR occurred after the sixth dose, and one patient experienced IRR both after the first and second doses. One event occurred in a patient receiving concomitant therapy with pembrolizumab. The number of patients exposed to concomitant therapy remains limited (18), and therefore, it is difficult to assess whether this addition alters the risk of IRR. All patients recovered without sequelae, but IRR can potentially be severe in some patients.

[0406] A 60-minute infusion time and IV administration were initially selected for this study, supported by a nonclinical toxicology study in which CLN-619 was administered weekly as a 5-minute slow IV bolus injection. Based on clinical experience in the dose-escalation cohort, the infusion time was extended to 120 minutes for the first one to two infusions to reduce the frequency and severity of IRRs.

[0407] CLN-619-001 is the first-in-human clinical trial of CLN-619 administered monotherapy and in combination with pembrolizumab (anti-PD-1 antibody) in patients with locally advanced or metastatic solid tumors. Patients in the Module B cohort must have the tumor type described in the current prescribing information for pembrolizumab. Restrictions regarding prior treatment history are described in the inclusion criteria. Unless otherwise specified in the inclusion criteria, patients must have received, declined, or demonstrated contraindications or intolerances to any approved standard therapy available to them. CLN-619-001 utilizes a modular study design, with Module A designed to investigate the safety, tolerability, and preliminary antitumor activity of CLN-619 monotherapy, and the two-part design supporting Module B, which investigates CLN-619 in combination with pembrolizumab (anti-PD-1 antibody) (see Figure 23B). Module A monotherapy dose escalation will evaluate the safety of CLN-619 at multiple dose escalation levels to define the maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D) of CLN-619 monotherapy. Assuming acceptable safety and tolerability of CLN-619 monotherapy in dose escalation, Module A cohort expansion will be initiated to enroll tumor-specific cohorts of patients with non-small cell lung cancer (NSCLC), cervical cancer, and endometrial cancer. Based on evidence of clinical utility, and / or any observed relationships between blood and tumor-based biomarker data and objective responses or other applicable new data observed during the study, up to three additional cohorts of patients with specific tumor types may be enrolled in Module A cohort expansion. These cohorts will further evaluate the PK / pharmacodynamics (PD), safety, and initial antitumor activity of CLN-619 monotherapy.

[0408] The two-part study design also supports the initiation of a Module B combination therapy dose-escalation cohort in patients with advanced solid tumors. This cohort will investigate the safety, tolerability, and initial antitumor activity of CLN-619 in combination with pembrolizumab using a 3+3 design. Once the safety of the 3 mg / kg monotherapy dose is established, the Module B combination therapy dose-escalation cohort may be initiated with a 1 mg / kg CLN-619 dose. If the monotherapy CLN-619 MTD is less than 3 mg / kg, the Module B combination therapy dose-escalation cohort may be initiated at a dose level one step lower than the MTD. Pembrolizumab (200 mg) will be administered immediately before CLN-619. A standard 3+3 dose-escalation scheme will be followed. Dose levels of 1.0, 3.0, 6.0, and 10 mg / kg are planned.

[0409] Following the demonstration of acceptable safety and tolerability of the combination of CLN-619 and pembrolizumab, the Module B cohort expansion will evaluate the preliminary safety and antitumor activity of the combination of CLN-619 and pembrolizumab in patients with NSCLC and endometrial cancer. Based on evidence of clinical utility, and / or any observed relationships between blood and tumor-based biomarker data, observed objective response or disease stabilization, or other applicable new data, up to three additional disease-specific expansion cohorts may be enrolled in the Module B cohort expansion. The monotherapy dose-escalation cohort and the combination therapy dose-escalation cohort will enroll patients cautiously and stepwise under strict monitoring, based on standard decision rules.

[0410] Module C explores the safety, tolerability, and preliminary efficacy of combination standard chemotherapy in predefined tumor types. The combination dose expansion cohort will enroll patients cautiously and stepwise under strict monitoring, based on standard decision rules.

[0411] By adding Module D, the sponsor will investigate whether the weekly loading dose in the first cycle can achieve steady-state exposure by the end of the first cycle and offer a higher probability of achieving a rapid response. Given that the loading dose is not expected to achieve higher systemic exposure than previously observed at steady state with the current 10 mg / kg Q3W, which has been shown to be well-tolerated, this dose increase is expected to be safe. Taken together, this study provides a reasonable risk:benefit assessment for patients with advanced cancer who have few treatment options.

[0412] Module A monotherapy dose escalation

[0413] A selected cohort of patients with advanced solid tumors will be treated with escalating doses of CLN-619 monotherapy, initially using a single-patient accelerated titration design. If the first patient enrolled at a given dose level experiences a grade 2 or higher CLN-619-related (defined as potentially related or clearly related) adverse event (AE) during the first three-week treatment cycle or at the 3.0 mg / kg dose level, whichever comes first, a transition to a standard 3+3 dose escalation scheme will be initiated. The planned dose levels are shown in Table 3 below. [Table 3]

[0414] Use the data obtained during Module A dose escalation to identify the MTD (or MAD if MTD is not defined) for CLN-619 monotherapy. Alternatively, based on a review of PK / PD, safety, and efficacy data, a MBED (Mean Basic Efficacy Dose) below the MTD may be designated, which can also guide dose selection for expanding the Module A cohort (i.e., RP2D).

[0415] Module A monotherapy dose escalation, dose-level cohort extension

[0416] The purpose of extending enrollment in these dose-level cohorts is to further characterize the safety, PK, PD, biomarkers, and clinical activity of CLN-619, as well as to characterize the clinical and bioactivity of the selected dose levels for extension.

[0417] For enrollment in each cohort extension at the selected dose level, we planned for up to 10 patients with advanced solid tumors, including those enrolled in the initial dose-level cohort. A total of 45 patients were enrolled in the seven cohorts.

[0418] The study procedures for cohort extension are identical to those in the original cohort, as described in the event schedule (Figure 24) and sampling schedule (Figure 25), except that two essential fresh biopsies are taken at baseline (screening) and after the first cycle of medication (i.e., C2D8 -1 day / +3 days).

[0419] Module A monotherapy cohort expansion

[0420] Once RP2D for CLN-619 monotherapy is selected, Module A monotherapy cohort expansion will be initiated. This module will enroll the following tumor-specific expansion cohorts to investigate the safety and initial antitumor activity of CLN-619 monotherapy: ● Expanded A1: CLN-619 monotherapy in patients with non-small cell lung cancer (NSCLC) (N=16 patients). ● Enlarged A2: CLN-619 monotherapy in patients with cervical cancer (N=16 patients). ● Expanded A3: CLN-619 monotherapy at dose level #1 in patients with endometrial cancer (N=10 patients) ● Enlarged A4: CLN-619 monotherapy at dose level #2 in patients with endometrial cancer (N=10 patients) ● Based on evidence of clinical benefit in that tumor subtype during the dose escalation phase of the study (objective response or disease stabilization lasting at least 6 cycles of treatment), up to three additional disease-specific expansion cohorts may be established. Alternatively, if these additional tumor-specific expansion cohorts are established, the relationship between blood and tumor-based biomarker data and objective response may be considered. ● Expanded A1 and A2: If two out of 16 patients experience clinical benefit (defined as an objective response or disease stability sustained over at least six cycles of treatment), the cohort may be further expanded to enroll up to a total of 40 patients to further characterize the preliminary antitumor activity of CLN-619 monotherapy in that population. ● Expansion A3 and A4: Each cohort will initially be expanded to 10 patients, including those who received medication at each dose level during dose escalation. If at least one objective response is observed in the first 10 treated patients, additional patients may be enrolled until the total number of patients with endometrial cancer reaches up to 40 (20 at each dose level, including those who received medication during the dose escalation phase). ● Additional disease-specific expansion cohorts: Each cohort will initially be expanded to 10 patients. If evidence of clinical benefit (objective response or disease stabilization lasting at least 6 cycles of treatment) is observed in at least one of the first 10 treated patients, additional patients may be enrolled until there are a total of up to 40 patients with a defined tumor type (including patients treated during the dose escalation phase).

[0421] Module B combination therapy dose escalation

[0422] Module B combination therapy dose escalation will be initiated to investigate the safety, tolerability, and initial antitumor activity of CLN-619 in combination with pembrolizumab. These cohorts will enroll patients with the tumor type described in the current prescribing information for pembrolizumab. The cohort may be enrolled starting at dose level 1 (1 mg / kg), in parallel with the Module A monotherapy expansion cohort, if the safety of the 3 mg / kg monotherapy dose level has been demonstrated. If a lower dose is determined to be the MTD in the Module A monotherapy dose escalation group, dose levels below the MTD will be investigated in combination with pembrolizumab.

[0423] The planned dose levels are shown in Table 4 below. [Table 4]

[0424] For the first three patients at dose level 1, if no DLT is observed during the first three-week cycle, CLN-619 should be escalated. If a DLT is present as indicated in the decision rules, tapering may be performed at the dose levels listed in the table.

[0425] Module B combination therapy cohort expansion

[0426] Once the acceptable safety and tolerability of CLN-619 in combination with pembrolizumab are demonstrated, Module B cohort expansion will be initiated. This module will investigate the safety and initial antitumor activity of CLN-619 in combination with pembrolizumab by enrolling patients with NSCLC, endometrial cancer, and up to three additional tumor subtypes. Module B cohort expansion can only be initiated after six patients have been enrolled at the same dose level and the safety of that dose level has been demonstrated based on the 3+3 design decision rule. ● Expanded B1: CLN-619 + pembrolizumab in patients with NSCLC (N=16) ● Expanded B2: CLN-619 + pembrolizumab in patients with endometrial cancer (N=16) ● Based on evidence of clinical benefit in that tumor subtype during the study's dose escalation phase (objective response or disease stabilization lasting at least six cycles of treatment), up to three additional disease-specific cohorts may be established. Alternatively, if these additional tumor-specific expansion cohorts are established, the relationship between blood and tumor-based biomarker data and objective response may be considered. ● Expanded B1 and additional disease-specific expanded cohorts: If two out of 16 patients experience clinical benefit (defined as an objective response or disease stabilization lasting at least six cycles of treatment), the cohort may be further expanded to enroll up to a total of 40 patients (including those treated during the study's dose escalation phase) to further characterize the safety and preliminary antitumor activity of CLN-619 monotherapy in that population. ● Expansion B2: Patients are alternately assigned to receive one of two dose levels of CLN-619 in combination with pembrolizumab. If two of the 16 patients experience clinical benefit (defined as an objective response or disease stability that lasts for at least six cycles of treatment), the cohort may enroll up to a total of 40 additional patients.

[0427] Module C: CLN-619 + chemotherapy combination therapy, escalation and expansion.

[0428] Module C Dose Elevation: Each chemotherapy cohort will be investigated at doses of 3 mg / kg and 10 mg / kg. Patients will be enrolled in CLN-619 at 3 mg / kg in combination with standard dose chemotherapy, based on the 3+3 design. For each tumor type, CLN-619 + chemotherapy will be initiated at 3 mg / kg (n=3-6) and escalated to 10 mg / kg (n=3-6) once safety is confirmed. If a dose limit (DLT) occurs at 10 mg / kg, a lower dose will be selected and the safety process will be repeated. Safety data at each dose level tested in combination with chemotherapy for each tumor type will be reviewed to determine the optimal dose, and the remaining patients will be enrolled in dose escalation, up to a total of 40 patients.

[0429] Module C Disease-Specific Dose Expansion: Initially, 16 patients are enrolled in each cohort. If two of the 16 patients experience clinical benefit (defined as an objective response or disease stabilization sustained over at least six cycles of treatment), the cohort may be further expanded to enroll up to a total of 40 patients to further characterize the safety and preliminary antitumor activity of CLN-619 in combination with chemotherapy in that population. ● Cohort C1: CLN619 + carboplatin + pemetrexed in recurrent EGFRm NSCLC ● Cohort C2: CLN619 + Carboplatin + Paclitaxel in patients with recurrent endometrial cancer ● Cohort C3: CLN619 + Paclitaxel in Platinum-Resistant Epithelial Ovarian Cancer ● Each treatment cycle is every 21 days or every 3 weeks (q21d or q3w). [Table 5] [Table 6] [Table 7]

[0430] Module D loading dose cohort

[0431] This module investigates the safety and antitumor activity of CLN-619 monotherapy. A weekly loading dose of 10 mg / kg is administered in the first cycle (QW x 3). Subsequently, 10 mg / kg Q3W is administered in the second cycle and subsequent cycles (following the usual dosing schedule).

[0432] Selection of research group

[0433] Planned number of patients

[0434] When all modules are fully enrolled, the number of patients will reach approximately 640, excluding those who need to be replaced. Module A monotherapy dose escalation enrolled 45 patients with advanced solid tumors. Up to 240 patients with specific tumor types will be enrolled in the Module A monotherapy cohort expansion. If initiated, 25 patients will be enrolled in the Module B combination therapy dose escalation cohort, up to 200 patients in Module B, and up to 120 patients in the Module C CLN-619 + chemotherapy combination therapy, escalation and expansion cohort. Up to 10 patients will be enrolled in the Module D monotherapy loading dose cohort.

[0435] Selection Criteria

[0436] Patients must meet all of the following selection criteria to be eligible to participate in the study. 1) Male or female aged 18 or older. 2) The patient is willing and capable to submit written informed consent and to comply with protocol requirements. Written informed consent and any necessary local authorizations must be obtained from the patient before any protocol-related procedures, including screening assessments, are implemented. 3) Module A monotherapy dose-escalation cohort and Module B combination therapy dose-escalation cohort: Histologically or cytologically confirmed metastatic or locally advanced unresectable solid tumors. For Module B, tumor types are listed as approved indications based on current prescribing information for pembrolizumab. 4) Module A cohort expansion: a) Expanded A1: Histologically or cytologically confirmed metastatic or locally advanced unresectable NSCLC, b) Expanded A2: Histologically or cytologically confirmed metastatic or locally advanced unresectable cervical cancer. c) Expanded A3 and A4: Histologically or cytologically confirmed metastatic or locally advanced unresectable endometrial cancer. d) Eligibility for disease-specific expanded cohorts may be further refined based on clinical, pharmacodynamic, or biomarker data revealed by the study, including histological subtypes, molecular characteristics, or prior treatment exposure. 5) Module B cohort expansion: a) B1: Histologically or cytologically confirmed metastatic or locally advanced, unresectable NSCLC. b) Expanded B2: Histologically or cytologically confirmed metastatic or locally advanced unresectable endometrial cancer. c) Eligibility for disease-specific expanded cohorts may be further refined based on clinical, pharmacodynamic, or biomarker data revealed by the study, including histological subtypes, molecular characteristics, or prior treatment exposure. 6) Module C CLN-619 + chemotherapy combination therapy, escalation and expansion cohort a) Expanded C1: Recurrent NSCLC confirmed histologically or cytologically. b) Expanded C2: Recurrent endometrial cancer confirmed histologically or cytologically. c) Expanded C3: Histologically or cytologically confirmed recurrent platinum-resistant epithelial ovarian cancer. d) Eligibility for disease-specific expanded cohorts may be further refined based on clinical, pharmacodynamic, or biomarker data revealed by the study, including histological subtypes, molecular characteristics, or prior treatment exposure. 7) Module D monotherapy loading dose 10 mg cohort: Patients with recurrent epithelial ovarian cancer, breast cancer, and gastrointestinal (esophageal, gastric, and colorectal) cancer. 8) The following prior treatments: a) The patient should receive this therapy unless any other approved standard therapy available to the patient is contraindicated, intolerable, or the patient has refused it. If the patient has refused such therapy, the patient should be notified and this refusal should be recorded in the medical record. 9) At baseline, the patient must meet the criteria of RECIST v1.1 and have one or more measurable lesions that meet the following conditions: a) Non-lymph node lesions with the longest one-dimensional measurement of 10 mm or more, or lymph node lesions with the shortest one-dimensional measurement of 15 mm or more, b) Lesions that have previously undergone local treatment such as radiotherapy or ablation can also be used as measurable target lesions if progression has been confirmed according to RECIST v1.1 prior to registration and the longest one-dimensional measurement is 10 mm. 10) Performance status of 0 or 1 based on the East Coast Cancer Clinical Trials Group (ECOG) Performance Scale. 11) Estimated average life expectancy of 12 weeks or more. 12) Previous palliative radiotherapy must have been completed 14 days prior to the administration of C1D1. 13) Toxicity associated with prior study treatments, with the exception of alopecia, must have improved to Grade 1 or less according to the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) v5.0. Peripheral neuropathy must be clinically stable or improving and of Grade 2 or less in severity. Patients with chronic but stable Grade 2 toxicity may be permitted enrollment after agreement between the principal investigator and the sponsor. 14) Having sufficient liver and kidney function and hematological parameters within the normal range as defined below: a) Total bilirubin less than 1.5 × ULN. This does not apply to patients with confirmed Gilbert's syndrome, in which case total bilirubin must be less than 3.0 mg / dL and conjugated bilirubin less than 0.5 mg / dL. b) Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels of 2.5 × ULN or less, or 5 × ULN or less in patients with liver metastases. c) Creatinine clearance (CrCl) of 45 mL / min or greater, as measured or estimated using the Cockcroft-Gault formula. d) Hemoglobin levels of 8 g / dL or higher without transfusion for at least two weeks prior to the administration of C1D1. e) 1500 cells / mm³ without growth factor maintenance, for filgrastim (3 days) and pegfilgrastim (14 days). 3 The above absolute number of good pitches, f)75,000 cells / mm 3 The platelet count is as above. 15) Patients in the Module A and Module B dose-escalation cohorts and in Module D must have preserved tissue available for biomarker analysis. The sample is preferably from a sample obtained after the most recent therapy. If preserved tissue (e.g., all tumor blocks have been used up) is unavailable, a fresh biopsy is required. If a biopsy cannot be performed due to an acceptable clinical risk at the discretion of the principal investigator, the sponsor's medical monitor must be contacted and registration approved. a) Patients participating in the Module A and Module D dose-level cohort extension(s) must agree to provide a fresh biopsy at screening and an in-progress biopsy on day 8 of the second cycle. b) Patients in the expanded cohorts of Modules A, B, and C must consent to providing a fresh pre-procedure biopsy and an intra-procedure biopsy on day 8 of the second cycle. If the biopsy cannot be performed due to an acceptable clinical risk at the discretion of the principal investigator, the sponsor's medical monitor must be contacted and registration approved. Paired biopsies are recommended in the Modules A1 and B1 NSCLC cohorts where clinically feasible.

[0437] Exclusion criteria

[0438] A patient will be excluded if they meet any of the following exclusion criteria: 1) You are currently or have previously participated in an intervention study and received the investigational drug within 28 days (or 5 half-lives, whichever is longer) prior to the C1D1 administration. 2) Patients with a concomitant second malignancy (excluding appropriately treated non-melanoma skin cancer, ductal carcinoma in situ, superficial bladder cancer, prostate cancer, or cervical carcinoma in situ) will be excluded unless they were in complete remission three years prior to study enrollment and will not require or are not expected to require additional therapy during study participation. 3) Patients with any active autoimmune disease, or a history of known or suspected autoimmune disease, or a history of a syndrome requiring systemic corticosteroids or immunosuppressants, except for patients with vitiligo, improved childhood asthma / atopic dermatitis, or autoimmune thyroid disorders receiving stable thyroid hormone replacement therapy. 4) A severe, uncontrolled medical impairment that could impair the patient's ability to receive protocol therapy or that could become difficult to control due to complications of this therapy. These criteria include, but are not limited to, the following: a) Uncontrolled airway hyperresponsiveness, b) Type 1 diabetes. Patients with type 2 diabetes are permitted if they are under stable blood glucose control based on the investigator's assessment. c) Uncontrolled, clinically significant lung disease, d) The need for supplemental oxygen to maintain pulse oximetry values ​​above 93%, e) Records of symptomatic congestive heart failure or ejection fraction less than 45%, based on the investigator's assessment. f) Ejection fraction of less than 45% in patients with a history of anthracycline chemotherapy or a history of ventricular dysfunction. Patients with ventricular dysfunction or a history of anthracycline therapy should undergo echocardiography to assess baseline cardiac function. g) A history of unstable angina or myocardial infarction within 6 months prior to the administration of C1D1, h) Unstable cardiac arrhythmia, i) History of ventricular arrhythmias, j) Uncontrolled hypertension: Patients with persistent systolic blood pressure above 150 or diastolic blood pressure above 100 must be documented by the treating physician as consistent with uncontrolled hypertension. k) A history of stroke or cerebral hemorrhage within one year prior to the administration of C1D1, l) Poorly controlled paroxysmal disorder, m) Active diverticulitis within one year prior to the administration of C1D1, n) A recent major surgery or major surgery with unresolved complications that could interfere with research procedures, within three months prior to the administration of C1D1. 5) Treatment with systemic antiviral, antibacterial, or antifungal agents for acute infection within 7 days prior to the administration of C1D1. 6) Having a history of or a positive test result for HIV I / II primary immunodeficiency disease, such as human immunodeficiency virus (HIV). 7) Diagnosed with hepatitis B (positive test result for either hepatitis B surface antigen [HbsAg] or hepatitis B core Ab) or hepatitis C virus (HCV) infection (positive test result for HCV antibody and / or HCV ribonucleic acid [RNA] in serum) under any of the following conditions: a) The patient has active hepatitis B or C and has received antiretroviral therapy within the past four weeks. b) Hepatitis B deoxyribonucleic acid (DNA) or HCV RNA can be detected in the blood. 8) History of allogeneic organ transplantation or allogeneic hematopoietic transplantation. 9) A history of any of the following events accompanied by pretreatment with checkpoint inhibitor immunotherapy: Grade 3 or higher neurotoxicity, ocular toxicity, interstitial pneumonia, myocarditis, or colitis; hepatic impairment meeting the Hy rule criteria. 10) Symptomatic, uncontrolled brain metastases, known or suspected leptomeningeal metastases, and / or carcinomatous meningitis. Patients in whom brain metastases are identified at screening may be screened again after receiving appropriate treatment (e.g., surgical treatment and / or radiotherapy). Patients whose brain metastases have been treated should be neurologically stable for 28 days after completion of treatment and prior to registration, and should receive a stable regimen of steroids (less than 10 mg of prednisone or equivalent) for 14 days prior to C1D1 administration. 11) Treatment with a non-tumor vaccine (i.e., HPV vaccine) for the control of infectious disease within 28 days prior to C1D1. Inactivated seasonal influenza vaccine may be administered to patients without restriction before the start of treatment and during the study therapy. Live virus-containing influenza vaccine or other clinically required vaccines for infectious disease (i.e., pneumovax, varicella) may be permitted, but must be discussed in advance with the sponsor's medical monitor and may require a washout period for the study drug before and / or after vaccine administration. Covid-19 vaccine may be administered according to the institution's policy. 12) Patients with active SARS-CoV-2 infection, including those with a history of positive SARS-CoV-2 testing, who have no record of subsequent negative test results, patients whose results are pending and not yet known, or patients suspected of having an active infection based on clinical characteristics. SARS-CoV-2 vaccination during treatment is permitted. 13) Within 28 days prior to the administration of C1D1 medication, the patient received an immunosuppressant including, but not limited to, CellCept, methotrexate, infliximab, anakinra, tocilizumab, cyclosporine, or a corticosteroid (prednisone or equivalent at 10 mg / day or more). 14) Pregnant or breastfeeding women (FOCBPs) who plan to become pregnant within 120 days of the last dose of the study drug, or who refuse to use an acceptable method to prevent pregnancy during the study procedure and for 120 days after the last dose of the study drug. a) Women of childbearing potential are defined as follows: i) Not surgically infertile, i.e., not having undergone bilateral tubal ligation, bilateral oophorectomy, or hysterectomy, or ii) Not a postmenopausal condition as defined as amenorrhea of ​​two years or more without an alternative medical cause. Note: Women who have had amenorrhea for less than two years and are not surgically infertile (i.e., have not undergone tubal ligation, bilateral oophorectomy, or hysterectomy) are considered fertile only if they have a record of follicle-stimulating hormone (FSH) levels within the postmenopausal range. 15) Male patients who plan to become fathers or donate sperm within 120 days of the last dose of the study drug or within 5 half-lives of CLN-619, whichever is longer, or who have a partner who is a FOCBP and refuse to use any acceptable method to prevent pregnancy during the study procedure and for 120 days after the last dose of the study drug or within 5 half-lives of CLN-619, whichever is longer. 16) A QT interval (QTcF) of 500 milliseconds or longer, corrected for heart rate using the Fridericia formula. 17) The patient has a history of drug-related anaphylactic reaction to any component of CLN-619 (Module A and Module B patients) or pembrolizumab (Module B patients only). A history of grade 4 anaphylactic reaction to any monoclonal antibody therapy. 18) Known ongoing alcohol or drug abuse. 19) Inability to comply with the protocol and / or unwillingness to conduct or access to follow-up evaluations. 20) Patients who lack the capacity to make decisions or who are incarcerated against their will. 21) Patients deemed unsuitable for participation based on the judgment of the principal investigator. 22) Treatment using any of the following: a) Systemic anticancer treatment within 14 days prior to the first dose of the C1D1 research drug. b) Immunotherapy within 28 days prior to the first dose of the C1D1 research drug. c) Radiotherapy within 28 days prior to the first dose of the C1D1 study drug, and palliative radiation within 14 days prior. If irradiated, the lesion must demonstrate clear progression before qualifying as a target lesion. d) Major surgery (excluding vascular access placement) within 28 days prior to the initial administration of the C1D1 research drug.

[0439] Diet and dietary restrictions

[0440] Patients should maintain a normal diet unless adjustments are needed to manage adverse events such as diarrhea, nausea, or vomiting.

[0441] Criteria for discontinuing treatment

[0442] Patients who discontinue the procedure should be encouraged to return for safety follow-up appointments and to participate in survival follow-ups. If a patient discontinues both the procedure and follow-up, no further evaluation or data collection is required.

[0443] Reasons for discontinuing treatment include, but are not limited to, the following: ● Completion of 34 cycles of therapy, ● Disease progression or the need for anti-cancer therapy not specified in the protocol, ● In the opinion of the principal investigator, further treatment would not be in the best interests of the patient due to comorbidities, systemic or local changes in the patient's condition, ● Death ● DLT or unmanageable adverse events related to the therapy, ● Withdrawal of consent regarding the procedure, ● Serious protocol violation or non-compliance, ● The clinical trial sponsor has completed the research.

[0444] For patients who meet the criteria for discontinuation of treatment but otherwise seem to be obtaining attractive clinical benefits from ongoing treatment with CLN-619, pembrolizumab, CLN-619 in combination with pembrolizumab, or CLN-619 in combination with chemotherapy, the continuation of research therapy may be considered on an individual basis.

[0445] Criteria for End-of-Trial (EOT)

[0446] Patients should be encouraged to complete all research evaluations. However, patients may withdraw their consent to participate in this study at any time for any reason, without suffering any disadvantage or losing any benefits they would have been entitled to if they had not withdrawn their consent. Patients will withdraw from the study (survival follow-up) in the following cases: ● Completion of the follow-up period, ● Withdrawal of consent for follow-up, ● Patients who could no longer be followed up on, ● Death, ● Termination of the study by the clinical trial sponsor, Institutional Review Board (IRB) / Ethics Committee (EC), regulatory authority, or other government authority.

[0447] Definition of Dose-Limiting Toxicity (DLT)

[0448] Throughout the study, DLTs are defined by the occurrence of study drug-related adverse events (Ae) (potentially or clearly related to the procedure) with CLN-619 administered alone, in combination with pembrolizumab, or in combination with chemotherapy. DLTs are defined separately for hematological, non-hepatic, and non-hematological events outlined below. The severity of all events is graded according to CTCAE v5.0. For patients experiencing a Grade 3 or higher adverse event, the procedure using the study drug(s) should generally be discontinued until the event is managed and resolved, and the relationship between the event and the administration of the study drug(s) should be evaluated.

[0449] Hematological dose-limiting toxicity (DLT): ● Grade 4 neutropenia lasting more than 7 days, ● Grade 4 febrile neutropenia, ● Grade 3 febrile neutropenia lasting more than 48 hours or accompanied by objective evidence of clinical infection. ● Grade 4 thrombocytopenia or grade 3 or higher thrombocytopenia with clinically significant bleeding.

[0450] Non-hepatic, non-hematological dose-limiting toxicity (DLT):

[0451] Non-hematological non-hepatic AEs of grade 3 or higher are considered DLTs, with the following exceptions: ● In other cases, grade 3 or 4 electrolyte abnormalities without clinical complications that respond to medical intervention within 96 hours. ● Grade 3 fatigue lasting less than 72 hours, ● Grade 3 overheating, ● Grade 3 headaches that respond to medical intervention within 24 hours. ● Grade 3 nausea, vomiting, or diarrhea that responds to medical intervention within 72 hours. ● Grade 3 or 4 elevated amylase or lipase in the absence of signs on X-ray or clinical signs and symptoms suggestive of pancreatitis. ● Grade 3 hypertension without clinical complications and responding to medical intervention within 24 hours.

[0452] Hepatic non-hematological dose-limiting toxicity (DLT):

[0453] Transaminase levels elevated ● Regardless of duration, an increase in AST or ALT of grade 3 or higher exceeding 10×ULN, ● Grade 3 elevated AST or ALT levels that do not improve to Grade 2 or lower within 7 days and to Grade 1 or lower within 14 days, and are between 5x ULN and 10x ULN.

[0454] Bilirubin levels rise ● Regardless of duration, an increase in total bilirubin of grade 3 or higher exceeding 5×ULN, ● Grade 3 bilirubin elevation of 3×ULN or higher and 5×ULN or lower, which does not improve to Grade 2 or lower within 7 days and to Grade 1 or lower within 14 days.

[0455] Adverse events that meet the criteria of Hy's Law are also considered DLTs: ● AST or ALT of 3×ULN or higher, ● Total bilirubin levels of 2×ULN or higher in the absence of cholestasis, ● Absence of verifiable alternative etiologies.

[0456] A persistent and clinically significant Grade 2 event that justifies ongoing medication discontinuation in the patient may also be considered a DLT on an individual basis.

[0457] Research discontinuation rules

[0458] If one Grade 5 event or two Grade 4 events that may be related to CLN-619 therapy are observed, patient enrollment in the study will be temporarily suspended. Grade 4 laboratory abnormalities that improve to Grade 2 or lower within 96 hours are not included.

[0459] Rules for determining the Module A accelerated titration cohort

[0460] Accelerated titration should be switched to a standard 3+3 design at a dose level of 3.0 mg / kg unless initiation at a dose level lower than 3.0 mg / kg is required due to toxicity at dose levels lower than 3.0 mg / kg.

[0461] At the completion of the single-patient accelerated titration cohort: ● If no Grade 2 or higher Ae associated with CLN-619 occurs during the first cycle, dose escalation can proceed to the next accelerated titration cohort. ● If a patient experiences any grade 2 or higher Ae associated with CLN-619 during the first cycle, the current cohort will be modified to a 3+3 design, enrolling an additional 2-5 patients. ● If the patient has experienced DLT, change the design to a 3+3 configuration and enroll an additional 2-5 patients.

[0462] Determination rules for the 3+3 cohorts with Module A dose escalation

[0463] At the completion of the 3+3 cohort (minimum 3 patients, maximum 6 patients): ● If none of the three patients have experienced DLT, the dose escalation can proceed to the next cohort. ● If one out of three patients has experienced DLT, enroll an additional three patients in the cohort. ● If one or fewer of the six patients have experienced a dose-limiting trial (DLT), the patient can proceed to the next cohort for dose escalation. ● If two or more patients have experienced a DLT, the MTD has been exceeded, and further enrollment at that dose level will be discontinued. ○ Next, cohorts with lower doses, including intermediate doses, may be further explored until the MTD is determined.

[0464] Determination rules for the Module B dose escalation 3+3 cohort

[0465] At the completion of the dose-escalation level cohort (minimum 3 patients, maximum 6 patients): ● If none of the three patients have experienced a dose-limiting trial (DLT), dose escalation can proceed to the next dose-level cohort. ● If one out of three patients has experienced a DLT, enroll an additional three patients in the current dose-level cohort. ● If one or fewer of the six patients have experienced DLT, the dose escalation can proceed to the next dose level cohort, provided that the safety of the next dose escalation level of CLN-619 has been established by the SRC during the Module A monotherapy dose escalation group in the study. ● If two or more patients have experienced a DLT, the MTD has been exceeded, and further enrollment at that dose level will be discontinued. ○ A "minus 1" cohort with a lower dose may be enrolled according to the 3+3 decision rule described above. - If two or more patients in the "minus 1" cohort have experienced DLT, the sponsor may continue with a lower-dose "minus 2" cohort according to the 3+3 decision rule above. • If there are two or more patients who have experienced DLT in the "minus 2" cohort, the sponsor will not initiate expansion of the Module B cohort.

[0466] Rules for determining the Module C dose escalation (3 mg / kg and 10 mg / kg)

[0467] To establish safety at a dose level of 3 mg / kg, ● If none of the three patients experience a dose level of 3 mg / kg, dose escalation can proceed to a dose level of 10 mg / kg. ● If one out of three patients has experienced a dose level of 3 mg / kg, enroll an additional three patients. ● If one or fewer of the six patients have experienced DLT, dose escalation can proceed to a dose level of 10 mg / kg. ● If two or more patients have experienced a dose level of 3 mg / kg, the median dose (MTD) has been exceeded, and further enrollment at 3 mg / kg will be discontinued.

[0468] To establish safety at a dose level of 10 mg / kg, ● If 0 out of 3 patients have experienced a dose-limiting trial (DLT) at a dose level of 10 mg / kg, then a dose increase at 10 mg / kg is permitted. ● If one out of three patients has experienced a dose level of 10 mg / kg, enroll an additional three patients at 10 mg / kg. ● If one or fewer out of six patients have experienced a dose-limiting trial (DLT) at a 10 mg dose level, the dose can be increased at 10 mg / kg. ● If two or more patients have experienced a dose-limiting trial (DLT) at 10 mg / kg, the MTD has been exceeded, and further enrollment at 10 mg / kg will be discontinued. Data from each dose level tested in combination with chemotherapy for each tumor type will be reviewed to determine the dose for enrolling the remaining patients in dose expansion (up to a total of 40 patients).

[0469] Rules for determining the dose escalation of Module D ● To evaluate the safety of the medication schedule (10 mg / kg, first cycle, Q1W and second cycle onward, Q3W), initially enroll 3 to 6 patients. If fewer than 2 dose-limiting toxicities (DLTs) are observed, additional patients (up to 10) may be enrolled to collect additional PK and biomarker data to establish this medication schedule. Additional patients may be enrolled if 0 out of 3 patients have experienced DLTs. ● If one out of three patients has experienced DLT, register an additional three patients. ● If one or fewer of the six patients have experienced DLT, additional patients can be enrolled. If two or more patients have experienced DLT, the MTD has been exceeded, and further enrollment under this medication schedule will be stopped.

[0470] Dosage Expansion Decision Rules

[0471] If, during the study, more than 33% of patients experience an Ae meeting the criteria for DLT during the first three-week cycle, or if safety findings suggesting clinically significant cumulative toxicity are observed, the SRC may be convened again to review data from the expanded cohort and develop recommendations for medication for continuing patients.

[0472] Determination of the maximum biologically effective dose

[0473] Both Module A (monotherapy dose escalation) and Module B (combination therapy dose escalation) are designed to identify the MTD, MAD (if MTD is not defined), and / or MBED for CLN-619 or CLN-619 in combination with pembrolizumab, respectively, and to inform the selection of RP2D for further investigation in each cohort expansion. During this phase, PK and exploratory biomarker data, in addition to safety and efficacy data, will be collected and analyzed to evaluate whether doses below the MTD or MAD may have a more favorable clinical profile.

[0474] Dosage timing and frequency

[0475] All infusions are performed by trained personnel in a monitoring environment with appropriate equipment and immediate access to medical care to manage potentially serious or life-threatening reactions. In dose escalation (modules A and B), CLN-619 is administered as a 60-minute IV infusion (+10 / -5 min) on day 1 of each 3-week treatment cycle. The dosage of CLN-619 is calculated using baseline body weight, which is the weight obtained at screening. The dosage should be adjusted if there is a + / - 10% change in body weight.

[0476] In Module B, pembrolizumab 200 mg is administered as a 30-minute IV infusion (+10 / -5 min) on day 1 of each 3-week treatment cycle. For further information regarding pembrolizumab administration, please refer to the Pharmacy Manual. When CLN-619 is administered in combination with pembrolizumab in Module B of this study, premedication containing dexamethasone is administered first, followed by pembrolizumab. CLN-619 should be administered immediately after completion of the pembrolizumab infusion (+30 minutes). If an infusion-related reaction occurs during pembrolizumab administration, the administration of the combination therapy may be delayed.

[0477] In Module C, standard therapeutic chemotherapy must be administered before CLN-619 administration on day 1 of each 3-week treatment cycle, according to prescribing information or institutional policy. In Module D, CLN-619 (10 mg / kg) is administered over 120 minutes on day 1 of cycle 1, which is the first of three weekly doses in cycle 1. For all subsequent weekly doses (C1D8 and C1D15) and cycles (from cycle 2 onward), infusions can be administered over 60 minutes if the patient has not experienced an IRR.

[0478] Prior to administering CLN-619, pre-infusion medication containing the required corticosteroids should be given. CLN-619 should be administered as a 120 (+10 / -5) minute IV infusion on day 1 of cycle 1. The CLN-619 dosage should be calculated using baseline body weight. If no infusion-related reaction occurs during the first cycle of treatment, 60 (+10 / -5) minute IV infusions should be used for all subsequent 3-week cycles. If an infusion-related reaction occurs during the visit on day 1 of cycle 1, a 120 (+10 / -5) minute IV infusion of CLN-619 should be used on day 1 of cycle 2, and 60 (+10 / -5) minute IV infusions should be used for subsequent cycles (Figures 26A and 26B). If an IRR occurs after the C2D1 visit, the clinical setting should contact the sponsor for instructions regarding subsequent infusions.

[0479] Premedication to prevent reactions associated with the injection

[0480] For patients treated with CLN-619 alone, in combination with pembrolizumab, or in combination with chemotherapy, premedication with corticosteroids, antihistamines, and / or antipyretics to prevent infusion-related reactions is essential. Suitable corticosteroids include dexamethasone, suitable antihistamines include diphenhydramine and famotidine, and suitable antipyretics include paracetamol, acetaminophen, and ibuprofen. Folic acid supplements may also be administered.

[0481] Grade classification and management of reactions associated with injection

[0482] Infusion reactions may occur with fever, chills, throbbing chills, headache, rash, itching, arthralgia, hypotension or hypertension, bronchospasm, or other symptoms.

[0483] Further recommended actions are shown in Table 8 below and may be modified as appropriate based on local treatment standards and guidelines. [Table 8-1] [Table 8-2] [Table 8-3]

[0484] Evaluation of disease response using tumor imaging and RECIST v1.1

[0485] Images are interpreted on-site by the principal investigator, and efficacy response evaluations are based on the principal investigator's assessment. Tumor imaging is very preferably obtained by contrast-enhanced CT. For the chest, abdomen, and pelvis, contrast-enhanced MRI may be used if iodine-enhanced CT is contraindicated or required by local practice. When brain imaging is clinically required, MRI is the very preferred modality for imaging the brain. If MRI is medically contraindicated, contrast-enhanced CT is an acceptable alternative.

[0486] To optimize the reproducibility of existing and new tumor volume assessments and improve the accuracy of imaging-based response or progression assessments, the same imaging techniques, ideally the same scanner, and the use of contrast agents should be applied to patients throughout the study.

[0487] Any patient who completes 34 cycles of treatment will undergo tumor imaging at their safety follow-up visit. Patients who discontinued treatment before 34 cycles do not need tumor imaging at this visit if it was performed within 12 weeks prior to the visit.

[0488] Disease evaluation using RECIST

[0489] RECIST v1.1 will be used as the primary measure for evaluating tumor response in this study and for making treatment decisions.

[0490] Confirmatory scans for patients showing an objective response should be performed at least four weeks after the initial scan in which the response was observed, or at the next scan required by the protocol.

[0491] Clinical stability is defined as follows: ● There are no symptoms or signs indicating clinically significant progression of the disease. ● No need for enhanced management, including increased pain relief, radiation therapy, or other palliative treatments. ● For further information on RECIST, please refer to https: / / recist.eortc.org / recist-1-1-2 / .

[0492] Pharmacokinetics

[0493] Blood samples are collected to evaluate serum concentrations (multiple concentrations possible) of CLN-619 and pembrolizumab (module B only), and related pharmacokinetic parameters are determined.

[0494] immunogenicity

[0495] Blood samples are collected and immunogenicity parameters, including human antidrug (CLN-619 and pembrolizumab (module B only)) antibody titers (ADA), are evaluated.

[0496] Definition of AE

[0497] An adverse event (AE) is any adverse medical event or worsening of a pre-existing medical condition in a clinical trial participant who has been administered a drug, and which is not necessarily causally related to the treatment.

[0498] Therefore, AE could be the following: ● Any undesirable and unintended clinical signs (including, for example, abnormal clinical laboratory findings). ● Any symptoms, illnesses, or injuries that are temporally related to the use of the investigational drug, whether or not they are related to the investigational drug.

[0499] Ae does not include the following: ● Medical or surgical procedures such as endoscopy, tooth extraction, or blood transfusion (however, the conditions that cause such procedures may be adverse events). ● A pre-existing disease or condition that is present at the start of the study and will not worsen during the study. ● Any situation in which no adverse medical events occurred (e.g., hospitalization for elective cosmetic surgery or social hospitalization). ● Clinical test results that the principal investigator determined to be clinically insignificant.

[0500] Adverse events may be spontaneously reported or elicited during open-ended questionnaires, examinations, or assessments with patients. (To prevent reporting bias, patients should not be asked about the specific occurrence of one or more adverse events.)

[0501] Chronic diseases that existed before study enrollment should be recorded in the medical history section of the eCRF, and should only be reported as Ae if an increase in the frequency or severity of the condition is observed during the study.

[0502] Spontaneous progression or worsening of malignant tumors during the study should be recorded as part of the efficacy evaluation and should not be recorded as an AE / SAE. Death due to disease progression should be recorded as part of the efficacy evaluation and should not be considered a SAE.

[0503] Signs and symptoms of disease progression should not be reported as Ae / SAE if they are clearly related to relapse or expected changes associated with disease progression of baseline malignancy. These signs and symptoms should only be reported as Ae / SAE (at the discretion of the investigator) if: ● If the principal investigator determines that the malignant tumor is abnormally severe or has accelerated progression, ● When the principal investigator believes that the signs and exacerbation of malignant tumors are directly caused by IMP.

[0504] If there is any uncertainty as to whether an AE is solely attributable to the malignant tumor under study, it should be reported as an AE / SAE as appropriate.

[0505] The principal investigator, who is a qualified physician, and any designated person are responsible for detecting, evaluating, documenting, and reporting events that meet the definition of an AE or SAE, and other reportable safety events. The principal investigator remains responsible for following up on Ae, SAE, and other reportable safety events for the sake of outcome.

[0506] Grade classification of the severity of adverse events

[0507] The severity of ae is recorded according to NCI CTCAE v5.0. This guide provides common terminology for describing severity levels, analyzing and interpreting data, scaling AE aggregate scores, and clarifying the clinical significance of all ae. A copy of the current version of NCI CTCAE is available for download from http: / / evs.nci.nih.gov / ftp1 / CTCAE / About.html.

[0508] For Ae grades not listed in NCI CTCAE, the following grading classification should be used: ● Grade 1: Mild AE - no symptoms or mild symptoms; clinical or diagnostic observation only; no intervention required. ● Grade 2: Moderate AE - requiring minimal, local, or non-invasive intervention; limitation of activities of daily living other than age-appropriate self-care. ● Grade 3: Severe AE - Severe or medically significant, but not immediately life-threatening; requires hospitalization or extended hospital stay; causes functional impairment; limits daily living activities. ● Grade 4: AEs that cause life-threatening or functional impairment - life-threatening consequences; require immediate intervention. ● Grade 5: Death related to AE

[0509] An AE (Accidental Exposure) classified as severe is not the same as a critical AE. Severity is a category used to assess the intensity of an event, and both Ae and SAE can be classified as severe. An AE is defined as "critical" if it meets one of the predefined critical outcomes listed below.

[0510] Sample size

[0511] Formal sample size estimation was not performed.

[0512] Module A:

[0513] The number of patients during the dose escalation phase was estimated to be up to 50 across all seven cohorts, including cohort extensions at all dose levels (45 patients were enrolled). It is estimated that up to 240 patients could be enrolled in the Module A cohort expansion (assuming full expansion). In total, it is estimated that up to approximately 285 patients will be administered in Module A of this trial.

[0514] During dose escalation, patients who discontinue treatment for reasons other than DLT during the 3-week DLT monitoring period will be replaced.

[0515] Module B:

[0516] The number of patients in the combined dose escalation is estimated to be up to 30 (assuming 6 patients per up to 5 dose levels), and 25 patients were enrolled. Up to 200 patients are estimated to be enrolled in the Module B cohort expansion. In total, it is estimated that up to approximately 225 patients will be treated in Module B of this trial.

[0517] Module C:

[0518] The number of patients in the chemotherapy combination cohort is a maximum of 40 patients per indication (40 each for NSCLC, endometrium, and epithelial ovary). In total, it is estimated that a maximum of approximately 120 patients will be dosed in Module C of this trial.

[0519] Module D:

[0520] In the Module D monotherapy loading dose cohort, a maximum of 10 patients will be enrolled.

[0521] During the dose escalation cohort, patients who withdraw from the study for reasons other than DLT during the 3-week DLT monitoring period will be replenished.

[0522] Results:

[0523] Sixty-four patients were administered CLN-619 in combination with pembrolizumab (n = 22) or CLN-619 as monotherapy (n = 42). Baseline characteristics are shown in Table 9 below.

[0524] [Table 9]

[0525] CPI, checkpoint inhibitor; NSCLC, non-small cell lung cancer.

[0526] a Other tumor types in the combination cohort: stomach (2 patients), esophagus (1), head and neck (1), skin (1), and urothelial carcinoma (1); b Other tumor types in the monotherapy cohort: breast (2), pancreas (2), sarcoma (2), adenoid cystic carcinoma (1), cecal cancer (1), duodenum (1), head and neck (1), kidney (1), leiomyosarcoma (1), mediastinal endometrioid sarcoma (1), melanoma (1), parotid gland (1), peritoneal mesothelioma (1), and thyroid (1).

[0527] Of the 22 patients treated with CLN-619 plus pembrolizumab, 18 were evaluable for response using RECIST. Four patients did not undergo post-baseline imaging for response evaluation due to withdrawal of consent (n=2), death due to disease progression (n=1), and transfer to hospice and acute kidney injury (n=1). Confirmed responses (all PRs) were observed in three patients treated with CLN-619 at doses of 3 mg / kg or higher in combination with pembrolizumab. Responses were observed in patients with tumor types that are not typically responsive to checkpoint inhibitors (CPIs) alone.

[0528] Table 10 details the characteristics of patients who responded to the treatment, and Figure 27 shows the duration of treatment and clinical activity in the combination therapy cohort.

[0529] [Table 10]

[0530] Efficacy in NSCLC was observed in both the monotherapy and combination therapy cohorts, as shown in Figure 28, which illustrates the duration of treatment and clinical activity in patients with NSCLC. Objective responses and stable disease (SD) were observed in patients with NSCLC with targetable oncogenic mutations in both the CLN-619 monotherapy and combination therapy cohorts.

[0531] Of the 42 patients treated with CLN-619 monotherapy, 29 received CLN-619 at doses of 1 mg / kg or higher and were evaluable by RECIST. Six patients did not undergo post-baseline imaging for response evaluation due to clinical progression (n=3), hospice care (n=1), and withdrawal of consent (n=2). Seven patients received sub-therapeutic doses (<1 mg / kg) or were not evaluable for response by RECIST. The clinical benefit rate (CBR) was 41.4% (1 complete response (CR), 2 partial responses (PR), and 9 stable disease (SD) for 18 weeks or longer). Table 11 shows the characteristics of patients with response or SD for 18 weeks or longer, and Figure 29 shows the duration of treatment and clinical activity in the 1 mg / kg or higher monotherapy cohort.

[0532] [Table 11]

[0533] Safety: Combination therapy with CLN-619 and pembrolizumab was well-tolerated at doses ranging from 1 to 6 mg / kg and demonstrated clinical activity, including objective responses, in tumor types typically refractory to pembrolizumab. No adverse events (AEs) meeting the DLT criteria defined in the protocol were observed, and most TEAEs were grade 1 / 2 (see Figure 30). The procedure-related AEs (TRAEs) reported in ≥10% of patients were fatigue (combination: 18.2%, monotherapy: 9.5%) and IRR (combination: 18.2%, monotherapy: 28.6%). The only grade 3 or higher procedure-related AE reported in ≥5% of patients in either group was increased AST (combination: 0%, monotherapy: 7.1%). TEAEs led to discontinuation of the study treatment in 13.6% (3 / 22 patients) of the combination therapy cohort and 9.5% (4 / 42 patients) of the monotherapy cohort. IRRs are known AEs that occur in CLN-619, and IRRs were the most frequently reported TRAE in CLN-619. When prophylactic premedication was administered, most IRRs were grade 1 or 2, occurred on day 1 of the first cycle, and resolved rapidly. No treatment-related deaths were reported.

[0534] Conclusion: Objective responses were observed with CLN-619 plus pembrolizumab in patients with tumor types typically refractory to pembrolizumab (e.g., NSCLC with ALKr and EGFRm). CLN-619 plus pembrolizumab was well-tolerated at doses ranging from 1 to 10 mg / kg. Longer-term follow-up of patients treated with CLN-619 monotherapy will confirm favorable safety and sustained clinical benefit, including objective responses in patients with multiple tumor types and disease progression after CPI therapy. Based on these findings, the expanded cohort enrolls patients with endometrial cancer and NSCLC in monotherapy and combination therapy cohorts. The best confirmed responses in RECIST-evaluable patients are shown in Table 12.

[0535] [Table 12]

[0536] ALKr: Anaplastic lymphoma kinase gene rearrangement; CBR: Clinical benefit rate; CR: Complete response; EGFRm: Epidermal growth factor receptor mutation; PR: Partial response; ORR: Objective response rate; SD: Disease stabilization.

[0537] CLN-619 also demonstrated monotherapy activity, including objective responses across multiple tumors.

[0538] Example 4. Conclusions from the use of CLN-619 (anti-MICA / B antibody) in solid tumors and rationale for the treatment of multiple myeloma.

[0539] CLN-619 will also be studied in a Phase 1 clinical trial (NCT06381141) in patients with relapsed / refractory multiple myeloma.

[0540] CLN-619 demonstrates antitumor efficacy in the treatment of advanced and solid tumors and is well-tolerated as monotherapy and in combination therapy.

[0541] In previous examples, the inventors demonstrated the clinical development of CLN-619, a humanized IgG1 monoclonal antibody that specifically binds to NKG2D ligands MICA and MICB, in patients with progressive solid tumors. These studies showed in vitro binding data that, overall, supports the highly specific and potent binding of CLN-619 to MICA / MICB, demonstrating broad reactivity and high affinity binding to representative allele variants of MICA and standard MICB allele variants. CLN-619 has multiple mechanisms of action. CLN-619 binds to MICA / MICB, preventing the proteolytic release of MICA / MICB from the surface of tumor cells and amplifying NKG2D-mediated immune cell activation accompanied by antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent phagocytosis (ADCP) in MICA / MICB-expressing tumor cells. Furthermore, in vivo studies demonstrated that CLN-619 treatment produced potent antitumor activity in mouse models with MICA / MICB-expressing human liver, lung, and multiple myeloma tumor xenografts across a dose range of 0.1 mg / kg to 10 mg / kg. Antitumor efficacy was also observed in a lung cancer model at a low dose of 0.1 mg / kg.

[0542] NCT05117476, described in the above example, enrolled patients with advanced and metastatic solid tumors in the CLN-619 study (CLN-619-001). Multiple doses of CLN-619 (maximum dose of 10 mg / kg) were tested administered every three weeks, either alone or in combination with pembrolizumab. In the study, CLN-619 was well-tolerated in both monotherapy and combination therapy settings, no dose-limiting toxicities were observed during dose escalation, and no grade 3 procedure-related adverse events (TRAEs) were observed in patients who received premedication as required by the protocol.

[0543] As of June 22, 2023, 61 patients had been exposed to intravenous doses of CLN-619 at 0.1, 0.3, 1, 3, 6, and 10 mg / kg, each on a 21-day cycle schedule. 43 patients were exposed to CLN-619 monotherapy, with 10 patients receiving a 3 mg / kg dose, 11 patients receiving a 6 mg / kg dose, and 12 patients receiving a 10 mg / kg dose. 18 patients were exposed to CLN-619 at 1, 3, 6, and 10 mg / kg in combination with pembrolizumab, with 6 patients receiving a 10 mg / kg dose. CLN-619 was found to be well-tolerated without adverse events (AEs) meeting the protocol-defined DLT criteria or grade 4 TRAEs.

[0544] Serious adverse events (SAEs) occurring under treatment were monitored after administration of CLN-619 as monotherapy and in combination with pembrolizumab. Only one infusion-associated reaction (IRR) event with laryngeal edema was observed, experienced in a patient at a dose level of 10.0 mg / kg, and was considered to be related to CLN-619. This patient had not received corticosteroid premedication. One non-serious grade 3 rash was reported in the monotherapy group and was considered potentially related. No serious adverse reactions to CLN-619 were reported in combination with pembrolizumab, but one SAE of acute kidney injury (AKI) was reported, which was not related to CLN-619. In patients treated with CLN-619 monotherapy, the most common treatment-induced adverse events (TEAEs) observed to date in more than 5% of patients were IRR (23.3%), abdominal pain (18.6%), fatigue (18.6%), and fever (18.6%).

[0545] Most IRRs occurred during or immediately after the first dose of CLN-619, one case occurred after the sixth dose, and one patient experienced IRRs both after the first and second doses. One event occurred in a patient receiving concomitant treatment with pembrolizumab. All patients recovered without sequelae. The treatment regimen recommends IRR premedication and a longer infusion time for the first dose of CLN-619. Monitoring for immune-related adverse events such as immune-mediated colitis, hepatitis, interstitial pneumonia, hypothyroidism, hyperthyroidism, adrenal insufficiency, hypophysitis, nephritis with renal insufficiency, myocarditis, and uveitis is also recommended.

[0546] Rationale for the use of CLN-619 in the treatment of multiple myeloma

[0547] Multiple myeloma (MM) is a plasma cell neoplasm resulting from the malignant transformation of antibody-producing plasma cells in the bone marrow, characterized by the presence of escalating levels of abnormal monoclonal immunoglobulin (M protein) in serum and urine. Overproduction of M protein can lead to fatal complications, including bone destruction, hypercalcemia, anemia, renal damage, and an increased risk of infection.

[0548] Multiple myeloma (MM) accounts for approximately 10% to 18% of hematological malignancies. It is a disease of the elderly, with a median age at diagnosis of approximately 70 years, and is slightly more common in men than in women. Multiple myeloma is 2 to 3 times more common in African American patients than in Caucasians, making it the most common hematological malignancy in this racial group. The global incidence of MM is approximately 160,000 cases per year. In the United States, there are approximately 34,500 new cases of MM each year, and about 13,000 deaths from MM. The frequency of MM is geographically heterogeneous, with the highest incidence in Europe, North America, and Australia / New Zealand. Despite recent advances, multiple myeloma remains incurable, and the most common causes of death are disease progression, infection, and renal failure. The median overall survival (OS) for patients with MM ranges from 2 to over 10 years, and approximately 15% of patients die within 2 years of diagnosis.

[0549] Treatment options for multiple myeloma (MM) include a variety of therapies, all of which work to control and eliminate multiple myeloma cells. These drugs include proteosome inhibitors, immunomodulators, steroids, antibodies, BCMA-targeted therapies, and chemotherapy. Unfortunately, most MM patients experience repeated relapses and require multiple lines of therapy. Relapsed MM cells typically exhibit a more malignant phenotype, resulting in shorter response durations and reduced survival rates. Therapy at this stage typically requires a multifaceted approach to address the diverse biological aspects of the disease and the full clinical spectrum of disease-related complications. Long-term disease control can best be achieved by combining therapies with different mechanisms of action, particularly those targeting key components of the immune system. Therefore, there is a significant need for new therapies for MM.

[0550] It is well established that MM cells can evade tumor cell surveillance by shedding MICA / MICB from their surface via proteolytic shedding. The loss of MICA / MICB from the cell surface allows tumor cells to evade recognition and destruction by immune cells, which is accompanied by immune cell dysregulation due to the internalization of NKG2D on the immune cell surface and impaired function and proliferation of NK and CD8+ T cells. High concentrations of shedding MICA (sMICA) have been observed across multiple tumor types in patient-derived serum and have been shown to correlate with shorter survival, for example, in MM where it is an independent prognostic factor for OS and PFS. MM patients with known sMICA presence show reduced levels of NKG2D in NK and CD8+ T cells, as well as decreased cytotoxicity of NK cells. Furthermore, shedding MICA (sMICB) has been shown to contribute to an immunosuppressive microenvironment through the polarization of macrophages to the M2 phenotype and the increase of myeloid suppressor cells (MDSCs) in the tumor microenvironment (TME). Preclinical data demonstrated the role of the MICA / MICB-NKG2D axis in immune surveillance by showing that overexpression of MICA in a human tumor model resulted in delayed tumor growth and increased survival in mice. In addition, NKG2D-deficient mice, while developmentally normal, exhibited impaired tumor surveillance in a model of spontaneously occurring malignant tumors.

[0551] Immunomodulatory agents such as lenalidomide and pomalidomide can enhance the expression of MICA and CD155 in both MM cell lines and primary MM cells through cereblon binding and downregulation of transcription factors IKZF1, IKZF3, and IRF4, thereby inducing degranulation of interferon-gamma from NK cells. Therefore, upregulated MICA / MICB in myeloma cells can be stabilized by the use of anti-MICA / MICB stabilizing antibodies, thus enhancing the elimination of myeloma cells by NKG2D-expressing NK and T cells. In addition, immunomodulatory agents such as lenalidomide have been reported to enhance NK cell activity by lowering the threshold required for NK cell activation by CD16A and other receptors. In a phase 2 study of lenalidomide-refractory patients, pomalidomide treatment resulted in functional activation of both circulating and bone marrow-infiltrating NK and T cells. These changes included increased NK and T cell numbers accompanied by upregulation of activating receptors, including NKG2D. In this embodiment, the inventors suggest that increased expression of MICA and MICB in malignant plasma cells, as well as improved immune cell activation through the combined use of CLN-619 and an immunomodulator, may further synergistically enhance these effects.

[0552] Therapies that effectively restore MICA / MICB expression on the surface of tumor cells have been proposed as an attractive approach for treating cancer patients with multiple myeloma, as they reverse tumor-mediated immunosuppression.

[0553] Immunotherapy has led to improvements in overall survival and quality of life, substantially altering the treatment landscape for patients with MM; however, the majority of patients with progressive disease still experience disease progression during or after multiple double or triple therapy combinations, highlighting an unmet need for patients with refractory disease. Strong rationale exists for modulating both innate and adaptive immune responses using CLN-619 monotherapy or in combination with pomalidomide + dexamethasone. This embodiment provides a planned combination regimen using FDA-approved MM standard treatments, supporting robust scientific rationale and clinical data for the combination of CLN-619 for MM disease. Apoptotic activity induced by pomalidomide + dexamethasone can induce cellular stress, which in turn can increase the expression of NKG2D ligands in MM cells. Further stabilization of these MICA and MICB ligands by CLN-619 can lead to improved recognition by immune cells and enhance ADCC, a key cytolytic mechanism mediated by NK cells. The combination of IMiD and a monoclonal antibody with Fc functionality similar to CLN-619 has been shown to have a synergistic effect in enhancing NK cell activity in a clinical setting. Furthermore, higher levels of sMICA are correlated with reduced overall survival in MM patients compared to sMICA-negative MM patients. sMICA has been shown to have immunosuppressive effects in the tumor microenvironment; therefore, the ability of CLN-619 to bind to sMICA / sMICB may help alleviate NK and T cell dysfunction, potentially leading to improved anti-myeloma activity.

[0554] Therefore, due to their respective mechanisms of action, this embodiment suggests that treatment of multiple myeloma with CLN-619 alone or in combination with pomalidomide + dexamethasone may enhance the antimyeloma effects of innate and adaptive immunity. Preferably, the method envisioned by this embodiment provides an effective treatment for relapsed / refractory multiple myeloma (R / R MM).

[0555] This example details an open-label, dose-finding study designed to address the safety, tolerability, and efficacy of CLN-619 monotherapy and CLN-619 in combination with pomalidomide and dexamethasone for the treatment of R / R MM.

[0556] Part 1A of the study will use a two-stage dose-escalation design during a 3-week DLT observation period (Cycle 1) for CLN-619 monotherapy and a 3-week DLT observation period (Cycle 4 or earlier) for CLN-619 combination therapy in Part 1C. An implementation of the 3+3 cohort design in Part 1 will be used to identify the optimal dose of CLN-619. After evaluating preliminary efficacy with CLN-619 monotherapy in Part 1A, patients will proceed to dose extension of monotherapy (Part 1B) or combination therapy (Part 1C). Clinical response data from the first four or fewer cycles of CLN-619 monotherapy in Part 1A will be used to determine a patient's participation in Part 1B or 1C. Patients with a very good partial response (VGPR) or better based on the International Myeloma Working Group (IMWG) response criteria will continue with their assigned dose of CLN-619 monotherapy (Part 1B). Patients with disease progression (PD), partial response (PR), minimal response (MR), or stable disease (SD) based on the IMWG response criteria should continue receiving their assigned dose of CLN-619 in combination with pomalidomide and dexamethasone (Part 1C).

[0557] Based on the overall data from Part 1 reviewed by the SRC, a cohort of CLN-619 patients receiving pomalidomide plus dexamethasone in combination from the first cycle will be initiated in Part 2.

[0558] Rational basis for dosage in the treatment of R / R multiple myeloma

[0559] Based on the overall safety, PK, and biomarker (sMICA) data from the FIH study (CLN-619-001, outlined in Examples 2 and 3) in patients with advanced and metastatic solid tumors, as well as the relative abundance of the target in MM patients, the starting dose of 3 mg / kg Q3W for CLN-619 was selected for this study.

[0560] A study of patients with locally advanced or metastatic solid tumors enrolled in the monotherapy group of Study CLN-619-001 and exposed to at least one dose of CLN-619 monotherapy provides a rationale for drug administration in MM. The aforementioned study explored the safety, efficacy, and pharmacokinetics of CLN-619 in patients with advanced solid tumors whose disease progressed after available therapies, at doses ranging from 0.1 mg / kg to 10 mg / kg. No DLTs were observed in any of the patients. Only one patient experienced a TRAE (grade 2 IRR associated with the initial infusion of CLN-619), but no serious TRAEs were observed in this drug cohort.

[0561] Early observations supporting the efficacy of CLN-619 monotherapy in advanced or metastatic solid tumors (i.e., the observations in Examples 2 and 3 described herein) were promising. Confirmed objective responses (RECIST v1.1) were observed at dose levels of 3 mg / kg or higher, namely one complete response (CR) in a patient with parotid gland cancer treated at a dose level of 3 mg / kg, and two partial responses (PR) in patients with endometrial cancer treated at dose levels of 3 and 10 mg / kg. Disease stabilization lasting at least six cycles was also observed at dose levels of 1 mg / kg or higher.

[0562] The preliminary PK and sMICA pharmacodynamics of CLN-619 were also determined in study CLN-619-001 (i.e., the studies in Examples 2 and 3 described herein). At steady state (cycle 3), exposure increased beyond dose-proportional exposure from 0.1 mg / kg to 1 mg / kg, but was nearly dose-proportional from 1 mg / kg to 10 mg / kg. The dose-cohort mean half-lives of CLN-619 ranged from 60.5 to 464 hours (2.5 to 19 days), and generally, longer half-lives were observed at higher doses, accompanied by dose-dependent systemic clearance decreases, likely resulting from saturation of target-mediated clearance (TMDD).

[0563] A dose-dependent increase in the fold change of serum sMICA concentration was observed after treatment with CLN-619. Individual sMICA profiles generally show a steady increase in serum concentration, particularly with continued treatment with higher dose levels of CLN-619. Binding of CLN-619 to sMICA likely leads to an increase in total sMICA serum concentration and an overall decrease in free sMICA. Since sMICA has immunosuppressive effects in the tumor microenvironment, reducing the concentration of free sMICA may contribute to the clinical activity of CLN-619. The saturation of TMDD and the pharmacodynamic response of sMICA indicate that near-complete target saturation is achieved at dose levels starting from 3 mg / kg.

[0564] MICA / MICB expression is often elevated in both solid tumors and multiple myeloma. Serum sMICA concentrations also tend to be elevated in both solid tumor and multiple myeloma patients, with higher concentrations associated with poorer prognosis. Some studies have reported that serum sMICA concentrations are higher in multiple myeloma patients compared to solid tumor patients, which may reflect increased shedding rates and / or higher overall tumor volume. Therefore, a dose of at least 3 mg / kg is recommended to achieve targeted saturation in MM patients.

[0565] In summary, CLN-619 was well-tolerated in patients with solid tumors up to 10 mg / kg, and objective responses were observed only at doses of 3 mg / kg or higher. Evidence of target saturation (both TMDD saturation and pharmacodynamics of total sMICA in serum) is thought to occur at approximately 3 mg / kg or higher in most patients. Therefore, the selected starting dose of 3 mg / kg in the Q3W dosing schedule for CLN-619 is expected to be a safe and effective target binding in patients with multiple myeloma. See Figure 31 for the overall study design.

[0566] Treatment of R / R multiple myeloma using CLN-619 and IMiD in combination.

[0567] Pomalidomide, marketed in the United States as Pomalyst®, is a thalidomide analog or immunomodulatory (IMiD) manufactured by Bristol Myers Squibb for the treatment of myelopathy (MM). When used in combination with dexamethasone, pomalidomide acts by inducing tumor cell apoptosis, enhancing T-cell and NK-cell-mediated immunity, and inhibiting pro-inflammatory cytokines.

[0568] The standard therapeutic dose of 4 mg was used in this study, and dose reduction is permitted to mitigate the toxicity observed in neutropenia and pneumonia in other studies.

[0569] Figure 31 shows the overall study design for the treatment of R / R MM. Since CLN-619 follows a 21-day dosing schedule, the inventors designed a shorter dosing schedule for pomalidomide to match the CLN-619 dosing schedule. The standard treatment dosing schedule for pomalidomide is typically 28 days, with 21 days of dosing and 1 week of rest. The current study design administers pomalidomide on days 1–14 and rests on days 15–21 in each 21-day treatment cycle.

[0570] Patients should be treated with pomalidomide in combination with dexamethasone and CLN-619 until disease progression, unacceptable toxicity, or up to two years first occur.

[0571] Dexamethasone is a corticosteroid used to treat various diseases by suppressing the immune system. Dexamethasone is typically used in combination with other drugs for the treatment of multiple myeloma.

[0572] Various doses of dexamethasone, such as 20 mg (75 years and older) or 40 mg (under 75 years) in weekly doses, which have shown efficacy in treating R / R MM, were used in combination with pomalidomide.

[0573] In combination with pomalidomide and CLN-619, the current study design involves administering 20 mg of dexamethasone orally or intravenously on days 1, 8, and 15 of a 21-day treatment cycle, regardless of age.

[0574] Patients should be treated with dexamethasone in combination with CLN-619 and pomalidomide until disease progression, unacceptable toxicity, or up to two years, whichever comes first.

[0575] In the treatment cycle, patients who meet all eligibility criteria will complete the pre-medication evaluation upon their first day visit. After completing the pre-medication evaluation on day 1, the first dose of the study treatment (CLN-619) will be administered under observation at the facility, followed by a post-medication evaluation. The study treatment will consist of intravenous infusion of CLN-619, depending on the part and cohort.

[0576] In Part 1A (Dose Escalation), the treatment consists of the first four cycles of CLN-619 monotherapy. A cohort of patients with R / R MM is treated with escalating doses of CLN-619 monotherapy using a standard 3+3 dose escalation design. CLN-619 is administered at 3 and 6 mg / kg in 21-day cycles. Prophylactic medication to reduce infusion-related reactions is administered on the first day of the treatment and on day 1 of subsequent cycles.

[0577] The decision-making rules guiding dose escalation decisions and study implementation are based on DLTs or MADs occurring in patients during the initial three weeks of CLN-619 monotherapy treatment. Any DLTs occurring in subsequent cycles will be considered in determining the recommended optimal dose.

[0578] A 3+3 design will be used to assess toxicity in each cohort. At the completion of the 3+3 cohort (minimum 3 patients, maximum 6 patients): ● If 0 out of 3 patients have experienced DLT, dose escalation can proceed to the next cohort. ● If one out of three patients has experienced DLT, enroll an additional three patients in the cohort. ● If two or more patients have experienced DLT, the MTD is exceeded, and further enrollment in that cohort will be stopped. ○ Next, cohorts with lower doses, including intermediate doses, may be further explored until the MTD is determined.

[0579] Patients who are unable to complete the DLT observation period for reasons other than DLT experience will be excluded when estimating parameters, but all patient data will be included in the safety summary and data list.

[0580] Parts 1B and 1C CLN-619 Dose Extension: Patients who demonstrate acceptable safety and tolerability upon completion of the first four cycles of CLN-619 monotherapy in Part 1A will proceed to Part 1B or 1C based on the efficacy evaluation outcome.

[0581] Part 1B CLN-619 monotherapy: Patients who achieve a VGPR or better response rate based on the IMWG response criteria after 4 cycles of treatment should continue with a specific monotherapy dose of CLN-619 from the 5th cycle onward.

[0582] Part 1C CLN-619 + Pomalidomide + Dexamethasone: Patients who have achieved PR, MR, or SD based on IMWG response criteria after 4 cycles of CLN-619 monotherapy will receive CLN-619 in combination with pomalidomide and dexamethasone starting from the 5th cycle. Patients who have PD based on IMWG criteria at any point in time at the completion of cycles 1-4 will receive CLN-619 in combination with pomalidomide and dexamethasone. A DLT observation period is established during the first cycle (days 1-21) of the combination therapy. CLN-619 is administered in 21-day cycles. Pomalidomide is administered orally at a dose of 4 mg per day from day 1 of the 4th cycle, through days 1-14 of the repeated 21-day cycles. Dexamethasone is administered orally or intravenously at a dose of 20 mg on days 1, 8, and 15 of each cycle. Patients in Part 1C should be carefully monitored for DLT during the first three-week cycle of concomitant treatment.

[0583] If, at any point in the study in Part 1B or 1C, more than 30% of patients experience an AE that meets the DLT criteria during or after the defined DLT observation period, or if safety findings suggest clinically significant cumulative toxicity, a Series Research Committee (SRC) may be convened to review data from the extended cohort and develop recommendations for continued medication for patients.

[0584] Part 2: Based on the available safety and efficacy data from Part 1, a single dose of CLN-619 in combination with pomalidomide + dexamethasone will be selected, and patients will be enrolled in Part 2 to receive CLN-619 in combination with pomalidomide + dexamethasone from the first cycle of treatment. CLN-619 will be administered in 21-day cycles. Pomalidomide will be administered orally at a dose of 4 mg per day on days 1–14 of each repeated 21-day cycle. Dexamethasone will be administered orally or intravenously at a dose of 20 mg on days 1, 8, and 15 of each cycle. If, at any point during the study in Part 2, more than 30% of patients experience an AE meeting the DLT criteria, or if safety findings suggest clinically significant cumulative toxicity, a SRC may be convened to review data from the extended cohort and develop recommendations for continued patient dosing.

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Claims

1. A method for treating endometrial cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (anti-MICA / B antibody).

2. The method according to claim 1, wherein the subject has previously been treated with a PD-1 inhibitor (e.g., an anti-PD-1 antibody).

3. The method according to claim 1, wherein the subject has previously been treated with a VEGF inhibitor or hormone therapy.

4. The method according to claim 1, wherein the subject has never been treated with a PD-1 inhibitor (e.g., an anti-PD-1 antibody) in the past.

5. The method according to any one of claims 1 to 4, wherein the endometrial cancer is HER2+.

6. The method according to any one of claims 1 to 4, wherein the endometrial cancer is ER+ or PR+, or both.

7. A method for treating parotid gland cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (anti-MICA / B antibody).

8. The method according to claim 7, wherein the parotid gland cancer overexpresses one or both of EGFR and HER2.

9. The method according to claim 7 or 8, wherein the parotid gland cancer is mucoepidermoid carcinoma of the parotid gland.

10. The method according to any one of the prior claims, wherein the cancer is one or more variants of EGFR, TP53, HER2, PTEN, ARID1A, CTNNB1, or PLK3R1.

11. A method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (anti-MICA / B antibody), wherein the subject has a history of cancer (i) VEGF inhibitors, (ii) EGFR inhibitors, and / or (iii) Hormone therapy The aforementioned method, which has been used for treatment.

12. The method according to claim 11, wherein the subject responds to the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy (for example, showing a complete or partial response), and optionally, the subject has a response period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

13. The method according to claim 12, wherein the subject progresses after the period of response to the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy.

14. The method according to claim 13, wherein the administration of the anti-MICA / B antibody is performed after the progression following the response period to the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy.

15. The method according to claim 11, wherein the subject shows disease stabilization upon administration of the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy, and optionally, the subject has a disease stabilization period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months, or longer.

16. The method according to claim 11, wherein the treatment using the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy is discontinued before the commencement of administration of the anti-MICA / B antibody.

17. The method according to claim 11, wherein the treatment using the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy is continued after the commencement of administration of the anti-MICA / B antibody.

18. The method according to claim 11, wherein the subject's condition progresses when treated with the VEGF inhibitor, the EGFR inhibitor, or the hormone therapy.

19. The method according to any one of the prior claims, wherein the subject shows a response (e.g., complete response or partial response) or stabilization of disease after administration of the anti-MICA / B antibody.

20. The method according to claim 19, wherein the response after administration of the anti-MICA / B antibody lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, or 60 months, or longer.

21. The method according to any one of claims 11 to 20, wherein the EGFR inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI) (e.g., gefitinib, erlotinib, afatinib, dacomitinib, or osimertinib) or an anti-EGFR antibody (e.g., cetuximab, panitumumab, or nesitumumab).

22. The method according to 11 to 20, wherein the VEGF inhibitor comprises a small molecule tyrosine kinase inhibitor (TKI) (e.g., sunitinib, sorafenib, axitinib, pazopanib, or lenvatinib) or an anti-VEGF antibody (e.g., bevacizumab or ranibizumab).

23. The method according to 11 to 20, wherein the hormone therapy comprises an aromatase inhibitor (AI) (e.g., anastrozole), a selective estrogen receptor modulator (SERM), a luteinizing hormone-releasing hormone (LHRH) agonist, an antiandrogen, a CYP17 inhibitor, a progestin, an anti-adrenergic agent, or an estrogen receptor antagonist.

24. A method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (anti-MICA / B antibody), wherein the subject has previously been treated with a PD-1 inhibitor (e.g., an anti-PD-1 antibody), (i) The subject responded to the PD-1 inhibitor (e.g., showed a complete response or a partial response), (ii) The subject showed disease stabilization when administered the PD-1 inhibitor. The aforementioned method.

25. The method according to claim 24, wherein the subject has a response period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months or longer.

26. The method according to claim 25, wherein the subject progresses after the period of response to the PD-1 inhibitor.

27. The method according to claim 26, wherein the administration of the anti-MICA / B antibody is performed after the progression of the disease following the response period to the PD-1 inhibitor.

28. The method according to claim 24, wherein the subject had a disease stabilization period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, 60 months or longer.

29. The method according to claim 24, wherein the treatment using the PD-1 inhibitor is discontinued before the administration of the anti-MICA / B antibody is initiated.

30. The method according to claim 24, wherein the treatment using the PD-1 inhibitor is continued after the commencement of administration of the anti-MICA / B antibody.

31. The method according to any one of the prior claims, wherein, after administration of the anti-MICA / B antibody, the subject shows a response (e.g., complete response or partial response) or stabilization of disease.

32. The method according to claim 37, wherein the response after administration of the anti-MICA / B antibody lasts for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 42, 48, 54, or 60 months, or longer.

33. The method according to any one of the prior claims, wherein the PD-1 inhibitor is an anti-PD-1 antibody.

34. The method according to any one of the prior claims, wherein the anti-PD-1 inhibitor comprises pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

35. The method according to any one of the prior claims, wherein the subject has previously been treated with a VEGF inhibitor, an EGFR inhibitor, and / or hormone therapy.

36. A method for treating cancer in a subject, (i) The subject shall receive an initial dose of an antibody that binds to MICA / B (anti-MICA / B antibody), (ii) The subject is given a therapeutically effective dose of corticosteroid before the first dose of the anti-MICA / B antibody. The method, including the method described above.

37. The method according to claim 36, wherein the corticosteroid is administered 30 to 60 minutes before the administration of the anti-MICA / B antibody.

38. The method according to claim 36 or 37, wherein the corticosteroid is administered in a dose of about 2 mg to about 50 mg.

39. The method according to claim 36 or 37, wherein the corticosteroid is administered in a dose of about 10 mg.

40. The method according to any one of claims 36 to 39, wherein the corticosteroid is administered orally or intravenously.

41. The method according to any one of claims 36 to 40, further comprising administering one or more subsequent doses of the anti-MICA / B antibody.

42. The method according to claim 41, wherein the corticosteroid is administered before the first dose of the anti-MICA / B antibody, but not before the one or more subsequent doses of the anti-MICA / B antibody.

43. The method according to any one of claims 36 to 42, wherein the corticosteroid is dexamethasone.

44. The method according to claim 44, wherein the cancer is a tumor of the female reproductive organs, a salivary gland tumor, breast cancer, prostate cancer, lung cancer, or colon cancer.

45. The aforementioned tumor of the female reproductive organs includes endometrial tumor, ovarian cancer, or cervical cancer. The aforementioned salivary gland tumors include mucoepidermal tumors such as parotid gland tumors. The method according to claim 44.

46. The method according to any one of the prior claims, wherein the subject is a cancer which is a parotid gland cancer, cervical cancer, endometrial cancer, breast cancer, colon cancer, ovarian cancer, prostate cancer, sarcoma, melanoma, salivary gland adenoid cystic neoplasm, peritoneal mesothelioma, squamous cell carcinoma of the rectum, leiomyosarcoma, colorectal cancer, renal cancer, thyroid cancer, NSCLC, duodenal cancer, pancreatic cancer, mediastinal endometrial sarcoma, head and neck cancer, or cecal cancer.

47. The aforementioned subjects are as follows: Cancer characterized by low levels of PD-L1 expression, Cancer characterized by high levels of PD-L1 expression, Cancers characterized by low tumor mutational load (TMB), Cancer characterized by high tumor mutational load (TMB), Immunologically cold cancer, A cancer that is hot from an immunological perspective, Hormone-sensitive cancer, Cancer characterized by overexpression of oncogenic drivers, or Cancers that express one or more of the following: EGFR, ER, PR, or HER2. The method according to any of the prior claims, wherein the cancer is characterized by one or more of the above.

48. The method according to any one of the prior claims, wherein the subject has metastatic cancer.

49. The method according to any one of the prior claims, wherein the subject has previously received therapy including chemotherapy.

50. The method according to any one of the prior claims, wherein the anti-MICA / B antibody is administered in a dose of approximately 3 mg / kg to approximately 10 mg / kg.

51. The method according to any one of the prior claims, wherein the anti-MICA / B antibody is administered to the subject according to an administration interval (e.g., a cycle).

52. The method according to any one of the prior claims, wherein the administration interval includes a three-week cycle, and the anti-MICA / B antibody is administered once every three weeks (Q3W).

53. The method according to claim 51 or 52, wherein the administration interval is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times.

54. The method according to claim 53, wherein the repetition of the administration interval is carried out over a period of at least two weeks, three weeks, four weeks, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, twelve months, thirteen months, fourteen months, fifteen months, sixteen months, seventeen months, eighteen months, nineteen months, twenty months, twenty-one months, twenty-two months, twenty-three months, twenty-four months, three years, four years, or five years.

55. The method according to any one of the prior claims, wherein the anti-MICA / B antibody comprises a light chain variable region (VL) including light chain complementarity determination region 1 (LCDR1) of SEQ ID NO: 1, light chain complementarity determination region (LCDR2) of SEQ ID NO: 2, light chain complementarity determination region 3 (LCDR3) of SEQ ID NO: 3, and a heavy chain variable region (VH) including heavy chain complementarity determination region 1 (HCDR1) of SEQ ID NO: 4, heavy chain complementarity determination region 2 (HCDR2) of SEQ ID NO: 5, and heavy chain complementarity determination region 3 (HCDR3) of SEQ ID NO:

6.

56. The method according to claim 55, wherein the VL includes the amino acid sequence of Sequence ID No. 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

57. The method according to claim 55 or 56, wherein the VH comprises the amino acid sequence of Sequence ID No. 8, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

58. The method according to any one of claims 55 to 57, wherein the light chain comprises the amino acid sequence of Sequence ID No. 9, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

59. The method according to any one of claims 55 to 58, wherein the heavy chain comprises the amino acid sequence of Sequence ID No. 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

60. The method according to any one of the prior claims, wherein the anti-MICA / B antibody specifically binds to the MICA protein, the MICB protein, or both the MICA protein and the MICB protein.

61. The method according to any one of the prior claims, wherein the anti-MICA / B antibody binds to the alpha-3 domain of the MICA protein, the MICB protein, or both the MICA protein and the MICB protein.

62. The method according to claim 60 or 61, wherein the MICA protein is a membrane-bound MICA protein, a soluble MICA protein, or both.

63. The method according to claim 60 or 61, wherein the MICB protein is a membrane-bound MICB protein, a soluble MICB protein, or both.

64. The method according to any one of the prior claims, wherein the anti-MICA / B antibody is selected from whole immunoglobulin, scFv, Fab, F(ab')2, or disulfide bonded Fv.

65. The method according to any one of the prior claims, wherein the anti-MICA / B antibody is IgG or IgM.

66. The method according to any one of the prior claims, wherein the anti-MICA / B antibody is a humanized or chimeric antibody.

67. The method according to any one of the prior claims, further comprising administering an effective amount of a PD-1 inhibitor to the subject.

68. The method according to claim 67, wherein the PD-1 inhibitor is an anti-PD-1 antibody.

69. The method according to claim 67 or 68, wherein the anti-PD-1 inhibitor comprises pimivarimab, pembrolizumab, nivolumab, semiprimab, AMP-224, APM-514, or spartalizumab.

70. A method for treating multiple myeloma in a subject, comprising administering to the subject a therapeutically effective amount of an antibody that binds to MICA / B (anti-MICA / B antibody).

71. The method according to claim 70, further comprising administering a therapeutically effective amount of an immunomodulator to the subject.

72. The method according to claim 71, further comprising administering a therapeutically effective amount of dexamethasone to the subject.

73. The method according to any one of claims 70 to 72, wherein, after administration of the anti-MICA / B antibody, the subject shows a response (e.g., complete response or partial response) or stabilization of disease.

74. The method according to any one of claims 70 to 73, wherein the subject has relapsed / refractory multiple myeloma.

75. The method according to any one of claims 70 to 74, wherein the subject is unresponsive to a corticosteroid, melphalan, or a combination of vincristine, doxorubicin, and dexamethasone.

76. The method according to claim 70, further comprising administering a therapeutically effective dose of corticosteroid to the subject before the first dose of the anti-MICA / B antibody.

77. The method according to claim 76, wherein the corticosteroid is administered 30 to 60 minutes before the administration of the anti-MICA / B antibody.

78. The method according to claim 76, wherein the corticosteroid is administered in a dose of about 2 mg to about 50 mg.

79. The method according to claims 76 to 78, wherein the corticosteroid is administered in a dose of about 10 mg.

80. The method according to any one of claims 76 to 79, wherein the corticosteroid is administered orally or intravenously.

81. The method according to any one of claims 76 to 80, further comprising administering one or more subsequent doses of the anti-MICA / B antibody.

82. The method according to claim 81, wherein the corticosteroid is administered before the first dose of the anti-MICA / B antibody, but not before the one or more subsequent doses of the anti-MICA / B antibody.

83. The method according to any one of claims 76 to 82, wherein the corticosteroid is dexamethasone.

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